Macrocyclic broad-spectrum antibiotics

Abstract

To provide a novel class of broad spectrum antibiotics for treating multidrug-resistant pathogens.SOLUTION: The invention provides lipopeptide macrocyclic compounds represented by the general formula in the figure.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 853,457, filed on 28 May 2019, which is incorporated herein by reference in its entirety for all purposes. [Background technology]

[0002] Antibiotic resistance is a serious and growing phenomenon in modern medicine and has emerged as a major public health concern in the 21st century. For example, certain antimicrobial compounds are described in International Patent Publication 2018 / 149419, the entirety of which is incorporated herein by reference. Some of these antimicrobial compounds are shown in Table 1. JPEG0007842812000001.jpg219167 JPEG0007842812000002.jpg213167 JPEG0007842812000003.jpg249167 JPEG0007842812000004.jpg250169

[0003] However, further novel classes of broad-spectrum antibiotics are still needed to treat certain multidrug-resistant pathogens. [Overview of the project]

[0004] New macrocyclic compounds for the treatment of microbial infections, such as bacterial infections, are described herein. In various embodiments, the present disclosure provides lipopeptide macrocyclic compounds for the treatment of bacterial infections. In various embodiments, the present disclosure provides classes and subclasses of chemical compounds structurally related to arylomycins for the treatment of bacterial infections. In various embodiments, the macrocyclic compound acts by inhibiting bacterial type I signal peptidase (SpsB), an essential protein of bacteria. In some embodiments, the signal peptidase is a Gram-negative signal peptidase. In some embodiments, the signal peptidase is LepB. The compounds of the present invention are useful for the treatment of Gram-negative bacterial infections and are particularly useful for the treatment of infections associated with non-fermenting bacteria.

[0005] In one aspect described herein, formula (I) JPEG0007842812000005.jpg69112 Formula (I); (wherein, R b , d , a , b , d , d , c , b , c , a , c is H, or -(C1-C6)alkyl optionally substituted with one, two or three R 1a ; Each R 1a is independently halogen, -CN, -OR a , -SR a , -NR c R d , -NO2, -C(=O)R b , -NR a C(=O)R b , -C(=O)OR a , -C(=O)NR c R d , -NR a C(=O)NR c R d , -S(=O)2R b , -S(=O)R b , -S(=O)2NR c R d , -S(=O)NR c R d , -NR a S(=O)2R b, -NR a S(=O)2NR c R d , -NR a Ure a , -NR a C(=O)NR a Ure a -O(C1-C6)alkylene-NR c R d , -NR a C(=NR c )R a -C(=NR a )NR c R d , -NR a C(=NR a )NR c R d , cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or two R on the same carbon 1a They come together to form an oxo; R 2 H, -NR c R d , or one, two or three R 2a It is a (C1-C6) alkyl group which may be substituted with; Each R 2a These are halogen, -CN, and -OR, independently. a , -SR a , -NR c R d -NO2, -C(=O)R b , -NR a C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -NR a C(=O)NR c R d -S(=O)2R b -S(=O)R b -S(=O)2NR c R d -S(=O)NR c R d , -NR a S(=O)2R b , -NR aS(=O)2NR c R d 、 -NR a OR a 、 -NR a C(=O)NR a OR a 、 -O(C1 - C6)alkylene - NR c R​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​, -NR a Ure a , -NR a C(=O)NR a Ure a -O(C1-C6)alkylene-NR c R d , -NR a C(=NR c )R a -C(=NR a )NR c R d , -NR a C(=NR a )NR c R d , cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or two R on the same carbon 3a They come together to form an oxo; R 4 is H or -(C1-C6) alkyl; X is (C1-C6)alkylene, (C2-C6)alkenylene, (C2-C6)alkynylene, (C3-C7)cycloalkylene, (C2-C7)heterocycloalkylene, arylene, or heteroarylene; alkylene, alkenylene, alkynylene, cycloalkylene, heteroacycloalkylene, arylene, and heteroarylene are one, two, or three R X It may also be replaced with; Each R X These are halogen, -CN, and -OR, independently. a , -NR c R d -NO2, -C(=O)R b , -C(=O)OR a -C(=O)NR c R d -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbon X They come together to form an oxo; Y is a bond, -O-, -S-, (C1-C6)alkylene, (C2-C6)alkenylene, (C2-C6)alkynylene, (C3-C7)cycloalkylene, (C2-C7)heterocycloalkylene, arylene, or heteroarylene; alkylene, alkenylene, alkynylene, cycloalkylene, heteroacycloalkylene, arylene, and heteroarylene have one, two, or three R Y It may also be replaced with; Each R Y These are halogen, -CN, and -OR, independently. a , -NR c R d -NO2, -C(=O)R b , -C(=O)OR a -C(=O)NR c R d -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbon Y They come together to form an oxo; Z stands for H, Halogen, -CN, -OR 10 , -SR 10 , -NR 12 R 13 -C(=O)R 11 , -C(=O)OR 2 -C(=O)NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 )aminoalkyl, -(C2-C 12 ) Alkenyl, -(C2-C 12) are alkynyl, -(C3-C9)cycloalkyl, -(C2-C7) heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heteroacycloalkyl, aryl, and heteroaryl are one, two, or three R Z It may also be replaced with; Each R Z These are halogen, -CN, and -OR, independently. 10 , -NR 12 R 13 -NO2, -C(=O)R 11 , -C(=O)OR 10 -C(=O)NR 12 R 13 -(C1-C6)alkyl, -(C1-C6)haloalkyl, (C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbon Z They come together to form an oxo; Each R 10 These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may have one, two, or three R 10a They may be substituted independently; Each R 10a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d-(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 10a They come together to form an oxo; Each R 11 These are independently -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are one, two, or three R 11a They may be substituted independently; Each R 11a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 11a They come together to form an oxo; Each R 12 and R 13 These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may have one, two, or three R 12a They may be substituted independently; Or, R 12 and R 13 Together with the nitrogen atom to which they are bonded, they form one, two, or three R atoms.12b It forms a heterocycloalkyl group which may be substituted with; Each R 12a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 12a They come together to form an oxo; Each R 12b These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 12b They come together to form an oxo; Each R a These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Each R bThese are independently -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Each R c and R d These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Or, R c and R d They, together with the nitrogen atom to which they are bonded, form a heterocycloalkyl which may be substituted with one, two, or three oxo, halogen, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl. The compound, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.

[0006] In some embodiments of the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or stereoisomers; the compound has the structure of formula (Ia). JPEG0007842812000006.jpg72112 formula (Ia)

[0007] In some embodiments of the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or stereoisomers; the compound has the structure of formula (Ib). JPEG0007842812000007.jpg69112 formula (Ib)

[0008] Pharmaceutical compositions comprising a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, and a pharmaceutically acceptable excipient are also disclosed herein.

[0009] Also disclosed herein is a method for treating bacterial infections in mammals, comprising administering to mammals an effective amount of the compounds disclosed herein, or pharmaceutically acceptable salts, solvates, or stereoisomers thereof, at a frequency and duration sufficient to produce a beneficial effect on the mammal.

[0010] Also disclosed herein is a method for treating lepB-borne infections in mammals, comprising administering to mammals an effective amount of the compounds disclosed herein, or pharmaceutically acceptable salts, solvates, or stereoisomers thereof, at a frequency and duration sufficient to produce a beneficial effect on the mammal. In some embodiments, bacterial infections include Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, and Salmonella typhimurium. Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, MorganellaMorganii), Proteus mirabilis, Proteus vulgaris, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia intermedia), Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobacter pylori Campylobacter pylori, Campylobacter fetusCampylobacter fetus, Campylobacter jejuni, Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella, Gardnerella vaginalis, Bacteroides fragilis Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides splanchnicus, Clostridium difficile, Mycobacterium tuberculosis, Mycobacterium avium Mycobacterium avium), Mycobacterium intracellulare, Mycobacterium lepraeleprae), Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcus hominis, or Staphylococcus saccharoticusThe infection is one in which *Saccharolyticus* is involved. In some embodiments of the treatment method, the bacterial infection is one in which *Acinetobacter baumannii*, *Klebsiella pneumoniae*, or *Pseudomonas erginosa* is involved. In some embodiments of the treatment method, the bacterial infection is one in which *Acinetobacter baumannii* is involved. In some embodiments of the treatment method, the bacterial infection is one in which *Gram-negative* bacteria are involved. In some embodiments of the treatment method, the method further comprises administering a second therapeutic agent. In some embodiments of the treatment method, the second therapeutic agent is not an SpsB or LepB inhibitor. In some embodiments of the treatment method, the second therapeutic agent is an aminoglycoside antibiotic, a fluoroquinolone antibiotic, a β-lactam antibiotic, a macrolide antibiotic, a glycopeptide antibiotic, rifampicin, chloramphenicol, fluoramphenicol, colistin, mupirocin, bacitracin, daptomycin, or linezolid. The method according to claim 48, wherein the second therapeutic agent is a β-lactam antibiotic. The method according to claim 50, wherein the β-lactam antibiotic is selected from penicillin, monobactam, cephalosporin, cephamycin and carbapenem. In some embodiments of the treatment method, the β-lactam antibiotic is azurocillin, amoxicillin, ampicillin, doripenem, meropenem, biapenem, cephamandol, imipenem, mezlocillin, cefmetazole, cefprodil, piperacillin / tazobactam, carbenicillin, cefaclor, cephalothin, ertapenem, cefazolin, cefepime, cefonisid, The following are selected from cefoxitine, ceftazidime, oxacillin, cefdinir, cefixime, cefotaxime, cefotetan, cefpodoxime, ceftizoxime, ceftriaxone, faropenem, mecillinam, methicillin, moxalactam, ticarcillin, tomopenem, ceftobiprole, cephthaloline, flomoxef, cefpirome (Cefiprome), and cefozopran. In some embodiments of the treatment method, the method further comprises administering a β-lactamase inhibitor.

[0011] Built-in by reference All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as each individual publication, patent, or patent application is incorporated herein by reference specifically and individually. Detailed Description of the Invention

[0012] definition When used herein and in the appended claims, the singular forms “a,” “and,” and “the” include multiple references unless the context explicitly indicates otherwise. For example, a reference to “an agent” includes multiple such agents, and a reference to “the cell” includes one or more cells (or more cells) and their equivalents, etc., known to those skilled in the art. Where a range is used herein for physical properties such as molecular weight, or chemical properties such as chemical formula, it is intended to include all combinations and partial combinations of the range and particular embodiments. When referring to a number or numerical range, the term “about” means that the number or numerical range referred to is an approximation within experimental variability (or statistical experimental error), and therefore, in some examples, the number or numerical range varies between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise,” “comprises,” “having,” or “including”) is not intended to exclude the described features from “consisting of” or “consisting essentially of” in other specific embodiments, such as the composition, composition, method, or process of any substance described herein.

[0013] As used herein and in the appended claims, unless otherwise specified, the following terms have the meanings set forth below.

[0014] "Alkyl" refers to a substituted or substituted linear or substituted branched saturated hydrocarbon monoradical having 1 to about 10 carbon atoms, or 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, tert-amyl, and hexyl, as well as longer alkyl groups such as heptyl and octyl. Wherever it appears herein, numerical ranges such as "C1-C6 alkyl" mean that the alkyl group consists of one, two, three, four, five, or six carbon atoms; however, this definition also includes instances where the term "alkyl" does not specify a numerical range. In some embodiments, alkyl is C1-C 20 Alkyl, C1-C 10 The alkyl group is alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or C1 alkyl. Unless otherwise specified herein, alkyl groups may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, alkyl may be substituted with oxo, halogen, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, alkyl may be substituted with oxo, halogen, -CN, -CF3, -OH, or -OMe. In some embodiments, alkyl may be substituted with halogen.

[0015] "Alkenyl" refers to a substituted or substituted linear or substituted branched hydrocarbon monoradical having one or more carbon-carbon double bonds and having 2 to about 10 carbon atoms, more preferably 2 to about 6 carbon atoms. The group may be in either cis or trans configuration around the double bond and should be understood to include both isomers. Examples include, but are not limited to, ethenyl (-CH=CH2), 1-propenyl (-CH2CH=CH2), isopropenyl [-C(CH3)=CH2], butenyl, 1,3-butadienyl, etc. Whenever it appears herein, numerical ranges such as "C2-C6 alkenyl" mean that the alkenyl group may consist of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms, but this definition also includes appearances of the term "alkenyl" where no numerical range is specified. In some embodiments, the alkenyl is C2-C 20 Alkenyl, C2-C 10 The alkenyl group is an alkenyl, a C2-C9 alkenyl, a C2-C8 alkenyl, a C2-C7 alkenyl, a C2-C6 alkenyl, a C2-C5 alkenyl, a C2-C4 alkenyl, a C2-C3 alkenyl, or a C2 alkenyl. Unless otherwise specified herein, the alkenyl group may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the alkenyl may be substituted with oxo, halogen, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the alkenyl may be substituted with oxo, halogen, -CN, -CF3, -OH, or -OMe. In some embodiments, the alkenyl may be substituted with halogen.

[0016] "Alkynyl" refers to a substituted or substituted linear or substituted branched hydrocarbon monoradical having one or more carbon-carbon triple bonds and containing 2 to about 10 carbon atoms, more preferably 2 to about 6 carbon atoms. Examples include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1,3-butadiinyl. Wherever it appears herein, numerical ranges such as "C2-C6 alkynyl" mean that the alkynyl group may consist of 2, 3, 4, 5, or 6 carbon atoms; however, this definition also includes instances where the term "alkynyl" does not specify a numerical range. In some embodiments, alkynyl is C2-C 20 Alkinyl, C2-C 10 These are alkynyl, C2-C9 alkynyl, C2-C8 alkynyl, C2-C7 alkynyl, C2-C6 alkynyl, C2-C5 alkynyl, C2-C4 alkynyl, C2-C3 alkynyl, or C2 alkynyl. Unless otherwise specified herein, the alkynyl group may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the alkynyl may be substituted with oxo, halogen, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the alkynyl may be substituted with oxo, halogen, -CN, -CF3, -OH, or -OMe. In some embodiments, the alkynyl may be substituted with halogen.

[0017] "Alkylene" refers to a straight-chain or branched-chain divalent hydrocarbon chain. Unless otherwise specified herein, the alkylene group may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the alkylene may be substituted with oxo, halogen, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the alkylene may be substituted with oxo, halogen, -CN, -CF3, -OH, or -OMe. In some embodiments, the alkylene may be substituted with halogen.

[0018] "alkoxy" is the formula -OR a It refers to the base of, and in the formula, R a is an alkyl group as defined. Unless otherwise specified herein, the alkoxy group may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the alkoxy may be substituted with oxo, halogen, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the alkoxy may be substituted with oxo, halogen, -CN, -CF3, -OH, or -OMe. In some embodiments, the alkoxy may be substituted with halogen.

[0019] "Aryl" refers to a group derived from a hydrocarbon ring system containing hydrogen, 6 to 30 carbon atoms, and at least one aromatic ring. Aryl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, and may include condensed ring systems (where the aryl is bonded via aromatic ring atoms when condensed with a cycloalkyl or heterocycloalkyl ring) or bridged ring systems. In some embodiments, the aryl is a 6- to 10-membered aryl. In some embodiments, the aryl is a 6-membered aryl. Aryl groups include, but are not limited to, aryl groups derived from the hydrocarbon ring systems of anthrylene, naphthylene, phenanthrylene, anthracene, azulene, benzene, chrysene, fluorantene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. In some embodiments, the aryl is phenyl. Unless otherwise specified herein, aryl may be substituted with, for example, halogens, aminos, nitriles, nitros, hydroxyls, alkyls, alkenyls, alkynyls, haloalkyls, alkoxys, aryls, cycloalkyls, heterocycloalkyls, heteroaryls, etc., as described below. In some embodiments, aryl may be substituted with halogens, methyls, ethyls, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, aryl may be substituted with halogens, methyls, ethyls, -CN, -CF3, -OH, or -OMe. In some embodiments, aryl may be substituted with halogens or methyls. In some embodiments, aryl may be substituted with halogens.

[0020] "Cycloalkyl" refers to a stable, partially or fully saturated, monocyclic or polycyclic carbocyclic ring, which may include condensed ring systems (where the cycloalkyl is condensed with an aryl or heteroaryl ring, the cycloalkyl is bonded via non-aromatic ring atoms) or bridged ring systems. Typical cycloalkyls have 3 to 15 carbon atoms (C3-C3). 15 Cycloalkyl), 3 to 10 carbon atoms (C3-C10 This includes, but is not limited to, cycloalkyls having 3 to 8 carbon atoms (C3-C8 cycloalkyl), 3 to 6 carbon atoms (C3-C6 cycloalkyl), 3 to 5 carbon atoms (C3-C5 cycloalkyl), or 3 to 4 carbon atoms (C3-C4 cycloalkyl). In some embodiments, the cycloalkyl is a 3- to 6-membered cycloalkyl. In some embodiments, the cycloalkyl is a 5- to 6-membered cycloalkyl. Examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyl or carbocyclic compounds include adamantyl, norbornyl, dekalinyl, bicyclo[3.3.0]octane, bicyclo[4.3.0]nonane, cis-decalin, trans-decalin, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane and bicyclo[3.3.2]decane, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Examples of partially saturated cycloalkyl compounds include cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. Unless otherwise specified herein, cycloalkyl compounds may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the cycloalkyl group may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the cycloalkyl group may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, or -OMe. In some embodiments, the cycloalkyl group may be substituted with halogen or methyl. In some embodiments, the cycloalkyl group may be substituted with halogen.

[0021] "Halo" or "halogen" refers to bromo, chloro, fluoro, or iodine. In some embodiments, the halogen is fluoro or chloro. In some embodiments, the halogen is fluoro.

[0022] "Haloalkyl" refers to an alkyl group as defined above, which is substituted by one or more halo groups as defined above, such as trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc.

[0023] "Aminoalkyl" refers to the alkyl radical as defined above, which is substituted with one or more -NH2 groups, such as -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH(NH2)CH3, -CH2CH(NH2)CH3, -CH(NH2)CH2CH3, etc.

[0024] "Hydroxyalkyl" refers to the alkyl radical defined above, which is substituted with one or more -OH groups, such as -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH(OH)CH3, -CH2CH(OH)CH3, -CH(OH)CH2CH3, etc.

[0025] "Heterocycloalkyl" refers to a stable 3- to 24-membered, partially or fully saturated ring group comprising 2 to 23 carbon atoms and 1 to 8 heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorus, and sulfur. Unless otherwise specifically indicated herein, heterocycloalkyl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and may include condensed ring systems (where condensed with an aryl ring or heteroaryl ring, the heterocycloalkyl is linked via a non-aromatic ring atom) or bridged ring systems, the nitrogen, carbon, or sulfur atoms in the heterocycloalkyl can optionally be oxidized, and the nitrogen atom can optionally be quaternized. In some embodiments, the heterocycloalkyl is a 3- to 6-membered heterocycloalkyl. In some embodiments, the heterocycloalkyl is a 5- to 6-membered heterocycloalkyl. In some embodiments, the heterocycloalkyl is a 6-membered heterocycloalkyl. In some embodiments, the heterocycloalkyl is a 5-membered heterocycloalkyl. In some embodiments, the heterocycloalkyl is a (C2-C7) heterocycloalkyl. Examples of such heterocycloalkyls include azilidinyl, azetidinyl, dioxolanil, thienyl[1,3]dithianil, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperadinyl, 4-piperidonyl, pyrrolidinyl, Examples include, but are not limited to, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianil, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, 1,3-dihydroisobenzofuran-1-yl, 3-oxo-1,3-dihydroisobenzofuran-1-yl, methyl-2-oxo-1,3-dioxosol-4-yl, and 2-oxo-1,3-dioxosol-4-yl. The term heterocycloalkyl also includes all cyclic forms of carbohydrates, including but not limited to monosaccharides, disaccharides, and oligosaccharides.Unless otherwise specified, heterocycloalkyls have 2 to 10 carbon atoms in the ring. When referring to the number of carbon atoms in a heterocycloalkyl, it should be understood that the number of carbon atoms in a heterocycloalkyl is not the same as the total number of atoms constituting the heterocycloalkyl (including heteroatoms) (i.e., the skeletal atoms of the heterocycloalkyl ring). Unless otherwise specifically indicated herein, heterocycloalkyls may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, heterocycloalkyls may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, heterocycloalkyls may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, or -OMe. In some embodiments, heterocycloalkyls may be substituted with halogen or methyl. In some embodiments, the heterocycloalkyl group may be substituted with a halogen.

[0026] "Heteroalkyl" refers to an alkyl group in which one or more of the alkyl backbone atoms are atoms other than carbon, such as oxygen, nitrogen (e.g., -NH-, -N(alkyl)-), sulfur, or a combination thereof. In a heteroalkyl group, the heteroalkyl is bonded to the rest of the molecule at the carbon atom of the heteroalkyl. In one embodiment, the heteroalkyl is a C1-C6 heteroalkyl having one, two, or three heteroatoms selected from oxygen, nitrogen (e.g., -NH-, -N(alkyl)-), or sulfur. In one embodiment, the heteroalkyl is a C1-C6 heteroalkyl having one or two heteroatoms selected from oxygen, nitrogen (e.g., -NH-, -N(alkyl)-), or sulfur. Unless otherwise specifically indicated herein, heteroalkyl may be substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc., as described below. In some embodiments, the heteroalkyl group may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the heteroalkyl group may be substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, or -OMe. In some embodiments, the heteroalkyl group may be substituted with halogen.

[0027] A "heteroaryl" refers to a 5- to 14-membered ring system comprising a hydrogen atom, 5 to 13 carbon atoms, 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorus, and sulfur, and at least one aromatic ring. A heteroaryl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and may include a condensed ring system (where the heteroaryl is condensed with a cycloalkyl ring or heterocycloalkyl ring, the heteroaryl is bonded via an aromatic ring atom) or a bridged ring system. The nitrogen, carbon, or sulfur atoms in the heteroaryl group may be optionally oxidized, and the nitrogen atom may be optionally quaternized. In some embodiments, the heteroaryl is a 5- to 10-membered heteroaryl. In some embodiments, the heteroaryl is a 5- to 6-membered heteroaryl.Examples include azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzoindolyl, benzodioxolyl, benzofuranil, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanil, benzonaphthofuranil, benzoxazolyl, benzodioxolyl, benzodioxynil, benzopyranil, benzopyranonil, benzofuranil, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, sinnolinil, dibenzofuranil, dibenzothiophenyl, furanil, furanonil, isothiazolyl, imidazolyl, indazolyl, indazolyl, isoindolyl, in Examples include, but are not limited to, dolinyl, isoindolinyl, isoquinolyl, indolidinyl, isoxazolyl, naphthilidinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxyranil, 1-oxidepyridinyl, 1-oxidepyrimidinyl, 1-oxidepyradinyl, 1-oxidepyridazinyl, 1-phenyl-1H-pyrrolyl, phenadinyl, phenothiazinyl, phenoxadinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridadinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless otherwise specified herein, heteroaryls may be substituted with, for example, halogens, aminos, nitriles, nitros, hydroxyls, alkyls, alkenyls, alkynyls, haloalkyls, alkoxys, aryls, cycloalkyls, heterocycloalkyls, heteroaryls, etc., as described below. In some embodiments, heteroaryls may be substituted with halogens, methyls, ethyls, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, heteroaryls may be substituted with halogens, methyls, ethyls, -CN, -CF3, -OH, or -OMe.In some embodiments, the heteroaryl may be substituted with a halogen or methyl group.

[0028] The term "Oxo" means =O.

[0029] As used herein, the terms “treat,” “prevent,” “improve,” and “inhibit,” and words derived therefrom, do not necessarily imply 100% or complete treatment, prevention, improvement, or inhibition. Rather, there are various degrees of treatment, prevention, improvement, and inhibition that a person skilled in the art would recognize as having a potential benefit or therapeutic effect. In this regard, the disclosed methods can provide any amount of treatment, prevention, improvement, or inhibition of any level of disorder in mammals. For example, a disorder, including its symptoms or manifestations, may be reduced by, for example, about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10%. Furthermore, the treatment, prevention, improvement, or inhibition provided by the methods disclosed herein may include treatment, prevention, improvement, or inhibition of one or more symptoms or manifestations of a disorder, such as cancer or an inflammatory disease. Also herein, “treatment,” “prevention,” “improvement,” or “inhibition” encompasses delaying the onset of a disorder, or its symptoms or manifestations.

[0030] As used herein, the terms “effective dose” or “therapeutic effective dose” refer to a sufficient amount of the compound disclosed herein administered to alleviate, to some extent, one or more of the symptoms of a disease or condition being treated, such as cancer or an inflammatory disease. In some embodiments, the result is a reduction and / or mitigation of the signs, symptoms or causes of the disease, or any other desired change in the biological system. For example, an “effective dose” for therapeutic use is the amount of a composition containing the compound disclosed herein required to produce a clinically significant reduction in the symptoms of the disease. In some embodiments, the appropriate “effective” dose in any individual case is determined using techniques such as dose-escalation studies.

[0031] As used herein, “individual” (as in the subject of treatment) means both mammals and non-mammals. Examples of mammals include humans, non-human primates such as apes and monkeys, and non-primates such as dogs, cats, cattle, horses, sheep and goats. Examples of non-mammals include fish and birds.

[0032] The terms “disease,” “disorder,” or “condition” are used interchangeably to refer to a disease or condition in which a bacterial SPase plays a role in the biochemical mechanism involved in the disease or disorder, such that a therapeutically beneficial effect can be achieved by acting on an enzyme. “Acting on” an SPase may include binding to and / or inhibition of the biological activity of the SPase.

[0033] As used herein, the term “prodrug” means a compound having one or more moieties that can be metabolized in vivo. For example, a prodrug is metabolized in vivo to an active drug by an esterase or other mechanism. Examples of prodrugs and their uses are well known in the art (see, for example, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19). Prodrugs can be prepared in situ during the final isolation and purification of a compound, or by reacting the purified compound individually with a suitable esterifying agent in its free acid form or hydroxyl group. The hydroxyl group can be converted to an ester via treatment with a carboxylic acid. Examples of prodrug moieties include substituted and unsubstituted, branched or unbranched lower alkyl ester moieties (e.g., propionic acid esters), lower alkenyl esters, dilower alkylamino lower alkyl esters (e.g., dimethylaminoethyl esters), acylamino lower alkyl esters (e.g., acetyloxymethyl esters), acyloxy lower alkyl esters (e.g., pivaloyloxymethyl esters), aryl esters (phenyl esters), aryl lower alkyl esters (e.g., benzyl esters), substituted aryl and aryl lower alkyl esters (e.g., with methyl, halo, or methoxy substituents), amides, lower alkylamides, dilower alkylamides, and hydroxyamides.

[0034] As used herein, the term “substantially” means completely or nearly completely. For example, a composition “substantially” free of an ingredient is either completely free of the ingredient or contains such a trace amount that any relevant functional properties of the composition are not affected by its presence, or a compound is “substantially pure” if it contains only a negligible amount of impurities.

[0035] compound In one embodiment described herein, formula (I): JPEG0007842812000008.jpg69112 formula (I); (In the formula, R 1 is H, or one, two or three R 1a It is a (C1-C6) alkyl group which may be substituted with; Each R 1a These are halogen, -CN, and -OR, independently. a , -SR a , -NR c R d -NO2, -C(=O)R b , -NR a C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -NR a C(=O)NR c R d -S(=O)2R b -S(=O)R b -S(=O)2NR c R d -S(=O)NR c R d , -NR a S(=O)2R b , -NR a S(=O)2NR c R d , -NR a Ure a , -NR a C(=O)NR a Ure a -O(C1-C6)alkylene-NR c R d , -NR a C(=NR c )R a -C(=NR a )NR c R d , -NR a C(=NR a )NR c R d , cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or two R on the same carbon 1aThey come together to form an oxo; R 2 H, -NR c R d , or one, two or three R 2a It is a (C1-C6) alkyl group which may be substituted with; Each R 2a These are halogen, -CN, and -OR, independently. a , -SR a , -NR c R d -NO2, -C(=O)R b , -NR a C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -NR a C(=O)NR c R d -S(=O)2R b -S(=O)R b -S(=O)2NR c R d -S(=O)NR c R d , -NR a S(=O)2R b , -NR a S(=O)2NR c R d , -NR a Ure a , -NR a C(=O)NR a Ure a -O(C1-C6)alkylene-NR c R d , -NR a C(=NR c )R a -C(=NR a )NR c R d , -NR a C(=NR a )NR c R d , cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or two R on the same carbon 2a They come together to form an oxo; R 3 is H, -(C3-C6) cycloalkyl, or one, two or three R 3a It is a (C1-C6) alkyl group which may be substituted with; Each R 3a These are halogen, -CN, and -OR, independently. a , -SR a , -NR c R d -NO2, -C(=O)R b , -NR a C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -NR a C(=O)NR c R d -S(=O)2R b -S(=O)R b -S(=O)2NR c R d -S(=O)NR c R d , -NR a S(=O)2R b , -NR a S(=O)2NR c R d , -NR a Ure a , -NR a C(=O)NR a Ure a -O(C1-C6)alkylene-NR c R d , -NR a C(=NR c )R a -C(=NR a )NR c R d , -NR a C(=NR a )NR c R d , cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or two R on the same carbon 3a They come together to form an oxo; R 4is H or -(C1-C6) alkyl; X is (C1-C6)alkylene, (C2-C6)alkenylene, (C2-C6)alkynylene, (C3-C7)cycloalkylene, (C2-C7)heterocycloalkylene, arylene, or heteroarylene; alkylene, alkenylene, alkynylene, cycloalkylene, heteroacycloalkylene, arylene, and heteroarylene are one, two, or three R X It may also be replaced with; Each R X These are halogen, -CN, and -OR, independently. a , -NR c R d -NO2, -C(=O)R b , -C(=O)OR a -C(=O)NR c R d -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbon X They come together to form an oxo; Y is a bond, -O-, -S-, (C1-C6)alkylene, (C2-C6)alkenylene, (C2-C6)alkynylene, (C3-C7)cycloalkylene, (C2-C7)heterocycloalkylene, arylene, or heteroarylene; alkylene, alkenylene, alkynylene, cycloalkylene, heteroacycloalkylene, arylene, and heteroarylene have one, two, or three R Y It may also be replaced with; Each R Y These are halogen, -CN, and -OR, independently. a , -NR c R d -NO2, -C(=O)R b , -C(=O)OR a -C(=O)NR c R d-(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbon Y They come together to form an oxo; Z stands for H, Halogen, -CN, -OR 10 , -SR 10 , -NR 12 R 13 -C(=O)R 11 , -C(=O)OR 2 -C(=O)NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 )aminoalkyl, -(C2-C 12 ) Alkenyl, -(C2-C 12 ) are alkynyl, -(C3-C9)cycloalkyl, -(C2-C7) heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heteroacycloalkyl, aryl, and heteroaryl are one, two, or three R Z It may also be replaced with; Each R Z These are halogen, -CN, and -OR, independently. 10 , -NR 12 R 13 -NO2, -C(=O)R 11 , -C(=O)OR 10 -C(=O)NR 12 R 13 -(C1-C6)alkyl, -(C1-C6)haloalkyl, (C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl or -(C2-C7)heterocycloalkyl; or two R on the same carbonZ They come together to form an oxo; Each R 10 These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may have one, two, or three R 10a They may be substituted independently; Each R 10a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 10a They come together to form an oxo; Each R 11 These are independently -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are one, two, or three R 11a They may be substituted independently; Each R 11a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NRc R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 11a They come together to form an oxo; Each R 12 and R 13 These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may have one, two, or three R 12a They may be substituted independently; Or, R 12 and R 13 Together with the nitrogen atom to which they are bonded, they form one, two, or three R atoms. 12b It forms a heterocycloalkyl group which may be substituted with; Each R 12a These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon 12a They come together to form an oxo; Each R 12b These are halogen, -CN, and -OR, independently. a , -NR c R d -C(=O)R b , -C(=O)OR a -C(=O)NR c R d , -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon12b They come together to form an oxo; Each R a These are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Each R b These are independently -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Each R c and R dThese are independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, -(C1-C6)heteroalkyl, -(C1-C6)hydroxyalkyl, -(C1-C6)aminoalkyl, -(C2-C6)alkenyl, -(C2-C6)alkynyl, -(C3-C7)cycloalkyl, (C2-C7)heterocycloalkyl, aryl, or heteroaryl; alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be independently substituted with one, two, or three halogens, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl; Or, R c and R d They, together with the nitrogen atom to which they are bonded, form a heterocycloalkyl which may be substituted with one, two, or three oxo, halogen, -CN, -OH, -OMe, -NH2, -C(=O)Me, -C(=O)OH, -C(=O)OMe, -C(=O)NH2, or -(C1-C6)alkyl. The compound, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.

[0036] In some embodiments of the compound of formula (I), R 2 is one, two, or three R 2a It is H or -(C1-C6)alkyl which may be substituted with. In some embodiments of the compound of formula (I), R 2 is H or one R 2a It is a -(C1-C6) alkyl which may be substituted with R. In some embodiments of the compound of formula (I), 2 is H or -(C1-C6) alkyl. In some embodiments of the compound of formula (I), R 2 H is H.

[0037] In some embodiments of the compound of formula (I), each R 2a These are, independently, halogen, -CN, and -OR. a , -NR c Rd , -NR c C(=O)R b , -NR c C(=O)NR c R d , -NR c S(=O)2R b , or -NR c S(=O)2NR c R d In some embodiments of the compound of formula (I), each R 2a -NR is independent. c R d or -NR c S(=O)2NR c R d That is the case.

[0038] In some embodiments of the compound of formula (I), R 3 is one, two, or three R 3a It is H or -(C1-C6)alkyl which may be substituted with. In some embodiments of the compound of formula (I), R 3 is one R 3a It is H or -(C1-C6)alkyl which may be substituted with. In some embodiments of the compound of formula (I), R 3 is H or -(C1-C6) alkyl. In some embodiments of the compound of formula (I), R 3 is H. In some embodiments of the compound of formula (I), R 3 R is a -(C1-C6) alkyl group. In some embodiments of the compound of formula (I), R 3 It is methyl.

[0039] In some embodiments of the compound of formula (I), each R 3a These are, independently, halogen, -CN, and -OR. a , -NR c R d , -NR c C(=O)R b , -NR c C(=O)NR c R d , -NR c S(=O)2R b , or -NRc S(=O)2NR c R d In some embodiments of the compound of formula (I), each R 3a -NR is independent. c R d or -NR c S(=O)2NR c R d That is the case.

[0040] In some embodiments of the compound of formula (I), R 4 is H. In some embodiments of the compound of formula (I), R 4 is a -(C1-C6) alkyl group. In some embodiments of the compound of formula (I), R 4 It is methyl.

[0041] In some embodiments of the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or stereoisomers; the compound has the structure of formula (Ia). JPEG0007842812000009.jpg72112 formula (Ia)

[0042] In some embodiments of the compound of formula (I), or its pharmaceutically acceptable salts, solvates, or stereoisomers; the compound has the structure of formula (Ib). JPEG0007842812000010.jpg69112 formula (Ib)

[0043] In some embodiments of compounds of formula (I), (Ia), or (Ib), R 1 is one, two, or three R 1a It is a -(C1-C6) alkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 is one or two R 1a It is a -(C1-C6) alkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 is one R 1aIt is a -(C1-C6) alkyl substituted with R. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 R is -CH2CH2NH2, -CH2CH2NHSO2NH2, or -CH2NHSO2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 is -CH2CH2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 is -CH2CH2NHSO2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 1 It is -CH2NHSO2NH2.

[0044] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R 1a These are, independently, halogen, -CN, and -OR. a , -NR c R d , -NR c C(=O)R b , -NR c C(=O)NR c R d , -NR c S(=O)2R b , or -NR c S(=O)2NR c R d In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R 1a -NR is independent. c R d or -NR c S(=O)2NR c R d In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R 1a -NR is independent c R d In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R 1a -NR is independent c S(=O)2NR c R d That is the case.

[0045] In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one, two, or three R X It is a (C2-C7) heterocycloalkylene or heteroarylene which may be substituted with R. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is a (C2-C7) heterocycloalkylene or heteroarylene, and one or two R X It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is a (C2-C7) heterocycloalkylene or heteroarylene, and one R X It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one, two, or three R X It is a (C2-C7) heterocycloalkylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one or two R X It is a (C2-C7) heterocycloalkylene which may be substituted with . In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one R X It is a (C2-C7) heterocycloalkylene which may be substituted with R. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one, two, or three R X It is a heteroarylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one or two R X It is a heteroarylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one R X It is a heteroarylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one or two R X It is a pyrimidinylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), X is one, two, or three R X It is a pyridinylene which may be substituted with

[0046] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X These are, independently, halogen, -OR a , -NR c R d -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon X These combine to form an oxo. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X -NR is independent. c R d or -(C1-C6) alkyl; or two R on the same carbon X These combine to form an oxo. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X -NR is independent. c R d or -(C1-C6)alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X R is independently Cl, -CN, methyl, ethyl, -CF2H, -CF3, -CH2NH2, cyclopropyl, or 3-aminoazetidine-1-yl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X R is independently -Cl, -CN, methyl, or -CH2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X R is independently -Cl, methyl, or -CH2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X R is independently -methyl or -CH2NH2. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X is 3-aminoazetidine-1-yl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R X It is independently methyl.

[0047] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is a -(C2-C7) heterocycloalkylene or arylene; each contains one, two, or three R Y It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is -(C2-C7) heterocycloalkylene or arylene. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is -(C2-C7) heterocycloalkylene or arylene; each of which has one or two R Y It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is a -(C2-C7) heterocycloalkylene or arylene; each is one R Y It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is one, two, or three R Y It is an arylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is one or two R Y It is an arylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is one R Y It is an arylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is an arylene. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is one, two, or three R Y It is a phenylene which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Y is a bond.

[0048] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Y These are, independently, halogen, -OR a , -NR c R d -(C1-C6)alkyl or -(C1-C6)haloalkyl; or two R on the same carbon YThese combine to form an oxo. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Y These are, independently, halogen, -OR a , -NR c R d It is -(C1-C6)alkyl or -(C1-C6)haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Y R is independently a halogen, a -(C1-C6)alkyl, or a -(C1-C6)haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Y R is independently F or -OH. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Y It is F.

[0049] In some embodiments of formula (I), (Ia), or (Ib), Z is H, halogen, -CN, -OR 10 , -SR 10 , -NR 12 R 13 -C(=O)R 11 , -C(=O)OR 2 -C(=O)NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 )aminoalkyl, -(C2-C 12 ) Alkenyl, -(C2-C 12 ) Alkynyl, -(C3-C7)cycloalkyl, -(C2-C7)heterocycloalkyl, aryl, or heteroaryl, and alkyl, alkenyl, alkynyl, cycloalkyl, heteroacycloalkyl, aryl, and heteroaryl have one, two, or three R Z It may be replaced with .

[0050] In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C9)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are 1, 2, or 3 R Z It may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C9)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are one or two R Z It may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C9)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are one R ZIt may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C9)cycloalkyl, or -(C2-C7)heterocycloalkyl.

[0051] In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C7)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are 1, 2, or 3 R Z It may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C7)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are one or two R Z It may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C7)cycloalkyl, or -(C2-C7)heterocycloalkyl; alkyl, cycloalkyl and heteroacycloalkyl are one R Z It may be replaced by: In some embodiments of formula (I), (Ia), or (Ib), Z is a halogen, -OR 10 , -NR 12 R 13 ,-(C1-C 12 )alkyl, -(C1-C 12 ) Heteroalkyl, -(C1-C 12 ) Haloalkyl, -(C1-C 12 ) Hydroxyalkyl, -(C1-C 12 ) are aminoalkyl, -(C3-C7)cycloalkyl, or -(C2-C7)heterocycloalkyl.

[0052] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C9)cycloalkyl; alkyl and cycloalkyl are one, two, or three R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C9)cycloalkyl; alkyl and cycloalkyl are one or two R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12)alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C9)cycloalkyl; alkyl and cycloalkyl are one R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) It is a haloalkyl or -(C3-C9)cycloalkyl.

[0053] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C7)cycloalkyl; alkyl and cycloalkyl are one, two, or three R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C7)cycloalkyl; alkyl and cycloalkyl are one or two R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) are haloalkyl or -(C3-C7)cycloalkyl; alkyl and cycloalkyl are one R Z It may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10 ,-(C1-C 12 )alkyl, -(C1-C 12 ) It is a haloalkyl or -(C3-C7)cycloalkyl.

[0054] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -OR 10That is the case.

[0055] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one, two, or three R Z It is acceptable even if it is replaced with -(C1-C 12 ) is alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one or two R Z It is acceptable even if it is replaced with -(C1-C 12 ) is alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one R Z It is acceptable even if it is replaced with -(C1-C 12 ) is alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -(C1-C 12 ) is alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is 2,2-dimethylpropyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is tert-butyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is isobutyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), one R Z The Z substituted with is cyclopropylmethyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), one R Z The Z substituted with is cyclobutylmethyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), one R Z The Z substituted with is 1-fluoro-2-methylpropyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), two R Z The substituted Z is 1,1-difluoro-2-methylpropyl.

[0056] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one, two, or three R Z It is acceptable even if it is replaced with -(C1-C 12) is a haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one or two R Z It is acceptable even if it is replaced with -(C1-C 12 ) is a haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one R Z It is acceptable even if it is replaced with -(C1-C 12 ) is a haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is -(C1-C 12 It is a haloalkyl.

[0057] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one, two, or three R Z It is a (C3-C9) cycloalkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one or two R Z It is a (C3-C9) cycloalkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one R Z It is a -(C3-C9)cycloalkyl compound which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is a -(C3-C9)cycloalkyl compound.

[0058] In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one, two, or three R Z It is a C3-C7 cycloalkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one or two R Z It is a (C3-C7) cycloalkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is one R ZIt is a -(C3-C7)cycloalkyl which may be substituted with. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is a -(C3-C7)chloroalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), Z is 2,3-dihydro-1H-inden-5-yl.

[0059] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Z These are, independently, halogen, -OR 10 , -NR 12 R 13 , or -(C1-C6)alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Z R is independently a -(C1-C6) alkyl group. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Z R is independently methyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R Z These are independently F.

[0060] In some embodiments of compounds of formula (I), (Ia), or (Ib), R 10 is H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 10 is a -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 10 is a C1-C6 alkyl or C3-C7 haloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 10 is 2,2-dimethylbutyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 10 is isopropyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 10 It is cyclohexyl.

[0061] In some embodiments of compounds of formula (I), (Ia), or (Ib), R 11 is a -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 11 These are -(C1-C6) alkyl or -(C3-C7) cycloalkyl.

[0062] In some embodiments of compounds of formula (I), (Ia), or (Ib), R 12 and R 13 R is independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), R 12 and R 13 These are independently H, -(C1-C6)alkyl, or -(C3-C7)cycloalkyl.

[0063] In some embodiments of the compounds of formula (I), (Ia), or (Ib), -XYZ is JPEG0007842812000011.jpg222169 The filename is JPEG0007842812000012.jpg177167.

[0064] In some embodiments of the compounds of formula (I), (Ia), or (Ib), -XYZ is The filename is JPEG0007842812000013.jpg154169.

[0065] In some embodiments of the compounds of formula (I), (Ia), or (Ib), -XYZ is The filename is JPEG0007842812000014.jpg91168.

[0066] In some embodiments of the compounds of formula (I), (Ia), or (Ib), -XYZ is The filename is JPEG0007842812000015.jpg30150.

[0067] In some embodiments of the compounds of formula (I), (Ia), or (Ib), -XYZ is The filename is JPEG0007842812000016.jpg28113.

[0068] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a R is independently H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a R is independently H or -(C1-C6)alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a is H. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a These are independently -(C1-C6) alkyl groups.

[0069] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a R is independently a -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R a These are independently -(C1-C6) alkyl groups.

[0070] In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R c and R d is H, -(C1-C6)alkyl, -(C1-C6)haloalkyl, or -(C3-C7)cycloalkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R c and R d R is independently H or -(C1-C6)alkyl. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each Rc and R d is H. In some embodiments of the compounds of formula (I), (Ia), or (Ib), each R c and R d These are independently -(C1-C6) alkyl groups.

[0071] In some embodiments, the compound of formula (I), (Ia), or (Ib) is selected from the compounds in Table 2 or their pharmaceutically acceptable salts, solvates, or stereoisomers. JPEG0007842812000017.jpg224167 JPEG0007842812000018.jpg220168 JPEG0007842812000019.jpg220168 JPEG0007842812000020.jpg222169 JPEG0007842812000021.jpg215170 JPEG0007842812000022.jpg208167 JPEG0007842812000023.jpg209168 JPEG0007842812000024.jpg209169 JPEG0007842812000025.jpg224167 JPEG0007842812000026.jpg230170 JPEG0007842812000027.jpg224168 JPEG0007842812000028.jpg219168 JPEG0007842812000029.jpg230170 JPEG0007842812000030.jpg220168 JPEG0007842812000031.jpg224167 JPEG0007842812000032.jpg224168 JPEG0007842812000033.jpg229169 JPEG0007842812000034.jpg222169 JPEG0007842812000035.jpg218169

[0072] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt thereof, solvate, or stereoisomer, the compound being (8S,11S,14S)-18-hydroxy-11-methyl-14-[methyl-[(2S)-2-[[4-amino-2-(4-tert-butylphenyl)-6-methylpyrimidine-5-carbonyl]amino]-3-(sulfamoylamino)propanoyl]amino]-10,13-dioxo-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0073] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt thereof, solvate, or stereoisomer, the compound being (8S,11S,14S)-18-hydroxy-11-methyl-14-[methyl-[(2S)-2-[[4-methyl-2-[4-(1-methylcyclopropyl)phenyl]pyrimidine-5-carbonyl]amino]-3-(sulfamoylamino)propanoyl]amino]-10,13-dioxo-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0074] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt thereof, solvate, or stereoisomer, the compound being (8S,11S,14S)-18-hydroxy-11-methyl-14-[methyl-[(2S)-4-amino-2-[[4-amino-2-(4-tert-butylphenyl)-6-methylpyrimidine-5-carbonyl]amino]butanoyl]amino]-10,13-dioxo-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0075] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt thereof, solvate, or stereoisomer, the compound being (8S,11S,14S)-18-hydroxy-11-methyl-14-[methyl-[(2S)-4-amino-2-[[4-methyl-2-[4-(1-methylcyclopropyl)phenyl]pyrimidine-5-carbonyl]amino]butanoyl]amino]-10,13-dioxo-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0076] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt thereof, solvate, or stereoisomer, the compound being (8S,11S,14S)-18-hydroxy-11-methyl-14-[methyl-[(2S)-4-amino-2-[[2-(4-tert-butylphenyl)-4-amino-6-difluoromethylpyrimidine-5-carbonyl]amino]butanoyl]amino]-10,13-dioxo-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0077] In some embodiments, the compound of formula (I), (Ia), or (Ib) is a compound, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, the compound being (8S,11S,14S)-14-[[(2S)-2-[[4-amino-2-(4-tert-butylphenyl)-6-(difluoromethyl)pyrimidine-5-carbonyl]amino]-3-(sulfamoylamino)propanoyl]-methyl-amino]-3,17-bis[(2R)-3-amino-2-hydroxy-propoxy]-18-hydroxy-11-methyl-10,13-dioxo-9,12-diazatricyclo[13.3.1.12,6]icosa-1(18),2(20),3,5,15(19),16-hexaene-8-carboxylic acid.

[0078] In some embodiments, the compound of formula (I), (Ia), or (Ib) is in the form of a prodrug. Some embodiments include prodrugs of the compound of formula (I), (Ia), or (Ib) that are converted to an active form in vivo via other mechanisms. In some embodiments, the compounds of the present invention are prodrugs having any of the formulas herein.

[0079] Further forms of the compounds disclosed herein Isomers / stereoisomers In some embodiments, the compounds described herein exist as geometric isomers. In some embodiments, the compounds described herein have one or more double bonds. The compounds presented herein include all cis, trans, syn, anti, entgegen (E), and tuzamen (Z) isomers, as well as corresponding mixtures thereof. In some situations, the compounds described herein have one or more chiral centers, each center existing in either an R or S configuration. The compounds described herein include all diastereomer, enantiomer, and epimer forms, as well as corresponding mixtures thereof. In further embodiments of the compounds and methods provided herein, mixtures of enantiomers and / or diastereoisomers resulting from a single preparation step, combination, or interconversion are useful for the applications described herein. In some embodiments, the compounds described herein are prepared as individual stereoisomers by reacting a racemic mixture of the compounds with an optically active resolving agent to form a pair of diastereoisomer compounds, separating the diastereomers, and recovering the optically pure enantiomers. In some embodiments, dissociable complexes are preferred. In some embodiments, the diastereomers have different physical properties (e.g., melting point, boiling point, solubility, reactivity, etc.) and are separated by utilizing these differences. In some embodiments, the diastereomers are separated by chiral chromatography, or preferably by separation / decomposition techniques based on differences in solubility. In some embodiments, the optically pure enantiomers are then recovered together with a resolving agent.

[0080] Labeled compounds In some embodiments, the compounds described herein exist in isotopically labeled forms. In some embodiments, the methods disclosed herein include methods for treating a disease by administering such isotopically labeled compounds. In some embodiments, the methods disclosed herein include methods for treating a disease by administering such isotopically labeled compounds as a pharmaceutical composition. Thus, in some embodiments, the compounds disclosed herein include isotopically labeled compounds that are identical to the compounds cited herein, except that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds described herein, or their solvates or stereoisomers, include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, for example, respectively. 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and 36 Examples include Cl. Certain isotope-labeled compounds, for example. 3 H, 14 Tritiation, i.e., the incorporation of radioactive isotopes such as 13C, is useful in drug and / or substrate tissue distribution assays. 3 H, and carbon-14, that is, 14 13C isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, deuterium, i.e., 2 Substitution with heavier isotopes such as H increases metabolic stability, for example, by extending the in vivo half-life or reducing the required dosage, resulting in specific therapeutic benefits. In some embodiments, the isotope-labeled compound or its pharmaceutically acceptable salts, solvates, or stereoisomers are prepared by any suitable method.

[0081] In some embodiments, the compounds described herein are labeled by means of other means, including but not limited to the use of chromophores or fluorescent moieties, bioluminescent labels or chemiluminescent labels.

[0082] Pharmaceutically acceptable salts In some embodiments, the compounds described herein exist as pharmaceutically acceptable salts thereof. In some embodiments, the methods disclosed herein include methods for treating a disease by administering such pharmaceutically acceptable salts. In some embodiments, the methods disclosed herein include methods for treating a disease by administering such pharmaceutically acceptable salts as a pharmaceutical composition. In some embodiments, the compounds described herein have acidic or basic groups and therefore react with a number of inorganic or organic bases, as well as both inorganic and organic acids, to form pharmaceutically acceptable salts. In some embodiments, these salts are prepared in situ during the final isolation and purification of the compounds disclosed herein, or by reacting the purified compound in its free form separately with a suitable acid or base and isolating the salts thus formed.

[0083] Examples of pharmaceutically acceptable salts include salts prepared by reacting the compounds described herein with minerals, organic acids, or inorganic bases, such as acetates, acrylates, adipicates, alginates, aspartates, benzoates, benzenesulfons, bisulfates, bisulfites, bromides, butyrates, butyn-1,4-dioate, camphorates, camphor sulfons, caproic acid, caprylates, chlorobenzoates, chlorides, citrates, cyclopentanepropionates, decanoic acid, digluconates, dihydrogen phosphate, dinitrobenzoates, dodecyl sulfates, ethanesulfons, formates, fumarates, glucoheptanoic acid, glycerophosphates, glycolates, hemisulfates, heptanoic acid, hexanoic acid, hexyn-1,6-dioate, hydroxybenzoic acid, γ-hydroxybutyrate, hydrochlorides, This includes hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate metaphosphate, methanesulfonate, methoxybenzoate, methyl benzoate, monohydrogen phosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetic acid, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, succinate, suberic acid, sebacinate, sulfonate, tartrate, thiocyanate, tosylate undeconate, and xylenesulfonate.

[0084] Furthermore, the compounds described herein include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and metaphosphoric acid, as well as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, p-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, arylsulfonic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, These can be prepared as pharmaceutically acceptable salts formed by reacting with pharmaceutically acceptable inorganic or organic acids, including but not limited to organic acids such as benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]octa-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid.

[0085] In some embodiments, the compounds described herein containing a free acid group react with a suitable base such as a pharmaceutically acceptable metal cation hydroxide, carbonate, bicarbonate, or sulfate, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, tertiary, or quaternary amine. Typical salts include alkali salts or alkaline earth salts such as lithium, sodium, potassium, calcium, and magnesium, as well as aluminum salts. Specific examples of bases include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, and N2. + (C 1-4 Examples include alkyl(4) and others.

[0086] Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, and piperazine. It should be understood that the compounds described herein also include quaternization of any basic nitrogen-containing group contained in the compound. In some embodiments, water-soluble or oil-soluble or water-dispersible or oil-dispersible products are obtained by such quaternization.

[0087] solvate In some embodiments, the compounds described herein exist as solvates. This disclosure provides a method for treating a disease by administering such solvates. This disclosure further provides a method for treating a disease by administering such solvates as pharmaceutical compositions. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and, in some embodiments, are formed during the crystallization process with a pharmaceutically acceptable solvent such as water or ethanol. When the solvent is water, a hydrate is formed, or when the solvent is alcohol, an alcoholate is formed. Solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. As just one example, hydrates of the compounds described herein can be conveniently prepared by recrystallization from an aqueous / organic solvent mixture using an organic solvent including, but not limited to, dioxane, tetrahydrofuran, or methanol. Furthermore, the compounds provided herein can exist in both non-solvated and solvated forms. Generally, the solvated form is considered equivalent to the non-solvated form in the compounds and methods provided herein.

[0088] Tautomers In some circumstances, compounds exist as tautomers. The compounds described herein include all possible tautomers within the formulas described herein. Tautomers are compounds that can be interconverted by the movement of hydrogen atoms, involving the exchange of a single bond with an adjacent double bond. In bond configurations where tautomerization is possible, a chemical equilibrium of tautomers exists. All tautomer forms of the compounds disclosed herein are intended. The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH.

[0089] In another embodiment, it is a hydrate or metabolite of any of the aforementioned compounds. In another embodiment, the pharmaceutical composition contains one of the aforementioned compounds together with a pharmaceutically acceptable excipient.

[0090] Another aspect described herein is the use of the compounds described herein in the manufacture of a drug for treating bacterial infections in patients.

[0091] In another embodiment, the method for treating a mammal in need of such treatment comprises administering to the mammal an antimicrobially effective amount of any of the above compounds at a frequency and duration sufficient to provide a beneficial effect to the mammal. In one embodiment, the mammal has a bacterial-associated infection that is resistant to treatment with arylmycin A2. In further embodiments, the causative species of bacterial infection include Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophilia, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, and Salmonella typhimurium. Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, MorganellaMorganii), Proteus mirabilis, Proteus vulgaris, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia intermedia), Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobacter pylori Campylobacter pylori, Campylobacter fetusCampylobacter fetus, Campylobacter jejuni, Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Kingella, Moraxella, Gardnerella vaginalis, Bacteroides fragilis Bacteroides fragilis, Bacteroides distasonis, Bacteroides 3452A homology group, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides splanchnicus, Clostridium difficile, Mycobacterium tuberculosis, Mycobacterium avium Mycobacterium avium), Mycobacterium intracellulare, Mycobacterium lepraeleprae), Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus It is an infectious disease involving Staphylococcus saprophyticus, Staphylococcus intermedius, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, Staphylococcus hominis, or Staphylococcus saccharolyticus.

[0092] In another embodiment, a bacterial infection is an infection involving Gram-negative bacteria. In such embodiments, Gram-negative bacteria include, for example, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumanii, Neisseria gonorrhoeae, Neisseria meningitidis, Chlamydia trachomatis, Moraxella catarrhalis, Haemophilus influenzae, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, and Helicobacter pylori. It may be Salmonella pylori, Salmonella enteritidis, Salmonella typhi, Legionella pneumophila, Haemophilus influenzae, Vibrio cholerae, Pseudomonas stutzeri, Ralstonia solanacearum, or Xylella fastidiosa.

[0093] In certain embodiments, the bacterial infection is an infection involving non-fermenting bacteria. Examples of such non-fermenting bacteria include Acinetobacter baumannii, Achromobacter xylosoxidans, Bordetella pertussis, Burkholderia cepacia (also known as Pseudomonas cepacia), Burkholderia pseudomallei (also known as Pseudomonas pseudomallei), Elizabethkingia meningoseptica (also known as Chryseobacterium meningosepticum), and Moraxella catarrhalis. It could be Branhamella catarrhalis, Pseudomonas aeruginosa, or Stenotrophomonas maltophilia.

[0094] In another embodiment, the bacterial infection is a lepB-borne infection.

[0095] In a further embodiment, the bacterial infection is an infection involving Gram-positive bacteria.

[0096] Further embodiments are methods for treating mammals in need of such treatment, comprising administering a second therapeutic agent to the mammal in addition to any of the treatment methods described above. In another embodiment, the second therapeutic agent is not an SpsB or LepB inhibitor. In another embodiment, the second therapeutic agent is an aminoglycoside antibiotic, a fluoroquinolone antibiotic, a β-lactam antibiotic, a macrolide antibiotic, a glycopeptide antibiotic, rifampicin, chloramphenicol, fluoramphenicol, colistin, mupirocin, bacitracin, daptomycin, or linezolid.

[0097] In some embodiments, a method for treating a bacterial infection in a patient, preferably a human, the treatment comprises administering a therapeutically effective or pharmacologically effective amount of a combination of 1) a β-lactam antibiotic, 2) a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, and 3) a pharmaceutically acceptable carrier. In embodiments in which the β-lactam antibiotic is used in combination with a compound disclosed herein, the β-lactam antibiotic may be a carbapenem, cephalosporin, cephamycin, monobactam, or penicillin. Examples of carbapenem antibiotics useful in the methods of the present invention include ertapenem, imipenem, biapenem, and meropenem. Examples of cephalosporin antibiotics useful in the methods of the present invention include ceftobiprole, cephthaloline, cefiprom, cefozopran, cefepime, cefotaxime, and ceftriaxone. Examples of penicillin antibiotics useful in the methods of the present invention include ampicillin, amoxacillin, piperacillin, oxacillin, cloxacillin, methicillin, and naphicillin. In some embodiments of the present invention, β-lactams may be administered together with β-lactamase inhibitors. In some embodiments of the present invention, carbapenems may be administered together with DHP inhibitors, such as cilastatin.

[0098] In various embodiments of the present invention in which the compounds and β-lactam antibiotics disclosed herein are used in combination, the β-lactam antibiotics and compounds disclosed herein may be administered sequentially or simultaneously. Preferably, the β-lactam antibiotics and compounds disclosed herein are administered together. When administered simultaneously, the β-lactam antibiotics and compounds disclosed herein may be administered in the same formulation or in separate formulations. When administered sequentially, one of the β-lactams or compounds disclosed herein may be administered first. After administration of the first compound, the other compounds may be administered, for example, within 1 to 60 minutes, for example, within 1, 2, 3, 4, 5, 10, 15, 30, or 60 minutes. In one aspect of the present invention, if a β-lactamase inhibitor is used, it may be administered separately or in a formulation with the compounds and / or β-lactam antibiotics disclosed herein. In one aspect of the present invention, when a DHP inhibitor is used to improve the stability of a carbapenem, the DHP inhibitor may be administered separately or in a formulation containing the compounds and / or carbapenem disclosed herein.

[0099] Pharmaceutical compositions comprising the compounds disclosed herein, a pharmaceutically acceptable carrier, and optionally a β-lactam antibiotic are further described herein. In embodiments in which a combination is used, the β-lactam antibiotic and the compounds disclosed herein are present in amounts such that their combination constitutes a therapeutically effective dose. Due to the enhancing effect of the compounds disclosed herein, the amount of the β-lactam antibiotic present in the combination may be less than the amount of the β-lactam antibiotic used alone. In certain embodiments, the composition further comprises a β-lactamase antibiotic.

[0100] In a further embodiment where the β-lactam antibiotic is a carbapenem, a pharmaceutical composition is provided comprising a carbapenem antibiotic, a DHP inhibitor, a compound disclosed herein, and a pharmaceutically acceptable carrier. In some embodiments where the β-lactam antibiotic is a carbapenem, the carbapenem antibiotic is preferably selected from the group consisting of ertapenem, imipenem, and meropenem.

[0101] In some embodiments, the compounds disclosed herein are for use in the treatment of bacterial infections. In some embodiments, the compounds disclosed herein are for use in the treatment of bacterial infections in combination with one or more additional therapeutic agents, including a β-lactam antibiotic. In some embodiments, the compounds disclosed herein are for use as pharmaceuticals for the treatment of bacterial infections. In some embodiments, the compounds disclosed herein are for use as pharmaceuticals for the treatment of bacterial infections in combination with one or more additional therapeutic agents, including a β-lactam antibiotic. In some embodiments, the compounds disclosed herein are for use in the preparation of pharmaceuticals for the treatment of bacterial infections. In some embodiments, the compounds disclosed herein are for use in the preparation of pharmaceuticals for the treatment of bacterial infections in combination with one or more additional therapeutic agents, including a β-lactam antibiotic.

[0102] In some embodiments described herein, the compounds disclosed herein can enhance the activity of β-lactam antimicrobial agents by inducing susceptibility to antimicrobial agents in drug-resistant strains such as MRSA. In some embodiments, the compounds disclosed herein can enhance the activity of β-lactam antimicrobial agents by reducing the dose of antimicrobial agent required for therapeutic effect in drug-susceptible strains. For example, if the compounds disclosed herein lower the minimum inhibitory concentration (MIC) of an antimicrobial agent in susceptible strains (MIC is the minimum concentration of an antimicrobial agent that completely inhibits growth), such treatment may be advantageous in order to reduce the amount of antimicrobial agent administered (which can reduce the side effects of antibiotics) or to reduce the frequency of administration. In some embodiments, the compounds disclosed herein can enhance the activity of antimicrobial agents such as carbapenems to prevent the emergence of resistant subpopulations in heterogeneous bacterial populations that include resistant subpopulations.

[0103] Enhancers can be used to enhance the activity of antimicrobial agents whose clinical efficacy is limited by the increasing prevalence of resistant bacteria. In some embodiments described herein, the compounds disclosed herein are used as enhancers and may be administered together with (simultaneously or sequentially with) β-lactam antibiotics to enable effective treatment of infections involving resistant bacteria or to reduce the amount of antimicrobial agent required to treat an infection.

[0104] In one embodiment, and this is just one example, the compounds described herein exhibit antibiotic activity useful for treating bacterial infections such as S. aureus, S. pneumoniae, E. faecalis, E. faecium, B. subtilis, and E. coli, including species resistant to many known antibiotics such as methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococcus species (VRE), multidrug-resistant E. faecium, macrolide-resistant S. aureus and S. epidermidis, and linezolid-resistant S. aureus and E. faecium.

[0105] Methicillin-resistant Staphylococcus aureus Staphylococcus aureus (S. aureus), a spherical bacterium, is the most common cause of staphylococcal infections. S. aureus is known to cause a variety of diseases, from mild skin infections such as comedones, impetigo, boils, cellulitis, folliculitis, furuncles, carbuncles, burn-like skin syndrome, and abscesses, to life-threatening diseases such as pneumonia, meningitis, osteomyelitis, endocarditis, toxic shock syndrome, and abscesses. Furthermore, S. aureus is one of the most common causes of hospital-acquired infections and frequently causes postoperative wound infections.

[0106] Methicillin was introduced in the late 1950s to treat infections caused by penicillin-resistant S. aureus. It has been previously reported that S. aureus isolates have acquired methicillin resistance (methicillin-resistant S. aureus, MRSA). The methicillin resistance gene (mecA) encodes a methicillin-resistant penicillin-binding protein that is not present in susceptible strains. mecA is located on the Staphylococcus cassette chromosome mec (SCCmec), a mobile genetic element, and four different forms with varying sizes and genetic compositions have been reported. The methicillin-resistant penicillin-binding protein enables resistance to β-lactam antibiotics, eliminating the need for their clinical use in MRSA infections.

[0107] One embodiment is a method for treating a subject having antibiotic-resistant bacteria, comprising administering to the subject a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof. In one embodiment, the bacteria are Gram-positive bacteria. In another embodiment, the Gram-positive bacteria are S. aureus. In yet another embodiment, S. aureus is resistant to or refractory to beta-lactam antibiotics. In yet another embodiment, the beta-lactam antibiotic belongs to the penicillin class. In yet another embodiment, the beta-lactam antibiotic is methicillin. In yet another embodiment, the subject has methicillin-resistant S. aureus. In one embodiment, the beta-lactam antibiotic is flucloxacillin. Another embodiment is a method for treating a subject having dicloxacillin-resistant bacteria, comprising administering to the subject a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to dicloxacillin. Furthermore, a method for treating a subject having methicillin-resistant bacteria, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject has been determined to have methicillin-resistant bacteria. In one embodiment, the subject is screened for methicillin-resistant bacteria. In another embodiment, the screening of the subject is performed by nasal culture. In a further embodiment, methicillin-resistant bacteria are detected by swabbing the subject's nostril(s) and isolating the bacteria. In another embodiment, real-time PCR and / or quantitative PCR are used to determine whether the subject has methicillin-resistant bacteria.

[0108] In one embodiment, a method for treating a subject having first-generation cephalosporin-resistant bacteria comprises administering a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to first-generation cephalosporins. In one embodiment, the bacteria are resistant to first-generation cephalosporins. In a further embodiment, the bacteria are resistant to cepacetril. In another embodiment, the bacteria are resistant to cefadroxyl. In yet another embodiment, the bacteria are resistant to cephalexin. In one embodiment, the bacteria are resistant to cephaloglysin. In another embodiment, the bacteria are resistant to cephalonium. In another embodiment, the bacteria are resistant to cephaloridine. In yet another embodiment, the bacteria are resistant to cephalothin. In a further embodiment, the bacteria are resistant to cefapyrine. In yet another embodiment, the bacteria are resistant to cefatoridine. In one embodiment, the bacteria are resistant to cefazal. In another embodiment, the bacteria are resistant to cefazedone. In yet another embodiment, the bacteria are resistant to cefazolin. In further embodiments, the bacteria are resistant to cefradin. In yet another embodiment, the bacteria are resistant to cefuroxazine. In one embodiment, the bacteria are resistant to ceftezol.

[0109] In one embodiment, a method for treating a subject having second-generation cephalosporin-resistant bacteria comprises administering a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to second-generation cephalosporins. In another embodiment, the bacteria are resistant to second-generation cephalosporins. In yet another embodiment, the bacteria are resistant to cefaclor. In yet another embodiment, the bacteria are resistant to cefonisid. In yet another embodiment, the bacteria are resistant to cefprodil. In one embodiment, the bacteria are resistant to cefuroxime. In another embodiment, the bacteria are resistant to cefzonam. In yet another embodiment, the bacteria are resistant to cefmetazole. In yet another embodiment, the bacteria are resistant to cefotetan. In yet another embodiment, the bacteria are resistant to cefoxitin.

[0110] In one embodiment, a method for treating a subject having third-generation cephalosporin-resistant bacteria comprises administering a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to third-generation cephalosporins. In another embodiment, the bacteria are resistant to third-generation cephalosporins. In yet another embodiment, the bacteria are resistant to cefcapene. In yet another embodiment, the bacteria are resistant to cefdaroxime. In yet another embodiment, the bacteria are resistant to cefdinir. In one embodiment, the bacteria are resistant to cefditoren. In another embodiment, the bacteria are resistant to cefixime. In another embodiment, the bacteria are resistant to cefmenoxime. In yet another embodiment, the bacteria are resistant to cefozidime. In yet another embodiment, the bacteria are resistant to cefotaxime. In yet another embodiment, the bacteria are resistant to cefpimizole. In one embodiment, the bacteria are resistant to cefpodoxime. In another embodiment, the bacteria are resistant to cefteram. In yet another embodiment, the bacteria are resistant to ceftibutene. In yet another embodiment, the bacteria are resistant to ceftiofur. In yet another embodiment, the bacteria are resistant to cefthiolen. In one embodiment, the bacteria are resistant to ceftizoxime. In another embodiment, the bacteria are resistant to ceftriaxone. In yet another embodiment, the bacteria are resistant to cefoperazone. In yet another embodiment, the bacteria are resistant to ceftazidime.

[0111] In one embodiment, a method for treating a subject having fourth-generation cephalosporin-resistant bacteria comprises administering a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to fourth-generation cephalosporins. In another embodiment, the bacteria are resistant to fourth-generation cephalosporins. In yet another embodiment, the bacteria are resistant to cefclizine. In yet another embodiment, the bacteria are resistant to cefepime. In yet another embodiment, the bacteria are resistant to ceffluplenum. In one embodiment, the bacteria are resistant to cefoselis. In another embodiment, the bacteria are resistant to cefozopran. In yet another embodiment, the bacteria are resistant to cefpirome. In yet another embodiment, the bacteria are refractory to cefquinome.

[0112] In one embodiment, a method for treating a subject having carbapenem-resistant bacteria comprises administering a compound disclosed herein or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein the subject is refractory to carbapenems. In another embodiment, the bacteria are resistant to carbapenems. In yet another embodiment, the bacteria are resistant to imipenems. In yet another embodiment, the bacteria are resistant to meropenems. In yet another embodiment, the bacteria are resistant to ertapenems. In one embodiment, the bacteria are resistant to faropenems. In another embodiment, the bacteria are resistant to doripenems. In another embodiment, the bacteria are resistant to panipenems. In yet another embodiment, the bacteria are resistant to biapenems.

[0113] Vancomycin-intermediate and vancomycin-resistant Staphylococcus aureus Vancomycin-intermediate resistant Staphylococcus aureus and vancomycin-resistant Staphylococcus aureus are specific types of antimicrobial-resistant Staphylococcus bacteria that are resistant to vancomycin treatment. S. aureus isolates with a vancomycin MIC of 4-8 μg / mL are classified as vancomycin-intermediate resistant, while isolates with a vancomycin MIC of 16 μg / mL or higher are classified as vancomycin-resistant (Clinical and Laboratory Standards Institute / NCCLS. Performance Standards for Antimicrobial Susceptibility Testing. Sixteenth informational supplement. M100-S16. Wayne, PA:CLSI, 2006).

[0114] As used herein, the term “minimum inhibitory concentration” (MIC) refers to the lowest concentration of an antibiotic required to inhibit the growth of a bacterial isolate in vitro. A common method for determining the MIC of an antibiotic is to prepare several tubes containing serial dilutions of the antibiotic and then inoculate them with the bacterial isolate of interest. The MIC of an antibiotic is determined from the tubes containing the minimum inhibitory concentration that are clear (no growth).

[0115] In one embodiment, a method for treating a subject having a bacterial infection comprises administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the bacterial infection includes vancomycin intermediate-resistant Staphylococcus aureus. In one embodiment, vancomycin intermediate-resistant Staphylococcus aureus bacteria have an MIC of about 4 to about 8 μg / mL. In another embodiment, vancomycin intermediate-resistant Staphylococcus aureus bacteria have an MIC of about 4 μg / mL. In yet another embodiment, vancomycin intermediate-resistant Staphylococcus aureus bacteria have an MIC of about 5 μg / mL. In yet another embodiment, vancomycin intermediate-resistant Staphylococcus aureus bacteria have an MIC of about 6 μg / mL. In yet another embodiment, vancomycin intermediate-resistant Staphylococcus aureus bacteria have an MIC of about 7 μg / mL. In one embodiment, vancomycin-intermediate resistant Staphylococcus aureus bacteria have an MIC of approximately 8 μg / mL.

[0116] In another embodiment, a method for treating a subject having a bacterial infection comprises administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the bacterial infection includes vancomycin-resistant Staphylococcus aureus. In one embodiment, vancomycin-resistant Staphylococcus aureus bacteria have an MIC between about 16 μg / mL. In another embodiment, vancomycin-resistant Staphylococcus aureus bacteria have an MIC of about 16 μg / mL or greater. In yet another embodiment, vancomycin-resistant Staphylococcus aureus bacteria have an MIC of about 20 μg / mL or greater. In yet another embodiment, vancomycin-resistant Staphylococcus aureus bacteria have an MIC of about 25 μg / mL or greater.

[0117] In one embodiment, conditions treated with the compounds described herein include, but are not limited to, endocarditis, osteomyelitis, meningitis, skin and cutaneous structure infections, genitourinary tract infections, abscesses, and necrotizing infections. In another embodiment, the compounds disclosed herein are used to treat diabetic foot infections, pressure ulcers, burn infections, animal or human bite infections, synergistic necrotizing gangrene, necrotizing fasciitis, intraperitoneal infections associated with intestinal barrier rupture (breeching), pelvic infections associated with intestinal barrier rupture, aspiration pneumonia, and postoperative wound infections. In another embodiment, the conditions described herein are caused, included in, or result of the presence of VISA and / or VRSA.

[0118] Vancomycin-resistant enterococci Enterococci are bacteria commonly found in the environment, typically residing in the human gut and female reproductive tract. These bacteria sometimes cause infections. In some cases, enterococci become resistant to vancomycin (also known as vancomycin-resistant enterococci or VRE). A common form of vancomycin resistance occurs in enterococcal strains that acquire a set of endoding genes that encode proteins instructing the peptidoglycan precursor to incorporate D-Ala-D-Lac instead of D-Ala-D-Ala. The six distinct types of vancomycin resistance exhibited by enterococci are Van-A, Van-B, Van-C, Van-D, Van-E, and Van-F. Van-A VRE may be resistant to both vancomycin and teicoplanin; in other cases, Van-B VRE is resistant to vancomycin but sensitive to teicoplanin; and in further cases, Van-C is partially resistant to vancomycin and sensitive to teicoplanin.

[0119] In one embodiment, a method for treating a subject having vancomycin-resistant enterococci, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the enterococci have developed resistance to vancomycin. In one embodiment, the subject has been previously treated with vancomycin over a long period of time. In another embodiment, the subject is hospitalized. In yet another embodiment, the subject has a weakened immune system, such as a patient in an intensive care unit or a cancer or transplant ward. In yet another embodiment, the subject has undergone a surgical procedure, such as abdominal or thoracic surgery. In yet another embodiment, the subject is colonized by VRE. In one embodiment, the subject has a medical device on which an infection has developed. In another embodiment, the medical device is a urinary catheter or a central intravenous (IV) catheter.

[0120] In another embodiment, a method for treating a subject having vancomycin-resistant enterococci, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the enterococci are resistant to Van-A.

[0121] In another embodiment, a method for treating a subject having vancomycin-resistant enterococci, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the enterococci are resistant to Van-B.

[0122] In another embodiment, a method for treating a subject having vancomycin-resistant enterococci, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to the subject, wherein the enterococci are Van-C resistant.

[0123] Administration and pharmaceutical composition The pharmaceutical compositions described herein comprise a therapeutically effective amount of one of the compounds described herein (i.e., the compounds disclosed herein) formulated with one or more pharmaceutically acceptable carriers. As used herein, the term “pharmaceutically acceptable carrier” means any kind of non-toxic, inert solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; celluloses and their derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogenic substance removal water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer. In addition, other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavorings, and fragrances, preservatives, and antioxidants may also be present in the composition at the discretion of the compounder. The pharmaceutical compositions described herein may be administered to humans and other animals orally, rectally, parenterally, intracisional, vaginally, intraperitoneally, topically (as powders, ointments, or drops, etc.), orally or as nasal sprays, or as liquid aerosols or dry powder formulations for inhalation.

[0124] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may optionally contain water, or other solvents commonly used in the art, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, as well as solubilizers, emulsifiers, and mixtures thereof. In addition to inert diluents, the oral composition may also contain humectants, emulsifiers, suspending agents, sweeteners, flavorings, and fragrances.

[0125] Preparations for injection, such as sterile aqueous or oily suspensions for injection, are optionally formulated by known techniques using appropriate dispersants or wetting agents and suspending agents. Sterile preparations for injection are optionally sterile solutions, suspensions, or emulsions for injection in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Optionally, acceptable vehicles and solvents include water, Ringer's solution, the United States Pharmacopeia, and isotonic sodium chloride solutions. In addition, sterilized non-volatile oils are conventionally employed as solvents or suspensions. For this purpose, any non-volatile oil containing synthetic monoglycerides or diglycerides can be used. In addition, fatty acids such as oleic acid are used in the preparation of injections.

[0126] Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injection media before use.

[0127] To prolong the effects of a drug, it is often desirable to delay the absorption of the drug from subcutaneous or intramuscular injection. This can be optionally achieved by using a liquid suspension of a low-water-soluble crystalline or amorphous substance. The absorption rate of the drug then depends on the dissolution rate of the drug, which may then depend on the size and morphology of the crystals. Alternatively, delayed absorption of parenterally administered drug forms can be optionally achieved by dissolving or suspending the drug in an oil vehicle. Depot formulations for injection are prepared by forming a microencapsulation matrix of the drug in a biodegradable polymer such as polylactide-polyglycolide. The drug release rate can be controlled depending on the ratio of drug to polymer and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydrous). Depot injection formulations can optionally be prepared by encapsulating the drug in liposomes or microemulsions compatible with body tissues.

[0128] Compositions for rectal or vaginal administration are preferably suppositories that can be prepared by mixing the compounds described herein (i.e., compounds disclosed herein) with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, or suppository wax, which is solid at ambient temperature but liquid at body temperature, and therefore melts in the rectum or vaginal cavity to release the active compound.

[0129] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such a solid dosage form, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate, and / or a) fillers or bulking agents such as starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as carboxymethylcellulose, arginate, gelatin, polyvinylpyrrolidinone, sucrose and gum arabic, c) water-retaining agents such as glycerol, d) agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) dissolution retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as acetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may optionally include a buffering agent.

[0130] Similar types of solid compositions can be optionally used as fillers in soft-filled and hard-filled gelatin capsules by using excipients such as lactose, i.e., milk sugar, as well as high molecular weight polyethylene glycol.

[0131] Solid dosage forms such as tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coatings and shells, such as enteric coatings and other coatings known in the pharmaceutical formulation technology. The dosage forms may optionally contain an opaque agent and may also be compositions that release only the active ingredient(s) or, preferably, a specific portion of the intestinal tract in a delayed manner. Examples of usable embedding compositions include polymers and waxes.

[0132] Similar types of solid compositions can be optionally used as fillers in soft-filled and hard-filled gelatin capsules by using excipients such as lactose, i.e., milk sugar, as well as high molecular weight polyethylene glycol.

[0133] The active compound may also be in a microencapsulated form with one or more excipients as described above. Solid dosage forms of tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings, controlled-release coatings, and other coatings known in the pharmaceutical formulation art. In such solid dosage forms, the active compound is optionally mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may optionally include additional substances other than the inert diluent, such as tableting lubricants, and other tableting aids such as magnesium stearate and microcrystalline cellulose, as is common practice. In the case of capsules, tablets, and pills, the dosage form may optionally include a buffer. The dosage form may optionally contain an opacifier and may also be a composition that releases only the active ingredient(s) or, optionally, in a delayed manner, preferably in a specific part of the intestinal tract. Examples of usable embedding compositions include polymeric substances and waxes.

[0134] Dosage forms for topical or transdermal administration of the compounds described herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed with a pharmaceutically acceptable carrier under sterile conditions and, optionally, with any necessary preservatives or buffers. Ophthalmic formulations, ear drops, and the like are also intended.

[0135] In addition to the active compounds described herein, ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0136] The compositions described herein may optionally be formulated for delivery as a liquid aerosol or an inhalable dry powder. Liquid aerosol formulations may optionally be sprayed primarily into particles that can be delivered to the terminal bronchioles and respiratory bronchioles where bacteria reside in patients with bronchial infections such as chronic bronchitis and pneumonia. Pathogenic bacteria are generally present throughout the airways, from the bronchi and bronchioles to the lung parenchyma, particularly in the terminal bronchioles and respiratory bronchioles. During exacerbations of infection, bacteria may also be present in the alveoli. Liquid aerosol and inhalable dry powder formulations are preferably delivered to the peripheral bronchi and ultimately to the parenchymal tissue.

[0137] The aerosolized formulations described herein are optionally delivered using an aerosolizing device, such as a jet, vibrating porous plate, or ultrasonic nebulizer, which is selected to enable the formation of aerosol particles having a mass medium average diameter of mainly 1 to 5 microns. Furthermore, the formulations preferably have a balanced osmotic ionic strength and chloride concentration, and a minimum aerosolizable volume that can deliver an effective dose of the compounds described herein (i.e., the compounds disclosed herein) to the site of infection. Moreover, the aerosolized formulations preferably do not negatively impair airway function and do not cause undesirable side effects.

[0138] Suitable aerosolizing devices for administering the aerosol formulations described herein include, for example, jets, vibrating porous plates, ultrasonic nebulizers, and energy-driven dry powder inhalers, which are capable of atomizing the formulation into aerosol particles mainly in the size range of 1 μm to 5 μm. In this application, “mainly” means that at least 70%, but preferably more than 90%, of all generated aerosol particles are in the range of 1 μm to 5 μm. Jet nebulizers decompose liquids into aerosol droplets using air pressure. Vibrating porous plate nebulizers operate by using a sonic vacuum generated by a rapidly vibrating porous plate to push solvent droplets through the porous plate. Ultrasonic nebulizers operate by using piezoelectric crystals to shear liquids into small aerosol droplets. For example, various suitable devices are available, such as AeroNeb® and AeroDose® vibrating porous plate nebulizers (AeroGen, Inc., Sunnyvale, California), Sidestream® nebulizers (Medic-Aid Ltd., West Sussex, England), Pari LC® and Pari LC Star® jet nebulizers (Pari Respiratory Equipment, Inc., Richmond, Virginia), and Aerosonic® (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and UltraAire® (Omron Healthcare, Inc., Vernon Hills, Illinois) ultrasonic nebulizers.

[0139] In some embodiments, the compounds described herein (i.e., the compounds disclosed herein) are formulated for use as topical powders and sprays containing, in addition to the compounds described herein, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures thereof. The sprays optionally contain conventional propellants such as chlorofluorohydrocarbons.

[0140] Transdermal patches offer the additional advantage of providing controlled delivery of compounds into the body. Such dosage forms can be manufactured by dissolving or aliquoting the compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound through the skin. This rate can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0141] According to the treatment methods described herein, bacterial infections are treated or prevented in patients such as humans or lower mammals by administering a therapeutically effective dose of the compounds described herein to the patient in the amount and time necessary to achieve the desired outcome. “Therapeutic dose” of the compounds described herein means an amount sufficient to treat a bacterial infection at a reasonable benefit / risk ratio applicable to any medical treatment. However, it should be understood that the total daily dose of the compounds and compositions described herein is to be determined by the attending physician within the bounds of sound medical judgment. The specific therapeutically effective dose level for any particular patient may depend on various factors, including the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient’s age, weight, overall health, sex, and diet; the timing, route of administration, and rate of excretion of the specific compound used; the duration of treatment; any drugs used in combination with or concurrently with the specific compound used; and similar factors known in the field of medicine.

[0142] The total daily dose of the compounds described herein (i.e., the compounds disclosed herein) administered to humans or other mammals in a single dose or in divided doses may be, for example, 0.01 to 50 mg / kg body weight or more typically 0.1 to 25 mg / kg body weight. A single-dose composition may contain such an amount or a multiple thereof to constitute a daily dose. Generally, the treatment regimens described herein involve administering to patients requiring such treatment about 10 mg to about 2000 mg of the compounds described herein per day in a single dose or in multiple doses.

[0143] Examples The compounds disclosed herein are prepared by the methods illustrated in the reaction schemes shown below. Combined with the knowledge of those skilled in the art of synthetic organic chemistry, procedures used in several embodiments to prepare the entire range of compounds disclosed and claimed herein are provided herein. General procedure: JPEG0007842812000036.jpg152164

[0144] Step 1: To a solution of compound 1 (51.0 g, 78.3 mmol) in MeOH (250 mL) and THF (250 mL), Ag2SO4 (17.1 g, 54.8 mmol) was added to I2 (21.8 g, 86.1 mmol) at 25°C. The reaction mixture was stirred at 25°C for 2 hours and filtered. The filtrate was concentrated to dryness. The residue was partitioned between ethyl acetate (300 mL) and saturated Na2S2O3 aqueous solution (300 mL). The organic phase was separated, washed with brine (2 × 300 mL), dried over Na2SO4, and concentrated to dryness to obtain crude compound 2 (60.0 g, yield 98.6%) as a yellow solid.

[0145] Step 2: To a solution of compound 2 (60.0 g, 77.2 mmol) and DIEA (38.3 mL, 231.0 mmol) in CH2Cl2 (600 mL), SEMCl (27.1 mL, 154.0 mmol) was added. The reaction mixture was stirred at 25°C for 2 hours and concentrated to dryness. The residue was diluted with ethyl acetate (500 mL), washed with water (2 × 500 mL) and brine (2 × 500 mL), dried on Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 10-40% ethyl acetate in petroleum ether) to obtain compound 3 (65.0 g, 92.8% yield) as a yellow solid.

[0146] Step 3: A mixture of compound 3 (20.00 g, 22.0 mmol), bis(pinacolate)diborone (8.39 g, 33.0 mmol), triphenylphosphine (1.16 g, 4.4 mmol), tricyclohexylphosphine (1.24 g, 4.4 mmol), Pd(OAc)2 (0.49 g, 2.2 mmol), and K2OAc (8.65 g, 88.1 mmol) in DMSO (200 mL) and water (20 mL) was stirred at 80°C for 1 hour under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (200 mL) and filtered. The filtrate was washed with water (2 × 200 mL) and brine (200 mL), dried over Na2SO4, and concentrated to obtain crude compound 4 (20.00 g, yield 97%) as a gray solid (three parallel batches were combined at this stage).

[0147] Step 4: To a solution of compound 4 (60.0 g, 66.0 mmol) in MeOH (600 mL), H2O2 (135 mL, 1.3 mol) was added. The mixture was stirred at 0°C for 2 hours and diluted with ethyl acetate (700 mL). The mixture was washed with saturated NaHCO3 aqueous solution (2 × 200 mL), saturated Na2S2O3 aqueous solution (500 mL), and brine (2 × 200 mL), dried on Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 10-50% ethyl acetate in petroleum ether) to obtain the crude product, which was further purified by preparative HPLC (water (0.04% NH3H2O ​​+ 10 mM NH4HCO3)-ACN) to obtain compound 5 (37.0 g, yield 70.1%) as a white solid.

[0148] Step 5: To a solution of compound 5 (50.0 g, 62.7 mmol) in ethanol (700 mL), 10% Pd / C (14.7 g, 13.8 mmol) and one drop of ammonia were added. The mixture was stirred at 40°C for 5 hours under hydrogen (50 psi) and filtered. The filtrate was concentrated to obtain crude compound 6 (35.0 g, yield 97.4%) as a white solid.

[0149] Step 6: To a solution of compound 6 (35.0 g, 61.0 mmol) in DMA (400 mL), CbzOSu (15.2 g, 60.9 mmol) in DMA (20.0 mL) was added dropwise at 0°C. After addition, the mixture was stirred at 15°C for 14 hours, and then diluted with ethyl acetate (500 mL). The separated organic phase was washed with brine (3 × 200 mL), dried on Na₂SO₄, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 55% ethyl acetate in petroleum ether) to obtain compound 7 (21.0 g, yield 97.0%) as a white solid.

[0150] Step 7: Compound 8 (23.1 g, 89.0 mmol) and K2CO3 (24.6 g, 178.0 mmol) were added to a solution of compound 7 (21.0 g, 29.7 mmol) in DMF (84.0 mL). The mixture was stirred at 50°C for 16 hours, and the remaining portion of compound 8 (23.1 g, 89.0 mmol) and K2CO3 (24.6 g, 178 mmol) was added. The reaction mixture was stirred at 50°C for 1 hour, diluted with ethyl acetate (500 mL), and filtered. The combined organic layer was washed with brine (2 × 300 mL), dried over Na2SO4, concentrated, and dried. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 55% ethyl acetate in petroleum ether) to obtain compound 9 (20.0 g, yield 83.3%) as a white solid.

[0151] Step 8: To a solution of compound 9 (24.0 g, 29.3 mmol) in MeCN (200 mL) and water (100 mL), NaN3 (27.3 g, 420 mmol) and CeCl3 (3.6 g, 14.6 mmol) were added. The mixture was stirred at 75°C for 16 hours. The reaction mixture was diluted with ethyl acetate (700 mL) and filtered. The combined organic layer was washed with brine (2 × 200 mL), dried over Na2SO4, concentrated and dried. The residue was dissolved in DMF (200 mL) and K2CO3 (16.2 g, 117 mmol) and MeI (12.5 g, 87.8 mmol) were added. The reaction mixture was stirred at 25°C for 1 hour, diluted with ethyl acetate (600 mL), and filtered. The combined organic layer was washed with brine (3 × 300 mL), dried over Na2SO4, concentrated and dried.

[0152] The residue was dissolved in THF (480 mL), and PPh3 (46.1 g, 176 mmol) and H2O (6.33 g, 351 mmol) were added. The solution was stirred at 35°C for 16 hours.

[0153] Boc2O (14.7 g, 67.3 mmol) was added to the above reaction mixture. The reaction mixture was stirred at 30°C for a further 1 hour and concentrated to dryness. The residue was diluted with ethyl acetate (700 mL), washed with brine (2 × 300 mL), dried on Na2SO4, and concentrated. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 17% acetone in petroleum ether, then 85% ethyl acetate in petroleum ether) to obtain compound 10 (24.4 g, yield 79.2%) as a white solid. General Procedure B: JPEG0007842812000037.jpg99164

[0154] Step 1: A mixture of compound 10 (5.00 g, 4.74 mmol) and 10% palladium carbon (1.51 g, 1.42 mmol) in ethanol (100 mL) was stirred at 35°C for 2 hours under hydrogen (50 psi) and filtered. The filtrate was concentrated to obtain crude compound 11 (4.30 g, yield 98.5%) as a white solid.

[0155] Step 2: To a solution of compound 11 (4.30 g, 4.67 mmol) and compound 12 (2.54 g, 6.08 mmol) in THF (43 mL), DIEA and HATU (2.13 g, 5.61 mmol) were added at 0°C. After addition, the reaction mixture was stirred at 25°C for 3 hours and quenched by adding methanol (1 mL). The reaction mixture was concentrated and then diluted with ethyl acetate (100 mL). This solution was washed with saturated Na2CO3 aqueous solution (150 mL) and brine (150 mL x 2), dried on Na2SO4, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 50-70% ethyl acetate in petroleum ether) to obtain compound 13 (6.00 g, 97.3% yield) as a white solid.

[0156] Step 3: To a solution of compound 13 (150 mg, 0.11 mmol) and one drop of NH3H2O ​​in ethanol (15.0 mL), 10% Pd / C (36.3 mg, 0.03 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours under an H2 atmosphere (15 psi) and filtered. The filtrate was concentrated to obtain compound 14 (135 mg, yield 63.0%) as a white solid.

[0157] The LC-MS analysis methods were as follows: LC-MS (Method 5-95 AB, ESI): ESI, the compound was eluted using a gradient from 5% AcCN / H2O to 95% AcCN / H2O over 0.7 minutes. This concentration was held for 0.4 minutes. A flow rate of 1.5 mL / min and a Merck RP-18e, 2x25 mm column were used. TFA was present at 0.05% in all chromatographic solvents; LC-MS (Method 5-100 AB, 7 min): Apparatus: Waters Acquity UPLC using a 2.1×30 mm CSH 1.8 μm C18 column held at 40°C and ESI ionization. The compound was eluted using a gradient from 5% B to 100% B in eluate A over 5.2 minutes. This concentration was held for 1.8 minutes, with a total run time of 7 minutes. The flow rate was 0.9 mL / min, and the eluates were as follows: (A) Milli-Q water + 10 mM ammonium formate (pH=3.8) and (B) MeCN. Example 1: JPEG0007842812000038.jpg69108 JPEG0007842812000039.jpg125164

[0158] Step 1: To a solution of 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (46.5 mg, 0.17 mmol) (synthesis provided in Example 17) in DMF (1.00 mL) and dichloromethane (8.00 mL), DIEA (159 μL, 0.91 mmol) and HATU (104 mg, 0.27 mmol) were added at 0°C. After 5 minutes, compound 14 (135 mg, 0.11 mmol) was added. The reaction mixture was stirred at 20°C for 2 hours and quenched by adding methanol (0.50 mL). The reaction mixture was concentrated to dryness. The residue was partitioned between ethyl acetate (30 mL) and water (20 mL). The aqueous phase was extracted with ethyl acetate (2 × 30 mL). The combined organic layers were washed with brine (3 × 80 mL), dried over Na₂SO₄, and concentrated to dryness. The crude product was purified by preparative TLC (10% methanol in dichloromethane) to obtain compound 16 (80.0 mg, 48.8% yield) as a yellow solid.

[0159] Step 5: To a solution of compound 16 (80.0 mg, 0.06 mmol) in methanol (1.00 mL), HCl (4N in methanol, 0.08 mL, 0.32 mmol) was added. The mixture was stirred at 30°C for 0.5 hours and quenched by adding NaHCO3 (70.0 mg, 0.83 mmol). The mixture was concentrated, and then THF (5 mL), water (1 mL), and Boc2O (0.01 mL, 0.06 mmol) were added. The mixture was stirred at 30°C for 0.5 hours and diluted with water (10 mL) and ethyl acetate (20 mL). The separated aqueous phase was washed with ethyl acetate (2 × 20 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, and concentrated. The residue was purified by preparative TLC (10% methanol in dichloromethane) to obtain compound 17 (50.0 mg, yield 68.7%) as a yellow solid.

[0160] Step 6: To a solution of compound 17 (50.0 mg, 0.04 mmol) in THF (4.00 mL), a solution of LiOH·H2O (4.8 mg, 0.11 mmol) in water (1 mL) was added. The reaction mixture was stirred at 20°C for 1 hour and concentrated. The residue was diluted with water (20 mL) and the pH was adjusted to 4 by adding a 5% KHSO4 aqueous solution. The mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over Na2SO4 and concentrated to obtain crude compound 18 (49.0 mg, yield 99.1%) as a white solid.

[0161] Step 7: The mixture of compound 18 (49 mg, 0.04 mmol) in 5% TFA in HFIP (3 mL, 2.01 mmol) was stirred at 35°C for 3 hours and concentrated. The residue was diluted with methanol (5 mL) and neutralized with NaHCO3. After filtration, the filtrate was purified by preparative HPLC (19-29% acetonitrile / 0.2% formic acid in water) to obtain the title compound (12.6 mg, yield 32.9%) as a white solid. 1 H NMR(400 MHz,DMSO-d6)δ(ppm)9.39(s,1 H),8.47-8.30(m,2H),7.57-7.50(m,5H),7.05-6.73(m,5H),6.36-6.22(m,2H),5.13-5.01(m ,1H),4.64(s,1H),4.22-3.99(m,10H),3.25-2.67(m,12H),1.32-1.06(m,12H).LCMS(Method 5-95 AB, ESI):R T =0.789 min,[M+H] + =995.8. Example 2: JPEG0007842812000040.jpg124160

[0162] Step 1: A solution of diethyl malonate (8.00 g, 50.00 mmol), triethyl orthoacetate (24.30 g, 150.00 mmol), ZnCl2 (2.01 mg, 0.01 mmol), and Ac2O (1.00 mL, 4.32 mmol) was heated at 135°C for 6 hours, during which additional Ac2O (1.00 mL, 4.32 mmol) was added every 30 minutes. The reaction mixture was cooled to room temperature and partitioned between (200 mL) and ethyl acetate (200 mL). The organic layer was dried over Na2SO4 and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 10-20% ethyl acetate in petroleum ether) to obtain 2-(1-ethoxyethylidene)malonate diethyl (9.00 g, yield 78.3%) as a yellow solid.

[0163] Step 2: LHMDS (1N in THF, 50.2 mL, 50.2 mmol) was added to a solution of 4-(tert-butyl)benzonitrile (4.00 g, 25.1 mmol) in THF (30 mL) at 0°C. The reaction mixture was stirred at 20°C for 16 hours. The mixture was quenched at 0°C by adding aqueous HCl (4 M, 20 mL), and then the pH was adjusted to above 8 by adding aqueous NaOH (4 M). The separated aqueous layer was extracted with dichloromethane (3 × 30 mL). The combined organic layers were dried over Na₂SO₄ and concentrated to dryness to obtain crude 4-(tert-butyl)benzimidamide (3.30 g, 74.5% yield) as a brown solid.

[0164] Step 3: Sodium (470 mg, 20.4 mmol) was added to ethanol (10 mL) and stirred for 30 minutes. This freshly prepared sodium ethoxide solution was added to a solution of 2-(1-ethoxyethylidene)malonate diethyl (4.23 g, 18.4 mmol) and 4-(tert-butyl)benzimidamide (1.80 g, 10.2 mmol) in ethanol (30 mL). The reaction mixture was stirred at 50 °C for 16 hours and quenched with saturated NH4Cl aqueous solution (30 mL). The mixture was extracted with ethyl acetate (100 mL). The combined organic layers were washed with brine (30 mL) and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum ether) to obtain 2-(4-(tert-butyl)phenyl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate ethyl (1.00 g, yield 31.1%) as a white solid.

[0165] Step 4: A mixture of POCl3 (5.00 mL, 50.6 mmol) and 2-(4-(tert-butyl)phenyl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate ethyl (1.00 g, 3.2 mmol) was stirred at 110°C for 2 hours. The mixture was concentrated under vacuum and diluted with ethyl acetate (100 mL). The solution was washed with saturated NaHCO3 aqueous solution (50 mL), dried over Na2SO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum) to obtain 2-(4-(tert-butyl)phenyl)-4-chloro-6-methylpyrimidine-5-carboxylate ethyl (0.90 g, yield 85.0%) as a pale yellow solid.

[0166] Step 5: A mixture of ethyl 2-(4-(tert-butyl)phenyl)-4-chloro-6-methylpyrimidine-5-carboxylate (1.00 g, 3.00 mmol) and ammonia (4 M in MeOH, 25.0 mL, 100.0 mmol) was stirred at 70°C for 16 hours. The reaction mixture was concentrated to dryness and diluted with ethyl acetate (100 mL). The solution was washed with water (2 × 30 mL) and brine (30 mL), dried on Na₂SO₄, and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum ether) to obtain methyl 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylate (900 mg, yield 95.6%) as a white solid.

[0167] Step 6: A mixture of methyl 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylic acid (900 mg, 3.0 mmol) and NaOH (601 mg, 15.0 mmol) in water (5 mL) and MeOH (15 mL) was stirred at 80°C for 3 hours and concentrated under reduced pressure. The residue was acidified to less than pH 5 by adding 1 M HCl and extracted with ethyl acetate (2 × 80 mL). The combined organic layer was concentrated under reduced pressure to obtain crude 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylic acid (850 mg, yield 99.1%) as a white solid.

[0168] (Formate) was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylic acid in the first step. 1H NMR(400MHz,MeOH-d4)δ(ppm)8.36-8.10(m,3H),7.63-7.34(m,2H),7.27-6.28(m,6H),5.28-5.10(m,1H),4.74-4.45 (m,2H),4.42-3.90(m,6H),3.87-3.35(m,3H),3.29-2.89(m,8H),2.87-2.37(m,3H),1.49-1.15(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.610 min,[M+H] + =1008.4. Example 3: JPEG0007842812000041.jpg145164

[0169] Step 1: A mixture of 4-hydroxybenzonitrile (5.0 g, 42.0 mmol), 1-bromo-3,3-dimethylbutane (10.4 g, 63.0 mmol), and K2CO3 (17.4 g, 126 mmol) in DMF (20 mL) was stirred at 80°C for 2 hours. After cooling to room temperature, the reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (3 × 300 mL). The combined organic layer was washed with water (2 × 300 mL) and brine (200 mL), dried over MgSO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 10% ethyl acetate in petroleum ether) to obtain 4-(3,3-dimethylbutoxy)benzonitrile (8.0 g, yield 93.8%) as a colorless oil.

[0170] Step 2: LHMDS (1N in THF, 30.0 mL, 30.0 mmol) was added to 4-(3,3-dimethylbutoxy)benzonitrile (3.0 g, 14.8 mmol) in THF (30 mL) at 0°C. The reaction mixture was stirred at 20°C for 16 hours and quenched by adding 4 M HCl until the pH was 2. Next, the mixture was adjusted to pH 12 by adding 4 M NaOH and extracted with chloroform (3 × 100 mL). The combined organic layer was dried over Na₂SO₄ and concentrated to dryness to obtain crude 4-(3,3-dimethylbutoxy)benzimidamide (3.0 g, 92.3% yield) as a yellow solid.

[0171] Step 3: To a solution of 4-(3,3-dimethylbutoxy)benzimidamide (4.70 g, 20.4 mmol) and EtONa (1.85 g, 27.2 mmol) in ethanol (20 mL), 2-(1-ethoxyethylidene)malonate (3.00 g, 13.6 mmol) was added. The reaction mixture was stirred at 50 °C for 3 hours and concentrated to dryness. The residue was partitioned between ethyl acetate (100 mL) and water (100 mL). The separated organic layer was washed with brine (50 mL), dried over MgSO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 30% ethyl acetate in petroleum ether) to obtain 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate ethyl (1.70 g, yield 34.8%) as a white solid.

[0172] Step 4: A mixture of 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate ethyl (1.70 g, 4.74 mmol) in POCl3 (30.0 mL, 325 mmol) was stirred at 110°C for 3 hours and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain 4-chloro-2-(4-(3,3-dimethylbutoxy)phenyl)-6-methylpyrimidine-5-carboxylate ethyl (1.40 g, yield 78.3%) as a white solid.

[0173] Step 5: A mixture of ethyl 4-chloro-2-(4-(3,3-dimethylbutoxy)phenyl)-6-methylpyrimidine-5-carboxylate (1.20 g, 3.18 mmol) and ammonia (4 M in MeOH, 30.0 mL, 120 mmol) was stirred at 70°C for 16 hours and concentrated to dryness. The residue was partitioned between ethyl acetate (200 mL) and water (100 mL). The separated organic layer was washed with brine (100 mL), dried over Na2SO4, and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum ether) to obtain ethyl 4-amino-2-(4-(3,3-dimethylbutoxy)phenyl)-6-pyrimidine-5-carboxylate (1.00 g, yield 87.9%) as a white solid.

[0174] Step 6: A mixture of ethyl 4-amino-2-(4-(3,3-dimethylbutoxy)phenyl)-6-methylpyrimidine-5-carboxylate (1.00 g, 2.80 mmol) and NaOH (560 mg, 14.0 mmol) in MeOH (15 mL) and water (10 mL) was stirred at 80°C for 22 hours and concentrated. The residue was adjusted to pH=5 by adding 1 M HCl. The solid was collected by filtration and dried to obtain 4-amino-2-(4-(3,3-dimethylbutoxy)phenyl)-6-methylpyrimidine-5-carboxylic acid (800 mg, yield 86.8%) as a white solid.

[0175] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(3,3-dimethylbutoxy)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.34(s,1H),8.20(d,J=8.4 Hz,2H),7.12-7.04(m,1H),6.95(d,J=8.8 Hz,2H),6.92-6.84(m 1H),6.84-6.74(m,2H),8.56(s,1H),6.44(s,1H),5.23-5.13(m,1H),4.83-4.79(m,2H),4.54-4.42(m,1H),4.26-4.20(m,1H),4.18-3 .97(m,7H),3.65-3.56(m.1H),3.40-3.33(m,4H),3.28-3.13(m,3H),3.12-2.94(m,5H),2.45(s,3H),1.80-1.68(m,3H),1.34(d,J=6.0 Hz,3H),1.01(s,9H);LCMS(Method 5-95 AB):R T =0.823 min,[M+H] + =1052.4 Example 4: JPEG0007842812000042.jpg141164

[0176] Step 1: To a solution of 4-bromophenol (7.16 g, 43.4 mmol) in DMF (30.0 mL), 1-bromo-3,3-dimethylbutane (5.00 g, 28.9 mmol) and K2CO3 (10.0 g, 72.2 mmol) were added. The reaction mixture was stirred at 50°C for 16 hours and then cooled to room temperature. The mixture was filtered, and the filtrate was diluted with ethyl acetate (80 mL) and water (80 mL). The separated organic layer was washed with brine (3 × 80 mL), dried on Na2SO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 5% methanol in dichloromethane) to obtain 1-bromo-4-(3,3-dimethylbutoxy)benzene (6.10 g, yield 82.1%) as a white solid.

[0177] Step 2: A mixture of 1-bromo-4-(3,3-dimethylbutoxy)benzene (6.10 g, 23.7 mmol), Pd(dppf)Cl2 (1.74 g, 2.37 mmol), bis(pinacolate)diborone (9.04 g, 35.6 mmol), and potassium acetate (7.00 g, 71.2 mmol) in DMF (60 mL) was heated at 80 °C for 4 hours under an N2 atmosphere, and then filtered. The filtrate was partitioned between ethyl acetate (100 mL) and water (100 mL). The separated organic phase was washed with brine (3 × 150 mL), dried over Na2SO4, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 2% ethyl acetate in petroleum ether) to obtain 2-(4-(3,3-dimethylbutoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.30 g, yield 87.3%) as a white solid.

[0178] Step 3: Ammonium acetate (83 mL, 82.7 mmol) and NaIO4 (13.3 g, 62.0 mmol) were added to a solution of 2-(4-(3,3-dimethylbutoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.30 g, 20.7 mmol) in acetone (60 mL). The reaction mixture was stirred at 40 °C for 16 hours and concentrated to dryness. The residue was diluted with water (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (50 mL), dried on Na2SO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 2% methanol in dichloromethane) to obtain (4-(3,3-dimethylbutoxy)phenyl)boronic acid (4.50 g, 20.3 mmol, 98.0% yield) as a yellow solid.

[0179] Step 4: A mixture of (4-(3,3-dimethylbutoxy)phenyl)boronic acid (200 mg, 0.90 mmol), copper diacetate (32.7 mg, 0.18 mmol), and methyl 6-oxo-1,6-dihydropyridazine-4-carboxylate (146 mg, 0.95 mmol) in dichloromethane (6 mL) and pyridine (1 mL) was stirred at 20°C for 16 hours. The reaction mixture was diluted with ethyl acetate (80 mL) and filtered. The filtrate was washed with brine (50 mL) and concentrated to dryness. The residue was separated into preparative TLC (33% ethyl acetate in petroleum ether, R f Purified by (0.3), 1-(4-(3,3-dimethylbutoxy)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylate methyl (235 mg, yield 79.0%) was obtained as a green solid.

[0180] Step 5: A mixture of methyl 1-(4-(3,3-dimethylbutoxy)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (235 mg, 0.71 mmol) and NaOH (71.1 mg, 1.78 mmol) in MeOH (10 mL) and water (3 mL) was stirred at 80°C for 1 hour. The reaction mixture was adjusted to pH=5 with HCl (1 M in water) and extracted with ethyl acetate (2 × 50 mL). The combined organic layer was dried over Na₂SO₄ and concentrated to dryness to obtain crude 1-(4-(3,3-dimethylbutoxy)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (220 mg, yield 97.8%) as a yellow solid.

[0181] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 1-(4-(3,3-dimethylbutoxy)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid. 1H NMR(400 MHz,DMSO-d6):δ(ppm)9.33(s,1H),8.46(s,1H),8.35-8.34(m,2H),8.02(s,1H),7.5 7(s,1H),7.50-7.47(m,2H),7.07-7.05(m,3H),6.90-6.66(m,8H),6.36(s,1H),6.22 (s,1H),5.05-5.02(m,2H),4.67-4.64(m,2H),4.10-3.92(m,8H),3.25-3.23(m,6H), 3.08-3.07(m,3H),2.97-2.95(m,3H),2.85-2.83(m,4H),2.68(s,1H),1.68(t,J=6.8 Hz,2H).1.16(d,J=6.4 Hz,3H),0.98(s,9H).LCMS(Method 5-95 AB,ESI),R T =0.830 min,[M+H] + =1039.3. Example 5: JPEG0007842812000043.jpg124164

[0182] Step 1: Ph3P in THF (500mL) + MeBr - (201g, 563 mmol) solution, 0 °C n-BuLi (2.5N in hexane, 225 mL, 563 mmol) was added under N2 conditions. The mixture was stirred at room temperature until the solution became clear, and then a solution of 1-(4-bromophenyl)ethane-1-one (70 g, 352 mmol) in THF (200 mL) was added dropwise. After the addition, the mixture was heated at 70°C for 20 hours and quenched by adding saturated NH4Cl aqueous solution (1000 mL). The resulting solution was extracted with ethyl acetate (2 × 1000 mL). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, petroleum ether) to obtain 1-bromo-4-(prop-1-en-2-yl)benzene (50 g, yield 72.1%) as a colorless oil.

[0183] Step 2: Add 0 to the solution of 1-bromo-4-(prop-1-en-2-yl)benzene (20.0 g, 101 mmol) in dichloromethane (100 mL). ℃ Under an N2 atmosphere, ZnEt2 (1M in toluene, 507 mL, 507 mmol) and CH2I2 (81.9 mL, 1010 mmol) were added. °C The mixture was heated for 72 hours and concentrated to dryness. The residue was partitioned between water (500 mL) and petroleum ether (500 mL). The organic layer was washed with brine (3 × 100 mL), dried over Na₂SO₄, and concentrated to dryness. The residue was then purified by preparative HPLC (55-85% acetonitrile in water / 0.225% FA) to obtain 1-bromo-4-(1-methylcyclopropyl)benzene (201 g, yield 60.7%) as a colorless oil.

[0184] Step 3: A mixture of 1-bromo-4-(1-methylcyclopropyl)benzene (28.8 g, 136 mmol), bis(pinacolate)diborone (36.3 g, 143 mmol), KOAc (40.1 g, 408 mmol), and Pd(dppf)2Cl2 (9.97 g, 13.6 mmol) in DMF (200 mL) was heated at 80°C for 6 hours under an N2 atmosphere. The reaction mixture was diluted with ethyl acetate (600 mL) and filtered. The filtrate was washed with water (2 × 200 mL) and brine (3 × 200 mL), dried over Na2SO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0%-10% ethyl acetate in petroleum ether) to obtain 4,4,5,5-tetramethyl-2-(4-(1-methylcyclopropyl)phenyl)-1,3,2-dioxaborolane (30 g, 85.2%) as a white solid.

[0185] Step 4: A mixture of 4,4,5,5-tetramethyl-2-(4-(1-methylcyclopropyl)phenyl)-1,3,2-dioxaborolane (20.0 g, 77.5 mmol), 2-chloro-4-methylpyrimidine-5-carboxylate ethyl (15.4 g, 77.5 mmol), Na2CO3 (24.6 g, 232 mmol), and Pd(dppf)2Cl2 (5.67 g, 7.75 mmol) in 1,4-dioxane (400 mL) and water (20.0 mL) was heated at 100°C for 16 hours under N2 and filtered. The filtrate was concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0%-5% ethyl acetate in petroleum) to obtain ethyl 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (26.0 g, yield 56.4%) as a white solid.

[0186] Step 5: A mixture of ethyl 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (26.0 g, 79.0 mmol) and NaOH (9.47 g, 237 mmol) in MeOH (260 mL) and water (26 mL) (26 mL) was stirred at 80°C for 16 hours and concentrated. The residue was diluted with water (100 mL) and the pH was adjusted to 4 with HCl (1 M). The mixture was filtered, and the filtrate was concentrated to dryness to obtain crude 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid (28.0 g, yield 100%) as a white solid.

[0187] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4)δ(ppm)8.86(s,1H),8.18(d,J=8.4 Hz,2H),7.32(d,J=8.4 Hz,2H),6.90-6.87(m,2H),6.78(m,2H),6.39(s,2H),5.29-5.26(m,1H),4.80-4.75(m,1H),4.60(d,J=9.2 Hz,1H),4.36-4.32(m,1H),4.27-4.19(m,2H),4.11-4.05(m,3H),3.65-3.60(m,1H),3.45-3.34(m,1H),3.2 8-3.13(m,4H),3.06(s,3H),3.00-2.86(m,1H),2.77-2.72(m,10H)2.65(m,3H),1.48(s,3H),1.37(d,J=6.8 Hz,3H),0.97-0.95(m,2H),0.89-0.88(m,2H).LCMS(Method 5-95 AB,ESI),R T =0.802 min,[M+H] + =991.2. Example 6: JPEG0007842812000044.jpg65102 JPEG0007842812000045.jpg74163

[0188] Step 1: A mixture (50 mL) of 1-bromo-4-(1-methylcyclopropyl)benzene (5.20 g, 24.6 mmol), Pd(PPh3)4 (2.80 g, 2.46 mmol), and Zn(CN)2 (6.47 g, 55.1 mmol) in DMF was heated at 120°C for 16 hours under an N2 atmosphere. After cooling to room temperature, the mixture was diluted with ethyl acetate (500 mL) and filtered. The combined organic layer was washed with brine (2 × 200 mL), dried on Na2SO4, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-0.5% ethyl acetate in petroleum ether) to obtain 4-(1-methylcyclopropyl)benzonitrile (14.0 g, yield 92.1%) as a pale yellow oil.

[0189] Step 2: A mixture of 4-(1-methylcyclopropyl)benzonitrile (5.00 g, 31.8 mmol) and LHMDS (1 N in THF, 63.6 mL, 63.6 mmol) in THF (50 mL) was stirred at 30°C for 16 hours and quenched by adding 1 M HCl (50 mL). The separated aqueous layer was adjusted to pH=8 by adding 1 M NaOH and extracted with ethyl acetate (5 × 100 mL). The combined organic layers were dried and concentrated to obtain crude 4-(1-methylcyclopropyl)benzimidamide (4.80 g, yield 86.6%) as a yellow solid.

[0190] Step 3: A mixture of 4-(1-methylcyclopropyl)benzimidamide (1.80 g, 10.3 mmol), 2-(1-ethoxyethylidene)malonate (12.0 mL, 18.6 mmol), and freshly prepared sodium ethoxide (20.7 mmol) in ethanol (40 mL) was heated at 50°C for 16 hours. After cooling to room temperature, the reaction mixture was quenched with HCl (1 M, 100 mL) and extracted with ethyl acetate (4 × 100 mL). The combined organic layer was washed with brine (100 mL), dried, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 30% ethyl acetate in petroleum ether) to obtain ethyl 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyrimidine-5-carboxylate (2.50 g, yield 77.5%) as a yellow solid.

[0191] Step 4: A mixture of ethyl 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyrimidine-5-carboxylate (6.5 g, 20.8 mmol) in POCl3 (69.3 mL, 743 mmol) was heated at 110°C for 2 hours and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 5% ethyl acetate in petroleum ether) to obtain ethyl 4-chloro-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (6.1 g, yield 88.6%) as a yellow oil.

[0192] Step 5: A mixture of ethyl 4-chloro-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (6.10 g, 18.4 mmol) in ammonia (10 N in MeOH, 60 mL, 600 mmol) was heated at 50°C for 16 hours and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 20% ethyl acetate in petroleum ether) to obtain ethyl 4-amino-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (3.00 g, yield 52.2%) as a white solid.

[0193] Step 6: A mixture of ethyl 4-amino-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid (1.85 g, 5.94 mmol) and NaOH (0.95 g, 23.8 mmol) in MeOH (20 mL) and water (10 mL) was heated at 80°C for 2 hours. After cooling to room temperature, the mixture was filtered. The filter cake was dried to obtain crude 4-amino-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid (1.68 g, yield 100%) as a white solid.

[0194] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4)δ(ppm)8.12(d,J=8.4 Hz,2H),7.47(d,J=8.4 Hz,2H),7.21-7.15(m,1H),7.02(d,J=8.4 Hz,1H),6.93-6.86(m,1H),6.85-6.80(m,1H),6.5(s,1H),6.4(s,1H),5.25-5.17(m,1H), 4.85-4.77(m,2H),4.32-4.25(m,1H),4.24-4.14(m,3H),4.12-4.04(m,2H),3.66-3.59(m 1H),3.58-3.43(m,1H),3.41-3.38(m.1H),3.37-3.31(m,2H),3.29-3.28(m,1H),3.28-3.20(m,1H),3.19-3.14(m,1H),3.13-3.07(m,2H) ),3.1(s,3H),2.75(s,1H),2.7(s,12H),2.63(s,3H),1.48(s,3H),1.42-1.28(m,3H),1.03-0.97(m,2H),0.94-0.90(m,2H).LCMS(Method 10-80 AB,ELSD),R T =1.412 min,[M+H] + =1006.7. Example 7: JPEG0007842812000046.jpg67102

[0195] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)-4,6-dimethylpyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4)δ(ppm)8.89(d,J=7.6 Hz,1H),8.55(d,J=8.8 Hz,1H),8.21(d,J=8.0 Hz,2H),7.43(d,J=8.0 Hz,2H),7.05-6.97(m,2H),6.77(s,1H),6.45(s,1H),5.43-5.39(m,1H),4.81-4.76(m,2H),4.27-4.21(m,3H),4.10-4.05(m,3H),3.63-3. 59(m,1H),3.37-3.33(m,1H),3.28-3.20(m,2H),3.12-3.07(m,6H),2.97-2.90(m,1H),2.46(s,6H),1.38(s,9H),1.37(s,3H).LCMS(Method 10-80 AB,ELSD),R T =0.652 min,[M+H] + =1007.5. Example 8: JPEG0007842812000047.jpg66104

[0196] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)-4-methylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4)δ(ppm)8.84(s,1H),8.381(s,1H),8.34-8.25(m,2H),7.55-7.47(m,1H), 7.07-6.93(m,1H),6.89-6.74(m,2H),6.66-6.56(m,1H),6.51(s,1H),5.33-5.03(m,1H) ,4.83-4.81(m,2H),4.47(s,1H),4.37-3.95(m,6H),3.65-3.50(m,1H),3.49-3.34(m,1 H),3.28-3.07(m,4H),3.07-2.94(m,4H),2.66(s,3H),1.40-1.32(m,12H) 5-95 AB,ELSD),R T =0.667 min,[M+H] +=993.4. Example 9: JPEG0007842812000048.jpg94160

[0197] Step 1: To a solution of 4-bromophenol (5.00 g, 28.9 mmol), PPh3 (22.7 g, 86.7 mmol), and cyclohexanol (8.68 g, 86.7 mmol) in THF (75.0 mL), DIAD (17.2 mL, 86.7 mmol) was slowly added at 0°C. The reaction mixture was stirred at 20°C for 3 hours and then concentrated. The residue was diluted with water (70 mL) and then extracted with ethyl acetate (3 × 60 mL). The combined organic layer was washed with brine (40 mL), dried over Na₂SO₄, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 100% petroleum ether) to obtain 1-bromo-4-(cyclohexyloxy)benzene (6.24 g, yield 85.0%) as a white solid. Next, this was converted to 1-(4-(cyclohexyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid using the same procedure as in Example 4.

[0198] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 1-(4-(cyclohexyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid. 1H NMR(400 MHz,DMSO-d6):δ(ppm)9.33(s,1H),8.44-8.34(m,2H),8.06(s,1H),7.55(d,J=1.6 HZ,1H),7.47-7.43(m,1H),7.06-7.04(m,2H),6.89-6.67(m,6H),6.36(s,1H),6.22(s 1H),5.02(s,1H),4.66(s,1H),4.40(s,1H),4.05-3.98(m,7H),3.25(s,2H),3.12-2.97(m ,6H),2.83(s,3H),1.98-1.93(m,2H),1.73-1.72(m,2H),1.53-1.15(m,9H).LCMS(Method 5-95 AB,ELSD),R T =0.815 min,[M+H] + =1037.5. Example 10: JPEG0007842812000049.jpg145164

[0199] Step 1: A mixture of compound 1 (2.50 g, 11.0 mmol) and compound 2 (4.20 g, 26.0 mmol) was stirred at 50°C for 48 hours. The reaction mixture was further heated to 70°C for 1 hour to complete the reaction. Next, the mixture was poured into ice water (100 mL) and extracted with dichloromethane (3 × 50 mL). The combined organic layer was washed with water (50 mL), dried over Na₂SO₄, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0%-5% ethyl acetate in petroleum ether) to obtain compound 3 (177 mg, yield 5.8%) as a yellow oil.

[0200] Step 2: A mixture of bis(pinacolate)diborone (177 mg, 0.70 mmol), potassium acetate (125 mg, 1.27 mmol), and compound 3 (158 mg, 0.63 mmol) in DMF (3.00 mL) was stirred at 80°C for 14 hours under an N2 atmosphere. The reaction mixture was diluted with ethyl acetate (20 mL) and filtered. The filtrate was concentrated under vacuum to obtain crude compound 4 (138 mg, 0.47 mmol) as a brown oily substance.

[0201] Step 3: Compound 4 (138 mg, 0.47 mmol), Compound 5 (102 mg, 0.51 mmol), sodium carbonate (98.8 mg, 0.93 mmol), and 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (34.1 mg, 0.05 mmol) in water (0.5 mL) and 1,4-dioxane (5 mL) were heated at 100 °C under N2 for 16 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (80 mL). The solution was washed with water (40 mL) and brine (40 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain Compound 6 (60.0 mg, yield 38.5%) as a pale oil.

[0202] Step 4: A mixture of compound 6 (60.0 mg, 0.18 mmol) and sodium hydroxide (14.4 mg, 0.36 mmol) in methanol (1.5 mL) and water (1.5 mL) was stirred at 80°C for 2 hours and concentrated. The aqueous residue was adjusted to pH=5 with 5% KHSO4 and extracted with ethyl acetate (2 × 30 mL). The combined organic layers were washed with brine (40 mL), dried, and concentrated to obtain crude compound 7 (45.8 mg, yield 83.3%) as a white solid.

[0203] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with compound 7. 1H NMR(400 MHz,MeOH-d4)δ(ppm)8.88(s,1H),8.50-8.43(m,2H),8.40(s,1H),7.58-7.52(m,2H),7.02(s,1H) ),6.89-6.79(m,2H),6.60-6.48(m,2H),5.29-5.16(m,1H),4.81-4.75(m,3H),4.46(s,1H),4.28 -3.91(m,6H),3.69-3.53(m,1H),3.43-3.35(m,1H),3.28-3.11(m,4H),3.07(s,3H),3.01-2.98( m,1H),2.81-2.60(m,4H),2.49-2.34(m,1H),1.41-1.30(m,3H),1.04-1.00(m,6H).LCMS 5-95 AB,ESI),R T =0.771 min,[M+H] + =1029.7. Example 11: JPEG0007842812000050.jpg119160

[0204] Step 1: A solution of methyl 3-aminopyrazine-2-carboxylate (5.0 g, 32.7 mmol) and m-CPBA (10.6 g, 49.0 mmol) in dichloromethane (50 mL) was stirred at 60°C for 16 hours. After cooling to room temperature, the reaction mixture was diluted with dichloromethane (250 mL) and filtered. The filtrate was concentrated to dryness, and the residue was incorporated into 10% ethyl acetate in petroleum ether (200 mL). The resulting mixture was stirred at 25°C for 1 hour and filtered. The solid was collected and dried to obtain crude 2-amino-3-(methoxycarbonyl)pyrazine 1-oxide (5.5 g, yield 99.6%) as a yellow solid.

[0205] Step 2: A mixture of 2-amino-3-(methoxycarbonyl)pyrazine 1-oxide (5.50 g, 32.5 mmol) and POCl3 (15.2 mL, 163 mmol) in DMF (30 mL) was heated at 100°C for 16 hours. After cooling to room temperature, the mixture was diluted with H2O (300 mL) and the pH was adjusted to 7 with solid NaHCO3. The resulting mixture was extracted with ethyl acetate (2 × 200 mL). The combined organic layer was washed with brine (100 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 10% ethyl acetate in petroleum ether) to obtain methyl 3-amino-5-chloropyrazine-2-carboxylate (940 mg, yield 15.4%).

[0206] Step 3: A mixture of (4-(tert-butyl)phenyl)boronic acid (937 mg, 5.26 mmol), methyl 3-amino-5-chloropyrazine-2-carboxylate (940 mg, 5.01 mmol), K3PO4 (2.08 g, 15.0 mmol), and Pd(dppf)Cl2 (367 mg, 0.50 mmol) in DMF (10.0 mL) was heated at 90°C for 16 hours under an atmosphere of N2. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (100 mL) and filtered. The combined organic layer was washed with brine (2 × 100 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 15% ethyl acetate in petroleum ether) to obtain the crude product (two isomers). The crude product was further separated by SFC to obtain methyl 3-amino-5-(4-(tert-butyl)phenyl)pyrazine-2-carboxylate (120 mg, yield 8.4%) as a yellow solid.

[0207] Step 4: A mixture of methyl 3-amino-5-(4-(tert-butyl)phenyl)pyrazine-2-carboxylate (120 mg, 0.42 mmol) and NaOH (42.1 mg, 1.05 mmol) in methanol (5 mL) and water (5 mL) (1 mL) was heated at 80°C for 16 hours. The mixture was concentrated, and the aqueous residue was adjusted to pH=2 by adding 1 M HCl. The resulting mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were dried and concentrated to obtain crude 3-amino-5-(4-(tert-butyl)phenyl)pyrazine-2-carboxylic acid (110 mg, yield 96.4%) as a yellow solid.

[0208] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 3-amino-5-(4-(tert-butyl)phenyl)pyrazine-2-carboxylic acid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.55-8.42(m,1H),8.37(br s,1H),8.09-7.91(m,2H),7.63-7.46(m,2H),7.12-6.99(m,1H),6.95-6.62(m,3H),6.48-5.93(m,2H),5.19-4.89(m,1H), 4.70-4.51(m,1H),4.32-3.85(m,5H),3.41-3.12(m,5H),3.07-2.63(m,8H),1.39-1.25(m,9H),1.24-1.04(m,3H).(Method 5-95 AB,ESI):R T =0.689 min,[M+H] + =994.4. Example 12: JPEG0007842812000051.jpg69105 JPEG0007842812000052.jpg77163

[0209] Step 1: A mixture of 4-(tert-butyl)piperidine hydrochloride (1.00 g, 5.60 mmol), 1H-pyrazole-1-carboxymidamide hydrochloride (0.82 g, 5.63 mmol), and DIEA (1.45 g, 11.3 mmol) in DMF (5.00 mL) was stirred at 60°C for 16 hours and diluted with MTBE (60 mL). The resulting suspension was stirred for 10 minutes and filtered. The solid was collected to obtain crude 4-(tert-butyl)piperidine-1-carboxymidamide (1.00 g, yield 97.0%) as a white solid.

[0210] Step 2: A mixture of 4-(tert-butyl)piperidine-1-carboxymidamide (1.00 g, 5.46 mmol), NaOEt (742 mg, 10.9 mmol), and 2-(1-ethoxyethylidene)malonate diethyl (1.88 g, 8.18 mmol) in ethanol (20.0 mL) was heated at 50°C for 20 hours and concentrated to dryness. The residue was diluted with ethyl acetate (50 mL), washed with brine (2 × 30 mL), dried, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain 2-(4-(tert-butyl)piperidine-1-yl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate ethyl (350 mg, yield 20.0%) as a pale yellow solid.

[0211] Step 3: A mixture of ethyl 2-(4-(tert-butyl)piperidine-1-yl)-4-methyl-6-oxo-1,6-dihydropyrimidine-5-carboxylate (350 mg, 1.09 mmol) and POCl3 (10 mL, 108 mmol) was heated at 90°C for 20 hours and concentrated to dryness. The residue was separated into TLC (20% ethyl acetate in petroleum ether, R) f Purified by (0.8), 2-(4-(tert-butyl)piperidine-1-yl)-4-chloro-6-methylpyrimidine-5-carboxylate ethyl (270 mg, yield 73.0%) was obtained as a pale oily substance.

[0212] Step 4: 2-(4-(tert-butyl)piperidine-1-yl)-4-chloro-6-methylpyrimidine-5-carboxylate ethyl (170 mg, 0.50 mmol) and NH3 in MeOH (5 mL) . A mixture of H2O (2.00 mL, 55.1 mmol) was heated at 60°C for 60 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (40 mL), washed with brine (20 mL), dried, and concentrated. The residue was separated into preparative TLC (10% ethyl acetate in petroleum ether, R f Purified by (0.4), 4-amino-2-(4-(tert-butyl)piperidine-1-yl)-6-methylpyrimidine-5-carboxylate ethyl (130 mg, yield 81.1%) was obtained as a pale yellow solid.

[0213] Step 5: A mixture of ethyl 4-amino-2-(4-(tert-butyl)piperidine-1-yl)-6-methylpyrimidine-5-carboxylic acid (130 mg, 0.41 mmol) and NaOH (40.6 mg, 1.01 mmol) in MeOH (5 mL) and water (1 mL) was heated at 80°C for 16 hours and concentrated. The aqueous residue was adjusted to pH=2 with 1 M HCl and extracted with ethyl acetate (3 × 10 mL). The combined organic layer was concentrated to obtain crude 4-amino-2-(4-(tert-butyl)piperidine-1-yl)-6-methylpyrimidine-5-carboxylic acid (100 mg, yield 84.3%) as a white solid.

[0214] The title compound was prepared as a white solid using the procedure described in Example 1, by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)piperidine-1-yl)-6-methylpyrimidine-5-carboxylic acid. 11H NMR (400 MHz, MeOH-d4) δ (ppm) = 8.30 (s, 1H), 7.17 - 7.14 (m, 1H), 6.95 (s, 1H), 6.89 - 6.76 (m, 2H), 6.64 - 6.54 (m, 1H), 6.50 (s, 1H), 5.12 - 5.22 (m, 1H), 4.75 - 4.71 (m, 2H), 4.55 (s, 1H), 4.28 - 4.04 (m, 5H), 3.62 - 3.57 (m, 1H), 3.41 - 3.33 (m, 2H), 3.32 - 3.35 (m, 2H), 3.21 - 3.10 (m, 2H), 3.10 - 3.04 (m, 3H), 3.01 (s, 1H), 2.94 - 2.82 (m, 1H), 2.81 - 2.74 (m, 2H), 2.74 - 2.64 (m, 1H), 2.42 - 2.33 (m, 3H), 1.85 - 1.73 (m, 2H), 1.40 - 1.28 (m, 4H), 1.26 - 1.13 (m, 2H), 0.91 (s, 9H). LCMS (Method 5 - 95 AB, ESI): R T = 0.747 min, [M / 2 + H] + = 508.5. Example 13: JPEG0007842812000053.jpg69107 JPEG0007842812000054.jpg34164

[0215] Step 1: A mixture of 4-isopropoxyphenylboronic acid (29.6 g, 164 mmol), 2-chloro-4,6-dimethylpyrimidine-5-carboxylate methyl (30.0 g, 150 mmol), Na2CO3 (31.7 g, 299 mmol), and Pd(dppf)Cl2 (10.9 g, 15.0 mmol) in water (15 mL) and 1,4-dioxane (150 mL) was heated at 100 °C for 16 hours under a nitrogen atmosphere, and then filtered. The filtrate was concentrated to dryness. The residue was partitioned between ethyl acetate (500 mL) and water (500 mL). The aqueous layer was extracted with ethyl acetate (500 mL). The combined organic layers were washed with brine (3 × 500 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum ether) to obtain 2-(4-isopropoxyphenyl)-4,6-dimethylpyrimidine-5-carboxylate methyl (42.0 g, yield 93.5%) as a white solid.

[0216] Step 2: A mixture of 2-(4-isopropoxyphenyl)-4,6-dimethylpyrimidine-5-carboxylic acid methyl (21.0 g, 69.9 mmol) and NaOH (8.39 g, 210 mmol) in MeOH (100 mL) and water (10 mL) was heated at 80°C for 22 hours and concentrated. The aqueous residue was adjusted to pH=5 by adding 1 M HCl and filtered. The collected solid was dried to obtain crude 2-(4-isopropoxyphenyl)-4,6-dimethylpyrimidine-5-carboxylic acid (19.0 g, yield 95.0%) as a white solid.

[0217] The title compound was prepared as a white solid by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-isopropoxyphenyl)-4,6-dimethylpyrimidine-5-carboxylic acid using the procedure described in Example 1. 1H NMR(400 MHz,MeOH-d4):δ(ppm)8.47(s,1H),8.30-8.28(m,2H),7.02-6.94(m,2H),6.82-6.70(m,4H),6.57(s,1H),6.43(s,1H),5.33-5.32(m,1H),4 .69-4.66(m,2H),4.35-4.09(m,6H),3.57-3.56(m,1H),3.39-3.37(m,1H),3.25-3.00(m,9H),2.54(s,6H),1.35-1.33(m,9H).LCMS(Method 5-95 AB,ESI):R T =0.608 min,[M+H] + =1009.5. Example 14: JPEG0007842812000055.jpg69107 JPEG0007842812000056.jpg32164

[0218] Step 1: 2-chloro-4-methylpyrimidine-5-carboxylate ethyl (30.0 g, 150 mmol), (4-isopropoxyphenyl)boronic acid (28.3 g, 157 mmol), sodium carbonate (31.7 g, 299 mmol), and Pd(dppf)Cl2 (10.9 g, 15.0 mmol) in water (30 mL) and 1,4-dioxane (300 mL) were heated at 100 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (300 mL), and filtered. The combined organic layer was washed with brine (100 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 2% ethyl acetate in petroleum ether) to obtain 2-(4-isopropoxyphenyl)-4-methylpyrimidine-5-carboxylate ethyl (32.5 g, yield 72.4%) as a white solid.

[0219] Step 2: A mixture of ethyl 2-(4-isopropoxyphenyl)-4-methylpyrimidine-5-carboxylic acid (32.5 g, 108 mmol) and sodium hydroxide (21.6 g, 541 mmol) in MeOH (150 mL) and water (15 mL) was stirred at 80°C for 3 hours and concentrated. The aqueous residue was adjusted to pH=5 with 2 M HCl and filtered. The filter cake was washed with water (3 × 50 mL) and dried to obtain crude 2-(4-isopropoxyphenyl)-4-methylpyrimidine-5-carboxylic acid (28.0 g, yield 95.0%) as a white solid.

[0220] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-isopropoxyphenyl)-4-methylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.82(br s,1H),8.35(d,J=8.8 Hz,2H),7.11(d,J=8.4 Hz,1H),6.99(d,J=8.8 Hz,2H),6.90(d,J=8.0 Hz,1H),6.86-6.76(m,2H),6.59(br s,1H),6.45(s,1H),5.27-5.16(m,1H),4.75-4.66(m,2H),4.65-4.45(m,2H),4.37-4.19(m,1H),4.17-4.00(m,5H),3.71-3.5 4(m,1H),3.50-3.44(m,1H),3.42-3.35(m,1H),3.29-3.09(m,5H),3.06(s,3H),2.69(s,3H),1.33-1.38(m,9H).LCMS(Method 5-95 AB,ESI):R T =0.609 min,[M+H] + =996.7. Example 15: JPEG0007842812000057.jpg67102 JPEG0007842812000058.jpg39164

[0221] Step 1: 4,4,5,5-tetramethyl-2-(4-(1-methylcyclopropyl)phenyl)-1,3,2-dioxaborolane (4.67 g, 18.1 mmol), a mixture of 2-chloro-4-methylpyrimidine ethyl-5-carboxylate (3.30 g, 16.5 mmol), sodium carbonate (3.49 g, 32.9 mmol), and Pd(dppf)Cl in water (3 mL) and 1,4-dioxane (30 mL). 2( A mixture of 1.20 g and 1.64 mmol of the compound was heated under a nitrogen atmosphere at 100°C for 16 hours. The reaction mixture was cooled to room temperature, the reaction mixture was diluted with ethyl acetate (300 mL), and filtered. The filtrate was washed with water (100 mL) and brine (50 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain ethyl 4,6-dimethyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylate (2.10 g, yield 43.1%) as a white solid.

[0222] Step 2: A mixture of ethyl 4,6-dimethyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid (2.10 g, 7.09 mmol) and sodium hydroxide (567 mg, 14.2 mmol) in MeOH (30 mL) and water (3 mL) was stirred at 80°C for 4 hours and concentrated. The aqueous residue was adjusted to pH=5 with 1 M HCl and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried, and concentrated. The crude product was recrystallized with 10% ethyl acetate in petroleum ether (10 mL) to obtain 4,6-dimethyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid (1.10 g, 55% yield) as a white solid.

[0223] The title compound was prepared using the procedure of Example 1, by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4,6-dimethyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid. 1H NMR(400 MHz,DMSO-d6):δ(ppm)8.27(d,J=8.4H,2H),7.34(d,J=8.0 Hz,2H),7.17-7.06(m,1H),7.04-6.90(m,1H),6.87-6.70(m,2H),6.44(s,1H),6.30(m,1H),5.39-5.05(m,1H),4.75-4.58(m,1H),4. 48-4.35(m,1H),4.18-3.90(m,6H),3.31-3.13(m,3H),3.06-2.95(m,5H),2.93-2.64(m,3H),2.44(s,6H),1.42(s,3H),1.21(d,J=6.4 Hz,3H),0.93-0.86(m,2H),0.85-0.77(m,2H).LCMS(Method 5-95 AB,ESI):R T =0.631 min,[M+H] + =1006.3. Example 16: JPEG0007842812000059.jpg148160

[0224] Step 1: To a solution of 4-bromophenol (14.3 g, 86.7 mmol) in DMF (100 mL), 1-bromo-3,3-dimethylbutane (10.0 g, 57.8 mmol) and potassium carbonate (20.0 g, 145 mmol) were added. The reaction mixture was stirred at 50°C for 16 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (500 mL), and filtered. The combined organic layer was washed with brine (400 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-2% ethyl acetate in petroleum ether) to obtain 1-bromo-4-(3,3-dimethylbutoxy)benzene (14.5 g, yield 97.5%) as a white solid.

[0225] Step 2: 1-Bromo-4-(3,3-dimethylbutoxy)benzene (10.0 g, 38.9 mmol), bis(pinacolate)diborone (10.4 g, 40.8 mmol), potassium acetate (11.5 g, 116.7 mmol), and Pd(dppf)2Cl2 (2.85 g, 3.89 mmol) in DMF (100 mL) were heated at 80°C for 16 hours under a nitrogen atmosphere and filtered. The filtered mixture was diluted with water (500 mL) and extracted with ethyl acetate (3 × 500 mL). The combined organic layers were washed with brine (500 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 2% ethyl acetate in petroleum ether) to obtain 2-(4-(3,3-dimethylbutoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (16.0 g, 52.6 mmol) as a colorless oil.

[0226] Step 3: A mixture of 2-(4-(3,3-dimethylbutoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.00 g, 3.29 mmol), 2-chloro-4-methylpyrimidine-5-carboxylate ethyl (692 mg, 3.45 mmol), sodium carbonate (697 mg, 6.57 mmol), and Pd(dppf)Cl2 (240 mg, 0.33 mmol) in water (1 mL) and 1,4-dioxane (10 mL) was heated at 100 °C for 16 hours under a nitrogen atmosphere and diluted with ethyl acetate (100 mL). The resulting mixture was filtered, the filtrate was washed with water (2 × 40 mL) and brine (50 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-2% ethyl acetate in petroleum ether) to obtain 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methylpyrimidine-5-carboxylate ethyl (460 mg, yield 40.9%) as a colorless oil.

[0227] Step 4: A mixture of ethyl 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methylpyrimidine-5-carboxylic acid (460 mg, 1.34 mmol) and sodium hydroxide (269 mg, 6.72 mmol) in MeOH (10 mL) and water (10 mL) was stirred at 80°C for 16 hours and concentrated. The aqueous residue was adjusted to pH=5 with 1 M HCl and filtered. The filter cake was dried to obtain crude 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methylpyrimidine-5-carboxylic acid (400 mg, yield 94.7%) as a white solid.

[0228] The title compound was prepared using the procedure of Example 1, by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.80(s,1H),8.27(d,J=8.8 Hz,2H),7.06-7.05(m,1H),6.93(d,J=8.8 Hz,2H),6.85-6.86(m,1H),6.69-6.66(m,2H),6.49(s,1H),6.37(s,1H),5.23-5.21(m,1H),4.55-4.50(m,2H),4.23-4.05(m,1H),3.60- 3.58(m,1H),3.50-3.38(m,1H),3.26-3.24(m,2H),3.17-3.11(m,2H),3.18-2.99(m,4H),2.64(s,3H),1.76-1.70(m,2H),1.35(d,J=6.4 Hz,3H),0.99(s,9H).LCMS(Method 5-95 AB,ESI):R T =0.844 min,[M+H] + =1037.4. Example 17: JPEG0007842812000060.jpg64108 JPEG0007842812000061.jpg117161

[0229] Step 1: Compound 10 (200 mg, 0.19 mmol) and 1 drop of NH3 in ethanol (15 mL). . To a solution of H2O, 10% Pd / C (60.6 mg, 0.06 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours under an H2 atmosphere (15 psi) and filtered. The filtrate was concentrated to obtain crude compound 11 (174 mg, 100% yield) as a white solid.

[0230] Step 2: To a solution of compound 11 (174 mg, 0.19 mmol) in dichloromethane (10 mL), DIEA (0.13 mL, 0.76 mmol) and (S)-2-(((benzyloxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)butanoic acid (134 mg, 0.38 mmol), followed by a solution of HATU (86.8 mg, 0.23 mmol) in DMF (1 mL) were added at 0°C. The mixture was stirred at 20°C for 2 hours and quenched with MeOH (0.5 mL). The mixture was diluted with water (10 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine (3 × 30 mL), dried, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 5% MeOH in dichloromethane) to obtain compound 19 (200 mg, yield 83.8%) as a white solid.

[0231] Step 3: To a solution of compound 19 (200 mg, 0.16 mmol) and one drop of ammonium hydroxide in ethanol (15 mL), 10% Pd / C (76.4 mg, 0.07 mmol) was added. The reaction mixture was stirred at 40°C for 6 hours under an H2 atmosphere (15 psi) and filtered. The filtrate was concentrated to obtain crude compound 20 (178 mg, 100% yield) as a white solid. JPEG0007842812000062.jpg75160

[0232] Step 1: A mixture of 6-oxo-1,6-dihydropyridazine-4-carboxylic acid (2.00 g, 14.3 mmol) and HCl (4 M in MeOH, 15.0 mL, 60.0 mmol) was stirred at 25°C for 24 hours and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 30-60% ethyl acetate in petroleum ether) to obtain methyl 6-oxo-1,6-dihydropyridazine-4-carboxylic acid (1.00 g, yield 45.5%) as a white solid.

[0233] Step 2: A mixture of (4-(tert-butyl)phenyl)boronic acid (118 mg, 0.65 mmol), 6-oxo-1,6-dihydropyridazine-4-carboxylate methyl (500 mg, 3.24 mmol), copper(II) acetate (24 mg, 0.13 mmol) and pyridine (2 mL) in dichloromethane (10 mL) was stirred at 25°C for 24 hours and concentrated. The residue was diluted with ethyl acetate (80 mL), washed with brine (2 × 30 mL), dried, and concentrated. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 33% ethyl acetate in petroleum ether) to obtain 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylate methyl (770 mg, yield 82.9%) as a white solid.

[0234] Step 3: A mixture of methyl 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (770 mg, 2.69 mmol) and lithium hydroxide hydrate (247 mg, 10.8 mmol) in THF (10 mL) and water (2 mL) was stirred at 25°C for 2 hours and concentrated. The residue was adjusted to pH=3 with 1 M HCl and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were dried and concentrated to obtain crude 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (730 mg, yield 99.7%) as a white solid.

[0235] The title compound was prepared using the procedure of Example 1, with compound 14 replaced by compound 20.1 H NMR(400 MHz,DMSO-d6):δ(ppm)8.43(s,1H),8.38-8.28(m,2H),8.14-8.07(m,1H),7.60-7.44(m,5H),7.10-7 .04(m,1H),6.93-6.91(m,1H),6.78(s,2H),6.35(s,1H),6.24(s,1H),5.05-4.94(m,1H),4.67-4.65 (m,3H),4.35(s,1H),4.12-3.93(m,11H),3.3-3.27(m,2H),3.17-3.10(m,3H),2.99-2.97(m,4H),2. 80(s,4H),2.67-2.64(m,1H),2.12-2.03(m,2H),1.33-1.28(m,9H),1.17-1.00(m,3H).LCMS(Method 5-95 AB,ESI):R T =0.797 min,[M+H] + =929.4. Example 18: JPEG0007842812000063.jpg64102

[0236] The title compound was prepared from compound 20 using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(isopentyloxy)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4)δ(ppm)8.41(s,1H),8.23(d,J=8.8 Hz,2H),7.18-7.04(m,1H),6.97(d,J=8.8 Hz,2H),6.93-6.87(m,1H),6.84(s,1H),6.81(s,1H),6.60(s,1H),6.46(s,1H),4.87-4.73(m,2H),4. 52-4.41(m,1H),4.24-4.01(m,8H),3.27-2.97(m,11H),2.45(s,3H),2.31-2.11(m,2H),1.75(t,J=6.8 Hz,2H),1.34(d,J=6.8 Hz,3H),1.02(s,9H).LCMS(Method 5-95 AB,ESI):RT =0.767 min,[M+H] + =987.8. Example 19: JPEG0007842812000064.jpg64104

[0237] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)-4,6-dimethylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4)δ(ppm)8.34(s,1H),8.28(d,J=8.4 Hz,2H),7.48(d,J=8.8 Hz,2H),7.02(d,J=8.0 Hz,1H),6.89(d,J=8.8 Hz,1H),6.81-6.80(m,1H),6.66(s,1H),6.55(s,1H),6.56(s,1H),5.21- 5.18(m,1H),4.81-4.76(m,1H),4.41-4.39(m,1H),4.24-4.06(m,6H),3. 27-3.22(m,2H),3.19-3.12(m,4H),3.10-3.07(m,1H),3.03(s,3H),2.99 -2.95(m,1H),2.51(s,6H),2.30-2.17(m,2H),1.37(s,9H),1.34(d,J=6.8 Hz,3H).LCMS(Method 5-95 AB,ESI):R T =0.622 min,[M+H] + =942.4. Example 20: JPEG0007842812000065.jpg61102

[0238] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-isopropoxyphenyl)-4-methylpyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4):δ(ppm)8.89(s,1H),8.23(d,J=8.8 Hz,2H),7.05-6.92(m,4H),6.87-6.82(m,1 H),6.66(s,1H),6.51(s,1H),6.42(s,1 H),5.20-5.11(m,1H),4.83-4.72(m,2 H),4.69-4.59(m,1H),4.40-4.03(m,6 H),3.40-3.32(m,1H),3.29-3.07(m,6H),2.99(s,3H),2.97-2.80(m,1H),2. 70(s,12H),2.68(s,3H),2.39-2.15(m,2H),1.47-1.30(m,9H).LCMS(Method 5-95 AB,ESI):R T =0.732 min,[M+H] + =930.7. Example 21: JPEG0007842812000066.jpg61105

[0239] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-6-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.36(s,1H),8.18-8.15(m,2H),7.33-7.31(m,2H ),7.11(s,1H),6.93-6.91(m,1H),6.86-6.81(m,2H),6.58-6.46(m,3H), 5.04-5.03(m,1H),4.82(s,2H),4.54(s,1H),4.22-4.05(m,6H),3.18-2 .98(m,10H),2.46(s,3H),2.25-2.18(m,2H),1.44(s,3H),1.34(d,J=6.4 HZ,2H),0.93-0.91(m,2H),0.82-0.80(m,2H).LCMS(Method 5-95 AB,ESI):R T =0.58 min,[M+H] + =941.5. Example 22: JPEG0007842812000067.jpg61102

[0240] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 1-(4-(3,3-dimethylbutoxy)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid. 1 1H NMR (400 MHz,MeOH-d4):δ(ppm)8.08(s,1H),7.45-6.85(m,9H),6.60(s,1H),6.31(s,1H) ),5.03-5.01(m,1H),4.74-4.71(m,1H),4.37(s,1H),4.25(s,1H),4.11-4.06(m ,4H),3.91-3.90(m,1H),3.38-3.31(m,3H),3.30-3.12(m,3H),2.99-2.90(m,7H ),2.18(s,2H),1.76-1.73(m,2H),1.37-1.35(m,3H),1.02(s,2H).LCMS(Method 5-95 AB,ESI):R T =0.660 min,[M+H] + =974.6. Example 23: JPEG0007842812000068.jpg61105

[0241] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)-4-methylpyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4):δ(ppm)8.72(s,1H),8.34(m,1H),8.26-8.24(d,J=8.0Hz,1H), 7.47-7.45(d,J=8.0Hz,2H),6.92-6.91(d,J=4.0Hz,1H),6.81-6.79(d,J=8.0 Hz,2H),6.61(s,1H),6.47(s,2H),5.10-5.09(m,1H),4.80-4.76(m,2H),4.48-4.46(m,1H),4.29-4.27(d,J=8.0 Hz,1H),4.18-4.12(m,4H),4.03-4.02(d,J=4.0Hz,1H),3.26(s,1H),3.20-3.15(m,4H),3.10-3.14(m,1H),2.97(s,3H), 2.92-2.89(d,J=12.0Hz,1H),2.76-2.67(m,1H),2.62-2.60(m,3H),2.29-2.23(m,3H),1.35-1.32(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.769 min,[M+H] + =928.8. Example 24: JPEG0007842812000069.jpg62104

[0242] The title compound was prepared from compound 20 using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylic acid. 11H NMR (400 MHz, MeOH-d4): δ (ppm) 8.24 (s, 1H), 8.23 - 8.17 (m, 2H), 7.54 - 7.48 (m, 1H), 7.21 - 7.14 (m, 1H), 7.02 - 6.92 (m, 1H), 6.88 (s, 1H), 6.85 (s, 1H), 6.59 (s, 1H), 6.45 (s, 1H), 5.30 - 5.01 (m, 1H), 4.83 - 4.79 (m, 2H), 4.66 - 4.52 (m, 1H), April 25, 2023 4.25 - 4.04 (m, 6H), 3.25 - 2.99 (m, 10H), 2.52 - 2.43 (m, 3H), 2.33 - 2.08 (m, 2H), 1.40 - 1.33 (s, 12H). LCMS (Method 5-95 AB, ESI): R T = 0.592 min, [M + H] + = 943.4. Example 25: JPEG0007842812000070.jpg61107

[0243] The title compound was prepared from Compound 20 using the procedure of Example 1, replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)piperidin-1-yl)-6-methylpyrimidine-5-carboxylic acid. 1 1H NMR (400 MHz, MeOH-d4): δ (ppm) 7.27 - 7.21 (m, 1H), 7.04 (d, J = 8 Hz, 1H), 6.92 - 6.87 (m, 2H), 6.55 (m, 1H), 6.38 (s, 1H), 5.04 - 5.01 (m, 1H), 4.72 (m, 2H), 4.28 - 4.07 (m, 6H), 3.52 - 3.34 (m, 2H), 3.31 - 3.04 (m, 8H), 2.99 (s, 3H), 2.95 - 2.86 (m, 2H), 2.40 (s, 3H), 2.28 - 2.12 (m, October 25, 2023 2H), 1.85 (m, 2H), 1.44 - 1.33 (m, 4H), 1.30 - 1.19 (m, 2H), 0.92 (s, 9H). LCMS (Method 5-95 AB, ESI): R T = 0.731 min, [M / 2 + H] + = 479.5. Example 26: JPEG0007842812000071.jpg64107

[0244] Compound 20 was prepared from the title compound using the procedure of Example 1, by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 1-(4-(cyclohexyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.34-8.14(m,1H),7.48-7.33(m,1H),7.19-6.62(m, 9H),6.35-6.30(m,1H),5.01-4.90(m,1H),4.73-4.71(m,1H),4.37-3.92(m, 7H),3.36-3.30(m,4H),3.18-2.91(m,9H),2.28-2.18(m,2H),2.18-1.98(m ,2H),1.82-1.80(m,2H),1.55-1.43(m,6H),1.36-1.35(m,3H).LCMS(Method 5-95 AB,ESI):R T =0.696 min,[M / 2+H] + =485.9. Example 27: JPEG0007842812000072.jpg66107

[0245] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(3,3-dimethylbutoxy)phenyl)-4-methylpyrimidine-5-carboxylic acid. 11H NMR (400 MHz, MeOH-d4): δ (ppm) 8.79 (s, 1H), 8.37 (d, J = 8.4 Hz, 2H), 7.07 - 7.00 (m, 3H), 6.87 - 6.83 (m, 2H), 6.72 (s, 1H), 6.58 (s, 1H), 6.47 (s, 1H), 4.84 - 4.83 (m, 2H), 4.41 (s, 1H), 4.27 - 4.03 (m, 8H), 3.32 - 2.99 (m, 11H), 2.74 - 2.68 (m, 3H), 2.32 - 2.19 (m, 2H), 1.78 - 1.75 (m, 2H), 1.39 - 1.34 (m, 3H), 1.04 (s, 9H). LCMS (Method 5-95 AB, ESI): R T = 0.825 min, [M+H] + = 973.0. Example 28: JPEG0007842812000073.jpg64107

[0246] The title compound was prepared from Compound 20 using the procedure of Example 1, replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid. 1 1H NMR (400 MHz, MeOH-d4): δ (ppm) 8.79 (s, 1H), 8.20 (d, J = 8.4 Hz, 2H), 7.31 (d, J = 8.4 Hz, 2H), 7.00 - 6.80 (m, 3H), 6.88 (s, 1H), 6.44 (s, 1H), 6.40 (s, 1H), 5.18 - 5.10 (m, 1H), 4.84 - 4.71 (m, 1H), 4.65 - 4.53 (m, 1H), 4.37 - 4.30 (m, 1H), 4.28 - 4.17 (m, 2H), 4.16 - 3.97 (m, 3H), 3.27 - 3.08 (m, 6H), 3.06 - 2.73 (m, 5H), 2.70 (s, 12H), 2.62 (s, 3H), 2.37 - 2.15 (m, 2H), 1.47 (s, 3H), 1.37 (d, J = 6.8 Hz, 3H), 1.02 - 0.92 (m, 2H), 0.91 - 0.82 (m, 2H). LCMS (Method 5-95 AB, ESI): R T = 0.763 min, [M+H]+ =926.3. Example 29: JPEG0007842812000074.jpg62105 JPEG0007842812000075.jpg75164

[0247] Step 1: A mixture of 4,4,5,5-tetramethyl-2-(4-(1-methylcyclopropyl)phenyl)-1,3,2-dioxaborolane (8.0 g, 31.0 mmol), sodium periodate (33.1 g, 155 mmol), and ammonium acetate (11.9 g, 155 mmol) in acetone (80 mL) and water (80 mL) was stirred at 25°C for 16 hours and filtered. The filtrate was concentrated, and the aqueous residue was extracted with ethyl acetate (2 × 150 mL). The combined organic layer was concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-25% ethyl acetate in petroleum ether) to obtain (4-(1-methylcyclopropyl)phenyl)boronic acid (5.4 g, yield 99.0%) as a white solid.

[0248] Step 2: A mixture of (4-(1-methylcyclopropyl)phenyl)boronic acid (1.00 g, 5.68 mmol), copper(II) acetate (0.21 g, 1.14 mmol), and methyl 6-oxo-1,6-dihydropyridazine-4-carboxylate (0.92 g, 5.97 mmol) in dichloromethane (10 mL) and pyridine (1 mL) was stirred at 25°C for 16 hours. The mixture was diluted with 1 M HCl (50 mL) and then extracted with dichloromethane (2 × 100 mL). The combined organic layer was washed with brine (100 mL), dried, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-25% ethyl acetate in petroleum ether) to obtain 1-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylate methyl (1.00 g, 61.7%) as a yellow solid.

[0249] Step 3: A mixture of methyl 1-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (6.0 g, 21.1 mmol) and sodium hydroxide (2.1 g, 52.76 mmol) in MeOH (50 mL) and water (20 mL) was heated at 80°C for 2 hours and concentrated. The aqueous residue was adjusted to pH=5 with 1N HCl and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (200 mL), dried, and concentrated to obtain crude 1-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid (5.30 g, yield 92.9%) as a pale yellow solid.

[0250] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 1-(4-(1-methylcyclopropyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.32(d,J=2.0 Hz,1H),7.53(d,J=2.0 Hz,1H),7.39-7.32(m,4H),7.15-7.13(m,1H),7.02-7.00(m,1H),6.90( s,1H),6.68(s,1H),6.50(s,1H),6.41(s,1H),4.87-4.78(m,2H),4.36- 4.34(m,1H),4.20-4.04(m,6H),3.37-3.30(m,3H),3.21-3.05(m,5H),2 .87(s,3H),2.71(s,12H),3.33-3.21(m,2H),1.43(s,3H),1.37(d,J=6.8 Hz,3H),0.91-0.88(m,2H),0.81-0.78(m,2H).LCMS(Method 5-95 AB,ESI):R T =0.739 min,[M+H] + =928.4. Example 30: JPEG0007842812000076.jpg56107

[0251] The title compound was prepared using the procedure of Example 22, with (S)-2-(((benzyloxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)butanoic acid replaced with (S)-2-(((benzyloxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid, and then prepared using general procedure B, with 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid replaced with 2-(4-(tert-butyl)phenyl)-4,6-dimethylpyrimidine-5-carboxylic acid. 1 H NMR(400 MHz,MeOH-d4):δ(ppm)8.43(s,1H),8.33(d,J=8.8 Hz,2H),7.53(d,J=8.8 Hz,2H),7.07(d,J=8.0 Hz,1H),6.88(d,J=8.4 Hz,1H),6.83(s,1H),6.76(s,1H),6.61(s,1H),6.39(s,1H),5.33-5.30(m,1H),4.38-4.36(m,2H), 4.21-4.03(m,2H),3.49-3.44(m,6H),3.26-2.96(m,10H),2.57(s,6H),1.38(s,9H),1.34(d,J=6.8 Hz,3H).LCMS(Method 5-95 AB,ESI),R T =0.611 min,[M+H] + =928.5. Example 31: JPEG0007842812000077.jpg56104

[0252] The title compound was prepared using the procedure of Example 22, with (S)-2-(((benzyloxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)butanoic acid replaced with (S)-2-(((benzyloxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid, and then using the general procedure B, with 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid replaced with 2-(4-(tert-butyl)phenyl)-4-methylpyrimidine-5-carboxylic acid. 1H NMR(400 MHz,MeOH-d4):δ(ppm)8.91(s,1H),8.35(d,J=8.4 Hz,2H),7.54(d,J=8.8 Hz,2H),7.08-7.00(m,1H),6.96-6.80(m,2H),6.71-6.60(m,1H),6.54 (s,1H),6.41-6.27(m,1H),5.50-5.25(m,1H),4.76-4.58(m,1H),4.54- 4.41(m,1H),4.26-3.95(m,6H),3.57-3.48(m,1H),3.41-3.35(m,1H),3.28-2.84(m,9H),2.71-2.66(m,3H),1.42-1.30(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.770 min,[M+H] + =914.8. Example 32: JPEG0007842812000078.jpg134164

[0253] The general procedure for the synthesis of 4-amino-2-(4-tert-butylphenyl)-6-(difluoromethyl)pyrimidine-5-carbonitrile was followed using the procedure described in Chem.Eur.J.2018,24,1311-1316.

[0254] Step 1: A solution of boron trifluoride diethyl etherate (2.68 mL, 21.71 mmol, 1.05 equivalents) in acetonitrile (19.2 mL) was stirred at -30°C under N2 for 15 minutes. 1,1,2,2-tetrafluoro-N,N-dimethylethanamine (3.0 g, 20.68 mmol, 1.0 equivalent) in acetonitrile (27 mL) was added, and the mixture was stirred for 5 minutes. The reaction mixture was allowed to return to room temperature. A solution of malononitrile (1.37 g, 20.68 mmol, 1.0 equivalent) in dry acetonitrile (19.2 mL) was added to the solution under an inert atmosphere at 25°C, followed by the addition of N,N-diisopropylethylamine (5.4 mL, 31.01 mmol, 1.5 equivalents). The mixture was stirred for 2 hours. The reaction was quenched by the addition of silica gel. The solution was concentrated under reduced pressure to obtain a yellow solid residue, which was purified by column chromatography (silica gel, 100-200 mesh, 10-50% siRNA in heptane) to obtain 2-[1-(dimethylamino)-2,2-difluoroethylidene]propanedinitrile (1.68 g, 9.816 mmol, yield 48%) as a yellow solid.

[0255] Step 2: 2-[1-(dimethylamino)-2,2-difluoroethylidene]propanedinitrile (150 mg, 0.8800 mmol, 1.0 equivalent) and (4-tert-butylbenzenecarboximidoyl)ammonium chloride (372.87 mg, 1.75 mmol, 2.0 equivalents) were dissolved in anhydrous ethanol (2.9214 mL, 0.3 M) and placed in a microwave vial, which was then capped. The reaction mixture was heated to 70°C under microwave irradiation and stirred for 30 minutes. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 0-50% ethyl phosphate in heptane) to obtain 4-amino-2-(4-tert-butylphenyl)-6-(difluoromethyl)pyrimidine-5-carbonitrile (175 mg, 0.5789 mmol, yield 66%) as a white solid.

[0256] Step 3: 160 mg (0.5300 mmol, 1.0 equivalent) of 4-amino-2-(4-tert-butylphenyl)-6-(difluoromethyl)pyrimidine-5-carbonitrile was dissolved in 2.6 mL of anhydrous EtOH. 0.66 mL (2.65 mmol, 5.0 equivalent) of 4 M potassium hydroxide solution was added, and the reaction mixture was heated to 70°C for 18 hours. The reaction mixture was cooled to room temperature, a 1 N solution of KHSO4 was added, and the reaction mixture was diluted with ELISA (20 mL). The phases were separated, and the aqueous layer was extracted with ELISA (2 × 20 mL). The organic layers were combined, washed with brine (2 × 20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain 4-amino-2-(4-(tert-butyl)phenyl)-6-(difluoromethyl)pyrimidine-5-carboxylic acid (170 mg, 0.53 mmol, crude yield 99%) as a yellow solid, which was used without further purification.

[0257] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)-6-(difluoromethyl)pyrimidine-5-carboxylic acid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.25(d,5.6 Hz,1H),8.85(d,J=8.1 Hz,1H),8.32(d,J=7.6 Hz,1H),8.21(d,J=8.6 Hz,2H),7.52(d,J=8.7 Hz,2H),7.19(dd,J=8.6,2.2 Hz,1H),7.01(d,J=8.6 Hz,1H),6.83(s,1H),6.73(s,1H),6.66-6.75(m,2H),6.26-6.33(m,2H),4.80(m,1H),4.73-4.60(m,2H),4.08-3.88(m,6H),3.28(d,J=15.1 Hz,1H),3.03-2.78(m,11H),2.35(s,3H),2.09-1.99(m,1H),1.97-1.86(m,1H),1.29(s,9H),1.17(d,J=6.7 Hz,3H).LCMS(Method 5-100 AB,6 min):RT =1.66 min,[M+H] + =979.3. Example 33 JPEG0007842812000079.jpg67107

[0258] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(2-fluoro-4-(tert-pentyl)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.49(s,1H),7.77-7.73(m,1H),7.29-7.27(m,1 H),7.20-7.05(m,3H),6.86-6.77(m,2H),6.61-6.48(m,2H),5.30-5.20(m,1H ),4.36-4.14(m,8H),3.63-3.60(m,1H),3.40-3.36(m,1H),3.21-3.01(m,9H) ,2.48(s,3H),1.74-1.71(m,2H),1.33(s,9H),0.74-0.70(m,3H).LCMS(Method 5-95 AB,ESI):R T =0.752 min,[M+H] + =1040.5. Example 34 JPEG0007842812000080.jpg67110

[0259] The compound was prepared as described in Example 54 by substituting 4-amino-2-(4-(cyclopropylmethyl)phenyl)-6-methylpyrimidine-5-carboxylic acid with 2-(3,3-dimethylbutoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine with 2-(4-(cyclopropylmethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan. The title compound was prepared as described in Example 1 by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(cyclopropylmethyl)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.18(d,J=7.6 Hz,2H),7.34(d,J=8.0 Hz,2H),7.12-7.04(m,1H),6.90-6.74(m,3H),6.59(s,1H),6.52-6.42(m,1H),5.22-5.13(m,1H),4.82-4.80( m,2H),4.27-3.91(m,6H),3.63-3.54(m,1H),3.25-3.14(m,3H),3.06(s,3H),3.03-2.95(m,2H),2.59(d,J=6.8 Hz,2H),2.47(s,3H),1.40-1.29(m.3H),1.07-0.95(s,1H),0.57-0.50(m,2H),0.26-0.19(m,2H)ppm.LCMS(Method 5-95 AB,ESI):R T =0.750 min,[M+H] + =1006.3. Example 35 JPEG0007842812000081.jpg69109

[0260] 2-amino-6-(4-(tert-butyl)-2-fluorophenyl)-4-methylnicotinic acid was produced using an approach similar to the procedure used to prepare 2-amino-6-(4-(tert-butyl)phenyl)-4-methylnicotinic acid as described in International Publication No. 2017084630 (the contents of which are incorporated herein by reference in their entirety). The title compound was prepared as described in Example 1 by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-amino-6-(4-(tert-butyl)-2-fluorophenyl)-4-methylnicotinic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.38(s,2H),7.70-7.68(m,1H),7.30-7.18(m,2H),7.03-6.55(m,6H),5.21(m, 1H),4.39-3.99(m,8H),3.62-3.38(m,3H),3.18-2.86(m,9H),2.32-2.28(m,3H),1.35(s,12H).LCMS(Method 5-95 AB,ESI):R T =0.783 min,[M+H] + =1025.5. Example 36 JPEG0007842812000082.jpg64107

[0261] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-butylphenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)8.45(s,1H),8.14(d,J=8.4 Hz 1H),7.28(d,J=8.4 Hz 1H),7.12-7.10(m,1H),6.91-6.86(m,2H),6.79(s,1H),6.59(s,1H),6.44(s,1H),5.19-5.16(m,1H),4.32-4.04(m,7H),3 .64-3.59(m,1H),3.23-3.07(m,10H),2.69-2.65(m,3H),2.47(s,3H),1.38-1.35(m,5H),0.97-0.93(m,3H).LCMS(Method 5-95 AB,ESI):R T =0.781min,[M+H] + =1008.5. Example 37 JPEG0007842812000083.jpg158164

[0262] The general procedure for the synthesis of methyl 6-(4-(tert-butyl)phenyl)-4-chloronicotinate was followed using the procedure described in J.Med.Chem.2013,56,1023-1040.

[0263] Step 1: 4,6-Dichloropyridine-3-carboxylate methyl (3072 mg, 14.91 mmol, 1.05 equivalent) and Pd(PPh3)4 (820 mg, 0.71 mmol, 0.05 mmol) were stirred in diglyme (13 mL) at room temperature for 15 minutes. Next, 4-tert-butylbenzeneboronic acid (2528 mg, 14.2 mmol, 1.0 equivalent) and a 2.0 M aqueous solution of K2CO3 (13.14 mL, 26.27 mmol, 1.85 equivalent) in IPA (15 mL) were added to this suspension. The mixture was then mixed at 95°C. oThe mixture was stirred in 1C for 2 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. A saturated aqueous solution of NaHCO3 and DCM were added to the crude reaction mixture. The phases were separated, the aqueous layer was extracted with additional DCM (2 × 40 mL), and the organic layers were combined. Next, the organic layers were washed with brine (2 × 30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Next, the solid was redissolved in DCM, silica gel (20 g) was added, and the resulting mixture was concentrated under reduced pressure to obtain a dried pack of yellow solid. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 0-30% ethyl phosphate in heptane) to obtain methyl 6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylate (2950 mg, 9.711 mmol, yield 68.4%) as a clear oil.

[0264] Step 2: 6-(4-tert--butylphenyl)-4-chloropyridine-3-carboxylate methyl (5000 mg, 16.46 mmol, 1.0 equivalent) was dissolved in DCM (54.8 mL), and 70% wt of 3-chloroperbenzoic acid (m-CPBA) (8521 mg, 24.69 mmol, 1.5 equivalents) was added. The reaction mixture was stirred at room temperature for 36 hours. The reaction mixture was diluted with a saturated solution of NaHCO3 and extracted with DCM (3 × 40 mL). The organic matter was combined, washed with water (1 × 40 mL) and brine (1 × 40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 10-80% ethyl phosphate in heptane) to obtain methyl 6-(4-tert-butylphenyl)-4-chloro-1-oxopyridine-3-carboxylate (3100 mg, 9.6942 mmol, yield 59%) as a beige solid.

[0265] Step 3: 6-(4-tert--butylphenyl)-4-chloro-1-oxidepyridine-1-ium-3-carboxylate methyl (3100 mg, 9.69 mmol, 1.0 equivalent) was dissolved in DMSO (19.4 mL), and sodium azide (1891 mg, 29.08 mmol, 3.0 equivalents) was added. Reaction mixture 50 oThe mixture was stirred in 1C for 30 minutes. The reaction mixture was cooled to room temperature, poured into water, and extracted with ethylacetate (3 × 40 mL). The organic matter was combined, washed with water (1 × 40 mL) and brine (1 × 40 mL), dried over anhydrous sodium 2SO4, filtered through a sintered funnel, and concentrated under reduced pressure to obtain methyl 4-azido-6-(4-tert-butylphenyl)-1-oxidepyridine-1-ium-3-carboxylate (2052 mg, 6.29 mmol, yield 64.9%) as a pale orange solid, which was carried over to the next step without purification.

[0266] The general procedure for the synthesis of methyl 6-(4-(tert-butyl)phenyl)-4-chloronicotinate was followed using the procedure described in Org. Lett. 2015, 17, 2948-2951.

[0267] Step 4: Add 4-azido-6-(4-tert-butylphenyl)-1-oxide-pyridine-1-ium-3-carboxylate methyl (500 mg, 1.53 mmol, 1.0 equivalent) and DCM (4.7 mL) to a flame-dried nitrogen-flush flask. Reaction mixture -70 o The mixture was cooled to C, and Et3N (0.43 mL, 3.06 mmol, 2.0 equivalents) was added, followed by oxalyl chloride (0.26 mL, 3.06 mmol, 2.0 equivalents). The mixture was then heated to -70°C. o The mixture was stirred in 1C for 3 hours. A saturated aqueous solution of NaHCO3 was added, and the reaction was diluted with toluene (30 mL). The phases were separated, and the aqueous layer was extracted with more toluene (2 × 10 mL). The organic layers were combined, washed with brine (2 × 20 mL), dried over Na2SO4, filtered through a sintered funnel, and concentrated under reduced pressure. The crude reaction mixture was then purified by flash chromatography (silica gel, 100-200 mesh, 0-50% toluene in heptane) to obtain methyl 4-azido-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylate (280 mg, 0.812 mmol, yield 53%) as a yellow solid.

[0268] Step 5: Triphenylphosphine (197.8 mg, 0.750 mmol, 1.0 equivalent) was added to a stirred solution of methyl 4-azido-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylate (260 mg, 0.750 mmol) in THF (7.54 mL). The reaction mixture was stirred at room temperature for 2 hours, then H₂O (2 mL) was added. The reaction mixture was stirred at room temperature for 12 hours. A saturated aqueous solution of NaHCO₃ and siRNA (20 mL) were added to separate the phases. The aqueous layer was extracted with more siRNA (2 × 10 mL). The organic layers were combined, washed with brine (2 × 20 mL), dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 0-100% ethyl ammonium in heptane) to obtain methyl 4-amino-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylate (180 mg, 0.565 mmol, yield 74.9%) as a white powder.

[0269] Step 6: 180 mg, 0.5600 mmol of methyl 4-amino-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylic acid was dissolved in 5.65 mL of THF, and 0.62 mL, 0.620 mmol, 1.1 equivalents of 1.0 M aqueous lithium hydroxide solution was added. The reaction mixture was stirred at room temperature for 24 hours. 40 mL of KHSO4 1N and 40 mL of siRNA were added, and the phases were separated. The aqueous layer was extracted with siRNA (2 × 40 mL). The organic layers were combined, washed with brine (3 × 40 mL), dried over Na2SO4, filtered through a sintered funnel, and the filtrate was concentrated under reduced pressure to isolate 4-amino-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylic acid (180 mg, 0.5906 mmol, yield 104%) as a white solid, which was carried over to the next step without purification.

[0270] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butylphenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylic acid. ¹H NMR (400 MHz, DMSO-d6+D2O) δ 8.31 (s, 1H), 7.79 (d, J=8.6 Hz, 2H), 7.47 (d, J=8.6 Hz, 2H), 7.08 (s, 1H), 7.09-7.04 (m, 1H), 6.89 (d, J=7.8 Hz,1H),6.74-6.64(m,2H),6.34(s,1H),6.24(s,1H),4.94-4.85(m,1H),4.60-4.58(m,1H),4.18-3.87(m,7) H),3.34-3.31(m,1H),3.27-3.19(m,1H),3.17-3.09(m,1H),3.05-2.76(m,8H),1.27(s,9H),1.16(d,J=7.0 Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.85 min,[M+H] + =1027.5. Example 38 JPEG0007842812000084.jpg67109

[0271] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(1,1-difluoro-2-methylpropyl)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)δ(ppm)8.42-8.34(m,3H),7.61-7.49(m,2H),7.21-7.03(m,1H),6.97-6.72(m,3H),6.68-6.56(m,1H),6.47(s,1H),5.20 -5.10(m,1H),4.84-4.73(m,2H),4.41-4.00(m,6H),3.75-3.32(m,3H), 3.27-2.91(m,8H),2.64-2.30(m,4H),1.45-1.25(m,3H),1.00(d,J=6.8 Hz,6H).LCMS(Method 5-95 AB,ESI):R T =0.645 min,[M+H] + =1045.0. Example 39 JPEG0007842812000085.jpg95164

[0272] THF (2.00 mL) and NH3 . To a solution of 2-(4-(tert-butyl)-2-formylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (140 mg, 0.30 mmol) in H2O (4.00 mL, 0.30 mmol), I2 (84.7 mg, 0.33 mmol) was added. The reaction mixture was 20 o The mixture was stirred in 1C for 2 hours. The solution was concentrated to dryness. The residue was partitioned between ethyl acetate (40 mL) and water (40 mL). The organic phase was washed with saturated Na2S2O3 solution (2 × 30 mL) and brine (30 mL). The organic phase was dried over Na2SO4 and concentrated to dryness. The residue was purified by preparative TLC (ethyl acetate:petroleum ether = 1:10) to obtain 2-(4-(tert-butyl)-2-cyanophenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (135 mg, yield 97.1%) as a yellow solid.

[0273] CAN (646 mg, 1.18 mmol) was added to a solution of 2-(4-(tert-butyl)-2-cyanophenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (135 mg, 0.29 mmol) in acetonitrile (6.0 mL) and water (3.0 mL). Reaction mixture 20 o The mixture was stirred in 1C for 30 minutes. The reaction mixture was partitioned between ethyl acetate (40.0 mL) and water (40.0 mL). The organic phase was washed with brine (2 × 40 mL), dried over Na₂SO₄, and concentrated to dryness. The residue was purified by preparative TLC (ethyl acetate:petroleum ether = 2:10) to obtain ethyl 4-amino-2-(4-(tert-butyl)-2-cyanophenyl)-6-methylpyrimidine-5-carboxylate (80.0 mg, yield 80.3%) as a yellow solid.

[0274] Step 3: Add LiOH to a solution of ethyl 4-amino-2-(4-(tert-butyl)-2-cyanophenyl)-6-methylpyrimidine-5-carboxylate (80.0 mg, 0.23 mmol) in THF (6.0 mL) and water (2.0 mL). . H2O (24.4 mg, 0.58 mmol) was added. The reaction mixture was 80 o The mixture was stirred in 1°C for 1 hour. The mixture was concentrated to remove methanol, and then water (20 mL) was added to the reaction mixture. The mixture was adjusted to pH=2 with 1 M HCl. The aqueous layer was extracted with ethyl acetate (2 × 20 mL). The combined organic layers were concentrated to dryness to obtain 4-amino-2-(4-(tert-butyl)-2-cyanophenyl)-6-methylpyrimidine-5-carboxylic acid (50 mg, yield 69.2%) as a white solid.

[0275] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-cyanophenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)8.53(s,1H),8.15-8.13(m,1H),7.85-7.75(m,2H),7.07(br,1H),7.00-6.84(m,2H),8.60(s,1H),8.48 (s,1H),5.20(s,1H),4.38-4.01(m,5H),3.63-3.60(m,1H),3.38-2.94(m,5H),2.50-2.45(m,3H),1.38(s,12H).LCMS(Method 5-100 AB,1.5 min):R T =0.633 min,[M+H] + =1033.7. Example 40 JPEG0007842812000086.jpg72110

[0276] Step 1: To a mixture of 1,4-dioxane (25 mL) and 4-tert-butylbenzeneboronic acid (500 mg, 2.81 mmol), 2-amino-4,6-dichloropyridine-3-carboxylate methyl (745 mg, 3.37 mmol), Pd(PPh3)4 (162 mg, 0.140 mmol), K3PO4 (894 mg, 4.21 mmol), and H2O (3 mL) were added. The reaction mixture was degassed with N2 gas and then stirred at 60°C for 18 hours. The reaction was quenched with saturated NaHCO3 aqueous solution (25 mL) and then extracted with ELISA (3 × 75 mL). The organic layers were combined, washed with brine, dried over Na2SO4, filtered through Celite, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-100% ethyl phosphate in heptane) to obtain methyl 2-amino-6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylate (565 mg, 1.77 mmol) as a yellow solid.

[0277] Step 2: A mixture of methyl 2-amino-6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylate (679 mg, 2.13 mmol) in 1,4-dioxane (5.6 mL) was mixed with 1 M aqueous LiOH (4.26 mL, 4.26 mmol). The reaction mixture was stirred at 60 °C for 15 hours and then cooled to room temperature. Diethyl ether was added, and the precipitate was collected by filtration to obtain 2-amino-6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylic acid (502 mg, yield 77.3%) as a pale yellow solid.

[0278] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-amino-6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylic acid. 1 H NMR(400MHz,DMSO+D2O)δ 8.35(s,2H),7.93(d,J=8.5Hz,2H),7.48(d,J=8.6Hz,2H),7.19(s,1H),7.07(d,J=9.7H) z,1H),6.89(d,J=8.6Hz,1H),6.75(s,1H),6.71(s,1H),6.37(s,1H),6.27(s,1H),4.94 - 4.87(m,1H),4.67 - 4.58(m,1H),4.17 - 4.10(m,1H),4.08 - 3.90(m,6H),3.41 - 3.09(m,4H),3.07 - 2.76(m,7H),1.29(s,9H),1.17(d,J=6.7Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.97 min,[M+H] + =1027.5. Example 41 JPEG0007842812000087.jpg69110

[0279] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-cyclobutylphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)8.48(s,1H),8.15(d,J=8.0 Hz 2H),7.35(d,J=8.0 Hz 2H),7.18-7.16(m,1H),6.97-6.95(m,2H),6.89(s,1H),6.82(s,1H),6.61(s, 1H),6.43(s,1H),5.23-5.19(m,1H),4.47-4.44(m,2H),4.33-4.08(m,6H),3.6 7-3.59(m,2H),3.42-3.36(m,1H),3.29-3.16(m,4H),3.09-3.04(m,5H),2.50( s,3H),2.40-2.38(m,2H),2.21-2.08(m,3H),1.94-1.89(m,1H),1.39(d,J=7.2 Hz,3H).LCMS(Method 5-95 AB,ESI):R T =0.737 min,[M+H] + =1006.6. Example 42 JPEG0007842812000088.jpg67107

[0280] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-6-methyl-2-(4-neopentylphenyl)pyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)δ(ppm)8.40(br s,1H),8.15(d,J=8.0 Hz,2H),7.24(d,J=8.0 Hz,2H),7.15-7.06(m,1H),6.85-6.78(m,3H),6.60(br,s,1H),6.48(s,1H),5.18(s,1H),4.50-4.22(m,6H),4.22- 4.01(m,6H),3.62-3.34(m,2H),3.24-2.99(m,8H),2.58-2.34(m,5H),1.38-1.35(m,3H),0.94(s,9H).LCMS(Method 5-95 AB,ESI):R T =0.798 min,[M+H] + =1022.3. Example 43 JPEG0007842812000089.jpg64107

[0281] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-hydroxyphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,DMSO)δ 8.87(d,J=8.1Hz,1H),8.33(d,J=7.8Hz,1H),8.17(d,J=8.4Hz,1H),7.21(d,J=8.7Hz,1H),7.02(d,J=8.6Hz,1H),6.97(dd,J=8.5,1.9 Hz,1H),6.87(d,J=1.9Hz,1H),6.75(s,1H),6.69(d,J=1.5Hz,1H),6.33(s,1H),6.30(s,1H),4.84(dd,J=9.6,4.1Hz,1H),4.74 - 4.63(m,2H),4.11 - 3.90(m,6H),3.34 - 3.25(m,1H),3.12 - 2.79(m,10H),2.37(s,3H),2.36(s,15H),2.10 - 1.89(m,2H),1.27(s,9H),1.18(d,J=6.7Hz,3H).LCMS(Method 5-100 AB,7 min):RT =1.64 min,[M+H] + =959.7. Example 44 JPEG0007842812000090.jpg69107

[0282] The compound was prepared as described in Example 54 by substituting 4-amino-2-(4-(tert-butyl)-2-hydroxyphenyl)-6-methylpyrimidine-5-carboxylic acid with 2-(3,3-dimethylbutoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine with 2-(2-(benzyloxy)-4-(tert-butyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan. The title compound was prepared as described in Example 1 by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-hydroxyphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.45(s,1H),8.29-8.19(m,1H),7.11-7.00(m,1 H),6.99-6.71(m,5H),6.66-6.56(m,1H),6.50(s,1H),5.23-5.09(m,1H),4.83 -4.52(m,1H),4.41(s,1H),4.31-3.94(m,6H),3.66-3.54(m,1H),3.42-3.34( m,1H),3.29-2.93(m,9H),2.51-2.36(m,3H),1.40-1.27(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.791 min,[M+H] + =1024.9. Example 45 JPEG0007842812000091.jpg64104

[0283] Step 1: To a stirred ice-cold DCM (50 mL) solution of 4-(1,1-dimethylpropyl)phenol (1.00 g, 6.09 mmol, 1.0 equivalent) under nitrogen, pyridine (981 uL, 12.18 mmol, 2.0 equivalents) was added, followed by the dropwise addition of trifluoromethanesulfonic anhydride (1.23 mL, 7.31 mmol, 1.2 equivalents). The mixture was stirred at 0°C for 30 minutes, then warmed to room temperature. The reaction product was washed with 1.0 M aqueous KHSO4 (30 mL), then saturated aqueous NaHCO3 (2 × 30 mL), and saturated brine aqueous solution (30 mL). The solution was dried over anhydrous MgSO4, filtered, and the filtrate was concentrated under reduced pressure. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 0-100% ethyl phosphate in heptane) to obtain [4-(1,1-dimethylpropyl)phenyl]trifluoromethanesulfonate (1800 mg, 6.075 mmol, 99.7% yield) as a clear oil.

[0284] Step 2: To a stirred solution of [4-(1,1-dimethylpropyl)phenyl]trifluoromethanesulfonate (400 mg, 1.35 mmol, 1.0 equivalent) in anhydrous DMSO (6 mL), bis(pinacolate)diborone (377 mg, 1.48 mmol, 1.1 equivalent), followed by KOAc (132 mg, 1.35 mmol, 1.0 equivalent). The mixture was stirred and sprayed with argon for 1 hour. Next, PdCl2(dppf)·CH2Cl2 (49 mg, 0.067 mmol, 0.05 equivalent) was added, and argon spraying was continued for 15 minutes. The mixture was kept under argon and heated to 80°C for 3 hours. The reaction mixture was cooled to room temperature, diluted with siRNA (75 mL), and washed with water (50 mL) and saturated brine aqueous solution (4 × 25 mL). The organic layer was dried over MgSO4, filtered, and the filtrate was concentrated under reduced pressure. The crude reaction mixture was then purified by flash chromatography (silica gel, 100-200 mesh, 0-100% ethyl phosphate in heptane) to obtain 2-[4-(1,1-dimethylpropyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (310 mg, 1.13 mmol, yield 83.7%) as a white solid.

[0285] Step 3: In a flame-dried nitrogen-flushed vial, 2-[4-(1,1-dimethylpropyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (336 mg, 1.23 mmol, 1.3 equivalents), K2CO3 (261 mg, 1.88 mmol, 2.0 equivalents), 4-amino-2-chloro-6-methylpyrimidine-5-carboxylate methyl (190 mg, 0.9400 mmol, 1.0 equivalent), and PdCl2·(dppf)CH2Cl2 (77 mg, 0.0900 mmol, 0.1 equivalents) were added. The solid was suspended in a mixture of 1,4-dioxane (2.36 mL) and water (0.24 mL) and degassed with nitrogen for 5 minutes. The vial was sealed and 100°C. o The reaction mixture was heated in C for 2 hours. The reaction mixture was cooled to room temperature, and saturated aqueous solution of NaHCO3 and ethylacetal were added. The layers were separated, and the aqueous layer was extracted with ethylacetal (2 × 10 mL). The organic layers were combined, washed with brine (2 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. Next, the crude reaction mixture was purified by flash chromatography (silica gel, 100-200 mesh, 0-60% ethylacetal in heptane) to obtain methyl 4-amino-2-[4-(1,1-dimethylpropyl)phenyl]-6-methylpyrimidine-5-carboxylate (80 mg, 0.255 mmol, yield 27%) as a white solid.

[0286] Step 4: In a scintillation vial equipped with a magnetic stirring rod, methyl 4-amino-6-chloro-2-[4-(1,1-dimethylpropyl)phenyl]pyrimidine-5-carboxylate (70 mg, 0.21 mmol, 1.0 equivalent), THF (2.1 mL), and 1.0 M aqueous lithium hydroxide solution (0.23 mL, 0.2300 mmol, 1.1 equivalents) were added. The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain lithium 4-amino-6-chloro-2-[4-(1,1-dimethylpropyl)phenyl]pyrimidine-5-carboxylate (67 mg, 0.2095 mmol, 99.9% yield) as a bright yellow solid, which was used directly without purification.

[0287] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with lithium 4-amino-6-chloro-2-[4-(1,1-dimethylpropyl)phenyl]pyrimidine-5-carboxylate. 1 H NMR (400 MHz, DMSO-d6 + D2O) δ 8.11 (d, J = 7.9 Hz, 2H), 7.51 (d, J = 8.0 Hz, 2H), 7.19 (d, J = 10.0 Hz, 1H), 6.99 (d, J = 8.8 Hz, 1H), 6.70 (s, 2H), 6.28 (s, 1H), 6.20 (s, 1H), 5.00 - 4.88 (m, 1H), 4.73 - 4.55 (m, 2H), 4.11 - 3.90 (m, 6H), 3.37 - 3.23 (m, 2H), 3.16 - 2.90 (m, 4H), 2.91 - 2.75 (m, 4H), 2.67 - 2.62 (m, 1H), 2.42 (s, 3H), 2.36 (s, 12H), 1.63 (q, J = 7.9 Hz, 2H), 1.25 (s, 6H), 1.16 (d, J = 6.7 Hz, 3H), 0.60 (t, J = 7.4 Hz, 3H). LCMS (Method 5-100 AB, 7 min): R T = 1.83 min, [M+H] + = 1022.7. Example 46 JPEG0007842812000092.jpg72112

[0288] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)pyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)8.81-8.72(m,1H),8.47(s,1H),8.17(d,J=6.8 Hz,2H),7.47(d,J=6.8 Hz,2H),7.06(s,1H),6.78-6.67(m,2H),6.48-6.40(m,2H),4.77-3.48(m,12H),3.13-2.69(m,6H),1.49-1.22(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.778 min,[M+H] + =994.4. Example 47 JPEG0007842812000093.jpg65107

[0289] Compound 20 was prepared from the title compound using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(1,1-dimethylindan-5-yl)-6-methylpyrimidine-5-carboxylic acid. 1 1H NMR (400MHz, DMSO+D2O)δ 8.32(s,3H),8.07-8.05(m,2H),7.22(d,J=8.8Hz,1H),7.08(d,J=9.6Hz,1H) ,6.89(d,J=8.5Hz,1H),6.72(s,1H),6.69(s,1H),6.37(s,1H),6.24(s,1H), 4.85-4.79(m,1H),4.62-4.55(m,1H),4.11-4.06(m,1H),4.06-3.96(m,4H), 3.95-3.91(m,2H),3.26-3.19(m,1H),3.02-2.80(m,12H),2.32(s,3H),2.09 - 1.83(m,4H),1.20(s,6H),1.16(d,J=7.0Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.59 min,[M+H] + =962.5. Example 48 JPEG0007842812000094.jpg116164

[0290] A mixture of 4-(tert-butyl)-2-methylphenol (400 mg, 2.44 mmol) in dichloromethane (5.0 mL) contains 0 o Pyridine (0.39 mL, 4.87 mmol) and Tf2O (0.61 mL, 3.65 mmol) were added to C. Reaction mixture 0 o The mixture was stirred in 1C for 2 hours. The reaction mixture was diluted with dichloromethane (30 mL). The organic layer was washed with 1 M HCl (10 mL), saturated NaHCO3 (20 mL), and brine (2 × 50 mL). The organic layer was dried over Na2SO4 and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 4-(tert-butyl)-2-methylphenyltrifluoromethanesulfonate (710 mg, yield 98.4%) as a colorless oil.

[0291] A mixture of 4-(tert-butyl)-2-methylphenyltrifluoromethanesulfonate (710.0 mg, 2.4 mmol), potassium acetate (705 mg, 7.19 mmol), Pd(dppf)Cl2 (178 mg, 0.24 mmol), and bis(pinacolate)diborone (791 mg, 3.11 mmol) in DMF (10.0 mL) was subjected to nitrogen for 80°C. o The mixture was stirred in C for 16 hours. The mixture was diluted with ethyl acetate (80 mL), filtered, and the filtrate was washed with brine (3 × 80 mL). The organic layer was concentrated under vacuum. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(4-(tert-butyl)-2-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (530 mg, yield 80.7%) as a white solid.

[0292] A mixture of 2-(4-(tert-butyl)-2-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaneborolane (530 mg, 1.93 mmol), 2-chloro-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (681 mg, 2.03 mmol), Pd(dppf)Cl2 (70.7 mg, 0.10 mmol), and Na2CO3 (615 mg, 5.80 mmol) is suspended in 1,4-dioxane (6.0 mL) and water (0.60 mL), purged with N2 (15 psi), and 100°C. o The reaction mixture was heated in C for 16 hours. The reaction mixture was diluted with ethyl acetate (20 mL). The reaction mixture was washed with brine (2 × 20 mL), dried over Na₂SO₄, and concentrated. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(4-(tert-butyl)-2-methylphenyl)-4-((4-methoxybenzyl))amino)-6-methylpyrimidine-5-carboxylate ethyl (500 mg, yield 57.8%) as a colorless oil.

[0293] Step 4: Dissolve 2-(4-(tert-butyl)-2-methylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (200 mg, 0.45 mmol), (diacetoxyiodo)benzene (216 mg, 0.67 mmol), Cu(OTFA)2 (12.9 mg, 0.04 mmol), and Pd(OAc)2 (5.02 mg, 0.02 mmol) in acetic acid (0.30 mL) and acetic anhydride (4.50 mL) in air at 80°C. oThe mixture was stirred in 14°C for 24 hours. The solvent was removed under vacuum. The residue was diluted with saturated NaHCO3 solution (15.0 mL) and extracted with ethyl acetate (3 × 30 mL). The organic layers were combined, washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum) to obtain 2-(2-acetoxy-4-(tert-butyl)-6-methylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (110 mg, yield 48.7%) as a pale yellow oil.

[0294] Step 5: A solution of 2-(2-acetoxy-4-(tert-butyl)-6-methylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (110 mg, 0.22 mmol) in trifluoroacetic acid (3.0 mL, 0.22 mmol) is prepared in 75 ml. o The mixture was stirred in C for 16 hours. The mixture was concentrated to dryness and purified by column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum ether) to obtain 4-amino-2-(4-(tert-butyl)-2-hydroxy-6-methylphenyl)-6-methylpyrimidine-5-carboxylate ethyl (74.0 mg, yield 99%) as a white solid.

[0295] Step 6: Add NaOH (34.5 mg, 0.86 mmol) to a methanol (5.00 mL) solution of ethyl 4-amino-2-(4-(tert-butyl)-2-hydroxy-6-methylphenyl)-6-methylpyrimidine-5-carboxylate (74.0 mg, 0.22 mmol) in water (1.00 mL). The reaction mixture was 80 o The mixture was stirred in 1°C for 1 hour. The mixture was concentrated to dryness and diluted with water (30 mL). The mixture was adjusted to pH=5 with 1 M HCl, and the aqueous layer was extracted with ethyl acetate (2 × 60 mL). The combined organic layers were dried over Na₂SO₄ and concentrated under vacuum to obtain 4-amino-2-(4-(tert-butyl)-2-hydroxy-6-methylphenyl)-6-methylpyrimidine-5-carboxylic acid (65.0 mg, yield 95.7%) as a white solid.

[0296] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-hydroxy-6-methylphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)8.46(s,1H),7.10-7.00(m,1H),6.87-6.76(m,5H),6.61(s,1H),6.47(s,1H),5.19-5. 16(m,1H),4.37-3.90(m,8H),3.63-3.58(m,3H),3.19-2.99(m,10H),2.51(s,3H),2.46(s,3H),1.35(d,J=6.8 Hz,3H),1.30(s,9H).LCMS(Method 5-95 AB,ESI):R T =0.670 min,[M+H] + =1009.6. Example 49 JPEG0007842812000095.jpg69107

[0297] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-amino-6-(4-(tert-butyl)phenyl)-4-methylnicotinic acid. 1H NMR(400MHz,DMSO+D2O)δ 8.93(d,J=9.3Hz,1H),8.37(d,J=9.2Hz,1H),7.81(d,J=8.5Hz,2H),7.60(d,J=8.5Hz,2H),7.24 - 7.15(m,2H),7.02(d,J=8.9Hz,1H),6.76 - 6.65(m,2H),6.31(s,1H),6.23(s,1H),5.02 - 4.92(m,1H),4.77 - 4.60(m,2H),4.14 - 3.86(m,6H),3.42 - 3.23(m,2H),3.18 - 2.76(m,9H),2.37(s,12H),1.31(s,9H),1.17(d,J=6.7Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.77 min,[M+H] + =1007.5. General Procedure C: JPEG0007842812000096.jpg115164

[0298] Step 1: To a solution of compound 5 (2.0 g, 2.5 mmol) (synthesized according to general procedure A) in DMF (12 mL), tert-butyl(5S)-5-[(3-nitrophenyl)sulfonyloxymethyl]-2-oxo-oxazolidine-3-carboxylate (1.36 g, 3.38 mmol) (synthesized according to general procedure D, starting with (S)-3-amino-1,2-propanediol) and potassium carbonate (1.04 g, 7.52 mmol) were added under an inert atmosphere at 25°C. The reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was added dropwise to water / saturated aqueous solution NaHCO3 (1:1) with stirring. An orange solid precipitated and was recovered by filtering. The solid was purified by column chromatography (silica gel, 100-200 mesh, 25-100% ethyl acetate in a (1:1) DCM / heptane mixture) to obtain compound 6 (1.99 g, yield 79.6%) as a white solid.

[0299] Step 2: Compound 6 (1.99 g, 2 mmol) was dissolved in methanol (12.7 mL), the reaction flask was purged with nitrogen, and then palladium catalyst (159 mg, 0.15 mmol) with 10% wt carbon was added. The reaction mixture was purged with hydrogen and incubated under 1 atmosphere of hydrogen for 25 minutes. o The mixture was stirred in 1C for 3 hours. Before adding Cbz-O-succinimide (0.5 g, 2 mmol), the reaction flask was purged into the solution by nitrogen bubbling for 15 minutes. The reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was filtered through Celite and concentrated under reduced pressure. The residue was diluted with ethyl acetate (100 mL), washed with water (2 × 100 mL) and brine (2 × 100 mL), dried over Na₂SO₄, and concentrated to dryness to obtain compound 7 (1.81 g, yield 99.8%) as a grayish-white solid.

[0300] Step 4: p-toluenesulfonic acid (29 mg, 0.155 mmol) was added to a solution of compound 8 (1.72 g, 1.55 mmol) in methanol (17 mL). Reaction mixture 25 o The mixture was stirred in 16°C for 16 hours. The reaction mixture was added dropwise to water / saturated aqueous NaHCO3 (1:1) with stirring. An orange solid precipitated and was recovered by filtration. The solid was dissolved in DCM, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain crude compound 9 (1.52 g, 100% yield) as a grayish-white solid.

[0301] Step 5: Solution of compound 9 (1.52 g, 1.55 mmol) in DMF (15 mL) and methanol (0.565 mL) under an inert atmosphere (N2) at -15 o The solution was cooled to C. Cesium carbonate (1.07 g, 3.28 mmol) was added to the cold solution, and the reaction mixture was adjusted to -20 to -15°C. o The mixture was stirred in C for 6 hours. The filtrate was concentrated to obtain crude compound 6 (35.0 g, yield 97.4%) as a white solid. Reaction mixture -15 oThe solution was diluted with HCl (20 mL) and water (20 mL) in 14C. The phases were separated, and the organic phase was washed with water (2 × 30 mL) and brine (2 × 30 mL), dried on Na₂SO₄, and concentrated under reduced pressure. The solid was purified by column chromatography (silica gel, 100-200 mesh, 2-10% MeOH in DCM) to obtain compound 10 (1.07 g, 75% yield) as a white solid. General Procedure D: JPEG0007842812000097.jpg68164

[0302] Step 1: (R)-3-amino-1,2-propanediol (107 mL, 1.38 mol) was placed in a 5 L flask, and both 10 volumes of THF (1260 mL) and water (1260 mL) were added. The mixture was stirred at room temperature for 20 minutes until the reactants were completely dissolved. Next, the solution was cooled to 0°C, and potassium carbonate (229.4 g, 1.66 mol) was added. Finally, benzyl chloroformate (237 mL, 1.66 mol) was added dropwise to the reaction mixture using an addition funnel, while maintaining the internal temperature below 8°C for 60 minutes. After the reaction was complete (2 hours), the layers were separated. The organic layer was concentrated under reduced pressure (to half) while the aqueous phase was extracted three times with ELISA (3 × 1260 mL). All organic matter was combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to 2-3 volumes. Heptane (1260 mL) was added to the residue and stirred at 10-20°C for 2-3 hours. The resulting white solid was filtered and washed with heptane. The solid was transferred to a flask and dried under high vacuum to obtain compound 2 (325 g, yield 104%) as a white solid.

[0303] Compound 2 (175 g, 779 mmol) was placed in a 5 L round-bottom flask with a three-neck thermometer and dissolved in THF (2631 mL). The solution was cooled to 0°C, and potassium tert-butoxide (96.13 g, 857 mmol) was added gradually while maintaining the temperature between 0 and 10°C. Once the addition of the reagents was complete, reaction mixture 25 oThe mixture was heated to 1°C and stirred for 3 hours. Once complete, the reaction mixture was cooled again using an ice bath, and while maintaining the internal temperature at approximately 10°C, 4M HCl in the dioxane was added to the reaction mixture until the pH reached 5-6. Next, reaction mixture 25 o The mixture was stirred in C for 30 minutes. The precipitate was filtered and washed twice with MeCN (700 mL). The wet cake was placed in a 2 L Erlenmeyer flask and stirred with 1.5 L of MeCN for 30 minutes. The solid was filtered again and washed twice with MeCN (700 mL). All organic compounds were combined and concentrated to 2-3 V under reduced pressure below 45°C. MTBE (1.5 L) was added to the resulting suspension, and the suspension was concentrated again under reduced pressure below 45°C until it reached 2-3 V. 1-1.5 L of MTBE was added to the suspension and stirred at room temperature for 30 minutes. The solid was then collected by filtering and washed with MTBE (1 L). The solid was dried under vacuum to obtain compound 3 (85.5 g, yield 93.8%) as a grayish-white solid.

[0304] Compound 3 (75 g, 640 mmol) was placed in a 2 L round-bottom flask, and 6 V MeCN (450 mL), followed by pyridine (155 mL, 1.92 mol). The resulting solution was cooled to 0°C. Next, while maintaining the temperature below 10°C, 3-nitrobenzenesulfonyl chloride (149 g, 673 mmol) was added little by little. The reaction mixture was stirred at 0°C for 2 hours. After completion, the reaction mixture was concentrated to 2-3 V under reduced pressure. Next, MTBE (10 V, 750 mL) was added to the flask, and the resulting mixture was concentrated to 2-3 V under reduced pressure. 1.5 L of saturated NaHCO3 solution (20 V) was added. The mixture was vigorously stirred at 10-20°C for 30 minutes. The resulting mixture was filtered and washed twice with water (1000 mL x 2). The moist cake was collected and returned to the round-bottom flask. 1.5 L of saturated NaHCO3 solution (20 V) was added. The mixture was vigorously stirred at 10-20°C for 30 minutes. The resulting mixture was filtered and washed twice with water (1000 mL x 2). The moist cake was collected and returned to a round-bottom flask. Water (10V, 750 mL) and MTBE (5V, 375 mL) were added, and the resulting two-phase suspension was vigorously stirred for 30 minutes. The solid was filtered and washed twice with MTBE (400 mL x 2). The cake was dried under high vacuum to obtain compound 4 (178 g, 92% yield) as a grayish-white solid.

[0305] Compound 4 (182.7 g, 604 mmol) was added to a 5 L round-bottom flask together with 4-dimethylaminopyridine (7.38 g, 60.4 mmol), and MeCN (1.1 L) was added. Reaction mixture 0 o Cool to C and set the reaction temperature to 0 o While maintaining the temperature below C, tert-butyl dicarbonate (237.5 g, 1.1 mol) was added. Reaction mixture 0 oThe mixture was stirred in C for 1 hour. Once complete, the reaction mixture was concentrated under reduced pressure. 500 mL of MeOH was added to the mixture and concentrated under reduced pressure to form a dark orange gum. 700 mL of MeOH was added to the mixture. The reaction flask was placed under sonication for 3 minutes. A white precipitate formed, which was stirred at room temperature for 30 minutes. The solid was collected by filtration and washed with cold MeOH to obtain compound 5 (191.4 g, yield 78.7%) as a white solid. 98.95%ee.1H NMR(400 MHz,CDCl3)δ 8.77(t,J=1.9 Hz,1H),8.56(ddd,J=8.3,2.2,1.0 Hz,1H),8.25(ddd,J=7.9,1.7,1.1 Hz,1H),7.85(t,J=8.0 Hz,1H),4.75 - 4.67(m,1H),4.36(dd,J=11.5,3.5 Hz,1H),4.30(dd,J=11.5,4.2 Hz,1H),4.05(dd,J=10.6,9.2 Hz,1H),3.82(dd,J=10.6,6.2 Hz,1H),1.53(s,9H). Example 50 JPEG0007842812000098.jpg67105

[0306] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)-6-methylpyrimidine-5-carboxylic acid, and replacing compound 14 with compound 10 as described in procedure C. 1H NMR(400 MHz,DMSO-d6+D2O)δ(ppm)δ 9.28(d,J=3.9 Hz,1H),8.91(d,J=6.7 Hz,1H),8.31(d,J=5.0 Hz,1H),8.06(d,J=7.9 Hz,2H),7.63(d,J=8.2 Hz,2H),7.19(d,J=7.8 Hz,1H),7.00(d,J=8.7 Hz,1H),6.69(d,J=6.2 Hz,2H),6.28(s,1H),6.17(s,1H),5.06 - 4.93(m,1H),4.73 - 4.59(m,2H),4.17 - 3.94(m,8H),3.41 - 3.33(m,1H),3.32 - 3.22(m,1H),3.17 - 3.06(m,1H),3.05 - 2.95(m,2H),2.92 - 2.76(m,1H),2.85(s,3H),2.42(s,3H),2.39(s,9H),1.29(s,9H),1.16(d,J=6.4 Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.62 min,[M+H] + =1008.5. Example 51 JPEG0007842812000099.jpg65105

[0307] The title compound was prepared from compound 20 using the procedure of Example 1, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-6-(4-tert-butylphenyl)-2-chloropyridine-3-carboxylic acid prepared as described in Example 37. 1H NMR(400 MHz,DMSO-d6+D2O)δ 8.34(s,3H),7.80(d,J=8.5 Hz,2H),7.48(d,J=8.6 Hz,2H),7.09(s,1H),7.06(d,J=8.8 Hz,1H),6.87(d,J=8.6 Hz,1H),6.74(s,1H),6.67(s,1H),6.37(s,1H),6.24(s,1H),4.86-4.80(m,1H),4.65 - 4.55(m,1H),4.13 - 3.94(m,7H),3.26-3.21(m,1H),3.09 - 2.76(m,10H),2.03-1.92(m,2H),1.28(s,9H),1.15(d,J=6.7 Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.59 min,[M+H] + =962.5. Example 52 JPEG0007842812000100.jpg110164

[0308] Step 1: Add TiCl4 (14.3 mL, 127 mmol) to a DCM (45.0 mL) in a three-necked RB flask and heat under a nitrogen atmosphere at -78°C. o The temperature was maintained at C. Then, while maintaining the temperature, 127 mL (127 mmol) of 1 M Zn(CH3)2 in toluene was added. The resulting orange-brown solution was then heated to -78°C. o The mixture was vigorously stirred in 1°C for 1 hour. A solution of 5-bromo-2,3-dihydro-1H-inden-1-one (4.50 g, 21.3 mmol) in DCM (45.0 mL) was added dropwise to the above mixture. The reaction solution was heated to -78°C. o Stir in C for 2 hours, then -10 oThe mixture was heated to 16°C and stirred for 16 hours. An ice-cold saturated NH4Cl solution was added dropwise to quench the reaction mixture. The organic matter was separated, the aqueous layer was extracted with ethyl acetate (3 × 100 mL), and the organic layer was washed with brine (3 × 100 mL). The organic matter was combined, dried over Na2SO4, and then concentrated to dryness. Next, the crude product was purified by flash column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 5-bromo-1,1-dimethyl-2,3-dihydro-1H-indene (4.80 g, yield 99.8%) as a yellow oil.

[0309] Step 2: Mixture of 5-bromo-1,1-dimethyl-2,3-dihydro-1H-indene (5.7g, 25.3mmol), Pd(dppf)Cl2 (926mg, 1.27mmol), KOAc (7.45g, 75.9mmol), and pinacol diboronate (9.64g, 37.9mmol) in DMF (57.0mL) under N2 at 80°C. o The mixture was stirred in 1C for 3 hours. The solvent was removed, and the residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.5 g, yield 94.3%) as a white solid.

[0310] Step 3: Purge a mixture of 2-(1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaneborolane (1.95 g, 7.15 mmol), 2-chloro-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl 2-chloro-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate (2.00 g, 5.96 mmol), Pd(dppf)Cl2 (0.22 g, 0.30 mmol), and Na2CO3 (1.89 g, 17.8 mmol) with N2 (15 psi) and 100 oThe mixture was heated at 16°C for 16 hours. After filtering, 50.0 mL of ethyl acetate was added to the mixture. The mixture was washed with brine (2 × 50.0 mL). The organic matter was dried over Na₂SO₄ and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(6-acetoxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (2.50 g, yield 94.2%) as a colorless oil.

[0311] Step 4: The solutions of 2-(6-acetoxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (1.00 g, 2.24 mmol), PhI(OAc)2 (1.08 g, 3.37 mmol), Cu(OTFA)2 (64.9 mg, 0.22 mmol), and Pd(OAc)2 (25.1 mg, 0.11 mmol) in HOAc (1.00 mL) and Ac2O (15 mL) are heated to 80°C in the air. oThe mixture was stirred in 1C for 16 hours. The solvent was removed under vacuum. The residue was diluted with saturated NaHCO3 solution (30.0 mL), extracted with ethyl acetate (3 × 100 mL), washed with brine (100 mL), dried over Na2SO4, and concentrated under vacuum. The given crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 5-(5-(ethoxycarbonyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-2-yl)-1,1-dimethyl-2,3-dihydro-1H-indene-4,6-diyldiacetate (300 mg, 0.5957 mmol, yield 26.5%), 2-(6-acetoxy-1,1-di Methyl-2,3-dihydro-1H-inden-5-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (300 mg, 0.5957 mmol, yield 26.5%) and 2-(4-acetoxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (330 mg) were obtained as a pale yellow oily substance.

[0312] Step 5: Dissolve 5-(5-(ethoxycarbonyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-2-yl)-1,1-dimethyl-2,3-dihydro-1H-indene-4,6-diyldiaacetate (330 mg, 0.590 mmol) in TFA (5.00 mL) for 75 minutes. o The mixture was stirred in 16°C for 16 hours. The mixture was concentrated. Ethyl acetate (40 mL) was added. The organic layer was washed with NaHCO3 (aqueous solution, 30.0 mL) and brine (30.0 mL), dried over Na2SO4, and concentrated. The crude substance was purified by column chromatography on silica gel (16% ethyl acetate in petroleum ether) to obtain 4-amino-2-(4,6-dihydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylate ethyl (160 mg, 76.2% yield) as a yellow solid.

[0313] Step 6: To a solution of ethyl 4-amino-2-(4,6-dihydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylate (160 mg, 0.450 mmol) in MeOH (5.00 mL) and water (1.00 mL), NaOH (71.6 mg, 1.79 mmol) was added. The reaction mixture was 80 o The mixture was stirred in 1C for 3 hours. The pH of the mixture was adjusted to 2 with 1M HCl (aqueous solution). The mixture was partitioned between ethyl acetate (50.0 mL) and water (30.0 mL). The organic layer was washed with brine (40.0 mL), dried over Na2SO4, and filtered. The filtrate was concentrated to obtain 4-amino-2-(4,6-dihydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylic acid (100 mg, yield 67.8%) as a yellow solid.

[0314] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4,6-dihydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.42(br s,1H),7.25-6.95(m,1H),6.91-6.45(m,5H),6.21(s,1H),5.26-5.08(m,1H),4.64-3.88(m,8H),3.6 8-3.34(m,2H),3.26-2.93(m,9H),2.79-2.70(m,2H),2.43(s,3H),1.95-1.83(m,2H),1.36(d,J=6.4 Hz,3H),1.23(s,6H).LCMS(Method 5-95AB,ESI):R T =0.799 min,[M+H] + =1052.5. Example 53 JPEG0007842812000101.jpg69107

[0315] Step 1: Sodium methoxide (25 wt.%, 16.4 mL, 71.5 mmol in methanol) was added to a solution of 4-tert-butylbenzamidine (4.20 g, 23.8 mmol) in methanol (11.9 mL). The reaction mixture was stirred at room temperature for 10 minutes, then diethyl malonate (3.62 mL, 23.8 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours, then concentrated under reduced pressure. H2O was added, followed by concentrated HCl to obtain an acidic pH. The resulting mixture was extracted with ELISA (3×). The organic layers were combined, washed with water and brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-(4-tert-butylphenyl)pyrimidine-4,6-diol (4.08 g, yield 70.1%) as a grayish-white solid. The crude material was used in the next step without further purification.

[0316] Step 2: DMF (1.33 mL, 17.2 mmol) was added to POCl3 (21.5 mL, 230 mmol) at 0°C. The reaction mixture was stirred at 0°C for 1 hour. Next, this solution was added to 2-(4-tert-butylphenyl)pyrimidine-4,6-diol (4.01 g, 16.4 mmol), and the reaction mixture was stirred at room temperature for 30 minutes, then at 100°C for 16 hours. Next, the mixture was cooled to room temperature and poured into ice / water. The resulting mixture was extracted with SiO(3×). The organic layers were combined, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% siRNA in heptane) to obtain 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carbaldehyde (2.94 g, yield 57.9%) as a grayish-white solid.

[0317] Step 3: A solution of sodium chlorite (1.20 g, 13.3 mmol) in water (4.2 mL) was added to tert-butanol (21 mL) and a solution of 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carbaldehyde (2.94 g, 9.51 mL) and sulfamic acid (1.29 g, 13.3 mmol) in water (8.4 mL). The reaction mixture was stirred at room temperature for 5 hours, then the additional portions of sulfamic acid (260 mg, 2.68 mmol) and sodium chlorite (240 mg, 2.65 mmol) were added. The reaction mixture was stirred at room temperature for a further 2 hours. Water was added, and the mixture was extracted with ELISA (3×). The organic layers were combined, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carboxylic acid (3.09 g, 99.9% yield) as a grayish-white solid. The crude material was used in the next step without further purification.

[0318] Step 4: Potassium carbonate (3.28 g, 23.8 mmol) was added to a solution of 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carboxylic acid (3.09 g, 9.50 mmol) in DMF (47.5 mL). The reaction mixture was stirred at room temperature for 15 minutes, then ethane iodide (1.91 mL, 23.8 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours, then diluted with ethyl acetate. The resulting mixture was washed with water (2x) and brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude substance was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in heptane) to obtain 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carboxylic acid ethyl acetate (2.95 g, yield 87.9%) as a grayish-white solid.

[0319] Step 5: A solution of 2-(4-tert-butylphenyl)-4,6-dichloropyrimidine-5-carboxylate ethyl (800 mg, 2.26 mmol) and 2M ammonia in iPrOH (24.0 mL, 48.0 mmol) was stirred at room temperature for 16 hours. Next, H2O was added and the mixture was extracted with ELISA (3x). The organic layers were combined, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 4-amino-2-(4-tert-butylphenyl)-6-chloropyrimidine-5-carboxylate ethyl (765 mg, quantitatively) as a colorless oil. The crude material was used in the next step without further purification.

[0320] Step 6: A 1N lithium hydroxide aqueous solution (2.40 mL, 2.40 mmol) was added to a solution of ethyl 4-amino-2-(4-tert-butylphenyl)-6-chloropyrimidine-5-carboxylic acid (200 mg, 0.599 mmol) in THF (6.0 mL). The reaction mixture was stirred at 50°C for 16 hours and then cooled to room temperature. A 1N HCl aqueous solution was added, and the resulting mixture was extracted with a 4:1 solution of CHCl3 / iPrOH (3×). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 190 mg of 4-amino-2-(4-tert-butylphenyl)-6-chloropyrimidine-5-carboxylic acid as a grayish-white solid.

[0321] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butylphenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-tert-butylphenyl)-6-chloropyrimidine-5-carboxylic acid. 1H NMR(400MHz,DMSO+D2O)δ 8.34(s,1H),8.16(d,J=8.5Hz,2H),7.52(d,J=8.6Hz,2H),7.07(d,J=8.9Hz,1H),6.89(d, J=8.4Hz,1H),6.72(s,1H),6.70(s,1H),6.35(s,1H),6.25(s,1H),4.94-4.88(m,1H),4.66 - 4.56(m,1H),4.19 - 4.11(m,1H),4.08 - 3.99(m,3H),3.99 - 3.93(m,3H),3.40 - 3.29(m,1H),3.25 - 3.10(m,2H),3.07 - 2.98(m,2H),2.97 - 2.79(m,6H),1.28(s,9H),1.17(d,J=6.8Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.88 min,[M+H] + =1028.2. Example 54 JPEG0007842812000102.jpg110164

[0322] Step 1: To a solution of 5-bromopyridine-2-ol (2.00 g, 11.49 mmol) in DMF (10.0 mL), 1-bromo-3,3-dimethylbutane (2.85 g, 17.2 mmol) and Ag2CO3 (9.51 g, 34.5 mmol) were added. The reaction mixture was stirred at 110°C for 2 hours. The reaction mixture was diluted with ethyl acetate (40 mL). After filtration, the filtrate was washed with brine (3 × 40 mL) and dried on Na2SO4. After filtration, the filtrate was concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum) to obtain 5-bromo-2-(3,3-dimethylbutoxy)pyridine (1.00 g, yield 33.7%) as a yellow oil.

[0323] Step 2: A mixture of 5-bromo-2-(3,3-dimethylbutoxy)pyridine (1.20 g, 4.65 mmol), KOAc (1.37 g, 14.0 mmol), Pd(dppf)Cl2 (345 mg, 0.46 mmol), and bis(pinacolate)diborone (1.77 g, 6.97 mmol) in DMF (10.0 mL) was stirred under nitrogen at 80°C for 16 hours. The reaction mixture was diluted with ethyl acetate (20 mL), then filtered, and the filtrate was washed with brine (3 × 20 mL). The organic layer was concentrated to dryness under vacuum. The residue was purified by column chromatography (silica gel, 100-200 mesh, 50-100% ethyl acetate in petroleum) to obtain 2-(3,3-dimethylbutoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (480 mg, yield 33.8%) as a yellow oily substance.

[0324] Step 3: A mixture of 2-(3,3-dimethylbutoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (300 mg, 0.98 mmol), 2-chloro-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (347 mg, 1.03 mmol), Pd(dppf)Cl2 (36.0 mg, 0.05 mmol), and Na2CO3 (312 mg, 2.95 mmol) in a mixture of 1,4-dioxane (6.0 mL) and water (0.60 mL) was purged with N2 (15 psi) and heated at 100°C for 16 hours. After filtration, 20 mL of ethyl acetate was added to the mixture. The mixture was washed with brine (2 × 20 mL). The organic layer was dried over Na2SO4 and concentrated to dryness. The crude substance was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(6-(3,3-dimethylbutoxy)pyridine-3-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (370 mg, yield 78.7%) as a yellow solid.

[0325] Step 4: To a solution of ethyl 2-(6-(3,3-dimethylbutoxy)pyridine-3-yl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate (175.0 mg, 0.37 mmol) in acetonitrile (4.00 mL) and water (2.00 mL), cerium ammonium nitrate (802 mg, 1.46 mmol) was added. The reaction mixture was stirred at 20°C for 30 minutes. The reaction mixture was partitioned between ethyl acetate (20 mL) and water (20 mL). The organic layer was washed with brine (2 × 20 mL), dried over Na₂SO₄, and concentrated to dryness. The crude product was purified by preparative TLC (ethyl acetate:petroleum ether = 1:10, Rf = 0.3) to obtain ethyl 4-amino-2-(6-(3,3-dimethylbutoxy)pyridine-3-yl)-6-methylpyrimidine-5-carboxylate (100 mg, yield 76.3%) as a yellow solid.

[0326] Step 5: NaOH (44.6 mg, 1.12 mmol) was added to a solution of ethyl 4-amino-2-(6-(3,3-dimethylbutoxy)pyridine-3-yl)-6-methylpyrimidine-5-carboxylic acid (100 mg, 0.28 mmol) in methanol (5.00 mL) and water (1.00 mL). The reaction mixture was stirred at 80°C for 1 hour. The mixture was concentrated to dryness. The pH of the mixture was adjusted to 2 with HCl (1 M). The aqueous layer was extracted with ethyl acetate (2 × 60 mL), and the combined organic layers were concentrated to obtain 4-amino-2-(6-(3,3-dimethylbutoxy)pyridine-3-yl)-6-methylpyrimidine-5-carboxylic acid (80 mg, yield 86.8%) as a white solid.

[0327] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(6-(3,3-dimethylbutoxy)pyridin-3-yl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)9.04(s,1H),8.51-8.42(m,2H),7.12-6.99(m,1H),6.89-6.69(m,3H),6.55(s, 1H),5.27-5.10(m,1H),4.83-4.73(m,2H),4.53-4.29(m,3H),4.18-4.07(m,3H),4.05-3.92(m,1H),3 .67-3.52(m,1H),3.44-3.37(m,1H),3.36-3.33(m,1H),3.29-3.19(m,2H),3.18-3.09(m,2H),3.07(s ,3H),3.00-2.91(m,1H),2.45(s,3H),1.79-1.67(m,2H),1.41-1.28(m,2H),1.00(s,9H).LCMS(Method 10-80AB,ESI):R T =1.868 min,[M+H] + =1054.6. Example 55 JPEG0007842812000103.jpg65107

[0328] The title compound was prepared using the procedure of Example 1, by replacing compound 14 with compound 20 and 1-(4-(tert-butylphenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-tert-butylphenyl)-6-chloropyrimidine-5-carboxylic acid prepared as described in Example 53. 1H NMR(400MHz,DMSO+D2O)δ 8.87(d,J=8.0Hz,1H),8.30(d,J=7.7Hz,1H),8.15(d,J=8.1 Hz,2H),7.52(d,J=8.2Hz,2H),7.19(d,J=8.5Hz,1H),7.00(d,J=8.6Hz,1H),6.71(s,2H),6.30(s,1H),6.25(s,1H),4.86 - 4.80(m,1H),4.73 - 4.58(m,2H),4.20 - 3.96(m,5H),3.85 - 3.64(m,3H),3.35 - 3.23(m,1H),3.15 - 2.77(m,11H),2.74 - 2.55(m,1H),2.46 - 2.36(m,2H),2.40(s,12H),2.38 - 2.20(m,1H),2.13 - 1.86(m,2H),1.28(s,9H),1.18(d,J=7.1Hz,3H)LCMS(Method 5-100 AB,7 min):R T =1.63 min,[M+H] + =963.6 Example 56 JPEG0007842812000104.jpg69107

[0329] 60 ml of ethanol containing 2-(4-(tert-butyl)phenyl)-4-chloro-6-methylpyrimidine-5-carboxylate ethyl (150 mg, 0.45 mmol), TEA (188 μL, 1.35 mmol), and methylamine (33.4 mg, 0.50 mmol). o The mixture was stirred in C for 1 hour. The reaction mixture was diluted with ethyl acetate (40 mL). The organic layer was washed with brine (2 × 30 mL), dried on Na2SO4, and concentrated to dryness. The residue was separated into TLC (10% ethyl acetate in petroleum ether, R f Purified by (0.4), 4-amino-2-(4-(tert-butyl)-2-methylphenyl)-6-methylpyrimidine-5-carboxylate ethyl (125 mg, yield 85.0%) was obtained as a white solid.

[0330] Step 5: 4-amino-2-(4-(tert-butyl)-2-methylphenyl)-6-methylpyrimidine-5-carboxylate ethyl (120 mg, 0.37 mmol) was dissolved in MeOH (10.0 mL) and water (2.0 mL). NaOH (58.6 mg, 1.47 mmol) was added, and the mixture was heated to 80°C. o The mixture was stirred in 1C for 2 hours. The reaction mixture was concentrated and the pH was adjusted to pH=3 with 1M HCl. The reaction mixture was partitioned between ethyl acetate (50.0 mL) and water (50.0 mL). The aqueous layer was then extracted with ethyl acetate (50.0 mL x 2). The combined organic layers were dried over Na2SO4 and concentrated to obtain 2-(4-(tert-butyl)phenyl)-4-methyl-6-(methylamino)pyrimidine-5-carboxylic acid (100 mg, yield 91%) as a white solid.

[0331] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)-4-methyl-6-(methylamino)pyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)8.38(s,1H),8.24(d,J=7.6 Hz,2H),7.50(d,J=8.4Hz,2H),7.10-7.04(m,1H),6.50-6.77(m,2H),6.59(s,1H),6.48(s,1H),5.20-5.16(m,1H),4.37 -3.97(m,8H),3.61-3.58(m,1H),3.38-3.35(m,1H),3.19-2.98(m,12H),2.44(s,3H),1.36-1.34(m,12H). 5-95 AB,ESI):R T =0.794 min,[M+H] + =1022.5. Example 57 JPEG0007842812000105.jpg69107

[0332] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(6-hydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylic acid, which was prepared using the procedure described in Example 52. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.47(br s,1H),8.11(s,1H),7.02(br s,1H),6.88-6.70(m,3H),6.65(s,1H),6.59(br s,1H),6.52(br s,1H),5.18(br d,J=8 Hz 1H),4.44-3.95(m,8H),3.6(br d,J=10.4 Hz,1H),3.43-3.32(m,1H),3.26-2.96(m,9H),2.86-2.79(m,2H),2.44(s,3H),1.96-1.88(m,2H),1.35(br d,J=6.4 Hz,3H),1.25(br s,6H).LCMS(Method 5-95 AB,ESI):R T =0.783 min,[M+H] + =1036.5. Example 58 JPEG0007842812000106.jpg120164

[0333] Step 1: Add 0 to the mixture of 5-(tert-butyl)-2-hydroxybenzaldehyde (4.30 g, 24.1 mmol) in DCM (60 mL). o At C, pyridine (3.89 mL, 48.3 mmol) and trifluoromethanesulfonic anhydride (6.09 mL, 36.2 mmol) were added. Reaction mixture 0 oThe mixture was stirred in 1C for 2 hours. The reaction mixture was diluted with ethyl acetate (100 mL). The organic layer was successively washed with 1 M HCl (50 mL), saturated NaHCO3 (50 mL), and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum) to obtain 4-(tert-butyl)-2-formylphenyltrifluoromethanesulfonate (4.50 g, yield 60.1%) as a yellow oil.

[0334] Step 2: Dissolve the mixture of 4-(tert-butyl)-2-formylphenyltrifluoromethanesulfonate (3.90 g, 12.6 mmol), bis(pinacolate)diborone (4.78 g, 18.85 mmol), potassium acetate (3.75 g, 37.7 mmol), and Pd(PPh3)Cl2 (882 mg, 1.26 mmol) in DMF (40.0 mL) for 80 minutes. o The mixture was stirred in 1C under N2 (15 psi) for 16 hours. The mixture was filtered and diluted with ethyl acetate (80 mL). The organic layer was washed with brine (2 × 100 mL), dried over Na2SO4, and concentrated to dryness. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain the crude product. The crude product was purified by preparative HPLC (using a gradient of acetonitrile and water (containing 0.225% formic acid)) to obtain 5-(tert-butyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (1.30 g, yield 35.9%) as a yellow solid.

[0335] Step 3: Purge a mixture of 5-(tert-butyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaneborolan-2-yl)benzaldehyde (300 mg, 1.04 mmol), ethyl 2-chloro-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (402 mg, 1.2 mmol), Pd(dppf)Cl2 (76.2 mg, 0.10 mmol), and Na2CO3 (331 mg, 3.12 mmol) in a mixed solvent of 1,4-dioxane (10.0 mL) and water (1.0 mL) with N2 (15.0 psi), and 100 o The mixture was heated at 16°C for 16 hours. After filtering, 20 mL of ethyl acetate was added to the reaction mixture. The mixture was washed with brine (2 × 30 mL), dried over Na₂SO₄, and concentrated to dryness. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-5% ethyl acetate in petroleum) to obtain 2-(4-(tert-butyl)-2-formylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (220 mg, yield 45.8%) as a white solid.

[0336] Step 4: To a solution of 2-(4-(tert-butyl)-2-formylphenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (220 mg, 0.48 mmol) in methanol (10.0 mL), 10% Pd / C (101 mg, 0.10 mmol) was added. The mixture was heated under hydrogen (50 psi) for 40°C. o The mixture was stirred in C for 6 hours. The mixture was filtered, and the filtrate was concentrated to obtain 2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (180 mg, yield 81.5%) as a white solid.

[0337] Step 5: To a solution of 2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-4-((4-methoxybenzyl)amino)-6-methylpyrimidine-5-carboxylate ethyl (180 mg, 0.39 mmol) in acetonitrile (8.0 mL) and water (4.0 mL), cerium ammonium nitrate (851 mg, 1.55 mmol) was added. The reaction mixture was then mixed for 20 minutes. o The mixture was stirred in C for 30 minutes. The reaction mixture was partitioned between ethyl acetate (40 mL) and water (40 mL). The organic phase was washed with brine (2 × 40 mL), dried over Na₂SO₄, and concentrated to dryness. The crude product was separated into TLC (ethyl acetate:petroleum ether = 1:10, TLC: 15% ethyl acetate in petroleum, R₂). f Purified by (0.3), 4-amino-2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-6-methylpyrimidine-5-carboxylate ethyl (100 mg, yield 75%) was obtained as a yellow solid.

[0338] Step 6: NaOH (46.59 mg, 1.16 mmol) was added to a solution of ethyl 4-amino-2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-6-methylpyrimidine-5-carboxylate (100 mg, 0.29 mmol) in methanol (10.0 mL) and water (3.0 mL). The reaction mixture was 80 o The mixture was stirred in 1°C for 1 hour. The mixture was concentrated to remove methanol. Water (20 mL) was added to the mixture, and the pH of the mixture was adjusted to 2 with 1 M HCl. The aqueous layer was extracted with ethyl acetate (2 × 20 mL). The combined organic layers were concentrated to obtain 4-amino-2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-6-methylpyrimidine-5-carboxylic acid (80.0 mg, yield 87.1%) as a white solid.

[0339] Step 7: The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-(hydroxymethyl)phenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)8.42(s,2H),7.90(d,J=8.4 Hz,2H),7.55-7.48(m,1H),7.46(br,1H),7.09-7.82(m,3H),6.63(s,1H),6.50(s,1H),5.22-5.19(m,1H),4.96- 4.26(m,1H),4.25(br,1H),4.24-4.15(m,4H),3.22-3.05(m,4H),2.45-2.44(m,3H),1.39(s,12H).LCMS(Method 5-95 AB,ESI):R T =0.713 min,[M+H] + =1038.5. Example 59 JPEG0007842812000107.jpg69104

[0340] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-methylphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)8.41(s,1H),7.49-7.39(m,1H),7.36-7.26(m,2H), 7.18-7.01(m,1H),6.94-6.73(m,3H),6.67-6.57(m,1H),6.49-6.28(m,1H) ,5.23-5.10(m,1H),4.74-4.50(m,2H),4.45-3.91(m,6H),3.66-3.31(m,3 H),3.26-2.70(m,8H),2.52-2.29(m,6H),1.42-1.25(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.627 min,[M+H] + =1022.8. Example 60 JPEG0007842812000108.jpg69107

[0341] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-3-fluorophenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.42(br s,1H),7.46-7.36(m,1H),7.18-7.00(m,1H),6.95-6.72(m,3H),6.59(s,1H),6.46(s,1H),5.23-5.08(m,1H),4.80- 4.77(m,2H),4.50-3.94(m,6H),3.68-3.33(m,3H),3.29-2.95(m,8H),2.47(s,3H),1.45-1.31(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.774 min,[M+H] + =1026.6. Example 61 JPEG0007842812000109.jpg69107

[0342] Step 1: Add DIPEA (16.4 mL, 94.17 mmol, 2.0 equivalents) to a flame-dried nitrogen-flash flask, followed by chloromethyl methyl ether (MOMCl) (5.4 mL, 70.6 mmol, 1.5 equivalents) to a solution of 5-tert-butyl-2-iodophenol (13.0 g, 47.08 mmol, 1.0 equivalent) in DCM (157 mL) at room temperature. Stir the reaction mixture at room temperature for 16 hours. Add saturated aqueous NaHCO3 to the reaction mixture (40 mL) at 0°C. Extract the mixture with DCM (3 × 40 mL). Combine the organic layers, dry over Na2SO4, filter through a sintered funnel, and concentrate the filtrate under reduced pressure. The crude substance was purified by filtering it on a silica gel (800g) pad, which eluted 20% phenylethylamine from heptane, to obtain 4-tert-butyl-1-iodo-2-(methoxymethoxy)benzene (13.8g, 43.1 mmol, yield 91.5%) as a yellow oily substance.

[0343] In a flame-dried nitrogen-flash flask, 4-tert-butyl-1-iodo-2-(methoxymethoxy)benzene (13.0 g, 40.6 mmol, 1.0 equivalent), 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (28.9 mL, 142.11 mmol, 3.5 equivalents), and anhydrous THF (203 mL) were added. The solution was then prepared at -78°C. o The mixture was cooled to 13C, and a 2.5 M solution of n-BuLi in heptane (48.7 mL, 121.8 mmol, 3.0 equivalents) was added dropwise. The reaction mixture was stirred for 3 hours. A saturated aqueous solution of NaHCO3 was added at -78°C, the flask was warmed to room temperature, and the mixture was diluted with siRNA (300 mL). The phases were separated, and the aqueous layer was extracted with more siRNA (2 × 100 mL). The organic layer was then washed with brine (2 × 200 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica gel, 100-200 mesh, 0-30% siRNA in heptane) to obtain 2-[4-tert-butyl-2-(methoxymethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3542 mg, 11.061 mmol, yield 27%) as a white solid.

[0344] The title compound was prepared using the procedure of Example 17, by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid and 4-tert-butylbenzeneboronic acid with 2-[4-tert-butyl-2-(methoxymethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, as described in Example 37, with 4-amino-6-((4-tert-butyl)-2-hydroxyphenyl)-2-chloropyridine-3-carboxylic acid. 1H NMR(400 MHz,DMSO-d6+D2O)δ 8 8.32(s,2H),7.65(d,J=8.7 Hz,1H),7.21(s,1H),7.07(d,J=8.0 Hz,1H),6.95(dd,J=8.4,1.9 Hz,1H),6.89(d,J=8.3 Hz,1H),6.85(d,J=1.9 Hz,1H),6.73(s,1H),6.70(s,1H),6.36(s,1H),6.25(s,1H),4.92-4.88(m,1H),4.64 - 4.53(m,1H),4.11(dd,J=5.6,4.7 Hz,1H),4.05-3.91(m,6H),3.37-3.30(m,1H),3.26-3.18(m,1H),3.19 - 3.09(m,1H),3.02-2.88(m,6H),2.86-2.75(m,2H),1.24(s,9H),1.16(d,J=6.8 Hz,3H).LCMS(Method 5-100 AB,7 min):R T =2.03 min,[M+H] + =1043.5. Example 62 JPEG0007842812000110.jpg69107

[0345] The compound was prepared as described in Example 54 by substituting 4-amino-2-(4-(tert-butyl)-2-fluorophenyl)-6-methylpyrimidine-5-carboxylic acid with 2-(3,3-dimethylbutoxy)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine with 2-(4-(tert-butyl)-2-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan. The title compound was prepared as described in Example 1 by substituting 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-2-fluorophenyl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400MHz,MeOH-d4)δ(ppm)7.75-7.69(m,1H),7.34-7.26(m,1H),7.25-7.17(m,1 H),7.12-7.01(m,1H),6.96-6.73(m,3H),6.63-6.54(m,1H),6.44(s,1H),5.21-5.10 (m,1H),4.82-4.65(m,4H),4.44(s,1H),4.32-3.87(m,6H),3.66-3.48(m,1H),3.42- 3.32(m,1H),3.25-2.87(m,8H),2.49-2.32(m,3H),1.39-1.29(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.769 min,[M+H] + =1026.5. Example 63 JPEG0007842812000111.jpg67105

[0346] The general procedure for the synthesis of ethyl 6-(4-(tert-butyl)phenyl)-4-chloro-2-methylnicotinate was followed using the procedure described in J.Med.Chem.2013,56,1023-1040.

[0347] Step 1: 4,6-Dichloro-2-methylpyridine-3-carboxylate ethyl (966 mg, 4.13 mmol, 1.05 equivalent) and Pd(PPh3)4 (227 mg, 0.2000 mmol, 0.05 equivalent) were stirred in Diglyme (3.6 mL) at room temperature for 15 minutes. Next, 4-tert-butylbenzeneboronic acid (700 mg, 3.93 mmol, 1.0 equivalent) in IPA (4.2 mL) was added to this suspension, followed by a 2 M aqueous solution of K2CO3 (3.64 mL, 7.27 mmol, 1.85 equivalent). The mixture was then mixed in 95 ml. oThe mixture was stirred in 1C for 2 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. A saturated aqueous solution of NaHCO3 and DCM were added to the reaction mixture, and the layers were separated. The aqueous layer was extracted with DCM (2 × 40 mL), and the organic layer was combined with it. Next, the organic layer was washed with brine (2 × 30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain ethyl 6-(4-tert-butylphenyl)-4-chloro-2-methylpyridine-3-carboxylate (1.02 g, 3.0738 mmol, yield 78.2%) as a yellow solid, which was then removed without purification and proceeded to the next step.

[0348] Step 2: Palladium acetate (35.5 mg, 0.1600 mmol, 0.15 equivalents) was added to a flame-dried nitrogen-flushed vial along with ethyl 6-(4-tert-butylphenyl)-4-chloro-2-methylpyridine-3-carboxylate (350 mg, 1.05 mmol, 1.0 equivalent) and Catacxium A (113.5 mg, 0.3200 mmol, 0.3 equivalents). The powder was dissolved in ethylene glycol (2.64 mL) and 1,4-dioxane (2.64 mL) (1:1), and the solution was sprayed with a nitrogen stream. Potassium (tert-butoxycarbonylamino)methyl-trifluoroboranoid (1000 mg, 4.22 mmol, 4.0 equivalents) and DIPEA (735 μL, 4.22 mmol, 4.0 equivalents) were added, and the suspension was heated in an oil bath for 100°C. o The mixture was heated to 14°C. After 4 hours, the reaction mixture was cooled to room temperature, and saturated aqueous solution of NaHCO3 and ethyl phosphate were added. The phases were separated, the aqueous layer was extracted with more ethyl phosphate (2 × 10 mL), and the organic layers were combined. Next, the organic layers were washed with brine (2 × 20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain a black residue. The crude material was purified by flash chromatography (silica gel, 100-200 mesh, 0-40% ethyl phosphate in heptane) to obtain 4-[(tert-butoxycarbonylamino)methyl]-6-(4-tert-butylphenyl)-2-methylpyridine-3-carboxylate ethyl phosphate (175 mg, 0.3399 mmol, yield 32%) as a dark red oil.

[0349] Step 3: 4-[(tert-butoxycarbonylamino)methyl]-6-(4-tert-butylphenyl)-2-methylpyridine-3-carboxylate ethyl (145 mg, 0.3400 mmol, 1.0 equivalent) was added to a scintillation vial equipped with a magnetic stirring rod. This was dissolved in THF (1.7 mL). Next, a 1.0 M aqueous solution of lithium hydroxide (0.37 mL, 0.37 mmol, 1.1 equivalents) was added. The reaction mixture was then heated to 50°C for 1 hour. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain 4-[(tert-butoxycarbonylamino)methyl]-6-(4-tert-butylphenyl)-2-methylpyridine-3-carboxylate lithium (135 mg, 0.3388 mmol, yield 99.7%) as a yellow foam / solid. This was used directly in the next step.

[0350] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-[(tert-butoxycarbonylamino)methyl]-6-(4-tert-butylphenyl)-2-methylpyridine-3-carboxylate lithium. 1 H NMR(400 MHz,DMSO-d6+D2O)δ 7.94(d,J=8.4 Hz,2H),7.84(s,1H),7.52(d,J=8.5 Hz,2H),7.18(d,J=8.9 Hz,1H),6.99(d,J=8.7 Hz,1H),6.72(s,1H),6.67(s,1H),6.28(s,1H),6.14(s,1H),5.09-5.03(m,1H),4.71-4.58(m,2H),4.18-3.91(m,9H),3 .90-3.75(m,2H),3.41-3.22(m,1H),3.12-2.95(m,3H),2.85-2.77(m,4H),2.54(s,3H),2.39(s,12H),1.27(s,9H),1.21 - 1.11(m,3H).LCMS(Method 5-100 AB,7 min):R T =1.48 min,[M+H] + =1021.6. Example 64 JPEG0007842812000112.jpg69107

[0351] The title compound was prepared as described in Example 50 by replacing tert-butyl(5R)-5-[(3-nitrophenyl)sulfonyloxymethyl]-2-oxo-oxazolidine-3-carboxylate in step 3 with tert-butyl(5S)-5-[(3-nitrophenyl)sulfonyloxymethyl]-2-oxo-oxazolidine-3-carboxylate. 1 H NMR(400MHz,DMSO+D2O)δ 8.98 - 8.90(m,1H),8.40 - 8.31(m,1H),8.12(d,J=8.5Hz,2H),7.62(d,J=8.4Hz,2H),7.21(d,J=7.3Hz,1H),7.04(d,J=8.6Hz,1H),6.76 - 6.67(m,2H),6.30(s,1H),6.21(s,1H),5.00-4.96(m,1H),4.74 - 4.62(m,2H),4.14 - 3.95(m,5H),3.93 - 3.82(m,1H),3.40 - 3.24(m,2H),3.18 - 2.93(m,4H),2.92 - 2.83(m,4H),2.82 - 2.75(m,1H),2.46(s,3H),2.37(s,9H),1.31(s,9H),1.22 - 1.15(m,3H).LCMS(Method 5-100 AB,7 min):R T =1.67 min,[M+H] + =1008.6. Example 65 JPEG0007842812000113.jpg69105

[0352] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-(aminomethyl)-6-(4-(tert-butyl)-2-hydroxyphenyl)-2-methylnicotinic acid. The compound was prepared as described in Example 63 by replacing 4-(aminomethyl)-6-(4-(tert-butyl)-2-hydroxyphenyl)-2-methylnicotinic acid and 4-tert-butylbenzeneboronic acid with 2-[4-tert-butyl-2-(methoxymethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane. 1 H NMR(400MHz,DMSO+D2O)δ 8.32(s,2H),8.07(s,1H),7.93(d,J=8.5Hz,1H),7.08(d,J=8.8Hz,1H),6.99(d,J=8.4Hz,1H),6.94 - 6.87(m,2H),6.74(s,1H),6.71(s,1H),6.37(s,1H),6.25(s,1H),5.08(dd,J=8.8,4.6Hz,1H),4.66 - 4.58(m,1H),4.21 - 4.12(m,1H),4.09 - 3.88(m,6H),3.85(s,2H),3.39 - 3.29(m,1H),3.27 - 3.19(m,1H),3.17 - 3.08(m,1H),3.06 - 2.76(m,8H),2.54(s,3H),1.27(s,9H),1.22 - 1.15(m,3H).LCMS(Method 5-100 AB,7 min):R T =1.68 min,[M+H] + =1037.6. Example 66 JPEG0007842812000114.jpg69107

[0353] The general procedure for the synthesis of 5-bromo-1,1-dimethyl-2,3-dihydro-1H-indene was followed using the procedure described in Chem. Ber. 1985, 118, 1050.

[0354] Step 1: Titanium(IV) chloride (1.71 mL, 15.64 mmol, 2.2 equivalents) and DCM (14.2 mL) were added to a flame-dried nitrogen-flush flask. The solution was cooled to -30°C and stirred for 5 minutes. Next, 15.64 mL, 15.64 mmol, 2.2 equivalents of 1 M dimethylzinc in toluene was added dropwise, and the reaction mixture was stirred for 30 minutes. Next, 5-bromo-1-indanone (1500 mg, 7.11 mmol, 1.0 equivalent) was added all at once to a slurry that had been stirred overnight at -30°C to room temperature. Next, the reaction mixture was cooled to 0°C, ice cubes were added little by little, followed by NaHCO3. The reaction mixture was diluted with ethyl acetate, the solid was decanted, and the layers were separated. The aqueous layer was extracted with ethyl acetate (3 × 40 mL). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was diluted with DCM and filtered through a silica gel pad (200 g). The silica gel was washed off with 50% siRNA in hexane. The solution was evaporated to dryness to obtain crude 5-bromo-1,1-dimethylindan (1495 mg, 6.6409 mmol, yield 93.4%) as a translucent oil, which was then removed without further purification and proceeded to the next step.

[0355] Step 2: 5-bromo-1,1-dimethylindan (1500 mg, 6.66 mmol, 1.0 equivalent) and bis(pinacolate)diborone (3384 mg, 13.33 mmol, 2.0 equivalents) were added to a flame-dried nitrogen-flushed vial. The solid was dissolved in anhydrous toluene (22 mL), and nitrogen was sprayed over the solution for 15 minutes. KOAc (2.59 g, 19.99 mmol, 3.0 equivalents) was added, followed by PdCl2(dppf)·CH2Cl2 (272 mg, 0.330 mmol, 0.05 equivalents). The vial was sealed with a microwave cap, and the solution was then heated to 100°C and stirred overnight. The reaction mixture was cooled to room temperature and diluted with  (100 mL). The organic layer was washed with saturated aqueous NaHCO3 solution, then dried over brine and Na2SO4, and filtered through a sintering funnel. Silica gel was added, and the suspension was evaporated under reduced pressure to obtain a black solid. The solution was then purified by flash chromatography (silica gel, 100-200 mesh, 0-20% ethyl ammonium in heptane) to obtain 2-(1,1-dimethylindan-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1324 mg, 4.8642 mmol, yield 73%) as a sticky orange solid.

[0356] Step 3: In a flame-dried sealed tube, methyl 4-amino-2-chloro-6-methylpyrimidine-5-carboxylate (150 mg, 0.7400 mmol, 1.0 equivalent), 1,4-dioxane (3.7201 mL), and water (0.3618 mL) were added. Nitrogen was passed through the solution for 10 minutes. Next, 2-(1,1-dimethylindan-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (243 mg, 0.8900 mmol, 1.2 equivalents), PdCl2(dppf)·CH2Cl2 (60.8 mg, 0.0700 mmol, 0.1 equivalent), and K2CO3 (206 mg, 1.49 mmol, 2.0 equivalents) were added to the reaction mixture. The tube was sealed with a microwave cap and heated in an oil bath at 95°C. oThe mixture was heated in 1C for 12 hours. After this period, the solution was cooled to room temperature, and saturated aqueous solution of NaHCO3 and ethylacetal were added. The phases were separated, the aqueous layer was extracted with ethylacetal (2 × 10 mL), and the organic layers were combined. Next, the organic layers were washed with brine (2 × 20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude reaction mixture was then purified by flash chromatography (silica gel, 100-200 mesh, 0-50% ethylacetal in heptane) to obtain methyl 4-amino-2-(1,1-dimethylindan-5-yl)-6-methylpyrimidine-5-carboxylate (81 mg, 0.2601 mmol, yield 35%) isolated as a yellow solid.

[0357] Step 4: 180 mg, 0.5800 mmol of methyl 4-amino-2-(1,1-dimethylindan-5-yl)-6-methylpyrimidine-5-carboxylate was dissolved in THF (1.93 mL), and a 1.0 M aqueous solution of lithium hydroxide (2.31 mL, 2.31 mmol, 4.0 equivalents) was added. Next, the reaction mixture was 50 o The mixture was heated in C and stirred overnight. The reaction mixture was cooled, and 1.0 M KHSO4 (40 mL) and siRNA (40 mL) were added. The phases were separated, and the aqueous layer was extracted with siRNA (2 × 40 mL). The organic layers were combined, washed with brine (3 × 40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 4-amino-2-(1,1-dimethylindan-5-yl)-6-methylpyrimidine-5-carboxylic acid (165 mg, 0.5549 mmol, 96% yield) as a white solid, which was then carried directly to the next step without purification.

[0358] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(1,1-dimethylindan-5-yl)-6-methylpyrimidine-5-carboxylic acid. 1H NMR(400 MHz,DMSO-d6+D2O)δ 8.36(s,1H),8.10-8.08(m,2H),7.26-7.14(m,1H),7.03(d,J=8.8 Hz,1H),6.85(d,J=8.2 Hz,1H),6.73(s,1H),6.69(s,1H),6.35(s,1H),6.24(s,1H),4.98-4.85(m,1H),4.69-4.50(m,1H),4.21-4.11( m,1H),4.07-3.87(m,6H),3.38-3.29(m,1H),3.24-3.08(m,2H),3.04-2.79(m,8H),2.34(s,3H),1.88(t,J=7.1 Hz,2H),1.27-1.11(m,9H).LCMS(Method 5-100 AB,7 min):R T =1.66 min,[M+H] + =1020.5. Example 67 JPEG0007842812000115.jpg67105

[0359] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)-3-fluoro-2-hydroxyphenyl)-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.42(br s,2H),8.00(d,J=7.6 Hz,1H),7.12-6.51(m,7H),5.26-5.16(m,1H),4.83-4.80(m,1H),4.49-3.94(m,7H), 3.69-3.32(m,3H),3.25-2.92(m,8H),2.42(s,3H),1.43-1.33(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.792 min,[M+H] + =1042.6. Example 68 JPEG0007842812000116.jpg69107

[0360] Step 1: To a mixture of 1,4-dioxane (58 mL) and 4-t-butylbenzeneboronic acid (1150 mg, 6.46 mmol, 1 equivalent), 2-amino-4,6-dichloropyridine-3-carboxylate methyl (1571 mg, 7.11 mmol, 1.1 equivalent), Pd(PPh3)4 (373 mg, 0.32 mmol, 0.05 equivalent), K3PO4 (2057 mg, 9.69 mmol, 1.5 equivalent), and H2O (5 mL) were added, and the mixture was degassed with N2 gas. 60 o After stirring in 18 hours, the reaction was quenched with a saturated aqueous solution of NaHCO3 (25 mL), diluted, and extracted with toluene (3 × 75 mL). The organic layer was washed with brine, dried over Na2SO4, filtered through Celite, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 10-50% toluene in heptane) to obtain methyl 2-amino-4,6-dichloropyridine-3-carboxylate (1510 mg, 4.74 mmol, yield 73%) as a yellow solid.

[0361] Step 2: N2 degassing 1,4-dioxane (7.8419 mL) contains a mixture of 2-amino-6-(4-tert-butylphenyl)-4-chloropyridine-3-carboxylate methyl (500 mg, 1.57 mmol, 1 equivalent), methylboronic acid (282 mg, 4.71 mmol, 3 equivalents), 1,1'-bis(diphenylphosphino)ferrocenedichloropalladium(II) (115 mg, 0.16 mmol, 0.1 equivalent), and K3PO4 (999 mg, 4.71 mmol, 3 equivalents). o The mixture was stirred in 1°C for 1 hour. The reaction product was quenched with a saturated aqueous solution of NaHCO3 (10 mL), diluted, and then extracted with toluene (3 × 50 mL). The organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 10-50% toluene in heptane) to obtain methyl 2-amino-6-(4-tert-butylphenyl)-4-methylpyridine-3-carboxylate (342 mg, 1.15 mmol, yield 73%) as a yellow solid.

[0362] Step 3: N-iodosuccinimide (396 mg, 1.76 mmol, 1.5 equivalents) was added to a solution of 2-amino-6-(4-tert-butylphenyl)-4-methylpyridine-3-carboxylate methyl (350 mg, 1.17 mmol, 1 equivalent) in DMF (12 mL). Reaction mixture 65 o The mixture was stirred in 1C for 30 hours. The reaction product was quenched with a saturated aqueous solution of NaHCO3 (20 mL), diluted, and then extracted with siRNA (3 × 50 mL). The organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-80% siRNA in heptane) to obtain methyl 2-amino-6-(4-tert-butylphenyl)-5-iodo-4-methylpyridine-3-carboxylate (279 mg, 0.66 mmol, yield 56%) as an orange solid.

[0363] Step 4: Before adding XPhos Pd G3 (74 mg, 0.09 mmol, 0.2 equivalents), ZnCN2 (166 mg, 1.41 mmol, 3 equivalents) was added to a mixture of 2-amino-6-(4-tert-butylphenyl)-5-iodo-4-methylpyridine-3-carboxylate methyl (200 mg, 0.47 mmol, 1 equivalent) in N2-degassed DMA (4.7 mL), and the N2 was degassed again. 120 o After stirring in 14C for 2 hours, the reaction was quenched with NaHCO3 (10 mL), diluted, and extracted with siRNA (3 × 50 mL). The organic layer was washed with brine, dried over MgSO4, filtered through Celite, and concentrated under reduced pressure to obtain methyl 2-amino-6-(4-tert-butylphenyl)-5-cyano-4-methylpyridine-3-carboxylate (104 mg, 0.32 mmol, yield 68%) as an orange solid. The crude product was used in the next step without further purification.

[0364] Step 5: Add a 1M aqueous solution of LiOH (3.22 mL, 3.22 mmol, 10 equivalents) to a solution of methyl 2-amino-6-(4-tert-butylphenyl)-5-cyano-4-methylpyridine-3-carboxylate (104 mg, 0.32 mmol, 1 equivalent) in THF (3.2 mL), and prepare reaction mixture 22. o The mixture was stirred in 14C for 5 hours. The reaction was extracted with saturated aqueous solution of NaCl (10 mL), then with ethylacetate (3 x 50 mL). The organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain [2-amino-6-(4-tert-butylphenyl)-5-cyano-4-methylpyridine-3-carbonyl]oxylithium (101 mg, 0.32 mmol, 99% yield) as a yellow solid. The crude product was used in the next step without further purification.

[0365] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with [2-amino-6-(4-tert-butylphenyl)-5-cyano-4-methylpyridine-3-carbonyl]oxylithium. 1 H NMR(400 MHz,DMSO-d6+D2O)δ 8.35(s,1H),7.68(d, J =8.4 Hz,2H),7.52(d, J =8.4 Hz,2H),7.09(d, J =7.8 Hz,1H),6.90(d, J =8.1 Hz,1H),6.73(d, J =10.3 Hz,1H),6.37(s,1H),6.26(s,1H),4.95 - 4.91(m,1H),4.63 - 4.58(m,1H),4.14 - 4.07(m,1H),4.06 - 3.91(m,5H),3.38 - 3.34(m,1H),3.26 - 3.22(m,1H),3.15 - 3.11(m,1H),3.01 - 2.90(m,4H),2.87 - 2.76(m,2H),2.70-2.66(m,1H),2.36(s,3H),2.34-2.32(m,2H),1.30(s,9H),1.22 - 1.14(m,3H).LCMS(Method 5-100 AB,7 min):R T=2.33 min,[M+H] + =1032.6. Example 69 JPEG0007842812000117.jpg67104

[0366] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-(tert-butyl)phenyl)-6-(difluoromethyl)pyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.29(d,J=7.6 Hz,2H),7.48(d,J=8.0 Hz,2H),7.11-6.96(m,1H),6.90-6.80(m,2H),6.79-6.69(m,2H),6.66-6.56(m, 1H),6.53-6.41(m,1H),5.22-5.12(m,1H),4.82-4.80(m,2H),4.43-4.29(m,5H), 3.63-3.53(m,1H),3.49-3.40(m,1H),3.38-3.34(m,2H),3.28-3.24(m,2H),3.2 4-3.12(m,3H),3.06(s,3H),3.01-2.90(m,2H),1.43-1.28(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.825 min,[M+H] + =1044.4. Example 70 JPEG0007842812000118.jpg67105

[0367] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-amino-2-(4-hydroxy-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)-6-methylpyrimidine-5-carboxylic acid, which was prepared using the procedure described in Example 52. 1H NMR(400MHz,MeOH-d4)δ(ppm)8.41(br s,1H),8.2(d,J=8.4 Hz,1H),7.05(br s,1H),6.93-6.73(m,3H),6.7(d,J=8 Hz,1H),6.59(s,1H),6.5(br s,1H),5.23-5.13(m,1H),4.8(br s,1H),4.43(br s,1H),4.27-3.97(m,6H),3.66-3.54(m,1H),3.41-3.32(m,1H),3.28-2 .98(m,9H),2.89-2.81(m,2H),2.45(s,3H),2.00-1.91(m,2H),1.36(br d,J=6.8 Hz,3H),1.26(s,6H).LCMS(Method 5-95 AB,ESI):R T =0.795 min,[M+H] + =1036.5. Example 71 JPEG0007842812000119.jpg67102

[0368] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-(tert-butyl)phenyl)pyrimidine-5-carboxylic acid. 1 H NMR(400MHz,MeOH-d4)δ(ppm)9.21-9.13(m,2H),8.47-8.25(m,3H),7.55-7.49(m,2H),7. 10-6.35(m,5H),4.30-3.37(m,10H),3.25-2.74(m,8H),1.39-1.36(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.814 min,[M+H] + =979.6. Example 72 JPEG0007842812000120.jpg69107

[0369] Step 1: Copper(II) acetylacetonate (237 mg, 0.905 mmol) was added to a solution of tert-butyl acetoacetate (3.0 mL, 18.1 mmol) and ethyl cyanoformate (1.8 mL, 18.2 mmol) in DCM (12.1 mL). The reaction mixture was stirred at the same temperature for 48 hours and then concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-60% ethyl(E) in heptane) to obtain O1-tert-butylO4-ethyl(E)-2-acetyl-3-aminobuta-2-endioate (1.73 g, yield 37.2%) as a dark gray oily substance (a mixture of two geometric isomers).

[0370] Step 2: 1,1,3,3-tetramethylguanidine (1.69 mL, 13.5 mmol) in DCM (13.4 mL) was added to a solution of O1-tert-butyl O4-ethyl(E)-2-acetyl-3-amino-buto-2-endioate (1.73 g, 6.72 mmol) and 4-tert-butylbenzamidine (1.30 g, 7.40 mmol). The reaction mixture was stirred at room temperature for 16 hours, then quenched with 1 M KHSO4 aqueous solution. The mixture was extracted with ELISA (3×). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% MeOH in DCM) to obtain the title compound 5-tert-butoxycarbonyl-2-(4-tert-butylphenyl)-6-methylpyrimidine-4-carboxylic acid (1.21 g, 48.6% yield) as a grayish-white solid.

[0371] Step 3: Isobutyl chloroformate (501 uL, 3.86 mmol) was added at 0°C to 5-tert-butoxycarbonyl-2-(4-tert-butylphenyl)-6-methylpyrimidine-4-carboxylic acid (1.30 g, 3.51 mmol) and triethylamine (538 uL, 3.86 mmol) in THF (35 mL). The reaction mixture was stirred at room temperature for 15 minutes and then cooled to 0°C. Lithium borohydride (255 mg, 12.3 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour and 30 minutes. MeOH (10 mL) was added at 0°C, followed by saturated NH4Cl aqueous solution. The mixture was then warmed to room temperature and extracted with Â(3×). The organic layers were combined, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 100-200 mesh, 0-10% MeOH in DCM) to obtain tert-butyl 2-(4-tert-butylphenyl)-4-(hydroxymethyl)-6-methylpyrimidine-5-carboxylate (679 mg, yield 54.3%) as a yellow oily substance.

[0372] Step 4: Carbon tetrabromide (444 mg, 1.34 mmol), triphenylphosphine (367 mg, 1.40 mmol), and sodium azide (396 mg, 6.09 mmol) were added to tert-butyl 2-(4-tert-butylphenyl)-4-hydroxymethyl)-6-methylpyrimidine-5-carboxylate (434 mg, 1.22 mmol) in DMF (8.1 mL). Next, triethylamine (373 μL, 2.68 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. The additional portions of carbon tetrabromide (444 mg, 1.34 mmol), triphenylphosphine (367 mg, 1.40 mmol), and triethylamine (373 μL, 2.68 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with ethyl acetate and then washed with saturated aqueous NaHCO3, water, and brine. The organic layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by column chromatography (silica gel, 100-200 mesh, 0-20% ethyl phosphate in heptane) to obtain tert-butyl 4-(azidomethyl)-2-(4-tert-butylphenyl)-6-methylpyrimidine-5-carboxylate (315 mg, yield 67.8%) as a yellow oil.

[0373] Step 5: Fmoc chloride (384 mg, 1.49 mmol) and sodium carbonate (182 mg, 1.71 mmol) were added to a solution of tert-butyl 4-(azidomethyl)-2-(4-tert-butylphenyl)-6-methylpyrimidine-5-carboxylate (436 mg, 1.14 mmol) in ethanol (8.7 mL). The reaction mixture was purged with N2 for 5 minutes, and then palladium carbon (10% load) 55% wet (122 mg, 0.114 mmol) was added. The reaction mixture was purged with H2 for 5 minutes, and then stirred under H2 (1 atm) for 16 hours. The reaction mixture was purged with N2. The additional sodium carbonate (121 mg, 1.14 mmol) and Fmoc chloride (147 mg, 0.568 mmol) were added. The reaction mixture was stirred at room temperature for 5 hours, and then filtered through Celite. The filter cake was washed with MeOH, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel, 100-200 mesh, 0-30% ethyl phosphate in heptane) to obtain tert-butyl 2-(4-tert-butylphenyl)-4-[(9H-fluoren-9-ylmethoxycarbonylamino)methyl]-6-methylpyrimidine-5-carboxylate (487 mg, yield 73.8%) as a colorless oil.

[0374] Step 6: Trifluoroacetic acid (6.5 mL) was added to a solution of tert-butyl 2-(4-tert-butylphenyl)-4-[(9H-fluoren-9-ylmethoxycarbonylamino)methyl]-6-methylpyrimidine-5-carboxylate (487 mg, 0.843 mmol) in DCM (13 mL). The reaction mixture was stirred at room temperature for 12 hours and then concentrated under reduced pressure. The crude product was purified by column chromatography (C-18, 5-70% acetonitrile in 10 mM NH4HCO2 aqueous solution) to obtain 2-(4-tert-butylphenyl)-4-[(9H-fluoren-9-ylmethoxycarbonylamino)-methyl]-6-methylpyrimidine-5-carboxylic acid (303 mg, yield 68.9%) as a grayish-white solid.

[0375] The title compound was prepared as described in Example 1 by replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 2-(4-tert-butylphenyl)-4-[(9H-fluoren-9-ylmethoxycarbonylamino)-methyl]-6-methylpyrimidine-5-carboxylic acid. 1 H NMR(400MHz,DMSO+H2O)δ 8.40(d,J=8.4Hz,2H),8.34(s,2H),7.55(d,J=7.8Hz,2H),7.09 - 7.00(m,1H),6.93 - 6.85(m,1H),6.77 - 6.71(m,1H),6.68(s,1H),6.38(s,1H),6.26(s,1H),5.13 - 5.05(m,1H),4.62(q,J=5.6Hz,1H),4.18 - 3.85(m,9H),3.39 - 3.30(m,2H),3.28 - 3.18(m,1H),3.18 - 3.09(m,1H),3.08 - 2.75(m,7H),2.55(s,3H),1.32(s,9H),1.17(d,J=6.7Hz,3H).LCMS(Method 5-100 AB,7 min):R T =1.46 min,[M+H] + =1022.5. Example 73 JPEG0007842812000121.jpg62102

[0376] The title compound was prepared using the procedure of Example 1, by replacing compound 10 of Example 17 with compound 10 described in step C, replacing tert-butyl(5R)-5-[(3-nitrophenyl)sulfonyloxymethyl]-2-oxo-oxazolidine-3-carboxylate in step 3 with tert-butyl(5S)-5-[(3-nitrophenyl)sulfonyloxymethyl]-2-oxo-oxazolidine-3-carboxylate, and replacing 1-(4-(tert-butyl)phenyl)-6-oxo-1,6-dihydropyridazine-4-carboxylic acid with 4-methyl-2-(4-(1-methylcyclopropyl)phenyl)pyrimidine-5-carboxylic acid prepared as described in Example 5. 1H NMR(400MHz,DMSO+D2O)δ 8.91(d,J=7.1Hz,1H),8.79(s,1H),8.30(d,J=8.5Hz,2H),7.36(d,J=8.6Hz,2H),7.20(d,J= 8.0Hz,1H),7.04(d,J=8.5Hz,1H),6.74(s,1H),6.72(s,1H),6.31(s,1H),6.25(s,1H),4.95 - 4.88(m,1H),4.74 - 4.61(m,2H),4.14 - 4.05(m,1H),4.05 - 3.95(m,4H),3.92 - 3.85(m,1H),3.33 - 3.25(m,1H),3.13 - 2.75(m,10H),2.62(s,3H),2.38(s,15H),2.12 - 1.92(m,2H),1.42(s,3H),1.18(d,J=6.7Hz,3H),0.96 - 0.89(m,2H),0.88 - 0.82(m,2H).LCMS(Method 5-100 AB,7 min):R T =1.45 min, [M+H] + =926.6. Example 74 JPEG0007842812000122.jpg62102

[0377] The general rule of thumb is to use して and the title compound to prepare した. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.83(s,1H),8.35(d,J=8.4 Hz,2H),7.38(d,J=8.4 Hz,2H),7.20-7.14(m,1H),7.03-6.91(m,2H),6.82(s,1H),6.61(s,1H),6.47(s,1H),5.18-5.06(m,1H),4.84-4.76(m,2H),4.61- 4.50(m,1H),4.35-3.92(m,6H),3.60(s,3H),3.26-3.00(m,10H),2.77-2.67(m,3H),2.38-2.12(m,2H),1.47(s,3H),1.37(d,J=7.2 Hz,3H),0.97-0.91(m,2H),0.89-0.81(m,2H).LCMS(Method 5-95 AB, ESI):R T=0.764 min,[M+H] + =940.4. Example 75 JPEG0007842812000123.jpg67102

[0378] The title compound was prepared using general procedure B. 1 H NMR(400MHz,MeOH-d4)δ(ppm)8.46(br s,1H),8.21(d,J=8.0 Hz,2H),7.50(d,J=8.0 Hz,2H),7.40-7.20(m,1H),7.11(d,J=7.6 Hz,1H),6.91-6.83(m,1H),6.80(s,1H),6.63(s,1H),6.57(s,1H),5.24-5.17(m,1H),4. 85-4.78(m,2H),4.46-4.39(m,1H),4.38-4.28(m,1H),4.27-4.20(m,2H),4.19-4.10(m,2 H),4.09-3.94(m,3H),3.70-3.56(m,2H),3.55-3.45(m,3H),3.43-3.34(m,1H),3.25-3. 12(m,2H),3.10(s,3H),3.08-2.94(m,3H),2.49(s,3H),1.41-1.31(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.636 min,[M+H] + =1022.6. Example 76 JPEG0007842812000124.jpg69102

[0379] The title compound was prepared using general procedure B. 1H NMR(400MHz,MeOH-d4)δ(ppm)8.34(br s,1H),8.20(d,J=8.8 Hz,2H),7.50(d,J=8.4 Hz,2H),7.12(d,J=9.2 Hz,1H),6.94-6.79(m,3H),6.60(s,1H),6.50(s,1H),5.15-5.03(m,1H),4.83-4.72(m,1H),4.52-4.42(m,1H),4.26- 4.02(m,6H),3.28-3.00(m,11H),2.47(s,3H),2.23-2.06(m,1H),2.03-1.85(m,1H),1.39-1.34(m,12H).LCMS(Method 5-95 AB,ESI):R T =0.767 min,[M+H] + =1022.6.

[0380] Biotechnology Example B1: LepBアッセイ The in vitro antimicrobial activity of each compound was determined by measuring the minimum inhibitor concentration (MIC) using the broth microdilution technique approved by the Clinical and Laboratory Standards Institute (CLSI) (Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard-Eighth Edition. CLSI document M07-A8. Wayne, PA: Clinical and Laboratory Standards; 2009). Antimicrobial activity was measured against Escherichia coli strain ATCC25922 (E. coli), Klebsiella pneumoniae ATCC700603 (K. pneumoniae), Acinetobacter baumannii ATCC17978 (A. baumannii), and Pseudomonas aeruginosa PA01 (P. aeruginosa), each of which is representative of clinically relevant Gram-negative bacteria. Cells were seeded on Mueller-Hinton agar plates and grown at 37°C for 16-18 hours. Inoculum suspensions were prepared by scraping cells into 1 mL of test medium (Müller-Hinton II cation-adjusted broth) and diluting to a final OD of 0.01 at 600 nm.

[0381] The test compound was prepared in DMSO at a concentration of 64 ug / ml. The compound was tested in several different dilution forms. In Protocol 1, the compound stock was diluted in test medium at a concentration of 64 μg / ml, and a 2-fold serial dilution was performed in the same medium in a 96-well U-bottom microtiter dish to obtain a total of 10 compound concentrations. In Protocol 2, the compound stock was diluted in test medium at a concentration of 4 μg / mL, and a 2-fold serial dilution was performed in the same medium in a 96-well U-bottom microtiter dish to obtain a total of 10 compound concentrations. In Protocol 3, the compound stock was diluted in test medium at a concentration of 0.5 μg / mL, and a 2-fold serial dilution was performed as described above. In Protocol 4, the compound stock was diluted in test medium at a concentration of 0.13 μg / mL, and a 2-fold serial dilution was performed as described above. The inoculum suspension was added to the 2-fold serial dilution of the test compound to a final density of 0.0005 OD600 nm, and incubated at 35°C for 22 hours. After incubation, the plates were visually inspected, and the lowest concentration of the test compound that completely inhibited bacterial growth was recorded as the MIC. To evaluate antimicrobial activity in the presence of plasma proteins, the MIC was also measured in growth medium supplemented with 50% serum. Bacterial inoculum and diluted test compound plates were prepared using the same protocol as described above for standard MIC assays, except that a mixture of 50% v / v Mueller-Hinton II cation-modified broth and 50% v / v sterile mouse filtered serum (Equitech-Bio) was used instead of 100% Mueller-Hinton II cation-modified broth. The results are shown in Table 3 (MIC, IC). 50 The value is in micromoles.

[0382] Example B2: Measurement of oxygen consumption Isolation of liver mitochondria Male Sprague-Dawley rats (8-10 weeks old) were euthanized by carbon dioxide overdose. The livers were rapidly excised and placed in ice-cold phosphate-buffered saline (PBS) before the isolation process. Mitochondria were isolated using the MitoCheck® mitochondrial isolation kit (Cayman Chemical #701010, Ann Arbor, Michigan) according to the manufacturer's instructions. Briefly, approximately 10-14 grams of liver tissue were finely chopped with scissors and repeatedly washed with ice-cold PBS, followed by Mito isolation buffer. The chopped tissue was homogenized in 30 ml of Mito homogenization buffer in a 40 ml dance-type homogenizer and a smooth glass grinder. The homogenate was divided into two clean 30 ml polycarbonate tubes and centrifuged at 1,000 × g at 4°C for 3 minutes. The supernatant was further centrifuged at 10,000 × g at 4°C for 10 minutes. The supernatant was discarded, and the mitochondrial pellet was suspended in Mito isolation buffer using a glass stirring rod and stored on ice until ready for use. Mitochondrial protein concentrations were determined using a bicinchoninate protein assay kit (Thermo Scientific, Waltham, Massachusetts).

[0383] Measurement of oxygen consumption Oxygen consumption was monitored as essentially described by Will et al., “Analysis of mitochondrial function using phosphorescent oxygen-sensitive probes,” Nature Protocols Vol.1(6)(2006)2563. Briefly, isolated mitochondria (50 μg mitochondrial protein / state 3 (S3) for mitochondrial respiration identification, 100 μg / S2 for uncoupling agent identification) were incubated with the compounds on ice for 1 hour in 25 μl of measurement buffer containing 1% DMSO (MB250 mM sucrose, 15 mM KCl, 1 mM EGTA, 5 mM MgCl2, 30 mM K2HPO4, pH 7.4), and then tested with an oxygen-sensitive fluorescent dye (MitoXpress). 25 μl of the substrate mixture was mixed in a 384-well plate with black walls and a clear bottom containing Xtra (Agilent, Santa Clara, California), glutamate (25 mM, G1626, Sigma, St. Louis, Missouri), malate (25 mM, M6413, Sigma), ADP (3.3 mM, A2754, Sigma, only for S3 in mitochondrial respiration), fatty acid-free BSA (0.1%, A0281, Sigma), and the complex II-specific inhibitor TTFA (2 μM, T27006, Sigma, for evaluation of complex I-mitochondrial respiration with glutamate / malate). The mixture was then coated with HS mineral oil (Agilent) using a Viaflo 384 channel pipette (Integra, Hudson, New Hampshire). Oxygen consumption was measured for 30 minutes at 30°C using a spectrofluorometer (FLUOstar Omega, BMG Labtech, Carey, North Carolina). In S3, 0% and 100% inhibition were defined using DMSO vehicle control and antimycin A (10 μM). In S2, 0% and 100% uncoupling were defined using DMSO vehicle and FCCP (0.5–1 μM), respectively. The area under the curve, typically in the range of 0–12 minutes (S3) or 0–24 minutes (S2), was used for calculations. IC 50 or UC 50The (uncoupling concentration) was the concentration that caused 50% inhibition of oxygen consumption or maximal (uncoupling) activation, and was used to assess the quality of the experiment. Data were expressed as μM (per 50 μg (S3) or 100 μg (S2) of mitochondrial protein). IC 50 <100 μM (S3) and / or UC 50 Compounds with <200 μM (S2) were considered to have a high risk of mitochondrial toxicity. Compounds showing a 25-50% change in inhibition and / or uncoupling were classified as medium risk, as compounds showing a change greater than 25% were considered responders.

[0384] Toxicity associated with mitochondrial dysfunction, as with other types of therapeutic agents, is a concern for promising potential antibiotics. Mitochondrial oxygen con...

Claims

1. The following compounds: A compound selected from the group consisting of the following, or a salt thereof.

2. The following structure: A compound having, or a salt thereof.

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