Antimicrobial compounds and methods

Abstract

The present invention relates to compounds having antibacterial activity. The compounds are active against Gram-positive bacteria and Gram-negative bacteria and can be used to treat infections caused by Gram-positive bacteria and Gram-negative bacteria. A process for preparing the compounds is also disclosed.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Application Serial No. 62 / 934,853, filed on November 13, 2019, which is incorporated herein by reference in its entirety.

[0003] Statement of Government Interest

[0004] This invention was made with government support under Grant 1R01AI132304-01 awarded by the National Institutes of Health (NIH). The government has certain rights in this invention. Technical Field

[0005] The present disclosure relates to compounds having antibacterial activity. The present disclosure also relates to methods of treating bacterial infections using the compounds of the present invention. Background of the Invention

[0007] Antimicrobial resistance is a worldwide problem. Both Gram-positive and Gram-negative bacteria are becoming increasingly resistant to antibiotics.

[0008] Gram-positive bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) are resistant to most antibiotics related to penicillin. MRSA strains are often implicated in infections acquired in healthcare facilities and can cause infections in the larger community.

[0009] Gram-negative bacteria are considered more resistant to antibiotics than Gram-positive bacteria because their cell walls are impermeable. According to the National Institutes of Health (NIH), Gram-negative bacteria can cause many types of infections and are transmitted to humans in a variety of ways. Several species, including Escherichia coli, are common causes of foodborne illness. Vibrio cholerae, the bacterium that causes cholera, is a waterborne pathogen. Gram-negative bacteria can also cause respiratory infections, such as certain types of pneumonia, and sexually transmitted diseases, including gonorrhea. Yersinia pestis is the Gram-negative bacterium that causes plague and is transmitted to humans through the bite of infected insects or handling infected animals. See www.niaid.nih.gov / research / gram-negative-bacteria (last accessed November 6, 2020).

[0010] Some types of Gram-negative bacteria are increasingly resistant to existing antibiotic drugs. Some strains are now resistant to many, most or all available treatments, resulting in increased disease and death caused by bacterial infections, and causing rising health care costs. Examples of Gram-negative bacteria that have demonstrated resistance include: Escherichia coli (E.coli), which causes most urinary tract infections; Acinetobacter baumanii, which mainly causes disease in health care settings; Pseudomonas aeruginosa, which causes bloodstream infections and pneumonia in hospitalized patients, and is a common cause of pneumonia in patients with cystic fibrosis; Klebsiella pneumoniae, which causes multiple types of health care-related infections, including pneumonia, urinary tract infections and bloodstream infections; and Neisseria gonorrhoeae, which causes the sexually transmitted disease gonorrhea and is the second most commonly reported infectious disease in the U.S.

[0011] Therefore, new drugs are needed to combat Gram-positive and Gram-negative bacterial infections. Summary of the invention

[0012] These and other needs are met by the present invention, which in one aspect provides a compound of formula I:

[0013]

[0014]

[0015] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0016] Ring A is a 3-8 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2 and oxo;

[0017] J is C1-C6 alkylene or C3-C8 cycloalkylene, any of which is optionally substituted with halo, OH or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently replaced with O, S, SO, SO2 or C=O;

[0018] R x , Ry , R x’ and R y’ Each is independently H, C1-C6 alkyl or amino protecting group;

[0019] Y is a bond, or C1-C6 alkylene optionally substituted with OH, NH2, CN, halo, or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently substituted with O, NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl), N-(C1-C6 alkyl), 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N-(C 1-6 alkyl)(C=O) or (C=O);

[0020] Ring B is a 3-8 membered monocyclic cycloalkylene, a 3-8 membered monocyclic heterocycloalkylene, a 6-12 membered bicyclic cycloalkylene, or a 6-12 membered bicyclic heterocycloalkylene, each of which is optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl;

[0021] L is a bond or C1-C6 alkylene, wherein up to two methylene units of the C1-C6 alkylene can be independently replaced by O, NH, (C=O), NH(C=O), N-(C 1-6 alkyl)(C=O), (C=NH), NH(C=N) or N-(C 1-6 Alkyl) substitution;

[0022] Ring C and the benzene ring to which it is fused together form an 8-12-membered bicyclic arylene group or an 8-12-membered bicyclic heteroarylene group, wherein the bicyclic heteroarylene group has 1-3 heteroatoms independently selected from N, O or S;

[0023] R 1 , R 2 and R 3 Each is independently selected from the group consisting of C1-C6 alkyl, halo, CN, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COO(C1-C6 alkyl), CONH2, C1-C6 haloalkyl, oxo and C1-C6 alkoxy; and

[0024] m, n and p are each independently 0, 1, 2 or 3.

[0025] In another aspect, the present invention provides methods of using a compound of formula I or a pharmaceutically acceptable salt thereof to treat a bacterial infection.

[0026] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0027] In another aspect, the present invention provides a process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof, as depicted in the synthetic schemes. DETAILED DESCRIPTION

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those of ordinary skill in the art to which the present invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In addition, the materials, methods and embodiments are only illustrative and not intended to be restrictive. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, the present specification including these definitions shall prevail.

[0029] As used herein, the terms "a," "an," and "the" include not only aspects having one member, but also aspects having more than one member.

[0030] As used herein, the term "about" means "approximately" and is used to modify a numerical value, indicating a limited range around the value. If "X" is a value, "about X" will generally indicate a value from 0.95X to 1.05X. Any reference to "about X" specifically indicates at least values ​​X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Therefore, "about X" is intended to teach and provide written description support for claim limitations such as "0.98X". When the quantity "X" only includes integer values ​​(such as "X carbons"), "about X" indicates from (X-1) to (X+1). In this case, "about X" as used herein specifically indicates at least values ​​X, X-1, and X+1.

[0031] When "about" is applied to the beginning of a numerical range, it applies to both ends of the range. Thus, "about 5% to 20%" is equivalent to "about 5% to about 20%". When "about" is applied to the first value of a group of values, it applies to all values ​​in that group. Thus, "about 7%, 9% or 11%" is equivalent to "about 7%, about 9% or about 11%".

[0032] As used herein, a wavy line drawn on a structure may be used to show the connection points of the structure, e.g. in Indicates the connection point.

[0033] The term "acyl" as used herein includes alkanoyl, aroyl, heterocyclic acyl or heteroaroyl as defined herein. Examples of acyl include, but are not limited to, acetyl, benzoyl and nicotinoyl.

[0034] The term "alkanoyl" as used herein includes an alkyl-C(O)- group, wherein the alkyl group is as defined herein. Examples of alkanoyl groups include, but are not limited to, acetyl and propionyl.

[0035] As used herein, the term "agent" includes a compound or mixture of compounds that, when added to a composition, tends to have a particular effect on the properties of the composition. For example, a composition comprising a thickening agent may be more viscous than an otherwise identical comparative composition lacking the thickening agent.

[0036] As used herein, the term "alkyl" includes aliphatic hydrocarbon chains that may be straight or branched. The chain may contain the indicated number of carbon atoms: for example, C1-C 10 Indicates that the group may have 1 to 10 (inclusive) carbon atoms therein. If not otherwise indicated, the alkyl contains 1 to about 20 carbon atoms. In some aspects, the alkyl has 1 to about 10 carbon atoms. In some aspects, the alkyl ("low alkyl") has 1 to 8, 1 to 6, or 1 to 3 carbon atoms in the chain. Examples may include, but are not limited to, methyl, ethyl, propyl, isopropyl (iPr), 1-butyl, 2-butyl, isobutyl (iBu), tert-butyl, pentyl, 2-methylbutyl, 1,1-dimethylpropyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, cyclopentyl, or cyclohexyl.

[0037] Alkyl can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms (e.g., 1 to 4, 1 to 2 or 1) of the alkyl can be independently selected from the group consisting of: fluorine, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio. In some respects, the alkyl is unsubstituted or not optionally substituted.

[0038] As used herein, "alkylene" includes alkyl groups substituted at two points. An example is methylene (-CH2-), propylene (-CH2CH2CH2-), etc.

[0039] As used herein, the term "alkenyl" includes straight or branched chain hydrocarbons containing at least one carbon-carbon double bond. The chain may contain the indicated number of carbon atoms. For example, "C1-C 12"Alkenyl" indicates that the group may have 1 to 12 (inclusive) carbon atoms and at least one carbon-carbon double bond. When the number of carbon atoms indicated is 1, then C i Alkenyl is double bonded to carbon (i.e. carbon is equivalent to oxo). In some aspects, the chain comprises 1 to 12, about 2 to 15, about 2 to 12, about 2 to 8 or about 2 to 6 carbon atoms. Alkenyl can preferably be a stereoisomer (i.e., cis- or alternatively, trans-). The example of alkenyl can include but is not limited to vinyl (ethenyl) (i.e. vinyl (vinyl)), allyl, propenyl, butenyl, crotyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, dodecenyl, cyclopentenyl, cyclohexenyl, 2-isopentenyl, propadienyl, butadienyl, pentadienyl, 3- (1,4-pentadienyl) and hexadienyl.

[0040] Alkenyl can be unsubstituted or optionally substituted.When optionally substituted, one or more hydrogen atoms (for example, 1 to 4, 1 to 2 or 1) of alkenyl can be independently selected from the part of the group consisting of: fluorine, hydroxyl, alkoxyl, amino, alkylamino, acylamino, sulfo and alkylthio, under the condition that there is no hydrogen atom substituent on the carbon-carbon double bond to be replaced by hydroxyl, amino or sulfo group.In some respects, alkenyl is unsubstituted or is not optionally substituted.

[0041] As used herein, "alkenylene" includes alkenyl groups substituted at two points. An example is but-2-enyl (-CH2CH=CHCH2-) and the like.

[0042] As used herein, the term "alkoxy" includes a straight or branched chain saturated or unsaturated hydrocarbon containing at least one oxygen atom in the ether group (e.g., EtO-). The chain may contain the indicated number of carbon atoms. For example, "C1-C 12 "Alkoxy" indicates that the group may have 1 to 12 (inclusive) carbon atoms and at least one oxygen atom. 12 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, butoxy, n-pentoxy, isopentoxy, neopentoxy, and hexoxy.

[0043] Alkoxy can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms (e.g., 1 to 4, 1 to 2 or 1) of alkoxy can be independently selected from the group consisting of: fluorine, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio, provided that the α-position hydrogen atom without ether oxygen is replaced by hydroxyl, amino or thio groups. In some respects, alkoxy is unsubstituted or not optionally substituted.

[0044] As used herein, the term "alkynyl" includes straight, branched or cyclic hydrocarbons containing at least one carbon-carbon triple bond. Examples may include, but are not limited to, ethynyl, propargyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl or decynyl.

[0045] As used herein, "alkynylene" includes alkynyl groups substituted at two points. An example is 2-butynylene (-CH2CCCH2-) and the like.

[0046] Alkynyl can be unsubstituted or optionally substituted. When optionally substituted, one or more hydrogen atoms (e.g., 1 to 4, 1 to 2 or 1) of the alkynyl can be independently selected from the group consisting of: fluorine, hydroxyl, alkoxyl, amino, alkylamino, acylamino, thio and alkylthio, provided that no sp-hybridized hydrogen atom substituent is replaced by hydroxyl, amino or thio groups. In some aspects, the alkynyl is unsubstituted or not optionally substituted.

[0047] As used herein, "aryl" refers to monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthyl, tetrahydronaphthyl, dihydroindenyl); and tricyclic (e.g., fluorenyl, tetrahydrofluorenyl, tetrahydroanthracenyl or anthracenyl) ring systems, wherein the monocyclic ring system is aromatic, or at least one ring in the bicyclic or tricyclic ring system is aromatic. The other rings in the bicyclic or tricyclic ring system can be saturated, partially unsaturated or fully unsaturated. Bicyclic and tricyclic groups include benzo-fused carbocyclic rings. For example, benzo-fused groups include a benzene ring fused to one or two 4- to 8-membered carbocyclic ring portions (e.g., 1,2,3,4-tetrahydronaphthalene or 2,3-dihydro-1H-indene ring systems).

[0048] The term "arylene" as used herein includes aryl groups substituted at two points.

[0049] Aryl can be unsubstituted or optionally substituted.When optionally substituted, one or more hydrogen atoms (for example, 1 to 5, 1 to 2 or 1) of aryl can be independently selected from the part of the group consisting of: alkyl, cyano, acyl, halide, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio.In some respects, alkoxy is unsubstituted or not optionally substituted.

[0050] The term "arylalkyl" or "aralkyl" as used herein includes alkyl groups as defined herein in which at least one hydrogen substituent has been replaced by an aryl group as defined herein. Examples include, but are not limited to, benzyl, 1-phenylethyl, 4-methylbenzyl, and 1,1,-dimethyl-1-phenylmethyl.

[0051] Depending on its component groups, arylalkyl or aralkyl can be unsubstituted or optionally substituted. For example, but not limited to, the aryl group of arylalkyl can be substituted, such as in 4-methylbenzyl. In some aspects, the group is unsubstituted or not optionally substituted, especially in the case of including defined substituents, such as hydroxyalkyl or alkylaminoalkoxy.

[0052] As used herein, the term "cycloalkyl" includes non-aromatic saturated monocyclic or polycyclic ring systems, which may contain a specified number of carbon atoms, wherein none of the rings in the polycyclic ring system is aromatic. 12 Indicates that the group may have 3 to 12 (inclusive) carbon atoms therein. If not otherwise indicated, the cycloalkyl contains about 3 to about 20 carbon atoms. In some aspects, the cycloalkyl has 3 to about 12 carbon atoms in the group. In some aspects, the cycloalkyl has 3 to about 7 carbon atoms in the group. Examples may include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 4,4-dimethylcyclohexyl, and cycloheptyl. The term "cycloalkyl" also includes polycyclics, such as bicyclic cycloalkyls or tricyclic cycloalkyls, which may be fused, bridged, or spiro oriented.

[0053] Cycloalkyl can be unsubstituted or optionally substituted.When optionally substituted, one or more hydrogen atoms (e.g., 1 to 4, 1 to 2 or 1) of cycloalkyl can be independently selected from the group consisting of: fluorine, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio.In some respects, the substituted cycloalkyl can be incorporated into an outer ring or an inner ring olefin (e.g., cyclohex-2-ene-1-yl).In some respects, cycloalkyl is unsubstituted or is not optionally substituted.

[0054] The term "cycloalkylene" as used herein includes cycloalkyl groups substituted at two points.

[0055] The terms "disorder" and "disease" are used interchangeably herein to refer to a condition of a subject. A disorder is a disturbance or disorder that affects the normal function of a subject's body. A disease is a pathological condition of an organ, body part, or system resulting from a variety of causes, such as infection, genetic defects, or environmental stress, characterized by an identifiable set of symptoms. A disorder or disease may refer to a biofilm-related disorder or a disorder caused by a planktonic bacterial phenotype characterized by disease-associated bacterial growth.

[0056] The term "effective amount" or "effective dose" as used herein includes an amount sufficient to achieve the desired result and will therefore depend on the ingredients and their desired results. Nevertheless, once the desired effect is identified, determining the effective amount is within the skill of those skilled in the art.

[0057] As used herein, "fluoroalkyl" includes alkyl groups wherein the alkyl group contains one or more fluorine substituents. Examples include, but are not limited to, trifluoromethyl.

[0058] As used herein, "geminal" substitution includes two or more substituents directly attached to the same atom. An example is 3,3-dimethyl substitution on a cyclohexyl or spirocyclohexyl ring.

[0059] As used herein, "halo" or "halogen" includes fluorine, chlorine, bromine and iodine.

[0060] As used herein, "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic ring system having 4 to 15 ring atoms, wherein one or more of the ring atoms is a heteroatom (e.g., N, O, S, or a combination thereof), and wherein the monocyclic ring system is aromatic, or at least one of the rings in the bicyclic or tricyclic ring system is aromatic. The other rings in the bicyclic or tricyclic ring system may be saturated, partially unsaturated, or fully unsaturated. The heteroaryl group may comprise a benzo-fused ring system having 2 to 3 rings. For example, a benzo-fused group comprises a benzo fused to one or two 4 to 8 membered heterocyclic moieties. Some examples of heteroaryl groups are azetidinyl, pyridinyl, 1H-indazolyl, furanyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuranyl, isoquinolyl, benzothiazolyl, xanthene, thioxanthene, phenothiazine, indoline, benzo[1,3]dioxole, benzo[b]furanyl, benzo[b]phenylthio, indazolyl, benzimidazolyl, benzothiazolyl, purine In some embodiments, the heteroaryl group may include chromanyl, isoindolyl, and tetrahydroisoquinolyl.

[0061] The term "heteroarylene" as used herein includes heteroaryl groups that are substituted at two points.

[0062] Heteroaryl can be unsubstituted or optionally substituted.When optionally substituted, one or more hydrogen atoms (e.g., 1 to 5, 1 to 2 or 1) of heteroaryl can be independently selected from the group consisting of: alkyl, cyano, acyl, halide, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio.In some respects, heteroaryl is unsubstituted or is not optionally substituted.

[0063] As used herein, "heterocycloalkyl" includes non-aromatic rings of about 3 to about 15 ring atoms (e.g., 5 to about 10 ring atoms or 3 to about 6 ring atoms), wherein one or more atoms in the ring system is one or more elements other than carbon, such as nitrogen, oxygen or sulfur. The heterocycloalkyl group optionally contains at least one sp 2 -heteroatoms (e.g., rings incorporating carbonyl, endocyclic olefins, or exocyclic olefins). In some embodiments, the nitrogen or sulfur atom of heterocycloalkyl is optionally oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. Monocyclic heterocycle refers to a three-, four-, five-, six-, seven-, or eight-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S. The three- or four-membered ring contains zero or one double bond and a heteroatom selected from the group consisting of O, N, and S. The five-membered ring contains zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. The six-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. The seven- and eight-membered rings contain zero, one, two, or three double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperidinyl, oxazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyridazin-3(2H)-onyl, pyridin-2(1H)-onyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl.

[0064] As used herein, the term "heterocycloalkylene" includes heterocycloalkyl groups substituted at two points.

[0065] The term "heterocycloalkyl" also includes polycyclic ring systems such as bicyclic heterocycles or tricyclic heterocycles, which may be in fused, bridged or spiro orientations, wherein none of the rings in the polycyclic ring system is aromatic. The bicyclic heterocycle may be a monocyclic heterocycle fused to a monocyclic cycloalkyl, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl, or a monocyclic heterocycle fused to a monocyclic heterocycle, or a bridged monocyclic heterocycle ring system, wherein two non-adjacent atoms of the ring are linked by an alkylene bridge of 1, 2, 3 or 4 carbon atoms or an alkenylene bridge of 2, 3 or 4 carbon atoms. Representative examples of bicyclic heterocycles include, but are not limited to, 3-azabicyclo[3.1.0]hexane, 3-azabicyclo[4.1.0]heptane, 3-azabicyclo[3.2.0]heptane, (3aR,6aS)-hexahydro-1H-2λ2-cyclopenta[c]pyrrole, (3aR,7aS)-octahydro-2λ2-isoindole.

[0066] Heterocycloalkyl can be unsubstituted or optionally substituted.When optionally substituted, one or more hydrogen atoms (e.g., 1 to 4, 1 to 2 or 1) of the group can be independently selected from the group consisting of: fluorine, hydroxyl, alkoxy, amino, alkylamino, acylamino, thio and alkylthio.In some respects, the substituted heterocyclic radical can be incorporated into an outer ring or an inner ring olefin (e.g., cyclohex-2-ene-1-yl).In some respects, heterocyclic radical is unsubstituted or is not optionally substituted.

[0067] The monocyclic heterocycle, bicyclic heterocycle, and tricyclic heterocycle are attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the rings, and can be unsubstituted or substituted.

[0068] As used herein, the term "hydrophilic moiety" or "hydrophilic group" includes moieties or functional groups that have a strong affinity for water. Examples may include, but are not limited to, charged moieties, such as cationic or anionic moieties, or polar uncharged moieties, such as alkoxy or amine groups.

[0069] As used herein, the term "hydroxyalkyl" includes an alkyl group in which at least one hydrogen substituent has been replaced by an alcohol (-OH) group. In some aspects, the hydroxyalkyl group has one alcohol group. In some aspects, the hydroxyalkyl group has one or two alcohol groups, each of which is located on a different carbon atom. In some aspects, the hydroxyalkyl group has 1, 2, 3, 4, 5, or 6 alcohol groups. Examples may include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, and 1-hydroxyethyl.

[0070] When any two substituents, or any two instances of the same substituent, are "independently selected" from a list of alternatives, the groups may be the same or different. a and R b are independently selected from the group consisting of: alkyl, fluoro, amino and hydroxyalkyl, then there are two R a Group and two Rb All groups of the molecule of the group may be alkyl groups (e.g., four different alkyl groups). a Can be an alkyl group, the second R a It can be fluorine, the first R b can be a hydroxyalkyl group, and the second R b It may be amino (or any other substituent taken from said group). Alternatively, R a and the first R b Both can be fluorine, and the second R b It may be an alkyl group (ie, some pairs of substituents may be the same, while other pairs may be different).

[0071] An "amino protecting group" is a protecting group suitable for preventing undesired reactions at the amino nitrogen. Representative amino protecting groups include, but are not limited to, formyl; acyl, e.g., alkanoyl, such as acetyl and trifluoroacetyl; alkoxycarbonyl, such as tert-butoxycarbonyl (Boc) and methoxycarbonyl; arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr) and 1,1-bis-(4'-methoxyphenyl)methyl; and the like.

[0072] A "hydroxy protecting group" is a protecting group suitable for preventing undesired reactions at the hydroxy oxygen. Representative hydroxy protecting groups include, but are not limited to, acyl groups, e.g., alkanoyl groups, such as acetyl; arylmethyl groups, such as benzyl (Bn), trityl (Tr), and 1,1-bis-(4'-methoxyphenyl)methyl; silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBDMS or TBS); and the like.

[0073] As used herein, the term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reactions, etc. and commensurate with a reasonable benefit / risk ratio within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which are incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed of amino groups with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or salts formed by using other methods used in the art (such as ion exchange methods). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

[0074] "Pharmaceutically acceptable acid addition salts" refers to those salts which retain the biological effectiveness of the free bases and which are not biologically or otherwise undesirable, and which are formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as organic acids, such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, orotic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0075] "Pharmaceutically acceptable base addition salts" include those derived from inorganic bases, such as sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Exemplary salts are ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include, but are not limited to, 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-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Exemplary organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. (See, e.g., SM Berge et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19, which is incorporated herein by reference.)

[0076] As used herein, "or" should generally be interpreted non-exclusively. For example, an embodiment of "a composition comprising A or B" will generally present aspects of a composition comprising both A and B. However, "or" should be interpreted to exclude the presentation of those aspects that cannot be combined without contradiction (e.g., a composition pH between 9 and 10 or between 7 and 8).

[0077] As used herein, "spiro bicyclic cycloalkyl" includes cycloalkyl groups in which the geminal substituents on the carbon atoms are replaced to form a 1,1-substituted ring. For example, but not limited to, for -C(R 1 )(R 2 )-group, if R 1 and R 2 Connect to form a cyclopropyl ring, the cyclopropyl ring is incorporated into R 1 and R 2 If the carbon to which it is bonded is a spiro bicyclic cycloalkyl group (ie, a spirocyclopropyl group), then this would be a spiro bicyclic cycloalkyl group (ie, a spirocyclopropyl group).

[0078] As used herein, the term "spiro bicyclic cycloalkylene" includes spiro bicyclic cycloalkyl groups substituted at two points.

[0079] As used herein, "spiro bicyclic heterocycloalkyl" includes heterocycloalkyl groups in which the geminal substituents on carbon atoms are replaced to form a 1,1-substituted ring. For example, but not limited to, for -C(R) as part of a longer carbon chain 1 )(R 2 )-group, if R 1 and R2 Connect to form a pyrrolidine ring, the pyrrolidine ring is incorporated into R 1 and R 2 If the carbon to which it is bonded is cycloheterocycloalkyl, then this would be a spiro bicyclic heterocycloalkyl.

[0080] As used herein, the term "spiro bicyclic heterocycloalkylene" includes spiro bicyclic heterocycloalkyl groups substituted at two points.

[0081] Some of the compounds disclosed herein are characterized by the presence of an amino functional group. Thus, one of ordinary skill will appreciate that the compounds can be isolated as salts in which the nitrogen of the amino functional group is quaternized.

[0082] As used herein, the terms "treat," "treating," or "treatment" include administering or applying a composition (e.g., a composition described herein) in an amount, manner (e.g., administration schedule), and mode (e.g., administration route) that is effective to improve a condition or a symptom thereof, or to delay or slow the progression of a condition or a symptom thereof. Such improvements may include, but are not limited to, alleviation or improvement of one or more symptoms or disorders, alleviation of the extent of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, reduction in disease recurrence, and remission, whether partial or complete, and whether detectable or undetectable.

[0083] Implementation

[0084] Compound

[0085] In a first aspect, the present disclosure provides a compound of formula I:

[0086]

[0087] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0088] Ring A is a 3-8 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2 and oxo;

[0089] J is C1-C6 alkylene or C3-C8 cycloalkylene, any of which is optionally substituted with halo, OH or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently replaced with O, S, SO, SO2 or C=O;

[0090] R x , R y , R x’ and R y’ Each is independently H, C1-C6 alkyl or amino protecting group;

[0091] Y is a bond or a C1-C6 alkylene optionally substituted with OH, NH2, CN, halo or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently substituted with O, NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl), N-(C1-C6 alkoxy) 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N-(C 1-6 alkyl)(C=O) or (C=O);

[0092] Ring B is a 3-8 membered monocyclic cycloalkylene, a 3-8 membered monocyclic heterocycloalkylene, a 6-12 membered bicyclic cycloalkylene, or a 6-12 membered bicyclic heterocycloalkylene, each of which is optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), COONH2, COONH(C1-C6 alkyl), COON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl;

[0093] L is a bond or C1-C6 alkylene, wherein up to two methylene units of the C1-C6 alkylene can be independently replaced by O, NH, (C=O), NH(C=O), N-(C 1-6 alkyl)(C=O), (C=NH), NH(C=N) or N-(C 1-6 Alkyl) substitution;

[0094] Ring C and the benzene ring to which it is fused together form an 8-12-membered bicyclic arylene group or an 8-12-membered bicyclic heteroarylene group, wherein the bicyclic heteroarylene group has 1-3 heteroatoms independently selected from N, O or S;

[0095] R 1 , R 2 and R 3 Each is independently selected from the group consisting of C1-C6 alkyl, halo, CN, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COO(C1-C6 alkyl), COONH2, C1-C6 haloalkyl, oxo and C1-C6 alkoxy; and

[0096] m, n and p are each independently 0, 1, 2 or 3.

[0097] In one embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring A is a 5-6 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2 and oxo.

[0098] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring A is Each R 4 Independently selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2 and oxo, wherein q is 0, 1 or 2.

[0099] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring A is

[0100] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, J is C1-C6 alkylene optionally substituted with halo, OH or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently replaced by O, S, SO, SO2 or C=O.

[0101] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, J is C1-C6 alkylene optionally substituted with OH, wherein one methylene unit of the C1-C6 alkylene is replaced by C=O.

[0102] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, J is

[0103] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, R x and R y is H. In another embodiment, R x is H, and R y is C1-C6 alkyl. In another embodiment, R x is H, and R yis an amino protecting group. In another embodiment, R x is H, and R y is a Boc group or a methoxycarbonyl group. In another embodiment, R x and R y Each is independently C1-C6 alkyl. In another embodiment, R x and R y Each is independently selected from the group consisting of H, Boc and methoxycarbonyl.

[0104] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, yes

[0105] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, yes

[0106] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring C together with the benzene ring to which it is fused forms a 9-10 membered bicyclic arylene or a 9-10 membered bicyclic heteroarylene, wherein the bicyclic heteroarylene has 1-3 heteroatoms independently selected from N, O and S.

[0107] In another embodiment, ring C together with the benzene ring to which it is fused forms a 9-10 membered bicyclic arylene or a 9-10 membered bicyclic heteroarylene, wherein the bicyclic heteroarylene has 1-3 heteroatoms independently selected from N and O.

[0108] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Selected from the group consisting of: wherein each X1 is independently selected from the group consisting of: CH2, CH, O, S, SO, SO2, N and NH; R 2 and R 3 Each is independently C1-C6 alkyl, halo or C1-C6 haloalkyl; and n and p are each independently 0, 1 or 2. In another embodiment, each X1 is independently selected from the group consisting of CH2, CH, O, N and NH; R 2 and R 3 are each independently C1-C6 alkyl, halo or C1-C6 haloalkyl; and n and p are each independently 0 or 1.

[0109] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, yes wherein X1 is CH2, O, S, SO, SO2 or NH, and wherein R 2 and R 3 Each is independently C1-C6 alkyl, halo or C1-C6 haloalkyl, wherein n and p are each independently 0, 1 or 2.

[0110] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Selected from the group consisting of:

[0111] In another embodiment, yes

[0112] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Selected from the group consisting of:

[0113] In another embodiment, yes

[0114] In another embodiment of the compound of Formula I, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Y is a bond.

[0115] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Y is C1-C3 alkylene optionally substituted with OH, NH2, halo or C1-C6 alkoxy, and wherein one methylene unit of the C1-C3 alkylene is optionally substituted with O, NH, N(C1-C6 alkyl), N(C1-C6 hydroxyalkyl), N(C1-C6 haloalkyl), N(C1-C6 alkoxy) 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N(C 1-6 alkyl)(C═O) or (C═O).

[0116] In another embodiment of the compound of Formula I or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Y is selected from the group consisting of: O, NH, NH-C 1-2 Alkylene, N(C1-C6 alkyl), N(C1-C6 hydroxyalkyl), N(C1-C6 haloalkyl), N(C1-C6 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N(C 1-6 alkyl)(C=O) and (C=O).

[0117] In another embodiment, Y is O, NH, N(C1-C6 alkyl), N(C1-C6 hydroxyalkyl), N(C1-C6 haloalkyl), N(C1-C6 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N(C 1-6 alkyl)(C═O) or (C═O).

[0118] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Y is -NH-, -N(C1-C6 alkyl)- or -N(C 1-6 Alkylene-C 3-8 Cycloalkyl)-.

[0119] In another embodiment of the compound of Formula I or a single stereoisomer or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Y is selected from the group consisting of: -NH-, -N(C1-C3 alkyl)-, -NH-C 1-2 Alkylene- and -N(C 1-3 Alkylene-C 3-6 In another embodiment, Y is selected from the group consisting of —NH—, —NMe—, —NEt—, —NH—CH 2 —, and —N(CH 2 -cyclopropyl)—.

[0120] In another embodiment, Y is selected from the group consisting of: a bond, -NH-, -N(C1-C3 alkyl)-, -NH-C 1-2 Alkylene- and -N(C 1-3 Alkylene-C 3-6 In another embodiment, Y is selected from the group consisting of a bond, -NH-, -NMe-, -NEt-, -NH-CH2-, and -N(CH2-cyclopropyl)-.

[0121] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Selected from the group consisting of:

[0122] In another embodiment, yes

[0123] In another embodiment of the compound of Formula I, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 4-7 membered monocyclic cycloalkylene, a 4-7 membered monocyclic heterocycloalkylene, a 6-10 membered bicyclic cycloalkylene, or a 6-10 membered bicyclic heterocycloalkylene, each of which is optionally substituted with up to three substituents independently selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

[0124] In another embodiment, Ring B is a 4-7 membered monocyclic cycloalkylene, a 4-7 membered monocyclic heterocycloalkylene, a 6-10 membered bicyclic cycloalkylene, or a 6-10 membered bicyclic heterocycloalkylene.

[0125] In another embodiment, Ring B is a 4-6 membered monocyclic cycloalkylene, a 4-7 membered monocyclic heterocycloalkylene, or a 6-7 membered bicyclic cycloalkylene or a 6-7 membered bicyclic heterocycloalkylene.

[0126] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 3-8 membered monocyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

[0127] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 4-6 membered monocyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl.

[0128] In another embodiment, Ring B is a 3-8 membered monocyclic cycloalkylene. In another embodiment, Ring B is a 4-7 membered monocyclic cycloalkylene. In another embodiment, Ring B is selected from the group consisting of cyclobutylene, cyclopentylene, and cyclohexylene.

[0129] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 3-8 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

[0130] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 4-7 membered monocyclic heterocycloalkylene optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl, and Ring B contains up to 2 nitrogen atoms.

[0131] In another embodiment, Ring B is a 3-8 membered monocyclic heterocycloalkylene. In another embodiment, Ring B is a 5-7 membered monocyclic heterocycloalkylene, and Ring B contains 2 nitrogen atoms. In another embodiment, Ring B is a 4-7 membered monocyclic heterocycloalkylene, wherein Ring B contains 1 nitrogen atom. In another embodiment, Ring B is a 5-7 membered monocyclic heterocycloalkylene, wherein Ring B contains 1 nitrogen atom.

[0132] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 6-10 membered bicyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl.

[0133] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a 6-10 membered fused, spiro or bridged bicyclic cycloalkylene.

[0134] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a 6-10 membered fused bicyclic cycloalkylene.

[0135] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a 6-10 membered bridged bicyclic cycloalkylene.

[0136] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a 6-10 membered spiro bicyclocycloalkylene. In another embodiment, Ring B is a 7 membered spiro bicyclocycloalkylene.

[0137] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 6-12 membered bicyclic heterocycloalkylene optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, C1-C6 haloalkyl, OH, and C1-C6 hydroxyalkyl. In another embodiment, Ring B is a 6-12 membered bicyclic heterocycloalkylene.

[0138] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a 6-12 membered fused, spiro or bridged bicyclic heterocycloalkylene containing up to 3 nitrogen atoms.

[0139] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is a 6-12 membered fused bicyclic heterocycloalkylene containing up to 2 nitrogen atoms. In another embodiment, Ring B is a 6-10 membered fused bicyclic heterocycloalkylene containing 2 nitrogen atoms. In another embodiment, Ring B is a 6-10 membered fused bicyclic heterocycloalkylene containing 1 nitrogen atom. In another embodiment, Ring B is a 6-10 membered fused bicyclic heterocycloalkylene containing 1 nitrogen atom. In another embodiment, Ring B is a 6 membered fused bicyclic heterocycloalkylene containing 1 nitrogen atom.

[0140] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a spirobicyclic heterocycloalkylene containing up to 2 nitrogen atoms.

[0141] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, Ring B is a bridged bicyclic heterocycloalkylene containing up to 2 nitrogen atoms.

[0142] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Ring B is selected from the group consisting of:

[0143] In another embodiment, Ring B is selected from the group consisting of:

[0144] In another embodiment of the compound of Formula I, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, L is a bond.

[0145] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, L is C1-C6 alkylene, wherein up to two methylene units of said C1-C6 alkylene are optionally and independently replaced by O, NH, (C=O), NH(C=O), N-(C 1-6 alkyl)(C=O), (C=NH), NH(C=N) or N-(C 1-6 Alkyl) replacement.

[0146] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, L is C1-C6 alkylene.

[0147] In another embodiment, L is -CH2- or -CH2CH2-. In another embodiment, L is -CH2-. In another embodiment, L is -CH2CH2-.

[0148] In another embodiment, L is a bond or C1-C3 alkylene. In another embodiment, L is a bond, -CH2- or -CH2-CH2-.

[0149] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, R x’ and R y’ is H. In another embodiment, R x’ is H, and R y’ is C1-C6 alkyl. In another embodiment, R x’ is H, and R y’ is an amino protecting group. In another embodiment, R x’ and R y’ Each is independently C1-C6 alkyl. In another embodiment, R x’ and R y’ Each is independently selected from the group consisting of H, Boc and methoxycarbonyl. x’ and R y’ Each is independently selected from the group consisting of H and methoxycarbonyl.

[0150] In another embodiment, Y is C1-C3 alkylene, wherein one methylene unit of the C1-C3 alkylene is optionally replaced by -NH-, -N(C1-C6 alkyl)-, or -N(C 1-6 Alkylene-C 3-8 cycloalkyl)-substituted; Ring B is a 4-6 membered monocyclic cycloalkylene, a 4-7 membered monocyclic heterocycloalkylene, a 6-9 membered bicyclic cycloalkylene or a 6-9 membered bicyclic heterocycloalkylene; L is a bond or a C1-C3 alkylene; and R x’ and R y’ Each is independently H or an amino protecting group.

[0151] In another embodiment, Y is selected from the group consisting of: -NH-, -NMe-, -NEt-, -NH-CH2-, and -N(CH2-cyclopropyl)-; and Ring B is selected from the group consisting of: L is a bond, -CH2- or -CH2-CH2-; and R x’ and R y’ Each is independently selected from the group consisting of H, Boc and methoxycarbonyl.

[0152] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, Selected from the group consisting of:

[0153] In another embodiment, Selected from the group consisting of:

[0154] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3 Each is independently selected from the group consisting of a C1-C6 alkyl group, a halogen group, a C1-C6 haloalkyl group, an oxo group, and a C1-C6 alkoxy group, and m, n, and p are each independently 0, 1, or 2.

[0155] In another embodiment of the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3Each is independently C1-C6 alkyl, halo, oxo or C1-C6 haloalkyl, and m, n and p are each independently 0 or 1. In another embodiment, m, n and p are 0.

[0156] In another embodiment, the compound of formula I is a compound of formula IA:

[0157]

[0158] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, ring C, J, L, Y, R 2 , R 3 , R x , R y , R x’ , R y’ , n and p have the definitions provided in the preceding paragraphs.

[0159] In another embodiment, the compound of Formula I or Formula IA is a compound of Formula IA-1:

[0160]

[0161] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, ring C, J, L, Y, R 2 , R 3 , R x , R y , R x’ , R y’ , n and p have the definitions provided in the preceding paragraph; each R 4 is independently selected from the group consisting of H, C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, NH2 and oxo; and q is 0, 1, 2 or 3.

[0162] In another embodiment, the compound of formula I, IA or IA-1 is a compound of formula IA-2:

[0163]

[0164] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, ring C, L, Y, R 2 , R 3 , R x , R y , R x’ , R y’, n and p have the definitions provided in the preceding paragraph; wherein K is C1-C4 alkylene optionally substituted with halo, hydroxy or C1-C6 alkoxy.

[0165] In another embodiment, the compound of formula I, IA, IA-1 or IA-2 is a compound of formula IA-3:

[0166]

[0167] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, ring C, L, Y, R 2 , R 3 , R x , R y , R x’ , R y’ , n and p have the definitions provided in the preceding paragraph; wherein K is a C1-C3 alkylene group optionally substituted with hydroxy.

[0168] In another embodiment, the compound of formula I, IA, IA-1, IA-2 or IA-3 is a compound of formula IA-4a or IA-4b:

[0169]

[0170] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, L, K, Y, R 2 , R 3 , R x’ , R y’ , n and p have the definitions provided in the preceding paragraph; wherein each X1 is independently selected from the group consisting of CH2, CH, O, S, N and NH.

[0171] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a or IA-4b is a compound of Formula IA-5a or IA-5b:

[0172]

[0173] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, L, K, Y, R x’ and R y’ has the definition provided in the preceding paragraph; wherein each X1 is independently selected from the group consisting of CH2, CH, N, NH and O. In another embodiment, each X1 is independently selected from the group consisting of CH2 and O.

[0174] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-4b, IA-5a or IA-5b is a compound of Formula IA-6a or IA-6b:

[0175]

[0176] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B, L, K, Y and X1 have the definitions provided in the preceding paragraphs.

[0177] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-4b, IA-5a, IA-5b, IA-6a or IA-6b is a compound of Formula IA-7a, Formula IA-7b, Formula IA-7c, Formula IA-7d, Formula IA-7e or Formula IA-7f:

[0178]

[0179]

[0180] or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L, K, Y and X1 have the definitions provided in the preceding paragraphs; each X2 is independently CH or N; and each s is independently 1, 2 or 3.

[0181] In one embodiment, K is

[0182] In one embodiment, Y is a bond, -NH-, -NH-(C1-C5 alkylene)-, -N(C1-C6 alkyl)-, or -N(C1-C6 alkylene-C3-C8 cycloalkyl)-.

[0183] In one embodiment, Y is -NH-, -N(C1-C6 alkyl)-, or -N(C1-C6 alkylene-C3-C8 cycloalkyl)-.

[0184] In another embodiment, L is a bond or C1-C6 alkylene. In another embodiment, L is C1-C6 alkylene.

[0185] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7a is a compound of Formula IA-7a-1, IA-7a-2, IA-7a-3, IA-7a-4, IA-7a-5, IA-7a-6, IA-7a-7 or IA-7a-8:

[0186]

[0187]

[0188] or a pharmaceutically acceptable salt thereof, wherein K, X1 and L are as defined herein.

[0189] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7c is a compound of Formula IA-7c-1, Formula IA-7c-2, Formula IA-7c-3, Formula IA-7c-4, Formula IA-7c-5, Formula IA-7c-6, Formula IA-7c-7 or Formula IA-7c-8:

[0190]

[0191]

[0192] or a pharmaceutically acceptable salt thereof, wherein K, X1, Y and L are as defined herein.

[0193] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7c is a compound of Formula IA-7c-9, Formula IA-7c-10, Formula IA-7c-11 or Formula IA-7c-12:

[0194]

[0195]

[0196] or a pharmaceutically acceptable salt thereof, wherein K, X1, Y and L are as defined herein.

[0197] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7c is a compound of Formula IA-7c-13, Formula IA-7c-14, Formula IA-7c-15 or Formula IA-7c-16:

[0198]

[0199] or a pharmaceutically acceptable salt thereof, wherein K, X1, Y and L are as defined herein.

[0200] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7c is a compound of Formula IA-7c-17, Formula IA-7c-18, Formula IA-7c-19, Formula IA-7c-20, Formula IA-7c-21, Formula IA-7c-22, Formula IA-7c-23 or Formula IA-7c-24:

[0201]

[0202] or a pharmaceutically acceptable salt thereof, wherein K, X1, Y and L are as defined herein.

[0203] In another embodiment, the compound of Formula I, IA, IA-1, IA-2, IA-3, IA-4a, IA-5a, IA-6a or IA-7c is a compound of Formula IA-7c-25, Formula IA-7c-26, Formula IA-7c-27 or Formula IA-7c-28:

[0204]

[0205] or a pharmaceutically acceptable salt thereof, wherein K, X1, Y and L are as defined herein.

[0206] paragraph.

[0207] In one embodiment, K is a C1-C3 alkylene optionally substituted with hydroxy; each X1 is independently selected from the group consisting of CH2, CH, NH, N and O; Y is a bond, NH, -NH(C1-C6 alkylene)-, -N(C1-C6 alkyl)- or -N(C1-C6 alkylene-C3-C8 cycloalkyl)-; L is a bond or C1-C3 alkylene.

[0208] In one embodiment, K is each X1 is independently selected from the group consisting of CH2, CH, NH, N, and O; Y is selected from the group consisting of a bond, -NH-, -NMe-, -NEt-, -NH-CH2-, and -N(CH2-cyclopropyl)-; and L is a bond, -CH2-, or -CH2CH2-.

[0209] In one embodiment, K is each X1 is independently selected from the group consisting of: CH2, CH, NH, N, and O; Y is selected from the group consisting of: a bond, -NH-, -NMe-, -NEt-, -NH-CH2-, and -N(CH2-cyclopropyl)-; L is a bond, -CH2-, or -CH2CH2-; and Ring B is selected from the group consisting of:

[0210]

[0211] In another aspect, the present disclosure provides a compound or a pharmaceutically acceptable salt thereof listed in Table 1. In Table 1, the free base structure and salt structure of the compound are depicted.

[0212] Table 1. Compounds of Formula I

[0213]

[0214]

[0215]

[0216]

[0217] In another aspect, the present disclosure provides compounds listed in Table 2 or pharmaceutically acceptable salts thereof. In Table 2, the free base and salt structures of the compounds are depicted.

[0218] Table 2. Compounds of Formula I continued

[0219]

[0220]

[0221]

[0222] In another embodiment, the compound of Formula I or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof is selected from the compounds listed in any one of Table 1 and Table 2.

[0223] In another aspect, the present disclosure provides a compound of formula II:

[0224]

[0225] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, ring C, L, Y, R 1 , R 2 , R 3 , R x’ , R y’ , m, n and p have the definitions provided in the preceding paragraph.

[0226] In one embodiment, the compound of Formula II or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof is selected from the compounds as depicted in Table 3 below.

[0227] Table 3. Compounds of Formula II

[0228]

[0229]

[0230]

[0231] In another aspect, the present disclosure provides a compound of formula III

[0232]

[0233] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, ring C, L, Y, R 1 , R 2 , R 3 , R x’ , R y’ , m, n and p have the definitions provided in the preceding paragraph.

[0234] In one embodiment, the compound of formula III or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof is selected from the compounds as depicted in Table 4 below.

[0235] Table 4. Compounds of Formula III

[0236]

[0237]

[0238]

[0239] Pharmaceutical compositions and administration

[0240] The present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient. In certain embodiments, the compound of the present invention is provided in the pharmaceutical composition in an effective amount. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a preventive effective amount.

[0241] Pharmaceutically acceptable excipients include any and all solvents, diluents or other liquid vehicles, dispersions, suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc., as long as they are suitable for the specific dosage form required. General considerations in the formulation and / or manufacture of pharmaceutical compositions can be found in, for example, Remington's Pharmaceutical Sciences, Sixteenth Edition, EW Martin (Mack Publishing Co., Easton, Pa., 1980) and Remington: The Science and Practice of Pharmacy, 21st Edition (Lippincott Williams & Wilkins, 2005).

[0242] The pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparation methods include the steps of bringing the compound of the present invention ("active ingredient") into association with a carrier and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, forming and / or packaging the product into a desired single or multiple dose unit.

[0243] The pharmaceutical composition can be prepared, packaged and / or sold in bulk as a single unit dose and / or as a plurality of single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject and / or a convenient fraction of such a dose, such as, for example, half or one-third of such a dose.

[0244] The relative amounts of the active ingredient, pharmaceutically acceptable excipient and / or any additional ingredients in the pharmaceutical composition of the present invention will vary depending on the identity, size and / or condition of the subject being treated and further depending on the route by which the composition is administered. For example, the composition may contain between 0.1% and 100% (w / w) of the active ingredient.

[0245] Pharmaceutically acceptable excipients for making the provided pharmaceutical compositions include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Excipients such as cocoa butter and suppository waxes, colorants, coating agents, sweeteners, flavoring agents and fragrances may also be present in the compositions.

[0246] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, dibasic calcium phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and mixtures thereof.

[0247] Exemplary granulating and / or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0248] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and Veegum [magnesium aluminum silicate]), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [Tween 20], polyoxyethylene sorbitan [Tween 60], polyoxyethylene sorbitan monooleate [Tween 80], sorbitan monopalmitate [Span 40], sorbitan monostearate [Span 60], sorbitan tristearate [Span 65], glyceryl monooleate, sorbitan monooleate [Span 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [Myr 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g. Cremophor), polyoxyethylene ethers (e.g. polyoxyethylene lauryl ether [Brij30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic F 68, poloxapol 188, cetyltrimethylammonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, etc. and / or mixtures thereof.

[0249] Exemplary binders include starches (e.g., corn starch and starch pastes), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., gum arabic, sodium alginate, extracts of Irish moss, panwar gum, ghatti gum, esparto husk mucilage, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum), and larch arabinogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohols, and / or mixtures thereof.

[0250] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives.

[0251] Exemplary antioxidants include alpha-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0252] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and its salts and hydrates (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, etc.), citric acid and its salts and hydrates (e.g., citric acid monohydrate), fumaric acid and its salts and hydrates, malic acid and its salts and hydrates, phosphoric acid and its salts and hydrates, and tartaric acid and its salts and hydrates. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0253] Exemplary antifungal preservatives include butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0254] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and phenethyl alcohol.

[0255] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0256] Other preservatives include tocopherol, tocopherol acetate, deferoxamine mesylate, cetrimonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon and Euxyl. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0257] Exemplary buffers include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glucuronate, calcium glucoheptonate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium valerate, valeric acid, dibasic calcium phosphate, phosphoric acid, tricalcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and mixtures thereof.

[0258] Exemplary lubricants include magnesium stearate, calcium stearate, stearic acid, silicon dioxide, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0259] Exemplary natural oils include almond oil, almond oil, avocado oil, babassu oil, bergamot oil, black currant seed oil, borage oil, juniper oil, chamomile oil, mustard oil, caraway oil, palm wax oil, castor oil, cinnamon oil, cocoa butter oil, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, linseed oil, geraniol oil, gourd oil, grapeseed oil, hazelnut oil, hyssop oil, isopropyl myristate oil, jojoba oil, macadamia oil, lavender oil, lavender oil , lemon oil, litsea cubeba oil, macadamia oil, mallow oil, mango seed oil, meadowfoam seed oil, mink oil, nutmeg oil, olive oil, orange oil, orange roughy oil, palm oil, palm kernel oil, peach kernel oil, peanut oil, poppy seed oil, pumpkin seed oil, rapeseed oil, rice bran oil, rosemary oil, safflower oil, sandalwood oil, camellia oil, savoury oil, sea buckthorn oil, sesame oil, shea butter oil, silicone oil, soybean oil, sunflower oil, tea tree oil, thistle oil, tsubaki oil, vetiver oil, walnut oil and wheat germ oil. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil and mixtures thereof.

[0260] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to active ingredients, liquid dosage forms may also include inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (such as cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and aromatics. In certain embodiments for parenteral administration, the conjugates of the invention are mixed with solubilizing agents such as Cremophor, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

[0261] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersing or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can be sterile injectable solutions, suspensions or emulsions in parenterally acceptable nontoxic diluents or solvents, for example, as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution (USP) and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are generally used as solvents or suspension media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injectable preparations. Injectable preparations can be sterilized, for example, by filtering through a bacterial retention filter or by incorporating a sterilizing agent in the form of a sterile solid composition, which can be dissolved or dispersed in sterile water or other sterile injectable media before use.

[0262] Suitable dispersants or wetting agents (such as Tween 80) and suspending agents can be used to prepare sterile injectable compositions, such as sterile injectable aqueous or oily suspensions, according to techniques known in the art. Sterile injectable preparations can also be sterile injectable solutions or suspensions in non-toxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are conventionally used as solvents or suspension media (e.g., synthetic monoglycerides or diglycerides). Fatty acids such as oleic acid and its glyceride derivatives can be used for the preparation of injections, as natural pharmaceutically acceptable oils such as olive oil or castor oil, especially in their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, or carboxymethyl cellulose or similar dispersants. Other commonly used surfactants (such as Tweens or Spans) or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0263] In order to prolong the effect of the drug, it is often necessary to slow down the absorption of the drug injected subcutaneously or intramuscularly. This can be achieved by using a liquid suspension of a crystalline or amorphous material with low water solubility. The absorption rate of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, delayed absorption of a drug form administered parenterally can be achieved by dissolving or suspending the drug in an oil vehicle.

[0264] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugate of the invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredients.

[0265] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one pharmaceutically acceptable inert excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or the following substances: a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; b) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; c) humectants, such as glycerol; d) disintegrators, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; e) dissolution retardants, such as paraffin; f) absorption promoters, such as quaternary ammonium compounds; g) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; h) absorbents, such as kaolin and bentonite; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may contain a buffer.

[0266] Solid compositions of similar types can use excipients such as lactose or milk sugar and high molecular weight polyethylene glycol as fillers in soft and hard filled gelatin capsules. Solid dosage forms of tablets, dragees, capsules, pills and granules with coatings and shells such as enteric coatings and other coatings known in the field of pharmaceutical formulation can be prepared. They can optionally contain opacifiers and can have a composition in which they release the active ingredient only or preferentially in a certain part of the intestinal tract, optionally in a delayed manner. Examples of usable embedded compositions include polymeric substances and waxes. Solid compositions of similar types can use excipients such as lactose or milk sugar and high molecular weight polyethylene glycol as fillers in soft and hard filled gelatin capsules.

[0267] The active ingredient can be a microencapsulated form with one or more excipients as described above. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared to have coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical formulation. In such solid dosage forms, the active ingredient can be blended with at least one inert diluent such as sucrose, lactose or starch. According to general practice, such dosage forms can include other substances except inert diluents, for example, tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form can include a buffer. They can optionally include an opacifier and can have a composition that they only or preferentially release the active ingredient in a delayed manner in a certain part of the intestinal tract. Examples of usable embedded compositions include polymeric substances and waxes.

[0268] The topical and / or transdermal dosage forms of the compounds of this invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and / or patches. In general, active ingredients are mixed with pharmaceutically acceptable carriers and / or any required preservatives and / or buffers that may be required under aseptic conditions. In addition, the present invention encompasses the use of transdermal patches, which often have the additional advantage of providing controlled delivery of active ingredients to the body. Such dosage forms can be prepared, for example, by dissolving and / or being dispersed in a suitable medium. Alternatively or additionally, rate control can be provided by providing a rate-controlled membrane and / or by dispersing active ingredients in a polymer matrix and / or a gel.

[0269] Suitable devices for delivering the intradermal pharmaceutical compositions described herein include short needle devices, such as those described in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositions can be administered by devices that limit the effective penetration length of the needle into the skin, such as those described in PCT Publication WO 99 / 34850 and their functional equivalents. Jet injection devices that deliver liquid vaccines to the dermis by means of a liquid jet injector and / or by means of a needle that punctures the stratum corneum and produces a jet that reaches the dermis are suitable. Jet injection devices are described, for example, in U.S. Patent Nos. 5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189; 5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335; 5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880; 4,940,460; and in PCT Publications WO 97 / 37705 and WO 97 / 13537. Ballistic powder / particle delivery devices that use compressed gas to accelerate the vaccine in powder form through the outer layers of the skin to the dermis are suitable. Alternatively or additionally, conventional syringes can be used in the classic mantoux method of intradermal administration.

[0270] The pharmaceutical composition of the present invention can be prepared, packaged and / or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may include dry particles comprising an active ingredient and having a diameter in the range of about 0.5 to about 7 nanometers or about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders, which are administered using a device including a dry powder reservoir to which a propellant can be directed to flow to disperse the powder and / or using a self-propelling solvent / powder dispersion container such as a device including an active ingredient dissolved and / or suspended in a low-boiling propellant in a sealed container. Such powders contain particles, wherein at least 98% by weight of the particles have a diameter greater than 0.5 nanometers and at least 95% of the number of particles have a diameter less than 7 nanometers. Alternatively, at least 95% by weight of the particles have a diameter greater than 1 nanometer and at least 90% of the number of particles have a diameter less than 6 nanometers. The dry powder composition may include a solid fine powder diluent such as sugar and is conveniently provided in a unit dosage form.

[0271] Low boiling point propellants typically include liquid propellants having a boiling point below 65° F. at atmospheric pressure. Typically, the propellant may comprise 50% to 99.9% (w / w) of the composition, and the active ingredient may comprise 0.1% to 20% (w / w) of the composition. The propellant may also contain additional ingredients, such as liquid nonionic and / or solid anionic surfactants and / or solid diluents (which may have a particle size of the same order of magnitude as the particles comprising the active ingredient).

[0272] The pharmaceutical composition of the present invention formulated for pulmonary delivery can provide an active ingredient in the form of droplets of solution and / or suspension. Such preparations can be prepared, packaged and / or sold in the form of aqueous and / or dilute alcohol solutions and / or suspensions containing the active ingredient, optionally aseptic, and can be conveniently applied using any spray and / or atomizing device. Such preparations can also include one or more additional ingredients, including but not limited to flavoring agents such as saccharin sodium, volatile oils, buffers, surfactants and / or preservatives such as methyl hydroxybenzoate. The droplets provided by this route of administration can have an average diameter in the range of about 0.1 to about 200 nanometers.

[0273] The formulations described herein as suitable for pulmonary delivery are suitable for intranasal delivery of the pharmaceutical compositions of the present invention. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle size of about 0.2 to 500 microns. This formulation is administered by rapid inhalation through the nasal passage from a powder container held close to the nostrils.

[0274] Preparations for nasal administration may, for example, contain as little as about 0.1% (w / w) and as much as 100% (w / w) of active ingredients, and may contain one or more additional ingredients described herein. The pharmaceutical composition of the present invention can be prepared, packaged or sold in the form of a formulation for buccal administration. Such preparations may, for example, be in the form of tablets and / or lozenges prepared using conventional methods, and may, for example, contain 0.1% to 20% (w / w) active ingredients, with the remainder containing an orally soluble and / or degradable composition and optionally one or more additional ingredients described herein. Alternatively, preparations for buccal administration may contain a powder and / or aerosolized and / or atomized solution and / or suspension containing the active ingredient. Such powdered, aerosolized and / or aerosolized preparations may have an average particle and / or droplet size in the range of about 0.1 to about 200 nanometers when dispersed, and may also contain one or more additional ingredients described herein.

[0275] Although the descriptions of pharmaceutical compositions provided herein are primarily directed to pharmaceutical compositions suitable for administration to humans, the skilled artisan will appreciate that such compositions are generally suitable for administration to animals of all kinds. It is well known to modify pharmaceutical compositions suitable for administration to humans so that the compositions are suitable for administration to various animals, and the ordinary veterinary pharmacologist can design and / or perform such modifications by ordinary experimentation.

[0276] For ease of administration and uniformity of dosage, the compounds provided herein are generally formulated in dosage unit form. However, it will be understood that the total daily dosage of the compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment. The specific therapeutically effective dosage level for any particular subject or organism will depend on various factors, including the disease, disorder and illness being treated and the severity of the disorder; the activity of the specific active ingredient being used; the specific composition being used; the age, weight, general health, sex and diet of the subject; the time of administration, route of administration and the excretion rate of the specific active ingredient being used; the duration of treatment; drugs used in combination or concomitantly with the specific active ingredient being used; and similar factors well known in the medical field.

[0277] In order to implement the method of the present invention, the above-mentioned compound or its pharmaceutical composition can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, rectally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. Generally, the most appropriate route of administration will depend on various factors, including the nature of the agent (e.g., the stability of the agent in the gastrointestinal environment) and / or the condition of the subject (e.g., whether the subject can tolerate oral administration).

[0278] The exact amount of compound required to achieve an effective amount will depend, for example, on the species, age, and general condition of the subject; the severity of the side effects or symptoms; the nature of the specific compound; the mode of administration, etc., which varies from subject to subject. The desired dose may be delivered three times a day, twice a day, once a day, every other day, every three days, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dose may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations).

[0279] In certain embodiments, an effective amount of the compound for administration once or more per day to a 70 kg adult may comprise from about 0.0001 mg to about 3000 mg, from about 0.0001 mg to about 2000 mg, from about 0.0001 mg to about 1000 mg, from about 0.001 mg to about 1000 mg, from about 0.01 mg to about 1000 mg, from about 0.1 mg to about 1000 mg, from about 1 mg to about 1000 mg, from about 10 mg to about 1000 mg, or from about 100 mg to about 1000 mg of the compound per unit dosage form.

[0280] In certain embodiments, the compounds of the invention may be administered orally or parenterally at a dosage level sufficient to deliver from about 0.001 mg / kg to about 100 mg / kg, from about 0.01 mg / kg to about 50 mg / kg, preferably from about 0.1 mg / kg to about 40 mg / kg, preferably from about 0.5 mg / kg to about 30 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, and more preferably from about 1 mg / kg to about 25 mg / kg of the subject's body weight per day, once or multiple times a day to achieve a sustained therapeutic effect.

[0281] It should be understood that the dosage ranges described herein provide administration guidance for pharmaceutical compositions provided for adults. The amount to be administered to, for example, children or adolescents can be determined by a practitioner or one of ordinary skill in the art and may be lower or the same as the amount administered to adults.

[0282] It will also be appreciated that the compounds or compositions as described herein can be administered in combination with one or more additional therapeutic agents. The compounds or compositions can be administered in combination with additional therapeutic agents that increase their bioavailability, reduce and / or alter their metabolism, inhibit their excretion, and / or alter their distribution in the body. It will also be appreciated that the therapy employed can achieve the desired effect on the same condition and / or it can achieve different effects.

[0283] The compound or composition can be used simultaneously with one or more other therapeutically active agents, before or after them. In general, each agent will be used with a dosage and / or schedule determined for the agent. It will be further understood that the other therapeutically active agents used in this combination can be co-administered in a single composition or separately administered in different compositions. The specific combination used in the scheme will take into account the compatibility of the compounds of the invention with other therapeutically active agents and / or the desired therapeutic effect to be achieved. In general, it is expected that the use level of the additional therapeutically active agents used in combination will not exceed the level at which they are used alone. In some embodiments, the level used in combination will be lower than those levels used alone. Other therapeutically active agents include antibiotic agents, such as antibiotics that can be used to treat tuberculosis. Exemplary antibiotics include, but are not limited to, isoniazid, rifampicin, pyrazinamide, ethambutol and streptomycin.

[0284] The present invention also encompasses medicine kits (e.g., pharmaceutical packs). The medicine kits provided may include a pharmaceutical composition or compound of the present invention and a container (e.g., a vial, an ampoule, a bottle, a syringe and / or a dispenser package or other suitable container). In some embodiments, the medicine kit provided may optionally further include a second container, the second container comprising a pharmaceutical excipient for diluting or suspending the pharmaceutical composition or compound of the present invention. In some embodiments, the pharmaceutical composition or compound of the present invention provided in the container and the second container is combined to form a unit dosage form.

[0285] Uses and treatments

[0286] In another aspect, the present invention provides a method for treating a bacterial infection in a patient in need of such treatment, the method comprising administering an effective amount of a compound of formula I or a single stereoisomer or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound of formula I or a single stereoisomer or a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof. In certain embodiments, the effective amount is a therapeutically effective amount. In certain other embodiments, the effective amount is a prophylactic effective amount.

[0287] In some embodiments, the compounds of the present invention may be active against a wide range of both Gram-positive and Gram-negative organisms. In these and other embodiments, the compounds of the present invention may be used to treat infections and inhibit microbial growth. Thus, the compounds of the present invention may be used to treat humans and animals with a broad spectrum of bacterial infections, such as impetigo, pneumonia, bronchitis, pharyngitis, endocarditis, urinary tract infections, diabetic foot ulcers, gastrointestinal infections, and bacteremia. These bacterial infections can be caused by any of the following bacteria - Staphylococcus aureus, coagulase-negative staphylococci, methicillin-resistant Staphylococcus aureus, methicillin-resistant coagulase-negative staphylococci, enterococci, beta-hemolytic streptococci, viridans group streptococci, mycobacterium infections due to multidrug-resistant Mycobacterium tuberculosis and other atypical mycobacteria such as Mycobacterium intracellulare and Mycobacterium avium, and emerging Gram-negative pathogens such as Chryseobacterium meningosepticum, Chryseobacterium indologense and other Gram-negative pathogens such as Escherichia coli, Klebsiella, Proteus, Serratia, Citrobacter, Pseudomonas, Burkholderia, Brucella, Yersinia, Francisella, Coxiella, Chlamydia, Salmonella, Rickettsia, Shigella and Campylobacter.

[0288] In one embodiment, bacterial infection is tuberculosis. In certain embodiments, tuberculosis infection is mycobacterium tuberculosis infection. In certain embodiments, tuberculosis infection is multidrug resistant tuberculosis (MDR-TB) infection, for example, first-line TB medicine rifampicin and / or isoniazid are resistant. In certain embodiments, tuberculosis infection is extensively drug-resistant tuberculosis (XDR-TB) infection, for example, also three or more classes in six classes of second-line drugs are resistant (see, for example, Centers for Disease Control and Prevention (CDC) (2006). "Emergence of Mycobacterium tuberculosis with extensive resistance to second-line drugs worldwide, 2000-2004". MMWR MorbMortal Wkly Rep 55 (11): 301-5).

[0289] Technology

[0290] In some aspects, the compounds and intermediates of the present disclosure can be prepared according to the following general synthetic schemes I and II. In the general schemes, such as R 1 , R 2 , R 3 , R x , R y , Rx’ , R y’ The variables of , ring A, ring B, ring C, J, L, Y, m, n and p have the same definitions as in the preceding paragraph; X is a leaving group such as halide, mesylate, tosylate, triflate; and P is a hydroxy protecting group. In some embodiments, X is halide; and P is TBS.

[0291] General Synthesis Scheme I

[0292]

[0293] In general synthetic scheme I, ketone (a) undergoes reduction to produce alcohol (b), and protection of the hydroxyl group affords intermediate (c).

[0294] In step 3 of general synthetic scheme I, intermediate (c) is reacted with triisopropyl borate in the presence of a base such as butyl lithium to give boronate (d). In step 4, boronate (d) is cross-coupled with cytosine in the presence of a base such as a tertiary amine and a copper reagent such as a copper (II) reagent to give a compound of formula F.

[0295] In step 5 of general synthetic scheme I, the compound of formula F and the compound of formula (e) undergo amide coupling to produce intermediate (f). In some processes, about 1.1 to 2.0 molar equivalents of the compound of formula (e) are combined with 1 molar equivalent of the compound of formula F in a suitable solvent such as a polar aprotic solvent. Polar aprotic solvents include solvents such as dichloromethane, dimethylformamide, acetonitrile, and the like. The mixture in the polar aprotic solvent is then reacted at a temperature of about 0°C to 100°C for a sufficient time. In some embodiments, the temperature is about 25°C to 95°C or about 50°C to 95°C, and the reaction time is about 1 to 24 hours or 2 to 20 hours or about 5 to 18 hours.

[0296] In steps 6 and 7, the compound of formula (f) is deprotected to generate the free alcohol, which is then oxidized to the ketone, ie, the compound of formula A.

[0297] In step 8 of general synthetic scheme I, a compound of formula A is reacted with an amine under reductive amination conditions to obtain a compound of formula I. Reductive amination can be carried out in the presence of a reducing agent and a suitable solvent. Suitable solvents include protic solvents or aprotic solvents. Protic solvents include, but are not limited to, water and alcohols such as methanol, ethanol, propanol, and the like. Aprotic solvents include, but are not limited to, solvents such as dichloromethane, dimethylformamide, acetonitrile, and the like. Suitable solvents can also be a combination of two or three solvents. Reducing agents include, but are not limited to, borohydride reagents or metal hydride reagents. Non-limiting examples are lithium borohydride, sodium borohydride, and sodium cyanoborohydride.

[0298] General Synthesis Scheme II

[0299]

[0300] In step 1 of general synthetic scheme II, ketone (a) undergoes reductive amination with an amine to produce intermediate (g). The conditions for performing the reductive amination are provided in the previous paragraph.

[0301] In step 2 of general synthetic scheme II, intermediate (g) is reacted with triisopropyl borate in the presence of a base such as butyl lithium to give boronate (h). In step 3, boronate (d) is cross-coupled with cytosine in the presence of a base such as a tertiary amine and a copper reagent such as a copper (II) reagent to give a compound of formula III.

[0302] In step 4 of general synthetic scheme II, a compound of formula III and a compound of formula (e) undergo amide coupling to produce a compound of formula (I). The conditions for carrying out the amide coupling are similar to those in step 5 of general synthetic scheme I.

[0303] In one aspect, the present disclosure provides methods for preparing compounds of formula I:

[0304]

[0305] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, the process comprising:

[0306] The compound of formula A:

[0307]

[0308] and:

[0309] Formula B' (B') or formula C (C) Compound

[0310] Combined under reductive amination conditions to provide the compound of formula I,

[0311] Among them, ring A, ring B, ring C, J, L, R 1 , R 2 , R 3 , R x , R y , R x’ , R y’ , m, n and p are as defined in the preceding paragraph;

[0312] Ring B1 is a nitrogen-containing 3-8 membered monocyclic heterocycloalkylene or a nitrogen-containing 6-12 membered bicyclic heterocycloalkylene, each of which is optionally and independently substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2 and C1-C6 hydroxyalkyl;

[0313] Y is a bond or a C1-C6 alkylene optionally substituted by OH, NH2, CN, halo or C1-C6 alkoxy, wherein one methylene unit of the C1-C6 alkylene is optionally replaced by NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl) or N-(C1-C6 alkylene-C3-C8 cycloalkyl);

[0314] Y1 is a bond or C1-C5 alkylene optionally substituted with OH, NH2, CN, halo or C1-C6 alkoxy; and

[0315] R z It is H, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 haloalkyl or C1-C6 alkylene-C3-C8 cycloalkyl.

[0316] In some embodiments, Ring B1 is a nitrogen-containing 4-7 membered monocyclic heterocycloalkylene or a nitrogen-containing 6-9 membered bicyclic heterocycloalkylene, each of which is optionally substituted with up to three substituents independently selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

[0317] In some embodiments, Ring B1 is a nitrogen-containing 4-7 membered monocyclic heterocycloalkylene or a nitrogen-containing 6-9 membered bicyclic heterocycloalkylene. In some embodiments, Ring B1 is a nitrogen-containing 4-7 membered monocyclic heterocycloalkylene or a nitrogen-containing 6 membered bicyclic heterocycloalkylene.

[0318] In some embodiments, Ring B in Formula B' is selected from the group consisting of:

[0319] In some embodiments, Ring B1 is selected from the group consisting of:

[0320] In some embodiments, L is a bond, -CH2-, or -CH2-CH2-; and R x’ and R y’ Each is independently H, Boc or methoxycarbonyl.

[0321] In some embodiments, Y1 is a bond or a C1-C3 alkylene optionally substituted with OH, NH2, CN, halo, or C1-C6 alkoxy. In some embodiments, Y1 is a bond or a C1-C3 alkylene. In some embodiments, Y1 is a bond or -CH2-.

[0322] In some embodiments, R z is H, C1-C3 alkyl, C1-C3 hydroxyalkyl, C1-C3 haloalkyl or C1-C3 alkylene-C3-C6 cycloalkyl. z is H, Me, Et or CH2-cyclopropyl.

[0323] In some embodiments, the compound of Formula I is a compound of Formula IB, Formula IB′, or Formula IC:

[0324]

[0325] wherein variables are as defined herein.

[0326] In some embodiments, the compound of Formula I is a compound of Formula IB or Formula IC.

[0327] In some embodiments, the compound of Formula IB, Formula IB′, or Formula IC is selected from the compounds listed in Table 1 and Table 2.

[0328] In one aspect, the present disclosure provides methods for preparing compounds of formula IB:

[0329]

[0330] or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, the process comprising:

[0331] The compound of formula A is combined with the compound of formula B under reductive amination conditions to provide a compound of formula IB

[0332]

[0333] Among them, ring A, ring B, ring C, J, L, R 1 , R 2 , R 3 , R x , R y , R z , R x’ , R y’, m, n and p have the definitions provided in the preceding paragraph.

[0334] In one embodiment, the process further comprises when R x , R y , R x’ and R y’ When any one of the above is an amino protecting group, a step of removing the amino protecting group.

[0335] In another aspect, the present disclosure provides methods for preparing compounds of formula IC:

[0336]

[0337]

[0338] or a pharmaceutically acceptable salt thereof, the process comprising:

[0339] The compound of formula A is combined with the compound of formula C under reductive amination conditions to provide a compound of formula IC

[0340]

[0341] Among them, ring A, ring B1, ring C, J, L, R 1 , R 2 , R 3 , R x , R y , R x’ , R y’ , m, n and p have the definitions provided in the preceding paragraph.

[0342] In one embodiment, the process further comprises when R x , R y , R x’ and R y’ when at least one of the amino protecting groups is an amino protecting group, a step of removing the amino protecting group.

[0343] The procedures and conditions used to carry out the reductive amination of compounds of formula E are the same as in the general synthetic scheme.

[0344] In some embodiments, the reductive amination between the compound of formula A and the compound of formula B (or formula B') or between the compound of formula A and the compound of formula C can be carried out in the presence of a reducing agent and a suitable solvent. Suitable solvents include protic solvents or aprotic solvents. Protic solvents include, but are not limited to, water and alcohols, such as methanol, ethanol, propanol, etc. Aprotic solvents include, but are not limited to, solvents such as dichloromethane, dimethylformamide, acetonitrile, etc. A suitable solvent can also be a combination of two or three solvents. Reducing agents include, but are not limited to, borohydride reagents or metal hydride reagents. Non-limiting examples are lithium borohydride, sodium borohydride, sodium cyanoborohydride. In some processes, about 1.1 to 2.0 molar equivalents of the compound of formula B (or formula B') or C are combined with 1 molar equivalent of the compound of formula A and 1.0 to 2.0 molar equivalents of the reducing agent in a suitable solvent. The mixture is then reacted at a temperature of about 0°C to 100°C for a sufficient time. In some embodiments, the temperature is about 10°C to 95°C, or about 10°C to 50°C, or at room temperature, and the reaction time is about 1 to 24 hours, or 2 to 20 hours, or about 5 to 18 hours.

[0345] In some embodiments, the compound of formula A is a compound of formula ID

[0346]

[0347] In another aspect, the present disclosure provides a process for preparing a compound of formula ID or a pharmaceutically acceptable salt thereof, the process comprising:

[0348] The compound of formula E

[0349]

[0350] Coupling with a compound of formula F

[0351]

[0352] Among them, ring C, K, R 1 , R 2 , R 3 , R x , R y , m, n and p are as defined herein; and

[0353] P is a hydroxy protecting group.

[0354] The process and conditions used to carry out the amide coupling of the compound of formula E with the compound of formula F are as described in step 5 of general synthetic scheme I.

[0355] In one embodiment, the process further comprises the step of removing the hydroxy protecting group to provide a compound of formula IE

[0356]

[0357] In some embodiments, the compound of Formula IE is selected from the group consisting of:

[0358]

[0359] In another embodiment, the process further comprises the step of oxidizing the hydroxyl group in Formula IE to provide a compound of Formula ID. Exemplary oxidizing agents include Dess-Martin periodinane, chromium trioxide, pyridinium chlorochromate, dimethyl sulfoxide, oxalyl chloride, and the like.

[0360] In one embodiment, the compound of formula ID is a compound of formula ID-1a or ID-1b:

[0361]

[0362] or a pharmaceutically acceptable salt thereof, wherein X1, K, R 1 , R 2 , R 3 , R x , R y , m, n and p have the definitions provided in any of the preceding paragraphs.

[0363] In another embodiment, the compound of formula ID is a compound of formula ID-2a or ID-2b:

[0364]

[0365] or a pharmaceutically acceptable salt thereof, wherein R x , R y , X1 and K have the definitions provided in the preceding paragraph.

[0366] In another embodiment, the compound of formula ID is selected from the group consisting of:

[0367]

[0368]

[0369] Compound preparation

[0370] The preparation of starting materials which are commercially available, described in the literature or readily available to one skilled in the art is not described. One skilled in the art will appreciate that where compounds are stated to be prepared in a manner similar to the earlier examples or intermediates, reaction times, equivalents of reagents and temperatures may be modified for each specific reaction and it may be necessary or desirable to employ different post-processing or purification techniques. Where microwave irradiation was used to carry out the reaction, the microwave oven used was either a Biotage Initiator or a CEM Discover System Model 908005. The actual power supplied was varied during the course of the reaction to maintain a constant temperature.

[0371] General Methods

[0372] Reactions requiring anhydrous conditions were all performed in oven-dried glassware under a positive pressure of nitrogen or argon. Commercially available reagents were used as received; otherwise, materials were purified according to the purification of laboratory chemicals. Dichloromethane (CH2Cl2), N,N'-dimethylformamide (DMF), toluene, and tetrahydrofuran (THF) were degassed with nitrogen and passed through a solvent purification system (Innovative Technologies Pure Solv). Anhydrous 1,4-dioxane was purchased from Acros Organics and purified at Acros Seal TM Triethylamine (Et3N) and N,N-diisopropylethylamine (DIPEA) were stored in On molecular sieve or The HPLC-MS / MS was performed on a 100 nm column (100 nm, 100 nm, 0.1 nm, 0.2 nm, 1.3 nm, 0.5 nm, 1.4 nm, 0.6 nm, 1.8 nm, 2.5 nm, 1.9 nm, 2.3 nm, 1.6 nm, 2.7 nm, 1.8 nm, 2.9 nm, 1.9 nm, 2.5 nm, 1.8 nm, 1.9 nm, 2.5 nm, 1.8 nm, 1.9 nm, 2.5 nm, 1.8 nm, 2.5 ...

[0373] use Melting points were determined using a capillary melting point apparatus. Infrared spectra were obtained using a Nicolet 380-FT IR spectrometer equipped with a Smart Orbit sample system. Optical rotations were obtained at ambient temperature on a Perkin Elmer 343 polarimeter (Na D line) using a microcell with an optical path length of 1 decimeter. Mass spectra determined by LCMS were obtained on a Thermo Scientific TM UltiMate TM The data were acquired on a Waters Aquity UPLC H-Class series with a photodiode array detector and a QDa mass detector. 1 H NMR spectra were recorded at 500 MHz, 400 MHz and 300 MHz, and 13 C was recorded at 125 MHz. Proton resonances are reported relative to the deuterated solvent peaks: 7.27 ppm for CDCl3, 3.31 ppm for CD3OD (centerline signal), 2.50 for D6-DMSO and 4.79 for D2O using the following format: Chemical shift (δ) [multiplicity (s=singlet, br s=broad singlet, d=doublet, t=triplet, q=quartet, m=multiplet)]. Carbon resonances are reported as chemical shifts (δ) in parts per million relative to the centerline signal of the corresponding solvent peaks: 77.23 ppm for CDCl3 and 49.15 ppm for CD3OD. Commercially available chemicals were purchased from multiple suppliers, including Sigma-Aldrich, Acros, Enamine, TCI America, Combi-Blocks, Alfa-Aesar, Angene, Ark Pharma, PharmaBlock, Strem Chemicals, Frontier Scientific, and AstaTech, Inc.

[0374] Liquid chromatography-mass spectrometry

[0375] Liquid chromatography-mass spectrometry A

[0376] Total ion current (TIC) and DAD UV chromatographic traces as well as peak-related MS and UV spectra were analyzed on a UPLC / MS Acquity equipped with a PDA detector and coupled to a Waters single quadrupole mass spectrometer operating in alternating positive and negative electrospray ionization mode. TM [LC / MS-ES(+ / -): using Acquity UPLC TMAnalysis by CSH, C18 column (50 x 2.1 mm, 1.7 μm particle size), column temperature 40°C, mobile phase: A-water + 0.1% HCOOH / B-CH3CN + 0.1% HCOOH, flow rate: 1.0 mL / min, run time = 2.0 min, gradient: t = 0 min 3% B, t = 1.5 min 99.9% B, t = 1.9 min 99.9% B, t = 2.0 min 3% B, stop time 2.0 min. Positive ES 100-1000, negative ES 100-1000, UV detection DAD 210-350 nm.

[0377] Liquid chromatography-mass spectrometry B

[0378] Total ion current (TIC) and DAD UV chromatographic traces as well as peak-related MS and UV spectra were analyzed on a UPLC / MS Acquity equipped with a PDA detector and coupled to a Waters single quadrupole mass spectrometer operating in alternating positive and negative electrospray ionization mode. TM [LC / MS-ES(+ / -): using Acquity UPLC TM Analysis by BEH, C18 column (50×2.1 mm, 1.7 μm particle size), column temperature 40° C., mobile phase: A-0.1% v / v aqueous ammonia solution (aq) pH 10 / B-CH3CN, flow rate: 1.0 mL / min, run time = 2.0 minutes, gradient: t=0 minutes 3% B, t=1.5 minutes 99.9% B, t=1.9 minutes 99.9% B, t=2.0 minutes 3% B, stop time 2.0 minutes. Positive ES 100-1000, negative ES 100-1000, UV detection DAD 210-350 nm.

[0379] Liquid chromatography-mass spectrometry C

[0380] LC / MS-ES (+ / -): Analysis performed using AQUITY with PDA detector and QDA Performance, C18 column (50×2.1 mm, 1.6 μm particle size), column temperature 35°C, mobile phase: A-0.1% formic acid in Milli Q water (pH=2.70) / B-0.1% formic acid in water:acetonitrile (10:90), flow rate: 0.8-1.0 mL / min, run time = 4.0 minutes, gradient: t=0 minutes 3% B, t=2.7 minutes 98% B, t=3.0 minutes 100% B, t=3.51 minutes 3% B, stop time 4.0 minutes.

[0381] Liquid chromatography-mass spectrometry D

[0382] LC / MS-ES (+ / -): Analysis performed using AQUITY H-Class with PDA detector and QDA, C18 column (50×2.1 mm, 1.6 μm particle size), column temperature 35°C, mobile phase: A-0.1% formic acid in Milli Q water (pH=2.70) / B-0.1% formic acid in water:acetonitrile (10:90), flow rate: 0.8-1.0 mL / min, run time = 4.0 minutes, gradient: t=0 minutes 3% B, t=2.7 minutes 98% B, t=3.0 minutes 100% B, t=3.51 minutes 3% B, stop time 4.0 minutes.

[0383] Liquid chromatography-mass spectrometry E

[0384] LC / MS-ES (+ / -): Analysis performed using AQUITY H-Class with PDA detector and QDA, C18 column (50×2.1 mm, 1.6 μm particle size), column temperature 35°C, mobile phase: A-0.1% formic acid in water (pH=2.70) / B-0.1% formic acid in water:acetonitrile (10:90), run time = 9.0 minutes, gradient: t=0 minutes 1% B, t=2.5 minutes 50% B, t=4.5 minutes 97.5% B, t=6.5 minutes 1% B, stop time 9.0 minutes.

[0385] Liquid chromatography-mass spectrometry F

[0386] LC / MS-ES (+ / -): Analysis was performed using an Agilent Infinity II G6125C LCMS, C18 column (50×4.6 mm, 3.5 μm particle size), column temperature 35°C, mobile phase: A-5 mM ammonium bicarbonate (pH=7.35) in Milli-Q water / B-methanol, run time = 7.0 min, gradient: t=0 min 8% B, t=3.0 min 70% B, t=3.7 min 95% B, t=4.2 min 100% B, t=5.21 min 8% B, stop time 7.0 min.

[0387] Liquid chromatography-mass spectrometry G

[0388] LC / MS-ES (+ / -): Analysis was performed using a Waters Alliance 2690 with Micromass ZQ and a 996PDA detector, a C18 column (150×4.6 mm, 3.5 μm particle size), a column temperature of 35°C, a mobile phase: A-5 mM ammonium acetate in water + 0.1% FA / B-methanol, run time = 17.0 minutes, a gradient: t=0 minutes 10% B, t=7.0 minutes 90% B, t=9.0 minutes 100% B, t=14.01 minutes 10% B, a stop time of 17.0 minutes.

[0389] Liquid chromatography-mass spectrometry H

[0390] LC / MS-ES (+ / -): Analysis performed using AQUITY with PDA detector and QDA Performance, C18 column (50×2.1 mm, 1.6 μm particle size), column temperature 35°C, mobile phase: A-0.1% formic acid in Milli Q water (pH=2.70) / B-0.1% formic acid in water:acetonitrile (10:90), flow rate: 0.9 mL / min, run time = 3.0 minutes, gradient: t=0 minutes 5% B, t=1.8 minutes 98% B, t=2.0 minutes 100% B, t=2.51 minutes 5% B, stop time 17.0 minutes.

[0391] Analytical methods

[0392] 1 H nuclear magnetic resonance (NMR) spectroscopy was performed using one of the following instruments: a Bruker Avance 400 instrument equipped with a probe DUAL 400 MHz S1, a Bruker Avance 400 instrument equipped with a probe 6 S1 400 MHz 5 mm 1 H- 13 A Bruker Avance 400 instrument with C ID, a Bruker Avance III 400 instrument with nanobay equipped with a broadband BBFO 5 mm direct probe, a 400 MHz Agilent direct drive instrument equipped with an ID AUTO-X PFG probe (all operated at 400 MHz), or a Bruker Avance III 400 instrument with a 5 mm triplet resonant probe. 1 H{ 13 C / 15 N} Agilent VNMRS500 direct drive instrument (operating at 500 MHz) with cryoprobe. Unless otherwise stated, spectra were obtained at about room temperature in the stated solvents. In all cases, the NMR data are consistent with the proposed structure. Characteristic chemical shifts (δ) are given in parts per million using conventional abbreviations for designating major peaks: for example, s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; dt, doublet of triplets; br, broad.

[0393] Thin layer chromatography (TLC) refers to silica gel TLC using silica gel F254 (Merck) plates. fIt is the distance travelled by the compound divided by the distance travelled by the solvent on the TLC plate. Column chromatography was performed on Biotage silica gel columns (KP-Sil or KP-NH) using an automated flash chromatography (Biotage SP1 or Isolera) system or on a Biotage C18 column (KP-C18) in the case of reverse phase chromatography.

[0394] Preparative HPLC was performed on Shimadzu LC-20AP, Waters 2545 and Agilent 1260 infinity. Purity was determined on Waters Alliance e2695-PDA detector 2998 and Agilent 1260 Infinity-II. (Mobile phase: 0.05% HCl / methanol in water in a gradient elution method).

[0395] Table 5. Abbreviations and names of reagents

[0396]

[0397]

[0398] Intermediate synthesis

[0399]

[0400] 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-

[0401] Imidazol-3-ium iodide (Intermediate 1)

[0402] Solution 1

[0403]

[0404] Reagents: 1) 2-((tert-Butyloxycarbonyl)amino)-2-methylpropanoic acid, HATU, DIPEA, DMF, room temperature, 16 hours 2) 10% Pd / C, CH3OH, room temperature, 16 hours 3) CDI, CH2Cl2, room temperature, 16 hours 4) MeI, CH3CN, room temperature, 16 hours.

[0405] Step 1: Benzyl 4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxylate. To a stirred solution of 2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (35.5 g, 175 mmol) in DMF (350 mL) at 0°C was added DIPEA (51.2 g, 397 mmol) and HATU (90.6 g, 238 mmol). The reaction mixture was stirred at 0°C for 45 minutes. Benzyl piperazine-1-carboxylate (35 g, 158.9 mmol) was added to the reaction mixture at 0°C and stirred at room temperature for 16 hours. The resulting reaction mixture was poured into H2O (1500 mL) and extracted with EtOAc (3 x 700 mL), and the combined organics were dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude material was purified by flash chromatography (EtOAc:Hex) to give the desired product as an off-white solid (36 g, 55%) LCMS [M+H] 406.

[0406] Step 2: tert-Butyl (2-methyl-1-oxo-1 (piperazin-1-yl) propan-2-yl) carbamate. To a stirred solution of benzyl 4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxylate (35 g, 86.4 mmol) in CH3OH (500 mL) was added 10% Pd / C (3.5 g). The reaction mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. The resulting reaction mixture was purified by Filter and wash with CH3OH (1500 mL). The filtrate was concentrated under reduced pressure and dried to give the desired product (25.0 g, 100%) as a viscous oil. LCMS [M+H] 272.

[0407] Step 3: tert-butyl (1-(4-1H-imidazole-1-carbonyl)piperazine-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. To a stirred solution of tert-butyl (2-methyl-1-oxo-1 (piperazine-1-yl)propan-2-yl)carbamate (25 g, 92.2 mmol) in CH2Cl2 (300 mL) was added CDI (17.78 g, 110 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. The resulting crude material was purified by flash chromatography (CH3OH:CH2Cl2) to give the desired product (30.0 g, 89%) as an off-white solid. 1H NMR (DMSO-d6, 400MHz): δ8.04(s,1H),7.48(s,1H),7.36(s,1H),7.03(s,1H),3.65-3.52(m,4H),3.51-3.40(m,4H),1.38(s,6H),1.30(s,9H). LCMS[M+H]366.3.

[0408] Step 4: 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide. To a stirred solution of tert-butyl (1-(4-1H-imidazole-1-carbonyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate (20 g, 55 mmol) in CH3CN (250 mL) was added MeI (20.8 ml, 329 mmol) at 0°C. The reaction mixture was stirred at room temperature for 16 hours and then concentrated under reduced pressure to give the desired product (30 g, quantitative) as a pale yellow solid. 1 H NMR(DMSO-d6,400MHz): δ9.57 9s,1H),8.05(s,1H),7.87(t,1H),7.40(s,1H),3.93(s,3H),3.78-3.65(m,4H),3.59-3.45(m,4H),1.40(s,6H),1.32(S,9H). LCMS[M+H]380.2.

[0409]

[0410] (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalene-2-

[0411] tert-Butyl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate (Intermediate 2)

[0412] Solution 2

[0413]

[0414] Reagents: 1) NaBH4, CH3OH, room temperature, 3 hours 2) TBDMSCl, imidazole, CH2Cl2, room temperature, 16 hours 3) n-BuLi, B(OiPr)3, THF, -78°C to room temperature, 3 hours 4) Cytosine, TMEDA, Cu(OAc)2·H2O, CH3OH:H2O (4:1), O2, room temperature, 16 hours 5) 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide, CH3CN, 90°C, 16 hours 6) MeOH, TsOH, room temperature, 16 hours 7) DMP, CH2Cl2, room temperature, 3 hours.

[0415] Step 1: 6-bromo-1,2,3,4-tetrahydronaphthalene-2-ol. To a stirred solution of 6-bromo-3,4-dihydronaphthalene-2(1H)-one (10 g, 44 mmol) in MeOH (250 mL) was added NaBH4 (1.7 g, 44 mmol) at 0 ° C. The reaction mixture was stirred at room temperature for 3 hours, and the volatiles were removed under reduced pressure. The crude solid was dissolved in EtOAc (500 mL), and the reaction mixture was washed with H2O (1x500 mL). The organic matter was dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography to obtain the desired product (9.0 g, 90%) as a brown oil.

[0416] Step 2: ((6-bromo-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)(tert-butyl)dimethylsilane. To a stirred solution of 6-bromo-1,2,3,4-tetrahydronaphthalen-2-ol (49.0 g, 44 mmol) in CH2Cl2 (500 mL) was added imidazole (3.2 g, 48 mmol) and tert-butyldimethylsilyl chloride (7.2 g, 48 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 16 hours. The resulting reaction mixture was poured into H2O (500 mL) and extracted with CH2Cl2 (3×700 mL). The combined organics were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (EtOAc:Hex) to give the desired product (85%) as a yellow oil. 1 H NMR(500MHz, CDCl3)δ7.24(s,1H),7.22(d,1H),6.93(d,1H),4.15-4.06(m,1H),2.99-2.88(m,2H) ,2.80-2.73(m,1H),2.69(dd,1H),1.98-1.91(m,1H),1.84-1.73(m,1H),0.92(s,9H),0.12(d,6H).

[0417] Step 3: Diisopropyl (6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)borate. To a stirred solution of ((6-bromo-1,2,3,4-tetrahydronaphthalen-2-yl)oxy)(tert-butyl)dimethylsilane (3.7 g, 9.5 mmol) in THF (300 mL) was added n-BuLi (2.5 M in hexanes, 4.1 mL, 10.4 mmol) at -78 °C under N2 atmosphere. The reaction mixture was stirred at -78 °C for 30 minutes. Triisopropyl borate (2.4 mL, 10.4 mmol) was added at -78 °C. The reaction mixture was warmed to room temperature and stirred for 3 hours. The resulting reaction mixture was poured into NH4Cl solution (100 mL) and extracted with EtOAc (3×300 ml). The combined organics were dried over Na2SO4, filtered and concentrated under reduced pressure to give the desired product (90%) as a clear oil.

[0418] Step 4: 4-amino-1-(6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)pyrimidin-2(1H)-one. A solution of diisopropyl (6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)borate (9.9 g, 25 mmol) and cytosine (3.1 g, 28 mmol) in MeOH:H2O (500 mL, 4:1) was stirred at room temperature for 30 minutes in an air-exposure state. TMEDA (5.1 mL, 34 mmol) and Cu(OAc)2.H2O (5.6 g, 28 mmol) were added, and the reaction mixture was stirred at room temperature in an air-exposure state for 48 hours. The reaction mixture was concentrated under reduced pressure and cold H2O (100 mL) was added to the mixture. The solid was filtered and washed with H2O (3 x 100 mL) and Et2O (2 x 60 mL) under reduced pressure. The resulting solid was dried to give the desired product (5.5 g, 60%) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ7.56(d,1H),7.29-7.08(m,3H),7.03(d,2H),5.75(d,1H),4.15(s,1H),2.97(dd,1H),2.9 2-2.82(m,1H),2.81-2.71(m,1H),2.66(dd,1H),1.98-1.85(m,1H),1.78-1.62(m,1H),0.87(s,9H),0.09(d,6H). LCMS[M+H]372.2.

[0419] Step 5: tert-Butyl (1-(4-((1-(6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. A mixture of 4-amino-1-(6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)pyrimidin-2(1H)-one (1.5 g, 3.5 mmol) and 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide (Intermediate 2, 3.1 g, 5.3 mmol) in CH3CN (45 mL) was heated to 90°C for 16 hours. The reaction mixture was concentrated under reduced pressure and the crude material was purified by column chromatography (MeOH:CH2Cl2) to give the title compound (1.67 g, 80%) as a white solid. 1 H NMR (500MHz, CDCl3) δ12.89(s,1H),7.22(d,1H),7.12(d,1H),7.03(d,1H),7.02(s,1H),5. 77(d,1H),5.02(br.s,1H),4.17-4.02(m,1H),3.83(br.s,2H),3.74(br.,2H),3.69(br.s, 2H),3.58(br.s,2H),3.44(s,1H),2.99-2.86(m,2H),2.81-2.66(m,2H),1.96-1.86(m,1H) ,1.84-1.71(m,1H),1.47(s,6H),1.40(s,9H),1.27-1.15(m,1H),0.87(s,9H),0.07(s,6H). LCMS[M+H]669.4.

[0420] Step 6: tert-Butyl (l-(4-((l-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-l-yl)-2-methyl-l-oxopropan-2-yl)carbamate. To a stirred solution of tert-butyl (l-(4-((l-(6-((tert-butyldimethylsilyl)oxy)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-l-yl)-2-methyl-l-oxopropan-2-yl)carbamate (300 mg, 0.45 mmol) in MeOH (10 mL) at 0 °C was added TsOH (172 mg, 0.90 mmol). The reaction mixture was stirred at room temperature for 16 hours, then poured into aqueous NaHCO3 (25 mL) and extracted with CH2Cl2 (3×50 mL). The combined organics were dried over Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (MeOH:CH2Cl2) to give the desired product (290 mg, 93%) as an off-white solid. LCMS [M+H] 555.3.

[0421] Step 7: tert-Butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate. To a stirred solution of tert-butyl (1-(4-((1-(6-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate (290 mg 0.42 mmol) in DCM (5 mL) at room temperature was added Dess-Martin periodinane (259 mg, 0.63 mmol). The reaction mixture was stirred at room temperature for 3 hours, poured into saturated NaHCO3 (20 mL) and extracted with CH2Cl2 (3×50 mL). The combined organics were dried over Na2SO4, filtered and concentrated under reduced pressure below 35°C to give the desired product (258 mg, quantitative) as an off-white solid. LCMS [M+H] 553.3.

[0422]

[0423] 4-((2R,4S)-2-(tert-butyl)-4-methyloxazolidine-4-carbonyl)-N-(2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide (Intermediate 3)

[0424] Prepared from 1-(4-((2R,4S)-2-(tert-butyl)-4-methyloxazolidin-4-carbonyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide and 4-amino-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)pyrimidin-2(1H)-one as in Scheme 2. LCMS [M+H] 537.3.

[0425]

[0426] Tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(2-oxo-2,3-dihydro-1H-inden-5-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate (Intermediate 4)

[0427] Prepared from benzyl 4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxylate and 4-amino-1-(2-((tert-butyldimethylsilyl)oxy)-2,3-dihydro-1H-inden-5-yl)pyrimidin-2(1H)-one as in Scheme 2. LCMS [M+H] 541.2.

[0428] Compound synthesis

[0429]

[0430] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 9)

[0431] Solution 3

[0432]

[0433] Reagents: 1) tert-butyl (1S, 3S)-3-aminocyclopentyl)carbamate, Na(OAc)3BH, DCE, room temperature, 16 hours 2) HCl / MeOH, room temperature, 4 hours.

[0434] Step 1: tert-Butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. To a solution of tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate (8.5 g, 15 mmol) in DCE (500 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (3.4 g, 17 mmol), Na(OAc)3BH (4.9 g, 23 mmol), and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was treated with aqueous NaHCO3 (500 mL), the phases were separated, and the aqueous layer was extracted with CH2Cl2 (3×500 mL). The combined organics were dried over Na2SO4, concentrated under reduced pressure and purified by column chromatography (CH2Cl2:MeOH:NH4OH) to give the desired compound (75%) as a grey solid. 1 H NMR(500MHz, CDCl3)δ7.22(d,1H),7.15(d,1H),7.05(d,1H),7.04(s,1H),5.79(d,1H),4.8 6(br.s,1H),4.49(br.s,1H),4.08(br.s,1H),3.86(br.s,2H),3.76(br.s,2H),3.71(br.s, 2H),3.61(s,2H),3.48(br.s,1H),3.13-2.98(m,2H),2.96-2.75(m,2H),2.71-2.56(m,1H), 2.24-1.98(m,3H),1.91-1.74(m,2H),1.64(s,1H),1.51(s,6H),1.43(d,20H),1.28(s,1H). LCMS[M+H]737.8.

[0435] Step 2: 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. Tert-butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate was dissolved in a solution of HCl / MeOH (5 mL) and stirred for 4 hours. The volatiles were removed under reduced pressure and the crude solid was purified by reverse phase HPLC (H2O:CH3CN:TFA) and concentrated under reduced pressure.Addition of HCl / MeOH (3x15 mL) and evaporation under reduced pressure gave the desired product. 1 H NMR(400MHz,D2O)δ7.76(d,1H),7.20(d,1H),7.09(d,2H),6.68(d,1H),4.03-3.93(m,1H),3.83-3.73(m,1H), 3.63(s,4H),3.58(s,6H),3.23(d,1H),2.88-2.77(m,3H),2.33-2.12(m,5H),1.83-1.62(m,2H),1.59(s,6H). LCMS[M+H]537.2.

[0436]

[0437] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((1R,5S,6s)-6-amino-3-azabicyclo[3.1.0]hex-3-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 1)

[0438] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1R,5S,6s)-3-azabicyclo[3.1.0]hexan-6-yl)carbamate. 1H NMR(400MHz,D2O)δ7.87(d,1H),7.30(d,1H),7.21(s,2H),6.80(d,1H),4.01(d,2H),3.71(d,10H) ,3.33(s,1H),3.09-2.87(m,4H),2.83(s,1H),2.42(s,2H),2.34(s,1H),1.89(m,1H),1.70(d,6H). LCMS[M+H]535.2.

[0439]

[0440] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(4-aminopiperidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 2)

[0441] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl piperidin-4-ylcarbamate. 1 H NMR(400MHz,D2O)δ8.02(d,1H),7.39(d,1H),7.28(s,1H),7.27(d,1H),6.82(d,1H),3.80(s,6H),3.77-3.71(m,5H),3.68 -3.57(m,1H),3.42-3.30(m,3H),3.25-3.15(m,1H),3.15-2.94(m,2H),2.51-2.33(m,3H),2.10-1.90(m,3H),1.75(s,6H). LCMS[M+H]537.3.

[0442]

[0443] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((1R,5S,6s)-6-(aminomethyl)-3-azabicyclo[3.1.0]hex-3-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 3)

[0444] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (((1R,5S,6r)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate. 1 H NMR(400MHz,D2O)δ7.95(d,1H),7.31(d,1H),7.22(s,2H),6.78(d,1H),3.91(d,2H),3.75(s,2H),3.71(s,6H),3.66-3. 56(m,2H),3.36(d,1H),3.08-2.86(m,6H),2.35(s,1H),2.04(d,2H),1.94-1.83(m,1H),1.70(d,6H),1.39-1.30(m,1H). LCMS[M+H]549.3.

[0445]

[0446] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((S)-3-(aminomethyl)pyrrolidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 4)

[0447] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (R)-(pyrrolidin-3-ylmethyl)carbamate. 1 HNMR(400MHz,D2O)δ7.94(d,1H),7.33(d,1H),7.22(d,2H),6.79(d,1H),4.06-3. 85(m,1H),3.87-3.27(m,12H),3.25-2.65(m,7H),2.56-2.29(m,2H),2.52-2.30(m 2H),1.71(s,6H). LCMS[M+H]537.3.

[0448]

[0449] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((R)-3-(aminomethyl)pyrrolidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 5)

[0450] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (S)-(pyrrolidin-3-ylmethyl)carbamate. 1 HNMR(400MHz,D2O)δ8.00(d,1H),7.34(d,1H),7.23(d,2H),6.78(d,1H),4.05-3.87(m,1H) ,3.86-3.27(m,12H),3.27-2.83(m,7H),2.81-2.24(m,2H),2.07-1.72(m,2H),1.70(s,6H). LCMS[M+H]537.4.

[0451]

[0452] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((cis)-4-aminocyclohexyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 6)

[0453] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((cis)-4-aminocyclohexyl)carbamate. 1 HNMR(400MHz,D2O)δ7.92-7.84(m,1H),7.34(d,1H),7.25(s,2H),6.85(d,1H),3.89-3.58(m,11H),3.43-3.33 (m,1H),3.07-2.94(m,3H),2.44-2.31(m,1H),2.17-2.07(m,2H),1.98(s,4H),1.85-1.78(m,1H),1.74(s,8H). LCMS[M+H]551.3.

[0454]

[0455] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 7)

[0456] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((trans)-4-aminocyclohexyl)carbamate. 1 HNMR(400MHz,D2O)1H NMR(400MHz,D2O)δ7.91(d,1H),7.35(d,1H),7.25(d,2H),6.84(d,1H),3.89-3.69(m,9H),3.50(s,1H),3.40- 3.23(m,2H),3.09-2.92(m,3H),2.32(s,3H),2.23(s,2H),1.98-1.85(m,1H),1.75(s,6H),1.68-1.53(m,4H). LCMS[M+H]551.3.

[0457]

[0458] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((6-aminospiro[3.3]hept-2-yl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 8)

[0459] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (6-aminospiro[3.3]hept-2-yl)carbamate. 1HNMR(400MHz,D2O)δ7.90(d,1H),7.34(d,1H),7.25(s,2H),6.84(d,1H),3.99(s,1H),3.86-3.68(m,8H),3.64(s,1H) ,3.33-3.23(m,1H),2.99(s,3H),2.71-2.56(m,2H),2.56-2.40(m,2H),2.39-2.21(m,6H),1.91(s,1H),1.75(s,6H). LCMS[M+H]563.3.

[0460]

[0461] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1R,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 10)

[0462] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1S,3R)-3-aminocyclopentyl)carbamate. 1 H NMR(400MHz,D2O)δ7.89(d,1H),7.36(d,1H),7.25(d,2H),6.85(d,1H),4.06-3.95(m,1H),3.85-3.68(m,10H),3.39 (d,1H),3.07-2.93(m,3H),2.82-2.71(m,1H),2.42-2.21(m,3H),2.05-1.89(m,3H),1.89-1.78(m,1H),1.75(s,6H). LCMS[M+H]537.3.

[0463]

[0464] 4-((S)-2-amino-3-hydroxy-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 24)

[0465] Prepared as in Scheme 3 from 4-((2R,4S)-2-(tert-butyl)-4-methyloxazolidine-4-carbonyl)-N-(2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate. 1 H NMR(400MHz,D2O)δ7.89-7.84(m,1H)7.36(d,1H),7.25(d,2H),6.86(d,1H),4.16(t,2H),3.92(d,2H),3.79(s,4H),3. 73(s,5H),3.37(s,1H),3.02(s,3H),2.41(d,2H),2.37-2.28(m,3H),1.99-1.86(m,1H)1.87-1.85(m,2H),1.70(s,3H). LCMS[M+H]553.5.

[0466]

[0467] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1R,3R)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 25)

[0468] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1R,3R)-3-aminocyclopentyl)carbamate. 1 H NMR(400MHz,D2O)δ7.97(d,1H),7.37(d,1H),7.26(s,1H),7.26(d,1H),6.83(d,1H),4.22-4.06(m,1H),4.01-3.86(m,1 H),3.79(s,4H),3.74(s,6H),3.44-3.33(m,1H),3.07-2.93(m,3H),2.53-2.27(m,5H),2.01-1.76(m,2H),1.75(s,6H). LCMS[M+H]537.4.

[0469]

[0470] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3R)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 26)

[0471] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1R,3S)-3-aminocyclopentyl)carbamate. 1 H NMR(400MHz,D2O)δ8.03(d,1H),7.37(d,1H),7.26(d,2H),6.82(d,1H),4.06-3.96(m,1H),3.80(s,5H),3.75(s,6H),3.44 -3.33(m,1H),3.08-2.95(m,3H),2.85-2.72(m,1H),2.42-2.19(m,3H),2.05-1.92(m,2H),1.89-1.79(m,1H),1.75(s,6H). LCMS[M+H]537.4.

[0472]

[0473] 4-(2-amino-2-methylpropanoyl)-N-(1-((S)-6-(((1S,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 27 and Compound 28)

[0474] Tert-butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate was separated by chiral HPLC to give optically pure stereoisomers.

[0475]

[0476] First eluting diastereomer 1: 1H NMR (400MHz, CDCl3) δ7.22(d,1H),7.15(d,1H),7.06(d,1H),7.05(s,1H),5.79(d,1H),4.85(br. s,1H),4.48(br.s,1H),4.07(br.s,1H),3.86(br.s,2H),3.75(br.s,2H),3.71(br.s,2H),3.61(s ,2H),3.48-3.38(m,1H),3.08-2.95(m,2H),2.95-2.77(m,2H),2.58(dd,1H),2.22-1.99(m,3H), 1.85-1.69(m,2H),1.63-1.54(m,1H),1.51(s,6H),1.44(d,18H),1.41-1.30(m,2H),1.25(s,1H). LCMS[M+H]737.8.

[0477] Compound 27 was prepared in an analogous manner to step 2 in Scheme 3 from optically pure first eluting diastereomer 1(1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamic acid tert-butyl ester. 1 H NMR(500MHz,D2O)δ7.97(d,1H),7.38(d,1H),7.28(s,1H),7.26(d,2H),6.85(d,1H),4.23-4.11(m,1H),4.01-3.90(m,1H),3.88- 3.68(m,9H),3.40(dd,1H),3.15-2.96(m,3H),2.54-2.37(m,3H),2.35(t,2H),2.00-1.91(m,1H),1.90-1.78(m,2H),1.76(s,6H). LCMS[M+H]537.4.

[0478] Second eluting enantiomer 2: 1H NMR(400MHz, CDCl3)δ7.22(d,1H),7.15(d,1H),7.05(dd,2H),5.78(d,1H),4.86(br.s,1H),4 .48(br.s,1H),4.07(br.s,1H),3.85(br.s,2H),3.75(br.s,2H),3.71(br.s,2H),3.61(br.s, 2H),3.47-3.39(m,1H),3.07-2.95(m,2H),2.95-2.77(m,2H),2.57(dd,1H),2.20-1.97(m,3H ),1.75(t,2H),1.64-1.54(m,1H),1.51(s,6H),1.43(d,18H),1.40-1.31(m,2H),1.26(s,1H). LCMS[M+H]737.8.

[0479] Compound 28 was prepared in an analogous manner to step 2 in Scheme 3 from the optically pure second eluting diastereomer 2(1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamic acid tert-butyl ester. 1 H NMR(500MHz,D2O)δ7.90(d,1H),7.33(d,1H),7.22(s,2H),6.80(d,1H),4.14-4.06(m,1H),3.93-3.86(m,1H),3.83-3.64(m ,10H),3.37(dd,1H),3.06-2.91(m,3H),2.45-2.31(m,4H),2.28(t,2H),1.96-1.85(m,1H),1.86-1.73(m,1H),1.71(s,6H). LCMS[M+H]537.4.

[0480]

[0481] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((trans-3-aminocyclobutyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 29)

[0482] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (trans-3-aminocyclobutyl)carbamate. 1 HNMR(400MHz,D2O)δ7.86(d,1H),7.35(d,1H),7.25(d,1H),7.24(d,1H),6.86(d,1H),4.41-4.29(m,1H),4.17-4.06(m,1H),3.79( br.s,3H),3.73(br.s,6H),3.31(dd,1H),3.05-2.93(m,3H),2.90-2.70(m,4H),2.36-2.26(m,1H),2.00-1.89(m,1H),1.75(s,6H). LCMS[M+H]523.4.

[0483]

[0484] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((cis-3-aminocyclobutyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 30)

[0485] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (cis-3-aminocyclobutyl)carbamate. 1 HNMR(400MHz,D2O)δ7.88(d,1H),7.35(d,1H),7.25(s,1H),7.24(d,1H),6.85(d,1H),4.06-3.90(m,1H),3.86- 3.66(m,10H),3.36(dd,1H),3.10-2.88(m,5H),2.49(q,2H),2.36-2.25(m,1H),2.02-1.91(m,1H),1.75(s,6H). LCMS[M+H]523.31.

[0486]

[0487] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((((trans)-3-aminocyclobutyl)methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 31)

[0488] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (trans-3-(aminomethyl)cyclobutyl)carbamate. 1 H NMR(500MHz,D2O)δ7.93(d,1H),7.39(d,1H),7.29(d,1H),7.28(d,1H),6.87(d,1H),4.06-3.97(m,1H),3.82(s,4H),3.76(s,5H),3. 71(s,1H),3.44(d,2H),3.12-2.95(m,3H),2.94-2.85(m,1H),2.54-2.45(m,2H),2.45-2.36(m,3H),2.00-1.90(m,1H),1.78(s,6H). LCMS[M+H]537.37.

[0489]

[0490] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((cis-3-aminocyclobutyl)methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 32)

[0491] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (cis-3-(aminomethyl)cyclobutyl)carbamate. 1H NMR(500MHz,D2O)δ7.94(d,1H),7.39(d,1H),7.29(d,1H),7.28(d,1H),6.87(d,1H),3.89-3.72(m,9H),3.72-3.63(m,1H),3. 43-3.31(m,3H),3.10-2.94(m,3H),2.73-2.56(m,3H),2.43-2.32(m,1H),2.13-2.03(m,2H),2.00-1.88(m,1H),1.78(s,6H). LCMS[M+H]537.3.

[0492]

[0493] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3R)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (first eluting diastereomer, compound 33)

[0494] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1R,3S)-3-aminocyclopentyl)carbamate followed by separation by chiral HPLC and deprotection of the optically pure stereoisomers. 1 H NMR(500MHz,D2O)δ8.04(d,1H),7.40(d,1H),7.30(s,1H),7.29(d,1H),6.85(d,1H),4.08-3.99(m,1H),3.86-3.74(m,10H) ,3.42(dd,1H),3.11-3.03(m,3H),2.85-2.76(m,1H),2.43-2.26(m,3H),2.05-1.92(m,3H),1.91-1.83(m,1H),1.78(s,6H). LCMS[M+H]537.4.

[0495]

[0496] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3R)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (second eluting diastereomer, compound 34)

[0497] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1R,3S)-3-aminocyclopentyl)carbamate followed by separation by chiral HPLC and deprotection of the optically pure stereoisomers. 1 H NMR(500MHz,D2O)δ8.04(d,1H),7.40(d,1H),7.30(s,1H),7.29(d,1H),6.85(d,1H),4.08-3.99(m,1H),3.91-3.71(m,10H) ,3.42(dd,1H),3.12-2.97(m,3H),2.86-2.74(m,1H),2.44-2.24(m,3H),2.08-1.92(m,3H),1.92-1.83(m,1H),1.78(s,6H). LCMS[M+H]537.4.

[0498]

[0499] 4-(2-amino-2-methylpropanoyl)-N-(1-(2-(((1S,3S)-3-aminocyclopentyl)amino)-2,3-dihydro-1H-inden-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride

[0500] (Compound 36)

[0501] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(2-oxo-2,3-dihydro-1H-inden-5-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate. 1HNMR(500MHz,D2O)δ7.94(d,1H),7.52(d,1H),7.41(s,1H),7.34(d,1H),6.88(d,1H),4.36-4.20(m,1H),4.13-4.01(m,1H),3.99-3.92 (m,1H),3.82(br.s,4H),3.75(br.s,4H),3.57(dd,2H),3.26(dd,2H),2.52-2.39(m,2H),2.35(t,2H),1.95-1.79(m,2H),1.78(s,6H). LCMS[M+H]523.3.

[0502]

[0503] 4-(2-amino-2-methylpropanoyl)-N-(1-(2-(((trans-3-aminocyclobutyl)methyl)amino)-2,3-dihydro-1H-inden-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 37)

[0504] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(2-oxo-2,3-dihydro-1H-inden-5-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (trans-3-(aminomethyl)cyclobutyl)carbamate. 1 H NMR(400MHz,D2O)δ7.97(d,1H),7.49(d,1H),7.38(s,1H),7.31(d,1H),6.84(d,1H),4.24(t,1H),4.02-3.92(m,1H),3.79(br.s,3H ),3.74(br.s,5H),3.55(dd,2H),3.36(d,2H),3.23(dd,2H),2.92-2.75(m,1H),2.52-2.41(m,2H),2.41-2.31(m,2H),1.75(s,6H). LCMS[M+H]523.3.

[0505]

[0506] 4-(2-amino-2-methylpropanoyl)-N-(1-(2-((trans-4-aminocyclohexyl)amino)-2,3-dihydro-1H-inden-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 38)

[0507] Prepared as in Scheme 3 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(2-oxo-2,3-dihydro-1H-inden-5-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (trans-4-aminocyclohexyl)carbamate. 1 H NMR(400MHz,D2O)δ7.92(d,1H),7.49(d,1H),7.38(s,1H),7.31(d,1H),6.86(d,1H),4.46-4.35(m,1H),3.79(br.s ,3H),3.74(br.s,5H),3.54(dd,2H),3.34(d,2H),3.21(dd,2H),2.41-2.16(m,4H),1.75(s,6H),1.68-1.53(m,4H). LCMS[M+H]537.3.

[0508]

[0509] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)(methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 15)

[0510] Solution 4

[0511]

[0512] Reagents: Step 1) Molecular sieves, 37% aqueous formaldehyde, NaCNBH3, MeOH, room temperature, 16 hours 2) 2M HCl in MeOH

[0513] Step 1: tert-Butyl ((1r,4r)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-b1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(methyl)amino)cyclohexyl)carbamate. Tert-butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-b1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)amino)cyclohexyl)carbamate (0.04 g, 0.05 mmol) and A stirred suspension of molecular sieves (1.5 g) in methanol was added with formaldehyde (30% aqueous solution, 1.5 mL) followed by sodium cyanoborohydride (5 mg, 0.08 mmol) and stirred at room temperature for 16 hours. The reaction mixture was filtered and the filtrate was evaporated. Saturated NaHCO3 aqueous solution was added and the compound was extracted with ethyl acetate (2x10 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure, and the crude material was purified by flash column chromatography to give the title compound (32 mg, 78% yield) as a white solid.

[0514] Step 2: 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. The compound tert-butyl ((1r,4r)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(methyl)amino)cyclohexyl)carbamate (0.03 g, 0.04 mmol) was dissolved in 2M HCl in methanol and stirred at room temperature for 4 hours. The solvent was evaporated and the crude material was purified by HPLC to give the title compound (15 mg, 60% yield) as a brown solid. 1 H NMR(500MHz,D2O): δ8.00(d,1H),7.40–7.36(m,1H),7.27(s,1H),7.26(s,1H),6.82(d,1H),3.98–3.91(m,1H),3.80–3.73(m,8H),3.68 –3.61(m,1H),3.33–2.99(m,5H),2.90(s,3H),2.41–2.26(m,5H),2.09–1.92(m,1H),1.81–1.74(m,1H),1.73(s,6H),1.72–1.58(m,3H). LCMS[M+H]565.4.

[0515]

[0516] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-((6-aminospiro[3.3]hept-2-yl)(methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 11)

[0517] Prepared as in Scheme 4 from tert-butyl (1-(4-((1-(6-((6-((tert-butoxycarbonyl)amino)spiro[3.3]hept-2-yl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate and formaldehyde. LCMS [M+H] 577.2.

[0518]

[0519] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,4S)-4-aminocyclohexyl)(methyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 19)

[0520] Prepared as in Scheme 4 from tert-butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate and formaldehyde. 1 HNMR(500MHz,D2O)δ:7.99(d,1H),7.38–7.34(m,1H),7.25(s,2H),6.80(d ,1H),4.22–4.15(m,1H),3.99–3.87(m,2H),3.83–3.65(m,8H),3.28–3.16 (m,2H),3.15–2.92(m,2H),2.88(s,3H),2.45–2.29(m,4H),2.26–2.18(m, 1H),2.17–2.00(m,1H),1.99–1.84(m,1H),1.83–1.71(m,1H),1.72(s,6H). LCMS[M+H]551.4.

[0521]

[0522] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)(ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 16)

[0523] Solution 5

[0524]

[0525] Reagents: Step 1) Acetaldehyde, NaCNBH3, MeOH, RT, 16 h 2) 2M HCl in MeOH.

[0526] Step 1: tert-Butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(ethyl)amino)cyclohexyl)carbamate. To a stirred solution of tert-butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)amino)cyclohexyl)carbamate (0.04 g, 0.05 mmol) in methanol was added acetaldehyde (0.5 mL) under N2 atmosphere followed by sodium cyanoborohydride (5 mg, 0.08 mmol) and stirred at room temperature for 16 hours. The reaction mixture was filtered and the filtrate was evaporated. Saturated NaHCO3 aqueous solution was added to the residue, and the compound was extracted with ethyl acetate (2x10mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure, and the crude material was purified by flash column chromatography to obtain the title compound (31 mg, 75% yield) as a white solid.

[0527] Step 2: 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)(ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. Tert-butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(ethyl)amino)cyclohexyl)carbamate (0.03 g, 0.04 mmol) was dissolved in 2M HCl in methanol and stirred at room temperature for 4 hours. The solvent was evaporated and the crude material was purified by HPLC to afford the title compound as a brown solid (18 mg, 65% yield). 1H NMR (500MHz, D2O): δ8.04(d,1H),7.37(d,1H),7.27(s,1H),7.26(s,1H),6.82(d,1H),3.98–3.91(m,1H),3.81–3.73(m,8H),3.66–3.59(m, 1H),3.44(q,2H),3.29–2.99(m,5H),2.41–2.26(m,5H),2.09–1.96(m ,1H),1.81–1.76(m,1H),1.75(s,6H),1.72–1.58(m,3H),1.43(t,3H). LCMS[M+H]579.5.

[0528]

[0529] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)(ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 20)

[0530] Prepared as in Scheme 5 from tert-butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate and acetaldehyde. 1 H NMR(500MHz,D2O): δ7.93(d,1H),7.35(d,1H),7.24(s,2H),6.81(d,1H),4.26–4.19(m,1H),3.99–3.87(m,2H),3.83–3.65(m,8H),3.60 –3.45(m,1H),3.33–3.22(m,3H),3.12–2.93(m,2H),2.51–2.29(m,5H),2.10–1.85(m,2H),1.83–1.71(m,1H),1.72(s,6H),1.43(t,3H). LCMS[M+H]565.4.

[0531]

[0532] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)(cyclopropylmethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 17)

[0533] Solution 6

[0534]

[0535] Reagents: Step 1) Cyclopropanecarboxaldehyde, NaCNBH3, MeOH, rt, 16 h 2) 2M HCl in MeOH.

[0536] Step 1: tert-Butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(cyclopropylmethyl)amino)cyclohexyl)carbamate. To a stirred solution of tert-butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)amino)cyclohexyl)carbamate (0.04 g, 0.05 mmol) in methanol under N2 atmosphere was added cyclopropanecarboxaldehyde (0.05 mL) followed by sodium cyanoborohydride (5 mg, 0.08 mmol) and stirred at room temperature for 16 hours. The reaction mixture was filtered and the filtrate was evaporated. Saturated NaHCO3 aqueous solution was added to the residue, and the compound was extracted with ethyl acetate (2x10mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure, and the crude material was purified by flash column chromatography to give the title compound (29 mg, 70% yield) as a white solid.

[0537] Step 2: 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((trans)-4-aminocyclohexyl)(cyclopropylmethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. Tert-butyl ((trans)-4-((6-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)(cyclopropylmethyl)amino)cyclohexyl)carbamate (0.03 g, 0.04 mmol) was dissolved in 2M HCl in methanol and stirred at room temperature for 4 hours. The solvent was evaporated and the crude material was purified by HPLC to give the title compound (17 mg, 70% yield) as a brown solid. 1 H NMR(500MHz,D2O): δ7.98(d,1H),7.35(d,1H),7.25(s,1H),7.24(s,1H),6.80( d,1H),4.19–3.98(m,1H),3.81–3.73(m,8H),3.66–3.61(m,1H),3.33–3.19(m, 5H),3.17–2.96(m,2H),2.40–2.23(m,5H),2.05–1.98(m,1H),1.85–1.78(m,2H ),1.72(s,6H),1.68–1.56(m,2H),1.18–1.12(m,1H),0.77(d,2H),0.43(d,2H). LCMS [M+H] 605.5.

[0538]

[0539] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)(cyclopropylmethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 21)

[0540] Prepared as in Scheme 6 from tert-butyl (1-(4-((1-(6-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate and cyclopropanecarbaldehyde. 1H NMR(500MHz,D2O): δ7.97(d,1H),7.37–7.35(m,1H),7.24(s,1H),7.23(s,1H),6 .80(d,1H),4.27–4.19(m,1H),4.06–3.95(m,1H),3.94–3.86(m,1H),3.83–3.63( m,8H),3.38–3.17(m,3H),3.12–2.93(m,3H),2.51–2.29(m,5H),2.06–1.91(m,2 H),1.83–1.71(m,1H),1.72(s,6H),1.22–1.15(m,1H),0.78(t,2H),0.42(t,2H). LCMS[M+H]591.5.

[0541]

[0542] Exo-4-(2-amino-2-methylpropanoyl)-N-(1-(3-(-6-(aminomethyl)-3-azabicyclo[3.1.0]hex-3-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 12)

[0543] Solution 7

[0544]

[0545] Reagents: Step 1) tert-butyl exo-((3-azabicyclo[3.1.0]hexan-6-yl)methyl)carbamate, NaCNBH3, DCE, rt, 16 h 2) B2Pin2, KOAc, PdCl2(dppf), dioxane, 80°C, 16 h 3) Cytosine, TMEDA, Cu(OAc)2H2O, MeOH:H2O (4:1), air, rt, 20 h (4) 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide, CH3CN, 80°C, 16 h (5) 4M HCl in dioxane, rt, 8 h.

[0546] Step 1: exo-tert-butyl ((3-(7-bromochroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate. To a stirred solution of 7-bromochroman-3-one (0.1 g, 0.5 mmol) in DCE (5 mL) was added exo-tert-butyl ((3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate (0.09 g, 0.5 mmol) and activated Molecular sieve (0.05g). The reaction mixture was stirred at room temperature for 2 hours. NaCNBH3 (0.04g, 0.7mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered and the filtrate was poured into water (30mL), and extracted with DCE (3x15mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure, and purified by flash chromatography (20% EtOAc in hexane) to give the title compound (0.1g, 54%) as a yellow solid. 1 H NMR(400MHz,MeOD)δ6.96-6.94(m,2H),6.88(s,1H),4.24(d,1H),3.08(t,1H),3.13(t,1H),2.93-2.86(m,3H) ,2.68-2.62(m,2H),2.55-2.52(m,2H),1.46(s,9H),1.43-1.33(m,2H),1.30-1.24(m,1H),1.19-1.16(m,1H). LCMS[M+H]423.2.

[0547] Step 2: tert-Butyl exo-3-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate. To a stirred solution of tert-butyl exo-((3-(7-bromochroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate (0.1 g, 0.3 mmol) in dioxane (150 mL) was added B2Pin2 (0.12 g, 0.5 mmol) and KOAc (0.07 g, 0.7 mmol) at room temperature under N2 atmosphere. The reaction mixture was degassed with N2 for 20 min. PdCl2(dppf) (0.02 g, 0.02 mmol) was added and heated at 80°C for 16 h. The reaction mixture was poured into water (30 mL) and extracted with (3 x 15 mL) EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (0.2 g, quantitative) as a brown viscous substance. LCMS [M+H] 471.2.

[0548] Step 3: tert-Butyl ((exo)-3-(7-(4-amino-2-oxopyrimidin-1(2H)-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate. A solution of tert-butyl exo-((3-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate (0.2 g, 0.4 mmol) and cytosine (0.05 g, 0.4 mmol) in MeOH:HO (6 mL, 4:1) was stirred at room temperature for 30 minutes exposed to air. TMEDA (0.07mL, 0.5mmol) and Cu (OAc) 2.HO (0.08g, 0.4mmol) were added, and the reaction mixture was exposed to air stirring at room temperature for 20 hours. The reaction mixture was concentrated under reduced pressure. The crude material was poured into water (30mL) and extracted with EtOAc (3x15mL). The combined organic phases were dried over Na2SO4, filtered, and concentrated under reduced pressure, and purified by flash chromatography (10% MeOH in DCM) to give the title compound (0.045g, 24%) as a brown solid. 1 HNMR: (400MHz, DMSO-d6): 7.53(d,1H),7.19-7.07(m,2H),6.76-6.70(m,2H),6.67(d,1H),5.80-5.72(m,1H),4.21(d,1H),3.79(t,1H),3.06-2.99(m,2 H),2.90(d,2H),2.76-2.73(m,2H),2.66-2.63(m,2H),1.35(s,9H),1.30-1.28(m,3H),1.27-1.22(m,1H),1.06-1.04(m,1H). LCMS[M+H]454.9.

[0549] Step 4: tert-butyl exo-((3-(7-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hexan-6-yl)methyl)carbamate. To a stirred solution of tert-butyl ((exo)-3-(7-(4-amino-2-oxopyrimidin-1(2H)-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate (0.05 g, 0.1 mmol) and 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide (0.08 g, 0.2 mmol) in acetonitrile (2 mL) was heated at 80° C. for 16 h. The reaction mixture was concentrated under reduced pressure and the crude material was purified by flash chromatography (2% MeOH in DCM) to give the title compound (0.08 g) as a brown sticky material. LCMS [M+H] 751.

[0550] Step 5: Exo-4-(2-amino-2-methylpropanoyl)-N-(1-(3-(-6-(aminomethyl)-3-azabicyclo[3.1.0]hex-3-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. To a stirred solution of exo-tert-butyl((3-(7-(4-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)chroman-3-yl)-3-azabicyclo[3.1.0]hex-6-yl)methyl)carbamate (0.08 g, 0.1 mmol) in dioxane (1.0 mL) was added 4M HCl in dioxane (0.5 mL). The reaction mixture was stirred at room temperature for 8 hours. The reaction mixture was concentrated under reduced pressure and purified by preparative HPLC purification to give the title compound (0.005 g, 8%) as a white solid. 1 H NMR: (400Mz,D2O): δ7.91(d,1H),7.42(d,1H),7.13(q,1H),7.07(d,1H),6.91(d,1H),4.60-4.55(m,1H),4.55-4.48(m,1H),4.13-4. 04(m,2H),3.84-3.79(m,8H),3.68-3.49(m,2H),3.35-3.20(m,2H),3.03(d,2H),2.21-2.04(m,2H),1.80(s,6H),1.68-1.55(m,1H). LCMS[M+H]551.3.

[0551]

[0552] Exo-4-(2-amino-2-methylpropanoyl)-N-(1-(3-(-6-amino-3-azabicyclo[3.1.0]hex-3-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 13)

[0553] Prepared in a manner analogous to Scheme 7 from 7-bromochroman-3-one and exo-(3-azabicyclo[3.1.0]hexan-6-yl)carbamic acid tert-butyl ester. 1 H NMR: (400MHz, D2O): δ7.71(d,1H),7.20(d,1H),6.91(d,1H),6.85(s,1H),6.69(d,1H),4.37(d,1H),4.21(d ,1H),3.90-3.84(m,2H),3.62-3.50(m,9H),3.41-3.31(m,3H),3.04-2.91(m,2H),2.32(s,2H),1.58(s,6H). LCMS[M+H]537.3.

[0554]

[0555] 4-(2-amino-2-methylpropanoyl)-N-(1-(3-((S)-3-(aminomethyl)pyrrolidin-1-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-carboxamide hydrochloride (Compound 14)

[0556] Prepared in an analogous manner to Scheme 7 from 7-bromochroman-3-one and (R)-tert-butyl (pyrrolidin-3-ylmethyl)carbamate. 1 H NMR: (400MHz, D2O): δ7.70(d,1H),7.21(d,1H),6.91(d,1H),6.86(s,1H),6.69(d,1H),4.47(d,2H),4.24(t,2H),3.87(bs,1H), 3.69-3.55(m,7H),3.45-3.34(m,2H),3.25-3.18(m,1H),3.12-3.01(m,4H),2.65-2.52(m,2H),2.40-2.26(m,2H),1.57(s,6H). LCMS[M+2H] / 2 270.2.

[0557]

[0558] 4-(2-amino-2-methylpropanoyl)-N-(1-(3-((R)-3-(aminomethyl)pyrrolidin-1-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 18)

[0559] Prepared in an analogous manner to Scheme 7 from 7-bromochroman-3-one and (S)-tert-butyl (pyrrolidin-3-ylmethyl)carbamate. 1 H NMR: (400MHz, D2O): δ7.72(d,1H),7.21(d,1H),6.91(d,1H),6.85(s,1H),6.68(d,1H),4.46(d,2H),4.23 (t,2H),3.86(bs,2H),3.61-3.50(m,8H),3.07(d,2H),3.15-2.95(m,4H),2.41-2.26(m,2H),1.56(s,6H). LCMS[M+H]539.8.

[0560]

[0561] 4-(2-amino-2-methylpropanoyl)-N-(1-(3-(4-aminoazepan-1-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 22)

[0562] Prepared in a similar manner to Scheme 7 from 7-bromochroman-3-one and tert-butyl azepan-4-ylcarbamate. 1 H NMR: (400MHz, D2O): δ7.76(d,1H),7.25(d,1H),6.95(d,1H),6.89(s,1H),6.73-6.71(m,1H),4.56(d,2H),4.32(t,1H),3.96(bs ,1H),3.85-3.62(m,4H),3.60-3.29(m,9H),3.28-3.16(m,2H),2.85(d,1H),2.35-2.22(m,2H),2.15-1.90(m,2H),1.61(s,6H). LCMS[M+H]553.5.

[0563]

[0564] 4-(2-amino-2-methylpropanoyl)-N-(1-(3-(1-(aminomethyl)-3-azabicyclo[3.1.0]hex-3-yl)chroman-7-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 23)

[0565] Prepared in a manner analogous to Scheme 7 from 7-bromochroman-3-one and tert-butyl ((3-azabicyclo[3.1.0]hex-1-yl)methyl)carbamate. 1 H NMR: (400MHz,D2O):7.76(d,1H),7.19(d,1H),6.90(d,1H),6.84(s,1H),6.66(d,1H),4.32-422(m,3H),3.86(b s,2H),3.78-3.47(m,11H),3.35-3.31(m,3H),3.12-2.80(m,3H),1.92(bs,1H),1.56(s,6H),1.21-0.95(m,2H). LCMS[M+H]551.3.

[0566]

[0567] 4-(2-amino-2-methylpropanoyl)-N-(1-(2-(4-aminocyclohexyl)-1,2,3,4-tetrahydroisoquinolin-6-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 35) Scheme 8

[0568]

[0569] Reagents: 1) tert-butyl (4-oxocyclohexyl)carbamate, Na(OAc)3BH, DCE, room temperature for 16 hours 2) Pd(dppf)Cl2·CH2Cl2, KOAc, B2Pin2, 1,4-dioxane, 105°C, 16 hours 3) cytosine, Cu(OAc)2·H2O, TMEDA, MeOH:H2O (4:1), room temperature for 48 hours 4) 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide, MeCN, 85°C, 16 hours 5) HCl, MeOH, room temperature for 4 hours.

[0570] Step 1: tert-Butyl (4-(6-bromo-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate. To a stirred solution of tert-butyl (4-oxocyclohexyl)carbamate (251 mg, 1.2 mmol) in DCE (10 mL) was added 6-bromo-1,2,3,4-tetrahydroisoquinoline (250 mg, 1.2 mmol) and Na(OAc)3BH (375 mg, 1.8 mmol). The reaction was stirred at room temperature for 16 hours. The reaction mixture was diluted with CH2Cl2 and washed with saturated NaHCO3 (1x15 mL). The aqueous layer was extracted with CH2Cl2 (2x20 mL), and the combined organics were dried over Na2SO4, concentrated under reduced pressure and purified by column chromatography (CH2Cl2:MeOH) to give the desired product. LCMS [M+H] 411.1.

[0571] Step 2 Tert-butyl (4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate. A flask containing tert-butyl (4-(6-bromo-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate (512 mg, 1.25 mmol), Pd(dppf)Cl2.CH2Cl2 (30 mg, 0.03 mmol), KOAc (368 mg, 3.75 mmol) and B2pin2 (349 mg, 1.38 mmol) was evacuated and flushed with N2. 1,4-Dioxane (10 mL) was added, and the reaction mixture was degassed and heated to 105 °C for 16 h. The reaction mixture was cooled to room temperature, diluted with EtOAc (30 mL), and filtered through The crude reaction mixture was purified by flash chromatography (EtOAc:Hex) to give the desired product. LCMS [M+H] 457.8.

[0572] Step 3: tert-Butyl (4-(6-(4-amino-2-oxopyrimidin-1(2H)-yl)-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate. A suspension of tert-butyl (4-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate (500 mg, 1.1 mmol) and cytosine (122 mg, 1.1 mmol) in MeOH:H2O (4:1, 100 mL) was stirred at room temperature for 30 minutes in an air atmosphere. Cu(OAc)2·H2O (219 mg, 1.1 mmol) and TMEDA (0.20 mL, 1.3 mmol) were added and the reaction was stirred at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure and H2O (150 mL) was added. The solid was collected by filtration and washed with Et2O (25 mL) and cold H2O (25 mL) to give the desired product. LCMS [M+H] 440.3.

[0573] Step 4: tert-butyl (1-(4-((1-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)-1,2,3,4-tetrahydroisoquinolin-6-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. To a solution of tert-butyl (4-(6-(4-amino-2-oxopyrimidin-1(2H)-yl)-3,4-dihydroisoquinolin-2(1H)-yl)cyclohexyl)carbamate (124 mg, 0.28 mmol) in MeCN (20 mL) was added 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide (200 mg, 0.39 mmol). The reaction mixture was heated to 85 °C for 16 hours. The volatiles were removed under reduced pressure, and the crude solid was dissolved in CH2Cl2 (100 mL), washed with H2O (1x100 mL), and concentrated under reduced pressure. The crude solid was purified by flash chromatography (CH2Cl2:MeOH:NH4OH) to give the desired product.

[0574] Step 5: 4-(2-amino-2-methylpropanoyl)-N-(1-(2-(4-aminocyclohexyl)-1,2,3,4-tetrahydroisoquinolin-6-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. Tert-butyl (1-(4-((1-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)-1,2,3,4-tetrahydroisoquinolin-6-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate was dissolved in a solution of HCl / MeOH (5 mL) and stirred for 4 hours. The volatiles were removed under reduced pressure and the crude solid was purified by reverse phase HPLC (H2O:CH3CN:TFA) and concentrated under reduced pressure. Addition of HCl / MeOH (3 x 15 mL) and evaporation under reduced pressure gave the desired product. 1 HNMR(500MHz,D2O)δ8.05(d,1H),7.49-7.37(m,3H),6.87(dd,1H),4.64(d,2H),3.99-3.49(m,10H),3.44-3 .23(m,3H),2.47-2.28(m,2H),2.26-2.11(m,2H),2.03(d,2H),1.87(d,1H),1.78(s,6H),1.73-1.59(m,1H). LCMS[M+H]537.4.

[0575]

[0576] 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(4-(2-aminoethyl)piperidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 42)

[0577] Solution 9

[0578]

[0579] Reagents: 1) DMP, CH2Cl2, room temperature, 2 hours 8) NaCNBH3, DCM, 45°C, 16 hours 2) 4M HCl / dioxane, room temperature, 3 hours

[0580] Step 8: tert-butyl (1-(4-((1-(6-(4-(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. To a stirred solution of tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate (0.25 g, 0.45 mmol) and tert-butyl (2-(piperidin-4-yl)ethyl)carbamate (0.15 g, 0.67 mmol) in DCM (10 mL) was added NaBH3CN (0.06 g, 0.9 mmol) at room temperature under N2 atmosphere. The reaction mixture was stirred at 40 °C for 16 hours. The reaction mixture was concentrated under reduced pressure and purified by column chromatography (12% MeOH in DCM) to give the title compound (0.1 g, 29%) as an off-white solid. LCMS [M+H] 765.6.

[0581] Step 9: 4-(2-amino-2-methylpropanoyl)-N-(l-(6-(4-(2-aminoethyl)piperidin-l-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)piperazine-l-carboxamide hydrochloride. To a stirred solution of tert-butyl (l-(4-((l-(6-(4-(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-l-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-l-yl)-2-methyl-l-oxopropan-2-yl)carbamate (0.1 g, 0.13 mmol) in dioxane (2 mL) was added 4M HCl in dioxane (2 mL). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure and purified by HPLC to give the title compound (0.025 g, 33%) as a pale yellow solid. 1 H NMR: (400MHz, D2O): δ7.79(d,1H),7.20(d,1H),7.08(d,2H),6.66(d,1H),3.61-3.49(m,11H),3.18-3.14(m, 1H),3.07-2.81(m,7H),2.19(bs,1H),1.94(d,2H),1.83-1.78(m,1H),1.63-1.51(m,9H),1.43-1.34(m,2H). LCMS[M / 2+H]283.3.

[0582]

[0583] 4-(2-amino-2-methylpropanoyl)-N-(1-(7-(4-aminoazepan-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 43)

[0584] Prepared in a manner similar to Scheme 9 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl azepan-4-ylcarbamate. 1 H NMR: (400MHz, D2O): LCMS [M / 2+H] 276.3.

[0585]

[0586] 4-(2-amino-2-methylpropanoyl)-N-(1-(7-(3-aminoazetidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 39)

[0587] Prepared in a manner similar to Scheme 9 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl azetidin-3-ylcarbamate. 1 H NMR: (400MHz, D2O): δ7.83(d,1H),7.24(d,1H),7.14(d,2H),6.71(d,1H),4.70-4.60(m,2H),4.41(s,3H),3.79( bs,2H),3.66-3.54(m,7H),3.24-3.20(m,1H),2.87-2.75(m,3H),2.17(bs,1H),1.78-1.72(m,1H),1.62(s,6H). LCMS[M+H]509.5.

[0588]

[0589] 4-(2-amino-2-methylpropanoyl)-N-(1-(7-(3-(aminomethyl)azetidin-1-yl)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 40)

[0590] Prepared in a manner analogous to Scheme 9 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (azetidin-3-ylmethyl)carbamate. 1 H NMR: (400MHz, D2O): δ7.82(d,1H),7.23(d,1H),7.14(d,2H),6.71(d,1H),4.36(t,2H),4.12-4.07(m,2H),3.66-3. 62(m,8H),3.34(d,1H),3.23-3.15(m,4H),2.86(d,2H),2.76-2.72(m,1H),2.15(bs,1H),1.70(m,1H),1.62(s,6H). LCMS[M+H]523.4.

[0591]

[0592] trans-4-(2-amino-2-methylpropanoyl)-N-(1-(7-((3-(aminomethyl)cyclobutyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 41)

[0593] Prepared in a manner analogous to Scheme 9 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (((trans)-3-aminocyclobutyl)methyl)carbamate. 1H NMR: (400MHz, D2O): δ7.77(d,1H),7.22(d,1H),7.12(s,2H),6.72(d,1H),4.07(d,1H),3.66-3.61(m,9H),3.20-3.10 (m,3H),2.91-2.70(m,3H),2.70-2.66(m,1H),2.50-2.44(m,2H),2.26-2.17(m,3H),1.90-1.81(m,1H),1.62(s,6H). LCMS[M+H]537.4.

[0594]

[0595] cis-4-(2-amino-2-methylpropanoyl)-N-(1-(7-((3-(aminomethyl)cyclobutyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 44)

[0596] Prepared in a manner analogous to Scheme 9 from tert-butyl (2-methyl-1-oxo-1-(4-((2-oxo-1-(6-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)propan-2-yl)carbamate and tert-butyl (((cis)-3-aminocyclobutyl)methyl)carbamate. 1 H NMR: (400MHz, D2O): δ7.75(d,1H),7.22(d,1H),7.12(s,2H),6.73(d,1H),3.91(bs,1H),3.67-3.61(m,8H),3.31-3.1 5(m,3H),3.03-2.88(m,4H),2.57-2.45(m,3H),2.31-2.18(m,1H),2.09-1.96(m,2H),1.90-1.83(m,1H),1.63(s,6H). LCMS[M / 2+H]269.3.

[0597]

[0598] 4-(2-amino-2-methylpropanoyl)-N-(1-(2-((trans)-4-aminocyclohexyl)isoindolin-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride (Compound 46)

[0599] Solution 10

[0600]

[0601] Reagents: Step 1) (tert-butyl (1r, 4r)-4-aminocyclohexyl)carbamate, K2CO3, CH3CN, 85°C, 16 hours 2) B2Pin2, KOAc, PdCl2(dppf), dioxane, 100°C, 16 hours 3) Cytosine, TMEDA, Cu(OAc)2.H2O, MeOH:H2O (4:1), air, room temperature, 16 hours (4) 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide, CH3CN, 80°C, 16 hours (5) AcCl, MeOH, room temperature, 4 hours.

[0602] Step 1: tert-butyl ((trans)-4-(5-bromoisoindolin-2-yl)cyclohexyl)carbamate. To a stirred solution of 4-bromo-1,2-bis(bromomethyl)benzene (0.31 g, 0.9 mmol) in CH3CN (15 mL) was added tert-butyl ((trans)-4-aminocyclohexyl)carbamate (0.21 g, 1.0 mmol) and K2CO3 (0.27 g, 2.0 mmol) at room temperature. The reaction mixture was stirred at 85 °C for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (20% EtOAc in hexanes) to give the title compound (0.2 g, 56%) as a light yellow solid.

[0603] Step 2: tert-butyl ((trans)-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)cyclohexyl)carbamate. To a stirred solution of tert-butyl ((trans)-4-(5-bromoisoindolin-2-yl)cyclohexyl)carbamate (0.1 g, 0.25 mmol) in dioxane (5 mL) was added B2Pin2 (0.08 g, 0.3 mmol) and KOAc (0.07 g, 0.8 mmol) at room temperature under N2. The reaction mixture was degassed with N2 for 20 min and PdCl2(dppf) (6.0 mg, 0.03 mmol) was added and the reaction mixture was stirred at 100°C for 16 h. The reaction mixture was filtered through celite and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (75% EtOAc in hexanes) to give the title compound (0.08 g, 73%) as a brown solid.

[0604] Step 3: tert-Butyl ((trans)-4-(5-(4-amino-2-oxopyrimidin-1(2H)-yl)isoindolin-2-yl)cyclohexyl)carbamate. A solution of tert-butyl ((trans)-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)cyclohexyl)carbamate (80.0 mg, 0.18 mmol) and cytosine (20 mg, 0.18 mmol) in MeOH:H2O (6 mL, 4:1) was stirred for 30 minutes. TMEDA (25.0 L, 0.22mmol) and Cu(OAc)2.HO (36.0mg, 0.18mmol), and the reaction mixture was exposed to air and stirred for 16 hours at room temperature. The reaction mixture was concentrated under reduced pressure. The crude material was poured into water (30mL) and extracted with CHCl3 (3x15mL). The combined organic phases were dried over Na2SO4, filtered, and concentrated under reduced pressure, and purified by flash chromatography (8:2:0.2MeOH:DCM:NH4OH) to give the title compound (32mg, 44%) as a white solid.

[0605] Step 4: tert-butyl (1-(4-((1-(2-((trans)-4-((tert-butoxycarbonyl)amino)cyclohexyl)isoindolin-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamate. To a stirred solution of tert-butyl ((trans)-4-(5-(4-amino-2-oxopyrimidin-1(2H)-yl)isoindolin-2-yl)cyclohexyl)carbamate (32 mg, 0.08 mmol) in acetonitrile (4 mL) was added 1-(4-(2-((tert-butoxycarbonyl)amino)-2-methylpropanoyl)piperazine-1-carbonyl)-3-methyl-1H-imidazol-3-ium iodide (80 mg, 0.15 mmol), and the reaction mixture was stirred at 80° C. under N2 for 16 h. The reaction mixture was concentrated under reduced pressure, and the crude material was purified by flash chromatography (DCM / MeOH) to give the title compound (35 mg, 64%) as a pale yellow solid.

[0606] Step 5: 4-(2-amino-2-methylpropanoyl)-N-(1-(2-((trans)-4-aminocyclohexyl)isoindolin-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide hydrochloride. Compound (1-(4-((1-(2-((trans)-4-((tert-butoxycarbonyl)amino)cyclohexyl)isoindolin-5-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)piperazin-1-yl)-2-methyl-1-oxopropan-2-yl)carbamic acid tert-butyl ester (30 mg, 0.04 mmol) was dissolved in freshly prepared methanolic HCl (4 mL) and stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure and purified by RPHPLC. The desired fractions were collected and evaporated under reduced pressure. To the residue was added HCl in MeOH (2 x 5 mL) and the solvent was evaporated to give the title compound (11 mg, 50%) as a white solid. 1 H NMR: (400Mz,D2O): δ8.03(d,1H),7.61(d,1H),7.55–7.48(m,2H),6.85(d,1H),4.98(d,2H),4.73(d,3H),3.80 –3.73(m,8H),3.67–3.59(m,1H),3.36–3.26(m,1H),2.42(d,2H),2.27(d,2H),1.75(s,6H),1.74–1.56(m,4H). LCMS[M+H]524.3.

[0607]

[0608] Methyl ((1S,3S)-3-((6-(4-(4-(2-amino-2-methylpropionyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)amino)cyclopentyl)carbamate hydrochloride (Compound 45)

[0609] Solution 11

[0610]

[0611] Reagents: Step 1) NaHCO3, methyl chloroformate, THF, H2O, 0°C to room temperature, 2 hours.

[0612] Step 1: Methyl ((1S,3S)-3-((6-(4-(4-(2-amino-2-methylpropanoyl)piperazine-1-carboxamido)-2-oxopyrimidin-1(2H)-yl)-1,2,3,4-tetrahydronaphthalen-2-yl)amino)cyclopentyl)carbamate: 4-(2-amino-2-methylpropanoyl)-N-(1-(6-(((1S,3S)-3-aminocyclopentyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)piperazine-1-carboxamide (160 mg, 0.3 mmol) was dissolved in saturated aqueous NaHCO3 solution (2 ml) and THF (2 ml). Methyl chloroformate (28 mg, 0.3 mmol) was added at 0°C. The reaction mixture was warmed to room temperature and stirred for 2 hours. The solid was filtered and the residue was concentrated under reduced pressure.The residue was dissolved in MeOH (0.5 ml) and 2M HCl in ether (3 ml) was added and the resulting solid was filtered, washed with ether and dried under vacuum to give the title compound (51 mg, 31%) as a white solid. 1H NMR(500MHz,D2O)δ8.10(dd,1H),7.39(d,1H),7.28(dd,2H),6.82(d,1H),4.20–4.12(m,1H),4.06(q,1H),3.79(d,9H),3.68(s,3 H),3.39(d,1H),3.08–2.95(m,3H),2.36(s,2H),2.22(dd,1H),2.14(td,2H),1.98–1.88(m,1H),1.76(d,7H),1.72–1.61(m,1H). LCMS[M+2H / 2]=298.3.

[0613] Biological Examples

[0614] Standard microbial activity:

[0615] Testing was performed using a certified BSL-2 laboratory. Compounds were evaluated using broth microdilution minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays defined by the Clinical and Laboratory Standards Institute (CLSI) in the M26-A guidelines for Staphylococcus aureus (S. aureus) (Sa), Escherichia coli (E. coli) (Ec), Klebsiella pneumoniae (K. pneumoniae) (Kp), Acinetobacter baumannii (A. baumannii) (Ab), Enterococcus faecalis (E. faecalis) (Ef), and Pseudomonas aeruginosa (P. aeruginosa) (Pa).

[0616] E. coli S30 extracts: Inhibition of bacterial protein synthesis was measured using the E. coli S30 circular DNA extraction system (Promega catalog #L-2010) and luciferase assay reagent (Promega catalog #E1500) with slight modifications to published protocols. Fyfe, C., Sutcliffe, JA and Grossman, TH (2012) "Development and characterization of a Pseudomonas aeruginosa in vitro coupled transcription-translation assay system for evaluation of translation inhibitors" J. Microbiol. Methods 90(3), 256–261.

[0617] Compounds were serially diluted in 0.5 mL microcentrifuge tubes by mixing and transferring 50 μL from the highest concentration to 50 μL water, mixing and transferring 50 μL of this 2-fold dilution to 50 μL water. Repeat this mixing and transfer so that there are 8 tubes in total, the serial dilutions of the compounds in the tubes are 10 times of the required screening concentration and are finally diluted to 1 times by adding S30 luciferase synthesis mixture. The serial dilutions of the compounds are added (2 μL) to the holes of black round-bottom 96-well plates. Water (2 μL) is used as a "no inhibitor" control with 4 holes / plates. No DNA control reaction mixture (20 μL; see below) is used as a control with 4 holes / plates to obtain background luminescence. S30 luciferase synthesis mixture (18 μL; see below) is added to the holes containing the compound or water mixture, and incubated at 37 ° C for 1 hour. The reaction is terminated by transferring to 4 ° C refrigerators for 5 minutes, and then 25 μL luciferase activity mixture is added. Luminescence was measured using a BioTek Synergy HTX plate reader. The % inhibition was determined relative to the no inhibitor control.

[0618] S30 Luciferase Synthesis Mix:

[0619] 445 μL S30 extract, circular

[0620] 712μL S30 premix without amino acids

[0621] 4.45 μL pBESTluc TM DNA (1 μg / μL)

[0622] 78μL Complete Amino Acid Mix

[0623] 267 μL water

[0624] No DNA Control:

[0625] 20 μL S30 extract, circular

[0626] 32 μL S30 premix without amino acids

[0627] 7 μL Complete Amino Acid Mix

[0628] 21 μL water

[0629] Rabbit reticulocyte lysate

[0630] The inhibition of eukaryotic protein synthesis was determined using the rabbit reticulocyte lysate system (catalog number #L-4960) from Promega's nuclease treatment, with slight modifications to the manufacturer's protocol. Compounds were serially diluted in 0.5 mL microcentrifuge tubes by mixing and transferring 50 μL of 50 μL of this 2-fold dilution from the highest concentration. Repeat this mixing and transfer so that there are 8 tubes in total, in which the serial dilutions of the compound are 10 times of the required screening concentration and are finally diluted to 1 times by adding rabbit reticulocyte luciferase synthesis mixture. The serial dilutions of the compound (2.5 μL) are added to the holes of a black round-bottom 96-well plate. Water (2.5 μL) is used as a "no inhibitor" control with 4 holes / plates. No RNA control reaction mixture (2 μL; see below) is used as a control with 4 holes / plates to obtain background luminescence. Rabbit reticulocyte luciferase synthesis mixture (22.5 μL; see below) was added to the wells containing the compound or water mixture and incubated at 30°C for 90 minutes. Luciferase assay reagent (25 μL) was added and luminescence was measured using a BioTek Synergy HTX plate reader. The % inhibition was determined relative to the no inhibitor control.

[0631] Rabbit Reticulocyte Luciferase Synthesis Mix:

[0632] 1,000 μL rabbit reticulocyte lysate

[0633] 5.7 μL luciferase control RNA (1 μg / μL)

[0634] 26μL Complete Amino Acid Mix

[0635] 395 μL water

[0636] No RNA control

[0637] 70 μL rabbit reticulocyte lysate

[0638] 2 μL Complete Amino Acid Mix

[0639] 28 μL water

[0640] Minimum inhibitory concentration (MIC)

[0641] MICs were determined using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method with slight modifications. Clinical and Laboratory Standards Institute (2012). "Methods for dilutionantimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, 9th edition M07-A9. Clinical and Laboratory Standards Institute, Wayne, PA." Serial two-fold dilutions of compounds were prepared in sterile clear round-bottom 96-well plates.

[0642] To prepare microdilution trays, two-fold dilutions of the antimicrobial agent were prepared in the following growth media: cation adjusted Miller-Hinton broth (CAMHB) or CAMHB supplemented with sodium bicarbonate (6.25 or 25 mM final concentration, prepared from 1.0 M stock solution) or CAMHB supplemented with heat inactivated human serum (Fisher catalog # BP2657100) 0%-50%, the preparation being performed by adding 200 μL of the highest concentration to be tested (e.g., 64 μg / mL) in row A, mixing and transferring 100 μL in row A to 100 μL of growth medium in row B, and then repeating the mixing and transfer until row H of the 96-well plate, discarding the remaining excess 100 μL. This slight modification of the CLSI protocol allows for the evaluation of the MIC of 3 compounds per plate in triplicate, although there are only 8 compound dilutions (the CLSI protocol allows 2 compounds, in triplicate, 10 dilutions). The cells were cultured by adding 5 μL of a 1:10 dilution of a 0.5 McFarland suspension (1 × 10 8 CFU / mL), and the bacterial suspension was added to 5×10 4 The final concentration of CFU / well was 1.50 μg / mL. The bacterial suspension was prepared using the growth method described by CLSI. A sterile loop was used to select well-separated colonies (3-5 from agar plates) and used to inoculate a tube containing 4 mL of CAMHB. The culture was incubated at 35°C ± 2°C until the turbidity of the 0.5 McFarland standard was reached or exceeded, as measured by A 600nm When growth exceeds the 0.5 McFarland standard, the turbidity is adjusted to the equivalent of the 0.5 McFarland standard with broth.

[0643] The data for the compounds are provided in Table 6. IC of 1 μM or greater 50 Values ​​(μM) (inhibition % ≤ 50% @ 1 μM) are indicated by "+". IC values ​​of 0.5 μM or greater and less than 1 μM 50 Values ​​(inhibition %>50% and ≤90% @ 1 μM) are indicated by "++". IC values ​​less than 0.5 μM 50 Values ​​(inhibition %>90%@1μM) are indicated by "+++". MIC values ​​(μg / mL) of 32μg / mL or more are indicated by "+". MIC values ​​(μg / mL) of 8μg / mL or more and less than 32μg / mL are indicated by "++". MIC values ​​(μg / mL) less than 8μg / mL are indicated by "+++". "NA" indicates not available.

[0644] Table 6. Biological activities of compounds of formula I or their pharmaceutically acceptable salts

[0645]

[0646]

[0647] Table 6. Biological activities of the compounds of formula I or their pharmaceutically acceptable salts

[0648]

[0649]

[0650] For the purpose of clarity and understanding, the foregoing disclosure has been described in detail by way of illustration and example. The present invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many changes and modifications may be made while remaining within the spirit and scope of the present invention. It will be apparent to those skilled in the art that changes and modifications may be implemented within the scope of the appended claims. Therefore, it should be understood that the above description is intended to be illustrative and not restrictive. Therefore, the scope of the present invention should not be determined with reference to the above description, but instead should be determined with reference to the following appended claims together with the full scope of equivalents to which the claims are entitled.

Claims

1. A compound of formula I: or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein: Ring A is a 3-8 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2 and oxo; J is C1-C6 alkylene or C3-C8 cycloalkylene, any of which is optionally substituted with halo, OH or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently replaced with O, S, SO, SO2 or C=O; R x , R y , R x’ and R y’ Each is independently H, C1-C6 alkyl or amino protecting group; Y is a bond or a C1-C6 alkylene optionally substituted with OH, NH2, CN, halo or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently substituted with O, NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl), N-(C1-C6 alkoxy) 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N-(C 1-6 alkyl)(C=O) or (C=O); Ring B is a 3-8 membered monocyclic cycloalkylene, a 3-8 membered monocyclic heterocycloalkylene, a 6-12 membered bicyclic cycloalkylene, or a 6-12 membered bicyclic heterocycloalkylene, each of which is optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl; L is a bond or C1-C6 alkylene, wherein up to two methylene units of the C1-C6 alkylene can be independently replaced by O, NH, (C=O), NH(C=O), N-(C 1-6 alkyl)(C=O), (C=NH), NH(C=N) or N-(C 1-6 Alkyl) substitution; Ring C and the benzene ring to which it is fused together form an 8-12 membered bicyclic arylene or 8-12 membered bicyclic heteroarylene, wherein the bicyclic heteroarylene has 1-3 heteroatoms independently selected from N, O or S, wherein in the 8-12 membered bicyclic arylene or 8-12 membered bicyclic heteroarylene, at least one ring in the bicyclic ring system is aromatic; R 1 , R 2 and R 3 Each is independently selected from the group consisting of C1-C6 alkyl, halo, CN, OH, NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, COO(C1-C6 alkyl), CONH2, C1-C6 haloalkyl, oxo and C1-C6 alkoxy; and m, n and p are each independently 0, 1, 2 or 3.

2. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring A is a 5-6 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2 and oxo.

3. The compound according to claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring A is Each R 4 Independently selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, phenyl, OH, NH2 and oxo, wherein q is 0, 1 or 2.

4. The compound according to claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring A is 5. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein J is a C1-C6 alkylene group optionally substituted with halo, OH or C1-C6 alkoxy, wherein up to two methylene units of the C1-C6 alkylene group are optionally and independently replaced with O, S, SO, SO2 or C=O.

6. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein J is a C1-C6 alkylene group optionally substituted with OH, wherein one methylene unit of the C1-C6 alkylene group is replaced with C=O.

7. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein J is 8. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein R x and R y It's H.

9. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein yes 10. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein yes 11. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Selected from the group consisting of: wherein each X1 is independently selected from CH2, CH, O, S, SO, SO2, N and NH, and wherein R 2 and R 3 Each is independently C1-C6 alkyl, halo or C1-C6 haloalkyl, wherein n and p are each independently 0, 1 or 2.

12. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Selected from the group consisting of:

13. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Selected from the group consisting of:

14. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Y is a bond.

15. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Y is a C1-C3 alkylene optionally substituted with OH, NH2, halo, or C1-C6 alkoxy, and wherein one methylene unit of the C1-C3 alkylene is optionally substituted with O, NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl), N-(C1-C6 alkoxy), 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N-(C 1-6 alkyl)(C═O) or (C═O).

16. The compound of claim 15, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Y is selected from the group consisting of O, NH, NH-C 1-2 Alkylene, N(C1-C6 alkyl), N(C1-C6 hydroxyalkyl), N(C1-C6 haloalkyl), N(C1-C6 1-6 Alkylene-C 3-8 Cycloalkyl), NH(C=O), N(C 1-6 alkyl)(C=O) and (C=O).

17. The compound of claim 16, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Y is selected from the group consisting of NH, N(C1-C6 alkyl), NH-C 1-2 Alkylene and N(C 1-6 Alkylene-C 3-8 cycloalkyl).

18. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Y is selected from the group consisting of -NH-, -NMe-, -NEt-, -NH-CH2-, and -N(CH2-cyclopropyl)-.

19. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Selected from the group consisting of:

20. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 3-8 membered monocyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

21. The compound of claim 20, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 4-6 membered monocyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl.

22. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein B is a 3-8 membered monocyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COOH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2, and C1-C6 hydroxyalkyl.

23. The compound of claim 22, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B is a 4-7 membered monocyclic heterocycloalkylene optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl, and wherein B contains up to 2 nitrogen atoms.

24. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-10 membered bicyclic cycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, C1-C6 haloalkyl, OH and C1-C6 hydroxyalkyl.

25. The compound of claim 24, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-10 membered fused, spiro or bridged bicyclic cycloalkylene.

26. The compound of claim 25, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-10 membered fused bicyclic cycloalkylene group.

27. The compound of claim 25, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-10 membered bridged bicyclic cycloalkylene.

28. The compound of claim 25, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-10 membered spirobicyclocycloalkylene.

29. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-12 membered bicyclic heterocycloalkylene group optionally substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, C1-C6 haloalkyl, OH, and C1-C6 hydroxyalkyl.

30. The compound of claim 29, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-9 membered fused, spiro or bridged bicyclic heterocycloalkylene containing up to 3 nitrogen atoms.

31. The compound of claim 29 or 30, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a 6-9 membered fused bicyclic heterocycloalkylene group containing up to 2 nitrogen atoms.

32. The compound of claim 29 or 30, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a spirobicyclic heterocycloalkylene containing up to 2 nitrogen atoms.

33. The compound of claim 29 or 30, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is a bridged bicyclic heterocycloalkylene containing up to 2 nitrogen atoms.

34. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring B is selected from the group consisting of:

35. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L is a bond.

36. The compound of claim 1, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L is C1-C6 alkylene, wherein up to two methylene units of the C1-C6 alkylene are optionally and independently replaced by O, NH, (C=O), NH(C=O), N-(C 1-6 alkyl)(C=O), (C=NH), NH(C=N) or N-(C 1-6 Alkyl) replacement.

37. The compound of claim 36, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L is a C1-C6 alkylene group.

38. The compound of claim 37, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L is -CH2- or -CH2CH2-.

39. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein R x’ and R y’ Each is independently selected from the group consisting of H, Boc and methoxycarbonyl.

40. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Selected from the group consisting of:

41. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3 Each is independently selected from the group consisting of a C1-C6 alkyl group, a halogen group, a C1-C6 haloalkyl group, an oxo group, and a C1-C6 alkoxy group, and m, n, and p are each independently 0, 1, or 2.

42. The compound of claim 41, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3 Each is independently C1-C6 alkyl, halo, oxo or C1-C6 haloalkyl, and m, n and p are each independently 0 or 1.

43. The compound of claim 1, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA:

44. The compound of claim 43, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-1: Each R 4 is independently selected from the group consisting of H, C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, NH2 and oxo, and wherein q is 0, 1, 2 or 3.

45. The compound of claim 44, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-2: wherein K is a C1-C4 alkylene group optionally substituted by a halo group, a hydroxy group or a C1-C6 alkoxy group.

46. ​​The compound of claim 45, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-3: wherein K is a C1-C3 alkylene group optionally substituted by a hydroxy group.

47. The compound of claim 46, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-4a or IA-4b: wherein each X1 is independently selected from the group consisting of CH2, CH, O, S, N and NH.

48. The compound of claim 47, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-5a or IA-5b: wherein each X1 is independently selected from the group consisting of CH2, CH, NH, N and O.

49. The compound of claim 48, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound of formula IA-6a or IA-6b:

50. The compound of claim 49, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, which is a compound selected from the group consisting of Formula IA-7a, Formula IA-7b, Formula IA-7c, Formula IA-7d, Formula IA-7e, and Formula IA-7f: wherein each X2 is independently CH or N, and each s is independently 1, 2 or 3.

51. A compound as described in any one of claims 45-50, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein K is 52. A compound as described in any of claims 43-50, or a single stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein Y is a bond, NH, NH-(C1-C6 alkylene)-, N-(C1-C6 alkyl) or N-(C1-C6 alkylene-C3-C8 cycloalkyl).

53. The compound of any one of claims 43-50, or a single stereoisomer or mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein L is a bond or a C1-C6 alkylene.

54. A compound, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of 55. A compound, or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

56. A pharmaceutical composition comprising the compound according to any one of claims 1 to 55 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

57. A method for preparing a compound of formula I: or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, the process comprising: The compound of formula A: and: Formula B' (B') or formula C (C) Compound Combined under reductive amination conditions to provide the compound of formula I, Among them, ring A, ring B, ring C, J, L, R 1 , R 2 , R 3 , R x , R y , R x’ , R y’ , m, n and p are as defined in any one of claims 1 to 53; Ring B1 is a nitrogen-containing 3-8 membered monocyclic heterocycloalkylene or a nitrogen-containing 6-12 membered bicyclic heterocycloalkylene, each of which is optionally and independently substituted with up to three substituents selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halo, CN, C1-C6 haloalkyl, OH, COO(C1-C6 alkyl), CONH2, CONH(C1-C6 alkyl), CON(C1-C6 alkyl)2 and C1-C6 hydroxyalkyl; Y is a bond or a C1-C6 alkylene optionally substituted by OH, NH2, CN, halo or C1-C6 alkoxy, wherein one methylene unit of the C1-C6 alkylene is optionally replaced by NH, N-(C1-C6 alkyl), N-(C1-C6 hydroxyalkyl), N-(C1-C6 haloalkyl) or N-(C1-C6 alkylene-C3-C8 cycloalkyl); Y1 is a bond or C1-C5 alkylene optionally substituted with OH, NH2, CN, halo or C1-C6 alkoxy; and R z It is H, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 haloalkyl or C1-C6 alkylene-C3-C8 cycloalkyl.

58. A method for preparing a compound of formula ID: or a pharmaceutically acceptable salt thereof, the process comprising: The compound of formula E Coupling with a compound of formula F Among them, ring C, K, R 1 , R 2 , R 3 , R x , R y , m, n and p are as defined in any one of claims 1-53; and P is a hydroxy protecting group.

59. The process of claim 58, further comprising the step of removing the hydroxy protecting group to provide a compound of formula IE 60. The process of claim 59, further comprising the step of oxidizing the hydroxyl group to provide the compound of formula ID.

61. A compound of formula ID: or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein C, K, R 1 , R 2 , R 3 , R x , R y , m, n and p are as defined in any one of claims 1-53.

62. A compound of formula II: or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein Ring A, Ring B, Ring C, L, R 1 , R 2 , R 3 , R x’ , R y’ , m, n and p are as defined in any one of claims 1-53.

63. The compound of claim 62 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds 64. A compound of formula III or a single stereoisomer or a mixture of stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein ring B, ring C, L, R 1 , R 2 , R 3 , R x’ , R y’ , m, n and p are as defined in any one of claims 1-53.

65. The compound of claim 64 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds

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