Novel compounds for the treatment of bacterial infections

Abstract

The present invention relates to formula (I) With respect to the compound of TIFF2026514062000228.tif7975, in the formula, R1 to R7, n, X, L, A1 and A2 are as defined herein and in the claims. The compound of formula (I) can be used as a pharmaceutical product.

Description

[Technical Field]

[0001] This invention relates to organic compounds useful for treating and / or preventing bacterial infections in mammals. Specifically, these molecules can inhibit the LPS synthesis pathway, and in particular LpxH, and are useful for treating bacterial infections.

[0002] The present invention particularly relates to formula (I) [ka] (In the formula, L is either absent or alkyl. X is -O- or -NH-, n is either 0 or 1. R1 is amino, aminoalkyl, alkylamino, aminoalkylamino, haloalkyl, diaminoalkyl, guanidinyl, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkyl, aminoalkoxyalkyl, (aminoalkyl)(aminoalkyl)(aminocarbonylalkyl)ammoniumalkyl, (amino)(hydroxy)alkyl, (aminoalkyl)(aminoalkyl)amino, (aminoalkyl)(aminoalkyl)aminoalkyl, (aminoalkyl)(aminoalkyl)aminoalkoxy, (aminoalkyl)(aminoalkyl)aminoalkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonyl)ammoniumalkylamino, (hydroxy)(halo)alkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkoxy, cycloalkyl, or heterocycloalkyl, where heterocycloalkyl and cycloalkyl are optionally substituted with one or two substituents independently selected from R6. However, if R1 is an amino acid, L does not exist. R2 and R3 are independently selected from halogens, or R2 and R3 together with the carbon to which they are bonded form a cycloalkyl group. R4 is a halogen or alkyl, R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino, or piperazinyl. In each example, R6 is independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl, and aminoalkylcarbonylamino. R7 is hydrogen or hydroxyl. A1 is -N- or -CH-, A2 is a compound (which is -O- or -CH2-), or relating to a pharmaceutically acceptable salt thereof. [Background technology]

[0003] The intensive use of antibiotics has exerted selective evolutionary pressure on microorganisms to create gene-based resistance mechanisms. Modern medical and socioeconomic practices exacerbate the problem of resistance development, for example by creating conditions that allow pathogenic microorganisms to grow slowly within artificial joints, and by supporting long-term host reservoirs in immunocompromised patients. In hospital settings, the number of major infectious agents such as Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus spp., and Enterobacteriaceae (e.g., Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa) is increasing, becoming multidrug resistant, and thus becoming difficult to treat. This is especially true for Gram-negative organisms, where the situation is becoming increasingly concerning as no novel drugs with differentiated mechanisms of action have been approved for decades. Therefore, there is a critical medical need for novel antimicrobial compounds to address Gram-negative resistant bacteria, particularly third-generation cephalosporin and carbapenem-resistant Enterobacteriaceae, as well as multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumanni. One way to address the problem of cross-resistance to established classes of antibiotics is to inhibit essential proteins or functions not targeted by current antibiotics.

[0004] Gram-negative bacteria are unique in that their outer membrane contains lipopolysaccharide (LPS), which is crucial for maintaining membrane integrity and essential for bacterial survival (revisited in Ann. Rev. Bi℃hem 76:295-329, 2007). The main lipid component of LPS is lipid A, and inhibition of lipid A biosynthesis is lethal to bacteria. Lipid A is synthesized on the cytoplasmic surface of the bacterial inner membrane via a pathway consisting of nine different enzymes. These enzymes are highly conserved in most Gram-negative bacteria. LpxH, a calcineurin-like phosphatase (CLP), catalyzes the hydrolysis of UDP-2,3-diacyl-glucosamine (UDP-DAGn) to produce lipids X and UMP. LpxH has no mammalian homolog and is a good target for the development of novel antibiotics targeting Gram-negative bacteria. [Overview of the project]

[0005] The term "alkyl" alone or in combination refers to a monovalent linear or branched saturated hydrocarbon group with 1 to 12 carbon atoms, particularly 1 to 7 carbon atoms, and more specifically 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl. Methyl, ethyl, and propyl are specific examples of "alkyl".

[0006] The term "cycloalkyl" alone or in combination refers to a cycloalkyl ring having 3 to 8 carbon atoms, and more particularly a cycloalkyl ring having 3 to 6 carbon atoms. Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Specific examples of "cycloalkyl" are cyclopropyl and cyclobutyl.

[0007] The term "heterocycloalkyl" refers to a monocyclic, bicyclic, or tricyclic ring system of 4 to 12 ring atoms, particularly 5 to 7 ring atoms, that contains one, two, or three ring heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. A bicyclic ring system consists of two rings sharing one or two ring atoms. A "heterocycloylalkyl" may also contain a carbonyl group, where the carbon of the carbonyl group is part of the ring system. The ring system can be bonded to the rest of the compound via atoms selected from C, N, S, and O, particularly via N atoms ("N-heterocycloalkyl"). Examples of "heterocycloalkyl" include, but are not limited to, morpholino, morpholin-4-yl, pyrrolidinyl, pyrrolidin-1-yl, pyrrolidin-3-yl, piperidinyl, 1-piperidyl, 4-piperidyl, 2-oxopyrrolidin-1-yl, piperazinyl, piperazine-1-yl, azetidinyl, azetidine-1-yl, azetidine-3-yl, azetidine-1-yl, or 5-azoniaspiro[2.4]heptane. Specific examples include azetidine-3-yl, azetidine-1-yl, morpholin-4-yl, pyrrolidin-1-yl, and 5-azoniaspiro[2.4]heptane.

[0008] The terms "alkoxy" or "alkyloxy," alone or in combination, refer to the alkyl-O- group of the formula, while the term "alkyl" refers to the previously given meanings, e.g., methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy. Specific examples of "alkoxy" are methoxy and ethoxy.

[0009] The term "oxy," either alone or in combination, refers to an -O- group.

[0010] The terms "halogen" or "halo," alone or in combination, mean fluorine, chlorine, bromine, or iodine, and in particular fluorine, chlorine, or bromine, more specifically fluorine. The term "halo," in combination with another group, means the substitution of the group with at least one halogen, in particular substituted with 1 to 5 halogens, in particular substituted with 1 to 4 halogens, i.e., 1, 2, 3, or 4 halogens.

[0011] The term "haloalkyl" refers to an alkyl group, either alone or in combination, that is substituted with at least one halogen, particularly 1 to 5 halogens, particularly 1 to 3 halogens, and more specifically 2 to 3 halogens. Specific "haloalkyl" groups include fluoromethyl, fluoroethyl, fluoropropyl, fluorobutyl, difluoromethyl, difluoroethyl, trifluoromethyl, and trifluoroethyl.

[0012] The term "haloalkoxy" refers to an alkoxy group substituted with at least one halogen, particularly 1 to 5 halogens, or especially 1 to 3 halogens, either alone or in combination. Specific "haloalkoxys" include fluoromethoxy, fluoroethoxy, and fluoropropyloxy.

[0013] The terms "hydroxyl" and "hydroxy," alone or in combination, refer to the -OH group.

[0014] The term "carbonyl," either alone or in combination, refers to a -C(O)- group.

[0015] The term "amino," either alone or in combination, refers to a primary amino group (-NH2), a secondary amino group (-NH-), or a tertiary amino group (-N-).

[0016] The term "alkylamino" refers to an alkyl group bonded to an -NH- group, either alone or in combination. The term "dialkylamino" refers to two alkyl groups bonded to an -N- atom.

[0017] The term "aminoalkyl" refers, alone or in combination, to an amino group bonded to an alkyl group, which is further bonded to the compound as shown. The term "diaminoalkyl" refers to two amino groups bonded to an alkyl group, which is further bonded to the compound as shown.

[0018] The term "aminoalkoxy," either alone or in combination, refers to an amino group bonded to an alkoxy group, which is further bonded to the compound via an -O- atom.

[0019] The term "aminoalkoxyalkyl" refers, either alone or in combination, to an amino group bonded to an alkyl group via an alkoxy, where the alkyl group is further bonded to the compound as shown.

[0020] The term "ammonium alkyl," either alone or in combination, refers to an ammonium group bonded to an alkyl group, which is further bonded to the compound as shown.

[0021] The term "alkylcarbonyl" refers to an alkyl group bonded to a -C(O)- group, with a carbonyl group further bonded to the compound as shown. Specific "alkylcarbonyls" include methylcarbonyl (also known as acetyl) and ethylcarbonyl.

[0022] The term "carbonylalkyl" refers to a carbonyl group bonded to an alkyl group, with the alkyl group further bonded to the compound as shown. Specific "alkylcarbonyl" compounds include carbonylmethyl and carbonylethyl.

[0023] The term "carbonylamino" refers to a carbonyl group bonded to an amino group, and the amino group is further bonded to the compound as shown.

[0024] The term "sulfonyl," either alone or in combination, refers to the -SO2- group.

[0025] The term "pharmaceutically acceptable salt" means a salt that is not biologically or otherwise undesirable. Pharmacopoecitable salts include both acid addition salts and base addition salts. The term "pharmaceutically acceptable acid addition salt" refers to a pharmaceutically acceptable salt formed with an organic acid selected from aliphatic, alicyclic, aromatic, aromaticaliphatic, heterocyclic, carboxylic acid, and sulfonic acid derivatives, such as inorganic acids like hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. The term "pharmaceutically acceptable base addition salt" refers to a pharmaceutically acceptable salt formed with an organic or inorganic base. Examples of acceptable inorganic bases include sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, and aluminum salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydravamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperidine, N-ethylpiperidine, and salts of polyamine resins. The term “pharmaceutically acceptable salt” includes, but is not limited to, the salts exemplified herein. The term “pharmaceutically acceptable salt” also includes salts in which the counterion is exchanged with another suitable counterion, in particular a suitable anion.

[0026] The terms "compound(s) of this invention" and "compound(s) of the present invention" refer to the compound of formula (I) and its stereoisomers, tautomers, solvates, and salts (e.g., pharmaceutically acceptable salts).

[0027] Tautomers, that is, structural isomers that interconvert with the compound of formula (I), particularly in solution, may exist in some cases and should be understood to be included in the present invention.

[0028] If one of the starting materials or compounds of formula (I) of the present invention contains one or more functional groups that are unstable or reactive under the reaction conditions of one or more reaction steps, an appropriate protecting group (e.g., “Protective Groups in Organic Chemistry” by TWGreene and PGMWuts, 3) rd Protecting groups (such as those described in Ed., 1999, Wiley, New York) may be introduced before key steps in applying methods well known in the art. Such protecting groups can be removed in later steps of synthesis using standard methods described in the literature. Examples of protecting groups include tert-butoxycarbonyl (B°C), 9-fluorenylmethylcarbamate (Fm°C), 2-trimethylsilylethylcarbamate (Te°C), carbobenzyloxy (Cbz), and p-methoxybenzyloxycarbonyl (Moz).

[0029] The compound of formula (I) may contain several chiral centers and may exist as an optically pure enantiomer, a mixture of enantiomers such as a racemate, a mixture of diastereomers, a diastereomer racemate, or a mixture of diastereomer racemates.

[0030] The term "chiral carbon atom" refers to a carbon atom having four different substituents. According to the Kahn-Ingold-Prelogue priority rule, a chiral carbon atom can have either an "R" or "S" stereoconfiguration.

[0031] Furthermore, the present invention includes, where applicable, all optical isomers of the compounds of formula (I), namely diastereomers, diastereomer mixtures, racemic mixtures, all their corresponding enantiomers and / or tautomers, and their solvated compounds.

[0032] If necessary, the racemic mixture of the compounds of the present invention may be separated to isolate the individual enantiomers. Separation can be carried out by methods known in the art, such as coupling the racemic mixture of compounds enantiomerically with the pure compounds to form a diastereoisomerized mixture, and then separating the individual diastereomers by standard methods such as fractional recrystallization or chromatography.

[0033] In one embodiment, when an optically pure enantiomer is provided, an optically pure enantiomer means that the compound contains more than 90% by weight of the desired isomer, more than 95% by weight of the particularly desired isomer, or more specifically, more than 99% by weight of the desired isomer, where such weight percentage is based on the total weight of the isomers of the compound. Chiralally pure or chiralally concentrated compounds can be prepared by chiral selective synthesis or by separation of enantiomers. Separation of enantiomers can be performed on the final product or on a suitable intermediate.

[0034] The structures shown herein also mean that they include compounds that differ only in the presence of one or more isotopically enriched atoms. Specific examples of radioactive isotopes are: 2 H, 3 H, 13 C, 14 C and 18 It is F. For example, a structure in which one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms13 C or 14 Structures in which carbon is replaced with 1C-enriched carbon are within the scope of the present invention.

[0035] Therefore, the present invention relates to the following numbered embodiments.

[0036] 1. Equation (I) [ka] (In the formula, L is either absent or alkyl. X is -O- or -NH-, n is either 0 or 1. R1 is amino, aminoalkyl, alkylamino, aminoalkylamino, haloalkyl, diaminoalkyl, guanidinyl, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkyl, aminoalkoxyalkyl, (aminoalkyl)(aminoalkyl)(aminocarbonylalkyl)ammoniumalkyl, (amino)(hydroxy)alkyl, (aminoalkyl)(aminoalkyl)amino, (aminoalkyl)(aminoalkyl)aminoalkyl, (aminoalkyl)(aminoalkyl)aminoalkoxy, (aminoalkyl)(aminoalkyl)aminoalkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonyl)ammoniumalkylamino, (hydroxy)(halo)alkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkoxy, cycloalkyl, or heterocycloalkyl, where heterocycloalkyl and cycloalkyl are optionally substituted with one or two substituents independently selected from R6. However, if R1 is an amino acid, L does not exist. R2 and R3 are independently selected from halogens, or R2 and R3 together with the carbon to which they are bonded form a cycloalkyl group. R4 is a halogen or alkyl, R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino, or piperazinyl. In each example, R6 is independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl, and aminoalkylcarbonylamino. R7 is hydrogen or hydroxyl. A1 is -N- or -CH-, A2 is a compound (which is -O- or -CH2-), or a pharmaceutically acceptable salt thereof.

[0037] 2. The compound according to embodiment 1, wherein L is absent.

[0038] 3. The compound according to embodiment 1 or 2, wherein L is alkyl, particularly methyl.

[0039] 4. A compound according to any one of embodiments 1 to 3, wherein n is 1.

[0040] 5. A compound according to any one of embodiments 1 to 4, wherein X is -O-.

[0041] 6. The compound according to any one of embodiments 1 to 5, wherein X is -NH-.

[0042] 7. A compound according to any one of embodiments 1 to 3, wherein n is 0.

[0043] 8. R1 is amino, aminomethyl, aminoethyl, methylamino, aminoethylamino, aminopropylamino, trifluoromethyl, diaminoethyl, guanidinyl, aminoethoxymethyl, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropyl, (aminopropyl)(aminopropyl)(aminocarbonylmethyl)ammoniumpropyl, (amino)(hydroxy)ethyl, (aminopropyl)(aminopropyl)amino, (aminoethyl)(aminoethyl)aminopropyl, (aminopropyl)(aminopropyl)aminopropyl, (aminopropyl)(aminopropyl)aminoethoxy, (aminoethyl)(aminoethyl)aminoethoxy, (aminoethyl)(aminoethyl) Minoethylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropylamino, (hydroxy)difluoropropylamino, (hydroxy)trifluoropropylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumethoxy, cyclobutyl, azetidine-3-yl, azetidine-1-yl, morpholine-4-yl, pyrrolidine-1-yl, or 5-azoniaspiro[2.4]heptane, wherein cyclobutyl, azetidine-3-yl, azetidine-1-yl, morpholine-4-yl, pyrrolidine-1-yl, or 5-azoniaspiro[2.4]heptane is optionally substituted with one or two substituents independently selected from R6. However, if R1 is an amino acid, L is absent, the compound according to any one of embodiments 1 to 7.

[0044] 9. The compound according to any one of embodiments 1 to 7, wherein R1 is a heterocycloalkyl group in which R1 is optionally substituted with one or two substituents independently selected from R6.

[0045] 10. The compound according to any one of embodiments 1 to 9, wherein both R2 and R3 are fluorides, or R2 and R3, together with the carbons to which they are bonded, form a cyclopropyl group.

[0046] 11. The compound according to any one of embodiments 1 to 10, wherein R4 is chloride or methyl, particularly chloride.

[0047] 12. The compound according to any one of embodiments 1 to 10, wherein R4 is methyl.

[0048] 13. The compound according to any one of embodiments 1 to 12, wherein R5 is amino or hydroxyl.

[0049] 14. The compound according to any one of embodiments 1 to 12, wherein R5 is an amino acid.

[0050] 15. The compound according to any one of embodiments 1 to 14, wherein R6 is independently selected from amino, methyl, hydroxy, aminomethyl, aminocarbonylmethyl, and aminoethylcarbonylamino in each example.

[0051] 16. A compound according to any one of embodiments 1 to 15, wherein R7 is hydrogen.

[0052] 17. The compound according to any one of embodiments 1 to 16, wherein A1 is -CH-.

[0053] 18. The compound according to any one of embodiments 1 to 17, wherein A2 is -CH2-.

[0054] 19. A compound selected from the following: cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]azetidine-3-yl]propanamide; (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; trans-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide; cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-hydroxy-propanamide; cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]urea; trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5,5-dimethyl-5-azoniaspiro[2,4]heptan-6-carboxamide; trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5-methyl-5-azoniaspiro[2,4]heptan-6-carboxamide; trans-(5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5-methyl-5-azoniaspiro[2,4]heptan-6-carboxamide; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; 2-Amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]acetamide; 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N-(carboxymethyl)-4-oxobutan-1-aminium; N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-(2-aminoethoxy)acetamide; 3-Amino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-hydroxypropanamide; 2,3-Diamino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)propanamide; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(diaminomethylene)urea; (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]amino]-4-oxo-butyl]-bis(3-aminopropyl)ammonium; 3-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoylamino]propyl-bis(3-aminopropyl)-(carboxymethyl)ammonium; 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(3-aminopropyl)guanidine; N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]-2,2,2-trifluoroacetamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)-(carboxymethyl)ammonium; (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]morpholine-4-carboxamide; N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N-hydroxybutanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxypropyl)amino]cyclohexyl]-difluoromethyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-hydroxy-pyrrolidine-1-carboxamide; 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2-aminoethyl)amino)ethyl)urea; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-methylpyrimidine-2-yl]piperazin-1-yl]sulfonylphenyl]pyrrolidine-2-one; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-methylpyrimidine-4-yl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[[4-(azetidine-1-yl)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; tert-butyl((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate; or a pharmaceutically acceptable salt thereof.

[0055] 20. A compound according to any one of embodiments 1 to 19, for use as a therapeutically active substance.

[0056] 21. A pharmaceutical composition comprising a compound described in any one of embodiments 1 to 19 and a therapeutically inert carrier.

[0057] 22. Use of the compound according to any one of embodiments 1 to 19 for the treatment or prevention of bacterial infections, particularly bacterial infections in which the bacteria are Gram-negative bacteria.

[0058] 23. Use of the compound according to any one of embodiments 1 to 19 for the preparation of a pharmaceutical product for the treatment or prevention of a bacterial infection, particularly a bacterial infection in which the bacteria are Gram-negative bacteria.

[0059] 24. A compound according to any one of embodiments 1 to 19 for use in the treatment or prevention of bacterial infections, particularly bacterial infections in which the bacteria are Gram-negative bacteria.

[0060] 25. A method for the treatment or prevention of a bacterial infection, particularly an infection of Gram-negative bacteria, comprising administering an effective amount of any one of embodiments 1 to 19.

[0061] 26. The use, method, or compound for use according to any one of aspects 22 to 25, wherein the Gram-negative bacterium is selected from Enterobacteriaceae, Neisseria gonorrhoeae, Haemophilus influenzae, Helicobacter pylorus, Acinetobacter baumannii, and Pseudomonas aeruginosa.

[0062] 27. The use, method, or compound for use according to aspect 26, wherein the Gram-negative bacterium is Enterobacteriaceae, and Enterobacteriaceae is Klebsiella pneumoniae or Escherichia coli. One embodiment of the present invention is that the compound is a compound of formula (II)

Chemical formula

Mode for Carrying Out the Invention

[0063] The synthesis of the compound of formula (I) can be achieved, for example, according to the non-exhaustive procedures described below in General Schemes 1 to 12, or according to methods known to those skilled in the art. In some cases, the order of the reaction steps can be changed, and the individual steps of different schemes can be combined as disclosed herein. All substituents are as defined in the specification and claims, unless otherwise indicated. Further, unless otherwise specified, all reactions, reaction conditions, abbreviations, and symbols have meanings generally known to those skilled in the art.

[0064] Scheme 1

Chemical formula

[0065] The compound of formula (IV) can be obtained from a sulfonation reaction between nitrophenylsulfonyl chloride (II) and the compound of formula (III). The reduction of the nitro group in the compound of formula (IV) to aniline (V) can be carried out by using platinum on carbon in the presence of H2. Amidation of the compound of formula (V) with lactone (VI) in the presence of a base such as LHMDS yields amide (VII). Cyclization of the primary alcohol (VII) via the reaction yields a cyclic lactam (VIII). The compound of formula (IX) can be obtained by deprotecting the compound of formula (VIII) with a reducing agent such as palladium-carbon in the presence of H2.

[0066] Scheme 2 [ka] In the above scheme, B 1 is halogen or -SMe, B 2 is halogen or H, R4 is halogen, A1 is -N- or -CH-, PG 3 and PG 4 These are independently selected protecting groups, such as tert-butoxycarbonyl and 2-(trimethylsilyl)ethoxycarbonyl groups.

[0067] Compound (XII) can be obtained by magnesium-halogen exchange or deprotonation of the compound of formula (X) with a suitable Grignard reagent, such as isopropylmagnesium chloride lithium chloride complex or chlorolithium;chloro-(2,2,6,6-tetramethyl-1-piperidyl)magnesium, followed by the addition of compound (XI). Compound (XIII) can be obtained by oxidation of compound (XII) in the presence of a suitable oxidizing agent such as Dess Martin's reagent. Compound (XIV) can be obtained by fluorination of compound (XIII) in the presence of a suitable reagent such as DAST. Compound (XV) can be obtained by deprotecting compound (XIV) with a suitable acid, such as trifluoroacetic acid. Compound (XVI) can be obtained by protection of amine (XV) at PG4 with 1-[2-(trimethylsilyl)ethoxycarbonyloxy]pyrrolidine-2,5-dione (Te℃-OSu).

[0068] Scheme 3 [ka] In the above scheme, B 1 is halogen or -SMe, B 2 is a halogen or H, R4 is a halogen, A1 is -N- or -CH-, R is alkyl, PG 3 This is a protecting group such as the tert-butoxycarbonyl group.

[0069] The compound of formula (XVII) can be obtained by alkylating the compound of formula (XIII) with an alkyl halide, such as MeI, and a base, such as NaH. The compound of formula (XVIII) can be obtained by deprotecting the compound of formula (XVII) with a suitable acid, such as trifluoroacetic acid.

[0070] Scheme 4 [ka] In the above scheme, B 1is halogen or -SMe, B 2 is a halogen or H, R4 is a halogen, and A1 is -N- or -CH-.

[0071] Compound (XX) can be obtained by magnesium-halogen exchange or deprotonation of the compound of formula ((X)) with a suitable Grignard reagent, such as isopropylmagnesium chloride lithium chloride complex or chlorolithium; followed by the addition of chloro-(2,2,6,6-tetramethyl-1-piperidyl)magnesium and then the addition of the compound of formula (XIX). Compound (XX) can be obtained by oxidation of the compound of formula (XX) in the presence of a suitable oxidizing agent such as Dess Martin's reagent. Compound (XXII) can be obtained by fluorination of the compound of formula (XXI) in the presence of a suitable reagent such as DAST. Compound (XXIII) can be obtained by reduction of compound (XXII) with a suitable reducing agent, such as lithium aluminum hydride.

[0072] Scheme 5 [ka] In the above scheme, B 1 is halogen or -SMe, B 3 is -SO2Me, R4 is halogen, and R 4a A1 is alkyl, A1 is -N- or -CH-, and PG 5 This is a protecting group, such as a benzyloxycarbonyl group.

[0073] The compound of formula (XXIV) can be obtained by Suzuki coupling of the compound of formula (XXII) with an alkylboronic acid or ester catalyst, such as Pd-XPhos-G4, and a base, such as K3PO4. The compound of formula (XXV) can be obtained by hydrolysis of the compound of formula (XXIV) under suitable conditions such as LiBr and Et3N. The compound of formula (XXVI) can be obtained by Curtius rearrangement of the compound of formula (XXV) in the presence of DPPA and Et3N, followed by capture of the isocyanate with benzyl alcohol. The compound of formula (XXVII) can be obtained by oxidation of the compound of formula (XXVI) in the presence of a suitable oxidizing agent such as mCPBA.

[0074] Scheme 6 [ka] In the above scheme, B 1 R4 is a halogen or -SMe, A1 is -N- or -CH-, X is -O- or -NH-, Y is OH or a halogen and a leaving group such as -O-4-NO2Ph, and PG 3 and PG 6 These are independently protecting groups, such as benzyloxycarbonyl and tert-butoxycarbonyl groups.

[0075] The compound of formula (XXIX) can be obtained by the Wittig reaction of the compound of formula (XIII) using a suitable reagent such as methyltriphenylphosphonium bromide and a base such as n-BuLi. The compound of formula (XXX) can be obtained by cyclopropanation of the compound of formula (XXIX) under suitable conditions using a base such as NaH in the presence of trimethylsulfoxonium iodide. The compound of formula (XXXI)c can be obtained by deprotecting the compound of formula (XXX) with a suitable acid, such as trifluoroacetic acid. The compound of formula (XXXIII) can be obtained by amidation of the compound of formula (XXXI) with the compound of formula (XXXII) using suitable coupling conditions such as HATU in the presence of a base such as DIEA.

[0076] Scheme 7 [ka] In the above scheme, B 1 is a halogen, B 3 R4 is -SO2Me, R4 is halogen or alkyl, A1 is -N- or -CH-, R7 is H or -OH, R is alkyl or hydroxyalkyl, alkyl halide, alkyl halide alcohol, cycloalkyl together is amine, cycloalkyl alcohol together is amine, PG 2 PG 3 PG 4 and PG 5 These are independently protecting groups such as tert-butoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl, and benzyloxycarbonyl groups.

[0077] Compound (XXXIV) can be obtained by nucleophilic substitution of compounds of formula (XIII, XVI, XXVII) with compound (IX) in the presence of a suitable base such as DIEA. Compound (XXXV) can be obtained by selectively deprotecting compounds of formula (XVI, XXVII) under suitable F anion conditions such as tetraethylammonium fluoride (TEAF), or under reducing conditions such as PdCl2, Et3SiH, or TEA.

[0078] The compounds of formula (XXXVI) can be obtained from the compounds of formula (XXXV) by several different methods: (1) a three-step conversion to -NHOH(R7:OH) by cyanomethylation with cyanomethyl bromide, mCPBA oxidation and subsequent addition of NH2OH; (2) dialkylation of -NH2 with XCH2CH(Z)CH2X to form a four-membered ring; and (3) reductive amination with various aldehydes using a suitable reducing agent such as NaBH(OAc)3 in the presence of AcOH.

[0079] (4) Epoxide ring-opening in the presence of a base such as DIEA. Deprotection in the compounds of formula (XXXV) and (XXXVI) can be performed using a suitable acid, such as trifluoroacetic acid, to give the compounds of formula (XXXVII) and (XXXVIII), respectively.

[0080] Scheme 8 [ka] In the formula, B 1 R4 is a halogen, A1 is -N- or -CH-, R is an alkyl, and PG 2 and PG 3 It is independently a protecting group such as the tert-butoxycarbonyl group.

[0081] The overall deprotection of the compound of formula (XXXIV) can also be performed using a suitable acid such as trifluoroacetic acid to obtain the compound of formula (XXXVII). Alternatively, the compound of formula (XXXVI) can be obtained by substituting the compound of formula (XVIII) with the compound of formula (IX) in the presence of a suitable base such as DIEA.

[0082] Scheme 9 [ka] In the above scheme, R4 is a halogen or alkyl, A1 is -N- or -CH-, X is -O-, R1 is a C or O-linked substituent (amide or carbamate), Y is an OH or halogen and a leaving group such as -O-4-NO2Ph, and PG 2 and PG 6 These are, independently, protecting groups such as tert-butoxycarbonyl, 2-, and benzyloxycarbonyl groups. In some cases, PG 6 There is no protection.

[0083] The compound of formula (XXXIX) can be obtained by two methods: (1) coupling of the compound of formula (XXXV) and the compound of formula (XXXII) using appropriate coupling conditions such as HATU in the presence of a base such as DIEA (C-linked amide); and (2) amidation of the compound of formula (XXXV) and the compound of formula (XXXII). 6 ) is activated by reacting it with 4-nitrophenyl chloroformate to produce YCOO-R1-PG. 6 (XXXII) is produced and reacted with the amine (XXXV) to obtain the carbamate (XXXIX). Deprotection / overall deprotection of the compound of formula (XXXIX) can be performed using a suitable acid such as trifluoroacetic acid to obtain the compound of formula (XXXX).

[0084] Scheme 10 [ka] In the scheme above, R4 is a halogen or alkyl, A1 is -N- or -CH-, X is -O- or -NH, Z is N-imidazole, and PG 2 and PG 7 These are, independently, protecting groups such as tert-butoxycarbonyl, 2-, and benzyloxycarbonyl groups. In some cases, PG 7 There is no protection.

[0085] The compounds of formula (XXXXII) can be obtained by coupling the compound of formula (XXXV) with the compound of formula (XXXXI). N,N'-carbonyldiimidazole (CDI) or imidazole-1-yl(1,2,4-triazole-4-yl)methaneimine is used as the compound of formula (XXXXI). The compounds of formula (XXXXIV) can be obtained by substituting the amine (XXXXIII) with the compound of formula (XXXXII) in the presence or absence of a base such as K2CO3. Deprotection / overall deprotection of the compound of formula (XXXXIV) can be performed using a suitable acid such as trifluoroacetic acid to obtain the compound of formula (XXXXV).

[0086] Scheme 11 [ka] In the above scheme, B 1 R4 is a halide or -SO2Me, R4 is a halogen or alkyl, A1 is -N- or -CH-, X is -O- or -NH, Y is an OH or halogen and a leaving group such as -O-4-NO2Ph, and PG 2 and PG 6 It is independently a protecting group such as the tert-butoxycarbonyl group. In some cases, PG 6 There is no protecting group.

[0087] The compound of formula (XXXXVI) can be obtained by substituting the compound of formula (XXIII) with the compound of formula (IX) in the presence of a suitable base such as DIEA.

[0088] The compound of formula (XXXXVII) can be obtained by converting a primary alcohol (XXXXVI) to a leaving group such as -OTf using Tf2O in the presence of a base such as DIEA, and then substituting it with NH3. The compound of formula (XXXXVIII) can be obtained by amidating the compound of formula (XXXXVII) with the compound of formula (XXXII) using appropriate coupling conditions such as HATU in the presence of a base such as DIEA. Deprotection / overall deprotection of the compound of formula (XXXXVIII) can be performed using an appropriate acid such as trifluoroacetic acid to obtain the compound of formula (XXXXIX).

[0089] Scheme 12 [ka] In the above scheme, B 1 is a halide or -SO2Me, R4 is a halogen or alkyl, A1 is -N- or -CH-, X is -O- or -NH, PG 2 and PG 6is, independently, a protecting group such as a tert-butoxycarbonyl group. In some cases, PG 6 has no protecting group.

[0090] The compound of formula (XXXXX) can be obtained by substituting the compound of formula (XXXIII) with the compound of formula (IX) in the presence of a suitable base such as DIEA.

[0091] Deprotection / overall deprotection of the compound of formula (XXXXX) can obtain the compound of formula (XXXXXI) using a suitable acid such as trifluoroacetic acid.

[0092] Therefore, the present invention also relates to a method for preparing the compound according to the present invention by the above procedure.

[0093] Pharmaceutical composition Another embodiment of the present invention provides a pharmaceutical composition or medicament containing the compound of the present invention and a therapeutically inert carrier, diluent or excipient, and a method of using the compound of the present invention for preparing such a composition and medicament. In one example, the compound of formula (I) may be formulated by mixing with a physiologically acceptable carrier, i.e., a carrier that is not toxic to the recipient at the dosages and concentrations used in galenical dosage forms, at a suitable pH and desired purity, at ambient temperature. The pH of the formulation mainly depends on the particular use and the concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula (I) is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula (I) is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0094] The composition is formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the dosing schedule, and other factors known to the medical practitioner.

[0095] The compounds of the present invention may be administered by any suitable means, including orally, topically (including buccal and sublingual), rectally, vaginally, percutaneously, parenterally, subcutaneously, intraperitoneally, intrapulmonaryly, intradermally, intrathecally, epidurally, and intranasally, and, if desired for topical treatment, intrafocal administration. Parenteral administration includes intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration.

[0096] The compounds of the present invention can be administered in any convenient dosage form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components that are conventional in pharmaceutical preparations, such as diluents, carriers, pH modifiers, sweeteners, fillers, and further activators.

[0097] Typical formulations are prepared by mixing the compound of the present invention with a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, Ansel, Howard C., et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C., Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow enhancers, processing aids, colorants, sweeteners, fragrances, flavorings, diluents, and other known additives for providing a drug (i.e., the compound of the present invention or its pharmaceutical composition) in an appealing manner, or for assisting in the manufacture of a pharmaceutical product (i.e., a drug).

[0098] The present invention will be better understood by referring to the following embodiments. However, they are not to be construed as limiting the scope of the invention. [Examples]

[0099] Abbreviation ACN: Acetonitrile; BH3·Me2S: Borane dimethyl sulfide complex; B℃: tert-butoxycarbonyl; CaCl2: Calcium chloride; Cbz: Carbobenzoxyl; CDI: Carbonyl diimidazole; DCE: 1,2-Dichloroethane; DCE: Dichloroethylene; DCM: Dichloromethane; DIEA: N,N-Diisopropylethylamine; DIPEA: N,N-Diisopropylethylamine; DMF: N,N-Dimethylformamide; DMSO: Dimethyl sulfoxide; DPPA: Diphenyl phosphoryl azide (Diphenyl phosphoroazidate); EA: Ethyl acetate; ESI: Electrospray ionization; Â: Ethyl acetate; Et3SiH: Triethylsilane; FA: Formic acid; H2O: Water; HATU: Azabenzotriazole tetramethyluronium hexafluorophosphate; HCl: Hydrochloric acid; HOsu: N-Hydroxylsuccinimide; HPLC: High-performance liquid chromatography; K2CO 3: Potassium carbonate; LCMS: Liquid chromatography-mass spectrometry; LiHMDS: Lithium bis(trimethylsilyl)amide; MeOH: Methyl alcohol; m-CPBA: Metachloroperbenzoic acid; MS: Mass spectrometry; MTBE: Methyl tert-butyl ether; NH4Cl: Ammonium chloride; n-Bu3P: Tributylphosphine; NaHCO3: Sodium bicarbonate; Na2SO4: Sodium sulfate; N2: Nitrogen; Na2SO3: Sodium sulfite; NH2OH·HCl: Hydroxylammonium chloride; NaH: Sodium hydride; NMP: N-methyl-2-pyrrolidinone; NMR: Nuclear magnetic resonance; NH3·H2O: Ammonium hydroxide; PdCl2: Palladium chloride; Pd / C: Palladium carbon; Pd(OH)2 / C: Carbon-supported palladium hydroxide; PE: Petroleum ether; Rf: Retention rate; SFC: Supercritical fluid chromatography; THF: Tetrahydrofuran; T3P: 2,4,6-tripropyl-1,3,5,2λ 5 ,4λ 5 ,6λ 5 -Trioxatriphosfinan 2,4,6 trioxide; Te℃: 2-(trimethylsilyl)ethoxycarbonyl; TEA: triethylamine; TFA: trifluoroacetic acid; TLC: thin-layer chromatography.

[0100] Intermediate and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and Quad12 / 25 Cartridge module, ii) ISCO combi-flash chromatography apparatus. Silica gel brands and pore sizes: i) KP-SIL 60Å, particle size: 40-60 μm; ii) CAS registry number: silica gel: 63231-67-4, particle size: 47-60 microns; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore size: 200-300 or 300-400.

[0101] Intermediates and final compounds were purified using XBridge® Prep-C18 (5 μm, OBD® 30 × 100 mm) columns, SunFire® Prep-C18 (5 μm, OBD® 30 × 100 mm) columns, Phenomenex Synergi-C18 (10 μm, 25 × 150 mm) or Phenomenex Gemini-C18 (10 μm, 25 × 150 mm). Waters AutoP purification system (Sample Manager 2767, Pump 2525, Detectors: Micromass ZQ and UV2487, Solvent system: Acetonitrile and 0.1% ammonium hydroxide aqueous solution; Acetonitrile and 0.1% FA aqueous solution or Acetonitrile and 0.1% TFA aqueous solution). Alternatively, the solution was purified by preparative HPLC on a reversed-phase column using a Gilson-281 purification system (pump 322, detector: UV156, solvent system: acetonitrile and 0.05% ammonium hydroxide aqueous solution; acetonitrile and 0.225% FA aqueous solution; acetonitrile and 0.05% HCl aqueous solution; acetonitrile and 0.075% TFA aqueous solution; or acetonitrile and water).

[0102] For SFC chiral separation, intermediates were separated using chiral columns (Daicel chiralpak IC, 5 μm, 30 × 250 mm), AS (10 μm, 30 × 250 mm), or AD (10 μm, 30 × 250 mm) with a Mettler Toledo Multigram III system SFC, Waters 80Q preparative SFC, or Thar 80 preparative SFC, solvent system: CO2 and IPA (0.5% TEA in IPA) or CO2 and MeOH (0.1% NH3·H2O in MeOH), back pressure of 100 bar, and detection UV at 254 or 220 nm.

[0103] The LC / MS spectra of the compounds were obtained using LC / MS (Waters® Alliance 2795-Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ, or Agilent Alliance 6110-Micromass ZQ), with the following LC / MS conditions (trial time 3 minutes or 1.5 minutes). Acidic conditions I: A: 0.1% TFA in H2O; B: 0.1% TFA in acetonitrile; Acidic conditions II: A: 0.0375% TFA in H2O; B: 0.01875% TFA in acetonitrile; Basic conditions I: A: 0.1% NH3·H2O in H2O; B: Acetonitrile; Basic conditions II: A: 0.025% NH3·H2O in H2O; B: Acetonitrile; Neutral conditions: A: H2O; B: Acetonitrile.

[0104] Mass spectra (MS): Generally, only ions showing the primary mass are reported, and unless otherwise stated, the cited mass ions are positive mass ions (MH). + That is the case.

[0105] NMR spectra were acquired using a Bruker Avance 400MHz.

[0106] The microwave-assisted reactions were carried out on a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under an argon or nitrogen atmosphere. Reagents were used as received from commercial suppliers without further purification, unless otherwise stated.

[0107] Int-1-1: 2,6-Dichloro-4-[1,4-dioxaspiro[4.5]decan-8-yl(difluoro)methyl]pyridine

Chem.

[0108] Step 1: Ethyl 1,4-dioxaspiro[4.5]decan-8-carboxylate

Chem.

[0109] Step 2: 1,4-Dioxaspiro[4.5]decan-8-carboxylic acid

Chem.

[0110] Step 3: N-Methoxy-N-methyl-1,4-dioxaspiro[4,5]decane-8-carboxamide [ka] To a solution of Int-1-1b (7.5 g, 40.3 mmol) in DMF (100 mL), HATU (23.0 g, 60.42 mmol) was added. Then, O,N-dimethylhydroxylamine·HCl (5.89 g, 60.42 mmol) and TEA (16.8 mL, 120.83 mmol) were added to the reaction mixture. This mixture was stirred at room temperature for 1 hour. After completion, the resulting mixture was diluted with EA (150 mL) and washed with water (100 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain the crude product, which was purified by silica gel column chromatography (PE with EA = 0%~10%) to obtain Int-1-1c (8 g, 34.89 mmol, yield 86.63%) as a colorless oil. MS measured value (ESI + ):230.1[(M+H) + ].

[0111] Step 4: (2,6-dichloro-4-pyridyl)-(1,4-dioxaspiro[4.5]decane-8-yl)methanone [ka] A solution of n-BuLi (16.57 mL, 39.78 mmol) in THF (50 mL) was mixed with a solution of 2,6-dichloropyridine (5.89 g, 39.78 mmol) in THF (20 mL) under N2 at -78°C. After mixing, the mixture was stirred for 30 minutes, and then a solution of Int-1-1c (7.6 g, 33.15 mmol) in THF (20 mL) was added. The mixture was then stirred at -78°C for 5 hours. After completion, the reaction mixture was quenched with an aqueous NH4Cl solution (100 mL) and extracted with EA (150 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The crude residue was purified by silica gel column (PE with EA = 0%~10%) to obtain Int-1-1d (6 g, 18.98 mmol, yield 57.25%) as a pale yellow solid. MS measured value (ESI + ):316.0[(M+H) + ].

[0112] Step 5: 2,6-Dichloro-4-[1,4-Dioxaspiro[4.5]decane-8-yl(difluoro)methyl]pyridine [ka] To a solution of Int-1-1d (4.0 g, 12.65 mmol) in DCM (20 mL), BAST (20 mL) was added. The mixture was stirred at 25°C for 3 hours. After completion, the mixture was quenched by slowly pouring it into cooling water (50 mL). The mixture was extracted using DCM (50 mL x 3). The combined organic layer was concentrated under vacuum, and the crude product was purified by silica gel column (PE with EA = 0%~10%) to obtain Int-1-1 (3.2 g, 9.46 mmol, yield 74.8%) as a white solid. MS measured value (ESI + ):338.0[(M+H) + ].

[0113] Int-2-1:4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexanone [ka] Compound Int-1-1 (500 mg, 1.48 mmol) was dissolved in MeOH (12 mL). 6 M HCl (250 μL, 1.48 mmol) was added to the suspension. The reaction mixture was then mixed for 90°C. ℃ The mixture was stirred overnight. After completion, the reaction mixture was cooled to room temperature and diluted with EA (30 mL) and water (30 mL). The aqueous layer was extracted with EA (20 mL x 2). The combined organic layers were washed with water (30 mL) and brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude residue. This was purified by silica gel column chromatography (EA in PE = 0%-50%) to obtain Int-2-1 (300 mg, 68.99%) as a pale yellow solid. MS measured value (ESI + )[(M+H) + ]:294.232.

[0114] Int-3-1:4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexaneamine [ka] To a solution of Int-2 (1.74 g, 5.92 mmol) in MeOH (30 mL), ammonium acetate (4.56 g, 59.16 mmol) and sodium triacetoxyborohydride (2.51 g, 11.83 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. After completion, the mixture was poured into water (50 mL) at 0°C. The mixture was extracted with EA (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain a crude residue. This was purified by silica gel column chromatography (MeOH in DCM = 0%~30%) to obtain Int-3-1 (1.6 g, 5.42 mmol, yield 91.63%) as a colorless oil. MS measured value (ESI + ):295.0[(M+H) + ].

[0115] Int-4-1:Cis-4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexaneamine [ka] Int-4-1 was prepared using cis-4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexanone instead of Int-2-1, in the same manner as the preparation of compound Int-3-1. Int-4-1 (200 mg) was obtained as a white solid. MS measurement values ​​(ESI) + )[(M+H) + ]:296.6.

[0116] Int-1:tert-butylN-[(3R)-5-oxo-1-(4-piperazine-1-ylsulfonylphenyl)pyrrolidine-3-yl]carbamate [ka]

[0117] Step 1: Benzyl 4-(4-nitrophenyl)sulfonylpiperazine-1-carboxylate [ka] To a solution of 1-Cbz-piperazine (49.7 g, 225.6 mmol) and TEA (94.3 mL, 676.8 mmol) in DCM (500 mL), 4-nitrobenzenesulfonyl chloride (50.0 g, 225.6 mmol) was added in an ice bath at 0°C. After stirring at 0°C for 2 hours, H2O (500 mL) was added to the mixture, and it was extracted with DCM (500 mL x 3). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with MTBE to obtain Int-1a (87.0 g, 214.6 mmol) as a white solid. MS measured value (ESI + )[(M+H) + ]:428.1.

[0118] Step 2: Benzyl 4-(4-aminophenyl)sulfonylpiperazine-1-carboxylate [ka] To a solid of compound Int-1a (87.0 g, 214.6 mmol) and Pt / C (4.0 g), isopropanol (400 mL) and THF (400 mL) were added under N2 at 25°C. The reaction mixture was purged three times with H2 and stirred under H2 (45 Psi) at 45°C for 72 hours. After cooling to room temperature, degassing under vacuum, and purging several times with N2, the reaction mixture was diluted with THF (1.0 L), filtered through a Celite pad, and washed with THF (1.0 L). The filtrate was concentrated under vacuum to obtain Int-1b (81.0 g, 215.8 mmol) as a white solid. MS measurement value (ESI+)[(M+H)+]: 376.1.

[0119] Step 3: Benzyl 4-[4-[[(3R)-3-(tert-butoxycarbonylamino)-4-hydroxy-butanoyl]amino]phenyl]sulfonylpiperazine-1-carboxylate [ka] A solution of compound Int-1b (50.0 g, 133.2 mmol) in THF (666 mL) was added dropwise at 0°C to a solution of LiHMDS in THF (1.0 M, 333 mL, 333 mmol), and the mixture was stirred at 0°C for 0.5 hours. Next, tert-butyl N-[(3R)-5-oxotetrahydrofuran-3-yl]carbamate (40.2 g, 199.77 mmol) was added at 0°C, and the mixture was stirred at 0°C for 1 hour. The reaction mixture was quenched by adding water (500 mL) and extracted with ethyl acetate (500 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent. The residue was collected by silica gel column (PE / ethyl acetate = 5 / 1~1 / 3; TLC: PE / ethyl acetate = 1 / 1, R f Purified by (=0.3), Int-1c (55.0 g, 95.4 mmol, yield 71.6%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:577.3.

[0120] Step 4: Benzyl 4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-carboxylate [ka] To a solution of Bu3P (32.5 mL, 130.1 mmol) and compound Int-1c (50.0 g, 86.7 mmol) in THF (500 mL), N,N,N,N-tetramethylazodicarboxamide (CAS: 10465-78-8, 22.4 g, 130.1 mmol) was added in fractions under N2 at 0°C. The mixture was then heated to 25°C and stirred for 2 hours. After adding aqueous HCl (1 M, 200 mL), the mixture was extracted with RINKAN (500 mL x 3). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent. The residue was triturated with MTBE (500 mL) to obtain Int-1d (45.0 g, 80.6 mmol, yield 92.9%) as a white solid. MS measured values ​​(ESI + )[(M+H) + ]:559.2.

[0121] Step 5: tert-butyl N-[(3R)-5-oxo-1-(4-piperazine-1-ylsulfonylphenyl)pyrrolidine-3-yl]carbamate [ka] To a solution of compound Int-1d (45.0 g, 80.6 mmol) in isopropanol (250 mL) and THF (250 mL), wet Pd / C (5.0 g, 50% water content) was added under N2 at 25°C. The reaction mixture was degassed under vacuum and purged three times with H2. The suspension was then stirred under H2 (45 Psi) at 45°C for 12 hours. The reaction mixture was cooled to room temperature, purged with N2, diluted with THF (1.0 L), filtered through a Celite pad, and washed with a solution of THF (2.0 L) and MeOH (2.0 L). The filtrate was concentrated under vacuum to obtain crude Int-1 (38.0 g, 89.5 mmol, yield 94.5%) as a brown solid. The crude product was used directly in the next step without further purification. MS measured value (ESI+)([M+H]) + ):425.2.

[0122] Int-2:Tert-butylN-[rac-(3R)-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Step 1: tert-butyl N-(4-formylcyclohexyl)carbamate [ka] To a solution of oxalyl chloride (41.5 g, 327.1 mmol) in DCM (200 mL), a solution of DMSO (37.5 g, 479.7 mmol) in DCM (100 mL) was added dropwise under N2 at -78°C, and the mixture was stirred under N2 at -78°C for 1 hour. Next, a solution of tert-butyl N-[4-(hydroxymethyl)cyclohexyl]carbamate (50.0 g, 218.0 mmol) in DCM (100 mL) was added at -78°C, and the mixture was stirred for 1 hour. After adding DIEA (112.9 mL, 872.1 mmol) to the mixture at -78°C, the reaction mixture was heated to 20°C over 1 hour, and stirred continuously at 20°C for 1 hour. The reaction mixture was quenched with water (300 mL) at room temperature and extracted with DCM (300 mL x 3). The combined organic layers were washed with HCl aqueous solution (1.0 M, 100 mL x 3) and saturated brine (200 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain crude Int-2a (48.0 g, 211.2 mmol, yield 96.9%) as a pale yellow solid. The crude product was used directly in the next step without further purification.

[0123] Step 2: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-hydroxymethyl]cyclohexyl]carbamate [ka] To a mixture of 2,6-dichloro-4-iodopyridine (86.8 g, 316.8 mmol) (recrystallized with toluene at 110°C) in THF (200 mL), isopropyl magnesium chloride-lithium chloride complex (268.0 mL, 348.4 mmol) was added dropwise under N2 at -70 to -40°C. The mixture was stirred at 20°C for 2 hours. A solution of Int-2a (48.0 g, 211.2 mmol) in THF (200 mL) was added to the mixture at -70 to -40°C, and the reaction mixture was heated to 0°C and stirred at 0°C for 0.5 hours. The reaction mixture was poured into saturated NH4Cl aqueous solution (300 mL) and extracted with ELISA (300 mL x 4). The combined organic layers were washed with saturated brine (300 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude product obtained was triturated twice with SiO(150 mL) to obtain compound Int-2b (22.0 g, 58.6 mmol, yield 27.8%) as a white solid. MS measured value (ESI+)([{ 35 Cl}M-C4H8+H] + ):319.0.

[0124] Step 3: Tert-butyl N-[4-(2,6-dichloropyridine-4-carbonyl)cyclohexyl]carbamate [ka] To a solution of compound Int-2b (22.0 g, 58.6 mmol) in THF (400 mL), DMP (49.7 g, 117.2 mmol) was added in fractions at 0°C. After the addition of DMP, the mixture was heated to 20°C and stirred for 4 hours. The reaction mixture was poured into saturated K2CO3 aqueous solution (600 mL) and the mixture was stirred at 20°C for 12 hours. The mixture was extracted with siRNA (400 mL x 3). The combined organic layer was washed with saturated brine (150 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain Int-2c (17.0 g, 45.5 mmol, yield 77.7%) as a pale yellow solid. MS measured value (ESI+)([{ 35 Cl}M-C4H8+H] + ):316.9.

[0125] Step 4: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]carbamate [ka] To a solution of compound Int-2c (17.0 g, 45.5 mmol) in DCM (60 mL), DAST (34.0 mL) was added at 0°C. The reaction mixture was stirred at 20°C for 16 hours, then slowly poured into ice water (400 mL), and the mixture was extracted with DCM (200 mL x 3). The combined organic layers were washed with saturated NaHCO3 aqueous solution (200 mL) and saturated brine (100 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was collected on a silica gel column (PE / siRNA=20 / 1~5 / 1; TLC:PE / siRNA=2 / 1, R f Purified by (=0.5), Int-2d (11.6g, 29.4 mmol, yield 64.4%) was obtained as a pale yellow solid. MS measured value (ESI+)([{ 35 Cl}M-C4H9+H] + ):339.1.

[0126] Step 5: 4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexaneamine [ka] To a solution of compound Int-2d (5.0 g, 12.7 mmol) in DCM (10 mL), TFA (10.0 mL) was added at 25°C. After stirring at 25°C for 1 hour, the reaction mixture was concentrated under vacuum to obtain crude Int-2e (3.5 g, 11.9 mmol, yield 93.7%) as a dark green oil. The crude product was used directly in the next step without purification. MS measured value (ESI+)([{ 35 Cl}M+H] + ):294.8.

[0127] Step 6: 2-(trimethyl-λ4-sulfanyl)ethyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]carbamate [Chemical formula] To a solution of compound Int-2e (3.5 g, 11.9 mmol) and DIEA (6.2 mL, 35.6 mmol) in DCM (50 mL) was added Te℃-OSu (3.7 g, 14.2 mmol) at 0 °C. Then, it was stirred at 25 °C for 2 hours. The reaction mixture was diluted with H2O (100 ml), extracted with DCM (100 mL × 3), and washed with saturated brine (100 ml). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by silica gel column (PE / EtOAc = 1 / 0 to 5 / 1; TLC: PE / EtOAc = 2 / 1, R f = 0.45) to obtain Int-2f (3.6 g, 8.1 mmol, 85.9% yield) as a yellow oil. Measured MS value (ESI+) ([{ 35 Cl}M-C2H4+H] + ): 411.2.

[0128] Step 7: N-[4-[[2-Chloro-6-[4-[4-[rac-(4R)-4-(tert-Butoxycarbonylamino)-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4-pyridyl]-difluoromethyl]cyclohexyl]-2-trimethylsilylethyl carbamate [Chemical formula] To a solution of compound Int-2f (700.0 mg, 1.6 mmol) and Int-1 (743.9 mg, 1.8 mmol) in DMSO (4 mL) was added DIEA (4.7 mL, 26.8 mmol) at once. After stirring at 100 °C for 12 hours, the reaction mixture was cooled to 25 °C and poured into H2O (50 mL). The mixture was extracted with EtOAc (150 mL × 2) and washed with saturated brine (50 mL × 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to obtain Int-2g (1.5 g, 1.8 mmol, 97.9% yield) as a yellow oil. The crude product was used directly in the next step without further purification. Measured MS value (ESI+) ([{35 Cl}M + H] + ): 827.5, ([{ 35 Cl}M - C2H4 + H] + ): 799.5, ([{ 35 Cl}M - B℃ - C2H4 + H] + ): 699.5.

[0129] Project 8: Tert-butyl N-[rac-(3R)-1-[4-[4-[4-[(4-Aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate [[ID=​​​​​​​​​​​​​​​​​​​In step 1, Int-3 was prepared in the same manner as in the preparation of compound Int-2d by using cis-tert-butyl N-[4-(hydroxymethyl)cyclohexyl]carbamate instead of trans-tert-butyl N-[4-(hydroxymethyl)cyclohexyl]carbamate. Int-3 (200 mg) was obtained as a white solid. MS measured value (ESI+)([{ 35 Cl}M-C4H9+H] + ):339.1,([{ 35 Cl}MB℃+H] + ):294.8.

[0131] Int-4:Tert-butylN-[rac-(3R)-1-[4-[4-[4-[[4-(aminomethyl)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka]

[0132] Step 1: Methyl 4-(hydroxymethyl)cyclohexanecarboxylate [ka] To a solution of 4-(methoxycarbonyl)cyclohexane-1-carboxylic acid (50.0 g, 268.5 mmol) in THF (150 mL), BH3·Me2S (10 M, 54 mL, 540 mmol) was added dropwise under N2 at 0-10°C for 1 hour. The mixture was stirred at 25°C for 12 hours. After cooling the reaction mixture to 0°C, the reaction was quenched by adding MeOH (100 mL) dropwise to the reaction mixture at 0°C. The mixture was concentrated under vacuum and dried at 40°C. The residue was filtered through a silica gel column (PE / Depositphotos = 100 / 1-1 / 1; TLC: PE / Depositphotos = 2 / 1, R f Purified by (=0.4), Int-4a (28.0 g, 162.6 mmol, yield 60.6%) was obtained as a colorless oil. Int-4a was used directly in the next step without analysis. TLC of commercially available standard samples was performed.f This was determined by comparing the two. 1 H NMR(400MHz,DMSO-d6)δ ppm3.58(s,3H),3.19-3.22(d,J=4Hz,2H),2.22-2.23(m,1H),1.77-1.92(m,2H),1.73-1.75 (m,2H),1.27-1.32(m,3H),0.91-0.96(td,J=9.6,3.6Hz,2H).MS actual value (ESI+)[(M+H)+]:173.2.

[0133] Step 2: Methyl-4-formylcyclohexanecarboxylate [ka] To a solution of oxalyl chloride (31.0 g, 243.9 mmol) in DCM (100 mL), DMSO (25.4 g, 325.2 mmol) was added at -60°C, and the mixture was stirred under N2 at -60°C for 0.5 hours. After adding a solution of compound Int-4a (28.0 g, 162.6 mmol) in DCM (100 mL) at -60°C, the reaction mixture was stirred at -60°C for 1 hour. Next, DIEA (84.1 g, 650.3 mmol) was added at -60°C, and the mixture was continuously stirred at -60 to -20°C for 1 hour. Then, the mixture was heated to 0°C and stirred for a further 1 hour under N2. The mixture was diluted with siRNA (600 mL), washed with aqueous CaCl2 solution (100 mL x 3) and saturated brine (100 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. Residue in silica gel column (PE / Â=30 / 1~1 / 1; TLC:PE / Â=3 / 1, R f Purified by (=0.5), Int-4b (24.0 g, 141.0 mmol, yield 86.7%) was obtained as a pale yellow oily substance. Int-4b was used directly in the next step without characterization analysis. TLC of commercially available standard samples f This was determined by comparing the two. 1 H NMR(400MHz,DMSO-d6)δ ppm9.56-9.58(s,1H),2.24-2.29(m,2H),1.93-2.00(m,4H),1.26-1.34(m,4H)

[0134] Step 3: 4-[(2,6-dichloro-4-pyridyl)-hydroxymethyl]cyclohexanecarboxylate [ka] A solution of 2,6-dichloro-4-iodopyridine (42.5 g, 155.1 mmol) in THF (50 ml) was mixed dropwise with a solution of isopropylmagnesium chloride lithium chloride complex (130.2 mL, 169.2 mmol) in THF under N2 at -40°C. The mixture was stirred at 25°C for 1 hour. Then, the mixture was mixed dropwise with a solution of compound Int-4b (24.0 g, 141.0 mmol) in THF (100 mL) at -40°C. The reaction mixture was stirred at 25°C for 3 hours. The mixture was poured into an aqueous solution of NH4Cl (200 mL) and extracted with ethyl acetate (300 mL x 3). The combined organic layers were washed with saturated brine (100 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was filtered through a silica gel column (PE / ethyl acetate = 100 / 1~1 / 2; TLC: PE / ethyl acetate = 1 / 1, R f The compound Int-4c (33.0 g, 103.7 mmol, yield 73.6%) was purified by (=0.4) and obtained as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:317.8.

[0135] Step 4: Methyl 4-(2,6-dichloropyridine-4-carbonyl)cyclohexanecarboxylate [ka] To a solution of compound Int-4c (33.0 g, 103.7 mmol) in THF (150 mL), desmartin periodinane (48.4 g, 114.1 mmol) was slowly added under N2 at 0°C. The mixture was then heated to 25°C and stirred under N2 for 3 hours. The mixture was diluted with ELISA (500 mL) and poured into K2CO3 aqueous solution (200 mL). The mixture was stirred at room temperature for 0.5 hours and filtered. The organic layer was separated, and the combined organic layers were washed with K2CO3 aqueous solution (100 mL x 2) and saturated brine (100 mL x 2). The mixture was dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain the crude compound Int-4d (30.0 g, 94.9 mmol, yield 91.5%) as a white solid. The crude product was used directly in the next step without further purification. MS measurement values ​​(ESI) + )[({ 35 Cl}M+H) + ]:315.8.

[0136] Step 5: Methyl 4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexanecarboxylate [ka] A solution of compound Int-4d (29.5 g, 93.3 mmol) in DCM (20 mL) was mixed with diethylaminosulfur trifluoride (135.4 g, 839.7 mmol) and stirred at 45°C for 12 hours. After cooling to 25°C, the mixture was poured into ice water at 0°C and then extracted with siRNA (200 mL x 3). The combined organic layer was washed with aqueous K2CO3 solution (100 mL x 2) and saturated brine (100 mL x 2), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum. The residue was recrystallized with MeOH (150 mL) to obtain compound Int-4e (25.6 g, 75.7 mmol, yield 81.1%) as a white solid. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:337.8.

[0137] Step 6: [4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]methanol [ka] To a solution of compound Int-4e (250.0 mg, 0.7 mmol) in THF (5 mL), LiAlH4 (42.1 mg, 1.1 mmol) was added at -10°C, and the mixture was stirred at -10°C for 2 hours. After warming the mixture to 0°C, the reaction mixture was diluted with THF (100 mL) and quenched with H2O (0.1 mL), followed by 15% NaOH aqueous solution (0.2 mL) and H2O (0.1 mL). MgSO4 (10 g) was added to the suspension, and the mixture was stirred at 20°C for 0.5 hours. The mixture was filtered through a Celite pad, and the filtrate was concentrated under vacuum. The residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain Int-4f (210.0 mg, 0.7 mmol, yield 91.6%) as a white solid. MS measured values ​​(ESI) + )[({ 35 Cl}M+H) + ]:310.1

[0138] Step 7: Tert-butyl N-[rac-(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(hydroxymethyl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of compound Int-4f (210.0 mg, 0.7 mmol) in DMSO (1 mL), Int-1 (288.1 mg, 0.7 mmol) was added at 20°C, and the mixture was stirred at 120°C for 16 hours. After cooling to room temperature, the resulting mixture was transferred to a separatory funnel, and the aqueous layer was extracted with SiO2 (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. This crude product was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and Int-4 g (400.0 mg, 0.6 mmol, yield 77.4%) was obtained as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]::698.2.

[0139] Step 7: Tert-butyl N-[rac-(3R)-1-[4-[4-[4-[[4-(aminomethyl)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of compound Int-4g (380.0mg, 0.5 mmol) in DCM (5 mL), trifluoromethanesulfonic anhydride (0.2 mL, 1.1 mmol) was added at -20°C, and the mixture was stirred at -20°C for 0.5 hours. The mixture was then poured into ammonia THF solution (20 mL) and stirred at 45°C for 1 hour. After concentrating the mixture to 10 mL, it was diluted by slowly adding saturated ammonium chloride aqueous solution (20 mL). The resulting mixture was transferred to a separatory funnel, and the aqueous layer was extracted with  (50 mL × 3). The combined organic layers were washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain Int-4 (120.0 mg, 0.2 mmol, yield 31.6%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:697.2.

[0140] Int-5:tert-butyl((R)-1-(4-((4-(((1r,4r)-4-aminocyclohexyl)difluoromethyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka]

[0141] Step 1: Chlorolithium; Chloro-(2,2,6,6-tetramethyl-1-piperidyl)magnesium [ka] To a solution of isopropylmagnesium chloride-lithium chloride complex (50.0 mL, 65.0 mmol), 2,2,6,6-tetramethylpiperidine (12.1 mL, 71.5 mmol) was added dropwise at -40°C. The mixture was stirred under N22 at 25°C for 24 hours to obtain a THF solution of Int-5a (60 mL, approximately 1.1 M). The solution was used directly in the next step without further manipulation.

[0142] Step 2: Methyl 4-[(6-chloro-2-methylsulfanylpyrimidine-4-yl)-hydroxymethyl]cyclohexanecarboxylate [ka] To a solution of 4-chloro-2-(methylthio)pyrimidine (3.0 g, 18.7 mmol) in THF (8 mL), Int-5a (18.7 mL, 20.6 mmol) was added at -40°C and stirred at 0°C for 40 minutes. The mixture was added to a solution of methyl 4-formylcyclohexanecarboxylate (3.5 g, 20.6 mmol) in THF (12 mL) at -40°C, then heated to 25°C and stirred for 1 hour. The mixture was poured into water (50 mL) and extracted with ethyl acetate (80 mL x 3). The organic layer was washed with saturated brine (50 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was filtered through a silica gel column (PE / ethyl acetate = 100 / 1~1 / 1; TLC: PE / ethyl acetate = 1 / 1, R f The sample was purified by (=0.4) to obtain Int-5b (1.8g, 5.4 mmol, yield 29.1%) as a pale yellow oily substance. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:331.3

[0143] Step 3: Methyl 4-(6-chloro-2-methylsulfanylpyrimidine-4-carbonyl)cyclohexanecarboxylate [ka] To a solution of compound Int-5b (1.7 g, 5.1 mmol) in THF (30 mL), desmartin periodinane (2.6 g, 6.2 mmol) was added at 0°C, and the mixture was stirred at 25°C for 2 hours. The mixture was quenched with ice water (30 mL) and adjusted to pH=9 with NaHCO3 aqueous solution. The mixture was extracted with ethyl acetate (100 mL x 2) and washed with saturated brine (50 mL x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was filtered through a silica gel column (PE / ethyl acetate = 1 / 0 to 1 / 1; TLC: PE / ethyl acetate = 1 / 1, R f Purified by (=0.5), Int-5c (1.6g, 4.9 mmol, yield 94.7%) was obtained as a yellow solid. (ESI + )[({ 35 Cl}M+H) + ]:329.3.

[0144] Step 4: Methyl 4-[(6-chloro-2-methylsulfanylpyrimidine-4-yl)-difluoromethyl]cyclohexanecarboxylate [ka] To a solution of Int-5c (1.6 g, 4.9 mmol) in DCM (10 mL), diethylaminosulfur trifluoride (10.0 mL, 4.9 mmol) was added at 0°C, and the mixture was heated to 25°C and stirred for 2 hours. The mixture was carefully poured into ice water and adjusted to pH=9 with NaHCO3 aqueous solution. The mixture was extracted with ethylacetate (200 mL x 2) and washed with saturated brine (100 mL x 2). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was filtered through a silica gel column (PE / ethylacetate=1 / 0~5 / 1; TLC:PE / ethylacetate=5 / 1, R f Purified by (=0.55), Int-5d (1.5g, 4.3 mmol, yield 87.9%) was obtained as a yellow solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:351.3.

[0145] Step 5: Methyl 4-[difluoro-(6-methyl-2-methylsulfanylpyrimidine-4-yl)methyl]cyclohexanecarboxylate [ka] Trimethylboroxine (9.7 mL, 30.8 mmol) was added at 25°C to a mixture of compound Int-5d (900.0 mg, 2.6 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (201.9 mg, 0.3 mmol), and potassium phosphate (1.6 g, 7.7 mmol) in 1,4-dioxane (5 mL) and water (2 mL). The reaction mixture was degassed under vacuum, purged several times with N2, and stirred at 80°C for 12 hours under N2. After cooling to room temperature, the mixture was poured into H2O (20 mL), extracted with ELISA (50 mL x 2), and washed with brine (50 mL x 2). The combined organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was placed in a silica gel column (PE / Â=30 / 1~10 / 1; TLC:PE / Â=5 / 1, R f The sample was purified by (=0.65) and Int-5e (420.0 mg, 1.3 mmol, yield 34.7%) was obtained as a yellow oily substance. MS measured value (ESI + )[(M+H) + ]:331.3.

[0146] Step 6: (1r,4r)-4-(difluoro(6-methyl-2-(methylthio)pyrimidine-4-yl)methyl)cyclohexane-1-carboxylic acid: [ka] To a mixture of Int-5e (2.3 g, 7.0 mmol) in MeCN (10 mL) and water (2 mL), LiBr (6.0 g, 70.0 mmol) and TEA (2.1 g, 21.0 mmol) were added at 25°C. After stirring the reaction mixture at 50°C for 2 hours, the mixture was quenched by slowly adding HCl solution (1.0 M, 20 mL). The resulting mixture was transferred to a separatory funnel, and the aqueous layer was extracted with SiO2 (100 mL x 3). The combined organic layers were washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product Int-5f (2.0 g, 6.3 mmol, yield 90.3%) as a white solid, which was used in the next step without further purification. MS measured values ​​(ESI + )[(M+H) + ]:317.1.

[0147] Step 7: 4-(difluoro((1r,4r)-4-isocyanatocyclohexyl)methyl)-6-methyl-2-(methylthio)pyrimidine [ka] To a solution of Int-5f (2.0 g, 6.3 mmol) in THF (10 mL), DPPA (3.7 g, 13.3 mmol) and TEA (2.2 g, 22.1 mmol) were added at 25°C. After stirring the reaction mixture at 25°C for 2 hours, the mixture was quenched by slowly adding saturated ammonium chloride aqueous solution (20 mL). The resulting mixture was transferred to a separatory funnel, and the aqueous layer was extracted with Depositphotos (100 mL x 3). The combined organic layers were washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product, which was then analyzed on a silica gel column (PE / Depositphotos=1 / 0~2 / 1; TLC:PE / Depositphotos=2 / 1, R f Purified by (=0.5), Int-5g (1.0g, 3.2 mmol, yield 50.5%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:314.0

[0148] Step 8: Benzyl((1r,4r)-4-(difluoro(6-methyl-2-(methylthio)pyrimidine-4-yl)methyl)cyclohexyl)carbamate [ka] Benzyl alcohol (20.0 mL) was added to Int-5 g (1.0 g, 3.2 mmol) at 25°C, and the mixture was heated to 50°C and stirred for 2 hours. After cooling to 25°C, the resulting mixture was poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with ethyl acetate (80 mL × 3). The combined organic layers were washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product, which was then analyzed on a silica gel column (PE / ethyl acetate = 1 / 0~2 / 1; TLC: PE / ethyl acetate = 1 / 1, R₂). f Purified by (=0.3), Int-5h (1.1g, 2.6 mmol, yield 81.8%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:422.1.

[0149] Step 9: Tert-butyl((1r,4r)-4-(difluoro(6-methyl-2-(methylsulfonyl)pyrimidine-4-yl)methyl)cyclohexyl)carbamate [ka] To a solution of Int-5h (1.1 g, 2.6 mmol) in acetic acid (20 mL), sodium tungstate dihydrate (172.2 mg, 0.5 mmol) and hydrogen peroxide (4.0 mL, 30% wt.) were added all at once under N2 at 0°C. The reaction mixture was stirred at 25°C for 16 hours, then quenched with aqueous NaHCO3 (100 mL) and aqueous Na2SO3 (100 mL), and subsequently extracted with ethyl acetate (100 mL x 3). The combined organic layer was washed with saturated brine (100 mL), dried with anhydrous Na2SO4, and then filtered. The filtrate was concentrated under vacuum to obtain the crude product, which was then filtered through a silica gel column (PE / ethyl acetate = 1 / 0 to 1 / 1; TLC: PE / ethyl acetate = 1 / 1, R). fPurification was performed using (=0.5) to obtain Int-5i (1.0 g, 2.2 mmol, yield 84.7%) as a white solid. MS measured value (ESI + )[(M+H) + ]:454.1.

[0150] Step 10: Tert-butyl((R)-1-(4-((4-(((1r,4r)-4-(((benzyloxy)carbonyl)amino)cyclohexyl)difluoromethyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] To a solution of Int-5i (1.0 g, 2.2 mmol) in DMSO (4 mL), DIEA (0.9 g, 6.6 mmol) and Int-1 (1.0 g, 2.4 mmol) were added. The reaction mixture was stirred at 100 °C for 16 hours and then cooled to room temperature. The resulting mixture was poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with SiO (80 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was filtered through a silica gel column (PE / SiO=1 / 0~0 / 1; TLC:PE / SiO=1 / 1, R f Purified by (=0.4), Int-5j (1.2g, 1.5 mmol, yield 67.9%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:798.6.

[0151] Step 11: Tert-butyl((R)-1-(4-((4-(((1r,4r)-4-aminocyclohexyl)difluoromethyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] To a solution of Int-5j (1.2 g, 1.5 mmol) in DCM (20 mL), Et3SiH (1.9 mL, 12.0 mmol), PdCl2 (132.4 mg, 0.8 mmol), and TEA (0.6 mL, 4.5 mmol) were added. The mixture was degassed under vacuum and purged three times with N2. The reaction mixture was stirred under N2 at 25°C for 2 hours, and the resulting mixture was poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with Depositphotos (100 mL × 3). The combined organic layers were washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was then filtered through a silica gel column ( Depositphotos / MeOH = 10:0~5:1; TLC: Depositphotos / MeOH = 5 / 1, R f Purified by (=0.5), Int-5 (870.0 mg, 1.3 mmol, yield 87.2%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:664.3.

[0152] Int-6:Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(hydroxyamino)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka]

[0153] Step 1: Benzyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]amino]propyl]carbamate [ka] To a solution of Int-2 (0.3 g, 0.4 mmol) in DMF (2 mL), bromoacetonitrile (105.3 mg, 0.9 mmol) and K2CO3 (181.8 mg, 1.3 mmol) were added. The reaction mixture was stirred at 20°C for 16 hours, then diluted with RINKAN (50 mL) and filtered. The filtrate was concentrated under vacuum. The residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain Int-6a (150.0 mg, 0.2 mmol, yield 47.3%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:722.3.

[0154] Step 2: N-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-1-cyano-methanimine oxide [ka] To a solution of compound Int-6a (200.0 mg, 0.3 mmol) in DCM (3 mL), m-CPBA (105.1 mg, 0.6 mmol) was added at 0°C. After stirring at 20°C for 2 hours, the reaction mixture was quenched with saturated Na2SO3 aqueous solution (10 mL), and the reaction mixture was extracted with DCM (70 mL x 3). The combined organic layers were successively washed with NaHCO3 (50 mL) and saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and Int-6b (100.0 mg, 0.1 mmol, yield 49.1%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:736.3.

[0155] Step 3: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(hydroxyamino)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of compound Int-6b (60.0 mg, 0.1 mmol) in methanol (1.0 mL) and DMF (1 mL), NH2OH·HCl (34.0 mg, 0.5 mmol) was added at 20°C, and the mixture was stirred at 60°C for 2 hours. The solvent was concentrated under vacuum, and the residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and Int-6 (50.0 mg, 0.1 mmol, yield 87.8%) as a white solid. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:699.3.

[0156] Int-7:Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(methylamino)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka]

[0157] Step 1: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]-N-methyl-carbamate [ka] To a solution of Int-2d (0.8 g, 2.0 mmol) and iodomethane (316.0 mg, 2.2 mmol) in DMF (2 mL), NaH (89.1 mg, 2.2 mmol) was added at 0°C and the mixture was stirred at 0°C for 1 hour. After quenching the reaction by adding ice water (10 mL), the mixture was extracted with RINKAN (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and then filtered. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain Int-7a (541.0 mg, 1.3 mmol, yield 65.3%) as a yellow oily substance. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:353.2.

[0158] Step 2: 4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]-N-methylcyclohexaneamine [ka] To a solution of Int-7a (500.0 mg, 1.2 mmol) in DCM (1 mL), TFA (1.0 mL, 13.0 mmol) was added at 25°C, and the mixture was stirred at this temperature for 1 hour. The mixture was concentrated under vacuum, and the residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and int.7b (320.0 mg, 1.0 mmol, yield 84.7%) was obtained as a yellow oily substance. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:309.0.

[0159] Step 3: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(methylamino)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of Int-7b (150.0 mg, 0.49 mmol) and Int-1 (206.0 mg, 0.5 mmol) in DMSO (1 mL), DIEA (190.0 mg, 1.5 mmol) was added and the mixture was stirred at 100°C for 16 hours. After cooling to 25°C, the mixture was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and Int-7 (240.0 mg, 0.3 mmol, yield 71.0%) was obtained as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:697.4.

[0160] Example 001: cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidine-2-one (001A) and trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidine-2-one (001B) [ka] [ka] Step 1: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[1,4-dioxaspiro[4.5]decane-8-yl(difluoro)methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compound Int-1-1 (200 mg, 591 μmol) and tert-butyl(R)-(5-oxo-1-(4-(piperazine-1-ylsulfonyl)phenyl)pyrrolidine-3-yl)carbamate (213 mg, 502 μmol) were dissolved in DMF (6 mL), and then DIEA (263 μl, 1.51 mmol) was added. The reaction mixture was stirred overnight at 120 °C. After the reaction mixture cooled to room temperature, it was diluted with EA (30 mL) and quenched with water (30 mL). The aqueous layer was extracted with EA (20 mL x 2). The combined organic layers were washed with water (30 mL) and brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain the crude residue. This was purified by silica gel column chromatography (PE with EA = 0%-70%) to obtain 001b (0.29 g, 399 μmol, yield 79.6%). MS measured value (ESI + ):726.201[(M+H) + ].

[0161] Step 7: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-(4-oxocyclohexyl)methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compound 001a (227 mg, 313 μmol) was dissolved in 36% HCl (5 mL, 25 mmol) and MeOH (5 mL). The mixture was stirred at 50°C for 2 hours. LC-MS mainly showed the formed product, but de-B°C products were also observed. The mixture was then adjusted to pH 8 with NaHCO3 aqueous solution and extracted with DCM (30 mL x 3). The combined organic layer was concentrated under vacuum to obtain the crude product (180 mg, 201 μmol, yield 64.3%) as a pale yellow solid. MS measured values ​​(ESI + ):582.375[(M+H) + The obtained compound was dissolved in DCM (2.5 mL). B℃2O (93.7 mg, 430 μmol) and Et3N (71.8 μl, 515 μmol) were added to the stirred solution. The reaction mixture was stirred overnight at room temperature, and LC-MS showed that the starting materials had been consumed. After completion, the reaction mixture was extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain crude 001b (100 mg, 147 μmol, yield 85.3%) as a yellow oily substance. MS measured value (ESI + ):682.152[(M+H) + ].

[0162] Step 8: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compound 001b (100 mg, 172 μmol) and azetidine-3-ol hydrochloride (94.1 mg, 859 μmol) were dissolved in THF (15 mL), and one drop of titanium(IV) isopropoxide was added. The reaction mixture was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (182 mg, 859 μmol) and 5 mL of MeOH were added. The mixture was heated to 70°C. ℃The mixture was stirred for 2 hours. After completion, the mixture was poured into water (50 mL) and extracted with EA (50 mL x 3). The combined organic layer was washed with brine (50 mL x 2), dried with anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain crude O01c (127 mg, 172 μmol, yield 100%), which was used directly in the next step. MS measured value (ESI + ):739.3[(M+H) + ].

[0163] Step 9: cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidine-2-one (001A) and trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidine-2-one (001B) [ka] [ka] Compound 001c (127 mg, 172 μmol) and TFA (23.1 mg, 203 μmol) were mixed in DCM (5 mL). The reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was concentrated under vacuum to obtain the crude compound, which was purified by preparative HPLC to obtain a mixture of 001A and 001B. The two epimers were separated by SFC to obtain Example 001A (first elution, 93 mg, yield 64%) and Example 001B (second elution, 14 mg, yield 20%). The cis and trans configurations were confirmed by 2D NMR.

[0164] Example 001A: MS measured value (ESI + )[(M+H) + ]:639.3. 11H NMR (400MHz, DMSO-d6)δ ppm7.88-7.98(m,2H),7.76-7.85(m,2H),6.71-6.81(m,1H),6.64-6.69(m,1 H),4.36-4.51(m,1H),4.18-4.36(m,3H),4.04-4.15(m,1H),3.74-3.95(m,3H ),3.59-3.73(m,4H),3.02-3.15(m,1H),2.88-3.02(m,4H),2.54-2.66(m,1H) ,2.17-2.34(m,1H),1.67-1.82(m,2H),1.52-1.67(m,2H),1.32-1.52(m,4H). 19 F NMR(376MHz,DMSO-d6)δ ppm-105.95--101.82(m,2 F).

[0165] Example 001B: MS measured values ​​(ESI + )[(M+H) + ]:683.2. 1 H NMR(400MHz,DMSO-d6)δ ppm7.87-8.00(m,2H),7.73-7.83(m,2H),6.76-6.83(m,1H),6.69-6.75(m,1H),6 .14-6.32(m,1H),4.37-4.50(m,1H),4.18-4.34(m,3H),4.00-4.18(m,1H),3.74-3 .89(m,3H),3.61-3.72(m,4H),3.02-3.13(m,2H),2.90-3.02(m,4H),2.53-2.65(m ,1H),2.09-2.30(m,1H),1.87-2.04(m,2H),1.59-1.81(m,2H),0.98-1.20(m,4H). 19 F NMR(376MHz,DMSO-d6)δ ppm-108.32--100.42(m,2 F).

[0166] Example 002: 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]azetidine-3-yl]propanamide [ka]

[0167] Step 1: Tert-butyl N-[1-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]azetidine-3-yl]carbamate [ka] Compound Int-2-1 (300 mg, 1.02 mmol), N-(azetidine-3-yl)carbamate tert-butyl ester (263.49 mg, 1.53 mmol), and sodium triacetoxyborohydride (648.53 mg, 3.06 mmol) were added to a mixture of DCM (15 mL) and MeOH (5 mL). The reaction mixture was stirred at room temperature for 1 day. LC-MS showed complete conversion of the starting materials. The mixture was then poured into water (50 mL) and extracted with EA (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain crude product 002a (400 mg, 87.08%) as a yellow solid. MS measured values ​​(ESI + )[(M+H) + ]:450.22.

[0168] Step 3: 1-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]azetidine-3-amine [ka] Compound 002a (300 mg, 0.666 mmol) and TFA (1.5 mL, 20.22 mmol) were dissolved in DCM (15 mL). The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated under vacuum. The resulting residue 002b was azeotropically mixed with toluene and used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:350.8.

[0169] Step 4: Tert-butyl N-[2-[[1-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]azetidine-3-yl]amino]-1-methyl-2-oxoethyl]carbamate [ka]

[0170] Compound 002b (130 mg, 0.280 mmol) and 2-(tert-butoxycarbonylamino)propionic acid (66.23 mg, 0.350 mmol) were dissolved in DCM (15 mL). HATU (159 mg, 0.420 mmol) and DIEA (146.71 μL, 0.840 mmol) were added to the stirred solution. The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was diluted with DCM (15 mL) and water (15 mL). The aqueous layer was extracted with DCM (50 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain crude 002c (120 mg, yield 82.19%) as a yellow solid, which was used directly in the next step.

[0171] Step 5: Tert-butyl N-[2-[[1-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidine-3-yl]amino]-1-methyl-2-oxo-ethyl]carbamate [ka] Compound 002c (120 mg, 0.230 mmol) was dissolved in DMF (10 mL), and N-[(3R)-5-keto-1-(4-piperazinosulfonylphenyl)pyrrolidine-3-yl]carbamate tert-butyl ester (97.7 mg, 0.230 mmol) and DIEA (120 μL, 0.690 mmol) were added. The mixture was prepared using 125 ℃The mixture was stirred overnight. After the starting materials were completely converted, the reaction mixture was diluted with water (50 mL) and extracted with EA (50 mL x 2). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, concentrated to dryness, and crude 002d (150 mg, yield 71.67%) was obtained as a yellow oil, which was used directly in the next step. MS measured value (ESI + )[(M+H) + ]:909.6.

[0172] Step 6: 2-Amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]azetidine-3-yl]propanamide (002) [ka] Compound 002d (100 mg, 0.110 mmol) was dissolved in DCM (5 mL), followed by the addition of TFA (2 mL, 25.96 mmol). The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated under vacuum, and the resulting residue was purified by preparative HPLC to obtain Example 002 (7.3 mg, yield 7.26%) as a pale yellow solid. MS measurement values ​​(ESI) + )[(M+H) + ]:709.3. 1H NMR(400MHz, methanol-d4)δ ppm7.79-7.89(m,2H),7.68-7.78(m,2H),7.39-7.49(m,1H),6.50-6.64(m,1H),4.35-4.46 (m,1H),4.20-4.32(m,1H),3.99-4.11(m,1H),3.86-3.97(m,1H),3.72-3.86(m,2H),3.56- 3.63(m,4H),3.32-3.42(m,1H),3.01-3.11(m,1H),2.88-3.01(m,4H),2.47-2.62(m,1H),2 .00-2.13(m,1H),1.86-1.96(m,1H),1.72-1.82(m,1H),1.59-1.72(m,2H),1.33-1.50(m,8 H),1.13-1.33(m,3H). 19 1F NMR (376MHz, methanol-d4) δ ppm -113.96–-100.71 (m, 2F).

[0173] Example 003: (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Step 1: Benzyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]amino]propyl]carbamate [ka] To a solution of Int-3-1 (313 mg, 1.06 mmol) in DCM (5 mL), benzyl (3-oxopropyl) carbamate (330 mg, 1.59 mmol) and sodium triacetoxyhydroborate (674 mg, 3.18 mmol) were added. The reaction mixture was stirred overnight at 25°C. After completion, the reaction mixture was poured into water and extracted with DCM (50 mL x 3). The combined organic layer was concentrated under vacuum to obtain crude 003a (500 mg, yield 96%) as a yellow oil, which was used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:486.1.

[0174] Step 2: Tert-butyl N-[3-(benzyloxycarbonylamino)propyl]-N-[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]carbamate [ka] Compound 003a (86 mg, 177 μmol) was dissolved in DCM (10 mL). B℃2O (117 mg, 124 μl, 535 μmol) and triethylamine (112 μl, 802 μmol) were added to the stirred solution. The mixture was stirred overnight at room temperature. After completion, the reaction mixture was poured into water and extracted with DCM (50 mL x 3). The combined organic layer was concentrated under vacuum to obtain crude 003b, which was used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:586.1.

[0175] Step 3: Tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[3-(benzyloxycarbonylamino)propylamino]cyclohexyl]difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compounds N-[(3R)-5-keto-1-(4-piperazinosulfonylphenyl)pyrrolidine-3-yl]carbamate tert-butyl ester (100 mg, 0.236 mmol) and 003b (150 mg, 0.256 mmol) were dissolved in DMF (5 mL). DIEA (134 μL, 0.767 mmol) was added to this solution. The reaction mixture was 120 ℃ The mixture was stirred overnight. After completion, the reaction mixture was cooled to room temperature and diluted with EA (100 mL). The aqueous layer was extracted with EA (150 mL x 3). The combined organic layers were washed with water (100 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain crude O03c (200 mg, 80.24%) as a light brown oily substance, which was used directly in the next step.

[0176] Step 4: (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Compound 003c (100 mg, 0.103 mmol) was mixed with TFA (2 mL). The mixture was then 90 ℃ The mixture was heated for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain a crude residue, which was purified by preparative HPLC to obtain Example 003 (2.5 mg, yield 3.43%) as a white solid. MS measured value (ESI + )[(M+H) + ]:640.4. 1H NMR(400MHz, methanol-d4)δ ppm7.89-7.99(m,2H),7.79-7.89(m,2H),7.48-7.62(m,1H),6.60-6.75(m,1H),5.29-5.39(m,1H),4.34- 4.45(m,1H),4.12-4.25(m,1H),3.88-3.99(m,1H),3.62-3.75(m,4H),3.18-3.24(m,1H),2.99-3.18(m,8 H),2.14-2.30(m,3H),1.99-2.14(m,3H),1.76-1.99(m,3H),1.54-1.76(m,2H),1.37-1.52(m,2H). 19 1F NMR (376MHz, methanol-d4) δ ppm -112.27–-105.22 (m, 2F).

[0177] Example 004: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (004A) and cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (004B) [ka] [ka] Step 1: Tert-butyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoromethyl]cyclohexyl]amino]-3-oxopropyl]carbamate [ka] To a solution of Int-3-1 (0.1 g, 339 μmol) in DCM (1.5 mL), 3-((tert-butoxycarbonyl)amino)propanoic acid (0.128 g, 678 μmol) was added, followed by HATU (0.193 g, 509 μmol) and DIEA (177 μL, 1.02 mmol). The reaction mixture was heated overnight at 100 °C. After completion, the reaction mixture was diluted with EA and washed with water (50 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain crude 004a (150 mg, yield 94.94%), which was used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:366.8.

[0178] Step 2: Tert-butyl N-[3-[[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-3-oxo-propyl]carbamate [ka] Compound 004a (150 mg, 322 μmol) and tert-butyl(R)-(5-oxo-1-(4-(piperazine-1-ylsulfonyl)phenyl)pyrrolidine-3-yl)carbamate (100 mg, 236 μmol) were mixed in DMSO (5 mL). DIEA (123 μl, 707 μmol) was added to the stirred suspension. The reaction mixture was stirred overnight at 120°C. After the reaction was complete, it was cooled to room temperature and diluted with EA (50 mL). The organic layer was collected, washed with water (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain crude 004b (150 mg, 176 μmol, yield 74.5%) as a light brown oily substance. MS measurement (ESI + )[(M+HB℃) + ]:854.3.

[0179] Step 3: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (004A) and cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (004B) [ka] [ka] Compound 004b (100 mg, 117 μmol) was dissolved in DCM (5 mL), followed by the addition of TFA (2 mL, 58.5 μmol). The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated under vacuum to obtain a crude residue, which was purified by preparative HPLC and then chiral separated by SFC to obtain the Singer isomers: trans Example 004A (first eluted, 28.5 mg, yield 74.45%) as a white solid and cis Example 004B (second eluted, 28 mg, yield 73.14%) as a white solid. The stereochemistry was determined by 2D NMR.

[0180] Example 004A: MS measured value (ESI + )[(M+H) + ]:654.118. 11H NMR (400 MHz, DMSO-d6) δ ppm 7.97 - 8.08 (m, 1H), 7.88 - 7.96 (m, 2H), 7.77 - 7.86 (m, 2H), 6.76 - 6.82 (m, 1H), 6.66 - 6.74 (m, 1H), 4.21 - 4.36 (m, 1H), 4.00 - 4.16 (m, 1H), 3.77 - 3.90 (m, 1H), 3.58 - 3.69 (m, 4H), 3.03 - 3.14 (m, 1H), 2.89 - 3.03 (m, 6H), 2.34 - 2.44 (m, 2H), 2.08 - 2.24 (m, 1H), 1.75 - 1.87 (m, 2H), 1.58 - 1.68 (m, 2H), 1.02 - 1.34 (m, 6H). 19 19F NMR (376 MHz, DMSO-d6) δ ppm -107.75 - -100.42 (m, 2 F).

[0181] Example 004B: MS measured value (ESI + ) [(M + H) + : 654.118. 1 1H NMR (400 MHz, DMSO-d6) δ ppm 7.99 - 8.07 (m, 1H), 7.87 - 7.98 (m, 2H), 7.75 - 7.84 (m, 2H), 6.72 - 6.89 (m, 1H), 6.60 - 6.75 (m, 1H), 4.20 - 4.32 (m, 1H), 4.03 - 4.16 (m, 1H), 3.74 - 3.91 (m, 2H), 3.60 - 3.74 (m, 4H), 3.02 - 3.13 (m, 1H), 2.85 - 3.02 (m, 4H), 2.55 - 2.71 (m, 3H), 2.08 - 2.28 (m, 1H), 1.61 - 1.80 (m, 2H), 1.35 - 1.56 (m, 5H), 0.96 - 1.35 (m, 3H). 19 19F NMR (376 MHz, DMSO-d6) δ ppm -111.70 - -95.35 (m, 2 F).

[0182] Example 005: cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (005A) and trans-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide (005B) [ka] [ka] Compounds 005A and 005B were prepared in the same manner as the preparations of compounds 004A and 004B, by using (tert-butoxycarbonyl)alanine instead of 3-((tert-butoxycarbonyl)amino)propanoic acid in step 1. Examples 005A (4.3 mg) and 005B (5.7 mg) were obtained as white solids. Their stereochemistry was determined by 2D NMR.

[0183] Example 005A: MS measured value (ESI + )[(M+H) + ]:654.3. 1 1H NMR (400 MHz, methanol-d4) δ ppm7.77-7.91(m,2H),7.65-7.75(m,2H),6.51-6.63(m,2H),4.40-4.59(m,2 H),3.91-4.11(m,2H),3.77-3.90(m,1H),3.64-3.72(m,1H),3.55-3.64(m,4H ),3.46-3.55(m,1H),3.27-3.41(m,1H),2.91-3.03(m,4H),2.74-2.87(m,1H) ,2.04-2.36(m,2H),1.85-1.99(m,3H),1.64-1.79(m,2H),1.30-1.58(m,4H). 191F NMR (376MHz, methanol-d4) δ ppm -109.16 to -99.58 (m, 2F).

[0184] Example 005B: MS measured value (ESI + )[(M+H) + ]:654.3. 1 H NMR(400MHz, methanol-d4)δ ppm7.76-7.89(m,2H),7.64-7.75(m,2H),6.46-6.66(m,2H),4.40-4.61(m,1H),3 .99-4.09(m,1H),3.62-3.70(m,1H),3.54-3.61(m,4H),3.34-3.56(m,3H),2.92-3 .02(m,4H),2.72-2.86(m,1H),2.24-2.38(m,1H),2.06-2.24(m,1H),1.90-2.01(m ,1H),1.76-1.90(m,2H),1.63-1.73(m,2H),1.30-1.55(m,2H),1.04-1.29(m,4H). 19 F NMR (methanol - d4, 376 MHz) δ-106.71 (br d, 2 F, J = 13.6 Hz)

[0185] Example 006: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide [ka] Compound 006 was prepared in the same manner as the preparation of Compound 004A, by using trans-(tert-butoxycarbonylamino)cyclobutanecarboxylic acid instead of 3-((tert-butoxycarbonylamino)cyclobutaneic acid in step 1 without chiral separation. Example 006 (76 mg) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:680.2. 11H NMR (400 MHz, methanol-d4) δ ppm7.80-7.88(m,2H),7.68-7.80(m,2H),6.52-6.64(m,2H),4.23-4.36(m,1 H),4.02-4.15(m,1H),3.78-3.96(m,2H),3.51-3.67(m,4H),3.36-3.51(m,1H ),3.01-3.14(m,2H),2.89-3.01(m,4H),2.50-2.66(m,1H),2.34-2.50(m,2H) ,2.16-2.34(m,2H),1.60-2.05(m,4H),1.31-1.54(m,3H),1.00-1.31(m,4H). 19 1F NMR (376MHz, methanol-d4) δ ppm -113.39–-101.55 (m, 2F).

[0186] Example 007: cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide [ka] Compound 007 was prepared in the same manner as the preparation of Compound 004A, by using cis-(tert-butoxycarbonylamino)cyclobutanecarboxylic acid instead of 3-((tert-butoxycarbonylamino)cyclobutaneic acid in step 1 without chiral separation. Example 007 (47.5 mg) was obtained as a white solid. MS measurement value (ESI + )[(M+H) + ]:680.2. 1H NMR(400MHz, methanol-d4)δ ppm7.89-8.00(m,2H),7.78-7.89(m,2H),6.66-6.73(m,2H),4.36-4.47(m,1H),4 .14-4.29(m,1H),3.89-4.03(m,1H),3.65-3.79(m,5H),3.47-3.65(m,1H),3.15-3 .27(m,1H),3.00-3.15(m,5H),2.85-3.00(m,1H),2.65-2.77(m,1H),2.50-2.65(m ,2H),2.22-2.36(m,2H),1.71-2.22(m,4H),1.43-1.63(m,3H),1.07-1.43(m,4H). 19 1F NMR (376MHz, methanol-d4) δ ppm -115.37 -- 97.60 (m, 2F).

[0187] Example 008: cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-hydroxy-propanamide [ka] Compound 008 was prepared in the same manner as the preparation of Compound 004A, by using Int-4-1 instead of Int-3-1 in Step 1 and 3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9-ylmethoxycarbonylamino)propionic acid instead of 3-((tert-butoxycarbonyl)amino)propanoic acid. Example 008 (3.15 mg) was obtained as a white solid. MS measurement value (ESI + )[(M+H) + ]:670.3. 1H NMR(400MHz, methanol-d4)δ ppm7.79-7.89(m,2H),7.69-7.79(m,2H),6.54-6.63(m,2H),5.22-5.28(m,1H),4 .25-4.36(m,1H),4.01-4.14(m,1H),3.73-3.94(m,4H),3.64-3.73(m,1H),3.52-3 .64(m,4H),3.42-3.52(m,1H),3.04-3.14(m,2H),2.92-3.04(m,4H),2.49-2.64(m ,1H),2.03-2.18(m,2H),1.86-2.03(m,2H),1.69-1.86(m,2H),1.33-1.62(m,2H). 19 1F NMR (376MHz, methanol-d4) δ ppm -109.45–-104.93 (m, 1 F).

[0188] Example 009: cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]urea [ka] Step 1: cis-tert-butyl N-[2-[[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]carbamoylamino]ethyl]carbamate [ka] To a solution of triphosgene (60.32 mg, 0.203 mmol) in acetonitrile (5 mL) cooled in an ice / water bath, solutions of tert--butyl N-(2-aminoethyl) carbamate (96.54 μL, 0.610 mmol) and Et3N (51.42 mg, 70.83 μL, 0.508 mmol) in acetonitrile (5 mL) were added dropwise. Then, a solution of Int-4-1 (60 mg, 0.203 mmol) in acetonitrile (5 mL) was added to the stirred mixture. The reaction mixture was slowly warmed to room temperature and stirred overnight. After completion, the reaction mixture was concentrated under vacuum to obtain crude product 009a (80 mg, yield 81.76%) as a yellow solid, which was used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:481.227.

[0189] Step 2: cis-tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-(tert-butoxycarbonylamino)ethylcarbamoylamino]cyclohexyl]-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compound 009a (83.72 mg, 0.174 mmol) was dissolved in DMF (10 mL), and tert-butyl N-[(3R)-5-oxo-1-(4-piperazine-1-ylsulfonylphenyl)pyrrolidine-3-yl]carbamate (73.83 mg, 0.174 mmol) and DIEA (30 μL, 0.174 mmol) were added at room temperature. The mixture was prepared using 125 ℃ The mixture was stirred overnight. After the starting materials were completely converted, the reaction mixture was diluted with water (50 mL) and extracted with EA (50 mL x 2). The combined organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain crude 009b (100 mg, 66.13%) as a pale yellow solid. MS measured value (ESI + )[(M+H) + ]:869.3.

[0190] Step 3: cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]urea [ka] Compound 009b (100 mg, 0.115 mmol) was dissolved in DCM (10 mL). TFA (1 mL) was added to the stirred solution. The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated under vacuum to obtain a crude residue, which was purified by preparative HPLC to obtain Example 009 (2.5 mg, 2.64%) as a pale yellow solid. MS measured value (ESI + )[(M+H) + ]:669.245. 1 H NMR(400MHz, methanol-d4)δ ppm7.80-7.87(m,2H),7.67-7.78(m,2H),6.48-6.63(m,2H),4.23-4.33(m,1H),4.00- 4.14(m,1H),3.77-3.89(m,1H),3.69-3.77(m,1H),3.52-3.66(m,4H),3.25-3.29(m,1H ),3.02-3.13(m,2H),2.94-3.02(m,4H),2.82-2.94(m,2H),2.45-2.68(m,1H),2.07-2. 20(m,1H),1.86-2.07(m,3H),1.63-1.80(m,2H),1.40-1.55(m,5H),1.27-1.40(m,2H). 19 F NMR (376 MHz, methanol-d4) δ ppm -115.65–-100.71 (m, 2 F).

[0191] Example 010: Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5,5-dimethyl-5-azoniaspiro[2,4]heptan-6-carboxamide [ka] Step 1: (6S)-5,5-dimethyl-5-azoniaspiro[2,4]heptane-6-carboxylic acid [ka] (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid (290 mg, 2.05 mmol) was dissolved in DMF (10 mL), and iodomethane (1.28 mL, 20.54 mmol) and DIEA (3.6 mL, 20.54 mmol) were added. The reaction mixture was stirred overnight at room temperature. After completion, the reaction mixture was concentrated to dryness to obtain crude product 010a (300 mg, 85.79%) as a pale yellow oil, which was used directly in the next step.

[0192] Step 2: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[[4-[[(6S)-5,5-dimethyl-5-azoniaspiro[2,4]heptan-6-carbonyl]amino]cyclohexyl]difluoro-methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] Compound Int-5-1 (150 mg, 0.220 mmol) was dissolved in DMF (10 mL), and then compounds 010a (100 mg, 0.587 mmol), HATU (208.7 mg, 0.549 mmol), and DIEA (85.12 mg, 115.03 μL, 0.659 mmol) were added at room temperature. The mixture was stirred overnight at room temperature. After completion, the reaction mixture was diluted with saturated NH4Cl aqueous solution (50 mL) and extracted with EA (50 mL × 2). The combined organic layer was washed with water (50 mL × 3) and brine (50 mL), dried over anhydrous Na2SO4, concentrated to dryness, and crude 010b (180 mg, yield 98.14%) was obtained as a pale yellow oil, which was used directly in the next step. MS measured values ​​(ESI + )[(M+H) + ]:834.6.

[0193] Step 3: Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5,5-dimethyl-5-azoniaspiro[2,4]heptan-6-carboxamide [ka] Compound 010b (150 mg, 0.180 mmol) was dissolved in DCM (5 mL), followed by the addition of TFA (1.5 mL, 19.47 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated to dryness to obtain the crude substance, which was purified by preparative HPLC to obtain Example 010 (26 mg, yield 18.71%) as a white solid. MS measurement values ​​(ESI) + )[(M+H) + ]:734.4. 1 H NMR(500MHz,DMSO-d6)δ ppm8.58-8.71(m,1H),8.27-8.41(m,2H),7.87-7.98(m,2H),7.73-7.84(m,2H),6.76-6.85(m,1H),6.66- 6.76(m,1H),4.35-4.43(m,1H),4.21-4.33(m,1H),4.04-4.13(m,1H),3.78-3.90(m,2H),3.63-3.70(m,4H) ),3.20-3.29(m,3H),3.10-3.20(m,4H),3.02-3.10(m,1H),2.92-3.02(m,4H),2.55-2.64(m,2H),2.25-2. 45(m,2H),2.07-2.25(m,1H),1.76-1.98(m,2H),1.57-1.76(m,2H),1.04-1.34(m,4H),0.58-0.89(m,4H). 19 F NMR(471MHz,DMSO-d6)δ ppm-126.70--92.41(m,2 F).

[0194] Example 011: trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5-methyl-5-azoniaspiro[2,4]heptan-6-carboxamide(011 A) and trans-(5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptan-6-carboxamide (011B) [ka] [ka]

[0195] Step 1: (6S)-5-(2-amino-2-oxo-ethyl)-5-azaspiro[2,4]heptane-6-carboxylic acid [ka] To a solution of (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid (100 mg, 0.708 mmol) in acetonitrile (5 mL), 3-bromopropionamide (161.5 mg, 1.06 mmol) and K2CO3 (293.71 mg, 2.13 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was diluted with water (50 mL). The aqueous layer was extracted with EA (50 mL x 2). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, concentrated to dryness, and crude O11a (120 mg, yield 85.46%) was obtained as a pale yellow solid, which was used directly in the next step.

[0196] Step 2: Trans-tert-butyl N-[(3R)-1-[4-[4-[[4-[[4-[[(6S)-5-(2-amino-2-oxo-ethyl)-5-azaspiro[2,4]heptan-6-carbonyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of compound Int-5 (150 mg, 0.220 mmol) in DMF (10 mL), 011a (116.44 mg, 0.587 mmol), HATU (208.7 mg, 0.549 mmol), and DIEA (85.12 mg, 115.03 μL, 0.659 mmol) were added. The reaction mixture was stirred overnight at room temperature. After completion, the reaction mixture was diluted with saturated NH4Cl solution (100 mL). The aqueous layer was extracted with EA (100 mL x 2). The organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4, and concentrated to dryness. The crude product was purified by silica gel chromatography (0-10% MeOH / DCM) to obtain 011b (150 mg, yield 79.13%) as a pale yellow solid. MS measured value (ESI + )[(M+H) + ]:863.6.

[0197] Step 3: Trans-tert-butyl N-[(3R)-1-[4-[4-[[4-[[4-[[(6S)-5-(2-amino-2-oxo-ethyl)-5-methyl-5-azoniaspiro[2,4]heptan-6-carbonyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of compound 011b (150 mg, 0.174 mmol) in DMF (5 mL), iodomethane (21.73 μL, 0.347 mmol) and DIEA (151.71 μL, 0.869 mmol) were added. The reaction mixture was stirred overnight at room temperature. After completion, the reaction mixture was concentrated to dryness, yielding crude product 011c (150 mg, yield 98.29%) as a dark brown oily substance, which was used directly in the next step. MS measurement values ​​(ESI + )[(M+H) + ]:877.664.

[0198] Step 4: Trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5-methyl-5-azoniaspiro[2,4]heptan-6-carboxamide(011A ) and trans-(5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptan-6-carboxamide(011B) [ka] [ka] To a solution of O11c (150 mg, 0.171 mmol) in DCM (5 mL), TFA (1.5 mL, 19.47 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated to dryness. The crude product was purified by preparative HPLC to obtain O11A (first to elute, 8.1 mg, 5.49%) and O11B (second to elute, 10.1 mg, 6.84%) as white solids.

[0199] Example 011A: MS measured value (ESI + )[(M+H)+ :777.4. 1 1H NMR (500 MHz, DMSO-d6) δ ppm 7.92 (d, J = 8.70 Hz, 2H), 7.80 (d, J = 8.70 Hz, 2H), 6.79 (s, 1H), 6.73 (s, 1H), 4.57 - 4.75 (m, 1H), 4.18 - 4.37 (m, 3H), 4.07 (br d, J = 11.14 Hz, 2H), 3.75 - 3.89 (m, 2H), 3.66 (br s, 4H), 3.17 (s, 2H), 3.08 (br dd, J = 17.85, 8.24 Hz, 2H), 2.98 (br s, 4H), 2.54 - 2.62 (m, 2H), 2.40 - 2.49 (m, 1H), 2.15 (br dd, J = 13.73, 4.43 Hz, 2H), 1.87 (br d, J = 10.68 Hz, 1H), 1.74 - 1.83 (m, 1H), 1.60 - 1.73 (m, 2H), 1.10 - 1.26 (m, 4H), 0.72 - 0.91 (m, 4H).<0000​​​​​​​​​​​​F NMR(471MHz,DMSO-d6)δ ppm-114.06--91.20(m,2 F).

[0201] Example 12: (R)-4-amino-1-(4-((4-(6-chloro-4-(difluoro((1r,4r)-4-(3-hydroxyazetidine-1-yl)cyclohexyl)methyl)pyridine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one [ka] Step 1: Tert-butyl((R)-1-(4-((4-(6-chloro-4-(difluoro((1r,4r)-4-(3-hydroxyazetidine-1-yl)cyclohexyl)methyl)pyridine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] To a solution of compound Int-2 (100.0 mg, 0.1 mmol) in NMP (3 mL), 1,3-dibromo-2-propanol (32.0 mg, 0.1 mmol) and DIEA (54.0 mg, 0.4 mmol) were added at 25°C. The reaction mixture was stirred at 50°C for 2 hours. After cooling to room temperature, the resulting mixture was poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. This was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 12a (74.0 mg, 0.1 mmol, yield 70.0%) was obtained as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:740.1

[0202] Step 2: (R)-4-amino-1-(4-((4-(6-chloro-4-(difluoro((1r,4r)-4-(3-hydroxyazetidine-1-yl)cyclohexyl)methyl)pyridine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one [ka] To a solution of compound 12a (40.0 mg, 0.1 mmol) in DCM (1 mL), TFA (1.0 mL, 12.98 mmol) was added at 25°C, and the mixture was stirred at 25°C for 2 hours. After concentrating the mixture under reduced pressure, the residue was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 19%~49%, 10 min), and lyophilized to obtain Example 12 (21.2 mg, 0.1 mmol, yield 54.5%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:640.3.

[0203] Example 41: (4R)-4-amino-1-[4-[4-[4-[[4-(azetidine-1-yl)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Example 41 was prepared in the same manner as the preparation of compound Example 12, except that 1,3-dibromopropane was used instead of 1,3-dibromo-2-propanol in step 1. Example 41 (43.3 mg, 0.1 mmol, yield 49.0%) was obtained as a white solid in the form of a TFA salt. MS measured value (ESI+)[({ 35 Cl}M+H) + ]:623.1.

[0204] Example 13: cis-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Step 1: Tert-butyl N-[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] To a solution of compounds Int-3 (214.8 mg, 0.5 mmol) and Int-1 (200.0 mg, 0.5 mmol) in DMSO (2 mL), DIEA (0.5 mL, 0.51 mmol) was added at 25 °C, and the mixture was heated to 110 °C with stirring for 12 hours. After cooling to room temperature, Depositphotos (100 mL) and water (30 mL) were added to the mixture. The resulting mixture was transferred to a separatory funnel. The aqueous layer was extracted with Depositphotos (50 mL x 2). The combined organic layers were washed with saturated brine (80 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was separated into TLC (PE / Depositphotos = 1 / 1; TLC: PE / Depositphotos = 1 / 1, R f The compound was purified by (=0.4) to obtain compound 13a (280.0 mg, 0.4 mmol, yield 70.6%) as a yellow solid. MS measured value (ESI + )[{ 35 Cl}(M+H) + ]:783.4.

[0205] Step 2: cis-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] To a solution of compound 13a (250.0 mg, 0.3 mmol) in DCM (3 mL), TFA (3 mL) was added at 25°C and stirred at this temperature for 2 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was redissolved in water (10 mL), and 5% (w%) NH3·H2O aqueous solution (10 mL) was added to the mixture. A solid was formed, and the mixture was stirred at 20°C for 16 hours. The resulting precipitate was filtered. The filtered cake was collected, dissolved in water (20 mL), and the suspension was freeze-dried to obtain 13 (153.4 mg, 0.3 mmol, yield 81.5%) as a white solid. MS measured value (ESI + )[{ 35 Cl}(M+H) + ]:583.0.

[0206] Example 14: trans-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Step 1: Tert-butyl-N-[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] To a solution of compound Int-2 (200.0 mg, 0.3 mmol) in DCM (5 mL), TFA (2.0 mL, 26.0 mmol) was added at 20°C, and the mixture was stirred at this temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 17%~47%, 10 min), lyophilized, and the product (100.0 mg) was obtained in the form of the HCl salt. The product was dissolved in water (10 mL) and 5% (w%) aqueous NH3·H2O (10 mL). The solid was stirred at 20°C for 16 hours, precipitated, and filtered. The filtered cake was collected and redissolved in water (20 mL). The suspension was freeze-dried to obtain 14 (55.3 mg, 0.1 mmol, yield 35.4%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:583.1.

[0207] Example 15: 2-Amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]acetamide [ka] Step 1: Tert-butyl N-[2-[[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-4-pyridyl]-difluoro-methyl]cyclohexyl]methylamino]-2-oxo-ethyl]carbamate [ka] To a solution of compound Int-4 (120.0 mg, 0.2 mmol) and (tert-butoxycarbonyl)glycine (20.7 mg, 0.1 mmol) in DMF (1 mL), HATU (129.2 mg, 0.3 mmol) and DIEA (70.0 mg, 0.5 mmol) were added at 25°C. The reaction mixture was stirred at 25°C for 2 hours, and the resulting mixture was diluted with H2O (30 mL). The mixture was transferred to a separatory funnel, and the aqueous layer was extracted with SiO (50 mL x 3). The combined organic layers were washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain 15a (40.0 mg, 0.1 mmol, yield 59.4%) as a white solid. MS measured values ​​(ESI) + )[{ 35 Cl}(M+H) + ]:854.3.

[0208] Step 3: 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]acetamide [ka] To a solution of compound 15a (40.0 mg, 0.1 mmol) in DCM (1 mL), TFA (1.0 mL, 13.0 mmol) was added at 25°C, and the mixture was stirred at 25°C for 1 hour. The mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 18%~48%, 10 min), and lyophilized to obtain Example 15 (18.0 mg, 0.1 mmol, yield 57.5%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:654.3.

[0209] Example 16: 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N-(carboxymethyl)-4-oxobutan-1-aminium [ka] Step 1: Benzyl 4-(bis(3-((tert-butoxycarbonyl)amino)propyl)amino)butanoate [ka] To a solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (2.5 g, 7.7 mmol) in ACN (15 mL), benzyl 4-bromobutanoate (2.0 g, 7.7 mmol) and NaHCO3 (1.0 g, 11.5 mmol) were added at 25°C, and the mixture was stirred at 25°C for 16 hours. After filtering the mixture, the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 16a (1.8 g, 3.6 mmol, yield 46.2%) was obtained as a colorless oil. MS measurement value (ESI + )[(M+H) + ]:508.1.

[0210] Step 2: 4-(benzyloxy)-N-(2-(tert-butoxy)-2-oxoethyl)-N,N-bis(3-((tert-butoxycarbonyl)amino)propyl)-4-oxobutane-1-aminium bromide [ka] To a solution of compound 16a (1.0 g, 2.0 mmol) in MeCN (3 mL), tert-butylbromoacetate (461.1 mg, 2.4 mmol) and NaHCO3 (413.7 mg, 4.9 mmol) were added at 25°C with stirring for 16 hours. After filtering the mixture, the filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 16b (640.0 mg, 1.0 mmol, yield 52.2%) was obtained as a colorless oil. MS measurement value (ESI + )[(M+H) + ]:622.3.

[0211] Step 3: N-(2-(tert-butoxy)-2-oxoethyl)-3-((tert-butoxycarbonyl)amino)-N-(3-((tert-butoxycarbonyl)amino)propyl)-N-(3-carboxypropyl)propane-1-aminium bromide [ka] A solution of compound 16b (650.0 mg, 1.0 mmol) in isopropanol (10 mL) was mixed with wet Pd / C (106.4 mg, 5% wt, containing 50% water) under an N2 atmosphere. The suspension was degassed under vacuum, purged three times with H2, and stirred at 25°C for 3 hours under an H2 atmosphere. After dilution with MeOH (100 mL), the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 16c (550.0 mg, 1.0 mmol, yield 98.9%) as a white solid. The crude product was used directly in the next step without further purification. MS measured values ​​(ESI + )[(M+H) + ]:532.4.

[0212] Step 4: 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N-(carboxymethyl)-4-oxobutan-1-aminium (Example 16) [ka] Example 16 was prepared in the same manner as in Example 15, except that Int-2 and 16c were used instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively, in step 1. Example 16 (102.0 mg, 0.1 mmol, yield 76.3%) was obtained as a pale yellow solid in the form of a hydrochloride salt. MS measured values ​​(ESI) + )[({ 35 Cl}M+H) + ]:840.1.

[0213] Example 17: N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-(2-aminoethoxy)acetamide [ka] Example 17 was prepared in the same manner as the preparation of compound Example 15, by using Int-2 and 2-[2-[(2-methylpropan-2-yl)oxycarbonylamino]ethoxy]acetic acid instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively, in step 1, and by using T3P instead of HATU. Example 17 (12.2 mg, 0.1 mmol, yield 35.7%) was obtained as a white solid in the form of a hydrochloride salt. MS measurement values ​​(ESI + )[({ 35 Cl}M+H) + ]:684.2.

[0214] Example 18: 3-Amino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-hydroxypropanamide [ka] Example 18 was prepared in the same manner as the preparation of compound Example 15, by using Int-2 and 3-(((benzyloxy)carbonyl)amino)-2-hydroxypropanoic acid instead of Int-4 and (tert-butoxycarbonyl)glycine in step 1, respectively. Example 18 (6.0 mg, 0.01 mmol, yield 22.4%) was obtained as a white solid in the form of a hydrochloride salt. MS measured value (ESI+)[({ 35 Cl}M+H) + ]:670.2.

[0215] Example 19: 2,3-diamino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)propanamide [ka] Example 19 was prepared in the same manner as the preparation of compound Example 15, except that in step 1, Int-2 and 2-(((benzyloxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid were used instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively. Example 19 (14.0 mg, 0.02 mmol, yield 41.8%) was obtained as a white solid in the form of a hydrochloride salt. MS measurement values ​​(ESI + )[({ 35 Cl}M+H) + ]:669.2.

[0216] Example 26: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide [ka] Step 1: Benzyl 4-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)butanoate [ka] To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (3.0 g, 9.9 mmol) in ACN (10 mL), benzyl 4-bromobutanoate (2.8 g, 10.9 mmol) and potassium carbonate (6.8 g, 49.4 mmol) were added at 20 °C, and the mixture was heated to 45 °C with stirring for 12 hours. After cooling to room temperature, the mixture was diluted with water (100 mL), extracted with ELISA (100 mL x 3), washed with saturated brine (100 mL), dried over anhydrous sodium 2 SO4, and filtered. The filtrate was concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (PE: Depositphotos = 1:0 to 0:1; TLC: PE / Depositphotos = 3 / 1, Rf = 0.5) to obtain 26a (1.6 g, 3.3 mmol, yield 99.0%) as a colorless oil.

[0217] Step 2: 4-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]butanoic acid [ka] To a solution of 26a (800.0 mg, 1.7 mmol) in methanol (8 mL), wet Pd / C (170.0 mg, 5% wt, containing 50% water) was added under N2 at 25°C. The mixture was degassed under vacuum, purged three times with H2, and stirred at 25°C for 3 hours under an H2 atmosphere (15 psi). After diluting the mixture with MeOH (100 mL), it was filtered, and the filtrate was concentrated under vacuum to obtain crude product 26b (600.0 mg, 1.5 mmol, yield 92.4%) as a colorless oil. This crude product was used directly in the next step without further purification. MS measured values ​​(ESI + )([M+H] + ):390.4.

[0218] Step 3: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide (Example 26) [ka] Example 26 was prepared in the same manner as the preparation of compound Example 15, by using Int-2 and 26b instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively, in step 1. Example 26 (30.4 mg, 0.03 mmol, yield 51.0%) was obtained as a yellow solid in the form of the TFA salt. MS measured values ​​(ESI + )([{ 35 Cl}M+H] + ):754.1.

[0219] Example 32: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide [ka] Step 1: Benzyl 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoate [ka] A solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (500.0 mg, 1.5 mmol) and benzyl 4-bromobutanoate (387.9 mg, 1.5 mmol) in ACN (5 mL) was mixed with K2CO3 (615.5 mg, 4.5 mmol) at 20 °C and heated to 40 °C with stirring for 12 hours. The mixture was poured into water (30 mL) and extracted with  (50 mL x 3). The combined organic layer was washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain compound 32a (400.0 mg, 0.8 mmol, yield 52.2%) as a colorless oil. MS measured value (ESI + )[(M+H) + ]:508.4.

[0220] Step 2: 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoic acid [ka] A solution of compound 32a (300.0 mg, 0.6 mmol) in methanol (5 mL) was mixed with wet Pd / C (100.0 mg, 5% wt, containing 50% water) at 20°C. The mixture was degassed under vacuum, purged three times with H2, and stirred at 20°C under an H2 atmosphere (15 psi) for 3 hours. After dilution with MeOH (100 mL), the mixture was filtered, and the filtrate was concentrated under vacuum to obtain crude product 32b (200.0 mg, 0.5 mmol, yield 81.1%) as a colorless oil. The crude product was used directly in the next step without further purification. MS measured values ​​(ESI + )[(M+H)+ ]:418.0.

[0221] Step 3: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide [ka] Example 32 was prepared in the same manner as the preparation of compound Example 15, by using Int-2 and 32b instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively, in step 1. Example 32 (14.7 mg, 0.02 mmol, yield 38.7%) was obtained as a white solid in the form of hydrochloride. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:782.3.

[0222] Example 31: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N-hydroxy-butanamide [ka] Example 31 was prepared in the same manner as the preparation of compound Example 15, by using Int-6 and 32b instead of Int-4 and (tert-butoxycarbonyl)glycine, respectively, in step 1. Example 31 (4.1 mg, 0.01 mmol, yield 26.6%) was obtained as a white solid in the form of hydrochloride. MS measured value (ESI)[({ 35 Cl}M+H) + ]:798.3.

[0223] Example 21: (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-4-oxo-butyl]-bis(3-aminopropyl)ammonium [ka] Step 1: Benzyl 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoate [ka] To a solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (2.5 g, 7.7 mmol) in ACN (15 mL), NaHCO3 (966.5 mg, 11.5 mmol) and benzyl 4-bromobutanoate (2.0 g, 7.7 mmol) were added at 25°C, and the mixture was stirred at 25°C for 16 hours. The mixture was filtered, and the filtrate was concentrated to obtain the residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 21a (3.6 g, 7.1 mmol, yield 92.5%) was obtained as a colorless oil. MS measured value (ESI + )[(M+H) + ]:508.3.

[0224] Step 2: 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoic acid [ka] Compound 21a (400.0 mg, 0.8 mmol) was added to a suspension of wet Pd / C (20.0 mg, 5% wt, containing 50% water) in methanol (5 mL) under N2 at 25°C. The mixture was degassed, purged three times with H2, and stirred at 25°C for 16 hours using an H2 balloon. The mixture was diluted with MeOH (100 mL), filtered, and the filtrate was concentrated to obtain crude product 21b (330.0 mg, 0.8 mmol) as a colorless, rubbery substance. MS measurement values ​​(ESI) + )[(M+H) + ]:418.2.

[0225] Step 3: tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propyl-[4-[[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-4-oxo-butyl]amino]propyl]carbamate [ka] To a solution of 21b (300.0 mg, 0.7 mmol) in DMF (2 mL), DIEA (0.4 mL, 2.2 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (409.8 mg, 1.1 mmol) were added under N2 conditions at 0°C. After stirring at 0°C for 0.5 hours, Int-2 (490.9 mg, 0.7 mmol) was added, and the mixture was stirred at 25°C for 1 hour. This mixture was concentrated in a vacuum to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 21c (500.0 mg, 0.5 mmol, yield 61.1%) was obtained as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:1082.6.

[0226] Step 4: (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]amino]-4-oxo-butyl]-bis[3-(tert-butoxycarbonylamino)propyl]ammonium; iodide [ka] To a solution of 21c (150.0 mg, 0.1 mmol) in ACN (2 mL), iodoacetamide (153.7 mg, 0.8 mmol) and DIEA (0.1 mL, 0.8 mmol) were added at 20°C, and the mixture was heated to 40°C with stirring for 16 hours. The mixture was poured into water (30 mL) and extracted with  (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 21d (80.0 mg, 0.1 mmol, yield 50.6%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:1139.6.

[0227] Step 5: 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]azetidine-3-yl]propanamide [ka] To a solution of 21d (80.0 mg, 0.1 mmol) in DCM (2 mL), trifluoroacetic acid (1 mL) was added at 25°C and the mixture was stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15%~45%, 10 min), lyophilized, and Example 21 (46.3 mg, 0.1 mmol, yield 82.4%) was obtained as a white solid in the form of hydrochloride. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:839.3.

[0228] Example 20: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(diaminomethylene)urea [ka] Step 1: tert-butyl((R)-1-(4-((4-(((1r,4r)-4-(1H-imidazole-1-carboxamide)cyclohexyl)difluoromethyl)-6-chloropyridine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] A mixture of compound Int-2 (100.0 mg, 0.2 mmol) and N,N'-carbonyldiimidazole (120.0 mg, 0.8 mmol) in anhydrous DCM (3 mL) was mixed with DIEA (100.0 mg, 0.8 mmol) at 20°C and stirred for 12 hours. The mixture was poured into water (30 mL) and extracted with  (30 mL x 3). The combined organic layer was washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain crude 20a (110.0 mg, 0.1 mmol, yield 96.7%) as a white solid, which was used directly in the next step without further purification. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:777.3.

[0229] Step 2: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[[(E)-[amino-(tert-butoxycarbonylamino)methylene]carbamoyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] A mixture of compound 20a (80.0 mg, 0.1 mmol) and potassium carbonate (240.0 mg, 1.7 mmol) in anhydrous DMF (1 mL) was mixed with tert-butyl N-carbamimidylcarbamate (80.0 mg, 0.5 mmol) at 20°C and stirred for 12 hours. The mixture was poured into water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with saturated brine (50 mL x 3), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 20b (15.0 mg, 0.02 mmol, yield 16.8%) was obtained as a yellow solid. MS measured value (ESI + )[(M+H)+ ]:868.3.

[0230] Step 3: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(diaminomethylene)urea [ka] A mixture of compound 20b (15.0 mg, 0.02 mmol) in anhydrous DCM (1 mL) was mixed with trifluoroacetic acid (0.5 mL, 6.5 mmol) at 20°C and stirred for 1 hour. The mixture was concentrated under vacuum to obtain the crude product, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 17%~47%, 10 min), lyophilized, and 20 (1.5 mg, 0.01 mmol, yield 12.1%) was obtained as a yellow solid in the form of the hydrochloride salt. MS measured value (ESI + )[(M+H) + ]:668.2.

[0231] Example 22: 3-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoylamino]propyl-bis(3-aminopropyl)-(carboxymethyl)ammonium [ka] Step 1: tert-butyl N-[3-[3-(benzyloxycarbonylamino)propyl-[3-(tert-butoxycarbonylamino)propyl]amino]propyl]carbamate [ka] A solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (2.0 g, 6.0 mmol) and benzyl N-(3-bromopropyl)carbamate (1.8 g, 6.6 mmol) in ACN (30 mL) was mixed with NaHCO3 (1.5 g, 18.1 mmol) at 25°C and stirred at 25°C for 12 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. This residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 330 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 25 min), lyophilized, and 22a (1.0 g, 1.9 mmol, yield 31.7%) was obtained as a yellow oily substance. MS measured value (ESI + )[(M+H) + ]:523.8.

[0232] Step 2: 3-(benzyloxycarbonylamino)propyl-bis[3-(tert-butoxycarbonylamino)propyl]-(2-tert-butoxy-2-oxo-ethyl)ammonium; bromide [ka] To a solution of 22a (1.0 g, 1.9 mmol) in DMSO (5 mL), NaHCO3 (482.1 mg, 5.7 mmol) and tert-butylbromoacetate (1.5 g, 7.7 mmol) were added at 20°C, and the mixture was stirred at 50°C for 15 hours. The mixture was diluted with siRNA (100 mL) and filtered. The filtrate was concentrated to obtain the residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 22b (500.0 mg, 0.7 mmol, yield 36.4%) was obtained as a yellow oily substance. MS measured value (ESI + )[(M+H) + ]:637.2.

[0233] Step 3: 3-aminopropyl-bis[3-(tert-butoxycarbonylamino)propyl]-(2-tert-butoxy-2-oxo-ethyl)ammonium; bromide [ka] To a suspension of wet Pd(OH)2 / C (0.5 g, 5% wt, containing 50% water) in THF (20 mL), 22b (1.0 g, 1.6 mmol) was added under an N2 atmosphere at 25 °C. The suspension was degassed under vacuum and purged three times with H2. The mixture was then stirred at 50 °C for 2 hours under H2 (15 psi). The mixture was diluted with MeOH (100 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product 22c (500.0 mg, 1.0 mmol, yield 63.3%) as a yellow oil. 22c was used directly in the next step without further purification. MS measured values ​​(ESI + )[(M+H) + ]:503.5. [ka]

[0234] Example 22 was prepared in the same manner as the preparation of compound Example 20, by using 22c instead of tert-butyl N-carbamimidylcarbamate and TEA instead of K2CO3 in step 2. Example 22 (14.0 mg, 0.02 mmol, yield 22.5%) was obtained as a yellow solid in the form of hydrochloride. MS measurement (ESI + )[({ 35 Cl}M+H) + ]:855.5.

[0235] Example 23: 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea [ka] Example 23 was prepared in the same manner as the preparation of compound Example 20, except that in step 2, tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate was used instead of tert-butyl N-carbamimidoylcarbamate and DIEA was used instead of K2CO3. In step 2, Example 23 (16.0 mg, 0.02 mmol, yield 40.0%) was obtained in the form of hydrochloric acid as a white powder. MS measurement (ESI + )[({ 35 Cl}M+H) + ]:740.4.

[0236] Example 34: (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-hydroxy-pyrrolidine-1-carboxamide [ka] Example 34 was prepared in the same manner as the preparation of compound Example 20, except that in step 2, tert-butyl N-[(3R,4S)-4-hydroxypyrrolidine-3-yl]carbamate was used instead of tert-butyl N-carbamimidoylcarbamate and DIEA was used instead of K2CO3. Example 34 (41.6 mg, 0.1 mmol, yield 53.3%) was obtained as a white solid in the form of hydrochloride. MS measurement (ESI + )[({ 35 Cl}M+H) + ]:711.3.

[0237] Example 35: 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2-aminoethyl)amino)ethyl)urea [ka] Step 1: tert-butyl N-[2-[2-(benzyloxycarbonylamino)ethyl-[2-(tert-butoxycarbonylamino)ethyl]amino]ethyl]carbamate [ka] To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (1.0 g, 3.3 mmol) in ACN (10 mL), benzyl N-(2-bromoethyl)carbamate (1.0 g, 4.0 mmol) and NaHCO3 (830.6 mg, 9.9 mmol) were added at 20°C. The mixture was stirred at 20°C for 15 hours, then diluted with ELISA (100 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (PE / ELISA = 10 / 1 to 1 / 1; TLC: PE / ELISA = 3 / 1, Rf = 0.4) to obtain 35a (1.0 g, 2.1 mmol, yield 63.1%) as a colorless oil. MS measured value (ESI + )[(M+H) + ]:481.1.

[0238] Step 2: tert-butyl N-[2-[2-aminoethyl-[2-(tert-butoxycarbonylamino)ethyl]amino]ethyl]carbamate [ka] To a solution of 35a (300.0 mg, 0.6 mmol) in THF (2 mL) and isopropanol (2 mL), dried Pd / C (100.0 mg, 10% by weight) was added at 20°C under an N2 atmosphere. The suspension was degassed under reduced pressure and purged three times with H2. The mixture was then stirred at 20°C for 1 hour under H2 (45 psi). The mixture was diluted with MeOH (200 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain crude 35b (200.0 mg, 0.6 mmol, yield 92.5%) as a yellow oily substance, which was used directly in the next step without further purification. MS measured value (ESI + )[(M+H)+ ]:347.1.

[0239] Step 3: 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2-aminoethyl)amino)ethyl)urea [ka] Example 35 was prepared in the same manner as the preparation of compound Example 20, by using 35b instead of tert-butyl N-carbamimidylcarbamate and DIEA instead of 2CO3 in step 2. Example 35 (34.0 mg, 0.1 mmol, yield 50.0%) was obtained as a white solid in the form of its hydrochloride salt. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:755.2.

[0240] Example 29: (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]morpholine-4-carboxamide [ka] Step 1: Tert-butyl N-(oxiran-2-ylmethyl)carbamate [ka] To a solution of tert-butyl N-allyl carbamate (20.0 g, 127.2 mmol) in DCM (100 mL), m-CPBA (32.8 g, 152.7 mmol) was added at 20°C and the mixture was stirred at 20°C for 1 hour. The reaction product was quenched with aqueous Na2SO3 (100 mL) and the mixture was stirred at 20°C for 1 hour. After extraction with Depositphotos (200 mL x 3), the combined organic layers were washed with saturated aqueous NaHCO3 (50 mL x 3) and saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain crude product 29a (15.0 g, 86.6 mmol, yield 68.1%) as a yellow oil. The crude product was used directly in the next step without further purification.

[0241] Step 2: Tert-butyl N-[3-(benzylamino)-2-hydroxypropyl]carbamate [ka] A solution of compound 29a (18.0 g, 103.9 mmol) and benzylamine (11.4 mL, 103.9 mmol) in isopropanol (100 mL) was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 330 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-70%, 20 min), lyophilized, and compound 29b (12.0 g, 42.8 mmol, yield 41.2%) was obtained as a white solid. MS measured values ​​(ESI) + )[(M+H) + ]:281.1.

[0242] Step 3: Tert-butyl N-[3-[benzyl-(2-chloro-2-cyanoethyl)amino]-2-hydroxypropyl]carbamate [ka] To a solution of compound 29b (15.0 g, 53.5 mmol) in THF (200 mL), 2-chloroacrylonitrile (4.9 g, 56.2 mmol) was added at 0°C, and the mixture was heated to 20°C with stirring for 16 hours. The mixture was poured into water (300 mL) and extracted with siRNA (300 mL x 3). The combined organic layer was washed with saturated brine (300 mL x 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain crude 29c (12.0 g, 32.6 mmol, yield 61.0%) as a pale yellow oil, which was used directly in the next step without further purification. MS measured value (ESI + )[(M+H) + ]:368.1.

[0243] Step 4: Tert-butyl N-[[rac-(2S,6R)-4-benzyl-6-cyanomorpholin-2-yl]methyl]carbamate [ka] [ka] A solution of compound 29c (15.0 g, 40.8 mmol) in THF (100 mL) was added at 0°C to a solution of potassium tert-butoxide in THF (1.0 M, 41.0 mL, 41.0 mmol), and the mixture was heated to 20°C with stirring for 2 hours. The mixture was poured into water (100 mL) and extracted with ELISA (100 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 330 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 30 min), lyophilized, and obtained 29d-1 (5.2 g, 15.7 mmol, yield 27.5%) and 29d-2 (4.1 g, 12.4 mmol, yield 21.7%) as pale yellow oily substances. MS measured values ​​(ESI + )[(M+H) + ]:332.1.

[0244] Step 5: Tert-butyl N-[[6-(aminomethyl)-4-benzylmorpholin-2-yl]methyl]carbamate [ka] A solution of compound 29d-1 (200.0 mg, 0.6 mmol) in THF (3 mL) was mixed with a solution of BH3·Me2S (10.0 M, 0.5 mL, 5.0 mmol) at 0°C and stirred at 20°C for 2 hours. The mixture was carefully quenched with MeOH (50 mL) at 0°C and concentrated under reduced pressure to obtain crude 29e (150.0 mg, 0.5 mmol, yield 74.1%) as a colorless oil, which was used directly in the next step without further purification. MS measured values ​​(ESI + )[(M+H) + ]:336.1.

[0245] Step 6: Tert-butyl N-[[4-benzyl-6-[(tert-butoxycarbonylamino)methyl]morpholine-2-yl]methyl]carbamate [ka] Compound 29e (150.0 mg, 0.5 mmol) was added at 20°C to a solution of DIEA (0.3 mL) and di-tert-butyl dicarbonate (100.0 mg, 0.5 mmol) in THF (1 mL), and the mixture was stirred at this temperature for 12 hours. The reaction mixture was poured into  (50 mL) and water (10 mL). The organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was filtered through a silica gel column (PE / Â=50 / 1~2 / 1; TLC:PE / Â=3 / 1, R f The compound was purified by (=0.4) to obtain compound 29f (150.0 mg, 0.3 mmol, yield 77.0%) as a colorless oil. MS measured value (ESI + )[(M+H) + ]:436.1.

[0246] Step 7: Tert-butyl(((2R,6R)-morpholine-2,6-diyl)bis(methylene))dicarbamate [ka] A solution of compound 29f (150.0 mg, 0.2 mmol) in isopropanol (5 mL) was mixed with wet Pd / C (50.0 mg, 5% wt, containing 50% water) under an N2 atmosphere at 20°C. The suspension was degassed under vacuum, purged three times with H2, and stirred at 20°C under H2 (15 psi) for 12 hours. The mixture was diluted with methanol (200 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain 29 g of crude compound (50.0 mg, 0.2 mmol, yield 36.3%) as a colorless oil, which was used directly in the next step without further purification. MS measured value (ESI + )[(M+H) + ]:346.4.

[0247] Step 8: (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]morpholine-4-carboxamide (Example 29) [ka] Example 29 was prepared in the same manner as the preparation of compound Example 20, by using 29 g instead of tert-butyl N-carbamimidylcarbamate and DIEA instead of K2CO3 in step 2. Example 29 (26.1 mg, 0.03 mmol, yield 56.3%) was obtained as a white solid in the form of hydrochloride. MS measurement (ESI + )[({ 35 Cl}M+H) + ]:754.5.

[0248] Example 24: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(3-aminopropyl)guanidine [ka] Step 1: tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(imidazole-1-carboxyimidoylamino)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of Int-2 (60.0 mg, 0.1 mmol) in DMF (0.4 mL), imidazole-1-yl (1,2,4-triazole-4-yl)methanymine (85.4 mg, 0.5 mmol) was added under N2 at 25°C, and the mixture was stirred at 45°C for 12 hours. After cooling to room temperature, the mixture was purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 12 min), and lyophilized to obtain 24a (50.0 mg, 0.1 mmol, yield 34.5%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:776.3.

[0249] Step 2: tert-butyl N-[3-[[N-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamimidoyl]amino]propyl]carbamate [ka] To a solution of 24a (50.0 mg, 0.1 mmol) in DMF (0.2 mL), NB℃-1,3-diaminopropane (157.1 mg, 0.9 mmol) was added under N2 at 25°C, and the mixture was heated to 80°C for 3 hours with stirring. After cooling to room temperature, the mixture was purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain 24b (30.0 mg, 0.03 mmol, yield 37.7%) as a yellow solid. MS measured value (ESI + )[{ 35 Cl}M+H] + ]:882.5.

[0250] Step 3: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(3-aminopropyl)guanidine [ka] To a solution of 24b (25.0 mg, 0.03 mmol) in DCM (1 mL), TFA (1 mL) was added at 25°C and the mixture was stirred at 25°C for 1 hour. After concentrating the mixture, the residue was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 14%~44%, 10 min), lyophilized, and 24 (5.6 mg, 0.01 mmol, yield 25.7%) was obtained as a yellow solid in the form of its hydrochloride salt. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:682.3.

[0251] Example 25: N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]-2,2,2-trifluoroacetamide [ka] Step 1: tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-[[(2,2,2-trifluoroacetyl)amino]methyl]cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of Int-4 (70.0 mg, 0.1 mmol) in DCM (0.7 mL), trifluoroacetic anhydride (63.0 mg, 0.3 mmol) and DIEA (129.5 mg, 1.0 mmol) were added at 25°C. The mixture was stirred at 25°C for 2 hours, then quenched with water (30 mL) and extracted with ethyl acetate (60 mL x 3). The combined organic layer was washed with saturated brine (60 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum to obtain the residue, which was then preparatively collected as TLC (PE / ethyl acetate = 1 / 2; TLC: PE / ethyl acetate = 1 / 2, R f Purified by (=0.5), 25a (50.0 mg, 0.1 mmol, yield 62.8%) was obtained as a pale yellow solid. MS measured value (ESI+)([{ 35 Cl}M+H] + ):793.3.

[0252] Step 2: N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]-2,2,2-trifluoroacetamide [ka] To a solution of 25a (35.0 mg, 0.04 mmol) in DCM (1 mL), TFA (2.0 mL) was added at 25°C and stirred for 1 hour at 25°C. After concentrating the mixture under vacuum, the residue was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100 × 25 mm × 4 μm; mobile phase: [A: water (0.1% TFA)-ACN]; B%: 20%~60%, 10 min), lyophilized, and 25 (9.2 mg, 0.01 mmol, yield 24.0%) was obtained as a white solid in the form of TFA salt. MS measured value (ESI+) ([{ 35 Cl}M+H] + ):693.2.

[0253] Example 36: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] Step 1: Di-tert-butyl(((2-((tert-butyldimethylsilyl)oxy)ethyl)azandiyl)bis(ethane-2,1-diyl))dicarbamate [ka] To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (7.2 g, 23.7 mmol) in acetonitrile (30 mL), (2-bromoethoxy)-tert-butyldimethylsilane (5.7 g, 23.7 mmol) and NaHCO3 (6.0 g, 71.2 mmol) were added at 25 °C. The mixture was stirred at 45 °C for 72 hours, then diluted with siRNA (100 mL) and filtered. The filtrate was concentrated under vacuum to obtain a residue, which was purified by silica gel (siRNA / MeOH = 1:0~10 / 1; TLC: siRNA / MeOH = 10 / 1, Rf = 0.3) to obtain 36a (2.7 g, 5.9 mmol, yield 24.6%) as a colorless oil. (ESI+)[(M+H)+]:462.1.

[0254] Step 2: Di-tert-butyl(((2-hydroxyethyl)azandiyl)bis(ethane-2,1-diyl))dicarbamate [ka] To a solution of 36a (6.6 g, 14.3 mmol) in water (7.0 mL) and THF (30 mL), AcOH (13 mL) was added at 20°C and the mixture was stirred for 12 hours. The mixture was purified by preparative HPLC (column: C18 spherical 20-35 μm, 330 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0%-100%, 25 min), and lyophilized to obtain 36b (2.7 g, 7.8 mmol, yield 54.4%) as a colorless oil. MS measurement value (ESI+)[(M+H)+]: 348.3.

[0255] Step 3: 2-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]ethyl(4-nitrophenyl) carbonate [ka] To a solution of 4-nitrophenyl chloroformate (319.1 mg, 1.6 mmol) and 36b (500.0 mg, 1.4 mmol) in THF (15 mL), N,N-diisopropylethylamine (558.0 mg, 4.3 mmol) was added at 25 °C, and the mixture was heated to 40 °C with stirring for 48 hours. The mixture was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain 36c (270.0 mg, 0.5 mmol, yield 36.6%) as a colorless oil. MS measured value (ESI+)[(M+H)] + ]:513.3.

[0256] Step 4: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]ethoxycarbonylamino]cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of Int.2 (200.0 mg, 0.3 mmol) and 36c (300.1 mg, 0.6 mmol) in THF (15 mL), N,N-diisopropylethylamine (0.5 mL, 2.9 mmol) was added at 25 °C, and the mixture was heated to 60 °C with stirring for 48 hours. After concentrating the mixture under vacuum, the residue was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% TFA)-B: ACN]; B%: 0%-100%, 15 min), and lyophilized to obtain 36d (80.0 mg, 0.1 mmol, yield 25.9%) as a white solid. (ESI + )[({ 35 Cl}M+H) + ]:1056.7.

[0257] Step 5: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] To a solution of 36d (60.0 mg, 0.1 mmol) in xDCM (2 mL), TFA (1 mL) was added at 25°C and stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 14%~44%, 10 min), lyophilized, and 36 (17.8 mg, 0.02 mmol, yield 40.2%) was obtained as a white solid in the form of its hydrochloride salt. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:756.1.

[0258] Example 27: 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] Example 27 was prepared in the same manner as the preparation of compound 36, except that in step 1, di-tert-butyl(azandiylbis(ethane-2,1-diyl))dicarbamate was replaced with di-tert-butyl(azandiylbis(propane-3,1-diyl))dicarbamate. Example 27 (6.3 mg, 0.01 mmol, yield 43.0%) was obtained as a white powder in the form of a TFA salt. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:784.4.

[0259] Example 28: 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)-(carboxymethyl)ammonium [ka] Step 1: tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propyl-[2-[tert-butyl(dimethyl)silyl]oxyethyl]amino]propyl]carbamate [ka] To a solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (1.0 g, 3.0 mmol) in acetonitrile (10 mL), (2-bromoethoxy)-tert-butyldimethylsilane (721.8 mg, 3.0 mmol) and NaHCO3 (760.4 mg, 9.1 mmol) were added at 45°C. The mixture was stirred at 45°C for 72 hours, then filtered, and the filtrate was concentrated under vacuum to obtain the residue. This residue was purified by silica gel column chromatography (siRNA / MeOH=1 / 0~10 / 1; TLC: siRNA / MeOH=10 / 1, Rf=0.3) to obtain 28a (950.0 mg, 1.9 mmol, yield 64.3%) as a colorless oil. MS measured value (ESI + )[(M+H) + ]:490.4.

[0260] Step 2: 2-[bis(3-aminopropyl)amino]ethanol [ka] A mixture of 28a (500.0 mg, 1.0 mmol) in HCl / MeOH (4.0 M, 10.0 mL) was stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain crude 28b (216.0 mg, 1.0 mmol, yield 99.9%) as a colorless oil, which was used directly in the next step without further purification. MS measured value (ESI + )[(M+H) + ]:176.0

[0261] Step 3: tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propyl-(2-hydroxyethyl)amino]propyl]carbamate [ka] To a solution of 28b (216.0 mg, 1.0 mmol) in methanol (15 mL), sodium bicarbonate (342.8 mg, 4.1 mmol) and di-t-butyl dicarbonate (445.3 mg, 2.0 mmol) were added at 25°C. The mixture was stirred at 25°C for 1 hour, then diluted with Depositphotos (50 mL) and filtered. The filtrate was concentrated under vacuum to obtain the residue, which was then filtered through a silica gel column ( Depositphotos / MeOH = 1 / 0 to 10 / 1; TLC: Depositphotos / MeOH = 10 / 1, R f Purified by (=0.3), 28c (180.0 mg, 0.5 mmol, yield 47.0%) was obtained as a yellow oily substance. MS measured value (ESI + )[(M+H) + ]:376.1.

[0262] Step 4: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-[bis[3-(tert-butoxycarbonylamino)propyl]amino]ethoxycarbonylamino]cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of 28c (50.0 mg, 0.1 mmol) in THF (2 mL), N,N-diisopropylethylamine (0.1 mL, 0.6 mmol) and 4-nitrophenyl chloroformate (30.0 mg, 0.2 mmol) were added under N2 conditions at 0°C. After stirring at 40°C for 48 hours, Int-2 (100.1 mg, 0.2 mmol) and 4-dimethylaminopyridine (5.0 mg, 0.04 mmol) were added, and the mixture was stirred at 40°C for 48 hours. The mixture was concentrated under vacuum to obtain the residue. The residue was dissolved in DMF (2 mL), then purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 10 min), and lyophilized to obtain 28d (50.0 mg, 0.1 mmol, yield 34.6%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:1084.7.

[0263] Step 5: N-(2-(tert-butoxy)-2-oxoethyl)-3-((tert-butoxycarbonyl)amino)-N-(2-((((1r,4r)-4-((2-(4-((4-((R)-4-((tert-butoxycarbonyl)amino)-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)carbamoyl)oxy)ethyl)-N-(3-((tert-butoxycarbonyl)amino)propyl)propane-1-aminium [ka] A mixture of 28d (45.0 mg, 0.04 mmol), tert-butylbromoacetate (24.3 mg, 0.1 mmol), and DIEA (0.2 mL, 1.2 mmol) in DMF (0.2 mL) was stirred at 70°C for 3 hours. The mixture was acidified to pH=5 with FA, followed by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% FA)-B: ACN]; B%: 0%-100%, 10 min), and lyophilized to obtain 28e (50.0 mg, 0.04 mmol, yield 87.8%) as a yellow solid. MS measured values ​​(ESI + )[({ 35 Cl}M+H) + ]:1198.5

[0264] Step 6: 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)-(carboxymethyl)ammonium (Example 28) [ka] To a solution of 28e (40.0 mg, 0.03 mmol) in DCM (2 mL), TFA (2 mL) was added under N2 at 25°C. After stirring the mixture at 25°C for 1 hour, it was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15%~45%, 10 min), lyophilized, and 28 (15.0 mg, 0.02 mmol, yield 47.9%) was obtained as a brown solid in the form of its hydrochloride salt. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:842.3.

[0265] Example 30: N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide [ka] Step 1: tert-butyl N-[4-[1-(2,6-dichloro-4-pyridyl)vinyl]cyclohexyl]carbamate [ka] To a mixture of methyltriphenylphosphonium bromide (7.5 g, 20.9 mmol) in THF (70 mL), n-butyllithium solution (2.5 M, 7.5 mL, 18.8 mmol) was added dropwise at 0°C. Int-2c (3.9 g, 10.5 mmol) in THF (70 mL) solution was then added, and the mixture was stirred at 0°C for 2 hours. The reaction mixture was poured into 100 mL of saturated NH4Cl aqueous solution and extracted with Depositphotos (100 mL x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to obtain the residue, which was then filtered through a silica gel column (PE / Depositphotos=50 / 1~2 / 1; TLC:PE / Depositphotos=2 / 1, R f Purified by (=0.4), 30a (2.9g, 7.8 mmol, yield 74.8%) was obtained as a yellow solid. MS measured value (ESI + )[({ 35 Cl}M-C4H8+H) + ]:315.0.

[0266] Step 2: tert-butyl N-[4-[1-(2,6-dichloro-4-pyridyl)cyclopropyl]cyclohexyl]carbamate [ka] Trimethylsulfoxonium iodide (200.0 mg, 0.9 mmol) was dissolved in DMSO (2 mL), and sodium hydride (38.0 mg, 60% wt., 1.1 mmol) was added all at once at 0°C. Then, 30a (200.0 mg, 0.5 mmol) was added to the solution at 0°C, and the mixture was stirred continuously at 25°C for 2 hours. The mixture was poured into 20 mL of saturated NH4Cl aqueous solution and extracted with  (50 mL x 3). The organic layers were combined, washed with saturated brine (50 mL x 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under vacuum to obtain the residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 30b (160.0 mg, 0.4 mmol, yield 77.1%) was obtained as a yellow solid. MS measured value (ESI+)[({ 35 Cl}M-C4H8+H) + ]:329.0.

[0267] Step 3: tert-butyl N-[4-[1-(2,6-dichloro-4-pyridyl)cyclopropyl]cyclohexyl]carbamate [ka] To a solution of 30b (200.0 mg, 0.5 mmol) in DCM (2 mL), TFA (2 mL) was added all at once at 25°C and stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain crude 30c (160.0 mg, 0.6 mmol) as a white solid, which was used directly in the next step without further purification. MS measured value (ESI+): 295.2[({ 35 Cl}M+H)+].

[0268] Step 4: tert-butyl N-(4-formylcyclohexyl)carbamate [ka] To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (500.0 mg, 1.7 mmol) and benzyl 4-bromobutanoate (423.7 mg, 1.7 mmol) in ACN (10 mL), sodium bicarbonate (1.4 g, 16.5 mmol) was added all at once at 25°C. The mixture was stirred at 25°C for 15 hours, then diluted with SiO2 (200 mL) and filtered. The filtrate was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 30d (380.0 mg, 0.8 mmol, yield 48.1%) as a yellow oil. MS measured value (ESI+)[(M+H)] + ]:480.2.

[0269] Step 5: 4-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]butanoic acid [ka] To a solution of 30d (180.0 mg, 0.4 mmol) in methanol (20 mL), wet Pd / C (50 mg, 5% wt, containing 50% water) was added under an N2 atmosphere at 20°C. The suspension was degassed under reduced pressure and purged three times with H2. The reaction mixture was then stirred under H2 (15 psi) at 40°C for 16 hours. The mixture was diluted with methanol (200 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 10 min), lyophilized, and 30e (120.0 mg, 0.3 mmol, yield 82.1%) as a colorless oil. MS measured value (ESI+)[(M+H)] + ]:390.2.

[0270] Step 6: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[1,4-dioxaspiro[4.5]decane-8-yl(difluoro)methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of 30e (98.3 mg, 0.3 mmol) and 30b (60.0 mg, 0.2 mmol) in DMF (2 mL), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (96.0 mg, 0.3 mmol) and N,N-diisopropylethylamine (742.0 mg, 5.7 mmol) were added in one step at 0°C. The mixture was stirred at 25°C for 2 hours, then concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 40 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 10 min), lyophilized, and 30f (60.0 mg, 0.1 mmol, yield 43.4%) as a yellow solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:656.3.

[0271] Step 7: tert-butyl N-[(3R)-1-[4-[4-[4-[1-[4-[4-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]butanoylamino]cyclohexyl]cyclopropyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of 30f (200.0 mg, 0.3 mmol) in DMSO (1 mL), Int-1 (129.3 mg, 0.3 mmol) and N,N-diisopropylethylamine (116.0 mg, 0.9 mmol) were added at 25°C, and the mixture was heated to 100°C with stirring for 16 hours. After cooling to 25°C, the mixture was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: C18 spherical 20-35 μm, 100 g, mobile phase: [A: water (0.1% FA) - B: ACN]; B%: 0%-100%, 15 min), lyophilized, and 30 g (80.0 mg, 0.1 mmol, yield 25.1%) was obtained as a yellow solid. MS measured value (ESI + )[({ 35 Cl}MB℃+H) + ]:944.4.

[0272] Step 8: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of 30 g (60.0 mg, 0.1 mmol) in DCM (2 mL), TFA (3 mL) was added all at once at 25°C, and the mixture was stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain a residue, which was then subjected to preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15%~45%, 10 min), and lyophilized to obtain 30 (8.0 mg, 0.01 mmol, yield 17.4%) as a white solid in the form of hydrochloride. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:744.3.

[0273] Example 33: (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxypropyl)amino]cyclohexyl]-difluoromethyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Step 1: 3-[tert-butyl(dimethyl)silyl]oxy-2,2-difluoropropane-1-ol [ka] To a solution of 2,2-difluoropropane-1,3-diol (1.0 g, 8.9 mmol) in DMF (20 mL), imidazole (1.2 g, 17.8 mmol) and tert-butyldimethylchlorosilane (1.6 g, 10.8 mmol) were added at 0°C. The mixture was stirred at 0°C for 4 hours, then diluted with H2O (30 mL) and transferred to a separatory funnel. The aqueous layer was extracted with Depositphotos (80 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was then filtered through a silica gel column (PE / Depositphotos=1 / 0~5 / 1; TLC:PE / Depositphotos=5 / 1, R f The solution was purified using a method (=0.4) to obtain 33a (2.0 g, 98.8% yield) as a colorless oil. 33a was used directly in the next step without any characterization analysis.

[0274] Step 2: 3-[tert-butyl(dimethyl)silyl]oxy-2,2-difluoropropanal [ka] To a solution of 33a (300.0 mg, 1.3 mmol) in DCM (5 mL), DMP (618.4 mg, 1.5 mmol) was added at 0°C and stirred for 2 hours. Slowly add saturated K2CO3 solution (20 mL) to quench the mixture, then transfer to a separatory funnel and extract the aqueous layer with  (40 mL × 3). The combined organic layers were washed with saturated brine (50 mL × 2), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was then analyzed by silica gel column (PE / Â=1 / 0~5 / 1; TLC:PE / Â=5 / 1, R f The solution was purified using a method (=0.5) to obtain 33b (250.0 mg, 1.1 mmol, yield 84.1%) as a colorless oil. 33b was used directly in the next step without characterization analysis.

[0275] Step 3: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[[3-[tert-butyl(dimethyl)silyl]oxy-2,2-difluoro-propyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]-5-oxo-pyrrolidine-3-yl]carbamate [ka] To a solution of 33b (60.0 mg, 0.2 mmol) and Int-2 (201.0 mg, 0.3 mmol) in DCE (1 mL), acetic acid (32.1 mg, 0.5 mmol) and sodium triacetoxyborohydride (283.4 mg, 1.3 mmol) were added in one step under N2 at 25°C. The mixture was stirred at 25°C for 16 hours, and then diluted with  (100 mL) and water (20 mL). The organic layer was separated, washed with saturated brine (30 mL x 2), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain crude 33c (50.0 mg, 0.01 mmol, yield 48.4%) as a white solid, which was used directly in the next step without further purification. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:891.2.

[0276] Step 4: (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxypropyl)amino]cyclohexyl]-difluoromethyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] To a solution of 33c (30.0 mg, 0.03 mmol) in DCM (1 mL), TFA (1 mL) was added at 25°C and stirred at 25°C for 2 hours. The mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 20%~50%, 10 min), lyophilized, and 33 (7.5 mg, 0.01 mmol, yield 28.4%) as a white solid. MS measured value (ESI + )[({ 35 Cl}M+H) + ]:677.2.

[0277] Example 37: (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-methylpyrimidine-2-yl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Step 1: (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoromethyl]-6-methylpyrimidine-2-yl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one [ka] Int-5j (35.0 mg, 0.04 mmol) was added to TFA (2.0 mL, 26.0 mmol) at 25°C and heated to 60°C with stirring for 16 hours. The mixture was concentrated under reduced pressure to obtain a residue, which was then subjected to preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 12%~42%, 10 min). Lyophilization yielded 37 (19.4 mg, 0.03 mmol, yield 75.6%) as a white solid in the form of hydrochloride. MS measured value (ESI + )[(M+H) + ]:564.4.

[0278] Example 38: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-methylpyrimidine-4-yl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide [ka] Example 38 was prepared in the same manner as the preparation of Compound Example 15, by using 32b instead of 2-[2-[(2-methylpropan-2-yl)oxycarbonylamino]ethoxy]acetic acid and Int-5 instead of Int-4 in step 1. Example 38 (3.5 mg, 0.01 mmol, yield 7.3%) was obtained as a white solid in the form of the hydrochloride salt. MS measurement (ESI + )[(M+H) + ]:763.6.

[0279] Example 40: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate [ka] Step 1: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; trihydrochloride [ka] To a solution of Int-7 (50.0 mg, 0.1 mmol) in DCM (2 mL), trifluoroacetic acid (1.00 mL, 12.98 mmol) was added at 25°C and the mixture was stirred for 1 hour. The mixture was concentrated under vacuum to obtain a residue, which was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15%~45%, 10 min), lyophilized, and Example 40 (32.4 mg, 0.1 mmol, yield 70.7%) was obtained as a white solid in the form of hydrochloride. MS measured value. (ESI) + )[({ 35 Cl}M+H) + ]:597.3.

[0280] Example 39: (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one [ka] Step 1: tert-butyl((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] To a solution of Int-5 (500.0 mg, 0.8 mmol) in isopropanol (2 mL), 1,1,1-trifluoro-2,3-epoxypropane (84.4 mg, 0.8 mmol) and DIEA (294.1 mg, 2.3 mmol) were added at 25°C, and the mixture was heated to 70°C for 2 hours with stirring. The mixture was quenched by adding saturated ammonium chloride aqueous solution (20 mL), then transferred to a separatory funnel, and the aqueous layer was extracted with ethyl acetate (60 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product, which was then filtered through a silica gel column (ethyl acetate / MeOH = 1 / 0 to 10 / 1, with 1‰NH₃·H₂O as an additive; TLC: ethyl acetate / MeOH = 10 / 1R). f Purified by (=0.4), 39a (400.0 mg, 0.5 mmol, yield 67.8%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:776.2

[0281] Step 2: (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one [ka] To a solution of 39a (400.0 mg, 0.5 mmol) in DCM (5 mL), TFA (3 mL) was added at 25°C, and the mixture was stirred at 25°C for 2 hours. After concentrating the mixture under reduced pressure, crude Example 39 (304.0 mg, 0.5 mmol, yield 88.0%) was obtained as a white solid. MS measured value (ESI + )[(M+H) + ]:676.2.

[0282] Example 42: tert-butyl((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate [ka] Step 1: tert-butyl((1r,4S)-4-((2-(4-((4-((R)-4-((tert-butoxycarbonyl)amino)-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazin-1-yl)-6-methylpyrimidine-4-yl)difluoromethyl)cyclohexyl)((S)-3,3,3-trifluoro-2-hydroxypropyl)carbamate(42a ) and tert-butyl((1r,4R)-4-((2-(4-((4-((R)-4-((tert-butoxycarbonyl)amino)-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-methylpyrimidine-4-yl)difluoromethyl)cyclohexyl)((R)-3,3,3-trifluoro-2-hydroxypropyl)carbamate(42b) [ka] [ka] To a solution of 39 (300.0 mg, 0.4 mmol) in DCM (5 mL), di-t-butyl dicarbonate (210.0 mg, 1.0 mmol) and DIEA (148.9 mg, 1.2 mmol) were added at 25°C, and the mixture was stirred at 25°C for 2 hours. The resulting mixture was poured into water (50 mL), transferred to a separatory funnel, and the aqueous layer was extracted with ethyl acetate (80 mL x 3). The combined organic layers were washed with brine (50 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product, which was then analyzed on a silica gel column (PE / ethyl acetate = 10 / 1 to 0 / 1; TLC: PE / ethyl acetate = 1 / 1, R). fThe sample was purified by (=0.4), and then separated by preparative SFC (column: Chiralpak IG 25×30mm ID, 10um, mobile phase: 40% IPA (0.1% NH3·H2O) in supercritical CO2), yielding 42a (first to elute, 140.0 mg, 0.16 mmol, yield 41.54%) as a white solid. MS measured value (ESI + )[(M+H) + ]:876.2 and 42b (second to elute, 160.0 mg, 0.18 mmol, yield 47.47%) were obtained as white solids. MS measured values ​​(ESI + )[(M+H) + ]:876.2.

[0283] Step 2: (R)-4-amino-1-(4-((4-(4-(difluoro((1R,4r)-4-(((R)-3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one [ka] To a solution of 42b (120.0 mg, 0.1 mmol) in DCM (2 mL), TFA (2 mL) was added at 25°C and the mixture was stirred at 25°C for 2 hours. After concentrating the reaction under reduced pressure, the residue was purified by preparative HPLC (column: YMC Triart C18 150 × 25 mm × 5 μm, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 18%~48%, 10 min), lyophilized, and compound 42 (71.2 mg, 0.1 mmol, yield 73.6%) was obtained as a white solid in the form of its hydrochloride salt. MS measurement values ​​(ESI + )[(M+H) + ]:676.3.

[0284] Example 43: Minimum Inhibitory Concentration Protocol (MIC) Assay: The antibacterial activity of the compounds of the present invention was evaluated against the commonly used quality control strain of Escherichia coli (ATCC 25922) and the rifampin-resistant mutant strain of Klebsiella pneumonia (ATCC 43816). Both strains are originally derived from human clinical samples and are available from the ATCC (American Type Culture Collection). The in vitro efficacy of the compounds that inhibit the growth of Escherichia coli (ATCC 25922) and Klebsiella pneumonia (ATCC 43816) was evaluated by the MIC (minimum inhibitory concentration) broth dilution method. Specifically, compound dilutions were prepared from 10 mM DMSO stock solution as follows: i) A series of 2-fold dilutions in 20 μL DMSO were prepared on a master plate (Greiner, catalog number: 651201), ii) 180 μL of sterile distilled water was added to each aliquot, and iii) 10 μL of the diluted compound was transferred to a new assay plate (Costar, 3599).

[0285] The vials of each test microorganism were kept frozen in the gas phase of a liquid nitrogen freezer. Single-use frozen vials of two strains, Escherichia coli ATCC 25922 (KWIKSTIK, 0335K) and Klebsiella pneumonia ATCC 43816, with predetermined CFU / mL levels, were removed from the freezer, thawed at room temperature, and diluted with cation-modified Mueller-Hinton broth (CAMHB) to 5 × 10⁻⁶. 5 The final inoculation dose of CFU / mL was achieved. 90 μL of broth containing bacteria was dispensed into an assay plate containing pre-dispensed compound dilutions and mixed five times by pipetting.

[0286] Next, the assay plate was incubated in ambient air at 35°C for 20 hours. After incubation, the minimum MIC (μg / mL) of the drug that inhibited visible microbial growth was read and recorded with the help of a magnifying glass.

[0287] Compound F-1, disclosed in the literature (Bioorganic Chemistry (102), 2020, 104055) and reported to have LPS synthesis pathway inhibitory activity and antibiotic activity against efflux-deficient Escherichia coli strains, was selected as the reference compound in this invention. [ka]

[0288] [Table 1]

[0289] Example A Film-coated tablets containing the following ingredients can be manufactured in the conventional manner. [Table 2]

[0290] The active ingredients are sieved and mixed with microcrystalline cellulose, and the mixture is granulated with an aqueous solution of polyvinylpyrrolidone. The granules are then mixed with sodium starch glycolate and magnesium stearate and compressed to obtain 120 or 350 mg of nuclei, respectively. The nuclei are lacquered with an aqueous solution / suspension of the above film coat.

[0291] Example B Capsules containing the following ingredients can be manufactured in a conventional manner. [Table 3]

[0292] The ingredients are sifted, mixed, and filled into size 2 capsules.

[0293] Example C The injection solution may have the following composition: [Table 4]

[0294] The active ingredient is dissolved in a mixture of polyethylene glycol 400 and part of sterile water for injection. The pH is adjusted to 5.0 by adding acetic acid. The remaining water is added to adjust the volume to 1.0 ml. The solution is filtered, and the appropriate excess volume is used to fill the vial, which is then sterilized.

Claims

1. Equation (I) 【Chemistry 1】 (In the formula, L is either absent or alkyl. X is -O- or -NH-, n is either 0 or 1. R1 is amino, aminoalkyl, alkylamino, aminoalkylamino, haloalkyl, diaminoalkyl, guanidinyl, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkyl, aminoalkoxyalkyl, (aminoalkyl)(aminoalkyl)(aminocarbonylalkyl)ammoniumalkyl, (amino)(hydroxy)alkyl, (aminoalkyl)(aminoalkyl)amino, (aminoalkyl)(aminoalkyl)aminoalkyl, (aminoalkyl)(aminoalkyl)aminoalkoxy, (aminoalkyl)(aminoalkyl)aminoalkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonyl)ammoniumalkylamino, (hydroxy)(halo)alkylamino, (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkoxy, cycloalkyl, or heterocycloalkyl, where heterocycloalkyl and cycloalkyl are optionally substituted with one or two substituents independently selected from R6. However, if R1 is an amino acid, L does not exist. R2 and R3 are independently selected from halogens, or R2 and R3 together with the carbon to which they are bonded form a cycloalkyl group. R4 is a halogen or alkyl group. R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino, or piperazinyl. In each example, R6 is independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl, and aminoalkylcarbonylamino. R7 is hydrogen or hydroxyl, A １ is -N- or -CH-, A ２ is -O- or -CH ２ - is a compound of or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein L is absent.

3. The compound according to claim 1 or 2, wherein L is alkyl, particularly methyl.

4. A compound according to any one of claims 1 to 3, wherein n is 1.

5. The compound according to any one of claims 1 to 4, wherein X is -O-.

6. A compound according to any one of claims 1 to 3, wherein n is 0.

7. R1 is amino, aminomethyl, aminoethyl, methylamino, aminoethylamino, aminopropylamino, trifluoromethyl, diaminoethyl, guanidinyl, aminoethoxymethyl, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropyl, (aminopropyl)(aminopropyl)(aminocarbonylmethyl)ammoniumpropyl, (amino)(hydroxy)ethyl, (aminopropyl)(aminopropyl)amino, (aminoethyl)(aminoethyl)aminopropyl, (aminopropyl)(aminopropyl)aminopropyl, (aminopropyl)(aminopropyl)aminoethoxy, (aminoethyl)(aminoethyl)aminoethoxy, (aminoethyl)(aminoethyl)ami Noethylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropylamino, (hydroxy)difluoropropylamino, (hydroxy)trifluoropropylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumethoxy, cyclobutyl, azetidine-3-yl, azetidine-1-yl, morpholine-4-yl, pyrrolidine-1-yl, or 5-azoniaspiro[2.4]heptane, wherein cyclobutyl, azetidine-3-yl, azetidine-1-yl, morpholine-4-yl, pyrrolidine-1-yl, or 5-azoniaspiro[2.4]heptane is optionally substituted with one or two substituents independently selected from R6. However, if R1 is an amino acid, L is absent, the compound according to any one of claims 1 to 6.

8. The compound according to any one of claims 1 to 6, wherein R1 is a heterocycloalkyl group in which R1 is optionally substituted with one or two substituents independently selected from R6.

9. The compound according to any one of claims 1 to 8, wherein both R2 and R3 are fluorides, or R2 and R3, together with the carbons to which they are bonded, form a cyclopropyl group.

10. The compound according to any one of claims 1 to 9, wherein R4 is chloride or methyl.

11. The compound according to any one of claims 1 to 10, wherein R5 is an amino or hydroxyalkyl, particularly an amino.

12. The compound according to any one of claims 1 to 11, wherein R6 is independently selected from amino, methyl, hydroxy, aminomethyl, aminocarbonylmethyl, and aminoethylcarbonylamino in each example.

13. The compound according to any one of claims 1 to 12, wherein R7 is hydrogen.

14. A １ The compound according to any one of claims 1 to 13, wherein is -CH-.

15. A ２ ga-CH ２ - The compound according to any one of claims 1 to 14.

16. below, cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidine-1-yl)cyclohexyl]methyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]azetidine-3-yl]propanamide; (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; trans-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]propanamide; trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]cyclobutanecarboxamide; cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-hydroxypropanamide; cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]urea; trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-5,5-dimethyl-5-azoniaspiro[2.4]heptan-6-carboxamide; trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptan-6-carboxamide; trans-(5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptan-6-carboxamide; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]acetamide; 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N-(carboxymethyl)-4-oxobutan-1-aminium; N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-(2-aminoethoxy)acetamide; 3-amino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-2-hydroxypropanamide; 2,3-diamino-N-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)propanamide; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(diaminomethylene)urea; (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-4-oxo-butyl]-bis(3-aminopropyl)ammonium; 3-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoylamino]propyl-bis(3-aminopropyl)-(carboxymethyl)ammonium; 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-3-(3-aminopropyl)guanidine; N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]methyl]-2,2,2-trifluoroacetamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)-(carboxymethyl)ammonium; (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]morpholine-4-carboxamide; N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N-hydroxybutanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxypropyl)amino]cyclohexyl]-difluoromethyl]-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]-4-hydroxypyrrolidine-1-carboxamide; 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidine-1-yl)phenyl)sulfonyl)piperazine-1-yl)-6-chloropyridine-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2-aminoethyl)amino)ethyl)urea; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-methylpyrimidine-2-yl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-methylpyrimidine-4-yl]-difluoromethyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)pyrrolidine-2-one; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxopyrrolidine-1-yl]phenyl]sulfonylpiperazine-1-yl]-6-chloro-4-pyridyl]-difluoromethyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[[4-(azetidine-1-yl)cyclohexyl]-difluoromethyl]-6-chloro-2-pyridyl]piperazine-1-yl]sulfonylphenyl]pyrrolidine-2-one; tert-butyl((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2-hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidine-2-yl)piperazine-1-yl)sulfonyl)phenyl)-5-oxopyrrolidine-3-yl)carbamate; A compound selected from, or a pharmaceutically acceptable salt thereof.

17. A compound according to any one of claims 1 to 16, for use as a therapeutically active substance.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 and a therapeutically inert carrier.

19. Use of the compound according to any one of claims 1 to 16 for the treatment or prevention of bacterial infections, particularly bacterial infections in which the bacteria are Gram-negative bacteria.

20. Use of a compound according to any one of claims 1 to 16 for the preparation of a pharmaceutical product for the treatment or prevention of a bacterial infection, particularly a bacterial infection in which the bacteria are Gram-negative bacteria.

21. A compound according to any one of claims 1 to 16 for use in the treatment or prevention of bacterial infections, particularly bacterial infections in which the bacteria are Gram-negative bacteria.

22. A method for treating or preventing a bacterial infection, particularly an infection caused by Gram-negative bacteria, comprising administering an effective amount of any one of claims 1 to 16.

23. The use, method, or compound for use according to any one of claims 19 to 22, wherein the Gram-negative bacteria are selected from Enterobacteriaceae, Neisseria gonorrhoeae, Haemophilus influenzae, Helicobacter pylori, Acinetobacter baumannii, and Pseudomonas aeruginosa.

24. The use, method, or compound for use of the compound according to claim 23, wherein the Gram-negative bacterium is of the Enterobacteriaceae family, and the Enterobacteriaceae family is Klebsiella pneumoniae or Escherichia coli.

25. The invention as previously described herein.

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