EP4695237A1

EP4695237A1 - Novel compounds for the treatment of bacterial infections

Info

Publication number: EP4695237A1
Application number: EP24720039.7A
Authority: EP
Inventors: Bjoern Bartels; Fabian Dey; Xiao DING; Satoshi Hashimoto; Christian Lerner; Houguang SHI; Takao Suzuki; Xuefei Tan; Jun Wu; Jiamin Zheng
Original assignee: F Hoffmann La Roche AG
Current assignee: F Hoffmann La Roche AG
Priority date: 2023-04-12
Filing date: 2024-04-11
Publication date: 2026-02-18
Also published as: WO2024213625A1; JP2026514062A; CN121127463A

Abstract

The invention relates to a compound of formula (I), wherein R1-R7, n, X, L, A₁ and A₂ are as defined in the description and in the claims. The compound of formula (I) can be used as a medicament.

Description

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Novel compounds for the treatment of bacterial infections The present invention relates to organic compounds useful for the treatment and/or prevention of bacterial infections in a mammal. Specifically these molecules can inhibit the LPS synthesis pathway, in particular to inhibit LpxH, and are useful for treating bacterial infections. The invention relates in particular to a compound of formula (I) wherein L is absent or alkyl; X is -O- or -NH-; n is 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, F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (aminoalkyl)(aminoalkyl)(hydroxycarbonylalkyl)ammoniumalkoxy, cycloalkyl or heterocyloalkyl, wherein heterocycloalkyl and cycloalkyl are optionally substituted by one or two substituents independently selected from R6; with the proviso that if R1 is amino, then L is absent; R2 and R3 are independently selected from halogen; or R2 and R3 together with the carbon they are attached to form cycloalkyl; R4 is halogen or alkyl; R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino or piperazinyl; R6 is at each instance independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl and aminoalkylcarbonylamino; R7 is hydrogen or hydroxy; A1 is -N- or -CH-; and A₂ is -O- or -CH₂-; or a pharmaceutically acceptable salt thereof. The intensive use of antibiotics has exerted a selective evolutionary pressure on microorganisms to produce genetically based resistance mechanisms. Modern medicine and socio- economic behaviour exacerbate the problem of resistance development by creating slow growth situations for pathogenic microbes, e.g. in artificial joints, and by supporting long-term host reservoirs, e.g. in immune-compromised patients. In hospital settings, an increasing number of strains of Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus spp., Enterobacteriaceae such as Klebsiella pneumonia, Acinetobacter baumannii and Pseudomonas aeruginosa, major sources of infections, are becoming multi-drug resistant and therefore difficult to treat. This is particularly the case for Gram-negative organisms where the situation is getting worrisome since no novel agents with a differentiated mechanism of action have been approved for decades. Therefore, there is an important medical need for new antibacterial compounds addressing Gram-negative resistant bacteria, in particular third generation cephalosporins- and carbapenem-resistant Enterobacteriaceae and multi-drug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii. One way to tackle the problem of cross-resistance to established classes of antibiotics is to inhibit an essential protein or function not targeted by current antibiotics. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Gram-negative bacteria are unique in that their outer membrane contains Lipopolysaccharide (LPS), which is crucial for maintaining membrane integrity, and is essential for bacterial viability (reviewed in Ann. Rev. Biochem 76: 295-329, 2007). The major 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 that consists 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 yield Lipid X and UMP. LpxH has no mammalian homologue, making it a good target for the development of novel antibiotics targeting Gram-negative bacteria. The term "alkyl", alone or in combination, denotes a monovalent linear or branched saturated hydrocarbon group of 1 to 12 carbon atoms, in particular of 1 to 7 carbon atoms, more particular of 1 to 4 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, n - butyl, iso-butyl, sec-butyl, or tert-butyl. Methyl, ethyl and propyl are particular examples of “alkyl”. The term “cycloalkyl”, alone or in combination, signifies a cycloalkyl ring with 3 to 8 carbon atoms and particularly a cycloalkyl ring with 3 to 6 carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Particular examples of “cycloalkyl” are cyclopropyl and cyclobutyl. The term "heterocycloalkyl", alone or in combination, denotes a monovalent saturated or partly unsaturated mono-, bi- or tricyclic ring system of 4 to 12 ring atoms, in particular 5 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Bicyclic means consisting of two cycles having one or two ring atoms in common. “Hetercycloylkyl” may comprise a carbonyl group, wherein the carbon of the carbonyl group is part of the ring system. The ring system can be attached to the remaining compound via an atom selected from C, N, S and O, in particular via a N atom (“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, piperazin-1-yl, azetidinyl, azetidin-1-yl, azetidin-3-yl, azetidin-1-yl or 5- F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 azoniaspiro[2.4]heptane. Particular examples are azetidin-3-yl, azetidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl and 5-azoniaspiro[2.4]heptane. The term “alkoxy” or “alkyloxy”, alone or in combination, signifies a group of the formula alkyl-O- in which the term "alkyl" has the previously given significance, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert.-butoxy. Particular examples of “alkoxy” are methoxy and ethoxy. The term “oxy”, alone or in combination, signifies the -O- group. The terms “halogen” or “halo”, alone or in combination, signifies fluorine, chlorine, bromine or iodine and particularly fluorine, chlorine or bromine, more particularly fluorine. The term “halo”, in combination with another group, denotes the substitution of said group with at least one halogen, particularly substituted with one to five halogens, particularly one to four halogens, i.e. one, two, three or four halogens. The term “haloalkyl”, alone or in combination, denotes an alkyl group substituted with at least one halogen, particularly substituted with one to five halogens, particularly one to three halogens, more particularly two to three halogens. Particular “haloalkyl” are fluoromethyl, fluoroethyl, fluoropropyl, fluorobutyl, difluoromethyl, difluoroethyl, trifluoromethyl and trifluoroethyl. The term “haloalkoxy”, alone or in combination, denotes an alkoxy group substituted with at least one halogen, particularly substituted with one to five halogens, particularly one to three halogens. Particular “haloalkoxy” are fluoromethoxy, fluoroethoxy and fluoropropyloxy. The terms “hydroxyl” and “hydroxy”, alone or in combination, signify the -OH group. The term “carbonyl”, alone or in combination, signifies the -C(O)- group. The term “amino”, alone or in combination, signifies the primary amino group (-NH2), the secondary amino group (-NH-), or the tertiary amino group (-N-). F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 The term “alkylamino” , alone or in combination, signifies an alkyl group linked to a -NH- group. The term “dialkylamino” denotes two alkyl groups linked to an -N- atom. The term “aminoalkyl”, alone or in combination, signifies an amino group linked to an alkyl group, wherein the alkyl group is further linked to the compound as indicated. The term “diaminoalkyl” denotes two amino groups linked to an alkyl group, wherein the alkyl group is further linked to the compound as indicated. The term “aminoalkoxy”, alone or in combination, signifies an amino group linked to an alkoxy group, wherein the alkoxy group is further linked to the compound as indicated via the - O- atom. The term “aminoalkoxyalkyl”, alone or in combination, signifies an amino group linked to an alkyl group via an alkoxy, wherein the alkyl group is further linked to the compound as indicated. The term “ammoniumalkyl”, alone or in combination, signifies an ammonium group linked to an alkyl group, wherein the alkyl group is further linked to the compound as indicated. The term “alkylcarbonyl”, is an alkyl group linked to a -C(O)- group, wherein the carbonyl group is further linked to the compound as indicated. Particular “alkylcarbonyl” are methylcarbonyl (also known as acetyl) and ethylcarbonyl. The term “carbonylalkyl”, is carbonyl group linked to an alkyl group wherein the alkyl group is further linked to the compound as indicated. Particular “alkylcarbonyl” are carbonylmethyl and carbonylethyl. The term “carbonylamino”, is carbonyl group linked to an amino group wherein the amino group is further linked to the compound as indicated. The term “sulfonyl”, alone or in combination, signifies the -SO₂- group. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 The term “pharmaceutically acceptable salts” denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts. The term “pharmaceutically acceptable acid addition salt” denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic 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 salicyclic acid. The term “pharmaceutically acceptable base addition salt” denotes those pharmaceutically acceptable salts formed with an organic or inorganic base. Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of 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, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, and polyamine resins. The term “pharmaceutically acceptable salts” includes but is not limited to the salts that have been exemplified herein. The term “pharmaceutically acceptable salts” includes those salts wherein the counter-ion has been exchanged by an otherwise suitable counter-ion, in particular by a suitable anion. The term “compound(s) of this invention” and “compound(s) of the present invention” refers to compounds of formula (I) and stereoisomers, tautomers, solvates, and salts (e.g., pharmaceutically acceptable salts) thereof. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Tautomeric forms, i.e. structural isomers which interconvert with the compound of formula (I), in particular in solution, may in some instances exist and are to be understood as being included in the invention. If one of the starting materials or compounds of formula (I) contain one or more functional groups which are not stable or are reactive under the reaction conditions of one or more reaction steps, appropriate protecting groups (as described e.g. in “Protective Groups in Organic Chemistry” by T. W. Greene and P. G. M. Wuts, 3^rd Ed., 1999, Wiley, New York) can be introduced before the critical step applying methods well known in the art. Such protecting groups can be removed at a later stage of the synthesis using standard methods described in the literature. Examples of protecting groups are tert-butoxycarbonyl (Boc), 9-fluorenylmethyl carbamate (Fmoc), 2-trimethylsilylethyl carbamate (Teoc), carbobenzyloxy (Cbz) and p- methoxybenzyloxycarbonyl (Moz). The compound of formula (I) can contain several asymmetric centers and can be present in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates. The term “asymmetric carbon atom” means a carbon atom with four different substituents. According to the Cahn-Ingold-Prelog Convention an asymmetric carbon atom can be of the “R” or “S” configuration. Furthermore, the invention includes all optical isomers, i.e. diastereoisomers, diastereomeric mixtures, racemic mixtures, all their corresponding enantiomers and/or tautomers as well as their solvates, wherever applicable, of the compound of formula (I). If desired, racemic mixtures of the compound of the invention may be separated so that the individual enantiomers are isolated. The separation can be carried out by methods well known in the art, such as the coupling of a racemic mixture of compounds to an enantiomerically pure F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 compound to form a diastereomeric mixture, followed by separation of the individual diastereomers by standard methods, such as fractional crystallization or chromatography. In the embodiments, where an optically pure enantiomer is provided, optically pure enantiomer means that the compound contains > 90 % of the desired isomer by weight, particularly > 95 % of the desired isomer by weight, or more particularly > 99 % of the desired isomer by weight, said weight percent based upon the total weight of the isomer of the compound. A chirally pure or chirally enriched compound may be prepared by chirally selective synthesis or by separation of enantiomers. The separation of enantiomers may be carried out on the final product or alternatively on a suitable intermediate. Structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Particular examples of radioisotopes are ²H, ³H, ¹³C, ¹⁴C and ¹⁸F. For example the structures wherein one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms are replaced by a ¹³C- or ¹⁴C- enriched carbon are within the scope of this invention. The invention thus relates to the following numbered aspects: 1. A compound of formula (I)

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 wherein L is absent or alkyl; X is -O- or -NH-; n is 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 heterocyloalkyl, wherein heterocycloalkyl and cycloalkyl are optionally substituted by one or two substituents independently selected from R6; with the proviso that if R1 is amino, then L is absent; R2 and R3 are independently selected from halogen; or R2 and R3 together with the carbon they are attached to form cycloalkyl; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 R4 is halogen or alkyl; R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino or piperazinyl; R6 is at each instance independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl and aminoalkylcarbonylamino; R7 is hydrogen or hydroxy; A1 is -N- or -CH-; and A₂ is -O- or -CH₂-; or a pharmaceutically acceptable salt thereof. A compound according to aspect 1, wherein L is absent. A compound according to aspect 1 or 2, wherein L is alkyl, in particular methyl. A compound according to any one of aspects 1 to 3, wherein n is 1. A compound according to any one of aspects 1 to 4, wherein X is -O-. A compound according to any one of aspects 1 to 5, wherein X is -NH-. A compound according to any one of aspects 1 to 3, wherein n is 0. A compound according to any one of aspects 1 to 7, wherein 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)aminoethylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropylamino, (hydroxy)difluoropropylamino, (hydroxy)trifluoropropylamino, F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumethoxy, cyclobutyl, azetidin-3-yl, azetidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl or 5-azoniaspiro[2.4]heptane, wherein cyclobutyl, azetidin-3-yl, azetidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl or 5- azoniaspiro[2.4]heptane are optionally substituted by one or two substituents independently selected from R6; with the proviso that if R1 is amino, then L is absent. A compound according to any one of aspects 1 to 7, wherein R1 is heterocycloalkyl optionally substituted by one or two substituents independently selected from R6. A compound according to any one of aspects 1 to 9, wherein R2 and R3 are both fluoride; or R2 and R3 together with the carbon they are attached to form cyclopropyl. A compound according to any one of aspects 1 to 10, wherein R4 is chloride or methyl, in particular chloride. A compound according to any one of aspects 1 to 10, wherein R4 is methyl. A compound according to any one of aspects 1 to 12, wherein R5 is amino or hydroxy. A compound according to any one of aspects 1 to 12, wherein R5 is amino. A compound according to any one of aspects 1 to 14, wherein R6 is at each instance independently selected from amino, methyl, hydroxy, aminomethyl, aminocarbonylmethyl and aminoethylcarbonylamino. A compound according to any one of aspects1 to 15, wherein R7 is hydrogen. A compound according to any one of aspects 1 to 16, wherein A1 is –CH-. A compound according to any one of aspects 1 to 17, A₂ is -CH₂-. A compound selected from F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; Trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidin-3-yl]propanamide; (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoro-methyl]-6-chloro- 2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide; Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide ; trans-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide; Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide; Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3- hydroxy-propanamide; Cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]urea; Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5,5-dimethyl-5- azoniaspiro[2.4]heptane-6-carboxamide; Trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5- methyl-5-azoniaspiro[2.4]heptane-6-carboxamide; Trans- (5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5- methyl-5-azoniaspiro[2.4]heptane-6-carboxamide; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]acetamide; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-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-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2-(2-aminoethoxy)acetamide; 3-amino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2- hydroxypropanamide; 2,3-diamino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4- yl)difluoromethyl)cyclohexyl)propanamide; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(diaminomethylene)urea; (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-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-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoylamino]propyl-bis(3-aminopropyl)- (carboxymethyl)ammonium; 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(3-aminopropyl)guanidine; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]-2,2,2-trifluoro-acetamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)- (carboxymethyl)ammonium; (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]morpholine-4-carboxamide; N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N-hydroxy- butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxy-propyl)amino]cyclohexyl]- difluoro-methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4- hydroxy-pyrrolidine-1-carboxamide; 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-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-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-methyl-pyrimidin-2- yl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- methyl-pyrimidin-4-yl]-difluoro-methyl]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-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)pyrrolidin-2-one; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[[4-(azetidin-1-yl)cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; tert-butyl ((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 or a pharmaceutically acceptable salt thereof. A compound according to any one of aspects 1 to 19 for use as therapeutically active substance. A pharmaceutical composition comprising a compound according to any one of aspects 1 to 19 and a therapeutically inert carrier. The use of a compound according to any one of aspects 1 to 19 for the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. The use of a compound according to any one of aspects 1 to 19 for the preparation of a medicament for the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. A compound according to any one of aspects 1 to 19 for use in the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. A method for the treatment or prophylaxis of bacterial infection, particularly an infection of a gram-negative bacteria, which method comprises administering an effective amount of a compound as defined in any one of aspects 1 to 19. The use, method or compound for use according to any one of aspects 22 to 25, wherein the gram-negative bacteria is selected from Enterobacteriaceae, Neisseria gonorrhoeae, Haemophilus influenzae, Helicobacter pylorus, Acinetobacter baumannii and Pseudomonas aeruginosa. The use, method or compound for use according to aspect 26 wherein the gram-negative bacteria is Enterobacteriaceae, wherein Enterobacteriaceae is Klebsiella pneumoniae or Escherichia coli. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 One embodiment of the invention relates to a compound according to the invention, wherein the compound is a compound of formula (II) or a pharmaceutically The synthesis of the compound of formula (I) can, for example, be accomplished according to the non-exhaustive procedures described below in general schemes 1-12 or according to methods known to those skilled in the art. In some instances, the sequence of the reaction steps can be modified and the individual steps of the different schemes can be combined in different ways as disclosed herein. All substituents are as defined in the description and in the claims, unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meaning commonly known to a person of ordinary skill in the art. Scheme 1

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 In instance benzyloxycarbonyl and tert-butoxycarbonyl group. Compound of formula (IV) can be obtained from a sulfonating reaction between nitro-phenyl sulfonyl chloride (II) and compound of formula (III). Reduction of nitro-group in compound of formula (IV) to aniline (V) can be performed by using platinium on carbon in the presence of H₂. The amidation of compound of formula (V) with a lactone (VI) in the presence of a base, such as LHMDS, affords the amide (VII). Cyclization of primary alcohol (VII) via Mitsunobu reaction gives cyclic lactam (VIII). Compound of formula (IX) can be obtained by the deprotection of compound of formula (VIII) with a reducing reagent such as palladium on carbon in the presence of H2. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Scheme 2 In A1 is -N- or -CH-; PG³ and PG⁴ are independently selected protecting groups, such as for instance tert- butoxycarbonyl and 2-(trimethylsilyl)ethoxycarbonyl group. A magnesium-halogen exchange or deprotonation of 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 addition of compound of formula (XI) affords the compound of formula (XII). Compound of formula (XIII) can be obtained from the oxidation of compound of formula (XII) in the presence of a suitable oxidant such as Dess-Martin reagent. Fluorination of compound of formula (XIII) in the presence of suitable reagent, such as DAST, can give the compound of formula (XIV). Compound of formula (XV) can be obtained by the deprotection of compound of formula (XIV) with a suitable acid such as trifluoroacetic acid. Protection of amine (XV) with PG4 by 1-[2- F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (Trimethylsilyl)ethoxycarbonyloxy]pyrrolidin-2,5-dione (Teoc-OSu) can give the compound of formula (XVI). Scheme 3 or -CH-; R is alkyl; PG³ is protecting group, such as tert-butoxycarbonyl group. Compound of formula (XVII) can be obtained by alkylation of compound of formula (XIII) and alkyl-halide such as MeI and base such as NaH can be used. Compound of formula (XVIII) can be obtained by the deprotection of compound of formula (XVII) with a suitable acid such as trifluoroacetic acid. Scheme 4

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 In A1 is -N- or -CH-. A magnesium-halogen exchange or deprotonation of 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 addition of compound of formula (XIX) affords the compound of formula (XX). Compound of formula (XXI) can be obtained from the oxidation of compound of formula (XX) in the presence of a suitable oxidant such as Dess-Martin reagent. Fluorination of compound of formula (XXI) in the presence of suitable reagent, such as DAST, can give the compound of formula (XXII). Compound of formula (XXIII) can be obtained by the reduction of compound of formula (XXII) with a suitable reducing reagent such as lithium aluminum hydride. Scheme 5 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 is alkyl; A1 is -N- or -CH-; PG⁵ is protecting groups, such as benzyloxycarbonyl group. Compound of formula (XXIV) can be obtained via Suzuki coupling between compound of formula (XXII) and alkyl-boronic acid or ester with Pd catalyst such as Pd-XPhos-G4 and base such as K3PO4. Hydrolysis of compound of formula (XXIV) in a suitable condition such as LiBr and Et₃N can give compound of formula (XXV). Curtius rearrangement of compound of formula (XXV) in the presence of DPPA and Et₃N followed by trapping of isocyanate by benzyl alcohol can give compound of formula (XXVI). Compound of formula (XXVII) can be obtained by oxidation of compound of formula (XXVI) in the presence of a suitable oxidant such as mCPBA. Scheme 6 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 is -O- or –NH-; Y is OH or leaving group such as halogen and -O-4-NO₂Ph; PG³ and PG⁶ independently are protecting groups, such as benzyloxycarbonyl and tert-butoxycarbonyl group. Compound of formula (XXIX) can be obtained by Wittig reaction of compound of formula (XIII) using suitable reagent such as methyltriphenylphosphonium bromide and base such as n-BuLi. Cyclopropanation of compound of formula (XXIX) by a suitable condition using base such as NaH in the presence of trimethylsulfoxoniumiodide can give compound of formula (XXX). Compound of formula (XXXI) can be obtained by the deprotection of compound of formula (XXX) with a suitable acid such as trifluoroacetic acid. Compound of formula (XXXIII) can be obtained by amidation of compound of formula (XXXI) with compound of formula (XXXII) using a suitable coupling condition such as HATU in the presence of base such as DIEA. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Scheme 7 CH-; R7 is H or –OH; R is alkyl or hydroxyalkyl, halogenated alkyl, halogenated alkylalcohol, cycloalkyl together is amine, cycloalkylalcohol together is amine; PG², PG³, PG⁴ and PG⁵ independently are protecting groups, such as tert-butoxycarbonyl, 2- (trimethylsilyl)ethoxycarbonyl and benzyloxycarbonyl group. The nucleophilic substitution of compound of formula (XIII, XVI, XXVII) with compound of formula (IX) in the presence of a suitable base, such as DIEA, affords compound of formula (XXXIV). Compound of formula (XXXV) can be obtained by the selective deprotection of compound of formula (XVI, XXVII) with a suitable F anion condition such as tetraethylammonium fluoroide (TEAF), or a reducing condition such as PdCl2, Et3SiH, TEA. Compound of formula (XXXVI) can be obtained by several different methods from compound of formula (XXXV) respectively; (1) 3 Step conversion to –NHOH (R7: OH) using cyanomethylation by cyanomethylbromide, mCPBA oxidation followed by NH2OH addition (2) Di-alkylation of -NH₂ by XCH₂CH(Z)CH₂X to form 4-membered ring (3) Reductive amination F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 with various aldehyde using a suitable reducing reagent such as NaBH(OAc)₃ in the presence of AcOH (4) Epoxide ring opening in the presence of base such as DIEA. Deprotection in compound of formula (XXXV) and compound of formula (XXXVI) can give compound of formula (XXXVII) and compound of formula (XXXVIII) respectively using a suitable acid such as trifluoroacetic acid. Scheme 8 independently are protecting groups, such as tert-butoxycarbonyl group. Grobal deprotection in compound of formula (XXXIV) can also give compound of formula (XXXVII) using a suitable acid such as trifluoroacetic acid. Alternatively, compound of formula (XXXVI) can be obtained by substitution of compound of formula (XVIII) with compound of formula (IX) in the presence of a suitable base, such as DIEA. Scheme 9 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 substituents (amides or carbamates); Y is OH or leaving group such as halogen and -O-4-NO2Ph; PG², and PG⁶ independently are protecting groups, such as tert-butoxycarbonyl, 2- and benzyloxycarbonyl group. In some cases, there is no protection of PG⁶. Compound of formula (XXXIX) can be obtained by two methods via coupling of compound of formula (XXXV) with compound of formula (XXXII) respectively (1) Amidation of compound of formula (XXXV) with compound of formula (XXXII) using a suitable coupling condition such as HATU in the presence of base such as DIEA (C-linked amides). (2) Y= -O-4-NO2Ph: Alcohol (HO-R1-PG⁶) is reacted with 4-nitrophenyl chloroformate to generate activated YCOO- R₁-PG⁶ (XXXII) and it is reacted with amine (XXXV) to give carbamate (XXXIX). Deprotection/global deprotection in compound of formula (XXXIX) can give compound of formula (XXXX) respectively using a suitable acid such as trifluoroacetic acid. Scheme 10 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 PG² and PG⁷ independently are protecting groups, such as tert-butoxycarbonyl, 2- and benzyloxycarbonyl group. In some cases, there is no protection of PG⁷. Compound of formula (XXXXII) can be obtained by coupling of compound of formula (XXXV) with compound of formula (XXXXI) respectively. N.N'-carbonyldiimidazole (CDI) or imidazol-1- yl(1,2,4-triazol-4-yl)methanimine are used as compound of formula (XXXXI). Substitution of amine (XXXXIII) with compound of formula (XXXXII) in the presence of base such as K₂CO₃ or no base can give compound of formula (XXXXIV) respectively. Deprotection/global deprotection in compound of formula (XXXXIV) can give compound of formula (XXXXV) using a suitable acid such as trifluoroacetic acid. Scheme 11

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 or –NH; Y is OH or leaving group such as halogen and -O-4-NO2Ph; PG² and PG⁶ independently are protecting groups, such as tert-butoxycarbonyl group. In some cases, there is no protecting group of PG⁶. Compound of formula (XXXXVI) can be obtained by substitution of compound of formula (XXIII) with compound of formula (IX) in the presence of a suitable base, such as DIEA. After conversion of primary alcohol (XXXXVI) to leaving group such as –OTf by Tf₂O in the presence of base such as DIEA, compound of formula (XXXXVII) can be obtained by substitution with NH3. Compound of formula (XXXXVIII) can be obtained by amidation of compound of formula (XXXXVII) with compound of formula (XXXII) using a suitable coupling condition such as HATU in the presence of base such as DIEA. Deprotection/global deprotection in compound of formula (XXXXVIII) can give compound of formula (XXXXIX) using a suitable acid such as trifluoroacetic acid. Scheme 12 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 In the above scheme B¹ is halide or -SO2Me; R4 is halogen or alkyl; A1 is -N- or -CH-; X is -O- or –NH; PG² and PG⁶ independently are protecting groups, such as tert-butoxycarbonyl group. In some cases, there is no protecting group of PG⁶. Compound of formula (XXXXX) can be obtained by substitution of compound of formula (XXXIII) with compound of formula (IX) in the presence of a suitable base, such as DIEA. Deprotection/global deprotection in compound of formula (XXXXX) can give compound of formula (XXXXXI) using a suitable acid such as trifluoroacetic acid. The invention thus also relates to a process for the preparation of a compound according to the invention according to the procedures described above. Pharmaceutical Compositions Another embodiment of the invention provides a pharmaceutical composition or medicament containing a compound of the invention and a therapeutically inert carrier, diluent or excipient, as well as a method of using the compounds of the invention to prepare such composition and medicament. In one example, the compound of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 buffer, at pH 5. In another embodiment, the compound of formula (I) is sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution. Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration 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 delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, epidural and intranasal, and if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents. A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., 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 formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament). The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Examples Abbreviations ACN: acetonitrile; BH3•Me2S: borane dimethyl sulfide complex; Boc: tert-butoxycarbonyl; CaCl₂: calcium chloride; Cbz: carbobenzoxyl; CDI: carbonyl diimidazole; DCE: 1,2- dichloroethane; DCE: dichloroethlyene ; DCM: dichloromethane ; DIEA: N,N- Diisopropylethylamine; DIPEA: N,N-Diisopropylethylamine; DMF: N,N-dimethylformamide; DMSO: dimethylsulfoxide; DPPA: diphenylphosphoryl azide (diphenylphosphorazidate); EA: ethyl acetate; ESI: electrospray ionization; EtOAc: ethyl acetate; Et3SiH: triethylsilane; FA: formic acid; H2O: water; HATU: Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium; HCl: hydrochloric acid; HOsu: N-hydroxylsuccinimide; HPLC: high-performance liquid chromatography; K₂CO₃: potassium carbonate; LCMS: liquid chromatography-mass spectrometry; LiHMDS: lithium bis(trimethylsilyl)amide; MeOH: methyl alcohol; m-CPBA: meta chloroperbenzoic acid; MS: mass spectrometry; MTBE: Methyl tert-butyl ether; NH₄Cl: ammonium chloride; n-Bu₃P: tributylphosphane; NaHCO₃: sodium bicarbonate; Na₂SO₄: sodium sulfate; N2: nitrogen; Na2SO3: sodium sulfite; NH2OH•HCl: hydroxylammonium chloride; NaH: sodium hydrogen; NMP: N-methyl-2-pyrrolidinone; NMR: nuclear magnetic resonance ; NH₃•H₂O: ammonium hydroxide; PdCl₂: palladium chloride; Pd/C: palladium on carbon; Pd(OH)2/C: palladium hydroxide on carbon; PE: petrol ether; Rf: retention factor; SFC: supercritical fluid chromatography; THF: tetrahydrofuran; T3P: 2,4,6-tripropyl-1,3,5,2λ⁵,4 λ⁵,6 λ⁵-trioxatriphosphinane 2,4,6-trioxide; Teoc: 2-(trimethylsilyl)ethoxycarbonyl; TEA: triethylamine; TFA: trifluoroacetic acid; TLC: thin layer chromatography. Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module. ii) ISCO combi-flash chromatography instrument. Silica gel brand and pore size: i) KP-SIL 60 Å, particle size: 40-60 µm; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400. Intermediates and final compounds were purified by preparative HPLC on reversed phase column using XBridge^TM Prep-C18 (5 µm, OBDTM 30 × 100 mm) column, SunFire^TM Prep-C18 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (5 µm, OBD^TM 30 × 100 mm) column, 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, Detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% ammonium hydroxide in water; acetonitrile and 0.1% FA in water or acetonitrile and 0.1% TFA in water). Or Gilson-281 purification System (Pump 322, Detector: UV 156, solvent system: acetonitrile and 0.05% ammonium hydroxide in water; acetonitrile and 0.225% FA in water; acetonitrile and 0.05% HCl in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water). For SFC chiral separation, intermediates were separated by chiral column (Daicel chiralpak IC, 5 µm, 30 × 250 mm), AS (10 µm, 30 × 250 mm) or AD (10 µm, 30 × 250 mm) using 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 100bar, detection UV@ 254 or 220 nm. LC/MS spectra of compounds were obtained using a LC/MS (Waters^TM Alliance 2795- Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ or Agilent Alliance 6110-Micromass ZQ), LC/MS conditions were as follows (running time 3 or 1.5 mins): Acidic condition I: A: 0.1% TFA in H₂O; B: 0.1% TFA in acetonitrile; Acidic condition II: A: 0.0375% TFA in H2O; B: 0.01875% TFA in acetonitrile; Basic condition I: A: 0.1% NH3·H2O in H2O; B: acetonitrile; Basic condition II: A: 0.025% NH₃·H₂O in H₂O; B: acetonitrile; Neutral condition: A: H₂O; B: acetonitrile. Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (MH)⁺. NMR Spectra were obtained using Bruker Avance 400 MHz. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 The microwave assisted reactions were carried out in 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 noted. Int-1-1: 2,6-dichloro-4-[1,4-dioxaspiro[4.5]decan-8-yl(difluoro)methyl]pyridine Step 1: Ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate To a solution of ethyl 4-oxocyclohexanecarboxylate (7.0 g, 41.13 mmol) and ethylene glycol (3.4 mL, 61.7 mmol) in toluene (100 mL) under N₂ was added TsOH (354 mg, 2.06 mmol). The mixture was refluxed at 120 ^oC for 18 hours. Then, the reaction mixture was cooled to room temperature and washed with aqueous NaHCO₃ (100 mL × 3). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford Int-1- 1a (8.8 g, 41.07 mmol, 99.9% yield) as light yellow oil. MS observed (ESI⁺): 215.2 [(M+H) ⁺]. Step 2: 1,4-dioxaspiro[4.5]decane-8-carboxylic acid F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-1-1b To a solution of Int-1-1a (8.8 g, 41.1 mmol) in THF (70 mL) and water (10 mL) was added lithium hydroxide monohydrate (3.45 g, 82.14 mmol). The mixture was stirred at 25 °C for 16 hours. After completion, the mixture was concentrated in vacuo to afford Int-1-1b (7.65 g, 41.08 mmol, 100% yield) as a white solid, which was used directly in the next step. MS observed (ESI⁺):187.1 [(M+H)⁺]. Step 3: N-methoxy-N-methyl-1,4-dioxaspiro[4.5]decane-8-carboxamide To a solution of Int-1-1b (7.5 g, 40.3 mmol) in DMF (100 mL) was added HATU (23.0 g, 60.42 mmol). Then O,N-dimethylhydroxylamine·HCl (5.89 g, 60.42 mmol) and TEA (16.8 mL, 120.83 mmol) were added to the reaction. The mixture was stirred at room temperature for 1 hour. After completion, the resultant mixture was diluted with EA (150 mL), washed with water (100 mL × 3). The organic layer was dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo to give crude product, which was purified by silica gel column (EA in PE =0%-10%) to afford Int-1-1c (8 g, 34.89 mmol, 86.63% yield) as colorless oil. MS observed (ESI⁺): 230.1[(M+H)⁺]. Step 4: (2,6-dichloro-4-pyridyl)-(1,4-dioxaspiro[4.5]decan-8-yl)methanone To a solution of n-BuLi (16.57 mL, 39.78 mmol) in THF (50 mL) was added a solution of 2,6-dichloropyridine (5.89 g, 39.78 mmol) in THF (20 mL) at ˗78 °C under N2. After addition, the mixture was stirred for 30 min, 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 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 was quenched with aqueous NH₄Cl solution (100 mL) and extracted with EA (150 mL × 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel column (EA in PE =0%-10%) to afford Int-1-1d (6 g, 18.98 mmol, 57.25% yield) as a light yellow solid. MS obsd. (ESI⁺):316.0 [(M+H)⁺]. Step 5: 2,6-dichloro-4-[1,4-dioxaspiro[4.5]decan-8-yl(difluoro)methyl]pyridine To a solution of Int-1-1d (4.0 g, 12.65 mmol) in DCM (20 mL) was added BAST (20 mL). The mixture was stirred at 25 °C for 3 hours. After completion, the mixture was quenched by pouring to cooled water (50 mL) slowly. The mixture was extracted with DCM (50 mL × 3). The combined organic layer was concentrated in vacuo and the crude product was purified by silica gel column (EA in PE=0%-10%) to afford Int-1-1 (3.2 g, 9.46 mmol, 74.8% yield) as a white solid. MS obsd. (ESI⁺): 338.0 [(M+H)⁺]. Int-2-1: 4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanone Compound Int-1-1 (500 mg, 1.48 mmol) was dissolved in MeOH (12 mL). Into the suspension was added with 6 M HCl (250 µL, 1.48 mmol). The reaction was stirred at 90 ^oC overnight. After completion, the reaction was cooled to room temperature and diluted with EA (30 mL) and water (30 mL). The aqueous layer was extracted with EA (20 mL × 2). The combined organic layer was washed with water (30 mL), brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude residue, which was purified F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 by silica gel column (EA in PE=0%-50%) to afford Int-2-1 (300 mg, 68.99%) as a light yellow solid. MS observed (ESI⁺) [(M+H)⁺]: 294.232. Int-3-1: 4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanamine Into the solution of Int-2 (1.74 g, 5.92 mmol) in MeOH (30 mL) was added ammonium acetate (4.56 g, 59.16 mmol) and sodium triacetoxyborohydride (2.51 g, 11.83 mmol). 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 × 3). The combined organic layer was washed with brine (50 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give the crude residue, which was purified by silica gel column (MeOH in DCM =0%-30%) to afford Int-3-1 (1.6 g, 5.42 mmol, 91.63% yield) as colorless oil. MS observed (ESI⁺):295.0[(M+H) ⁺]. Int-4-1: cis-4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanamine Int-4-1 was prepared in analogy to the preparation of compound Int-3-1 by using cis-4- [(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanone instead of Int-2-1. Int-4-1 (200 mg) was obtained as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 296.6. Int-1: tert-butyl N-[(3R)-5-oxo-1-(4-piperazin-1-ylsulfonylphenyl)pyrrolidin-3-yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 1: benzyl 4-(4-nitrophenyl)sulfonylpiperazine-1-carboxylate 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) was added 4-nitrobenzenesulfonyl chloride (50.0 g, 225.6 mmol) at 0 °C under ice-water bath. After it was stirred at 0 °C for 2 hours, the mixture was added H2O (500 mL) and it was extracted with DCM (500 mL × 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 give Int-1a (87.0 g, 214.6 mmol) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 428.1. Step 2: benzyl 4-(4-aminophenyl)sulfonylpiperazine-1-carboxylate To the solids of compound (4.0 g) were added isopropanol (400 mL) and THF (400 mL) at 25 °C under N₂. The reaction was purged with H₂ for 3 times and it was stirred at 45 °C for 72 hours under H₂ (45 Psi). After being cooled to room temperature, degassed under vacuum and purged with N2 several times, the reaction mixture was diluted with THF (1.0 L) and filtered through a celite pad, washed with THF (1.0 L). The filtrate was concentrated in vacuo to give Int-1b (81.0 g, 215.8 mmol) as a white solid. MS obsderved (ESI+) [(M+H)+]: 376.1. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 3: benzyl 4-[4-[[(3R)-3-(tert-butoxycarbonylamino)-4-hydroxy- butanoyl]amino]phenyl]sulfonylpiperazine-1-carboxylate To a solution of compound Int-1b (50.0 g, 133.2 mmol) in THF (666 mL) was added dropwise solution of LiHMDS in THF (1.0 M, 333 mL, 333 mmol) at 0 °C and it 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 it was stirred at 0 °C for 1 hour. The reaction mixture was quenched by the addition of water (500 ml) and extracted with EtOAc (500 mL × 3). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to remove solvents. The residue was purified by silica gel column (PE/EtOAc = 5/1 to 1/3; TLC: PE/EtOAc = 1/1, R_f = 0.3) to give Int-1c (55.0 g, 95.4 mmol, 71.6% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 577.3. Step 4: benzyl 4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazine-1-carboxylate 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) was added N,N,N,N-Tetramethylazodicarboxamide (CAS: 10465-78-8, 22.4 g, 130.1 mmol) in portions at 0 °C under N2. Then the mixture was warmed to 25 °C and stirred for 2 hours. After addition of aqueous HCl (1 M, 200 mL), the mixture was extracted with EtOAc (500 mL × 3). The combined organic layer was washed with saturated brine, dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to remove solvents. The residue was triturated F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 with MTBE (500 mL) to give Int-1d (45.0 g, 80.6 mmol, 92.9% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 559.2. Step 5: tert-butyl N-[(3R)-5-oxo-1-(4-piperazin-1-ylsulfonylphenyl)pyrrolidin-3- yl]carbamate To the solution of compound Int-1d (45.0 g, 80.6 mmol) in isopropanol (250 mL) and THF (250 mL) was added wet Pd/C (5.0 g, containing 50% water) at 25 °C under N2. The reaction was degassed under vacuum and purged with H2 for three times. Then the suspension was stirred at 45 °C for 12 hours under H₂ (45 Psi). After the reaction mixture was cooled to room temperature and purged by N2, the mixture was diluted with THF (1.0 L), filtered through a celite pad, washed with solution of THF (2.0 L) and MeOH (2.0 L). The filtrate was concentrated in vacuo to give crude Int-1 (38.0 g, 89.5 mmol, 94.5% yield) as a brown solid. The crude was used directly in next step without further purification. MS obsd. (ESI+) ([M+H]⁺): 425.2. Int-2: 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 Step 1: Tert-butyl N-(4-formylcyclohexyl)carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-2a To a solution of oxalyl chloride (41.5 g, 327.1 mmol) in DCM (200 mL) was added a solution of DMSO (37.5 g, 479.7 mmol) in DCM (100 mL) dropwise at -78 °C under N2, and the mixture was stirred at -78 °C for 1 hour under N2. Then the 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 stirred for 1 hour. After addition of DIEA (112.9 mL, 872.1 mmol) to the mixture at - 78 °C, the reaction was warmed to 20 °C during 1 hour and continued to be stirred at 20 °C for 1 hour. The reaction mixture was quenched with water (300 mL) at room temperature and extracted with DCM (300 mL × 3). The combined organic layer was washed with aqueous HCl (1.0 M, 100 mL × 3) and saturated brine (200 mL × 3), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give crude Int-2a (48.0 g, 211.2 mmol, 96.9% yield) as light yellow solid. The crude was used directly in next step without further purification. Step 2: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-hydroxy-methyl]cyclohexyl]carbamate To a mixture of 2,6-dichloro-4- mmol) (recrystallized by toluene in 110 °C) in THF (200 mL) was added isopropylmagnesium chloride-lithium chloride complex (268.0 mL, 348.4 mmol) dropwise under N2 at -70 ~ -40 °C. The mixture was stirred at 20 °C for 2 hours. After the solution of Int-2a (48.0 g, 211.2 mmol) in THF (200 mL) was added to the mixture at -70 ~ -40 °C, the reaction mixture was allowed to warmed to 0 °C and stirred at 0 °C for F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 0.5 hour. The reaction mixture was poured into saturated aqueous NH₄Cl (300 mL) and extracted with EtOAc (300 mL × 4). The combined organic layer was washed with saturated brine (300 mL × 3), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The resulting crude was triturated with EtOAc (150 mL) twice to give compound Int-2b (22.0 g, 58.6 mmol, 27.8% yield) as a white solid. MS obsd. (ESI+) ([{³⁵Cl}M-C4H8+H]⁺): 319.0. Step 3: Tert-butyl N-[4-(2,6-dichloropyridine-4-carbonyl)cyclohexyl]carbamate To a solution of compound Int-2b (22.0 g, 58.6 mmol) in THF (400 mL) was added DMP (49.7 g, 117.2 mmol) in portions at 0 °C. After addition of DMP, the mixture was warmed to 20 °C and stirred for 4 hours. The reaction mixture was poured into saturated aqueous K₂CO₃ (600 mL) and the mixture was stirred at 20 °C for 12 hours. The mixture was extracted with EtOAc (400 mL × 3). The combined organic layer was washed with saturated brine (150 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give Int-2c (17.0 g, 45.5 mmol, 77.7% yield) as a light yellow solid. MS observed (ESI+) ([{³⁵Cl}M-C₄H₈+H]⁺): 316.9. Step 4: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]carbamate To a solution of compound Int-2c (60 mL) was added DAST (34.0 mL) at 0 °C. After the reaction mixture was stirred at 20 °C for 16 hours, it was poured into ice F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 water (400 mL) slowly and the mixture was extracted with DCM (200 mL × 3). The combined organic layer was washed saturated aqueous NaHCO3 (200 mL), saturated brine (100 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by silica gel column (PE/EtOAc = 20/1 to 5/1; TLC: PE/EtOAc = 2/1, R_f = 0.5) to give Int-2d (11.6 g, 29.4 mmol, 64.4% yield) as a light yellow solid. MS observed (ESI+) ([{³⁵Cl}M- C4H9+H]⁺): 339.1. Step 5: 4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanamine To a solution of compound Int-2d (5.0 g, 12.7 mmol) in DCM (10 mL) was added TFA (10.0 mL) at 25 °C. After stirred at 25 °C for 1 hour, the reaction mixture was concentrated in vacuo to give crude Int-2e (3.5 g, 11.9 mmol, 93.7% yield) as dark green oil. The crude was used directly in next step without purification. MS obsd. (ESI+) ([{³⁵Cl}M+H]⁺): 294.8. Step 6: 2-(trimethyl-λ4-sulfanyl)ethyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoro- methyl]cyclohexyl]carbamate To a solution of compound (6.2 mL, 35.6 mmol) in DCM (50 mL) was added Teoc-OSu (3.7 g, 14.2 mmol) at 0 °C. After 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 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 saturated brine (100 ml). The combined organic layer was 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 give Int-2f (3.6 g, 8.1 mmol, 85.9% yield) as yellow oil. MS obsd. (ESI+) ([{³⁵Cl}M-C₂H₄+H]⁺): 411.2. Step 7: 2-trimethylsilylethyl N-[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert- butoxycarbonylamino)-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4-pyridyl]- difluoro-methyl]cyclohexyl]carbamate 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) in one portion. After being stirred 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 layer was dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give Int-2g (1.5 g, 1.8 mmol, 97.9% yield) as yellow oil. The crude product was used in next step directly without further purification. MS observed (ESI+) ([{³⁵Cl}M+H]⁺): 827.5, ([{³⁵Cl}M-C2H4+H]⁺): 799.5, ([{³⁵Cl}M-Boc-C₂H₄+H]⁺): 699.5. Step 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 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of compound Int-2g (1.5 g, 1.8 mmol) in ACN (20 mL) was added tetraethylammonium fluoroide (6.7 g, 36.3 mmol) in one portion. After being stirred at 50 °C for 2 hours, the mixture was cooled to room temperature and adjusted to pH 8 ~ 9 with aqueous NaHCO₃. The solution was extracted with EtOAc (100 mL × 2) and washed with saturated brine (50 mL × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give crude Int-2 (550.0 mg, 0.8 mmol, 44.4% yield) as a yellow solid. The crude was used directly in next step without further purification. MS obsd. (ESI+) ([{³⁵Cl}M+H]⁺): 683.1 Int-3: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]carbamate Int-3 was prepared in analogy to 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 in step 1. Int-3 (200 mg) was obtained as a white solid. MS obsd. (ESI+) ([{³⁵Cl}M-C4H9+H]⁺): 339.1, ([{³⁵Cl}M-Boc+H]⁺): 294.8. Int-4: Tert-butyl N-[rac-(3R)-1-[4-[4-[4-[[4-(aminomethyl)cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Step 1: F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of 4-(methoxycarbonyl)cyclohexane-1-carboxylic acid (50.0 g, 268.5 mmol) in THF (150 mL) was added BH₃•Me₂S (10 M, 54 mL, 540 mmol) dropwise at 0 ~ 10 °C for 1 hour under N2. The mixture was stirred at 25 °C for 12 hours. After the reaction mixture was cooled to 0°C, MeOH (100 mL) was added into the reaction mixture dropwise at 0 °C to quench the reaction. The mixture was concentrated in vacuo to dryness at 40 °C. The residue was purified by silica gel column (PE/EtOAc = 100/1 to 1/1; TLC: PE/EtOAc = 2/1, R_f = 0.4) to give Int-4a (28.0 g, 162.6 mmol, 60.6% yield) as colorless oil. The Int-4a was used in next step directly without any analysis. It was determind by comparing the R_f of commercially available standard sample on TLC.¹H NMR (400 MHz, DMSO-d₆) δ ppm 3.58 (s, 3 H), 3.19 - 3.22 (d, J = 4 Hz, 2H), 2.22 - 2.23 (m, 1 H), 1.77 - 1.92 (m, 2 H), 1.73 - 1.75 (m, 2 H), 1.27 -1.32 (m, 3 H), 0.91- 0.96 (td, J = 9.6, 3.6 Hz, 2 H). MS observed (ESI+) [(M+H)+]: 173.2. Step 2: Methyl 4-formylcyclohexanecarboxylate To a solution of oxalyl chloride (31.0 g, 243.9 mmol) in DCM (100 mL) was added DMSO (25.4 g, 325.2 mmol) at -60 °C and the mixture was stirred at -60 °C for 0.5 hour under N₂. After the solution of compound Int-4a (28.0 g, 162.6 mmol) in DCM (100 mL) was added at -60 °C, the reaction mixture was stirred at -60 °C for 1 hour. Then DIEA (84.1 g, 650.3 mmol) was added at - 60 °C and the mixture was continued to be stirred at -60 ~ -20 °C for 1 hour. Next, the mixture was warmed to 0 °C and stirred for another 1 hour under N2. The mixture was diluted with EtOAc (600 mL) and washed with aqueous CaCl2 (100 mL × 3) and saturated brine (100 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by silica gel F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 column (PE/EtOAc = 30/1 to 1/1; TLC: PE/EtOAc = 3/1, R_f = 0.5) to give Int-4b (24.0 g, 141.0 mmol, 86.7% yield) as light yellow oil. The int-4b was used in next step directly without characterization analysis. It was determind by comparing the R_f of commercially available standard sample on TLC.¹H NMR (400 MHz, DMSO-d6) δ ppm 9.56 - 9.58 (s, 1 H), 2.24 - 2.29 (m, 2 H), 1.93 - 2.00 (m, 4 H), 1.26 - 1.34 (m, 4 H) Step 3: 4-[(2,6-dichloro-4-pyridyl)-hydroxy-methyl]cyclohexanecarboxylate To a solution of 2,6-dichloro-4-iodopyridine (42.5 g, 155.1 mmol) in THF (50 ml) was added isopropylmagnesium chloride lithium chloride complex in THF (130.2 mL, 169.2 mmol) dropwise at -40 °C under N₂. The mixture was stirred at 25 °C for 1 hour. Then the mixture was added into the solution of compound Int-4b (24.0 g, 141.0 mmol) in THF (100 mL) dropwise at -40 °C. The reaction mixture was continued to be stirred at 25 °C for 3 hours. The mixture was poured into aqueous NH₄Cl (200 mL) and extracted with EtOAc (300 mL × 3). The combined organic layer was washed with saturated brine (100 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by silica gel column (PE/EtOAc = 100/1 to 1/2; TLC: PE/EtOAc = 1/1, Rf = 0.4) to give compound Int-4c (33.0 g, 103.7 mmol, 73.6% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 317.8. Step 4: Methyl 4-(2,6-dichloropyridine-4-carbonyl)cyclohexanecarboxylate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of compound Int-4c (33.0 g, 103.7 mmol) in THF (150 mL) was added Dess-Martin periodinane (48.4 g, 114.1 mmol) slowly at 0 °C under N2. Then the mixture was warmed to 25 °C and continued to be stirred at this temperature for 3 hours under N₂. The mixture was diluted with EtOAc (500 mL) and poured into aqueous K₂CO₃ (200 mL). The mixture was stirred at room temperature for 0.5 hour and filtered. The organic layer was separated and washed with aqueous K2CO3 (100 mL × 2) and saturated brine (100 mL × 2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give crude compound Int-4d (30.0 g, 94.9 mmol, 91.5% yield) as a white solid. The crude product was used in next step directly without further purification. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 315.8. Step 5: Methyl 4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexanecarboxylate To a solution of compound Int-4d (29.5 g, 93.3 mmol) in DCM (20 mL) was added diethylaminosulfur trifluoride (135.4 g, 839.7 mmol) and it was stirred at 45 °C for 12 hours. After being cooled to 25 °C, the mixture was poured into ice water at 0 °C and then extracted with EtOAc (200 mL × 3). The combined organic layer was washed with aqueous K2CO3 (100 mL × 2) and saturated brine (100 mL × 2), dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated in vacuo. The residue was recrystallized with MeOH (150 mL) to give compound Int-4e (25.6 g, 75.7 mmol, 81.1% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 337.8. Step 6: [4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]methanol F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-4f To a solution of compound Int-4e (250.0 mg, 0.7 mmol) in THF (5 mL) was added LiAlH4 (42.1 mg, 1.1 mmol) at -10 °C and it was stirred at -10 °C for 2 hours. After the mixture was warmed to 0 ℃, the reaction mixture was diluted with THF (100 mL) and quenched with H₂O (0.1 mL), followed by 15% aqueous NaOH (0.2 mL) and H2O (0.1 mL) successively. The suspension was added MgSO4 (10 g) and stirred at 20 °C for 0.5 hour. The mixture was filtered through a celite pad, and the filtrate was concentrated in vacuo. The residue was purified by prep-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 give Int-4f (210.0 mg, 0.7 mmol, 91.6% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 310.1 Step 7: Tert-butyl N-[rac-(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4- (hydroxymethyl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo- pyrrolidin-3-yl]carbamate To a solution of compound Int-4f (210.0 mg, 0.7 mmol) in DMSO (1 mL) was added Int-1 (288.1 mg, 0.7 mmol) at 20 °C and it was stirred at 120 °C for 16 hours. After being cooled to room temperature, the resulting mixture was transferred to a separatory funnel and the aqueous layer was extracted with EtOAc (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by prep-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 give Int-4g (400.0 mg, 0.6 mmol, 77.4% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]:: 698.2. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 7: Tert-butyl N-[rac-(3R)-1-[4-[4-[4-[[4-(aminomethyl)cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate To a solution of compound Int-4g (380.0 mg, 0.5 mmol) in DCM (5 mL) was added trifluoromethanesulfonic anhydride (0.2 mL, 1.1 mmol) at -20 °C and it was stirred at -20 °C for 0.5 hour. Then the mixture was poured into solution of ammonia in THF (20 mL) and stirred for 1 hour at 45 °C. After the mixture was concentrated to 10 mL, it was diluted by slow addition of saturated aqueous ammonium chloride (20 mL). The resulting mixture was transferred to a separatory funnel and the aqueous layer mixture was extracted with EtOAc (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4, filtered. The filtrate was concentrated under reduced pressure to afford the crude product, which was purified by prep-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 give Int-4 (120.0 mg, 0.2 mmol, 31.6% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 697.2. Int-5: tert-butyl ((R)-1-(4-((4-(4-(((1r,4r)-4-aminocyclohexyl)difluoromethyl)-6- methylpyrimidin-2-yl)piperazin-1-yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate Step 1: Chlorolithium;chloro-(2,2,6,6-tetramethyl-1-piperidyl)magnesium F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of isopropyl magnesium chloride-lithium chloride complex (50.0 mL, 65.0 mmol) was added 2,2,6,6-tetramethylpiperidine (12.1 mL, 71.5 mmol) dropwise at -40 °C. After the mixture was stirred at 25 °C for 24 hours under N2, the solution (60 mL, ~ 1.1 M) of Int-5a in THF was obtained. The solution was used in next step directly without further operation. Step 2: Methyl 4-[(6-chloro-2-methylsulfanyl-pyrimidin-4-yl)-hydroxy- methyl]cyclohexanecarboxylate To a solution of 4-chloro-2-(methylthio)pyrimidine (3.0 g, 18.7 mmol) in THF (8 mL) was added Int-5a (18.7 mL, 20.6 mmol) at -40 °C and it was stirred at 0 °C for 40 min. After the mixture was added into the solution of methyl 4-formylcyclohexanecarboxylate (3.5 g, 20.6 mmol) in THF (12 mL) at -40 °C, it was warmed to 25 °C and stirred for 1 hour. The mixture was poured into water (50 mL) and extracted with EtOAc (80 mL × 3). The organic layer was washed with saturated brine (50 mL × 2), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel column (PE/EtOAc = 100/1 to 1/1; TLC: PE/EtOAc = 1/1, Rf = 0.4) to give Int-5b (1.8 g, 5.4 mmol, 29.1% yield) as light yellow oil. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 331.3 Step 3: Methyl 4-(6-chloro-2-methylsulfanyl-pyrimidine-4-carbonyl)cyclohexanecarboxylate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of compound Int-5b (1.7 g, 5.1 mmol) in THF (30 mL) was added Dess-Martin periodinane (2.6 g, 6.2 mmol) at 0 °C and it was stirred at 25 °C for 2 hours. The mixture was quenched with ice water (30 mL) and adjusted to pH = 9 with aqueous NaHCO3. The mixture was extracted with EtOAc (100 mL × 2) and washed with saturated brine (50 mL × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column (PE/EtOAc = 1/0 to 1/1; TLC: PE/EtOAc = 1/1, Rf = 0.5) to give Int-5c (1.6 g, 4.9 mmol, 94.7% yield) as a yellow solid. (ESI⁺) [({³⁵Cl}M+H)⁺]: 329.3. Step 4: Methyl 4-[(6-chloro-2-methylsulfanyl-pyrimidin-4-yl)-difluoro- methyl]cyclohexanecarboxylate To a solution of Int-5c (1.6 g, 4.9 mmol) in DCM (10 mL) was added diethylaminosulfur trifluoride (10.0 mL, 4.9 mmol) at 0 °C and it was warmed to 25 °C and stirred for 2 hours. The mixture was poured into ice water carefully and adjusted to pH = 9 with aqueous NaHCO₃. The mixture was extracted with EtOAc (200 mL × 2) and washed with saturated brine (100 mL × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column (PE/EtOAc = 1/0 to 5/1; TLC: PE/EtOAc = 5/1, R_f = 0.55) to give Int-5d (1.5 g, 4.3 mmol, 87.9% yield) as a yellow solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 351.3. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 5: Methyl 4-[difluoro-(6-methyl-2-methylsulfanyl-pyrimidin-4- yl)methyl]cyclohexanecarboxylate 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 potassiumphosphate (1.6 g, 7.7 mmol) in 1,4-dioxane (5 mL) and water (2 mL) was added trimethylboroxine (9.7 mL, 30.8 mmol) at 25 °C. The reaction mixture was degassed under vacuum and purged with N² several times and it was stirred at 80 °C for 12 hours under N2. After being cooled to room temperature, the mixture was poured into H₂O (20 ml), extracted with EtOAc (50 mL × 2) and washed with brine (50 ml × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column (PE/EtOAc = 30/1 to 10/1; TLC: PE/EtOAc = 5/1, Rf = 0.65) to give the Int-5e (420.0 mg, 1.3 mmol, 34.7% yield) as yellow oil. MS observed (ESI⁺) [(M+H)⁺]: 331.3. Step 6: (1r,4r)-4-(difluoro(6-methyl-2-(methylthio)pyrimidin-4-yl)methyl)cyclohexane-1- carboxylic acid: To a mixture of Int-5e (2.3 g, and water (2 mL) were added LiBr (6.0 g, 70.0 mmol) and TEA (2.1 g, 21.0 mmol) at 25 °C. After the reaction was stirred at 50 °C for 2 hours, the mixture was quenched by slow addition of HCl solution (1.0 M, 20 mL). The F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 resulting mixture was transferred to a separatory funnel and the aqueous layer was extracted with EtOAc (100 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford the crude product Int-5f (2.0 g, 6.3 mmol, 90.3% yield) as a white solid, which was used in next step without further purification. MS observed (ESI⁺) [(M+H)⁺]: 317.1. Step 7: 4-(difluoro((1r,4r)-4-isocyanatocyclohexyl)methyl)-6-methyl-2- (methylthio)pyrimidine To a solution of Int-5f (2.0 g, 6.3 mmol) in THF (10 mL) were added DPPA (3.7 g, 13.3 mmol) and TEA (2.2 g, 22.1 mmol) at 25 °C. After the reaction was stirred at 25 °C for 2 hours, the mixture was quenched by slow addition of saturated aqueous ammonium chloride (20 mL). The resulting mixture was transferred to a separatory funnel, and the aqueous layer was extracted with EtOAc (100 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column (PE/EtOAc = 1/0 to 2/1; TLC: PE/EtOAc = 2/1, Rf = 0.5) to give the Int-5g (1.0 g, 3.2 mmol, 50.5% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 314.0 Step 8: Benzyl ((1r,4r)-4-(difluoro(6-methyl-2-(methylthio)pyrimidin-4- yl)methyl)cyclohexyl)carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-5h Benzyl alcohol (20.0 mL) was added to Int-5g (1.0 g, 3.2 mmol) at 25 °C and it was heated to 50 °C stirred for 2 hours. After cooling to 25 °C, the resulting mixture poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with EtOAc (80 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column (PE/EtOAc = 1/0 to 2/1; TLC: PE/EtOAc = 1/1, R_f = 0.3) to give the Int-5h (1.1 g, 2.6 mmol, 81.8% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 422.1. Step 9: Tert-butyl ((1r,4r)-4-(difluoro(6-methyl-2-(methylsulfonyl)pyrimidin-4- yl)methyl)cyclohexyl)carbamate To a solution of Int-5h (1.1 were added sodium tungstate dihydrate (172.2 mg, 0.5 mmol) and hydrogen peroxide (4.0 mL, 30% wt.) at 0 °C in one portion under N2. After the reaction was stirred at 25 °C for 16 hours, it was quenched with aqueous NaHCO₃ (100 mL), aqueous Na₂SO₃ (100 mL) and then extracted with EtOAc (100 mL × 3). The combined organic layer was washed with saturated brine (100 mL), dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated in vacuo to afford the crude product, which was purified by silica gel column (PE/EtOAc = 1/0 to 1/1; TLC: PE/EtOAc = 1/1, R_f = 0.5) to give the Int-5i (1.0 g, 2.2 mmol, 84.7% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 454.1. Step 10: Tert-butyl ((R)-1-(4-((4-(4-(((1r,4r)-4- (((benzyloxy)carbonyl)amino)cyclohexyl)difluoromethyl)-6-methylpyrimidin-2- yl)piperazin-1-yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of Int-5i (1.0 g, 2.2 mmol) in DMSO (4 mL) were added DIEA (0.9 g, 6.6 mmol) and Int-1 (1.0 g, 2.4 mmol). After the reaction was stirred at 100 °C for 16 hours, it was 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 EtOAc (80 mL × 3). The combined organic layer was washed with brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column (PE/EtOAc = 1/0 to 0/1; TLC: PE/EtOAc = 1/1, Rf = 0.4) to give the Int-5j (1.2 g, 1.5 mmol, 67.9% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 798.6. Step 11: Tert-butyl ((R)-1-(4-((4-(4-(((1r,4r)-4-aminocyclohexyl)difluoromethyl)-6- methylpyrimidin-2-yl)piperazin-1-yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate To a solution of Int-5j (1.2 g, 1.5 mmol) in DCM (20 mL) were added Et3SiH (1.9 mL, 12.0 mmol), PdCl2 (132.4 mg, 0.8 mmol) and TEA (0.6 mL, 4.5 mmol). The mixture was degassed under vacuum and purged with N2 for 3 times. After the reaction was stirred at 25 °C for 2 hours under N2, the resulting mixture was poured into water (50 mL) and transferred to a separatory funnel. The aqueous layer was extracted with EtOAc (100 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (EtOAc/MeOH = 10:0 to 5:1; TLC: EtOAc/MeOH = 5/1, R_f = 0.5) to give the Int-5 (870.0 mg, 1.3 mmol, 87.2% yield) as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 664.3. Int-6: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4- (hydroxyamino)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo- pyrrolidin-3-yl]carbamate Step 1: Benzyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoro- methyl]cyclohexyl]amino]propyl]carbamate To a solution of Int-2 (0.3 g, 0.4 mmol) in DMF (2 mL) were added bromoacetonitrile (105.3 mg, 0.9 mmol) and K₂CO₃ (181.8 mg, 1.3 mmol). After the reaction was stirred at 20 °C for 16 hours, it was diluted with EtOAc (50 mL) and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-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 give Int-6a (150.0 mg, 0.2 mmol, 47.3% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 722.3. Step 2: N-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-1-cyano- methanimine oxide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of compound Int-6a (200.0 mg, 0.3 mmol) in DCM (3 mL) was added m-CPBA (105.1 mg, 0.6 mmol) at 0 °C. After being stirred at 20 °C for 2 hours, the reaction mixture was quenched with saturated aqueous Na2SO3 solution (10 mL) and the reaction mixture was extracted with DCM (70 mL × 3). The combined organic layer was washed with NaHCO3 (50 mL), saturated brine (50 mL × 3) successively and dried over anhydrous Na₂SO₄ and concentrated in vacuo. The residue was purified by prep-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 give Int-6b (100.0 mg, 0.1 mmol, 49.1% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 736.3. Step 3: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4- (hydroxyamino)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo- pyrrolidin-3-yl]carbamate To a solution of compound Int-6b (60.0 mg, 0.1 mmol) in methanol (1.0 mL) and DMF (1 mL) was added NH2OH•HCl (34.0 mg, 0.5 mmol) at 20 °C and it was stirred at 60 °C for 2 hours. The solvent was concentrated in vacuo and the residue was purified by prep-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 give Int-6 (50.0 mg, 0.1 mmol, 87.8% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 699.3. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-7: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(methylamino)cyclohexyl]methyl]- 2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Step 1: Tert-butyl N-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]-N-methyl- carbamate To a solution of Int-2d (0.8 g, 2.0 mmol) and iodomethane (316.0 mg, 2.2 mmol) in DMF (2 mL) was added NaH (89.1 mg, 2.2 mmol) at 0 °C and it was stirred at 0 °C for 1 hour. After the reaction quenched by addition of ice water (10 mL), the mixture was extracted with EtOAc (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated in vacuo to afford crude product, which was purified by prep-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 give Int-7a (541.0 mg, 1.3 mmol, 65.3% yield) as yellow oil. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 353.2. Step 2: 4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]-N-methyl-cyclohexanamine F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of Int-7a (500.0 mg, 1.2 mmol) in DCM (1 mL) was added TFA (1.0 mL, 13.0 mmol) at 25 °C and it was stirred at this temperature for 1 hour. The mixture was concentrated in vacuo and the residue was purified by prep-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 give int.7b (320.0 mg, 1.0 mmol, 84.7% yield) as yellow oil. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 309.0. Step 3: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4- (methylamino)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo- pyrrolidin-3-yl]carbamate 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) was added DIEA (190.0 mg, 1.5 mmol) and it was stirred at 100 °C for 16 hours. After being cooled to 25 °C, the mixture was concentrated in vacuo to give a residue, which was purified by prep- 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 give Int-7 (240.0 mg, 0.3 mmol, 71.0% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 697.4. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 001: Cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one (001A) and Trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one (001B) Step 1: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[1,4-dioxaspiro[4.5]decan-8- yl(difluoro)methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3- yl]carbamate Compound Int-1-1 (200 mg, 591 µmol) and tert-butyl (R)-(5-oxo-1-(4-(piperazin-1- ylsulfonyl)phenyl)pyrrolidin-3-yl)carbamate (213 mg, 502 µmol) were dissolved in the DMF (6 mL), followed by adding DIEA (263 µl, 1.51 mmol). The reaction was stirred at 120 °C overnight. After the reaction was 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 × 2). The combined organic layer was washed with water (30 mL), brine (30 mL), dried over anhydrous F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Na2SO4, filtered and concentrated in vacuo to give the crude residue, which was purified by silica gel column (EA in PE=0%-70%) to afford 001b (0.29 g, 399 µmol, 79.6 % yield). MS observed (ESI⁺): 726.201 [(M+H)⁺]. Step 7: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-(4-oxocyclohexyl)methyl]-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Compound 001a (227 mg, 313 µmol) was dissolved in the 36% HCl (5 mL, 25 mmol) and MeOH (5 mL). The mixture was stirred at 50 °C for 2 hours. LCMS showed mainly product formed while de-Boc product was also observed. Then, the mixture was adjusted with aqueous NaHCO3 solution to pH 8 and extracted with DCM (30 mL × 3). The combined organic layer was concentrated in vacuo to give crude product (180 mg, 201 µmol, 64.3 % yield) as a light yellow solid. MS observed (ESI⁺): 582.375 [(M+H)⁺]. The resultant compound was dissolved in the DCM (2.5 mL). Into the stirring solution was added Boc2O (93.7 mg, 430 µmol) and Et3N (71.8 µl, 515 µmol). The reaction was stirred at room temperature overnight and LCMS showed the starting material went completed. After completion, the reaction was extracted with DCM (30 mL × 3). The combined organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 001b (100 mg, 147 µmol, 85.3 % yield) as a yellow oil. MS observed(ESI⁺): 682.152 [(M+H)⁺]. Step 8: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3- yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 001c Compound 001b (100 mg, 172 µmol) and azetidin-3-ol hydrochloride (94.1 mg, 859 µmol) were dissolved in the THF (15 mL), followed by adding one drop of titanium (IV) isopropoxide. 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 stirred at 70 ^oC for 2 hours. After completion, the mixture was poured into water (50 mL), and extracted with EA (50 mL × 3). The combined organic layer was washed with brine (50 mL × 2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give crude 001c (127 mg, 172 µmol, 100% yield), which was used directly in the next step. MS observed (ESI⁺): 739.3[(M+H)⁺]. Step 9: Cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one (001A) and Trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one (001B) Compound 001c (127 mg, 172 µmol) and TFA (23.1 mg, 203 µmol) were mixed in the DCM (5 mL). The reaction was stirred at room temperature for 1 hour. After completion, the reaction was concentrated in vacuo to give crude compound, which was purified by prep-HPLC to give a mixture of 001A and 001B. The two epimers were separated by SFC to give Example 001A F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (eluted first, 93 mg, 64% yield) and Example 001B (eluted second, 14 mg, 20% yield). The cis and trans configurations were confirmed by 2D NMR. Example 001A: MS obsd. (ESI⁺) [(M+H)⁺]: 639.3.¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.88 - 7.98 (m, 2 H), 7.76 - 7.85 (m, 2 H), 6.71 - 6.81 (m, 1 H), 6.64 - 6.69 (m, 1 H), 4.36 - 4.51 (m, 1 H), 4.18 - 4.36 (m, 3 H), 4.04 - 4.15 (m, 1 H), 3.74 - 3.95 (m, 3 H), 3.59 - 3.73 (m, 4 H), 3.02 - 3.15 (m, 1 H), 2.88 - 3.02 (m, 4 H), 2.54 - 2.66 (m, 1 H), 2.17 - 2.34 (m, 1 H), 1.67 - 1.82 (m, 2 H), 1.52 - 1.67 (m, 2 H), 1.32 - 1.52 (m, 4 H).¹⁹F NMR (376 MHz, DMSO-d₆) δ ppm -105.95 - - 101.82 (m, 2 F). Example 001B: MS obsd. (ESI⁺) [(M+H)⁺]: 683.2.¹H NMR (400 MHz, DMSO-d6) δ ppm 7.87 - 8.00 (m, 2 H), 7.73 - 7.83 (m, 2 H), 6.76 - 6.83 (m, 1 H), 6.69 - 6.75 (m, 1 H), 6.14 - 6.32 (m, 1 H), 4.37 - 4.50 (m, 1 H), 4.18 - 4.34 (m, 3 H), 4.00 - 4.18 (m, 1 H), 3.74 - 3.89 (m, 3 H), 3.61 - 3.72 (m, 4 H), 3.02 - 3.13 (m, 2 H), 2.90 - 3.02 (m, 4 H), 2.53 - 2.65 (m, 1 H), 2.09 - 2.30 (m, 1 H), 1.87 - 2.04 (m, 2 H), 1.59 - 1.81 (m, 2 H), 0.98 - 1.20 (m, 4 H).¹⁹F NMR (376 MHz, DMSO- d6) δ ppm -108.32 - -100.42 (m, 2 F). Example 002: 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidin- 3-yl]propanamide Step 1: Tert-butyl N-[1-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]azetidin-3- yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 002a Compound Int-2-1 (300 mg, 1.02 mmol), N-(azetidin-3-yl)carbamic acid tert-butyl ester (263.49 mg, 1.53 mmol) and sodium triacetoxyborohydride (648.53 mg, 3.06 mmol) were added into the mixture of DCM (15 mL) and MeOH (5 mL). The reaction was stirred at room temperature for 1 day. LCMS showed full conversion of the starting material. Then the mixture was poured into water (50 mL), and extracted with EA (50 mL × 3). The combined organic layer was washed with brine (50 mL × 2), dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude 002a (400 mg, 87.08%) as a yellow solid. MS obsd. (ESI⁺) [(M+H)⁺]: 450.22. Step 3: 1-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]azetidin-3-amine Compound 002a (300 mg, 0.666 mmol) and TFA (1.5 mL, 20.22 mmol) were dissolved in the DCM (15 mL). The reaction was stirred at room temperature for 2 hours. After completion, the reaction was concentrated in vacuo. The resulting residue 002b was azerotroped with toluene and used directly in the next step. MS obsd. (ESI⁺) [(M+H)⁺]:350.8. Step 4: Tert-butyl N-[2-[[1-[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]azetidin- 3-yl]amino]-1-methyl-2-oxo-ethyl]carbamate Compound 002b (130 mg, 0.280 mmol) and 2-(tert-butoxycarbonylamino)propionic acid (66.23 mg, 0.350 mmol) were dissolved in the DCM (15 mL). Into the stirring solution was added HATU (159 mg, 0.420 mmol) and DIEA (146.71 uL, 0.840 mmol). The reaction was F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 stirred at room temperature for 2 hours. After completion, the reaction was diluted with DCM (15 mL) and water (15 mL). The aqueous layer was extracted with DCM (50 mL × 3). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give crude 002c (120 mg, 82.19% yield) as a yellow solid, which was used directly in the next step. Step 5: Tert-butyl N-[2-[[1-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]azetidin-3-yl]amino]-1-methyl-2-oxo-ethyl]carbamate Compound 002c (120 mg, 0.230 mmol) was dissolved in DMF (10 mL), followed by adding N- [(3R)-5-keto-1-(4-piperazinosulfonylphenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (97.7 mg, 0.230 mmol) and DIEA (120 µL, 0.690 mmol). The mixture was stirred at 125 ^oC overnight. After full conversion of starting material, the reaction mixture was diluted with water (50 mL) and extracted with EA (50 mL × 2). The combined organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4 and concentrated to dryness to give crude 002d (150 mg, 71.67% yield) as yellow oil, which was used directly in the next step. MS observed (ESI⁺) [(M+H)⁺]: 909.6. Step 6: 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidin- 3-yl]propanamide (002) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 002 Compound 002d (100 mg, 0.110 mmol) was dissolved in the DCM (5 Ml), followed by adding TFA (2 mL, 25.96 mmol). The reaction was stirred at room temperature for 2 hours. After completion, the reaction was concentrated under vacuum and the resultant residue was purified by prep-HPLC to give Example 002 (7.3 mg, 7.26% yield) as a light yellow solid. MS obsd. (ESI⁺) [(M+H)⁺]: 709.3.¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.79 - 7.89 (m, 2 H), 7.68 - 7.78 (m, 2 H), 7.39 - 7.49 (m, 1 H), 6.50 - 6.64 (m, 1 H), 4.35 - 4.46 (m, 1 H), 4.20 - 4.32 (m, 1 H), 3.99 - 4.11 (m, 1 H), 3.86 - 3.97 (m, 1 H), 3.72 - 3.86 (m, 2 H), 3.56 - 3.63 (m, 4 H), 3.32 - 3.42 (m, 1 H), 3.01 - 3.11 (m, 1 H), 2.88 - 3.01 (m, 4 H), 2.47 - 2.62 (m, 1 H), 2.00 - 2.13 (m, 1 H), 1.86 - 1.96 (m, 1 H), 1.72 - 1.82 (m, 1 H), 1.59 - 1.72 (m, 2 H), 1.33 - 1.50 (m, 8 H), 1.13 - 1.33 (m, 3 H).¹⁹F NMR (376 MHz, METHANOL-d4) δ ppm -113.96 - -100.71 (m, 2 F). Example 003: (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoro- methyl]-6-chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one Step 1: benzyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoro- methyl]cyclohexyl]amino]propyl]carbamate Into a solution of Int-3-1 (313 mg, 1.06 mmol) in DCM (5 mL) was added benzyl (3- oxopropyl)carbamate (330 mg, 1.59 mmol) and sodium triacetoxyhydroborate (674 mg, 3.18 mmol). The reaction was stirred at 25 °C overnight. After completion, the reaction mixture was F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 poured into water, extracted with DCM (50 mL × 3). The combined organic layer was concentrated under vacuum to give crude 003a (500 mg, 96% yield) as yellow oil, which was used directly in the next step. MS obsd. (ESI⁺) [(M+H)⁺]: 486.1. Step 2: Tert-butyl N-[3-(benzyloxycarbonylamino)propyl]-N-[4-[(2,6-dichloro-4-pyridyl)- difluoro-methyl]cyclohexyl]carbamate Compound 003a (86 mg, 177 µmol) was dissolved in the DCM (10 mL). Into the stirring solution was added Boc₂O (117 mg, 124 µl, 535 µmol) and triethylamine (112 µl, 802 µmol). The mixture was stirred at room temperature overnight. After completion, the reaction mixture was poured into water, extracted with DCM (50 mL × 3). The combined organic layer was concentrated under vacuum to give crude 003b, which was used directly in the next step. MS obsd. (ESI⁺) [(M+H)⁺]: 586.1. Step 3: Tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[3- (benzyloxycarbonylamino)propylamino]cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Compound N-[(3R)-5-keto-1-(4-piperazinosulfonylphenyl)pyrrolidin-3-yl]carbamic acid tert- butyl ester (100 mg, 0.236 mmol) and 003b (150 mg, 0.256 mmol) were dissolved in the DMF (5 mL). Into the solution was added DIEA (134. uL, 0.767 mmol). The reaction was stirred at 120 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 ^oC overnight. After completion, the reaction was cooled to room temperature and diluted with EA (100 mL). The aqueous layer was extracted with EA (150 mL × 3). The combined organic layer was washed with water (100 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give crude 003c (200 mg, 80.24%) as light brown oil, which was used directly in the next step. Step 4: (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one Compound 003c (100 mg, 0.103 mmol) were mixed with TFA (2 mL). The mixture was heated at 90 ^oC for 2 hours. After completion, the reaction mixture was concentrated under reduced pressure to give crude residue, which was purified by prep-HPLC to give Example 003 (2.5 mg, 3.43% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 640.4.¹H NMR (400 MHz, METHANOL-d4) δ ppm 7.89 - 7.99 (m, 2 H), 7.79 - 7.89 (m, 2 H), 7.48 - 7.62 (m, 1 H), 6.60 - 6.75 (m, 1 H), 5.29 - 5.39 (m, 1 H), 4.34 - 4.45 (m, 1 H), 4.12 - 4.25 (m, 1 H), 3.88 - 3.99 (m, 1 H), 3.62 - 3.75 (m, 4 H), 3.18 - 3.24 (m, 1 H), 2.99 - 3.18 (m, 8 H), 2.14 - 2.30 (m, 3 H), 1.99 - 2.14 (m, 3 H), 1.76 - 1.99 (m, 3 H), 1.54 - 1.76 (m, 2 H), 1.37 - 1.52 (m, 2 H).¹⁹F NMR (376 MHz, METHANOL-d₄) δ ppm -112.27 - -105.22 (m, 2 F). Example 004: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-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- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide (004B) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 1: Tert-butyl N-[3-[[4-[(2,6-dichloro-4-pyridyl)-difluoro-methyl]cyclohexyl]amino]-3- oxo-propyl]carbamate Into the solution of Int-3-1 (0.1 g, 339 µmol) in the DCM (1.5 mL) was added 3-((tert- butoxycarbonyl)amino)propanoic acid (0.128 g, 678 µmol), followed by HATU (0.193g, 509 µmol) and DIEA (177 µL, 1.02 mmol). The reaction was heated at 100 °C overnight. After completion, the reaction mixture was diluted with EA, washed with water (50 mL). The organic layer was dried over anhydrous sodium sulfate, concentrated in vacuo to give the crude 004a (150 mg, 94.94% yield), which was used directly in next step. MS observed (ESI⁺) [(M+H)⁺]: 366.8. Step 2: Tert-butyl N-[3-[[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-3- oxo-propyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Compound 004a (150 mg, 322 µmol) and tert-butyl (R)-(5-oxo-1-(4-(piperazin-1- ylsulfonyl)phenyl)pyrrolidin-3-yl)carbamate (100 mg, 236 µmol) were mixed in the DMSO (5 mL). Into the stirring suspension was added DIEA (123 µl, 707 µmol). The reaction mixture was stirred at 120 °C overnight. After the reaction was completed, 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 in vacuo to give crude 004b (150 mg, 176 µmol, 74.5 % yield) as light brown oil. MS obsd. (ESI⁺) [(M+H- BOC)⁺]: 854.3. Step 3: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-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-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide (004B) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Compound 004b (100 mg, 117 µmol) was dissolved in the DCM (5 mL), followed by adding TFA (2 mL, 58.5 µmol). The reaction was stirred at room temperature for 2 hours. After completion, the reaction was concentrated in vacuo to give the crude residue, which was purified by prep-HPLC and then chiral separated by SFC to afford singer isomers: trans Example 004A (eluted first, 28.5 mg, 74.45% yield) as a white solid and cis Example 004B (eluted second, 28 mg, 73.14% yield) as a white solid. The configurations are determined by 2D NMR. Example 004A: MS obsd. (ESI⁺) [(M+H)⁺]: 654.118.¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.97 - 8.08 (m, 1 H), 7.88 - 7.96 (m, 2 H), 7.77 - 7.86 (m, 2 H), 6.76 - 6.82 (m, 1 H), 6.66 - 6.74 (m, 1 H), 4.21 - 4.36 (m, 1 H), 4.00 - 4.16 (m, 1 H), 3.77 - 3.90 (m, 1 H), 3.58 - 3.69 (m, 4 H), 3.03 - 3.14 (m, 1 H), 2.89 - 3.03 (m, 6 H), 2.34 - 2.44 (m, 2 H), 2.08 - 2.24 (m, 1 H), 1.75 - 1.87 (m, 2 H), 1.58 - 1.68 (m, 2 H), 1.02 - 1.34 (m, 6 H).¹⁹F NMR (376 MHz, DMSO-d6) δ ppm - 107.75 - -100.42 (m, 2 F). Example 004B: MS obsd. (ESI⁺) [(M+H)⁺]: 654.118.¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.99 - 8.07 (m, 1 H), 7.87 - 7.98 (m, 2 H), 7.75 - 7.84 (m, 2 H), 6.72 - 6.89 (m, 1 H), 6.60 - 6.75 (m, 1 H), 4.20 - 4.32 (m, 1 H), 4.03 - 4.16 (m, 1 H), 3.74 - 3.91 (m, 2 H), 3.60 - 3.74 (m, 4 H), 3.02 - 3.13 (m, 1 H), 2.85 - 3.02 (m, 4 H), 2.55 - 2.71 (m, 3 H), 2.08 - 2.28 (m, 1 H), 1.61 - 1.80 (m, 2 H), 1.35 - 1.56 (m, 5 H), 0.96 - 1.35 (m, 3 H).¹⁹F NMR (376 MHz, DMSO-d₆) δ ppm - 111.70 - -95.35 (m, 2 F). Example 005: Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-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-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide (005B) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Compound 005A and 005B were prepared in analogy to the preparation of compound 004A and 004B by using (tert-butoxycarbonyl)alanine instead of 3-((tert-butoxycarbonyl)amino)propanoic acid in Step 1. Example 005A (4.3 mg) and Example 005B (5.7 mg) were obtained as white solids. The configurations were determined by 2D NMR. Example 005A: MS obsd. (ESI⁺) [(M+H)⁺]: 654.3.¹H NMR (400 MHz, METHANOL-d4) δ ppm 7.77 - 7.91 (m, 2 H), 7.65 - 7.75 (m, 2 H), 6.51 - 6.63 (m, 2 H), 4.40 - 4.59 (m, 2 H), 3.91 - 4.11 (m, 2 H), 3.77 - 3.90 (m, 1 H), 3.64 - 3.72 (m, 1 H), 3.55 - 3.64 (m, 4 H), 3.46 - 3.55 (m, 1 H), 3.27 - 3.41 (m, 1 H), 2.91 - 3.03 (m, 4 H), 2.74 - 2.87 (m, 1 H), 2.04 - 2.36 (m, 2 H), 1.85 - 1.99 (m, 3 H), 1.64 - 1.79 (m, 2 H), 1.30 - 1.58 (m, 4 H).¹⁹F NMR (376 MHz, METHANOL-d4) δ ppm -109.16 - -99.58 (m, 2 F). Example 005B: MS obsd. (ESI⁺) [(M+H)⁺]: 654.3.¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.76 - 7.89 (m, 2 H), 7.64 - 7.75 (m, 2 H), 6.46 - 6.66 (m, 2 H), 4.40 - 4.61 (m, 1 H), 3.99 - 4.09 (m, 1 H), 3.62 - 3.70 (m, 1 H), 3.54 - 3.61 (m, 4 H), 3.34 - 3.56 (m, 3 H), 2.92 - 3.02 (m, 4 H), 2.72 - 2.86 (m, 1 H), 2.24 - 2.38 (m, 1 H), 2.06 - 2.24 (m, 1 H), 1.90 - 2.01 (m, 1 H), 1.76 - 1.90 (m, 2 H), 1.63 - 1.73 (m, 2 H), 1.30 - 1.55 (m, 2 H), 1.04 - 1.29 (m, 4 H).¹⁹F NMR (METHANOL-d4, 376 MHz) δ -106.71 (br d, 2 F, J=13.6 Hz) Example 006: Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl] 1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 006 Compound 006 was prepared in analogy to the preparation of compound 004A by using -trans- (tert-butoxycarbonylamino)cyclobutanecarboxylic acid instead of 3-((tert- butoxycarbonyl)amino)propanoic acid in Step 1 without chiral separation. Example 006 (76 mg) was obtained as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 680.2.¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.80 - 7.88 (m, 2 H), 7.68 - 7.80 (m, 2 H), 6.52 - 6.64 (m, 2 H), 4.23 - 4.36 (m, 1 H), 4.02 - 4.15 (m, 1 H), 3.78 - 3.96 (m, 2 H), 3.51 - 3.67 (m, 4 H), 3.36 - 3.51 (m, 1 H), 3.01 - 3.14 (m, 2 H), 2.89 - 3.01 (m, 4 H), 2.50 - 2.66 (m, 1 H), 2.34 - 2.50 (m, 2 H), 2.16 - 2.34 (m, 2 H), 1.60 - 2.05 (m, 4 H), 1.31 - 1.54 (m, 3 H), 1.00 - 1.31 (m, 4 H).¹⁹F NMR (376 MHz, METHANOL-d₄) δ ppm -113.39 - -101.55 (m, 2 F). Example 007: Cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide Compound 007 was prepared in analogy to the preparation of compound 004A by using cis-(tert- butoxycarbonylamino)cyclobutanecarboxylic acid instead of 3-((tert- butoxycarbonyl)amino)propanoic acid in Step 1 without chiral separation. Example 007 (47.5 mg) was obtained as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 680.2.¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.89 - 8.00 (m, 2 H), 7.78 - 7.89 (m, 2 H), 6.66 - 6.73 (m, 2 H), 4.36 - 4.47 (m, 1 H), 4.14 - 4.29 (m, 1 H), 3.89 - 4.03 (m, 1 H), 3.65 - 3.79 (m, 5 H), 3.47 - 3.65 (m, 1 H), 3.15 - 3.27 (m, 1 H), 3.00 - 3.15 (m, 5 H), 2.85 - 3.00 (m, 1 H), 2.65 - 2.77 (m, 1 H), 2.50 - 2.65 (m, 2 H), 2.22 - 2.36 (m, 2 H), 1.71 - 2.22 (m, 4 H), 1.43 - 1.63 (m, 3 H), 1.07 - 1.43 (m, 4 H).¹⁹F NMR (376 MHz, METHANOL-d₄) δ ppm -115.37 - -97.60 (m, 2 F). F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 008: Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3- hydroxy-propanamide Compound 008 was prepared in analogy to the preparation of compound 004A by using Int-4-1 instead of Int-3-1, and 3-[tert-butyl(dimethyl)silyl]oxy-2-(9H-fluoren-9- ylmethoxycarbonylamino)propionic acid instead of 3-((tert-butoxycarbonyl)amino)propanoic acid in Step 1. Example 008 (3.15 mg) was obtained as a white solid. MS observed (ESI⁺) [(M+H)⁺]: 670.3.¹H NMR (400 MHz, METHANOL-d4) δ ppm 7.79 - 7.89 (m, 2 H), 7.69 - 7.79 (m, 2 H), 6.54 - 6.63 (m, 2 H), 5.22 - 5.28 (m, 1 H), 4.25 - 4.36 (m, 1 H), 4.01 - 4.14 (m, 1 H), 3.73 - 3.94 (m, 4 H), 3.64 - 3.73 (m, 1 H), 3.52 - 3.64 (m, 4 H), 3.42 - 3.52 (m, 1 H), 3.04 - 3.14 (m, 2 H), 2.92 - 3.04 (m, 4 H), 2.49 - 2.64 (m, 1 H), 2.03 - 2.18 (m, 2 H), 1.86 - 2.03 (m, 2 H), 1.69 - 1.86 (m, 2 H), 1.33 - 1.62 (m, 2 H).¹⁹F NMR (376 MHz, METHANOL-d4) δ ppm -109.45 - -104.93 (m, 1 F). Example 009: Cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]urea Step 1: Cis-tert-butyl N-[2-[[4-[(2,6-dichloro-4-pyridyl)-difluoro- methyl]cyclohexyl]carbamoylamino]ethyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of triphosgene (60.32 mg, 0.203 mmol) in acetonitrile (5 mL) cooled in the ice/water bath was added a solution of tert-butyl N-(2-aminoethyl)carbamate (96.54 µL, 0.610 mmol) and Et3N (51.42 mg, 70.83 uL, 0.508 mmol) in acetonitrile (5 mL) dropwise. Then into the stirring mixture was added a solution of Int-4-1 (60 mg, 0.203 mmol) in acetonitrile (5 mL). The reaction mixture was slowly warmed to room temperature and kept stirring overnight. After completion, the reaction mixture was concentrated in vacuo to give crude product 009a (80 mg, 81.76% yield) as a yellow solid, which was used directly in the next step. MS observed (ESI⁺) [(M+H)⁺]: 481.227. Step 2: Cis-tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-(tert- butoxycarbonylamino)ethylcarbamoylamino]cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Compound 009a (83.72 mg, 0.174 mmol) was dissolved in DMF (10 mL) and tert-butyl N-[(3R)- 5-oxo-1-(4-piperazin-1-ylsulfonylphenyl)pyrrolidin-3-yl]carbamate (73.83 mg, 0.174 mmol) and DIEA (30 µL, 0.174 mmol) were added at room temperature. The mixture was stirred at 125 ^oC overnight. After full conversion of starting material, the reaction mixture was diluted with water F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (50 mL) and extracted with EA (50 mL × 2). The combined organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4 and concentrated in vacuo to give crude 009b (100 mg, 66.13%) as a light yellow solid. MS obsd. (ESI⁺) [(M+H)⁺]: 869.3. Step 3: Cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]urea Compound 009b (100 mg, 0.115 mmol) was dissolved in the DCM (10 mL). Into the stirring solution was added TFA (1 mL). The reaction was stirred at room temperature for 2 hours. After completion, the reaction was concentrated under vacuum to give the crude residue, which was purified by prep-HPLC to give Example 009 (2.5 mg, 2.64%) as a light yellow solid. MS obsd. (ESI⁺) [(M+H)⁺]: 669.245.¹H NMR (400 MHz, METHANOL-d4) δ ppm 7.80 - 7.87 (m, 2 H), 7.67 - 7.78 (m, 2 H), 6.48 - 6.63 (m, 2 H), 4.23 - 4.33 (m, 1 H), 4.00 - 4.14 (m, 1 H), 3.77 - 3.89 (m, 1 H), 3.69 - 3.77 (m, 1 H), 3.52 - 3.66 (m, 4 H), 3.25 - 3.29 (m, 1 H), 3.02 - 3.13 (m, 2 H), 2.94 - 3.02 (m, 4 H), 2.82 - 2.94 (m, 2 H), 2.45 - 2.68 (m, 1 H), 2.07 - 2.20 (m, 1 H), 1.86 - 2.07 (m, 3 H), 1.63 - 1.80 (m, 2 H), 1.40 - 1.55 (m, 5 H), 1.27 - 1.40 (m, 2 H).¹⁹F NMR (376 MHz, METHANOL-d₄) δ ppm -115.65 - -100.71 (m, 2 F). Example 010: Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5,5- dimethyl-5-azoniaspiro[2.4]heptane-6-carboxamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 010 Step 1: (6S)-5,5-dimethyl-5-azoniaspiro[2.4]heptane-6-carboxylic acid (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 at room temperature overnight. After completion, the reaction mixture was concentrated to dryness to give crude 010a (300 mg, 85.79%) as light yellow oil, which was used directly in the next step. Step 2: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[[4-[[(6S)-5,5-dimethyl-5- azoniaspiro[2.4]heptane-6-carbonyl]amino]cyclohexyl]-difluoro-methyl]-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Compound Int-5-1 (150 mg, 0.220 mmol) was dissolved in the DMF (10 mL), and then compound 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 at room temperature overnight. After completion, the reaction mixture was diluted with saturated aqueous NH4Cl-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 and concentrated to dryness to afford the crude 010b (180 mg, 98.14% yield) as light yellow oil, which was used directly in the next step. MS obsd. (ESI⁺) [(M+H)⁺]: 834.6. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 3: Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5,5- dimethyl-5-azoniaspiro[2.4]heptane-6-carboxamide Compound 010b (150 mg, 0.180 mmol) was dissolved in the DCM (5 mL), followed by adding TFA (1.5 mL, 19.47 mmol) at room temperature. The reaction was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated to dryness to afford the crude material, which was purified by prep-HPLC to yield Example 010 (26 mg, 18.71% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 734.4.¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.58 - 8.71 (m, 1 H), 8.27 - 8.41 (m, 2 H), 7.87 - 7.98 (m, 2 H), 7.73 - 7.84 (m, 2 H), 6.76 - 6.85 (m, 1 H), 6.66 - 6.76 (m, 1 H), 4.35 - 4.43 (m, 1 H), 4.21 - 4.33 (m, 1 H), 4.04 - 4.13 (m, 1 H), 3.78 - 3.90 (m, 2 H), 3.63 - 3.70 (m, 4 H), 3.20 - 3.29 (m, 3 H), 3.10 - 3.20 (m, 4 H), 3.02 - 3.10 (m, 1 H), 2.92 - 3.02 (m, 4 H), 2.55 - 2.64 (m, 2 H), 2.25 - 2.45 (m, 2 H), 2.07 - 2.25 (m, 1 H), 1.76 - 1.98 (m, 2 H), 1.57 - 1.76 (m, 2 H), 1.04 - 1.34 (m, 4 H), 0.58 - 0.89 (m, 4 H).¹⁹F NMR (471 MHz, DMSO-d6) δ ppm -126.70 - -92.41 (m, 2 F). Example 011: Trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptane-6-carboxamide (011A) and Trans- (5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5-methyl- 5-azoniaspiro[2.4]heptane-6-carboxamide (011B) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 1: (6S)-5-(2-amino-2-oxo-ethyl)-5-azaspiro[2.4]heptane-6-carboxylic acid Into a solution of (6S)-5-azaspiro[2.4]heptane-6-carboxylic acid (100 mg, 0.708 mmol) in the acetonitrile (5 mL) were added with 3-bromopropionamide (161.5 mg, 1.06 mmol) and K2CO3 (293.71 mg, 2.13 mmol). The reaction 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 × 2). The combined organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, and concentrated to dryness to afford the crude 011a (120 mg, 85.46% yield) as a light yellow solid, which was used directly in the next step. Step 2: Trans-tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[[(6S)-5-(2-amino-2-oxo-ethyl)-5- azaspiro[2.4]heptane-6-carbonyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Into a solution of compound Int-5 (150 mg, 0.220 mmol) in the DMF (10 mL) was added 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 was stirred at room temperature overnight. After completion, the reaction mixture was diluted with saturated NH₄Cl-solution (100 mL). The aqueous layer was extracted with EA (100 mL × 2). The organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous Na2SO4 and concentrated to dryness. The crude material was purified by silica gel chromatography (0-10% MeOH/DCM) to afford 011b (150 mg, 79.13% yield) as a light yellow solid. MS observed (ESI⁺) [(M+H)⁺]: 863.6. 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]heptane-6-carbonyl]amino]cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate Into the solution of compound 011b (150 mg, 0.174 mmol) in the DMF (5 mL) were added iodomethane (21.73 µL, 0.347 mmol) and DIEA (151.71 µL, 0.869 mmol). The reaction was stirred at room temperature overnight. After completion, the reaction mixture was concentrated to dryness to give crude 011c (150 mg, 98.29% yield) as dark brown oil, which was used directly in the next step. MS obsd. (ESI⁺) [(M+H)⁺]: 877.664. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 4: Trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]-5-methyl-5-azoniaspiro[2.4]heptane-6-carboxamide (011A) and Trans- (5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5-methyl- 5-azoniaspiro[2.4]heptane-6-carboxamide (011B) Into a solution of 011c (150 mg, 0.171 mmol) in the DCM (5 mL) was added with TFA (1.5 mL, 19.47 mmol). The reaction was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated to dryness. The crude material was purified by prep-HPLC to yield 011A (eluted first, 8.1 mg, 5.49%) and 011B (eluted second, 10.1 mg, 6.84%) as white solids. Example 011A: MS observed (ESI⁺) [(M+H)⁺]: 777.4.¹H NMR (500 MHz, DMSO-d6) δ ppm 7.92 (d, J=8.70 Hz, 2 H), 7.80 (d, J=8.70 Hz, 2 H), 6.79 (s, 1 H), 6.73 (s, 1 H), 4.57 - 4.75 (m, 1 H), 4.18 - 4.37 (m, 3 H), 4.07 (br d, J=11.14 Hz, 2 H), 3.75 - 3.89 (m, 2 H), 3.66 (br s, 4 H), 3.17 (s, 2 H), 3.08 (br dd, J=17.85, 8.24 Hz, 2 H), 2.98 (br s, 4 H), 2.54 - 2.62 (m, 2 H), 2.40 - 2.49 (m, 1 H), 2.15 (br dd, J=13.73, 4.43 Hz, 2 H), 1.87 (br d, J=10.68 Hz, 1 H), 1.74 - 1.83 (m, 1 H), F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 1.60 - 1.73 (m, 2 H), 1.10 - 1.26 (m, 4 H), 0.72 - 0.91 (m, 4 H).¹⁹F NMR (471 MHz, DMSO-d6) δ ppm -114.06 - -97.82 (m, 2 F). Example 011B: MS observed (ESI⁺) [(M+H)⁺]: 777.5.¹H NMR (500 MHz, DMSO-d₆) δ ppm 7.86 - 7.96 (m, 2 H), 7.77 - 7.84 (m, 2 H), 6.77 - 6.86 (m, 1 H), 6.65 - 6.75 (m, 1 H), 4.57 - 4.66 (m, 1 H), 4.34 - 4.43 (m, 1 H), 4.22 - 4.32 (m, 1 H), 4.11 - 4.22 (m, 1 H), 3.99 - 4.12 (m, 1 H), 3.71 - 3.87 (m, 2 H), 3.61 - 3.71 (m, 4 H), 3.14 - 3.21 (m, 2 H), 3.02 - 3.14 (m, 2 H), 2.86 - 3.02 (m, 4 H), 2.56 - 2.66 (m, 3 H), 2.10 - 2.33 (m, 1 H), 1.77 - 1.91 (m, 3 H), 1.58 - 1.73 (m, 3 H), 1.11 - 1.33 (m, 4 H), 0.81 - 0.92 (m, 2 H), 0.60 - 0.80 (m, 4 H).¹⁹F NMR (471 MHz, DMSO-d6) δ ppm -114.06 - -91.20 (m, 2 F). Example 12 ：(R)-4-amino-1-(4-((4-(6-chloro-4-(difluoro((1r,4r)-4-(3-hydroxyazetidin-1- yl)cyclohexyl)methyl)pyridin-2-yl)piperazin-1-yl)sulfonyl)phenyl)pyrrolidin-2-one Step 1: 1- yl)cyclohexyl)methyl)pyridin-2-yl)piperazin-1-yl)sulfonyl)phenyl)-5-oxopyrrolidin-3- yl)carbamate To a solution of compound Int-2 (100.0 mg, 0.1 mmol) in NMP (3 mL) were added 1,3-dibromo- 2-propanol (32.0 mg, 0.1 mmol) and DIEA (54.0 mg, 0.4 mmol) at 25 ℃. The reaction was stirred F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 at 50 °C for 2 hours. After 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 EtOAc (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by prep-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 give the 12a (74.0 mg, 0.1 mmol, 70.0% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 740.1 Step 2: (R)-4-amino-1-(4-((4-(6-chloro-4-(difluoro((1r,4r)-4-(3-hydroxyazetidin-1- yl)cyclohexyl)methyl)pyridin-2-yl)piperazin-1-yl)sulfonyl)phenyl)pyrrolidin-2-one To a solution of compound 12a (40.0 mg, 0.1 mmol) in DCM (1 mL) was added TFA (1.0 mL, 12.98 mmol) at 25 °C and the mixture was stirred for 2 hours at 25 °C. After the mixture was concentrated under reduced pressure, the residue was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 19% - 49%, 10 min) and lyophilized to give the Example 12 (21.2 mg, 0.1 mmol, 54.5% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 640.3. Example 41: (4R)-4-amino-1-[4-[4-[4-[[4-(azetidin-1-yl)cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 41 Example 41 was prepared in analogy to the preparation of compound Example 12 by using 1,3- dibromopropane instead of 1,3-dibromo-2-propanol in step 1. Example 41 (43.3 mg, 0.1 mmol, 49.0% yield) was obtained as a white solid in form of a TFA salt. MS observed (ESI+) [({³⁵Cl}M+H)⁺]: 623.1. Example 13: Cis-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one Step 1: Tert- -2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate To a solution of compound Int-3 (214.8 mg, 0.5 mmol) and Int-1 (200.0 mg, 0.5 mmol) in DMSO (2 mL) was added DIEA (0.5 mL, 0.51 mmol) at 25 °C and it was heated to 110 °C with stirring for 12 hours. After cooling to room temperature, EtOAc (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 EtOAc (50 mL × 2). The combined organic layer was washed with saturated brine (80 mL × 3), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by prep-TLC (PE/EtOAc = 1/1; TLC: PE/EtOAc = 1/1, Rf = 0.4) to give F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 compound 13a (280.0 mg, 0.4 mmol, 70.6% yield) as a yellow solid. MS observed (ESI⁺) [{³⁵Cl}(M+H)⁺]: 783.4. Step 2: Cis-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one To a solution of compound 13a (250.0 mg, 0.3 mmol) in DCM (3 mL) was added TFA (3 mL) at 25 °C and it was stirred at this temperature for 2 hours. The mixture was concentrated under reduced pressure to give a residue, which was re-dissolved in water (10 mL) and aqueous 5% (w%) NH_3•H₂O (10 mL) was added into the mixture. Solid was formed and the mixture was stirred at 20 °C for 16 hours. The resulting precipitate was filtered. The filter cake was collected and dissolved in water (20 mL), and the suspension was lyophilized to give 13 (153.4 mg, 0.3 mmol, 81.5% yield) as a white solid. MS observed (ESI⁺) [{³⁵Cl}(M+H)⁺]: 583.0. Example 14: Trans-rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one Step 1: Tert-butyl-N-[4-[[2-chloro-6-[4-[4-[rac-(4R)-4-(tert-butoxycarbonylamino)-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of compound Int-2 (200.0 mg, 0.3 mmol) in DCM (5 mL) was added TFA (2.0 mL, 26.0 mmol) at 20 °C and it was stirred at this temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 17% - 47%, 10 min) and lyophilized to give a product (100.0 mg) as HCl salt form. The product was dissolved in water (10 mL) and aqueous 5% (w%) NH3•H2O (10 mL). Solid precipitated out after being stirred for 16 hours at 20 °C and filtered. The filter cake was collected and re-dissolved in water (20 mL). The suspension was lyophilized to give 14 (55.3 mg, 0.1 mmol, 35.4% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 583.1. Example 15: 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]methyl]acetamide Step 1: Tert-butyl N-[2-[[4-[[2-chloro-6-[4-[4-[rac- (4R)-4-(tert-butoxycarbonylamino)-2- oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4-pyridyl]-difluoro- methyl]cyclohexyl]methylamino]-2-oxo-ethyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 15a 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) were added HATU (129.2 mg, 0.3 mmol) and DIEA (70.0 mg, 0.5 mmol) at 25 °C. After the reaction was stirred at 25 °C for 2 hours, the resulting mixture was diluted with H2O (30 mL). Then the mixture was transferred to a separatory funnel and the aqueous layer was extracted with EtOAc (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by prep-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 give 15a (40.0 mg, 0.1 mmol, 59.4% yield) as a white solid. MS observed (ESI⁺) [{³⁵Cl}(M+H)⁺]: 854.3. Step 3: 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]methyl]acetamide To a solution TFA (1.0 mL, 13.0 mmol) at 25 °C and it was stirred at 25 °C for 1 hour. The mixture was concentrated under reduced pressure to afford the crude product, which was purified by prep-HPLC (column: YMC F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 18% - 48%, 10 min) and lyophilized to give Example 15 (18.0 mg, 0.1 mmol, 57.5% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 654.3. Example 16 : 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)amino)- N,N-bis(3-aminopropyl)-N-(carboxymethyl)-4-oxobutan-1-aminium Step 1: To a solution of tert-butyl N-[3-[3- propylamino]propyl]carbamate (2.5 g, 7.7 mmol) in ACN (15 mL) were added benzyl 4-bromobutanoate (2.0 g, 7.7 mmol) and NaHCO3 (1.0 g, 11.5 mmol) at 25 °C and it was stirred at 25 °C for 16 hours. After the mixture was filtrated, the filtrate was concentrated under reduced pressure to afford the crude product, which was purified by prep-HPLC (column: C18 spherical 20 ~ 35 µm, 100 g, mobile phase: [A: F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 water (0.1% TFA) - B: ACN]; B%: 0% - 100%, 15 min) and lyophilized to give 16a (1.8 g, 3.6 mmol, 46.2% yield) as colorless oil. MS obsd. (ESI⁺) [(M+H)⁺]: 508.1. Step 2: 4-(benzyloxy)-N-(2-(tert-butoxy)-2-oxoethyl)-N,N-bis(3-((tert- butoxycarbonyl)amino)propyl)-4-oxobutan-1-aminium bromide To a solution of compound 16a MeCN (3 mL) were added tert-butyl bromoacetate (461.1 mg, 2.4 mmol) and NaHCO₃ (413.7 mg, 4.9 mmol) 25 °C with stirring for 16 hours. After the mixture was filtered, the filtrate was concentrated under reduced pressure to afford the crude product, which was purified by prep-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 give 16b (640.0 mg, 1.0 mmol, 52.2% yield) as colorless oil. MS observed (ESI⁺) [(M+H)⁺]: 622.3. Step 3: N-(2-(tert-butoxy)-2-oxoethyl)-3-((tert-butoxycarbonyl)amino)-N-(3-((tert- butoxycarbonyl)amino)propyl)-N-(3-carboxypropyl)propan-1-aminium bromide To a solution of compound 16b (10 mL) was added wet Pd/C (106.4 mg, 5% wt, containing 50% water) under N₂ atmosphere. The suspension was degassed F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 under vacuum, purged with H₂ for 3 times and stirred at 25 °C for 3 hours under H₂ atmosphere. After being diluted with MeOH (100 mL), the mixture was filtered and the filtrate was concentrated under reduced pressure to afford the crude 16c (550.0 mg, 1.0 mmol, 98.9% yield) as a white solid. The crude product was used in next step directly without further purification. MS observed (ESI⁺) [(M+H)⁺]: 532.4. Step 4: 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin- 1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N- (carboxymethyl)-4-oxobutan-1-aminium (Example 16) 2 and 16c instead of Int-4 and (tert-butoxycarbonyl)glycine respectively in step 1. Example 16 (102.0 mg, 0.1 mmol, 76.3% yield) as a light yellow solid was obtained in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 840.1. Example 17: N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2-(2- aminoethoxy)acetamide

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 17 Example 17 was prepared in analogy to 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, and using T₃P instead of HATU in step 1. Example 17 (12.2 mg, 0.1 mmol, 35.7% yield) as a white solid was obtained in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 684.2. Example 18: 3-amino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2- hydroxypropanamide Example 18 by using Int- 2 and 3-(((benzyloxy)carbonyl)amino)-2-hydroxypropanoic acid instead of Int-4 and (tert- butoxycarbonyl)glycine respectively in step 1. Example 18 (6.0 mg, 0.01 mmol, 22.4% yield) was obtained as a white solid was obtained in form of a hydrochloric acid salt. MS observed (ESI+) [({³⁵Cl}M+H)⁺]: 670.2. Example 19: 2,3-diamino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4- yl)difluoromethyl)cyclohexyl)propanamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 19 by using Int- 2 and 2-(((Benzyloxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid instead of Int-4 and (tert-butoxycarbonyl)glycine respectively in step 1. Example 19 (14.0 mg, 0.02 mmol, 41.8% yield) as a white solid was obtained in form of a hydrochloric acid salt. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 669.2. Example 26: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide Step 1: benzyl 4-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)butanoate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (3.0 g, 9.9 mmol) in ACN (10 mL) were added benzyl 4-bromobutanoate (2.8 g, 10.9 mmol) and potassium carbonate (6.8 g, 49.4 mmol) at 20 °C and it 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 EtOAc (100 mL × 3), washed with saturated brine (100 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by silica gel column (PE: EtOAc = 1: 0 to 0: 1; TLC: PE/EtOAc = 3/1, Rf = 0.5) to give 26a (1.6 g, 3.3 mmol, 99.0% yield) as colorless oil. Step 2: 4-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]butanoic acid To a solution of 26a (800.0 mg, 1.7 mmol) in methanol (8 mL) was added wet Pd/C (170.0 mg, 5% wt, containing 50% water) at 25 °C under N_2. The mixture was degassed under vacuum, purged with H2 for 3 times and stirred at 25 °C for 3 hours under H2 atmosphere (15 psi). After the mixture was diluted with MeOH (100 mL), it was filtered and the filtrate was concentrated in vacuo to give crude 26b (600.0 mg, 1.5 mmol, 92.4% yield) as colorless oil. The crude product was used directly in next step without purification. MS obsd. (ESI⁺) ([M+H]⁺): 390.4. Step 3: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide (Example 26) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-2 and 26b instead of Int-4 and (tert-butoxycarbonyl)glycine respectively in step 1. Example 26 (30.4 mg, 0.03 mmol, 51.0% yield) as yellow solid in form of a TFA salt. MS obsd. (ESI⁺) ([{³⁵Cl}M+H]⁺): 754.1. Example 32: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3- aminopropyl)amino]butanamide Step 1: Benzyl 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 32a To 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 added K₂CO₃ (615.5 mg, 4.5 mmol) at 20 °C and it was heated to 40 °C with stirring for 12 hours. The mixture was poured into water (30 mL) and extracted by EtOAc (50 mL × 3). The combined organic layer was washed with saturate brine (50 mL × 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified by prep-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 give compound 32a (400.0 mg, 0.8 mmol, 52.2% yield) as colorless oil. MS observed (ESI⁺) [(M+H)⁺]: 508.4. Step 2: 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoic acid To a solution of compound 32a (300.0 mg, 0.6 mmol) in methanol (5 mL) was added wet Pd/C (100.0 mg, 5% wt, containing 50% water) at 20 °C. The mixture was degassed under vacuum, purged with H2 for 3 times and it was stirred at 20 °C for 3 hours under H2 atmosphere (15 psi). After being diluted with MeOH (100 mL), the mixture was filtered and the filtrate was concentrated in vacuo to give a crude 32b (200.0 mg, 0.5 mmol, 81.1% yield) as colorless oil. The crude product was used directly in next step without further purification. MS observed (ESI⁺) [(M+H)⁺]: 418.0. Step 3: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Int-2 and 32b instead of Int-4 and (tert-butoxycarbonyl)glycine respectively in step 1. Example 32 (14.7 mg, 0.02 mmol, 38.7% yield) was obtained as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 782.3. Example 31: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N- hydroxy-butanamide Example 31 was prepared in analogy to 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, 26.6% yield) was obtained as a white solid in form of a hydrochloric acid salt. MS observed (ESI) [({³⁵Cl}M+H)⁺]: 798.3. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 21: (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]amino]-4- oxo-butyl]-bis(3-aminopropyl)ammonium Step 1: benzyl 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoate To a solution of tert-butyl N-[3- propylamino]propyl]carbamate (2.5 g, 7.7 mmol) in ACN (15 mL) were added NaHCO₃ (966.5 mg, 11.5 mmol) and benzyl 4- bromobutanoate (2.0 g, 7.7 mmol) at 25 °C and it was stirred at 25 °C for 16 hours. The mixture was filtered and the filtrate was concentrated to give the residue, which was purified by prep-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 give 21a (3.6 g, 7.1 mmol, 92.5% yield) as colorless oil. MS observed (ESI⁺) [(M+H)⁺]: 508.3. Step 2: 4-[bis[3-(tert-butoxycarbonylamino)propyl]amino]butanoic acid F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a suspension of wet Pd/C (20.0 mg, 5% wt, containing 50% water) in methanol (5 mL) was added compound 21a (400.0 mg, 0.8 mmol) under N₂ at 25 °C. The mixture was degassed, purged with H2 for three times and stirred with a H2 balloon at 25 °C for 16 hours. After the mixture was diluted with MeOH (100 mL) and filtered, the filtrate was concentrated to give crude 21b (330.0 mg, 0.8 mmol) as a colorless gum. MS observed (ESI⁺) [(M+H)⁺]: 418.2. Step 3: tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propyl-[4-[[4-[[2-chloro-6-[4-[4-[rac- (4R)-4-(tert-butoxycarbonylamino)-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-4- pyridyl]-difluoro-methyl]cyclohexyl]amino]-4-oxo-butyl]amino]propyl]carbamate To a solution of 21b (300.0 mg, 0.7 mmol) in DMF (2 mL) were added 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) at 0 °C under N₂. After stirred at 0 °C for 0.5 hour, Int-2 (490.9 mg, 0.7 mmol) was added and it was stirred at 25 °C for 1 hour. This mixture was concentrated in vacuo to give a residue, which was purified by prep-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 give 21c (500.0 mg, 0.5 mmol, 61.1% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]:1082.6. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 4: (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]amino]-4-oxo-butyl]-bis[3-(tert- butoxycarbonylamino)propyl]ammonium;iodide To a solution of 21c (150.0 mg, 0.1 mmol) in ACN (2 mL) were added iodoacetamide (153.7 mg, 0.8 mmol) and DIEA (0.1 mL, 0.8 mmol) at 20 °C and it was heated to 40 °C with stirring for 16 hours. The mixture was poured into water (30 mL) and extracted by EtOAc (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 give a residue, which was purified by prep-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 give 21d (80.0 mg, 0.1 mmol, 50.6% yield) as a white solid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 1139.6. Step 5: 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidin- 3-yl]propanamide

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of 21d (80.0 mg, 0.1 mmol) in DCM (2 mL) was added trifluoroacetic acid (1 mL) at 25 °C and it was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo to afford a residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15% - 45%, 10 min) and lyophilized to give Example 21 (46.3 mg, 0.1 mmol, 82.4% yield) as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 839.3. Example 20: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(diaminomethylene)urea Step imidazole-1- carboxamido)cyclohexyl)difluoromethyl)-6-chloropyridin-2-yl)piperazin-1- yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To 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 added 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 by EtOAc (30 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 give crude 20a (110.0 mg, 0.1 mmol, 96.7% yield) as a white solid, which was used in next step directly without further purification. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 777.3. 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-pyrrolidin-3-yl]carbamate To 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 added tert-butyl N-carbamimidoylcarbamate (80.0 mg, 0.5 mmol) at 20 °C, and stirred for 12 hours. The mixture was poured into water (30 mL) and extracted by EtOAc (30 mL × 3). The combined organic phase was washed with saturated brine (50 mL × 3), dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure to F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 give a residue, which was purified by prep-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 give 20b (15.0 mg, 0.02 mmol, 16.8% yield) as a yellow solid. MS obsd. (ESI⁺) [(M+H)⁺]: 868.3. Step 3: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(diaminomethylene)urea To a mL) was added trifluoroacetic acid (0.5 mL, 6.5 mmol) at 20 °C, and stirred for 1 hour. The mixture was concentrated in vacuo to give the crude product, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 17% - 47%, 10 min) and lyophilized to give the 20 (1.5 mg, 0.01 mmol, 12.1% yield) as a yellow solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [(M+H)⁺]: 668.2. Example 22: 3-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoylamino]propyl-bis(3- aminopropyl)-(carboxymethyl)ammonium

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 1: tert-butyl N-[3-[3-(benzyloxycarbonylamino)propyl-[3-(tert- propyl]amino]propyl]carbamate To 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 added NaHCO3 (1.5 g, 18.1 mmol) at 25 °C and it was stirred at 25 °C for 12 hour. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by prep-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 give 22a (1.0 g, 1.9 mmol, 31.7% yield) as yellow oil. MS obsd. (ESI⁺) [(M+H)⁺]: 523.8. Step 2: 3-(benzyloxycarbonylamino)propyl-bis[3-(tert-butoxycarbonylamino)propyl]-(2- tert-butoxy-2-oxo-ethyl)ammonium;bromide

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 22b To a solution of 22a (1.0 g, 1.9 mmol) in DMSO (5 mL) were added NaHCO3 (482.1 mg, 5.7 mmol) and tert-butyl bromoacetate (1.5 g, 7.7 mmol) at 20 °C, and the mixture was stirred at 50 °C for 15 hours. The mixture was diluted with EtOAc (100 mL) and filtered. The filtrate was concentrated to give a residue which was purified by prep-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 give 22b (500.0 mg, 0.7 mmol, 36.4% yield) as yellow oil. MS obsd. (ESI⁺) [(M+H)⁺]: 637.2. Step 3: 3-aminopropyl-bis[3-(tert-butoxycarbonylamino)propyl]-(2-tert-butoxy-2-oxo- ethyl)ammonium;bromide To a suspension of wet Pd(OH)₂/C (0.5 g, 5% wt, containing 50% water) in THF (20 mL) was added 22b (1.0 g, 1.6 mmol) under N2 atmosphere at 25 °C. The suspension was degassed under vacuum and purged with H₂ three times. Then the mixture was 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 afford the crude product 22c (500.0 mg, 1.0 mmol, 63.3% yield) as yellow oil. The 22c was used directly in next step without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 503.5. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 22 was prepared in analogy to the preparation of compound Example 20 by using 22c instead of tert-butyl N-carbamimidoylcarbamate and TEA instead of K₂CO₃ in step 2. Example 22 (14.0 mg, 0.02 mmol, 22.5% yield) was obtained as yellow solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 855.5. Example 23: 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea Example 23 was prepared in analogy to the preparation of compound Example 20 by using tert- butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate instead of tert-butyl N- carbamimidoylcarbamate and DIEA instead of K2CO3 in step 2. in step 2. Example 23 ((16.0 mg, 0.02 mmol, 40.0% yield) as a white powder was obtained in form of a hydrochloric acid. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 740.4. Example 34: (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4- hydroxy-pyrrolidine-1-carboxamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 34 was prepared in analogy to the preparation of compound Example 20 by using tert- butyl N-[(3R,4S)-4-hydroxypyrrolidin-3-yl]carbamate instead of tert-butyl N- carbamimidoylcarbamate and DIEA instead of K2CO3 in step 2. Example 34 (41.6 mg, 0.1 mmol, 53.3% yield) was obtained as white solid in form of a hydorchloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 711.3. Example 35: 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-3-(2- (bis(2-aminoethyl)amino)ethyl)urea Step 1: tert-butyl N-[2-[2-(benzyloxycarbonylamino)ethyl-[2-(tert- butoxycarbonylamino)ethyl]amino]ethyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (1.0 g, 3.3 mmol) in ACN (10 mL) were added benzyl N-(2-bromoethyl)carbamate (1.0 g, 4.0 mmol) and NaHCO₃ (830.6 mg, 9.9 mmol) at 20 °C. After the mixture was stirred at 20 °C for 15 hours, it was diluted with EtOAc (100 mL) and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column (PE/EtOAc = 10/1 to 1/1; TLC: PE/EtOAc = 3/1, Rf = 0.4) to give 35a (1.0 g, 2.1 mmol, 63.1% yield) as colorless oil. MS obsd. (ESI⁺) [(M+H)⁺]: 481.1. Step 2: tert-butyl N-[2-[2-aminoethyl-[2-(tert- butoxycarbonylamino)ethyl]amino]ethyl]carbamate To a solution of 35a (300.0 mg, 0.6 mmol) (2 mL) and isopropanol (2 mL) was added dry Pd/C (100.0 mg, 10% wt) at 20 °C under N₂ atmosphere. The suspension was degassed under vacuum and purged with H2 three times. Then the mixture was 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 give crude 35b (200.0 mg, 0.6 mmol, 92.5% yield) as yellow oil, which was used in next step directly without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 347.1. Step 3: 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin- 1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2- aminoethyl)amino)ethyl)urea

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 35 Example 35 was prepared in analogy to the preparation of compound Example 20 by using 35b instead of tert-butyl N-carbamimidoylcarbamate and DIEA instead of K₂CO₃ in step 2. Example 35 (34.0 mg, 0.1 mmol, 50.0% yield) as white solid was obtained in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 755.2. Example 29: (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]morpholine-4-carboxamide trans Step 1: Tert-butyl N-(oxiran-2-ylmethyl)carbamate To a solution of tert-butyl N-allylcarbamate (20.0 g, 127.2 mmol) in DCM (100 mL) was added m-CPBA (32.8 g, 152.7 mmol) at 20 °C and it was stirred at 20 °C for 1 hour. The reaction was quenched with aqueous Na2SO3 (100 mL) and the mixture was stirred at 20 °C for 1 hour. After it was extracted with EtOAc (200 mL × 3), the combined organic layer was washed with saturated aqueous NaHCO3 (50 mL × 3), saturated brine (50 mL × 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give crude 29a (15.0 g, 86.6 mmol, 68.1% yield) as yellow oil. The crude product was used directly in next step without further purification. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 2: Tert-butyl N-[3-(benzylamino)-2-hydroxy-propyl]carbamate 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 give a residue, which was purified by prep-HPLC (column: C18 spherical 20 ~ 35 µm, 330 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0% - 70%, 20 min) and lyophilized to give compound 29b (12.0 g, 42.8 mmol, 41.2% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 281.1. Step 3: Tert-butyl N-[3-[benzyl-(2-chloro-2-cyano-ethyl)amino]-2-hydroxy- propyl]carbamate To a solution of compound 29b (15.0 g, 53.5 mmol) in THF (200 mL) was added 2- chloroacrylonitrile (4.9 g, 56.2 mmol) at 0 °C and the mixture was allowed to warm to 20 °C with stirring for 16 hours. The mixture was poured into water (300 mL) and extracted with EtOAc (300 mL × 3). The combined organic layer was washed with saturated brine (300 mL × 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give crude 29c (12.0 g, 32.6 mmol, 61.0% yield) as light yellow oil, which was used directly in next step without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 368.1. Step 4: Tert-butyl N-[[rac-(2S,6R)-4-benzyl-6-cyano-morpholin-2-yl]methyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 trans cis 29d-1 To a solution of compound 29c (15.0 g, 40.8 mmol) in THF (100 mL) was added solution of potassium tert-butoxide in THF (1.0 M, 41.0 mL, 41.0 mmol) at 0 °C and it was warmed to 20 °C with stirring for 2 hours. The mixture was poured into water (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layer was washed with brine (50 mL × 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (column: C18 spherical 20 ~ 35 µm, 330 g, mobile phase: [A: water (0.1% TFA) - B: ACN]; B%: 0% - 100%, 30 min) and lyophilized to give 29d-1 (5.2 g, 15.7 mmol, 27.5% yield) and 29d-2 (4.1 g, 12.4 mmol, 21.7% yield) as light yellow oil. MS obsd. (ESI⁺) [(M+H)⁺]: 332.1. Step 5: Tert-butyl N-[[6-(aminomethyl)-4-benzyl-morpholin-2-yl]methyl]carbamate trans To a solution of compound 29d-1 (200.0 mg, 0.6 mmol) in THF (3 mL) was added solution of BH₃•Me₂S (10.0 M, 0.5 mL, 5.0 mmol) at 0 °C and it was stirred at 20 °C for 2 hours. The mixture was quenched with MeOH (50 mL) carefully at 0 °C and it was concentrated under reduced pressure to give crude 29e (150.0 mg, 0.5 mmol, 74.1% yield) as colorless oil, which was used directly in next step without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 336.1. Step 6: Tert-butyl N-[[4-benzyl-6-[(tert-butoxycarbonylamino)methyl]morpholin-2- yl]methyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 trans To a solution of DIEA (0.3 mL) and di-tert-butyl dicarbonate (100.0 mg, 0.5 mmol) in THF (1 mL) was added compound 29e (150.0 mg, 0.5 mmol) at 20 °C and it was stirred at this temperature for 12 hours. The reaction mixture were poured into EtOAc (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 purified by silica gel column (PE/EtOAc = 50/1 to 2/1; TLC: PE/EtOAc = 3/1, Rf = 0.4) to give compound 29f (150.0 mg, 0.3 mmol, 77.0% yield) as colorless oil. MS obsd. (ESI⁺) [(M+H)⁺]: 436.1. Step 7: Tert-butyl (((2R,6R)-morpholine-2,6-diyl)bis(methylene))dicarbamate trans To a solution of compound 29f (5 mL) was added wet Pd/C (50.0 mg, 5% wt, containing 50% water) at 20 °C under N₂ atmosphere. The suspension was degassed under vacuum and purged with H2 three times and it was 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 afford crude compound 29g (50.0 mg, 0.2 mmol, 36.3% yield) as colorless oil, which was used directly in next step without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 346.4. Step 8: (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]morpholine-4-carboxamide (Example 29) F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 trans Example 29 was prepared in analogy to the preparation of compound Example 20 by using 29g instead of tert-butyl N-carbamimidoylcarbamate and DIEA instead of K2CO3 in step 2. Example 29 (26.1 mg, 0.03 mmol, 56.3% yield) was obtained as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 754.5. Example 24: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(3-aminopropyl)guanidine Step 1: tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(imidazole-1- carboximidoylamino)cyclohexyl]methyl]-2-pyridyl]piperazin-1- yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 24a To a solution of Int-2 (60.0 mg, 0.1 mmol) in DMF (0.4 mL) was added imidazol-1-yl(1,2,4- triazol-4-yl)methanimine (85.4 mg, 0.5 mmol) under N₂ at 25 °C and it was stirred at 45 °C for 12 hours. After cooling to room temperature, the mixture was directly purified by prep-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 give 24a (50.0 mg, 0.1 mmol, 34.5% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 776.3. Step 2: tert-butyl N-[3-[[N-[4-[[2-[4-[4-[(4R)-4-(tert-butoxycarbonylamino)-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamimidoyl]amino]propyl]carbamate To a solution of 24a (50.0 mg, 0.1 mmol) in DMF (0.2 mL) was added N-Boc-1,3-diaminopropane (157.1 mg, 0.9 mmol) under N2 at 25 °C and it was heated to 80 °C with stirring for 3 hours. After cooling to room temperature, the mixture was directly purified by prep-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 give 24b (30.0 mg, 0.03 mmol, 37.7% yield) as a yellow solid. MS obsd. (ESI⁺) [{³⁵Cl}M+H]⁺]: 882.5. Step 3: 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(3-aminopropyl)guanidine F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of 24b (25.0 mg, 0.03 mmol) in DCM (1 mL) was added TFA (1 mL) at 25 °C and it was stirred at 25 °C for 1 hour. After the mixture was concentrated, the residue was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 14% - 44%, 10 min) and lyophilized to give 24 (5.6 mg, 0.01 mmol, 25.7% yield) as a yellow solid in form of a hydrochloric acid salt. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 682.3. Example 25: N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]-2,2,2-trifluoro-acetamide Step 1: tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-[[(2,2,2- trifluoroacetyl)amino]methyl]cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]- 5-oxo-pyrrolidin-3-yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of Int-4 (70.0 mg, 0.1 mmol) in DCM (0.7 mL) were added trifluoroacetic anhydride (63.0 mg, 0.3 mmol) and DIEA (129.5 mg, 1.0 mmol) at 25 °C. After the mixture was stirred at 25 °C for 2 hours, it was quenched by water (30 mL), extracted with EtOAc (60 mL × 3). The combined organic layer was washed with saturated brine (60 mL), dried over anhydrous Na2SO4 and filtrate. The filtrate was concentrated in vacuo to afford a residue, which was purified by prep- TLC (PE/EtOAc = 1/2; TLC: PE/EtOAc = 1/2, R_f = 0.5) to give 25a (50.0 mg, 0.1 mmol, 62.8% yield) as a light yellow solid. MS obsd. (ESI+) ([{³⁵Cl}M+H]⁺): 793.3. Step 2: N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]- 6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]-2,2,2-trifluoro-acetamide To a solution of 25a (35.0 mg, 0.04 mmol) in DCM (1 mL) was added TFA (2.0 mL) 25 °C and it was stirred at 25 °C for 1 hour. After the mixture was concentrated in vacuo, the residue was purified by prep-HPLC (column: Phenomenex Synergi Polar-RP 100 × 25 mm × 4 um; mobile phase: [A: water (0.1% TFA) - ACN]; B%: 20% - 60%, 10 min)) and lyophilized to give 25 (9.2 mg, 0.01 mmol, 24.0% yield) as a white solid in form of a TFA salt. MS observed (ESI+) ([{³⁵Cl}M+H]⁺): 693.2. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 36: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate Step 1: di-tert-butyl (((2-((tert-butyldimethylsilyl)oxy)ethyl)azanediyl)bis(ethane-2,1- diyl))dicarbamate To a solution of tert-butyl N-[2-[2-(tert-butoxycarbonylamino)ethylamino]ethyl]carbamate (7.2 g, 23.7 mmol) in acetonitrile (30 mL) were added (2-bromoethoxy)-tert-btyldimethylsilane (5.7 g, 23.7 mmol) and NaHCO3 (6.0 g, 71.2 mmol) at 25 °C. After the mixture was stirred at 45 °C for 72 hours, the mixture was diluted with EtOAc (100 mL) and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by silica gel (EtOAc/MeOH = 1:0 to 10/1; TLC: EtOAc/MeOH = 10/1, Rf = 0.3) to give 36a (2.7 g, 5.9 mmol, 24.6% yield) as colorless oil. (ESI+) [(M+H)+]: 462.1. Step 2: di-tert-butyl (((2-hydroxyethyl)azanediyl)bis(ethane-2,1-diyl))dicarbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of 36a (6.6 g, 14.3 mmol) in water (7.0 mL) and THF (30 mL) was added AcOH (13 mL) at 20 °C and stirred for 12 hours. The mixture was purified by prep-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, 54.4% yield) as colorless oil. MS obsd. (ESI+) [(M+H)+]: 348.3. Step 3: 2-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]ethyl (4-nitrophenyl) carbonate 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) was added N,N-diisopropylethylamine (558.0 mg, 4.3 mmol) at 25°C and it was warmed to 40 °C with stirring for 48 hours. The mixture was purified by prep-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, 36.6% yield) as colorless oil. MS obsd. (ESI+) [(M+H)⁺]:513.3. Step 4: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-[bis[2-(tert- butoxycarbonylamino)ethyl]amino]ethoxycarbonylamino]cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of Int.2 (200.0 mg, 0.3 mmol) and 36c (300.1 mg, 0.6 mmol) in THF (15 mL) was added N,N-diisopropylethylamine (0.5 mL, 2.9 mmol) at 25 °C and it was heated to 60 °C with stirring for 48 hours. After the mixture was concentrated in vacuo, the residue was purified by prep- 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 give 36d (80.0 mg, 0.1 mmol, 25.9% yield) as a white solid. (ESI⁺) [({³⁵Cl}M+H)⁺]: 1056.7. Step 5: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate To a solution of 36d (60.0 mg, 0.1 mmol) in DCM (2 mL) was added TFA (1 mL) at 25 °C and it was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo to give a residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 14% - 44%, 10 min) and lyophilized to give 36 (17.8 mg, 0.02 mmol, 40.2% yield) as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 756.1. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 27: 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate Example 27 was prepared in analogy to the preparation of compound 36 using di-tert-butyl (azanediylbis(propane-3,1-diyl))dicarbamate instead of di-tert-butyl (azanediylbis(ethane-2,1- diyl))dicarbamate in Step 1. Example 27 (6.3 mg, 0.01 mmol, 43.0% yield) as a white powder was obtained in form of a TFA salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 784.4. Example 28: 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoyloxy]ethyl-bis(3- aminopropyl)-(carboxymethyl)ammonium Step [2-[tert- butyl(dimethyl)silyl]oxyethyl]amino]propyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propylamino]propyl]carbamate (1.0 g, 3.0 mmol) in acetonitrile (10 mL) were added (2-bromoethoxy)-tert-butyldimethylsilane (721.8 mg, 3.0 mmol) and NaHCO3 (760.4 mg, 9.1 mmol) at 45 °C. After the mixture was stirred at 45 °C for 72 hours, it was filtered and the filtrate was concentrated in vacuo to give residue, which was purified by silica gel column (EtOAc/MeOH = 1/0 to 10/1; TLC: EtOAc/MeOH = 10/1, Rf = 0.3) to give 28a (950.0 mg, 1.9 mmol, 64.3% yield) as colorless oil. MS obsd. (ESI⁺) [(M+H)⁺]: 490.4. Step 2: 2-[bis(3-aminopropyl)amino]ethanol The 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 in vacuo to give crude 28b (216.0 mg, 1.0 mmol, 99.9% yield) as colorless oil, which was used in next step directly without further purification. MS obsd. (ESI⁺) [(M+H)⁺]: 176.0 Step 3: tert-butyl N-[3-[3-(tert-butoxycarbonylamino)propyl-(2- hydroxyethyl)amino]propyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of 28b (216.0 mg, 1.0 mmol) in methanol (15 mL) were added sodium hydrogen carbonate (342.8 mg, 4.1 mmol) and di-t-butyldicarbonate (445.3 mg, 2.0 mmol) at 25 °C. After the mixture was stirred at 25 °C for 1 hour, it was diluted with EtOAc (50 mL) and filtered. The filtrate was concentrated in vacuo to give the residue, which was purified by silica gel column (EtOAc/MeOH = 1/0 to 10/ 1; TLC: EtOAc/MeOH = 10/1, Rf = 0.3) to give 28c (180.0 mg, 0.5 mmol, 47.0% yield) as yellow oil. MS obsd. (ESI⁺) [(M+H)⁺]: 376.1. Step 4: tert-butyl N-[(3R)-1-[4-[4-[4-[[4-[2-[bis[3-(tert- butoxycarbonylamino)propyl]amino]ethoxycarbonylamino]cyclohexyl]-difluoro-methyl]-6- chloro-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate To a solution of 28c (50.0 mg, 0.1 mmol) in THF (2 mL) were added N,N-diisopropylethylamine (0.1 mL, 0.6 mmol) and 4-nitrophenyl chloroformate (30.0 mg, 0.2 mmol) at 0 °C under N2. After the mixture was stirred 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, the mixture was stirred at 40 °C for 48 hours. The mixture was concentrated in vacuo to give a residue. After the residue was dissolved with DMF (2 mL), it was purified by prep-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 give 28d (50.0 mg, 0.1 mmol, 34.6% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 1084.7. 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-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin- 1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)carbamoyl)oxy)ethyl)-N-(3-((tert- butoxycarbonyl)amino)propyl)propan-1-aminium F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 The mixture of 28d (45.0 mg, 0.04 mmol), tert-butyl bromoacetate (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 with FA to pH = 5, follow by purification by prep-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 give 28e (50.0 mg, 0.04 mmol, 87.8% yield) as a yellow solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 1198.5 Step 6: 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)- (carboxymethyl)ammonium (Example 28) To a solution of 28e (40.0 mg, 0.03 mmol) in DCM (2 mL) was added TFA (2 mL) at 25 °C under N2. After the mixture was stirred at 25 °C for 1 hour, it was concentrated in vacuo to give a residue, which was purified by pre-HPLC prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15% - 45%, 10 min) and lyophilized to give 28 (15.0 mg, 0.02 mmol, 47.9% yield) as a brown solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 842.3. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 30: N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide Step 1: To a mixture of methyltriphenylphosphonium bromide (7.5 g, 20.9 mmol) in THF (70 mL) was added dropwise n-butyllithium solution (2.5 M, 7.5 mL, 18.8 mmol) at 0 °C. After a solution of Int-2c (3.9 g, 10.5 mmol) in THF (70 mL) was added, it was continued to be stirred at 0 °C for 2 hours. The reaction mixture was poured into 100 mL of saturated aqueous NH₄Cl and extracted with EtOAc (100 mL × 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered. The filtrate was concentrated in vacuo to afford a residue, which was purified by silica gel column (PE/EtOAc = 50/1 to 2/1; TLC: PE/EtOAc = 2/1, Rf = 0.4) to give 30a (2.9 g, 7.8 mmol, 74.8% yield) as a yellow solid. MS obsd. (ESI⁺) [({³⁵Cl}M- C4H8+H)⁺]:315.0. Step 2: tert-butyl N-[4-[1-(2,6-dichloro-4-pyridyl)cyclopropyl]cyclohexyl]carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Trimethylsulfoxoniumiodide (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 in one portion at 0 °C. Then 30a (200.0 mg, 0.5 mmol) was added to the solution at 0 °C and it was continued to be stirred at 25 °C for 2 hours. The mixture was poured into 20 mL of saturated aqueous NH4Cl and extracted with EtOAc (50 mL × 3). The organic layer was combined, washed with saturated brine (50 mL × 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by prep-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 give 30b (160.0 mg, 0.4 mmol, 77.1% yield) as a yellow solid. MS obsd. (ESI+) [({³⁵Cl}M-C4H8+H)⁺]:329.0. Step 3: tert-butyl N-[4-[1-(2,6-dichloro-4-pyridyl)cyclopropyl]cyclohexyl]carbamate To a solution of 30b (200.0 mg, 0.5 mmol) in DCM (2 mL) was added TFA (2 mL) in one portion at 25 °C and it was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo to give crude 30c (160.0 mg, 0.6 mmol) as a white solid, which was used in next step directly without further purification. MS obsd. (ESI+):295.2 [({³⁵Cl}M+H)+]. Step 4: tert-butyl N-(4-formylcyclohexyl)carbamate F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 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) was added sodium hydrogen carbonate (1.4 g, 16.5 mmol) at 25 °C in one portion. After the mixture was stirred at 25 °C for 15 hours, it was diluted with EtOAc (200 mL) and filtered. The filtrate was concentrated in vacuo to give a residue, which was purified by prep-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 give 30d (380.0 mg, 0.8 mmol, 48.1% yield) as yellow oil. MS obsd. (ESI+) [(M+H)⁺]:480.2. Step 5: 4-[bis[2-(tert-butoxycarbonylamino)ethyl]amino]butanoic acid To a solution of 30d (180.0 mg, 0.4 mmol) in methanol (20 mL) was added wet Pd/C (50 mg, 5% wt, containing 50% water) at 20 °C under N₂ atmosphere. The suspension was degassed under vacuum and purged with H2 three times. Then the reaction was stirred at 40 °C for 16 hours under H2 (15 psi). The mixture was diluted with methanol (200 mL) and filtered. The filtrate was concentrated under reduced pressure to afford the crude product, which was purified by prep-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 give 30e (120.0 mg, 0.3 mmol, 82.1% yield) as colorless oil. MS obsd. (ESI+) [(M+H)⁺]:390.2. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 6: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[1,4-dioxaspiro[4.5]decan-8- yl(difluoro)methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3- yl]carbamate To a solution of 30e (98.3 mg, 0.3 mmol) and 30b (60.0 mg, 0.2 mmol) in DMF (2 mL) were added 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) at 0 °C in one portion. After the mixture was stirred at 25 °C for 2 hours, it was concentrated in vacuo to give a residue, which was purified by prep-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 give 30f (60.0 mg, 0.1 mmol, 43.4% yield) as a yellow solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 656.3. 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-pyrrolidin-3-yl]carbamate

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 30g To a solution of 30f (200.0 mg, 0.3 mmol) in DMSO (1 mL) were added Int-1 (129.3 mg, 0.3 mmol) and N,N-diisopropylethylamine (116.0 mg, 0.9 mmol) at 25 °C and it was heated to 100 °C with stirring for 16 hours. After cooling to 25 °C, the mixture was concentrated in vacuo to give a residue, which was purified by prep-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 give 30g (80.0 mg, 0.1 mmol, 25.1% yield) as a yellow solid. MS obsd. (ESI⁺) [({³⁵Cl}M-Boc+H)⁺]: 944.4. Step 8: Tert-butyl N-[(3R)-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]-5-oxo-pyrrolidin-3- yl]carbamate To a solution of 30g (60.0 mg, 0.1 mmol) in DCM (2 mL) was added TFA (3 mL) at 25 °C in one portion and it was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo to afford a residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15% - 45%, 10 min) and lyophilized to give 30 (8.0 mg, 0.01 mmol, 17.4% yield) as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [({³⁵Cl}M+H)⁺]: 744.3. Example 33: (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxy- propyl)amino]cyclohexyl]-difluoro-methyl]-2-pyridyl]piperazin-1- yl]sulfonylphenyl]pyrrolidin-2-one F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 1: 3-[tert-butyl(dimethyl)silyl]oxy-2,2-difluoro-propan-1-ol To a solution of 2,2- mmol) in DMF (20 mL) were added imidazole (1.2 g, 17.8 mmol) and tert-butyldimethylchlorosilane (1.6 g, 10.8 mmol) at 0 °C. After the mixture was stirred at 0 °C for 4 hours, it was diluted with H2O (30 mL) and transferred to a separatory funnel. The aqueous layer was extracted with EtOAc (80 mL × 3). The combined organic layer was washed with brine (50 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column (PE/EtOAc= 1/0 to 5/1; TLC: PE/EtOAc = 5/1, Rf = 0.4) to give 33a (2.0 g, 98.8% yield) as colorless oil. The 33a was used in next step directly without characterization analysis. Step 2: 3-[tert-butyl(dimethyl)silyl]oxy-2,2-difluoro-propanal To a solution of 33a (300.0 mg, 1.3 mmol) in DCM (5 mL) was added DMP (618.4 mg, 1.5 mmol) 0 °C and it was stirred at 0 °C for 2 hours. After the mixture was quenched by slow addition of saturated K2CO3 solution (20 mL), it was transferred to a separatory funnel and the aqueous layer was extracted with EtOAc (40 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 2), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 to afford the crude product, which was purified by silica gel column (PE/EtOAc= 1/0 to 5/1; TLC: PE/EtOAc = 5/1, Rf = 0.5) to give 33b (250.0 mg, 1.1 mmol, 84.1% yield) as colorless oil. The 33b was used in next step directly without characterization analysis. 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]piperazin-1- yl]sulfonylphenyl]-5-oxo-pyrrolidin-3-yl]carbamate To a solution of 33b (60.0 mg, 0.2 mmol) and Int-2 (201.0 mg, 0.3 mmol) in DCE (1 mL) were added acetic acid (32.1 mg, 0.5 mmol) and sodium triacetoxyborohydride (283.4 mg, 1.3 mmol) in one portion at 25 °C under N₂. After the mixture was stirred at 25 °C for 16 hours, it was diluted with EtOAc (100 mL) and water (20 mL). The organic layer was separated and then washed with saturated brine (30 mL × 2), dried over anhydrous Na2SO4, filtered. The filtrate was concentrated under reduced pressure to give crude 33c (50.0 mg, 0.01 mmol, 48.4% yield) as a white solid, which was used in next step directly without further purification. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 891.2. Step 4: (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxy- propyl)amino]cyclohexyl]-difluoro-methyl]-2-pyridyl]piperazin-1- yl]sulfonylphenyl]pyrrolidin-2-one To a solution of 33c (30.0 mg, 0.03 mmol) in DCM (1 mL) was added TFA (1 mL) at 25 °C and it was stirred at 25 °C for 2 hours. The mixture was concentrated under reduced pressure to afford F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 the crude product, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 20% - 50%, 10 min) and lyophilized to give 33 (7.5 mg, 0.01 mmol, 28.4% yield) as a white solid. MS obsd. (ESI⁺) [({³⁵Cl}M+H)⁺]: 677.2. Example 37: (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-methyl- pyrimidin-2-yl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one Step 1: (4R)-4- methyl-pyrimidin- 2-yl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one The Int-5j (35.0 at 25 °C and it was heated to 60 °C with stirring 16 hours. The mixture was concentrated under reduced pressure to give a residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 12% - 42%, 10 min) and lyophilized to give 37 (19.4 mg, 0.03 mmol, 75.6% yield) as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [(M+H)⁺]:564.4. Example 38: N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-methyl-pyrimidin-4-yl]-difluoro-methyl]cyclohexyl]-4-[bis(3- aminopropyl)amino]butanamide F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example 38 was prepared in analogy to 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 step1. Example 38 (3.5 mg, 0.01 mmol, 7.3% yield) was obtained as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [(M+H)⁺]: 763.6. Example 40: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo- pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate Step 1: 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate;trihydrochloride F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 To a solution of Int-7 (50.0 mg, 0.1 mmol) in DCM (2 mL) was added trifluoroacetic acid (1.00 mL, 12.98 mmol) at 25 °C and stirred for 1 hour. The mixture was concentrated in vacuo to give residue, which was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 15% - 45%, 10 min) and lyophilized to give Example 40 (32.4 mg, 0.1 mmol, 70.7% yield) as a white solid in form of a hydrochloric acid salt. MS observed. (ESI⁺) [({³⁵Cl}M+H)⁺]: 597.3. Example 39 : (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)pyrrolidin-2-one Step 1: tert-butyl ((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate To a solution of Int-5 (500.0 mg, 0.8 mmol) in isopropanol (2 mL) were added 1,1,1-trifluoro-2,3- epoxypropane (84.4 mg, 0.8 mmol) and DIEA (294.1 mg, 2.3 mmol) at 25 ℃ and it was heated to 70 °C with stirring 2 hours. After the mixture was quenched by addition of saturated aqueous ammonium chloride (20 mL), it was transferred to a separatory funnel and the aqueous layer was F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 extracted with EtOAc (60 mL × 3). The combined organic layer was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column (EtOAc/MeOH = 1/0 to 10/1, 1‰ NH₃•H₂O as additive; TLC: EtOAc/MeOH = 10/1, R_f = 0.4) to give the 39a (400.0 mg, 0.5 mmol, 67.8% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 776.2 Step 2: (4R)-4-amino-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)pyrrolidin-2-one To a solution of 39a (400.0 mg, 0.5 mmol) in DCM (5 mL) was added TFA (3 mL) at 25 °C and it was stirred at 25 °C for 2 hours. After the mixture was concentrated under reduced pressure, the crude Example 39 (304.0 mg, 0.5 mmol, 88.0% yield) was obtained as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 676.2. Example 42: tert-butyl ((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate Step 1: tert-butyl ((1r,4S)-4-((2-(4-((4-((R)-4-((tert-butoxycarbonyl)amino)-2-oxopyrrolidin- 1-yl)phenyl)sulfonyl)piperazin-1-yl)-6-methylpyrimidin-4- F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 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-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-methylpyrimidin-4-yl)difluoromethyl)cyclohexyl)((R)- 3,3,3-trifluoro-2-hydroxypropyl)carbamate (42b) To a solution of 39 (300.0 mg, 0.4 mmol) in DCM (5 mL) were added di-t-butyldicarbonate (210.0 mg, 1.0 mmol) and DIEA (148.9 mg, 1.2 mmol) at 25 °C and it was stirred at 25 °C for 2 hours. After the resulting mixture was poured into water (50 mL) and transferred to a separatory funnel, the aqueous layer was extracted with EtOAc (80 mL × 3). The combined organic layer was washed with brine (50 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to afford the crude product, which was purified by silica gel column (PE/EtOAc= 10/1 to 0/1; TLC: PE/EtOAc = 1/1, Rf = 0.4) and then separated by prep-SFC (column: Chiralpak IG 25 × 30 mm I.D., 10 um, mobile phase: 40% IPA (0.1% NH3•H2O) in Supercritical CO2) to give the 42a (eluted first, 140.0 mg, 0.16 mmol, 41.54% yield) as a white solid MS obsd. (ESI⁺) [(M+H)⁺]: 876.2 and 42b (eluted second, 160.0 mg, 0.18 mmol, 47.47% yield) as a white solid. MS obsd. (ESI⁺) [(M+H)⁺]: 876.2. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Step 2: (R)-4-amino-1-(4-((4-(4-(difluoro((1R,4r)-4-(((R)-3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)pyrrolidin-2-one To a solution of 42b (120.0 mg, 0.1 mmol) in DCM (2 mL) was added TFA (2 mL) at 25 °C and it was stirred at 25 °C for 2 hours. After the reaction was concentrated under reduced pressure, the residue was purified by prep-HPLC (column: YMC Triart C18150 × 25 mm × 5 um, mobile phase: [A: water (0.1% HCl) - B: ACN]; B%: 18% - 48%, 10 min) and lyophilized to give compound 42 (71.2 mg, 0.1 mmol, 73.6% yield) as a white solid in form of a hydrochloric acid salt. MS observed (ESI⁺) [(M+H)⁺]: 676.3. Example 43: Minimal Inhibitory Concentration Protocol (MIC) Assay: The antibacterial activity of the compounds of the present invention was evaluated against the commonly used quality control strain Escherichia coli ATCC 25922 and the rifampin-resistant mutant strain Klebsiella pneumonia ATCC 43816. Both were originally derived from human clinical samples and are available from ATCC (American Type Culture Collection). The in vitro potency of compounds to inhibit Escherichia coli (ATCC 25922) and Klebsiella pneumonia (ATCC 43816) growth was assessed by the MIC (Minimal Inhibitory Concentration) broth dilution method. Specifically compound dilutions were prepared from 10 mM DMSO stock solutions as follows: i) serial 2-fold dilution in 20 µL DMSO were prepared in a master plate (Greiner, Cat No: 651201), ii) 180 µL sterile distilled water was added to each aliquot and iii) 10 µL diluted compounds were transferred into a new assay plate (Costar, 3599). Vials of each test microorganisms were maintained frozen in the vapor phase of a liquid nitrogen freezer. Single-use frozen vials of the two strains Escherichia coli ATCC 25922 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (KWIKSTIK, 0335K) and Klebsiella pneumonia ATCC 43816, with predetermined CFU/mL, were taken out from the freezer, thawed at room temperature, and diluted in Cation-Adjusted Mueller Hinton Broth (CAMHB) to achieve a final inoculum of 5 × 10⁵ CFU/ mL.90 µL bacteria containing broth was dispensed to the assay plate containing the pre-dispensed compound dilutions and mixed by pipetting 5 times. Then the assay plates were incubated for 20 hours at 35 °C in ambient air. Following incubation, the MIC (µg/mL), the lowest concentration of drug that inhibits visible growth of the microorganism was read with the help of a magnification mirror and recorded. Compound F-1, disclosed in the literature (Bioorganic Chemistry 102 (2020) 104055) and reported having LPS synthesis pathway inhibition activity and displaying antibiotic activity against efflux-deficient E. coli strains, was chosen as reference compound in present invention. Table 1: MIC values of the compounds of this invention against E. coli and K. pneumonia MIC (μg/mL) MIC (μg/mL) Example Escherichia Klebsiella Example Escherichia Klebsiella No. coli ATCC pneumoniae No. coli ATCC pneumoniae 25922 ATCC43816 25922 ATCC43816 F-1 >24 >24 020 0.14 0.14 001A 1.0 1.0 021 0.36 0.36 001B 2.0 2.0 022 0.72 1.5 002 0.91 1.4 023 0.30 0.30 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 1.9 3.8 024 0.18 0.21A 0.69 0.52 025 0.16 0.16 B 1.4 1.4 026 0.085 0.085 A 1.0 2.0 027 0.20 0.20 B 0.51 0.51 028 0.72 0.72 0.89 0.89 029 0.12 0.95 1.8 1.8 030 0.30 0.15 1.7 1.8 031 0.16 0.082 1.8 1.8 032 0.16 0.32 0.33 0.33 033 0.56 0.56 2.8 1.4 034 0.29 0.58 035 0.17 0.34 0.91 0.91 036 0.34 0.17 0.46 0.91 037 1.0 2.0 0.54 0.54 038 0.34 0.34 1.4 1.5 039 0.28 0.42 0.56 0.56 040 0.50 2.0 0.28 0.28 041 2.3 4.6 0.55 0.55 042 0.29 0.44 F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Example A Film coated tablets containing the following ingredients can be manufactured in a conventional manner: Ingredients Per tablet Kernel: Compound of formula (I) or a 10.0 mg 200.0 mg pharmaceutically acceptable salt thereof Microcrystalline cellulose 23.5 mg 43.5 mg Lactose hydrous 60.0 mg 70.0 mg Povidone K30 12.5 mg 15.0 mg Sodium starch glycolate 12.5 mg 17.0 mg Magnesium stearate 1.5 mg 4.5 mg (Kernel Weight) 120.0 mg 350.0 mg Film Coat: Hydroxypropyl methyl cellulose 3.5 mg 7.0 mg Polyethylene glycol 6000 0.8 mg 1.6 mg Talc 1.3 mg 2.6 mg Iron oxide (yellow) 0.8 mg 1.6 mg Titan dioxide 0.8 mg 1.6 mg The active ingredient is sieved and mixed with microcrystalline cellulose and the mixture is granulated with a solution of polyvinylpyrrolidone in water. The granulate is then mixed with F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 sodium starch glycolate and magnesium stearate and compressed to yield kernels of 120 or 350 mg respectively. The kernels are lacquered with an aqueous solution / suspension of the above mentioned film coat. Example B Capsules containing the following ingredients can be manufactured in a conventional manner: Ingredients Per capsule Compound of formula (I) or a 25.0 mg pharmaceutically acceptable salt thereof Lactose 150.0 mg Maize starch 20.0 mg Talc 5.0 mg The components are sieved and mixed and filled into capsules of size 2. Example C Injection solutions can have the following composition: Compound of formula (I) or a 3.0 mg pharmaceutically acceptable salt thereof Polyethylene glycol 400 150.0 mg Acetic acid q.s. ad pH 5.0 Water for injection solutions ad 1.0 ml F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 The active ingredient is dissolved in a mixture of Polyethylene glycol 400 and water for injection (part). The pH is adjusted to 5.0 by addition of acetic acid. The volume is adjusted to 1.0 ml by addition of the residual amount of water. The solution is filtered, filled into vials using an appropriate overage and sterilized.

Claims

F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Claims 1. A compound of formula (I) wherein L is absent or alkyl; X is -O- or -NH-; n is 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 heterocyloalkyl, wherein heterocycloalkyl and cycloalkyl are optionally substituted by one or two substituents independently selected from R6; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 with the proviso that if R1 is amino, then L is absent; R2 and R3 are independently selected from halogen; or R2 and R3 together with the carbon they are attached to form cycloalkyl; R4 is halogen or alkyl; R5 is amino, hydroxyalkyl, aminoalkyl, alkylamino, aminoalkylamino or piperazinyl; R6 is at each instance independently selected from amino, alkyl, hydroxyl, aminoalkyl, aminocarbonylalkyl and aminoalkylcarbonylamino; R7 is hydrogen or hydroxy; A1 is -N- or -CH-; and A₂ is -O- or -CH₂-; or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1, wherein L is absent. 3. A compound according to claim 1 or 2, wherein L is alkyl, in particular methyl. 4. A compound according to any one of claims 1 to 3, wherein n is 1. 5. A 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. A compound according to any one of claims 1 to 6, wherein 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)aminoethylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumpropylamino, F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 (hydroxy)difluoropropylamino, (hydroxy)trifluoropropylamino, (aminopropyl)(aminopropyl)(hydroxycarbonylmethyl)ammoniumethoxy, cyclobutyl, azetidin-3-yl, azetidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl or 5-azoniaspiro[2.4]heptane, wherein cyclobutyl, azetidin-3-yl, azetidin-1-yl, morpholin-4-yl, pyrrolidin-1-yl or 5- azoniaspiro[2.4]heptane are optionally substituted by one or two substituents independently selected from R6; with the proviso that if R1 is amino, then L is absent. 8. A compound according to any one of claims 1 to 6, wherein R1 is heterocycloalkyl optionally substituted by one or two substituents independently selected from R6; 9. A compound according to any one of claims 1 to 8, wherein R2 and R3 are both fluoride; or R2 and R3 together with the carbon they are attached to form cyclopropyl. 10. A compound according to any one of claims 1 to 9, wherein R4 is chloride or methyl. 11. A compound according to any one of claims 1 to 10, wherein R5 is amino or hydroxyalkyl, in particular amino. 12. A compound according to any one of claims 1 to 11, wherein R6 is at each instance independently selected from amino, methyl, hydroxy, aminomethyl, aminocarbonylmethyl and aminoethylcarbonylamino. 13. A compound according to any one of claims 1 to 12, wherein R7 is hydrogen. 14. A compound according to any one of claims 1 to 13, wherein A1 is -CH-. 15. A compound according to any one of claims 1 to 14, wherein A₂ -CH₂-. 16. A compound selected from Cis-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Trans-(4R)-4-amino-1-[4-[4-[6-chloro-4-[difluoro-[4-(3-hydroxyazetidin-1- yl)cyclohexyl]methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; 2-amino-N-[1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]azetidin-3-yl]propanamide; (4R)-4-amino-1-[4-[4-[4-[[4-(3-aminopropylamino)cyclohexyl]-difluoro-methyl]-6-chloro- 2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide; cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]propanamide; Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide ; trans-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]propanamide; Trans-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide; Cis-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]cyclobutanecarboxamide; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 Cis-2-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3- hydroxy-propanamide; Cis-1-(2-aminoethyl)-3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]urea; Trans-(6S)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin- 1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5,5-dimethyl-5- azoniaspiro[2.4]heptane-6-carboxamide; Trans-(5R,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5- methyl-5-azoniaspiro[2.4]heptane-6-carboxamide; Trans- (5S,6S)-5-(2-amino-2-oxo-ethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-5- methyl-5-azoniaspiro[2.4]heptane-6-carboxamide; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; Rac-(4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; 2-amino-N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1- yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]acetamide; 4-(((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)amino)-N,N-bis(3-aminopropyl)-N- (carboxymethyl)-4-oxobutan-1-aminium; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2-(2-aminoethoxy)acetamide; 3-amino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-2- hydroxypropanamide; 2,3-diamino-N-((1r, 4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1- yl)phenyl)sulfonyl)piperazin-1-yl)-6-chloropyridin-4- yl)difluoromethyl)cyclohexyl)propanamide; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(diaminomethylene)urea; (2-amino-2-oxo-ethyl)-[4-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-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-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoylamino]propyl-bis(3-aminopropyl)- (carboxymethyl)ammonium; 3-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-1,1-bis(3-aminopropyl)urea; 1-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-3-(3-aminopropyl)guanidine; N-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]methyl]-2,2,2-trifluoro-acetamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 2-[bis(3-aminopropyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; 2-[[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]carbamoyloxy]ethyl-bis(3-aminopropyl)- (carboxymethyl)ammonium; (2S,6S)-2,6-bis(aminomethyl)-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]morpholine-4-carboxamide; N-[4-[1-[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]cyclopropyl]cyclohexyl]-4-[bis(2-aminoethyl)amino]butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]-N-hydroxy- butanamide; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4-[bis(3-aminopropyl)amino]butanamide; (4R)-4-amino-1-[4-[4-[6-chloro-4-[[4-[(2,2-difluoro-3-hydroxy-propyl)amino]cyclohexyl]- difluoro-methyl]-2-pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; (3R,4S)-3-amino-N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro-methyl]cyclohexyl]-4- hydroxy-pyrrolidine-1-carboxamide; 1-((1r,4r)-4-((2-(4-((4-((R)-4-amino-2-oxopyrrolidin-1-yl)phenyl)sulfonyl)piperazin-1-yl)- 6-chloropyridin-4-yl)difluoromethyl)cyclohexyl)-3-(2-(bis(2-aminoethyl)amino)ethyl)urea; F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[(4-aminocyclohexyl)-difluoro-methyl]-6-methyl-pyrimidin-2- yl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1-yl]phenyl]sulfonylpiperazin-1-yl]-6- methyl-pyrimidin-4-yl]-difluoro-methyl]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-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)pyrrolidin-2-one; 2-[bis(2-aminoethyl)amino]ethyl N-[4-[[2-[4-[4-[(4R)-4-amino-2-oxo-pyrrolidin-1- yl]phenyl]sulfonylpiperazin-1-yl]-6-chloro-4-pyridyl]-difluoro- methyl]cyclohexyl]carbamate; (4R)-4-amino-1-[4-[4-[4-[[4-(azetidin-1-yl)cyclohexyl]-difluoro-methyl]-6-chloro-2- pyridyl]piperazin-1-yl]sulfonylphenyl]pyrrolidin-2-one; tert-butyl ((3R)-1-(4-((4-(4-(difluoro((1r,4r)-4-((3,3,3-trifluoro-2- hydroxypropyl)amino)cyclohexyl)methyl)-6-methylpyrimidin-2-yl)piperazin-1- yl)sulfonyl)phenyl)-5-oxopyrrolidin-3-yl)carbamate; or a pharmaceutically acceptable salt thereof. 17. A compound according to any one of claims 1 to 16 for use as therapeutically active substance. 18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 and a therapeutically inert carrier. F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland Case: P38193 19. The use of a compound according to any one of claims 1 to 16 for the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. 20. The use of a compound according to any one of claims 1 to 16 for the preparation of a medicament for the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. 21. A compound according to any one of claims 1 to 16 for use in the treatment or prophylaxis of bacterial infection, in particular wherein the bacteria is gram-negative bacteria. 22. A method for the treatment or prophylaxis of bacterial infection, particularly an infection of a gram-negative bacteria, which method comprises administering an effective amount of a compound as defined in 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 is selected from Enterobacteriaceae, Neisseria gonorrhoeae, Haemophilus influenzae, Helicobacter pylorus, Acinetobacter baumannii and Pseudomonas aeruginosa. 24. The use, method or compound for use according to claim 23 wherein the gram-negative bacteria is Enterobacteriaceae, wherein Enterobacteriaceae is Klebsiella pneumoniae or Escherichia coli. 25. The invention as hereinbefore described.