JP2024518497A - Combination therapy - Google Patents

Abstract

Disclosed is a combination comprising a therapeutically effective amount of a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a therapeutically effective amount of a BCL-2 inhibitor, and, optionally, a therapeutically effective amount of at least one other anti-neoplastic agent. Also disclosed is a method of using such a combination to treat a subject diagnosed with a hematopoietic disorder. The compound is represented by the following formula (I):

Description

The present invention relates to a novel combination comprising a therapeutically effective amount of a menin mixed lineage leukemia 1 (menin-MLL) inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a therapeutically effective amount of a B-cell lymphoma 2 (BCL-2) inhibitor, and, optionally, a therapeutically effective amount of at least one other anti-neoplastic agent, and to a method for treating a subject diagnosed with a hematopoietic disorder.

Of the 10 million cancer deaths recorded by GLOBOCAN in 2020, 7.1% are attributable to hematopoietic disorders. Therefore, new therapies are urgently needed for hematopoietic disorders such as acute myeloid leukemia (AML), myelodysplastic syndromes (MDS) and acute lymphoblastic leukemia (ALL), as further detailed below.

AML is a common hematological malignancy whose incidence rises from 3:100,000 in young adults to over 20:100,000 in older adults. In patients under 60 years of age, overall survival (OS) is 40-50%, but only 5% in patients over 60 years of age. The majority of patients newly diagnosed with AML are over 60 years of age. In this patient population, standard induction chemotherapy is often not an option due to increased treatment-related mortality as a result of age and comorbidities. The standard of care for AML patients who are not suitable for combination chemotherapy is treatment with hypomethylating agents (azacitidine or decitabine) or low-dose cytarabine. Despite these state-of-the-art treatments, the median OS is only about 10 months. In all types of AML, disease relapse is common despite initial treatment response and is the most common reason for death. Standard chemotherapy and allogeneic stem cell transplantation (when used) often fail to eradicate all tumor-propagating cells and select for chemotherapy-resistant leukemic propagating subclones. Patients refractory to salvage therapy are treated palliatively as current treatment options are extremely limited. The median survival for these patients is 2 months. Furthermore, patients with newly diagnosed intermediate or higher risk MDS and those who relapse after standard care have poor prognosis and high risk of progression to AML. Thus, new therapies are urgently needed for relapsed/refractory (R/R) AML and MDS patients, newly diagnosed AML patients who are ineligible for induction chemotherapy based on age and comorbidities, and newly diagnosed intermediate/high/very high risk MDS patients.

ALL is a hematological malignancy propagated by impaired differentiation, proliferation, and accumulation of lymphoid progenitor cells in the bone marrow and/or extramedullary sites. ALL represents 12% of all leukemia cases and is the most common childhood acute leukemia, with a worldwide incidence estimated at 1-4.75 per 100,000. ALL represents approximately 20% of adult leukemias. Despite high complete remission (CR) rates (80%-90%) with current therapies, the majority of adult patients with ALL will relapse. Five-year overall survival is approximately 30-40% in adults and elderly patients.

Therefore, new therapies are urgently needed for relapsed/refractory ALL, especially in adults and especially in elderly patients.

An embodiment of the present invention relates to a novel combination of a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and, optionally, at least one other anti-neoplastic agent.

Embodiments of the present invention relate to the use of a menin-MLL inhibitor as described herein in combination with a BCL-2 inhibitor and, optionally, at least one other anti-neoplastic agent to treat a subject diagnosed with a hematopoietic disorder, such as, but not limited to, a hematologic cancer.

Embodiments of the present invention relate to novel methods for treating a subject diagnosed with a hematopoietic disorder using such combinations. An embodiment of the novel method comprises administering to the subject a therapeutically effective amount of a menin-MLL inhibitor as described herein, a therapeutically effective amount of a BCL-2 inhibitor, and, optionally, a therapeutically effective amount of at least one other anti-neoplastic agent, where the menin-MLL inhibitor is a compound of formula (I), or a pharma- ceutical acceptable salt or solvate thereof.

Embodiments of the present invention relate to novel methods for treating a subject diagnosed with a hematopoietic disorder using such combinations. An embodiment of the novel method comprises administering to the subject a therapeutically effective amount of a menin-MLL inhibitor as described herein, a therapeutically effective amount of a BCL-2 inhibitor, and a therapeutically effective amount of at least one other anti-neoplastic agent, where the menin-MLL inhibitor is a compound of formula (I), or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, the present invention relates to a method of treating a subject diagnosed with a hematopoietic disorder, comprising administering to the subject a therapeutically effective amount of a menin-MLL inhibitor of formula (I) or a pharma- ceutical acceptable salt or solvate thereof, a therapeutically effective amount of venetoclax or a pharma-ceutical acceptable salt or solvate thereof, and a therapeutically effective amount of azacitidine or a pharma-ceutical acceptable salt or solvate thereof.

In some embodiments, the present invention relates to a method of treating a subject diagnosed with a hematopoietic disorder, comprising administering to the subject a therapeutically effective amount of a menin-MLL inhibitor of formula (I) or a pharma- ceutical acceptable salt or solvate thereof, a therapeutically effective amount of venetoclax or a pharma-ceutical acceptable salt or solvate thereof, and a therapeutically effective amount of azacitidine or a pharma-ceutical acceptable salt or solvate thereof, wherein the venetoclax or a pharma-ceutical acceptable salt or solvate is administered to the subject prior to, concurrently with, or after administration of the menin-MLL inhibitor, and the azacitidine or a pharma-ceutical acceptable salt or solvate is administered to the subject prior to, concurrently with, or after administration of the menin-MLL inhibitor.

In an embodiment, the menin-MLL inhibitor of formula (I) is:

のもの、並びにその互変異体及び立体異性形態［式中、
Ｒ^１ａは、－Ｃ（＝Ｏ）－ＮＲ^ｘａＲ^ｘｂ、Ｈｅｔ、又は

and tautomeric and stereoisomeric forms thereof,
R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

を表し、
Ｈｅｔは、１、２又は３個の窒素原子及び任意選択的にカルボニル部分を含有する５又は６員単環式芳香環を表し、
当該５又は６員単環式芳香環は、任意選択的に、Ｃ_３～６シクロアルキル及びＣ_１～４アルキルからなる群から選択される１又は２個の置換基で置換され、
Ｒ^ｘａ及びＲ^ｘｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^１ｂは、Ｆ又はＣｌを表し、
Ｙ^１は、－ＣＲ^５ａＲ^５ｂ－、－Ｏ－、又は－ＮＲ^５ｃ－を表し、
Ｒ^２は、水素、ハロ、Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－ＮＲ^７ａＲ^７ｂからなる群から選択され、
Ｕは、Ｎ又はＣＨを表し、
ｎ１、ｎ２、ｎ３及びｎ４は、１及び２からそれぞれ独立して選択され、
Ｘ^１はＣＨを表し、Ｘ^２はＮを表し、
Ｒ^４は、イソプロピルを表し、
Ｒ^５ａ、Ｒ^５ｂ、Ｒ^５ｃ、Ｒ^７ａ及びＲ^７ｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^３は、－Ｃ_１～６アルキル－ＮＲ^８ａＲ^８ｂ、－Ｃ_１～６アルキル－Ｃ（＝Ｏ）－ＮＲ^９ａＲ^９ｂ、－Ｃ_１～６アルキル－ＯＨ、又は－Ｃ_１～６アルキル－ＮＲ^１１－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル－Ｏ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルを表し、ここで、Ｒ^３の定義におけるＣ_１～４アルキル又はＣ_１～６アルキル部分のそれぞれは、互いに独立して、シアノ、ハロ、－ＯＨ、及び－Ｏ－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されていてもよく、
Ｒ^８ａ及びＲ^８ｂは、水素、Ｃ_１～６アルキル、－Ｃ（＝Ｏ）－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１２ａＲ^１２ｂ、並びに、－ＯＨ、シアノ、ハロ、－Ｓ（＝Ｏ）_２－Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１０ａＲ^１０ｂ及び－ＮＲ^１０ｃ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されているＣ_１～６アルキル、からなる群からそれぞれ独立して選択され、
Ｒ^９ａ、Ｒ^９ｂ、Ｒ^１０ａ、Ｒ^１０ｂ、Ｒ^１０ｃ、Ｒ^１１、Ｒ^１２ａ、及びＲ^１２ｂは、水素及びＣ_１～６アルキルからなる群からそれぞれ独立して選択される］、
並びにそれらの医薬的に許容される塩及び溶媒和物である。

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety;
the 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl, wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ may be substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C _1-6 alkyl substituted with _1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) _{2 -C 1-4} alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O _{)-C 1-4} alkyl;
R ^9a , R ^9b , R ^10a , R ^10b , R ^10c , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide besylate (benzenesulfonate):

及びその溶媒和物である。

and solvates thereof.

Those of ordinary skill in the art will understand that "and solvates thereof" refers to the besylate salt of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide besylate or a hydrate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate or a solvate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate or a hydrate thereof.

In particular, the present invention relates to (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate 0.5 to 2.0 equivalent hydrate.

In particular, the present invention relates to (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate 2.0 equivalent hydrate.

In certain embodiments, the menin-MLL inhibitor of formula (I) is crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate.

In certain embodiments, the menin-MLL inhibitor of formula (I) is crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate 0.5-2.0 equivalent hydrate.

More specifically, the present invention relates to crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate 2.0 equivalent hydrate.

Additional embodiments, features, and advantages of the invention will become apparent from the following detailed description, as well as through practice of the invention.

FIG. 1 is an X-ray powder diffraction (XRPD) pattern of compound A4: (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bisbesylate hydrate crystalline form A. FIG. 2 shows a comparison of tumor volumes as a function of time for a control group and treatment groups treated with regimens containing various amounts of Compound A3. FIG. 3 shows a comparison of percent tumor survival as a function of time (eg, Kaplan-Meier survival curves) for a control group and treatment groups treated with regimens containing various amounts of Compound A3. FIG. 4A shows a comparison of percent survival as a function of time in mice bearing established OCI-AML3 tumors following treatment with vehicle, either venetoclax, azacitidine or compound A1 monotherapy, either dual combinations of venetoclax and azacitidine, or compound A1 and venetoclax, or the triple combination of compound A1, venetoclax and azacitidine. FIG. 4B shows a comparison of percent survival of mice bearing established MOLM-13 tumors following treatment with vehicle, either venetoclax, azacitidine or compound A1 monotherapy, either dual combinations of venetoclax and azacitidine or compound A1 and venetoclax, or the triple combination of compound A1, venetoclax and azacitidine as a function of time. FIG. 5A is a contour plot of maxR showing the effect of compound A3 in combination with venetoclax on the proliferation of MOLM-13 cells in vitro. FIG. 5B is a contour plot of maxR showing the effect of Compound A3 in combination with azacytidine and venetoclax on the proliferation of MOLM-13 cells in vitro. FIG. 6A is a contour plot of maxR showing the effect of Compound A4 in combination with decitabine on the proliferation of MOLM-13 cells in vitro. FIG. 6B is a contour plot of maxR showing the effect of Compound A4 in combination with decitabine and venetoclax on the proliferation of MOLM-13 cells in vitro. FIG. 7A is a contour plot of maxR showing the effect of Compound A4 in combination with decitabine on the proliferation of OCI-AML3 cells in vitro. FIG. 7B is a contour plot of maxR showing the effect of Compound A4 in combination with decitabine and venetoclax on the proliferation of OCI-AML3 cells in vitro.

As used herein, the term "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo.

As used herein, the prefix "C _xy " (where x and y are integers) refers to the number of carbon atoms in a given group, i.e., a C _1-6 alkyl group contains 1 to 6 carbon atoms, and so on.

The term "C _1-4 alkyl" as used herein as a group or part of a group represents a straight or branched chain saturated hydrocarbon radical having from 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, etc.

The term "C _1-6 alkyl" as used herein as a group or part of a group represents a straight or branched chain saturated hydrocarbon radical having from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, n-pentyl, n-hexyl, and the like.

The term "C _3-6 cycloalkyl" as used herein as a group or part of a group defines a saturated cyclic hydrocarbon radical having from 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

It will be apparent to those skilled in the art that S(=O) ₂ or _SO2 represent a sulfonyl moiety.

It will be clear to one skilled in the art that CO or C(=O) represents a carbonyl moiety.

Groups like -CRR-

を表すことは、当業者にとって明らかであろう。そのような基の例は、－ＣＲ^５ａＲ^５ｂ－である。

It will be clear to a person skilled in the art that this group represents an example of such a group is -CR ^5a R ^5b -.

Groups like -NR-

を表すことは、当業者にとって明らかであろう。そのような基の例は、－ＮＲ^５ｃ－である。

It will be clear to those skilled in the art that this group represents an example of such a group is -NR ^5c -.

Non-limiting examples of "monocyclic 5- or 6-membered aromatic rings containing 1, 2, or 3 nitrogen atoms and optionally a carbonyl moiety" include, but are not limited to, pyrazolyl, imidazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, or 1,2-dihydro-2-oxo-4-pyridinyl.

Those skilled in the art will recognize the term as a 5- or 6-membered monocyclic aromatic ring containing 1, 2, or 3 nitrogen atoms and a carbonyl moiety,

が挙げられるが、これらに限定されないことを理解するであろう。

It will be understood that these include, but are not limited to:

When any variable occurs more than once in any component, each definition is independent.

When any variable occurs more than one time in any formula (e.g., formula (I)), each definition is independent.

In general, whenever the term "substituted" is used in the present invention, unless otherwise indicated or apparent from the context, it is meant to indicate that one or more hydrogens, particularly 1 to 4 hydrogens, more particularly 1 to 3 hydrogens, preferably 1 or 2 hydrogens, more preferably 1 hydrogen, on the atom or radical shown in the expression are replaced with a selection from the indicated group, provided that the normal valence is not exceeded, and that the replacement results in a chemically stable compound, i.e., a compound that is sufficiently robust to withstand isolation from the reaction mixture to a useful purity (post-reaction isolation, e.g., purification by silica gel chromatography). In certain embodiments, when the number of substituents is not explicitly specified, the number of substituents is 1.

Combinations of substituents and/or variables are permissible only if such combinations result in chemically stable compounds. "Stable compound" in this context is meant to indicate a compound that is sufficiently robust to survive isolation from a reaction mixture to a useful degree of purity (post-reaction isolation, e.g., purification by silica gel chromatography).

Those skilled in the art will understand that the term "optionally substituted" means that the atom or radical designated in the expression using "optionally substituted" may be substituted or unsubstituted (which means substituted or unsubstituted, respectively).

When two or more substituents are present on a moiety, they may replace hydrogen atoms on the same atom or they may replace hydrogen atoms on different atoms in the moiety, unless otherwise indicated or clear from the context.

Within the context of the present invention, "saturated" means "fully saturated" unless otherwise specified.

Unless otherwise stated or apparent from the context, the aromatic ring group may be attached to the remainder of the molecule of formula (I) by any available ring carbon atom (C-bond) or nitrogen atom (N-bond).

Unless otherwise specified or apparent from the context, aromatic ring groups may be optionally substituted on carbon and/or nitrogen atoms, where possible, according to the embodiment.

As used herein, the term "comprising" encompasses the terms "consisting of" and "consisting essentially of." All embodiments described herein using the term "comprising" are also applicable to embodiments of the invention where the term "comprising" is limited to "consisting essentially of." Similarly, all embodiments described herein using the term "comprising" are also applicable to embodiments of the invention where the term "comprising" is limited to "consisting essentially of."

As used herein, the term "subject" refers to an animal, preferably a mammal (e.g., a cat, a dog, a primate, or a human), more preferably a human, who is or has been the object of treatment, observation, or experiment.

The term "therapeutically effective amount," as used herein, means an amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician, including the alleviation or reversal of symptoms of the disease or disorder being treated.

The term "composition" is intended to encompass a product containing specified ingredients in specified amounts, and any product resulting directly or indirectly from a combination of specified ingredients in specified amounts.

As used herein, the terms "treatment" and "treating" are intended to refer to any process that may slow, interrupt, halt or stop the progression of a disorder or improve one or more symptoms thereof, but does not necessarily indicate the complete elimination of all symptoms.

As used herein, any chemical formula with bonds shown only as solid lines and not as solid wedge bonds or hashed wedge bonds, or otherwise shown as having a particular configuration (e.g., R, S) around one or more atoms contemplates each possible stereoisomer, or a mixture of two or more stereoisomers.

In the above and below, the term "compound of formula (I)" is meant to include its tautomers and its stereoisomeric forms.

In the above and below, the term "compound of formula (Z)" is meant to include its tautomers and its stereoisomeric forms.

The terms "stereoisomer", "stereoisomeric form" or "stereochemically isomeric form" above and below are used interchangeably.

The present invention includes all stereoisomers of the compounds of the present invention, either as pure stereoisomers or as mixtures of two or more stereoisomers.

Enantiomers are stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a racemate or racemic mixture.

Atropisomers (or atropoisomers) are stereoisomers with specific spatial configurations resulting from restricted rotation about a single bond due to significant steric hindrance. All atropisomers of the compounds of formula (I) are intended to be included within the scope of the present invention.

Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, i.e., they are not related as mirror images. If the compound contains a double bond, the substituents may be in the E or Z configuration.

Substituents on a divalent cyclic saturated or partially saturated radical can have either a cis or trans configuration; for example, if the compound contains a disubstituted cycloalkyl group, the substituents can be in the cis or trans configuration.

The present invention therefore includes, whenever chemically possible, enantiomers, atropisomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers, and mixtures thereof.

The meanings of all terms, i.e. enantiomers, atropisomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof, are known to the skilled artisan.

Absolute configuration is specified according to the Cahn-Ingold-Prelog system. The configuration at the asymmetric atom is specified by either R or S. Resolved stereoisomers whose absolute configuration is not known can be designated (+) or (-) depending on the direction they rotate plane polarized light. For example, resolved enantiomers whose absolute configuration is not known can be designated (+) or (-) depending on the direction they rotate plane polarized light.

When a particular stereoisomer is specified, this means that the stereoisomer is substantially free of other stereoisomers, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, particularly less than 2%, most preferably less than 1% of other stereoisomers. Thus, when a compound of formula (I) is specified, for example, as (R), this means that the compound is substantially free of the (S) isomer, when a compound of formula (I) is specified, for example, as E, this means that the compound is substantially free of the Z isomer, and when a compound of formula (I) is specified, for example, as cis, this means that the compound is substantially free of the trans isomer.

Some of the compounds according to formula (I) may also exist in their tautomeric forms. Such forms, to the extent that they may exist, are intended to be included within the scope of the present invention even if not explicitly shown in formula (I) above. It follows that a single compound may exist in both stereoisomeric and tautomeric forms.

Pharmaceutically acceptable salts include acid addition salts and base addition salts. Such salts can be formed by conventional means, for example, by reacting the free acid or free base form with one or more equivalents of a suitable base or acid, optionally in a solvent or medium in which the salt is insoluble, followed by removal of the solvent or medium using standard techniques (e.g., in vacuum, by lyophilization, or by filtration). Salts can also be prepared by exchanging a counterion of a compound of the present disclosure in the form of a salt for another counterion, for example, using a suitable ion exchange resin.

The pharma- ceutically acceptable salts referred to above and below are meant to include the salt forms of therapeutically active non-toxic acids and bases which the compounds of formula (I) and their solvates are able to form.

Suitable acids include inorganic acids such as, for example, hydrohalic acids, e.g., hydrochloric or hydrobromic acids, sulfuric, nitric, phosphoric acids, and the like, or organic acids such as, for example, acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, oxalic acid (i.e., ethanedioic acid), malonic acid, succinic acid (i.e., butanedioic acid), maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid, pamoic acid, and the like. Conversely, the above salt forms can be converted to the free salt form by treatment with an appropriate base.

Compounds of formula (I) or solvates thereof containing acidic protons may be converted to their non-toxic metal or amine salt forms by treatment with appropriate organic and inorganic bases.

Suitable base salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts such as lithium, sodium, potassium, cesium, magnesium, calcium salts, and the like, organic bases such as primary, secondary and tertiary aliphatic and aromatic amines such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline, and isoquinoline, benzathine, N-methyl-glucamine, hydrabamine salts, and salts with amino acids such as arginine, lysine, and the like. Conversely, the base forms can be converted to the free base forms by treatment with acid.

The term "prodrug" includes any compound that, following oral or parenteral administration, in particular following oral administration, is metabolized to a (more) active form in an experimentally detectable amount and within a given time (e.g. within a 0.5-24 hour dosing interval, or for example within a 6-24 hour dosing interval (i.e. once to four times daily)). For the avoidance of doubt, the term "parenteral" administration includes all forms of administration other than oral administration, in particular intravenous (IV), intramuscular (IM) and subcutaneous (SC) injection.

Prodrugs can be prepared by modifying functional groups present on a compound such that the modification is cleaved in vivo when such prodrug is administered to a mammalian subject. The modification is typically accomplished by synthesizing the parent compound with a prodrug substituent. In general, prodrugs include compounds in which a hydroxyl, amino, sulfhydryl, carboxy, or carbonyl group is bonded to any group that can be cleaved in vivo to regenerate the free hydroxyl, amino, sulfhydryl, carboxy, or carbonyl group, respectively.

Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxyl functional groups, ester groups of carboxyl functional groups, N-acyl derivatives, and N-Mannich bases. General information on prodrugs can be found, for example, in Bundegaard, H. "Design of Prodrugs" p. 1-92, Elesevier, New York-Oxford (1985).

The term solvates includes the solvent addition forms that the compounds of formula (I) can form as well as the salts thereof. Examples of such solvent addition forms are, for example, hydrates, alcoholates, etc.

The compounds of the invention prepared by the processes described below may be synthesized in the form of mixtures of enantiomers, in particular racemic mixtures of enantiomers, which may be separated from one another according to resolution procedures known in the art. Methods for separating the enantiomeric forms of the compounds of formula (I) and their pharma- ceutical acceptable salts and solvates include liquid chromatography using chiral stationary phases. Said pure stereochemically isomers may also be derived from the corresponding pure stereochemically isomers of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably, when a particular stereoisomer is desired, said compound will be synthesized by stereospecific preparative methods. These methods will advantageously employ enantiomerically pure starting materials.

As used herein, the term "enantiomerically pure" means that the product contains at least 80% by weight of one enantiomer and no more than 20% by weight of the other enantiomer. Preferably, the product contains at least 90% by weight of one enantiomer and no more than 10% by weight of the other enantiomer. In the most preferred embodiment, the term "enantiomerically pure" means that the composition contains at least 99% by weight of one enantiomer and no more than 1% of the other enantiomer.

The present invention also includes isotopically labeled compounds that are identical to those listed herein, but due to the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature (or the most abundant atom found in nature).

All isotopes and isotopic mixtures of any particular atom or element identified herein, whether naturally occurring or synthetically produced, either at natural abundance or in isotopically enriched form, are contemplated within the scope of the present invention. Exemplary isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as ^2H , ^3H , ^11C , ^13C , ^14C , ^13N , ^15O , ^{17O ,} 18O, ^32P , ^33P , ^35S , ^18F , ^36Cl , ^122I , ^123I , ^125I , ^131I , ^75Br , ^76Br , ^77Br , and ^82Br . Preferably, the isotope is selected from the group of ^2H , ^3H , ^11C , ^13C , and ^18F . Preferably, the isotope is selected from the group of ^2H , ^3H , ^11C , and ^18F . More preferably, the isotope is ^2H , ^3H , or ^13C . More preferably, the isotope is ^2H or ^13C . More preferably, the isotope is ^2H . In particular, deuterated and ^13C -enriched compounds are intended to be included within the scope of the present invention. In particular, deuterated compounds are intended to be included within the scope of the present invention.

Certain isotopically labeled compounds (e.g., those labeled with ³ H and ¹⁴ C) may be useful, for example, in substrate tissue distribution assays. Tritiated ( ³ H) and carbon-14 ( ¹⁴ C) isotopes are useful for their ease of preparation and detectability. Additionally, substitution with heavier isotopes, such as deuterium (i.e., ² H), may afford greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and therefore may afford certain therapeutic advantages and therefore may be preferred in some circumstances. For example, positron emitting isotopes such as ¹⁵ O, ¹³ N, ¹¹ C, and ¹⁸ F are useful for positron emission tomography (PET) studies. PET imaging in cancer finds utility in helping to localize and identify tumors, stage disease, and determine appropriate treatments. Human cancer cells overexpress a number of receptors or proteins that are potential disease-specific molecular targets. Radiolabeled tracers that bind with high affinity and specificity to such receptors or proteins on tumor cells have great potential for diagnostic imaging and targeted radionuclide therapy (Charron, Carlie L. et al. Tetrahedron Lett. 2016, 57(37), 4119-4127). Furthermore, target-specific PET radiotracers can be used as biomarkers to investigate and evaluate pathology, for example, by measuring target expression and treatment response (Austin R. et al. Cancer Letters (2016), doi:10.1016/j.canlet.2016.05.008).

Solid oral dosage forms, e.g., tablets or capsules, containing one or more compounds described herein may be administered in at least one dosage form at a time, if desired. The compounds may also be administered in sustained release formulations.

Additional oral forms in which the compounds described herein may be administered include elixirs, solutions, syrups, and suspensions, each optionally containing flavoring and coloring agents.

Alternatively, one or more compounds described herein may be administered by inhalation (intramedial or intranasal) or in the form of a suppository or pessary, or may be applied topically in the form of a lotion, solution, cream, ointment, or dusting powder. For example, the compounds can be added to a cream comprising, consisting of, and/or consisting essentially of an aqueous emulsion of polyethylene glycol or liquid paraffin. The compounds can also be added to an ointment comprising, consisting of, and/or consisting essentially of a wax or soft paraffin base, together with optional stabilizers and preservatives as required, at a concentration of about 1% to about 10% by weight of the cream. Alternative means of administration include transdermal administration by use of a skin or transdermal patch.

The pharmaceutical compositions used in the methods of the present invention (as well as the compounds alone) can be injected parenterally, for example, intracavernosally, intravenously, intramuscularly, subcutaneously, intradermally, or intrathecally. In this case, the compositions also include at least one of a suitable carrier, a suitable excipient, and a suitable diluent.

For parenteral administration, the pharmaceutical compositions of the invention are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts and simple sugars to make the solution isotonic with blood.

For buccal or sublingual administration, the pharmaceutical compositions of the present invention may be administered in the form of tablets or lozenges, which may be formulated in a conventional manner.

As a further example, a pharmaceutical composition containing a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and, optionally, at least one other anti-neoplastic agent as an active ingredient, can be prepared by mixing the compound with a pharma- ceutically acceptable carrier, a pharma- ceutically acceptable diluent, and/or a pharma- ceutically acceptable excipient according to conventional pharmaceutical compounding techniques. The carriers, excipients, and diluents can take a wide variety of forms, depending on the desired route of administration (e.g., oral, parenteral, etc.). Thus, for liquid oral formulations such as suspensions, syrups, elixirs, and solutions, suitable carriers, excipients, and diluents include water, glycols, oils, alcohols, flavoring agents, preservatives, stabilizers, colorants, and the like. For solid oral formulations such as powders, capsules, and tablets, suitable carriers, excipients, and diluents include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrants, and the like. To control the primary site of absorption and disintegration, solid oral formulations can be optionally coated with substances such as sugars or enteric coated. For parenteral administration, carriers, excipients and diluents usually comprise sterile water, and other ingredients may be added to enhance the solubility and preservability of the composition. Injectable suspensions or solutions can also be prepared using aqueous carriers with appropriate additives, such as solubilizers and preservatives.

According to certain embodiments, the method of using a therapeutically effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and optionally at least one other anti-neoplastic agent, may include a dose range of about 0.1 mg to about 3000 mg, or any specific amount or range therein, particularly a dose range of about 1 mg to about 1000 mg, or any specific amount or range therein, of active ingredients, in a regimen of about 1 to about (4x) per day for an average (70 kg) human, although it will be apparent to one skilled in the art that the therapeutically effective amounts of the compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and optionally at least one other anti-neoplastic agent will vary depending on the disease, syndrome, condition, and disorder being treated.

One embodiment of the present invention relates to a method of using a pharmaceutical composition for oral administration comprising a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof in an amount of about 1 mg to about 500 mg. Advantageously, the compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof may be administered in a single daily dose, or the total daily dose may be administered in 2, 3 and (4x) divided doses per day.

The optimal dosage of the compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof to be administered can be readily determined and will vary according to the particular compound used, the mode of administration, the strength of the preparation, and the progression of the hematopoietic disorder. In addition, factors related to the particular subject being treated, such as the subject's sex, age, weight, diet, and time of administration, will necessitate adjustment of the dosage to achieve an appropriate therapeutic level and the desired therapeutic effect. The dosages given above are therefore exemplary of the average case. Of course, there may be individual cases in which higher or lower dosage ranges are effective, and these are within the scope of the present invention.

The compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof may be administered in any of the compositions and dosing regimens described above, or by compositions and dosing regimens established in the art, whenever the compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to a subject in need thereof for use.

One embodiment of the present invention relates to a method of using a pharmaceutical composition for intravenous or subcutaneous administration comprising a BCL-2 inhibitor in an amount of about 1 mg to about 500 mg. Advantageously, the BCL-2 inhibitor can be administered in a single daily dose, or the total daily dose can be administered in 2, 3 and (4x) divided doses per day.

The optimal dose of BCL-2 inhibitor to be administered can be readily determined and will vary with the particular compound used, the mode of administration, the strength of the formulation, and the progression of the disease, syndrome, condition, or disorder. In addition, factors related to the particular subject being treated, such as the subject's sex, age, weight, diet, and time of administration, will necessitate adjustment of the dose to achieve appropriate therapeutic levels and the desired therapeutic effect. The dosages given above are therefore exemplary of the average case. It will be understood that there may be individual instances in which higher or lower dosage ranges are effective, and these are within the scope of the present invention.

The BCL-2 inhibitor can be administered in any of the compositions and administration regimens described above, or using compositions and administration regimens established in the art, whenever a BCL-2 inhibitor is administered to a subject in need thereof.

As used herein, the term "menin-MLL inhibitors" refers to inhibitors of the protein-protein interaction between menin and mixed lineage leukemia 1 (MLL1), also known in the scientific art as histone-lysine N-methyltransferase 2A (KMT2A) protein (UniProt Accession No. Q03164), which inhibit or reduce menin-MLL1 activity. The menin-MLL inhibitors described herein are disclosed in International Application No. CN2020/137266 (published as WO2021/121327 on June 24, 2021), which is incorporated herein by reference in its entirety, and which also disclose the corresponding synthesis schemes and analytical characterization.

As used herein, the term "BCL-2 inhibitor" refers to an agent that inhibits or reduces BCL-2 activity.

As used herein, the term "antineoplastic agent" refers to any agent that treats cancer.

As used herein, the term "hypomethylating agent" refers to an agent that inhibits or reduces DNA methylation.

As used herein, the term "kinase inhibitor" refers to an agent that inhibits or reduces the activity of at least one kinase (e.g., tyrosine and/or serine kinases, such as fms-like receptor tyrosine kinase-3 (FLT3), Bruton's tyrosine kinase (BTK), Abelson tyrosine kinase 1 (ABL), Auroracerine/tyrosine kinase, etc.).

As used herein, the term "FLT-3 inhibitors" refers to tyrosine kinase inhibitors (TKIs) that are classified into first-generation and next-generation inhibitors based on their potency and specificity for fms-like receptor tyrosine kinase-3 (FLT3) and their associated downstream targets.

As used herein, the term "CD20 inhibitor" refers to any agent that reduces the activity of CD20.

As used herein, the term "isocitrate dehydrogenase (IDH) inhibitor" refers to any agent that prevents the conversion of isocitrate to α-ketoglutarate (α-KG) in the tricarboxylic acid (TCA) cycle.

As used herein, the term "immunomodulatory anti-neoplastic agent" refers to any agent that enhances anti-tumor immune cell activity.

As used herein, the term "programmed cell death protein 1 (PD-1) inhibitor" refers to any agent that inhibits or reduces PD-1 activity.

As used herein, the term "dihydroorotate dehydrogenase (DHODH) inhibitor" refers to any agent that inhibits or reduces dihydroorotate dehydrogenase activity.

As used herein, unless otherwise indicated, the terms "affected" or "affected" (when referring to a disease, disorder, or medical condition affected by inhibition or alteration of menin-MLL activity) include a reduction in the frequency and/or severity of one or more symptoms or signs of said hematopoietic disorder and/or include prevention of the onset of one or more symptoms or signs of said hematopoietic disorder or prevention of the onset of a hematopoietic disorder.

As used herein, the term "hematopoietic disorder" refers to any disorder associated with the production of the cellular components of blood and plasma, including, but not limited to, blood cancers.

According to one embodiment, the present invention provides a combination as described herein.

According to one embodiment, the present invention provides a combination as described herein for use as a medicament.

According to one embodiment, the present invention provides a combination as described herein for the manufacture of a medicament.

According to one embodiment, the present invention provides a combination as described herein for the manufacture of a medicament for the treatment or prevention of any one of the disease conditions referred to herein.

According to one embodiment, the present invention provides a combination as described herein for use in the prevention or treatment, particularly the treatment, of a disease as described herein.

According to one embodiment, the present invention provides a combination as described herein for use in the prevention or treatment, particularly the treatment, of a hematopoietic disorder, including but not limited to, a blood cancer, including but not limited to, lymphoma, myeloma and leukemia.

According to one embodiment, the present invention provides a combination as described herein for use in the prevention or treatment, in particular the treatment, of a hematopoietic disorder.

According to one embodiment, the hematopoietic disorder is selected from, but is not limited to, lymphoma, myeloma, myelodysplasia, and leukemia.

According to one embodiment, the hematopoietic disorder is a lymphoma selected from Hodgkin's lymphoma and non-Hodgkin's lymphoma.

According to one embodiment, the lymphoma is a non-Hodgkin's lymphoma that is Burkitt's lymphoma, anaplastic large cell lymphoma, splenic marginal zone lymphoma, hepatosplenic T-cell lymphoma, or angioimmunoblastic T-cell lymphoma (AILT).

In one embodiment, the hematopoietic disorder is myeloma. In one embodiment, the hematopoietic disorder is multiple myeloma, Waldenstrom's macroglobulinemia, or plasmacytoma.

According to one embodiment, the hematopoietic disorder is myelodysplasia, including but not limited to myelodysplastic syndrome (MDS).

According to one embodiment, the hematopoietic disorder is leukemia.

According to one embodiment, the hematopoietic disorder is a leukemia selected from acute leukemia and chronic leukemia. According to one embodiment, the leukemia is acute leukemia. According to one embodiment, the leukemia is chronic leukemia.

According to one embodiment, the hematopoietic disorder is myeloid leukemia, myelogenous leukemia, lymphoblastic leukemia, or lymphocytic leukemia. According to one embodiment, the hematopoietic disorder is a leukemia selected from, but not limited to, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), small lymphocytic leukemia (SLL), acute myeloid leukemia (AML), chronic idiopathic myelofibrosis (MF), chronic myelogenous leukemia (CML), T-cell prolymphocytic leukemia (T-PLL), B-cell prolymphocytic leukemia (B-PLL), chronic neutrophilic leukemia (CNL), hairy cell leukemia (HCL), T-cell large granular lymphocytic leukemia (T-LGL), and aggressive NK cell leukemia. According to one embodiment, the AML is acute megakaryoblastic leukemia (AMKL).

According to one embodiment, the leukemia is MDS, CLL, SLL, ALL or AML. According to one embodiment, the leukemia is CLL, SLL or AML. According to one embodiment, the leukemia is CLL or SLL. In some embodiments, the CLL or SLL is a CD20 expressing cancer. According to one embodiment, the leukemia is ALL or AML. According to one embodiment, the leukemia is ALL. According to one embodiment, the leukemia is AML. According to one embodiment, the hematopoietic disorder is Waldenstrom's macroglobulinemia.

According to one embodiment, the hematopoietic disorder is MLL-rearranged leukemia, MLL partial tandem duplication (PTD) leukemia, MLL-amplified leukemia, MLL-positive leukemia, or leukemia exhibiting an elevated HOX/MEIS1 gene expression signature.

In one embodiment, the leukemia is MLL-rearranged leukemia and/or nucleophosmin 1 (NPM1) mutant leukemia. In one embodiment, the hematopoietic disorder is MLL-rearranged leukemia.

According to one embodiment, the hematopoietic disorder is nucleophosmin 1 (NPM1) mutant leukemia (e.g., NPM1c).

According to one embodiment, the present invention provides a method for treating a hematopoietic disorder, which is myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL), comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and optionally at least one other anti-neoplastic agent.

According to one embodiment, the hematopoietic disorder is myelodysplastic syndrome (MDS) or myeloproliferative neoplasm (MPN).

According to one embodiment, the hematopoietic disorder is acute lymphoblastic leukemia (ALL).

According to one embodiment, the hematopoietic disorder is acute myeloid leukemia (AML).

According to one embodiment, the hematopoietic disorder is small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL).

In one embodiment, the hematopoietic disorder is SLL or CLL, where SLL or CLL is a CD20-expressing cancer.

According to one embodiment, the hematopoietic disorder is myelodysplastic syndrome (MDS).

According to one embodiment, the hematopoietic disorder is a myeloproliferative neoplasm (MPN).

According to one embodiment, the hematopoietic disorder is NPM1 mutant leukemia with a FLT3 mutation.

According to one embodiment, the hematopoietic disorder is FLT3-dependent leukemia.

According to one embodiment, the hematopoietic disorder is MEF2G-dependent leukemia.

In one embodiment, the hematopoietic disorder has one or more MLL1 (KMT2A) gene rearrangements or alterations (e.g., duplications or amplifications) and/or NPM1 mutations.

In one embodiment, the hematopoietic disorder has (i) one or more MLL1 (KMT2A) gene rearrangements or alterations (e.g., duplications or amplifications) and/or NPM1 mutations, as well as (ii) a FLT3 mutation.

According to one embodiment, the hematopoietic disorder is MLL-rearranged leukemia.

According to one embodiment, the hematopoietic disorder is acute myeloid leukemia (AML).

According to one embodiment, the hematopoietic disorder is small lymphocytic lymphoma (SLL).

According to one embodiment, the hematopoietic disorder is chronic lymphocytic leukemia (CLL).

According to one embodiment, the hematopoietic disorder is acute leukemia, chronic leukemia, myeloid leukemia, myelogenous leukemia, lymphoblastic leukemia, lymphocytic leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), T-cell prolymphocytic leukemia (T-PLL), large granular lymphocytic leukemia, hairy cell leukemia (HCL), MLL-rearranged leukemia, MLL-PTD leukemia, MLL-amplified leukemia, MLL-positive leukemia, or leukemia exhibiting an elevated HOX/MEIS1 gene expression signature.

According to one embodiment, the hematopoietic disorder is AML, specifically nucleophosmin (NPM1) mutated AML (i.e., NPM1 ^mut AML), more specifically abstract NPM1 mutated AML.

According to one embodiment, the hematopoietic disorder is an MLL-rearranged leukemia, in particular MLL-rearranged AML or ALL.

According to one embodiment, the hematopoietic disorder comprises an MLL gene alteration, and in particular the hematopoietic disorder is AML or ALL with an MLL gene alteration. In a particular embodiment, the MLL gene alteration is a duplication. In a particular embodiment, the MLL gene alteration is an amplification.

According to one embodiment, the hematopoietic disorder includes an NPM1 gene mutation and/or an MLL1 (also known as KMT2A) gene mutation.

According to one embodiment, the MLL1 gene mutation includes, but is not limited to, MLL1 gene rearrangement, duplication, or amplification.

According to one embodiment, the hematopoietic disorder is mixed lineage leukemia (MLL), MLL-associated leukemia, MLL-associated leukemia, MLL-positive leukemia, MLL-induced leukemia, MLL-associated leukemia, acute leukemia, chronic leukemia, myelodysplastic syndrome (MDS), or myeloproliferative neoplasms (MPNs).

All of the embodiments described herein relating to methods for treating hematopoietic disorders are also applicable for use in treating such hematopoietic disorders.

All of the embodiments described herein for use in treating a hematopoietic disorder are also applicable to methods for treating that hematopoietic disorder.

All of the embodiments described herein relating to methods for treating hematopoietic disorders are also applicable for use in the methods for treating hematopoietic disorders.

All embodiments described herein for use in a method for treating a hematopoietic disorder are also applicable to the method for treating the hematopoietic disorder.

In one embodiment, the present invention provides
a therapeutically effective amount of a menin-MLL inhibitor of formula (I) or a tautomer or stereoisomeric form thereof or a pharma- ceutical acceptable salt or solvate thereof;
a therapeutically effective amount of a BCL-2 inhibitor;
- Optionally, with a therapeutically effective amount of at least one other anti-neoplastic agent.

According to one embodiment, the compound of formula (I) has the structure:

を有するメニン－ＭＬＬ阻害剤、並びにその互変異体及び立体異性形態［式中、
Ｒ^１ａは、－Ｃ（＝Ｏ）－ＮＲ^ｘａＲ^ｘｂ、Ｈｅｔ、又は

and tautomeric and stereoisomeric forms thereof,
R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

Ｈｅｔは、１、２又は３個の窒素原子及び任意選択的にカルボニル部分を含有する５又は６員単環式芳香環を表し、
当該５又は６員単環式芳香環は、任意選択的に、Ｃ_３～６シクロアルキル及びＣ_１～４アルキルからなる群から選択される１又は２個の置換基で置換され、
Ｒ^ｘａ及びＲ^ｘｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^１ｂは、Ｆ又はＣｌを表し、
Ｙ^１は、－ＣＲ^５ａＲ^５ｂ－、－Ｏ－、又は－ＮＲ^５ｃ－を表し、
Ｒ^２は、水素、ハロ、Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－ＮＲ^７ａＲ^７ｂからなる群から選択され、
Ｕは、Ｎ又はＣＨを表し、
ｎ１、ｎ２、ｎ３及びｎ４は、１及び２からそれぞれ独立して選択され、
Ｘ^１はＣＨを表し、Ｘ^２はＮを表し、
Ｒ^４は、イソプロピルを表し、
Ｒ^５ａ、Ｒ^５ｂ、Ｒ^５ｃ、Ｒ^７ａ及びＲ^７ｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^３は、－Ｃ_１～６アルキル－ＮＲ^８ａＲ^８ｂ、－Ｃ_１～６アルキル－Ｃ（＝Ｏ）－ＮＲ^９ａＲ^９ｂ、－Ｃ_１～６アルキル－ＯＨ、又は－Ｃ_１～６アルキル－ＮＲ^１１－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル－Ｏ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルを表し、ここで、Ｒ^３の定義におけるＣ_１～４アルキル又はＣ_１～６アルキル部分のそれぞれは、互いに独立して、シアノ、ハロ、－ＯＨ、及び－Ｏ－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されていてもよく、
Ｒ^８ａ及びＲ^８ｂは、水素、Ｃ_１～６アルキル、－Ｃ（＝Ｏ）－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１２ａＲ^１２ｂ、並びに、－ＯＨ、シアノ、ハロ、－Ｓ（＝Ｏ）_２－Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１０ａＲ^１０ｂ及び－ＮＲ^１０ｃ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されているＣ_１～６アルキル、からなる群からそれぞれ独立して選択され、
Ｒ^９ａ、Ｒ^９ｂ、Ｒ^１０ａ、Ｒ^１０ｂ、Ｒ^１０ｃ、Ｒ^１１、Ｒ^１２ａ、及びＲ^１２ｂは、水素及びＣ_１～６アルキルからなる群からそれぞれ独立して選択される］、
並びにそれらの医薬的に許容される塩及び溶媒和物である。

Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety;
the 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl, wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ may be substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C _1-6 alkyl substituted with _1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) _{2 -C 1-4} alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O _{)-C 1-4} alkyl;
R ^9a , R ^9b , R ^10a , R ^10b , R ^10c , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

を表し、
Ｈｅｔは、１、２又は３個の窒素原子及び任意選択的にカルボニル部分を含有する５又は６員単環式芳香環を表し、
当該５又は６員単環式芳香環は、任意選択的に、Ｃ_３～６シクロアルキル及びＣ_１～４アルキルからなる群から選択される１又は２個の置換基で置換され、
Ｒ^ｘａ及びＲ^ｘｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^１ｂは、Ｆ又はＣｌを表し、
Ｙ^１は、－ＣＲ^５ａＲ^５ｂ－、－Ｏ－、又は－ＮＲ^５ｃ－を表し、
Ｒ^２は、水素、ハロ、Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－ＮＲ^７ａＲ^７ｂからなる群から選択され、
Ｕは、Ｎ又はＣＨを表し、
ｎ１、ｎ２、ｎ３及びｎ４は、１及び２からそれぞれ独立して選択され、
Ｘ^１はＣＨを表し、Ｘ^２はＮを表し、
Ｒ^４は、イソプロピルを表し、
Ｒ^５ａ、Ｒ^５ｂ、Ｒ^５ｃ、Ｒ^７ａ及びＲ^７ｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^３は、－Ｃ_１～６アルキル－ＮＲ^８ａＲ^８ｂ、－Ｃ_１～６アルキル－Ｃ（＝Ｏ）－ＮＲ^９ａＲ^９ｂ、－Ｃ_１～６アルキル－ＯＨ、又は－Ｃ_１～６アルキル－ＮＲ^１１－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル－Ｏ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルを表し、
ここで、Ｒ^３の定義におけるＣ_１～４アルキル又はＣ_１～６アルキル部分のそれぞれは、互いに独立して、シアノ、ハロ、又は－Ｏ－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されていてもよく、
Ｒ^８ａ及びＲ^８ｂは、水素、Ｃ_１～６アルキル、－Ｃ（＝Ｏ）－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１２ａＲ^１２ｂ、並びに、シアノ、ハロ、－Ｓ（＝Ｏ）_２－Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－Ｃ（＝Ｏ）－ＮＲ^１０ａＲ^１０ｂからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されているＣ_１～６アルキル、からなる群からそれぞれ独立して選択され、
Ｒ^９ａ、Ｒ^９ｂ、Ｒ^１０ａ、Ｒ^１０ｂ、Ｒ^１１、Ｒ^１２ａ、及びＲ^１２ｂは、水素及びＣ_１～６アルキルからなる群からそれぞれ独立して選択される］、
並びにそれらの医薬的に許容される塩及び溶媒和物である。

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety;
the 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl;
wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ is optionally substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, or -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C 1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O)-NR ^10a R ^10b _;
R ^9a , R ^9b , R ^10a , R ^10b , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

を表し、
Ｈｅｔは、１、２又は３個の窒素原子及び任意選択的にカルボニル部分を含有する５又は６員単環式芳香環を表し、
当該５又は６員単環式芳香環は、任意選択的に、Ｃ_３～６シクロアルキル及びＣ_１～４アルキルからなる群から選択される１又は２個の置換基で置換され、
Ｒ^ｘａ及びＲ^ｘｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^１ｂは、Ｆ又はＣｌを表し、
Ｙ^１は、－ＣＲ^５ａＲ^５ｂ－、－Ｏ－、又は－ＮＲ^５ｃ－を表し、
Ｒ^２は、水素、ハロ、Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－ＮＲ^７ａＲ^７ｂからなる群から選択され、
Ｕは、Ｎ又はＣＨを表し、
ｎ１、ｎ２、ｎ３及びｎ４は、１及び２からそれぞれ独立して選択され、
Ｘ^１はＣＨを表し、Ｘ^２はＮを表し、
Ｒ^４は、イソプロピルを表し、
Ｒ^５ａ、Ｒ^５ｂ、Ｒ^５ｃ、Ｒ^７ａ及びＲ^７ｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^３はＣ_１～６アルキル－ＮＲ^８ａＲ^８ｂを表し、ここで、Ｒ^３の定義におけるＣ_１～６アルキル部分は、シアノ、ハロ、－ＯＨ、及び－Ｏ－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されていてもよく、
Ｒ^８ａ及びＲ^８ｂは、水素、Ｃ_１～６アルキル、並びに、
－ＯＨ、シアノ、ハロ、－Ｓ（＝Ｏ）_２－Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、－Ｃ（＝Ｏ）－ＮＲ^１０ａＲ^１０ｂ、及び－ＮＲ^１０ｃ－Ｃ（＝Ｏ）－Ｃ_１～４アルキルからなる群から選択される１、２又は３個の置換基でそれぞれ独立して置換されているＣ_１～６アルキル、からなる群からそれぞれ独立して選択され、
Ｒ^１０ａ、Ｒ^１０ｂ、Ｒ^１０ｃは、水素及びＣ_１～６アルキルからなる群からそれぞれ独立して選択される］、
並びにそれらの医薬的に許容される塩及び溶媒和物である。

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety;
the 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl portion in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -O-C _1-4 alkyl;
R ^8a and R ^8b are hydrogen, C _1-6 alkyl, and
C _1-6 alkyl, each independently substituted with 1, 2 or 3 substituents selected from the group consisting of: -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

を表し、
Ｈｅｔは、１、２又は３個の窒素原子及び任意選択的にカルボニル部分を含有する５又は６員単環式芳香環を表し、
当該５又は６員単環式芳香環は、任意選択的に、Ｃ_３～６シクロアルキル及びＣ_１～４アルキルからなる群から選択される１又は２個の置換基で置換され、
Ｒ^ｘａ及びＲ^ｘｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^１ｂは、Ｆ又はＣｌを表し、
Ｙ^１は、－ＣＲ^５ａＲ^５ｂ－、－Ｏ－、又は－ＮＲ^５ｃ－を表し、
Ｒ^２は、水素、ハロ、Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－ＮＲ^７ａＲ^７ｂからなる群から選択され、
Ｕは、Ｎ又はＣＨを表し、
ｎ１、ｎ２、ｎ３及びｎ４は、１及び２からそれぞれ独立して選択され、
Ｘ^１はＣＨを表し、Ｘ^２はＮを表し、
Ｒ^４は、イソプロピルを表し、
Ｒ^５ａ、Ｒ^５ｂ、Ｒ^５ｃ、Ｒ^７ａ及びＲ^７ｂは、水素、Ｃ_１～４アルキル及びＣ_３～６シクロアルキルからなる群からそれぞれ独立して選択され、
Ｒ^３は、Ｃ_１～６アルキル－ＮＲ^８ａＲ^８ｂを表し、ここで、Ｒ^３の定義におけるＣ_１～６アルキル部分は、シアノ、ハロ、及び－Ｏ－Ｃ_１～４アルキルからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されていてもよく、
Ｒ^８ａ及びＲ^８ｂは、水素、Ｃ_１～６アルキル、並びに、シアノ、ハロ、－Ｓ（＝Ｏ）_２－Ｃ_１～４アルキル、－Ｏ－Ｃ_１～４アルキル、及び－Ｃ（＝Ｏ）－ＮＲ^１０ａＲ^１０ｂからなる群からそれぞれ独立して選択される１、２又は３個の置換基で置換されているＣ_１～６アルキル、からなる群からそれぞれ独立して選択され、
Ｒ^１０ａ及びＲ^１０ｂは、水素及びＣ_１～６アルキルからなる群からそれぞれ独立して選択される］、
並びにそれらの医薬的に許容される塩及び溶媒和物である。

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety;
the 5- or 6-membered monocyclic aromatic ring is optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl portion in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O)-NR ^10a R ^10b _;
R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb or Het;
Het represents a 6-membered monocyclic aromatic ring containing two nitrogen atoms, said 6-membered monocyclic aromatic ring being substituted with one C _3-6 cycloalkyl;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl;
wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ may be substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of -OH, and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C _1-6 alkyl substituted with _1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) _{2 -C 1-4} alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O _{)-C 1-4} alkyl;
R ^9a , R ^9b , R ^10a , R ^10b , R ^10c , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb or Het;
Het represents a 6-membered monocyclic aromatic ring containing two nitrogen atoms, said 6-membered monocyclic aromatic ring being substituted with one C _3-6 cycloalkyl;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl portion in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2 _{or 3} substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a is -C(=O)-NR ^xa R ^xb ,
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl portion in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2 _{or 3} substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb or Het;
Het represents pyrimidinyl substituted with one C _3-6 cycloalkyl;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl moiety in the definition of R ³ may be substituted by one -OH;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _1-6 alkyl substituted with 1 or 2 substituents each independently selected from the group consisting of halo, -O-C _1-4 alkyl, and -NR ^10c -C(═O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb or Het;
Het represents pyrimidinyl substituted with one C _3-6 cycloalkyl;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n2 is 2,
n1, n3 and n4 are 1;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents C _1-6 alkyl-NR ^8a R ^8b , where the C _1-6 alkyl moiety in the definition of R ³ may be substituted by one -OH;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _1-6 alkyl substituted with 1 or 2 substituents each independently selected from the group consisting of halo, -O-C _1-4 alkyl, and -NR ^10c -C(═O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a is -C(=O)-NR ^xa R ^xb ,
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n2 is 2,
n1, n3 and n4 are 1;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _C1-6 alkyl ^{- NR8aR8b} ;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _1-6 alkyl substituted with 1 or 2 substituents each independently selected from the group consisting of halo, -O-C _1-4 alkyl, and -NR ^10c -C(═O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n2 is 2,
n1, n3 and n4 are 1;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _CH2 - _CH2 - _CH2- ^{NR8aR8b ;}
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _1-6 alkyl substituted with 1 or 2 substituents each independently selected from the group consisting of halo, -O-C _1-4 alkyl, and -NR ^10c -C(═O)-C _1-4 alkyl;
R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _C1-6 alkyl ^{- NR8aR8b} ;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2, or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O) ^{-NR 10a} R ^10b _;
R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _CH2 - _CH2 - _CH2- ^{NR8aR8b ;}
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2, or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O) ^{-NR 10a} R ^10b _;
R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent hydrogen or C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _CH2 - _CH2 - _CH2- ^{NR8aR8b ;}
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2, or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O) ^{-NR 10a} R ^10b _;
R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent hydrogen or C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _CH2 - _CH2 - _CH2- ^{NR8aR8b ;}
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _1-6 alkyl substituted with 1, 2, or 3 substituents each independently selected from the group consisting of -OH and -O-C _1-4 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _C1-6 alkyl ^{- NR8aR8b} ;
R ^8a and R ^8b are each independently selected from the group consisting of C _1-6 alkyl, and C _1-6 alkyl substituted with one —O—C _1-4 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 represents hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
^R3 represents _CH2 - _CH2 - _CH2- ^{NR8aR8b ;}
R ^8a and R ^8b are each independently selected from the group consisting of C _1-6 alkyl, and C _1-6 alkyl substituted with one —O—C _1-4 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb or Het;
Het represents a 6-membered monocyclic aromatic ring containing two nitrogen atoms, said 6-membered monocyclic aromatic ring being optionally substituted with one C _3-6 cycloalkyl;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 is hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C 1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) _{2 -C 1-4 alkyl} , and -O-C _1-4 alkyl;
R ^9a , R ^9b , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

According to one embodiment, the compounds of formula (I) are as defined herein, as well as their tautomeric and stereoisomeric forms,
R ^1a represents -C(=O)-NR ^xa R ^xb ;
R ^xa and R ^xb represent C _1-4 alkyl;
R ^1b represents F;
Y ¹ represents -O-;
^R2 is hydrogen;
U represents N;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b or -C _1-6 alkyl-OH;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C 1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) _{2 -C 1-4 alkyl} , and -O-C _1-4 alkyl;
R ^9a , R ^9b , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl;
and pharma-ceutically acceptable salts and solvates thereof.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R1b represents F.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R2 represents hydrogen.

In one embodiment, the invention comprises a compound of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein n1 is 1, n2 is 2, n3 is 1, and n4 is 1.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^Y1 represents —O—.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^Y1 represents -O- and U represents N.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^Y1 represents -O-, U represents N, ^R1b represents F and ^R2 represents hydrogen.

In one embodiment, the present invention comprises a compound of formula (I), as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Het is

を表す。

Represents.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Het represents a monocyclic 5- or 6-membered aromatic ring containing 1 or 2 nitrogen atoms,
wherein said monocyclic 5- or 6-membered aromatic ring is substituted with one C _3-6 cycloalkyl.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Het represents a monocyclic 5- or 6-membered aromatic ring containing 1 or 2 nitrogen atoms, said monocyclic 5- or 6-membered aromatic ring being substituted with one _{C3-6cycloalkyl} , and ^R1b represents F.

In one embodiment, the invention comprises a compound of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Het represents a monocyclic 6-membered aromatic ring containing 1 or 2 nitrogen atoms, said monocyclic 6-membered aromatic ring being substituted with one _{C3-6cycloalkyl} .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein Het represents a monocyclic 6-membered aromatic ring containing 1 or 2 nitrogen atoms, said monocyclic 6-membered aromatic ring being substituted with one _{C3-6cycloalkyl} , and ^R1b represents F.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl - ^NR8aR8b , wherein the ^C1-6alkyl moiety in the definition of ^R3 is optionally substituted by 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo, and -O- _C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl - ^NR8aR8b , wherein the ^C1-6alkyl moiety in the definition of ^R3 is optionally substituted by 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O- _C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl ^{- NR8aR8b} .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents -Ci_6alkyl-NR ^8a R ^8b , wherein the _{Ci_6alkyl} moiety in the definition of R ³ is optionally substituted with 1 _{, 2} or 3 substituents each independently selected from the group consisting of cyano, halo, and -O- _{Ci_4alkyl} , and R ^8a and R ^8b are each independently selected from the group consisting of _hydrogen , _{Ci_6alkyl, and Ci_6alkyl} substituted with ₁ , 2 or ₃ substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) 2 -Ci_4alkyl _{, -O-} Ci_4alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O) _{-Ci_4alkyl} .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _{-Ci_6alkyl} - ^NR8aR8b , the ^C1-6alkyl moiety in the definition of R3 is optionally substituted with 1, 2 or ³ substituents each independently selected from the group consisting of cyano, ^halo , -OH and -O-Ci_4alkyl, and _R8a and ^R8b are each independently selected from the group consisting of hydrogen, _C1-6alkyl and _C1-6alkyl substituted with 1 _{, 2 or 3 substituents} each independently selected from the group consisting of -OH, ^cyano , halo, -S(=O) ₂ -Ci_4alkyl, -O-Ci_4alkyl, -C(=O) ^-NR10aR10b and ^-NR10c _- C(=O)-Ci_4alkyl.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , wherein the C _1-6 alkyl moiety in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, and -O-C _1-4 alkyl, and R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C 1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, and -C(=O)-NR ^10a R ^10b _.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents -Ci_6alkyl- ^NR8aR8b , wherein the ^C1-6alkyl moiety in the definition of ^R3 is optionally substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo, and -O- _{Ci_4alkyl} , and ^R8a and ^R8b are each independently selected from the group consisting of hydrogen, _C1-6alkyl , and _C1-6alkyl substituted with 1, _{2 or 3} substituents each independently selected from the group consisting of cyano, halo, -S(=O) _2- Ci_4alkyl, -O _- ^Ci_4alkyl , -C(=O) ^-NR10aR10b and ^-NR10c -C(=O)-Ci_4alkyl.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents -Ci- _6alkyl -NR ^8a R ^8b , wherein the Ci _-6alkyl moiety in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-Ci _- 4alkyl, and R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, Ci _- 6alkyl and Ci-6alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) _{2 -Ci- 4alkyl} , -O-Ci- _4alkyl and -C(=O)-NR ^10a R ^10b.

In one embodiment, the invention comprises compounds of formula (I) as defined in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents -Ci- _6alkyl -NR ^8a R ^8b , wherein the Ci _-6alkyl moiety in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-Ci _-4alkyl , and R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, Ci _{-6alkyl and Ci-6alkyl} substituted with 1 _{, 2} or ₃ substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -Ci-4alkyl, -O-Ci-4alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O)-Ci _{- 4alkyl} .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C2-6alkyl - ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted by 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo and -O- _C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents -C2-6alkyl- ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted by 1, ₂ or ₃ substituents each independently selected from the group consisting of cyano, halo, -OH and -O- _C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C2-6alkyl - ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo and -O- _C1-4alkyl , and ^R8a and ^R8b are each independently selected from the group consisting of hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with ₁ , 2 or ₃ substituents each independently selected from the group consisting of -OH, ^cyano , halo, -S(=O) ₂ -C1-4alkyl, -O- _C1-4alkyl , -C(=O) ^-NR10aR10b and ^-NR10c -C(=O) _-C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents _-C2-6alkyl -NR ^8a R ^8b , wherein the C2-6alkyl moiety in the definition of R ³ is optionally substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O- _C1-4alkyl , and R ^8a and R ^8b represent hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ - _C1-4alkyl , -O- _C1-4alkyl , -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O) _{-C1-4alkyl. 1-6} alkyl.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents -C2-6alkyl- ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted with 1, ₂ or ₃ substituents each independently selected from the group consisting of cyano, halo, -OH and -O- _C1-4alkyl , and ^R8a and ^R8b are each independently selected from the group consisting of hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) _{2 -C1-4alkyl ,} -O- _C1-4alkyl and -C(=O) ^{-NR10aR10b .}

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C2-6alkyl - ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo and -O- _C1-4alkyl , and ^R8a and ^R8b are each independently selected from the group consisting of hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C1-4alkyl, -O _{- C1-4alkyl} and -C(=O) ^{-NR10aR10b .}

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl ^{- NR8aR8b} ;
R ^8a and R ^8b are each independently selected from the group consisting of C _{1-6 alkyl and C 1-6} alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _{1-4 alkyl} , and -C(=O)-NR ^10a R ^10b .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein R ³ represents _-C2-6alkyl -NR ^8a R ^8b , wherein the _C2-6alkyl moiety in the definition of R ³ is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-C1-4alkyl, and R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with ₁ , ₂ or ₃ substituents each independently selected from the group consisting of cyano, halo, -S(=O) _2- C1-4alkyl, -O-C1-4alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c _- C(=O) _-C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C2-6alkyl - ^NR8aR8b , wherein the ^C2-6alkyl moiety in the definition of ^R3 is optionally substituted with 1, ₂ or 3 substituents each independently selected from the group consisting of cyano, halo and -O- _C1-4alkyl , and ^R8a and ^R8b are each independently selected from the group consisting of hydrogen, _{C1-6alkyl and C1-6alkyl} substituted with 1, ₂ or ₃ substituents each independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C1-4alkyl, -O _- ^{C1-4alkyl, -C(=O)-NR10aR10b and -NR10c} -C(=O) _-C1-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl ^{- NR8aR8b} ;
R ^8a and R ^8b are each independently selected from the group consisting of C _{1-6 alkyl and C 1-6} alkyl substituted with 1, 2 _{or 3} substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)-C _1-4 alkyl.

In one embodiment, the invention comprises compounds of formula (I) as described in any of the other embodiments, and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-C1-6alkyl - ^NR8aR8b , ^R8a represents _C1-6alkyl and ^R8b represents ^C1-6alkyl substituted by _one -O- _C1-4alkyl .

In one embodiment, the present invention relates to a method of use of the compounds of formula (I) and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof as described in any of the other embodiments, wherein ^R3 represents -Ci _-6alkyl ^{- NR8aR8b} , -Ci _-6alkyl -C(=O) ^-NR9aR9b , -Ci _-6alkyl -OH or -Ci _-6alkyl - ^NR11 -C(=O)-O-Ci _- 4alkyl-O-C ⁽ =O)-Ci _- 4alkyl, wherein each of the Ci _- 4alkyl or Ci _-6alkyl moieties in the definition of ^R3 , independently of each other, is optionally substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo or -O-Ci _-4alkyl .

In one embodiment, the invention comprises compounds of formula (I) and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof as described in any of the other embodiments, wherein ^R3 represents -Ci _-6alkyl - ^NR8aR8b , -Ci-6alkyl-C(= ^{O)-NR9aR9b or -Ci} _-6alkyl - _NR11 ^- C(=O)-O-Ci-4alkyl-O-C(=O)-Ci _- 4alkyl, wherein each of the Ci _- 4alkyl or Ci _-6alkyl moieties in the ^R3 definition, independently of each other, is optionally substituted by _1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-Ci- _4alkyl .

In one embodiment, the invention relates to those compounds of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-CH2 - _CH2 - _CH2 ^{- NR8aR8b} .

In one embodiment, the invention relates to those compounds of formula (I) as described in any of the other embodiments, and the pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof, wherein ^R3 represents _-CH2 - _CH2 ^{-CH2-NR8aR8b, R8a represents methyl and R8b} _represents ^{-CH2 - CH2} _{- OCH3 .}

In one embodiment, the present invention comprises compounds of formula (I) and pharma-ceutically acceptable salts and solvates thereof, or any subgroup thereof as described in any of the other embodiments, wherein C _1-6 alkyl in the definition of R ³ , --C _1-6 alkyl-NR ^8a R ^8b , is limited to --CH ₂ --CH ₂ --CH ₂ --.

In one embodiment, the invention comprises a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, or any subgroup thereof, as described in any of the other embodiments, wherein the compound of formula (I) is of formula (Ia) or formula (Ib):

の化合物に限定され、
式中、Ｒ^１ａ、Ｒ^１ｂ、Ｒ^３、Ｒ^４、Ｒ^５ａ、Ｒ^５ｂ、Ｘ^１、Ｘ^２、ｎ１、ｎ２、ｎ３、ｎ４及びハロは、他の実施形態のいずれかに記載の式（Ｉ）の化合物又はそれらの任意のサブグループについて定義される通りである。

Limited to the compounds
wherein R ^1a , R ^1b , R ³ , R ⁴ , R ^5a , R ^5b , X ¹ , X ² , n1, n2, n3, n4 and halo are as defined for the compounds of formula (I) or any subgroup thereof according to any of the other embodiments.

In one embodiment, the compounds of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, or any subgroup thereof described in any of the other embodiments, are limited to compounds of formula (Ia), or a pharma- ceutically acceptable salt or solvate thereof. In one embodiment, the compounds of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, or any subgroup thereof described in any of the other embodiments, are limited to compounds of formula (Ib), or a pharma- ceutically acceptable salt or solvate thereof.

In one embodiment, the present invention comprises a compound of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, or any subgroup thereof, as described in any of the other embodiments, wherein the compound of formula (I) has the formula (I-y):

に限定され、
式中、Ｒ^３は、他の実施形態のいずれかに記載されている式（Ｉ）の化合物又はその任意のサブグループについて定義される通りである。

Limited to
wherein ^R3 is as defined for the compounds of formula (I) or any subgroup thereof as described in any of the other embodiments.

In formula (I-y), n1 is 1, n2 is 2, n3 is 1, and n4 is 1.

In certain embodiments, the compound of formula (I) is compound A:

又はその医薬的に許容される塩若しくは溶媒和物である。

or a pharma- ceutically acceptable salt or solvate thereof.

In certain embodiments, the compound of formula (I) is compound A1:

である。

It is.

In certain embodiments, the compound of formula (I) is compound A2:

である。

It is.

In certain embodiments, the compound of formula (I) is compound A3:

である。

It is.

In certain embodiments, the compound of formula (I) is compound A4-a:

又はその溶媒和物である。

or a solvate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide besylate or a hydrate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate or a solvate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate (compound A4-b) or a hydrate thereof.

In certain embodiments, the menin-MLL inhibitor of formula (I) is compound A4, i.e., crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate.

In certain embodiments, the menin-MLL inhibitor of formula (I) is crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate 0.5-2.0 equivalent hydrate.

In one embodiment, the present invention relates to a subgroup of formula (I) as defined in the general reaction scheme:

In one embodiment, the compound of formula (I) is selected from the group consisting of the exemplified compounds, their tautomers and stereoisomeric forms, and any of their free bases, any pharma- ceutically acceptable salts, and solvates.

In some embodiments, a pharmaceutical composition is provided that includes a pharma- ceutical acceptable carrier and a therapeutically effective amount of any of the combinations described in any of the other embodiments as active ingredients.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and, optionally, at least one other anti-neoplastic agent.

According to an embodiment, the menin-MLL inhibitor is a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof.

In certain embodiments, the menin-MLL inhibitor is compound A or a pharma- ceutically acceptable salt or solvate thereof.

According to certain embodiments, the menin-MLL inhibitor is compound A1.

According to certain embodiments, the menin-MLL inhibitor is compound A2.

According to certain embodiments, the menin-MLL inhibitor is compound A3.

In certain embodiments, the menin-MLL inhibitor is compound A4-a or a solvate thereof.

In certain embodiments, the menin-MLL inhibitor is compound A4-b or a hydrate thereof.

According to certain embodiments, the menin-MLL inhibitor is compound A4.

In certain embodiments, menin-MLL inhibitors may have improved metabolic stability properties.

In certain embodiments, menin-MLL inhibitors may have an extended in vivo half-life (T1/2).

In certain embodiments, menin-MLL inhibitors may have improved oral bioavailability.

In certain embodiments, menin-MLL inhibitors may reduce tumor growth, e.g., in tumors harboring MLL (KMT2A) gene rearrangements/alterations and/or NPM1 mutations.

In certain embodiments, menin-MLL inhibitors may have improved PD properties in vivo over an extended period of time, e.g., inhibition of target gene expression such as MEIS1 and upregulation of differentiation markers over a period of at least 16 hours.

In certain embodiments, menin-MLL inhibitors may have an improved safety profile (e.g., reduced hERG inhibition, improved cardiovascular safety).

In certain embodiments, the menin-MLL inhibitor may be suitable for Q.D. dosing (once daily).

According to embodiments, the BCL-2 inhibitor is selected from obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax, gambogic acid, and venetoclax, or a pharma- ceutically acceptable salt or solvate thereof.

In certain embodiments, the BCL-2 inhibitor is venetoclax, or a pharma- ceutically acceptable salt or solvate thereof.

According to embodiments, the at least one other antineoplastic agent is a hypomethylating agent, a DNA intercalating agent, a pyrimidine analog, a purine analog, a kinase inhibitor, a CD20 inhibitor, an IDH inhibitor, an immunomodulatory antineoplastic agent, or a DHODH inhibitor.

According to embodiments, the at least one other antineoplastic agent is a hypomethylating agent, a DNA intercalating agent, a pyrimidine analog, a purine analog, a kinase inhibitor, a CD20 inhibitor, or an isocitrate dehydrogenase (IDH) inhibitor.

According to embodiments, hypomethylating agents include, but are not limited to, azacitidine, decitabine, or pharma- ceutically acceptable salts or solvates thereof.

According to embodiments, DNA intercalating agents include, but are not limited to, anthracyclines (e.g., daunorubicin, doxorubicin, idarubicin).

According to an embodiment, the DNA intercalating agent is daunorubicin.

According to an embodiment, the DNA intercalating agent is doxorubicin.

According to an embodiment, the DNA intercalating agent is idarubicin.

According to embodiments, pyrimidine analogs include, but are not limited to, cytarabine (ARA-C).

According to an embodiment, the purine analog is fludarabine.

According to an embodiment, the kinase inhibitor is a FLT-3 inhibitor, a BTK inhibitor, an ABL inhibitor, an Aurora inhibitor, or a multikinase inhibitor consisting of two or more of these kinase inhibitors.

According to an embodiment, the kinase inhibitor is a multikinase inhibitor consisting of an FLT-3 inhibitor, an ABL inhibitor, and an Aurora inhibitor. According to an embodiment, such a multikinase inhibitor includes, but is not limited to, KW-2449.

According to an embodiment, the kinase inhibitor is a tyrosine kinase inhibitor.

According to an embodiment, the tyrosine kinase inhibitor is an FLT-3 inhibitor or a BTK inhibitor.

According to embodiments, FLT3 inhibitors include, but are not limited to, sorafenib, sunitinib, midostaurin (PKC412), lestaurtinib (CEP-701), tanzutinib (MLN518), quizartinib (AC220), gilteritinib (ASP2215), and KW-2449.

According to embodiments, BTK inhibitors include, but are not limited to, ibrutinib.

According to embodiments, CD20 inhibitors include, but are not limited to, anti-CD20 antibodies (e.g., obinutuzumab (GA101)).

According to embodiments, IDH inhibitors include, but are not limited to, ivosidenib and enasidenib.

According to embodiments, isocitrate dehydrogenase-1 inhibitors include, but are not limited to, ivosidenib.

According to embodiments, isocitrate dehydrogenase-2 inhibitors include, but are not limited to, enadidenib.

According to embodiments, immunomodulatory anti-neoplastic agents include, but are not limited to, PD-1 inhibitors (e.g., nivolumab, atezolizumab, and pembrolizumab), thalidomide, lenalidomide, pomalidomide, abolished Mycobacterium bovis (BCG), and levamisole.

According to embodiments, PD-1 inhibitors include, but are not limited to, nivolumab, atezolizumab, and pembrolizumab.

According to an embodiment, the DHODH inhibitor has, but is not limited to, the formula (Z):

A compound having the structure:
X is CH or N;
Y is CH or N;
R ¹ is selected from the group consisting of C _1-6 alkyl; C _1-6 alkyl substituted with OH or OCH ₃ ; C _2-6 alkenyl; C _{1-6 haloalkyl; C 1-6 haloalkyl} substituted with OH or OCH ₃ ; C _2-6 haloalkenyl; N(CH ₃ ) ₂ ; C _3-6 cycloalkyl; C _3-6 cycloalkyl substituted with C _1-6 alkyl; and phenyl;
^R2 is

であり、式中、
Ｒ^ａは、Ｃ_１～６アルキル、Ｃ_１～６ハロアルキル、及びＣ_３～６シクロアルキルからなる群から選択され、
Ｒ^ｂは、Ｃ_１～６アルキル、又は、ＯＨ、ハロ、ＣＮ、ＯＣ_１～６アルキル、ＯＣ_１～６ハロアルキル、及びＯＣ_３～６シクロアルキルからなる群から選択されるメンバーで置換されたＣ_１～６アルキルであり、
Ｒ^３は、Ｈ、ハロ、ＣＨ_３及びＯＣＨ_３からなる群から選択され、
Ｒ^４は、
●Ｃ_１～６アルキル、１又は２個のＯＣＨ_３で置換されたＣ_１～６アルキル、Ｃ_３～６シクロアルキル、ＣＨ_３又はＯＣＨ_３で置換されたＣ_３～６シクロアルキル、－ＣＨ_２－Ｃ_３～６シクロアルキル、及び

where:
R ^a is selected from the group consisting of C _1-6 alkyl, C _1-6 haloalkyl, and C _3-6 cycloalkyl;
R ^b is C _1-6 alkyl or C 1-6 alkyl substituted with a member selected from the group consisting of OH, halo, CN, OC _1-6 alkyl, OC _1-6 haloalkyl _, and OC _3-6 cycloalkyl;
^R3 is selected from the group consisting of H, halo, _CH3 and _OCH3 ;
^R4 is
- _{C1-6 alkyl, C1-6 alkyl} substituted with 1 or 2 OCH3, _C3-6 cycloalkyl, _C3-6 cycloalkyl substituted with _CH3 or _OCH3 , _{-CH2 -C3-6 cycloalkyl} , and

●

●

●

●

並びに、
●

and,
●

からなる群から選択され、
式中、各Ｒ^ｃは、独立して、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＮＯ_２；ＯＨ；Ｏ－ＣＨ_２ＣＨ_２ＯＨ、及びＯＣ_１～６アルキル、からなる群から選択され、
Ｒ^ｄは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＣＮ、並びに、ＯＣ_１～６アルキルからなる群から選択され、
Ｒ^ｇは、Ｈ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル、並びに、ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル、からなる群から選択され、
ｎは、１又は２である］、
又はその医薬的に許容される塩、同位体、Ｎ－オキシド、溶媒和物、若しくは立体異性体、
あるいは、

is selected from the group consisting of
wherein each R ^c is independently selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; NO ₂ ; OH; O—CH ₂ CH ₂ OH, and OC _1-6 alkyl;
R ^d is selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; CN, and OC _1-6 alkyl;
R ^g is selected from the group consisting of H; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _{1-6 haloalkyl, and C 1-6 haloalkyl} substituted with a member selected from the group consisting of OH and OCH ₃ ;
n is 1 or 2;
or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof;
or,

or a pharma- ceutically acceptable salt, N-oxide, solvate, or stereoisomer thereof.

According to an embodiment, the DHODH inhibitor has, but is not limited to, the formula (Z):

A compound having the structure:
X is CH or N;
Y is CH or N;
R ¹ is selected from the group consisting of C _1-6 alkyl; C _1-6 alkyl substituted with OH or OCH ₃ ; C _2-6 alkenyl; C _{1-6 haloalkyl; C 1-6 haloalkyl} substituted with OH or OCH ₃ ; C _2-6 haloalkenyl; N(CH ₃ ) ₂ ; C _3-6 cycloalkyl; C _3-6 cycloalkyl substituted with C _1-6 alkyl; and phenyl;
^R2 is

であり、式中、
Ｒ^ａは、Ｃ_１～６アルキル、Ｃ_１～６ハロアルキル、及びＣ_３～６シクロアルキルからなる群から選択され、
Ｒ^ｂは、Ｃ_１～６アルキル、又は、ＯＨ、ハロ、ＣＮ、ＯＣ_１～６アルキル、ＯＣ_１～６ハロアルキル、及びＯＣ_３～６シクロアルキルからなる群から選択されるメンバーで置換されたＣ_１～６アルキルであり、
Ｒ^３は、Ｈ、ハロ、ＣＨ_３及びＯＣＨ_３からなる群から選択され、
Ｒ^４は、
●Ｃ_１～６アルキル、１又は２個のＯＣＨ_３で置換されたＣ_１～６アルキル、Ｃ_３～６シクロアルキル、ＣＨ_３又はＯＣＨ_３で置換されたＣ_３～６シクロアルキル、－ＣＨ_２－Ｃ_３～６シクロアルキル、及び

where:
R ^a is selected from the group consisting of C _1-6 alkyl, C _1-6 haloalkyl, and C _3-6 cycloalkyl;
R ^b is C _1-6 alkyl or C 1-6 alkyl substituted with a member selected from the group consisting of OH, halo, CN, OC _1-6 alkyl, OC _1-6 haloalkyl _, and OC _3-6 cycloalkyl;
^R3 is selected from the group consisting of H, halo, _CH3 and _OCH3 ;
^R4 is
- _{C1-6 alkyl, C1-6 alkyl} substituted with 1 or 2 OCH3, _C3-6 cycloalkyl, _C3-6 cycloalkyl substituted with _CH3 or _OCH3 , _{-CH2 -C3-6 cycloalkyl} , and

●

●

●

●

並びに、
●

and,
●

からなる群から選択され、
式中、各Ｒ^ｃは、独立して、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＮＯ_２；ＯＨ；Ｏ－ＣＨ_２ＣＨ_２ＯＨ、及びＯＣ_１～６アルキル、からなる群から選択され、
Ｒ^ｄは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＣＮ、並びに、ＯＣ_１～６アルキルからなる群から選択され、
Ｒ^ｇは、Ｈ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル、並びに、ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル、からなる群から選択され、
ｎは、１又は２である］、
又はその医薬的に許容される塩、同位体、Ｎ－オキシド、溶媒和物、若しくは立体異性体を含む。

is selected from the group consisting of
wherein each R ^c is independently selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; NO ₂ ; OH; O—CH ₂ CH ₂ OH, and OC _1-6 alkyl;
R ^d is selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; CN, and OC _1-6 alkyl;
R ^g is selected from the group consisting of H; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _{1-6 haloalkyl, and C 1-6 haloalkyl} substituted with a member selected from the group consisting of OH and OCH ₃ ;
n is 1 or 2;
or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In relation to formula (Z), the following definitions apply:

The term "alkenyl" includes unsaturated aliphatic groups similar in length and possible substitution to alkyl, except that they contain at least one double bond. For example, the term "alkenyl" includes straight chain alkenyl groups such as ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, and the like. The term alkenyl further includes alkenyl groups which include oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more carbons of the hydrocarbon backbone. In certain embodiments, straight chain or branched chain alkenyl groups have 6 or fewer carbon atoms in their backbone (e.g., _C2-6 for straight chain, _C3-6 for branched chain).

The term "haloalkyl" refers to a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms in the chain, optionally having hydrogen replaced with halogen. The term "C _1-6 haloalkyl" as used herein refers to a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms in the chain, optionally having hydrogen replaced with halogen. The term "C _1-4 haloalkyl" as used herein refers to a straight-chain or branched-chain alkyl group having 1 to 4 carbon atoms in the chain, optionally having hydrogen replaced with halogen. Examples of "haloalkyl" groups include trifluoromethyl ( _CF3 ), difluoromethyl ( _{CF2H), monofluoromethyl (CH2F} ), _{pentafluoroethyl} ( _CF2CF3 ), tetrafluoroethyl ( _CHFCF3 ), monofluoroethyl ( _CH2CH2F ), trifluoroethyl ( _CH2CF3 ), tetrafluorotrifluoromethylethyl (CF _{( CF3 ) 2} ), as well as groups _which would be considered equivalent to any one of the foregoing examples given the ordinary skill in the art and the teachings provided herein.

The term "haloalkenyl" includes unsaturated aliphatic groups similar in length and optional substitution to the alkyls described above, but containing at least one double bond and optionally having hydrogens replaced with halogens, and having 1 to 6 carbon atoms in the chain.

The term "aryl" refers to a monocyclic aromatic carbocycle (a ring structure in which the ring atoms are all carbon) having six atoms per ring. (The carbon atoms in an aryl group are sp2 hybridized.)

The term "heteroaryl" refers to a monocyclic or fused bicyclic heterocycle (a ring structure having ring atoms selected from carbon atoms and up to four heteroatoms selected from nitrogen, oxygen, and sulfur) having 3 to 9 ring atoms per heterocycle. Illustrative examples of heteroaryl groups include the following entities, in the form of appropriately bonded moieties:

Those of skill in the art will recognize that the species listed or exemplified above are not all inclusive and that additional species may be selected within the scope of these defined terms.

The term "variable point of attachment" means that a group may be attached at two or more alternative positions within a structure. The bond always replaces a hydrogen atom on one of the ring atoms. In other words, all permutations of the bonds, as shown in the diagram below, are represented by a single diagram.

In one embodiment of the present invention, the DHODH inhibitor is a compound of formula (Z), where X is CH.

In one embodiment of the present invention, the DHODH inhibitor is a compound of formula (Z), where X is N.

In one embodiment of the present invention, the DHODH inhibitor is a compound of formula (Z), where Y is CH.

In one embodiment of the present invention, the DHODH inhibitor is a compound of formula (Z), where Y is N.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), where R ¹ is C _{1-4 alkyl; C 1-4} alkyl substituted with OH or OCH ₃ ; C _2-4 alkenyl; C _{1-4 haloalkyl; C 1-4 haloalkyl} substituted with OH _or OCH ₃ ; C _2-4 haloalkenyl; N(CH ₃ ) ₂ ; cyclopropyl, cyclopropyl substituted with C _1-4 alkyl; or phenyl.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein R ¹ is CH ₃ , CH ₂ CH ₃ ,

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein R ¹ is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R2 is

であり、
式中、Ｒ^ｂは、ＯＨ、ハロ、ＣＮ、ＯＣ_１～４アルキル、ＯＣ_１～４ハロアルキル、若しくはＯＣ_３～６シクロアルキルで置換されたＣ_１～４アルキルであり、
Ｒ^ａが、Ｃ_１～４アルキル、Ｃ_１～４ハロアルキル、又はＣ_３～６シクロアルキルである。

and
wherein R ^b is C _1-4 alkyl substituted with OH, halo, CN, OC _1-4 alkyl, OC _1-4 haloalkyl, or OC _3-6 cycloalkyl;
R ^a is C _1-4 alkyl, C _1-4 haloalkyl, or C _3-6 cycloalkyl.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R2 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein R ³ is H.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein R ³ is F.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), where ^R3 is _CH3 .

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), where ^R3 is _OCH3 .

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、式中、
各Ｒ^ｃは、独立して、Ｈ；ハロ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～アルキル；Ｃ_１～４ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～４ハロアルキル、並びに、ＮＯ_２、からなる群から選択され、
Ｒ^ｄは、Ｈ；ハロ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、若しくはＯＣＦ_３で置換されたＣ_１～４アルキル；Ｃ_１～４ハロアルキル；ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～４ハロアルキル；又はＯＣ_１～４アルキル；ＣＮ、並びに、ＯＣ_１～６アルキル、からなる群から選択され、
ｎは、１又は２である。

where:
each R ^c is independently selected from the group consisting of H; halo; C _1-4 alkyl; C _1- alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-4 haloalkyl; C _1-4 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ , and NO ₂ ;
R ^d is selected from the group consisting of H; halo; C _1-4 alkyl; C _1-4 alkyl substituted with OH, OCH ₃ , SCH ₃ , or OCF ₃ ; C _1-4 haloalkyl; C _1-4 haloalkyl substituted with OH or OCH ₃ ; or OC _1-4 alkyl; CN, and OC _1-6 alkyl;
n is 1 or 2.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、
各Ｒ^ｃは、Ｈ、ハロ、Ｃ_１～４アルキル、Ｃ_１～４ハロアルキル、ＮＯ_２、Ｏ－ＣＨ_２ＣＨ_２ＯＨ、及びＯＣ_１～４アルキルからなる群から独立して選択され、
Ｒ^ｄは、Ｈ、ハロ、Ｃ_１～４アルキル、ＣＮ、及びＯＣ_１～６アルキルからなる群から選択され、
ｎは、１又は２である。

and
each R ^c is independently selected from the group consisting of H, halo, C _1-4 alkyl, C _1-4 haloalkyl, NO ₂ , O—CH ₂ CH ₂ OH, and OC _1-4 alkyl;
R ^d is selected from the group consisting of H, halo, C _1-4 alkyl, CN, and OC _1-6 alkyl;
n is 1 or 2.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、
式中、Ｒ^ｃは、Ｈ；ハロ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～アルキル；Ｃ_１～４ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されているＣ_１～４ハロアルキル、からなる群から独立して選択され、Ｒ^ｄは、ハロ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、若しくはＯＣＦ_３で置換されたＣ_１～４アルキル；Ｃ_１～４ハロアルキル；ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～４ハロアルキル；又はＯＣ_１～４アルキル；ＣＮ、並びに、ＯＣ_１～６アルキル、からなる群から選択される。

and
wherein R ^c is independently selected from the group consisting of H; halo; C _1-4 alkyl; C _1- alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-4 haloalkyl; C _1-4 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; and R ^d is selected from the group consisting of halo; C _1-4 alkyl; C _1-4 alkyl substituted with OH, OCH ₃ , SCH ₃ , or OCF ₃ ; C _1-4 haloalkyl; C _1-4 haloalkyl substituted with OH or OCH ₃ ; or OC _1-4 alkyl; CN, and OC _1-6 alkyl.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、式中、
各Ｒ^ｃは、Ｈ、ハロ、Ｃ_１～４アルキル、Ｃ_１～４ハロアルキル、ＯＣ_１～４アルキル、及びＯＨからなる群から独立して選択され、
Ｒ^ｄは、ハロ、Ｃ_１～４アルキル及びＯＣ_１～４アルキルからなる群から選択され、
ｎは、１又は２である。

where:
each R ^c is independently selected from the group consisting of H, halo, C _1-4 alkyl, C _1-4 haloalkyl, OC _1-4 alkyl, and OH;
R ^d is selected from the group consisting of halo, C _1-4 alkyl, and OC _1-4 alkyl;
n is 1 or 2.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、式中、
Ｒ^ｃは、Ｈ；ハロ；Ｃ_１～４アルキル、ＯＨ、ＯＣＨ_３、ＳＣＨ_３、若しくはＯＣＦ_３で置換されたＣ_１～４アルキル；Ｃ_１～４ハロアルキル；ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～４ハロアルキル；又はＯＣ_１～４アルキル；
Ｒ^ｄは、ハロ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、若しくはＯＣＦ_３で置換されたＣ_１～４アルキル；Ｃ_１～４ハロアルキル；又はＯＨ若しくはＯＣＨ_３で置換されたＣ_１～４ハロアルキルであり、
Ｒ^ｇは、Ｈ；Ｃ_１～４アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、若しくはＯＣＦ_３で置換されたＣ_１～４アルキル；Ｃ_１～４ハロアルキル；又は、ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～４ハロアルキルである。

where:
R ^c is H; halo; C _{1-4 alkyl, C 1-4} alkyl substituted with OH, OCH ₃ , SCH ₃ , or OCF _{3 ;} C _1-4 haloalkyl; C _1-4 haloalkyl substituted with OH or OCH ₃ ; or OC _1-4 alkyl;
R ^d is halo; C _1-4 alkyl; C _1-4 alkyl substituted with OH, OCH ₃ , SCH ₃ , or OCF ₃ ; C _1-4 haloalkyl; or C _1-4 haloalkyl substituted with OH or OCH ₃ ;
R ^g is H; C _1-4 alkyl; C _1-4 alkyl substituted with OH, OCH ₃ , SCH ₃ , or OCF ₃ ; C _1-4 haloalkyl; or C _1-4 haloalkyl substituted with OH or OCH ₃ .

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein:
^R4 is

であり、式中、
Ｒ^ｃは、Ｈ又はハロであり、
Ｒ^ｄは、Ｃ_１～４アルキルであり、
Ｒ^ｇは、Ｈである。

where:
R ^c is H or halo;
R ^d is C _1-4 alkyl;
^Rg is H.

In one embodiment of the invention, the DHODH inhibitor is a compound of formula (Z), wherein ^R4 is

である。

It is.

In one embodiment of the invention, the DHODH inhibitor is
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-phenylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(3,3,3-trifluoroprop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,6-dichlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
2-(2,6-dichlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-4-cyclopropyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one;
2-(6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-1-oxo-4-(prop-1-en-2-yl)isoquinolin-2(1H)-yl)benzonitrile;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylphthalazin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-isopropylphthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylphthalazin-1(2H)-one;
4-ethyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)phthalazin-1(2H)-one;
4-ethyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(o-tolyl)phthalazin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-4-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-(2-hydroxypropan-2-yl)isoquinolin-1(2H)-one;
4-(dimethylamino)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(5-chloro-3-methyl-1H-pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-(prop-1-en-2-yl)-6-(o-tolyl)-1,6-naphthyridin-5(6H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-methyl-6-(o-tolyl)pyrido[2,3-d]pyridazin-5(6H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-tolyl)pyrido[2,3-d]pyridazin-5(6H)-one;
6-(2-chloro-6-fluorophenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-(prop-1-en-2-yl)-1,6-naphthyridin-5(6H)-one;
6-(2-chloro-6-fluorophenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;
(S)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(o-tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6H)-one;
(R)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(o-tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylthiazol-5-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-tolyl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methoxyphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
racemic-4-(sec-butyl)-2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoroisoquinolin-1(2H)-one;
2-(3-chloro-6-methoxypyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-4-methylpyridin-3-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluoro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylphthalazin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
(S ^* )-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
(R ^* )-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxy-4-methylpyridin-3-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(5-chloro-3-methyl-1H-pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(3-chloro-2-methoxy-5-methylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-2-methoxy-5-methylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(4-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-3-(2-hydroxyethoxy)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluorophenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-difluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-6-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxy-3,5-dimethylpyridin-4-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,5-difluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-6-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-dimethylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(4-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluorophenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-3,5-dimethylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-methyl-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluoro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(3,3,3-trifluoroprop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxyphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methoxypyridin-4-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,5-dimethylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(2-ethylphenyl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxypyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methoxypyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(methyl-d ₃ )phenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylpyridin-5-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxyphenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-6-methoxypyridin-2-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyrazin-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(2-fluoro-5-methylphenyl)-8-isopropyl-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro 4-isopropyl-2-(4-methylpyridazin-3-yl)isoquinolin-1(2H)-one;
(S)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
(R)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(5-methylpyridin-4-yl)isoquinolin-1(2H)-one;
2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-2-methoxypyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-cyclohexyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-6-methylpyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-cyclopentyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-4-methoxypyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R,2S)-2-methylcyclohexyl)isoquinolin-1(2H)-one;
2-(1,3-dimethoxypropan-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-5-methylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S,2R)-2-methylcyclohexyl)isoquinolin-1(2H)-one;
2-(cyclopropylmethyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(1-methoxybutan-2-yl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyridin-2-yl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((cis)-3-methoxycyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2R ^* )-2-methylcyclohexyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2S ^* )-2-methylcyclohexyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(pentan-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2R ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2S ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2S ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2R ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((cis)-3-methoxycyclohexyl)isoquinolin-1(2H)-one;
2-(bicyclo[2.2.1]heptan-1-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-3-methylpyridin-4-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(2-methoxyphenyl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylisothiazol-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(5-methylisothiazol-4-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(2-(trifluoromethyl)phenyl)-1,6-naphthyridin-5(6H)-one;
2-(3,6-dimethylpyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-dimethylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylpyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methylpyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-hydroxy-5-methylpyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(2-(difluoromethyl)phenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;
Compounds of formula (Z) selected from the group consisting of 6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-hydroxy-3-methylpyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one; and 2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-D ₃ -tolyl)-1,6-naphthyridin-5(6H)-one,
and optionally one or more of a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, and stereoisomer thereof.

In one embodiment of the invention, the DHODH inhibitor is
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-phenylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(3,3,3-trifluoroprop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,6-dichlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
2-(2,6-dichlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-4-cyclopropyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one;
2-(6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-1-oxo-4-(prop-1-en-2-yl)isoquinolin-2(1H)-yl)benzonitrile;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylphthalazin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-isopropylphthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-isopropylphthalazin-1(2H)-one;
4-ethyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)phthalazin-1(2H)-one;
4-ethyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(o-tolyl)phthalazin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-4-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(3-fluorophenyl)-4-(2-hydroxypropan-2-yl)isoquinolin-1(2H)-one;
4-(dimethylamino)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(5-chloro-3-methyl-1H-pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-(prop-1-en-2-yl)-6-(o-tolyl)-1,6-naphthyridin-5(6H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-methyl-6-(o-tolyl)pyrido[2,3-d]pyridazin-5(6H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-tolyl)pyrido[2,3-d]pyridazin-5(6H)-one;
6-(2-chloro-6-fluorophenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-(prop-1-en-2-yl)-1,6-naphthyridin-5(6H)-one;
6-(2-chloro-6-fluorophenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;
(S)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(o-tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6H)-one;
(R)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(o-tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylthiazol-5-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-tolyl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methoxyphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
racemic-4-(sec-butyl)-2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoroisoquinolin-1(2H)-one;
2-(3-chloro-6-methoxypyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-4-methylpyridin-3-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluoro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylphthalazin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
(S ^* )-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
(R ^* )-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxy-4-methylpyridin-3-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(5-chloro-3-methyl-1H-pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(3-chloro-2-methoxy-5-methylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-2-methoxy-5-methylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-5-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(4-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-3-(2-hydroxyethoxy)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluorophenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-difluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-6-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxy-3,5-dimethylpyridin-4-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,5-difluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluoro-6-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-2-methylphenyl)-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-dimethylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(4-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-fluorophenyl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-3,5-dimethylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-methyl-4-(prop-1-en-2-yl)-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluoro-3-methoxyphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(3,3,3-trifluoroprop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-methoxyphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methoxypyridin-4-yl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-2-methylphenyl)-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
2-(2,5-dimethylphenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(2-ethylphenyl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxypyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(5-fluoro-2-methoxypyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(methyl-d ₃ )phenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylpyridin-5-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxyphenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(3-fluoro-6-methoxypyridin-2-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyrazin-2-yl)isoquinolin-1(2H)-one;
2-(2-chloro-5-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-6-(2-fluoro-5-methylphenyl)-8-isopropyl-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro 4-isopropyl-2-(4-methylpyridazin-3-yl)isoquinolin-1(2H)-one;
(S)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
(R)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(5-methylpyridin-4-yl)isoquinolin-1(2H)-one;
2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-2-methoxypyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-cyclohexyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-6-methylpyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-cyclopentyl-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(3-chloro-4-methoxypyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R,2S)-2-methylcyclohexyl)isoquinolin-1(2H)-one;
2-(1,3-dimethoxypropan-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-5-methylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S,2R)-2-methylcyclohexyl)isoquinolin-1(2H)-one;
2-(cyclopropylmethyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(1-methoxybutan-2-yl)isoquinolin-1(2H)-one;
2-(2-chlorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyridin-2-yl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((cis)-3-methoxycyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2R ^* )-2-methylcyclohexyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2S ^* )-2-methylcyclohexyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(pentan-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2R ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2S ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1R ^* ,2S ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((1S ^* ,2R ^* )-2-methylcyclopentyl)isoquinolin-1(2H)-one;
racemic-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-((cis)-3-methoxycyclohexyl)isoquinolin-1(2H)-one;
2-(bicyclo[2.2.1]heptan-1-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methoxy-3-methylpyridin-4-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(2-methoxyphenyl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylisothiazol-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(5-methylisothiazol-4-yl)isoquinolin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(2-(trifluoromethyl)phenyl)-1,6-naphthyridin-5(6H)-one;
2-(3,6-dimethylpyridin-2-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(2,5-dimethylpyridin-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(4-methylpyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(3-methylpyridin-4-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-methylpyridin-3-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-hydroxy-5-methylpyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one;
6-(2-(difluoromethyl)phenyl)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;
Compounds of formula (Z) selected from the group consisting of 6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(2-hydroxy-3-methylpyridin-4-yl)-4-isopropylisoquinolin-1(2H)-one; and 2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-D ₃ -tolyl)-1,6-naphthyridin-5(6H)-one,
and optionally one or more of its pharma- ceutically acceptable salts, isotopes, N-oxides, solvates, and stereoisomers, or 2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-(2-fluoro-4-nitrophenyl)-4-iodoisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-7-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-6-fluoro-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-7-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-6-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
or a pharma- ceutically acceptable salt, N-oxide, solvate, or stereoisomer thereof.

In one embodiment of the invention, the DHODH inhibitor is
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one;
2-(2-chloro-4-methylpyridin-3-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-4-(1-methylcyclopropyl)isoquinolin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;
2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-3-fluoro-8-isopropyl-6-(o-tolyl)-1,6-naphthyridin-5(6H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)phthalazin-1(2H)-one;
2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylphthalazin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(methyl-d3)phenyl)isoquinolin-1(2H)-one;
(R)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-2-(o-tolyl)-4-(1,1,1-trifluoropropan-2-yl)isoquinolin-1(2H)-one;
6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one;
2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;
and optionally one or more of a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, and stereoisomer thereof.

In one embodiment of the present invention, the DHODH inhibitor has the formula (Za):

を有する式（Ｚ）の化合物［式中、
Ｙは、ＣＨ又はＮであり、
Ｒ^１は、Ｃ_１～６アルキル；ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～６アルキル；Ｃ_２～６アルケニル；Ｃ_１～６ハロアルキル；ＯＨ若しくはＯＣＨ_３で置換されたＣ_１～６ハロアルキル；Ｃ_２～６ハロアルケニル；Ｎ（ＣＨ_３）_２；Ｃ_３～６シクロアルキル；Ｃ_１～６アルキルで置換されたＣ_３～６シクロアルキル；及びフェニルからなる群から選択され、
Ｒ^２は、

A compound of formula (Z) having the formula
Y is CH or N;
R ¹ is selected from the group consisting of C _1-6 alkyl; C _1-6 alkyl substituted with OH or OCH ₃ ; C _2-6 alkenyl; C _{1-6 haloalkyl; C 1-6 haloalkyl} substituted with OH or OCH ₃ ; C _2-6 haloalkenyl; N(CH ₃ ) ₂ ; C _3-6 cycloalkyl; C _3-6 cycloalkyl substituted with C _1-6 alkyl; and phenyl;
^R2 is

であり、
Ｒ^３は、Ｈ、ハロ、ＣＨ_３及びＯＣＨ_３からなる群から選択され、
Ｒ^４は、
Ｃ_１～６アルキル；１又は２個のＯＣＨ_３で置換されたＣ_１～６アルキル；Ｃ_３～６シクロアルキル；ＣＨ_３又はＯＣＨ_３で置換されたＣ_３～６シクロアルキル；－ＣＨ_２－Ｃ_３～６シクロアルキル、及び

and
^R3 is selected from the group consisting of H, halo, _CH3 and _OCH3 ;
^R4 is
C _1-6 alkyl; C _1-6 alkyl substituted with 1 or 2 OCH ₃ ; C _3-6 cycloalkyl; C _3-6 cycloalkyl substituted with CH ₃ or OCH ₃ ; -CH ₂ -C _3-6 cycloalkyl, and

並びに、

and,

から選択され、
式中、Ｒ^ｃは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＮＯ_２；ＯＨ；Ｏ－ＣＨ_２ＣＨ_２ＯＨ、及びＯＣ_１～６アルキル、からなる群から独立して選択され、
Ｒ^ｄは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＣＮ、並びに、ＯＣ_１～６アルキルからなる群から選択され、
Ｒ^ｇは、Ｈ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル、並びに、ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル、からなる群から選択され、
ｎは、１又は２である］、
又はその医薬的に許容される塩、溶媒和物、立体異性体、同位体バリアント、若しくはＮ－オキシドである。

is selected from
wherein R ^c is independently selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; NO ₂ ; OH; O—CH ₂ CH ₂ OH, and OC _1-6 alkyl;
R ^d is selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; CN, and OC _1-6 alkyl;
R ^g is selected from the group consisting of H; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _{1-6 haloalkyl, and C 1-6 haloalkyl} substituted with a member selected from the group consisting of OH and OCH ₃ ;
n is 1 or 2;
or a pharma- ceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof.

In one embodiment of the present invention, the DHODH inhibitor has the formula (Zb):

を有する式（Ｚ）の化合物［式中、
Ｙは、ＣＨ又はＮであり、
Ｒ^１は、Ｃ_１～６アルキル、Ｃ_１～６ハロアルキル及びＣ_２～６アルケニルからなる群から選択され、
Ｒ^２は、

A compound of formula (Z) having the formula
Y is CH or N;
R ¹ is selected from the group consisting of C _1-6 alkyl, C _1-6 haloalkyl, and C _2-6 alkenyl;
^R2 is

であり、
Ｒ^３は、Ｈ、ハロ、及びＯＣＨ_３からなる群から選択され、
Ｒ^４は、

and
^R3 is selected from the group consisting of H, halo, and _OCH3 ;
^R4 is

からなる群から選択され、
式中、Ｒ^ｃは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル、並びに、ＮＯ_２からなる群から選択され、
Ｒ^ｄは、Ｈ；ハロ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル；ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル；ＣＮ、並びに、ＯＣ_１～６アルキルからなる群から選択され、
Ｒ^ｇは、Ｈ；Ｃ_１～６アルキル；ＯＨ、ＯＣＨ_３、ＳＣＨ_３、及びＯＣＦ_３からなる群から選択されるメンバーで置換されたＣ_１～６アルキル；Ｃ_１～６ハロアルキル、並びに、ＯＨ及びＯＣＨ_３からなる群から選択されるメンバーで置換されたＣ_１～６ハロアルキル、からなる群から選択され、
ｎは、１である］、
又はその医薬的に許容される塩、溶媒和物、立体異性体、同位体バリアント、若しくはＮ－オキシドである。

is selected from the group consisting of
wherein R ^c is selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ , and NO ₂ ;
R ^d is selected from the group consisting of H; halo; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _1-6 haloalkyl; C _1-6 haloalkyl substituted with a member selected from the group consisting of OH and OCH ₃ ; CN, and OC _1-6 alkyl;
R ^g is selected from the group consisting of H; C _1-6 alkyl; C _1-6 alkyl substituted with a member selected from the group consisting of OH, OCH ₃ , SCH ₃ , and OCF ₃ ; C _{1-6 haloalkyl, and C 1-6 haloalkyl} substituted with a member selected from the group consisting of OH and OCH ₃ ;
n is 1;
or a pharma- ceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof.

Exemplary compounds of formula (Z) useful in the methods of the invention are described below with reference to exemplary synthetic schemes for their general preparation and the following examples. To obtain the various compounds herein, one skilled in the art will appreciate that starting materials can be suitably selected such that the ultimately desired substituents are retained throughout the reaction scheme, with or without appropriate protection, to obtain the desired product. Alternatively, it may be necessary or desirable to use, in place of the ultimately desired substituent, a suitable group that is retained throughout the reaction scheme and can be appropriately substituted with the desired substituent. Unless otherwise specified, the variables are as defined above with reference to formula (Z). The reaction can be carried out between the melting point and the reflux temperature of the solvent, preferably between 0° C. and the reflux temperature of the solvent. The reaction can be heated using conventional or microwave heating. The reaction can also be carried out in a closed pressure vessel at a temperature higher than the normal reflux temperature of the solvent.

All abbreviations used in the general scheme and examples of formula (Z) are as defined in Table 1A. Variables are as defined within ranges or as specifically defined in the general scheme.

PREPARATIVE EXAMPLES Exemplary compounds of formula (Z) useful in the methods of the present invention will now be described by reference to the following exemplary synthetic schemes for their general preparation and the following specific examples.

Scheme 1

スキーム１に従って、エチル２－（ベンジルオキシ）アセテートから３工程で式（ＩＩ）の１，２，４－トリアゾール－５（４Ｈ）－オン化合物（式中、ＰＧはＢｎである）を調製する。第１の工程では、ＥｔＯＨなどの好適な溶媒中において、エチル２－（ベンジルオキシ）アセテートをヒドラジン水和物と反応させることによって、２－（ベンジルオキシ）アセトヒドラジドを、７０～８５℃の範囲の温度で調製する。水などの適切な溶媒中でのヒドラジドと式Ｒ^ａ－ＮＣＯ（式中、Ｒ^ａはＣ_１～６アルキルである）のイソシアネートとの反応；対応するセミカルバジドを提供する。その後、水などの好適な溶媒中で、ＮａＯＨなどの好適な塩基を用いてセミカルバジドを環化することにより、式（ＩＩ）の化合物（式中、ＰＧはＢｎである）を提供する。

According to Scheme 1, 1,2,4-triazol-5(4H)-one compounds of formula (II), where PG is Bn, are prepared in three steps from ethyl 2-(benzyloxy)acetate. In the first step, 2-(benzyloxy)acetohydrazide is prepared by reacting ethyl 2-(benzyloxy)acetate with hydrazine hydrate in a suitable solvent such as EtOH at a temperature ranging from 70-85° C. Reaction of the hydrazide with an isocyanate of formula R ^a -NCO, where R ^a is C _1-6 alkyl, in a suitable solvent such as water; provides the corresponding semicarbazide. Subsequent cyclization of the semicarbazide with a suitable base such as NaOH in a suitable solvent such as water provides compounds of formula (II), where PG is Bn.

Compounds of formula (II) where R ^a is C _1-6 haloalkyl or C _3-6 cycloalkyl can be prepared as described above using an appropriately substituted compound of formula R ^a -NCO where R ^a is C _1-6 haloalkyl or C _3-6 cycloalkyl.

Protecting group exchange from a compound of formula (II) where PG is Bn to a compound of formula (II) where PG is TBDPS is accomplished in two steps using established methods such as those described in T. W. Greene and P. G. M. Wuts, "Protective Groups in Organic Synthesis," 3 ed., John Wiley & Sons, 1999. In the first step, deprotection of the benzyl group is accomplished under hydrogenolysis conditions well known to those skilled in the art to provide the alcohol. For example, deprotection is accomplished using a palladium catalyst such as Pd/C under _H2 in a suitable solvent such as EtOH, MeOH, EtOAc, or mixtures thereof, preferably EtOH, with or without HCl for 4-72 hours. In the second step, compounds of formula (II) where PG is TBDPS are obtained using a suitable base such as tert-butyldiphenylsilyl chloride, imidazole, dimethylaminopyridine, pyridine, in a solvent such as DMF, DCM, at temperatures ranging from 0° C. to room temperature.

Scheme 2

スキーム２に従って、Ｒ^３がＨである式（ＸＩＶ）の化合物を、Ｎ－ヨードスクシンイミド（ＮＩＳ）などのハロゲン化試薬で処理し、アセトニトリルなどの非プロトン性溶媒中、加熱条件下で処理して、式（ＩＩＩ）のハロゲン化化合物（式中、ＨＡＬはヨウドである）を得る。式Ｒ^１－Ｂ（ＯＨ）_２の化合物を当業者に周知の鈴木カップリング条件下で式（ＩＩＩ）の化合物と反応させて、式（ＩＶ）の化合物を提供する。例えば、ＨＡＬがヨウ化物である式（ＩＩＩ）の化合物を、Ｒ^１－Ｂ（ＯＨ）_２（式中、Ｒ^１は、式（Ｚ）を参照して本明細書で定義されている、任意選択的に置換されていているＣ_２～６アルケニル又はアリールである）などの市販の又は合成により入手可能なボロン酸（又はボロン酸エステル）、ビス（トリフェニルホスフィン）パラジウム（ＩＩ）ジクロリド、テトラキス（トリフェニルホスフィン）パラジウムなどのパラジウム触媒、リン酸カリウム、Ｃｓ_２ＣＯ_３などの適切な塩基と、ジオキサン、水、エタノール、又はこれらの混合物などの好適な溶媒中で反応させて、式化合物（ＩＶ）の化合物を得る。Ｒ^３がＨである式（ＩＶ）の化合物を、式Ｒ^４－Ｂ（ＯＨ）_２の化合物と、当業者に公知の銅（ＩＩ）媒介Ｃｈａｎ－Ｌａｍカップリング条件下で反応させて、式（Ｖ）の化合物を得る（式中、ＨＡＬは臭化物であり、ＸはＣＨであり、Ｒ^３はＨである）。例えば、式（ＩＶ）の化合物を、式Ｒ^４－Ｂ（ＯＨ）_２（式中、Ｒ^４は式（Ｚ）を参照して本明細書で定義される通りである）の化合物、酢酸銅（ＩＩ）などの触媒、ピリジン、ＮＥｔ_３などの塩基と、ＤＣＭ、ＡＣＮ、ジオキサン、ＴＨＦなどの好適な溶媒中で反応させて、式（Ｖ）の化合物を得る。

According to Scheme 2, a compound of formula (XIV) where R ³ is H is treated with a halogenating reagent such as N-iodosuccinimide (NIS) in an aprotic solvent such as acetonitrile under heating conditions to give a halogenated compound of formula (III) where HAL is iodo. A compound of formula R ¹ -B(OH) ₂ is reacted with a compound of formula (III) under Suzuki coupling conditions well known to those skilled in the art to provide a compound of formula (IV). For example, a compound of formula (III) where HAL is an iodide is reacted with a commercially available or synthetically available boronic acid (or boronic acid ester) such as R ¹ -B(OH) ₂ (where R ¹ is an optionally substituted C _2-6 alkenyl or aryl as defined herein with reference to formula (Z)), a palladium catalyst such as bis(triphenylphosphine)palladium(II) dichloride, tetrakis(triphenylphosphine)palladium, a suitable base such as potassium phosphate, Cs ₂ CO ₃ , in a suitable solvent such as dioxane, water, ethanol, or a mixture thereof to give a compound of formula (IV). A compound of formula (IV) where R ³ is H is reacted with a compound of formula R ⁴ -B(OH) ₂ under copper(II) mediated Chan-Lam coupling conditions known to those skilled in the art to give a compound of formula (V) (where HAL is a bromide, X is CH, and R ³ is H). For example, a compound of formula (IV) is reacted with a compound of formula R ⁴ -B(OH) ₂ (wherein R ⁴ is as defined herein with reference to formula (Z)), a catalyst such as copper(II) acetate, a base such as pyridine, NEt ₃ , in a suitable solvent such as DCM, ACN, dioxane, THF, to give a compound of formula (V).

Scheme 3

スキーム３に従って、式（Ｖ）の化合物（式中、Ｒ^１は任意選択的に置換されたＣ_２～６アルケニルであり、Ｒ^３はＨであり、Ｒ^４は式（Ｚ）を参照して本明細書に記載の適切に置換されたフェニルであり、ＨＡＬはＢｒである）を、式（ＩＩ）の市販の又は合成により入手可能な求核性化合物（式中、Ｒ^ａはＣ_１～６アルキルである）、例えば、好適に保護されたトリアゾロン（式中、ＰＧは、ベンジル、４－メトキシベンジル、又はＴＢＤＰＳ、ＴＢＳ、ＴＥＳ、若しくはＴＩＰＳなどのアルキル若しくはアリールシランから選択される）と、触媒ＣｕＩ及びジアミン、例えば、トランス－１，２－ジアミノシクロヘキサン、及び塩基、例えば、Ｋ_３ＰＯ_４、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの存在下、１，４－ジオキサン、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で、Ｕｌｌｍａｎｎ型芳香族アミノ化反応させて、式（ＶＩ）の化合物（式中、ＸがＣＨであり、ＹがＣＨである）を得る。

According to Scheme 3, a compound of formula (V), where R ¹ is an optionally substituted C _2-6 alkenyl, R ³ is H, R ⁴ is a suitably substituted phenyl as described herein with reference to formula (Z), and HAL is Br, can be reacted with a commercially available or synthetically available nucleophilic compound of formula (II), where R ^a is a C _{1-6 alkyl, for example a suitably protected triazolone, where PG is selected from benzyl, 4-methoxybenzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES, or TIPS, in the presence of the catalyst CuI and a diamine, for example trans-1,2} -diaminocyclohexane, and a base, for example K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , in the presence of triethylamine or the like in a suitable solvent such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc., to give a compound of formula (VI) (wherein X is CH and Y is CH).

A compound of formula (VI) where PG is Bn and R ¹ is C _2-6 alkenyl is reacted under Simmons-Smith cyclopropanation reaction conditions known to one skilled in the art to obtain a compound of formula (VI) where R ¹ is C _3-6 cycloalkyl substituted with C _1-6 alkyl. For example, a compound of formula (VI) where R ¹ is

である）を、ジヨードメタン、及びジエチル亜鉛と、トルエンなどの好適な溶媒中、０℃～室温の範囲の温度で、３～２６時間かけて反応させて、式（ＶＩ）の化合物（Ｒ^１がＣＨ_３で置換されたシクロプロピルである）を得る。

(R 1 is cyclopropyl substituted with CH 3) is reacted with diiodomethane and diethylzinc in a suitable solvent such as toluene at a temperature ranging from 0° C. to room temperature for 3 to 26 hours to give the compound of formula (VI) (R ¹ is cyclopropyl substituted with CH ₃ ).

Deprotection using established methods such as those described in T. W. Greene and P. G. M. Wuts, "Protective Groups in Organic Synthesis", 3rd Edition, John Wiley & Sons, 1999, then gives the compound of formula (Z) where X and Y are CH. For example, the compound of formula (VI) where R ³ is H and PG is TBDPS is deprotected using conditions known to those skilled in the art, preferably with TBAF in a suitable solvent such as THF. In a preferred method, PG is TBDPS and R ^a is C _1-6 alkyl. Alternatively, removal of the TBDPS protecting group is accomplished using triethylamine trihydrogen fluoride (Et ₃ N.3HF).

Removal of the Bn protecting group is accomplished in the presence of hydrogen gas in the presence of a catalyst such as palladium on carbon (Pd/C). Removal of the Bn protecting group is also accomplished using TFA at a temperature of about 80°C.

The compound of formula (Z), where X is CH, Y is CH, R ² , R ³ , R ⁴ are each defined as described herein for formula (Z); R ¹ is C _2-6 alkenyl, is reduced using hydrogenation conditions known to those skilled in the art, for example reaction with Pd/C or Wilkinson's catalyst [RhCl(PPh ₃ ) ₃ ] under H ₂ in a suitable solvent such as MeOH, THF, EtOAc, etc., to obtain the compound of formula (Z), where R ¹ is C _2-6 alkyl.

Scheme 4

スキーム４に従って、式（ＶＩＩ）の化合物を、α、β－不飽和アルデヒド、例えば、３－メチル－２－ブテナール、３－メチルペント－２－エナールなどと、－ＴｉＣｌ_４、及びトリエチルアミンなどの塩基を使用して、ジクロロメタン（ＤＣＭ）などの非プロトン性溶媒中で還元的アミノ化して、エナミン中間体を得た後、ＮａＢＨ_４などの還元剤を用いて還元して、式（ＶＩＩＩ）（式中、Ｒ^５はＣ_１～４アルキルであり、Ｒ^４は式（Ｚ）に関して本明細書で定義される通りである）の化合物を得る。式（ＶＩＩＩ）の化合物を、トリエチルアミン及び４－ジメチルアミノピリジン（ＤＭＡＰ）などの塩基を用いて市販の又は合成により入手可能な４－ブロモ－２－ヨードベンゾイルクロリドと、ジクロロメタン（ＤＣＭ）などの無水非プロトン性溶媒中でカップリングさせて、式（ＩＸ）の化合物を得る。加熱ヘック反応条件下で式（ＩＸ）の化合物を酢酸パラジウム（ＩＩ）、臭化テトラブチルアンモニウム及び酢酸カリウムで処理して、式（Ｖ）の分子内環化化合物（式中、Ｒ^１は任意選択的に置換されているＣ_２～６アルキルであり、Ｒ^３はＨであり、ＸはＣＨであり、ＨＡＬはＢｒである）、及び、式（Ｖａ）の分子内環化化合物（式中、Ｒ^１は任意選択的に置換されているＣ_２～６アルケニルであり、Ｒ^３はＨであり、ＸはＣＨ_２であり、ＨＡＬはＢｒである）を得る。

According to Scheme 4, the compound of formula (VII) is reductively aminated with an α,β-unsaturated aldehyde, for example, 3-methyl-2-butenal, 3-methylpent-2-enal, etc., using -TiCl ₄ and a base such as triethylamine in an aprotic solvent such as dichloromethane (DCM) to give an enamine intermediate, which is then reduced with a reducing agent such as NaBH ₄ to give a compound of formula (VIII), where R ⁵ is C _1-4 alkyl and R ⁴ is as defined herein for formula (Z). The compound of formula (VIII) is coupled with commercially available or synthetically available 4-bromo-2-iodobenzoyl chloride using triethylamine and a base such as 4-dimethylaminopyridine (DMAP) in an anhydrous aprotic solvent such as dichloromethane (DCM) to give a compound of formula (IX). Treatment of the compound of formula (IX) with palladium(II) acetate, tetrabutylammonium bromide and potassium acetate under heated Heck reaction conditions gives the intramolecularly cyclized compound of formula (V) where R ¹ is an optionally substituted C _2-6 alkyl, R ³ is H, X is CH and HAL is Br, and the intramolecularly cyclized compound of formula (Va) where R ¹ is an optionally substituted C _2-6 alkenyl, R ³ is H, X is CH ₂ and HAL is Br.

Scheme 5

スキーム５に従って、市販の又は合成により入手可能な式（Ｘ）の化合物（式中、ＨＡＬがＦであり、Ｒ^３がＦであり、かつＲ^５がＨ又はＣ_１～４アルキルである）を、市販の又は合成により入手可能な式（ＩＩ）の求核性化合物（式中、Ｒ^ａはＣ_１～６アルキル、例えば、適切に保護されたトリアゾロンであり、ＰＧは、ベンジル、４－メトキシベンジル、又はＴＢＤＰＳ、ＴＢＳ、ＴＥＳ、若しくはＴＩＰＳなどのアルキル若しくはアリールシランから選択される）と、Ｋ_３ＰＯ_４、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの塩基の存在下、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で反応させて、式（ＸＩ）の化合物を得る。好ましい方法では、ＰＧはＢｎであり、Ｒ^ａはＣ_１～６アルキルである。式（ＸＩ）のエステルは、Ｒ^５がＣ_１～４アルキルである場合、酸性又は塩基性条件下で、その対応する酸に加水分解される。例えば、ｔｅｒｔ－ブチルエステル（Ｒ^５はｔｅｒｔ－Ｂｕである）をＴＦＡで処理するか、あるいは、水性溶媒中、ＮａＯＨのような塩基で加水分解すると、式（ＸＩａ）の化合物（式中、Ｒ^５がＨである）が得られる。式（ＸＩａ）の化合物を、当業者に公知の条件を用いて塩素化して、式（ＸＩＩ）の塩化アシルを得る。例えば、式（ＸＩａ）の化合物をＳＯＣｌ_２中で加熱するか、又はＤＣＭ中の塩化オキサリルで処理する。

According to scheme 5, a commercially available or synthetically available compound of formula (X), where HAL is F, R ³ is F and R ⁵ is H or C _1-4 alkyl, is reacted with a commercially available or synthetically available nucleophilic compound of formula (II), where R ^a is C _1-6 alkyl, e.g. a suitably protected triazolone, and PG is selected from benzyl, 4-methoxybenzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES or TIPS, in the presence of a base such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, in a suitable solvent such as DMSO, DMF, THF, ACN, to give a compound of formula (XI). In a preferred method, PG is Bn and R ^a is C _1-6 alkyl. Esters of formula (XI), when R ⁵ is C _1-4 alkyl, may be hydrolyzed to their corresponding acids under acidic or basic conditions. For example, tert-butyl esters (R ⁵ is tert-Bu) may be treated with TFA or hydrolyzed with a base such as NaOH in an aqueous solvent to give compounds of formula (XIa) where R ⁵ is H. Compounds of formula (XIa) may be chlorinated using conditions known to those skilled in the art to give acyl chlorides of formula (XII). For example, compounds of formula (XIa) may be heated in SOCl ₂ or treated with oxalyl chloride in DCM.

Scheme 6

スキーム６に従って、式（ＸＩＩ）の化合物（式中、Ｒ^３がＨ又はＦであり、ＰＧがＢｎであり、Ｒ^ａがＣ_１～６アルキルである）を、式（ＶＩＩＩ）の化合物（式中、Ｒ^５がＣ_１～４アルキルである）と、トリエチルアミン（ＴＥＡ）及び４－ジメチルアミノピリジン（ＤＭＡＰ）の混合物などの塩基を用いて、ジクロロメタン（ＤＣＭ）などの無水非プロトン性溶媒中で反応させて、式（ＸＩＩＩ）の化合物を得る。式（ＶＩ）の化合物（式中、ＸがＣＨであり、ＹがＣＨである）は、式（ＸＩＩＩ）の化合物（式中、Ｒ^１が、任意選択的に、式（Ｚ）に関して本明細書に記載されているように置換されるＣ_１～６アルキルである）を、酢酸パラジウム（ＩＩ）、臭化テトラブチルアンモニウム、及び酢酸カリウムと、加熱ヘック反応条件下で反応させて、分子内環化化合物の混合物を得、次いでこれを分離して、Ｒ^１がＣ_２～６アルキルであり、Ｒ^３がＨ又はＦである中間体化合物を単離する。

According to Scheme 6, a compound of formula (XII), wherein R ³ is H or F, PG is Bn, and R ^a is C _1-6 alkyl, is reacted with a compound of formula (VIII), wherein R ⁵ is C _1-4 alkyl, using a base such as a mixture of triethylamine (TEA) and 4-dimethylaminopyridine (DMAP) in an anhydrous aprotic solvent such as dichloromethane (DCM) to obtain a compound of formula (XIII). A compound of formula (VI), where X is CH and Y is CH, can be prepared by reacting a compound of formula (XIII), where R ¹ is C _1-6 alkyl, optionally substituted as described herein for formula (Z), with palladium(II) acetate, tetrabutylammonium bromide, and potassium acetate under heated Heck reaction conditions to give a mixture of intramolecularly cyclized compounds, which are then separated to isolate intermediate compounds, where R ¹ is C _2-6 alkyl and R ³ is H or F.

Scheme 7

スキーム７に従って、式（ＸＩＶ）の化合物（Ｒ^３がＨ又はＦである）を、式（ＩＩ）の化合物、例えば、好適に保護されたトリアゾロン（式中、ＰＧは、ベンジル、４－メトキシベンジル、又はＴＢＤＰＳ、ＴＢＳ、ＴＥＳ、若しくはＴＩＰＳなどのアルキル若しくはアリールシランから選択される）と、前述の方法に従って、Ｕｌｌｍａｎｎ型芳香族アミノ化反応させて、式（ＸＶ）の化合物を得る。好ましい方法では、ＰＧはＢｎであり、Ｒ^ａはＣ_１～６アルキルである。式（ＸＶ）の化合物を、Ｎ－ヨードスクシンイミド（ＮＩＳ）などのハロゲン化試薬で、アセトニトリルなどの非プロトン性溶媒中、加熱条件下で処理して、式（ＸＶＩ）のハロゲン化化合物（式中、ＹがＣＨであり、ＨＡＬがヨウ化物である）を得る。

According to Scheme 7, a compound of formula (XIV) (R ³ is H or F) is reacted with a compound of formula (II), for example a suitably protected triazolone, where PG is selected from benzyl, 4-methoxybenzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES, or TIPS, by Ullmann-type aromatic amination according to the methods previously described to give a compound of formula (XV). In a preferred method, PG is Bn and R ^a is a C _1-6 alkyl. The compound of formula (XV) is treated with a halogenating reagent, such as N-iodosuccinimide (NIS), in an aprotic solvent such as acetonitrile under heating conditions to give a halogenated compound of formula (XVI), where Y is CH and HAL is an iodide.

Scheme 8

スキーム８に従って、式（ＸＶＩＩａ）及び（ＸＶＩＩｂ）の化合物が、５－ブロモイソベンゾフラン－１，３－ジオンから２工程で調製される。５－ブロモイソベンゾフラン－１，３－ジオンを、ｉ－ＰｒＭｇＣｌ、ＥｔＭｇＢｒなどの市販の又は合成により入手可能な適切に置換されたアルキルグリニャール試薬と、ＣｄＣｌ_２の存在下、ＴＨＦなどの非プロトン性溶媒中で反応させ、続いて、Ｒ^５がＣ_１～４アルキルである式Ｒ^５－Ｉのアルキル化剤（ヨードメタン又はヨードエタンなど）で、例えば、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３などの好適に選択された塩基の存在下、ＤＭＦ、ＤＭＳＯなどの非プロトン性溶媒中で処理して、式（ＸＶＩＩａ）及び（ＸＶＩＩｂ）の位置異性体エステル（式中、Ｒ^１が任意選択的に置換されているＣ_１～６アルキルである）の混合物を得る。同様に、アリールグリニャール試薬を使用して、式（ＸＶＩＩａ）及び（ＶＸＩＩｂ）の化合物（式中、Ｒ^１が適切に置換されたフェニルである）を得ることができる。式（ＸＶＩＩａ）及び（ＸＶＩＩｂ）の位置異性体は分離されていないが、直接使用され、対応するフタラジノン（混合物）に変換される。例えば、式（ＸＶＩＩａ）及び式（ＸＶＩＩｂ）の混合物を過剰のヒドラジンで、エタノール又はメタノールなどの適切な溶媒中、室温～９０℃の範囲の温度で、６～２０時間かけて処理する。式（Ｖ）の所望のフタラジノン化合物は、沈殿、結晶化、又はフラッシュクロマトグラフィにより精製することによって、他の位置異性体から容易に分離することができる。式（Ｖ）の化合物と、式（ＩＩ）（式中、Ｒ^ａはＣ_１～６アルキルであり、ＰＧは、ベンジル、４－メトキシベンジル、又はＴＢＤＰＳ、ＴＢＳ、ＴＥＳ、若しくはＴＩＰＳなどのアルキル若しくはアリールシランから選択される）の適切に保護されたトリアゾロンとを、触媒ＣｕＩ及びジアミン、例えば、トランス－１，２－ジアミノシクロヘキサン、及び塩基、例えば、Ｋ_３ＰＯ_４、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの存在下、１，４－ジオキサン、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で、Ｕｌｌｍａｎｎ型芳香族アミノ化反応させて、式（ＸＶＩＩＩ）の化合物（式中、ＸはＮである）を得る。

According to Scheme 8, compounds of formula (XVIIa) and (XVIIb) are prepared in two steps from 5-bromoisobenzofuran-1,3-dione. Reaction of 5-bromoisobenzofuran-1,3-dione with a commercially available or synthetically available appropriately substituted alkyl Grignard reagent such as i-PrMgCl, EtMgBr in the presence of CdCl ₂ in an aprotic solvent such as THF, followed by treatment with an alkylating agent of formula R ⁵ -I, where R ⁵ is C _1-4 alkyl, such as iodomethane or iodoethane, in the presence of a suitably selected base, such as K ₂ CO ₃ , Cs ₂ CO ₃ , in an aprotic solvent such as DMF, DMSO, gives a mixture of regioisomeric esters of formula (XVIIa) and (XVIIb), where R ¹ is optionally substituted C _1-6 alkyl. Similarly, aryl Grignard reagents can be used to obtain compounds of formula (XVIIa) and (VXIIb), where R ¹ is an appropriately substituted phenyl. The regioisomers of formula (XVIIa) and (XVIIb) are not separated but are used directly and converted to the corresponding phthalazinones (mixture). For example, a mixture of formula (XVIIa) and formula (XVIIb) is treated with excess hydrazine in a suitable solvent such as ethanol or methanol at temperatures ranging from room temperature to 90° C. for 6 to 20 hours. The desired phthalazinone compound of formula (V) can be easily separated from the other regioisomers by precipitation, crystallization, or purification by flash chromatography. Ullmann type aromatic amination reaction of compound of formula (V) with a suitably protected triazolone of formula (II) (wherein R ^a is C _1-6 alkyl and PG is selected from benzyl, 4-methoxybenzyl, or alkyl or aryl silanes such as TBDPS, TBS, TES, or TIPS) in the presence of catalyst CuI and a diamine such as trans-1,2-diaminocyclohexane and a base such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc. in a suitable solvent such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc., gives compound of formula (XVIII) (wherein X is N).

The compound of formula (XVIII), where R ¹ is a C _2-6 alkenyl, is reacted under Simmons-Smith cyclopropanation reaction conditions known to one skilled in the art to provide a compound of formula (XVIII), where R ^{1 is a C 3-6 cycloalkyl substituted with a C 1-6 alkyl. For example, the compound of formula (XVIII), where R 1} _{is a C 2-6} ^alkenyl _, is reacted with diiodomethane and diethylzinc in a suitable solvent, such as toluene, at a temperature ranging from 0° C. to room temperature for 24 to 26 hours to provide a compound of formula (XVIII), where R ¹ is a cyclopropyl substituted with CH ₃ .

Scheme 9

スキーム９に従って、式（Ｘ）の化合物（式中、ＨＡＬはＢｒであり、Ｒ^３はＨであり、Ｒ^５はＣＨ_３である）を、パラジウム触媒カルボニル化反応において、式Ｒ^１－ＣＨＯ（式中、Ｒ^１はＣ_１～６アルキルである）の市販の又は合成により入手可能なアルデヒドとカップリングさせて、式（ＸＩＸ）の対応するケトン化合物を得る（同様の変換がＳｕｃｈａｎｄら、Ｊ．Ｏｒｇ．Ｃｈｅｍ．２０１６，８１，６４０９－６４２３に報告されている）。例えば、メチル４－ブロモ－２－ヨード安息香酸とイソブチルアルデヒドを、Ｐｄ（ＯＡｃ）_２などのパラジウム触媒、Ａｇ_２Ｏ、及びｔｅｒｔ－ブチルヒドロペルオキシド（ＴＢＨＰ）の水溶液などの酸化剤の存在下、約１２０℃の温度で、１０～１４時間かけて反応させて、メチル４－ブロモ－２－イソブチリルベンゾエートを得た。式（ＸＩＸ）のケトン化合物をヒドラジンＲ^４－ＮＨＮＨ_２（式中、Ｒ^４は適切に置換されたアリール、例えば、２－クロロ－６－フルオロフェニルヒドラジンである）と反応させ、式（Ｖ）（式中、ＸはＮである）の化合物を得る。式（Ｖ）の化合物と、前述の適切に保護されたトリアゾロン（ＩＩ）とのＵｌｌｍａｎｎ型芳香族アミノ化反応により、式（ＶＩ）（式中、ＹはＣＨであり、Ｒ^１はＣ_１～６アルキルから選択される）の化合物を得る。

According to Scheme 9, a compound of formula (X), where HAL is Br, R ³ is H, and R ⁵ is CH ₃ , is coupled with a commercially available or synthetically available aldehyde of formula R ¹ -CHO, where R ¹ is C _1-6 alkyl, in a palladium catalyzed carbonylation reaction to give the corresponding ketone compound of formula (XIX). (A similar transformation has been reported in Suchand et al., J. Org. Chem. 2016, 81, 6409-6423.) For example, methyl 4-bromo-2-iodobenzoate and isobutyraldehyde are reacted in the presence of a palladium catalyst such as Pd(OAc) ₂ , Ag ₂ O, and an oxidizing agent such as an aqueous solution of tert-butyl hydroperoxide (TBHP) at a temperature of about 120° C. for 10-14 hours to give methyl 4-bromo-2-isobutyrylbenzoate. Reaction of the ketone compound of formula (XIX) with hydrazine R ⁴ -NHNH ₂ , where R ⁴ is a suitably substituted aryl, for example, 2-chloro-6-fluorophenylhydrazine, gives the compound of formula (V), where X is N. Ullmann-type aromatic amination reaction of the compound of formula (V) with the suitably protected triazolone (II) as described above gives the compound of formula (VI), where Y is CH and R ¹ is selected from C _1-6 alkyl.

Compounds of formula (VI) where Y is CH, R ¹ is phenyl and X is N can be prepared in a similar manner using the methods described above by coupling methyl 4-bromo-2-iodobenzoate with commercially available or synthetically available aldehydes of formula R ¹ -CHO where R ¹ is phenyl.

Scheme 10

スキーム１０に従って、式Ｒ^１－Ｂ（ＯＨ）_２の化合物を、当業者に周知の鈴木カップリング条件下で、式（ＸＶＩ）の化合物と反応させて、式（ＸＶＩＩＩ）の化合物（式中、ＸはＣＨである）を得る。例えば、式（ＸＶＩ）の化合物（式中、ＹがＣＨであり、ＨＡＬがヨウ化物である）を、市販の又は合成により入手可能なボロン酸（又はボロン酸エステル）、例えば、Ｒ^１－Ｂ（ＯＨ）_２（式中、Ｒ^１は、式（Ｚ）を参照して本明細書で定義されている任意選択的に置換されているＣ_２～６アルケニル、Ｃ_３～６シクロアルキル又はアリールである）、ビス（トリフェニルホスフィン）パラジウム（ＩＩ）ジクロリドなどのパラジウム触媒、リン酸カリウム、Ｃｓ_２ＣＯ_３などの適切な塩基と、ジオキサン、水、エタノール、又はこれらの混合物などの好適な溶媒中で反応させて、式化合物（ＸＶＩＩＩ）の化合物（式中、ＸがＣＨである）を得る。上記のカップリング反応中、反応条件中のヨウ化物の損失により、式化合物（ＸＶＩＩＩ）の化合物（式中、ＸはＣＨであり、Ｒ^１はＨである）が得られたことがわかった。式化合物（ＸＶＩＩＩ）の化合物（式中、ＸはＣＨ又はＮである）は、式Ｒ^４－Ｂ（ＯＨ）_２の化合物と、当業者に公知の銅（ＩＩ）媒介Ｃｈａｎ－Ｌａｍカップリング条件下で、又は前述のように反応させて、式（ＶＩ）の化合物（式中、ＸはＣＨ又はＮであり、Ｒ^１は任意選択的に置換されているＣ_２～６アルケニルであり、Ｒ^３はＨ又はＦであり、Ｒ^４は式（Ｚ）を参照して本明細書に記載されるような適切に置換されたフェニルである）を得る。

According to Scheme 10, a compound of formula R ¹ -B(OH) ₂ is reacted with a compound of formula (XVI) under Suzuki coupling conditions well known to those skilled in the art to give a compound of formula (XVIII) where X is CH. For example, a compound of formula (XVI) where Y is CH and HAL is iodide is reacted with a commercially available or synthetically available boronic acid (or boronic ester), for example R ¹ -B(OH) ₂ where R ¹ is optionally substituted C _2-6 alkenyl, C _3-6 cycloalkyl or aryl as defined herein with reference to formula (Z), a palladium catalyst such as bis(triphenylphosphine)palladium(II) dichloride, potassium phosphate, a suitable base such as Cs ₂ CO ₃ , in a suitable solvent such as dioxane, water, ethanol, or a mixture thereof to give a compound of formula (XVIII) where X is CH. It was found that during the above coupling reaction, loss of iodide during the reaction conditions afforded a compound of formula compound (XVIII) where X is CH and R ¹ is H. A compound of formula compound (XVIII) where X is CH or N can be reacted with a compound of formula R ⁴ -B(OH) ₂ under copper(II) mediated Chan-Lam coupling conditions known to those skilled in the art or as previously described to afford a compound of formula (VI) where X is CH or N, R ¹ is an optionally substituted C _2-6 alkenyl, R ³ is H or F and R ⁴ is an appropriately substituted phenyl as described herein with reference to formula (Z).

Compounds of formula (XVIII) where R ¹ is N(CH ₃ ) ₂ are prepared from compounds of formula (XVI) where HAL is Br and PG is Bn. Compounds of formula (XVI) are reacted with amine bs, for example NH(CH ₃ ) ₂ , in water at a temperature of about 110° C. for 96 hours to obtain compounds of formula (XVIII) where R ¹ is N(CH ₃ ) ₂ and R ^a is C _1-6 alkyl. Compounds of formula (Z) where R ¹ is N(CH ₃ ) ₂ are prepared according to the above method.

The compound of formula (XVIII) where R ¹ is C _1-6 alkyl substituted with OH is prepared in two steps from the compound of formula (XVIII) where R ¹ is C _2-6 alkenyl and PG is Bn. In the first step, the compound of formula (XVIII) where R ¹ is

である）を、例えば、ＮａＩＯ_４、及びＫ_２ＯｓＯ_４．２Ｈ_２Ｏ又はＯｓＯ_４などの酸化条件下、ＴＨＦ／Ｈ_２Ｏなどの好適な溶媒中、０℃～室温の範囲の温度で、４８～７２時間かけて反応させて、ケトン中間体化合物を得る。第２の工程では、ケトン中間体化合物を、臭化メチルマグネシウムなどのグリニャール試薬と、ジエチルエーテルなどの適切な溶媒中、０℃～室温の範囲の温度で、３～３０時間かけて反応させて、式（ＸＶＩＩＩ）の化合物（式中、Ｒ^１がＯＨで置換されたＣ_１～６アルキルである）を得る。

(XVIII) is reacted with a Grignard _reagent such as methylmagnesium bromide in a suitable solvent such as diethyl ether _at a temperature ranging from 0° C. to room temperature for 3-30 hours under oxidizing conditions such as NaIO ₄ and K ₂ OsO _{4.2H 2} O or OsO 4 to give a ketone intermediate compound. In a second step, the ketone intermediate compound is reacted with a Grignard reagent such as methylmagnesium bromide in a suitable solvent such as diethyl ether at a temperature ranging from 0° C. to room temperature for 3-30 hours to give a compound of formula (XVIII) where R ¹ is a C _1-6 alkyl substituted with OH.

Scheme 11

スキーム１１に従って、４，５－ジフルオロフタル酸無水物を、式Ｒ^４－ＮＨＮＨ_２のヒドラジン化合物（式中、Ｒ^４は、適切に置換されたフェニル又はヘテロアリール、例えば、（２－クロロ－６－フルオロフェニル）ヒドラジン塩酸塩である）と、酢酸中、約１２５℃の温度で、約１．５時間かけて反応させて、式（ＸＸ）（式中、Ｒ^３はＦである）の化合物を得る。式（ＸＸ）の化合物を、ナトリウムエトキシド又はナトリウムメトキシドなどの塩基性条件下、エタノール、メタノールなどの好適な溶媒中、室温で、約１．５時間の期間にわたって再構成することにより、式（ＸＸＩ）の環拡張化合物を得る。トリフルオロメタンスルホニル（トリフレート）などのスルホネート系脱離基を有する式（ＸＸＩ）の化合物の誘導は、ＤＣＭなどの適切な溶媒中、トリフルオロメタンスルホン酸無水物（Ｔｆ_２Ｏ）などのトリフレート剤、トリエチルアミン（ＴＥＡ）などの塩基、ピリジンなどと反応させて、式（ＸＸＩＩ）の化合物を得ることによって達成される。Ｎ－フェニルビス（トリフルオロメタンスルホンイミド）（ＴＦ_２ＮＰｈ）などのより穏やかなトリフレート化剤と、ＴＥＡ、ＤＩＥＡなどの塩基を、ＤＣＭなどの好適な溶媒中で、使用することができる。

According to Scheme 11, 4,5-difluorophthalic anhydride is reacted with a hydrazine compound of formula R ⁴ -NHNH ₂ , where R ⁴ is an appropriately substituted phenyl or heteroaryl, for example, (2-chloro-6-fluorophenyl)hydrazine hydrochloride, in acetic acid at a temperature of about 125° C. for about 1.5 hours to give a compound of formula (XX), where R ³ is F. Reconstitution of the compound of formula (XX) under basic conditions such as sodium ethoxide or sodium methoxide in a suitable solvent such as ethanol, methanol at room temperature for a period of about 1.5 hours gives a ring-expanded compound of formula (XXI). Derivatization of compounds of formula (XXI) with a sulfonate-based leaving group such as trifluoromethanesulfonyl (triflate) is accomplished by reaction with a triflating agent such as trifluoromethanesulfonic anhydride (Tf ₂ O), a base such as triethylamine (TEA), pyridine, etc. in a suitable solvent such as DCM to give compounds of formula (XXII). Milder triflating agents such as N-phenylbis(trifluoromethanesulfonimide) (TF ₂ NPh) and bases such as TEA, DIEA, etc. in a suitable solvent such as DCM can be used.

Scheme 12

スキーム１２に従って、式Ｒ^１－Ｂ（ＯＨ）_２の化合物を、前述の鈴木カップリングの条件下、式（ＸＸＩＩ）の化合物と反応させて、式（Ｖ）の化合物（式中、ＸはＮである）を得る。例えば、式（ＸＸＩＩ）の化合物を、市販の又は合成により入手可能なボロン酸（又はボロン酸エステル）、例えば、Ｒ^１－Ｂ（ＯＨ）_２（式中、Ｒ^１は式（Ｚ）を参照して本明細書で定義されるＣ_２～６アルケニル又はＣ_２～６ハロアルケニルである）、ビス（トリフェニルホスフィン）パラジウム（ＩＩ）ジクロリド、又は１，１’－ビス（ジフェニルホスフィノ）フェロセン－パラジウム（ＩＩ）ジクロリドジクロロメタン錯体などのパラジウム触媒、及びリン酸カリウム、Ｃｓ_２ＣＯ_３Ｋ_２ＣＯ_３などの適切な塩基と、ジオキサン、水、エタノール、又はこれらの混合物などの好適な溶媒中で反応させて、式化合物（Ｖ）の化合物を得る。式（Ｖ）の化合物（式中、Ｒ^１がＣ_２～６アルキル又はＣ_２～６ハロアルキルである）は、式（Ｖ）の化合物（式中、Ｒ^１がＣ_２～６アルケニル又はＣ_２～６ハロアルケニルである）の選択的水素化によって容易に調製される。例えば、式（Ｖ）の化合物（式中、Ｒ^１が

According to Scheme 12, a compound of formula R ¹ -B(OH) ₂ is reacted with a compound of formula (XXII) under Suzuki coupling conditions as described above to give a compound of formula (V), where X is N. For example, a compound of formula (XXII) is reacted with a commercially available or synthetically available boronic acid (or boronic ester), for example R ¹ -B(OH) ₂ (where R ¹ is C _2-6 alkenyl or C _2-6 haloalkenyl as defined herein with reference to formula (Z)), a palladium catalyst such as bis(triphenylphosphine)palladium(II) dichloride or 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex, and a suitable base such as potassium phosphate, Cs ₂ CO ₃ K ₂ CO ₃ in a suitable solvent such as dioxane, water, ethanol, or a mixture thereof to give a compound of formula (V). Compounds of formula (V) where R ¹ is C _2-6 alkyl or C _2-6 haloalkyl are readily prepared by selective hydrogenation of compounds of formula (V) where R ¹ is C _2-6 alkenyl or C _2-6 haloalkenyl. For example, compounds of formula (V) where R ¹ is

である）を、Ｐｄ／Ｃなどの触媒を使用する水素化条件下、ＥｔＯＡｃなどの適切な溶媒中で、水素ガス（２０～４５ｐｓｉ）雰囲気下、室温で、４～２４時間かけて反応させて、式（Ｖ）の化合物（式中、Ｒ^１が

is reacted under hydrogenation conditions using a catalyst such as Pd/C in a suitable solvent such as ^EtOAc under an atmosphere of hydrogen gas (20-45 psi) at room temperature for 4-24 hours to give a compound of formula (V),

である）を得る。前述の条件を用いた、式（Ｖ）の化合物と、式（ＩＩ）の適切に保護されたトリアゾロンとの反応により、式（ＶＩ）及び（ＶＩａ）の化合物の混合物が得られ、これは、保護基の脱保護の前又は後に分離することができる。

Reaction of a compound of formula (V) with a suitably protected triazolone of formula (II) using the conditions described above gives a mixture of compounds of formula (VI) and (VIa), which can be separated before or after deprotection of the protecting groups.

Scheme 13

スキーム１３に従って、式（ＸＶＩＩＩ）の化合物のＮ－アリール化は、適切に置換された市販の又は合成により入手可能な式（ＸＸＩＩＩ）のフルオロ化合物（式中、式中、Ｒ^ｃ及びＲ^ｄは、式（Ｚ）に関して本明細書において定義されている通りである）の反応によって達成される。式（ＸＶＩＩＩ）の化合物（式中、Ｒ^１がＨ、Ｃ_２～６アルケニル、Ｃ_２～６ハロアルケニル、Ｃ_３～６シクロアルキル、Ｃ_１～６アルキルで置換されたＣ_３～６シクロアルキルであり、ＸがＣＨ又はＮである）を、求核置換反応条件下、市販の又は合成により入手可能な式（ＸＸＩＩＩ）のフルオロ化合物と、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３などの塩基の存在下、ＤＭＦ、ＤＭＳＯなどの非プロトン性溶媒中、６５～１００℃の範囲の温度で反応させて、式（ＸＸＩＶ）の化合物を得る。

According to Scheme 13, N-arylation of compounds of formula (XVIII) is achieved by reaction of appropriately substituted commercially or synthetically available fluoro compounds of formula (XXIII), where R ^c and R ^d are as defined herein for formula (Z). Compounds of formula (XVIII), where R ¹ is H, C _2-6 alkenyl, C _2-6 haloalkenyl, C _3-6 cycloalkyl, C 3-6 cycloalkyl substituted with C _1-6 alkyl, and X is CH or N, are reacted under nucleophilic substitution reaction conditions with commercially or synthetically available fluoro compounds of formula (XXIII) in the presence of a base such as K ₂ CO ₃ , Cs ₂ CO ₃ , in an aprotic solvent such as _DMF , DMSO, at temperatures ranging from 65 to 100° C. to give compounds of formula (XXIV).

Reduction of compounds of formula (XXIV) is accomplished using zinc or iron and NH ₄ Cl in a mixed solvent of methanol and water to give amino compounds of formula (XXV).

Diazotization of compound of formula (XXV) with _NaNO2 in aqueous acid or other nitrite reagent in organic solvent such as EtOH at a temperature of 0° C., followed by reduction of the diazo group with zinc at a temperature ranging from 0 to 85° C. or treatment with _H3PO2 gives compound of formula ( _XXVI ), where ^Rc and ^Rd are as defined herein for formula (Z).

Scheme 14

スキーム１４に従って、式（Ｘ）の化合物（式中、ＨＡＬがＦであり、Ｒ^５がＨであり、Ｒ^３がＦである）を、市販の又は合成により入手可能な式（ＸＸＶＩＩ）の化合物（式中、Ｒ^１ａ及びＲ^１ｂがそれぞれ独立してＨ又はＣ_１～４アルキルである）、例えば、１－ブロモ－３－メチル－２－ブテンと、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３などの塩基の存在下、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で反応させて、式（ＸＸＶＩＩＩ）のエステル化合物（式中、Ｒ^３がＦであり、ＨＡＬがＦである）を得る。式（ＸＸＶＩＩＩ）の化合物（式中、Ｒ^１ａ及びＲ^１ｂは、Ｃ_１～４ハロアルキル又はＣ_３～６シクロアルキルからそれぞれ独立して選択される）は、同様の方法で作製することができる。式（ＸＸＶＩＩＩ）のエステルを、適切に保護された式（ＩＩ）のトリアゾロン化合物と、Ｋ_３ＰＯ_４、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの塩基の存在下、１，４－ジオキサン、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で、Ｕｌｌｍａｎｎ型芳香族アミノ化反応させて、式（ＸＸＩＸ）の化合物を得る。好ましい方法では、ＰＧはＢｎであり、Ｒ^ａはＣ_１～６アルキルである。式（ＸＸＩＸ）の化合物（式中、Ｒ^３がＨ又はＦである）は、トルエンなどの適切な溶媒中、クロロ［（トリ－ｔｅｒｔ－ブチルホスフィン）－２－（２－アミノビフェニル）］パラジウム（ＩＩ）（Ｐ（ｔＢｕ_３）ＰｄＧ_２）、Ｎ－シクロヘキシル－Ｎ－メチル－シクロヘキサンアミンなどの触媒を使用するといった、ヘック反応条件下で、約１５～８０℃の温度にて、約１８～３６時間かけて、分子内環化を行って、式（ＸＸＸ）のイソクマリン化合物（式中、ＹがＣＨであり、Ｒ^１がイソプロピルであり、Ｒ^３がＨ又はＦであり、Ｒ^ａ及びＰＧが上記に定義される通りである）を得る。

According to Scheme 14, a compound of formula (X), where HAL is F, R ⁵ is H and R ³ is F, is reacted with a commercially available or synthetically available compound of formula (XXVII), where R ^1a and R ^1b are each independently H or C _1-4 alkyl, for example 1-bromo-3-methyl-2-butene, in the presence of a base such as K ₂ CO ₃ , Cs ₂ CO ₃ , in a suitable solvent such as DMSO, DMF, THF, ACN, to give an ester compound of formula (XXVIII), where R ³ is F and HAL is F. Compounds of formula (XXVIII), where R ^1a and R ^1b are each independently selected from C _1-4 haloalkyl or C _3-6 cycloalkyl, can be prepared in a similar manner. The ester of formula (XXVIII) undergoes Ullmann type aromatic amination reaction with a suitably protected triazolone compound of formula (II) in the presence of a base such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc., in a suitable solvent such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc., to give the compound of formula (XXIX). In a preferred process, PG is Bn and R ^a is C _1-6 alkyl. The compound of formula (XXIX), wherein R ³ is H or F, undergoes intramolecular cyclization under Heck reaction conditions, such as using a catalyst such as chloro[(tri-tert-butylphosphine)-2-(2-aminobiphenyl)]palladium(II) (P(tBu ₃ )PdG ₂ ), N-cyclohexyl-N-methyl-cyclohexanamine, in a suitable solvent such as toluene, at a temperature of about 15-80° C. for about 18-36 hours to provide the isocoumarin compound of formula (XXX), wherein Y is CH, R ¹ is isopropyl, R ³ is H or F, and R ^a and PG are as defined above.

Isocoumarin compounds of formula (XXX ⁾

である）は、式（ＸＩａ）の化合物及びメチルブタ－１，２－ジエン－１－イルアセテートから調製される。メチルブタ－１，２－ジエン－１－イルアセテートは、市販されており、又は２－メチル－３－ブチン－２－オールから２工程で調製される。無水酢酸を、２－メチル－３－ブチン－２－オールと、Ｍｇ（ＣｌＯ_４）_２などの触媒の存在下、ＤＣＭなどの好適な溶媒中、反応させて、２－メチルブタ－３－イン－２－イルアセテートを得る。２－メチルブタ－３－イン－２－イルアセテートを触媒量のＡｇＢＦ_４、ＡｇＣｌＯ_４、ＰｔＣｌ_２などのルイス酸と反応させて、３－メチルブタ－１，２－ジエン－１－イルアセテートを得る。３－メチルブタ－１，２－ジエン－１－イルアセテートは、前述のようなヘック反応条件下で分子間環化を用い、式（ＸＩａ）の化合物（式中、Ｒ^５はＨである）とカップリングさせる。例えば、Ｃａｔａｃｘｉｕｍ Ａ Ｐｄ Ｇ２、及びＣｙ_２ＮＭｅパラジウム（ＩＩ）などの触媒、テトラブチルアンモニウムブロミドのような相転移試薬、及び酢酸カリウムのような塩基を用い、ＤＭＦなどの好適な溶媒中、７０～９０℃の温度で、１０～１６時間かけて反応させて、式（ＸＸＸ）のイソクマリン化合物（式中、ＹはＣＨであり、Ｒ^１は

is prepared from a compound of formula (XIa) and methylbut-1,2-dien-1-yl acetate, which is commercially available or prepared in two steps from 2-methyl-3-butyn-2-ol. Acetic anhydride is reacted with 2-methyl-3-butyn-2-ol in the presence of a catalyst such as Mg(ClO ₄ ) ₂ in a suitable solvent such as DCM to give 2-methylbut-3-yn-2-yl acetate. Reaction of 2-methylbut-3-yn-2-yl acetate with a catalytic amount of a Lewis acid such as AgBF ₄ , AgClO ₄ , PtCl ₂ gives 3-methylbut-1,2-dien-1-yl acetate. 3-Methylbuta-1,2-dien-1-yl acetate is coupled with a compound of formula (XIa) (wherein R ⁵ is H) using intermolecular cyclization under Heck reaction conditions as previously described. For example, the isocoumarin compound of formula (XXX) (wherein Y is CH and R 1 is H) can be obtained by reaction using a catalyst such as Catacxium A Pd G2 and Cy ₂ NMe palladium (II), a phase transfer reagent such as tetrabutylammonium bromide, and a base such as potassium acetate in a suitable solvent such as DMF at a temperature of 70-90° C. for ^10-16 hours.

である）を得る。

) is obtained.

Compounds of formula (XXX) where Y is CH and R ¹ is

である）は、ウィルキンソン触媒［ＲｈＣｌ（ＰＰｈ_３）_３］などの触媒を用いる水素化条件下、ＴＨＦなどの適切な溶媒中、室温で、選択的に還元して、式（ＸＸＸ）のイソクマリン化合物（式中、Ｒ^１がイソプロピルである）を得る。

(XXX) is selectively reduced under hydrogenation conditions using a catalyst such as Wilkinson's catalyst [RhCl( _PPh3 ) ₃ ] in a suitable solvent such as THF at room temperature to give isocoumarin compounds of formula (XXX), where ^R1 is isopropyl.

Scheme 15

スキーム１５に従って、当業者に周知のウィッティヒ反応条件を用いて、２－ブタノンをエチル３－メチルペンタ－２－エノエートに変換する。例えば、２－ブタノンを、（カルベトキシメチレン）トリフェニルホスホランなどのトリフェニルホスホニウムイリドと、安息香酸、ＬｉＣｌ及びドデシル硫酸ナトリウム（ＳＤＳ）などの添加剤を用いて又は用いずに、トルエンなどの適切な溶媒中、室温から溶媒の還流温度までの範囲の温度で１２～２４時間かけて反応させる。エチル３－メチルペント－２－エノエートは、トルエンなどの適切な溶媒中、－７８℃～室温の範囲の温度で、ＤＩＢＡＬ－Ｈなどの適切な還元剤を用いて、３－メチルペント－２－エン－１－オールに還元される。３－メチルペンタ－２－エン－１－オールは、当業者に周知の酸化条件、例えば、ＥｔＯＡｃ、ＤＭＳＯ、ＤＣＭなどの溶媒中、約－７８℃～室温（約２３℃）の範囲の温度で、ＤＭＰ（デス・マーチン・ペルヨージナン）、ＳＯ_３－ピリジン、スワーン条件［（ＣＯＣｌ）_２、ＤＭＳＯ、Ｅｔ_３Ｎ］、ＰＣＣなどを使用して、３－メチルペンタ－２－エナールに酸化させる。好ましい方法では、３－メチルペンタ－２－エン－１－オールを、デス・マーチン・ペルヨージナンと共に、ＤＣＭ中で、２５℃で１～４時間酸化させ、３－メチルペンタ－２－エナールにする。

According to Scheme 15, 2-butanone is converted to ethyl 3-methylpent-2-enoate using Wittig reaction conditions well known to those skilled in the art. For example, 2-butanone is reacted with a triphenylphosphonium ylide, such as (carbethoxymethylene)triphenylphosphorane, with or without additives, such as benzoic acid, LiCl, and sodium dodecyl sulfate (SDS), in a suitable solvent, such as toluene, at a temperature ranging from room temperature to the reflux temperature of the solvent for 12-24 hours. Ethyl 3-methylpent-2-enoate is reduced to 3-methylpent-2-en-1-ol using a suitable reducing agent, such as DIBAL-H, in a suitable solvent, such as toluene, at a temperature ranging from −78° C. to room temperature. 3-Methylpent-2-en-1-ol is oxidized to 3-methylpent-2-enal using oxidation conditions well known to those skilled in the art, for example, using DMP (Dess-Martin periodinane), SO ₃ -pyridine, Swern conditions [(COCl) ₂ , DMSO, Et ₃ N], PCC, etc., in solvents such as EtOAc, DMSO, DCM, and at temperatures ranging from about −78° C. to room temperature (about 23° C.). In a preferred method, 3-methylpent-2-en-1-ol is oxidized with Dess-Martin periodinane in DCM at 25° C. for 1-4 hours to 3-methylpent-2-enal.

Scheme 16

スキーム１６に従って、式（ＸＸＸ）の化合物（式中、ＹがＣＨである）のイソクマリンを、市販の又は合成により入手可能な式Ｒ^４－ＮＨ_２のアミン化合物（式中、Ｒ^４は、式（Ｚ）に関して本明細書で定義した通りである）、及びＡｌＭｅ_３、ＡｌＣｌ_３などのルイス酸と、ＤＣＭ、トルエンなどの適切な非プロトン性溶媒中、反応させて、式（ＸＸＸＩ）の化合物（式中、ＹがＣＨであり、Ｒ^１、Ｒ^３、Ｒ^４及びＲ^ａが式（Ｚ）を参照して本明細書に記載されるように定義される）を得る。

According to Scheme 16, isocoumarin compounds of formula (XXX), where Y is CH, are reacted with commercially available or synthetically available amine compounds of formula R ^-NH , where ^R is as defined herein for formula (Z ₎ , and Lewis acids such as _AlMe , _AlCl , in a suitable aprotic solvent such as DCM, toluene, to give compounds of formula (XXXI), where Y is CH and ^R , ^R , ^R and ^R are defined as described herein with reference to formula (Z).

Scheme 17

式（ＸＸＸ）のイソクマリン化合物は、スキーム１７により調製することができる。４，５－ジフルオロ－２－ヨード安息香酸クロリドは、塩化オキサリル又は塩化チオニルなどの当業者に周知の条件を用いて、触媒量のＤＭＦの存在下で、ジクロロメタン（ＤＣＭ）、テトラヒドロフラン（ＴＨＦ）、アセトニトリル（ＡＣＮ）、トルエンなどの非プロトン性非極性溶媒などの好適な溶媒中で、０℃～室温の範囲の温度で、式（Ｘ）の化合物（式中、ＨＡＬはＦであり、Ｒ^５はＨであり、Ｒ^３はＦである）の化合物から調製され、４，５－ジフルオロ－２－ヨード安息香酸クロリドを形成する。４，５－ジフルオロ－２－ヨード安息香酸クロリドを、トリエチルアミン及びＤＭＡＰなどの塩基の存在下、市販の又は合成により入手可能な２－メチルブト－３－イン－２－オールと、ＤＣＭなどの好適な溶媒中、反応させて、式（ＸＸＸＩＩＩ）のエステル化合物を得ることができる。前述した方法を使用して、式（ＸＸＸＩＩＩ）の化合物を式（ＩＩ）の化合物と反応させることにより、式（ＸＸＸＩＶ）の化合物を得ることができる。式（ＸＸＸＩＶ）の化合物を触媒量のルイス酸、例えば、ＡｇＣｌＯ_４、ＰｔＣｌ_２などで処理して、式（ＸＸＸＶ）の転位された化合物を得ることができる。式（ＸＸＸＶ）の化合物（式中、Ｒ^３はＨ又はＦである）は、酢酸パラジウム（ＩＩ）などの触媒、テトラブチルアンモニウムブロミドのような相転移試薬、及び酢酸カリウムのような塩基を使用するなど、ＤＭＦなどの好適な溶媒中、ヘック反応条件下で、７０～９０℃の温度で、１～３時間かけて、分子内環化させ得、式（ＸＸＸ）のイソクマリン化合物（式中、ＹはＣＨである）を得る。

Isocoumarin compounds of formula (XXX) can be prepared according to Scheme 17. 4,5-Difluoro-2-iodobenzoic acid chloride can be prepared from compounds of formula (X) (wherein HAL is F, R 5 is H, and R 3 is F) using conditions well known to those skilled in the art such as oxalyl chloride or thionyl chloride in the presence of catalytic ^amount of DMF in a suitable solvent such as aprotic non-polar solvents such as dichloromethane (DCM), ^{tetrahydrofuran} (THF), acetonitrile (ACN), toluene, etc., at temperatures ranging from 0° C. to room temperature to form 4,5-difluoro-2-iodobenzoic acid chloride. 4,5-Difluoro-2-iodobenzoic acid chloride can be reacted with commercially available or synthetically available 2-methylbut-3-yn-2-ol in the presence of a base such as triethylamine and DMAP in a suitable solvent such as DCM to give ester compounds of formula (XXXIII). Using the methods previously described, compounds of formula (XXXIII) can be reacted with compounds of formula (II) to give compounds of formula (XXXIV). Compounds of formula (XXXIV) can be treated with catalytic amounts of Lewis acids, such as AgClO ₄ , PtCl ₂ and the like, to give rearranged compounds of formula (XXXV). Compounds of formula (XXXV) (wherein R ³ is H or F) can be intramolecularly cyclized under Heck reaction conditions, such as using a catalyst such as palladium (II) acetate, a phase transfer reagent such as tetrabutylammonium bromide, and a base such as potassium acetate, in a suitable solvent such as DMF, at a temperature of 70-90° C. for 1-3 hours to give isocoumarin compounds of formula (XXX) (wherein Y is CH).

Scheme 18

スキーム１８に従って、式（Ｘａ）の化合物（式中、ＨＡＬがＣｌであり、Ｒ^５がＣＨ（ＣＨ_３）_２であり、Ｒ^３がＦである）は、市販されているか、又はＣｈｅｎら、米国特許出願公開第２０１６－０１７６８６９号に記載の方法に従って合成により入手可能である。式（Ｘａ）の化合物を、市販の又は合成により入手可能な式（ＩＩ）の求核性化合物（式中、ＰＧはベンジルであり、Ｒ^ｃはＣ_１～６アルキルである）と、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの好適な塩基の存在下、ジメチルスルホキシド（ＤＭＳＯ）、ＤＭＦ、ＴＨＦ、ＡＣＮなどの好適な溶媒中で反応させて、式（ＸＸＸＩＸ）（式中、ＹはＮである）の化合物を得る。式（ＸＸＸＩＸ）又は式（ＸＩ）の化合物（式中、Ｒ^３はＦであり、Ｒ^５はＣ_１～４アルキルである）を、市販の１－エトキシエテン－２－ボロン酸ピナコールエステル、ビス（トリフェニルホスフィン）パラジウム（ＩＩ）ジクロリド、１，１’－ビス（ジフェニルホスフィノ）フェロセン－パラジウム（ＩＩ）ジクロリドなどのパラジウム触媒、及びＣｓ_２ＣＯ_３などの適切な塩基と、ジオキサン、水、エタノール、又はこれらの混合物などの好適な溶媒中、従来の加熱法又はマイクロ波加熱法を用いて反応させて、式（ＸＬ）の化合物（式中、Ｙは、Ｎ又はＣＨである）を得る。

According to Scheme 18, compounds of formula (Xa) where HAL is Cl, ^R5 is CH( _CH3 ) ₂ and ^R3 is F are either commercially available or synthetically accessible according to the methods described in Chen et al., US Patent Publication No. 2016-0176869. Compounds of formula (Xa) are reacted with commercially available or synthetically accessible nucleophilic compounds _{of formula (II) where PG is benzyl and Rc is C1-6 alkyl in the presence of a suitable base such as K2CO3, Cs2CO3} ^, _{NaHCO3 , triethylamine ,} in a suitable solvent such as dimethylsulfoxide (DMSO), DMF, THF, ACN _, to provide compounds of formula (XXXIX) where Y is N. The compound of formula (XXXIX) or formula (XI), where R ³ is F and R ⁵ is C _1-4 alkyl, is reacted with commercially available 1-ethoxyethene-2-boronic acid pinacol ester, a palladium catalyst such as bis(triphenylphosphine)palladium(II) dichloride, 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, and a suitable base such as Cs ₂ CO ₃ in a suitable solvent such as dioxane, water, ethanol, or a mixture thereof, using conventional or microwave heating methods to provide the compound of formula (XL), where Y is N or CH.

Scheme 19

スキーム１９によれば、式Ｒ^４－ＮＨ_２の化合物（式中、Ｒ^４は式（Ｚ）を参照して本明細書で定義される通りである）を、トリメチルアルミニウムと、ジクロロメタン、トルエン、又はこれらの混合物などの好適な溶媒中で反応させ、得られた溶液を、式（ＸＬ）の化合物（式中、ＹはＣＨ又はＮである）と合わせて、式（ＸＬＩ）の化合物を得る。式（ＸＬＩ）の化合物（式中、ＹはＣＨ又はＮである）を、５０℃～９０℃の加熱条件下で酢酸又はトリフルオロ酢酸で処理して、式（ＸＬＩＩ）の化合物を得る。式（ＸＬＩＩ）の化合物を、Ｎ－ブロモスクシンイミドを使用して、無水ジメチルホルムアミド中、室温でハロゲン化して、式（ＸＶＩ）の化合物（式中、ＨＡＬがＢｒである）を得る。式Ｒ^１－Ｂ（ＯＨ）_２の化合物を、当業者に公知の鈴木カップリング条件下で、又は前述したように、式（ＸＶＩ）の化合物と反応させて、式（ＶＩ）の化合物（式中、Ｒ^１は、任意選択的に置換されているＣ_２～６アルケニル、Ｃ_２～６ハロアルケニル、又は式（Ｚ）に関して本明細書で定義されるアリールである）を得る。式（ＶＩ）の化合物（式中、Ｒ^１が任意選択的に置換されているＣ_２～６アルケニル又はＣ_２～６ハロアルケニルである）を、水素化条件下、ウィルキンソン触媒（（ＰＰｈ_３）_３ＲｈＣｌ）を用いて反応させて、式（ＶＩ）の化合物（式中、Ｒ^１がＣ_２～６アルキル又はＣ_２～６ハロアルキルである）を得る。

According to Scheme 19, a compound of formula R ⁴ -NH ₂ , where R ⁴ is as defined herein with reference to formula (Z), is reacted with trimethylaluminum in a suitable solvent such as dichloromethane, toluene, or a mixture thereof, and the resulting solution is combined with a compound of formula (XL), where Y is CH or N, to obtain a compound of formula (XLI). The compound of formula (XLI), where Y is CH or N, is treated with acetic acid or trifluoroacetic acid under heating conditions at 50°C to 90°C to obtain a compound of formula (XLII). The compound of formula (XLII) is halogenated using N-bromosuccinimide in anhydrous dimethylformamide at room temperature to obtain a compound of formula (XVI), where HAL is Br. The compound of formula R ¹ -B(OH) ₂ is reacted with a compound of formula (XVI) under Suzuki coupling conditions known to one skilled in the art or as previously described to provide a compound of formula (VI) where R ¹ is an optionally substituted C _2-6 alkenyl, C _2-6 haloalkenyl, or aryl as defined herein for formula (Z). The compound of formula (VI) where R ¹ is an optionally substituted C 2-6 alkenyl or C _2-6 haloalkenyl is reacted with Wilkinson's catalyst ((PPh ₃ ) ₃ RhCl) under hydrogenation conditions to provide a compound of formula (VI) where R ¹ is _{a C 2-6 alkyl or C 2-6 haloalkyl .}

Scheme 20

スキーム２０に従って、３－メチルブタナールを、アリルパラジウム（ＩＩ）クロリド二量体などのパラジウム触媒を用いて、式（ＸＸＸＩＸ）の化合物（式中、ＹがＮであり、Ｒ^５がＣＨ（ＣＨ_３）_２である）、１，１’－ビス（ジフェニルホスフィノ）フェロセン（ｄｐｐｆ）などの配位子、及びＣｓ_２ＣＯ_３などの適切な塩基と、モレキュラーシーブ（４Ａ）などの水捕捉剤の存在下、ジオキサンなどの適切な溶媒中で反応させて、式化合物（ＸＸＸ）の化合物（式中、Ｒ^１がイソプロピルである）を得る。式Ｒ^４－ＮＨ_２の化合物（式中、Ｒ^４は、式（Ｚ）に関して本明細書において定義されている通りである）を、トリメチルアルミニウムと、ジクロロメタン、トルエン、又はこれらの混合物などの好適な溶媒中で反応させ、得られた溶液を、式（ＸＸＸ）の化合物と合わせて、続いて、８０～１００℃の加熱温度で、５～２４時間の範囲の時間をかけて酢酸で処理することにより、式（ＶＩ）の化合物（式中、ＸはＣＨであり、ＹはＮであり、Ｒ^１はイソプロピルであり、Ｒ^３はＦである）を得る。

According to Scheme 20, 3-methylbutanal is reacted with a compound of formula (XXXIX) where Y is N and R ⁵ is CH(CH ₃ ) ₂ , a ligand such as 1,1′-bis(diphenylphosphino)ferrocene (dppf), and a suitable base such as Cs ₂ CO ₃ using a palladium catalyst such as allylpalladium(II) chloride dimer in the presence of a water scavenger such as molecular sieves (4A) in a suitable solvent such as dioxane to give a compound of formula (XXX) where R ¹ is isopropyl. A compound of formula R ⁴ -NH ₂ , where R ⁴ is as defined herein for formula (Z), is reacted with trimethylaluminum in a suitable solvent such as dichloromethane, toluene, or mixtures thereof, and the resulting solution is combined with a compound of formula (XXX), followed by treatment with acetic acid at a heating temperature of 80-100°C for a period ranging from 5 to 24 hours to obtain a compound of formula (VI), where X is CH, Y is N, R ¹ is isopropyl, and R ³ is F.

Scheme 21

スキーム２１に従って、式（ＸＸＸＩＸ）又は式（ＸＩ）の化合物（式中、Ｒ^３がＦであり、Ｒ^５がＣ_１～４アルキルである）を、市販のビニルボロン酸ピナコールエステル、ビス（トリフェニルホスフィン）パラジウム（ＩＩ）ジクロリド、又は１，１’－ビス（ジフェニルホスフィノ）フェロセン－パラジウム（ＩＩ）ジクロリドジクロロメタン錯体などのパラジウム触媒、及びＣｓ_２ＣＯ_３などの適切な塩基と、ジオキサン、水、エタノール、又はこれらの混合物などの好適な溶媒中で反応させて、式化合物（（ＸＬＩＩＩ）（式中、Ｙは、Ｎ又はＣＨである）の化合物を得る。式（ＸＬＩＩＩ）の化合物中のビニル基は、オスミウム酸カリウム（ＶＩ）二水和物／過ヨウ素酸ナトリウム、又はオゾン分解などを用いて、式（ＸＬＩＶ）のアルデヒド基に選択的に変換される。式（ＸＬＩＶ）の化合物を、市販の又は合成により入手可能な、好適に置換されているアルキルグリニャール試薬（ｉ－ＰｒＭｇＣｌなど）と、ＴＨＦなどの非プロトン性溶媒中で反応させ、続いて、デス・マーチン試薬、又はスワーン酸化条件などの酸化試薬による後続の処理を行うことにより、式（ＸＬＶ）のケトン化合物を得る。

According to Scheme 21, a compound of formula (XXXIX) or formula (XI) where R ³ is F and R ⁵ is C _1-4 alkyl can be reacted with commercially available vinylboronic acid pinacol ester, a palladium catalyst such as bis(triphenylphosphine)palladium(II) dichloride, or 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex, and Cs ₂ CO ₃ in a suitable solvent such as dioxane, water, ethanol, or a mixture thereof to give a compound of formula (XLIII) where Y is N or CH. The vinyl group in the compound of formula (XLIII) is selectively converted to an aldehyde group of formula (XLIV) using, for example, potassium osmate (VI) dihydrate/sodium periodate, or ozonolysis. Reaction of the compound of formula (XLIV) with a suitably substituted alkyl Grignard reagent, either commercially available or available by synthesis, such as i-PrMgCl, in an aprotic solvent such as THF, followed by subsequent treatment with an oxidation reagent such as the Dess-Martin reagent or Swern oxidation conditions, gives the ketone compound of formula (XLV).

Compounds of formula (XLV) are prepared in two steps from compounds of formula (XXXIX). Compounds of formula (XXXIX), where ^R3 is F and ^R5 is _C1-4 alkyl, are reacted with commercially available tributyl(1-ethoxyvinyl)tin and a palladium catalyst, such as bis(triphenylphosphine)palladium(II) dichloride, 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, in a suitable solvent, such as dioxane, water, ethanol, or mixtures thereof. Subsequent acidic hydrolysis, using conditions such as treatment with aqueous HCl at room temperature, provides compounds of formula (XLV), where X is N, Y is N or CH, and ^R1 is methyl.

Commercially or synthetically available hydrazine R ⁴ -NHNH ₂ , where R ⁴ is as defined herein with reference to formula (Z), for example 2-chloro-6-fluorophenylhydrazine, o-tolylhydrazine, is condensed with a compound of formula (XLV) in the presence of a base such as potassium carbonate under heating conditions such as at 70-120° C. in a suitable solvent such as toluene or mixtures thereof to give a compound of formula (VI), where X is N, Y is CH or N and R ⁴ is as defined herein with reference to formula (Z).

Scheme 22

スキーム２２に従って、２－ブロモ－４，５－ジフルオロ安息香酸メチルを、適切に保護された式（ＩＩ）のトリアゾロン化合物と、Ｋ_３ＰＯ_４、Ｋ_２ＣＯ_３、Ｃｓ_２ＣＯ_３、ＮａＨＣＯ_３、トリエチルアミンなどの塩基の存在下、１，４－ジオキサン、ＤＭＳＯ、ＤＭＦ、ＴＨＦ、ＡＣＮなどの適切な溶媒中で反応させて、式（ＸＬＶＩ）の化合物を得る。好ましい方法では、ＰＧはＢｎであり、Ｒ^ａはＣ_１～６アルキルである（スキーム１４で前述した通り）。３－メチルブタナールを式（ＸＬＶＩ）の化合物を、アリルパラジウム（ＩＩ）クロリド二量体などのパラジウム触媒を用いて、１，１’－ビス（ジフェニルホスフィノ）フェロセン（ｄｐｐｆ）などの配位子、及びＣｓ_２ＣＯ_３などの適切な塩基と、モレキュラーシーブ（４Ａ）などの水捕捉剤の非存在下、ジオキサンなどの適切な溶媒中で反応させて、式化合物（ＸＬＶＩＩ）の化合物を得る。式Ｒ^４－ＮＨ_２の化合物（式中、Ｒ^４は、式（Ｚ）に関して本明細書において定義されている通りである）を、トリメチルアルミニウムと、ジクロロメタン、ジクロロエタン、トルエン、又はこれらの混合物などの好適な溶媒中で反応させ、得られた溶液を、式（ＸＬＶＩＩ）の化合物と合わせて、続いて、８０～１００℃の加熱温度で、５～２４時間の範囲の時間をかけて酢酸で処理することにより、式（ＶＩ）の化合物（式中、ＸはＣＨであり、ＹはＣＨであり、Ｒ^１はイソプロピルであり、Ｒ^３はＦである）を得る。場合によって、ｏ－トルイジンなどの式Ｒ^４－ＮＨ_２の化合物を、８０～１００℃の加熱温度で、１０～２４時間の範囲の時間にわたって、酢酸中で式化合物（ＸＬＶＩＩ）の化合物と直接縮合させて、式（ＶＩ）の化合物（式中、Ｘ、Ｙ、Ｒ^１、Ｒ^３は先に定義されている）を得る。

According to Scheme 22, methyl 2-bromo-4,5-difluorobenzoate is reacted with a suitably protected triazolone compound of formula (II) in the presence of a base such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc. in a suitable solvent such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc. to give the compound of formula (XLVI). In a preferred method, PG is Bn and R ^a is C _1-6 alkyl (as previously described in Scheme 14). Reaction of 3-methylbutanal with a compound of formula (XLVI) using a palladium catalyst such as allylpalladium(II) chloride dimer, a ligand such as 1,1′-bis(diphenylphosphino)ferrocene (dppf), and a suitable base such as Cs ₂ CO ₃ in a suitable solvent such as dioxane in the absence of a water scavenger such as molecular sieves (4A) provides a compound of formula (XLVII). A compound of formula R ⁴ -NH ₂ , where R ⁴ is as defined herein for formula (Z), is reacted with trimethylaluminum in a suitable solvent such as dichloromethane, dichloroethane, toluene, or mixtures thereof, and the resulting solution is combined with a compound of formula (XLVII) and subsequently treated with acetic acid at a heating temperature of 80-100° C. for a time ranging from 5 to 24 hours to obtain a compound of formula (VI), where X is CH, Y is CH, R ¹ is isopropyl, and R ³ is F. In some cases, a compound of formula R ⁴ -NH ₂ , such as o-toluidine, is directly condensed with a compound of formula (XLVII) in acetic acid at a heating temperature of 80-100° C. for a time ranging from 10 to 24 hours to obtain a compound of formula (VI), where X, Y, R ¹ , R ³ are defined above.

Scheme 23

スキーム２３に従って、式（ＸＸＸＩＸ）の化合物（式中、Ｒ^５がＣ_１～４アルキルであり、ＹがＮであり、Ｒ^３がＦである）を、トリメチルシリルアセチレンなどのシリル保護アルキン、パラジウム（ＩＩ）ビス（トリフェニルホスフィン）ジクロリドなどのパラジウム触媒、ヨウ化銅などの銅触媒を用い、トリエチルアミンなどの適切な塩基を用いて、ＡＣＮ、トルエンなどの適切な溶媒中で、薗頭カップリング反応に供する。ＴＢＡＦを用い、ＴＨＦなどの適切な溶媒中、室温で脱保護反応を行って、式（ＸＬＶＩＩＩ）の化合物を得る。式（ＸＬＩＸ）の化合物は、ＭｅＣＮなどの適切な溶媒混合物中、金触媒、好ましくはＡｕＣｌ_３を使用して得られる。ＡｕＣｌ_３触媒環化による同様の変換が、Ｍａｒｃｈａｌ，Ｅ．らによってＴｅｔｒａｈｅｄｒｏｎ ２００７，６３，９９７９－９９９０に記載されている。式Ｒ^４－ＮＨ_２の化合物（式中、Ｒ^４は、式（Ｚ）に関して本明細書において定義されている通りである）を、トリメチルアルミニウムと、ジクロロメタン、ジクロロエタン、トルエン、又はこれらの混合物などの好適な溶媒中で反応させ、得られた溶液を、式（ＸＬＩＸ）の化合物と合わせて、続いて、８０～１００℃の加熱温度で、５～２４時間の範囲の時間をかけて酢酸で処理することにより、式（Ｌ）の化合物を得る。ＮＢＳを、室温で、好適な溶媒（好ましくはＤＭＦ）中で使用した後、当業者に周知の条件を用いたクロスカップリングを行い、好ましくはパラジウム（ＩＩ）ビス（トリフェニルホスフィン）ジクロリドなどのパラジウム触媒、Ｃｓ_２ＣＯ_３、Ｎａ_２ＣＯ_３などの塩基を用い、１，４－ジオキサン及び水からなる溶媒混合物中で、１００℃の温度で反応させて、式（ＶＩ）の化合物（式中、ＸがＣＨであり、ＹがＮであり、Ｒ^３がＦであり、Ｒ^１、Ｒ^ａ及びＰＧが前述のように定義される）を得る。

According to scheme 23, compounds of formula (XXXIX) where ^R5 is _C1-4 alkyl, Y is N and ^R3 is F are subjected to Sonogashira coupling reaction using a silyl protected alkyne such as trimethylsilylacetylene, a palladium catalyst such as palladium(II) bis(triphenylphosphine) dichloride, a copper catalyst such as copper iodide, and a suitable base such as triethylamine in a suitable solvent such as ACN, toluene, etc. Deprotection reaction is carried out using TBAF in a suitable solvent such as THF at room temperature to give compounds of formula (XLVIII). Compounds of formula (XLIX) are obtained using a gold catalyst, preferably _AuCl3 , in a suitable solvent mixture such as MeCN. A similar transformation by _AuCl3 catalyzed cyclization is described by Marchal, E. et al. in Tetrahedron 2007, 63, 9979-9990. A compound of formula R ⁴ -NH ₂ , where R ⁴ is as defined herein for formula (Z), is reacted with trimethylaluminum in a suitable solvent such as dichloromethane, dichloroethane, toluene, or mixtures thereof, and the resulting solution is combined with a compound of formula (XLIX) followed by treatment with acetic acid at a heating temperature of 80-100°C for a time period ranging from 5 to 24 hours to obtain a compound of formula (L). NBS is used at room temperature in a suitable solvent, preferably DMF, followed by cross-coupling using conditions well known to those skilled in the art, preferably using a palladium catalyst such as palladium(II) bis(triphenylphosphine) dichloride, a base such as Cs ₂ CO ₃ , Na ₂ CO ₃ , in a solvent mixture consisting of 1,4-dioxane and water at a temperature of 100° C. to give a compound of formula (VI), where X is CH, Y is N, R ³ is F, and R ¹ , R ^a and PG are as defined above.

Scheme 24

スキーム２４に従って、式（ＶＩ）の化合物（式中、ＰＧはＢｎである）を、当業者に周知の条件を用いて、好ましくは密封管内で未希釈のＴＦＡ中で、約６０～９０℃の温度で、又は、ＢＣｌ_３を用いて、約－７８℃の温度で、ＤＣＭ中などの適切な溶媒中で脱保護を行い、又は、パラジウム炭素（Ｐｄ／Ｃ）などの触媒の存在下で、水素ガスで処理することにより、式（Ｚ）の化合物を得る。

According to Scheme 24, compounds of formula (VI), where PG is Bn, are deprotected using conditions well known to one skilled in the art, preferably in a sealed tube in neat TFA at a temperature of about 60-90° C., or with BCl ₃ at a temperature of about −78° C. in a suitable solvent such as DCM, or by treatment with hydrogen gas in the presence of a catalyst such as palladium on carbon (Pd/C), to give compounds of formula (Z).

In a similar manner, N-arylation and in situ TBDPS deprotection of compounds of formula (XVIII) where ^R1 is I, PG is TBDPS, and X is N is accomplished using conditions well known to those skilled in the art or previously described to provide compounds of formula (Z).

A compound of formula (Z) where ^R3 is F is reacted in an aromatic nucleophilic substitution reaction to give a compound of formula (Z) where ^R3 is _OCH3 . For example, a compound of formula (Z) where ^R3 is F is reacted with a suitable base such as NaOH in a suitable solvent such as MeOH to give a compound of formula (Z) where Y is CH and ^R3 is _OCH3 .

Compounds of formula (Z) may be converted to their corresponding salts using methods known to those skilled in the art. For example, amines of formula (Z) are treated with trifluoroacetic acid, HCl, or citric acid in solvents such as Et ₂ O, CH ₂ Cl ₂ , THF, MeOH, chloroform, or isopropanol to obtain the corresponding salt forms. Alternatively, reverse-phase HPLC purification conditions result in trifluoroacetate or formate salts. Crystalline forms of pharma-ceutically acceptable salts of compounds of formula (Z) can be obtained in crystalline form by recrystallization from polar solvents (including mixtures of polar solvents and aqueous mixtures of polar solvents) or non-polar solvents (including mixtures of non-polar solvents).

When the compounds according to the invention have at least one chiral center, they may consequently exist as enantiomers. When the compounds have two or more chiral centers, they may additionally exist as diastereomers. It is understood that all such isomers and mixtures thereof are encompassed within the scope of the present invention.

Compounds prepared according to the above schemes can be obtained in a single form, such as a single enantiomer, by form-specific synthesis or by resolution. Alternatively, compounds prepared according to the above schemes can be obtained as a mixture of various forms, such as racemic (1:1) or non-racemic (non-1:1) mixtures. When racemic and non-racemic mixtures of enantiomers are obtained, single enantiomers can be isolated using conventional isolation methods known to those skilled in the art, such as chiral chromatography, recrystallization, diastereomeric salt formation, derivatization to diastereomeric adducts, biotransformation, or enzymatic conversion. When regioisomeric or diastereomeric mixtures are obtained, single isomers can be separated using conventional methods, such as chromatography or crystallization, as appropriate.

The following specific examples further illustrate the compounds of formula (Z) and various preferred embodiments.

In obtaining the compounds of formula (Z) and the corresponding analytical data described in the examples below, the following experimental and analytical protocols were followed unless otherwise indicated.

Unless otherwise noted, reaction mixtures were magnetically stirred at room temperature (rt) under _a nitrogen atmosphere. When solutions were "dried," they were generally dried over a drying agent such as _Na2SO4 or _MgSO4 . When mixtures, solutions, and extracts were "concentrated," they were typically concentrated on a rotary evaporator under reduced pressure.

Normal phase silica gel chromatography (FCC) was performed on silica gel (SiO ₂ ) using prepacked cartridges.

Preparative reversed-phase high performance liquid chromatography (RP HPLC) was performed in one of the following ways.
Method A. Gilson GX-281 semi-preparative HPLC with a Phenomenex Synergi C18 (10 μm, 150×25 mm) or Boston Green ODS C18 (5 μm, 150×30 mm) and a mobile phase of 5-99% ACN in water (with 0.225% FA) at a flow rate of 25 mL/min over 10 min, followed by a 2 min hold at 100% ACN. Or Method B. Gilson GX-281 semi-preparative HPLC (Phenomenex Synergi C18 (10 μm, 150×25 mm), or Boston Green ODS C18 (5 μm, 150×30 mm) and a mobile phase of 5-99% ACN in water (0.1% TFA) held for 10 min, then 100% ACN for 2 min at a flow rate of 25 mL/min. or Method C. Gilson GX-281 semi-preparative HPLC (Phenomenex Synergi C18 (10 μm, 150×25 mm), or Boston Green ODS C18 (5 μm, 150×30 mm) and a mobile phase of 5-99% ACN in water (0.05% HCl) held for 10 min, then 100% ACN for 2 min at a flow rate of 25 mL/min. ACN mobile phase held for 10 min, then 100% ACN for 2 min at a flow rate of 25 mL/min. OR Method D. Gilson GX-281 semi-preparative HPLC with a Phenomenex Gemini C18 (10 μm, 150×25 mm), AD (10 μm, 250 mm×30 mm), or Waters XBridge C18 column (5 μm, 150×30 mm), mobile phase of 0-99% ACN in water (with 0.05% ammonium hydroxide v/v) at a flow rate of 25 mL/min for 10 min, then 100% ACN for 2 min. OR Method E. Phenomenex Gemini C18 (10 μm, 150×25 mm) or Waters XBridge C18 column (5 μm, 150×30 mm), mobile phase of 0-99% ACN in water (with 0.05% ammonium hydroxide v/v), at a flow rate of 25 mL/min for 10 min, then 100% ACN for 2 min. Gilson GX-281 semi-preparative HPLC equipped with a C18 column (5 μm, 150×30 mm), flow rate 25 mL/min, mobile phase of 5-99% ACN in water (10 mM NH ₄ HCO ₃ ) over 10 min, then hold at 100% ACN for 2 min.

Preparative supercritical fluid high performance liquid chromatography (SFC) was performed on either a Thar 80 Prep-SFC system or a Waters 80Q Prep-SFC system from Waters. The ABPR was set at 100 bar to maintain CO2 at SF conditions, and flow rates ranged from 50 g/min to 70 g/min, which could be verified according to compound properties. Column temperature was ambient.

Unless otherwise stated, mass spectra (MS) were obtained on a SHIMADZU LCMS-2020 MSD or an Agilent 1200\G6110A MSD using electrospray ionization (ESI) in positive mode. Calculated masses (calcd.) correspond to exact masses.

Nuclear magnetic resonance (NMR) spectra were obtained on a Bruker AVIII 400 spectrometer. Multiplicity definitions are as follows: s = singlet, d = doublet, t = triplet, q = quartet, dd = double doublet, ddd = double doublet doublet, td = triple doublet, dt = double triplet, spt = septet, quin = quintet, m = multiplet, br = broad line. It will be understood that in compounds that contain exchangeable protons, the protons may or may not be visible in the NMR spectrum, depending on the choice of solvent used to perform the NMR spectrum and the concentration of the compounds in solution.

Chemical names were generated using ChemDraw Ultra 12.0, ChemDraw Ultra 14.0 (CambridgeSoft Corp., Cambridge, MA) or ACD/Name Version 10.01 (Advanced Chemistry). Compounds designated R ^* or S ^* are enantiomerically pure compounds for which the absolute configuration has not been determined.

Intermediate 1: 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one.

Step A. 2-(Benzyloxy)acetohydrazide.
To a solution of ethyl 2-(benzyloxy)acetate (55 g, 283.17 mmol) in EtOH (500 mL) was added NH ₂ NH _{2.H 2} O (28.3 g, 566 mmol, 27.5 mL). The reaction mixture was heated at 78° C. for 6 h. The reaction mixture was concentrated under reduced pressure to give the title product (52 g, crude) as a colorless oil, which was used directly in the next step without further purification.

Step B. 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one.
To a solution of 2-(benzyloxy)acetohydrazide (52 g, 288 mmol) in H ₂ O (500 mL) was added isocyanatoethane (25.1 g, 346 mmol, 27.9 mL) dropwise at 0° C. After the addition was complete, the mixture was stirred at 25° C. for 12 h. To the mixture was added H ₂ O (20 mL) and a solution of NaOH (57.7 g, 1.44 mol) in water (120 mL). The mixture was stirred at 95° C. for 12 h. The reaction mixture was cooled to room temperature and then quenched with HCl (12 M) at 0° C. and the pH was adjusted to 6. The solid was filtered and dried under reduced pressure to give the title compound as a white solid (61 g, 91% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ -9.23-9.09 (m, 1H), -7.41-7.31 (m, 5H), -4.58-4.53 (m, 2H), -4.45-4.42 (m, 2H), -3.82-3.75 (m, 2H), -1.33-1.29 (m, 3H) ppm.

Intermediate 2: 5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one.

Step A. 4-Ethyl-5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a solution of 5-[(benzyloxy)methyl]-4-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (8 g, 34.3 mmol, 1.0 equiv.) in methanol (200 mL) was added Pd/C (2 g). The resulting mixture was kept under hydrogen and stirred at room temperature for 6 h. The resulting mixture was then filtered and the filtrate was concentrated to give the crude product 4-ethyl-5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one as a white solid (4.3 g, 88% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.52 (s, 1H), 5.55 (t, J=5.50 Hz, 1H), 4.32 (d, J=5.50 Hz, 2H), 3.64 (q, J=6.97 Hz, 2H), 1.18 (t, J=6.97 Hz, 3H) ppm.

Step B. 5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a solution of 4-ethyl-5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (3 g, 21 mmol, 1.0 equiv.) in DCM (30 mL) was added tert-butylchlorodiphenylsilane (6.5 mL, 25 mmol, 1.2 equiv.) and pyridine (1.86 mL, 23 mmol, 1.1 equiv.). The resulting mixture was stirred overnight at room temperature. The reaction mixture was quenched with water (100 mL). The resulting mixture was extracted with DCM (3×100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (SiO ₂ , 50-80% ethyl acetate/petroleum ether) to give 5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one as a white solid (4.9 g, 61% yield). LCMS (ES-API): mass calculated for C ₂₁ H ₂₇ N ₃ O ₂ Si, 381.2; observed m/z, 382.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.98 (s, 1H), 7.61-7.72 (m, 4H), 7.32-7.54 (m, 6H), 4.54 (s, 2H), 3.84 (q, J=7.34 Hz, 2H), 1.33 (t, J=7.34 Hz, 3H), 1.07 (s, 9H) ppm.

Intermediate 3: 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.

Step A. tert-Butyl 4,5-difluoro-2-iodobenzoate.
4,5-Difluoro-2-iodobenzoic acid (3 g, 11 mmol) was dissolved in THF (30 mL) then di-tert-butyl dicarbonate (4.6 g, 21 mmol) was added followed by DMAP (645 mg, 5.3 mmol). The reaction mixture was stirred under nitrogen at 50° C. overnight and then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was diluted with EtOAc and washed with brine. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-5% EtOAc in petroleum ether) to give the title compound as a yellow oil (2.9 g, yield: 79%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.77 (dd, J=10.2, 7.9 Hz, 1 H), 7.63 (dd, J=10.2, 7.9 Hz, 1 H), 1.62 (s, 9 H) ppm; ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -131.55--131.13 (m, 1 F), -136.97--136.65 (m, 1 F) ppm.

Step B. tert-Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-5-fluoro-2-iodobenzoate.
A mixture of tert-butyl 4,5-difluoro-2-iodobenzoate (3.2 g, 9.4 mmol), 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one (Intermediate 1, 2.6 g, 11.2 mmol) and Cs ₂ CO ₃ (6.1 g, 18.7 mmol) in anhydrous DMF (30 mL) was stirred at 75° C. under nitrogen for 1 h and then cooled to room temperature. The mixture was filtered through a pad of Celite® and the pad was washed with EtOAc. The filtrates were combined, washed with brine and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-40% EtOAc in petroleum ether) to give the title compound as a colorless amorphous solid (5 g, yield: 96%). ESI (MS): _mass calculated _{for C23H25FIN3O4} , _553.1 ; observed m/z, 554.1 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.16 (d, J = 7.1 Hz, 1H), 7.62 (d, J = 10.8 Hz, 1H), 7.29-7.45 (m, 5H), 4.61 (s, 2H), 4.50 (s, 2H), 3.84 (q, J = 7.2 Hz, 2H), 1.63 (s, 9H), 1.35 (t, J = 7.2 Hz, 3H) ppm; ^19F NMR (376MHz, CDCl3) δ -119.09 ( _dd , J = 10.6, 7.0 Hz, 1F) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-5-fluoro-2-iodobenzoic acid.
To a solution of tert-butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoate (5 g, 9 mmol) in DCM (50 mL) was added TFA (10 mL) slowly. The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under vacuum. The resulting residue was triturated with petroleum ether at room temperature for 30 min. The mixture was filtered and the solid was rinsed with petroleum ether. The precipitate was collected and dried in vacuum to give the title compound as a white solid (4.1 g, yield: 91%). ESI (MS): mass calculated for C ₁₉ H ₁₇ FIN ₃ O ₄ , 497.0; observed m/z, 498.0 [M+H] ⁺ . ^1H NMR (400MHz, DMSO- _d6 ) δ 8.16 (d, J = 7.3 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.28-7.43 (m, 5H), 4.60 (s, 2H), 4.57 (s, 2H), 3.74 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H) ppm; ^19F NMR (376MHz, DMSO- _d6 ) δ -119.91 (s, 1F) ppm.

Step D. 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a mixture of 3-methylbuta-1,2-dien-1-yl acetate (Intermediate 12, 280 mg, 2.2 mmol), 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-5-fluoro-2-iodobenzoic acid (1.1 g, 2.2 mmol) and Cy ₂ NMe (867 mg, 4.4 mmol) in DMF (7 mL) was added Catacxium A Pd G2 (74.2 mg, 0.11 mmol) under nitrogen. The reaction mixture was stirred under nitrogen at 90° C. overnight. The mixture was then cooled to room temperature, diluted with EtOAc and washed with brine. The organic layer was separated and the aqueous layers were combined and extracted with EtOAc. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-80% EtOAc in petroleum ether) to give the title compound as a yellow solid (240 mg, yield: 25%). ESI (MS): mass calculated for C ₂₄ H ₂₂ FN ₃ O ₄ , 435.2; m/z 436.2 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.15 (d, J = 10.5 Hz, 1H), 7.86 (d, J = 6.8 Hz, 1H), 7.30-7.45 (m, 5H), 7.19 (s, 1H), 5.37-5.39 (m, 1H), 5.18 (s, 1H), 4.62 (s, 2H), 4.53 (s, 2H), 3.87 (q, J = 7.1 Hz, 2H), 2.11 (s, 3H), 1.37 (t, J = 7.2 Hz, 3H) ppm.

Intermediate 4: 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-4-isopropyl-1-oxo-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.

Method I:
Step A. 3-Methylbut-2-en-1-yl 4,5-difluoro-2-iodobenzoate.
To a mixture of 4,5-difluoro-2-iodobenzoic acid (1.4 g, 4.9 mmol) and Cs ₂ CO ₃ (4.8 g, 14.8 mmol) in anhydrous DMF (20 mL) was added 1-bromo-3-methyl-2-butene (1.5 g, 9.9 mmol). The reaction mixture was stirred at room temperature for 18 h. The mixture was diluted with water and extracted with DCM and EtOAc. The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , gradient elution: 10-20% EtOAc in heptane) to give the desired product as a colorless oil (1.6 g, yield: 92%). ^1H NMR (400MHz, _CDCl3 ) δ 7.80 (dd, J = 7.58, 9.54 Hz, 1H), 7.73 (dd, J = 7.83, 10.76 Hz, 1H), 5.42-5.52 (m, 1H), 4.82 (d, J = 7.34 Hz, 2H), 1.80 (s, 3H), 1.78 (s, 3H) ppm.

Step B. 3-Methylbut-2-en-1-yl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoate.
To a mixture of 3-methylbut-2-en-1-yl 4,5-difluoro-2-iodobenzoate (1.6 g, 4.5 mmol), 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one (Intermediate 1, 2.1 g, 9.1 mmol) in anhydrous DMF (25 mL) was added Cs ₂ CO ₃ (2.9 g, 9.1 mmol). The reaction mixture was heated at 85° C. under nitrogen for 1 h and then cooled to room temperature. The mixture was diluted with water and the mixture was extracted with DCM and EtOAc. The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , gradient elution: 20-50% EtOAc in heptane) to give the title compound as a white solid (2.4 g, yield: 93%). LCMS (ES-API): mass calculated for C ₂₄ H ₂₅ FIN ₃ O ₄ , 565.1; observed m/z, 566.2 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.21 (d, J = 6.85 Hz, 1H), 7.73 (d, J = 11.25 Hz, 1H), 7.29-7.44 (m, 5H), 5.41-5.53 (m, 1H), 4.84 (d, J = 7.34 Hz, 2H), 4.60 (s, 2H), 4.50 (s, 2H), 3.84 (q, J = 7.22 Hz, 2H), 1.80 (s, 3H), 1.78 (d, J = 0.98 Hz, 3H), 1.34 (t, J = 7.22 Hz, 3H) ppm.

Step C. 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-4-isopropyl-1-oxo-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a mixture of 3-methylbut-2-en-1-yl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoate (4 g, 6.86 mmol, 1 equiv) in toluene (200 mL) was added (tBu ₃ P)PdG ₂ (351 mg, 0.69 mmol, 0.1 equiv), N-cyclohexyl-N-methyl-cyclohexanamine (1.60 mL, 7.54 mmol, 1.1 equiv). The reaction mixture was degassed three times with nitrogen and then heated at 80° C. under a nitrogen atmosphere for 18 h. LCMS analysis indicated approximately 18% starting material remained. The mixture was cooled to 15° C. and additional N-cyclohexyl-N-methyl-cyclohexanamine (0.72 mL, 3.43 mmol, 0.5 equiv.) and tBu ₃ PPdG ₂ (176 mg, 0.34 mmol, 0.05 equiv.) were added. The reaction mixture was degassed with nitrogen and then heated at 80° C. under nitrogen atmosphere for 16 h. The mixture was concentrated under reduced pressure and then diluted with H ₂ O (200 mL) and extracted with EtOAc (150 mL×3). The combined organic layers were washed with brine (100 mL×2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (SiO ₂ , petroleum ether/ethyl acetate=5/1 to 3/1) to give the title compound as a yellow oil (1.1 g, 35%). ESI (MS): mass calculated _{for C24H24FN3O4} , _437.2 ; found m/z, 438.5 [M+H] ⁺ . ^1H NMR (400 MHz, _CDCl3 ) δ 8.16 (d, J ₌ 6.6 Hz, 1H), 7.96 (d, J = 6.6 Hz, 1H), 7.42-7.34 (m, 5H), 7.13 (s, 1H), 4.63 (s, 2H), 4.54 (s, 2H), 3.88 (dd, J = 7.2, 14.4 Hz, 2H), 3.13-3.06 (m, 1H), 1.38 (t, J = 7.2 Hz, 3H), 1.32 (d, J = 6.8 Hz, 6H) ppm.

Method II:
To a mixture of 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (Intermediate 3, 5.9 g, 13.5 mmol) in THF (100 mL) was added Wilkinson's catalyst [RhCl(PPh ₃ ) ₃ ] (3.8 g, 4.1 mmol) at room temperature. The mixture was degassed and purged with hydrogen gas. The reaction mixture was stirred under hydrogen atmosphere (15 Psi) at room temperature for 12 hours. The mixture was concentrated. The residue was purified by silica column chromatography (elution: 0-25% EtOAc in petroleum ether) to give the title compound as a yellow solid (1.5 g, yield: 77%). ESI (MS): _mass calculated _{for C24H24FN3O4 ,} 437.2; observed m/z, 438.2 [M+H] ⁺ . ^1H NMR (400MHz, DMSO- _d6 ) δ 8.10 (d, J = 10.5 Hz, 1H), 8.01 (d, J = 7.0 Hz, 1H), 7.46 (s, 1H), 7.26-7.42 (m, 5H), 4.61 (s, 2H), 4.59 (s, 2H), 3.77 (q, J = 7.3 Hz, 2H), 3.08 (dt, J = 13.4, 6.8 Hz, 1H), 1.22-1.28 (m, 9H) ppm; ^19F NMR (376MHz, DMSO- _d6 ) δ -118.29 (br s, 1F) ppm.

Intermediate 5: 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-isobenzoyl chloride.

Step A. tert-Butyl 4,5-difluoro-2-iodobenzoate.
4,5-Difluoro-2-iodobenzoic acid (3 g, 11 mmol) was dissolved in THF (30 mL) then di-tert-butyl dicarbonate (4.6 g, 21 mmol) was added followed by DMAP (645 mg, 5.3 mmol). The reaction mixture was stirred under nitrogen at 50° C. overnight and then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was diluted with EtOAc and washed with brine. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-5% EtOAc in petroleum ether) to give the title compound as a yellow oil (2.9 g, yield: 79%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.77 (dd, J=10.2, 7.9 Hz, 1 H), 7.63 (dd, J=10.2, 7.9 Hz, 1 H), 1.62 (s, 9 H) ppm; ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -131.55--131.13 (m, 1 F), -136.97--136.65 (m, 1 F) ppm.

Step B. tert-Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-5-fluoro-2-iodobenzoate.
A mixture of tert-butyl 4,5-difluoro-2-iodobenzoate (3.2 g, 9.4 mmol), 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one (Intermediate 1, 2.6 g, 11.2 mmol) and Cs ₂ CO ₃ (6.1 g, 18.7 mmol) in anhydrous DMF (30 mL) was stirred at 75° C. under nitrogen for 1 h and then cooled to room temperature. The mixture was filtered through a pad of Celite® and the pad was washed with EtOAc. The filtrates were combined, washed with brine and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-40% EtOAc in petroleum ether) to give the title compound as a colorless amorphous solid (5 g, yield: 96%). ESI (MS): _mass calculated _{for C23H25FIN3O4} , _553.1 ; observed m/z, 554.1 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.16 (d, J = 7.1 Hz, 1H), 7.62 (d, J = 10.8 Hz, 1H), 7.29-7.45 (m, 5H), 4.61 (s, 2H), 4.50 (s, 2H), 3.84 (q, J = 7.2 Hz, 2H), 1.63 (s, 9H), 1.35 (t, J = 7.2 Hz, 3H) ppm; ^19F NMR (376MHz, CDCl3) δ -119.09 ( _dd , J = 10.6, 7.0 Hz, 1F) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-5-fluoro-2-iodobenzoic acid.
To a solution of tert-butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoate (5 g, 9 mmol) in DCM (50 mL) was added TFA (10 mL) slowly. The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under vacuum. The resulting residue was triturated with petroleum ether at room temperature for 30 min. The mixture was filtered and the solid was rinsed with petroleum ether. The precipitate was collected and dried in vacuum to give the title compound as a white solid (4.1 g, yield: 91%). ESI (MS): mass calculated for C ₁₉ H ₁₇ FIN ₃ O ₄ , 497.0; observed m/z, 498.0 [M+H] ⁺ . ^1H NMR (400MHz, DMSO- _d6 ) δ 8.16 (d, J = 7.3 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.28-7.43 (m, 5H), 4.60 (s, 2H), 4.57 (s, 2H), 3.74 (q, J = 7.2 Hz, 2H), 1.23 (t, J = 7.2 Hz, 3H) ppm; ^19F NMR (376MHz, DMSO- _d6 ) δ -119.91 (s, 1F) ppm.

Step D. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoyl chloride.
A solution of 4-( ₃ -((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluoro-2-iodobenzoic acid (3.5 g, 7 mmol) in SOCl 2 (14 mL) was heated at reflux for 15 min. The reaction mixture was cooled to room temperature and concentrated. Anhydrous toluene was added to the residue and the mixture was evaporated to give the crude product as a yellow gum (3.6 g), which was used directly in the next step without further purification.

Intermediate 6: 2-chloro-6-fluoro-N-(3-methylpent-2-en-1-yl)aniline.

Step A. Ethyl 3-methylpent-2-enoate.
To a solution of 2-butanone (52 g, 717.6 mmol) and (carbethoxymethylene)triphenylphosphorane (50 g, 143.5 mmol) in toluene (65 mL) was added benzoic acid (3.5 g, 28.7 mmol). The reaction mixture was heated to reflux for 16 h. The mixture was diluted with petroleum ether and filtered through a short pad of silica gel. The silica gel was washed with hexanes. The filtrate was concentrated under reduced pressure at 0-2° C. The residue was purified by silica column chromatography (elution: 0-10% EtOAc in petroleum ether) to give the title compound as a colorless liquid (23.3 g, crude). ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.58-5.69 (m, 1 H), 4.13 (qd, J=7.1, 4.9 Hz, 2 H), 2.62 (q, J=7.5 Hz, 1 H), 2.09-2.20 (m, 3 H), 1.86 (d, J=1.2 Hz, 1 H), 1.24-1.28 (m, 3 H), 1.01-1.09 (m, 3 H) ppm.

Step B. 3-Methylpent-2-en-1-ol.
To a 1M solution of DIBAL-H (118 mL, 118 mmol) in toluene at −78° C., under nitrogen, was added dropwise a solution of ethyl 3-methylpent-2-enoate (20 g, crude) in toluene (40 mL). The reaction mixture was stirred at −78° C. for 2 h. The mixture was warmed to room temperature and slowly poured into saturated aqueous potassium sodium tartrate at 0° C. The mixture was stirred for 2 h and filtered through a short pad of Celite®. The pad was washed with DCM/EtOAc (v/v, 3/1) and the filtrate was extracted with DCM. The organic extract was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (elution: 0-100% DCM in petroleum ether, then 0-30% EtOAc in DCM) to give the title compound as a colorless liquid (7 g, 2 steps yield: 57%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.35-5.46 (m, 1 H), 4.11-4.21 (m, 2 H), 2.02-2.13 (m, 2 H), 1.67-1.76 (m, 3 H), 0.98-1.06 (m, 3 H) ppm.

Step C. 3-Methylpent-2-enal.
To a solution of 3-methylpent-2-en-1-ol (2 g, 20.0 mmol) in DCM (20 mL) was added Dess-Martin periodinane (10 g, 24.0 mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was filtered through a short pad of Celite®. The pad was washed with DCM. The combined filtrate was washed with saturated aqueous NaHCO ₃ solution. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure at 0-2° C. The crude product was purified by silica column chromatography (elution: DCM) to give the title compound as a colorless liquid (1.5 g, yield: 77%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.91-10.04 (m, 1 H), 5.78-5.90 (m, 1 H), 2.58 (q, J=7.6 Hz, 1 H), 2.23 (d, J=7.3 Hz, 1 H), 2.16 (s, 2 H), 1.96 (d, J=1.1 Hz, 1 H), 1.16 (t, J=7.6 Hz, 1 H), 1.09 (t, J=7.4 Hz, 2 H) ppm.

Step D. N-(2-chloro-6-fluorophenyl)-3-methylpent-2-en-1-imine.
To a mixture of 2-chloro-6-fluoroaniline (1.2 g, 8.2 mmol) and 3-methylpent-2-enal (0.97 g, 9.9 mmol) in DCM (18 mL) at 0° C. under nitrogen, triethylamine (4.6 mL, 33 mmol) was added followed by the dropwise addition of a 1 M solution of TiCl ₄ (5 mL, 5 mmol) in DCM. The resulting mixture was stirred at 0° C. for 1 h, then allowed to warm to room temperature and stirred for 4 h. The mixture was poured into saturated aqueous NH ₄ Cl. The mixture became cloudy and was filtered through a pad of Celite®. The pad was washed with EtOAc. The combined filtrate was diluted with DCM and water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residual product was purified by silica gel column chromatography (gradient elution: 0-5% DCM in petroleum ether) to afford the title compound as a pale yellow oil (1.3 g, yield: 70%).

Step E. 2-Chloro-6-fluoro-N-(3-methylpent-2-en-1-yl)aniline.
To a solution of N-(2-chloro-6-fluorophenyl)-3-methylpent-2-en-1-imine (1.3 g, 5.76 mmol) in MeOH (20 mL) was added NaBH ₄ (218 mg, 5.8 mmol) and after 1 h another batch of NaBH ₄ (218 mg, 5.8 mmol) was added. A total of NaBH ₄ (1.1 g, 29 mmol) was added. The reaction mixture was stirred at room temperature overnight. The mixture was concentrated, then diluted with water and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by Combi-Flash column chromatography on silica gel (eluent: 0-5% DCM in petroleum ether) to give the title compound as a yellow oil (430 mg, yield: 33%). ^1H NMR (400MHz, _CDCl3 ) δ 6.95-7.02 (m, 1H), 6.84 (ddd, J=12.2, 8.3, 1.3Hz, 1H), 6.52-6.63 (m, 1H), 5.17-5.28 (m, 1H), 3.85 (d, J=5.6Hz, 2H), 3.73 (s, 1H), 1.91-2.07 (m, 2H), 1.58-1.68 (m, 3H), 0.89-0.96 (m, 3H).

Intermediate 7: 5-chloro-3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-amine.

Step A. 5-Chloro-3-methyl-4-nitro-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole.
To a solution of 3-methyl-4-nitropyrazole (2 g, 15.7 mmol) in EtOAc (20 mL) was added DHP (2 g, 23.6 mmol) and TsOH.H ₂ O (150 mg, 0.79 mmol) at room temperature. The mixture was stirred at room temperature overnight. Et ₃ N (0.4 mL) was added and the mixture was washed with brine. The organic layer was then separated, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was dissolved in THF (45 mL) and the temperature was lowered to −78° C. A solution of LiHMDS (10.6 mL, 13.8 mmol) in THF (1 M) was added to the mixture under nitrogen. After 45 min at −78° C., a solution of hexachloroethane (8.9 g, 37.8 mmol) in THF (20 mL) was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred overnight. The mixture was poured into saturated aqueous NH ₄ Cl and extracted with EtOAc. The organic phase was separated, washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-40% EtOAc in petroleum ether) to give the title compound as a white solid (1.8 g, yield: 58%). ^1H NMR (400MHz, _CDCl3 ) δ 5.52 (dd, J=10.0, 2.7Hz, 1H), 4.07-4.15 (m, 1H), 3.70 (td, J=11.3, 2.8Hz, 1H), 2.57 (s, 3H), 2.37-2.47 (m, 1H), 2.11-2.19 (m, 1H), 1.86-1.90 (m, 1H), 1.72-1.75 (m, 1H), 1.64 (d, J=2.0Hz, 1H), 1.53 (d, J=6.6Hz, 1H) ppm.

Step B. 5-Chloro-3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-amine.
To a mixture of 5-chloro-3-methyl-4-nitro-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole (100 mg, 0.4 mmol) in MeOH/THF/H ₂ O (v/v/v, 1/1/1, 3 mL) was added iron powder (114 mg, 2.0 mmol) and NH ₄ Cl (109 mg, 2.0 mmol). The mixture was stirred at 70° C. for 1.5 h. The mixture was cooled to room temperature and filtered through a pad of Celite®. The pad was washed with EtOAc. The combined filtrate was washed with saturated aqueous NaHCO _3. The organic layer was separated and the aqueous layer was extracted with EtOAc. The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-50% EtOAc in petroleum ether) to give the title compound as a yellow oil (70 mg, yield: 79%). ESI (MS): mass calculated for C ₉ H ₁₄ ClN ₃ O, 215.1; observed m/z, 216.1 [M+H] ⁺ .

Intermediate 8: 3-(2-((tert-butyldiphenylsilyl)oxy)ethoxy)-2-chloroaniline.

Step A. tert-Butyl(2-(2-chloro-3-nitrophenoxy)ethoxy)diphenylsilane.
To a mixture of 2-chloro-3-nitrophenol (200 mg, 1.2 mmol), 2-((tert-butyldiphenylsilyl)oxy)ethan-1-ol (554 mg, 1.8 mmol) and PPh ₃ (453 mg, 1.7 mmol) in THF (10 mL) at 0° C. under nitrogen was added DEAD (281 mg, 161 mmol). The mixture was allowed to warm to room temperature and stirred at room temperature for 12 h. Saturated aqueous NH ₄ Cl was added and the mixture was extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na 2 SO 4 , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-10% EtOAc in petroleum ether) to give the title compound as a yellow oil (240 mg, yield: 46%). ^1H NMR (400MHz, _CDCl3 ) δ 7.72 (dd, J=7.8, 1.5Hz, 4H), 7.35-7.49 (m, 7H), 7.30 (t, J=8.2Hz, 1H), 7.12 (dd, J=8.3, 1.2Hz, 1H), 4.20-4.25 (m, 2H), 4.06 (t, J=4.9Hz, 2H), 1.06 (s, 9H) ppm.

Step B. 3-(2-((tert-butyldiphenylsilyl)oxy)ethoxy)-2-chloroaniline.
To a mixture of tert-butyl(2-(2-chloro-3-nitrophenoxy)ethoxy)diphenylsilane (220 mg, 0.5 mmol), NH ₄ Cl (258 mg, 4.8 mmol) in THF (3 mL), MeOH (3 mL) and H2O (3 mL) was added iron powder (269 mg, 4.8 mmol). The reaction mixture was stirred at 70° C. for 2 h. The mixture was cooled to room temperature, diluted with EtOAc and filtered through a pad of Celite®. The Celite® was washed with EtOAc. The combined filtrate was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-11% EtOAc in petroleum ether) to give the title compound as a yellow solid (192 mg, yield: 92%). ESI (MS): _mass calculated _{for C24H28ClNO2Si} , 425.2; observed m/z, 426.1 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 7.75 (dd, J = 7.9, 1.6Hz, 4H), 7.35-7.48 (m, 6H), 6.97 (t, J = 8.1Hz, 1H), 6.42 (dd, J = 8.2, 1.1Hz, 1H), 6.33 (dd, J = 8.2, 1.1Hz, 1H), 4.13-4.17 (m, 2H), 4.07-4.13 (m, 2H), 4.01-4.06 (m, 2H), 1.06 (s, 9H) ppm.

Intermediate 9: 5-((benzyloxy)methyl)-4-ethyl-2-(7-methyl-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.

Step A. tert-Butyl 2-bromo-4-fluoro-5-methylbenzoate.
To a solution of 2-bromo-4-fluoro-5-methylbenzoic acid (1 g, 4.3 mmol) in THF (10 mL) was added (Boc) ₂ O (1.9 g, 8.6 mmol) followed by DMAP (262 mg, 2.1 mmol). The reaction mixture turned orange and was stirred at 50° C. overnight under nitrogen. The mixture was cooled to room temperature, diluted with EtOAc and washed with brine. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (elution: 0-3% EtOAc in petroleum ether) to give the title compound as a colorless oil (900 mg, yield: 72%). ^1H NMR (400 MHz, _CDCl3 ) δ 7.57 (d, J = 8.1 Hz, 1H), 7.24 (d, J = 4.6 Hz, 1H), 2.22 (d, J = 1.5 Hz, 3H), 1.58 (s, 9H) ppm.
Step B. tert-Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-2-bromo-5-methylbenzoate.

A mixture of tert-butyl 2-bromo-4-fluoro-5-methylbenzoate (750 mg, 2.6 mmol), 5-((benzyloxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (800 mg, 3.4 mmol) and Cs ₂ CO ₃ (1.7 g, 5.2 mmol) in DMF (8 mL) was stirred at 90° C. for 16 h. The reaction was quenched by addition of saturated aqueous NH ₄ Cl. The mixture was extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by Combiflash chromatography (SiO ₂ , eluent: 0-22% EtOAc in petroleum ether) to give the title compound as a colorless gum (1 g, yield: 71%). ESI (MS): mass calculated for _C24H28BrN3O4 , _501.1 ; m/z found, 502.1 [M+H] ⁺ . ^1H NMR ( ₄₀₀ MHz, _CDCl3 ) δ 7.64 (d, J ₌ 6.1 Hz, 2H), 7.33-7.43 (m, 5H), 4.61 (s, 2H), 4.50 (s, 2H), 3.85 (q, J=7.3 Hz, 2H), 2.31 (s, 3H), 1.62 (s, 9H), 1.36 (t, J=7.2 Hz, 3H) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-2-bromo-5-methylbenzoic acid.
To a mixture of tert-butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-bromo-5-methylbenzoate (500 mg, 0.90 mmol) in DCM (5 mL) was added TFA (1 mL). The mixture was stirred at room temperature for 12 h. The mixture was concentrated. The residue was dissolved in DCM and petroleum ether was added slowly. The mixture was stirred at room temperature for 30 min. The mixture was filtered and the precipitate was rinsed with petroleum ether. The solid was collected and dried in vacuum to give the title compound as _a white solid ( _{360 mg} , yield: 86%). ESI (MS): mass calculated for _C20H20BrN3O4 , 445.1; observed m/z, 446.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.77 (s, 1 H), 7.70 (s, 1 H), 7.29-7.41 (m, 5 H), 4.59 (s, 2 H), 4.56 (s, 2 H), 3.74 (q, J=7.0 Hz, 2 H), 2.24 (s, 3 H), 1.23 (t, J=7.2 Hz, 3 H) ppm.

Step D. 5-((benzyloxy)methyl)-4-ethyl-2-(7-methyl-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a mixture of 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-bromo-5-methylbenzoic acid (560 mg, 1.26 mmol), 3-methylbut-1,2-dien-1-yl acetate (Intermediate 12, 1.58 g, 12.5 mmol), AcOK (369 mg, 3.76 mmol) and TBAB (809 mg, 2.51 mmol) in DMF (3.9 mL) was added Pd(OAc) ₂ (141 mg, 0.63 mmol) under nitrogen. The reaction mixture was stirred at 90° C. overnight under nitrogen. The mixture was cooled to room temperature, diluted with EtOAc and washed with brine. The organic layer was separated and the aqueous layer was extracted with EtOAc. The combined organic layers were combined, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-70% EtOAc in petroleum ether) to give the title compound as a yellow solid (410 mg, yield: 73%). ESI (MS): mass calculated for C ₂₅ H ₂₅ N ₃ O ₄ , 431.2; observed m/z, 432.1 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.28 (s, 1H), 7.59 (s, 1H), 7.31-7.46 (m, 5H), 7.17 (s, 1H), 5.30-5.37 (m, 1H), 5.15 (s, 1H), 4.63 (s, 2H), 4.52 (s, 2H), 3.87 (q, J = 7.1 Hz, 2H), 2.46 (s, 3H), 2.10 (s, 3H), 1.38 (t, J = 7.2 Hz, 3H) ppm.

Intermediate 10: Isopropyl 6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-chloro-5-fluoronicotinate.

Step A. 2,6-Dichloro-5-fluoronicotinoyl chloride.
To a solution of 2,6-dichloro-5-fluoronicotinic acid (20 g, 95 mmol) in THF (200 mL) at 0° C. was added (COCl) ₂ (12.7 g, 10.0 mmol) and DMF (69.6 mg, 0.952 mmol) dropwise. The mixture was stirred at 0° C. for 30 min, then warmed to 25° C. and stirred for 1 h. The reaction mixture was concentrated under reduced pressure to give the desired product (21.7 g, crude) as a colorless oil which was used without further purification.

Step B. Isopropyl 2,6-dichloro-5-fluoronicotinate.
To a mixture of propan-2-ol (8.56 g, 142 mmol, 10.9 mL) and pyridine (9.02 g, 114 mmol) in THF (200 mL) was added a solution of 2,6-dichloro-5-fluoronicotinoyl chloride (21.7 g, 96.0 mmol) in THF (50 mL) at 0° C. The mixture was stirred at 25° C. for 1 h. The mixture was poured into water (300 mL). The aqueous phase was extracted with ethyl acetate (300 mL). The combined organic phase was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/1 to 10:1) to give the title compound (21 g, 86.82% yield). MS (ESI): mass calculated _{for C9H8Cl2FNO2} , _250.1 ; m/z found, 252.0 [M+H]+. ^1H NMR (400 MHz, _CDCl3 ) δ 7.97-7.95 (d, J ₌ 7.2 Hz, 1H), 5.32-5.25 (m, 1H), 1.58-1.39 (m, 6H) ppm.

Step C. Isopropyl 6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-2-chloro-5-fluoronicotinate.
To a mixture of isopropyl 2,6-dichloro-5-fluoronicotinate (4 g, 15.87 mmol) in DMSO (40 mL) was added 3-((benzyloxy)methyl)-4-ethyl-1H-1,2,4-triazol-5(4H)-one (3.89 g, 16.66 mmol) and K ₂ CO ₃ (3.29 g, 23.80 mmol). The mixture was stirred at 80° C. for 3 h. LCMS showed the starting material was consumed and the desired material was detected. The mixture was diluted with H ₂ O (30 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (100 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0 to 1:1) to give the title compound (5.7 g, 79.86% yield). MS (ESI): mass calculated for C ₂₁ H ₂₂ ClFN ₄ O ₄ , 448.1; observed m/z, 449.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.10 (d, J=8.8 Hz, 1H), 7.43-7.31 (m, 5H), 5.30 (td, J=6.3, 12.5 Hz, 1H), 4.61 (s, 2H), 4.54 (s, 2H), 3.85 (q, J=7.2 Hz, 2H), 1.41 (d, J=6.2 Hz, 6H), 1.37-1.31 (m, 3H) ppm.

Intermediate 11: 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-3-hydroxy-4-isopropyl-1-oxoisochroman-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.

Step A. Methyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-2-bromo-5-fluorobenzoate.
To a flask containing methyl 2-bromo-4,5-difluorobenzoate (100.0 g, 398 mmol), 5-((benzyloxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (Intermediate 1, 113.5 g, 508 mmol) and K ₂ CO ₃ (100.0 g, 724 mmol) was added anhydrous DMF (1000 mL). The reaction mixture was heated at 50° C. under nitrogen for 16 hours, after which additional 5-((benzyloxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (11 g, 51 mmol) was added. The reaction mixture continued to stir at 50° C. The mixture was cooled to room temperature and stirred for 10 minutes. Water (1000 mL) was added dropwise and the mixture was stirred at room temperature for 2 hours. The precipitate was collected by filtration and dried to give the crude product (190 g). The product was stirred in DMF (500 mL) for 30 min, then water (500 mL) was added. The mixture was stirred for 2 h. The precipitate was collected by filtration and dried to give the title compound (180 g, yield: 90%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.95 (d, J = 6.7 Hz, 1H), 7.73 (d, J = 10.7 Hz, 1H), 7.44-7.27 (m, 5 H), 4.60 (s, 2 H), 4.50 (s, 2 H), 3.95 (s, 3H), 3.84 (q, J = 7.2 Hz, 2 H), 1.34 (t, J = 7.2 Hz, 3 H) ppm.

Step B. 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-3-hydroxy-4-isopropyl-1-oxoisochroman-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one.
To a mixture of methyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-2-bromo-5-fluorobenzoate (30 g, 65.9 mmol), Xantphos (4.02 g, 6.6 mmol), [Pd(allyl)Cl] ₂ (1.38 g, 3.77 mmol), Cs ₂ CO ₃ (42.6 g, 130 mmol) in dimethylacetamide (300 mL) was added 3-methylbutanal (41.4 mL, 386 mmol) slowly. The reaction mixture was heated at 80° C. under nitrogen for 22 hours. The mixture was filtered and quenched with aqueous NH ₄ Cl until the “pH” was 7-8. The mixture was extracted with ethyl acetate (2×2000 mL). The combined organic extracts were concentrated. The residue was purified by column chromatography (SiO ₂ , gradient elution: 1-33% ethyl acetate in petroleum ether) to afford the title compound as an oil (61.7 g, yield: 56%). ^1H NMR (300MHz, _CDCl3 ) δ 7.93 (d, J = 10.4 Hz, 1H), 7.55 (d, J = 6.7 Hz, 1H), 7.44-7.28 (m, 5H), 5.92 (s, 1H), 4.61 (s, 2H), 4.51 (s, 2H), 4.37 (br s, 1H), 3.85 (q, J = 7.2 Hz, 2H), 2.80 (d, J = 7.1 Hz, 1H), 1.92 (spt, J = 6.8 Hz, 1H), 1.35 (t, J = 7.2 Hz, 3H), 1.05 (d, J = 6.8 Hz, 3H), 0.93 (d, J = 6.8 Hz, 3H) ppm.

Intermediate 12: 3-Methylbut-1,2-dien-1-yl acetate.

Step A. 2-Methylbut-3-yn-2-yl acetate.
2-Methyl-3-butyn-2-ol (30 g, 357 mmol) was added dropwise to a mixture of Mg(ClO ₄ ) ₂ (796 mg, 3.6 mmol) in acetic anhydride (38 g, 371 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 10 min, then warmed to room temperature and stirred overnight. The reaction mixture was diluted with DCM and then washed with saturated aqueous NaHCO ₃ and saturated aqueous Na ₂ CO _3. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and concentrated at 0° C. The residue was purified by silica column chromatography (elution: DCM) to give the title compound as a pale yellow oil (35.8 g, yield: 80%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.54 (s, 1 H), 2.03 (s, 3 H), 1.67 (s, 6 H) ppm.

Step B. 3-Methylbut-1,2-dien-1-yl acetate.
To a solution of 2-methylbut-3-yn-2-yl acetate (2.5 g, 20 mmol) in DCM (20 mL) was added AgBF ₄ (117 mg, 0.6 mmol) under nitrogen. The resulting colorless solution was stirred at 35° C. under nitrogen for 2 h until the mixture became a black solution. The mixture was washed with aqueous ammonia (10%). The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by silica column chromatography (gradient elution: 0-3% EtOAc in petroleum ether) to give the title compound as a yellow oil (650 mg, yield: 26%). ^1H NMR (400 MHz, _CDCl3 ) δ 7.20 (dt, J = 4.1, 2.0 Hz, 1H), 2.11 (s, 3H), 1.81 (d, J = 2.0 Hz, 6H) ppm.

Example 22 of International Application No. IB2020/053601 (published as WO 2020/212897 on October 22, 2020) discloses 6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one (compound 22).

Step A. 6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one.
To a mixture of 5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-3-hydroxy-4-isopropyl-1-oxoisochroman-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (intermediate 11, 56 g, 123 mmol) in AcOH (160 mL) was added o-toluidine (14.8 g, 138 mmol). The reaction mixture was heated at 80° C. for 16 h. The mixture was concentrated and then the “pH” was adjusted to 7-8 with aqueous NaHCO ₃ solution. The mixture was extracted with ethyl acetate (160 mL×2). The combined organic extracts were concentrated. The residue was purified by flash chromatography (SiO ₂ , 0-20% ethyl acetate in DCM) to give the title compound as an oil (32.5 g, yield: 50%). MS (ESI): mass calculated _{for C31H31FN4O3 , 526.2} ; observed m/z, 527.4 [M+H] ⁺ . ^1H NMR (300MHz, _CDCl3 ) δ 8.33 (d, J = 10.9 Hz, 1H), 8.07 (d, J = 6.8 Hz, 1H), 7.44-7.27 (m, 9H), 6.84 (s, 1H), 4.64 (s, 2H), 4.55 (s, 2H), 3.89 (q, J = 7.2 Hz, 2H), 3.23 (spt, J = 6.8 Hz, 1H), 2.16 (s, 3H), 1.39 (t, J = 7.2 Hz, 3H), 1.31 (dd, J = 6.8, 2.1 Hz, 6H) ppm.

Step B. 6-(4-Ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one.
To a stirred solution of 6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4 triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one (26.5 g, 50.3 mmol) in DCM (230 mL) at −78° C. under nitrogen was added a 1 M solution of BCl ₃ (290 mL, 290 mL) in DCM. The reaction mixture was stirred at 15° C. for 0.5 h. The reaction was quenched to −20° C. with MeOH (100 mL) at −78° C. The mixture was partitioned between water and DCM. The organic layer was separated and the aqueous layer was extracted with DCM (2×110 mL). The combined organic extracts were washed with brine (30 mL x 2), dried over _{anhydrous Na2SO4} , filtered and concentrated to give the crude product (27.5 g). The product was triturated with methyl ethyl ketone (82 mL) and heptane (290 mL) to give the pure product (17.5 g). This was recrystallized with ethanol and water to give the title compound as a white solid ( ₁₆ g, yield: 73%). MS (ESI): mass calculated _{for C24H25FN4O3} , _436.2 ; m/z found, 437.2 [M+H] ⁺ . ^1H NMR (400MHz, _CDCl3 ) δ 8.33 (d, J = 11.2, 1H), 8.08 (d, J = 6.8, 1H), 7.39-7.33 (m, 3H), 7.28 (s, 1H), 6.85 (s, 1H), 4.69 (br s, 2H), 3.94 (q, J = 7.11Hz, 2H), 3.27 (td, J = 13.66, 6.82Hz, 1H), 2.32 (br s, 1H), 2.17 (s, 3H), 1.45 (t, J = 7.11Hz, 3H), 1.32 (dd, J = 6.82, 1.83Hz, 6H) ppm.

It is noted that the compounds of formula (Z) described herein are described in International Application No. IB2020/053601 (published as WO 2020/212897 on October 22, 2020), the entirety of which is incorporated herein by reference for all purposes.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other anti-neoplastic agent.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a hypomethylating agent.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a hypomethylating agent.

According to an embodiment, the hypomethylating agent is azacitidine, decitabine, or a pharma- ceutically acceptable salt or solvate thereof.

In certain embodiments, the hypomethylating agent is azacitidine, or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof and a BCL-2 inhibitor.

In some embodiments, a combination therapy is provided that comprises a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof and a BCL-2 inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof and venetoclax or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A, or a pharma- ceutically acceptable salt or solvate thereof, and venetoclax, or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A1 and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A2 and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A3 and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4 and venetoclax or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and azacitidine or a pharma-ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutical acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is decitabine or a pharma- ceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DNA intercalating agent.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DNA intercalating agent.

According to an embodiment, the DNA intercalating agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is an anthracycline.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a pyrimidine analog.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a pyrimidine analog.

According to an embodiment, the pyrimidine analog is cytarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is cytarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a purine analog.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a purine analog.

According to an embodiment, the purine analog is fludarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an IDH inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an isocitrate dehydrogenase-1 inhibitor (e.g., ivosidenib).

According to an embodiment, the IDH inhibitor is ivosidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an isocitrate dehydrogenase-2 inhibitor (e.g., enadidenib).

According to an embodiment, the IDH inhibitor is enadidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is enadidenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an immunomodulatory antineoplastic agent.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an immunomodulatory antineoplastic agent that is a PD-1 inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an immunomodulatory antineoplastic agent.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an immunomodulatory antineoplastic agent that is a PD-1 inhibitor.

According to embodiments, the immunomodulatory antineoplastic agent is nivolumab, atezolizumab, pembrolizumab, thalidomide, lenalidomide, pomalidomide, killed Mycobacterium bovis (BCG), or levamisole.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lenalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor.

According to an embodiment, the DHODH inhibitor is a compound of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z) or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z) or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a kinase inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a kinase inhibitor.

According to an embodiment, the kinase inhibitor is a serine and/or tyrosine kinase inhibitor.

According to an embodiment, the kinase inhibitor is an inhibitor of FLT3 and/or BTK.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an FLT3 inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is an FLT3 inhibitor.

According to embodiments, the FLT3 inhibitor is sorafenib, sunitinib, midostaurin (PKC412), lestaurtinib (CEP-701), tanzutinib (MLN518), quizartinib (AC220), gilteritinib (ASP2215), or KW-2449.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sorafenib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is midostaurin.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is tandutinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is AC220.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a BTK inhibitor.

According to an embodiment, the BTK inhibitor is ibrutinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutical acceptable salt or solvate thereof, venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is a CD20 inhibitor.

According to an embodiment, the CD20 inhibitor is an anti-CD20 antibody, in particular obinutuzumab (GA101).

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes a menin-MLL inhibitor of formula (I) or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, where the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A or a pharma- ceutically acceptable salt or solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A1, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A2, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A3, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A4-a or a solvate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A4-b or a hydrate thereof, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy is provided that includes compound A4, venetoclax or a pharma- ceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent, wherein the at least one other antineoplastic agent is GA101.

All possible combinations of the above embodiments are considered to be within the scope of the present invention.

In some embodiments, methods are provided for treating a subject diagnosed with a hematopoietic disorder. The invention relates to novel methods, for example, comprising administering to a subject a therapeutically effective amount of a menin-MLL inhibitor as described herein, a therapeutically effective amount of a BCL-2 inhibitor, and, optionally, a therapeutically effective amount of at least one other anti-neoplastic agent.

An additional embodiment of the present invention relates to a method described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is orally administered to a subject.

Additional embodiments of the invention relate to methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to a subject at a dose of about 1 mg/kg to about 50 mg/kg.

Additional embodiments of the invention relate to methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to a subject at a dose of about 2.5 mg/kg to about 25 mg/kg.

Additional embodiments of the invention relate to methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to the subject at a dose of about 7.5 mg/kg to about 12.5 mg/kg.

Additional embodiments of the invention relate to methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to the subject at a dose of about 8 mg/kg to about 10 mg/kg.

Additional embodiments of the invention relate to methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to a subject at a dose of about 0.1 mg to about 5 mg.

Additional embodiments of the invention relate to methods described herein, in which a BCL-2 inhibitor is orally administered to a subject.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered to the subject at a dose of about 1 mg/kg to about 50 mg/kg.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered to the subject at a dose of about 2.5 mg/kg to about 25 mg/kg.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered to the subject at a dose of about 7.5 mg/kg to about 12.5 mg/kg.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered to the subject at a dose of about 8 mg/kg to about 10 mg/kg.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered to the subject at a dose of about 0.1 mg to about 5 mg.

Additional embodiments of the invention relate to methods described herein, wherein at least one other anti-neoplastic agent is administered intravenously or subcutaneously to the subject.

Additional embodiments of the present invention relate to the methods described herein, wherein at least one other anti-neoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 10 mg/m ² to about 250 mg/m ² .

Additional embodiments of the present invention relate to the methods described herein, wherein at least one other anti-neoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 50 mg/m ² to about 150 mg/m ² .

Additional embodiments of the present invention relate to the methods described herein, wherein at least one other anti-neoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 60 mg/m ² to about 100 mg/m ² .

An additional embodiment of the present invention relates to the methods described herein, wherein at least one other anti-neoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 75 mg/ ^m2 .

Additional embodiments of the invention relate to methods described herein, wherein at least one other anti-neoplastic agent is orally administered to the subject at a dose of about 1 mg to about 500 mg.

Additional embodiments of the invention include a method for treating a subject diagnosed with cancer (e.g., the cancer is a leukemia, such as myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small lymphocytic lymphoma (SLL), or chronic lymphocytic leukemia (CLL)), comprising:
a therapeutically effective amount of a menin-MLL inhibitor of formula (I), or a tautomer or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof;
administering to a subject a therapeutically effective amount of a DHODH inhibitor of formula (Z), or a pharma- ceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof;
For example, methods include administering to a subject a menin-MLL inhibitor and a DHODH inhibitor according to their administration schedules for a period of time, e.g., (i) administered simultaneously or sequentially in any order on the same day within a period of time (e.g., a 21 day period, or a 28 day period, or a 3 month period, or a 6 month period, or a 1 year period, etc.), and/or (ii) administered on different days within a period of time.

Additional embodiments of the invention include a method for treating a subject diagnosed with cancer (e.g., the cancer is a leukemia, such as myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small lymphocytic lymphoma (SLL), or chronic lymphocytic leukemia (CLL)), comprising:
administering to a subject a therapeutically effective amount of a menin-MLL inhibitor which is Compound A or a pharma- ceutically acceptable salt or solvate thereof (e.g., Compound A1, or Compound A2, or Compound A3, or Compound A4-a, or a solvate thereof, or Compound A4-b, or a hydrate thereof, or Compound A4), and a therapeutically effective amount of a DHODH inhibitor which is Compound 22, or a pharma- ceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof (e.g., 6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one),
For example, methods include administering to a subject a menin-MLL inhibitor and a DHODH inhibitor according to their administration schedules for a period of time, e.g., (i) administered simultaneously or sequentially in any order on the same day within a period of time (e.g., a 21 day period, or a 28 day period, or a 3 month period, or a 6 month period, or a 1 year period, etc.), and/or (ii) administered on different days within a period of time.

An additional embodiment of the invention relates to a method described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered daily to a subject.

An additional embodiment of the invention relates to the methods described herein, wherein a compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof is administered to a subject daily for at least seven days.

Additional embodiments of the invention relate to methods described herein, in which the BCL-2 inhibitor is administered daily to the subject.

Additional embodiments of the invention relate to methods described herein, in which a BCL-2 inhibitor is administered to a subject daily for at least seven days.

Additional embodiments of the invention relate to methods described herein, in which at least one other anti-neoplastic agent is administered to the subject daily.

Additional embodiments of the invention relate to methods described herein, wherein at least one other anti-neoplastic agent is administered to the subject daily for at least 21 days.

Optimal dosages of any of the therapeutic compounds described herein to be administered can be readily determined and will vary with the particular compound used, the mode of administration, the strength of the preparation, and the progression of the disease, syndrome, condition, or disorder. In addition, factors related to the particular subject being treated, such as the subject's sex, age, weight, diet, and time of administration, may necessitate adjustment of the dosage to achieve appropriate therapeutic levels and the desired therapeutic effect.

The above dosage amounts are therefore exemplary of the average case. There can, of course, be individual instances in which higher or lower dosage ranges are merited, and such are within the scope of this invention.

The therapeutic compounds described herein can be administered in any of the compositions and administration regimens described above, or by compositions and administration regimens established in the art, whenever the therapeutic compounds described herein are administered to a subject in need thereof.

The therapeutic compounds described herein can be administered to a subject simultaneously or sequentially.

When administered sequentially, the menin-MLL inhibitor of formula (I) may be administered first. When administered simultaneously, the combination can be administered either in the same pharmaceutical composition or in different pharmaceutical compositions. For example, the menin-MLL inhibitor of formula (I) can be administered before, simultaneously with, or after administration of the BCL-2 inhibitor. For example, the BCL-2 inhibitor can be administered before, simultaneously with, or after administration of any other anti-neoplastic agent. Adjunctive therapy, i.e., therapy in which one or two agents are used as a primary treatment and other agents are used to support the primary treatment, is also an embodiment of the present invention.

One embodiment of the present invention relates to a therapeutically effective amount of a menin-MLL inhibitor, including compounds of formula (I) and pharma- ceutically acceptable salts and solvates thereof, or any subgroup thereof as described in any of the embodiments, for use in combination with a therapeutically effective amount of a BCL-2 inhibitor and, optionally, a therapeutically effective amount of at least one other anti-neoplastic agent. In certain embodiments, said therapeutically effective amounts are administered in separate dosage forms when used in treating a subject diagnosed with a hematopoietic disorder.

An embodiment of the present invention relates to a pharmaceutical product comprising a menin-MLL inhibitor of formula (I) and its pharma- ceutical acceptable salts and solvates, or any subgroup thereof as described in any of the embodiments, and a BCL-2 inhibitor, and optionally at least one other antineoplastic agent, as a combined preparation for simultaneous, separate or sequential use in the treatment of a subject diagnosed with a hematopoietic disorder.

The following examples are provided to illustrate some of the concepts described within this disclosure. The examples are to be considered as providing embodiments, but should not be considered as limiting the more general embodiments described herein.

EXAMPLES General Synthetic Scheme Venetoclax Venetoclax is commercially available.

Azacitidine Azacitidine is commercially available.

Compounds of Formula (I) In this section, and in all other sections, unless the context indicates otherwise, references to formula (I) also include all other subgroups and embodiments thereof defined herein.

The general preparation of some representative examples of compounds of formula (I) is described below, which in certain examples are typically prepared from starting materials that are either commercially available or prepared by standard synthetic processes commonly used by those skilled in the art of organic chemistry. The following schemes are merely illustrative of examples of the present invention and are not intended to limit the present invention in any way.

Alternatively, compounds of formula (I) may also be prepared by analogous reaction protocols as described in the general scheme below, in combination with standard synthetic processes commonly used by those skilled in the art.

Those skilled in the art will appreciate that in the reactions depicted in the schemes, although this is not always explicitly shown, it may be necessary to protect reactive functional groups (e.g., hydroxy, amino, or carboxy groups) if these are desired in the final product to avoid their undesired participation in the reaction. In general, conventional protecting groups (PG) can be used in accordance with standard procedures. The protecting groups can be removed at a subsequent convenient stage using methods well known in the art.

Those skilled in the art will appreciate that in the reactions described in the schemes, it may be advisable or necessary to carry out the reactions under an inert atmosphere, such as, for example, under an atmosphere of _N2 gas.

It will be apparent to one skilled in the art that it may be necessary to cool the reaction mixture prior to carrying out the reaction (e.g., quenching, column chromatography, extraction, etc., a series of operations required to isolate and purify the products of a chemical reaction).

Those skilled in the art will appreciate that heating the reaction mixture under stirring may enhance the reaction outcome. In some reactions, microwave heating may be used in place of conventional heating to reduce the overall reaction time.

Those skilled in the art will appreciate that the alternative series of chemical reactions shown in the following schemes may also lead to the desired compounds of formula (I).

Those skilled in the art will appreciate that the intermediates and final compounds depicted in the following schemes may be further functionalized according to methods well known to those skilled in the art. The intermediates and compounds described herein may be isolated in free form or as a salt or solvate thereof. The intermediates and compounds described herein may be synthesized in the form of mixtures of tautomeric and stereoisomeric forms, which may be separated from one another according to resolution procedures known in the art.

All abbreviations used in the general scheme for formula (I) are as defined in Table 1B in the Examples part below. Variables are as defined within ranges or as specifically defined in the general scheme.

Part A) Schemes 1a, 1b, 1c, 2a, 2b and 3

スキーム１ａ、１ｂ及び１ｃでは、以下の反応条件が適用される。
工程１：適切な温度（例えば、－７０℃など）で、適切な塩基（例えば、ＴＭＥＤＡなど）及び適切な有機金属試薬（例えば、イソプロピルマグネシウムブロミドなど）の存在下、適切な溶媒（例えば、ＴＨＦなど）中、
工程２：適切な温度（例えば、０℃～室温など）で、適切な酸化試薬（例えば、ＤＭＰなど）の存在下、適切な溶媒（例えば、ＤＣＭなど）中、
工程３：適切な温度（例えば、－２０℃～室温など）で、適切な有機金属試薬（例えばイソプロピルマグネシウムブロミドなど）の存在下、適切な溶媒（例えば、ＴＨＦなど）中、
工程４：適切な温度（例えば、８０℃など）で、適切な塩基（例えば、ＮａＯＨなど）の存在下、適切な溶媒（例えば、ＴＨＦ及びＨ_２Ｏなど）中、
工程５：適切な温度（例えば、ＲＴなど）で、適切なアミド縮合試薬（例えば、ＥＤＣＩ及びＨＯＢｔなど）の存在下、適切な塩基（例えば、ＮＭＭなど）の存在下、適切な溶媒（例えば、ＤＣＭなど）中、
工程６：適切な温度（例えば－７０℃など）で、適切な有機金属試薬（例えば、イソプロピルリチウムなど）の存在下、適切な溶媒（例えば、ＴＨＦなど）中、
工程７：適切な温度（例えば９０℃など）で、適切な有機金属触媒（例えば、Ｐｄ（ｄｐｐｆ）Ｃｌ_２など）の存在下、適切な塩基（例えば、Ｎａ_２ＣＯ_３など）の存在下、適切な溶媒（例えば、１，４－ジオキサン及びＨ_２Ｏなど）中、
工程８：適切な温度（例えば、０℃～室温など）で、適切なルイス酸（例えば、ＢＢｒ_３など）の存在下、適切な溶媒（例えば、ＤＣＭなど）中、
工程９：適切な温度（例えば、－７８℃～４０℃など、特に０℃～室温）で、適切な塩基（例えば、ＴＥＡ、ＤＢＵ又はＫ_２ＣＯ_３など）の存在下、適切な溶媒（例えば、ＤＣＭ、ＴＨＦ又はＤＭＦなど）中、

In schemes 1a, 1b and 1c the following reaction conditions apply:
Step 1: In the presence of a suitable base (such as TMEDA) and a suitable organometallic reagent (such as isopropylmagnesium bromide) at a suitable temperature (such as -70°C), in a suitable solvent (such as THF);
Step 2: In the presence of a suitable oxidizing reagent (such as DMP) in a suitable solvent (such as DCM) at a suitable temperature (such as 0° C. to room temperature),
Step 3: In the presence of a suitable organometallic reagent (such as isopropylmagnesium bromide) in a suitable solvent (such as THF) at a suitable temperature (such as −20° C. to room temperature),
Step 4: In the presence of a suitable base (such as NaOH) at a suitable temperature (such as 80° C.) in a suitable solvent (such as THF and H ₂ O);
Step 5: In the presence of a suitable amide condensation reagent (such as EDCI and HOBt) in the presence of a suitable base (such as NMM) in a suitable solvent (such as DCM) at a suitable temperature (such as RT);
Step 6: In the presence of a suitable organometallic reagent (such as isopropyllithium) in a suitable solvent (such as THF) at a suitable temperature (such as -70°C),
Step 7: At a suitable temperature (such as 90° C.), in the presence of a suitable organometallic catalyst (such as Pd(dppf)Cl ₂ ), in the presence of a suitable base (such as Na ₂ CO ₃ ), in a suitable solvent (such as 1,4-dioxane and H ₂ O),
Step 8: In the presence of a suitable Lewis acid (such as _BBr3 ) in a suitable solvent (such as DCM) at a suitable temperature (such as 0° C. to room temperature),
Step 9: In the presence of a suitable base (such as TEA, DBU or K ₂ CO ₃ ) in a suitable solvent (such as DCM, THF or DMF) at a suitable temperature (such as −78° C. to 40° C., in particular 0° C. to room temperature),

In schemes 2a and 2b the following reaction conditions apply:
Step 9: See Scheme 1, Step 9.
Step 10: At a suitable temperature (e.g., room temperature), in the presence of a suitable catalyst (e.g., Pd/C), in the presence of a suitable reducing agent (e.g., H ₂ ), optionally in the presence of a suitable base (e.g., TEA), in a suitable solvent (e.g., THF), or at a suitable temperature (e.g., room temperature), in the presence of a suitable catalyst (e.g., Pd(dppf)Cl ₂ ×DCM complex), a suitable reducing agent (e.g., NaBH ₄ ), a suitable base (e.g., TMEDA), in a suitable solvent (e.g., THF).
Step 11: For N-deprotection, in the presence of a suitable acid (such as, for example, TFA) in a suitable solvent (such as, for example, DCM) at a suitable temperature (such as, for example, room temperature); for O-deprotection, in the presence of a suitable acid (such as, for example, 4-methylbenzenesulfonic acid) in a suitable solvent (such as, for example, MeOH) at a suitable temperature (such as, for example, room temperature);
Step 12: At a suitable temperature (e.g., 80° C., etc.), optionally in the presence of a suitable Lewis acid (e.g., ZnCl ₂ , etc.), in the presence of a suitable reducing agent (e.g., NaBH ₃ CN, etc.), in a suitable solvent (e.g., MeOH, etc.);
Step 13: In the presence of a suitable organometallic catalyst (such as Ag(Phen) ₂ OTf) in the presence of a suitable bromination reagent (such as 1,3-dibromo-1,3,5-triazinane-2,4,6-trione) in a suitable solvent (such as DCE) at a suitable temperature (such as room temperature);
Step 14: In the presence of a suitable chlorinating reagent (such as, for example, oxalyl chloride) in the presence of DMF in a suitable solvent (such as, for example, DCM) at a suitable temperature (such as, for example, room temperature).

In Scheme 3, the following reaction conditions apply:
Steps 11-12: See steps 11 to 12 in Scheme 2.
Step 15: In the presence of a suitable base (e.g. _{, Cs2CO3} ) at a suitable temperature (e.g., 80°C), in a suitable solvent (e.g., DMF, etc.);
Step 16: In the presence of a suitable base (such as, for example, ammonia) in a suitable solvent (such as, for example, 1,4-dioxane) at a suitable temperature (such as, for example, 40° C.).

Part B) Schemes 4, 5, 6, 7, 8, 9, 10, 11 and 12

スキーム４では、以下の反応条件が適用される。
工程１：適切な温度（例えば９０℃など）で、適切な有機金属触媒（例えば、Ｐｄ（ｄｐｐｆ）Ｃｌ_２など）の存在下、適切な塩基（例えば、Ｎａ_２ＣＯ_３など）の存在下、適切な溶媒（例えば、１，４－ジオキサン及びＨ_２Ｏなど）中、
工程２：適切な温度（例えば、ＲＴなど）で、適切なアミド縮合試薬（例えば、ＨＡＴＵなど）の存在下、適切な塩基（例えば、ＤＩＥＡなど）の存在下、適切な溶媒（例えば、ＤＣＭなど）中、
工程３：適切な温度（例えば、－７８℃～室温など）で、適切なルイス酸（例えば、ＢＢｒ_３など）の存在下、適切な溶媒（例えば、ＤＣＭなど）中、
工程４：適切な温度（例えば、－７８℃～４０℃など、特に０℃～室温）で、適切な塩基（例えば、ＴＥＡ、ＤＢＵ又はＫ_２ＣＯ_３など）の存在下、適切な溶媒（例えば、ＤＣＭ、ＴＨＦ又はＤＭＦなど）中、
工程５：適切な温度（例えば、室温など）で、適切な塩基（例えば、ＬｉＯＨ×Ｈ_２Ｏなど）の存在下、適切な溶媒（例えば、ＴＨＦ及びＨ_２Ｏなど）中、
工程６：適切な温度（例えば、室温など）で、適切な有機金属触媒（例えば、Ａｇ（Ｐｈｅｎ）_２ＯＴｆなど）の存在下、適切な臭素化試薬（例えば、１，３－ジブロモ－１，３，５－トリアジナン－２，４，６－トリオンなど）の存在下、適切な溶媒（例えば、ＤＣＥなど）中、
工程７：適切な温度（例えば、室温など）で、適切な臭素化試薬（例えば、１，３－ジブロモ－１，３，５－トリアジナン－２，４，６－トリオン）の存在下、溶媒として２，２，２－トリフルオロエタン－１－オールの存在下。

In Scheme 4, the following reaction conditions apply:
Step 1: At a suitable temperature (such as 90° C.), in the presence of a suitable organometallic catalyst (such as Pd(dppf)Cl ₂ ), in the presence of a suitable base (such as Na ₂ CO ₃ ), in a suitable solvent (such as 1,4-dioxane and H ₂ O),
Step 2: In the presence of a suitable amide condensation reagent (such as HATU) in the presence of a suitable base (such as DIEA) in a suitable solvent (such as DCM) at a suitable temperature (such as RT);
Step 3: In the presence of a suitable Lewis acid (such as _BBr3 ) in a suitable solvent (such as DCM) at a suitable temperature (such as −78° C. to room temperature);
Step 4: In the presence of a suitable base (such as TEA, DBU or K ₂ CO ₃ ) in a suitable solvent (such as DCM, THF or DMF) at a suitable temperature (such as −78° C. to 40° C., in particular 0° C. to room temperature),
Step 5: In the presence of a suitable base (such as LiOH×H ₂ O) in a suitable solvent (such as THF and H ₂ O) at a suitable temperature (such as room temperature);
Step 6: In the presence of a suitable organometallic catalyst (such as Ag(Phen) ₂ OTf) in the presence of a suitable bromination reagent (such as 1,3-dibromo-1,3,5-triazinane-2,4,6-trione) in a suitable solvent (such as DCE) at a suitable temperature (such as room temperature);
Step 7: In the presence of a suitable brominating reagent (eg, 1,3-dibromo-1,3,5-triazinane-2,4,6-trione) in the presence of 2,2,2-trifluoroethan-1-ol as solvent at a suitable temperature (eg, room temperature, etc.).

In Scheme 5, the following reaction conditions apply:
Step 8: In the presence of a suitable base (such as TEA, DBU or K ₂ CO ₃ ) in a suitable solvent (such as DCM, THF or DMF) at a suitable temperature (such as -78°C to 40°C, in particular 0°C to room temperature),
Step 9: In the presence of a suitable base (such as TEA, DBU or K ₂ CO ₃ ) in a suitable solvent (such as DCM, THF or DMF) at a suitable temperature (such as −78° C. to 40° C., in particular 0° C. to room temperature),
Step 10: At a suitable temperature (such as room temperature), in the presence of a suitable organometallic catalyst (such as Pd/C) and a suitable base (such as TEA), in a suitable solvent (such as MeOH) under _H2 atmosphere;
Step 11: When PG is Boc, in the presence of a suitable acid (such as, for example, TFA) in a suitable solvent (such as, for example, DCM) at a suitable temperature (such as, for example, room temperature).

In Scheme 6, the following reaction conditions apply:
Step 12: Reductive amination conditions, at a suitable temperature (e.g., room temperature to 80° C., etc.), in the presence or absence of a suitable Lewis acid (e.g., ZnCl ₂ , etc.) or acid (e.g., AcOH, etc.), in the presence of a suitable reducing agent (e.g., NaBH ₃ CN, etc.), in a suitable solvent (e.g., MeOH, etc.);
Step 13: At a suitable temperature (such as 0° C.), in the presence of a suitable electrophile (such as MsCl), in the presence of a suitable base (such as TEA), in a suitable solvent (such as DCM);
Step 14: In the presence of a suitable oxidizing agent (such as DMP) in a suitable solvent (such as DCM) at a suitable temperature (such as 0° C. to room temperature),
Step 15: In the presence of a suitable acid (e.g., HCl) in a suitable solvent (e.g., ACN) at a suitable temperature (e.g., 50° C., etc.);
Step 16: At a suitable temperature (such as room temperature) in the presence or absence of a suitable base (such as TEA) in a suitable solvent (such as THF).

In Scheme 7, the following reaction conditions apply:
Step 11: When PG is Boc, in the presence of a suitable acid (such as, for example, TFA) in a suitable solvent (such as, for example, DCM) at a suitable temperature (such as, for example, room temperature),
Step 12: Reductive amination conditions, at a suitable temperature (e.g., room temperature to 80° C., etc.), in the presence or absence of a suitable Lewis acid (e.g., ZnCl ₂ , etc.) or acid (e.g., AcOH, etc.), in the presence of a suitable reducing agent (e.g., NaBH ₃ CN, etc.), in a suitable solvent (e.g., MeOH, etc.);
Step 17: In the presence of a suitable base (e.g., _DIEA or _Cs2CO3 ) in a suitable solvent (e.g., DCM or DMF, etc.) at a suitable temperature (e.g., room temperature to 80°C, etc.);
Step 18: In the presence of a suitable base (such as, for example, ammonia) in a suitable solvent (such as, for example, 1,4-dioxane) at a suitable temperature (such as, for example, 40° C.).

In Scheme 8, the following reaction conditions apply:
Step 9: In the presence of a suitable base (such as TEA, DBU or K ₂ CO ₃ ) in a suitable solvent (such as DCM, THF or DMF) at a suitable temperature (such as -78°C to 40°C, in particular 0°C to room temperature),
Step 10: At a suitable temperature (e.g., room temperature, etc.), in the presence of a suitable organometallic catalyst (e.g., Pd/C, etc.), optionally in the presence of a suitable base (e.g., TEA, etc.), in a suitable solvent (e.g., MeOH, etc.) under _H2 atmosphere;
Step 19: In the presence of a suitable chlorinating reagent (such as, for example, oxalyl chloride) in the presence of DMF in a suitable solvent (such as, for example, DCM) at a suitable temperature (such as, for example, room temperature);
Step 20: In the presence of a suitable nucleophilic amine at a suitable temperature (e.g., 90° C., etc.) in a suitable solvent (e.g., EtOH, etc.);
Step 21: In the presence of a suitable acid (such as, for example, HCl in dioxane) in a suitable solvent (such as, for example, MeOH) at a suitable temperature (such as, for example, room temperature);
Step 22: At a suitable temperature (such as 110° C.), in the presence of a suitable boron reagent (such as trimethylboroxine), in the presence of a suitable organometallic catalyst (such as tetrakis(triphenylphosphine)palladium(0)), in the presence of a suitable base (such as K ₂ CO ₃ ), in a suitable solvent (such as 1,4-dioxane),

In Scheme 9, the following reaction conditions apply:
Step 23: In the presence of a suitable base (such as DIEA and n-BuLi) in a suitable solvent (such as THF) at a suitable temperature (such as −78° C. to −25° C.),
Step 24: At a suitable temperature (such as -65°C to -55°C), in the presence of a suitable reducing agent (such as DIBAL-H), in a suitable solvent (such as toluene), preferably in a suitable flow chemistry system;
Step 25: first, at a suitable temperature (such as -10°C to 10°C), in the presence of a suitable base (such as DMAP), in the presence of a suitable condensing agent (such as DCC), in a suitable solvent (such as DCM), then, at a suitable temperature (such as -10°C to 0°C), in the presence of a suitable acid (such as AcOH), in the presence of a suitable reducing agent (such as _NaBH4 ), in a suitable solvent (such as DCM);
Step 26: Heat to reflux in a suitable solvent such as, for example, toluene.
Step 27: In the presence of a suitable reducing agent (such as LiBH ₄ ) in a suitable solvent (such as 2-methyltetrahydrofuran) at a suitable temperature (such as −5° C. to 5° C.),
Step 28: In the presence of a suitable reducing agent (such as NaBH(OAc) ₃ ) in a suitable solvent (such as DCM) at a suitable temperature (such as 15° C. to 25° C.),
Step 29: In the presence of a suitable acid (e.g., HCl) in a suitable solvent (e.g., IPA, etc.) at a suitable temperature (e.g., 15° C. to 25° C., etc.);
Step 30: In the presence of a suitable base (such as, for example, TEA) at a suitable temperature (such as, for example, 5° C. to 30° C.) in the presence of a suitable reducing agent (such as, for example, NaBH(OAc) ₃ ) in a suitable solvent (such as, for example, toluene);
Step 31: In the presence of a suitable base (e.g., K ₂ HPO ₄ , etc.) in a suitable solvent (e.g., H ₂ O, etc.) at a suitable temperature (e.g., 50° C. to 55° C., etc.);
Step 32: When PG is Bn, at a suitable temperature (such as, for example, −5° C. to 45° C.), under a hydrogen atmosphere within a suitable pressure range (such as, for example, 0.27 to 0.40 MPa), in the presence of a suitable catalyst (such as, for example, palladium hydroxide on carbon), in the presence of a suitable acid (such as, for example, MSA), in a suitable solvent (such as, for example, EtOH);
Step 33: In the presence of a suitable base (such as TEA) at a suitable temperature (such as −50° C. to −40° C.), in a suitable solvent (such as 2-methyltetrahydrofuran);
Step 34: In the presence of a suitable base (such as TMG) at a suitable temperature (such as 20° C. to 30° C.), in a suitable solvent (such as 2-methyltetrahydrofuran),
Step 35: under hydrogen atmosphere at a suitable temperature (e.g., 20° C. to 30° C., etc.) and a suitable pressure range (e.g., 0.20 to 0.30 MPa), in the presence of a suitable catalyst (e.g., palladium on carbon, etc.), in a suitable solvent (e.g., MeOH, etc.);
Alternatively, at a suitable temperature (e.g., room temperature), in the presence of a suitable catalyst (e.g., 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex, etc.), a suitable reducing agent (e.g., sodium borohydride), a suitable base (e.g., N,N,N′,N′-tetramethylethylenediamine, etc.), in a suitable solvent (e.g., tetrahydrofuran, etc.).

Scheme 10
In general, compounds of formula (I) where Y ¹ is limited to -CH ₂ - and R ² is limited to W ¹ (hereby designated compounds of formula (Ia)) can be prepared according to the following reaction scheme 10, where W ¹ represents chloro, bromo or iodo, and all other variables are defined according to the scope of the present invention.

In Scheme 10, the following reaction conditions apply:
Step 36: In the presence of a suitable catalyst (e.g. palladium acetate (Pd(OAc) ₂ ) or tris(dibenzylideneacetone)dipalladium(0) (Pd ₂ (dba) ₃ ) or tetrakis(triphenylphosphine)palladium(0)) at a suitable temperature ranging from 60° C. to 100° C. in a suitable solvent (e.g. tetrahydrofuran or dioxane, etc.).

Those skilled in the art will appreciate that starting from compound (Ia), chemistry similar to that reported in step 10 of scheme 5 and steps 20, 21 and 22 of scheme 8 can be carried out.

Scheme 11
In general, compounds of formula (I) where Y ¹ is limited to -CR ^5a R ^5b - and R ² is limited to W ¹ (hereby designated compounds of formula (Ib)) can be prepared according to the following reaction scheme 11, in which at least one of R ^5a and R ^5b is other than hydrogen. All other variables are defined according to the scope of the present invention.

In Scheme 11, the following reaction conditions apply:
Step 37: At a suitable temperature ranging from 80° C. to 200° C., in the presence of a suitable catalyst (e.g., palladium acetate (Pd(OAc) ₂ )), in the presence of a suitable ligand (e.g., triphenylphosphine or tricyclohexylphosphine, etc.), in a suitable solvent (e.g., dioxane, etc.), preferably under sealed conditions, and optionally under microwave irradiation.

Those skilled in the art will appreciate that starting from compound (Ib), chemistry similar to that reported in step 10 of scheme 5 and steps 20, 21 and 22 of scheme 8 can be carried out.

Scheme 12

スキーム１２では、以下の反応条件が適用される。
工程３８：適切な温度（例えば、室温～８０℃など）で、適切な塩基（例えば、ＤＩＥＡ、Ｃ_ｓ２ＣＯ_３、又はＤＢＵ）の存在下、適切な溶媒（例えば、ＤＣＭ、ＴＨＦ又はＤＭＦなど）中、
あるいは、適切な温度（例えば、室温～１００℃など）で、適切な触媒（例えば、Ｐｄ_２ｄｂａ_３など）の存在下、適切な配位子（例えば、キサントホスなど）の存在下、適切な塩基（例えば、Ｃｓ_２ＣＯ_３又はＮａ_２ＣＯ_３）の存在下、適切な溶媒（例えば、ジオキサン又はジオキサンと水の混合物）中。

In Scheme 12, the following reaction conditions apply:
Step 38: In the presence of a suitable base (e.g., DIEA, _Cs2CO3 , or DBU) at a suitable temperature (e.g., room temperature to ₈₀ °C, etc.), in a suitable solvent (e.g., DCM, THF, or DMF, etc.);
Alternatively, at a suitable temperature (e.g., room temperature to 100° C., etc.), in the presence of a suitable catalyst (e.g., Pd ₂ dba ₃ , etc.), in the presence of a suitable ligand (e.g., Xantphos, etc.), in the presence of a suitable base (e.g., Cs ₂ CO ₃ or Na ₂ CO ₃ ), in a suitable solvent (e.g., dioxane or a mixture of dioxane and water).

Those skilled in the art will appreciate that starting from intermediate Z, chemistry similar to that reported when Y ¹ represents O can be carried out.

It will be appreciated that, if appropriate functional groups are present, the compounds of the various formulas, or any intermediates used in their preparation, may be further derivatized by one or more standard synthetic methods employing condensation, substitution, oxidation, reduction, or cleavage reactions. Particular substitution techniques include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation, and coupling procedures.

The compounds of formula (I) may be synthesized in the form of racemic mixtures of enantiomers, which may be separated from one another according to resolution procedures known in the art. Racemic compounds of formula (I) containing basic nitrogen atoms may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. The diastereomeric salt forms are then separated, for example, by selective or fractional crystallization, and the enantiomers are liberated therefrom by alkali. Alternative methods for separating the enantiomeric forms of the compounds of formula (I) include liquid chromatography using chiral stationary phases. The pure stereochemically isomers may also be derived from the corresponding pure stereochemically isomers of the appropriate starting materials, provided that the reaction occurs stereospecifically.

In the preparation of compounds of formula (I), protection of remote functional groups (e.g., primary or secondary amines) of intermediates may be necessary. The need for such protection will vary with the nature of the remote functional group and the conditions of the preparation method. Suitable amino protecting groups (NH-Pg) include acetyl, trifluoroacetyl, t-butoxycarbonyl (Boc), benzyloxycarbonyl (CBz), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection is readily determined by one of ordinary skill in the art. For a general description of protecting groups and their use, see T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 4th ed., Wiley, Hoboken, New Jersey, 2007.

Several methods for preparing compounds of formula (I) are illustrated in the following examples. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification or, alternatively, could be synthesized by one of ordinary skill in the art by using well-known methods.

As will be appreciated by those skilled in the art, compounds synthesized using the protocols shown may exist as solvates, e.g., hydrates, and/or may contain residual solvents or trace impurities. Compounds or intermediates isolated in salt form may be of integer stoichiometry, i.e., mono- or di-salt, or of intermediate stoichiometry. When intermediates or compounds in the experimental part below are shown as "HCl salts" without indicating the number of equivalents of HCl, this means that the number of equivalents of HCl was not confirmed. The same principle can be applied to compounds that are not listed as "oxalates", "formates" or "formates", for example.

」など、実験パートで言及される他の全ての塩形態にも適用される。

" also applies to all other salt forms mentioned in the experimental part.

Those skilled in the art will understand that, even if not explicitly mentioned in the experimental protocols below, typically after column chromatography purification, the desired fractions were collected and the solvent was evaporated.

When stereochemistry is not indicated, this means a mixture of stereoisomers unless otherwise indicated or clear from the context.

When a stereocenter is designated "RS", this means that a racemic mixture was obtained at the indicated center, unless otherwise stated.

Example 1 - Synthesis of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method A
Preparation of intermediate 1-tert-butyl(5-methyl-4-oxohexyl)carbamate

－７０℃に冷却したＴＨＦ（６０ｍＬ）に、ｔｅｒｔ－ブチル２－オキソピロリジン－１－カルボキシラート（５．０ｇ、２７ｍｍｏｌ）及びＴＭＥＤＡ（５．０ｍＬ、３３ｍｍｏｌ）を溶解した溶液に、イソプロピルマグネシウムブロミド溶液（１９ｍＬ、５５ｍｍｏｌ、２－メチルテトラヒドロフラン中２．９Ｍ）をゆっくり添加し、得られた混合物をゆっくりと室温に加温し、１２時間撹拌した。混合物を飽和ＮＨ_４Ｃｌ水溶液（５０ｍＬ）に注ぎ、ＥｔＯＡｃ（５０ｍＬ×３）で抽出した。合わせた有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得、これをＦＣＣ（ＰＥ／ＥｔＯＡｃ＝１：０～１００：１）によって更に精製して、標題中間体（３．７ｇ、収率６０％）を黄色油として得た。

To a solution of tert-butyl 2-oxopyrrolidine-1-carboxylate (5.0 g, 27 mmol) and TMEDA (5.0 mL, 33 mmol) in THF (60 mL) cooled to -70°C, isopropylmagnesium bromide solution (19 mL, 55 mmol, 2.9 M in 2-methyltetrahydrofuran) was slowly added, and the resulting mixture was slowly warmed to room temperature and stirred for 12 h. The mixture was poured into saturated aqueous NH ₄ Cl (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the crude product, which was further purified by FCC (PE/EtOAc = 1:0 to 100:1) to give the title intermediate (3.7 g, 60% yield) as a yellow oil.

Preparation of intermediate 13-tert-butyl 6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate

０℃に冷却したＤＣＭ（１００ｍＬ）に、３，５，６－トリクロロ－１，２，４－トリアジン（１０．０ｇ、５４．２ｍｍｏｌ）及びＴＥＡ（１５．２ｍＬ、１０９ｍｍｏｌ）を溶解した溶液に、ｔｅｒｔ－ブチル２，６－ジアザスピロ［３．４］オクタン－２－カルボキシレート（９．２１ｇ、４３．４ｍｍｏｌ）を添加し、混合物を室温に加温し、１時間撹拌した。混合物を水（２０ｍＬ）で希釈し、ＤＣＭ（３０ｍＬ×３）で抽出した。合わせた有機層をブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得、これをシリカゲルＦＣＣ（ＰＥ／ＥｔＯＡｃ＝１：０～３：１）により精製して、標題中間体（１２．０ｇ、収率５８％）を黄色固体として得た。

To a solution of 3,5,6-trichloro-1,2,4-triazine (10.0 g, 54.2 mmol) and TEA (15.2 mL, 109 mmol) in DCM (100 mL) cooled to 0° C., tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (9.21 g, 43.4 mmol) was added and the mixture was warmed to room temperature and stirred for 1 h. The mixture was diluted with water (20 mL) and extracted with DCM (30 mL×3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product, which was purified by silica gel FCC (PE/EtOAc=1:0-3:1) to give the title intermediate (12.0 g, 58% yield) as a yellow solid.

Preparation of intermediate 27-N-ethyl-5-fluoro-N-isopropyl-2-methoxybenzamide

０℃に冷却した乾燥ＤＣＭ（１５０ｍＬ）に、５－フルオロ－２－メトキシ安息香酸（８．００ｇ、４７．０ｍｍｏｌ）及びＮ－エチルプロパン－２－アミン（８．１９ｇ、９４．０ｍｍｏｌ）を混合した混合物に、ＨＡＴＵ（２１．５ｇ、５６．５ｍｍｏｌ）及びＤＩＥＡ（９．１０ｇ、７０．４ｍｍｏｌ）を小分けにしてゆっくり添加した。得られた混合物をゆっくりと室温に温め、８時間撹拌した。有機層を水（２０ｍＬ×３）で洗浄し、無水Ｎａ_２ＳＯ_４で乾燥させた。濾過後、溶媒を減圧下で除去し、粗生成物をＦＣＣ（ＥｔＯＡｃ／ＰＥ＝０％～２０％）により精製して、標題中間体（１２．０ｇ、収率９６％）を白色固体として得た。

To a mixture of 5-fluoro-2-methoxybenzoic acid (8.00 g, 47.0 mmol) and N-ethylpropan-2-amine (8.19 g, 94.0 mmol) in dry DCM (150 mL) cooled to 0° C., HATU (21.5 g, 56.5 mmol) and DIEA (9.10 g, 70.4 mmol) were slowly added in small portions. The resulting mixture was slowly warmed to room temperature and stirred for 8 h. The organic layer was washed with water (20 mL×3) and dried over anhydrous Na ₂ SO _4. After filtration, the solvent was removed under reduced pressure and the crude product was purified by FCC (EtOAc/PE=0%-20%) to give the title intermediate (12.0 g, 96% yield) as a white solid.

Preparation of intermediate 28-N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide

－７８℃に冷却した乾燥ＤＣＭ（１００ｍＬ）に、Ｎ－エチル－５－フルオロ－Ｎ－イソプロピル－２－メトキシベンズアミド（中間体２７）（１２．０ｇ、５０．１ｍｍｏｌ）を溶解した溶液に、ＢＢｒ_３（１４．４ｍＬ、１５２ｍｍｏｌ）をゆっくり添加し、得られた混合物を室温にゆっくり加温し、８時間撹拌した。混合物を再び－７８℃に冷却し、ＭｅＯＨ（５ｍＬ）を滴加して反応をクエンチした。得られた混合物をゆっくり室温に加温し、飽和ＮａＨＣＯ_３水溶液を添加することによってｐＨ値を約８に調整した。水層をＤＣＭ（５０ｍＬ×３）で抽出し、合わせた有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得て、これをＦＣＣ（ＥｔＯＡｃ／ＰＥ＝０％～２０％）により精製して、標題中間体（９．０ｇ、収率７８％）を白色固体として得た。

To a solution of N-ethyl-5-fluoro-N-isopropyl-2-methoxybenzamide (Intermediate 27) (12.0 g, 50.1 mmol) in dry DCM (100 mL) cooled to −78° C., BBr ₃ (14.4 mL, 152 mmol) was added slowly, and the resulting mixture was slowly warmed to room temperature and stirred for 8 h. The mixture was cooled again to −78° C., and MeOH (5 mL) was added dropwise to quench the reaction. The resulting mixture was slowly warmed to room temperature, and the pH value was adjusted to about 8 by adding saturated aqueous NaHCO ₃ solution. The aqueous layer was extracted with DCM (50 mL×3), and the combined organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the crude product, which was purified by FCC (EtOAc/PE=0%-20%) to give the title intermediate (9.0 g, 78% yield) as a white solid.

Alternative preparation of intermediate 28

ＴＨＦ（１６８Ｌ、１２体積）に５－フルオロ－２－ヒドロキシ－安息香酸（１４．０ｋｇ、８９．６８ｍｏｌ、１．０当量）を混合した混合物を１５～２５℃に調整し、１，１－カルボニルジイミダゾール（１７．４５ｋｇ、１０７．６２ｍｏｌ、１．２当量）を１時間かけて添加した。添加後、混合物を１５～２５℃で１８時間撹拌した。この時間を経た後、Ｎ－エチルプロパン－２－アミン（１４．８５ｋｇ、１７０．３９ｍｏｌ、１．９当量）を混合物に１５～２５℃で２時間かけて添加した。得られた混合物を１５～２５℃で１８～２４時間にわたって更にエージングした。１０％Ｈ_２ＳＯ_４水溶液（１４０ｋｇ、１０体積）を用いてｐＨをｐＨ４～５に調整し、層を分離した。温度を４０℃未満に維持しながら有機相を４２～５６Ｌに濃縮し、次いで、ｎ－ヘプタン（４３ｋｇ、４．５体積）を混合物に１５～２５℃で３時間にわたって添加した。次いで、混合物を０～１０℃に冷却し、更に６時間撹拌した。得られたスラリーを濾過し、ケーキをｔｅｒｔ－ブチルメチルエーテル（ＭＴＢＥ）：ｎ－ヘプタン混合物（２５ｋｇのＭＴＢＥ：ｎ－ヘプタンの２：３体積／体積混合物、２．５体積）で洗浄した。ケーキ洗浄を更に２回繰り返し、得られた固体を真空中５０℃で乾燥させて、中間体２８（１６．５ｋｇ、純度：９９．１％、収率：８０．４％）を得た。

A mixture of 5-fluoro-2-hydroxy-benzoic acid (14.0 kg, 89.68 mol, 1.0 equiv) in THF (168 L, 12 vol) was adjusted to 15-25° C. and 1,1-carbonyldiimidazole (17.45 kg, 107.62 mol, 1.2 equiv) was added over 1 hour. After the addition, the mixture was stirred at 15-25° C. for 18 hours. After this time, N-ethylpropan-2-amine (14.85 kg, 170.39 mol, 1.9 equiv) was added to the mixture over 2 hours at 15-25° C. The resulting mixture was further aged at 15-25° C. for 18-24 hours. The pH was adjusted to pH 4-5 with 10% aqueous H ₂ SO ₄ (140 kg, 10 vol) and the layers were separated. The organic phase was concentrated to 42-56 L while maintaining the temperature below 40° C., then n-heptane (43 kg, 4.5 vol) was added to the mixture over 3 h at 15-25° C. The mixture was then cooled to 0-10° C. and stirred for an additional 6 h. The resulting slurry was filtered and the cake washed with tert-butyl methyl ether (MTBE):n-heptane mixture (25 kg of a 2:3 vol/vol mixture of MTBE:n-heptane, 2.5 vol). The cake wash was repeated two more times and the resulting solid was dried in vacuum at 50° C. to give intermediate 28 (16.5 kg, purity: 99.1%, yield: 80.4%).

Preparation of intermediate 14-tert-butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate

ＴＨＦ（１２０ｍＬ）に、ｔｅｒｔ－ブチル６－（３，６－ジクロロ－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－カルボキシレート（中間体１３）（１２．０ｇ、３３．３ｍｍｏｌ）、Ｎ－エチル－５－フルオロ－２－ヒドロキシ－Ｎ－イソプロピルベンズアミド（中間体２８）（７．５ｇ、３３．３ｍｍｏｌ）及びＤＢＵ（６．１ｇ、４０．１ｍｍｏｌ）を混合した混合物を２５℃で８時間撹拌した。混合物を水（３０ｍＬ）で希釈し、ＤＣＭ（３０ｍＬ×３）で抽出した。合わせた有機層をブラインで洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得、これをＦＣＣ（ＰＥ／ＥｔＯＡｃ＝１：０～３：１）により精製して、標題中間体（１４．０ｇ、収率７３％）を緑色固体として得た。

A mixture of tert-butyl 6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 13) (12.0 g, 33.3 mmol), N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide (intermediate 28) (7.5 g, 33.3 mmol) and DBU (6.1 g, 40.1 mmol) in THF (120 mL) was stirred at 25° C. for 8 h. The mixture was diluted with water (30 mL) and extracted with DCM (30 mL×3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product, which was purified by FCC (PE/EtOAc=1:0 to 3:1) to give the title intermediate (14.0 g, 73% yield) as a green solid.

Preparation of intermediate 2-tert-butyl 6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluoro-phenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate Synthesis method A of intermediate 2

ＴＨＦ（５００ｍＬ）に、ｔｅｒｔ－ブチル６－（３－クロロ－６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４トリアジン－５－イル）－２、６－ジアザスピロ［３．４］オクタン－２－カルボキシレート（中間体１４）（２０ｇ、３６．４ｍｍｏｌ）、ＮａＢＨ_４（２．４８ｇ、６５．７ｍｍｏｌ）及びＴＭＥＤＡ（８．５４ｇ、７３．５ｍｍｏｌ）を混合した混合物に、Ｐｄ（ｄｐｐｆ）Ｃｌ_２×ＤＣＭ（１．７０ｇ、２．０８ｍｍｏｌ）をＮ_２雰囲気下で添加した。添加後、反応混合物を２５℃で１４時間撹拌した。反応混合物を濾過し、濾液を濃縮し、残留物をシリカゲルＦＣＣ（ＥｔＯＡｃ）により精製して、標題中間体（１５ｇ、純度９３％、収率７４％）を褐色固体として得た。

To a mixture of tert-butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 14) (20 g, 36.4 mmol), _NaBH4 (2.48 g, 65.7 mmol) and TMEDA (8.54 g, 73.5 mmol) in THF (500 mL) was added Pd(dppf) _Cl2 x DCM (1.70 g, 2.08 mmol) under _N2 atmosphere. After addition, the reaction mixture was stirred at 25°C for 14 h. The reaction mixture was filtered, the filtrate was concentrated and the residue was purified by silica gel FCC (EtOAc) to give the title intermediate (15 g, 93% purity, 74% yield) as a brown solid.

Synthesis method B for intermediate 2

ＭｅＯＨ（１００ｍＬ）に、ｔｅｒｔ－ブチル６－（３－クロロ－６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－カルボキシレート（中間体１４）（２２．０ｇ、４０．１ｍｍｏｌ）、ＴＥＡ（１５ｍＬ）を溶解した溶液に、Ｐｄ／Ｃ（湿潤、５．０ｇ、１０％）を添加した。得られた混合物をＨ_２雰囲気下（３０ｐｓｉ）、２５℃で８時間撹拌した。反応混合物をｃｅｌｉｔｅパッドを通して濾過し、濾液を真空中で濃縮して、標題中間体（２５．０ｇ、粗製）を得、これを更に精製することなく次工程で直接使用した。

To a solution of tert-butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 14) (22.0 g, 40.1 mmol), TEA (15 mL) in MeOH (100 mL) was added Pd/C (wet, 5.0 g, 10%). The resulting mixture was stirred under _H2 atmosphere (30 psi) at 25 °C for 8 h. The reaction mixture was filtered through a celite pad and the filtrate was concentrated in vacuo to afford the title intermediate (25.0 g, crude), which was used directly in the next step without further purification.

Preparation of intermediate 3-2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropylbenzamide

ＤＣＭ（５ｍＬ）に、ｔｅｒｔ－ブチル６－（６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－カルボキシレート（中間体２）（３００ｍｇ、０．５８３ｍｍｏｌ）を溶解した溶液に、ＴＦＡ（０．５ｍＬ、６．４ｍｍｏｌ）を添加し、得られた混合物を室温で３時間撹拌した。次いで、１０％ＮａＯＨ（５ｍＬ）溶液を混合物にゆっくり添加してｐＨ値を約１２に調整し、得られた混合物をＤＣＭ（１０ｍＬ×３）で抽出した。合わせた有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、真空中で濃縮して、標題中間体（２２０ｍｇ、収率９０％）を白色固体として得た。

To a solution of tert-butyl 6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 2) (300 mg, 0.583 mmol) in DCM (5 mL) was added TFA (0.5 mL, 6.4 mmol) and the resulting mixture was stirred at room temperature for 3 h. Then 10% NaOH (5 mL) solution was added slowly to the mixture to adjust the pH value to about 12, and the resulting mixture was extracted with DCM (10 mL × 3). The combined organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title intermediate (220 mg, 90% yield) as a white solid.

Preparation of compound 61-tert-butyl(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate

ＭｅＯＨ（１５ｍＬ）に、２－（（５－（２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４－トリアジン－６－イル）オキシ）－Ｎ－エチル－５－フルオロ－Ｎ－イソプロピルベンズアミド（中間体３）（１．０ｇ、２．４ｍｍｏｌ）、ｔｅｒｔ－ブチル（５－メチル－４－オキソヘキシル）カルバメート（中間体１）（８３０ｍｇ、３．６２ｍｍｏｌ）及びＺｎＣｌ_２（６６０ｍｇ、４．８４ｍｍｏｌ）を混合した混合物を８０℃で０．５時間撹拌した。次いで、ＮａＢＨ_３ＣＮ（３１０ｍｇ、４．９３ｍｍｏｌ）を添加し、得られた混合物を８０℃で６時間撹拌した。室温に冷却した後、混合物を減圧下で濃縮して粗生成物を得、これをＷａｔｅｒｓ Ｘｂｒｉｄｇｅ Ｐｒｅｐ ＯＢＤを用いた分取ＨＰＬＣ（カラム：Ｃ１８ １５０×４０ｍｍ１０ｕｍ、溶離液：ＡＣＮ／Ｈ_２Ｏ（０．０５％アンモニア）４５％～７５％ｖ／ｖ）により更に精製して、標題化合物（７００ｍｇ、収率４６％）を無色油状物として得た。

A mixture of 2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropylbenzamide (Intermediate 3) (1.0 g, 2.4 mmol), tert-butyl (5-methyl-4-oxohexyl)carbamate (Intermediate 1) (830 mg, 3.62 mmol) and ZnCl ₂ (660 mg, 4.84 mmol) in MeOH (15 mL) was stirred at 80° C. for 0.5 h. NaBH ₃ CN (310 mg, 4.93 mmol) was then added and the resulting mixture was stirred at 80° C. for 6 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give the crude product, which was further purified by preparative HPLC using a Waters Xbridge Prep OBD (column: C18 150×40 mm 10 um, eluent: ACN/H ₂ O (0.05% ammonia) 45%-75% v/v) to give the title compound (700 mg, 46% yield) as a colorless oil.

Preparation of compounds 62 and 63-tert-butyl (R)-(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl) carbamate and tert-butyl (S)-(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl) carbamate

ｔｅｒｔ－ブチル（４－（６－（６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４－トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－イル）－５－メチルヘキシル）カルバマート（化合物６１）（２００ｍｇ、０．３１９ｍｍｏｌ）を、ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＩＧ（カラム：２５０×３０ｍｍ１０ｕｍ、定組成溶離：ＥｔＯＨ（０．１％の２５％アンモニアを含有する）：超臨界ＣＯ_２、４０％：６０％（ｖ／ｖ））を用いたＳＦＣにより精製して、標題化合物（化合物６２）（８５ｍｇ、収率４２％）及び（化合物６３）（８０ｍｇ、収率４０％）をいずれも淡黄色油状物として得た。

tert-Butyl (4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate (compound 61) (200 mg, 0.319 mmol) was purified by SFC using a DAICEL CHIRALPAK IG (column: 250×30 mm 10 um, isocratic elution: EtOH (containing 0.1% of 25% ammonia):supercritical CO ₂ , 40%:60% (v/v)) to give the title compound (compound 62) (85 mg, 42% yield) and (compound 63) (80 mg, 40% yield), both as pale yellow oils.

Compound 64-(R)-2-((5-(2-(6-amino-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropylbenzamide

ＤＣＭ（４ｍＬ）に、ｔｅｒｔ－ブチル（Ｒ）－（４－（６－（６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－イル）－５－メチルヘキシル）カルバメート（化合物６２）（５５０ｍｇ、０．８７６ｍｍｏｌ）を溶解した溶液に、ＴＦＡ（４ｍＬ）をゆっくり添加し、得られた混合物を２５℃で１時間撹拌した。反応混合物を減圧下で濃縮して残渣を得た。残渣をＤＣＭ（４０ｍＬ）で希釈し、ｐＨ値をＮａＯＨ水溶液（２Ｍ、１６ｍＬ）を用いて約１２に調整した。水層をＤＣＭで抽出した（１００ｍＬ×２）。合わせた有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、真空中で濃縮して、標題化合物（４６０ｍｇ、粗製）を黄色固体として得、これを更に精製することなく次工程で直接使用した。

To a solution of tert-butyl (R)-(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate (compound 62) (550 mg, 0.876 mmol) in DCM (4 mL) was added TFA (4 mL) slowly and the resulting mixture was stirred at 25° C. for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was diluted with DCM (40 mL) and the pH value was adjusted to about 12 using aqueous NaOH (2 M, 16 mL). The aqueous layer was extracted with DCM (100 mL×2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound (460 mg, crude) as a yellow solid, which was used directly in the next step without further purification.

Compound 11-(R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

ＤＭＦ（１ｍＬ）に、（Ｒ）－２－（（５－（２－（６－アミノ－２－メチルヘキサン－３－イル）－２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４トリアジン－６－イル）オキシ）－Ｎ－エチル－５－フルオロ－Ｎ－イソプロピルベンズアミド（化合物６４）（１２０ｍｇ、粗製）、１－ブロモ－２－メトキシエタン（３２ｍｇ、０．２３ｍｍｏｌ）、Ｃｓ_２ＣＯ_３（２２２ｍｇ、０．６８１ｍｍｏｌ）、ＮａＩ（１０２ｍｇ、０．６８０ｍｍｏｌ）を混合した混合物を、マイクロ波照射によって８０℃で１時間撹拌した。室温に冷却した後、混合物をＨ_２Ｏ（１０ｍＬ）で希釈し、ＥｔＯＡｃ（３×１０ｍＬ）で抽出した。合わせた有機層をＨ_２Ｏ（１０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で脱水し、濾過し、減圧下に濃縮して粗生成物を得、それを更に、Ｐｈｅｎｏｍｅｎｅｘ Ｇｅｍｉｎｉ－ＮＸ（カラム：１５０×３０ｍｍ５μｍ、溶離液：ＡＣＮ／Ｈ_２Ｏ（１０ｍＭ ＮＨ_４ＨＣＯ_３）５１％～７１％（ｖ／ｖ））を用いたＨＰＬＣにより精製し、更に、ＤＡＩＣＥＬ ＣＨＩＲＡＬＣＥＬ ＯＤ－Ｈ（カラム：２５０×３０ｍｍ５ｕｍ、溶出液：ＥｔＯＨ中の超臨界ＣＯ_２（０．１％ｖ／ｖアンモニア）２５／２５、ｖ／ｖ）を用いたＳＦＣにより精製して、標題化合物（５．１３ｍｇ、純度９６％）を黄色固体として得た。

A mixture of (R)-2-((5-(2-(6-amino-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropylbenzamide (compound 64) (120 mg, crude), 1-bromo-2-methoxyethane (32 mg, 0.23 mmol), Cs ₂ CO ₃ (222 mg, 0.681 mmol), NaI (102 mg, 0.680 mmol) in DMF (1 mL) was stirred at 80° C. for 1 h by microwave irradiation. After cooling to room temperature, the mixture was diluted with H ₂ O (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with H ₂ O (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product which was further purified by HPLC using a Phenomenex Gemini-NX (column: 150×30 mm 5 μm, eluent: ACN/H ₂ O (10 mM NH ₄ HCO ₃ ) 51%-71% (v/v)) and further purified by SFC using a DAICEL CHIRALCEL OD-H (column: 250×30 mm 5 um, eluent: supercritical CO ₂ in EtOH (0.1% v/v ammonia) 25/25, v/v) to give the title compound (5.13 mg, 96% purity) as a yellow solid.

LC-MS (ESI) (Method 1): R _t =2.997 min, m/z observed 586.3 [M+H] ⁺ .

Compound A-(R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

無水ＭｅＯＨ（２ｍＬ）に、（Ｒ－Ｎ－エチル－５－フルオロ－Ｎ－イソプロピル－２－（（５－（２－（６－（（２－メトキシエチル）アミノ）－２－メチルヘキサン－３－イル）－２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４－トリアジン－６－イル）オキシ）ベンズアミド（化合物１１）（４０．０ｍｇ、０．０６８ｍｍｏｌ）、ホルムアルデヒド（５５．４ｍｇ、０．６８３ｍｏｌ、水中３７％）及びＡｃＯＨ（８．２ｍｇ、０．１３７ｍｍｏｌ）を混合した混合物を、４５℃で１時間撹拌した。次いで、ＮａＢＨ_３ＣＮ（８．６ｍｇ、０．１３７ｍｍｏｌ）を混合物に添加し、得られた混合物を４５℃で更に１時間撹拌した。室温に冷却した後、反応混合物を飽和ＮａＨＣＯ_３水溶液（４０ｍＬ）で希釈して、ｐＨ値を約８に調整し、更にＤＣＭ（２０ｍＬ×３）で抽出した。合わせた有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得、これをＢｏｓｔｏｎ Ｐｒｉｍｅ（カラム：Ｃ１８ １５０×３０ｍｍ ５ｕｍ、移動相Ａ：Ｈ_２Ｏ（０．０４％アンモニア＋１０ｍＭ ＮＨ_４ＨＣＯ_３）、移動相Ｂ：ＡＣＮ、流速：２５ｍＬ／分、５０％～８０％（５０％Ｂ～８０％Ｂ）の勾配条件Ｂ／Ａ）を用いた分取ＨＰＬＣにより精製して、標題化合物（９．６２ｍｇ、純度９９．１０％、収率２３．３％）を黄色油状物として得た。

A mixture of (R-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (compound 11) (40.0 mg, 0.068 mmol), formaldehyde (55.4 mg, 0.683 mol, 37% in water) and AcOH (8.2 mg, 0.137 mmol) in anhydrous MeOH (2 mL) was stirred at 45° C. for 1 h. NaBH ₃ CN (8.6 mg, 0.137 mmol) was then added to the mixture and the resulting mixture was stirred at 45° C. for an additional 1 h. After cooling to room temperature, the reaction mixture was diluted with saturated NaHCO The mixture was diluted with ₃ aqueous solution (40 mL) to adjust the pH value to about 8, and further extracted with DCM (20 mL x 3). The combined organic layers were dried over _{anhydrous Na2SO4} , filtered, and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC using Boston Prime (column: C18 150 x 30 mm 5 um, mobile phase A: _H2O (0.04% ammonia + ₁₀ mM _NH4HCO3 ), mobile phase B: ACN, flow rate: 25 mL/min, gradient condition B/A from 50% to 80% (50%B to 80%B)) to give the title compound (9.62 mg, purity 99.10%, yield 23.3%) as a yellow oil.

Example 2 - Synthesis of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method B
Preparation of Intermediate 7-4-((tert-butoxycarbonyl)(methyl)amino)butanoic Acid

ＭｅＯＨ（３０ｍＬ）に、４－（メチルアミノ）ブタン酸塩酸塩（３．０ｇ、１９．５ｍｍｏｌ）及びＴＥＡ（７．７８ｍＬ、５８．６ｍｍｏｌ）を溶解した溶液に、Ｂｏｃ_２Ｏ（４．６９ｇ、２１．５ｍｍｏｌ）を滴下した。混合物を室温で２時間撹拌した。混合物を減圧下で濃縮し、残渣をＥｔＯＡｃ（１００ｍＬ）で希釈し、冷却した０．１Ｎ ＨＣｌ（７０ｍＬ×２）、Ｈ_２Ｏ（５０ｍＬ×２）及びブライン（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、濃縮して、標題中間体（１．８０ｇ、粗製）を無色油状物として得た。

To a solution of 4-(methylamino)butanoic acid hydrochloride (3.0 g, 19.5 mmol) and TEA (7.78 mL, 58.6 mmol) in MeOH (30 mL) was added Boc ₂ O (4.69 g, 21.5 mmol) dropwise. The mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and the residue was diluted with EtOAc (100 mL), washed with cold 0.1 N HCl (70 mL×2), H ₂ O (50 mL×2) and brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated to give the title intermediate (1.80 g, crude) as a colorless oil.

Preparation of intermediate 8-tert-butyl(4-(methoxy(methyl)amino)-4-oxobutyl)(methyl)carbamate

ＣＨＣｌ_３（３０ｍＬ）に、４－（（ｔｅｒｔ－ブトキシカルボニル）（メチル）アミノ）ブタン酸（中間体７）（１．８０ｇ、粗製）を溶解した溶液に、Ｎ，Ｏ－ジメチルヒドロキシルアミン塩酸塩（９６０ｍｇ、９．８４ｍｍｏｌ）、ＨＯＢｔ（１．２４ｇ、９．１８ｍｍｏｌ）及びＮＭＭ（２．８０ｍＬ、２５．１ｍｍｏｌ）を添加した。次いで、ＥＤＣＩ（２．２３ｇ、１１．６ｍｍｏｌ）を添加し、反応混合物を室温で４時間撹拌した。反応混合物をＤＣＭ（１００ｍＬ）で希釈し、１Ｎ ＨＣｌ（３０ｍＬ×３）、飽和ＮａＨＣＯ_３水溶液（３０ｍＬ×３）及びブライン（３０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、真空下で濃縮して、標題中間体（１．７０ｇ、粗製）を無色油状物として得た。

To a solution of 4-((tert-butoxycarbonyl)(methyl)amino)butanoic acid (intermediate 7) (1.80 g, crude) in CHCl ₃ (30 mL) was added N,O-dimethylhydroxylamine hydrochloride (960 mg, 9.84 mmol), HOBt (1.24 g, 9.18 mmol) and NMM (2.80 mL, 25.1 mmol). Then EDCI (2.23 g, 11.6 mmol) was added and the reaction mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM (100 mL), washed with 1N HCl (30 mL×3), saturated aqueous NaHCO ₃ (30 mL×3) and brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title intermediate (1.70 g, crude) as a colorless oil.

Preparation of intermediate 9-tert-butylmethyl(5-methyl-4-oxohexyl)carbamate

Ｎ_２雰囲気下、－７０℃に冷却したＴＨＦ（５ｍＬ）に、ｔｅｒｔ－ブチル（４－（メトキシ（メチル）アミノ）－４－オキソブチル）（メチル）カルバメート（中間体８）（２００ｍｇ、粗製）を溶解した溶液に、イソプロピルリチウム（ペンタン中３．２ｍＬ、２．２４ｍｍｏｌ、０．７Ｍ）を滴下した。得られた混合物を－７０℃で２時間撹拌した。混合物を飽和ＮＨ_４Ｃｌ水溶液（１５ｍＬ）でクエンチし、ＥｔＯＡｃ（３０ｍＬ×２）で抽出した。合わせた有機層をブライン（３０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得た。粗生成物をＦＣＣ（ＰＥ／ＥｔＯＡｃ＝１０：１）により更に精製して、標題中間体（６０ｍｇ）を無色油状物として得た。

To a solution of tert-butyl (4-(methoxy(methyl)amino)-4-oxobutyl)(methyl)carbamate (intermediate 8) (200 mg, crude) in THF (5 mL) cooled to −70° C. under N ₂ atmosphere was added isopropyllithium (3.2 mL, 2.24 mmol, 0.7 M in pentane) dropwise. The resulting mixture was stirred at −70° C. for 2 h. The mixture was quenched with saturated aqueous NH ₄ Cl (15 mL) and extracted with EtOAc (30 mL×2). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product. The crude product was further purified by FCC (PE/EtOAc=10:1) to give the title intermediate (60 mg) as a colorless oil.

Preparation of compound 60-tert-butyl(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)(methyl)carbamate

ＭｅＯＨ（５０ｍＬ）に、２－（（５－（２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４トリアジン－６－イル）オキシ）－Ｎ－エチル－５－フルオロ－Ｎ－イソプロピルベンズアミド（中間体３）（６００ｍｇ、１．４５ｍｍｏｌ）及びｔｅｒｔ－ブチルメチル（５－メチル－４－オキソヘキシル）カルバメート（中間体９）（３３０ｍｇ、１．３７ｍｍｏｌ）を溶解した溶液にＺｎＣｌ_２（７８９ｍｇ、５．７９ｍｍｏｌ）を添加した。得られた混合物を８０℃で２時間撹拌した。次いで、ＮａＢＨ_３ＣＮ（７２９ｍｇ、１１．６ｍｍｏｌ）を添加し、反応混合物を８０℃で一晩撹拌した。室温に冷却した後、混合物を減圧下で濃縮して粗残渣を得、これをＤＣＭ（５０ｍＬ）で希釈し、飽和ＮＨ_４Ｃｌ水溶液（５０ｍＬ）でクエンチし、ＤＣＭ（５０ｍＬ×３）で抽出した。合わせた有機層を食塩水（５０ｍＬ）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、濾液を減圧下で濃縮して粗生成物を得、これをＦＣＣ（ＤＣＭ／ＭｅＯＨ＝１０：１）により更に精製して、標題化合物（４００ｍｇ、収率４２％）が白色固体として得た。

To a solution of 2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropylbenzamide (Intermediate 3) (600 mg, 1.45 mmol) and tert-butyl methyl(5-methyl-4-oxohexyl)carbamate (Intermediate 9) (330 mg, 1.37 mmol) in MeOH (50 mL) was added ZnCl ₂ (789 mg, 5.79 mmol). The resulting mixture was stirred at 80° C. for 2 h. NaBH ₃ CN (729 mg, 11.6 mmol) was then added and the reaction mixture was stirred at 80° C. overnight. After cooling to room temperature, the mixture was concentrated under reduced pressure to give a crude residue, which was diluted with DCM (50 mL), quenched with saturated aqueous NH ₄ Cl (50 mL), and extracted with DCM (50 mL×3). The combined organic layers were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was further purified by FCC (DCM/MeOH=10:1) to give the title compound (400 mg, 42% yield) as a white solid.

Compound 67-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(2-methyl-6-(methylamino)hexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide hydrochloride

ＤＣＭ（１０ｍＬ）に、ｔｅｒｔ－ブチル（４－（６－（６－（２－（エチル（イソプロピル）カルバモイル）－４－フルオロフェノキシ）－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－イル）－５－メチルヘキシル）（メチル）カルバメート（化合物６０）（１ｇ、１．５６ｍｍｏｌ）を溶解した溶液に、ジオキサン中の４Ｍ ＨＣｌ（５ｍＬ、２０ｍｍｏｌ）を添加し、得られた混合物を室温で１時間撹拌した。反応混合物を真空中で濃縮して、標題化合物（９６０ｍｇ、粗製、ＨＣｌ塩）を得、これを更に精製することなく次工程で直接使用した。

To a solution of tert-butyl (4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)(methyl)carbamate (compound 60) (1 g, 1.56 mmol) in DCM (10 mL) was added 4 M HCl in dioxane (5 mL, 20 mmol) and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo to give the title compound (960 mg, crude, HCl salt), which was used directly in the next step without further purification.

Compound A-(R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

ＤＭＦ（５ｍＬ）に、Ｎ－エチル－５－フルオロ－Ｎ－イソプロピル－２－（（５－（２－（２－メチル－６－（メチルアミノ）ヘキサン－３－イル）－２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４トリアジン－６－イル）オキシ）ベンズアミドヒドロクロリド（化合物６７）（４８０ｍｇ、粗製）、Ｋ_２ＣＯ_３（７００ｍｇ、５．０７ｍｍｏｌ）及びＮａＩ（４００ｍｇ、２．６７ｍｍｏｌ）を混合した混合物に、１－ブロモ－２－メトキシエタン（２３０ｍｇ、１．６５ｍｍｏｌ）を添加した。得られた混合物を５０℃で一晩撹拌した。室温に冷却した後、反応混合物をＨ_２Ｏ（３０ｍＬ）でクエンチし、ＤＣＭ（３０ｍＬ×３）で抽出した。合わせた有機層をブライン（３０ｍＬ×３）で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、濾過し、濃縮して粗残渣を得た。残渣をＦＣＣ（ＤＣＭ／ＭｅＯＨ＝１０：１）によって精製し、Ｎ－エチル－５－フルオロ－Ｎ－イソプロピル－２－（（５－（２－（６－（（２－メトキシエチル）（メチル）アミノ）－２－メチルヘキサン－３－イル）－２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４トリアジン－６－イル）オキシ）ベンズアミド（化合物６８）（２５０ｍｇ、収率４８％）を黄色油状物として得た。

To a mixture of N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(2-methyl-6-(methylamino)hexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide hydrochloride (compound 67) (480 mg, crude), K ₂ CO ₃ (700 mg, 5.07 mmol) and NaI (400 mg, 2.67 mmol) in DMF (5 mL) was added 1-bromo-2-methoxyethane (230 mg, 1.65 mmol). The resulting mixture was stirred at 50° C. overnight. After cooling to room temperature, the reaction mixture was quenched with H ₂ O (30 mL) and extracted with DCM (30 mL×3). The combined organic layers were washed with brine (30 mL×3), dried over Na ₂ SO ₄ , filtered and concentrated to give a crude residue. The residue was purified by FCC (DCM/MeOH=10:1) to give N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide (compound 68) (250 mg, 48% yield) as a yellow oil.

N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide (compound 68) (960 mg combined from several batches obtained by method B) was first purified by SFC using a DAICEL CHIRALPAK IG (column: 250×30 mm 10 um, mobile phase: A: supercritical CO ₂ , B: EtOH (0.1% ammonia), A:B=40:60, 60 mL/min) and further purified by SFC using a Boston Prime (column: 150×30 mm 5 um, mobile phase A: H ₂ O (10 mM NH ₄ HCO ₃ ), mobile phase B: ACN, flow rate: 25 mL/min, gradient conditions B/A 55% to 85%) to give the title compound (270 mg) as a colorless oil.

¹ H NMR (400 MHz, methanol-d ₄ ): δ=8.40 (s, 1H), 7.47-7.32 (m, 1H), 7.30-7.10 (m, 2H), 4.24-4.01 (m, 2H), 3.89-3.60 (m, 3H), 3.48 (br s, 3H), 2.63-2.51 (m, 2H), 2.43-2.32 (m, 2H), 2.29-2.07 (m, 6H), 1.86-1.72 (m, 1H), 1.62-1.44 (m, 2H), 1.39-1.02 (m, 10H), 0.99-0.66 (m, 9H). Some protons are hidden by the solvent peaks and are not recorded.

LCMS (ESI) (Method 2): _Rt = 1.965 min, m/z observed 600.3 [M+H] ^<+> .

SFC (Method 11): _Rt = 4.904 min.

Example 3 - Synthesis of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method C
Preparation of Intermediate 22 7-tert-butyl (R)-(1-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-3-methylbutan-2-yl)carbamate

－１０～０℃で予備冷却したＤＣＭ（６０７ｋｇ）中のＢｏｃ－Ｌ－バリン（４４．９ｋｇ）、２，２－ジメチル－１，３－ジオキサン－４，６－ジオン（３２．９ｋｇ）及びＤＭＡＰ（３５．５ｋｇ）を、ＤＣＭ（６１３ｋｇ）にＤＣＣ（５５．５ｋｇ）を溶解した溶液に３時間にわたって添加し、－１０～０℃で１６時間エージンした。温度を１０℃未満に維持しながら、１０％クエン酸水溶液（４４９ｋｇ）を添加した。得られたスラリーを０から１０℃で２時間エージングし、次いで濾過した。濾過ケーキをＤＣＭ（９１ｋｇ）で洗浄した。濾液を分離し、有機層を１０％クエン酸水溶液（４５０ｋｇで２回）及び１０％ＮａＣｌ水溶液（４４９ｋｇ）で洗浄した。有機相（１２００ｋｇ）に、温度を－１０～０℃に維持しながら酢酸（７５．０ｋｇ）を添加した。水素化ホウ素ナトリウム（１８．０ｋｇ）を、温度を－１０℃～０℃の範囲に維持しながら５時間にわたって小分けにして添加し、次いで、得られた混合物を－１０～０℃で更に１６時間エージングした。混合物を１５～２５℃に加温し、２時間エージングした。次いで、混合物を１４％ＮａＣｌ水溶液（４５０ｋｇ）で洗浄し、続いて１４％ＮａＣｌ水溶液（４３２ｋｇ）で２回目の洗浄を行い、最後に水で洗浄した（４４４ｋｇ）。有機相を減圧下で２～４体積まで濃縮した。イソ－プロパノール（１４３ｋｇ）を残渣に添加し、減圧下で４～５体積まで濃縮した。－１０～０℃に冷却し、８時間エージングした後、得られたスラリーを濾過し、ＩＰＡ（３８ｋｇ）で洗浄し、乾燥させて、標題中間体（４６．７ｋｇ、収率６９％）を白色固体として得た。

Boc-L-valine (44.9 kg), 2,2-dimethyl-1,3-dioxane-4,6-dione (32.9 kg) and DMAP (35.5 kg) in DCM (607 kg) pre-cooled at -10 to 0°C were added to a solution of DCC (55.5 kg) in DCM (613 kg) over 3 hours and aged at -10 to 0°C for 16 hours. 10% aqueous citric acid (449 kg) was added while maintaining the temperature below 10°C. The resulting slurry was aged at 0 to 10°C for 2 hours and then filtered. The filter cake was washed with DCM (91 kg). The filtrate was separated and the organic layer was washed with 10% aqueous citric acid (2 x 450 kg) and 10% aqueous NaCl (449 kg). To the organic phase (1200 kg) was added acetic acid (75.0 kg) while maintaining the temperature at -10 to 0°C. Sodium borohydride (18.0 kg) was added in portions over 5 hours while maintaining the temperature in the range of -10 to 0°C, and the resulting mixture was then aged at -10 to 0°C for an additional 16 hours. The mixture was warmed to 15 to 25°C and aged for 2 hours. The mixture was then washed with 14% aqueous NaCl (450 kg), followed by a second wash with 14% aqueous NaCl (432 kg) and a final wash with water (444 kg). The organic phase was concentrated under reduced pressure to 2-4 volumes. Iso-propanol (143 kg) was added to the residue and concentrated under reduced pressure to 4-5 volumes. After cooling to −10 to 0° C. and aging for 8 h, the resulting slurry was filtered, washed with IPA (38 kg) and dried to give the title intermediate (46.7 kg, 69% yield) as a white solid.

Preparation of intermediate 228-tert-butyl (R)-2-isopropyl-5-oxopyrrolidine-1-carboxylate

トルエン（３３３ｋｇ）中のｔｅｒｔ－ブチル（Ｒ）－（１－（２，２－ジメチル－４，６－ジオキソ－１，３－ジオキサン－５－イル）－３－メチルブタン－２－イル）カルバメート（中間体２２７）（４６．７ｋｇ）を加熱還流し、４時間エージングした。混合物を周囲温度に冷却し、濾過し、トルエン（２０ｋｇ）で洗浄した。合わせた濾液を減圧下で濃縮乾固して、所望の化合物（３１．０５ｋｇ、収率９６％）を油状物として得、これを更に精製することなく直接使用した。

tert-Butyl (R)-(1-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-3-methylbutan-2-yl)carbamate (Intermediate 227) (46.7 kg) in toluene (333 kg) was heated to reflux and aged for 4 hours. The mixture was cooled to ambient temperature, filtered, and washed with toluene (20 kg). The combined filtrate was concentrated to dryness under reduced pressure to afford the desired compound (31.05 kg, 96% yield) as an oil which was used directly without further purification.

Preparation of intermediate 229-tert-butyl (5R)-2-hydroxy-5-isopropylpyrrolidine-1-carboxylate

２－ＭｅＴＨＦ（２６．７ｋｇ）中のｔｅｒｔ－ブチル（Ｒ）－２－イソプロピル－５－オキソピロリジン－１－カルボキシレート（中間体２２８）（３０．９ｋｇ）を－５～５℃に冷却した。ＬｉＢＨ_４の２－ＭｅＴＨＦ溶液（１Ｍ，４５．２ｋｇ，５４．４ｍｏｌ）を３時間にわたって添加し、混合物を４時間エージングした。５％ＮａＨＣＯ_３の冷水溶液（１６３ｋｇ）を－５～５℃で３時間にわたって添加し、更に２時間エージングした。混合物を周囲温度に加温し、更に２時間エージングした。水層を分離し、有機層を１０％ＮａＣｌ水溶液（１７０ｋｇ）及び水（１５５ｋｇ）で洗浄した。水洗浄中に、エマルジョンが形成され、分離させるために固体ＮａＣｌ（３．１ｋｇ）を添加した。水層を除去した後、有機層を減圧下で濃縮乾固して、所望の化合物（２８．５ｋｇ、収率９１％）を油状物として得、これを更に精製することなく直接使用した。

tert-Butyl (R)-2-isopropyl-5-oxopyrrolidine-1-carboxylate (Intermediate 228) (30.9 kg) in 2-MeTHF (26.7 kg) was cooled to −5 to 5° C. A solution of LiBH ₄ in 2-MeTHF (1 M, 45.2 kg, 54.4 mol) was added over 3 hours and the mixture was aged for 4 hours. A cold aqueous solution of 5% NaHCO ₃ (163 kg) was added over 3 hours at −5 to 5° C. and aged for an additional 2 hours. The mixture was warmed to ambient temperature and aged for an additional 2 hours. The aqueous layer was separated and the organic layer was washed with 10% aqueous NaCl (170 kg) and water (155 kg). During the water wash, an emulsion formed and solid NaCl (3.1 kg) was added to separate it. After removing the aqueous layer, the organic layer was concentrated to dryness under reduced pressure to give the desired compound (28.5 kg, 91% yield) as an oil, which was used directly without further purification.

Preparation of intermediate 230-tert-butyl (R)-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)carbamate

ＤＣＭ（３４４ｋｇ）中のｔｅｒｔ－ブチル（５Ｒ）－２－ヒドロキシ－５－イソプロピルピロリジン－１－カルボキシレート（中間体２２９）（２８．５５ｋｇ）を１５～２５℃で２－メトキシ－Ｎ－メチルエタン－１－アミン（１２．３ｋｇ、１３８．０ｍｏｌ）で処理し、得られた混合物を１時間エージングした。ナトリウムトリアセトキシボロヒドリド（４０．１２ｋｇ）を、温度を１５～２５℃の間に維持しながら５時間にわたって小分けにして添加し、得られた混合物を４８時間エージングした。温度を１５～２５℃の間に維持しながら、８％ＮａＯＨ水溶液（１８４ｋｇ）を２時間にわたって添加することによって反応混合物をクエンチし、混合物を更に２時間エージングした。水層を分離し、有機層を水（１６９ｋｇ）で洗浄した。次いで、有機層を減圧下で濃縮乾固して、標題中間体（３３．２６ｋｇ、収率８８％）を油状物として得、これを更に精製することなく直接使用した。

tert-Butyl (5R)-2-hydroxy-5-isopropylpyrrolidine-1-carboxylate (Intermediate 229) (28.55 kg) in DCM (344 kg) was treated with 2-methoxy-N-methylethan-1-amine (12.3 kg, 138.0 mol) at 15-25° C. and the resulting mixture was aged for 1 h. Sodium triacetoxyborohydride (40.12 kg) was added in portions over 5 h while maintaining the temperature between 15-25° C. and the resulting mixture was aged for 48 h. The reaction mixture was quenched by the addition of 8% aqueous NaOH (184 kg) over 2 h while maintaining the temperature between 15-25° C. and the mixture was aged for an additional 2 h. The aqueous layer was separated and the organic layer was washed with water (169 kg). The organic layer was then concentrated to dryness under reduced pressure to afford the title intermediate (33.26 kg, 88% yield) as an oil which was used directly without further purification.

Preparation of Intermediate 231-(R)-N ¹ -(2-Methoxyethyl)-N ¹ ,5-dimethylhexane-1,4-diamine, dihydrochloride

イソプロパノール（８４．８０ｋｇ）にＨＣｌを溶解した４モル溶液に、イソプロパノール（２５．６ｋｇ）にｔｅｒｔ－ブチル（Ｒ）－（６－（（２－メトキシエチル）（メチル）アミノ）－２－メチルヘキサン－３－イル）カルバメート（中間体２３０）（３２．３８ｋｇ）を溶解した溶液を周囲温度で３時間にわたって添加し、混合物を周囲温度で更に１９時間エージングした。次いで、メチルｔｅｒｔ－ブチルエーテル（９５．２５ｋｇ）を１時間にわたって添加し、混合物を２．５時間エージングした。得られたスラリーを濾過し、ＭＴＢＥ（５３ｋｇ）で洗浄した。濾過ケーキを乾燥させて、標題化合物（２３．９２ｋｇ、収率８１％）を白色固体として得た。

To a 4 molar solution of HCl in isopropanol (84.80 kg) was added a solution of tert-butyl (R)-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)carbamate (Intermediate 230) (32.38 kg) in isopropanol (25.6 kg) over 3 hours at ambient temperature and the mixture was aged at ambient temperature for an additional 19 hours. Methyl tert-butyl ether (95.25 kg) was then added over 1 hour and the mixture was aged for 2.5 hours. The resulting slurry was filtered and washed with MTBE (53 kg). The filter cake was dried to provide the title compound (23.92 kg, 81% yield) as a white solid.

Preparation of intermediate 23 2-ethyl 1-benzyl-3-(chloromethyl)pyrrolidine-3-carboxylate

－３５～－２５℃に冷却したＴＨＦ（６Ｌ）にＤＩＰＥＡ（９５２ｇ、１．１当量）を溶解した溶液に、－２５℃未満に温度を維持しながらｎ－ＢｕＬｉ（２．３３ｋｇ、ヘキサン中２．５Ｍ、１．０当量）を添加した。得られた混合物を－３５～－２５℃で更に３０分間エージングし、次いで－７８～－６０℃に冷却した。ＴＨＦ（２Ｌ）にエチル１－ベンジルピロリジン－３－カルボキシレート（２ｋｇ、１．０当量）を溶解した溶液を－７８～－６０℃で添加し、更に３０分間撹拌した。次いで、クロロヨードメタン（１．８１ｋｇ、１．２当量）を－７８～－６０℃で投入した。反応混合物を－６０～－４０℃で２時間エージングした。反応混合物を０～１０℃の温度でクエン酸水溶液（６Ｌ Ｈ_２Ｏ中６６０ｇ）に添加し、得られた混合物を２０～３０℃で更に２０分間エージングした。層を分離した後、水層をＥｔＯＡｃ（６Ｌ）で抽出し、合わせた有機層をブライン（６Ｌ）で洗浄し、次いで５０～６０℃に加温した。シュウ酸（２．２２ｋｇ）を５０～６０℃で投入した。得られた混合物を５０～６０℃で３時間撹拌し、次いで、２０～３０℃に冷却し、一晩エージングした。得られた固体を濾過し、ケーキを酢酸エチル（２Ｌ）で洗浄した。ウェットケーキをトルエン（４Ｌ）、Ｈ_２Ｏ（８Ｌ）及びＫ_３ＰＯ_４（１．５当量）に添加し、得られた混合物を２０～３０℃で２０分間エージングした。層を分離した後、水層をトルエン（２Ｌ）で抽出した。有機層を合わせ、水（２Ｌ）で２回洗浄した。有機相を減圧下で濃縮して、４．２ｋｇの所望の化合物をトルエン溶液として得た（アッセイにより４６重量％、アッセイ収率８０％）。

To a solution of DIPEA (952 g, 1.1 equiv.) in THF (6 L) cooled to -35 to -25°C was added n-BuLi (2.33 kg, 2.5 M in hexanes, 1.0 equiv.) while maintaining the temperature below -25°C. The resulting mixture was aged at -35 to -25°C for an additional 30 min and then cooled to -78 to -60°C. A solution of ethyl 1-benzylpyrrolidine-3-carboxylate (2 kg, 1.0 equiv.) in THF (2 L) was added at -78 to -60°C and stirred for an additional 30 min. Chloroiodomethane (1.81 kg, 1.2 equiv.) was then charged at -78 to -60°C. The reaction mixture was aged at -60 to -40°C for 2 h. The reaction mixture was added to aqueous citric acid (660 g in 6 L H ₂ O) at a temperature of 0-10° C., and the resulting mixture was aged at 20-30° C. for another 20 min. After separation of the layers, the aqueous layer was extracted with EtOAc (6 L), the combined organic layers were washed with brine (6 L), and then warmed to 50-60° C. Oxalic acid (2.22 kg) was charged at 50-60° C. The resulting mixture was stirred at 50-60° C. for 3 h, then cooled to 20-30° C. and aged overnight. The resulting solid was filtered and the cake was washed with ethyl acetate (2 L). The wet cake was added to toluene (4 L), H ₂ O (8 L), and K ₃ PO ₄ (1.5 eq.), and the resulting mixture was aged at 20-30° C. for 20 min. After separation of the layers, the aqueous layer was extracted with toluene (2 L). The organic layers were combined and washed twice with water (2 L). The organic phase was concentrated under reduced pressure to give 4.2 kg of the desired compound as a toluene solution (assay 46% by weight, assay yield 80%).

Preparation of intermediate 233-1-benzyl-3-(chloromethyl)pyrrolidine-3-carbaldehyde

流動化学システムで反応を行った。すなわち、トルエン（２６Ｌ）にエチル１－ベンジル－３－（クロロメチル）ピロリジン－３－カルボキシレート（中間体２３２）（４．４ｋｇ）を溶解した溶液を２６．７ｍＬ／分で圧送し、－６０℃に冷却した。冷却後、次いで、これをトルエンにＤＩＢＡＬ－Ｈ（２８．１ｍｏｌ）を溶解した冷却溶液と－６０℃（２８Ｌ）で３２．１ｍＬ／分のポンプ流量で混合した。混合物を－６０℃でパーフルオロアルコキシ（ＰＦＡ）コイル管反応器に通した（合計流量５８．８ｍＬ／分、滞留時間５秒）。得られた混合物を冷却したＭｅＯＨ（－６０℃）と混合し、これを１５．２ｍＬ／分の速度で圧送した。この混合溶液を別のＰＦＡコイルチューブリアクタに－６０℃で圧送した（合計流速７４ｍＬ／分、滞留時間５秒）。得られた混合物を、２０重量％のロッシェル塩水溶液（２０Ｖ）が入ったレシーバーに回収した。層を分離し、有機相を水で２回洗浄した（２×４４Ｌ）。有機相を、同様の方法で調製した別の３．０ｋｇのバッチと合わせ、減圧下で濃縮して、２０．８ｋｇの所望の化合物のトルエン溶液を得（ＨＰＬＣによるアッセイで２５．５重量％、アッセイ収率８５％が得られた）、これを更に精製することなく直接使用した。

The reaction was carried out in a flow chemistry system: a solution of ethyl 1-benzyl-3-(chloromethyl)pyrrolidine-3-carboxylate (Intermediate 232) (4.4 kg) in toluene (26 L) was pumped at 26.7 mL/min and cooled to −60° C. After cooling, it was then mixed with a cooled solution of DIBAL-H (28.1 mol) in toluene at −60° C. (28 L) at a pump rate of 32.1 mL/min. The mixture was passed through a perfluoroalkoxy (PFA) coiled tube reactor at −60° C. (total flow rate 58.8 mL/min, residence time 5 s). The resulting mixture was mixed with chilled MeOH (−60° C.) and pumped at a rate of 15.2 mL/min. This mixed solution was pumped to another PFA coiled tube reactor at −60° C. (total flow rate 74 mL/min, residence time 5 s). The resulting mixture was collected in a receiver containing 20 wt % aqueous Rochelle's salt (20V). The layers were separated and the organic phase was washed twice with water (2 x 44 L). The organic phase was combined with another 3.0 kg batch prepared in a similar manner and concentrated under reduced pressure to give 20.8 kg of a toluene solution of the desired compound (25.5 wt % by HPLC, 85% assay yield), which was used directly without further purification.

^1H NMR (300MHz, Chloroform-d): δ 9.62 (s, 1H), 7.39-7.20 (m, 5H), 3.83-3.57 (m, 4H), 2.96 (d, J=10.2Hz, 1H), 2.80-2.55 (m, 3H), 2.17 (ddd, J=13.9, 7.9, 6.1Hz, 1H), 1.83 (ddd, J=13.4, 7.8, 5.5Hz, 1H).

Preparation of intermediate 234-(R)-4-(6-benzyl-2,6-diazaspiro[3.4]octan-2-yl)-N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine

トルエン（３０Ｌ）及び（Ｒ）－Ｎ^１－（２－メトキシエチル）－Ｎ^１，５－ジメチルヘキサン－１，４－ジアミン、二塩酸塩（中間体２３１）（３．４７ｋｇ）で希釈した、トルエン（３．０ｋｇ、１０重量％）に１－ベンジル－３－（クロロメチル）ピロリジン－３－カルバルデヒド（中間体２３３）を溶解した溶液に、トリエチルアミン（２．５５ｋｇ、２５．２ｍｏｌ）を２０～３０℃で添加した。得られた混合物を２０～３０℃で２時間エージングした。次いで、ナトリウムトリアセトキシボロヒドリド（９．０ｋｇ）を２０～３０℃で投入し、混合物を１２時間エージングした。反応混合物を５～１５℃に冷却し、温度を３５℃未満に維持しながら２５重量％ＮａＯＨ水溶液（２５Ｌ、約１６．７５当量）を添加した。得られた混合物を２０～３０℃で２５分間エージングし、層を分離した。有機層を１５重量％のＮａＣｌ水溶液（１０Ｌ）で洗浄し、層を再度分離し、水（１８Ｌ）を有機相に投入した。水相のｐＨを、４Ｍ ＨＣｌ水溶液を用いて、内部温度を３５℃未満に維持しながら、６～７に調整した。次いで、有機相を廃棄し、水相を分離し、Ｋ_２ＨＰＯ_４でｐＨ８～９に塩基性化した。

To a solution of ¹ -benzyl-3-(chloromethyl)pyrrolidine-3-carbaldehyde (Intermediate 233) in toluene (3.0 kg, 10 wt%) diluted with toluene (30 L) and (R)-N ¹ -(2-methoxyethyl)-N 1 ,5-dimethylhexane-1,4-diamine, dihydrochloride (Intermediate 231) (3.47 kg) was added triethylamine (2.55 kg, 25.2 mol) at 20-30° C. The resulting mixture was aged for 2 hours at 20-30° C. Sodium triacetoxyborohydride (9.0 kg) was then charged at 20-30° C. and the mixture was aged for 12 hours. The reaction mixture was cooled to 5-15° C. and 25 wt % aqueous NaOH (25 L, approximately 16.75 equivalents) was added while maintaining the temperature below 35° C. The resulting mixture was aged at 20-30° C. for 25 min and the layers were separated. The organic layer was washed with 15 wt % aqueous NaCl (10 L), the layers were again separated and water (18 L) was charged to the organic phase. The pH of the aqueous phase was adjusted to 6-7 with 4 M aqueous HCl while maintaining the internal temperature below 35° C. The organic phase was then discarded and the aqueous phase was separated and basified to pH 8-9 with K ₂ HPO ₄ .

The resulting mixture was warmed to 50-55°C and aged for 3 hours. The reaction mixture was then cooled to ambient temperature and combined with the other two batches (2.4 kg + 3.0 kg). The combined stream was washed three times with methyl tert-butyl ether (3 x 40 L). Additional methyl tert-butyl ether (83 L) was added to the resulting aqueous layer and the aqueous phase was basified to pH 9-10 with 8 wt% aqueous NaOH while maintaining a temperature of 15-35°C. The aqueous layer was separated and the organic layer was washed three times with water (3 x 30 L). The organic layer was then concentrated under reduced pressure to approximately 3 volumes, then flushed three times with methanol (3 x 30 L) and concentrated to dryness to give the desired intermediate (12.4 kg, 90% isolated yield) as a pale yellow oil, which was used directly without further purification.

Preparation of intermediate 234a (citrate salt of intermediate 234)

ＥｔＯＨ（８０ｍＬ）及び中間体２３４（２０ｇ）を丸底フラスコに添加した。次に、ＥｔＯＨにクエン酸を溶解した０．５Ｍ溶液（１００ｍＬ、１当量）を丸底フラスコ中の混合物に室温で添加した。その後、混合物を乾燥するまで蒸発させた（ロータリーエバポレータ、４０℃）。アセトニトリル（２００ｍＬ）を残渣に添加し、混合物を乾燥するまで蒸発させた（ロータリーエバポレータ、４０℃）。アセトニトリル（１００ｍＬ）を残渣に添加し、室温で磁気加熱プレートを用いて一晩撹拌した。最後に、中間体２３４ａを濾別し、室温で乾燥させた。

EtOH (80 mL) and intermediate 234 (20 g) were added to a round-bottom flask. Then, a 0.5 M solution of citric acid in EtOH (100 mL, 1 eq.) was added to the mixture in the round-bottom flask at room temperature. The mixture was then evaporated to dryness (rotary evaporator, 40° C.). Acetonitrile (200 mL) was added to the residue and the mixture was evaporated to dryness (rotary evaporator, 40° C.). Acetonitrile (100 mL) was added to the residue and stirred overnight at room temperature using a magnetic heating plate. Finally, intermediate 234a was filtered off and dried at room temperature.

Preparation of the crystalline form of the citrate salt of intermediate 234 (intermediate 234b)

中間体２３４ａ（３．７２ｇ）を室温でアセトニトリル（２０ｍＬ）に添加し、混合物を撹拌した。反応混合物が均一になるまで、混合物を６０℃に加熱した（約１０分）。次に、混合物を０．５℃／分の速度で５０℃まで冷却した。次に、シードを添加し（１９ｍｇの中間体２３４ａ、０．５ｗ／ｗ％）、混合物を撹拌しながら３時間３０分間エージングした。次に、混合物を８時間かけて２０℃まで指数２、３で非線形に冷却した。得られた混合物を一晩撹拌し、生成物を濾別し、乾燥させた（フード内で室温で一晩）。単離後、中間体２３４ｂ（２．７５ｇ；収率７３．９％）を中間体２３４のクエン酸塩の結晶形態として得た。得られた中間体／クエン酸の比は３／２（ＮＭＲ）である。

Intermediate 234a (3.72 g) was added to acetonitrile (20 mL) at room temperature and the mixture was stirred. The mixture was heated to 60° C. until the reaction mixture was homogeneous (approximately 10 min). The mixture was then cooled to 50° C. at a rate of 0.5° C./min. Seeds were then added (19 mg of intermediate 234a, 0.5 w/w%) and the mixture was aged for 3 h 30 min with stirring. The mixture was then non-linearly cooled to 20° C. over 8 h with an exponential of 2:3. The resulting mixture was stirred overnight and the product was filtered off and dried (in a hood at room temperature overnight). After isolation, intermediate 234b (2.75 g; 73.9% yield) was obtained as a crystalline form of the citrate salt of intermediate 234. The resulting intermediate/citric acid ratio is 3/2 (NMR).

The above nonlinear cooling was performed according to the following equation:
A new linear ramp is started every 30 seconds during the prescribed cool down period. The ramp is calculated according to the following formula:

Ｔ_ｓｅｔ：各新規ランプの設定値
Ｔ_{ｓｔａｒｔ ｖａｌｕｅ}：冷却軌跡開始時の混合物温度の測定値
Ｔ_{ｅｎｄ ｖａｌｕｅ}：冷却軌跡の終了規定値
ｔ_{ａｃｔｉｏｎ}：冷却開始からの実時間
Ｄｕｒａｔｉｏｎ：定義された冷却持続時間
ｎ：指数

_Tset : set value for each new ramp _{. Tstartvalue} : measured mixture temperature at the start of the cooling trajectory. _Tendvalue : end _value of the cooling trajectory. taction: actual time since the start of the cooling. Duration: defined duration of the cooling. n: exponent.

^1H NMR (400MHz, MeOH- _d4 ) δ ppm 0.91 (3H, d, J = 6.88Hz) 0.98 (3H, d, J = 6.88Hz) 1.46-1.57 (2H, m) 1.67-1.87 (2H, m) 1.94-2.03 (1H, m) 2.20-2.29 (2H, m) 2.62-2.69 (2H, m) 2.72-2.77 (4H, m) 2.77-2.82 (2H, m) 2.90 (2H, t, J = 7.32Hz) 2.95-3.02 (2H, m) 3.07-3.16 (2H, m) 3.16-3.22 (2H, m) H,m) 3.37 (3 H,s) 3.68-3.72 (2 H,m) 3.83-3.89 (2 H,m) 3.90-3.92 (2 H,m) 3.94-4.06 (2 H,m) 7.32-7.43 (5 H,m).

Preparation of intermediate 224-(R)-N-(2-methoxyethyl)-N,5-dimethyl-4-(2,6-diazaspiro[3.4]octan-2-yl)hexan-1-amine

－５～５℃に冷却したＥｔＯＨ（１．４７ｋｇ）中の水酸化パラジウム炭素（１．２ｋｇ）に、メタンスルホン酸（ＭＳＡ）（１１ｋｇ）、（Ｒ）－４－（６－ベンジル－２，６－ジアザスピロ［３．４］オクタン－２－イル－Ｎ－（２－メトキシエチル）－Ｎ，５－ジメチルヘキサン－１－アミン（中間体２３４）（１０ｋｇ）及びＥｔＯＨ（２５０Ｌ）を添加した。混合物を３５～４５℃に加温し、水素雰囲気（０．２７～０．４０ＭＰａ）下で１６～２０時間撹拌した。混合物を珪藻土（２０ｋｇ）で濾過し、パッドをＥｔＯＨ（２４Ｌ）で洗浄した。濾液を減圧下（４０℃未満）で２～３体積まで濃縮し、次いで、２－ＭｅＴＨＦ（７３ｋｇ及び４７ｋｇ）で２回フラッシングして、２～３体積の溶液を得た。２－ＭｅＴＨＦ（６５ｋｇ）で希釈した後、１０％硫酸ナトリウム水溶液（３０ｋｇ）を添加し、混合物を０～１０℃に冷却し、続いて１６％ＮａＯＨ水溶液（５０ｋｇ）を添加してｐＨを１３～１４に調整した。温度を１５～２５℃に調整し、３０～６０分間撹拌した。水層を分離し、２－ＭｅＴＨＦ（４７ｋｇ×２）で２回抽出した。合わせた有機層を減圧下（４０℃未満）で３～４体積に濃縮し、２－ＭｅＴＨＦ（９５０ｇ）を添加した。減圧下（４０℃未満）で３～４体積に濃縮した後、得られた溶液を２－ＭｅＴＨＦ（３０ｋｇ）で希釈し、４Ａモレキュラーシーブ（２５ｋｇ）に通して乾燥させ、２－ＭｅＴＨＦ（３０ｋｇ）で洗浄した。最終溶液を濃縮して、７９％補正収率で９０．１％のアッセイ純度を有する油状体として所望の化合物（６．７ｋｇ）が得られた。

To palladium hydroxide on carbon (1.2 kg) in EtOH (1.47 kg) cooled to −5 to 5° C. was added methanesulfonic acid (MSA) (11 kg), (R)-4-(6-benzyl-2,6-diazaspiro[3.4]octan-2-yl-N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine (Intermediate 234) (10 kg) and EtOH (250 L). Mixing was performed. The mass was warmed to 35-45°C and stirred under a hydrogen atmosphere (0.27-0.40 MPa) for 16-20 h. The mixture was filtered through diatomaceous earth (20 kg) and the pad was washed with EtOH (24 L). The filtrate was concentrated under reduced pressure (<40°C) to 2-3 volumes and then flushed twice with 2-MeTHF (73 kg and 47 kg) to give 2-3 volumes of solution. After dilution, 10% aqueous sodium sulfate (30 kg) was added and the mixture was cooled to 0-10° C., followed by the addition of 16% aqueous NaOH (50 kg) to adjust the pH to 13-14. The temperature was adjusted to 15-25° C. and stirred for 30-60 min. The aqueous layer was separated and extracted twice with 2-MeTHF (47 kg×2). The combined organic layers were concentrated to 3-4 volumes under reduced pressure (<40° C.) and 2-MeTHF (950 g) was added. After concentration to 3-4 volumes under reduced pressure (<40° C.), the resulting solution was diluted with 2-MeTHF (30 kg), dried through 4A molecular sieves (25 kg) and washed with 2-MeTHF (30 kg). The final solution was concentrated to give the desired compound (6.7 kg) as an oil with an assay purity of 90.1% in 79% corrected yield.

Preparation of intermediate 225-(R)-4-(6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine

（Ｒ）－Ｎ－（２－メトキシエチル）－Ｎ，５－ジメチル－４－（２，６－ジアザスピロ［３．４］オクタン－２－イル）ヘキサン－１－アミン（中間体２２４）（１００ｇ）に、２－ＭｅＴＨＦ（４３０ｇ）及びＴＥＡ（６８ｇ）を添加し、混合物を－５０～－４０℃に冷却した。２－ＭｅＴＨＦ（１７２ｇ）中の３，５，６－トリクロロ－１，２，４－トリアジン（６２ｇ）を添加し、混合物を１～３時間撹拌した。得られた混合物を－２０～－１０℃に加温し、７％ＮａＨＣＯ_３水溶液を添加し、混合物を２０～３０℃に加温し、３０～６０分間撹拌した。水層を除去し、有機層を１０％Ｎａ_２ＳＯ_４（５００ｇ）で洗浄した。有機層を４Åモレキュラーシーブ（２２０ｇ）に通して乾燥させ、２－ＭｅＴＨＦ（１８０ｇ）で洗浄した。標題中間体を、アッセイ収率９０％で、２－１４．８重量％のＭｅＴＨＦの溶液として得た。

To (R)-N-(2-methoxyethyl)-N,5-dimethyl-4-(2,6-diazaspiro[3.4]octan-2-yl)hexan-1-amine (Intermediate 224) (100 g) was added 2-MeTHF (430 g) and TEA (68 g) and the mixture was cooled to -50 to -40°C. 3,5,6-trichloro-1,2,4-triazine (62 g) in 2-MeTHF (172 g) was added and the mixture was stirred for 1-3 hours. The resulting mixture was warmed to -20 to -10°C and 7% aqueous NaHCO ₃ was added and the mixture was warmed to 20-30°C and stirred for 30-60 minutes. The aqueous layer was removed and the organic layer was washed with 10% Na ₂ SO ₄ (500 g). The organic layer was dried through 4 Å molecular sieves (220 g) and washed with 2-MeTHF (180 g) to give the title intermediate as a 2-14.8 wt % solution in MeTHF in 90% assay yield.

Compound 393-(R)-2-((3-chloro-5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)-N-ethyl-5-fluoro-N-isopropyl-benzamide Synthesis method A of compound 393

無水ＴＨＦ（１５ｍＬ）に、Ｎ－エチル－５－フルオロ－２－ヒドロキシ－Ｎ－イソプロピルベンズアミド（中間体２８）（１．１０ｇ、４．８８ｍｍｏｌ）、（Ｒ）－４－（６－（３，６－ジクロロ－１，２，４トリアジン－５－イル）－２，６－ジアザスピロ［３．４］オクタン－２－イル）－Ｎ－（２－メトキシエチル）－Ｎ，５－ジメチルヘキサン－１－アミン（中間体２２５）（１．７０ｇ、３．８２ｍｍｏｌ）及びＤＢＵ（７５０ｍｇ、４．９３ｍｍｏｌ）を混合した混合物を４０℃で８時間撹拌した。室温に冷却した後、混合物を減圧下で濃縮し、得られた残渣をＤＣＭ（６０ｍＬ）で希釈し、Ｈ_２Ｏ（２０ｍＬ×３）で洗浄した。×有機層を無水Ｎａ_２ＳＯ_４で乾燥させ、濾過し、減圧下で濃縮して粗生成物を得、これを精製ＦＣＣ（ＭｅＯＨ／ＤＣＭ＝０％～１０％）により精製して黄色油状物（１．４０ｇ）を得、これを更に、ＤＡＩＣＥＬ ＣＨＩＲＡＬＰＡＫ ＡＤ（カラム：２５０×５０ｍｍ、１０μｍ移動相：Ａ：超臨界ＣＯ_２、Ｂ：ＥｔＯＨ（０．１％アンモニア）、７０ｍＬ／分でＡ：Ｂ＝５０：５０；カラム温度：３８℃；ノズル圧力：１００バール；ノズル温度：６０℃；蒸発器温度：２０℃；トリマー温度：２５℃；波長：２２０ｎｍ）を用いるＳＦＣにより分離して、標題化合物（１．０ｇ）を得た。

A mixture of N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide (Intermediate 28) (1.10 g, 4.88 mmol), (R)-4-(6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine (Intermediate 225) (1.70 g, 3.82 mmol) and DBU (750 mg, 4.93 mmol) in anhydrous THF (15 mL) was stirred at 40° C. for 8 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure and the resulting residue was diluted with DCM (60 mL) and washed with H ₂ O (20 mL×3). ×The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the crude product, which was purified by fine FCC (MeOH/DCM=0%-10%) to give a yellow oil (1.40 g), which was further separated by SFC using DAICEL CHIRALPAK AD (column: 250×50 mm, 10 μm mobile phase: A: supercritical CO ₂ , B: EtOH (0.1% ammonia), A:B=50:50 at 70 mL/min; column temperature: 38° C.; nozzle pressure: 100 bar; nozzle temperature: 60° C.; evaporator temperature: 20° C.; trimmer temperature: 25° C.; wavelength: 220 nm) to give the title compound (1.0 g).

Synthesis method A for compound 393

２０～３０℃の２－ＭｅＴＨＦ（４０ｇ）に、（Ｒ）－４－（６－（３，６－ジクロロ－１，２，４トリアジン－５－イル）－２，６－ジアゾスピロ［３．４］オクタン－２－イル）－Ｎ－（２－メトキシエチル）－Ｎ，５－ジメチルヘキサン－１－アミン（中間体２２５）（２－１４．８重量％のＭｅＴＨＦ溶液６７６ｇ、中間体２２５について補正すると１００ｇ）及びＮ－エチル－５－フルオロ－２－ヒドロキシ－Ｎ－イソプロピルベンズアミド（中間体２８）（５０．６ｇ）を溶解した２－ＭｅＴＨＦ溶液に、テトラメチルグアニジン（３１ｇ）を添加し、混合物を４０～４８時間撹拌した。７％ＮａＨＣＯ_３水溶液（５００ｇ）を添加し、混合物を３０～６０分間撹拌した。水層を除去し、有機層を４％ＮａＯＨ水溶液（２×５００ｇ）で２回洗浄し、１０％Ｎａ_２ＳＯ_４水溶液（５００ｇ）で１回洗浄した。有機層を減圧下（４０℃未満）で２．２～３．０体積まで濃縮し、２－ＭｅＴＨＦ及び含水量の両方が１．０％未満になるまでＭｅＯＨ（１×７９０ｇ及び２×３９５ｇ）で３回フラッシングして、６０．１重量％のメタノール溶液として８６％のアッセイ収率で所望の化合物を得た。××

To a solution of (R)-4-(6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazospiro[3.4]octan-2-yl)-N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine (Intermediate 225) (676 g of a 2-14.8 wt % solution in MeTHF, 100 g corrected for Intermediate 225) and N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide (Intermediate 28) (50.6 g) in 2-MeTHF (40 g) at 20-30° C., tetramethylguanidine (31 g) was added and the mixture was stirred for 40-48 hours. 7% aqueous NaHCO ₃ solution (500 g) was added and the mixture was stirred for 30-60 minutes. The aqueous layer was removed and the organic layer was washed twice with 4% aqueous NaOH (2×500 g) and once with 10% aqueous Na ₂ SO ₄ (500 g). The organic layer was concentrated under reduced pressure (<40° C.) to 2.2-3.0 volumes and flushed three times with MeOH (1×790 g and 2×395 g) until both 2-MeTHF and water content were <1.0% to give the desired compound as a 60.1% wt. solution in methanol in 86% assay yield. XX

Compound A-(R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

（Ｒ）－２－（（３－クロロ－５－（２－（６－（（２－メトキシエチル）（メチル）アミノ）－２－メチルヘキサン－３－イル）－２，６－ジアザスピロ［３．４］オクタン－６－イル）－１，２，４トリアジン－６－イル）オキシ－Ｎ－エチル－５－フルオロ－Ｎ－イソプロピルベンズアミド（化合物３９３）（６０．１重量％のＭｅＯＨ溶液１６３．９３ｇ、化合物３９３について補正すると１００ｇ）、パラジウム炭素（１０ｇ）及びＭｅＯＨ（３１６ｇ）のメタノール溶液を、水素雰囲気（０．２０から０．３０ＭＰａ）下、２０～３０℃で１８時間撹拌した。混合物を珪藻土（７５ｇ）で濾過し、ケーキをＭｅＯＨ（１５８ｇ）で洗浄した。濾液を減圧下（４０℃未満）で約３体積に濃縮し、次いで、酢酸イソプロピル（ＩＰＡｃ、８７０ｇ）でフラッシュして約３体積に濃縮した。次いで、混合物をＩＰＡｃ（６９６ｇ）で希釈し、２０％Ｎａ_２ＣＯ_３水溶液を添加した（５００ｇ）。混合物を３０～６０分間撹拌した。水層を除去した。有機層を水（５００ｇ）で洗浄し、次いで、４５℃未満の減圧下で約３体積に濃縮した。４８．１重量％のＩＰＡｃ溶液として、約９０％のアッセイ収率で標題中間体を得た。

(R)-2-((3-chloro-5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy-N-ethyl-5-fluoro-N-isopropylbenzamide (Compound 393) (163.93 g of a 60.1 wt % solution in MeOH, 100 g corrected for Compound 393), palladium on carbon ( A methanolic solution of 10 g of dimethylformamide (DMSO) and MeOH (316 g) was stirred under hydrogen atmosphere (0.20-0.30 MPa) at 20-30° C. for 18 h. The mixture was filtered through diatomaceous earth (75 g) and the cake was washed with MeOH (158 g). The filtrate was concentrated under reduced pressure (<40° C.) to about 3 volumes and then flushed with isopropyl acetate (IPAc, 870 g) to about 3 volumes. The mixture was then diluted with IPAc (696 g) and 20% aqueous Na ₂ CO ₃ was added (500 g). The mixture was stirred for 30-60 min. The aqueous layer was removed. The organic layer was washed with water (500 g) and then concentrated under reduced pressure at <45° C. to about 3 volumes to give the title intermediate in about 90% assay yield as a 48.1 wt % solution in IPAc.

Example 4 - Synthesis of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy) benzamido oxalate (compound A3)

Compound A3
To a solution of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide (compound A) (270 mg, 0.450 mmol) in 20 mL of ACN (20 mL) was added oxalic acid (81.0 mg, 0.900 mmol). After the addition, the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was then concentrated and the residue was redissolved in ACN and deionized water and lyophilized to provide the title compound (350 mg) as a white solid.

^1H NMR (400MHz, Methanol- _d4 ): δ = 8.48 (s, 1H), 7.52-7.11 (m, 3H), 4.54-3.64 (m, 12H), 3.40-3.34 (m, 5H), 3.23-3.13 (m, 2H), 2.90 (s, 3H), 2.54-2.27 (m, 2H), 2.19-2.03 (m, 1H), 1.97-1.77 (m, 2H), 1.75-1.50 (m, 2H), 1.35-0.65 (m, 17H).

^1H NMR (400MHz, DMSO- _d6 ): δ = 8.51 (s, 1H), 7.51-7.29 (m, 3H), 4.29-3.34 (m, 12H), 3.23-2.84 (m, 7H), 2.70 (s, 3H), 2.35-2.09 (m, 2H), 2.05-1.85 (m, 1H), 1.81-1.58 (m, 2H), 1.56-1.33 (m, 2H), 1.18-0.60 (m, 17H).

LCMS (ESI) (Method 2): _Rt = 1.969 min, m/z observed 600.4 [M+H] ^<+> .

Example 5 - Synthesis of compound A1

ＩＰＡｃ（３６０ｇ）に化合物Ａ（ＩＰＡｃ中４８重量％溶液２０７．９０ｇ、活性化合物Ａ１００ｇ）を溶解した溶液に、ＥｔＯＨ（６３ｇ）を２０℃～２５℃で添加した。次いで、溶液をＥｔＯＨ（４９．５ｇ）中の濃ＨＣｌ（３２．９ｇ）で約１５分間にわたって処理した。混合物に結晶性化合物Ａ１のシード（２ｇ、２％シード負荷）を播種し、次いで、１８時間エージングした。ＩＰＡｃ（８７０ｇ）を２０～２５℃で４時間にわたってゆっくり添加し、スラリーを更に１８時間撹拌した。約５℃に冷却した後、生成物を濾過し、ＩＰＡｃ（５２２ｇ）で洗浄し、２０～３０℃で真空下で乾燥させて、弱結晶性化合物Ａ１を白色固体として得た（収率９１．０％、１１５．４ｇ）。（注：反応に使用した少量のシード材料は、小規模の類似の反応プロトコルを介して得た）

To a solution of Compound A (207.90 g of a 48 wt % solution in IPAc, 100 g of active Compound A) in IPAc (360 g) was added EtOH (63 g) at 20° C.-25° C. The solution was then treated with concentrated HCl (32.9 g) in EtOH (49.5 g) for about 15 min. The mixture was seeded with crystalline Compound A1 seeds (2 g, 2% seed loading) and then aged for 18 h. IPAc (870 g) was added slowly over 4 h at 20-25° C. and the slurry was stirred for an additional 18 h. After cooling to about 5° C., the product was filtered, washed with IPAc (522 g), and dried under vacuum at 20-30° C. to give weakly crystalline Compound A1 as a white solid (91.0% yield, 115.4 g). (Note: A small amount of seed material used in the reaction was obtained via a similar reaction protocol on a smaller scale)

Recrystallization: A solution of weakly crystalline Compound A1 (100 g), EtOH (166 g), purified water (21.5 g) and IPAc (178 g) was stirred at 20-30°C for 0.5 to 2 hours to give a clear solution. Additional IPAc (522 g) was added dropwise over 1-2 hours, and the mixture was then seeded with crystalline Compound A1 seeds (2 g, 2% seed loading). The mixture was then aged for 18-20 hours, IPAc (348 g) was added slowly over 12 hours at 20-30°C, and the slurry was stirred for an additional 55-60 hours. The product was filtered, washed with IPAc (158 g), and dried in vacuum at 20-30°C to give Compound A1 as a white solid (85% yield, 85.0 g net).

^1H NMR (DMSO- _d6 , 400MHz): δ = 11.60 (1H, brs), 10.8 (1H, brs), 8.52 (1H, s), 7.36 (3H, m), 3.97-4.20 (7H, m), 3.64-3.71 (4H, m), 3.47 (7H, m), 3.25 (2H, m), 3.05 (3H, m), 2.73 (3H, s), 2.10-2.45 (1H, m), 1.99 (1H, m), 1.78 (2H, m), 1.55 (2H, m), 0.83-1.12 (12H, m), 0.70 (2H, m).

LCMS (Method 7): _Rt = 0.669 min, m/z observed 600.5 [M+H] ^<+> .

Example 6 - Synthesis of crystal form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bisbesylate hydrate (compound A4) (water equivalent not determined)

４３．０６ｇのベンゼンスルホン酸（遊離塩基化合物Ａに対して２当量）をアセトン／水９５／５ｖ／ｖ混合物８４０ｍＬに添加し、溶解した。ＩＰＡｃに化合物Ａ（８０ｇのＡＰＩを含有する）を溶解した溶液１９２．８ｇを添加した。材料が溶解し、透明な溶液が得られた。更にＩＰＡｃ８０ｍＬを添加し、温度を２５℃に調整した。２％のシードを添加し、混合物を２５℃で１時間撹拌する。次いで、２８．８Ｖ（２３１２ｍＬ）のＩＰＡｃを８時間かけて添加した。その後、懸濁液を２５℃で１８時間撹拌した。懸濁液を濾過し、アセトン／水／ＩＰＡｃ ２３．７５／１．７５／７５ ｖ／ｖ／ｖの混合物３２０ｍＬで洗浄した。１２２．９１ｇの結晶形態Ａビスーベシラート水和物（水当量は求めず）を得た。

43.06 g of benzenesulfonic acid (2 equivalents relative to free base Compound A) was added to 840 mL of acetone/water 95/5 v/v mixture and dissolved. 192.8 g of a solution of Compound A (containing 80 g API) in IPAc was added. The material dissolved and a clear solution was obtained. An additional 80 mL of IPAc was added and the temperature was adjusted to 25° C. 2% seeds were added and the mixture was stirred at 25° C. for 1 hour. Then 28.8 V (2312 mL) of IPAc was added over 8 hours. The suspension was then stirred at 25° C. for 18 hours. The suspension was filtered and washed with 320 mL of acetone/water/IPAc 23.75/1.75/75 v/v/v mixture. 122.91 g of crystalline form A bis-besylate hydrate (water equivalent not determined) was obtained.

Those skilled in the art will appreciate that the small amount of initial seed material used in the above reactions can be obtained via similar reaction protocols on a smaller scale, without adding seeds, by waiting for spontaneous nucleation.

The initial seed of the besylate salt was also obtained during salt screening experiments. In these experiments, 100 mg of the free base was weighed into a 2 mL vial, then 200 μL of ethyl acetate or acetone was added to dissolve the free base. One equivalent of counterion (benzenesulfonic acid) was added to the sample, and the sample was stirred at 25 °C for 3 days. The resulting suspension was centrifuged to obtain the initial seed.

An appropriate amount of crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate was dissolved in deuterated DMSO and a 1D ^1H NMR spectrum was recorded.

One-dimensional proton experiments were collected at 300 K on samples in deuterated DMSO using a Bruker AVANCE NEO-600 MHz NMR spectrometer equipped with a Bruker 5 mm PA BBO 600S3 BB-H-D-05 Z-GRD high-resolution probe and running TOPSPIN 4.0 software.

^1H NMR (600MHz, DMSO- _d6 ) δ ppm 0.69 (br s, 2H) 0.82-0.98 (m, 9H) 1.07 (br s, 4H) 1.31-1.46 (m, 1H) 1.51 (br d, J = 2.91Hz, 1H) 1.69 (br d, J = 3.45Hz, 2H) 1.98 (br s, 1H) 2.06-2.45 (m, 2H) 2.77 (br s, 3H) 2.87-3.19 (m, 3H) 3.24 (br s, 1H) 3.31 (s, 6H) 3.64 (br s, 4H) 3.71-4.59 (m, 7H) H) 7.24-7.54 (m, 9 H) 7.61 (br d, J=7.27 Hz, 4 H) 8.45-8.60 (m, 1 H) 9.24 (br s, 1 H) 9.44-9.82 (m, 1 H).

Example 7 - Alternative synthesis of crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate (Compound A4) (water equivalent not determined)
A 95/5 mixture of isopropanol/water (24 mL) was charged to the flask and heated to 40° C. Benzenesulfonic acid (4.31 g, 98%) was added. This was followed by the addition of 19.3 g of a solution of Compound A in IPAc (containing 8 g of Compound A). Another 16 mL of IPAc was added. 2% seeds were added and the mixture was stirred at 40° C. for 1 hour. IPAc (115.2 mL) was then added dropwise over 8 hours. The mixture was then cooled to 0° C. for 15 hours. The suspension was filtered and the wet cake was washed with (IPA/H ₂ O 95/5)/IPAc 1/6 (32 mL). The wet cake was dried at 25° C. for 16 hours to give 11.44 g of crystalline form A bis-besylate hydrate (water equivalent not determined).

Crystal form A
Crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate can be characterized by its X-ray powder diffraction pattern.

X-ray powder diffraction (XRPD) analysis was performed on a PANalytical Empyrean diffractometer equipped with a Cu-Kα X-ray tube using iCore and dCore variable optics for the incident and diffracted beams, respectively. Compounds were loaded into the cavity of a 16 mm sample holder using the backloading technique.

The samples were subjected to XRPD using the following method:
Tube: Cu:K-α (λ=1.541874 Å)
Generator: Voltage: 45 kV; Current: 40 mA
Geometry: Bragg-Brentano
Scanning mode: Continuous scanning Scanning range: 3-35 degrees Step size: 0.0131 degrees Counting time: 30 s
Spinner rotation time: 1 sec. Injection beam path (iCore)
Program divergence slit: automatic Irradiation length: 10 mm
Soller slit: 0.03 rad
Mask 1: 14mm
Mask 2: 6mm
Width: 7.7mm
Diffraction Beam Path (dCore)
Anti-scattering slit: automatic Irradiation length: 10 mm
Soller slit: 0.04 rad
Detector: PIXcel3D-Medipix3 1x1

Those skilled in the art will recognize that diffraction patterns and peak positions are typically substantially independent of the diffractometer used and the particular calibration method employed. Typically, peak positions may differ by no more than about ±0.2° 2θ. The respective intensities (and relative intensities) of particular diffraction peaks may also vary as a function of various factors, including, but not limited to, particle size, orientation, sample purity, etc.

The X-ray powder diffraction pattern includes peaks at 5.4, 7.2, 11.1, 11.9, and 21.7 degrees 2θ±0.2 degrees 2θ. The X-ray powder diffraction pattern may further include at least one peak selected from 13.7, 14.5, 14.7, 15.0, 16.5, 17.8, 19.0, 19.4, and 20.1 degrees 2θ±0.2 degrees 2θ.

Form A may be further characterized by an X-ray powder diffraction pattern having 4, 5, 6, 7, 8, 9 or more peaks selected from those identified in Table 2.

Form A may be further characterized by an X-ray powder diffraction pattern comprising the peaks identified in Table 2, with the relative intensities of the peaks being greater than about 2%, preferably greater than about 5%, more preferably greater than about 10%, more preferably greater than about 15%. However, one of skill in the art will understand that the relative intensities of the peaks may vary between different samples and between different measurements of the same sample.

Form A may be further characterized by an X-ray powder diffraction pattern substantially as shown in FIG. 1.

Table 2 shows the peak list and relative intensities for the XPRD of crystalline form A of (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4 triazin-6-yl)oxy)benzamide bisbesylate hydrate BSA salt (Figure 1).

Compound A4 described herein is disclosed in International Application No. CN2021/100466 (filed June 17, 2021), the entirety of which is incorporated herein by reference for all purposes.

Analytical Methods The analytical information for the compounds listed above or in the tables below was generated by using the analytical methods described below.

NMR Methods: Some NMR experiments were performed at ambient temperature (298.6 K) using a Bruker Avance III 400 spectrometer equipped with a BBO 400 MHz S1 5 mm probehead with an internal deuterium lock and z-gradients operating at 400 MHz for protons and 100 MHz for carbon. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Some NMR experiments were performed at ambient temperature (298.6 K) using a Varian 400-MR spectrometer equipped with an internal deuterium lock and a Varian 400 4NUC PFG probehead with z-gradients, operating at 400 MHz for protons and 100 MHz for carbon. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Some NMR experiments were performed at ambient temperature (298.6 K) using a Varian 400-VNMRS spectrometer equipped with an internal deuterium lock and a Varian 400 ASW PFG probehead with z-gradients, operating at 400 MHz for protons and 100 MHz for carbon. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Some NMR experiments were performed using a Bruker AVANCE III HD 300 spectrometer at ambient temperature (298.6 K) equipped with an internal deuterium lock and a PA BBO 300S1 BBF-H-D-05 Z 5 mm probehead with z-gradients, operating at 300 MHz for protons and 75 MHz for carbon. Chemical shifts (d) are reported in parts per million (ppm). J values are expressed in Hz.

LCMS (Liquid Chromatography/Mass Spectrometry)
General Procedures High performance liquid chromatography (HPLC) measurements were performed using LC pumps, diode-array (DAD) or UV detectors and columns as specified for each method. HPLC details are shown in Table 3 below. Additional detectors were included where necessary (see Tables 3 and 4 below).

The flow from the column was brought to a mass spectrometer (MS) configured with an atmospheric pressure ion source. It is within the knowledge of one skilled in the art to set tuning parameters (e.g., scan range, dwell time, etc.) to obtain ions that allow identification of the nominal monoisotopic molecular weight (MW) of the compound. Data collection was performed with appropriate software.

Compounds are described by their experimental retention times (R _t ) and ions. Unless otherwise specified in the tables of data, the molecular ions reported correspond to [M+H] ⁺ (protonated molecule) and/or [M−H] ⁻ (deprotonated molecule). If a compound could not be directly ionized, the type of adduct is specified (i.e., [M+NH ₄ ] ⁺ , [M+HCOO] ⁻ , etc.). For molecules with multiple isotopic patterns (Br, Cl), the reported values are those obtained for the lowest isotopic mass. All results were obtained with experimental uncertainties typically associated with the method used.

In the following, "SQD" stands for single quadrupole detector, "RT" stands for room temperature, "BEH" stands for cross-linked ethylsiloxane/silica hybrid, "HSS" stands for high strength silica, and "DAD" stands for diode array detector.

流量（ｍＬ／分）；℃で表されるカラム温度（Ｔ）；実行時間（分）

Flow rate (mL/min); column temperature (T) in °C; run time (min)

Analysis SFC
General Procedure of the SFC Method SFC measurements were performed using an analytical supercritical fluid chromatography (SFC) system configured with a binary pump and modifier to deliver carbon dioxide ( _CO2 ), an autosampler, a column oven, a diode array detector equipped with a high pressure flow cell capable of withstanding up to 400 bar. The analytical SFC details are shown in Table 4 below. When configured with a mass spectrometer (MS), the flow from the column was brought to the (MS). It is within the knowledge of a person skilled in the art to set tuning parameters (e.g. scan range, residence time, etc.) to obtain ions that allow identification of the nominal monoisotopic molecular weight (MW) of the compound. Data collection was performed with appropriate software.

流量（ｍＬ／分）；℃で表されるカラム温度（Ｔ）；実行時間（分）、バール単位の背圧（ＢＰＲ）又はポンド重／平方インチ（ｐｓｉ）。「ＡＣＮ」はアセトニトリルを意味し、「ＭｅＯＨ」はメタノールを意味し、「ＥｔＯＨ」は、エタノールを意味し、「ＤＥＡ」はジエチルアミンを意味する。上記の表４で使用される他の全ての略語は、前に定義した通りである。

Flow rate (mL/min); column temperature (T) in °C; run time (min), back pressure in bar (BPR) or pounds force per square inch (psi). "ACN" means acetonitrile, "MeOH" means methanol, "EtOH" means ethanol, and "DEA" means diethylamine. All other abbreviations used in Table 4 above are as previously defined.

Pharmacology Part 1) Menin/MLL Homogeneous Time-Resolved Fluorescence (HTRF) Assay To a non-treated white 384-well microtiter plate was added 40 nL of 200× test compound in DMSO and 4 μL of 2× terbium chelate-labeled menin (preparation see below) in assay buffer (40 mM Tris·HCl, pH 7.5, 50 mM NaCl, 1 mM DTT (dithiothreitol) and 0.05% Pluronic F-127). After incubation of the test compounds and terbium chelate-labeled menin for 30 minutes at ambient temperature, 4 μL of 2× FITC-MBM 1 peptide (FITC-β-alanine-SARWRFPARPGT-NH ₂ ) ("FITC" means fluorescein isothiocyanate) in assay buffer was added, the microtiter plate was centrifuged at 1000 rpm for 1 minute, and the assay mixture was incubated for 15 minutes at ambient temperature. The relative amount of menin FITC-MBM1 complex present in the assay mixture was determined by measuring the homogeneous time-resolved fluorescence (HTRF) of the terbium/FITC donor/acceptor fluorophore pair using an EnVision microplate reader (excitation 337 nm/terbium emission 490 nm/FITC emission 520 nm) at ambient temperature. The extent of fluorescence resonance energy transfer (HTRF value) is expressed as the ratio of the fluorescence emission intensities of FITC and the terbium fluorophore (F ^<em> 520 nm/F ^<em> 490 nm). The final concentrations of reagents in the binding assay are 200 pM terbium chelate-labeled menin, 75 nM FITC-MBM1 peptide, and 0.5% DMSO in assay buffer. Dose-response titrations of test compounds are performed using an 11-point, four-fold serial dilution scheme, typically starting at 10 μM.

The efficacy of the compounds was first determined by calculating the % inhibition at each compound concentration according to Equation 1.
% Inhibition=((HC-LC)−(HTRF ^compound -LC))/(HC-LC)) ^* 100 (Equation 1)
where LC and HC are the HTRF values of the assay in the presence or absence of a saturating concentration of a compound that competes with FITC-MBM1 for binding to menin, and HTRF ^Compound is the HTRF value measured in the presence of the test compound. HC and LC HTRF values represent the average of at least 10 replicates per plate. For each test compound, the % inhibition values were plotted against the logarithm of the test compound concentration, and the _IC50 value obtained from fitting these data to Equation 2 was plotted.
Inhibition %=Bottom+(Top-Bottom)/(1+10^((logIC ₅₀ -log[cmpd]) ^* h)) (Equation 2)
where Bottom and Top are the lower and upper asymptote of the dose-response curve, respectively, IC ₅₀ is the concentration of compound that results in 50% inhibition of the signal, and h is the Hill coefficient.

Preparation of terbium cryptate labeling of menin: Menin (a.a1-610-6xhis tag, 2.3 mg/mL in 20 mM Hepes (2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid), 80 mM NaCl, 5 mM DTT (dithiothreitol), pH 7.5) was labeled with terbium cryptate as follows: 200 μg menin was buffer exchanged into 1× Hepes buffer. 6.67 μM menin was incubated with an 8-fold molar excess of NHS (N-hydroxysuccinimide)-terbium cryptate for 40 min at room temperature. Half of the labeled protein was purified from the free label by running the reaction on a NAP5 column with elution buffer (0.1 M Hepes, pH 7 + 0.1% BSA (bovine serum albumin)). The remaining half was eluted with 0.1 M phosphate-buffered saline (PBS), pH 7. 400 μl of each eluate was collected, aliquoted, and frozen at −80° C. The final concentrations of terbium-labeled menin protein were 115 μg/mL in Hepes buffer and 85 μg/mL in PBS buffer, respectively.

Menin protein sequence (SEQ ID NO:1):
MGLKAAQKTLFPLRSIDDVVRLFAAELGREEDLVLLSLVLGFVEHFLAVNRVIPTNVPELTFQPSPAPDPPPGGLTYFPVADLSIIAALYARFTAQIRGAVDLSLYPREGGVSSRELEVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITG TKLDS SGVAFAVVGACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTVNAGVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLWLLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIAS AKTYYRDEHIYPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVANDVIPNLLKEAASLLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKWEEGSPTPVLHVGWATFLVQSLGRFEGQVRQKVRIVSREA EAAEAEEPWGEEEAREGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPEGGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQVQMKKQKVSTPSDYTLSFLKRQRKGLHHHHH

2a) Proliferation Assays The antiproliferative effects of menin/MLL protein/protein interaction inhibitor test compounds were evaluated in human leukemia cell lines. The cell line MOLM-14 harbors an MLL translocation and expresses the MLL fusion protein MLL-AF9 and the wild-type protein from the second allele, respectively. OCI-AML3 cells, which harbor an NPM1c gene mutation, were also tested. MLL-rearranged cell lines (e.g. MOLM-14) and NPM1c mutant cell lines display an elevated stem cell-like HOXA/MEIS1 gene expression signature. To exclude compounds showing general cytotoxic effects, KO-52 was used as a control cell line containing two MLL (KMT2A) wild-type alleles.

MOLM-14 cells were cultured in RPMI-1640 (Sigma Aldrich) supplemented with 10% heat-inactivated fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg/ml gentamicin (Gibco). KO-52 and OCI-AML3 cell lines were grown in alpha-MEM (Sigma Aldrich) supplemented with 20% heat-inactivated fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich), and 50 μg/ml gentamicin (Gibco). Cells were maintained at 0.3-2.5 million cells/mL in culture and did not exceed 20 passages.

To evaluate the anti-proliferative effect, 200 MOLM-14 cells, 200 OCI-AML3 cells or 300 KO-52 cells were seeded in 200 μl medium/well in 96-well round-bottom ultra-low attachment plates (Costar, cat. no. 7007). The cell seeding number was selected based on the growth curve to ensure linear growth throughout the experiment. Test compounds were added at different concentrations and DMSO content was normalized to 0.3%. Cells were incubated for 8 days at 37°C and 5% _CO2 . Spheroid-like growth was measured in real time by live cell imaging (IncuCyteZOOM, Essenbio, 4x objective) acquiring images on day 8. Confluence (%) as a measure of spheroid size was determined using the integrated analysis tool.

To determine the effect of test compounds over time, the confluence as a measure of spheroid size in each well was calculated. The confluence of the highest dose of reference compound was used as the baseline for LC (low control) and the confluence of DMSO-treated cells was used as 0% cytotoxicity (high control, HC).

Absolute _IC50 values were calculated as the percentage change in confluence as follows:
LC = low control: cells treated with, e.g., 1 μM of the cytotoxic agent staurosporine or, e.g., cells treated with a high concentration of a surrogate reference compound;
HC = high control: average confluence (%) (DMSO-treated cells);
% Effect=100-(100 ^* (Sample-LC)/(HC-LC)); and _IC50 was calculated using GraphPad Prism (Version 7.00). A dose-response equation was used to plot % Effect vs. Log10 compound concentration with a variable slope, with the maximum fixed at 100% and the minimum fixed at 0%.

2b) MEIS1 mRNA Expression Assay MEIS1 mRNA expression upon compound treatment was examined by Quantigene Singleplex assay (Thermo Fisher Scientific). This technology allows direct quantification of mRNA targets using probes hybridizing to a defined target sequence of interest, and the signal is detected using the multimode plate reader Envision (PerkinElmer). MOLM-14 cell line was used for this experiment. Cells were seeded at 3,750 cells/well in 96-well plates in the presence of increasing concentrations of compounds. After 48 hours of incubation with compounds, cells were lysed in lysis buffer and incubated at 55°C for 45 minutes. Cell lysates were mixed with a human MEIS1-specific capture probe or a human RPL28 (ribosomal protein L28)-specific probe as a normalization control, as well as a blocking probe. The cell lysates were then transferred to custom assay hybridization plates (Thermo Fisher Scientific) and incubated at 55°C for 18-22 hours. The plates were then washed to remove unbound material, followed by sequential addition of preamplifier, amplifier and labeled probe. Signals (=gene counts) were measured on a multimode plate reader Envision. _IC50s were calculated by dose-response modeling using appropriate software. For all non-housekeeper genes, the response is equal to the counts corrected for background and relative expression. For each sample, each test gene signal (background subtracted) was divided by the normalized gene signal (RPL28: background subtracted). Fold changes were calculated by dividing the normalized values of treated samples by the normalized values of DMSO-treated samples. The fold changes of each target gene were used to calculate _IC50 .

The results are summarized in Table 5 below.

3) Mouse PK (in vivo T1 _/2 and oral bioavailability)
In vivo pharmacokinetics (PK) was evaluated following single intravenous (IV, 0.5 or 1.0 mg/kg administered intravenously in 2.5 ml/kg) or oral (PO, 5 mg/kg administered in 10 mL solution/kg) administration of test articles formulated in HP-β-CD 20% (w:vol) solution or pyrogen-free water to fasted male CD-1 mice (6-8 weeks old).

Plasma and/or whole blood samples were collected from the dorsal metatarsal vein at the desired time points by continuous capillary microsampling (approximately 0.03 mL) using EDTA as an anticoagulant. Compound concentrations in plasma and blood samples were analyzed using a qualified LC-MS/MS method. In silico analysis of the main pharmacokinetic parameters was performed using WinNonlin (Phoenix™, version 6.1) or similar software.

4) Metabolic Stability in Human/Mouse Liver Microsomes Experimental Procedure The purpose of this experiment was to measure the in vitro metabolic stability of test compounds in human and mouse liver microsomes and to provide quantitative information on the turnover rate (i.e., to determine the apparent intrinsic clearance of the test compounds).

Test items were prepared at a stock concentration of 10 mM in DMSO. To study metabolic turnover, final working solutions were prepared by adding 2 μL of a 10 mM DMSO stock solution for the test compound or positive control compound to 198 μL of acetonitrile (final concentration 100 μM).

Incubations were performed as follows: First, liver microsomes were thawed on ice and a master solution containing liver microsomes in 100 mM PBS (phosphate buffered saline) was prepared at pH 7.4. Then, the liver microsome solution was added to the incubation plate and 10 mM NADPH (nicotinamide adenine dinucleotide phosphate) was added (MW: 833.4 g/mol, Roche Diagnostics GmbH, Germany. Dissolved in phosphate buffer (100 mmol/L, pH 7.4)). The mixture was mixed for 10 seconds and preheated at 37° C. for 10 minutes in the incubation plate. 5 μL of a 100 μM working solution of the test compound or positive control compound was added to the incubation plate to start the metabolic reaction (final test item concentration = 1 μM). The final reaction mixture should contain 1 mM NADPH, 0.5 mg/mL microsomal protein, and 1 μM test compound or positive control compound in 100 mM PBS, pH 7.4. The percentage of organic solvent in the incubation mixture is 1% and DMSO≦0.02%.

Reactions were quenched at selected time points by transferring 50 μL of the incubated mixture to a quench plate containing 200 μL of cold methanol. After all time points were sampled, the quench plate was centrifuged at 4000 rpm for 40 minutes to precipitate proteins. A total of 90 μL of supernatant was transferred to the analysis plate and ultrapure _H2O was added to each well for LC/MS/MS analysis. All incubations and analyses were performed in duplicate.

Data Analysis All calculations were performed using Microsoft Excel. Slope values k were determined by linear regression of the natural logarithm of the percentage parent drug remaining versus incubation time curves. The results are summarized in Table 6 below.

The in vitro half-life (in vitro t _1/2 ) was determined from the slope value.
In vitro t _1/2 =-(0.693/k)
Conversion of in vitro t _1/2 (min) to in vitro intrinsic clearance (in vitro CL _int , in μL/min/mg protein) was performed using the following formula:

「ＮＡ」は分析しなかったことを意味する

"NA" means not analyzed

5) Protocol for Pharmacodynamic (PD) Activity in Subcutaneous (sc or SC) Xenografts of MOLM-14 or OCI-AML3 Cells Test Agents and Controls Compound A3 was formulated in 20% hydroxypropyl-beta-cyclodextrin (HP-β-CD) and prepared to reach a total volume of 0.2 mL per dose (10 mL/kg) for a 20 g animal. Doses were adjusted by individual body weight daily. Working stocks of Compound A3 were prepared once a week for each experiment and stored at room temperature. Compound A3 was administered daily orally (PO).

Assays In vivo pharmacodynamic (PD) activity of compounds was evaluated in subcutaneous (SC) xenografts of MOLM-14 or OCI-AML3 cells. Nude NMRI mice (Crl: NMRI-Foxn1nu/-) bearing MOLM-14 or OCI-AML3 tumors were treated with vehicle or compound three times daily. Plasma samples were collected 23 hours after the second dose, 0.5 hours after the final dose, and 16 hours after the final dose, and tumor samples were collected 16 hours after the final dose on day 2. To examine the effect of compounds on the expression of multiple menin-KMT2A target genes (e.g., MEIS1, MEF2C, FLT3), QuantiGene Plex technology (Thermo Fisher Scientific) was used. Frozen tumors were homogenized and transferred to individual lysis matrix tubes in lysis buffer and incubated at 55°C for 30 minutes. Cell lysates were mixed with target-specific capture probes, Luminex beads and blocking probes, transferred to custom assay hybridization plates (Thermo Fisher Scientific) and incubated at 54°C for 18-22 hours. The plates were then transferred to magnetic separation plates and washed to remove unbound material from the beads, followed by sequential hybridization consisting of preamplifier, amplifier and labeled probe, followed by streptavidin-phycoerythrin binding. Signal from the beads was measured on a Luminex FlexMap three-dimensional instrument. For all non-housekeeper genes, responses equal counts corrected for background and relative expression. For each sample, each test gene signal (background subtracted) was divided by the normalized gene signal (RPL19, RPL28, ATP6V1A, background subtracted). Fold changes were calculated by dividing the normalized values of treated samples by the normalized values of DMSO-treated samples. The results are summarized in Tables 7 and 8 below.

Tables 7a and 8a show median values based on replicate experiments under optimized conditions using fresh tumor samples.

6) Efficacy Studies in MOLM-14 Subcutaneous Model Test Agents and Controls Compound A3 was formulated in 20% hydroxypropyl-beta-cyclodextrin (HP-β-CD) and prepared to reach a total volume of 0.2 mL per dose (10 mL/kg) for a 20 g animal. Doses were adjusted daily by individual body weight. Working stocks of Compound A3 were prepared once a week for each experiment and stored at 25°C.

Animals Female NMRI nude mice (MOLM-14 SC) were used when approximately 6-8 weeks old and weighing approximately 25 g. All animals were allowed to acclimate to and recover from any shipping-related stress for a minimum of 7 days prior to experimental use. Autoclaved water and irradiated food were provided ad libitum and animals were maintained on a 12-hour light/dark cycle. Cages, bedding, and water bottles were autoclaved prior to use and changed weekly. Further details are provided in Table 9 below.

Tumor Models and Cell Culture Methods Human AML MOLM-14 cells were cultured in the indicated complete culture medium (RPMI 1640 + 10% HI-FBS + 2 mM L-glutamine + 50 ug/ml gentamicin) at 37°C, 5% _CO2 . Cells were harvested during logarithmic growth and resuspended in cold (4°C) Roswell Park Memorial Institute in serum-free medium (RPMI) 1640.

Each mouse was administered 5×10 ⁶ MOLM-14 cells in 50% Matrigel in a total volume of 0.2 mL using a 1 cc syringe and 27 gauge needle into the right flank.

Experimental Design Compound A3 was administered orally (PO) daily.

Day 0 is the day of tumor cell implantation and the start of the experiment.

Mice bearing SC MOLM-14 tumors were randomized 16 days after tumor implantation and assigned to treatment groups according to tumor volume (mean ≈130 mm ³ , n=10/group). Treatment with vehicle or Compound A3 (30 and 100 mg/kg) was initiated on the same day and administered orally daily for 21 days. Plasma was collected for PK analysis at 1, 2, 4, 8, and 23 hours after the last dose (n=4-5/group/time point).

Animal Monitoring SC tumor volumes were measured for each animal two to three or more times weekly throughout the experiment.

Calculations Tumor volumes were calculated using the following formula:
Tumor volume ( ^mm3 ) = (D x ^d2 /2), where "D" represents the larger diameter and "d" represents the smaller diameter of the tumor as measured by caliper measurements. Tumor volume data were graphed as mean tumor volume ± SEM.

ΔTGI% was defined as the difference between the mean tumor burden of the treatment and control groups and was calculated _as ΔTGI%=([( _{TVcTVc0 )} ( _{TVtTVt0 )} ]/( _TVcTVc0 ))×100, where "TVc" is the mean tumor burden of a given control group, " _TVc0 " is the mean initial tumor burden of a given control group, "TVt" is the mean tumor burden of the treatment group, and " _TVt0 " is the mean initial tumor burden of the treatment group. TGI% was defined as the difference between the mean tumor burden of the treatment and control groups and was calculated as ΔTGI%=([(TVcTVc0)(TVtTVt0)]/(TVcTVc0))×100, where " _TVc " is the mean tumor burden of a given control group, "_{TVc0" is the mean initial tumor burden of a given control group, "TVt" is the mean tumor burden of the treatment group, and "TVt0} " is the mean initial tumor burden of the treatment group.

The mean tumor volumes for the treatment and control groups were calculated as follows:
TGI%=(( _{TVcTVt )} / _TVc )×100, where " _TVc " is the mean tumor volume of the control group and " _TVt " is the mean tumor volume of the treatment group. As defined by the National Cancer Institute criteria, a TGI of 60% or greater is considered biologically significant.

Tumor regression (TR) %, quantified to reflect treatment-related reduction in tumor volume compared to baseline independent of the control group, was calculated as TR % = (1 - mean (TV _t i/TV _t0 i)) x 100, where "TV _t i" is the tumor burden of individual animals in the treatment group and "TV _t0 i" is the initial tumor burden of the animal.

Data Analysis Tumor volumes were graphed using Prism software (GraphPad version 7 or 8). Statistical significance of most experiments was assessed for Compound A3-treated groups compared to HPβCD vehicle-treated controls on the last day of the experiment when more than ⅔ of mice remained in each group. Differences between groups were considered significant when p≦0.05.

Statistical significance of animal tumor volumes was calculated using linear mixed-effects (LME) analysis in R software version 3.4.2 (using Janssen's internally developed Shiny application version 4.0), with treatment and time as fixed effects and animal as a random effect. If individual longitudinal response trajectories were not linear, logarithmic transformation was performed.

Information obtained from this model was used to perform pairwise comparisons of tumor volume in the control group or between all treatment groups. The results are shown in Figure 2.

7) Electrophysiological Effects of Test Compounds on Synchronized Beating of Human Pluripotent Stem Cell-Derived Cardiomyocytes (hSC-CM) Using Ca ²⁺ Fluorescence Assay (CTCM Human) Protocol Compounds were tested in 96-well plates.

Compounds were tested at 0.1 μM, 0.2 μM, 0.5 μM, 1 μM, 2.5 μM and 5 μM (n=4/dose) for Cor. 4U®-cardiomyocytes or iCell® cardiomyocytes 2.

Alternatively, primarily for iCell® Cardiomyocytes 2, compounds were tested at 0.1 μM, 0.3 μM, 1 μM, 3 μM, 10 μM and 30 μM (n=4/dose).

Positive and negative controls 3 nM dofetilide 100 nM isoproterenol 100-300 nM nimodipine 3 μM cetirizine.

Vehicle control: dimethylsulfoxide (DMSO). Solution of compound in DMSO or its solvent (final concentration of 0.1% DMSO, n=8).

Preparation of Test Substances and Controls Tested compounds were dissolved in DMSO at 1000x the intended concentration. Compound "mother plates" were made containing test compounds as well as positive and negative controls at 1000x the final concentration. On the day of the experiment, these stock solutions were diluted (in round bottom compound plates) to 2x the intended concentration with Tyrode (Sigma) supplemented with 10 mM HEPES (Gibco). The final DMSO concentration in the test solutions and vehicle controls was 0.1%.

Cells hSC-CM (Cor.4U® cardiomyocytes) were obtained from CDI (Ncardia, Germany). Cells were pre-plated and seeded in fibronectin-coated 96-well plates at a suitable density to form a monolayer and maintained in culture in a stage incubator (37° C., 5% CO ₂ ) according to the cell supplier's instructions.

A second lineage of hSC-derived cardiomyocytes, called iCell® Cardiomyocytes2, was purchased from FUJIFILM Cellular Dynamics (USA). Experiments with test drugs are performed 5-7 days after plating the cells on the plate to have a live, beating monolayer of hiPSC-derived cardiomyocytes. Beating monolayers in 96-well plates are typically taken from 2 vials of frozen iCell® Cardiomyocytes2 (≈5 million cells/vial) and plated into three 96-well plates (approximately 50K/well).

At least 1 hour before the start of the experiment, normal cell culture medium was replaced with Tyrode's solution containing a calcium dye (see below).

Cal 520 dye (AAT Bioquest) was dissolved in 11 mL of Tyrode supplemented with 10 mM HEPES and warmed to 37°C before being added to the cells.

35 μl of cell culture medium was removed from each well and replaced with 35 μl of pre-warmed Cal 520 dye solution and the cell plate was incubated for 45 minutes at 37° C./5% CO _2. The cells were incubated at 37° C. for 5 minutes.

Experimental Spontaneous electrical activity is recorded using Cal 520™ (AAT Bioquest) calcium fluorescent dye signaling. This dye integrates total intracellular calcium activity across the well. A bottle of Cal520 dye (50 μg, MW: 1103/mol) is dissolved in 50 μl of DMSO as a 0.9 mM stock solution. 50 μL of the dye stock solution was added to 10 mL of Tryodes solution to give a dye concentration of 4.5 μM. 35 μL of this dye solution was then added to each well to give a final dye concentration of 1.58 μM. The current dye protocol for this CTCM human assay was recently established (Ivan Kopljar et al, Journal of Pharmacological and toxicological methods 2018.91:80-86; Lu et al., Tox Sci 2019.170(2):345-356).

Fluorescence signals (in the form of Ca ²⁺ transients) were measured using a functional drug screening system (FDSS/μCell, Hamamatsu, Japan) and the recordings were subsequently analyzed offline using appropriate software, e.g., Notocord.

For the test run, the cell plate was loaded into the FDSS/μCell and Ca ²⁺ transients were measured for 4 minutes to confirm synchronous beating of cardiomyocytes in each well. All 96 wells were measured simultaneously (sampling interval: 0.06 sec, short exposure time: 10 ms, excitation wavelength 480 nm, emission wavelength 540 nm, FDSS/μCell warmed to 37° C.). If all showed synchronous beating, the 96-well plate was measured in triplicate (wells that did not meet the pre-set criteria to verify synchronous beating of all 96 wells at baseline were excluded from the experiment and not treated with compound).
T=0: control period (-5 to -1 min) plus compound addition, followed by 3 min.
T=30: Measured 29-34 minutes after compound addition

During the compound addition step, 100 μl of each doubly concentrated test solution was pipetted simultaneously into each well.

Data were analyzed offline using appropriate software, e.g. Notocord-Hem (version 4.3).

The following parameters of Ca ²⁺ transient morphology were measured:
- Beat rate (BR)
- Ca2 ⁺ transient amplitude (Amp),
_-CTD90 : duration of Ca2 ⁺ transient at 90% (time to reach 90% of the initial basal value).

A variety of "arrhythmia-like" activities were also noted during the study. These included the following:
• "early afterdepolarization-like" (EAD-like) events (defined as "a very small peak in the transient waveform following the initial peak of the transient");
- a "ventricular tachycardia-like" (VT-like) event (defined as a very fast rate), or - a "ventricular fibrillation-like" (VF-like) event (defined as "small amplitude fast rate Ca2 ⁺ waveforms with irregularities and non-measurable potential transients").
• Cells "stop beating" (no Ca ²⁺ transients are observed).

When compound-induced changes in calcium transient signals could not be resolved by the software, these signals were identified as BQL (below quality analysis level).

Data Analysis: The measured data from FDSS-μCell was copied for offline analysis, analyzed, and uploaded to SPEC-II (our operation management system) for further analysis. The values of the variables before and after the administration of the compound were collected and transferred to Excel workbook.

All values (actual units and percentage change from baseline values) are expressed as median (minimum and maximum). Changes relative to the corresponding baseline values (in actual units) observed in the compound groups were compared to changes in the vehicle control group using the Wilcoxon-Mann-Whitney test. Two-tailed tests with Bonferroni correction for multiplicity adjustment were performed. As there were 10 treatment groups each compared to the vehicle group, an alpha level of 0.05/10 (0.005) was considered to reflect a statistically significant difference from the vehicle group. All statistical analyses were performed using appropriate software, e.g., R software version 3.5.2.

Quality control of hiPSC-CM in plates:
Plates were rejected if they did not meet the following criteria:
-Stable regular beat -Amplitude of more than 500 relative units -Beat rate between 25 and 80 beats/min -CTD ₉₀ of 300 to 800 ms.

In this experiment, the hiPSC-CMs in the plates met the above criteria.

These parameters, combined with the incidence of arrhythmias or beating arrests, were used to calculate potential hazard levels using a weighted scoring method (based on Kopljar et al., Stem Cell Reports 2018.11, 1365-1377). The hazard score is calculated for each concentration by adding weighted points based on the tolerance interval (TI) for changes in CTD ₉₀ , beating rate and amplitude (ΔΔ%), and occurrence of beating arrests and early afterdepolarizations (EADs). As a result, for each concentration, one of four different hazard levels is generated. This is done after 30 minutes of incubation with the compounds. The hazard levels are as follows:
No risk: Effect level or small, insignificant changes in vehicle.
Low Risk: Meaningful effect, but risk of cardiac liability is probably low.
High risk: Relatively high risk for cardiac strain.
Very high risk: Very high risk from arrhythmia-like events (EADs).

The "Hazard Score" results identify potential acute cardiac drug-induced effects at free drug equivalents (as no plasma proteins were added to the wells). The assessment of hazard identification was performed using a scoring reference book called CTCM_Scoring_version1 (Kopljar et al., Stem Cell Reports 2018.11:1365-1377) and indicates the levels according to the following color system in Table 10.

関連する表で異なる色で上に列挙したＨｉＰＳｃ－ＣＭで測定したＣａ^２＋過渡現象アッセイに対するハザードスコア重症度による試験化合物のランク付け。

Ranking of test compounds according to hazard score severity for the Ca ²⁺ transient assay measured in HiPSc-CMs listed above in different colors in the associated table.

Results iCell® Cardiomyocytes 2 was used as the cell line Positive and Negative Controls: All positive and negative controls had the expected pharmacological effect in this assay. The results are summarized in Tables 11 and 12 below.

化合物Ａ１の場合：３０ｍｇ／ｋｇのマウス異種移植モデルにおける有効用量で、ＣＴＣＭヒト濃度対遊離Ｃ_ｍａｘは以下のように推定されるであろう。

For Compound A1: At an effective dose in the mouse xenograft model of 30 mg/kg, the CTCM human concentration vs. free _Cmax would be estimated as follows:

Margin CTCM human 10 μM vs. free Cmax > 16 (mouse, human)
Margin CTCM human 30 μM vs. free Cmax >45 (mouse, human).

8) Effects on membrane potassium current I _Kr in hERG transfected cell lines Protocol 1:

Methods Experiments were performed with CHO cells stably expressing the hERG potassium channel. Cells were grown in culture flasks at 37°C and 5% _CO2 in Ham's F12 medium supplemented with 10% heat-inactivated fetal bovine serum, hygromycin B (100 μg/ml) and geneticin (100 μg/ml). QPatch (Sophion) cells were harvested to obtain a cell suspension of single cells for use in an automated patch clamp system.

Solutions: The bath solution contained 145 mM NaCl, 4 KCl, 10 glucose, 10 HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), 2 CaCl ₂ and 1 MgCl ₂ (pH 7.4 with NaOH). The pipette solution contained 120 mM KCl, 10 mM EGTA (ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid), 10 mM HEPES, 5.374 mM CaCl ₂ and 1.75 mM MgCl ₂ (pH 7.2 with KOH).

Patch clamp experiments were performed in voltage clamp mode and whole-cell currents were recorded in an automated patch clamp assay utilizing the QPatch system (Sophion). Current signals were amplified, digitized, stored, and analyzed using the QPatch assay software.

The holding potential was -80 mV. hERG currents (K ⁺ selective outward currents) were measured as the maximum tail current at -40 mV after a 2 s depolarization to +60 mV. The pulse cycling rate was 15 s. A short pulse (90 ms) to -40 mV served as a baseline step for calculating the tail current amplitude. After establishing the whole-cell configuration and a stable period, a solvent control (0.3% DMSO) was applied for 5 min, followed by the application of test substances at four increasing concentrations: ^3x10-7 M, ^3x10-6 M, ^10-5 M and ^3x10-5 M. Each concentration of test substance was applied twice. Results for each concentration were measured as the mean current of three consecutive voltage pulses after 5 min. Residual currents were compared to vehicle pretreatment to examine the extent of blockade.

Concentration/response relationships were calculated by nonlinear least-squares fitting to the individual data points. Half-maximal inhibitory concentrations (IC50) were calculated by the fitting routine.

Protocol 2:
Cells Compounds, vehicle controls and positive controls were tested in hERG transfected HEK293 cells. A human embryonic kidney cell line (HEK293) with stable transfection of hERG (Zhou Z et al. Biophysical Journal 1998.74,230-241; McDonald T.V. et al, Nature 1997.388,289-292) was used (University of Wisconsin, Madison, USA). Using a T175 flask, cells were kept cultured in MEM (Minimum Essential Medium, Gibco) supplemented with (volumes added to 500 mL MEM are shown): 5 mL L-glutamine-penicillin-streptomycin (Sigma), 50 ml fetal bovine serum (Bio-Whittaker), 5 mL non-essential amino acids 100x (Gibco), 5 ml sodium pyruvate 100 mM (Gibco) and 4 mL geneticin 50 mg/mL (Gibco). Cells were incubated at 37°C in a 5% _CO2 atmosphere (in air).

Cell harvesting for assays Cells were harvested as described below using Accumax™ (Sigma) as a dissociation reagent: Cells were then resuspended in a mixture of 33% DMEM/F12 (Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12-Sigma) medium/67% extracellular physiological solution.

The flasks were carefully washed twice with about 5-10 mL of phosphate buffered saline (PBS) (Gibco™) containing 2 mM EDTA (ethylenediaminetetraacetic acid) (Sigma). The cells were dissociated using about 3 mL of Accumulax™ (cell detachment solution) and incubated at 37°C for about 5-10 minutes. Cold external physiological solution (2-5 mL) is added and the flasks are incubated at about 4°C for 5-10 minutes. The cell suspension in each flask was then gently dissociated with a 5 mL pipette. The cell suspension was transferred to a low-binding Petri dish (diameter about 10 mm). Each flask was further washed with about 5 ml of cold external physiological solution and this solution was also added to the Petri dish. The Petri dish was then incubated at about 4°C for an additional 5-10 minutes. After further gentle dissociation of the cell suspension in the Petri dish, the cells were transferred to a reservoir held on an orbital shaker at 16°C and 200 rpm. The cells were allowed to recover for approximately 20 minutes before performing the experiment.

Compounds: 10 mM solutions of compounds were used and plated in 384-well plates. Aliquots of stock solutions were diluted in recording solution using automated liquid handling (Biomek FXP, final DMSO concentration: 0.03 to 0.3%). A standard range of screening concentrations ranging from 1 μM to 30 μM was used.

A positive control (E-4031) was included within each run to assess the sensitivity of the assay. Table 13 summarizes the extracellular and intracellular solutions used in the experiments. In Table 13, the compositions of the intracellular solutions and extracellular buffer are given in [mM], and "NMDG" means N-methyl-D-glucamine.

Experimental Design The whole-cell patch clamp technique on transfected cells allows the study of ion channels with limited or no interference from other ion channels. The effect of compounds on hERG currents was studied using an automated planar patch clamp system, SyncroPatch 384 PE (Obergrüssberger et al, Journal of Laboratory Automation 2016.21(6), 779-793). All cells were recorded in whole-cell mode of the patch clamp technique. Modules are integrated into a liquid handling pipetting robotic system, Biomek FXP, for cell and compound application, vehicle and positive controls.

Different concentrations of compounds were applied at two cumulatively increasing concentrations of the compound (1 μM and 10 μM, and 3 μM and 30 μM, respectively). hERG currents were measured as the maximum tail current at -30 mV and the percent inhibition relative to the addition of compound or vehicle and positive control was recorded.

After trapping cells in individual holes in the recording tip using tip-fill solution, the seal was enhanced with seal enhancer solution (increased [Ca ²⁺ ]), and the cells were then washed twice with recording solution before being placed in whole-cell mode using a pressure protocol.

After whole-cell mode was achieved, a test pulse was applied for approximately 10 minutes to quantify hERG currents in control conditions. During this control period, vehicle control solution (recording solution containing 0.03% DMSO) was added in triplicate to individual wells. Cumulatively increasing concentrations of vehicle control, compound or positive control were added while the pulse protocol continued. The effects of vehicle, compound and positive control were measured 5 minutes after drug application. Two concentrations of compound were tested per cell.

The use of internal and recording solutions results in a liquid junction potential of approximately 10 mV, and the command voltage step takes this into account.

Electrophysiological measurements: The membrane currents of the cells were measured at different membrane potentials using the patch clamp technique with an automated patch clamp system. The holding potential was −70 mV. The hERG current (K ⁺ selective outward current) was measured as the maximum tail current at −30 mV after a 2 s depolarization to +70 mV (reference 1, 4). The pulse cycling rate was 15 s.

Data Analysis Leak-corrected hERG currents (K ⁺ selective outward currents) were measured as the maximal tail current at -30 mV following a 2 sec depolarization to +70 mV, measured between 2336.3 ms and 3083.6 ms. The median of three current amplitudes was taken at the end of the control period and at the end of each addition of compound, vehicle and positive control to calculate percent inhibition.

QC parameters were set in the SyncroPatch 384PE PatchControl384 software to automatically exclude wells from analysis if values were out of range. QC criteria depended on the type of recording plate (chip). Typically, a 4xChip (medium size hole) was used to record from hERG transfected HEK293 cells. QC criteria 4-6 were set before the first addition of compound. QC criteria 4 and 5 were also set at the end of each compound addition.

QC Criteria and Tolerances:
1. Board check: -500pA to 500pA
2. Contact seal resistance: -100kOhm to 10MOhm
3. Junction potential offset: 0 to 100 mV
4. Rseal≧100 MOhm
5. Series: 1-25 MOhm
6. hERG tail current before compound addition ≧0.2 nA

Each compound was replicated on the same plate in at least 5 wells. Percent inhibition is recorded as the median of at least 2-3 replicates per concentration. Results are summarized in Tables 14 and 15 for Protocol 1 and Protocol 2, respectively.

9) Efficacy Studies in Disseminated OCI-AML3 Model Test Agents and Controls Compound A3 was formulated in 20% hydroxypropyl-β-cyclodextrin (HP-β-CD) and prepared to reach a total volume of 0.2 mL per dose for a 20 g animal (10 mL/kg). Doses were adjusted by individual body weight daily. Working stocks of Compound A3 were prepared once a week for each experiment and stored at 25°C.

Animals Female SCID beige mice (CB17.Cg-PrkdcscidLystbg-J/Crl/-) were used when approximately 6-8 weeks old and weighing approximately 25 g. All animals were allowed to acclimate to and recover from any shipping-related stress for a minimum of 7 days prior to experimental use. Autoclaved water and irradiated food were provided ad libitum and animals were maintained on a 12-hour light/dark cycle. Cages, bedding, and water bottles were autoclaved prior to use and changed weekly. Tissue culture and cell injection reagents are summarized in Table 16 below.

Tumor Models and Cell Culture Methods The human AML cell line OCI-AML3 was cultured in complete culture medium as indicated (MEM alpha + 20% HI-FBS (heat inactivated fetal bovine serum) + 2 mM L-glutamine + 50 ug/ml gentamicin) at 37°C, 5% _CO2 . Cells were harvested during logarithmic growth and resuspended in cold (4°C) MEM ((Minimum Essential Medium) alpha) in serum-free medium.

For the disseminated OCI-AML3 model, each mouse received 5×10 ⁵ cells by IV injection in a total volume of 0.2 mL using a 26-gauge needle.

Experimental Design Compound A3 was administered orally (PO) daily.

Day 0 is the day of tumor cell implantation and the start of the experiment.

In efficacy studies, mice bearing IV OCI-AML3 xenograft tumors were randomly assigned to treatment groups 3 days after tumor cell engraftment. Treatment with vehicle or Compound A3 (at 30, 50, or 100 mg/kg) began on the same day and was administered daily for 28 days.

Animal Monitoring Animals were monitored daily for clinical signs related to either compound toxicity or tumor burden (ie, hind leg paralysis, lethargy, etc.).

Calculations For survival assessment, results were plotted as percentage survival versus days after tumor implantation. Negative clinical signs and/or weight loss of 20% or more were used as surrogate endpoints for death. Kaplan-Meier survival analysis was used to determine median survival. Increasing percent survival (ILS) was calculated as ((median survival of treated group-median survival of control group)/median survival of control group) x 100. Animals that failed to reach surrogate endpoints due to adverse clinical signs (e.g. ulcerated tumors, weight loss, etc.) or deaths unrelated to treatment were excluded for survival assessment. An ILS of 25% or more is considered biologically significant as defined by NCI criteria. (Johnson JI et al. Br J Cancer. 2001.84(10),1424-1431).

Data Analysis Survival and body weight data were graphed using Prism (version 7). Statistical significance for body weight was assessed as described above. Statistical significance was assessed for Kaplan-Meier survival plots comparing therapeutic treatment groups versus appropriate vehicle-treated controls using the log-rank (Mantel-Cox) test in R software version 3.4.2. Differences between groups were considered significant when p-value was ≦0.05.

Survival Kaplan-Meier survival curves are shown in Figure 3. Mice bearing established OCI-AML3 tumors were orally administered Compound A3 at 30, 50, 100 mg/kg in a 20% HP-β-CD formulation daily for a total of 28 days (n=9-10/group). Compound A3-treated groups achieved median survival times of 75.5 days at 30 mg/kg, 58.5 days at 50 mg/kg, and 75 days at 100 mg/kg compared to a median survival time of 38.5 days for the vehicle-treated control group. Compound A3 treatment statistically significantly increased the lifespan of OCI-AML3 tumor-bearing mice by 96.1%, 51.9%, and 94.8% (at 30, 50, and 100 mg/kg dose levels) compared to the lifespan of control mice (p≦0.001). This was biologically significant ILS according to the NCI criteria threshold of 25% or greater ILS (Johnson JI et al. Br J Cancer. 2001.84(10),1424-1431).

10) Efficacy of Menin-MLL Inhibitor in Combination with Venetoclax, Optionally with Azacitidine, in the OCI-AML3 (NPM1c) IV Model in NSG Mice The disseminated OCI-AML3 model described above was also used to test the efficacy of Menin-MLL Inhibitor (formulated as above) in combination with Venetoclax (formulated in polyethylene glycol [PEG]-400/Phosal 50/dimethyl sulfoxide [DMSO]), optionally with Azacitidine (formulated in 10% DMSO), with the following differences: (i) NSG (non-obese diabetic [NOD] scid gamma or NOD.Cg-Prkdc ^scid Il2rg ^tm1Wjl /SzJ) were utilized instead of female SCID beige, (ii) ^1x10 cells were injected into each mouse instead of ^5x10 cells, (iii) compound treatment was initiated 5 days after cell injection instead of 3 days, (iv) instead of a total volume of 0.2 mL (10 ml/kg) per dose, Compound A1 was formulated to reach a volume of 8 mL/kg and compound was administered in the morning, and venetoclax or azacitidine was administered approximately 6-8 hours apart in the afternoon. Treatment of each group with compound in this efficacy study is summarized in Table 17.

Kaplan-Meier survival curves are shown in Figure 4A. The effect on lifespan of 4 weeks of treatment of mice bearing established OCI-AML3 tumors is summarized in Table 18.

有意でない（ｎｓ）と記された場合を除いて、ビヒクル処置対照マウスと比較してｐ＜０．０５。^＊アザシチジン及びベネトクラクスの単剤療法、並びにアザシチジン＋ベネトクラクスの二重併用に対して、ｐ≦０．０５。

p<0.05 compared to vehicle-treated control mice, except where noted as not significant (ns). ^* p≦0.05 versus azacitidine and venetoclax monotherapy and dual combination of azacitidine + venetoclax.

As reflected in Table 18, treatment of mice with Compound A1 in combination with either venetoclax or venetoclax + azacitidine resulted in a statistically significant increase in the life span (ILS) of OCI-AML3 tumor-bearing mice compared to vehicle control treated mice, or azacitidine or venetoclax alone, or the dual combination of venetoclax + azacitidine. Notably, there was no antagonism with the triple combination (Compound A1 + venetoclax and azacitidine) or dual (venetoclax + azacitidine or Compound A1), and efficacy appears to be driven solely by Compound A1.

11) Efficacy of menin-MLL inhibitor in combination with venetoclax, optionally in combination with azacitidine, in the MOLM-13 (KMT2A-r) IV model in NSG mice A similar approach to the disseminated OCI-AML3 model described herein to investigate the efficacy of menin-MLL inhibitor in combination with venetoclax, optionally in combination with azacitidine, was taken, substituting MOLM-13 cells for OCI-AML3 cells.

Tumor Models and Cell Culture Methods The human AML cell line MOLM-13 was cultured in the indicated complete culture medium (RPMI, Roswell Park Memorial Institute [RPMI] 1640 + 10% HI FBS + 2 mM L-glutamine + 50 μg/ml gentamicin) at 37°C, 5% CO2. Cells were harvested during logarithmic growth and resuspended in cold (4°C) serum-free RPMI 1640 medium. Each mouse was injected with 1 x ¹⁰⁵ MOLM-13 cells, and all other parameters were performed as described for the experiment (10).

The treatment of each group with compounds in this efficacy study is summarized in Table 19.

Kaplan-Meier survival curves are shown in Figure 4B. The effect on lifespan of 4 weeks of treatment of mice bearing established MOLM-13 tumors is summarized in Table 20.

有意でない（ｎｓ）と記された場合を除いて、ビヒクル処置対照マウスと比較してｐ＜０．０５。^＊アザシチジン及びベネトクラクスの単剤療法に対して、ｐ≦０．０５。^＊＊化合物Ａ１、アザシチジン及びベネトクラクスの単剤療法、並びにアザシチジン＋ベネトクラクス二重併用に対して、ｐ≦０．０５。

p<0.05 compared to vehicle-treated control mice, except where noted as not significant (ns). ^* p≦0.05 vs. azacitidine and venetoclax monotherapy. ^** p≦0.05 vs. Compound A1, azacitidine and venetoclax monotherapy, and azacitidine + venetoclax dual combination.

As reflected in Table 20, treatment of mice with compound A1 in combination with either venetoclax or venetoclax + azacitidine resulted in a statistically significant increase in the life span (ILS) of MOLM-13 tumor-bearing mice compared to mice treated with venetoclax alone or the dual combination of venetoclax + azacitidine.

(12) In Vitro Growth Protocols (A) Menin-MLL Inhibitors in Combination with Venetoclax and Optionally Azacitidine The effect of Compound A3 in combination with venetoclax or in combination with venetoclax and azacitidine was examined in a proliferation assay using the MOLM-13 (KMT2A-r) cell line. MOLM-13 cells were seeded at 500 cells/well in 96-well plates and exposed to the indicated drugs alone or in combination at the indicated concentrations detailed in Tables 21A and 21B. In particular, for the combination of Compound A3 with venetoclax (without azacitidine), a range of concentrations of Compound A3 (7 points, 4-fold serial dilutions starting at an initial concentration of 1 μM) were combined with venetoclax (6 points, 5-fold serial dilutions starting at an initial concentration of 1 μM). For the combination of Compound A3 with venetoclax and azacitidine, a range of concentrations of Compound A3 (6 points, 4-fold serial dilutions starting at an initial concentration of 1 μM) were combined in a 25:1 ratio with combinations of azacitidine (7 points, 5-fold serial dilutions starting at an initial concentration of 25 μM) and venetoclax (7 points, 5-fold serial dilutions starting at an initial concentration of 1 μM). Compound A3 and azacitidine (when added) were treated on day 0 and venetoclax was added on day 4. MOLM-13 cells were incubated with the indicated drug combination concentrations at 37° C., 5% _CO2 for 8 days (Compound A3 and azacitidine) and 4 days (venetoclax) in quadruplicate.

Spheroid-like growth was monitored in real time by non-invasive live cell imaging using an Incucyte ZOOM live cell imaging system (Essen BioScience) with a 4x objective, and images were acquired on day 8. Confluence as a measure of spheroid size was determined using the integrated analysis tool that is part of the Incucyte ZOOM software "IncuCyte ZOOM 2016B" (Essen BioScience). DMSO content was normalized to 0.3% and confluence from the DMSO wells was used as the baseline response.

Potential combination effects were assessed on day 8 and analyzed for synergy using the R-based Biochemical Intuitive Generalized Loewe (BIGL) model implemented with the best single agent (HSA) null model (Van der Borght 2017). Data from two independent experiments were pooled and analyzed.

Results The dual combination of Compound A3 and venetoclax significantly increased inhibition of cell proliferation compared to Compound A3 or venetoclax monotherapy (see FIG. 5A). As reflected in the contour plot (FIG. 5A) and detailed in Table 21A below, a strong synergistic effect was observed across the concentration range in MOLM-13 cells, with the magnitude of the effect expressed as the difference between the observed and expected outcomes of the combination treatment, with the 95% confidence intervals shown in brackets.

The triple combination of compound A3 with venetoclax and azacitidine significantly increased inhibition of cell proliferation compared to compound A3 monotherapy or the dual combination of venetoclax and azacitidine (see Figure 5B). As reflected in the contour plot (Figure 5B) and detailed in Table 21B below, a strong synergistic effect was observed across a range of concentrations in MOLM-13 cells, with effect magnitude expressed as the difference between observed and expected outcomes of the combination treatment, with 95% confidence intervals shown in brackets.

なお、アスタリスクがない場合は相加効果、拮抗作用は「^＊」、相乗効果は「^＊＊」で示す。拮抗作用と相乗効果は、ｍａｘＲテストに基づく重要なコールと、それぞれ拮抗作用又は相乗効果として解釈されるそれに関連する数値から導かれる。

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

なお、アスタリスクがない場合は相加効果、拮抗作用は「^＊」、相乗効果は「^＊＊」で示す。拮抗作用と相乗効果は、ｍａｘＲテストに基づく重要なコールと、それぞれ拮抗作用又は相乗効果として解釈されるそれに関連する数値から導かれる。

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

(B) Menin-MLL Inhibitor in Combination with Decitabine and Venetoclax The effect of Compound A4 in combination with decitabine compared to combination with decitabine and venetoclax was investigated in proliferation assays using MOLM-13 (KMT2A-r) and OCI-AML3 (NPM1c) cell lines.

Cell Lines AML cell lines MOLM-13 and OCI-AML3 were purchased from DSMZ. MOLM-13 was grown in RPMI medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. OCI-AML3 cells were cultured in 80-90% alpha-MEM (containing ribonucleosides and deoxyribonucleosides) + 10-20% FBS. All cell lines were cultured at 37°C in a 5% CO2 atmosphere.

Cell Titer Glo Assay AML cell lines, MOLM-13 and OCI-AML3 ( ^5x103 cells/well) were seeded in 96-well plates and grown for 6 days in serum (10%)-containing medium in the presence or absence of inhibitors at the indicated concentrations detailed in Tables 22A and 22B below. Proliferation was analyzed by Cell Titer Glo assay using the CellTiter 96 Aqueous One Solution Cell Proliferation Assay (Promega, Madison, WI, USA) according to the manufacturer's instructions. Data are means with standard deviations of 2-4 independent experiments in technical triplicates. Compound A4 and decitabine were added on day 0 and venetoclax (when added) was added on day 4.

Synergy calculations R-based biochemical intuitive generalized Loewe (BIGL) model implemented with the highest single agent (HSA) null model. Specifically, the BIGL methodology was applied to calculate drug-drug interactions (Van der Borght, K., Tourny, A., Bagdziunas, R. et al. BIGL: Biochemically Intuitive Generalized Loewe null model for prediction of the expected combined effect compatible with partial agonism and antagonism. Sci Rep 2013, 113:1311-1323). 7, 17935 (2017), Thas, O., Tourny, A., Verbist, B., Hawinkel, S., Nazarov, M., Mutambanengwe, K., & Bijnens, L. Statistical detection of synergy: New methods and a comparative study. Pharmaceutical Statistics (2021)).

[1073] Synergy matrix results of BIGL analysis of cell line data with HSA as the mean model calculated based on cell metabolic activity using the Cell Titer-Glo assay. Bootstrap confidence intervals are shown. Effect sizes and their confidence intervals are shown. In particular, each data point is based on the p-value and sign of the respective maxR statistic, and the size of the dot reflects the degree of synergy or antagonism corresponding to a graded scale. If the interval includes 0, there is no significant mean effect.

Results Pairwise matrix combinations of decitabine and Compound A4 in the absence or presence of venetoclax were evaluated in MOLM-13 (KMT2A-AF9, FLT3-ITD) and OCI-AML3 (NPM1c AML) cells using a 6-day CellTiter-Glo assay format. Notably, the combination of decitabine and Compound A4, with or without venetoclax, was synergistically cytotoxic in MOLM-13 (KMT2A-AF9, FLT3-ITD) cells as reflected in the contour plots (FIGS. 6A and 6B) and detailed below in Tables 22A and 22B, respectively.

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

Antagonism in OCI-AML3 (NPM1c AML) cells was observed at low doses of decitabine (15 nM) with or without venetoclax, as reflected in the contour plots (Figures 7A and 7B) and detailed below in Tables 23A and 23B, respectively.

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

In the absence of an asterisk, additive effects are indicated with " ^* " and antagonism with " ^** " for synergy. Antagonism and synergy are derived from significant calls based on maxR tests and their associated values that are interpreted as antagonism or synergy, respectively.

Claims (25)

It is a combination,
a therapeutically effective amount of a menin mixed lineage leukemia 1 (MLL) inhibitor of formula (I) or a tautomer or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof;
a therapeutically effective amount of a B-cell lymphoma 2 (BCL-2) inhibitor; and
optionally, a therapeutically effective amount of at least one other anti-neoplastic agent;
The menin-MLL inhibitor of formula (I) has the structure:

having
In the formula, R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety, said 5- or 6-membered monocyclic aromatic ring optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl, wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ may be substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C _1-6 alkyl substituted with _1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) _{2 -C 1-4} alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O _{)-C 1-4} alkyl;
The combination, wherein R ^9a , R ^9b , R ^10a , R ^10b , R ^10c , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl. The menin-MLL inhibitor of formula (I) is compound A:

or a pharma- ceutically acceptable salt or solvate thereof. The menin-MLL inhibitor of formula (I) is compound A4-a:

or a solvate thereof. The combination of any one of claims 1 to 3, wherein the BCL-2 inhibitor is selected from obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax, gambogic acid, venetoclax, and pharma- ceutically acceptable salts or solvates thereof. The combination of claim 4, wherein the BCL-2 inhibitor is venetoclax, or a pharma- ceutically acceptable salt or solvate thereof. The combination according to any one of claims 1 to 5, wherein the at least one other antineoplastic agent is a hypomethylating agent, a DNA intercalating agent, a pyrimidine analogue, a purine analogue, a kinase inhibitor, a CD20 inhibitor, an isocitrate dehydrogenase inhibitor, an immunomodulatory antineoplastic agent or a dihydroorotate dehydrogenase inhibitor. The combination of claim 6, wherein the at least one other antineoplastic agent is a hypomethylating agent. The combination of claim 7, wherein the hypomethylating agent is azacitidine, or a pharma- ceutically acceptable salt or solvate thereof. The combination of claim 1, wherein the menin-MLL inhibitor is compound A or a pharma- ceutical acceptable salt or solvate thereof, the BCL-2 inhibitor is venetoclax or a pharma- ceutical acceptable salt or solvate thereof, and the at least one other antineoplastic agent is a hypomethylating agent. The combination of claim 9, wherein the hypomethylating agent is azacitidine, or a pharma- ceutically acceptable salt or solvate thereof. A pharmaceutical composition comprising the combination according to any one of claims 1 to 10 and a pharma- ceutical acceptable carrier. A combination according to any one of claims 1 to 10, or a pharmaceutical composition according to claim 11, for use as a medicine. A combination according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 11 for use in the prevention or treatment, in particular the treatment, of a hematopoietic disorder. The combination or pharmaceutical composition for use according to claim 13, wherein the hematopoietic disorder is nucleophosmin 1 (NPM1) mutant leukemia or MLL-rearranged leukemia. The combination or pharmaceutical composition for use according to claim 13, wherein the hematopoietic disorder is acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). 1. A method for treating a subject diagnosed with a hematopoietic disorder, comprising administering to the subject:
a therapeutically effective amount of a menin mixed lineage leukemia 1 (MLL) inhibitor of formula (I) or a tautomer or stereoisomeric form thereof, or a pharma- ceutically acceptable salt or solvate thereof;
a therapeutically effective amount of a BCL-2 inhibitor; and
- optionally administering a therapeutically effective amount of at least one other anti-neoplastic agent;
The menin-MLL inhibitor of formula (I) has the structure:

having
In the formula, R ^1a is -C(=O)-NR ^xa R ^xb , Het, or

represents
Het represents a 5- or 6-membered monocyclic aromatic ring containing 1, 2 or 3 nitrogen atoms and optionally a carbonyl moiety, said 5- or 6-membered monocyclic aromatic ring optionally substituted with 1 or 2 substituents selected from the group consisting of C _3-6 cycloalkyl and C _1-4 alkyl;
R ^xa and R ^xb are each independently selected from the group consisting of hydrogen, C _1-4 alkyl, and C _3-6 cycloalkyl;
R ^1b represents F or Cl;
Y ¹ represents -CR ^5a R ^5b -, -O-, or -NR ^5c -;
R ² is selected from the group consisting of hydrogen, halo, C _1-4 alkyl, —O—C _1-4 alkyl, and —NR ^7a R ^7b ;
U represents N or CH;
n1, n2, n3 and n4 are each independently selected from 1 and 2;
X ¹ represents CH and X ² represents N;
^R4 represents isopropyl;
R ^5a , R ^5b , R ^5c , R ^7a and R ^7b are each independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _3-6 cycloalkyl;
R ³ represents -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b or -C _1-6 alkyl-OH or -C _1-6 alkyl-NR ¹¹ -C(=O)-O-C _1-4 alkyl-O-C(=O)-C _1-4 alkyl, wherein each of the C _1-4 alkyl or C _1-6 alkyl moieties in the definition of R ³ may be substituted, independently of one another, with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH and -O-C _1-4 alkyl;
R ^8a and R ^8b are each independently selected from the group consisting of hydrogen, C _1-6 alkyl, -C(=O)-C _1-4 alkyl, -C(=O)-O-C _1-4 alkyl, -C(=O)-NR ^12a R ^12b and C _1-6 alkyl substituted with _1, 2 or 3 substituents each independently selected from the group consisting of -OH, cyano, halo, -S(=O) _{2 -C 1-4} alkyl, -O-C _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O _{)-C 1-4} alkyl;
The method, wherein R ^9a , R ^9b , R ^10a , R ^10b , R ^10c , R ¹¹ , R ^12a , and R ^12b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl. The menin-MLL inhibitor of formula (I) is compound A:

or a pharma- ceutically acceptable salt or solvate thereof. The menin-MLL inhibitor of formula (I) is compound A4-a:

or a solvate thereof. The method according to any one of claims 16 to 18, wherein the BCL-2 inhibitor is selected from venetoclax, obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax, gambogic acid, and pharma- ceutically acceptable salts or solvates thereof. The method of claim 19, wherein the BCL-2 inhibitor is venetoclax, or a pharma- ceutically acceptable salt or solvate thereof. The method of any one of claims 16 to 20, wherein the at least one other antineoplastic agent is a hypomethylating agent, a DNA intercalating agent, a pyrimidine analog, a purine analog, a kinase inhibitor, a CD20 inhibitor, an isocitrate dehydrogenase inhibitor, an immunomodulatory antineoplastic agent, or a dihydroorotate dehydrogenase inhibitor. 22. The method of claim 21, wherein the at least one other antineoplastic agent is a hypomethylating agent. 23. The method of claim 22, wherein the hypomethylating agent is azacitidine, or a pharma- ceutically acceptable salt or solvate thereof. The method according to any one of claims 16 to 23, wherein the hematopoietic disorder is nucleophosmin 1 (NPM1) mutant leukemia or MLL-rearranged leukemia. The method according to any one of claims 16 to 23, wherein the hematopoietic disorder is acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL).

Images