KR20240006631A - combination therapy - Google Patents

Abstract

A therapeutically effective amount of a menin-MLL inhibitor of formula (I), or a pharmaceutically acceptable salt or solvate thereof; and a therapeutically effective amount of a BCL-2 inhibitor; and, optionally, a therapeutically effective amount of at least one other antineoplastic agent. Also disclosed are methods for treating a subject diagnosed with a hematopoietic disorder using such combinations. The compound is represented by the formula (I):
[Formula (I)]
,
In the above formula, R^1a, R^1b, R², R³, R⁴,U,Y^One, X^One, X², n1, n2, n3 and n4 are defined herein.

Description

combination therapy

The present invention provides a therapeutically effective amount of a menin-mixed lineage leukemia 1 (menin-MLL) inhibitor of formula (I), or a pharmaceutically acceptable salt or solvate thereof; and a therapeutically effective amount of a B cell lymphoma 2 (BCL-2) inhibitor; and, optionally, a therapeutically effective amount of at least one other antineoplastic agent; as well as methods of treating a subject diagnosed with a hematopoietic disorder.

Of the 10 million cancer deaths recorded by GLOBOCAN in 2020, 7.1% were due to hematopoietic disorders. Therefore, there is an urgent need for new treatment modalities for hematopoietic disorders, including acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and acute lymphoblastic leukemia (ALL), as detailed below.

AML is a common hematologic malignancy with an incidence increasing from 3:100,000 in young adults to over 20:100,000 in older adults. The overall survival (OS) rate is 40 to 50% for patients under 60 years of age, but only 5% for patients over 60 years of age. The majority of newly diagnosed AML patients are over 60 years of age. In this patient population, standard induction chemotherapy is often not an option because of increased treatment-related mortality due to age and comorbidities. Standard treatment for patients with AML who are not suitable for combination chemotherapy is treatment with hypomethylating agents (azacytidine or decitabine) or low-dose cytarabine. Despite these frontline treatments, the median OS is only approximately 10 months. In all types of AML, disease recurrence is common despite initial treatment response and is the most common cause of death. Standard chemotherapy and allogeneic stem cell transplantation (when used) often fail to eradicate all tumor proliferating cells and select leukemic proliferating subclones that are resistant to chemotherapy. Patients who are refractory to salvage therapy are treated to provide palliation as current treatment options are extremely limited. The median survival time for these patients is 2 months. Additionally, patients with newly diagnosed intermediate or high-risk MDS and those who relapse after standard care have a poor prognosis and a high risk of progression to AML. Therefore, patients with relapsed/refractory (R/R) AML and MDS, newly diagnosed AML patients who cannot receive induction chemotherapy based on age and comorbidities, and newly diagnosed intermediate/high/very high risk MDS. New treatment methods for patients are urgently needed.

ALL is a disseminated hematological malignancy due to impaired differentiation, proliferation and accumulation of lymphoid progenitor cells in the bone marrow and/or extramedullary sites. ALL is the most common childhood acute leukemia, accounting for 12% of all leukemia cases, with an estimated global incidence of 1 to 4.75 per 100,000. ALL accounts for approximately 20% of adult leukemia. Despite the high complete response (CR) rates (80% to 90%) of current treatments, ALL relapses in the majority of adult patients. The 5-year overall survival rate is approximately 30 to 40% for adult and geriatric patients. Therefore, new treatment modalities for relapsed/refractory ALL in adults, especially elderly patients, are urgently needed.

Embodiments of the present invention include a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof; and BCL-2 inhibitors; and optionally at least one other antineoplastic agent.

Embodiments of the invention include BCL-2 inhibitors; and optionally in combination with at least one other antineoplastic agent. It's about.

Embodiments of the present invention relate to novel methods for treating subjects diagnosed with hematopoietic disorders using such combinations. Embodiments of the novel methods include administering to a subject a therapeutically effective amount of a menin-MLL inhibitor as described herein; and a therapeutically effective amount of a BCL-2 inhibitor; and optionally administering a therapeutically effective amount of at least one other antineoplastic agent; wherein the menin-MLL inhibitor is a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof.

Embodiments of the present invention relate to novel methods for treating subjects diagnosed with hematopoietic disorders using such combinations. Embodiments of the novel methods include administering to a subject a therapeutically effective amount of a menin-MLL inhibitor as described herein; and a therapeutically effective amount of a BCL-2 inhibitor; and administering a therapeutically effective amount of at least one other antineoplastic agent; wherein the menin-MLL inhibitor is a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, the 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 pharmaceutically acceptable salt thereof. solvate; and a therapeutically effective amount of venetoclax, or a pharmaceutically acceptable salt or solvate thereof; and administering a therapeutically effective amount of azacytidine or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, the 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 pharmaceutically acceptable salt thereof. solvate; A therapeutically effective amount of venetoclax, or a pharmaceutically acceptable salt or solvate thereof; and administering a therapeutically effective amount of azacytidine or a pharmaceutically acceptable salt or solvate thereof; Here, venetoclax or a pharmaceutically acceptable salt or solvate thereof is administered to the subject before, simultaneously with, or after administration of the menin-MLL inhibitor; Azacitidine or a pharmaceutically acceptable salt or solvate thereof is administered to the subject before, simultaneously with, or after administration of the menin-MLL inhibitor.

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

[Formula (I)]

,

and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, wherein:

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;

wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;

R ⁴ 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 is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl; Here, each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is 1, 2, each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. may be substituted with 2 or 3 substituents;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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.

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

,

,

and solvates thereof.

Those skilled in the art will understand that its "solvate" 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)besylate salt of benzamide You will understand that it refers to .

In certain embodiments, the menin-MLL inhibitor of Formula (I) is ( R )-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy ) Benzamide besylate salt 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-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy ) Benzamide besylate salt or 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-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy ) Benzamide bis-besylate salt 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 bis-besylate salt 0.5- 2.0 equivalents per 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 bis-besylate salt 2.0 equivalents It's about luggage.

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

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

More particularly, 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 bis-besylate salt 2.0 It relates to crystalline form A of the equivalent hydrate.

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

Figure 1 shows 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 bis-besylate hydrate X-ray powder diffraction (XRPD) pattern of crystalline Form A.
Figure 2 depicts a comparison of tumor volume as a function of time for control and treatment groups treated with regimens comprising various amounts of Compound A3 .
Figure 3 depicts a comparison of percent tumor survival (e.g., Kaplan-Meier survival curves) as a function of time for control and treatment groups treated with regimens comprising various amounts of Compound A3 .
Figure 4A shows monotherapy with vehicle, venetoclax, azacitidine or Compound A1 , a dual combination of venetoclax and azacitidine, or a dual combination of Compound A1 and venetoclax, or Compound A1 , venetoclax and Aza. Shown is a comparison of percent survival as a function of time in mice bearing established OCI-AML3 tumors after treatment with the triple combination of cytidine.
Figure 4B shows monotherapy with vehicle, venetoclax, azacitidine or Compound A1 , a dual combination of venetoclax and azacitine, or a dual combination of Compound A1 and venetoclax, or Compound A1 , venetoclax and Aza. A comparison of percent survival as a function of time of mice bearing established MOLM-13 tumors is shown after treatment with the triple combination of cytidine.
Figure 5A is a contour plot for maxR illustrating the effect of Compound A3 in combination with venetoclax on proliferation of MOLM-13 cells in vitro.
Figure 5B is a contour plot for maxR illustrating the effect of Compound A3 in combination with azacitidine and venetoclax on proliferation of MOLM-13 cells in vitro.
Figure 6A is a contour plot for maxR illustrating the effect of Compound A4 in combination with decitabine on proliferation of MOLM-13 cells in vitro.
Figure 6B is a contour plot for maxR illustrating the effect of Compound A4 in combination with decitabine and venetoclax on proliferation of MOLM-13 cells in vitro.
Figure 7A is a contour plot for maxR illustrating the effect of Compound A4 in combination with decitabine on proliferation of OCI-AML3 cells in vitro.
Figure 7B is a contour plot for maxR illustrating the effect of Compound A4 in combination with decitabine and venetoclax on 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. Accordingly, a C _1-6 alkyl group contains 1 to 6 carbon atoms, etc.

As used herein, the term 'C _1-4 alkyl' as a group or part of a group refers to a saturated straight or branched chain hydrocarbon radical having 1 to 4 carbon atoms, such as methyl, ethyl, n -propyl, iso. Represents propyl, n -butyl, s -butyl, t -butyl, etc.

Similarly, as used herein, the term 'C _1-6 alkyl' as a group or part of a group refers to a saturated straight or branched chain hydrocarbon radical having 1 to 6 carbon atoms, such as methyl, ethyl, n - Propyl, isopropyl, n -butyl, s -butyl, t -butyl, n -pentyl, n -hexyl, etc.

As used herein, the term 'C _3-6 cycloalkyl' as a group or part of a group refers to a saturated cyclic hydrocarbon radical having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and Define cyclohexyl.

It will be clear to the person skilled in the art that S(=O) ₂ or SO ₂ represents a sulfonyl moiety.

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

Creatures like -CRR-

It will be clear to those skilled in the art that . An example of such a group is -CR ^5a R ^5b -.

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

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

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

, and It will be understood that it includes, but is not limited to.

If any variable appears more than once in any configuration, each definition is independent.

If any variable appears more than once 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 clear from the context, it refers to one or more hydrogens on the atom or radical indicated in the expression using 'substituted', in particular 1 to 1. It is intended to indicate that 4 hydrogens, more particularly 1 to 3 hydrogens, preferably 1 to 2 hydrogens, more preferably 1 hydrogen, are replaced by a selection from the indicated group, provided that the normal valency is not exceeded. , the substitution produces a chemically stable compound, i.e., a compound that is sufficiently robust to survive the steps of isolation from the reaction mixture to a useful degree of purity (post-reaction isolation, e.g., purification by silica gel chromatography). In certain embodiments, when the number of substituents is not explicitly stated, the number of substituents is one.

Combinations of substituents and/or variables are permissible only if such combinations result in chemically stable compounds. 'Stable compound' in this context is intended to denote a compound that is sufficiently robust to survive the steps of isolation from the reaction mixture to a useful degree of purity (post-reaction isolation, for example purification by silica gel chromatography).

The term 'optionally substituted' will be understood by those skilled in the art to mean that the atom or radical indicated within the expression using 'optionally substituted' may or may not be substituted (which means substituted or unsubstituted, respectively).

When two or more substituents are present on a moiety, where possible and unless otherwise indicated or clear from the context, they may replace a hydrogen atom on the same atom, or they may replace a hydrogen atom on a different atom within the moiety. can do.

'Saturated' in the context of the present invention means 'fully saturated' unless otherwise specified.

Unless otherwise specified or clear from context, the aromatic ring group may be attached to the remainder of the molecule of formula (I) via any available ring carbon atom (C-linked) or nitrogen atom (N-linked). there is.

Unless otherwise specified or clear from the context, aromatic ring groups may optionally be substituted, where possible, on carbon and/or nitrogen atoms, depending on 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 of.” Likewise, 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., cat, dog, primate, or human), more preferably an animal that is or has been the subject of treatment, observation, or experimentation. Hage refers to humans.

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

The term “composition” is intended to include a product comprising the specified ingredients in the specified amounts, as well as any product that results directly or indirectly from a combination of the specified ingredients in the specified amounts.

As used herein, the terms “treatment” and “treating” mean slowing, blocking, arresting or stopping the progression of a disease or improving one or more symptoms thereof, but not necessarily the complete disappearance of all symptoms. It is intended to mean all processes other than doing.

As used herein, a bond shown only as a solid line and not as a solid or dotted wedge bond, or otherwise shown as having a specific configuration (e.g., R , S ) around one or more atoms. Any formula having takes into account each possible stereoisomer, or a mixture of two or more stereoisomers.

Hereinafter and hereinafter, the term “compound(s) of formula (I)” is intended to include tautomers and stereoisomeric forms thereof.

Hereinafter and hereinafter, the term “compound(s) of formula (Z)” is intended to include tautomers and stereoisomeric forms thereof.

Hereinafter or hereinafter, the terms “stereoisomer”, “stereoisomeric form” or “stereochemical isomeric form” 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 mirror images of each other and cannot be superimposed. A 1:1 mixture of a pair of enantiomers is a racemate or racemic mixture.

Atropisomers (or atropisomers) are stereoisomers with specific spatial configurations that result from limited rotation around a single bond due to large steric hindrance. All atropisomeric forms of compounds of formula (I) are intended to be included within the scope of the present invention.

Diastereomers (or diastereomers) are stereoisomers that are not enantiomers, that is, they are not related mirror images. If the compound contains a double bond, the substituent may be in the E or Z configuration.

Substituents on divalent cyclic saturated or partially saturated radicals may have the cis- or trans-configuration; For example, if the compound contains a disubstituted cycloalkyl group, the substituent may be in the cis or trans configuration.

Accordingly, the present invention includes enantiomers, atropisomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers, and mixtures thereof, whenever chemically feasible.

The meaning of all these terms, i.e., enantiomers, atropisomers, diastereomers, racemic E isomers, Z isomers, cis isomers, trans isomers, and mixtures thereof, are known to those skilled in the art.

Absolute conformation is specified according to the Cahn-Ingold-Prelog system. Configuration in asymmetric atoms is specified by R or S. Resolved stereoisomers whose absolute configurations are not known can be designated by (+) or (-) depending on the direction in which they rotate plane polarized light. For example, split enantiomers of unknown absolute configuration can be designated by (+) or (-) depending on the direction in which they rotate plane polarized light.

If a particular stereoisomer is identified, this means that said stereoisomer is substantially free of other stereoisomers, i.e. less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably 5%. means related to less than %, especially less than 2% and most preferably less than 1%. Accordingly, when a compound of formula (I) is designated, for example, as ( R ), this means that the compound is substantially free of the ( S ) isomer; When a compound of formula (I) is designated, for example, as E , this means that the compound is substantially free of the Z isomer; If a compound of formula (I) is specified as cis, for example, this means that the compound is substantially free of trans isomers.

Some of the compounds according to formula (I) may also exist in tautomeric forms thereof. Although not explicitly indicated in formula (I) above, such forms, if present, are intended to be included within the scope of the present invention. In conclusion, a single compound can exist in both stereoisomeric and tautomeric forms.

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

Pharmaceutically acceptable salts, as hereinbefore or hereinafter referred to, are intended to include the therapeutically active, non-toxic acid and base salt forms that the compounds of formula (I), and solvates thereof, can form.

Suitable acids include, for example, mineral acids, such as hydrohalic acids, such as hydrochloric acid or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; Or, organic acids, such as 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, and pamoic acid. Conversely, the salt form can be converted to the free base form by treatment with an appropriate base.

Compounds of formula (I) containing acidic protons and their solvates can also be converted into 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, etc., organic bases such as primary, secondary, and tertiary salts. Secondary aliphatic and aromatic amines, such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n -salts with butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline, and isoquinoline; Includes benzathine, N-methyl-D-glucamine, hydrabamine salts, and salts with amino acids such as, for example, arginine, lysine, etc. Conversely, the salt form can be converted to the free acid form by treatment with acid.

The term “prodrug” refers to oral or parenteral administration, especially oral administration, in an experimentally detectable amount in vivo within a predetermined time (e.g., within a dosing interval of 0.5 to 24 hours, or for example, within 6 to 24 hours). Includes all compounds that are metabolized to the (more) active form within a 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, particularly 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. Modifications are typically accomplished by synthesizing the parent compound with a prodrug substituent. Generally, a prodrug is a compound in which a hydroxyl, amino, sulfhydryl, carboxy or carbonyl group is attached to any group that can be cleaved in vivo to regenerate the free hydroxyl, amino, sulfhydryl, carboxy or carbonyl group, respectively. Includes.

Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxy 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 solvate includes solvent adduct forms as well as salts thereof, which the compounds of formula (I) are capable of forming. Examples of such solvent addition forms are, for example, hydrates, alcoholates, etc.

The compounds of the invention as prepared in the methods described below can be synthesized in the form of mixtures of enantiomers, especially racemic mixtures of enantiomers, which can be separated from each other according to resolution procedures known in the art. Methods for separating enantiomeric forms of compounds of formula (I), and pharmaceutically acceptable salts and solvates thereof, include liquid chromatography using chiral stationary phases. The pure stereochemically isomeric forms can also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, so long as the reaction occurs stereospecifically. Preferably, if specific stereoisomers are desired, the compounds will be synthesized by stereospecific preparation methods. These methods will advantageously use 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 not more than 20% by weight of the other enantiomer. Preferably the product contains at least 90% by weight of one enantiomer and up to 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% by weight of the other enantiomer.

The present invention also includes isotopically-labeled compounds, which are the same as those recited herein, but where one or more atoms have an atomic mass or mass number that is different from the atomic mass or mass number normally found in nature (or The difference is that it is replaced by the most abundant one.

All isotopes and isotopic mixtures of any particular atom or element as specified herein, whether naturally occurring or synthetically produced, in natural abundance or in isotopically enriched form, are of the invention. considered within the category. Exemplary isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³² P, ³³ P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²² I, ¹²³ I, ¹²⁵ I, ¹³¹ I, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, and ⁸² Br. Preferably, the isotope is selected from the group of ² H, ³ H, ¹¹ C, ¹³ C and ¹⁸ F. Preferably, the isotope is selected from the group of ² H, ³ H, ¹¹ C, and ¹⁸ F. More preferably, the isotope is ² H, ³ H or ¹³ C. More preferably, the isotope is ² H or ¹³ C. More preferably, the isotope is ² H. In particular, deuterated compounds and ¹³ C-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 matrix tissue distribution assays. Tritiated ( ³ H) and carbon-14 ( ¹⁴ C) isotopes are useful due to their ease of preparation and detectability. Additionally, substitution with heavier isotopes such as deuterium (i.e. ² H) provides certain therapeutic advantages resulting from greater metabolic stability (e.g. increased in vivo half-life or reduced dosage requirements). may be provided, and may therefore be desirable in some circumstances. Positron emitting isotopes such as ¹⁵ O, ¹³ N, ¹¹ C, and ¹⁸ F are useful in positron emission tomography (PET) studies. PET imaging of cancer is useful in helping to identify and localize tumors, stage the disease, and determine appropriate treatment. Human cancer cells overexpress many 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]). Additionally, 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, such as tablets or capsules, containing one or more compounds described herein may be administered in more than one dosage form at a time, as appropriate. It is also possible to administer the compounds in sustained release formulations.

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

Alternatively, one or more compounds described herein may be administered by inhalation (intratracheally or intranasally) or in the form of a suppository or pessary, or they may be administered as lotions, solutions, creams, ointments, or dispersible powders. It can be applied topically in the form of dusting powder. For example, they may be incorporated into creams that comprise, consist of, or consist essentially of aqueous emulsions of polyethylene glycol or liquid paraffin. They may also contain, consist of or consist essentially of a wax or soft paraffin base, creams to ointments at a concentration of about 1% to about 10% by weight, optionally containing stabilizers and May be introduced with preservatives. Alternative means of administration include transdermal administration using dermal or transdermal patches.

The pharmaceutical compositions (as well as the compounds alone) used in the methods of the invention can also be injected parenterally, for example intracavernosally, intravenously, intramuscularly, subcutaneously, intradermally or intrathecally. In such cases, the composition will 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 sterile aqueous solutions that may contain other substances, such as sufficient salts and simple sugars to render the solution isotonic with blood.

For oral or sublingual administration, the pharmaceutical composition of the present invention can be administered in the form of tablets or lozenges that can be formulated in a conventional manner.

As a further example, a pharmaceutical composition containing as active ingredients at least one compound of formula (I) or a pharmaceutically acceptable salt or solvate, and a BCL-2 inhibitor and, optionally, at least one other antineoplastic agent, comprising: , can be prepared by mixing the compound(s) with a pharmaceutically acceptable carrier, pharmaceutically acceptable diluent, and/or pharmaceutically acceptable excipient according to conventional pharmaceutical compounding techniques. Carriers, excipients, and diluents can take a wide variety of forms depending on the desired route of administration (e.g., oral, parenteral, etc.). Accordingly, for liquid oral preparations such as suspensions, syrups, elixirs and solutions, suitable carriers, excipients and diluents include water, glycols, oils, alcohols, flavoring agents, preservatives, stabilizers, colorants, etc.; For solid oral preparations such as powders, capsules and tablets, suitable carriers, excipients and diluents include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrants, etc. Solid oral preparations may also optionally be coated with substances such as sugars or enteric coated to control the primary sites of absorption and disintegration. For parenteral administration, carriers, excipients and diluents will typically include sterile water, and other ingredients may be added to increase the solubility and preservation of the composition. Injectable suspensions or solutions can also be prepared using aqueous carriers with suitable excipients such as stabilizers and preservatives.

According to certain embodiments, a method of using a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a BCL-2 inhibitor and, optionally, at least one other antineoplastic agent, comprises: A dosage regimen of about 1 to about 4 times per day for an average (70 kg) human being, from about 0.1 mg to about 3000 mg, or any specific amount or range therein, especially from about 1 mg to about 1000 mg, or thereof. Dosage ranges may include any particular amount or range of the active ingredient within; A therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a BCL-2 inhibitor and, optionally, at least one other antineoplastic agent, may be used to treat the diseases, syndromes, conditions, and disorders to be treated. It is clear to those skilled in the art that it will vary depending on.

Embodiments of the present invention relate to methods of using pharmaceutical compositions for oral administration comprising a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, in an amount of from about 1 mg to about 500 mg. . Advantageously, the compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, may be administered in a once daily dosage, or the total daily dosage may be administered twice, three times and four times per day. It may be administered in divided doses.

The optimal dosage of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof to be administered can be readily determined, depending on the specific compound used, the mode of administration, the strength of the agent, and the progression of the hematopoietic disorder. It will vary depending on the degree. Additionally, depending on factors associated with the particular subject being treated, including subject sex, age, weight, diet, and time of administration, the dosage will need to be adjusted to achieve an appropriate therapeutic level and desired therapeutic effect. Therefore, the above dosages are examples of average cases. Of course, there may be individual cases where a larger or smaller dosage range would be beneficial, and such cases are within the scope of the present invention.

Whenever the use of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, is to be administered to a subject in need thereof, the compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, It can be administered by any of the compositions and administration regimens described above, or by compositions and administration regimens established in the art.

Embodiments of the invention relate to methods of using pharmaceutical compositions 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 may be administered in a single daily dose, or the total daily dose may be administered in divided doses twice, three and four times daily.

The optimal dosage of BCL-2 inhibitor to be administered can be readily determined and will depend on the specific compound used, the mode of administration, the potency of the agent, and the progression of the disease, syndrome, condition or disorder. Additionally, depending on factors associated with the particular subject being treated, including subject sex, age, weight, diet, and time of administration, the dosage will need to be adjusted to achieve an appropriate therapeutic level and desired therapeutic effect. Therefore, the above dosages are examples of average cases. Of course, there may be individual cases where a larger or smaller dosage range would be beneficial, and such cases are within the scope of the present invention.

Whenever use of a BCL-2 inhibitor is to be administered to a subject in need thereof, the BCL-2 inhibitor may be administered in any of the compositions and administration regimens described above, or by compositions and administration regimens established in the art. there is.

As used herein, the term "menin-MLL inhibitor" refers to a protein-protein interaction between menin and mixed lineage leukemia 1 (MLL1) in the scientific field of inhibiting or reducing menin-MLL 1 activity (UniProt Accession # Q03164). Refers to an inhibitor of the interaction (also known herein as histone-lysine N-methyltransferase 2A (KMT2A) protein). The menin-MLL inhibitors described herein are disclosed in PCT/CN2020/137266 (published as WO 2021/121327 on June 24, 2021), which is incorporated herein by reference in its entirety and the corresponding synthesis Reaction schemes and analytical features are also disclosed.

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

As used herein, the term “anti-neoplastic agent” refers to any agent that treats cancer.

As used herein, the term “hypomethylating agent” refers to an agent that inhibits or reduces DNA methylating agents.

As used herein, the term "kinase inhibitor" refers to an inhibitor of at least one kinase (e.g., a tyrosine and/or serine kinase, e.g., fms-like receptor tyrosine kinase-3 (FLT3), Bruton's tyrosine kinase ( BTK), Abelson tyrosine kinase 1 (ABL), Aurora serine/tyrosine kinase).

As used herein, the term "FLT-3 inhibitor" refers to fms-like receptor tyrosine kinase-3 (FLT3), a tyrosine kinase classified as first- and next-generation inhibitors based on their potency and specificity against its associated downstream targets. Refers to an inhibitor (TKI).

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 an inhibitor that inhibits the conversion of isocitrate to α-ketoglutarate (α-KG) in the tricarboxylic acid (TCA) cycle. refers to any agent that

As used herein, the term “immunomodulatory antineoplastic 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 “affecting” or “affected” (when referring to a disease, disorder or medical condition that is affected by inhibition or alteration of menin-MLL activity) ) comprises a reduction in the frequency and/or severity of one or more symptoms or manifestations of said hematopoietic disorder; and preventing the onset of one or more symptoms or manifestations of the hematopoietic disorder or the development of the hematopoietic disorder.

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

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

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

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

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

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

According to one embodiment, the invention provides a combination as described herein for use in the prevention or treatment, particularly treatment, of hematopoietic disorders, including but not limited to hematologic malignancies, including but not limited to lymphoma, myeloma and leukemia. Provide water.

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

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 disease lymphoma and non-Hodgkin's lymphoma.

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

According to one embodiment, the hematopoietic disorder is myeloma. According to 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, myelocytic leukemia, lymphoblastic leukemia, or lymphocytic leukemia. According to one embodiment, the hematopoietic disorder is acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), small lymphocytic leukemia (SLL), acute myeloid leukemia (AML), chronic idiopathic myelofibrosis (MF), chronic myeloid leukemia Constitutive 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 lymphocyte selected from, but not limited to, leukemia (T-LGL) and aggressive NK cell leukemia. According to one embodiment, the AML is acute megaloblastic 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 cancer 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 with an increased HOX/MEIS1 gene expression signature.

According to one embodiment, the leukemia is MLL rearrangement leukemia and/or nucleophosmin 1 (NPM1) mutant leukemia. According to one embodiment, the hematopoietic disorder is MLL rearrangement leukemia.

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

According to one embodiment, the invention provides a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a BCL-2 inhibitor, and optionally at least one other antineoplastic agent. Myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), small lymphocytic lymphoma (SLL), comprising administering to a subject in need thereof. Alternatively, a method of treating a hematopoietic disorder, such as chronic lymphocytic leukemia (CLL), is provided.

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).

According to one embodiment, the hematopoietic disorder is SLL or CLL, wherein the 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 myeloproliferative neoplasm (MPN).

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

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

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

According to one embodiment, the hematopoietic disorder has one or more MLL1 (KMT2A) gene rearrangements or alterations (e.g. duplication or amplification) and/or NPM1 mutations.

According to one embodiment, the hematopoietic disorder has (i) one or more MLL1 (KMT2A) gene rearrangements or alterations (e.g. duplication or amplification) and/or an NPM1 mutation plus (ii) a FLT3 mutation.

According to one embodiment, the hematopoietic disorder is MLL rearrangement 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, myelocytic 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 with an increased HOX/MEIS1 gene expression signature.

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

According to one embodiment, the hematopoietic disorder is MLL rearranged leukemia, especially MLL rearranged AML or ALL.

According to one embodiment, the hematopoietic disorder comprises MLL gene alterations, and in particular the hematopoietic disorder is AML or ALL with MLL gene alteration(s). In certain embodiments, the MLL gene alteration is redundant. In certain embodiments, the MLL gene alteration is an amplification.

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

According to one embodiment, MLL1 gene mutations include, but are not limited to, MLL1 gene rearrangements, duplications, or amplifications.

According to one embodiment, the hematopoietic disorder is mixed lineage leukemia (MLL), MLL-related leukemia, MLL-related leukemia, MLL-positive leukemia, MLL-induced leukemia, leukemia associated with MLL, acute leukemia, chronic leukemia, myelodysplastic syndrome (MDS), or It is a myeloproliferative neoplasm (MPN).

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

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

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

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

In one embodiment, the present invention is a novel combination comprising:

치료적 유효량의 화학식 (I)의 메닌-MLL 억제제, 또는 이의 호변이성질체 또는 입체이성질체 형태, 또는 이의 약제학적으로 허용되는 염 또는 용매화물;

A therapeutically effective amount of a menin-MLL inhibitor of formula (I), or a tautomeric or stereoisomeric form thereof, or a pharmaceutically 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 antineoplastic agent.

According to one embodiment, the compound of formula (I) is a menin-MLL inhibitor having the structure:

[Formula (I)]

,

and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, wherein:

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;

wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;

R ⁴ 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 is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl; Here, each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is 1, 2, each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. may be substituted with 2 or 3 substituents;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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.

According to one embodiment,

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;

wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;

R ⁴ 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 is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl;

wherein each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is independently selected from the group consisting of cyano, halo or -OC _1-4 alkyl, 1, 2 or may be substituted with 3 substituents;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents;

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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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;

wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;

R ⁴ 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 ; Here, the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, OH and -OC _1-4 alkyl;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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;

wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;

R ⁴ 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 ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents;

R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

R ^1a represents -C(=O)-NR ^xa R ^xb or Het;

Het represents a 6-membered monocyclic aromatic ring containing 2 nitrogen atoms; wherein the 6-membered monocyclic aromatic ring is 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-;

R ² represents hydrogen;

U represents N or CH;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl;

Wherein each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ has 1, 2 or 3 substituents each independently selected from the group consisting of -OH and -OC _1-4 alkyl. may be replaced with;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

R ^1a represents -C(=O)-NR ^xa R ^xb or Het;

Het represents a 6-membered monocyclic aromatic ring containing 2 nitrogen atoms; wherein the 6-membered monocyclic aromatic ring is 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-;

R ² represents hydrogen;

U represents N or CH;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; Here, the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH and -OC _1-4 alkyl;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O) -C _1-4 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of 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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N or CH;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; Here, the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -OH and -OC _1-4 alkyl;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O) -C _1-4 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of 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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with one -OH;

R ^8a and R ^8b are each independently 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, -OC _1-4 alkyl, and -NR ^10c -C( ₌ O)-C _1-4 alkyl. selected from the group;

R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n2 is 2;

n1, n3 and n4 are 1;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with one -OH;

R ^8a and R ^8b are each independently 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, -OC _1-4 alkyl, and -NR ^10c -C( ₌ O)-C _1-4 alkyl. selected from the group;

R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n2 is 2;

n1, n3 and n4 are 1;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ;

R ^8a and R ^8b are each independently 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, -OC _1-4 alkyl, and -NR ^10c -C( ₌ O)-C _1-4 alkyl. selected from the group;

R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n2 is 2;

n1, n3 and n4 are 1;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ;

R ^8a and R ^8b are each independently 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, -OC _1-4 alkyl, and -NR ^10c -C( ₌ O)-C _1-4 alkyl. selected from the group;

R ^10a , R ^10b , and R ^10c are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b each independently selected from the group consisting of selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents;

R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b each independently selected from the group consisting of selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents;

R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b each independently selected from the group consisting of selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents;

R ^10a and R ^10b are each independently selected from the group consisting of hydrogen and C _1-6 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ;

R ^8a and R ^8b are each independently 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 -OC _1-4 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ;

R ^8a and R ^8b are each independently C _1-6 alkyl; and C _1-6 alkyl substituted with one -OC _1-4 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² represents hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ;

R ^8a and R ^8b are each independently C _1-6 alkyl; and C _1-6 alkyl substituted with one -OC _1-4 alkyl,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

R ^1a is -C(=O)-NR ^xa R ^xb ; or Het;

Het represents a 6-membered monocyclic aromatic ring containing 2 nitrogen atoms; wherein the 6-membered monocyclic aromatic ring is 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-;

R ² is hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ represents isopropyl;

R ³ is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl;

R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and C _1- 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 -OC _1-4 alkyl. ₆ selected from the group consisting of 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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

According to one embodiment,

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-;

R ² is hydrogen;

U represents N;

n1, n2, n3 and n4 are each independently selected from 1 and 2;

X ¹ represents CH, X ² represents N;

R ⁴ 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 hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and C _1- 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 -OC _1-4 alkyl. ₆ selected from the group consisting of 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,

Compounds of formula (I), and tautomeric and stereoisomeric forms thereof, and pharmaceutically acceptable salts and solvates thereof, are as defined herein.

In one embodiment, the invention provides compounds of formula (I), and pharmaceutically acceptable salts and solvates thereof, wherein R ^1b represents F, or any of the compounds thereof as mentioned in any of the other embodiments. Includes subgroups.

In one embodiment, the invention provides compounds of formula (I), and pharmaceutically acceptable salts and solvates thereof, wherein R ² represents hydrogen, or any of the compounds thereof as recited in any of the other embodiments. Includes subgroups.

In one embodiment, the invention provides compounds of formula (I), wherein n1 is 1, n2 is 2, n3 is 1, and n4 is 1, and pharmaceutically acceptable salts and solvates thereof, or other and any subgroup thereof as mentioned in any of the embodiments.

In one embodiment, the present invention provides compounds of formula (I), and pharmaceutically acceptable salts and solvates thereof, wherein Y ¹ represents -O-, or a compound thereof as recited in any of the other embodiments. Includes any subgroup.

In one embodiment, the invention provides that Y ¹ represents -O-; U represents N, and includes compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that Y ¹ represents -O-; U represents N; R ^1b represents F; R ² represents hydrogen, including compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the present invention provides Het

Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that Het represents a monocyclic 5- or 6-membered aromatic ring containing 1 or 2 nitrogen atoms;

wherein the monocyclic 5- or 6-membered aromatic ring is substituted with one C _3-6 cycloalkyl, and pharmaceutically acceptable salts and solvates thereof, or in other embodiments It includes any subgroup thereof as mentioned in any one.

In one embodiment, the invention provides that Het represents a monocyclic 5- or 6-membered aromatic ring containing 1 or 2 nitrogen atoms; Here, the monocyclic 5- or 6-membered aromatic ring is substituted with one C _3-6 cycloalkyl; R ^1b includes compounds of formula (I), which represents F, and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that Het represents a monocyclic 6-membered aromatic ring containing 1 or 2 nitrogen atoms; wherein the monocyclic 6-membered aromatic ring is substituted with one C _3-6 cycloalkyl, and pharmaceutically acceptable salts and solvates thereof, or in any of the other embodiments. Includes any subgroups thereof as mentioned.

In one embodiment, the invention provides that Het represents a monocyclic 6-membered aromatic ring containing 1 or 2 nitrogen atoms; Here, the monocyclic 6-membered aromatic ring is substituted with one C _3-6 cycloalkyl; R ^1b includes compounds of formula (I), which represents F, and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety as defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; ) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; Here, the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. , compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides compounds of formula (I), and pharmaceutically acceptable salts and solvates thereof, or in other embodiments, wherein R ³ represents -C _1-6 alkyl-NR ^8a R ^8b It includes any subgroup thereof as mentioned in any one.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O)-C _1-4 Compounds of formula (I) selected from the group consisting of alkyl substituted with 1, 2 or 3 substituents each independently selected from _the group consisting of alkyl and pharmaceuticals thereof salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O)-C _1-4 Compounds of formula (I) selected from the group consisting of alkyl substituted with 1, 2 or 3 substituents each independently selected from _the group consisting of alkyl and pharmaceuticals thereof salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents, or as mentioned in any of the other embodiments It includes any subgroup thereof as defined.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)- Compounds of formula (I) and pharmaceutically acceptable compounds thereof selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of C _1-4 alkyl salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents, or as mentioned in any of the other embodiments It includes any subgroup thereof as defined.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; wherein the C _1-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)- Compounds of formula (I) and pharmaceutically acceptable compounds thereof selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of C _1-4 alkyl salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety as defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; ) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; Here, the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. , compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O)-C _1-4 Compounds of formula (I) selected from the group consisting of alkyl substituted with 1, 2 or 3 substituents each independently selected from _the group consisting of alkyl and pharmaceuticals thereof salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O)-C _1-4 Compounds of formula (I) selected from the group consisting of alkyl substituted with 1, 2 or 3 substituents each independently selected from _the group consisting of alkyl and pharmaceuticals thereof salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents, or as mentioned in any of the other embodiments It includes any subgroup thereof as defined.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and one independently selected from the group consisting of cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b , Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, selected from the group consisting of C _1-6 alkyl substituted with 2 or 3 substituents, or as mentioned in any of the other embodiments It includes any subgroup thereof as defined.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ;

R ^8a and R ^8b are each independently C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, and -C(=O)-NR ^10a R ^10b each independently selected from the group consisting of Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any of the other embodiments, selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents. It includes any subgroup thereof as mentioned above.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl; ; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)- Compounds of formula (I) and pharmaceutically acceptable compounds thereof selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of C _1-4 alkyl salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _2-6 alkyl-NR ^8a R ^8b ; wherein the C _2-6 alkyl moiety defined by R ³ may be substituted with 1, 2 or 3 substituents each independently selected from the group consisting of cyano, halo and -OC _1-4 alkyl; R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; and cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(=O)- Compounds of formula (I) and pharmaceutically acceptable compounds thereof selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of C _1-4 alkyl salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ;

R ^8a and R ^8b are each independently C _1-6 alkyl; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b and -NR ^10c -C(=O )-C _1-4 alkyl, each independently selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents, acceptable salts, and solvates, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ represents -C _1-6 alkyl-NR ^8a R ^8b ; R ^8a represents C _1-6 alkyl; R ^8b refers to any of the compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or other embodiments, wherein R 8b represents C _1-6 alkyl substituted with one -OC _1-4 alkyl. Includes any subgroup thereof as defined.

In one embodiment, the invention provides that R ³ is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl; wherein each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is independently selected from the group consisting of cyano, halo or -OC _1-4 alkyl, 1, 2 or Compounds of formula (I), which may be substituted with three substituents, and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides that R ³ is -C _1-6 alkyl-NR ^8a R ^8b , -C _1-6 alkyl-C(=O)-NR ^9a R ^9b , or -C _1-6 alkyl- NR ¹¹ -C(=O)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl; Here, each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is 1, 2, each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. Compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, which may be substituted with 2 or 3 substituents, or any subgroup thereof as mentioned in any of the other embodiments. .

In one embodiment, the present invention provides compounds of formula (I), wherein R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b and pharmaceutically acceptable salts and solvates thereof, or other and any subgroup thereof as mentioned in any of the embodiments.

In one embodiment, the invention provides that R ³ represents -CH ₂ -CH ₂ -CH ₂ -NR ^8a R ^8b ; R ^8a represents methyl; R ^8b represents -CH ₂ -CH ₂ -OCH ₃ , compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof, or any subtype thereof as mentioned in any of the other embodiments. Includes military.

In one embodiment, the invention provides a method of formula (I) wherein the C _1-6 alkyl moiety of R ³ defined -C _1-6 alkyl-NR ^8a R ^8b is limited to -CH ₂ -CH ₂ -CH ₂ - Compounds and pharmaceutically acceptable salts and solvates thereof, or any subgroup thereof as mentioned in any of the other embodiments.

In one embodiment, the invention provides a compound of formula (I) and a pharmaceutically acceptable salt or solvate thereof, wherein the compound of formula (I) is limited to a compound of formula (Ia) or formula (Ib), or Includes any subgroup thereof as recited in any of the other embodiments:

(Ia) 또는

(Ib),

(Ia) or

(Ib),

where R ^1a , R ^{1b , R 3} , R ⁴ , R ^5a , R ^5b , X 1 , As defined for any subgroup thereof as mentioned above.

In one embodiment, the compounds of formula (I) and pharmaceutically acceptable salts or solvates thereof, or any subgroup thereof as recited in any of the other embodiments, are compounds of formula (Ia), or It is limited to pharmaceutically acceptable salts or solvates thereof. In one embodiment, the compounds of formula (I) and pharmaceutically acceptable salts or solvates thereof, or any subgroup thereof as recited in any of the other embodiments, are compounds of formula (Ib), or It is limited to pharmaceutically acceptable salts or solvates thereof.

In one embodiment, the invention provides a compound of formula (I) and a pharmaceutically acceptable salt or solvate thereof, wherein the compound of formula (I) is limited to a compound of formula (I-y), or any of the other embodiments. Includes any subgroup thereof as mentioned in:

[Formula (I-y)]

,

wherein R ³ is as defined for the compound of formula (I) or any subgroup thereof as mentioned 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 or a pharmaceutically acceptable salt or solvate thereof:

Compound A

In a preferred embodiment, the compound of formula (I) is compound A1 :

Compound A1

.

In a preferred embodiment, the compound of formula (I) is compound A2 :

Compound A2

.

In a preferred embodiment, the compound of formula (I) is compound A3 :

Compound A3

.

.

In certain embodiments, the compound of Formula (I) is Compound A4-a or a solvate thereof:

Compound A4-a

In certain embodiments, the menin-MLL inhibitor of Formula (I) is ( R )-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy ) Benzamide besylate salt 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-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy )benzamide bis-besylate salt or 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-meth Toxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy )Benzamide bis-besylate salt (Compound A4-b ) or a hydrate thereof.

In certain embodiments, the menin-MLL inhibitor of Formula (I) is 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-triazine-6- It is crystalline form A of 1)oxy)benzamide bis-besylate salt hydrate.

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

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

In one embodiment, the compound of formula (I) is selected from the group consisting of any of the exemplified compounds, tautomeric and stereoisomeric forms thereof, and free bases, any pharmaceutically acceptable salts, and solvates thereof. do.

In some embodiments, a pharmaceutical composition is provided comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the combination as an active ingredient as described in any of the other embodiments.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor and, optionally, at least one other antineoplastic agent. .

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

According to certain embodiments, the menin-MLL inhibitor is Compound A or a pharmaceutically acceptable salt or solvate thereof.

According to a specific embodiment, the menin-MLL inhibitor is Compound A1 .

According to a specific embodiment, the menin-MLL inhibitor is Compound A2 .

According to a specific embodiment, the menin-MLL inhibitor is Compound A3 .

According to certain embodiments, the menin-MLL inhibitor is Compound A4-a or a solvate thereof.

According to certain embodiments, the menin-MLL inhibitor is Compound A4-b or a hydrate thereof.

According to a particular embodiment, the menin-MLL inhibitor is Compound A4 .

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

In certain embodiments, the menin-MLL inhibitor 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 can reduce tumor growth, e.g., 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 long periods of time, such as inhibition of target gene expression such as MEIS1 and upregulation of differentiation markers over 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 is Q.D. May be suitable for dosing (once daily).

According to an embodiment, the BCL-2 inhibitor is obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax, gamgogic acid and venetoclax, or a pharmaceutically acceptable salt or solvate thereof.

According to certain embodiments, the BCL-2 inhibitor is venetoclax, or a pharmaceutically acceptable salt or solvate thereof.

According to an embodiment, 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 an embodiment, 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 (IDH) inhibitor.

According to an embodiment, the hypomethylating agent includes, but is not limited to, azacitidine, decitabine, or a pharmaceutically acceptable salt or solvate 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 an embodiment, pyrimidine analogs include, but are not limited to, cytarabine (ARA-C).

According to an embodiment, the purine analog is fludarabine.

According to embodiments, the kinase inhibitor is a multi-kinase inhibitor of a FLT-3 inhibitor, a BTK inhibitor, an ABL inhibitor, an Aurora inhibitor, or two or more kinase inhibitors thereof.

According to an embodiment, the kinase inhibitor is a multi-kinase inhibitor of a FLT-3 inhibitor, an ABL inhibitor, and an Aurora inhibitor. According to embodiments, such multi-kinase inhibitors include, but are 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 a FLT-3 inhibitor or a BTK inhibitor.

According to an embodiment, the FLT3 inhibitors include sorafenib, sunitinib, midostaurin (PKC412), lestaurtinib (CEP-701), tandutinib (MLN518), quizartinib (AC220), gilteritinib (ASP2215), and Includes but is not limited to KW-2449.

According to an embodiment, the BTK inhibitor includes, but is not limited to, ibrutinib.

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

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

According to an embodiment, the isocitrate dehydrogenase-1 inhibitor includes, but is not limited to, ivosidenib.

According to an embodiment, the isocitrate dehydrogenase-2 inhibitor includes, but is not limited to, enasidenib.

및 레바미솔을 포함하지만 이로 한정되지 않는다.According to an embodiment, the immunomodulatory antineoplastic agent is a PD-1 inhibitor (e.g., nivolumab, atezolizumab, and pembrolizumab), thalidomide, lenalidomide, pomalidomide, Calmette-Guerin.

and levamisole.

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

According to an embodiment, the DHODH inhibitor is a compound having the structure of Formula (Z):

[Formula (Z)]

,

(In the above equation

X is CH or N;

Y is CH or N;

R ¹ is 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;

R ² is ego; here

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;

R ³ is selected from the group consisting of H, halo, CH ₃ , and OCH ₃ ;

R ⁴ 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 ;

;

and ; and

, and is selected from the group consisting of;

here,

Each R ^c is independently 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 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 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 pharmaceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof;

or a compound selected from:

,

or pharmaceutically acceptable salts, N-oxides, solvates, or stereoisomers thereof.

According to an embodiment, the DHODH inhibitor includes, but is not limited to, a compound having the structure of Formula (Z), or a pharmaceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof:

[Formula (Z)]

,

In the above equation

X is CH or N;

Y is CH or N;

R ¹ is 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;

R ² is ego; here

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;

R ³ is selected from the group consisting of H, halo, CH ₃ , and OCH ₃ ;

R ⁴ 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 ;

;

and ; and

, and is selected from the group consisting of;

here,

Each R ^c is independently 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 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 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.

In the context of formula (Z), the following definitions apply:

The term “alkenyl” includes unsaturated aliphatic groups similar in length and possible substitution to the alkyls described above, but containing at least one double bond. For example, the term “alkenyl” includes straight chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.). The term alkenyl further includes alkenyl groups containing oxygen, nitrogen, sulfur or phosphorus atoms replacing one or more carbons of the hydrocarbon backbone. In certain embodiments, a straight or branched chain alkenyl group has no more than 6 carbon atoms in its backbone (eg, C _2-6 for straight chain, C _3-6 for branched chain).

The term “haloalkyl” refers to a straight or branched chain alkyl group having from 1 to 6 carbon atoms in the chain, wherein hydrogen is optionally replaced by halogen. As used herein, the term “C _1-6 haloalkyl” refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms in the chain, with hydrogen optionally substituted with halogen. As used herein, the term “C _1-4 haloalkyl” refers to a straight or branched chain alkyl group having from 1 to 4 carbon atoms in the chain, with hydrogen optionally substituted with halogen. Examples of “haloalkyl” groups include trifluoromethyl (CF ₃ ), difluoromethyl (CF ₂ H), monofluoromethyl (CH ₂ F), pentafluoroethyl (CF ₂ CF ₃ ), tetrafluoro. Roethyl (CHFCF ₃ ), monofluoroethyl (CH ₂ CH ₂ F), trifluoroethyl (CH ₂ CF ₃ ), tetrafluorotrifluoromethylethyl (CF(CF ₃ ) ₂ ) and the present technical field. In consideration of the common skill and teachings given herein, it includes groups that are considered equivalent to any of the examples mentioned above.

The term "haloalkenyl" refers to an unsaturated aliphatic group similar in length and possible substitution to the alkyl described above, but having from 1 to 6 carbon atoms in the chain containing at least one double bond and optionally replacing hydrogen with halogen. Includes.

The term “aryl” refers to a monocyclic, aromatic carbon ring having six atoms per ring (a ring structure in which all ring atoms are carbon). (The carbon atoms of the aryl group are sp2 hybridized.)

The term “heteroaryl” refers to a monocyclic or fused bicyclic heterocycle (a ring selected from a carbon atom and up to 4 heteroatoms selected from nitrogen, oxygen and sulfur), having 3 to 9 ring atoms per heterocycle. refers to a ring structure with atoms). Illustrative examples of heteroaryl groups include the following entities in the form of appropriately linked moieties:

Those skilled in the art will recognize that the chemical species listed or exemplified above are not exhaustive and that additional chemical species may also be selected within the scope of these defined terms.

The term “variable point of attachment” means that any group can be attached to more than one alternative position within the structure. Attachment will always replace a hydrogen atom from one of the ring atoms. In other words, all permutations of bonds are represented in a single diagram, as shown in the examples below.

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein X is CH.

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

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein Y is CH.

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein Y is N.

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

인 화학식 (Z)의 화합물이다.In an embodiment of the invention, the DHODH inhibitor is R ¹

Phosphorus is a compound of formula (Z).

In an embodiment of the invention, the DHODH inhibitor is

R ² ego;

where 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;

and R ^a is C _1-4 alkyl, C _1-4 haloalkyl, or C _3-6 cycloalkyl.

In an embodiment of the invention, the DHODH inhibitor has R ² Phosphorus is a compound of formula (Z).

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

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

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

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

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein R ⁴ is:

In an embodiment of the invention, the DHODH inhibitor is

R ⁴ a ego; here

Each R ^c is independently 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 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;

A compound of formula (Z) where n is 1 or 2.

In an embodiment of the invention, the DHODH inhibitor is

R ⁴ a ego;

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;

A compound of formula (Z) where n is 1 or 2.

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein R ⁴ is:

, or .

In an embodiment of the invention, the DHODH inhibitor has R ⁴ or Phosphorus is a compound of formula (Z).

In an embodiment of the invention, the DHODH inhibitor is

R ⁴ a , or and;

here,

Each R ^c is independently 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 ; 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; C _1-4 haloalkyl substituted with OH or OCH ₃ ; or OC _1-4 alkyl; CN; and OC _1-6 alkyl.

In an embodiment of the invention, the DHODH inhibitor is

R ⁴ a or and; here

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;

A compound of formula (Z) where n is 1 or 2.

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) wherein R ⁴ is:

, or .

In an embodiment of the invention, the DHODH inhibitor has R ⁴ Phosphorus is a compound of formula (Z).

In an embodiment of the invention, the DHODH inhibitor is

R ⁴ a , or and; here

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 an embodiment of the invention, the DHODH inhibitor is

R ⁴ a , or and; here

R ^c is H or halo;

R ^d is C _1-4 alkyl;

R ^g is a compound of formula (Z) wherein is H.

인 화학식 (Z)의 화합물이다.In an embodiment of the invention, the DHODH inhibitor has R ⁴

Phosphorus is a compound of formula (Z).

In an embodiment of the invention, the DHODH inhibitor is a compound 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)-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-triazole -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-triazole -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-triazole -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-triazole-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-triazole-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-triazole -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-Isopropylphthalazine-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-triazole -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-iso Propyl-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-triazole -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-triazole -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-triazole -1-yl)-7-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;

2-(5-chloro-3-methyl-1 H -pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- (prop-1-en-2-yl)-6-( o -tolyl)-1,6-naphthyridin-5(6 H )-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- methyl-6-( o -tolyl)pyrido[2,3-d]pyridazin-5(6 H )-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-tri Azol-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-triazole -1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;

(S)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluo Ro-6-( o -tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6 H )-one;

(R)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluo Ro-6-( o -tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6 H )-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- Isopropyl-6-( o -tolyl)-1,6-naphthyridin-5(6 H )-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-triazole-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-triazole -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-iso Propyl-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-triazole -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-iso Propyl-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-triazole -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-fluo Ro-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-fluo Ro-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-triazole -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-methoxyphenyl)-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-triazole- 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-triazole -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-triazole- 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-( 4-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-triazole-1- 1)-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-iso Propyl-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-(pro p-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-triazole-1- 1)-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-iso Propyl-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-iso Propyl-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-iso Propyl-2-(4-methylpyrimidin-5-yl)isoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-iso Propyl-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-1 H -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-1 H -1,2,4-triazol-1-yl)-7-fluoro 4-iso Propyl-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-1 H -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-1 H -1,2,4-triazol-1-yl)-7-fluoro-4- Isopropyl-2-(5-methylpyrimidin-4-yl)isoquinolin-1(2H)-one;

2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H - 1,2,4-tri Azol-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-1 H -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-1 H -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-1 H -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-fluo Ro-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-iso Propyl-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-1 H -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-iso Propyl-2-(2-methoxy-5-methylpyridin-4-yl)isoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-7-fluoro-4- Isopropyl-2-((1 S ,2 R )-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-1 H -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-1 H -1,2,4-triazol-1-yl )-7-Fluoro-4-isopropylisoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-1 H -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-1 H -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-iso Propyl-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-iso Propyl-2-(5-methylisothiazol-4-yl)isoquinolin-1(2H)-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-1 H -1,2,4 -triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-1 H -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-1 H -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-1 H -1,2,4-tri Azol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5( 6H )-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- Isopropyl-6-( o -D ₃ -tolyl)-1,6-naphthyridin-5(6 H )-one;

and, optionally, one or more of pharmaceutically acceptable salts, isotopes, N-oxides, solvates, and stereoisomers thereof.

In an embodiment of the invention, the DHODH inhibitor is a compound 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)-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-triazole -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-triazole -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-triazole -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-triazole-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-triazole-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-triazole -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-Isopropylphthalazine-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-triazole -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-iso Propyl-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-triazole -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-triazole -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-triazole -1-yl)-7-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;

2-(5-chloro-3-methyl-1 H -pyrazol-4-yl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- (prop-1-en-2-yl)-6-( o -tolyl)-1,6-naphthyridin-5(6 H )-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- methyl-6-( o -tolyl)pyrido[2,3-d]pyridazin-5(6 H )-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-tri Azol-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-triazole -1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5(6H)-one;

(S)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluo Ro-6-( o -tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6 H )-one;

(R)-2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-3-fluo Ro-6-( o -tolyl)-8-(1,1,1-trifluoropropan-2-yl)-1,6-naphthyridin-5(6 H )-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- Isopropyl-6-( o -tolyl)-1,6-naphthyridin-5(6 H )-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-triazole-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-triazole -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-iso Propyl-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-triazole -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-iso Propyl-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-triazole -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-fluo Ro-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-fluo Ro-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-triazole -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-methoxyphenyl)-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-triazole- 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-triazole -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-triazole- 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-( 4-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-triazole-1- 1)-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-iso Propyl-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-(pro p-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-triazole-1- 1)-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-iso Propyl-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-iso Propyl-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-iso Propyl-2-(4-methylpyrimidin-5-yl)isoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-iso Propyl-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-1 H -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-1 H -1,2,4-triazol-1-yl)-7-fluoro 4-iso Propyl-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-1 H -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-1 H -1,2,4-triazol-1-yl)-7-fluoro-4- Isopropyl-2-(5-methylpyrimidin-4-yl)isoquinolin-1(2H)-one;

2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H - 1,2,4-tri Azol-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-1 H -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-1 H -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-1 H -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-fluo Ro-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-iso Propyl-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-1 H -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-iso Propyl-2-(2-methoxy-5-methylpyridin-4-yl)isoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -1,2,4-triazol-1-yl)-7-fluoro-4- Isopropyl-2-((1 S ,2 R )-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-1 H -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-1 H -1,2,4-triazol-1-yl )-7-Fluoro-4-isopropylisoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-iso Propyl-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-1 H -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-1 H -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-iso Propyl-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-iso Propyl-2-(5-methylisothiazol-4-yl)isoquinolin-1(2H)-one;

2-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-1 H -1,2,4 -triazol-1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -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-1 H -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-1 H -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-1 H -1,2,4-tri Azol-1-yl)-3-fluoro-8-isopropyl-1,6-naphthyridin-5( 6H )-one;

6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1 H -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-1 H -1,2,4-triazol-1-yl)-3-fluoro-8- Isopropyl-6-( o -D ₃ -tolyl)-1,6-naphthyridin-5(6 H )-one;

and, optionally, one or more of pharmaceutically acceptable salts, isotopes, N-oxides, solvates, and stereoisomers thereof; or

Compounds selected from:

2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole -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-triazole -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-triazole -1-yl)-6-methoxy-4-(prop-1-en-2-yl)phthalazin-1(2H)-one;

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

In an embodiment of the invention, the DHODH inhibitor is a compound of formula (Z) selected from the group consisting of:

2-(2-chloro-6-fluorophenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole -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-iso Propyl-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-triazole -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-triazole -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-iso Propyl-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-iso Propyl-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-triazole -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-iso Propyl-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-iso Propyl-2-(2-(trifluoromethyl)phenyl)isoquinolin-1(2H)-one; and

2-(2-(difluoromethyl)phenyl)-6-(4-ethyl-3-(hydroxymethyl)-5-oxo-4,5-dihydro-1H-1,2,4-triazole -1-yl)-7-fluoro-4-isopropylisoquinolin-1(2H)-one;

and, optionally, one or more of pharmaceutically acceptable salts, isotopes, N-oxides, solvates, and stereoisomers thereof.

In an embodiment of the invention, the DHODH inhibitor is a compound of Formula (Z) having Formula (Za), or a pharmaceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof:

[Chemical formula (Za)]

,

In the above equation

Y is CH or N;

R ¹ is 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;

R ² is ego;

R ³ is selected from the group consisting of H, halo, CH ₃ , and OCH ₃ ;

R ⁴ 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 ;

;

, and ; and

, and is selected from the group consisting of;

here,

Each R ^c is independently 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 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 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.

In an embodiment of the invention, the DHODH inhibitor is a compound of Formula (Z) having Formula (Zb), or a pharmaceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof:

[Chemical formula (Zb)]

,

In the above equation

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;

R ² ego;

R ³ is selected from the group consisting of H, halo and OCH ₃ ;

R ⁴ is

, and is selected from the group consisting of;

here,

R ^c is 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 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 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.

Hereinafter, exemplary compounds of formula (Z) useful in the methods of the present invention will be described with reference to the following exemplary synthetic schemes for their general preparation and the examples that follow. Those skilled in the art will recognize that in order to obtain the various compounds of the present specification, the starting materials can be appropriately selected so that the desired substituents may be protected or unprotected as needed through the reaction scheme to obtain the desired product. Alternatively, instead of the final desired substituent, it may be necessary or desirable to use an appropriate group that can be carried through the reaction scheme and substituted with the desired substituent as needed. Unless otherwise specified, variables are as defined above for Formula (Z). The reaction may 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 heating or microwave heating. The reaction can also be conducted in a closed pressure vessel above the normal reflux temperature of the solvent.

All abbreviations used in the general scheme and examples for Formula (Z) are as defined in Table 1a. Variables are as defined within the scope or as specifically defined in the general reaction equation.

[Table 1a]

Manufacturing example

From now on, exemplary compounds of formula (Z) useful in the methods of the present invention will be described with reference to the following exemplary synthetic schemes for their general preparation and the specific examples that follow.

Scheme 1

According to Scheme 1, the 1,2,4-triazol-5(4H)-one compound of formula (II) where PG is Bn is prepared from ethyl 2-(benzyloxy)acetate in three steps. 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 to 85° C. The hydrazide is prepared with an isocyanate of the formula R ^a -NCO, wherein R ^a is C _1-6 alkyl, in a suitable solvent such as water; The reaction provides the corresponding semicarbazide. Subsequently, the semicarbazide is cyclized with a suitable base such as NaOH in a suitable solvent such as water to provide a compound of formula (II) wherein PG is Bn.

Compounds of formula (II), wherein R ^a is C _1-6 haloalkyl or C _3-6 cycloalkyl, are suitably represented by formula R ^a -NCO, wherein R ^a is C _1-6 haloalkyl or C _3-6 cycloalkyl. Substituted compounds can be used and prepared as described above.

Protective group exchange of compounds of formula (II) where PG is Bn to compounds of formula (II) where PG is TBDPS is described in TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis," 3 ed., John Wiley & Sons. , 1999], using established methods. In the first step, deprotection of the benzyl group is achieved under hydrogenolysis conditions known to those skilled in the art to obtain the alcohol. For example, deprotection uses a palladium catalyst such as Pd/C; under H ₂ ; in a suitable solvent such as EtOH, MeOH, EtOAc, or mixtures thereof, preferably EtOH; With or without HCl; This is achieved over a period of 4 to 72 hours. In the second step, protection of the corresponding alcohol as a silyl ether using a suitable base such as tert- butyldiphenylsilyl chloride, imidazole, dimethylaminopyridine, pyridine, etc.; Among solvents such as DMF, DCM, etc.; At temperatures ranging from 0° C. to room temperature; This is achieved to obtain a compound of formula (II) wherein PG is TBDPS.

Scheme 2

According to Scheme 2, a compound of formula (XIV) where R ³ is H can be reacted with a halogenating reagent such as N-iodosuccinimide (NIS); In aprotic solvents such as acetonitrile; under heating conditions; Treatment gives the halogenated compound of formula (III) where HAL is iodide. Compounds of formula R ¹ -B(OH) ₂ are reacted with compounds of formula (III) under Suzuki coupling conditions known to those skilled in the art to provide compounds of formula (IV). For example, compounds of formula (III) wherein HAL is iodide can be prepared using commercially available or synthetically accessible boronic acids (or boronic acid esters) such as R ¹ -B(OH) ₂ wherein R ¹ has the formula: is optionally substituted C _2-6 alkenyl or aryl as defined for (Z); Palladium catalysts such as bis(triphenylphosphine)palladium(II) dichloride, tetrakis(triphenylphosphine)palladium, etc.; with a suitable base such as potassium phosphate, Cs ₂ CO ₃ , etc.; in a suitable solvent such as dioxane, water, ethanol, or mixtures thereof; The reaction provides a compound of formula (IV). Compounds of formula (IV) wherein R ³ is H, compounds of formula R ⁴ -B(OH) ₂ ; Reaction under copper (II) mediated Chan-Lam coupling conditions known to those skilled in the art provides compounds of formula (V) wherein HAL is ^bromide , For example, a compound of formula (IV) may be a compound of formula R ⁴ -B(OH) ₂ wherein R ⁴ is as defined for formula (Z); catalysts such as copper(II) acetate; bases such as pyridine, NEt ₃ , etc.; In suitable solvents such as DCM, ACN, dioxane, THF, etc.; React to obtain a compound of formula (V).

Scheme 3

According to Scheme 3, R ¹ is optionally substituted C _2-6 alkenyl, R ³ is H, R ⁴ is phenyl suitably substituted as described herein for Formula (Z), and HAL is A compound of formula (V) which is Br; Nucleophilic compounds of formula (II) wherein R ^a is C _1-6 alkyl, commercially available or synthetically accessible; with a suitably protected triazolone, for example where PG is selected from benzyl, 4-methoxy benzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES, or TIPS; In the presence of catalyst CuI and diamines such as trans-1,2-diaminocyclohexane, and bases such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc.; In suitable solvents such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc.; Ullmann-type aromatic amination reaction provides compounds of formula (VI) wherein X is CH and Y is CH.

A compound of formula (VI) wherein PG is Bn and R ¹ is C _2-6 alkenyl is reacted under Simmons-Smith cyclopropanation reaction conditions known to those skilled in the art to obtain a compound of formula (VI) wherein R ¹ is C _1-6 alkyl. Provided is a compound of formula (VI) wherein the C _3-6 cycloalkyl is substituted with . For example, R ¹ A compound of formula (VI), diiodomethane; diethyl zinc; Among suitable solvents such as toluene and the like; At temperatures ranging from 0° C. to room temperature; for a period of 3 to 26 hours; Reaction provides a compound of formula (VI) wherein R ¹ is cycloalkyl substituted with CH ₃ .

Subsequently, deprotection was performed using established methods, such as those described in TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis," 3 ed., John Wiley & Sons, 1999, to obtain Provides a compound of formula (Z) wherein is CH. For example, a compound of formula (VI) wherein R ³ is H and PG is TBDPS is deprotected using conditions known to those skilled in the art, preferably using 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 achieved using triethylamine trihydrogen fluoride (Et ₃ N·3HF).

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

X is CH; Y is CH; R ² , R ³ , R ⁴ are each defined as described herein for Formula (Z); Compounds of formula (Z) wherein R ¹ is C _2-6 alkenyl can be reacted using hydrogenation conditions known to those skilled in the art, for example under H ₂ with Pd/C or a Wilkinson catalyst [RhCl(PPh ₃ ) ₃ ]. using; In suitable solvents such as MeOH, THF, EtOAc, etc.; Reduction provides compounds of formula (Z) wherein R ¹ is C _2-6 alkyl.

Scheme 4

According to Scheme 4, compounds of formula (VII) can be reacted with α,β-unsaturated aldehydes such as 3-methyl-2-butenal, 3-methylpent-2-enal, etc.; TiCl ₄ ; and using bases such as triethylamine; Among aprotic solvents such as dichloromethane (DCM); Reductive amination gives the enamine intermediate, which is subsequently used with a reducing agent such as NaBH ₄ ; Reduction gives compounds of formula (VIII) wherein R ⁵ is C _1-4 alkyl and R ⁴ is defined as described herein for formula (Z). Compounds of formula (VIII) can be prepared using commercially available or synthetically accessible 4-bromo-2-iodobenzoyl chloride and bases such as triethylamine and 4-dimethylaminopyridine (DMAP); In anhydrous aprotic solvents such as dichloromethane (DCM); Coupling gives the compound of formula (IX). A compound of formula ( ^IX ) is treated with palladium(II) acetate, tetrabutylammonium bromide, and _potassium acetate under heated Heck reaction ^conditions to obtain CH and HAL is Br; and R ¹ is optionally substituted C _2-6 alkenyl, R ³ is H, X is CH ₂ and HAL is Br.

Scheme 5

According to Scheme 5, commercially available or synthetically accessible compounds of formula (X) wherein HAL is F, R ³ is F and R ⁵ is H or C _1-4 alkyl; Nucleophilic compounds of formula (II) wherein R ^a is C _1-6 alkyl, commercially available or synthetically accessible, such as PG is benzyl, 4-methoxy benzyl, or alkyl or aryl silanes such as TBDPS, TBS, TES, or a suitably protected triazolone selected from TIPS; in the presence of bases such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc.; Among suitable solvents such as DMSO, DMF, THF, ACN, etc.; The reaction gives a compound of formula (XI). In a preferred method, PG is Bn and R ^a is C _1-6 alkyl. The esters of formula (XI), when R ⁵ is C _1-4 alkyl, are hydrolyzed to their corresponding acids under acidic or basic conditions. For example, tert-butyl ester (R ⁵ is tert -Bu) is treated with TFA; Or alternatively, hydrolysis in an aqueous solvent using a base such as NaOH gives a compound of formula (XIa) wherein R ⁵ is H. Compounds of formula (XIa) are chlorinated using conditions known to those skilled in the art to provide acyl chlorides of formula (XII). For example, the compound of formula (XIa) is heated in SOCl ₂ ; Treated with oxalyl chloride in DCM.

Scheme 6

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; using a compound of formula (VIII) wherein R ⁵ is C _1-4 alkyl and a base such as a mixture of triethylamine (TEA) and 4-dimethylaminopyridine (DMAP); In anhydrous aprotic solvents such as dichloromethane (DCM); The reaction gives a compound of formula (XIII). Compounds _of ^Formula (VI) wherein Treatment under Heck reaction conditions using palladium (II) acetate, tetrabutylammonium bromide ^, _and potassium acetate gives a mixture of intramolecularly cyclized compounds, which are then separated to form The intermediate compound where ³ is H or F was isolated.

Scheme 7

According to Scheme 7, compounds of formula (XIV) wherein R ³ is H or F can be converted to compounds of formula (II); with a suitably protected triazolone, for example where PG is selected from benzyl, 4-methoxy benzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES, or TIPS; According to the previously described method, Ullmann-type aromatic amination reaction gives the compound of formula (XV). In a preferred method, PG is Bn and R ^a is C _1-6 alkyl. A compound of formula (XV) can be reacted with a halogenating reagent such as N-iodosuccinimide (NIS); In aprotic solvents such as acetonitrile; under heating conditions; Processing gives a halogenated compound of formula (XVI) wherein Y is CH and HAL is iodide.

Scheme 8

According to Scheme 8, compounds of formula (XVIIa) and formula (XVIIb) are prepared in two steps from 5-bromoisobenzofuran-1,3-dione. 5-bromoisobenzofuran-1,3-dione may be obtained commercially or synthetically accessible with suitably substituted alkyl Grignard reagents such as i-PrMgCl, EtMgBr, etc.; in the presence of CdCl ₂ ; In aprotic solvents such as THF and the like; reacted, followed by an alkylating agent of the formula R ⁵ -I wherein R ⁵ is C _1-4 alkyl (eg iodomethane or iodoethane); in the presence of bases such as K ₂ CO ₃ , Cs ₂ CO ₃ , etc.; In aprotic solvents such as DMF, DMSO, etc.; Processing gives a mixture of regio-isomeric esters of formula (XVIIa) and formula (XVIIb) wherein R ¹ is optionally substituted C _1-6 alkyl. In a similar manner, aryl Grignard reagents can be used to obtain compounds of formula (XVIIa) and formula (VXIIb), wherein R ¹ is a suitably substituted phenyl. The positional isomers of formula (XVIIa) and formula (XVIIb) are used directly without separation and converted to the corresponding phthalazinone (mixture). For example, a mixture of formula (XVIIa) and formula (XVIIb) with an excess of hydrazine; in a suitable solvent such as ethanol or methanol; At temperatures ranging from room temperature to 90°C; Treatment is for a period of approximately 6 to 20 hours. The desired phthalazinone compound of formula (V) can be easily separated from the remaining other regio-isomers by precipitation, crystallization, or purified by flash chromatography. Compounds of formula (V) are selected from formula (II) wherein R ^a is C _1-6 alkyl and PG is selected from benzyl, 4-methoxy benzyl, or an alkyl or aryl silane such as TBDPS, TBS, TES, or TIPS. a suitably protected triazolone; In the presence of catalyst CuI and diamines such as trans-1,2-diaminocyclohexane, and bases such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc.; In suitable solvents such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc.; Ullmann-type aromatic amination reaction gives compounds of formula (XVIII) where X is N.

A compound of formula (XVIII) wherein R ¹ is C _2-6 alkenyl is reacted under Simmons-Smith cyclopropanation reaction conditions known to those skilled in the art to form C _3-6 cycloalkyl wherein R ¹ is substituted by C _1-6 alkyl. Provided is a compound of formula (XVIII). For example, compounds of formula (XVIII) wherein R ¹ is C _2-6 alkenyl include diiodomethane; diethyl zinc; in a suitable solvent such as toluene; At temperatures ranging from 0° C. to room temperature; for a period of 24 to 26 hours; Reaction provides a compound of formula (XVIII) wherein R ¹ is cycloalkyl substituted with CH ₃ .

Scheme 9

According ^to Scheme ⁹ , compounds of _formula ⁽ By coupling with an aldehyde of the formula R ¹ -CHO that is C _1-6 alkyl; The corresponding ketone compound of formula (XIX) is obtained (a similar modification has been reported in Suchand et al, J. Org. Chem. 81, 6409-6423). For example, methyl 4-bromo-2-iodobenzoate with isobutyraldehyde; palladium catalysts such as Pd(OAc) ₂ ; Ag ₂ O; and an aqueous solution of tert-butyl hydroperoxide (TBHP); At a temperature of about 120°C; for a period of 10 to 14 hours; The reaction provides methyl 4-bromo-2-isobutyrylbenzoate. Ketone compounds of formula (XIX) can be selected from hydrazines R ⁴ -NHNH ₂ wherein R ⁴ is suitably substituted aryl, such as 2-chloro-6-fluorophenylhydrazine; Reaction gives a compound of formula (V) where X is N. Compounds of formula (V) can be subjected to a Ullmann-type aromatic amination reaction with a suitably protected triazolone (II) as described above to obtain formula (VI) wherein Y is CH and R ¹ is selected from C _1-6 alkyl. ) to obtain the compound.

Compounds of formula (VI ^{) wherein Y is CH, R 1 is} phenyl ^and It can be prepared in a similar manner using the previously described method, by coupling it with an aldehyde of -CHO.

Scheme 10

According to Scheme 10, a compound of the formula R ¹ -B(OH) ₂ ; Reaction with a compound of formula (XVI) under Suzuki coupling conditions known to those skilled in the art provides a compound of formula (XVIII) wherein For example, compounds of formula (XVI) wherein Y is CH and HAL is iodide can be prepared using commercially available or synthetically accessible boronic acids (or boronic acid esters) such as R ¹ -B(OH) ₂ where R ¹ is optionally substituted C _2-6 alkenyl, C _3-6 cycloalkyl or anyl as defined for formula (Z); palladium catalysts such as bis(triphenylphosphine)palladium(II) dichloride; with a suitable base such as potassium phosphate, Cs ₂ CO ₃ , etc.; in a suitable solvent such as dioxane, water, ethanol, or mixtures thereof; Reaction gives a compound of formula (XVIII) wherein X is CH. It was noted that during the coupling reaction as described above, loss of iodide during the reaction conditions afforded a compound of formula (XVIII) where X is CH and R ¹ is H. Compounds of formula (XVIII) wherein X is CH or N, compounds of formula R ⁴ -B(OH) ₂ ; By reacting under copper(II) mediated Chan-Lam coupling conditions known to those skilled in the art or previously described, X is CH or N, R ¹ is optionally substituted C _2-6 alkenyl, and R ³ is H or F and R ⁴ is phenyl suitably substituted as described herein for Formula (Z).

A compound of formula (XVIII) in which R ¹ is N(CH ₃ ) ₂ is prepared from a compound of formula (XVI) in which HAL is Br and PG is Bn. A compound of formula (XVI) is reacted with an amine such as NH(CH ₃ ) ₂ in water; At a temperature of about 110°C; Over a period of 96 hours; The reaction gives a compound of formula (XVIII) wherein R ¹ is N(CH ₃ ) ₂ and R ^a is C _1-6 alkyl. Compounds of formula (Z) wherein R ¹ is N(CH ₃ ) ₂ are prepared according to the method described above.

Compounds of formula (XVIII) wherein R ¹ is C _1-6 alkyl substituted by OH are prepared in two steps from compounds of formula (XVIII) wherein R ¹ is C _2-6 alkenyl and PG is Bn. In the first step, R ¹ is a compound of formula (XVIII) under oxidizing conditions such as NaIO ₄ , and K ₂ OsO ₄ .2H ₂ O or OsO ₄ ; In a suitable solvent such as THF/H ₂ O; At temperatures ranging from 0° C. to room temperature; for a period of about 48 to 72 hours; React to obtain a ketone intermediate compound. In a second step, the ketone intermediate compound is combined with a Grignard reagent such as methylmagnesium bromide; In a solvent such as diethyl ether; At temperatures ranging from 0° C. to room temperature; for a period of 3 to 30 hours; The reaction gives a compound of formula (XVIII) wherein R ¹ is C _1-6 alkyl substituted with OH.

Scheme 11

According to Scheme 11, 4,5-difluorophthalic anhydride can be reacted with a hydrazine compound of the formula R ⁴ -NHNH ₂ wherein R ⁴ is suitably substituted phenyl or heteroaryl, such as (2-chloro-6-fluorophenyl). hydrazine hydrochloride; Among acetic acid; At a temperature of about 125°C; After reacting for a period of about 1.5 hours, a compound of formula (XX) wherein R ³ is F is obtained. Under basic conditions such as sodium ethoxide or sodium methoxide, depending on the rearrangement of the compound of formula (XX); In aprotic solvents such as ethanol, methanol, etc.; At room temperature; This is carried out over a period of about 1.5 hours to obtain the ring-expanding compound of formula (XXI). _{Trifluoromethanesulfonyl} ( Derivatization of compounds of formula (XXI) with sulfonate-based leaving groups such as triflate is achieved to obtain compounds of formula (XXII). Milder triflatifying agents such as N-phenylbis(trifluoromethanesulfonimide) (Tf ₂ NPh), bases such as TEA, DIEA, etc. may be used in suitable solvents such as DCM.

Scheme 12

인 화학식 (V)의 화합물을, Pd/C 등과 같은 촉매를 사용하는 수소화 조건 하에서, EtOAc 등과 같은 적합한 용매 중에; 실온에서 수소 가스(20 내지 45 psi)의 분위기 하에; 4 내지 24시간의 기간 동안; 반응시켜 R¹이

인 화학식 (V)의 화합물을 수득한다. 앞서 기재된 조건을 사용하여 화학식 (V)의 화합물과 화학식 (II)의 적합하게 보호된 트라이아졸론의 반응을 수행하여 화학식 (VI) 및 화학식 (VIa)의 화합물의 혼합물을 얻으며, 이것은 보호기의 탈보호 전 또는 후에 분리될 수 있다.According to Scheme 12, a compound of the formula R ¹ -B(OH) ₂ ; Reaction of a compound of formula (XXII) under the Suzuki coupling conditions previously described provides a compound of formula (V) wherein For example, compounds of formula (XXII) can be prepared using commercially available or synthetically accessible boronic acids (or boronic acid esters), such as R ¹ having C _2-6 as defined herein for formula (Z). R ¹ -B(OH) ₂ which is alkenyl or C _2-6 haloalkenyl; Palladium catalysts such as 1,1'-bis(diphenylphosphine)ferrocene-palladium(II) dichloride or bis(triphenylphosphine)palladium(II) dichloride; with a suitable base such as potassium phosphate, Cs ₂ CO _{3 ,} K ₂ CO ₃ , etc.; in a suitable solvent such as dioxane, water, ethanol, or mixtures thereof; The reaction provides a compound of formula (V). Selective hydrogenation of a compound of formula (V) wherein R ¹ is C _2-6 alkyl or C _2-6 haloalkyl of a compound of formula (V) wherein R ¹ is C _2-6 alkenyl or C _2-6 haloalkenyl It is easily manufactured by. For example, R ¹

A compound of formula (V) is reacted in a suitable solvent such as EtOAc under hydrogenation conditions using a catalyst such as Pd/C; under an atmosphere of hydrogen gas (20 to 45 psi) at room temperature; for a period of 4 to 24 hours; After reacting, R ¹ becomes

A compound of formula (V) is obtained. The reaction of a compound of formula (V) with a suitably protected triazolone of formula (II) is carried out using the conditions previously described to give a mixture of compounds of formula (VI) and formula (VIa), which leads to the desorption of the protecting group. Can be separated before or after protection.

Scheme 13

According to Scheme 13, fluoro compounds of formula (XXIII) wherein suitably substituted commercially available or synthetically accessible R ^c and R ^d are as defined for formula (Z) are reacted to give compounds of formula (XVIII). Achieve N-arylation of. 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 Compounds of (XVIII) can be combined with fluoro compounds of formula (XXIII), which are commercially available or accessible by synthesis, under nucleophilic displacement reaction conditions; in the presence of bases such as K ₂ CO ₃ , Cs ₂ CO ₃ , etc.; Among aprotic solvents such as DMF, DMSO, etc.; at temperatures ranging from 65 to 100° C.; The reaction provides a compound of formula (XXIV).

Reduction of compounds of formula (XXIV) using zinc or iron and NH ₄ in a mixed solvent of methanol and water; This can be achieved to provide an amino compound of formula (XXV).

Compounds of formula (XXV) with NaNO ₂ ; In acidic aqueous solutions or other nitrite reagents; In organic solvents such as EtOH; Diazotization at a temperature of 0° C.; Subsequently reducing the diazo group with zinc at a temperature ranging from 0 to 85° C.; or by treatment with H ₃ PO ₂ ; Provided are compounds of formula (XXVI) wherein R ^c and R ^d are defined as described herein for formula (Z).

Scheme 14

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

R ¹ An isocoumarin compound of formula (XXX) is prepared from a compound of formula (XIa) and methylbuta-1,2-dien-1-yl acetate. Methylbuta-1,2-dien-1-yl acetate 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 ; Among suitable solvents such as DCM, etc.; The reaction provides 2-methylbut-3-yn-2-yl acetate. 2-methylbut-3-yn-2-yl acetate is mixed with a catalytic amount of a Lewis acid such as AgBF ₄ , AgClO ₄ , PtCl ₂ , etc.; The reaction provides 3-methylbuta-1,2-dien-1-yl acetate. 3-Methylbuta-1,2-dien-1-yl acetate is reacted with a compound of formula (XIa) in which R ⁵ is H and Catacxium A in a suitable solvent such as DMF under Heck reaction conditions as previously described. using phase transfer reagents such as Pd G2, and Cy ₂ NMe palladium(II) acetate, tetrabutylammonium bromide, and bases such as potassium acetate; at a temperature of 70°C to 90°C; for a period of 10 to 16 hours; Coupling using intermolecular cyclization such that Y is CH and R ¹ is Provided is an isocoumarin compound of formula (XXX).

Y is CH, and R ¹ is A compound of formula (XXX) is reacted in a suitable solvent such as THF under hydrogenation conditions using a Wilkinson catalyst [RhCl(PPh ₃ ) ₃ ] or the like; Selective reduction at room temperature provides isocoumarin compounds of formula (XXX) wherein R ¹ is isopropyl.

Scheme 15

According to Scheme 15, 2-butanone is converted to ethyl 3-methylpent-2-enoate using Wittig reaction conditions known to those skilled in the art. For example, 2-butanone can be combined with triphenyl phosphonium ylide, such as (carbethoxymethylene) triphenylphosphorane, with or using additives such as benzoic acid, LiCl, and sodium dodecyl sulfate (SDS), etc. Instead, the reaction is carried out in a suitable solvent such as toluene at a temperature ranging from room temperature to the reflux temperature of the solvent for a period of 12 to 24 hours. Ethyl 3-methylpent-2-enoate is converted to 3-methylpent-2-ene-1- in a suitable solvent such as toluene using a suitable reducing agent such as DIBAL-H at a temperature ranging from -78° C. to room temperature. Reduce to ol. 3-Methylpent-2-en-1-ol was reacted under oxidizing conditions known to those skilled in the art, such as DMP (Dess-Martin periodinane), SO ₃ -pyridine, Swern conditions [(COCl) ₂ , DMSO, Et ₃ N], PCC, etc. are oxidized to 3-methylpent-2-enal in a solvent such as EtOAc, DMSO, DCM, etc. at a temperature ranging from about -78°C to room temperature (about 23°C). In a preferred method, 3-methylpent-2-en-1-ol is oxidized to 3-methylpent-2-enal using Dess-Martin periodinane in DCM at 25° C. for a period of 1 to 4 hours. .

Scheme 16

According to Scheme 16, isocoumarin compounds of compounds of formula (XXX) where Y is CH can be prepared commercially or synthetically accessible where R ⁴ has the formula R ⁴ -NH as defined herein for formula (Z). Compound of ₂ ; Lewis acids such as AlMe ₃ and AlCl ₃ ; in a suitable aprotic solvent such as DCM, toluene, etc.; Reaction provides compounds of formula (XXXI), wherein Y is CH and R ¹ , R ³ , R ⁴ and R ^a are as defined herein for formula (Z).

Scheme 17

Isocoumarin compounds of formula (XXX) can be prepared according to Scheme 17. ^4,5 -difluoro-2 ^- iodobenzoyl chloride, from compounds of formula ( Using conditions known to the general public, in the presence of a catalytic amount of DMF, in a suitable aprotic non-polar solvent such as dichloromethane (DCM), tetrahydrofuran (THF), acetonitrile (ACN), toluene, etc., at 0° C. Manufactured at a temperature ranging from room temperature,

It forms 4,5-difluoro-2-iodobenzoyl chloride. 4,5-difluoro-2-iodobenzoyl chloride with 2-methylbut-3-yn-2-ol, which is commercially available or synthetically accessible, in the presence of bases such as triethylamine and DMAP; Among suitable solvents such as DCM, etc.; The reaction can be performed to obtain an ester compound of formula (XXXIII). Compounds of formula (XXXIII) can be reacted with compounds of formula (II) using methods as previously described to obtain compounds of formula (XXXIV). A compound of formula (XXXIV) is reacted with a catalytic amount of a Lewis acid such as AgClO ₄ , PtCl ₂ , etc.; Processing can yield rearranged compounds of formula (XXXV). Compounds of formula (XXXV), wherein R ³ is H or F, can be reacted with a suitable solvent such as DMF, under Heck reaction conditions, using a phase transfer reagent such as palladium(II) acetate, tetrabutylammonium bromide, and a base such as potassium acetate. Among solvents; at a temperature of 70°C to 90°C; for a period of 1 to 3 hours; Intramolecular cyclization can provide isocoumarin compounds of formula (XXX) wherein Y is CH.

Scheme 18

According to Scheme 18, compounds of formula (Xa) wherein HAL is Cl, R ⁵ is CH(CH ₃ ) ₂ , and R ³ is F are commercially available or disclosed in US Patent Application Publication No. US2016-0176869 (Chen, et al. ) is accessible by synthesis according to the method as described. Compounds of formula (Xa), commercially available or synthetically accessible, include nucleophilic compounds of formula (II) wherein PG is benzyl and R ^c is C _1-6 alkyl; In the presence of bases such as K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc.; In suitable solvents such as dimethyl sulfoxide (DMSO), DMF, THF, ACN, etc.; The reaction gives a compound of formula (XXXIX) where Y is N. Compounds of formula (XXXIX) or formula (XI) wherein R ³ is F and R ⁵ is C _1-4 alkyl include commercially available 1-ethoxyethene-2-boronic acid pinacol ester; Palladium catalysts such as bis(triphenylphosphine)palladium(II) dichloride, 1,1'-bis(diphenylphosphine)ferrocene-palladium(II) dichloride, etc.; with a suitable base such as Cs ₂ CO ₃ ; in a suitable solvent such as dioxane, water, ethanol, or mixtures thereof; using conventional heating or microwave heating; Reaction gives compounds of formula (XL) wherein Y is N or CH.

Scheme 19

According to Scheme 19, R ⁴ represents a compound of formula R ⁴ -NH ₂ as defined herein for formula (Z); trimethyl aluminum; reacting in a suitable solvent such as dichloromethane, toluene, or mixtures thereof; Combining the resulting solution with a compound of formula (XL) wherein Y is CH or N; Provided is a compound of formula (XLI). Compounds of formula (XLI), wherein Y is CH or N, are treated with acetic acid or trifluoroacetic acid under heating conditions of 50° C. to 90° C. to provide compounds of formula (XLII). Halogenation of a compound of formula (XLII) using N -bromosuccinimide in anhydrous dimethylformamide at room temperature provides a compound of formula (XVI) wherein HAL is Br. Compounds of the formula R ¹ -B(OH) ₂ ; By reacting with a compound of formula (XVI) under Suzuki coupling conditions known to those skilled in the art or as previously described, R ¹ is C _2-6 alkenyl, C ₂ , optionally substituted as described herein for formula (Z). _-6 haloalkenyl, or aryl. A compound of formula (VI) wherein R ¹ is optionally substituted C _2-6 alkenyl or C _2-6 haloalkenyl is reacted under hydrogenation conditions using a Wilkinson catalyst ((PPh ₃ ) ₃ RhCl) to give R ¹ C _2-6 alkyl or C _2-6 haloalkyl.

Scheme 20

According to Scheme 20, 3-methylbutanal is reacted with a compound of formula (XXXIX) wherein Y is N and R ⁵ is CH(CH ₃ ) ₂ and a palladium catalyst such as allylpalladium(II) chloride dimer; Ligands such as 1,1'-bis(diphenylphosphino)ferrocene (dppf); using a suitable base such as Cs ₂ CO ₃ ; In the presence of a moisture scavenger such as molecular sieve (4A); In a suitable solvent such as dioxane; Reaction provides compounds of formula (XXX) wherein R ¹ is isopropyl. a compound of formula R ⁴ -NH ₂ wherein R ⁴ is as defined herein for formula (Z); trimethyl aluminum; reacting in a suitable solvent such as dichloromethane, toluene, or mixtures thereof; The resulting solution is combined with a compound of formula (XXX), followed by subsequent treatment with acetic acid under a heating temperature of 80° C. to 100° C. for a period ranging from 5 ^to 24 hours, wherein isopropyl and R ³ is F.

Scheme 21

According to Scheme 21, a compound of Formula (XXXIX) or Formula (XI) wherein R ³ is F and R ⁵ is C _1-4 alkyl; commercially available vinylboronic acid pinacol ester; palladium catalysts such as bis(triphenylphosphine)palladium(II) dichloride, or 1,1'-bis(diphenylphosphine)ferrocene-palladium(II) dichloride dichloromethane complex; with a suitable base such as Cs ₂ CO ₃ ; in a suitable solvent such as dioxane, water, ethanol, or mixtures thereof; Reaction provides compounds of formula (XLIII) wherein 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 potassium (VI) osmate dihydrate/sodium periodate, or ozonolysis. Compounds of formula (XLIV) are commercially available or synthetically accessible using suitably substituted alkyl Grignard reagents such as i-PrMgCl and the like; In aprotic solvents such as THF and the like; After the reaction, subsequent treatment is performed using an oxidizing reagent such as Dess-Martin reagent or Sweir oxidation conditions to obtain a ketone compound of formula (XLV).

Compounds of formula (XLV) are prepared in two steps from compounds of formula (XXXIX). compounds of formula (XXXIX) wherein R ³ is F and R ⁵ is C _1-4 alkyl; commercially available tributyl(1-ethoxyvinyl)tin; Palladium catalysts such as bis(triphenylphosphine)palladium(II) dichloride, 1,1'-bis(diphenylphosphine)ferrocene-palladium(II) dichloride, etc.; React in a suitable solvent such as dioxane, water, ethanol or mixtures thereof. Subsequently, acidic hydrolysis is performed using conditions such as treatment with aqueous HCl solution at room temperature to obtain compounds of formula (XLV) wherein X is N, Y is N or CH and R ¹ is methyl.

Commercially available or synthetically available R ⁴ is hydrazine R ⁴ -NHNH ₂ as defined herein for formula (Z), such as 2-chloro-6-fluorophenylhydrazine, o -tolylhydrazine; Compounds of formula (XLV); In the presence of a base such as potassium carbonate, etc.; Under heating conditions such as 70°C to 120°C; in a suitable solvent such as toluene, or mixtures thereof; Condensation gives compounds of formula (VI) wherein X is N, Y is CH or N and R ⁴ is as defined herein for formula (Z).

Scheme 22

According to Scheme 22, methyl 2-bromo-4,5-difluorobenzoate is combined with a suitably protected triazolone compound of formula (II); in the presence of bases such as K ₃ PO ₄ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaHCO ₃ , triethylamine, etc.; In suitable solvents such as 1,4-dioxane, DMSO, DMF, THF, ACN, etc.; Reaction gives a 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). 3-methylbutanal is reacted with a compound of formula XLVI and a palladium catalyst such as allylpalladium(II) chloride dimer; Ligands such as 1,1'-bis(diphenylphosphino)ferrocene (dppf); with a suitable base such as Cs ₂ CO ₃ ; In the absence of a moisture scavenger such as molecular sieve (4A); In a suitable solvent such as dioxane; The reaction provides a compound of formula (XLVII). a compound of formula R ⁴ -NH ₂ wherein R ⁴ is as defined herein for formula (Z); trimethyl aluminum; reacting in a suitable solvent such as dichloromethane, dichloroethane, toluene, or mixtures thereof; The resulting solution is combined with a compound of formula (XLVII), followed by subsequent treatment with acetic acid under a heating temperature of 80° C. to 100° C. for a period ranging from 5 ^to 24 hours, so that isopropyl and R ³ is F. In certain cases, compounds of the formula R ⁴ -NH ₂ , such as o-toluidine and the like; A compound of formula (XLVII) in acetic acid at a heating temperature of 80° C. to 100° C. for a period ranging from 10 to 24 hours; Direct condensation provides a compound of formula (VI) wherein X is CH, Y is N, R ¹ is isopropyl and R ³ is F.

Scheme 23

According to Scheme 23, a compound of formula (XXXIX) wherein R ⁵ is C _1-4 alkyl, Y is N and R ³ is F can be prepared by reacting a silyl protected alkyne such as trimethylsilylacetylene, palladium(II)bis( Triphenylphosphine) Palladium catalysts such as triphenylphosphine dichloride; copper catalysts such as copper iodide; The Sonogashira coupling reaction is carried out in a suitable solvent such as ACN, toluene, etc., using a suitable base such as triethylamine. in a suitable solvent such as THF using TBAF; The deprotection reaction is performed at room temperature to obtain the compound of formula (XLVIII). Compounds of formula (XLIX) are obtained using a gold catalyst, preferably AuCl ₃ , in a suitable solvent mixture such as MeCN. A similar transformation by AuCl ₃ -catalyzed cyclization is described in Marchal, E. et al in Tetrahedron 2007, 63, 9979-9990. a compound of formula R ⁴ -NH ₂ wherein R ⁴ is as defined herein for formula (Z); trimethylaluminum; reacting in a suitable solvent such as dichloromethane, dichloroethane, toluene, or mixtures thereof; The resulting solution is combined with the compound of formula (XLIX), followed by subsequent treatment with acetic acid under a heating temperature of 80° C. to 100° C. for a period ranging from 5 to 24 hours to provide the compound of formula (L). NBS is used at room temperature in a suitable solvent, preferably DMF, followed by a palladium catalyst, preferably palladium(II)bis(triphenylphosphine)dichloride, Cs ₂ CO ₃ , using conditions known to those skilled in the art. using bases such as Na ₂ CO ₃ ; In a solvent mixture consisting of 1,4-dioxane and water; Cross-coupling at a temperature of 100° C. provides compounds of formula (VI) wherein X is CH, Y is N, R ³ is F, and R ¹ , R ^a and PG are defined as previously described.

Scheme 24

According to Scheme 24, the compound of formula (VI), wherein PG is Bn, is reacted using conditions known to those skilled in the art, preferably in TFA, at a temperature of about 60 to 90° C. in a sealed tube; or deprotection in a suitable solvent such as DCM using BCl ₃ at a temperature of -78° C.; or treatment with hydrogen gas in the presence of a catalyst such as palladium on carbon (Pd/C) to obtain compounds of formula (Z).

In a similar manner, N-arylation and in situ TBDPS deprotection of compounds of formula (XVIII) wherein R ¹ is I, PG is TBDPS and Compound Z) is obtained.

A compound of formula (Z) wherein R ³ is F is reacted by a nucleophilic aromatic substitution reaction to provide a compound of formula (Z) wherein R ³ is OCH ₃ . For example, a compound of formula (Z) wherein R ³ is F may be reacted with a suitable base such as NaOH; in a suitable solvent such as MeOH; Reaction gives a compound of formula (Z) wherein Y is CH and R ³ is OCH ₃ .

Compounds of formula (Z) can 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 give the corresponding salt forms. do. Alternatively, trifluoroacetic acid or formic acid salts are obtained as a result of reversed phase HPLC purification conditions. The crystalline form of the pharmaceutically acceptable salt of a compound of formula (Z) can be prepared by recrystallization from a polar solvent (including mixtures of polar solvents and aqueous mixtures of polar solvents) or a non-polar solvent (including mixtures of non-polar solvents) to form a crystalline form. It can be obtained in the form

If the compounds according to the invention have at least one chiral center, they may therefore exist as enantiomers. If a compound has two or more chiral centers, it may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are included within the scope of this invention.

Compounds prepared according to the above-described schemes can be obtained as a single form, such as a single enantiomer, by conformation-specific synthesis or resolution. Compounds prepared according to the above schemes can alternatively be obtained as mixtures in various forms, such as racemic (1:1) or non-racemic (not 1:1) mixtures. When racemic and non-racemic mixtures of enantiomers are obtained, single enantiomers can be separated by conventional separation methods known to those skilled in the art, such as chiral chromatography, recrystallization, formation of diastereomeric salts, derivatization into diastereomeric adducts, It can be isolated using biotransformation or enzymatic transformation. When regioisomeric or diastereomeric mixtures are obtained, if applicable, single isomers may be separated using conventional methods, such as chromatography or crystallization.

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

Example

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 stated, the reaction mixture was magnetically stirred at room temperature (rt) under a nitrogen atmosphere. When "drying" a solution, it is usually dried with a desiccant, such as Na ₂ SO ₄ or MgSO ₄ . When "concentrating" mixtures, solutions and extracts, they are typically concentrated on a rotary evaporator under reduced pressure.

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

Preparative reversed-phase high-performance liquid chromatography (RP HPLC) was performed in either:

Method A. Phenomenex Synergi C18 (10 μm, 150 % FA) and then Gilson GX-281 semi-prep-HPLC with the mobile phase held at 100% ACN for 2 min. or

Method B. Phenomenex Synergi C18 (10 μm, 150 × 25 mm) or Boston Green ODS C18 (5 μm, 150 × 30 mm) and 5 to 99% ACN (0.1% ACN) in water over 10 min at a flow rate of 25 mL/min. % TFA) and then Gilson GX-281 semi-prep-HPLC with the mobile phase held at 100% ACN for 2 min. or

Method C. Phenomenex Synergi C18 (10 μm, 150 % HCl) and then a Gilson GX-281 semi-prep-HPLC with the mobile phase held at 100% ACN for 2 min. or

Method D. Phenomenex Gemini C18 (10 μm, 150 × 25 mm), AD (10 μm, 250 mm × 30 mm), or Waters Gilson GX-281 semi-prep-HPLC with a mobile phase of 0 to 99% ACN in water (with 0.05% (v/v) ammonia hydroxide) over a period of 2 min followed by a 2 min hold at 100% ACN. or

Method E. Phenomenex Gemini C18 (10 μm, 150 × 25 mm) or Waters Gilson GX-281 semi-prep-HPLC with mobile phase held in mM NH ₄ HCO ₃ ) followed by 100% ACN for 2 min.

Preparative supercritical fluid high-performance liquid chromatography (SFC) was performed on a Thar 80 preparative-SFC system, or a Waters 80Q preparative-SFC system from Waters. In order to maintain CO ₂ in SF conditions, ABPR was set to 100 bar, and the flow rate was in the range of 50 g/min to 70 g/min, which can be confirmed depending on the compound characteristics. The column temperature was ambient.

Unless otherwise indicated, mass spectra (MS) were obtained using electrospray ionization (ESI) in positive ion mode on a SHIMADZU LCMS-2020 MSD or Agilent 1200/G6110A MSD. The calculated mass (calcd.) corresponds to the exact mass.

Nuclear magnetic resonance (NMR) spectra were obtained on a Bruker model AVIII 400 spectrometer. The definitions for multiplicity are: s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet, ddd = doublet, td = triplet, dt = doublet. , spt = septet, quin = quintet, m = multiplet, br = broad. It will be appreciated that for compounds containing exchangeable protons, the protons may be visible or invisible in the NMR spectrum depending on the concentration of the compound in solution and the choice of solvent used to run the NMR spectrum.

Chemical names were generated using ChemDraw Ultra 12.0, ChemDraw Ultra 14.0 (CambridgeSoft Corp., Cambridge, MA, USA), or ACD/Name version 10.01 (Advanced Chemistry).

Compounds designated R* or S* are enantiomeric pure compounds with no absolute configuration 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 hours. 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 hours. To the mixture was added H ₂ O (20 mL) and an aqueous solution (120 mL) of NaOH (57.7 g, 1.44 mol). The mixture was stirred at 95°C for 12 hours. The reaction mixture was cooled to room temperature and then quenched with HCl (12 M) at 0° C. and adjusted to “pH” 6. The solid was filtered and dried under reduced pressure to obtain the title compound (61 g, 91% yield) as a white solid. 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.

5-[(benzyloxy)methyl]-4-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (8 g, 34.3 mmol, 1.0 eq) in methanol (200 mL) .) Pd/C (2 g) was added to the solution. The resulting mixture was kept under hydrogen and stirred at room temperature for 6 hours. The resulting mixture was then filtered and the filtrate was concentrated to yield the crude product 4-ethyl-5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazole as a white solid. -3-one (4.3 g, 88% yield) was obtained. ^1H 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.

4-ethyl-5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazol-3-one (3 g, 21 mmol, 1.0 eq.) in DCM (30 mL) To the solution, tert -butylchlorodiphenylsilane (6.5 mL, 25 mmol, 1.2 eq.) and pyridine (1.86 mL, 23 mmol, 1.1 eq.) were added. The resulting mixture was stirred at room temperature overnight. 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 as a white solid. ,4-dihydro-3H-1,2,4-triazol-3-one (4.9 g, 61% yield) was obtained. LCMS (ES-API): mass calculated for C ₂₁ H ₂₇ N ₃ O ₂ Si, 381.2; m/z actual value, 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 ㎐, 2H), 1.33 (t, J = 7.34 ㎐, 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), followed by the addition of di-tert-butyl dicarbonate (4.6 g, 21 mmol). Then, DMAP (645 mg, 5.3 mmol) was added. The reaction mixture was stirred at 50° C. under nitrogen overnight and then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was diluted with EtOAc and then 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 (2.9 g, yield: 79%) as a yellow oil. 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.

tert-Butyl 4,5-difluoro-2-iodobenzoate (3.2 g, 9.4 mmol), 3-((benzyloxy)methyl)-4-ethyl-1H-1, in anhydrous DMF (30 mL). A mixture of 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) was stirred under nitrogen at 75° C. for 1 h, 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 (5 g, yield: 96%) as a colorless amorphous solid. ESI-MS: mass calculated for C ₂₃ H ₂₅ FIN ₃ O ₄ , 553.1; m/z actual value, 554.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.16 (d, J = 7.1 Hz, 1 H), 7.62 (d, J = 10.8 Hz, 1 H), 7.29 - 7.45 (m, 5 H), 4.61 (s , 2 H), 4.50 (s, 2 H), 3.84 (q, J = 7.2 Hz, 2 H), 1.63 (s, 9 H), 1.35 (t, J = 7.2 Hz, 3 H) ppm; ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -119.09 (dd, J = 10.6, 7.0 Hz, 1 F) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluo Ro-2-iodobenzoic acid .

tert -Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1- in DCM (50 mL) TFA (10 mL) was slowly added to a solution of 1)-5-fluoro-2-iodobenzoate (5 g, 9 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum. The resulting residue was triturated with petroleum ether for 30 minutes at room temperature. The mixture was filtered and the solid was rinsed with petroleum ether. The precipitate was collected and dried in vacuo to give the title compound (4.1 g, yield: 91%) as a white solid. ESI-MS: mass calculated for C ₁₉ H ₁₇ FIN ₃ O ₄ , 497.0; m/z actual value, 498.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.16 (d, J = 7.3 Hz, 1 H), 7.78 (d, J = 11.0 Hz, 1 H), 7.28 - 7.43 (m, 5 H), 4.60 (s, 2 H), 4.57 (s, 2 H), 3.74 (q, J = 7.2 Hz, 2 H), 1.23 (t, J = 7.2 Hz, 3 H) ppm; ¹⁹ F NMR (376 MHz, DMSO- d ₆ ) δ -119.91 (s, 1 F) 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.

3-Methylbuta-1,2-dien-1-yl acetate (Intermediate 12, 280 mg, 2.2 mmol), 4-(3-((benzyloxy)methyl)-4-ethyl- in DMF (7 mL) 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), Catacxium A Pd G2 (74.2 mg, 0.11 mmol) was added under nitrogen. The reaction mixture was stirred overnight at 90° C. under nitrogen. 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 (240 mg, yield: 25%) as a yellow solid. ESI-MS: mass calculated for C ₂₄ H _{2 2} FN ₃ O ₄ , 435.2; m/z 436.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.15 (d, J = 10.5 Hz, 1 H), 7.86 (d, J = 6.8 Hz, 1 H), 7.30 - 7.45 (m, 5 H), 7.19 (s , 1 H), 5.37 - 5.39 (m, 1 H), 5.18 (s, 1 H), 4.62 (s, 2 H), 4.53 (s, 2 H), 3.87 (q, J = 7.1 Hz, 2 H ), 2.11 (s, 3 H), 1.37 (t, J = 7.2 Hz, 3 H) ppm.

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

Method I:

Step A. 3-Methylbut-2-en-1-yl 4,5-difluoro-2-iodobenzoate.

1-Bromo-3-methyl in 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 dry DMF (20 mL). -2-Butene (1.5 g, 9.9 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours. 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: 10-20% EtOAc in heptane) to give the desired product (1.6 g, yield: 92%) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.80 (dd, J =7.58, 9.54 ㎐, 1H), 7.73 (dd, J = 7.83, 10.76 ㎐, 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.

3-Methylbut-2-en-1-yl 4,5-difluoro-2-iodobenzoate (1.6 g, 4.5 mmol), 3-((benzyloxy)methyl) in anhydrous DMF (25 mL) To a mixture of -4-ethyl-1H-1,2,4-triazol-5(4H)-one (Intermediate 1, 2.1 g, 9.1 mmol) was added Cs ₂ CO ₃ (2.9 g, 9.1 mmol). The reaction mixture was heated at 85° C. under nitrogen for 1 hour 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 (2.4 g, yield: 93%) as a white solid. LCMS (ES-API): mass calculated for C ₂₄ H ₂₅ FIN ₃ O ₄ , 565.1; m/z actual value, 566.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) 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-di Hydro-3H-1,2,4-triazol-3-one .

3-Methylbut-2-en-1-yl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1 in toluene (200 mL) (tBu ₃ P)PdG ₂ (351 mg, 0.69 mmol, 0.1 eq) and N-cyclohexyl-N-methyl-cyclohexanamine (1.60 mL, 7.54 mmol, 1.1 eq) were added separately. The reaction mixture was degassed with nitrogen three times and then heated at 80° C. for 18 hours under a nitrogen atmosphere. LCMS analysis showed that approximately 18% of the 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 eq) and tBu ₃ PPdG ₂ (176 mg, 0.34 mmol, 0.05 eq) were added. . The reaction mixture was degassed with nitrogen and then heated at 80° C. for 16 hours under a nitrogen atmosphere. The mixture was concentrated under reduced pressure, 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 x 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 (1.1 g, yield: 35%) as a yellow oil. ESI-MS: mass calculated for C ₂₄ H ₂₄ FN ₃ O ₄ , 437.2; m/z actual value, 438.5 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) 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:

5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-1-oxo-4-(prop-1-en-2-yl)-1H in THF (100 mL) at room temperature. -Isochromen-6-yl)-2,4-dihydro-3H-1,2,4-triazol-3-one (Intermediate 3, 5.9 g, 13.5 mmol) was added to a mixture of Wilkinson catalyst [RhCl(PPh) ₃ ) ₃ ] (3.8 g, 4.1 mmol) was added. The mixture was degassed and purged with hydrogen gas. The reaction mixture was stirred at room temperature under a hydrogen atmosphere (15 Psi) for 12 hours. The mixture was concentrated. The residue was purified by silica column chromatography (eluent: 0-25% EtOAc in petroleum ether) to give the title compound (1.5 g, yield: 77%) as a yellow solid. ESI-MS: mass calculated for C ₂₄ H ₂₄ FN ₃ O ₄ , 437.2; m/z actual value 438.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.10 (d, J = 10.5 Hz, 1 H), 8.01 (d, J = 7.0 Hz, 1 H), 7.46 (s, 1 H), 7.26 - 7.42 (m, 5 H), 4.61 (s, 2 H), 4.59 (s, 2 H), 3.77 (q, J = 7.3 Hz, 2 H), 3.08 (dt, J = 13.4, 6.8 Hz, 1 H) , 1.22 - 1.28 (m, 9 H) ppm; ¹⁹ F NMR (376 MHz, DMSO- d ₆ ) δ -118.29 (br s, 1 F) ppm.

Intermediate 5: 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluo Rho-2-iodobenzoyl 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), followed by the addition of di-tert-butyl dicarbonate (4.6 g, 21 mmol). Then, DMAP (645 mg, 5.3 mmol) was added. The reaction mixture was stirred at 50° C. under nitrogen overnight and then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was diluted with EtOAc and then 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 (2.9 g, yield: 79%) as a yellow oil. 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.

tert-Butyl 4,5-difluoro-2-iodobenzoate (3.2 g, 9.4 mmol), 3-((benzyloxy)methyl)-4-ethyl-1H-1, in anhydrous DMF (30 mL). A mixture of 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) was stirred under nitrogen at 75° C. for 1 h, 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 (5 g, yield: 96%) as a colorless amorphous solid. ESI-MS: mass calculated for C ₂₃ H ₂₅ FIN ₃ O ₄ , 553.1; m/z actual value, 554.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.16 (d, J = 7.1 Hz, 1 H), 7.62 (d, J = 10.8 Hz, 1 H), 7.29 - 7.45 (m, 5 H), 4.61 (s , 2 H), 4.50 (s, 2 H), 3.84 (q, J = 7.2 Hz, 2 H), 1.63 (s, 9 H), 1.35 (t, J = 7.2 Hz, 3 H) ppm; ¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -119.09 (dd, J = 10.6, 7.0 Hz, 1 F) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluo Ro-2-iodobenzoic acid .

tert -Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1- in DCM (50 mL) TFA (10 mL) was slowly added to a solution of 1)-5-fluoro-2-iodobenzoate (5 g, 9 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum. The resulting residue was triturated with petroleum ether for 30 minutes at room temperature. The mixture was filtered and the solid was rinsed with petroleum ether. The precipitate was collected and dried in vacuo to give the title compound (4.1 g, yield: 91%) as a white solid. ESI-MS: mass calculated for C ₁₉ H ₁₇ FIN ₃ O ₄ , 497.0; m/z actual value, 498.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.16 (d, J = 7.3 Hz, 1 H), 7.78 (d, J = 11.0 Hz, 1 H), 7.28 - 7.43 (m, 5 H), 4.60 (s, 2 H), 4.57 (s, 2 H), 3.74 (q, J = 7.2 Hz, 2 H), 1.23 (t, J = 7.2 Hz, 3 H) ppm; ¹⁹ F NMR (376 MHz, DMSO- d ₆ ) δ -119.91 (s, 1 F) ppm.

Step D. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-5-fluo Rho-2-iodobenzoyl chloride.

4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl) in SOCl ₂ (14 mL) A solution of -5-fluoro-2-iodobenzoic acid (3.5 g, 7 mmol) was heated at reflux for 15 minutes. The reaction mixture was cooled to room temperature and concentrated. Anhydrous toluene was added to the residue and the mixture was then evaporated to give the crude product (3.6 g) as a yellow gum, 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 at reflux for 16 hours. The mixture was diluted with petroleum ether and filtered through a short pad of silica gel. The silica gel was washed with hexane. The filtrate was concentrated under reduced pressure at 0-2°C. The residue was purified by silica column chromatography (eluent: 0-10% EtOAc in petroleum ether) to give the title compound (23.3 g, crude) as a colorless liquid. 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.

Add a toluene solution (40 mL) of ethyl 3-methylpent-2-enoate (20 g, crude) dropwise under nitrogen to a toluene solution (1 M) (118 mL, 118 mmol) of DIBAL-H at -78°C. did. The reaction mixture was stirred at -78°C for 2 hours. The mixture was warmed to room temperature and slowly poured into saturated aqueous sodium potassium tartrate solution at 0°C. The mixture was stirred for 2 hours 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, yield of 2 steps: 57%). got it 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 hour. The mixture was filtered through a short pad of ^Celite® . The pad was washed with DCM. The combined filtrates were 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 obtain the title compound (1.5 g, yield: 77%) as a colorless liquid. 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 .

Triethylamine ( After adding 4.6 mL, 33 mmol), a DCM solution of TiCl ₄ (1 M) (5 mL, 5 mmol) was added dropwise. The resulting mixture was stirred at 0° C. for 1 hour, then warmed to room temperature and stirred for 4 hours. The mixture was poured into saturated aqueous NH ₄ Cl solution. The mixture became cloudy and was filtered through a pad of ^Celite® . The pad was washed with EtOAc. The combined filtrates were 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 give the title compound (1.3 g, yield: 70%) as a pale yellow oil.

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 NaBH ₄ (218 mg, 5.8 mmol). ) was added, and 1 hour later, an additional batch of NaBH ₄ (218 mg, 5.8 mmol) was added. Total NaBH ₄ (1.1 g, 29 mmol) was added. The reaction mixture was stirred overnight at room temperature. 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 (430 mg, yield: 33%) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.95 - 7.02 (m, 1 H), 6.84 (ddd, J = 12.2, 8.3, 1.3 Hz, 1 H), 6.52 - 6.63 (m, 1 H), 5.17 - 5.28 (m, 1 H), 3.85 (d, J = 5.6 Hz, 2 H), 3.73 (s, 1 H), 1.91 - 2.07 (m, 2 H), 1.58 - 1.68 (m, 3 H), 0.89 - 0.96 (m, 3 H).

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. did. The mixture was stirred overnight at room temperature. 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 in THF (1 M) (10.6 mL, 13.8 mmol) was added to the mixture under nitrogen. After 45 minutes at -78°C, a solution of hexachloroethane (8.9 g, 37.8 mmol) in THF (20 mL) was added dropwise. The reaction mixture was warmed to room temperature and stirred overnight. The mixture was poured into saturated aqueous NH ₄ Cl solution 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 (1.8 g, yield: 58%) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.52 (dd, J = 10.0, 2.7 Hz, 1 H), 4.07 - 4.15 (m, 1 H), 3.70 (td, J = 11.3, 2.8 Hz, 1 H) , 2.57 (s, 3 H), 2.37 - 2.47 (m, 1 H), 2.11 - 2.19 (m, 1 H), 1.86 - 1.90 (m, 1 H), 1.72 - 1.75 (m, 1 H), 1.64 (d, J = 2.0 Hz, 1 H), 1.53 (d, J = 6.6 Hz, 1 H) ppm.

Step B. 5-Chloro-3-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-amine .

5-Chloro-3-methyl-4-nitro-1-(tetrahydro _- 2H-pyran-2-yl) in MeOH/THF/H 2 O (v/v/v, 1/1/1, 3 mL) To a mixture of -1H-pyrazole (100 mg, 0.4 mmol) 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 hours. The mixture was cooled to room temperature and filtered through a pad of ^Celite® . The pad was washed with EtOAc. The combined filtrates were washed with saturated aqueous NaHCO ₃ solution. 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 (70 mg, yield: 79%) as a yellow oil. ESI-MS: mass calculated for C ₉ H ₁₄ ClN ₃ O, 215.1; m/z actual value, 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.

2-Chloro-3-nitrophenol (200 mg, 1.2 mmol), 2-(( tert -butyldiphenylsilyl)oxy)ethan-1-ol (554 mg, 1.8 mmol) and PPh ₃ in THF (10 mL) To a mixture of (453 mg, 1.7 mmol) was added DEAD (281 mg, 161 mmol) at 0°C under nitrogen. The mixture was warmed to room temperature and stirred at room temperature for 12 hours. Saturated aqueous NH ₄ Cl solution was added and the mixture was extracted with EtOAc. The organics were 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 (240 mg, yield: 46%) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.72 (dd, J = 7.8, 1.5 Hz, 4 H), 7.35 - 7.49 (m, 7 H), 7.30 (t, J = 8.2 Hz, 1 H), 7.12 (dd, J = 8.3, 1.2 Hz, 1 H), 4.20 - 4.25 (m, 2 H), 4.06 (t, J = 4.9 Hz, 2 H), 1.06 (s, 9 H) ppm.

Step B. 3-(2-((tert-butyldiphenylsilyl)oxy)ethoxy)-2-chloroaniline.

tert -butyl(2-(2-chloro-3-nitrophenoxy)ethoxy)diphenylsilane (220 mg, 0.5 mmol) in THF (3 mL), MeOH (3 mL), and H ₂ O (3 mL). , NH ₄ Cl (258 mg, 4.8 mmol) was added to iron powder (269 mg, 4.8 mmol). The reaction mixture was stirred at 70°C for 2 hours. The mixture was cooled to room temperature, diluted with EtOAc and filtered through a pad of ^Celite® . Celite ^® was washed with EtOAc. The combined filtrates were 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 (192 mg, yield: 92%) as a yellow solid. ESI-MS: mass calculated for C ₂₄ H ₂₈ ClNO ₂ Si, 425.2; m/z actual value, 426.1[M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.75 (dd, J = 7.9, 1.6 Hz, 4 H), 7.35 - 7.48 (m, 6 H), 6.97 (t, J = 8.1 Hz, 1 H), 6.42 (dd, J = 8.2, 1.1 Hz, 1 H), 6.33 (dd, J = 8.2, 1.1 Hz, 1 H), 4.13 - 4.17 (m, 2 H), 4.07 - 4.13 (m, 2 H), 4.01 - 4.06 (m, 2 H), 1.06 (s, 9 H) ppm.

Intermediate 9: 5-((benzyloxy)methyl)-4-ethyl-2-(7-methyl-1-oxo-4-(prop-1-en-2-yl)-1H-isochromen-6 -1)-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) was added. The reaction mixture turned orange and was stirred overnight at 50° C. under nitrogen. The mixture was cooled to room temperature, diluted with EtOAc and then 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 (eluent: 0-3% EtOAc in petroleum ether) to give the title compound (900 mg, yield: 72%) as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.57 (d, J = 8.1 Hz, 1 H), 7.24 (d, J = 4.6 Hz, 1 H), 2.22 (d, J = 1.5 Hz, 3 H), 1.58 (s, 9 H) 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 .

tert -Butyl 2-bromo-4-fluoro-5-methylbenzoate (750 mg, 2.6 mmol), 5-((benzyloxy)methyl)-4-ethyl-2,4- in DMF (8 mL) A mixture of dihydro-3H-1,2,4-triazol-3-one (800 mg, 3.4 mmol) and Cs ₂ CO ₃ (1.7 g, 5.2 mmol) was stirred at 90°C for 16 hours. The reaction was quenched by addition of saturated aqueous NH ₄ Cl solution. 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 combi-flash chromatography (SiO ₂ , eluent: 0-22% EtOAc in petroleum ether) to give the title compound (1 g, yield: 71%) as a colorless gum. ESI-MS: mass calculated for C ₂₄ H ₂₈ BrN ₃ O ₄ , 501.1; m/z actual value, 502.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.64 (d, J = 6.1 Hz, 2 H), 7.33 - 7.43 (m, 5 H), 4.61 (s, 2 H), 4.50 (s, 2 H), 3.85 (q, J = 7.3 Hz, 2 H), 2.31 (s, 3 H), 1.62 (s, 9 H), 1.36 (t, J = 7.2 Hz, 3 H) ppm.

Step C. 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-2-bro Parent-5-methylbenzoic acid .

tert -Butyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1- in DCM (5 mL) To a mixture of 1)-2-bromo-5-methylbenzoate (500 mg, 0.90 mmol) was added TFA (1 mL). The mixture was stirred at room temperature for 12 hours. The mixture was concentrated. The residue was dissolved with DCM and petroleum ether was added slowly. The mixture was stirred at room temperature for 30 minutes. The mixture was filtered and the precipitate was rinsed with petroleum ether. The solid was collected and dried in vacuo to give the title compound (360 mg, yield: 86%) as a white solid. ESI-MS: mass calculated for C ₂₀ H ₂₀ BrN ₃ O ₄ , 445.1; m/z actual value, 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 -1)-2,4-dihydro-3H-1,2,4-triazol-3-one .

4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl in DMF (3.9 mL) under nitrogen. )-2-Bromo-5-methylbenzoic acid (560 mg, 1.26 mmol), 3-methylbuta-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) were added Pd(OAc) ₂ (141 mg, 0.63 mmol). The reaction mixture was stirred overnight at 90° C. 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 (410 mg, yield: 73%) as a yellow solid. ESI-MS: mass calculated for C ₂₅ H ₂₅ N ₃ O ₄ , 431.2; m/z actual value, 432.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.28 (s, 1 H), 7.59 (s, 1 H), 7.31 - 7.46 (m, 5 H), 7.17 (s, 1 H), 5.30 - 5.37 (m , 1 H), 5.15 (s, 1 H), 4.63 (s, 2 H), 4.52 (s, 2 H), 3.87 (q, J = 7.1 Hz, 2 H), 2.46 (s, 3 H), 2.10 (s, 3 H), 1.38 (t, J = 7.2 Hz, 3 H) 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 .

(COCl) ₂ (12.7 g, 10.0 mmol) and DMF (69.6 mg, 0.952 mmol) in a solution of 2,6-dichloro-5-fluoronicotinic acid (20 g, 95 mmol) in THF (200 mL) at 0°C. ) was added dropwise. The mixture was stirred at 0°C for 30 minutes, then warmed to 25°C and stirred for 1 hour. 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.

2,6-dichloro-5 in THF (50 mL) at 0° C. 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). -A solution of fluoronicotinoyl chloride (21.7 g, 96.0 mmol) was added. The mixture was stirred at 25°C for 1 hour. The mixture was poured into water (300 mL). The aqueous phase was extracted with ethyl acetate (300 mL). The combined organic phases were 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 obtain the title compound (21 g, 86.82% yield). MS (ESI): mass calculated for C ₉ H ₈ Cl ₂ FNO ₂ , 250.1; m/z actual value, 252.0 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ ) 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 .

3-((benzyloxy)methyl)-4-ethyl-1H-1,2 in a mixture of isopropyl 2,6-dichloro-5-fluoronicotinate (4 g, 15.87 mmol) in DMSO (40 mL) ,4-triazol-5(4H)-one (3.89 g, 16.66 mmol) and K ₂ CO ₃ (3.29 g, 23.80 mmol) were added. The mixture was stirred at 80°C for 3 hours. LCMS showed that the starting material was consumed and the desired mass was detected. The mixture was diluted with H ₂ O (30 mL) and extracted with EtOAc (50 mL x 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 obtain the title compound (5.7 g, 79.86% yield). MS (ESI): mass calculated for C ₂₁ H _{2 2} ClFN ₄ 0 ₄ , 448.1; m/z actual value, 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.

Methyl 2-bromo-4,5-difluorobenzoate (100.0 g, 398 mmol), 5-((benzyloxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2, To a flask charged with 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 and then added 5-((benzyloxy)methyl)-4-ethyl-2,4-dihydro-3H-1,2,4-triazole-3. -one (11 g, 51 mmol) was added. The reaction mixture was continuously stirred 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 minutes, then water (500 mL) was added. The mixture was stirred for 2 hours. 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.

Methyl 4-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1 in dimethylacetamide (300 mL) -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 To a mixture of CO ₃ (42.6 g, 130 mmol) was slowly added 3-methylbutanal (41.4 mL, 386 mmol). The reaction mixture was heated at 80° C. under nitrogen for 22 hours. The mixture was filtered and quenched with aqueous NH ₄ Cl solution until the “pH” changed to 7-8. The mixture was extracted with ethyl acetate (2000 mL x 2). 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 give the title compound (61.7 g, yield: 56%) as an oil. ¹ H NMR (300 MHz, CDCl ₃ ) 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-methylbuta-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 minutes, then warmed to room temperature and stirred overnight. The reaction mixture was diluted with DCM and then washed with saturated aqueous NaHCO ₃ solution and saturated aqueous Na ₂ CO ₃ solution. 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 obtain the title compound (35.8 g, yield: 80%) as a light yellow oil. 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-Methylbuta-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 hours until the mixture turned into a black solution. The mixture was washed with aqueous ammonia solution (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 (650 mg, yield: 26%) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.20 (dt, J = 4.1, 2.0 Hz, 1 H), 2.11 (s, 3 H), 1.81 (d, J = 2.0 Hz, 6 H) ppm.

Example 22 of PCT/IB2020/053601 (published as WO 2020/212897 on October 22, 2020) is 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) Commenced.

Step A. 6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluo Ro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one.

5-((benzyloxy)methyl)-4-ethyl-2-(7-fluoro-3-hydroxy-4-isopropyl-1-oxoisochroman-6-yl)- in AcOH (160 mL) To a mixture of 2,4-dihydro-3H-1,2,4-triazol-3-one (Intermediate 11, 56 g, 123 mmol) was added o-toluidine (14.8 g, 138 mmol). The reaction mixture was heated at 80° C. for 16 hours. The mixture was concentrated and the “pH” was then adjusted to 7-8 using aqueous NaHCO ₃ solution. The mixture was extracted with ethyl acetate (160 mL x 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 (32.5 g, yield: 50%) as an oil. MS (ESI): mass calculated for C ₃₁ H ₃₁ FN ₄ O ₃ , 526.2; m/z actual value, 527.4 [M+H] ⁺ . ¹ H NMR (300 MHz, CDCl ₃ ) 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 .

6-(3-((benzyloxy)methyl)-4-ethyl-5-oxo-4,5-dihydro-1H-1,2,4-triazole-1 in DCM (230 mL) at -78°C To a stirred solution of -1)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinolin-1(2H)-one (26.5 g, 50.3 mmol) was added a DCM solution of BCl ₃ under nitrogen ( 1 M) (290 mL, 290 mL) was added. The reaction mixture was stirred at 15°C for 0.5 hours. The reaction was quenched with MeOH (100 mL) at -78°C to -20°C. The mixture was partitioned between water and DCM. The organic layer was separated and the aqueous layer was extracted with DCM (110 mL x 2). The combined organic extracts were washed with brine (30 mL x 2), dried over anhydrous Na ₂ SO ₄ , 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 pure product (17.5 g), which was recrystallized in ethanol and water to give the title compound (16 g, yield: 73%) as a white solid. ) was obtained. MS (ESI): mass calculated for C ₂₄ H ₂₅ FN ₄ O ₃ , 436.2; m/z actual value, 437.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.33 (d, J = 11.2, 1 H), 8.08 (d, J = 6.8, 1 H), 7.39-7.33 (m, 3 H), 7.28 (s, 1 H), 6.85 (s, 1 H), 4.69 (br s, 2 H), 3.94 (q, J = 7.11 Hz, 2 H), 3.27 (td, J = 13.66, 6.82 Hz, 1 H), 2.32 ( br s, 1 H), 2.17 (s, 3 H), 1.45 (t, J = 7.11 Hz, 3 H), 1.32 (dd, J = 6.82, 1.83 Hz, 6 H) ppm.

It is noted that the compounds of formula (Z) described herein are described in PCT/IB2020/053601 (published as WO 2020/212897 on October 22, 2020), which document is incorporated in its entirety for all purposes. It is incorporated by reference into the specification.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a hypomethylating agent.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a hypomethylating agent.

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

According to certain embodiments, the hypomethylating agent is azacytidine, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, a combination therapy comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a BCL-2 inhibitor is provided.

In some embodiments, a combination therapy consisting of a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a BCL-2 inhibitor is provided.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, and venetoclax, or a pharmaceutically acceptable salt or solvate thereof. .

In some embodiments, combination therapy is provided comprising Compound A , or a pharmaceutically acceptable salt or solvate thereof, and venetoclax, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy comprising Compound A1 and venetoclax, or a pharmaceutically acceptable salt or solvate thereof, is provided.

In some embodiments, combination therapy comprising Compound A2 and venetoclax, or a pharmaceutically acceptable salt or solvate thereof, is provided.

In some embodiments, combination therapy comprising Compound A3 and venetoclax, or a pharmaceutically acceptable salt or solvate thereof, is provided.

In some embodiments, combination therapy is provided comprising Compound A4-a , or a solvate thereof, and venetoclax, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy is provided comprising Compound A4-b , or a hydrate thereof, and venetoclax, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy comprising Compound A4 and venetoclax, or a pharmaceutically acceptable salt or solvate thereof, is provided.

In some embodiments, at least one other antineoplastic agent that is a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapies comprising salts or solvates of

In some embodiments, at least one that is a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and azacytidine. A combination therapy comprising another anti-neoplastic agent, or a pharmaceutically acceptable salt or solvate thereof, is provided.

In some embodiments, Compound A , or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapies comprising pharmaceutically acceptable salts or solvates are provided.

In some embodiments, Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy is offered.

In some embodiments, Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy is offered.

In some embodiments, Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy is offered.

In some embodiments, Compound A4-a , or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapies comprising salts or solvates are provided.

In some embodiments, Compound A4-b , or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt thereof. Alternatively, combination therapy comprising a solvate is provided.

In some embodiments, Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy is offered.

In some embodiments, at least one other antineoplastic agent that is a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapies comprising salts or solvates of

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof, Combination therapies comprising pharmaceutically acceptable salts or solvates are provided.

In some embodiments, Compound A , or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy comprising cargo is provided.

In some embodiments, combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt thereof, and azacitidine, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt thereof, and azacitidine, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt thereof, and azacitidine, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, combination therapy comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof This is provided.

In some embodiments, the combination therapy comprising Compound A4-b , or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and azacytidine, or a pharmaceutically acceptable salt or solvate thereof, provided.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt thereof, and azacitidine, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, at least one other antineoplastic agent that is a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and decitabine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapies comprising salts or solvates are provided.

In some embodiments, at least one agent that is a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and decitabine. Combination therapy comprising other antineoplastic agents, or pharmaceutically acceptable salts or solvates thereof, is provided.

In some embodiments, Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt or solvate thereof. Combination therapy comprising an acceptable salt or solvate is provided.

In some embodiments, a combination comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt or solvate thereof. Therapy is provided.

In some embodiments, a combination comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt or solvate thereof. Therapy is provided.

In some embodiments, a combination comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt or solvate thereof. Therapy is provided.

In some embodiments, Compound A4-a , or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt thereof. Alternatively, combination therapy comprising a solvate is provided.

In some embodiments, Compound A4-b , or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt thereof, or Combination therapies comprising solvates are provided.

In some embodiments, a combination comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is decitabine, or a pharmaceutically acceptable salt or solvate thereof. Therapy is provided.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a DNA intercalating agent.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein 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 comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is an anthracycline.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is an anthracycline.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is an anthracycline. This is provided.

In some embodiments, combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is an anthracycline.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a pyrimidine analog.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein 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 comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is cytarabine.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is cytarabine.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is cytarabine. This is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is cytarabine.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a purine analog.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein 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 comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is fludarabine.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is fludarabine.

In some embodiments, a combination comprising Compound A , or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is fludarabine.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is an IDH inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein 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 comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is ivosidenib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is ivosidenib.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is ivosidenib. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ivosidenib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is an isocitrate dehydrogenase-2 inhibitor (e.g., enasidenib).

According to an embodiment, the IDH inhibitor is enasidenib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is enasidenib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is enasidenib.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is enasidenib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is an immunomodulatory antineoplastic agent.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is an immunomodulatory antineoplastic agent, a PD-1 inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is an immunomodulatory antineoplastic agent.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is an immunomodulatory antineoplastic agent that is a PD-1 inhibitor.

According to an embodiment, the immunomodulatory antineoplastic agents are nivolumab, atezolizumab and pembrolizumab, thalidomide, lenalidomide, pomalidomide, Bacillus Calmette-Guerin (BCG) and levamisole.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is nivolumab.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is nivolumab.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab. This is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is nivolumab.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is atezolizumab.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is atezolizumab.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is atezolizumab. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is atezolizumab.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is pembrolizumab.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is pembrolizumab.

In some embodiments, a combination comprising Compound A , or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pembrolizumab.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is thalidomide.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is thalidomide.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is thalidomide provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is thalidomide.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is lenalidomide.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is lenalidomide.

In some embodiments, Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is lenalidomide. Combination therapy is offered.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide. .

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lenalidomide.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is pomalidomide.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is pomalidomide.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is pomalidomide. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is pomalidomide.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is BCG.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is BCG.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is BCG provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is BCG.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is levamisole.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is levamisole.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole. This is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is levamisole.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a DHODH inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a DHODH inhibitor.

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

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

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising, wherein at least one other antineoplastic agent is a DHODH inhibitor of formula (Z), a pharmaceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof.

In some embodiments, Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt thereof Combination therapy comprising at least one other antineoplastic agent that is an isotope, N-oxide, solvate, or stereoisomer is provided.

In some embodiments, Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N-oxide, solvate thereof, or combination therapy comprising at least one other antineoplastic agent that is a stereoisomer.

In some embodiments, Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N-oxide, solvate thereof, or combination therapy comprising at least one other antineoplastic agent that is a stereoisomer.

In some embodiments, Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N-oxide, solvate thereof, or combination therapy comprising at least one other antineoplastic agent that is a stereoisomer.

In some embodiments, Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N Combination therapy comprising at least one other antineoplastic agent that is an oxide, solvate, or stereoisomer is provided.

In some embodiments, Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N- Combination therapy comprising at least one other antineoplastic agent that is an oxide, solvate, or stereoisomer is provided.

In some embodiments, Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof and a DHODH inhibitor of Formula (Z), a pharmaceutically acceptable salt, isotope, N-oxide, solvate thereof, or combination therapy comprising at least one other antineoplastic agent that is a stereoisomer.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a kinase inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a kinase inhibitor.

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

In some embodiments, the kinase inhibitor is an inhibitor of FLT3 and/or BTK.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a FLT3 inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a FLT3 inhibitor.

According to embodiments, the FLT3 inhibitor is selected from the group consisting of sorafenib, sunitinib, midostaurin (PKC412), lestaurtinib (CEP-701), tandutinib (MLN518), quizartinib (AC220), gilteritinib (ASP2215), or It is KW-2449.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is sorafenib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is sorafenib.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib. This is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sorafenib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is sunitinib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is sunitinib.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is sunitinib. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is sunitinib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is midostaurin.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is midostaurin.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin. This is provided.

In some embodiments, combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is midostaurin.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is lestaurtinib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is lestaurtinib.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is lestaurtinib. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is lestaurtinib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is tandutinib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is tandutinib.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib. This is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is tandutinib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is AC220.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is AC220.

In some embodiments, the combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is AC220 provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is AC220.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is ASP2215.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is ASP2215.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is ASP2215 provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ASP2215.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is KW-2449.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is KW-2449.

In some embodiments, a combination comprising Compound A , or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is KW-2449.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a BTK inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a BTK inhibitor.

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor. This is provided.

In some embodiments, combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a BTK inhibitor.

According to an embodiment, the BTK inhibitor is ibrutinib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is ibrutinib.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is ibrutinib.

In some embodiments, a combination comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is ibrutinib. Therapy is provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is ibrutinib.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is a CD20 inhibitor.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is a CD20 inhibitor.

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

In some embodiments, combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is a CD20 inhibitor. This is provided.

In some embodiments, combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is a CD20 inhibitor.

In some embodiments, a combination therapy is provided comprising a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, a BCL-2 inhibitor, and at least one other antineoplastic agent, wherein at least One other antineoplastic agent is GA101.

In some embodiments, a menin-MLL inhibitor of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent. Combination therapy is provided comprising: wherein at least one other antineoplastic agent is GA101.

In some embodiments, a combination therapy comprising Compound A or a pharmaceutically acceptable salt or solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof and at least one other antineoplastic agent that is GA101 provided.

In some embodiments, a combination therapy is provided comprising Compound A1 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

In some embodiments, a combination therapy is provided comprising Compound A2 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

In some embodiments, a combination therapy is provided comprising Compound A3 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

In some embodiments, a combination therapy is provided comprising Compound A4-a or a solvate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

In some embodiments, a combination therapy is provided comprising Compound A4-b or a hydrate thereof, venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

In some embodiments, a combination therapy is provided comprising Compound A4 , venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one other antineoplastic agent that is GA101.

All possible combinations of the embodiments shown above are considered to be included within the scope of the present invention.

In some embodiments, methods of treating a subject diagnosed with a hematopoietic disorder are provided. The present invention provides, e.g., to a subject, a therapeutically effective amount of a menin-MLL inhibitor as described herein; and a therapeutically effective amount of a BCL-2 inhibitor; and optionally administering a therapeutically effective amount of at least one other antineoplastic agent.

A further embodiment of the invention relates to a method as described herein wherein a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof is orally administered to a subject.

A further embodiment of the present invention is a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, administered to the subject at a dose of about 1 mg/kg to about 50 mg/kg. It relates to a method as described.

A further embodiment of the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, wherein the compound of formula (I) is administered to the subject at a dose of about 2.5 mg/kg to about 25 mg/kg. It relates to a method as described.

A further embodiment of the invention provides a method for administering a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, to the subject at a dose of about 7.5 mg/kg to about 12.5 mg/kg. It relates to a method as described.

A further embodiment of the present invention provides a method for preparing a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, wherein the compound is administered to the subject at a dose of about 8 mg/kg to about 10 mg/kg. It relates to a method as described.

A further embodiment of the invention provides a method for administering a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, to a subject, wherein the compound is administered to the subject at a dose of about 0.1 mg/kg to about 5 mg/kg. It relates to a method as described.

A further embodiment of the invention relates to a method as described herein wherein the BCL-2 inhibitor is orally administered to the subject.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject at a dose of about 1 mg/kg to about 50 mg/kg.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject at a dose of about 2.5 mg/kg to about 25 mg/kg.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject at a dose of about 7.5 mg/kg to about 12.5 mg/kg.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject at a dose of about 8 mg/kg to about 10 mg/kg.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject at a dose of about 0.1 mg/kg to about 5 mg/kg.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered intravenously or subcutaneously to the subject.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 10 mg/m ² to about 250 mg/m ² will be.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 50 mg/m ² to about 150 mg/m ² will be.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 60 mg/m ² to about 100 mg/m ² will be.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered intravenously or subcutaneously to the subject at a dose of about 75 mg/m ² .

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is orally administered to the subject in a dose of about 1 mg to about 500 mg.

Additional embodiments of the invention include cancer (e.g., the cancer is myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small lymphocytic lymphoma (SLL), or chronic lymphocytic lymphoma. A method of treating a subject diagnosed with leukemia (CLL), said method comprising:

A therapeutically effective amount of a menin-MLL inhibitor of formula (I), or a tautomeric or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof; and

comprising administering a therapeutically effective amount of a DHODH inhibitor of Formula (Z), or a pharmaceutically acceptable salt, isotope, N-oxide, solvate, or stereoisomer thereof;

For example, wherein the menin-MLL inhibitor and the DHODH inhibitor may be administered according to a dosing schedule for a period of time, e.g., (i) 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); administered simultaneously or sequentially on the same day(s) within (ii) a period, or a one-year period, etc.), and/or on different days within (ii) a period.

Additional embodiments of the invention include cancer (e.g., the cancer is myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small lymphocytic lymphoma (SLL), or chronic lymphocytic lymphoma. A method of treating a subject diagnosed with leukemia (CLL), said method comprising:

A therapeutically effective amount of Compound A or a pharmaceutically 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 solvate thereof) hydrate, or a menin-MLL inhibitor, which is Compound A4 ); and

A therapeutically effective amount of Compound 22 or a pharmaceutically acceptable salt, solvate, stereoisomer, isotopic variant, or N-oxide thereof (e.g., 6-(4-ethyl-3-(hydroxymethyl)-5 -Oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl)-7-fluoro-4-isopropyl-2-(o-tolyl)isoquinoline-1(2H) -on) comprising administering a DHODH inhibitor;

For example, wherein the menin-MLL inhibitor and the DHODH inhibitor may be administered according to a dosing schedule for a period of time, e.g., (i) 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); administered simultaneously or sequentially on the same day(s) within (ii) a period, or a one-year period, etc.), and/or on different days within (ii) a period.

A further embodiment of the invention relates to a method as described herein, wherein a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, is administered daily to the subject.

A further embodiment of the invention relates to a method as described herein, wherein a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, is administered to the subject for at least 7 days.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered daily to the subject.

A further embodiment of the invention relates to a method as described herein, wherein the BCL-2 inhibitor is administered to the subject for at least 7 days.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered daily to the subject.

A further embodiment of the invention relates to a method as described herein, wherein at least one other antineoplastic agent is administered to the subject for at least 21 days.

The optimal dosage of any of the therapeutic compounds described herein can be readily determined and will vary depending on the particular compound used, the mode of administration, the strength of the formulation, and the progression of the disease, syndrome, condition or disorder. Additionally, depending on factors associated with the particular subject being treated, including subject sex, age, weight, diet, and time of administration, the dosage will need to be adjusted to achieve an appropriate therapeutic level and desired therapeutic effect.

Therefore, the above dosages are examples of average cases. Of course, there may be individual cases where a larger or smaller dosage range would be beneficial, and such cases are within the scope of the present invention.

Whenever use of a therapeutic compound described herein is administered to a subject in need thereof, the therapeutic compound of the invention may be administered in any of the compositions and administration regimens described above, or in compositions and administration regimens established in the art. It can be administered by.

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 administration is simultaneous, the combination may be administered in the same or 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, a BCL-2 inhibitor can be administered before, concurrently with, or after administration of other optional antineoplastic agents. Adjunctive therapy, i.e., where one or two agent(s) are used as primary treatment and another agent is used to support the primary treatment, is also an embodiment of the invention.

One embodiment of the invention provides a therapeutically effective amount of a BCL-2 inhibitor; and optionally, compounds of formula (I) and pharmaceutically acceptable salts, and solvates thereof, for use in combination with a therapeutically effective amount of at least one other antineoplastic agent, or as mentioned in any of the embodiments. It relates to a therapeutically effective amount of a menin-MLL inhibitor, including any subgroup thereof. In certain embodiments, the therapeutically effective amount described above is administered in a separate dosage form for use in treating a subject diagnosed with a hematopoietic disorder.

One embodiment of the invention is a combined preparation for simultaneous, separate or sequential use in treating a subject diagnosed with a hematopoietic disorder, comprising: a menin-MLL inhibitor of formula (I) and a pharmaceutically acceptable salt thereof; and solvates, or any subgroup thereof as mentioned in any of the embodiments, and BCL-2 inhibitors; and, optionally, at least one other antineoplastic agent.

The following examples are provided to illustrate some of the concepts described in this disclosure. The examples are considered to provide embodiments, but should not be considered limiting to the more general embodiments described herein.

Example

General synthesis scheme

Venetoclax

Venetoclax is available for purchase.

Azacitidine

Azacitidine is available commercially.

Compounds of Formula I

In this section, as in all other sections unless the context indicates otherwise, references to formula (I) also include all other subgroups and examples thereof as defined herein.

The general preparation of some typical examples of compounds of compound (I) is described below and in specific examples, generally from starting materials that are commercially available or prepared by standard synthetic processes commonly used by those skilled in the art of organic chemistry. It is manufactured with The following reaction scheme is merely intended to illustrate an example of the present invention and is not intended to limit the present invention in any way.

Alternatively, compounds of formula (I) can also be prepared by reaction protocols similar to those described in the general schemes below, combined with standard synthetic procedures commonly used by those skilled in the art.

In the reactions described in the schemes, although this is not always explicitly indicated, when reactive functional groups (e.g. hydroxy, amino, or carboxy groups) are desired in the final product, they are added to avoid their unwanted participation in the reaction. Those skilled in the art will recognize that protection may be necessary. In general, conventional protecting groups (PG) may be used according to standard practice. Protecting groups may be removed at convenient subsequent steps using methods known in the art.

Those skilled in the art will recognize that, in the reactions described in the schemes, it may be advisable or necessary to carry out the reactions under an inert atmosphere , for example an N ₂ -gas atmosphere .

It may be necessary to cool the reaction mixture prior to reaction work-up (refers to the series of operations required to isolate and purify the product(s) of a chemical reaction, such as quenching, column chromatography, extraction, etc.). This will be clear to those skilled in the art.

Those skilled in the art will recognize that heating the reaction mixture with agitation can improve reaction results. In some reactions, microwave heating may be used instead of conventional heating to shorten the overall reaction time.

Those skilled in the art will recognize that other sequences of chemical reactions shown in the schemes below can also lead to the desired compounds of formula (I).

Those skilled in the art will appreciate that the intermediate and final compounds shown in the schemes below can be further functionalized according to methods well known to those skilled in the art. The intermediates and compounds described herein can be isolated in free form or as salts or solvates thereof. The intermediates and compounds described herein can be synthesized in the form of mixtures of tautomeric and stereoisomeric forms, which can be separated from each other according to resolution procedures known in the art.

All abbreviations used in the general scheme for Formula (I) are as defined in Table 1b below in the Examples part. Variables are as defined within the scope or as specifically defined in the general reaction equation.

Part A) Schemes 1a, 1b, 1c, 2a, 2b and 3

Scheme 1a

Scheme 1b

Scheme 1c

In Schemes 1a, 1b and 1c, the following reaction conditions apply:

Step 1: At a suitable temperature, for example -70°C, in the presence of a suitable base, for example, TMEDA, and a suitable organometallic reagent, for example, isopropylmagnesium bromide, in a suitable solvent, for example, THF. ;

Step 2: at a suitable temperature, for example from 0° C. to room temperature, in the presence of a suitable oxidizing reagent, for example DMP, in a suitable solvent, for example DCM;

Step 3: at a suitable temperature, for example from -20° C. to room temperature, in the presence of a suitable organometallic reagent, for example isopropylmagnesium bromide, in a suitable solvent, for example THF;

Step 4: At a suitable temperature, for example 80° C., in the presence of a suitable base, for example NaOH, in a suitable solvent, for example THF and H ₂ O;

Step 5: At a suitable temperature, for example room temperature, in the presence of a suitable amide condensation reagent, for example EDCI and HOBt, in the presence of a suitable base, for example NMM, in a suitable solvent, for example DCM. ;

Step 6: at a suitable temperature, for example -70° C., in the presence of a suitable organometallic reagent, for example isopropyllithium, in a suitable solvent, for example THF;

Step 7: At a suitable temperature, for example 90° C., in the presence of a suitable organometallic catalyst, for example Pd(dppf)Cl ₂ , in the presence of a suitable base, for example Na ₂ CO ₃ , for example In suitable solvents such as 1,4-dioxane and H ₂ O;

Step 8: At a suitable temperature, for example from 0° C. to room temperature, in the presence of a suitable Lewis acid, for example BBr ₃ , in a suitable solvent, for example DCM;

Step 9: At a suitable temperature, for example from -78° C. to 40° C., especially from 0° C. to room temperature, in the presence of a suitable base, for example TEA, DBU or K ₂ CO ₃ , for example DCM, THF or In a suitable solvent such as DMF;

Scheme 2a

Scheme 2b

In Schemes 2a and 2b, the following reaction conditions apply:

Step 9: Refer to Step 9 of Scheme 1;

Step 10: At a suitable temperature, for example room temperature, in the presence of a suitable catalyst, for example Pd/C, in the presence of a suitable reducing reagent, for example H ₂ , optionally in the presence of a suitable reducing reagent, for example TEA. In the presence of a base, for example in a suitable solvent such as THF; Alternatively, at a suitable temperature, such as room temperature, in the presence of a suitable catalyst, for example Pd(dppf)Cl ₂ ·DCM complex, a suitable reducing agent, such as NaBH ₄ , a suitable base, for example TMEDA, for example In a suitable solvent such as THF.

Step 11: For N deprotection, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example TFA, in a suitable solvent, for example DCM; For O deprotection, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example 4-methylbenzenesulfonic acid, in a suitable solvent, for example MeOH;

Step 12: At a suitable temperature, for example 80° C., optionally in the presence of a suitable Lewis acid, for example ZnCl ₂ , in the presence of a suitable reducing reagent, for example NaBH ₃ CN, for example with MeOH. In a suitable solvent such as;

Step 13: At a suitable temperature, for example room temperature, in the presence of a suitable organometallic catalyst, for example Ag(Phen) ₂ OTf, for example 1,3-dibromo-1,3,5-triane In the presence of a suitable brominating agent such as jinan-2,4,6-trione, for example in a suitable solvent such as DCE;

Step 14: At a suitable temperature, for example room temperature, in the presence of a suitable chlorinating reagent, for example oxalyl chloride, in the presence of DMF, in a suitable solvent, for example DCM.

Scheme 3

In Scheme 3, the following reaction conditions apply:

Steps 11-12: See steps 11-12 of Scheme 2;

Step 15: at a suitable temperature, for example 80° C., in the presence of a suitable base, for example Cs ₂ CO ₃ , in a suitable solvent, for example DMF;

Step 16: At a suitable temperature, for example 40° C., in the presence of a suitable base, for example ammonia, in a suitable solvent, for example 1,4-dioxane.

Part B) Schemes 4, 5, 6, 7, 8, 9, 10, 11 and 12

Scheme 4

In Scheme 4, the following reaction conditions apply:

Step 1: At a suitable temperature, for example 90° C., in the presence of a suitable organometallic catalyst, for example Pd(dppf)Cl ₂ , in the presence of a suitable base, for example Na ₂ CO ₃ , for example In suitable solvents such as 1,4-dioxane and H ₂ O;

Step 2: at a suitable temperature, for example room temperature, in the presence of a suitable amide condensation reagent, for example HATU, in the presence of a suitable base, for example DIEA, in a suitable solvent, for example DCM;

Step 3: in a suitable solvent, for example DCM, in the presence of a suitable Lewis acid, for example BBr ₃ , at a suitable temperature, for example -78° C. to room temperature;

Step 4: At a suitable temperature, for example -78°C to 40°C, especially 0°C to room temperature, in the presence of a suitable base, for example TEA, DBU or K ₂ CO ₃ , for example DCM, THF or In a suitable solvent such as DMF;

Step 5: At a suitable temperature, for example room temperature, in the presence of a suitable base, for example LiOH·H ₂ O, in a suitable solvent, for example THF and H ₂ O;

Step 6: At a suitable temperature, for example room temperature, in the presence of a suitable organometallic catalyst, for example Ag(Phen) ₂ OTf, for example 1,3-dibromo-1,3,5-triazine In the presence of a suitable brominating agent such as jinan-2,4,6-trione, for example in a suitable solvent such as DCE;

Step 7: 2 as solvent in the presence of a suitable brominating agent, such as 1,3-dibromo-1,3,5-triazin-2,4,6-trione, at a suitable temperature, for example room temperature. , in the presence of 2,2-trifluoroethan-1-ol.

Scheme 5

In Scheme 5, the following reaction conditions apply:

Step 8: At a suitable temperature, for example -78°C to 40°C, especially 0°C to room temperature, in the presence of a suitable base, for example TEA, DBU or K ₂ CO ₃ , for example DCM, THF or In a suitable solvent such as DMF;

Step 9: At a suitable temperature, for example from -78° C. to 40° C., especially from 0° C. to room temperature, in the presence of a suitable base, for example TEA, DBU or K ₂ CO ₃ , for example DCM, THF or In a suitable solvent such as DMF;

Step 10: H ₂ atmosphere in a suitable solvent, for example MeOH, in the presence of a suitable organometallic catalyst, for example Pd/C, and a suitable base, for example TEA, at a suitable temperature, for example room temperature. under;

Step 11: If PG is Boc, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example TFA, in a suitable solvent, for example DCM.

Scheme 6

In Scheme 6, the following reaction conditions apply:

Step 12: Reductive amination conditions, for example at a suitable temperature such as room temperature to 80° C., in the presence of a suitable Lewis acid such as ZnCl ₂ or an acid such as AcOH, for example NaBH ₃ CN. In the presence of a reducing agent, for example in a suitable solvent such as MeOH;

Step 13: At a suitable temperature, for example 0° C., in the presence of a suitable nucleophile, for example MsCl, in the presence of a suitable base, for example TEA, in a suitable solvent, for example DCM;

Step 14: At a suitable temperature, for example from 0° C. to room temperature, in the presence of a suitable oxidizing agent, for example DMP, in a suitable solvent, for example DCM;

Step 15: At a suitable temperature, for example 50° C., in the presence of a suitable acid, for example HCl, in a suitable solvent, for example ACN;

Step 16: At a suitable temperature, for example room temperature, in the presence or absence of a suitable base, for example TEA, in a suitable solvent, for example THF.

Scheme 7

In Scheme 7, the following reaction conditions apply:

Step 11: If PG is Boc, at a suitable temperature, for example room temperature, in the presence of a suitable acid, for example TFA, in a suitable solvent, for example DCM;

Step 12: At a suitable temperature, for example from room temperature to 80° C., in the presence of a suitable Lewis acid, for example ZnCl ₂ or an acid such as AcOH, in the presence of a suitable reducing agent, for example NaBH ₃ CN, for example In a suitable solvent such as MeOH;

Step 17: At a suitable temperature, for example from room temperature to 80° C., in the presence of a suitable base, for example DIEA or Cs ₂ CO ₃ , in a suitable solvent, for example DCM or DMF;

Step 18: At a suitable temperature, for example 40° C., in the presence of a suitable base, for example ammonia, in a suitable solvent, for example 1,4-dioxane.

Scheme 8

In Scheme 8, the following reaction conditions apply:

Step 9: At a suitable temperature, for example from -78° C. to 40° C., especially from 0° C. to room temperature, in the presence of a suitable base, for example TEA, DBU or K ₂ CO ₃ , for example DCM, THF or In a suitable solvent such as DMF;

Step 10: At a suitable temperature, for example room temperature, in the presence of a suitable organometallic catalyst, for example Pd/C, optionally in the presence of a suitable base, for example TEA, in the presence of a suitable base, for example MeOH. under H ₂ atmosphere in solvent;

Step 19: At a suitable temperature, for example room temperature, in the presence of a suitable chlorinating reagent, for example oxalyl chloride, in the presence of DMF, in a suitable solvent, for example DCM;

Step 20: At a suitable temperature, for example 90° C., in the presence of a suitable nucleophile amine, in a suitable solvent, for example EtOH;

Step 21: At a suitable temperature, for example room temperature, in the presence of a suitable acid, for example HCl in dioxane, in a suitable solvent, for example MeOH;

Step 22: A suitable organometallic catalyst, for example tetrakis(triphenylphosphine)palladium(0), in the presence of a suitable boron reagent, for example trimethylboroxine, at a suitable temperature, for example 110° C. in the presence of a suitable base, for example K ₂ CO ₃ , in a suitable solvent, for example 1,4-dioxane;

Scheme 9

In Scheme 9, the following reaction conditions apply:

Step 23: at a suitable temperature, for example -78°C to -25°C, in the presence of a suitable base, for example DIEA and n-BuLi, in a suitable solvent, for example THF;

Step 24: At a suitable temperature, for example between -65°C and -55°C, in the presence of a suitable reducing agent, for example DIBAL-H, in a suitable solvent, for example toluene, preferably in a suitable flow chemistry system. Carried out in;

Step 25: First, at a suitable temperature, for example from -10° C. to 10° C., in the presence of a suitable base, for example DMAP, in the presence of a suitable condensing agent, for example DCC, for example DCM. In a suitable solvent; Then, at a suitable temperature, for example from -10° C. to 0° C., in the presence of a suitable acid, for example AcOH, in the presence of a suitable reducing agent, for example NaBH ₄ , in a suitable solvent, for example DCM. at;

Step 26: heated to reflux, for example in a suitable solvent such as toluene;

Step 27: at a suitable temperature, for example from -5° C. to 5° C., in the presence of a suitable reducing agent, for example LiBH ₄ , in a suitable solvent, for example 2-methyltetrahydrofuran;

Step 28: at a suitable temperature, for example between 15° C. and 25° C., in the presence of a suitable reducing agent, for example NaBH(OAc) ₃ , in a suitable solvent, for example DCM;

Step 29: At a suitable temperature, for example 15° C. to 25° C., in the presence of a suitable acid, such as HCl, in a suitable solvent, for example IPA;

Step 30: At a suitable temperature, for example 5° C. to 30° C., in the presence of a suitable base, for example TEA, in the presence of a suitable reducing agent, for example NaBH(OAc) ₃ , for example with toluene. In a suitable solvent such as;

Step 31: at a suitable temperature, for example between 50° C. and 55° C., in the presence of a suitable base, for example K ₂ HPO ₄ , in a suitable solvent, for example H ₂ O;

Step 32: If PG is Bn, a suitable catalyst, for example palladium hydroxide on carbon, under hydrogen atmosphere at a suitable temperature, for example -5°C to 45°C, in a suitable pressure range, for example, 0.27 to 0.40 MPa. in a suitable solvent, such as EtOH, in the presence of a suitable acid, for example MSA;

Step 33: In a suitable solvent such as 2-methyltetrahydrofuran, for example in the presence of a suitable base such as TEA, at a suitable temperature such as -50°C to -40°C;

Step 34: In a suitable solvent, such as 2-methyltetrahydrofuran, in the presence of a suitable base, for example TMG, at a suitable temperature, for example 20° C. to 30° C.;

Step 35: MeOH and In a suitable solvent such as;

Alternatively, a suitable catalyst such as 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex, a suitable reducing agent such as sodium borohydride, at a suitable temperature, such as room temperature. , for example in the presence of a suitable base, such as N,N,N',N' -tetramethylenediamine, in a suitable solvent, for example tetrahydrofuran.

Scheme 10

In general, compounds of formula (I), i.e. compounds of formula (Ia), in which Y ¹ is defined as -CH ₂ - and R ² is defined as W ¹ , can be prepared according to Scheme 10 below. In Scheme 10, W ¹ represents chloro, bromo or iodo; All other variables are defined according to the scope of the present invention.

Scheme 10

In Scheme 10, the following reaction conditions apply:

Step 36: At a suitable temperature ranging from 60° C. to 100° C., palladium acetate (Pd(OAc) ₂ ) or tris(dibenzylideneacetone)dipalladium(0)(Pd ₂ (dba) ₃ ) or tetrakis( In the presence of a suitable catalyst, such as triphenylphosphine)palladium(0), in a suitable solvent, for example tetrahydrofuran or dioxane.

Those skilled in the art will recognize that similar chemistry as reported in Step 10 of Scheme 5 and Steps 20, 21 and 22 of Scheme 8 can be performed, starting with compound (Ia).

Scheme 11

In general, compounds of formula (I), i.e. compounds of formula (Ib), wherein Y ¹ is defined as -CR ^5a R ^5b - and R ² is defined as W ¹ , can be prepared according to Scheme 11 below. In Scheme 11, 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.

Scheme 11

In Scheme 11, the following reaction conditions apply:

Step 37: In the presence of a suitable catalyst, such as palladium acetate (Pd(OAc) ₂ ), at a suitable temperature in the range of 80° C. to 200° C., in the presence of a suitable ligand, for example triphenylphosphine or tricyclohexylphosphine. in a suitable solvent, for example dioxane, preferably under sealed conditions, optionally under microwave irradiation.

Those skilled in the art will recognize that similar chemistry as reported in Step 10 of Scheme 5 and Steps 20, 21 and 22 of Scheme 8 can be performed, starting with compound (Ib).

Scheme 12

In Scheme 12, the following reaction conditions apply:

Step 38: at a suitable temperature, for example from room temperature to 80° C., in the presence of a suitable base, for example DIEA, Cs ₂ CO ₃ or DBU, in a suitable solvent, for example DCM, THF or DMF;

Alternatively, at a suitable temperature, for example from room temperature to 100° C., in the presence of a suitable catalyst, for example Pd ₂ dba ₃ , in the presence of a suitable ligand, for example Xantphos, Cs ₂ CO ₃ or Na ₂ in a suitable solvent such as dioxane or a mixture of dioxane and water in the presence of a suitable base such as CO ₃ .

Those skilled in the art will recognize that similar chemistry as reported can be performed starting from intermediate Z where Y ¹ represents O.

When appropriate functional groups are present, compounds of various formulas, or any intermediates used in their preparation, can be further derivatized by one or more standard synthetic methods using condensation, substitution, oxidation, reduction, or cleavage reactions. It will be acknowledged. Particular substitution approaches include conventional alkylation, arylation, heteroarylation, acylation, sulfonylation, halogenation, nitration, formylation, and coupling procedures.

Compounds of formula (I) can be synthesized in the form of racemic mixtures of enantiomers, which can be separated from each other according to resolution procedures known in the art. Racemic compounds of formula (I) containing a basic nitrogen atom can be converted to the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Subsequently, the diastereomeric salt forms are separated, for example by selective or fractional crystallization, and the enantiomers are liberated therefrom by alkali. An alternative way to separate enantiomeric forms of compounds of formula (I) involves liquid chromatography using chiral stationary phases. The pure stereochemically isomeric forms can also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, so long as the reaction occurs stereospecifically.

In the preparation of compounds of formula (I), protection of remote functional groups (e.g. primary or secondary amines) of the intermediate may be necessary. The need for such protection will vary depending on the nature of the remote functional group and the conditions of the manufacturing method. Suitable amino-protecting groups (NH-Pg) include acetyl, trifluoroacetyl, t-butoxycarbonyl (Boc), benzyloxycarbonyl (CBz), and 9-fluorenyl-methyleneoxycarbonyl (Fmoc). Includes. The need for such protection is readily determined by those skilled in the art. For a general description of protecting groups and their uses, 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 examples below. Unless otherwise stated, all starting materials were obtained from commercial suppliers and were used without further purification or, alternatively, can be synthesized by those skilled in the art using well-known methods.

[Table 1b]

'에 적용될 것이다.As will be understood by those skilled in the art, compounds synthesized using the protocols as indicated may exist as solvates, such as hydrides, and/or may contain residual solvents or minor amounts of impurities. A compound or intermediate isolated as a salt form may be integer stoichiometric, i.e., mono- or di-salt, or intermediate stoichiometric. If an intermediate or compound in the following experimental part is indicated as 'HCl salt' without indicating the equivalent number of HCl, this means that the equivalent number of HCl has not been determined. The same principle also applies to all other salt forms referred to in the experimental part, e.g. 'oxalate salt', 'formate salt', or '

' will be applied to.

Even when not explicitly stated in the following experimental protocols, those skilled in the art will recognize that typically after column chromatography purification, the desired fractions are collected and the solvent is evaporated.

If stereochemistry is not indicated, this means that it is a mixture of stereoisomers, unless otherwise indicated or clear from context.

When a stereocenter is indicated with 'RS', this means that a racemic mixture was obtained at the indicated center, unless otherwise indicated.

Example 1 - ( R ) -N -ethyl-5-fluoro- N -isopropyl-2-((5-(2-(6-((2-methoxyethyl) (methyl)amino)-2- Synthesis of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method A

Intermediate 1 - Preparation of tert-butyl (5-methyl-4-oxohexyl)carbamate

Isopropylmagnesium bromide in 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 at -70°C. Solution (19 mL, 55 mmol, 2.9 M in 2-methyltetrahydrofuran) was added slowly and the resulting mixture was slowly warmed to room temperature and stirred for 12 hours. The mixture was poured into saturated aqueous NH ₄ Cl (50 mL) solution and extracted with EtOAc (50 mL × 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) was obtained as a yellow oil.

Intermediate 13 - Preparation of tert-butyl 6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro [3.4]octane-2-carboxylate

tert-butyl 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 at 0°C. 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 hour. 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 FCC on silica gel (PE/EtOAc = 1:0 to 3:1). Purification gave the title intermediate (12.0 g, 58% yield) as a yellow solid.

Intermediate 27 - Preparation of N-ethyl-5-fluoro-N-isopropyl-2-methoxybenzamide

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 at 0°C, HATU (21.5 g, 56.5 mmol) and DIEA (9.10 g, 70.4 mmol) were added slowly in portions. The resulting mixture was slowly warmed to room temperature and stirred for 8 hours. 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% to 20%) to give the title intermediate (12.0 g, 96% yield) as a white solid.

Intermediate 28 - Preparation of N-ethyl-5-fluoro-2-hydroxy-N-isopropylbenzamide

_BBr 3 ( 14.4 mL, 152 mmol) was slowly added, and the resulting mixture was slowly warmed to room temperature and stirred for 8 hours. 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 %) to give the title intermediate (9.0 g, 78% yield) as a white solid.

Alternative Preparation of Intermediate 28

A mixture of 5-fluoro-2-hydroxy-benzoic acid (14.0 kg, 89.68 mol, 1.0 equiv) in THF (168 L, 12 volumes) was adjusted to 15-25° C. and 1,1-carbonyldiimidazole was added. (17.45 kg, 107.62 mol, 1.2 equiv) was added over a period of 1 hour. After addition, the mixture was stirred at 15-25° C. for 18 hours. At this time, N-ethylpropan-2-amine (14.85 kg, 170.39 mol, 1.9 equivalent) was added to the mixture over a period of 2 hours at 15 to 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 aqueous 10% H ₂ SO ₄ (140 kg, 10 vol) and the layers were separated. The organic phase was concentrated to 42-56 L maintaining the temperature below 40°C, and then n-heptane (43 kg, 4.5 volumes) was added to the mixture over a period of 3 hours at 15-25°C. The mixture was then cooled to 0-10° C. and stirred for a further 6 hours. The resulting slurry was filtered and the cake was washed with a tert-butyl methyl ether (MTBE):n-heptane mixture (2:3 volume/volume mixture of 25 kg of MTBE:n-heptane, 2.5 volumes). The cake was washed and repeated two additional times, and the resulting solid was dried in vacuum at 50° C. to obtain intermediate 28 (16.5 kg, purity: 99.1%, yield: 80.4%).

Intermediate 14 - tert-Butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5-yl) -Manufacture of 2,6-diazaspiro[3.4]octane-2-carboxylate

tert -Butyl 6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate in THF (120 mL) (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) mixture was stirred at 25°C for 8 hours. 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) The title intermediate (14.0 g, 73% yield) was obtained as a green solid.

Intermediate 2 - tert-Butyl 6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluoro-phenoxy)-1,2,4-triazine-5-yl)-2, Preparation of 6-diazaspiro[3.4]octane-2-carboxylate

Synthetic Method A for Intermediate 2:

tert -Butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5 in THF (500 mL) -yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 14 ) (20 g, 36.4 mmol), NaBH ₄ (2.48 g, 65.7 mmol) and TMEDA (8.54 g, 73.5 mmol) ) Pd(dppf)Cl ₂ ·DCM (1.70 g, 2.08 mmol) was added to the mixture under N ₂ atmosphere. After addition, the reaction mixture was stirred at 25°C for 14 hours. The reaction mixture was filtered, the filtrate was concentrated, and the residue was purified by FCC on silica gel (EtOAc) to give the title intermediate (15 g, 93% purity, 74% yield) as a brown solid.

Synthetic Method B for Intermediate 2:

tert -Butyl 6-(3-chloro-6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5 in MeOH (100 mL) -1)-2,6-diazaspiro[3.4]octane-2-carboxylate (intermediate 14 ) (22.0 g, 40.1 mmol), Pd/C (wet, 5.0 g, 10%) was added and the resulting mixture was stirred at 25°C for 8 hours under H ₂ atmosphere (30 psi). The reaction mixture was filtered through a pad of Celite and the filtrate was concentrated in vacuo to give the title intermediate (25.0 g, crude), which was used in the next step without further purification.

Intermediate 3 - 2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)-N-ethyl-5-fluo Preparation of rho-N-isopropylbenzamide

tert -Butyl 6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5-yl)- in DCM (5 mL) TFA (0.5 mL, 6.4 mmol) was added to a solution of 2,6-diazaspiro[3.4]octane-2-carboxylate (Intermediate 2 ) (300 mg, 0.583 mmol), and the resulting mixture was incubated at room temperature for 3 days. Stirred for an hour. Then, 10% NaOH (5 mL) solution was slowly added 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 layers were 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.

Compound 61 - tert-butyl (4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5-yl) Preparation of -2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate

2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy) -N -ethyl- in MeOH (15 mL) 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) 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 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC using _Waters % ammonia) from 45% to 75% v/v) to give the title compound (700 mg, 46% yield) as a colorless oil.

Compounds 62 and 63 - tert-Butyl (R)-(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-tri azin-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 production

tert -Butyl (4-(6-(6-(2-(ethyl (isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5-yl)-2, 6-Diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate (Compound 61 ) (200 mg, 0.319 mmol) was purified by SFC on DAICEL CHIRALPAK IG (column: 250×30 mm 10 um; Purification by isocratic elution: EtOH (containing 25% ammonia at 0.1%): supercritical CO ₂ , 40%: 60% (v/v)) gave the title compound (Compound 62 ) (85 mg, 42% yield) and (Compound 63 ) (80 mg, 40% yield) was obtained as a light yellow oil.

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

tert -Butyl ( R )-(4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4- in DCM (4 mL) Triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)carbamate (Compound 62 ) (550 mg, 0.876 mmol) was added to a solution of TFA (4) mL) was added slowly, and the resulting mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted in DCM (40 mL) and the pH value was adjusted to about 12 with aqueous NaOH (2 M, 16 mL) solution. The aqueous layer was extracted with DCM (10 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 in the next step without further purification.

Compound 11 - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)amino)-2-methylhexane-3 -yl)-2,6-diazaspiro[3.4]octane-6-yl)-1,2,4-triazine-6-yl)oxy)benzamide

( R )-2-((5-(2-(6-amino-2-methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl) in DMF (1 mL) -1,2,4-triazin-6-yl)oxy)- N -ethyl-5-fluoro- N -isopropylbenzamide (Compound 64 ) (120 mg, crude), 1-bromo-2 -A mixture of methoxyethane (32 mg, 0.23 mmol), Cs ₂ CO ₃ (222 mg, 0.681 mmol), and NaI (102 mg, 0.680 mmol) was stirred at 80°C for 1 hour through 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 subjected to HPLC on Phenomenex Gemini-NX (column: 150×30 mm). 5 μm; eluent: ACN/H ₂ O (10mM NH ₄ HCO ₃ ) from 51% to 71% (v/v)) and further purified by SFC on 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, found m/z 586.3 [M+H] ⁺ .

Compound A - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl) (methyl)amino)-2-methyl Hexan-3-yl)-2,6-diazaspiro[3.4]octane-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

( R ) -N -ethyl-5-fluoro- N -isopropyl-2-((5-(2-(6-((2-methoxyethyl)amino)-2- in anhydrous MeOH (2 mL) Methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound 11 ) (40.0 mg, A mixture of 0.068 mmol), formaldehyde (55.4 mg, 0.683 mol, 37% in water) and AcOH (8.2 mg, 0.137 mmol) was stirred at 45°C for 1 hour. 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 hour. After cooling to room temperature, the reaction mixture was treated with saturated aqueous NaHCO ₃ (40 mL) to adjust the pH value to about 8 and further extracted with DCM (20 mL × 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain the crude, which was subjected to preparative HPLC on Boston Prime (column: C18 150×30 mm 5um, mobile phase A: H ₂ O (0.04% Ammonia + 10mM NH ₄ HCO ₃ ), mobile phase B: ACN, flow rate: 25 mL/min, gradient conditions B/A 50% to 80% (50% B to 80% B) to obtain the title compound ( 9.62 mg, 99.10% purity, 23.3% yield) was obtained as a yellow oil.

Example 2 - ( R ) -N -ethyl-5- fluoro - N - isopropyl -2-((5-(2-(6-((2- methoxyethyl )(methyl)amino)-2- Synthesis of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method B

Intermediate 7 - Preparation of 4-((tert-butoxycarbonyl)(methyl)amino)butanoic acid

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 hours. 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), Na Dried over ₂ SO ₄ , filtered and concentrated to give the title intermediate (1.80 g, crude) as a colorless oil.

Intermediate 8 - Preparation of tert-butyl (4-(methoxy(methyl)amino)-4-oxobutyl)(methyl) carbamate

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) were added. EDCI (2.23 g, 11.6 mmol) was then added and the reaction mixture was stirred at room temperature for 4 hours. 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 and filtered. and concentrated under vacuum to give the title intermediate (1.70 g, crude) as a colorless oil.

Intermediate 9 - Preparation of tert-butyl methyl (5-methyl-4-oxohexyl) carbamate

of tert -butyl (4-(methoxy(methyl)amino)-4-oxobutyl)(methyl)carbamate ( Intermediate 8 ) (200 mg, crude) in THF (5 mL) cooled at -70°C. Isopropyllithium (3.2 mL, 2.24 mmol, 0.7M in pentane) was added dropwise to the solution under N ₂ atmosphere. The resulting mixture was stirred at -70°C for 2 hours. The mixture was quenched with saturated aqueous NH ₄ Cl (15 mL) and extracted with EtOAc (30 mL x 2). 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. The crude product was further purified by FCC (PE/EtOAc = 10:1) to give the title intermediate (60 mg) as a colorless oil.

Compound 60 - tert-butyl (4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5-yl) Preparation of -2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl) (methyl) carbamate

2-((5-(2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy) -N -ethyl- in MeOH (50 mL) 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) ) ZnCl ₂ (789 mg, 5.79 mmol) was added to the solution. The resulting mixture was stirred at 80°C for 2 hours. 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 washed with DCM (50 mL × 3). Extracted. 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 purified by FCC (DCM/MeOH = 10:1). Further purification gave 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-diazas Pyro[3.4]octane-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide hydrochloride

tert -Butyl (4-(6-(6-(2-(ethyl(isopropyl)carbamoyl)-4-fluorophenoxy)-1,2,4-triazine-5 in DCM (10 mL) -yl)-2,6-diazaspiro[3.4]octan-2-yl)-5-methylhexyl)(methyl)carbamate (Compound 60 ) (1 g, 1.56 mmol) in 4M HCl in dioxane. (5 mL, 20 mmol) was added, and the resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo to give the title compound (960 mg, crude, HCl salt), which was used 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-methyl Hexan-3-yl)-2,6-diazaspiro[3.4]octane-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

N -ethyl-5-fluoro- N -isopropyl-2-((5-(2-(2-methyl-6-(methylamino) hexan-3-yl)-2,6 in DMF (5 mL) -Diazspiro[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) 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 x 3), dried over Na ₂ SO ₄ , filtered and concentrated to give the 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)benz The amide (Compound 68 ) (250 mg, 48% yield) was obtained 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, several batches obtained by method B) SFC using DAICEL CHIRALPAK IG (column: 250×30mm 10um; mobile phase: A: supercritical CO ₂ , B: EtOH (0.1% ammonia), A:B=40:60, 60 mL/min) Preparative HPLC using Boston Prime (column: 150×30mm 5um, mobile phase A: H ₂ O (10mM NH ₄ HCO ₃ ), mobile phase B: ACN, flow rate: 25 mL/min, gradient condition 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 obscured by solvent peaks and are not reported.

LCMS (ESI) (Method 2): R _t = 1.965 min, found m/z 600.3 [M+H] ⁺ .

SFC (Method 11) : R _t = 4.904 min.

Example 3 - ( R ) -N -ethyl-5-fluoro- N -isopropyl-2-((5-(2-(6-((2-methoxyethyl) (methyl)amino)-2- Synthesis of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide (Compound A) - Preparation Method C

Intermediate 227 - tert-Butyl (R)-(1-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-3-methylbutan-2-yl)carba Manufacture of Mate

Boc-L-valine (44.9 kg), 2,2-dimethyl-1,3-dioxane-4,6-dione (32.9 kg) and DMAP in DCM (607 kg) pre-cooled at -10 to 0°C. (35.5 kg) was added over 3 hours to a solution of DCC (55.5 kg) in DCM (613 kg) and aged at -10 to 0° C. for 16 hours. A 10% aqueous citric acid solution (449 kg) was added while maintaining the temperature below 10°C. The resulting slurry was aged at 0-10° C. for 2 hours and then filtered. Washed with filter cake DCM (91 kg). The filtrate was separated and the organic layer was washed with 10% aqueous citric acid solution (2 times 450 kg) and 10% aqueous NaCl solution (449 kg). To the organic phase (1200 kg), acetic acid (75.0 kg) was added maintaining the temperature between -10 and 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 for a further 16 hours at -10 to 0°C. The mixture was warmed to 15-25° C. and aged for 2 hours. The mixture was then washed with 14% aqueous NaCl solution (450 kg), followed by a second wash with 14% aqueous NaCl solution (432 kg) and a final water wash (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 hours, 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.

Intermediate 228 - Preparation of tert-butyl(R)-2-isopropyl-5-oxopyrrolidine-1-carboxylate

tert -Butyl ( R )-(1-(2,2-dimethyl-4,6-dioxo-1,3-dioxan-5-yl)-3-methylbutan-2-yl in toluene (333 kg) ) Carbamate (Intermediate 227 ) (46.7 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 filtrates were concentrated to dryness under reduced pressure to obtain the target compound (31.05 kg, 96% yield) as an oil, which was used directly without further purification.

Intermediate 229 - Preparation of tert-butyl(5R)-2-hydroxy-5-isopropylpyrrolidine-1-carboxylate

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 (1M, 45.2 kg, 54.4 mol) in 2-MeTHF 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 a further 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 solution (170 kg) and water (155 kg). During water washing, the formed emulsion was separated by addition of solid NaCl (3.1 kg). After removing the aqueous layer, the organic layer was concentrated to dryness under reduced pressure to obtain the desired compound (28.5 kg, 91% yield) as an oil, which was used directly without further purification.

Intermediate 230 - Preparation of tert-butyl(R)-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)carbamate

tert -Butyl (5 R )-2-hydroxy-5-isopropylpyrrolidine-1-carboxylate (Intermediate 229 ) (28.55 kg) in DCM (344 kg) was reacted with 2-methyl at 15-25°C. Treated with toxy- N -methylethane-1-amine (12.3 kg, 138.0 mol) and the resulting mixture was aged for 1 hour. Sodium triacetoxyborohydride (40.12 kg) was added in portions over 5 hours while maintaining the temperature between 15 and 25° C., and the resulting mixture was aged for 48 hours. The reaction mixture was quenched by adding 8% aqueous NaOH solution (184 kg) over 2 hours while maintaining the temperature between 15 and 25° C. and the mixture was aged for a further 2 hours. 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 obtain the title intermediate (33.26 kg, 88% yield) as an oil, which was used directly without further purification.

Intermediate 231 - (R)-N ^One -(2-methoxyethyl)-N ^One Preparation of 5-dimethylhexane-1,4-diamine, dihydrochloride

tert -butyl ( R )-(6-((2-methoxyethyl)(methyl)amino)-2- in iso-propanol (25.6 kg) at ambient temperature in a solution of 4 molar HCl in iso-propanol (84.80 kg). Methylhexan-3-yl)carbamate (Intermediate 230 ) (32.38 kg) was added over 3 hours and the mixture was aged for an additional 19 hours at ambient temperature. 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 give the title compound (23.92 kg, 81% yield) as a white solid.

Intermediate 232 - Preparation of ethyl 1-benzyl-3-(chloromethyl)pyrrolidine-3-carboxylate

n -BuLi (2.33 kg, 2.5 M in hexanes, 1.0 equiv) in a solution of DIPEA (952 g, 1.1 equiv) in THF (6 L) cooled to -35 to -25 °C while maintaining the temperature below -25 °C. was added. The resulting mixture was aged at -35 to -25°C for a further 30 minutes and then cooled to between -78 and 60°C. A solution of ethyl 1-benzylpyrrolidine-3-carboxylate (2 kg, 1.0 equiv) in THF (2 L) at -78 to -60°C was added and stirred for an additional 30 minutes. 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 hours. The reaction mixture was added to an aqueous citric acid solution (660 g in 6 LH ₂ O) at a temperature between 0 and 10° C. and the resulting mixture was aged at 20 to 30° C. for a further 20 minutes. After separation of the layers, the aqueous layer was extracted with EtOAc (6 L) and 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 hours, 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 equiv) and the resulting mixture was aged at 20-30° C. for 20 minutes. After layer separation, 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 (46 wt % assayed, giving an analytical yield of 80%).

Intermediate 233 - Preparation of 1-benzyl-3-(chloromethyl)pyrrolidine-3-carbaldehyde

Reaction performed 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) at 26.7 mL/min. Pumped and cooled to -60°C. After cooling, it was then mixed with a cooled solution of DIBAL-H (28.1 mol) in toluene (28 L) at -60°C at a pumping rate of 32.1 mL/min. The mixture was passed through a perfluoroalkoxy (PFA) coil tube reactor at -60°C (total flow rate 58.8 mL/min, residence time 5 seconds). The resulting mixture was mixed with cooled MeOH (-60°C), which was pumped at a rate of 15.2 mL/min. This mixed solution was pumped to another PFA coil tube reactor at -60°C (total flow rate 74 mL/min, residence time 5 seconds). The resulting mixture was collected into a receiver containing 20% by weight of aqueous Rochelle salt (20 V). The layers were separated and the organic phase was washed twice with brine (2 x 44 mL). 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% by weight assayed by HPLC, giving an analytical yield of 85%), which It was used directly without further purification.

¹ H NMR (300 MHz, chloroform-d) : δ 9.62 (s, 1H), 7.39 - 7.20 (m, 5H), 3.83 - 3.57 (m, 4H), 2.96 (d, J = 10.2 Hz, 1H), 2.80 - 2.55 (m, 3H), 2.17 (ddd, J = 13.9, 7.9, 6.1 Hz, 1H), 1.83 (ddd, J = 13.4, 7.8, 5.5 Hz, 1H).

Intermediate 234 - (R)-4-(6-benzyl-2,6-diazaspiro[3.4]octan-2-yl)-N-(2-methoxy ethyl)-N,5-dimethylhexane-1 -Manufacture of amines

1-Benzyl-3-(chloromethyl)pyrrolidine-3-carbaldehyde (Intermediate 233 ) (3.0 kg, 10 wt%) and ( R ) -N ¹ -( 2-methoxyethyl) -N ^1,5 -dimethylhexane-1,4-diamine, dihydrochloride (Intermediate 231 ) (3.47 kg) was added to a solution of triethylamine (2.55 kg, 25.2 mol) at 20 to 30% Added at ℃. The resulting mixture was aged at 20-30° C. for 2 hours. 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 solution (25 L, approximately 16.75 equivalents) was added while maintaining the temperature below 35°C. The resulting mixture was aged at 20 to 30° C. for 25 minutes and the layers were separated. The organic layer was washed with 15 wt % aqueous NaCl (10 L), the layers were separated again and water (18 L) was charged to the organic phase. The pH of the aqueous phase was adjusted to 6-7 with 4M aqueous HCl while maintaining the internal temperature below 35°C. The organic phase was then discarded and the aqueous phase was separated and basified with K ₂ HPO ₄ to pH 8-9.

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 streams were washed three times (3 x 40 L) with methyl tert-butyl ether. To the resulting aqueous layer was added additional methyl tert -butyl ether (83 L) and the aqueous phase was basified to pH 9-10 with 8 wt % aqueous NaOH while maintaining the temperature between 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, followed by flushing with methanol three times (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 purified further. I used it right away.

Preparation of Intermediate 234a (Citric Acid Salt of Intermediate 234)

EtOH (80 ml) and intermediate 234 (20 g) were added in a round bottom flask. Next, a solution of citric acid in 0.5 M EtOH (100 ml; 1 equiv) was added to the mixture at room temperature in a round bottom flask. Subsequently, the mixture was evaporated to dryness (Rotavap, 40° C.). Acetonitrile (200 ml) was added to the residue and the mixture was evaporated to dryness (Rotavap, 40° C.). Acetonitrile (100 ml) was added to the residue and stirred at room temperature on a magnetic heating plate overnight. Finally, intermediate 234a was filtered and dried at room temperature.

Preparation of the crystalline form of the citrate salt of Intermediate 234 (Intermediate 234b)

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 became homogeneous (approximately 10 minutes). Next, the mixture was cooled to 50°C at a rate of 0.5°C/min. Next, seeds were added (19 mg of intermediate 234a; 0.5 w/w %) and the mixture was aged with stirring for 3 hours and 30 minutes. Next, the mixture was cooled nonlinearly to 20°C over 8 hours at a factor of 2,3. The resulting mixture was stirred overnight and the product was filtered and dried (overnight at room temperature in the hood). After isolation, intermediate 234b was obtained as a crystalline form of the citric acid salt of intermediate 234 (2.75 g; yield 73.9%). The obtained ratio of intermediate/citric acid is 3/2 (NMR).

The nonlinear cooling mentioned above was carried out according to the following equation:

A new linear ramp starts every 30 seconds during the cool down period. The ramp is calculated according to the formula:

T _set : set value for each new lamp

T _{start value} : mixture temperature measured at the start of the cooling trajectory

T _{end value} : defined end value of the cooling trajectory

t _action : actual time from start of cooling

Duration: defined cooling duration

n: exponent

¹ H NMR (400 MHz, MeOH- d ₄ ) δ ppm 0.91 (3 H, d, J =6.88 Hz) 0.98 (3 H, d, J =6.88 Hz) 1.46 - 1.57 (2 H, m) 1.67 - 1.87 (2 H, m) 1.94 - 2.03 (1 H, m) 2.20 - 2.29 (2 H, m) 2.62 - 2.69 (2 H, m) 2.72 - 2.77 (4 H, m) 2.77 - 2.82 (2 H, m) ) 2.90 (2 H, t, J =7.32 Hz) 2.95 - 3.02 (2 H, m) 3.07 - 3.16 (2 H, m) 3.16 - 3.22 (2 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).

Intermediate 224 - Preparation of (R)-N-(2-methoxyethyl)-N,5-dimethyl-4-(2,6-diazaspiro [3.4]octan-2-yl)hexan-1-amine

Methanesulfonic acid (MSA) (11 kg), ( R )-4-(6-benzyl-2,6-dialyte) to palladium hydroxide on carbon (1.2 kg) in EtOH (1.47 kg) cooled to -5 to 5°C. Jaspiro[3.4]octan-2-yl- N- (2-methoxyethyl) -N ,5-dimethylhexan-1-amine (Intermediate 234) (10 kg) and EtOH (250 L) were added to the mixture. was warmed to 35-45° C. and stirred under hydrogen atmosphere (0.27-0.40 MPa) for 16-20 hours.The mixture was filtered over diatomaceous earth (20 kg) and the pad was washed with EtOH (24 L).The filtrate was filtered under reduced pressure. (<40°C) concentrated to 2-3 volumes, followed by two flushes with 2-MeTHF (73 kg and 47 kg) to give a 2-3 volume solution, diluted with 2-MeTHF (65 kg), 10 % aqueous sodium sulfate (30 kg) was added and the mixture was cooled to 0 to 10° C., then 16% aqueous NaOH solution (50 kg) was added to adjust the pH to 13 to 14. The temperature was adjusted to 15 to 25° C. Stirred for 30-60 minutes. 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 ( After concentrating to 3-4 volumes under reduced pressure (<40°C), the resulting solution was diluted with 2-MeTHF (30 kg) and dried by passing through 4A molecular sieves (25 kg). Washed with 2-MeTHF (30 kg) The final solution was concentrated to give the desired compound (6.7 kg) in 79% corrected yield as an oil with an analytical purity of 90.1%.

Intermediate 225 - (R)-4-(6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro [3.4]octan-2-yl) Preparation of -N-(2-methoxyethyl)-N,5-dimethylhexan-1-amine

( R ) -N- (2-methoxyethyl) -N ,5-dimethyl-4-(2,6-diazaspiro[3.4]octan-2-yl)hexan-1-amine (Intermediate 224)( 100 g) were 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% NaHCO ₃ aqueous solution was added, and the mixture was warmed to 20 to 30°C and stirred for 30 to 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 by passing through a 4Å molecular sieve (220 g) and washed with 2-MeTHF (180 g). The title intermediate was provided as a 14.8 wt% solution in 2-MeTHF in 90% analytical 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

Synthetic Method A for Compound 393:

N -ethyl-5-fluoro-2-hydroxy- N -isopropylbenzamide (Intermediate 28) (1.10 g, 4.88 mmol), ( R )-4-(6-(3) in anhydrous THF (15 mL) ,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazaspiro[3.4]octan-2-yl)- N -(2-methoxyethyl)- N ,5 A mixture of -dimethylhexan-1-amine (Intermediate 225) (1.70 g, 3.82 mmol) and DBU (750 mg, 4.93 mmol) 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 FCC (MeOH/DCM = 0% to 10%) to give a yellow oil (1.40 g). Obtained, this was SFC on DAICEL CHIRALPAK AD (column: 250×50 mm, 10 um; mobile phase: A: supercritical CO ₂ , B: EtOH (0.1% ammonia), A:B = 50:50, 70 mL/min ; Column temperature: 38°C; Nozzle pressure: 100Bar; Nozzle temperature: 60°C; Evaporator temperature: 20°C; Trimmer temperature: 25°C; Wavelength: 220nm) to obtain the title compound (1.0 g).

Synthetic Method A for Compound 393:

( R )-4-(6-(3,6-dichloro-1,2,4-triazin-5-yl)-2,6-diazospiro[3.4]octane in 2-MeTHF (40 g) -2-yl)- N -(2-methoxyethyl)- N ,5-dimethylhexan-1-amine (Intermediate 225) (14.8 wt% solution in 676 g of 2-MeTHF, 100 g of calibrated Intermediate 225) And to a 2-MeTHF solution of N -ethyl-5-fluoro-2-hydroxy- N -isopropylbenzamide (Intermediate 28) (50.6 g) was added tetramethylguanidine (31 g) at 20 to 30° C. The mixture was stirred for 40 to 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 solution (2 x 500 g) and once with 10% aqueous Na ₂ SO ₄ solution (500 g). The organic layer was concentrated to 2.2 to 3.0 volumes under reduced pressure (<40°C) and flushed three times with MeOH (1 Obtained as a 60.1 wt% solution in 86% analytical yield.

Compound A - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl) (methyl) amino)-2-methyl Hexan-3-yl)-2,6-diazaspiro[3.4]octane-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide

( R )-2-((3-Chloro-5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexan-3-yl)-2,6-diazas Pyro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy- N -ethyl-5-fluoro- N -isopropylbenzamide (Compound 393) (163.93 g of MeOH A methanol solution of 100 g of calibrated compound 393), palladium on carbon (10 g) and MeOH (316 g) was stirred at 20 to 30° C. under a hydrogen atmosphere (0.20 to 0.30 Mpa) for 18 hours. The mixture was stirred over 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 triturated with isopropyl acetate (IPAc, 870 g). Concentrated to about 3 volumes by flushing. The mixture was then diluted with IPAc (696 g) and 20% aqueous Na ₂ CO ₃ solution was added (500 g). The mixture was stirred for 30-60 minutes. The aqueous layer was removed. The organic layer was washed with water (500 g) and then concentrated under reduced pressure to about 3 volumes at <45° C. The title intermediate was obtained as a 48.1 wt% solution in IPAc in approximately 90% analytical yield.

Example 4 - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2- Synthesis of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide oxalate (Compound A3)

Compound A3

( R ) -N -ethyl-5-fluoro- N -isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)) in 20 mL of ACN (20 mL) 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) was added to oxalic acid (81.0 mg, 0.900 mmol). After addition, the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was then concentrated, the residue redissolved in ACN and deionized water, and lyophilized to give the title compound (350 mg) as a white solid.

¹ H NMR (400 MHz, methanol-d ₄ ): 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).

¹ H NMR (400 MHz, DMSO-d ₆ ): 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): R _t = 1.969 min, found m/z 600.4 [M+H] ⁺ .

Example 5 - Synthesis of Compound A1

To a solution of Compound A (48 wt% solution in 207.90 g of IPAc, 100 g of Active Compound A) in IPAc (360 g) was added EtOH (63 g) at 20-25°C. The solution was then treated with concentrated HCl (32.9 g) in EtOH (49.5 g) over approximately 15 minutes. The mixture was seeded with crystalline Compound A1 seeds (2 g, 2% seed loading) and then aged for 18 hours. IPAc (870 g) was added slowly over 4 hours at 20-25° C. and the slurry was stirred for an additional 18 hours. 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 slightly crystalline compound A1 as a white solid (91.0% yield, 115.4 g). (Note: The small amount of seed material used in the reaction was obtained through a similar reaction protocol on a smaller scale.)

Recrystallization: A solution of slightly crystalline compound A1 (100 g), EtOH (166 g), purified water (21.5 g) and IPAc (178 g) was stirred at 20-30° C. for 0.5-2 hours to obtain a clear solution. did. Extra 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 stirred slowly at 20-30° C. over 12 hours, and the slurry was stirred for a further 55-60 hours. The product was filtered, washed with IPAc (158 g) and dried in vacuo at 20-30° C. to give Compound A1 as a white solid (85% yield, 85.0 g, net).

¹ HNMR (DMSO-d ₆ , 400 MHz): 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): R _t = 0.669 min, found m/z 600.5 [M+H] ⁺ .

Example 6 - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2- Crystalline form of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate salt hydrate Synthesis of Form A (Compound A4) (equivalent water not determined)

43.06 g benzenesulfonic acid (2 equivalents for free base compound A) was added and dissolved in 840 ml of acetone/water 95/5 v/v mixture. A solution of 192.8 g of Compound A in IPAc (containing 80 g API) was added. The material was dissolved to produce a clear solution. A further 80 ml IPAc was added and the temperature was adjusted to 25°C. 2% of seeds were added and the mixture was stirred at 25°C for 1 hour. 28.8 V (2312 ml) of IPAc was then added over a period of 8 hours. The suspension was then stirred at 25°C for 18 hours. The suspension was filtered and washed with 320 ml of a mixture of acetone/water/IPAc 23.75/1.75/75 v/v/v. 122.91 g of crystalline Form A bis-besylate hydrate (equivalent water not determined) was obtained.

Those skilled in the art will understand that the small amounts of initial seed material used in the above reaction can be obtained through similar reaction protocols on a smaller scale without seed addition and waiting for spontaneous nucleation.

Initial seeds of besylate salts were also obtained during salt screening experiments. In these experiments, 100 mg of free base was weighed into a 2 mL vial, and then 200 μL of ethyl acetate or acetone was added to dissolve the free base. One equivalent of counter ion (benzenesulfonic acid) was added to the sample and the sample was stirred at 25°C for 3 days. The obtained suspension was centrifuged and initial seeds were obtained.

An appropriate amount of ( R )-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methyl Crystalline form of hexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate salt hydrate. A was dissolved in deuterated DMSO and 1D ¹ H 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. did.

¹ H NMR (600 MHz, DMSO- d ₆ ) δ ppm 0.69 (br s, 2 H) 0.82 - 0.98 (m, 9 H) 1.07 (br s, 4 H) 1.31 - 1.46 (m, 1 H) 1.51 ( br d, J =2.91 Hz, 1 H) 1.69 (br d, J =3.45 Hz, 2 H) 1.98 (br s, 1 H) 2.06 - 2.45 (m, 2 H) 2.77 (br s, 3 H) 2.87 - 3.19 (m, 3 H) 3.24 (br s, 1 H) 3.31 (s, 6 H) 3.64 (br s, 4 H) 3.71 - 4.59 (m, 7 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 - (R)-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2- Crystalline form of methylhexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate salt hydrate Alternative synthesis of Form A (Compound A4) (equivalent water not determined)

A mixture of isopropanol/water 95/5 (24 ml) was charged to the flask and heated to 40°C. Benzenesulfonic acid (4.31 g; 98%) was added. A solution of Compound A in IPAc (containing 8 g of Compound A) was then added. Another 16 ml of IPAc was added. Then, 2% of seeds were added and the mixture was stirred at 40° C. for 1 hour. IPAc was then added dropwise (115.2 ml) over a period of 8 hours. Next, the mixture was 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 yield 11.44 g of crystalline Form A bis-besylate hydrate (equivalent water not determined).

Crystalline Form A

( R )-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methylhexane-3 -yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate salt hydrate crystalline form A -Can be characterized by line powder diffraction patterns.

X-ray powder diffraction (XRPD) analysis was performed on a PANalytical Empyrean diffractometer. The instrument is equipped with a Cu-Kα X-ray tube using iCore and dCore tunable optics for the incident and diffracted beams, respectively. Compounds were loaded into the cavity of a 16 mm sample holder using a back loading technique.

Samples were run for XRPD using the following method:

)Tube: Cu: K-alpha (λ=1.541874

)

Generator: Voltage: 45 kV; Current: 40 mA

Geometry: Bragg-Brentano

Scan Mode: Continuous Scan

Scan range: 3 to 35 degrees.

Step size: 0.0131 degrees.

Counting time: 30 seconds

Spinner rotation time: 1 second

Incident beam path (iCore)

Program divergence slit: automatic

Probe length: 10 mm

Solar slit: 0.03 rad

Mask 1: 14 mm

Mask 2: 6 mm

Width: 7.7 mm

Diffraction Beam Path (dCore)

Anti-scatter slit: automatic

Probe length: 10 mm

Solar slit: 0.04 rad

Detector: PIXcel3D- Medipix3 1×1

Those skilled in the art will understand that typically the diffraction pattern and peak positions are substantially independent of the diffractometer used and whether a particular calibration method is used. Typically, peak positions may differ by less than about ±0.2° 2 theta. The intensity (and relative intensity) of each particular diffraction peak may also vary depending on a variety of 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 2theta ± 0.2 degrees 2theta. The X-ray powder diffraction pattern may further include one or more peaks at 13.7, 14.5, 14.7, 15.0, 16.5, 17.8, 19.0, 19.4, 20.1 degrees 2theta ± 0.2 degrees 2theta.

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 the peaks specified in Table 2. .

Form A may be further characterized by an X-ray powder diffraction pattern comprising the peaks specified in Table 2, wherein the relative intensity of the peaks is greater than about 2%, preferably greater than about 5%, more preferably greater than about 10%, more preferably greater than about 15%. However, those skilled in the art will recognize that the relative intensities of peaks may vary between different samples and between different measurements on the same sample.

Form A may also be further characterized by an X-ray powder diffraction pattern substantially as shown in Figure 1.

Table 2 shows ( R )-N-ethyl-5-fluoro-N-isopropyl-2-((5-(2-(6-((2-methoxyethyl)(methyl)amino)-2-methyl Crystalline form of hexan-3-yl)-2,6-diazaspiro[3.4]octan-6-yl)-1,2,4-triazin-6-yl)oxy)benzamide bis-besylate hydrate BSA salt Provides a list of peaks and relative intensities for the XPRD of A (Figure 1).

[Table 2]

Compound A4 described herein is disclosed in PCT/CN2021/100466 (filed June 17, 2021), which is incorporated herein by reference in its entirety for all purposes.

Analysis method

Analytical information for compounds in the table below or above was generated using the analytical methods described below.

NMR-Methods

Some NMR experiments were performed using a Bruker Avance III 400 spectrometer at ambient temperature (298.6 K) with internal deuterium locking and equipped with a BBO 400 MHz S1 5 mm probe head with a z gradient, 400 MHz for protons and 400 MHz for carbon. It was performed operating at 100 MHz. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Some NMR experiments were performed using a Varian 400-MR spectrometer at ambient temperature (298.6 K) with internal deuterium locking and equipped with a Varian 400 4NUC PFG probe head with a z gradient, 400 MHz for protons and 100 MHz for carbon. It was performed operating at MHz. Chemical shifts (δ) are reported in parts per million (ppm). J values are expressed in Hz.

Some NMR experiments were performed using a Varian 400-VNMRS spectrometer at ambient temperature (298.6 K) with internal deuterium locking and equipped with a Varian 400 ASW PFG probe head with a z gradient, 400 MHz for protons and 100 MHz for carbon. It was performed operating at MHz. 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 300 spectrometer at ambient temperature (298.6 K) with internal deuterium locking and equipped with a PA BBO 300S1 BBF-H-D-05 Z 5 mm probe head with a z gradient, for protons. It was performed operating at 300 MHz for carbon 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 procedure

High-performance liquid chromatography (HPLC) measurements were performed using columns and LC pumps, diode-array (DAD) or UV detectors as specified in the respective methods. HPLC details are provided in Table 3 below. If necessary, additional detectors were included (see Tables 3 and 4 below).

Flow from the column was allowed to reach a mass spectrometer (MS) configured with an atmospheric pressure ion source. It is within the knowledge of a person skilled in the art to set tuning parameters (e.g. scanning range, retention time...) to obtain ions that allow identification of the nominal monoisotopic molecular weight (MW) of the compound. Data acquisition was performed using appropriate software.

Compounds are described by their experimental retention time (R _t ) and ion. Unless otherwise specified in the table of data, reported molecular ions correspond to [M+H] ⁺ (protonated molecule) and/or [MH] ^- (deprotonated molecule). If the compound is not directly ionizable, the type of adduct is specified (i.e. [M+NH ₄ ] ⁺ , [M+HCOO] ^- etc...). For molecules with multiple isotopic patterns (Br, Cl), the values reported are those obtained for the lowest isotopic mass. All results were obtained with experimental uncertainties normally associated with the methods used.

Hereinafter, “SQD” refers to single quadrupole detector, “RT” refers to room temperature, “BEH” refers to cross-linked ethylsiloxane/silica hybrid, “HSS” refers to high-strength silica, and “DAD” refers to "refers to a diode array detector.

[Table 3]

SFC for analysis

General procedure for SFC method

SFC measurements are performed using an analytical supercritical fluid chromatography (SFC) system consisting of a diode array detector equipped with a binary pump for delivering carbon dioxide (CO ₂ ) and modifiers, an autosampler, a column oven, and a high-pressure flow cell withstanding up to 400 bar. It was performed using . Analytical SFC details are provided in Table 4 below. If a mass spectrometer (MS) is equipped, flow from the column is allowed to reach the (MS). It is within the knowledge of a person skilled in the art to set tuning parameters (e.g. scanning range, retention time...) to obtain ions that allow identification of the nominal monoisotopic molecular weight (MW) of the compound. Data acquisition was performed using appropriate software.

[Table 4]

Pharmacology part

1) Menin/MLL homogeneous time-resolved fluorescence (HTRF) analysis

In an untreated white 384-well microtiter plate, 40 nL of 200 4 μL of 2X terbium chelate labeled menin (see below for preparation method) was added. After incubation of terbium chelate-labeled menin with test compounds for 30 min at ambient temperature, 4 μL ₂ cyanate) was added, the microtiter plate was centrifuged at 1000 rpm for 1 min and the assay mixture was incubated for 15 min at ambient temperature. The relative amount of menin·FITC-MBM1 complex present in the assay mixture was determined by measuring the terbium/FITC donor/acceptor fluorophore using an EnVision microplate reader (ex. 337 nm/terbium em. 490 nm/FITC em. 520 nm) at ambient temperature. It is determined by measuring the homogeneous time-resolved fluorescence (HTRF) of the pair. The degree of fluorescence resonance energy transfer (HTRF value) is expressed as the ratio of the fluorescence emission intensity of FITC and 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 typically performed using an 11-point, 4-fold serial dilution scheme starting at 10 μM.

Compound efficacy was determined by first calculating the inhibition rate (%) at each compound concentration according to Equation 1.

[Equation 1]

%inhibition = ((HC - LC) - (HTRF ^compound - LC)) / (HC - LC)) *100

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 menin binding, and HTRF ^compound is the HTRF value measured in the presence of the test compound. HC and LC HTRF values represent an average of 10 or more replicates per plate. For each test compound, the % inhibition values were plotted against the logarithm of the test compound concentration and the IC ₅₀ values were derived from fitting these data to Equation 2:

[Equation 2]

% Suppression = Bottom + (Top-Bottom)/(1+10^((log IC ₅₀ -log[cmpd])* h ))

Here, Bottom and Top are the lower and upper asymptotes of the dose-response curve, respectively, IC ₅₀ is the concentration of the compound that inhibits the signal by 50%, and h is the Hill coefficient.

Preparation of terbium cryptate labeling of menin: menin (a.a 1-610-6xhis tag, 2.3 mg/m in 20 mM Hepes (2-[4-(2-hydroxyethyl)-1-piperazinyl]ethane sulfonic acid) mL, 80 mM NaCl, 5 mM dithiothreitol (DTT), pH 7.5) was labeled with terbium cryptate as follows. 200 μg of menin was buffer exchanged with 1 × Hepes buffer. 6.67 μM methine was incubated with an 8-fold molar excess of NHS (N-hydroxysuccinimide)-terbium cryptate for 40 min at room temperature. The reaction was carried out on a NAP5 column using elution buffer (0.1M Hepes, pH 7 + 0.1% BSA (bovine serum albumin)) to purify half of the labeled proteins from the unlabeled ones. The other half was eluted with 0.1M phosphate buffered saline (PBS), pH7. 400 μl of eluate was collected for each, aliquoted and frozen at -80°C. The final concentration of terbium-labeled menin protein was 115 μg/mL in Hepes buffer and 85 μg/mL in PBS buffer.

Menin protein sequence (SEQ ID NO: 1):

MGLKAAQKTLFPLRSIDDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVIPTNVPELTFQPSPAPDPPGGLTYFPVADLSIIALYARFTAQIRGAVDLSLYPREGGVSSRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGTKLDSSGVAFAVVGACQALGLRDVHLALSEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTV NAGVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLWLLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIASAKTYYRDEHIYPYMYLAGYHCRNRNNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVANDVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKWEEGSPT PVLHVGWATFLVQSLGRFEGQVRQKVRIVSREAEAAEAEEPWGEEAREGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPEGGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINSSAIKLQLTAQSQVQMKKQKVSTPSDYTLSFLKRQRKGLHHHHHH

2a) Proliferation assay

The antiproliferative effects of the menin/MLL protein/protein interaction inhibitor test compounds were evaluated in human leukemia cell lines. The cell line MOLM-14 harbors the MLL translocation and expresses the MLL fusion protein MLL-AF9 and the wild-type protein of the second allele, respectively. OCI-AML3 cells carrying NPM1c gene mutations were also tested. MLL rearranged cell lines (e.g. MOLM-14) and NPM1c mutant cell lines display an elevated HOXA/MEIS1 gene expression signature similar to stem cells. 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). It was spread from. During culture, cells were maintained at 0.3 to 2.5 million cells per ml and the number of passages did not exceed 20.

To evaluate the antiproliferative effect, 200 MOLM-14 cells, 200 OCI-AML3 cells, or 300 KO-52 cells were seeded in 96-well round-bottom, ultra-low attachment plates (Costar, Cat. No. 7007). Cell seeding numbers were selected based on the growth curve to ensure linear growth throughout the experiment. Test compounds were added at various concentrations and the DMSO content was normalized to 0.3%. Cells were incubated at 37°C and 5% CO ₂ for 8 days. Spheroid-like growth was measured in real time via live cell imaging (IncuCyteZOOM, Essenbio, 4x objective) with images acquired on day 8. Confluence (%), a measure of spheroid size, was determined using integrated analysis tools.

Confluence of each well was calculated as a measure of spheroid size to determine the effect of the test compound over time. 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 IC ₅₀ values were calculated as the rate of change of confluence as follows:

LC = low control: cells treated, for example, with the cytotoxic agent staurosporine at 1 μM, or cells treated with, for example, a high concentration of an alternative reference compound;

HC = high control: average % confluence (DMSO treated cells);

% Effectiveness = 100 - (100*(Sample-LC)/(HC-LC)); and

IC ₅₀ was calculated using GraphPad Prism (version 7.00). A dose-response equation was used for a plot of % effect versus Log10 compound concentration with variable slope and fixed maximum at 100% and minimum at 0%.

2b) MEIS1 mRNA expression analysis

MEIS1 mRNA expression upon compound treatment was examined by Quantigene Singleplex analysis (Thermo Fisher Scientific). This technique allows direct quantification of mRNA targets using probes that hybridize to defined target sequences of interest, and signals are detected using the multimode plate reader Envision (PerkinElmer). The MOLM-14 cell line was used in this experiment. Cells were plated in 96-well plates at 3,750 cells/well in the presence of increasing concentrations of compounds. After 48 hours of incubation with the 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 well as a blocking probe as normalization controls. Cell lysates were then transferred to custom assay hybridization plates (Thermo Fisher Scientific) and incubated at 55°C for 18 to 22 hours. Subsequently, the plate was washed to remove unbound material, and then the preamplifier, amplifier, and label probe were added sequentially. Signals (=gene number) were measured with a multimode plate reader Envision. IC ₅₀ was calculated through dose-response modeling using appropriate software. For all non-housekeeper genes, responses are 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 change was calculated by dividing the normalized value for the treated sample by the normalized value for the DMSO treated sample. The fold change of each target gene was used to calculate IC ₅₀ .

The results are summarized in Table 5 below.

[Table 5]

3) Mouse PK (in vivo T _1/2 and oral bioavailability)

In vivo pharmacokinetics (PK) were determined by a single intravenous (IV, 0.5 or 1.0 mg/kg dose of 2.5 ml/kg) test article formulated in 20% (w:vol) HP-β-CD solution or pyrogen-depleted water. ) or oral (PO, 5 mg/kg administered as 10 ml solution/kg) and then evaluated in fasted male CD-1 mice (6 to 8 weeks of age).

Plasma and/or whole blood samples were collected from the dorsal metatarsal vein at desired time points via 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 key pharmacokinetic parameters was performed using WinNonlin (PhoenixTM, version 6.1) or similar software.

4) Metabolic stability of human/mouse liver microsomes

Experimental Procedure

The purpose of this study is to determine the in vitro metabolic stability of test compounds in human and mouse liver microsomes and to provide quantitative information on metabolic turnover (i.e., determine the apparent intrinsic clearance of the test).

Test items were prepared at a stock concentration of 10 mM in DMSO. For metabolic turnover measurements, the final working solution was 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).

Incubation was performed as follows. First, liver microsomes were thawed on ice, and a master solution containing liver microsomes was prepared in 100 mM PBS (phosphate-buffered saline) at pH 7.4. Next, the liver microsome solution was added to the incubation plate and 10 mM NADPH (nicotinamide-adenine dinucleotide phosphate) (MW: 833.4 g/mol; Roche Diagnostics GmbH, Germany. Phosphate buffer (100 mmol/L; dissolved in pH 7.4)). The mixture was mixed for 10 seconds and preheated in the incubation plate to 37°C for 10 minutes. The metabolic reaction was initiated by adding 5 μL of a 100 μM working solution for the test compound or positive control compound to the incubation plate (final test article 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 organic solvent proportion of the incubation mixture is 1% and DMSO≤0.02%.

At selected time points, 50 μL of the incubated mixture was transferred to a quenching plate containing 200 μL of cold methanol to quench the reaction. After sampling at all time points, the quenching plate was centrifuged at 4000 rpm for 40 min to precipitate the proteins. A total of 90 μL of supernatant was transferred to an assay plate and ultrapure H ₂ O water was added to each well to perform LC/MS/MS analysis. All incubations and analyzes were performed in duplicate.

data analysis

All calculations were performed using Microsoft Excel. The slope value k was determined by linear regression of the residual percentage of parent drug versus the natural logarithm of the incubation time curve. The results are summarized in Table 6 below.

The in vitro half-life (in vitro t _1/2 ) was determined from the slope values:

In vitro t _1/2 = - (0.693 / k)

The conversion of in vitro t _1/2 (in minutes) to in vitro intrinsic clearance (in vitro CL _int , in μL/min/mg protein) was performed using the equation:

[Table 6]

5) Protocol for pharmacodynamic (PD) activity in subcutaneous (sc or SC) xenografts of MOLM-14 or OCI-AML3 cells

Test agent and control group

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. The dose was adjusted daily depending on the individual's body weight. For each study, working stocks of compound A3 were prepared once per week and stored at room temperature. Compound A3 was administered orally (PO) daily.

analyze

The in vivo pharmacodynamic (PD) activity of the compounds was assessed in subcutaneous (SC) xenografts of MOLM-14 cells or OCI-AML3 cells. Nude NMRI mice bearing MOLM-14 or OCI-AML3 tumors (Crl: NMRI-Foxn1nu/-) were treated with three daily doses of vehicle or compound. Plasma samples were collected 23 hours after the second dose, 0.5 hours after the last dose, and 16 hours after the last dose, and tumor samples were collected 16 hours after the last dose. QuantiGene Plex technology (Thermo Fisher Scientific) was used to investigate the effect of compounds on the expression of several menin-KMT2A target genes (e.g. MEIS1, MEF2C, FLT3 ). Frozen tumors were homogenized, transferred to individual lysis matrix tubes containing lysis buffer, and incubated at 55°C for 30 min. 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 to 22 hours. Subsequently, the plate was transferred to a magnetic separation plate and washed to remove unbound material from the beads, followed by sequential hybridization of the preamplifier, amplifier and label probes and subsequent streptavidin phycoerythrin binding. Signals from beads were measured with a Luminex FlexMap three-dimensional instrument. For all non-housekeeper genes, responses are 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 change was calculated by dividing the normalized value for the treated sample by the normalized value for the DMSO treated sample. The results are summarized in Tables 7 and 8 below.

[Table 7]

[Table 8]

Tables 7a and 8a show median values based on replicate experiments under optimized conditions using fresh tumor samples.

[Table 7a]

[Table 8a]

6) Efficacy study in MOLM-14 subcutaneous model

Test agent and control group

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. The dose was adjusted daily depending on the individual's body weight. For each study, working stocks of compound A3 were prepared once per week and stored at 25°C.

animal

Female NMRI nude mice (MOLM-14 SC) were used when they were approximately 6 to 8 weeks of age and weighed approximately 25 g. All animals were allowed to adapt and recover from transport-related stress for at least 7 days before being used in experiments. 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 before use and replaced weekly. Additional details are provided in Table 9 below.

[Table 9]

Tumor models and cell culture methods

Human AML MOLM-14 cells were cultured in the indicated complete culture medium (RPMI 1640 + 10% HI-FBS + 2mM L-glutamine + 50ug/ml gentamicin) at 37°C in 5% CO ₂ . Cells were harvested during logarithmic growth and resuspended in cold (4°C) Roswell Park Memorial Institute (RPMI) 1640 in serum-free medium.

Each mouse was injected into the right flank with 5 × 10 ⁶ MOLM-14 cells in 50% Matrigel in a total volume of 0.2 mL using a 1cc syringe and 27-gauge needle.

study design

Compound A3 was administered orally (PO) daily.

Day 0 is the date of tumor cell transplantation and study initiation.

Mice bearing SC MOLM-14 tumors were randomized 16 days after tumor implantation and assigned to treatment groups according to tumor volume (mean approximately 130 mm ³ ; n=10/group). Treatment with vehicle or Compound A3 (30 and 100 mg/kg) began on the same day and was administered orally daily for 21 days. Plasma was collected for pharmacokinetic (PK) analysis at 1, 2, 4, 8, and 23 hours after the last dose (n=4 to 5/group/time point).

animal monitoring

SC tumor volume was measured at least 2 to 3 times per week for each animal during the study period.

calculate

Tumor volume was calculated using the formula:

Tumor volume (mm ³ )=(D×d ² /2); Here, 'D' represents the larger diameter of the tumor and 'd' represents the smaller diameter, as determined 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, calculated as follows: %ΔTGI=([(TV _c TVc ₀ )(TV _t TV _t0 )]/(TV _c TVc ₀ )) ×100; Where, ‘TV _c ’ is the average tumor load in the given control group, ‘TV _c0 ’ is the average initial tumor load in the given control group, ‘TV _t ’ is the average tumor load in the treatment group, and ‘TV _t0 ’ is the average tumor load in the treatment group. is the average initial tumor burden. %TGI was defined as the difference between the mean tumor volumes of the treatment and control groups calculated as follows:

%TGI=((TV _c TV _t )/TV _c )×100; Here, ‘TV _c ’ is the average tumor volume of a given control group and ‘TV _t ’ is the average tumor volume of the treatment group. As defined by 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 control group, was calculated as %TR= (1-mean (TV _t i/TV _t0 i)) × 100; , where ‘TV _t i’ is the tumor burden of an individual animal in the treatment group, and ‘TV _t0 i’ is the initial tumor burden of the animal.

data analysis

Tumor volumes were graphically displayed using Prism software (GraphPad Version 7 or 8). Statistical significance for most studies was assessed for the Compound A3 treated group compared to the HPβCD vehicle treated control group on the final day of the study when more than two-thirds 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-Effect (LME) analysis in R software version 3.4.2 (using version 4.0 of the Shiny application developed internally at Janssen), where: Treatment and time were used as fixed effects and animal as a random effect. If individual longitudinal response trajectories were not linear, log transformation was performed.

Information derived from this model was used to conduct pairwise treatment comparisons of tumor volume between control or all treatment groups. The results are shown in Figure 2.

7) Ca ²⁺ -Cardio-electrophysiological effects of test compounds in synchronously beating human pluripotent stem cell-derived cardiomyocytes (hSC-CM) using fluorescence analysis (CTCM human).

protocol

Compounds were tested in 96 well plates.

Compounds were tested in Cor.4U®-cardiomyocytes or iCell® cardiomyocytes2 at 0.1 μM, 0.2 μM, 0.5 μM, 1 μM, 2.5 μM and 5 μM (n = 4 per dose).

Alternatively, the compounds are mostly 0.1 μM, 0.3 μM in iCell® cardiomyocytes2; Tested at 1 μM, 3 μM, 10 μM and 30 μM (n = 4 per dose).

Positive and Negative Controls

Vehicle control: dimethyl sulfoxide (DMSO). Solutions of compounds in DMSO or its solvent (final concentration of 0.1% DMSO; n=8).

Preparation of test articles and controls

The tested compounds were dissolved in DMSO at 1000 times the intended concentration. Compound “seed plates” were created containing test compounds and positive and negative controls at 1000 times the final concentration. On the day of the experiment, these stock solutions were diluted (in round-bottom composite plates) to 2 times the intended concentration with Tyrode (Sigma) supplemented with 10 mM HEPES (Gibco). The final DMSO concentration in the test solution and vehicle control was 0.1%.

cell

hSC-CM (Cor.4U ^® cardiomyocytes) were obtained from CDI (Ncardia, Germany). Cells are pre-plated and seeded in fibronectin-coated 96-well plates at a density suitable to form a monolayer, and culture is maintained in a stage incubator (37°C, 5% CO ₂ ) according to the instructions of the cell supplier.

Second-line hSC-derived cardiomyocytes, called iCell® cardiomyocytes 2, were purchased from FUJIFILM Cellular Dynamics (USA). Experiments using test drugs were performed 5 to 7 days after plating cells onto plates to have viable, beating monolayers of hiPSC-derived cardiomyocytes. Beating monolayers in 96-well plates are typically harvested from two vials (approximately 5 million cells/vial) of frozen iCell® cardiomyocytes2, which will be plated in three 96-well plates (approximately 50K/well).

Before starting the experiment

At least 1 hour before the start of the experiment, normal cell medium was replaced with Tyrode's solution containing calcium dye (see below).

Cal 520 dye (AAT Bioquest) was dissolved in 11 ml of Tyrode supplemented with 10 mM HEPES and preheated to 37°C before addition 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 plates were incubated at 37°C/5% CO ₂ for 45 minutes. Cells were incubated at 37°C for 5 minutes.

Experiment

Spontaneous electrical activity is recorded using Cal520™ (AAT Bioquest) calcium fluorescent dye signaling. This dye integrates total intracellular calcium activity throughout the well. Dissolve one bottle of Cal520 dye (50 μg, MW: 1103/mol) in 50 μl of DMSO, a 0.9 mM stock solution. 50 μL of dye stock solution was added to 10 ml of Tryodes solution to achieve a dye concentration of 4.5 μM. Then, 35 μl of this dye solution was added to each well so that the final dye concentration was 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 the Functional Drug Screen System (FDSS/μCell, Hamamatsu, Japan), and recordings were then analyzed offline using appropriate software such as Notocord.

For test runs, cell plates were loaded into FDSS/μCell: Ca ²⁺ transients were measured for 4 min to confirm synchronous beating of cardiomyocytes in each well. All 96 wells were measured simultaneously (sampling interval: 0.06 s, 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 pre-established criteria to ensure synchronous beating in all 96 wells at baseline were excluded from the study and not treated with compound):

T = 0: control period (-5 min to -1 min) + compound addition, then for 3 min

T = 30: measured 29 to 34 minutes after compound addition

During the compound addition step, 100 μl of each double concentrated test solution was simultaneously pipetted into each well.

Data were analyzed offline using appropriate software such as Notocord-Hem (version 4.3).

The following parameters of Ca ²⁺ transient morphology were measured:

- Bit rate (BR)

- amplitude of Ca ²⁺ transients (Amp),

- CTD ₉₀ : Ca ²⁺ transient duration at 90% (time to reach 90% of initial baseline).

The presence of various ‘arrhythmia-like’ activities was also noted during the experiment. These included:

'초기 탈분극 유사'(EAD 유사) 이벤트("과도의 초기 피크 이후 과도 파형의 매우 작은 피크"로 정의됨),

' Initial depolarization ' pseudo ' (EAD-like) events (defined as "a very small peak in the transient waveform after the initial peak of the transient");

‘ Ventricular tachycardia -like ’ (VT-like) events (defined as very fast beat rates); or

' Ventricular fibrillation 'pseudo ' (VF-like) events (defined as "small ^- amplitude, high-velocity Ca ²⁺ waves with irregular, unmeasurable transient potentials)

' Stop beating' of the cell (no ^{Ca2 +} transients observed).

If compound-induced changes in calcium transient signals could not be analyzed by the software, these signals were identified as Below Quality Analysis Level (BQL).

data analysis

The measured data from FDSS-μCell were copied for offline analysis and analyzed and uploaded to the Operations Management System (SPEC-II) for further analysis. Variable values before and after compound administration were collected and transferred to an Excel workbook.

All values (actual units and percentage change from default) are expressed as median (minimum and maximum). The Wilcoxon-Mann-Whitney test was used to compare the changes observed in compound groups relative to their respective baseline values (in real units) with the changes in the solvent control group. Two-tailed tests using Bonferroni correction were performed to adjust for multiplicity. Since there are 10 treatment groups each compared to the solvent group, an alpha level of 0.05/10 (0.005) is considered to reflect a statistically significant difference from the solvent group. All statistical analyzes were performed using appropriate software, such as R software version 3.5.2.

Quality control of hiPSC-CMs on plates:

Plates were rejected if they did not meet the following criteria:

- a steady, regular beat

- Amplitude > 500 relative units

- Beat rate between 25 and 80 beats per minute

- CTD ₉₀ between 300 and 800 ms.

In this study, the hiPSC-CMs on the plates met the above criteria.

These parameters combined with arrhythmia incidence or beat interruption were used to calculate the potential risk level using a weighted scoring method (based on Kopljar et al., Stem Cell Reports 2018. 11, 1365-1377). This risk score is calculated for each concentration by adding weights based on the change in CTD ₉₀ , heart rate and amplitude (ΔΔ%), and tolerance interval (TI) for the incidence of beat arrest and early postdepolarization (EAD). As a result, one of four risk levels will be generated for each concentration. This will be done 30 minutes after incubation with the compounds. The risk levels are:

No risk: Small changes within or unrelated to the vehicle effect level.

Low risk: associated benefits but potentially lower heart disease risk.

High risk: Relatively high risk of heart disease.

Very High Risk: Very high risk of an arrhythmia-like event (EAD).

The 'Risk Score' result provides identification of potential acute cardiac drug-induced effects in free drug equivalents (no plasma proteins added to the well). Assessment of risk identification is conducted using the 'scoring reference book' called CTCM_Scoring_version 1 (Kopljar et al., Stem Cell Reports 2018. 11: 1365-1377], and the level is determined according to the color scheme in Table 10 below. displayed.

[Table 10]

The ranking of test compounds according to risk score severity for Ca ²⁺ transient analysis is determined in HiPSc-CM listed above in various colors and in the relevant table.

result

Using iCell® cardiomyocytes 2 as a cell line

Positive and Negative Controls: Both positive and negative controls showed the expected pharmacological effects in this assay. The results are shown in Tables 11 and 12 below.

[Table 11]

[Table 12]

For Compound A1: In the mouse xenograft model at 30 mg/kg, CTCM human concentration versus free C _max will 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) Membrane potassium current I in hERG transfected cell lines _Kr Impact on

Protocol 1:

Abbreviation

method

Experiments were performed using CHO cells stably expressing the hERG potassium channel. Cells were grown at 37°C and 5% CO ₂ in culture flasks in Ham's F12 medium supplemented with 10% heat-inactivated fetal calf serum, hygromycin B (100 μg/ml), and geneticin (100 μg/ml). I ordered it. Cells were harvested to obtain a cell suspension of single cells for use in the automated patch-clamp system QPatch (Sophion).

Solution: Bath solution contains (in mM) 145 NaCl, 4 KCl, 10 glucose, 10 HEPES ((4-(2-hydroxyethyl)-1-piperazinethanesulfonic acid), 2 CaCl ₂ and 1 MgCl ₂ ( (pH 7.4 with NaOH). The pipette solution contained (in mM) 120 KCl, 10 EGTA (ethylene glycol- bis (2-aminoethylether)-N , N , N ', N' -tetraacetic acid). , 10 HEPES, 5.374 CaCl ₂ and 1.75 MgCl ₂ (pH 7.2 with KOH).

Patch-clamp experiments were performed in voltage-clamp mode and whole-cell currents were recorded by automated patch-clamp analysis utilizing the QPatch system (Sophion). Current signals were amplified, digitized, stored, and analyzed using QPatch analysis software.

The holding potential was -80 mV. hERG current (K ⁺ -selective outward current) was determined as the maximum tail current at -40 mV after 2 s depolarization to +60 mV. The pulse cycling rate was 15 seconds. Short pulses (90 ms) to -40 mV were used as a baseline step to calculate tail current amplitude. After establishing the overall cell composition and stability period, a solvent control (0.3% DMSO) was applied for 5 min, followed by test substances at 3 × 10 ^-7 M, 3 × 10 ^-6 M, 10 ^-5 M and 3 × 10 It was applied in four increasing concentrations of ^-5 M. Each concentration of test substance was applied twice. The effect of each concentration was determined as the average current of three sequential voltage pulses after 5 min. Residual currents were compared to vehicle pretreatment to determine the degree of blocking.

Concentration/response relationships were calculated by nonlinear least squares for individual data points. The half maximal inhibitory concentration (IC50) was calculated through a fitting routine.

Protocol 2:

cell

Compounds, vehicle control and positive control were tested in hERG-transfected HEK293 cells. A human embryonic kidney cell line (HEK293) stably transfected with hERG (Zhou Z et al. Biophysical Journal 1998. 74, 230-241; McDonald TVet al, Nature 1997. 388, 289-292]) was used ( University of Wisconsin, Madison, USA). T175 flasks were used to maintain cell cultures in MEM (Minimum Essential Nutrient Medium, Gibco) supplemented with the following (indicated amounts added to 500 ml MEM): 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% CO ₂ atmosphere (in air).

Cell harvest for analysis

Cells were harvested as described below using accumax™ (Sigma) as the dissociation reagent. The cells were then resuspended in a mixture of 33% DMEM/F12 (Dulbecco's modified Aigle's medium/nutrient mixture F-12-Sigma) medium/67% extracellular physiological solution.

The flask was carefully washed twice with approximately 5 to 10 ml phosphate buffered saline (PBS) (Gibco™) containing 2 mM ethylenediaminetetraacetic acid (EDTA) (Sigma). Cells were separated using approximately 3 ml of accumax™ (cell separation solution) and incubated at 37°C for approximately 5 to 10 minutes. Cold external physiological solution (2-5 ml) was added and the flask was incubated for 5-10 minutes at approximately 4°C. The cell suspension from each flask was then gently dissociated with a 5 ml pipette. The cell suspension was transferred to a low-binding Petri dish (approximately 10 mm in diameter). Each flask was additionally washed with approximately 5 ml of cold external physiological solution and this solution was also added to the Petri dish. The Petri dish was then incubated at approximately 4°C for an additional 5 to 10 minutes. After gently dissociating the cell suspension from the Petri dish again, the cells were transferred to a reservoir kept on an orbital shaker at 16°C and a speed of 200 rpm. Cells were harvested for approximately 20 min before performing the experiment.

compound

A 10 mM solution of the compound was used and plated in a 384 well plate. Aliquots of the stock solution are diluted into recording solution using automated liquid handling (Biomek FXP; final DMSO concentration: 0.03-0.3%). A standard range of screening concentrations ranging from 1 μM to 30 μM was used.

A positive control (E-4031) was included in each run to assess the sensitivity of the assay. Table 13 summarizes the external and intracellular solutions used in the experiment. In Table 13, the composition of the intracellular and extracellular buffers is shown in [mM], and “NMDG” means N-methyl-D-glucamine.

[Table 13]

study design

Using whole-cell patch clamp techniques on transfected cells, ion channels can be studied 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 384PE (Obergrussberger et al, Journal of Laboratory Automation 2016. 21 (6), 779-793). All cells were recorded in whole-cell mode of the patch clamp technique. This module is integrated into the Biomek FXP, a liquid handling pipetting robotic system, for application of cells and compounds, vehicle controls and positive controls.

Various concentrations of the compounds were applied with two cumulative increasing concentrations for the compounds (1 μM and 10 μM, 3 μM and 30 μM, respectively). hERG current was determined as the maximum tail current at approximately 30 mV, and inhibition rates were reported for addition of compound or vehicle and positive control.

After attaching cells to individual holes of the recording chip using a chip filling solution, sealing is increased with a sealing strengthening solution (increasing [Ca ²⁺ ]); Cells were then washed twice with recording solution before using the pressure protocol to enter whole-cell mode.

After whole-cell mode was achieved, test pulses were given for approximately 10 min to quantify hERG currents in control conditions. During this control period, vehicle control solution (recording solution containing 0.03% DMSO) was added to individual wells three times. While continuing the pulse protocol, cumulatively increasing concentrations of vehicle control, compound, or positive control were added. The effects of vehicle, compound and positive control were measured 5 minutes after drug application. Two concentrations of compounds per cell were tested.

Using internal and recording solutions results in a liquid junction potential of approximately 10 mV, which will be taken into account in the command voltage step.

Electrophysiological measurements: Membrane currents of cells were measured at distinct membrane potentials using the patch clamp technique by an automated patch clamp system. The holding potential is -70 mV. The hERG current (K ⁺ -selective outward current) was determined as the peak tail current at -30 mV after a 2 s depolarization to +70 mV (see 1, 4). The pulse cycling rate was 15 seconds.

data analysis

Leakage-corrected hERG current (K ⁺ selective outward current) was determined as the maximum tail current at -30 mV after 2 s depolarization to +70 mV measured between 2336.3 ms and 3083.6 ms. The median of the 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 SyncroPatch 384PE PatchControl384 software to automatically exclude wells from analysis if values were out of range. QC standards vary depending on the type of recording plate (chip). Typically, 4xChip (medium pore) was used to record from HEK293 cells transfected with hERG. QC criteria 4 to 6 were established prior to the first addition of compound; QC criteria 4 and 5 were also established at the end of each compound addition.

QC standards and acceptable ranges:

One. Board Check: -500pA to 500pA

2. Contact seal resistance: -100kOhm to 10MOhm

3. Junction potential offset: 0 to 100 mV

4. Reseal ≥ 100 MOhm

5. Rseries: 1 to 25 MOhm

6. hERG tail current ≥ 0.2 nA before compound addition

Each compound was replicated on the same plate in at least 5 wells. The median inhibition rate of at least 2 to 3 replicates per concentration will be reported. Results for Protocol 1 and Protocol 2 are summarized in Tables 14 and 15 below, respectively.

[Table 14]

[Table 15]

9) Efficacy study in disseminated OCI-AML3 model

Test agent and control group

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. The dose was adjusted daily depending on the individual's body weight. For each study, working stocks of compound A3 were prepared once per week and stored at 25°C.

animal

Female SCID beige mice (CB17.Cg-PrkdcscidLystbg-J/Crl/-) were used when they were approximately 6 to 8 weeks of age and weighed approximately 25 g. All animals were allowed to adapt and recover from transport-related stress for at least 7 days before being used in experiments. 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 before use and replaced weekly. Tissue culture and cell injection reagents are summarized in Table 16 below.

[Table 16]

Tumor models and cell culture methods

The human AML cell line OCI-AML3 was cultured in the indicated complete culture medium (MEM Alpha + 20% HI-FBS (heat-inactivated fetal bovine serum) + 2mM L-glutamine + 50ug/ml gentamicin) at 37°C in 5% CO ₂ It has been done. Cells were harvested in 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 ^5x10 cells via IV injection in a total volume of 0.2 mL using a 26-gauge needle.

study design

Compound A3 was administered orally (PO) daily.

Day 0 is the date of tumor cell transplantation and study initiation.

In the efficacy study, 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, 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 compound toxicity or tumor burden (i.e. hind limb paralysis, coma, etc.).

calculate

For survival assessment, results were plotted as percent survival against days after tumor implantation. Negative clinical signs and/or weight loss of more than 20% were used as surrogate endpoints for death. Median survival was determined using Kaplan-Meier survival analysis. The percent increased life span (ILS) was calculated as follows: ((Median survival days of treatment group - Median survival days of control group) / Median survival days of control group) × 100. Adverse clinical signs (e.g., ulcerated tumor, Animals that did not reach the surrogate endpoint due to weight loss, etc.) or death unrelated to treatment were censored for survival assessment. As defined by NCI criteria, ILS of 25% or greater is considered biologically significant. (Johnson JI et al. Br J Cancer. 2001. 84(10), 1424-1431]).

data analysis

Survival and body weight data were graphically displayed using Prism (version 7). Statistical significance for body weight was assessed as described above. Statistical significance was assessed for Kaplan-Meier survival plots comparing treatment groups with 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 the p value was 0.05 or less.

survival rate

Kaplan-Meier survival curves are shown in Figure 3. Mice bearing established OCI-AML3 tumors were orally administered 30, 50, or 100 mg/kg of Compound A3 in a 20% HP-β-CD formulation daily for a total of 28 days (n=9 to 10 mice/group). For the Compound A3 treatment group, median survival days were then reached at 75.5 days for 30 mg/kg, 50 mg/kg at 58.5 days, and 100 mg/kg at 75 days, compared to 38.5 days for the vehicle-treated control group. It was compared with the median survival value of . 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 control mice (p ≤0.001 ). This was biologically significant ILS according to the NCI standard threshold of ILS of 25% or more (Johnson JI et al. Br J Cancer. 2001. 84(10), 1424-1431).

10) Efficacy of menin-MLL inhibitors combined with venetoclax and optionally azacytidine in the OCI-AML3 (NPM1c) IV model in NSG mice

The disseminated OCI-AML3 model described above was also tested with venetoclax (formulated in polyethylene glycol [PEG]-400/Phosal50/dimethyl sulfoxide [DMSO]), and optionally azacytidine (formulated in 10% DMSO). ) in combination with a menin-MLL inhibitor (formulated as described above) with the following differences: (i) instead of female SCID beige, NSG (non-obesity diabetic [NOD]); scid gamma or NOD.Cg- Prkdc ^scid Il2rg ^tm1Wjl /SzJ) were utilized, (ii) instead of ⁵ ×10 cells, each mouse was injected with ¹ ×10 cells, and (iii) instead of 3 days, Treatment with compound(s) began 5 days after cell injection and (iv) compound (s) prepared to reach a volume of 8 ml/kg Compound A1, instead of a total volume of 0.2 mL (10 ml/kg) per dose. ) was given in the morning, and venetoclax or azacitidine was given in the afternoon at approximately 6 to 8 hour intervals. The treatment of each group with compound(s) in this efficacy study is summarized in Table 17.

[Table 17]

Kaplan-Meier survival curves are shown in Figure 4A. The effect on lifespan of 4 weeks of treatment of mice with established OCI-AML3 tumors is summarized in Table 18.

[Table 18]

As reflected in Table 18, treatment of mice with Compound A1 in combination with venetoclax or venetoclax + azacitine resulted in either vehicle control treated mice or azacitidine or venetoclax alone or venetoclax + aza There was a statistically significant increase in lifespan (ILS) of OCI-AML3 tumor-bearing mice compared to mice treated with the dual combination of cytidine. In particular, the triple combination (Compound A1 + venetoclax and azacitidine) or the double combination (venetoclax + azacitidine or Compound A1) has no antagonism and the efficacy appears to be entirely dependent on Compound A1.

11) Efficacy of menin-MLL inhibitors combined with venetoclax and optionally azacytidine in the MOLM-13 (KMT2A-r) IV model of NSG mice

A similar approach to the disseminated OCI-AML3 model described herein to investigate the efficacy of menin-MLL-inhibitors in combination with venetoclax and optionally azacytidine was followed by using MOLM-13 cells instead of OCI-AML3 cells. was adopted.

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 + 2mM L-glutamine + 50 μg/ml gentamicin) at 37°C in 5% CO _{2 .} did. Cells were harvested during logarithmic growth and resuspended in cold (4°C) serum-free RPMI 1640 medium. Each mouse was injected with 1 × 10 ⁵ MOLM-13 cells, and all other parameters were performed as described in the experiment (10).

The treatment of each group with compound(s) in this efficacy study is summarized in Table 19.

[Table 19]

Kaplan-Meier survival curves are shown in Figure 4B. The effect on lifespan of 4 weeks of treatment of mice with established MOLM-13 tumors is summarized in Table 20.

[Table 20]

As reflected in Table 20, treatment of mice with Compound A1 in combination with venetoclax or venetoclax + azacytidine resulted in lower MOLM compared to mice treated with venetoclax alone or a dual combination of venetoclax + azacitidine. -13 The lifespan (ILS) of tumor-bearing mice was statistically significantly increased.

(12) In vitro proliferation protocol

(A) Menin-MLL inhibitor in combination with venetoclax and optionally azacitidine

The effect of Compound A3 in combination with venetoclax or with venetoclax and azacytidine was determined in a proliferation assay using the MOLM-13 ( KMT2A-r ) cell line. MOLM-13 cells were seeded in 96 well plates at 500 cells/well and exposed to the indicated drugs alone or in combination at specific concentrations detailed in Tables 21a and 21b. In particular, for the combination of Compound A3 and venetoclax (without azacytidine), the concentration range of Compound A3 (7-point, 4-fold serial dilutions starting at an initial concentration of 1 μM) was compared to that of venetoclax (starting at an initial concentration of 1 μM). Starting with 6 points, 5-fold serial dilution) and combined. For the combination of Compound A3 with venetoclax and azacitidine, the concentration range of Compound A3 (6 points starting at an initial concentration of 1 μM, 4-fold serial dilution) was compared to that of azacytidine (7 points starting at an initial concentration of 25 μM). , 5-fold serial dilution) and venetoclax (7 points, 5-fold serial dilution, starting at an initial concentration of 1 μM) in a 25:1 ratio. Compound A3 and azacitidine (when added) were treated on day 0 and venetoclax was added on day 4. MOLM-13 cells were incubated four times with the indicated drug combination concentrations at 37°C, 5% CO ₂ for 8 days (Compound A3 and azacitidine) and 4 days (venetoclax).

Spheroid-like growth was monitored in real time by noninvasive live cell imaging using the 4× objective of the Incucyte ZOOM Live Cell Imaging System (Essen BioScience) and acquiring images at day 8. Confluence (%), 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 the % confluence of DMSO wells was used as the baseline response.

Potential combination effects were assessed on day 8 and synergistic effects were analyzed using the R-based Biochemically Intuitive Generalized Loewe (BIGL) model implemented as a best single agent (HSA) null model (Van der Borght 2017). Data from two independent experiments were pooled and analyzed.

result

The dual combination of Compound A3 and venetoclax significantly increased cell proliferation inhibition compared to Compound A3 or venetoclax monotherapy (see Figure 5a). As reflected in the contour plot (Figure 5A) and detailed in Table 21A below, a strong synergistic effect was observed across a range of concentrations of MOLM-13 cells, where the effect size is expressed as the difference between the observed and expected outcomes of the combination treatment; 95% confidence intervals are shown in parentheses.

The triple combination of Compound A3 with venetoclax and azacitidine significantly increased cell proliferation inhibition compared to Compound A3 monotherapy or the double 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 of MOLM-13 cells, where the effect size is expressed as the difference between the observed and expected results of the combination treatment; 95% confidence intervals are shown in parentheses.

[Table 21a]

[Table 21b]

(B) Menin-MLL inhibitor combined with decitabine and venetoclax

The effect of Compound A4 in combination with decitabine compared to the combination with decitabine and venetoclax was determined in proliferation assays using MOLM-13 ( KMT2A-r ) and OCI-AML3 (NPM1c) cell lines.

cell line

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 ribo- and deoxyribonucleosides) + 10-20% FBS. All cell lines were cultured at 37°C in a 5% CO ₂ atmosphere.

Cell Titer Glo Assay

AML cell lines MOLM-13 and OCI-AML3 ( ⁵ grown for 6 days. 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 to 4 independent experiments in technical triplicates. Compound A4 and decitabine were added on day 0 and venetoclax (if added) was added on day 4.

Synergy Calculation

The R-based Biochemically Intuitive Generalized Loewe (BIGL) model was implemented as the highest single agent (HSA) null model. Specifically, BIGL methodology was applied to calculate drug-drug interactions (see 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 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)]).

Synergy matrix results from BIGL analysis of cell line data using HSA as averaging model were calculated based on cell metabolic activity using the Cell Titer-Glo assay. Bootstrap confidence intervals are displayed. Effect sizes and confidence intervals are displayed. In particular, each data point is based on the p-value and sign of each maxR statistic, with the size of the point reflecting the degree of synergy or antagonism corresponding to the rating scale. If the interval includes 0, there is no significant average effect.

result

Pairwise matrix combinations of decitabine and Compound A4 in the absence or presence of venetoclax were used in MOLM-13 (KMT2A-AF9; FLT3-ITD) and OCI-AML3 (NPM1c AML) cells using a 6-day Cell Titer-Glo assay format. evaluated in In particular, the combination of decitabine and Compound A4, with or without venetoclax, resulted in MOLM-13 (KMT2A- showed synergistic cytotoxicity in AF9; FLT3-ITD) cells.

[Table 22a]

[Table 22b]

Antagonistic effects in OCI-AML3 (NPM1c AML) cells were reflected in the contour plots (Figures 7A and 7B) with and without venetoclax and at low dose decitabine (15 nM) as detailed in Tables 23A and 23B below. ) was observed.

[Table 23a]

[Table 23b]

SEQUENCE LISTING <110> Janssen Pharmaceutica N.V. Johnson & Johnson (China) Investment Ltd. <120> COMBINATION THERAPIES <130>P2022TC2023 <150> PCT/CN2021/093036 <151> 2021-05-11 <150> PCT/CN2021/100523 <151> 2021-06-17 <150> PCT/CN2022/086004 <151> 2022-04-11 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 616 <212> PRT <213> Artificial Sequence <400> 1 Met Gly Leu Lys Ala Ala Gln Lys Thr Leu Phe Pro Leu Arg Ser Ile 1 5 10 15 Asp Asp Val Val Arg Leu Phe Ala Ala Glu Leu Gly Arg Glu Glu Pro 20 25 30 Asp Leu Val Leu Leu Ser Leu Val Leu Gly Phe Val Glu His Phe Leu 35 40 45 Ala Val Asn Arg Val Ile Pro Thr Asn Val Pro Glu Leu Thr Phe Gln 50 55 60 Pro Ser Pro Ala Pro Asp Pro Pro Gly Gly Leu Thr Tyr Phe Pro Val 65 70 75 80 Ala Asp Leu Ser Ile Ile Ala Ala Leu Tyr Ala Arg Phe Thr Ala Gln 85 90 95 Ile Arg Gly Ala Val Asp Leu Ser Leu Tyr Pro Arg Glu Gly Gly Val 100 105 110 Ser Ser Arg Glu Leu Val Lys Lys Val Ser Asp Val Ile Trp Asn Ser 115 120 125 Leu Ser Arg Ser Tyr Phe Lys Asp Arg Ala His Ile Gln Ser Leu Phe 130 135 140 Ser Phe Ile Thr Gly Thr Lys Leu Asp Ser Ser Gly Val Ala Phe Ala 145 150 155 160 Val Val Gly Ala Cys Gln Ala Leu Gly Leu Arg Asp Val His Leu Ala 165 170 175 Leu Ser Glu Asp His Ala Trp Val Val Phe Gly Pro Asn Gly Glu Gln 180 185 190 Thr Ala Glu Val Thr Trp His Gly Lys Gly Asn Glu Asp Arg Arg Gly 195 200 205 Gln Thr Val Asn Ala Gly Val Ala Glu Arg Ser Trp Leu Tyr Leu Lys 210 215 220 Gly Ser Tyr Met Arg Cys Asp Arg Lys Met Glu Val Ala Phe Met Val 225 230 235 240 Cys Ala Ile Asn Pro Ser Ile Asp Leu His Thr Asp Ser Leu Glu Leu 245 250 255 Leu Gln Leu Gln Gln Lys Leu Leu Trp Leu Leu Tyr Asp Leu Gly His 260 265 270 Leu Glu Arg Tyr Pro Met Ala Leu Gly Asn Leu Ala Asp Leu Glu Glu 275 280 285 Leu Glu Pro Thr Pro Gly Arg Pro Asp Pro Leu Thr Leu Tyr His Lys 290 295 300 Gly Ile Ala Ser Ala Lys Thr Tyr Tyr Arg Asp Glu His Ile Tyr Pro 305 310 315 320 Tyr Met Tyr Leu Ala Gly Tyr His Cys Arg Asn Arg Asn Val Arg Glu 325 330 335 Ala Leu Gln Ala Trp Ala Asp Thr Ala Thr Val Ile Gln Asp Tyr Asn 340 345 350 Tyr Cys Arg Glu Asp Glu Glu Ile Tyr Lys Glu Phe Phe Glu Val Ala 355 360 365 Asn Asp Val Ile Pro Asn Leu Leu Lys Glu Ala Ala Ser Leu Leu Glu 370 375 380 Ala Gly Glu Glu Arg Pro Gly Glu Gln Ser Gln Gly Thr Gln Ser Gln 385 390 395 400 Gly Ser Ala Leu Gln Asp Pro Glu Cys Phe Ala His Leu Leu Arg Phe 405 410 415 Tyr Asp Gly Ile Cys Lys Trp Glu Glu Gly Ser Pro Thr Pro Val Leu 420 425 430 His Val Gly Trp Ala Thr Phe Leu Val Gln Ser Leu Gly Arg Phe Glu 435 440 445 Gly Gln Val Arg Gln Lys Val Arg Ile Val Ser Arg Glu Ala Glu Ala 450 455 460 Ala Glu Ala Glu Glu Pro Trp Gly Glu Glu Ala Arg Glu Gly Arg Arg 465 470 475 480 Arg Gly Pro Arg Arg Glu Ser Lys Pro Glu Glu Pro Pro Pro Pro Lys 485 490 495 Lys Pro Ala Leu Asp Lys Gly Leu Gly Thr Gly Gln Gly Ala Val Ser 500 505 510 Gly Pro Pro Arg Lys Pro Pro Gly Thr Val Ala Gly Thr Ala Arg Gly 515 520 525 Pro Glu Gly Gly Ser Thr Ala Gln Val Pro Ala Pro Ala Ala Ser Pro 530 535 540 Pro Pro Glu Gly Pro Val Leu Thr Phe Gln Ser Glu Lys Met Lys Gly 545 550 555 560 Met Lys Glu Leu Leu Val Ala Thr Lys Ile Asn Ser Ser Ala Ile Lys 565 570 575 Leu Gln Leu Thr Ala Gln Ser Gln Val Gln Met Lys Lys Gln Lys Val 580 585 590 Ser Thr Pro Ser Asp Tyr Thr Leu Ser Phe Leu Lys Arg Gln Arg Lys 595 600 605 Gly Leu His His His His His His 610 615

Claims (25)

A therapeutically effective amount of a menin-mixed lineage leukemia 1 (MLL) inhibitor of formula (I), or a tautomeric or stereoisomeric form thereof, or a pharmaceutically 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 antineoplastic agent
As a combination containing,
A combination wherein the menin-MLL inhibitor of formula (I) has the structure:
[Formula (I)]
,
In the above equation,
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; wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, -OC _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, X ² represents N;
R ⁴ 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 is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 alkyl; Here, each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is 1, 2, each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. may be substituted with 2 or 3 substituents;
R ^8a and R ^8b are each independently hydrogen; C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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. 2. The method of claim 1, wherein the menin-MLL inhibitor of Formula (I) is Compound A:

or a pharmaceutically acceptable salt or solvate thereof. 2. The method of claim 1, wherein the menin-MLL inhibitor of formula (I) is compound A4-a:

or a combination thereof, which is a solvate thereof. The method of any one of claims 1 to 3, wherein the BCL-2 inhibitor is obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax, gamgoic acid. A combination selected from (gamgogic acid), venetoclax, and pharmaceutically acceptable salts or solvates thereof. The combination of claim 4, wherein the BCL-2 inhibitor is venetoclax, or a pharmaceutically acceptable salt or solvate thereof. 6. The method 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 analog, a purine analog, a kinase inhibitor, a CD20 inhibitor, an isocitrate dehydrogenase inhibitor, A combination that is an immunomodulatory antineoplastic agent or a dihydroorotate dehydrogenase inhibitor. 7. The combination of claim 6, wherein at least one other antineoplastic agent is a hypomethylating agent. The combination according to claim 7, wherein the hypomethylating agent is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. The method of claim 1, wherein the menin-MLL inhibitor is Compound A or a pharmaceutically acceptable salt or solvate thereof, and the BCL-2 inhibitor is venetoclax, or a pharmaceutically acceptable salt or solvate thereof, and at least one The combination, wherein the other antineoplastic agent is a hypomethylating agent. The combination according to claim 9, wherein the hypomethylating agent is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. A pharmaceutical composition comprising a combination as claimed in any one of claims 1 to 10 and a pharmaceutically acceptable carrier. A combination as claimed in claims 1 to 10 or a pharmaceutical composition as claimed in claim 11 for use as a medicament. A combination as claimed in claims 1 to 10 or a pharmaceutical composition as claimed in claim 11 for use in the prevention or treatment, especially in the treatment, of hematopoietic disorders. 14. The combination or pharmaceutical composition according to claim 13, wherein the hematopoietic disorder is nucleophosmin 1 (NPM1) mutant leukemia or MLL rearranged leukemia. The combination or pharmaceutical composition according to claim 13, wherein the hematopoietic disorder is acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). A method of treating a subject diagnosed with a hematopoietic disorder, comprising:
A therapeutically effective amount of a menin-mixed lineage leukemia 1 (MLL) inhibitor of formula (I), or a tautomeric or stereoisomeric form thereof, or a pharmaceutically acceptable salt or solvate thereof;
A therapeutically effective amount of a BCL-2 inhibitor; and
Optionally, comprising administering a therapeutically effective amount of at least one other antineoplastic agent,
The menin-MLL inhibitor of formula (I) has the structure:
[Formula (I)]
,
In the above equation,
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; wherein the 5- or 6-membered monocyclic aromatic ring is optionally substituted with one or two 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, —OC _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, X ² represents N;
R ⁴ 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 is -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)-OC _1-4 alkyl-OC(=O)-C _1-4 represents alkyl; Here, each C _1-4 alkyl or C _1-6 alkyl moiety in the definition of R ³ is 1, 2, each independently selected from the group consisting of cyano, halo, -OH, and -OC _1-4 alkyl. may be substituted with 2 or 3 substituents;
R ^8a and R ^8b are each independently hydrogen;
C _1-6 alkyl; -C(=O)-C _1-4 alkyl; -C(=O)-OC _1-4 alkyl; -C(=O)-NR ^12a R ^12b ; and -OH, cyano, halo, -S(=O) ₂ -C _1-4 alkyl, -OC _1-4 alkyl, -C(=O)-NR ^10a R ^10b , and -NR ^10c -C(= O) is selected from the group consisting of C _1-6 alkyl substituted with 1, 2 or 3 substituents each independently selected from the group consisting of -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. 17. The method of claim 16, wherein the menin-MLL inhibitor of Formula (I) is Compound A:

or a pharmaceutically acceptable salt or solvate thereof. 12. The method of claim 11, wherein the menin-MLL inhibitor of formula (I) is compound A4-a:

or a solvate thereof. 19. The method of any one of claims 16 to 18, wherein the BCL-2 inhibitor is venetoclax, obatoclax, HA14-1, navitoclax, ABT-737, TW-37, AT101, sabutoclax. sulfate, gamgoic acid, and pharmaceutically acceptable salts or solvates thereof. The method of claim 19, wherein the BCL-2 inhibitor is venetoclax, or a pharmaceutically acceptable salt or solvate thereof. 21. 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, A method, which is an immunomodulatory antineoplastic agent or a dihydroorotate dehydrogenase inhibitor. 22. The method of claim 21, wherein at least one other antineoplastic agent is a hypomethylating agent. The method of claim 22, wherein the hypomethylating agent is azacytidine, or a pharmaceutically acceptable salt or solvate thereof. 24. The method of any one of claims 16-23, wherein the hematopoietic disorder is nucleophosmin 1 (NPM1) mutant leukemia or MLL rearranged leukemia. 24. The method of any one of claims 16-23, wherein the hematopoietic disorder is acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL).

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