MX2013005200A - Hydroxamic acid derivatives and their use in the treatment of bacterial infections. - Google Patents

Abstract

Antibacterial compounds of Formula I are provided: as well as stereoisomers and pharmaceutically acceptable salts thereof; pharmaceutical compositions comprising such compounds; methods of treating bacterial infections by the administration of such compounds; use of such compounds in the treatment of bacterial infections and processes for the preparation of such compounds.

Description

DERIVATIVES OF HYDROXAMIC ACID AND ITS USE IN THE TREATMENT OF BACTERIAL INFECTIONS Field of the Invention This invention relates in general to the treatment of infections caused by gram-negative bacteria. More specifically, the invention described herein relates to the treatment of gram-negative infections by inhibiting the activity of UDP-3-0- (R-3-hydroxydecanoyl) -N-acetylglucosamine deacetylase (LpxC). The present invention provides small molecule inhibitors of LpxC, pharmaceutical formulations containing these inhibitors, methods for treating patients with these pharmaceutical formulations, and methods for preparing these pharmaceutical formulations and inhibitors. The invention described herein relates to the treatment of gram-negative infections by administering compounds capable of inhibiting the activity of UDP-3-0- (R-3-hydroxydecanoyl) -N-acetylglucosamine-deacetylase (LpxC), either alone or in combination with a second antibacterial agent.

Background of the Invention During the past several decades, the frequency of antimicrobial resistance and its association with serious infectious diseases has increased at alarming rates. The problem of resistance Ref. 240984 antibacterial is combined by the existence of bacterial strains resistant to multiple antibacterials. In this way, there is a need for new antibacterials, particularly antibacterials with new mechanisms of action. A previously untapped but highly conserved target, LpxC, provides a new opportunity to develop small, antibacterial, broad-spectrum molecules that comprise a new class of active bactericidal chemical entities that must find little, if any, resistance related to target or target, which occurs naturally. LpxC (the enzyme uridyildiphospho-3-0- (R-hydroxydecanoyl) -N-acetylglucosamine-deacetylase) is present through all Gram-negative bacterial species of interest and is included in the first step involved in the biosynthesis of outer membrane. In this way, LpxC is essential for survival and presents an ideal target or target for antibiotic activity in Gram-negative bacteria species.

Investigators have identified some compounds with antibacterial activity that target target or target lipid biosynthesis. For example, Jackman et al. (J. Biol. Chem., 2000, 275 (15), 11002-11009); Yckoff et al. (Trends in Microbiology, 1998, 6 (4), 154-159); U.S. Patent Application Publication No. 2001/0053555 (published December 20, 2001, corresponding to PCT International Publication No. O 98/18754, published May 7, 1998); International PCT Publication No. WO 00/61134 (published October 19, 2000); U.S. Patent Application Publication No. 2004/0229955 (published November 18, 2004); and International PCT Publication No. WO 2008/154642 (published December 18, 2008) describe all compounds having anti-LpxC antibacterial activity. The commercial development of these LpxC inhibitors has been complicated by toxicity of these compounds in mammalian animals at concentrations at or near those required for antibacterial activity. The compounds presented herein are significantly better tolerated than other closely related compounds having anti-LpxC activity.

Although there have been advances in the field, the need remains for LpxC inhibitors that have activity as bactericidal agents against gram-negative bacteria and have an acceptable toxicity / tolerance profile. Accordingly, an object of this invention is to provide compounds and combinations of these compounds for use in the preparation of non-toxic antibacterials and other pharmaceuticals capable of inhibiting gram-negative bacterial infections. A further object of the present invention is to provide synergistic combinations of antibacterial agents with LpxC inhibitors., which have intrinsic antibacterial properties as well as the ability to improve the permeability of the outer membrane of gram-negative bacteria to other antibacterial agents. The use of synergistic combinations of drugs may have many advantages over conventional single-drug chemotherapy, including reduced side effects of drugs due to lower used doses or shorter treatment time, faster cure of infection by shortening hospital stays , increasing the spectrum of controlled pathogens, and decreasing the incidence of development of antibiotic resistance.

Brief Description of the Invention The present invention provides novel compounds, pharmaceutical formulations including the compounds, methods for inhibiting UDP-3-0- (R-3-hydroxydecanoyl) -N-acetylglucosamine deacetylase (LpxC), and methods for treating gram-negative bacterial infections.

In one aspect, the invention provides compounds of Formula I: and stereoisomers and pharmaceutically acceptable salts thereof, wherein A is a C3-C6-substituted cycloalkyl, wherein at least one substituent is a primary Ci-C3 alcohol; B is absent, -CH = CH-, -C = C- or is an unsubstituted phenyl; C is -CH = CH-, -C = C- or an unsubstituted phenyl, where if B is -CH = CH-, then C is also not -CH = CH-; R1, R2 and R3 are independently selected from hydrogen and Ci-C3-substituted and unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached form an unsubstituted C3-C3-cycloalkyl group, or R2 and R3 , together with the carbon atom and Q to which they are attached form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 carbon atoms, where 1-2 ring atoms of the heterocyclic ring of N, O and S are selected; and Q is O or NR, wherein R is hydrogen or a C1-C3-unsubstituted alkyl.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

In another aspect, the present invention provides a pharmaceutical composition comprising an effective amount of an antibacterial compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

In another aspect, the present invention provides a method for inhibiting a deacetylase enzyme in gram-negative bacteria, thereby affecting bacterial growth, which comprises administering to a patient in need of this inhibition an inhibitory compound of LpxC of Formula I or a stereoisomer or pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method for inhibiting LpxC in gram-negative bacteria, thereby modulating the virulence of a bacterial infection, which comprises administering to a patient in need of this inhibition an inhibitory compound of LpxC of Formula I or a stereoisomer or pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method for treating a subject having a bacterial infection comprising administering to the subject in need thereof an antibacterially effective amount of a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof. In a more specific mode of the treatment method, the bacterial infection is a Gram-negative bacterial infection. In these embodiments, the bacterium is Pseudo onas aeruginosa, Burkholderia (e.g., Burkholderia cepacia), Enterobacteriaceae, Franciscellaceae (e.g., Franciscella tularensis), Serratia, Proteus, Klebsiella, Enterobacter, Ci trobacter, Salmonella, Providence, Yersinia (e.g. Yersinia pestis), Morganella or Escherichia coli. In a particular embodiment the bacterium is Pseudomonas aeruginosa, Burkholderia, Franciscellaceae, Enterobacter, Yersinia or Escherichia coli. In this modality the bacterium is Pseudomonas aeruginosa. In another modality the bacterium is Escherichia coli. In another modality the bacterium is Stenotrophomonas maltophila, Alcaligenes xylosoxidans, Haemophilus, Neisseria species, Cedecea or Edwardsiella species. In a further specific embodiment, the subject is a mammal and in certain modalities he is a human.

One aspect of the invention provides pharmaceutical compositions comprising an LpxC inhibitor and a second antibacterial agent. In one implementation, the second antibacterial agent is selected from the group consisting of vancomycin, linezolid, azithromycin, imipenem, teicoplanin, daptomycin, clindamycin, rifampin, cefotaxime, gentamicin, novobiocin, and telavancin. In this implementation, the second antibacterial agent is vancomycin or rifampin. In another embodiment, the LpxC inhibitor is a compound of Formula I: or a stereoisomer or pharmaceutically acceptable salt thereof, wherein A is a substituted C3-C6-cycloalkyl, wherein at least one substituent is a primary Ci-C3 alcohol; B is absent, -CH = CH-, -C = C- or an unsubstituted phenyl; C is -CH = CH-, -C = C- or an unsubstituted phenyl, where if B is -CH = CH-, then C is also not -CH = CH-; R1, R2 and R3 are independently selected from hydrogen and Ci-C3-substituted or unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached, form an unsubstituted C3-Ce-cycloalkyl group, or R2 and R3 , together with the carbon atom and Q to which they are attached, form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 carbon atoms, wherein 1-2 ring atoms are selected from the heterocyclic ring of N, O and S; and Q is O or NR, wherein R is hydrogen or an unsubstituted C3-alkyl.

Another aspect of the invention provides methods for treating a patient with a gram-negative bacterial infection, comprising co-administering a synergistic amount, for example in a synergistic amount in vivo, of an antibacterial agent and an LpxC inhibitor of Formula I. In one implementation, the antibacterial agent is selected from the group consisting of yancomycin, linezolid, azithromycin, imipenem, teicoplanin, daptomycin, clindamycin, rifampin, cefotaxime, gentamicin, novobiocin, and telavancin. In this implementation, the antibacterial agent is vancomycin or rifampin.

These and other aspects of the invention will be apparent with reference to the following detailed description.

Brief Description of the Figures Figure 1 illustrates the in vivo synergy of compound 1-1 and vancomycin in bacterial strain ATCC 43816.

Detailed description of the invention The present invention provides novel compounds and methods for inhibiting LpxC in gram-negative bacteria, and novel methods for treating bacterial infections. The compounds provided herein may be formulated into pharmaceutical formulations and medicaments that are useful in the methods of the invention. The. invention also provides the use of the compounds in preparing pharmaceuticals and pharmaceutical formulations, for the use of the compounds in inhibiting LpxC, and the use of the compounds in the treatment of bacterial infections in a subject. The invention further provides compositions and methods for treating gram-negative infections by administering compounds capable of inhibiting the activity of UDP-3-0- (R-3-hydroxydecanoyl) -N-acetylglucosamine-deacetyl-asa (LpxC), either alone or in combination with the administration of a second antibacterial compound.

A. Definitions The following abbreviations and definitions are used throughout this application: "LpxC" is an abbreviation that means UDP-3-0- (R-3-hydroxydocanoyl) -N-acetylglucosamine-deacetylase.

As used herein, the following definitions should apply unless otherwise indicated.

. "Alkyl" refers to hydrocarbyl groups, aliphatic, saturated, monovalent having from 1 to 10 carbon atoms and preferably from 1 to 6 carbon atoms. This term includes, by way of example, linear and branched hydrocarbyl groups, such as methyl (CH3-), ethyl (CH3CH2-), n-propyl (CH3CH2CH2-), isopropyl ((CH3) 2CH-), n-butyl (CH3 CH2CH2CH2-), isobutyl ((CH3) 2-CHCH2-), sec-butyl ((CH3) (CH3CH2) CH-), t-butyl ((CH3) 3C-), n-pentyl (CH3CH2CH2CH2CH2-) and neopentyl ((CH3) 3CCH2-).

"Alkoxy" refers to the group -O-alkyl, where alkyl is as defined herein. Alkoxy includes methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like.

"Amino" refers to the group -NH2.

"Primary alcohol" refers to the group -alkyl-OH, wherein the hydroxyl radical is connected to a primary carbon. Examples include -CH2OH (hydroxymethyl), -CH2CH2OH (hydroxymethyl) and CH (CH3) CH2OH (l-hydroxypropan-2-yl).

"Alkenyl" refers to branched or straight chain hydrocarbyl groups having from 2 to 6 carbon atoms and preferably from 2 to 4 carbon atoms and having at least 1 and preferably from 1 to 2 sites of vinyl unsaturation (> C = C <). These groups are exemplified by vinyl, allyl, and but-3-en-1-yl. Included within this term are the cis and trans isomers or mixtures of these isomers.

"Alkynyl" refers to hydrocarbyl groups, monovalent, straight or branched having from 2 to 6 carbon atoms and preferably from 2 to 3 carbon atoms and having at least 1 and preferably from 1 to 2 sites of acetylenic saturation -C = C-. Examples of alkynyl groups include acetylenyl (-C = CH), and propargyl (CH2C = CH).

"Carboxyl" or "carboxy" refers to -COOH or salts thereof.

"Cyano" or "nitrile" refers to the -NC group.

"Cycloalkyl" refers to cyclic alkyl groups of 3 to 13 carbon atoms that they individually have. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like.

"Guanidino" refers to the group -NHC (= NH) H2.

"Halo" or "halogen" refers to fluoro, chloro, bromo, and iodo, and is typically fluoro or chloro.

"Hydroxy" or "hydroxyl" refers to the -OH group.

"Heterocycle", "heterocyclic" and "heterocyclyl" refer to a saturated or unsaturated group having an individual ring, and having 3 to 15 carbon atoms, including 1 to 4 heteroatoms. These ring atoms are selected from the group consisting of nitrogen, sulfur, or oxygen. In one implementation, the nitrogen and / or sulfur atoms of the heterocyclic group are optionally oxidized to provide the N-oxide, -S (0) -, or -S02- portions.

"Nitro" refers to the group -N02.

"Nitrous" refers to the group -NO.

"Oxo" refers to the atom (= 0).

"Substituted" refers to a group having one or more hydrogens replaced with substituents selected from the group consisting of alkoxy, acyl, acylamino, acyloxy, amino, aminocarbonyl, aminothiocarbonyl, aminocarbonyl-amino, aminothiocarbonylamino, aminocarbonyloxy, amidino, carboxyl, carboxyl ester, (carboxyl ester) amino, (carboxyl ester) oxy, cyano, guanidino, halo, hydroxy, nitro, S03H, sulfonyl, sulfonyloxy, thioacyl, thiol, and alkylthio, wherein these substituents are as defined in the present. In certain substituted cyclic groups, "substituted" also refers to a group having two hydrogens replaced with an individual double bond oxygen atom (an oxo group) or an individual double bond sulfur atom (thioxo). In some implementations, the substituted group has from 1 to 3 of the substituents mentioned above. In other implementations, the substituted group has from 1 to 2 of the substituents mentioned above.

"Sulfonyl" refers to the group -S02-alkyl, -S02-substituted alkyl, -S02-alkenyl, -S02-substituted alkenyl, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, and substituted alkynyl are as defined at the moment. Sulfonyl includes groups such as methyl-S02-.

"Sulfonyloxy" refers to the group -OS02-alkyl, -OS02-substituted alkyl, -OS02-alkenyl, -OS02-substituted alkenyl, -OS02-alkynyl, -OS02-substituted alkynyl, wherein alkyl, substituted alkyl, alkenyl, alkenyl substituted, alkynyl, and substituted alkynyl are as defined herein.

"Thioacyl" refers to the groups HC (S) -, alkyl-C (S) -, substituted alkyl-C (S) -, alkenyl-C (S) -, substituted alkenyl-C (S) -, alkynyl- C (S) -, and substituted alkynyl-C (S) -, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, and substituted alkynyl are as defined herein.

"Tiol" refers to the group -SH.

"Tioxo" refers to the atom (= S).

"Alkylthio" refers to the group -S-alkyl, wherein alkyl is as defined herein. In other implementations, sulfur may be oxidized to -S (0) -. The sulfoxide can exist as one or more stereoisomers.

Unless otherwise indicated, the nomenclature of the substituents that is not explicitly defined herein is achieved by naming the terminal portion of the functionality followed by functionality adjacent to the point of attachment. For example, the substituent "arylalkyloxycarbonyl" refers to the group (aryl) - (alkyl) -0-C (0) -.

In general, reference to a certain element such as hydrogen or H is proposed to include all isotopes of that element. For example, if a substituent group is defined to include hydrogen or H, it also includes deuterium and tritium.

The present invention also includes isotopically-labeled compounds of the present invention, which are structurally identical to those described herein, but by the fact that one or more atoms are replaced by an atom having an atomic mass or a mass number other than the Atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2H, 3H, .13C, 14C, 15N, 180, 170, 31P, 32P, 35S, 18F and 36C1, respectively. The compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of these compounds and the prodrugs that contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labeled compounds of the present invention, for example those into which radioactive isotopes such as 3H and 1C are incorporated, are useful in tissue distribution assays of drugs and / or substrate. Particular preference is given to tritiated isotopes, ie, 3H, and carbon-14, ie, 1C, for their ease of preparation and detectability. Additionally, replacement with heavier isotopes such as deuterium, i.e., 2H, can give certain therapeutic advantages that result in greater metabolic stability, for example increased in vivo half-life or reduced dose requirements and therefore, may be preferred in some circumstances The isotopically-labeled compounds of this invention and prodrugs thereof can be prepared in general by carrying out known or referenced processes and by replacing an easily available isotopically labeled reagent with a non-isotopically labeled reagent.

"Stereoisomer" and "stereoisomers" refer to compounds that have the same atomic connectivity but different atomic arrangement in space. Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers, and diastereomers.

"Tautomer" is. refers to alternating forms of a molecule that differs in the position of a proton, such as eno-keto and imine-enamine tautomers, or tautomeric forms of heteroaryl groups that contain a ring atom arrangement of -N = C (H ) -NH-, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. A person skilled in the art will recognize that other arrangements of a tautomeric ring atom are possible.

"Patient" refers to humans and non-human animals, especially mammals.

"Pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts of a compound, salts which are derived from a variety of organic and inorganic counterions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium , ammonium, tetraalkylammonium, and the like; and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, phosphate, sulfate and the like.

"Pharmaceutically effective amount" and "therapeutically effective amount" refer to an amount of a compound sufficient to treat a specified disorder or disease or one. or more of its symptoms and / or to prevent the occurrence of the disease or disorder.

The term "synergistic" or "synergistic" as used herein means that the combined effect of the compounds when used in combination is greater than the additive effects of the compounds when used individually. "Synergism" can be defined quantitatively as an index of fractional inhibitory concentration (FICI) of = 0.5, where FICI is defined as the sum of the fractional inhibitory concentrations (FIC, for its acronym in English) of the individual components in a combination of two compounds, and the FIC is defined as the ratio of the minimum inhibitory concentration (MIC) of the compound in the combination divided by the MIC of the compound alone: . M I Cfárma8 A in combo (M ICdr drug B in combo MICdrug A single / 1 + ÍCfá nmaco B only |) Alternatively, "synergism", more particularly "synergism in vivo", can be defined quantitatively as a decrease of at least twice in the static dose of the agents used in combination, compared to the LpxC inhibitor or second antibacterial agent alone. In certain cases, an agent alone can never reach a static dose. In these cases, a combination is synergistic if bacterial growth can be stopped (CFU load at 24 hours is identical to that measured at 0 hours after infection) by combined administration with two compounds that alone can not achieve equilibrium.

"Co-administration" can be in the form of an individual formulation (combination, for example, a compound of the present invention and a second antibacterial agent with pharmaceutically acceptable excipients, which optionally secrete the active ingredients into different mixtures of excipients designed to control independently their respective release rates and durations) or by independent administration of separate formulations containing the active agents. "Co-administration" also includes concurrent administration (administration of a compound of the present invention and a second antibacterial agent at the same time) and administration varied in time (administration of a compound of the present invention at a different time from that of second antibacterial agent), while the compound of the present invention as the second antibacterial agent are present in the body at therapeutically effective concentrations during at least partially overlapping times.

The term "antibacterial agent" refers to agents that have either bactericidal or bacteriostatic activity. The term "inhibit growth" indicates that the rate of increase in numbers of a population of a particular bacterium is reduced. In this way, the term includes situations in which the bacterial population increases, but at a reduced speed, as well as situations where population growth stops, as well as situations where the numbers of bacteria in the population are reduced or the population is still eliminated. If an enzyme activity assay is used to examine the inhibitors, modifications in uptake / effluvium, solubility, half-life, etc. can be made to the compounds in order to correlate enzyme inhibition with growth inhibition. The activity of antibacterial agents is not necessarily limited to bacteria, but may also encompass activity against parasites, viruses, and fungi.

Unless the context requires otherwise, throughout the specification and claims that follow, the word "comprises" and variations thereof, such as, "comprises" and "comprising" will be understood in a sense open, inclusive, that is, as "including, but not limited to".

Reference throughout this specification to "one modality" or "modality" means that a particular feature, structure or feature described in conjunction with the embodiment is included in at least one embodiment of the present invention. In this way, the occurrences of the phrases "in one modality" or "in one modality" in several places throughout this specification do not necessarily refer to the same modality. Additionally, the features, structures, or particular features may be combined in any suitable manner in one or more embodiments.

B. Compounds, Compositions and Uses of the same In one aspect, the present invention provides compounds of Formula I: and stereoisomers and pharmaceutically acceptable salts thereof, wherein A is a C3-C6-substituted cycloalkyl, wherein at least one substituent is a primary C1-C3- alcohol; B is absent, -CH = CH-, -C = C- or an unsubstituted phenyl; C is -CH = CH-, -C = C- or an unsubstituted phenyl, where if B is - CH = CH -, then C is also not - CH = CH -; R1, R2 and R3 are independently selected from hydrogen and Ci-C3-substituted or unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached, form an unsubstituted C3-C6-cycloalkyl group, or R2 and R3 , together with the carbon atom and Q to which they are attached form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 carbon atoms, wherein 1-2 ring atoms of the heterocyclic ring of N, O and S are selected; and Q is 0 or NR, wherein R is hydrogen or a C1-C3-unsubstituted alkyl.

In certain modalities, Q is NR, and in some modalities Q is NH or NCH3. In certain embodiments, R1, R2, and R3 are independently selected from hydrogen and Ci-C3-substituted or unsubstituted alkyl, and in some embodiments are selected from hydrogen and Ci-C3-unsubstituted alkyl. In certain embodiments, R1 and R2 independently are C: i.-C3-unsubstituted alkyl. In certain embodiments, A is a C3-C6-cycloalkyl substituted with hydroxymethyl. In other modalities, both B and C are -C = C-.

One aspect of the invention provides compounds of the formula IA: and stereoisomers and pharmaceutically acceptable salts thereof, wherein A is a C3-C6-substituted cycloalkyl, wherein at least one substituent is a primary C1-C3- alcohol; R1, R2 and R3 are independently selected from hydrogen and C; i.-C3-substituted or unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached form a C3-C6-cycloalkyl group, or R2 and R3, together with the carbon atom and nitrogen to which they are attached, form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 ring atoms, wherein 1-2 ring atoms of the heterocyclic ring are selected from N, O and S; and R is hydrogen or an unsubstituted C3-alkyl. In certain embodiments, R1, R2 and R3 are independently selected from hydrogen and Ci-C3-substituted or unsubstituted alkyl, and in some embodiments are selected from hydrogen and C1-C3-unsubstituted alkyl. In one embodiment, A is a C3-C6-cycloalkyl, mono-substituted with a primary alcohol of Ci-C3-.

The compounds of the present invention include those listed in Table I. Certain compounds illustrated in Table 1 represent mixtures of two diastereomers. In this case, the notation "*" indicates that the two portions joined to the cyclic portion are trans to each other. The notation "#" indicates that the two portions attached to the cyclic portion are cis to each other. 10 5 10 fifteen ?? The compounds of the present invention can be easily synthesized using the methods described herein, or other methods that are well known in the art, For example, the synthesis of hydroxamic acids or similar molecular nuclei having a wide variety of substituents is reviewed comprehensively in Kline, T., et al., "Potent, novel in vitro inhibitors of the Pseudomonas aeruginosa deacetilase LpxC" J. Med Che. 2002, 45 (14), 3112-29; U.S. Patent No. 5,925,659; Pirrung, M. C, et al., "A Convenient Procedure for the Preparation of Amino Acid Hydroxamate from Esters" J. Org. Chem. 1995, 60, 8084-8085; Nhu, K., et al., "A New and Efficient Solid Phase Synthesis of Hydroxamic Acids" J. Org. Chem. 1997, 62, 7088-7089; PCT International Publication No. O 98/18754; Mellor, S.L., et al., "N-Fmoc-aminoxy-2-chlortrityl Polistyrene Resin: A Facile Solid-phase Methodology for the Synthesis of Hydroxamic Acids" retrahedron Lett. 1997, 38, 3311-3314; Khan, S. I., et al. , "A Facile and Convenient Solid-phase Procedure for Synthesizing Nucleoside Hydroxamic Acids" Tetrahedron. Lett. 1998, 39, 8031-8034; Zhang, Y., et al., "Design, Combinatorial Chemical Synthesis, and in vitro Characterization of Novel Urea Based Gelatinase Inhibitors" Bioorg. Med. Chem. Lett. 1999, 9, 2823-2826; Ito, Y., et al., "Synthetic Reactions by Complex Catalysts, XXXI, A Novel and Versatile Method of Heterocycle Synthesis" J. Am Chem. Soc. 1973, 95, 4447-4448; Ito, Y., et al., "Synthetic Reactions by Complex Catalysts XXXV" Syn. Commun. 1974, 4, 97-103; Witte, H. , et al., "Cyclische Imidsaurester aus Nitrilen und Aminoalkoholen" Liebigs Ann. Chem. 1974, 996-1009; Pattenden, G., et al., "Naturally Occurring Linear Fused Thiazoline-Thiazole Containing Metabolites: Total Synthesis of (-) Didehydromirabazole A, to Cytotoxic Alkaloid from Blue-Green Algae" J. Chem. Soc. Perkin Trans 1993, 1, 1629-1636; Boyce, R. J., et al., "Total Synthesis of Thiangazole, A Novel Naturally Occurring HIV-1 Inhibitor frora Poliangium sp." Tetrahedron 1995, 51, 7321-7330; Galeotti, N., et al., "Synthesis of Peptidyl Aldehydes from Thiazolines" Tetrahedron. Lett. 1997, 38, 2459-2462; Charette, AB, et al., "Mild Method for the Synthesis of Thiazolines from Secondary and Tertiary Amides" J. "Org. Chem. 1998, 63, 908-909; Bergeron, RJ, et al.," Effects of C- 4 Stereochemistry and C-41 Hydroxylation on the Iron Clearing Efficiency and Toxicity of Desferrithiocin Analogues "J. Med. Chem. 1999, 42, 2432-2440; Raman, P., et al.," Titanium (IV) -mediated Tandem Deprotection -cyclodehydration of Protected Cysteine N-Amides: Biomimetic Synthesis of Thiazoline- and Thiazole-containing Heterocycles "Org. Lett., 2000, 2, 3289-3292; Fernandez, X., et al.," Novel Synthesis of 2-Thioazolines "Tetrahedron Lett 2000, 41, 3381-3384; and Wlpf, P., et al., "C. Thiolysis of Oxazolinenes: A New, Selective Method for the Direct Conversion of Peptide Oxazolines into Thiazolines "Tetrahedron Lett, 1995, 36, 6395-6398, which are incorporated herein by reference.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

In another aspect, the invention provides a method for inhibiting a deacetylase enzyme in gram-negative bacteria, thereby affecting bacterial growth, which comprises administering to a patient in need of this inhibition a compound of Formula I or a stereoisomer or pharmaceutically salt acceptable of it.

In another aspect, the invention provides a method for inhibiting LpxC in gram-negative bacteria, thereby modulating the virulence of a bacterial infection, which comprises administering to a patient in need of this inhibition a compound of Formula I or a stereoisomer or pharmaceutically salt acceptable of it. In certain embodiments of the method for inhibiting LpxC using a compound of the present invention, the IC50 value of the compound is less than or equal to 10 uM with respect to LpxC. In other embodiments, the IC50 value is less than or equal to 1 μ ?, is less than or equal to 0.1 μ ?, is less than or equal to 0.050 μ ?, is less than or equal to 0.030 μ ?, is less that or equal to 0.025 μ ?, or is less than or equal to 0.010 μ ?.

In another aspect, the invention provides a method for treating a subject having a gram-negative bacterial infection comprising administering to the subject in need thereof an antibacterially effective amount of a compound of Formula I or a stereoisomer or pharmaceutically acceptable salt thereof. In this embodiment, the bacterium is Pseudomonas aeruginosa, Burkholderia (e.g., Burkholderia cepacia), Enterobacteriaceae, Franciscellaceae (e.g., Franciscella tularensis), Serratia, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providence, Yersinia (e.g., Yersinia pestis), Morganella or Escherichia coli. In a particular embodiment, the bacterium is Pseudomonas aeruginosa, Burkholderia, Franciscellaceae, Enterobacter, Yersinia or Escherichia coli. In this modality, the bacterium is Pseudomonas aeruginosa. In another embodiment, the bacterium is Escherichia coli. In another embodiment, the bacterium is Stenotrophomonas aitophila, Alcaligenes xyiosoxidans, Haemophilus, Neisseria species, Cedecea or Edwardsiella species.

In certain modalities, the subject can be a mammal, and in some modalities, he is a human.

Bacterial infections susceptible to treatment according to the present invention include primary infections and co-infections caused by a species of bacteria and one or more additional infectious agents such as, for example, bacteria, viruses, parasites and fungi.

The compounds of the invention can be used to treat conditions caused by the bacterial production of endotoxin, and in particular, by gram-negative bacteria and bacteria that use LpxC in the biosynthesis of lipopolysaccharide (LPS) or endotoxin.

The compounds of the invention are also useful in the treatment of conditions that are caused or exacerbated by the bacterial production of lipid A and LPS or endotoxin, such as sepsis, septic shock, systemic inflammation, localized inflammation, chronic obstructive pulmonary disease (COPD, by its acronym in English) and acute exacerbations of chronic bronchitis (AECB, for its acronym in English). For these conditions, the treatment includes the administration of a compound of the invention, or a combination of compounds of the invention, optionally with a second agent wherein the second agent is a second antibacterial agent or a non-antibacterial agent.

For sepsis, septic shock, systemic inflammation, localized inflammation, chronic obstructive pulmonary disease (COPD) and acute exacerbations of chronic bronchitis (AECB), representative non-antibacterial agents include antiendotoxins that include antibodies that bind to the endotoxin receptor, antibodies that are they bind to endotoxin, antiprotein antibodies that bind to CD14, antiprotein antibodies that bind to lipopolysaccharide and tyrosine kinase inhibitors.

In the treatment of serious or chronic infections of the respiratory tract, the compounds of the present invention can also be used with non-antibacterial agents administered by inhalation. Representative non-antibacterial agents used in this treatment include anti-inflammatory steroids, non-steroidal anti-inflammatory agents, bronchodilators, mucolytics, anti-asthma therapeutics and pulmonary fluid surfactants. In particular, the non-antibacterial agent can be albuterol, salbuterol, budesonide, beclomethasone, dexamethasone, nedocromil, beclomethasone, fluticasone, flunisolide, triamcinolone, ibuprofin, rofecoxib, naproxen, celecoxib, nedocromil, ipratropium, metaproterenol, pirbuterol, salmeterol, formoterol, indacaterol , bronchodilators, mucolytics, calfactant, beractant, poractant alfa, surfaxin or pulmozyme (also called domase-alpha).

The compounds of the invention can be used alone or in combination with a second antibacterial agent for the treatment of a serious or chronic infection of the respiratory tract including serious nosocomial and pulmonary infections such as those caused by Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Klebsiella. pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Serratia marcescens, Stenotrophomonas altophilia, Pseudomonas aeruginosa, Burkholderia cepacia, Alcaligenes xylosoxidans, Flavobacterium meningosepticum, Providencia stuartii and Citrobacter freundi, community lung infections such as those caused by Haemophilus Influenzae, Legionella species, Moraxella catarrhalis, Branhamella catarrhalis, Enterobacter species, Klebsiella species, and Proteus species, infections caused by other bacterial species such as Neisseria species, Shigella species, Salmonella species, Helicobacter pylori species, Vibrionaceae and Bordet it, as well as infections caused by Brucella species, Francisella tularensis and / or Yersinia Pestis.

When used to treat subjects infected with gram-negative bacterial infections, the compounds of the present invention can be used to sensitize gram-negative bacteria to the effects of a second bacterial agent.

The present invention provides novel combinations of compounds that include a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, as well as methods for treating subjects infected with gram-negative. The new combinations provided herein may be formulated into pharmaceutical formulations and medicaments that are useful in the methods of the invention. The invention also provides the use of the novel combinations in the preparation of medicaments and pharmaceutical formulations, for the use of the combinations in the treatment of bacterial infections in a patient.

A classical method for assessing synergies, referred to as the checkerboard type test, is used to predict the efficiency of antibacterial agents, and is described by Scribner et. al., (1982, Antimicrobial Agents and Chemotherapy (6): 939-943) and Goodman & Gilman (1980, The Pharmacological Basis of Therapeutics, Sixth Edition, pp. 1097-1098). The chessboard-type test comprises twice-in-series dilutions of the antibiotics individually and in combination in broth, which is then inoculated with the organism to be tested. After incubation, the minimum inhibitory concentration (MIC) of each drug used individually and in combination is determined (NB, the MIC is the lowest concentration of the drug that inhibits growth in the medium) . Synergism is indicated by a decrease in the MIC of each drug when used in combination. Antagonism is indicated by an increase in MIC of either or both drugs when used in combination.

Alternate methods for assessing synergy are reviewed in Greco, et al., Pharmacological Reviews 47 (2): 331-285 (1995), incorporated herein by reference in its entirety.

However, a positive result in a chessboard type test, ie, indicating synergism below the MIC, does not necessarily result in synergistic behavior in vivo. U.S. Patent Application Publication No. 2004-229955A1 reports strong synergy between erythromycin and an LpxC inhibitor, N- [(1S) -1- (aminomethyl) -2- (hydroxyamino) -2-oxoethyl] - 4- (4- { 4- [( { [(3-methylphenyl) methyl] amino.}. Acetyl) amino] phenyl.} Buta-1,3-diinyl) benzamide against the E. coli strain ATCC 25922. International PCT Patent Application No. PCT / US2010 / 33910 demonstrated that the combination of erythromycin and a variety of LpxC inhibitors does not show synergy in vivo.

LpxC, an essential gene in gram-negative bacteria, codes for the enzyme uridyldiphospho-3 -? - (R-hydroxydecanoyl) -N-acetylglucosamine-deacetylase. This enzyme catalyzes an early committed step in the biosynthesis of lipid A, the lipid portion of lipopolysaccharide that is an essential component of all gram-negative bacteria. Above the MIC, the LpxC inhibitor is expected to break the outer membrane, thereby allowing other antibacterial compounds to penetrate the outer membrane. Once these agents have penetrated the outer membrane, they can affect the periplasmic targets as is the case with vancomycin, or they can then diffuse through the inner membrane to interact with an intracellular target or target such as the ribosome (Erythromycin) or RNA polymerase (Rifampin). In the absence of an LpxC inhibitor, the ability of agents such as vancomycin to access its target or target is decreased by the outer membrane for the most part. Thus, without being bound by theory, it is believed that the biochemical mechanism that may be the basis of the observed synergy is the improved permeability of the outer membrane to agents such as vancomycin when combined with LpxC inhibitors.

In one modality, the. second antibacterial agent used in combination with a compound of Formula I, or stereoisomer or pharmaceutically acceptable salt thereof, is vancomycin, linezolid, azithromycin, imipenem, teicoplanin, daptomycin, clindamycin, rifampin, cefotaxime, gentamicin, novobiocin or telavancin. In this modality, the second antibacterial agent is vancomycin, teicoplanin, rifampin, azithromine, telavancin or novobiocin. In this modality, the second antibacterial agent is vancomycin or rifampin. In some embodiments of the invention, the second antibacterial agent and / or the compound of Formula I, or stereoisomer or pharmaceutically acceptable salt thereof, is administered a subtherapeutic dose, wherein a subtherapeutic dose is a dose that It will be insufficient to treat bacterial infections, if administered alone.

The pharmaceutical compositions of the present invention comprise a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, formulated in conjunction with one or more pharmaceutically acceptable carriers or diluents. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert, solid, semi-solid or liquid filler, diluent, encapsulating material or auxiliary formulation of any type. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose or sucrose starches such as corn starch and potato starch; cellulose and its talc derivatives such as sodium carboxy-cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; jelly; talcum powder; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; Sesame oil; olive oil; corn oil and soybean oil; glycols; such as polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar, buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic outlet solution; Ringed solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and flavoring agents, preservatives and Antioxidants may also be present in the composition, according to the formulator's judgment. The pharmaceutical compositions of this invention can be administered to humans and other animals orally, rectally, parenterally (as by intravenous, intramuscular or subcutaneous injection), intracisternally, intravaginally, intraperitoneally, topically (as by powders). , ointments or drops), buccally, or as an oral spray or a salt, or a liquid spray or dry powder formulation for inhalation.

Liquid dosage forms for oral administration include paceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate. , benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, peanut, corn, germ, olive, resinous and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to the inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and flavoring agents.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suitable suspending agents. The sterile injectable preparation can also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, 1% lidocaine, isotonic sodium chloride solution U.S.P. In addition, non-volatile insipid oils are used conventionally as a solvent or suspension medium. For this purpose any insipid non-volatile oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectable products.

Injectable formulations can be sterilized, for example, by filtration through a filter that retains bacteria, or by incorporating sterilizing agents in the form of solid or sterile compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of a drug, it is often desirable to slow the absorption of the subcutaneous or intramuscular injection drug. This can be achieved by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of drug absorption then depends on its rate of dissolution which, in turn, may depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be achieved by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microcapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending on the drug to polymer ratio and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations can also be prepared by trapping the drug in liposomes or microemulsions that are compatible with body tissues.

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

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one. excipient or pharmaceutically acceptable carrier, inert such as sodium citrate or dicalcium phosphate and / or) filling agents or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose , alginates, gelatin, polyvinylpyrrolidinone, sucrose and acacia gum, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato starch or tapioca, alginic acid, certain silicates, and sodium carbonate , e) solution retarding agents such as paraffin f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, acetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and ) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

Solid compositions of a similar type can also be employed as fillers in hard and soft gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and may also be of a composition that they release the active ingredients only, or preferentially in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include waxes and polymeric substances.

Solid compositions of a similar type can also be employed as fillers in hard and soft fill gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The antibacterial compounds may also be in microencapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In these solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. These dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, for example, tabletting lubricants and other tabletting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents. They may optionally contain opacifying agents and may also be of a composition that they release the active ingredients only, preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of this invention include ointments, paste, creams, lotions, gels, powders, solutions, sprays, inhalations or patches. The active component is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservative or buffer as may be required. Ophthalmic formulations, ear drops and the like are also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain in addition to an active compound of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, Talc and zinc oxide or mixtures of these.

The compositions of the invention can also be formulated for distribution as a liquid aerosol or inhalable dry powder. Liquid aerosol formulations can be nebulized predominantly in particle sizes that can be distributed to the terminal and respiratory bronchi where the bacteria reside in patients with bronchial infections, such as chronic bronchitis and pneumonia. Pathogenic bacteria are commonly present throughout the airways to the bronchi, brochioles, and parenchyma of the lung, particularly in terminal and respiratory bronchioles. During the exacerbation of the infection, the bacteria may also be present in the alveoli. The inhalable and liquid aerosol dry powder formulations are preferably distributed throughout the length of the endobronchial tree to the terminal bronchioles and eventually to the parenchymal tissue.

The aerosol formulations of the invention can be dispensed using an aerosol forming device, such as a jet, vibrating porous plate or ultrasonic nebulizer, preferably selected to allow the formation of aerosol particles having an average mass diameter predominantly between 1 and 5 pm. Additionally, the formulation preferably has ionic concentration of balanced osmolarity and balanced chloride concentration, and the smallest aerosolized volume capable of distributing the effective dose of the compounds of the invention to the site of infection. Additionally, preferably, the aerosol formulation does not negatively impair the functionality of the airways and does not cause undesirable side effects.

Aerosol devices suitable for administration of the aerosol formulation of the invention include, for example, jet, vibrating porous plate, ultrasonic nebulizers and dry powder energized inhalers, which are capable of nebulizing the formulation of the invention in one size of aerosol particle predominantly in the size range of 1-5 pm. Predominantly in this application means that at least 70% but preferably more than 90% of all generated aerosol particles are in a range of 1 to 5 m. A jet nebulizer works by air pressure to break a liquid solution into aerosol droplets. Vibrating porous plate nebulizers work by using a sonic vacuum produced by a porous plate that vibrates rapidly to extrude a drop of solvent through a porous plate. An ultrasonic nebulizer works by a piezoelectric crystal that cuts a liquid into small aerosol droplets. A variety of suitable devices are available, including, for example, AeroNew and AeroDose vibrating porous plate nebulizers (AeroGen, Inc., Sunnyvale, Calif.), Sidestream7 nebulizers (Medic-Aid Ltd., West Sussex, England), nebulizers of jet Pari LC7 and Pari LC Star7 Net (Pari Respiratory Equipment, Inc., Richmond, Va.), and aerospace untrasonic nebulizers (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and UltraAire7 (Omron Healthcare, Inc., Vernon Hills, III).

The compounds of the invention may also be formulated for use as topical powders and sprays which may contain in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of this substance. The sprays may additionally contain customary propellants such as chlorofluorohydrocarbons.

Transdermal patches have the additional advantage of providing controlled distribution of a compound to the body. These dosage forms can be made by dissolving or dispensing the compound in the appropriate medium. Absorption enhancers may also be used to increase the flow of the compound through the skin. The speed can be controlled either by providing a 4o speed controlling membrane by dispersing the compound in a polymer matrix or gel.

According to the methods of treatment of the present invention, bacterial infections are treated or prevented in a patient such as a human or a lower mammal by administering to the patient a therapeutically effective amount of a compound of Formula I, or a stereoisomer or salt pharmaceutically acceptable thereof, in amounts and times as necessary to achieve the desired result. By a "therapeutically effective amount" of a compound of the invention is meant a sufficient amount of the compound to treat bacterial infections, at a reasonable benefit / risk ratio applicable to any medical treatment. However, it will be understood that the total daily dose of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific, therapeutically effective dose level for any particular patient will depend on a variety of factors including the disorder that the severity of the disorder is treated.; the activity of the specific compound used, - the specific composition used; age, body weight, general health, gender and diet of the patient; the administration time, route of administration, and rate of excression of the specific compound employed; the duration of the treatment; drugs used in combination or coincident with the specific compound used; and similar factors well known in the medical arts.

The total daily dose of the compounds of this invention administered to a human or other animal in individual or divided doses may be in amounts, for example, from 0.01 to 200 mg / kg of body weight or more usually from 0.1 to 50 mg / kg of body weight. In certain embodiments, the total daily dose administered to a human or other mammal is from 1.0 to 100 mg / kg of body weight or from 5.0 to 25 mg / kg of body weight. The individual dose compositions may contain these amounts or submultiples thereof to constitute the daily dose. In general, treatment regimens according to the present invention comprise administering to a patient in need of this treatment from about 10 mg to about 15 g of the compounds of this invention per day in single or multiple doses, more usually, from 100 mg to 5 g, and even more usual from 250 mg to 1 g per day in single or multiple doses.

Methods of formulation are well known in the art and are described, for example, in Remington: The Science and Practice of Pharmacy, Mack Publishing Company, Easton, Pa. , 19th Edition (1995). The pharmaceutical compositions for use in the present invention may be in the form of sterile non-pyrogenic liquid solutions or suspensions, coated capsules, suppositories, lyophilized powders, transdermal patches and other forms known in the art.

A "kit" as used in the present application includes a container for containing the pharmaceutical compositions and may also include divided containers such as a divided bottle or a divided laminated container. The container may be in any conventional form or conventional form as is known in the art to be made of a pharmaceutically acceptable material, for example a paper or cardboard box, a glass or plastic bottle or jar, a resealable bag (for example example, to retain a "filling" of tablets for placement in a different container), or a blister pack with individual doses to press the package according to a therapeutic program. The container may depend on the exact dosage form comprised, for example, a conventional cardboard box will not generally be used to maintain a liquid suspension. It is feasible that more than one container can be used in conjunction with an individual container to market an individual dosage form. For example, the tablets may be contained in a bottle which in turn is contained within a box.

An example of this kit is a so-called blister pack. Blister packs are well known in the packaging industry and are widely used for the packaging of pharmaceutical unit dosage forms (tablets, capsules, and the like). The blister packs generally consist of a sheet or sheet of relatively rigid material covered with a sheet of a preferably transparent plastic material. During the packaging process, depressions are formed in the sheet or sheet of plastic. The depressions have the size and shape of the individual tablets or capsules to be packaged or they may be of the size and shape to accommodate multiple tablets and / or capsules to be packaged. Then, the tablets or capsules are placed in the depressions accordingly and the sheet or sheet of relatively rigid material is sealed against the sheet of plastic on the surface of the sheet that is opposite from the direction in which the depressions were formed. As a result, the tablets or capsules are individually sealed or collectively sealed, as desired, in the depressions between the plastic sheet and the sheet or sheet. Preferably, the strength of the sheet or sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure in the depressions whereby an opening is formed in the sheet at the depression site. The capsule or tablet can then be removed through the opening.

It may be desirable to provide a written memory aid, where the written memory aid is of the type that contains information and / or instructions for the practitioner, pharmacist or other health care provider, or subject, for example, in the form of numbers following the tablets or capsules so the numbers correspond to the days of the regimen that should be taken the tablets or capsules that are specified or a card that contains the same type of information. Another example of this memory aid is a calendar printed on a card, for example, as follows "first week, Monday, Tuesday," .... etc "second week, Monday, Tuesday,", etc. Other variations of memory aids will be readily apparent. An "individual dose" can be a single tablet or capsule or several tablets or capsules taken on a given day. When the kit contains separate compositions, a daily dose of one or more compositions of the kit may consist of a tablet or capsule while a daily dose of another or more compositions of the kit may consist of several tablets or capsules.

Another specific modality of a kit is a dispenser designed to disperse the daily doses one at a time in the order of their intended use. Preferably, the dispenser is equipped with a memory aid, to further facilitate compliance with the regime. An example of this memory aid is a mechanical counter that indicates the number of daily doses that have been dispensed. Another example of this memory aid is a battery-powered microchip memory coupled with a liquid crystal reader, or audible reminder signal which, for example, reads the date that the last daily dose was taken and / or remembers when will take the next dose.

The kits of the present invention may also include, in addition to a compound of the present invention, one or more additional pharmaceutically active compounds. For example, the additional compound is a second antibacterial. The additional compounds can be administered in the same dosage form as the compound of the present invention or in a different dosage form. Likewise, the additional compounds can be administered at the same time as the compound of the present invention or at different times.

The compositions of the present compounds can also be used in combination with other known antibacterial agents of similar spectrum to (1) improve the treatment of severe gram-negative infections covered by the spectrum of this compound or (2) to add coverage in severe infections in the which multiple organisms are suspected in which another agent of a different spectrum may be required in addition to this compound. Potential agents include members of the aminoglycosides, penicillins, cephalosporins, fluoroquinolones, macrolides, glycopeptides, lipopeptides, and oxazolidinones. The treatment may comprise administering a composition having both a compound of the present invention and a second antibacterial compound or administration of a compound of the present inventive compounds followed by or preceded by the administration of a second antibacterial agent.

The foregoing can be better understood by reference to the following examples, which are presented for illustration and not to limit the scope of the inventive concepts.

And emplos With reference to the following examples, the compounds of the present invention were characterized by high performance liquid chromatography (HPLC) using a Waters Millenium chromatography system with a Separation Module 2690 (Milford, Mass.) . The analytical columns were Alltima C-18 reverse phase, 4.6x250 mm Alltech (Deerfield, III.). A gradient elution was used, typically starting with 5% acetonitrile / 95% water and progressing to 100% acetonitrile over a period of 40 minutes. All solvents contained 0.1% trifluoroacetic acid (TFA). The compounds were detected by absorption of ultraviolet (UV) light at either 220 or 254 nm. In some cases, purity was assessed by thin layer chromatography (TLC) using silica gel plates with glass or plastic backing, such as, for example, flexible sheets Baker-Flex Silica Gel 1 B2-F. TLC results were easily detected visually under ultraviolet light or by using well known iodine vapor staining techniques and various other staining techniques.

Mass spectrometric analysis was performed on one of two LCMS instruments: a Waters System. (Alliance HT HPLC and a Micromass ZQ mass spectrometer; Column: Eclipse XDB-C-18, 2.1x50 mm; solvent system: 5-95% (or 35-95%, or 65-95% or 95-95% ) acetonitrile in water with 0.05% TFA flow rate of 0.8 mL / min, molecular weight range 500-1500, cone speed 20 V, column temperature at 40 ° C.) or a Hewlett Packard System (Series 1100 HPLC); Column: Eclipse XDB-C18, 2.1x50 mm, solvent system: 1-95% acetonitrile in water with 0.05% TFA, flow rate 0.4 mL / min, molecular weight range 150-850, cone voltage 50 V, column temperature 30 ° C). All masses are reported as those of protonated origin ions.

The GCMS analysis was performed on an instrument Hewlett Packard (HP6890 series gas chromatograph with a 5973 Selective Mass Detector, injector volume: 1 L, initial column temperature: 50 ° C, final column temperature: 250 ° C, increase tempo: 20 minutes flow rate of gas: 1 mL / min, - column: 5% phenyl-methyl-siloxane, Model #HP 190915-443, dimensions: 30.0 mx25 mx0.25 m).

Nuclear magnetic resonance (RM) analysis was performed with a 300 MHz NMR (Palo Alto, Calif.). The spectral reference was either TMS or the known chemical change of the solvent. Some samples of compounds were run at elevated temperatures (e.g., 75 ° C) to promote increased sample solubility. purity of some of the invention compounds was assessed by experimental analysis (Desert Analytics, Tn, Ariz.) determined melting points in an apparatus Laboratory Devices Mel-Temp (Holliston, Mass.).

The preparative separations were carried out using a Flash 40 and KP-Sil chromatography system, 60A (Biotage, Charlottesville, Va.), Or by flash column chromatography using silica gel packing material (230-400 mesh) or HPLC using a C-18 reversed phase column. The typical solvents used for the Flash 40 Biotage system and the flash column chromatography were dichloromethane, methanol, ethyl acetate, hexane, acetone, aqueous hydroxylamine and triethylamine. The typical solvents used for reverse phase HPLC were varying concentrations of acetonitrile and water with 0.1% trifluoroacetic acid.

A. Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl-4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) enzamide (1-1) 1 2 3 Methyl 2-ethynylcyclopropanecarboxylate (2) Racemic ethyl 2-formylcyclopropanecarboxylate 1 (10 g, 70.3 mmol) and Bestmann Ohira reagent (16.4 g, 85 mmol) were dissolved in anhydrous methanol (100 ml) under N2. Potassium carbonate (19.4 g, 141 mmol) was then slowly added in portions and the solution was stirred for 18 h. The solvents were removed under pressure, reduced to 20 ° C, water (100ml) was added and the product was extracted with dichloromethane (2x 200ml), dried over sodium sulfate, and slowly concentrated to give 2-ethynylcyclopropane carboxylate. racemic methyl (4.16g) which was confirmed by NMR. (2-Ethynylcyclopropyl) methanol (3) Then lithium borohydride (175mg, 8.06mmol) was added slowly to racemic methyl 2-ethylcyclopropanecarboxylate (lg, 20.6mmol) in anhydrous THF (20ml) under nitrogen and stirring was continued for 2h. The reaction mixture was quenched with a few drops of acetic acid and the solvent was removed. The crude product was extracted with ethyl acetate (2x50ml), dried over sodium sulfate and concentrated slowly to give racemic (2-trans-ethynylcyclopropyl) methanol, 3, as a yellow liquid (735mg) and used as such to the next step. 3- (tert-Butoxycarbonylamino) -2- (4- ((2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (2S) -methyl ester (5) CuCl (42mg, 0.416 mmol) was added slowly to a stirred solution of racemic (2-trans-ethynylcyclopropyl) methanol (400mg, 4.16mmol), 2- (S) - (4- (bromoetinyl) -benzamido) -3- ( methyl tert-butoxycarbonylamino) -3-methylbutanoate (1.9g, 4.16mmol) in a mixture of THF (20ml), methanol (10ml) and butyl-amine (10ml) under nitrogen followed by a few crystals of hodrixylamine hydrochloride (30mgs) ). Stirring was continued for 4 h. The solvent was removed under reduced pressure, water (100ml) was added and it was extracted with ethyl acetate (2xl50ml), dried over sodium sulfate, concentrated and purified in ISCO to give 4 2mg of 3- (tert-butoxycarbonylamino) -2- (4- (2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (2S) -methyl ester. 1-1: N- ((S) -3-amino-l- (hydroxy-amino) -3-methyl-l-oxobutan-2-yl) -4- (((1, 2trans) -2- (hodriximetll) cyclopropyl) buta-1, 3-diinyl) benzamide 3- (tert-Butoxycarbonylamino) -2- (4- ((2- (hydroxy-methyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanedioate (2-S) -methyl 5 ( 0.370g, 0.814mmol) was taken in dichloromethane (25ml), treated with TFA (2ml) and stirred for 20min. Excess solvent and TFA were removed under reduced pressure to give deprotected material that was re-dissolved in IPA (10mL) and treated with aqueous hodrixylamine (50%, 2mL, excess) and kept in the refrigerator for 2 days. The excess solvent was removed under reduced pressure and the crude product was purified by reverse phase HPLC to give 42mgs of 1-1 as its trifluoroacetate salt. LC-MS (M + l) 370, Chemical Formula: C20H23 3O4, Exact Mass: 369.17. RMIS ^ H (DMSO-d6), TFA salt: 0.9 (m, 2H) 1.24 (s, 3H), 1.29 (s, 3H), 1.40 (m, 1H), 3.24 (m, 1H), 3.4 (m , 1H), 4.64 (d, 1H), 4.69 (d, 1H), 7.61 (d, 2H), 7.88 (d, 2H), 7.90 (br.d, 1NH), 8.65 (d, 1NH), 9.1 ( br.S, OH).

B. Synthesis of N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((IR, 2R) -2- (hodriximethyl) cyclopropyl ) buta-1,3-diinyl) benzamide (1-2) Acetate of (E) -4-hodrixibut-2-enyl (2) To a solution of (E) -but-2-en-l, 4-diol 1 (264 g (3.0 mol) in THF 1.5 L, sodium hydride (120 g, 3.0 mol) was added in portions at -20 ° C. In the addition, the mixture was kept stirring at -20 ° C for 30 mins, then acetyl chloride (235.5g, 3 mol) was added dropwise, the mixture was allowed to warm to room temperature and kept stirring at room temperature for another 3 hours The mixture was filtered and the residue was washed with THF The combined organic layer was dried and concentrated to give the crude product 2 which was purified by column of silica gel (PE: EA = 5: 1- 2: 1) to give 2. 210 g as a colorless oil Yield: 54% RMN1H: CP-0005065-043 (CDC13, 400 M HZ) d: 5.85 (m, 1H), 5.62 (m, 1H), 4.67 (t, J = 6.2 Hz, 2H), 4.26 (t, J = 6.0 Hz, 2H), 2.10 (s, lH), 2.06 (s, 3H).

(E) -4-Oxobut-2-enyl acetate (3) To a suspension of manganese dioxide (active, 1305g, 15mol) in 2.5L of dichloromethane, (E) -4-hydroxybutyl-2-enyl acetate 2 (195g) was added in portions. The mixture was kept stirring at room temperature for 48 hours. The mixture was filtered and the residue was washed with dichloromethane. The combined organic layer was dried and concentrated to give the crude product 3 which was purified by silica gel column (PE: EA = 10: 1-5: 1) to give 3.30g as a colorless oil. Performance: 64%. RM 1H: CP-0005065-044 (CDC13, 400 M HZ) d: 10.01 (d, J = 6.4Hz, 1H), 6.52 (m, 1H), 6.10 (m, 1H), 5.08 (m, 2H), 2.10 (s, 3H).

Acetate of (E) -4,4-diethoxybutyl-2-ene (4) To a solution of (E) -4-oxobut-2-enyl 3 acetate (96.0 g, 0.75 mol) and triethoxymethane (133.2 g, 0.9 mol) in 500 ml of ethanol, ammonium nitrate (3.0 g, 0.038) was added. mol), the mixture was kept stirring at room temperature for 15 hours. The mixture was diluted with 800 ml of EtOAc and washed with saturated sodium bicarbonate. The aqueous layer was extracted again with EtOAc (300ml x 2). The combined organic layers were dried and concentrated to give the crude product 4 140 g as a red oil which was used for the next step without further purification. (4R, 5R, E) -diisopropyl-2- (3-acetoxyprop-1-enyl) -1,3-dioxolane-4,5-dicarboxylate (5) enyl 4 (60.6g, 0.3 mol) and (2R, 3R) -diisopropyl 2,3-dihodrixisuccinate (77.2g, 0.33 mol) in 500 ml benzene, PPTS (3.8 g, 15 mmol) was added, the mixture was heated to 90 ° C to distill the ethanol for 15 hours. The mixture was cooled to room temperature and concentrated in vacuo. It was purified by silica gel with (PE: EA = 50: 1-30: 1), 38.5 g 5 were obtained as a colorless oil. Yield: 37.3%. GCMS: CP 0005065-070-2 (85% pure). (4R, 5R) -diisopropyl-2- ((15, 2R) -2- (acetoxymethyl) cyclopropyl) -1,3-dioxolane-4,5-dicarboxylate (6) To a solution of (4R, 5R, E) -diisopropyl-2- (3-acetoxyprop-1-enyl) -1,3-dioxolane-4,5-dicarboxylate 5 (34.4 g, 0.1 mol) in hexane (1.5L) ), diethyl zinc (1M in hexane, 500 mL) was added in portions under argon at -20 ° C. In the addition, diiodoethane was added dropwise below -20 ° C with strong stirring. The mixture was allowed to warm to room temperature and was kept stirring for another 8 hours. The reaction mixture was quenched with 800 ml of cold aqueous ammonium chloride, then extracted with ether (800 ml × 5). The combined organic layer was washed with aqueous sodium thiosulfate, water, brine, then dried and concentrated to give the crude product 6 which was purified by silica gel column (PE: EA = 30: 1-10: 1) to give 6.16 g as a colorless oil. Performance: 44.7%. NMRH: P05 (CDC13, 400 MHZ) d: 5.13 (m, 2H), 4.95 (d, J = 5.6 Hz, 1H), 4.67 (d, J = 3.6 ??, ??), 4.57 (d, J = 4.0 ??, ??), 4.07 (m, 1H), 3.88 (m, lH), 2.06 (s, 3H), 1.38 (s, 12H), 1.23 (m, lH), 0.83 (m, lH), 0.66 (m, lH).

Acetate ((IR, 2R) -2-formylcyclopropyl) methyl (7) A mixture of (4R, 5R) -diisopropyl-2- ((1S, 2R) -2- (acetoxymethyl) cyclopropyl) -1,3-dioxolane-4,5-dicarboxylate 6 (14.3 g, 40 mmol) in 140 ml of 80% acetic acid was heated to 80 ° C and kept stirring at this temperature for 2 hours. When the TLC showed little remaining 6, the mixture was added to 300 ml of saturated sodium bicarbonate dropwise, then extracted with dichloromethane (200 ml x 3). The combined organic layers were washed with water, dried with brine and concentrated to give the crude product 7 which was purified by silica gel column (PE: EA = 10: 1-5: 1) to give 7. 3.5 g. as a colorless oil. Performance: 62%. NMR1H: CP-0005065-072 (CDC13.400MHZ) d: 9.15 (s, 1H), 4.11 (m, 1H), 3.91 (m, lH), 2.08 (s, 3H), 2.10 (s, 3H), 1.88 (m, 2H), 1.39 (m, 1H), 1.12 (m, 1H).

Acetate ((IR, 2R) -2- (2,2-dibromovinyl) cyclopropyl) methyl (8) To a solution of carbon tetrabromide (13.9 g, 42 mmol) in dichloromethane 30 mL, a solution of triphenylphosphine (22.0 g, 84 mmol) in 50 mL of dichloromethane was added dropwise at -20 ° C under argon. The mixture was kept stirring at this temperature for half an hour, then cooled to -78 ° C. A solution of ((1R, 2R) -2-formylcyclopropyl) methyl acetate (3.00 g, 21 mmol) in 40 ml of dichloromethane was added dropwise, the temperature was maintained for another half hour. The mixture was allowed to warm to room temperature for 30 mins. The solvent was removed and purified by silica gel column (PE: EA = 100: 1 ~ 50: 1) to give 8 as a colorless oil 4.3g. Performance: 69%. NMR ^ CP-0005065-075 (CDC13, 400 MHZ) d: 5.85 (d, J = 8.8Hz, 1H), 3.98 (m, 2H), 2.09 (m, 3H), 1.61 (m, 1H), 1.35 ( m, 1H), 0.88 (m, 2H). 4- (((IR, 2R) -2- (Acetoxymethyl) cyclopropyl) buta-1,3-diinyl) -benzoic acid methyl ester (10) To a solution of ((IR, 2R) -2- (2, 2-dibromovinyl) cyclopropyl) methyl acetate (4.3g, 14.5mmol), Pd2dba2 (86.3mg, 0.15mmol), tri (4-methylphenyl) phosphine (204mg, 0.58mmol), triethylamine (4.35g, 43.5mmol) in DMF 100 mL, was treated with methyl 4-ethylbenzoate 9 (2.64g, llmmol) under argon. The mixture was kept stirring at room temperature for 5 hours. When the TLC showed little remaining compound 8, the reaction was diluted with EtOAc (300 mL) and washed with water (3x100 mL), the organic layer was dried and concentrated to give the crude product which was purified by column chromatography. of silica gel PE: EA = 50: 1 -30: 1 to give 10 2.0 g as a yellow solid. Yield: 46.5%, LCMS: CP-0005065-085-2 (ESI) m / z = 297 (M + 1) purity: 92.4% (214nm). 4- (((IR, 2R) -2- (Hodriximethyl) cyclopropyl) buta-1,3-diinyl) benzoic acid (11) Methyl 4- (((IR, 2R) -2- (acetoxymethyl) cyclopropyl) buta-1,3-diinyl) benzoate (1.92g, 6.5mmol) in THF (40 mL) was dissolved, then a solution was added of sodium hydroxide (2.60 g, 65 mmol) in lOmL of water. The mixture was kept stirring at room temperature for 8 hours. When the LCMS showed little remaining compound, the solvent it was removed under reduced pressure, the residue was diluted with water (50 mL), the pH adjusted to 4.0 (extracted with ethyl acetate (4x50 mL), the organic layer was dried and concentrated to give the crude product 1.4 g. as a yellow solid that was used for the next step without further purification LCMS: CP-0005065-088-3 (ESI) m / z = 241 (+ l) purity: 89% (214nm) Yield: 89%. 2- (4- (((IR, 2R) -2- (Acetoxymethyl) cyclopropyl) buta-1,3-diinyl) -benzamido) -3- (tert-butoxycarbonylamino) -3-methylbutanoate of (S) -methyl ( 12) a solution of 4- (((IR, 2R) -2- (hodrix methyl) cyclopropyl) buta-1,3-diinyl) benzoic acid 11 (1.20g, 5.0 mmol), HATU (2.34 g, 6 mmol) in DMF 50mL was treated with (S) -methyl 2-amino-3- (tert-butoxycarbonylamino) -3-methylbutanoate (1.48g, 6. OmmoL) and DIPEA (3.58g, 20mmoL). The mixture was kept stirring at room temperature for 5 hours. When the LCMS showed little remaining compound, the reaction was diluted with EtOAc (100 mL), washed with 5% lithium chloride (3x50 mL), the organic layer was dried and concentrated to give 12 as a yellow oil. Purification by column chromatography on silica gel PE: EA = 2: 1 gave 2.Og 12 as a colorless oil, yield: 70%, LCMS: CP-0005065-091-3 (ESI) m / z = 469 (M + 1) purity: 95% (214nm). 3-Amino-2- (4- (((IR, 2R) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate hydrochloride of (S) -methyl (13 ) 2- (4- (((IR, 2R) -2- (acetoxymethyl) -cyclopropyl) buta-1,3-diinyl) benzamido) -3- (tert-butoxycarbonylamino) -3-methylbutanoate (S) was dissolved ) -methyl 12 (1.87g, 4.0mmoL) in methanol (50mL), was treated with dry HClg for lOmins. When the LCMS showed little remaining compound 12, the flow of HC1 was stopped. The solvent was removed under reduced pressure, and gave 13, 1.45 g as a yellow solid. Yield: 91%, LCMS: CP-0005065-096-4-LCMSA019 (ESI) m / z = 369 (M + 1) purity: 98% (214nm). RMN1 !! : C -0005065-096-4 (DMSO-dff, 400 Hz) 5: 9.04 (d, J = 6.8Hz, 1H), 8.36 (s, 3H), 7.98 (d, J = 6.4Hz, 2H), 7.64 (d, J = 6.8Hz, 2H), 4.89 (d, J = 6.8Hz, 1H), 3.73 (s, 3H), 3.44 (m, 1H), 3.22 (m, 1H), 1.48 (m, 2H) , 1.40 (s, 6H), 0.94 (m, 2H). 3-Amino-2- (4- (((IR, 2R) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate hydrochloride of (S) -methyl 13 ( 1.25g, 3.1mmol), with isopropanol (10mL) and 50% aqueous hodrixylamine (4.1mL, 6l.7mmol, 20 equivalents) until it is predominantly completed by LCMS.

The crude material was purified by reverse phase HPLC (gradient of 0-30% acetonitrile in water, each containing 0.1% TFA, for 120 minutes) and the desired fractions were mixed and lyophilized to give N- ((S ) -3-amino-l- (hydroxy-amino) -3-methyl-l-oxobutan-2-yl) -4- (((IR, 2R) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) -benzamide as its trifluoroacetate salt (white solid, 647mg, 1.3mmol, 43%). Mass spectrometry data: expected (M + l): 370.4, observed: 370.2. Proton MRI (400MHz, dmso-d6): 11.20 (s, 1H), 9.43 (br s, 1H), 8.55 (d, 1H, J = 9.6Hz), 8.00 (br s, 3H), 7.89 (dd, 2H, J = 1.8, 6.6Hz), 7.60 (dd, 2H, J = 2.0, 6.8Hz), 8.66 (d, 1H, J = 9.2Hz), 3.39 (dd, 1H, J = 4.8, 11.6Hz), 3.22 (dd, 1H, J = 5.8, 11.4Hz), 1.39 - 1.46 (m, 2H), 1.30 (s, 3H), 1.25 (s, 3H), 0.84 - 0.93 (m, 2?) C. Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2S) -2- (hodriximethyl) cyclopropyl buta-1, 3-diinyl) benzamide (1-3) (4S, 5S, E) -diisopropyl-2- (3-acetoxyprop-1-enyl) -1, 3-dioxo-lane-4,5-dicarboxylate (5) To a solution of (E) -4,4-diethoxybut-2-enyl 4 acetate (75 g, 0.37 mol) and diisopropyl (-) - (S, S) -tartrate (86.8 g, 0.3 mol) in 800 ml of benzene, PPTS (4.66 g, 0.0185 mol) was added, the mixture was heated to 9 ° C to distill the ethanol for 15 hours. The mixture was cooled to room temperature and concentrated in vacuo. It was purified by distillation to give compound 5 (50.0 g, 39%) as a colorless oil. NMR ¾: (CDC13, 400 MHz) d: 6.04-6.01 (m, 1H), 5.81-5.80 (m, 1H), 5.57 (d, J = 8Hz, 1H), 5.07-5.01 (m, 2H), 4.65 (d, J = 4 Hz, 1H), 4.57 (d, J = 4 Hz, 1H), 4.54-4.53 (m, 2H), 1.99 (s, 3H), 1.23-1.19 (m; 12H). (4S, 5S) -diisopropyl-2- ((IR, 2S) -2- (acetoxymethyl) cyclopropyl) -1,3-dioxolan-4,5-dicarboxylate (6) acetoxypropyl-l-enyl) -1,3-dioxolane-4,5-dicarboxylate 5 (40g, 0.12mol) in hexane (1.0L), diethyl zinc (1M in hexane, 1.16L) was added in portions under argon a -20 ° C. In the addition, diiodomethane (623g, 2.40mol) was added dropwise below -20 ° C with strong stirring. The mixture was allowed to warm to room temperature and was kept stirring for another 8 hours. The reaction mixture was quenched with 800 ml of cold aqueous ammonium chloride, then extracted with ether (800 mL x 5). The combined organic layer was washed with aqueous sodium thiosulfate, water, brine, then dried and concentrated to give crude compound 6 which was purified by column of silica gel with ethyl acetate in petroleum ether (3% -10%). % v / v) to give compound 6 (20.0 g, 50%) as a colorless oil. NMR ?: (CDC13.400 MHz) d: 5.15-5.08 (m, 2H), 4.95 (d, J "= 5.6 ??, ??), 4.67 (d, J = 3.6 ??, ??), 4.57 (d, J = 4.0 ??, ??), 4.07-4.04 (m, 1H), 3.91-3.86 (m, 1H), 2.06 (s, 3H), 1.40-1.37 (m, 1H), 1.31-1.28 (m, 12H), 1.24-1.22 (m, lH), 0.85-0.82 (m, 1H), 0.68-0.63 (m, 1H). ((1S, 2S) -2-formylcyclopropyl) methyl acetate (7) (acetoxymethyl) cyclopropyl) -1,3-dioxolane-4,5-dicarboxylate 6 (20. Og, 55.8mmol) in 200ml of 80% acetic acid was heated to 80 ° C and kept stirring at this temperature for 2 hours. When the TLC showed little remaining 6, the mixture was diluted with water (150mL), extracted with ethyl acetate (200ml x 3). The combined organic layer was washed with saturated sodium bicarbonate, water, brine, then dried and concentrated to give crude compound 7 which was purified by column of silica gel with ethyl acetate in petroleum ether (8% -20% v / v) to give compound 7 (3.0, 38) as a colorless oil. NMR ??: (CDC13, 400 MHz) d: 9.15 (d, J = 4.8, 1H), 4.14-4.09 (m, 1H), 3.95-3.90 (m, 1H), 2.08 (s, 3H), 1.92- 1.88 (m, 2H), 1.38-1.36 (m, 1H), 1.12-1.09 (ra, 1H).

Acetate ((1S, 25) -2- (2,2-dibromovinyl) cyclopropyl) methyl To a solution of carbon tetrabromide 13.9g, 42.0 mmol) in dichloromethane (30mL), a solution of triphenylphosphine (22.0g, 84.0mmol) in 50mL of dichloromethane was added dropwise at -20 ° C under argon. The mixture was kept stirring at this temperature for half an hour, then cooled to -78 ° C. A solution of ((1S, 2S) -2-formylcyclopropyl) methyl acetate (3.0g, 21.0mmol) in 40mL of dichloromethane was added dropwise, the temperature was maintained for another half hour. The mixture was allowed to warm to room temperature for 30 mins. The solvent was removed under vacuum and the residue was purified by column of silica gel with petroleum ether to give compound 8 (3.20g, 51) as a colorless oil. It was confirmed by GC-MS. 4- (((1S, 2S) -2- (Acetoxymethyl) cyclopropyl) buta-1,3-diinyl) -benzoic acid methyl ester (10) To a solution of ((1S, 2S) -2 - (2,2-dibromovinyl) cyclopropyl) methyl acetate (3.2g, 10.85mmol), Pd2dba2 (62.4mg, O.llmmol), tri (4-methylphenyl) phosphine (153mg, 0.43mmol), triethylamine (1.44g, 14.25mmol) in DMF. (50mL), treated with methyl-ethylbenzoate 9 (2.43g, 15.2mmol) under argon. The mixture was kept stirring at room temperature overnight. The reaction was diluted with EtOAc (300 mL), washed with water (3 x 100 mL), the combined organic layer was dried and concentrated to give crude compound which was purified by silica gel column chromatography with ethyl acetate in Petroleum ether (3% -10% v / v) to give compound 10 (1.4g, 46.5%) as a yellow solid. LC-MS: 297 [M + H] +. 4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzoic acid (11) methyl cyclopropyl) buta-l, 3-diinyl) benzoate 10 (1.4 g) in THF (30 mL) was added a solution of sodium hydroxide (75 mg, 18.9 mmol) in water (10 mL). The mixture was kept stirring at room temperature overnight. After this time, the solvent was removed under reduced pressure, the residue was diluted with water (50mL), the pH was adjusted to 4.0 with 1M HC1, then extracted with EtOAc (4 x 50mL), the organic layer was dried and Concentrate to give compound 11 (1.05g, 93%) as a yellow solid which was used for the next step without further purification. LC-MS-.241 [? +?] +. ' 3- (tert-Butoxycarbonylamino) -2- (4- (((1S, 2S) -2- (hydroxymethyl) -cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (S) -methyl (12) To a solution of 4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) enzoic acid 11 (1.05g, 4.38 mmol), HATU (1.83g, 4.81raraol) in DMF (25 mL) was added 2-amino-3- (tert-butoxycarbonylamino) -3-methylbutanoate (S) -methyl (1.076 g, 4.38 mmol) and DIPEA (1.69 g, 13.1 mmol). The mixture was kept stirring at room temperature for 5 hours. After this time, the reaction was diluted with water (20mL), then extracted with ethyl acetate (60mL x 3). The combined organic layers were washed with water and brine, then dried and concentrated to give Compound 12 as a yellow oil. (1.35g, 65%) LC-MS: 469 [M + H] +. 3-Amino-2- (4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate hydrochloride (S) -methyl (13) A solution of 3- (tert-butoxycarbonylamino) -2- (4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate of (S) - methyl 12 (1.3g, 2.78mmol) in anol (20mL) was bound to an HC1 apparatus. The reaction was then stirred at room temperature until TLC indicated the total consumption of the starting material. After that, the solution was concentrated under reduced pressure to give 13 as a yellow solid (1.1 g, 98%). LC-MS: 369 [M + H] + RMN-1 !! : (DMSO-de, 400 MHz) 5: 9.04 (d, J = 6.8Hz, 1H) 8.38 (s, 3H), 7.98 (d, J = 6.8Hz, 2H), 7.64 (d, J = 6.4Hz, 2H) 4.88 (d, J = 6.8Hz, 1H), 3.72 (s, 3H), 3.45-3.42 (m, 1H), 3.28 3.24 (m, 1H), 1.49-1.46 (m, 2H), 1.40 (s) , 6H), 0.95-0.90 (m, 2H). 3-Amino-2- (4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate hydrochloride of (S) -methyl 13 ( 1.15g, 2.8mmol) with isopropanol (lOmL) and 50% aqueous hydroxylamine (3.8mL, 56.8mmol, 20 equivalents) until complete predominantly by LCMS.

The raw material. was purified by reverse phase HPLC (gradient of 0-30% acetonitrile in water, each containing 0.1% TFA, for 120 minutes) and the desired fractions were mixed and lyophilized to give N- ((S) -3 -amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide as its Trifluoroacetate salt (white solid, 446mg, 0.92mmol, 33%). Mass spectrometry data: expected (M + l): 370.4, observed: 370.2. RM of Proton (400MHz, dmso-d6): 11.20 (s, 1H), 9.22 (br s, 1H), 8.55 (d, 1H, J = 9.6Hz), 7.99 (br s, 3H), 7.89 (dd, 2H, J = 2.0, 6.8Hz), 7.61 (dd, 2H, J = 1.6, 6.8Hz), 4.66 (d, 1H, J = 9.6Hz), 3.40 (dd, 1H, J = 5.0, 11.4Hz), 3.22 ( dd, 1H, J = 5.8, 11.8Hz), 1.38 - 1.46 (m, 2H), 1.30 (s, 3H), 1.25 (s, 1H), 0.84 - 0.93 (m, 2H).

D. Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 '- (((1, 2-trans) -2- ( hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-carboxamide (1-5) To a round bottom flask was added methyl 4'-bromobiphenyl-4-carboxylate (2.0 g, 6.87 mmol, 1.0 equiv) followed by a solution of racemic 2-trans-ethynylcyclopropyl methanol (0.991 g, 10.30 mmol, 1.5 equiv. ) dissolved in THF (20 mL). To this was added bis-triphenylphosphine of palladium dichloride (241 mg, 0.343 mmol, 0.05 equiv), copper iodide (I) (131 mg, 0.687 mmol, 0.1 equiv), then triethylamine (6.87 mL, 49.3 mmol, 7.18 equiv). The reaction was stirred at 75 ° C for ~ 2 h. The reaction was cooled to room temperature, the solids were completely filtered, rinsed with THF. The solution was placed in the freezer for 5 days. It was concentrated to dryness. DCM was added and a precipitate formed. The precipitate is filtered, rinsed with minimal DCM. TLC shows that ppt contains the majority of the product (~ 1.5g of solid). Triturate with 10% EtOAc in hexanes (5 mL), extract the solvent with pipette, TLC shows that the solid contains the product and the liquid removed the starting material-bromide and some baseline impurities. Additional 10% EtOAc in hexanes (5 mL) was added and stirred for 30 minutes, filtered, rinsed with 10% EtOAc in hexanes (5 mL), and the solid dried. 900 mg of crude 41 - (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) ethynyl) bipheni-4-methylcarboxylate was produced. RM - ^ (DMS0-d6): d 0.82-0.85 (m, 2H), 1.35-1.43 (m, 2H), 3.23-3.29 (m, 1H), 3.40-3.42 (m, 1H), 3.84 (s, 3H), 4.65-4.67 (t, 1H), 7.43-7.45 (d, 2H), 7.67-7.70 (d, 2H), 7.80-7.82 (d, 2H), 7.99-8.01 (d, 2H).

Methyl 41 - (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-carboxylate (900 mgs, 2.94 mmol, 1.0 equiv) was dissolved in methanol (5 mL), DMF (2 mL), and THF (5 mL). At room temperature, NaOH was added 1. 0M (4.41 mL, 4.407 mmol, 1.5 equiv). The reaction was stirred for 4 days. The reaction was concentrated, to remove eOH and THF, acidified to pH ~ 3 with 6N HC1 (~5 mL). Extract with EtOAc (3x50 mL), combine the organic layers, wash with saturated NaCl, dry (MgSO 4), filter, concentrate. 890 mg of 41 - (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-carboxylic acid was produced. R N-¾ (DMSO-d6): d 0.79-0.88 (m, 2H), 1.34-1.44 (m, 2H), 3.22-3.26 (m, 1H), 3.40-3.44 (m, 1H), 7.42-7.44 (d, 2H), 7.S6-7.68 (d, 2H), 7.76-7.78 (d, 2H), 7.97-7.99 (d, 2H), 8.10 (s, 1H). 2-Amino-3- (tert-butoxycarbonylamino) -3-methylbutanoate of (S) -methyl (150 mg, 0.609 mmol, 1.0 equiv) and 4 '- (((1-2) rans) -2- were dissolved. (hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-carboxylic acid (178 mg, 0.609 mmol, 1.0 equiv) in DMF (2 mL). To this was added DIPEA (0.266 mL, 1.523 mmol, 2.5 equiv) then HATU (278 mg, 0.731 mmol, 1.2 equiv). This was stirred at room temperature for ~ 48 h. The reaction was dissolved between 1M citric acid and ethyl acetate. The organics were washed with semi-saturated sodium chloride, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. 370 mg of 3- (tert-butoxycarbonylamino) -2- (4 '- (((S, 2S) -2- (hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-ylcarboxamido) -3-methylbutanoate were produced (S) -methyl crude. LCMS M + l expected = 521.3, observed = 521.3.

(S) -methyl-3- (tert-butoxycarbonylamino) -2- (4 '- (((1, 2-trans) -2- (hydroxymethyl) -cyclopropyl) ethynyl) biphenyl-4-ylcarboxamido) -3 was dissolved. -methyl-butanoate (crude) in methanol (1 mL). At room temperature, 4.0M HC1 in dioxane (1.283 mL, 5.13 mmol, 8.43 equiv) was added. The reaction was complete after 90 minutes (HPLC). The reaction was completed in an evaporator challenge at 0 ° C. To this was added IPA (1 mL) then hydroxylamine solution (0.804 mL, 12.18 mmol, 20 equiv). The flask was placed at 4 ° C for ~ 120h. The reaction was concentrated, (maintaining the reaction at 0 ° C) to a gummy mass. To this was added water (3 mL) and ACN (0.5 mL). It was acidified while it is 0 ° C using TFA (3 mL). Additional water (1 mL) and ACN (1 mL) were added. Purified by RP HPLC (1"column, 25mL / min, '0.1% TFA in water / ACN, 10% B equil) was loaded onto a 1" column (10mL / min, 5% B) using syringe filter (2X6.5 mL). It was increased to 25mL / min for 1 minute. 10% B for 15 minutes, then 10-70% B for 80 minutes, the product eluted at 41-48 minutes. The desired fractions were combined, frozen and placed in ice. 105 mg of N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -41 - (((1, 2-trans) -2- ( hydroxymethyl) cyclopropyl) ethynyl) biphenyl-4-carboxamide (1-5), TFA, LCMS M + l expected = 422.2, observed = 422.2. RMN-1 !. (DMSO-d6): d 0.80-0.88 (m, 2H), 1.27 (s, 3H), 1.31 (s, 3H), 1.36-1.44 (m, 2H), 3.21-3.29 (m, 1H), 3.40- 3.45 (m, 1H), 4.65-4.71 (m, 2H), 7.44-7.46 (d, 2H), 7.68-7.71 (d, 2H), 7.79-7.81 (d, 2H), 7.97-7.99 (d, 2H) ), 8.46-8.48 (d, 1H), 9.22 (s, 1H), 11.21 (br, 1H).

AND_. Synthesis of N- ((S) -1- (hydroxyamino) -3-methyl-3- (methylamino) -l-oxobutan-2-yl) -4 - (((1,2-trans) -2 - (hydroxymethyl) ) cyclopropyl) buta-l, 3-diinyl) benzamide (1-6) and N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl ) -4- (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide (1-7) 2) NaCNBH3 / AcOH / THF I-6, R = CH3 1-7, R = CH2CH3 Into a 250mL round bottom flask were added N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- (((1, 2-trans) ) -2- (hydroxymethyl) cyclopropyl) -buta-1, 3-diinyl) benzamide (1-1) (1.2g, 3.25 mmol), triethylamine (2 mL, 14.35 mmol), and formaldehyde (0.2 g, 6.66 mmol) in DMF (20 mL) to give a yellow solution. The reaction mixture was stirred for 3 h. The excess formaldehyde was quenched with n-butylamine (2 ml). Sodium cyanoborohydride (570mg, 14.3mmol) and acetic acid (4ml) were added at room temperature at 0 ° C. The reaction mixture was verified by LCMS. After removing the solvents, the product was purified by HPLC to give 440mg of N- ((S) -1- (hydroxyamino) -3-methyl-3- (methylamino) -1-oxobutan-2-yl) -4- (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide, 1-6. LC-MS (M + l) 384; Chemical formula: C2iH25 304, Exact Mass: 383.18. NMR-XH (DMSO-d6), TFA salt: d 0.88 (m, 2H) 1.30 (s, 3H), 1.35 (s, 3H), 1.44 (m, 1H), 2.51 (s, 'H), 3.39 (m, 1H), 3.41 (m, 1H), 4.7 (br.d, 1H), 4.84 (d, 1H), 7.58 (d, 2H), 7.91 (d, 2H), 8.51 (br.m, 1 H) 8.64 (d, 1NH), 9.23 (S, NH). 11.17 (s, OH).

Into a 250mL round-bottomed flask were added N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- (((1,2-trans) ) -2- (hydroxymethyl) cyclopropyl) -buta-1,3-diinyl) benzamide (1-1) (lg, 2.71mmol), triethylamine (0.377 ml, 2.71 mmol), and acetaldehyde (0.119 g, 2.71 mmol) in DMF (20 ml) to give a yellow solution. The reaction mixture was stirred for 3h. At room temperature sodium cyanoborohydride (570mg, 14.3mmol) and acetic acid (2ml) were added, the reaction mixture was purified by LCMS. The reaction mixture was concentrated and the product was purified by HPLC to give 642mg of N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-1-oxobutan-2-yl) -4- (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide, 1-7. LC-MS (M + l) 398; Chemical formula: C22H27 304, Exact mass: 397.20. NMR-XH (DMSO-d6), TFA salt: d 0.85 (m, 2H), 1.14 (t, 3H), 1.29 (s, 3H), 1.39 (s, 3H), 1.44 (m, 1H), 2.96 (br.m, 2H) 3.24 (m, 2H) 3. 39 (m, 2H), 4.64 (br.d, 1H), 4.85 (d, 1H), 7.60 (d, 2H), 7.92 (d, 2H), 8.59 (br.m, 1NH) 8.45 (brt, 1NH), 9.23 (S, NH), 11.15 (s, OH).

F. Synthesis of N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) - 2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide (1-8) Into a 250 mL round bottom flask (t = g) was added N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (( (1, 2-trans) -2- (hydroxymethyl) cyclopropyl) uta-1,3-diinyl) benzamide (lg, 2.71mmol), and triethylamine (0.377ml, 2.71mmol) in DMF (20 ml) to give a solution yellow. 2- (tert-Butyldimethylsilyloxy) acetaldehyde (0.494g, 2.83mmol) was added. The reaction mixture was stirred for 18 h, concentrated to dryness. The residue was dissolved in THF (15ml) and acetic acid (2ml) and sodium cyanoborohydride (600mg, 15mmol) were added at room temperature. The reaction mixture was verified by LCMS. After stirring at room temperature for 2 h, TFA (5 ml) was added and stirred for an additional 3 hours. The end of the reaction was confirmed by LCMS and the reaction was concentrated and the product was purified by HPLC to give 370mg of N- ((s) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl- l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diin l) benzamide, I- 8. LC-MS (M + l 414; Chemical formula: C22H27 3O5, Exact Mass: 413.20; NMR-1H (DMSO-d6), TFA salt: d 0.88 (m, 2H) 1.30 (s, 3H), 1.35 (s, 3H), 1.44 (m, 1H), 3.22 (m, 2H), 3.39 ( m, 2H), 4.64 (br.d, 1H), 4.90 (d, 1H), 5.25 (br.S, 1H) 7.60 (d, 2H), 7.91 (d, 2H), 8.59 (br.m, 1NH ) 8.71 (d, 1NH), 9.26 (S, NH). 11.19 (s, OH).

G. Synthesis of N- ((S) -3- (dimethylamino) -1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide (1-9) N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ((2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl was dissolved ) enzyme (300 mg, 0.81 mmol) in N, N-dimethylformamide (10 mL) and treated with paraformaldehyde (732 mg, 8.1 mmol, lOeq) and W / N-diisopropylethylamine (0.56 mL, 3.3 mmol, 4 eq) at room temperature for 16 h. hours. Trifluoroacetic acid (1.3mL, 16.2mmol, 20eq) and sodium cyanoborohydride (lOlmg, 1.6mmol, 2 eq) were introduced.

The crude material was purified by reverse phase HPLC and the desired fractions were mixed and lyophilized to give N- ((S) -3- (dimethylamino) -1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl. ) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide as its trifluoroacetate salt (white solid, 2.9mg, 5.3umol, 0.7%). Mass spectrometry data: expected (M + l): 398.5, observed: 398.2. Proton NMR (400MHz, dmso-d6): 11.16 (s, 1H), 9.25 (br s, 1H), 9.05 (br s, 1H), 8.68 (d, 1H, J = 10.0Hz), 7.94 (dd, 2H, J = 1.6, 8.0HZ), 7.59 (dd, 2H, J = 1.8, 8.2Hz), 5.06 (d, 1H, J = 9.6Hz), 3.4 (peak darkened by water), 3.21 (dd, 1H, J = 5.2, 11.2Hz), 2.75 (d, 3H, J = 5.6Hz), 2.73 (d, 3H, J = 5.2Hz), 1.48 (s, 3H), 1.39 - 1.45 (m, 2H), 1.28 ( s, 3H), 0.85-0.91 (m, 2H).

H. Synthesis of N- ((S) -3-hydroxy-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1, 2-trans) -2- (hydroxymethyl) ) cyclopropyl) buta-1,3-diinyl) benzamide (1-10) To a mixed solution of 4- ((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzoic acid (750 mg, 3.12 mmol), HATU (570 mg, 1.5 mmol) and DIEA (3.4 ml in excess) in DMF (30 ml) at room temperature was added 2-amino-3-hydroxy-3-methylbutanoate hydrochloride (S) -methyl (402 mg, 1.5 mmol). The reaction mixture was stirred at room temperature for 1 hour followed by dilution with water (50 ml). The solution was extracted with ethyl acetate (100 ml x 3) and brine (20 ml). The organic layer was dried over MgSO4 and evaporated. The resulting product (1.3g) was dissolved in IPA (20mL) and treated with NH2OH (10mL, in excess) and stirred at room temperature for 3 days. The excess solvent was removed and the crude product was purified on a reverse phase HPLC to give 178 mg of N- ((S) -3-hydroxy-1- (hydroxyamino) -3-methyl-1-oxobutan-2 il) -4- (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1, 3-diinyl) benzamide 1-10; LC-MS (M + l) 371; Chemical formula: C2oH22N205 Exact Mass: 370.15. NMR-Hl (DMSO-d6),: d 0.88 (m, 2H) l.ll (s, 3H), 1.16 (s, 3H), 1.44 (m, 1H), 3.20 (m, 1H), 3.40 (m , 2H), 4.34 (d, 1H), 4.84 (d, 1H), 7.58 (d, 2H), 7.84 (d, 2H), 7.6 (d, 1NH), 8.88 (br.s, NH). 10.57 (s, OH).

I. Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2R) -2- (hydroxymethyl) - 2- methylcyclopropyl) buta-1,3-diinyl) benzamide (1-11) ACHL02-112-Intermediate-A compound 3- (2, 2-dimethyl-l, 3-dioxolan-4-yl) -2-methylacrylate (S, E) -ethyl (3) PM AC reagent. mmol 9, mL Compound 1 262 30 1 49.6 13 0g Nal04 213 89 1.2 59.5 12 7 g Compound 2 238 22 2 99.2 23 0 g K2C03 138 15 2.1 103.6 14 3 g NaHCO3 (5% ac) 60 mL shaken solution of D-mannitol-diacetone (13.0 g, 49.6 mmol) in sodium bicarbonate (5% ac, 60 ml), an aqueous solution of sodium periodate (12.74 g, 59.5 mmol) was added dropwise at room temperature. In the addition, the mixture was kept stirring at room temperature for 2 hours. Then ethyl 2- (diethoxyphosphoryl) was added 2 (23.0 g, 99.2 mmol) followed by potassium carbonate (14.3 g, 103.6 mmol), the mixture was kept stirring at room temperature for another 120 hours. The mixture was added to 500 ml of water, extracted with EtOAc (3ÓÓmLx3). The combined organic layer was dried over anhydrous sodium sulfate, concentrated and then chromatographed with PE: EA (30 μl) to give the desired compound 3 5.5 g as a colorless oil, yield: 51.8%.

(S, E) -3- (2, 2-dimethyl-l, 3-dioxolan-4-yl) -2-methylprop-2-en-l-ol (4) To a solution of (S, E) -ethyl 3 (2, 2-dimethyl-l, 3-dioxolan-4-yl) -2-methylacrylate (5.35 g, 5 mmol) in THF 150 ml, was added a solution of DIBAL-H (50 mL, 50 mmol) dropwise under argon at 0 ° C. In the addition, the mixture was kept stirring at 0 ° C for 3 hours. When the TLC showed little remaining compound 3, the reaction was quenched with 30 mL of aqueous ammonia and stirred with 300 mL of dichloromethane for another hour. The mixture was filtered and the residue was washed with THF, the combined organic layer was dried and concentrated to give the crude product 4 which was purified by column of silica gel with PE: EA = 10: 1-5: 1, 3.0 g Compound 4 as a colorless oil was obtained, Yield: 70%.

Acetate of (S, E) -3- (2, 2-dimethyl-l, 3-dioxolan-4-yl) -2-methylallyl (5) To a solution of (S, E) -3- (2,2-dimethyl-1,3-dioxolan-4-yl) -2-methylprop-2-en-l-ol 4 (2.92 g, 17 mmol) in THF 100 mL, triethylamine (4.3 g, 42.5 mmol) was added. Acetyl chloride (2.7 g, 34 mmol) was then added dropwise under argon at 0 ° C. In the addition, the mixture was kept stirring at 0 ° C for 3 hours. When the TLC showed little Compound 4 remaining, the reaction was poured into 150 ml of water. It was extracted with EtOAc (50mLx3). The combined organic layer was dried over anhydrous sodium sulfate, concentrated and then chromatographed with PE: EA (10 μl) to give the desired compound 5. 3.4 g as a colorless oil, yield: 89%.

Acetate ((trans) -2- ((S) -2, 2-dimethyl-l, 3-dioxolan-4-yl) -1-methylcyclopropyl) methyl (6) a solution of (S, E) -3- (2, 2-dimethyl-1,3-dioxolan-4-yl) -2-methylallyl acetate (3.2 g, 15 mmol) in hexane 200 mL, diethyl ether was added. zinc (1M in hexane, 75 mL) in portions under argon at -20 ° C. After the addition, diiodomethane (40 g, 150 mmol) was added dropwise below -20 ° C with vigorous stirring. The mixture was kept stirring below minus -20 ° C for another 8 hours and allowed to warm to room temperature and kept stirring for another 8 hours. The reaction mixture was quenched with 100 ml of cold aqueous NH4C1, then extracted with ether (100 ml x 5). The combined organic layer was washed with aqueous sodium thiosulfate, water, brine and then dried and concentrated to give the crude product 6 which was purified by silica gel column (PE: EA = 30: 1-20: 1) to give 6.0.0.0 g as a colorless oil. Yield: 88%.

Acetate ((trans) -2- ((S) -1,2-dihydroxyethyl) -1-methylcyclopropyl) methyl (7) A mixture of ((trans) -2- ((S) -2,2-dimethyl-1,3-dioxolan-4-yl) -1-methylcyclopropyl) methyl acetate (2.97 g, 13 mmol) in 30 ml of 80% acetic acid was kept stirring at room temperature for 15 hours. Then the TLC showed few remaining 6, the mixture was added to 300 ml of saturated aqueous sodium bicarbonate drop by drop. It was then extracted with dichloromethane (200 ml x 3). The combined organic layer was washed with water, dried with brine and concentrated to give crude product 7 which was purified by silica gel column (PE: EA = 3: 1-1: 1) to give Compound 7, 1.5 g as a colorless oil. Performance: 62%. ((Trans) -2-formyl-l-methylcyclopropyl) methyl acetate (8) To a stirred solution of ((trans) -2- ((S) -1,2-dihydroxyethyl) -1-methylcyclopropyl) methyl acetate 7 (1.41 g, 7.5 mmol) in sodium bicarbonate (5% ac) 20 ml , a saturated solution of sodium periodate (1.93 g, 9 mmol) was added dropwise at room temperature. In the addition, the mixture was kept stirring at room temperature for 2 hours. When the TLC showed little remaining 7, the mixture was added to 50 ml of water, extracted with EtOAc (100mLx3). The combined organic layer was dried over anhydrous sodium sulfate, concentrated and then chromatographed with PE: EA (20/1) to give the desired compound 8. 1.0 g as a colorless oil, yield: 85%. ((trans) -2-ethynyl-l-methylcyclopropyl) methanol (9) To a solution of ((trans) -2-formyl-1-methylcyclopropyl) methyl acetate 8 (780 mg, 5 mmol) in 15 mL methanol, was added Bestmann's reagent (1.44 g, 7.5 mmol) and potassium carbonate ( 2.07 g, 15 mmol), the mixture was kept stirring at room temperature for 5 hours. When the TLC showed little remaining 8, the reaction mixture was diluted with water (20 mL). Extract with ether (20mLx3), the combined organic layer was dried and concentrated to give the crude product 9, which was purified by column on silica gel (PE: Et2O = 10: 1-5: 1) to give 9 as a colorless oil 400 mg. Performance: 73%. 4- (((trans) -2- (hydroxymethyl) -2-methylcyclopropyl) buta-1,3-diinyl) methyl benzoate (11) To a solution of ((trans) -2-ethynyl-l-methylcyclopropyl) methanol 9 (400 mg, 3.6 mmol) / Pd (PPh 3) 2 Cl 2 (126 mg, 0.18 mmol), Cul (68.4 mg, 0.36 mmol), DIPEA (1.09g, 10.8 mmol) in THF 20 mL, was treated with methyl 4 - (bromoetinyl) benzoate 10 (870 mg, 3.6 mmol) under argon. The mixture was kept stirring at room temperature for 5 hours. When the TLC showed little remaining compound 9, the solvent was removed under reduced pressure, the residue was diluted with water (50 mL), extracted with dichloromethane (3x50 mL), the organic layer was dried and concentrated to give the crude product. as a red oil which was purified by column chromatography on silica gel PE: EA = 20: 1-10: 1 to give 11,430 mg as a yellow solid. Performance: 40%. 4- (((1S, 2R) -2- (Hydroxymethyl) -2-methylcyclopropyl) buta-1,3-diinyl) benzoic acid (ACHL02- 112-intermediate-A) To a stirred solution of methyl 4- (((trans) -2- (hydroxymethyl) -2-methylcyclopropyl) buta-l, 3-diinyl) benzoate 11 (400 mg, 1.5 mmol) in THF 16 ml, was added a solution of monohydrate lithium hydroxide (240 mg, 6 mmol) in 4 mL of water. The addition, the mixture was kept stirring at room temperature for 15 hours. When LCMS showed little remaining compound 11, the mixture was added to 50 ml of water. The pH was adjusted to 3.0 with 1M HCl at 0 ° C, extracted with EtOAc (100mLx3). The combined organic layer was dried over anhydrous sodium sulfate, concentrated to give the desired compound 370 mg as a yellow powder, yield: 92%. LCMS: CP-0004344-006-3-03665-LCMSA035 (ESI) m / z = 255 (M + 1). NMR- CP-0004344-006-3 (DMS0-dff, 400 Hz) d: 13.1 (s, 1H), 7.91 (d, J = 8.4 Hz, 2H), 7.62 (d, J = 8.4Hz, 2H), 4.75 (s, 1H), 3.22 (m, 2H), 1.59 (m, 1H), 1.18 (s, 3H), 1.08 (m, 1H), 0.66 (m, 1H). 2-amino-3- (tert-butoxycarbonylamino) -3-methylbutanoate of (S) -methyl (347 mg, 1,408 mmol, 1.0 equiv) and 4- (((1S, 2R) -2- (hydroxymethyl)) were dissolved. -2-methylcyclopropyl) buta-1,3-diinyl) benzoic acid (358 mg, 1,408 mmol, 1.0 equiv) in DMF (3 mL). To this was added DIPEA (0.615 mL, 3.52 mmol, 2.5 equiv) then HATU (642 mg, 1.689 mmol, 1.2 equiv). This was stirred at room temperature for -16 h. The reaction was partitioned between 1M citric acid and ethyl acetate. The organics were washed with semi-saturated sodium chloride, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. 912 mg of 3- (tert-butoxycarbonylamino) -2- (4 - (((S, 2R) -2- (hydroxymethyl) -2-methylcyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate were produced. of (S) -methyl crude. LCMS M + l expected = 483.2, observed = 483.2. dissolved (S) -methyl-3- (tert-butoxycarbonylamino) 2- (4- (((1S, 2R) -2- (hydroxymethyl) -2-methylcyclopropyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (crude) in methanol (1 mL).

At room temperature, HC1 4. OM in dioxane (2.97 mL, 11.86 mmol, 8.43 equiv) was added. The reaction was complete after 60 minutes (HPLC). The reaction was completed in roto-evaporator at 0 ° C. To this was added IPA (2 mL) then hydroxylamine solution (1859 mL, 28.1 mmol, 20 equiv). It was placed in a flask at 4 ° C for ~ 40h. The reaction was concentrated, (keeping the reaction 0 ° C) to gummy mass. To this was added water (3 mL) and ACN (0.5 mL). It was acidified while it is at 0 ° C using TFA (3 mL). Additional water (1 mL) and ACN (1 mL) were added. It was purified by RP HPLC (2"column, 50mL / min, 0.1% TFA in gua / ACN, equil to 5% B), and loaded on a 2" column (10mL / min, 5% B) using a syringe (2X6.5 mL) was increased to 50 mL / min for 1 minute, 5-30% B for 66 minutes, and eluted to 25% B. The desired fractions were combined, frozen and placed in ice. 410 mg of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2R) -2- (hydroxymethyl) -2 -methylcyclopropyl) buta-1,3-diinyl) benzamide (1-11), TFA LCMS M + l expected = 384.2, observed = 384.2 NMR-1H (D SO-d6): d 0.61-0.64 (t, 1H ), 1.02-1.06 (m, 1H), 1.16 (s, 3H), 1.25 (s, 3H), 1.30 (s, 3H), 1.55-1.60 (m, 1H), 3.17-3.29 (m, 2H), 4.65-4.67 (d, 1H), 7.61-7.63 (d, 2H), 7.88-7.90 (d, 2H), 7.98 (s, 2H), 8.53-8.56 (d, 1H), 9.21 (br, 1H), 11.19 (s, 1H).

J. Synthesis of N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-cis) -3- (hydroxymethyl) ) cyclobutyl) buta-1,3-diinyl) benzamide (1-12) N- ((S) -3-amino-1- (idroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ( ((1, 3-trans) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) benzamide (1-13) 3-Methylenecyclobutanecarboxylic acid (2) The solution of 3-methylenecyclobutanecarbonitrile 1 (27.9g, 0.3mol) in ethanol (200mL) and water (200mL) was treated with potassium hydroxide (84g, 1.5mol). The resulting mixture was stirred and heated at 105 ° C for 4 h. The ethanol was removed under reduced pressure. The residue was cooled to 0 ° C, and the pH was adjusted to 1-2, then extracted with ethyl acetate, dried over sodium sulfate4 / filtered and the solvent was removed. (30.2g, yield = 90%). 3 - . 3 - . 3-methyl-methylenecyclobutanecarboxylate (3) The mixture of 3-methylenecyclobutanecarboxylic acid 2 (29.12g, 0.26mol), potassium carbonate (70.2g, 0.52mol), acetone (500mL) and dimethyl sulfate (39.312g, 0.312mol) was heated to reflux for 2 h. The reaction mixture was cooled to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified with silica gel chromatography to give the desired product as a colorless oil. (27.5g, yield = 84%). 3 - . 3 - Methyl (hydroxymethyl) cyclobutanecarboxylate (4) Three-necked, dry flask was charged with methyl methylenecyclobutanecarboxylate 3 (24.46g, 0.21mol) and dry THF (130ml) and cooled to -10 ° C, then borane-THF complex (70.OmL) was added through a drop by drop syringe. The resulting mixture was stirred for 4 hours at room temperature and cooled to -20 ° C ~ -10 ° C; methanol was added and stirred for 15 minutes. Sodium hydroxide (3M 30mL) and hydrogen peroxide (34.Og, 0.210mol) were added in sequence. The mixture was stirred for 2 hours and saturated sodium sulfite solution (100mL) was added. The reaction mixture was diluted with water, then extracted with ethyl acetate, washed with water and brine, dried over sodium sulfate, filtered, the solvent was removed and the residue was purified by silica gel chromatography. (28.73g, 95%) 3-formylcyclobutanecarboxylate (5a) and (cis) -methyl 3- formylcyclobutanecarboxylate (trans) -methyl (5b) To a solution of oxalyl chloride (17.8g, 139mmol, 2. Oeq) in CH2C12 (450mL) was slowly added DMSO (21.7g, 278mmol, 4. Oeq) in dichloromethane (50mL) at -78 ° C. After 30 minutes, methyl 3- (hydroxymethyl) cyclobutanecarboxylate 4 (10g, 69.4mmol, l.Oeq) in dichloromethane (150mL) was added dropwise at -78 ° C. The mixture was stirred for an additional 2 h at -78 ° C, and then at -78 ° C triethylamine (70 g, 700 mmol, 10. Oeq) was added. After 20 min, the mixture was warmed to room temperature, and saturated aqueous NH 4 Cl was added. The layers were separated and the aqueous layer was extracted with dichloromethane (3 X 200mL). The combined organic layers were washed with brine, dried (sodium sulfate), filtered and concentrated to give the crude product as a mixture of cis and trans isomers. Purification to obtain 5a (trans, 3.1 g) and 5b (cis, 2.9 g) per silica column. 3- (2, 2-dibromovinyl) cyclobutanecarboxylate (cis) -methyl To a solution of carbon tetrabromide (13.56g, 40.89mmol, 2. Oeq) in 40mL of dichloromethane, a solution of triphenylphosphine (21.41g, 81.80mmol, 4. Oeq) in 80mL of dichloromethane was added dropwise in a dropwise manner. -20 ° C under argon. The mixture was kept stirring at -20 ° C for 30 min, then cooled to -78 ° C. 3-Formylcyclobutanecarboxylate (cis) -methyl 5b (2.9g, 20.40mmol, l.Oeq) in 80mL of dichloromethane was added dropwise and kept stirring at -78 ° C for another hour. The mixture was allowed to warm to room temperature. The reaction mixture was added dropwise with stirring at 300 mL of PE. The mixture was filtered and the residue was washed with PE. The combined organic layer was dried over sodium sulfate and concentrated to give the objective compound which was purified by column of silica gel. (2.3g, yield = 37%) ((cis) -3- (2,2-dibromovinyl) cyclobutyl) methanol (7b) Compound 6b (2.5g, 8.4mmol, l.Oeq) dissolved in THF (10 mL) was added lithium borohydride (203 mg) at 0 ° C. The reaction mixture was kept stirring for 3 h at room temperature, water (5mL) was added and it was extracted with ethyl acetate. The combined ethyl acetate layer was dried and concentrated under reduced pressure. The crude product was purified by column on silica gel with PE: EA = 5: 1. (1.88g yield = 83%). 4- (((cis) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) methyl benzoate (9b) A mixture of 1 ((cis) -3- (2, 2-dibromovinyl) -cyclobutyl) methanol 7b (1.88g, 6.9mmol, l.Oeq), methyl 4-ethynylbenzoate 8 (1.56g, 9.7mmol, 1.4eq) ), tri (4-methylphenyl) phosphine (98mg, 0.28mmol, 0.04eq), Pd (dba) 4 (64mg, 0. 069mmol, O.Oleq) in dry DMF (30mL) was added triethylamine (2.11g, 20.9mmol, 3. Oeq) under nitrogen atmosphere. The reaction mixture was kept stirring at 80 ° C for 15 h. The LCMS monitored the reaction. The reaction mixture was diluted with NH4Clac solution and extracted with ethyl acetate. The combined ethyl acetate layer was washed with water, brine and dried. The solvent was evaporated under reduced pressure to give to give the crude product which was purified by column on silica gel with PE: EA = 5: 1. (450mg, yield = 24%). 4- (((cis) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) -benzoic acid (ACHL02-146-cis) To a solution of methyl 4- (((cis) -3- (hydroxymethyl) -cyclobutyl) buta-1,3-diinyl) benzoate 9b (450 mg, 1.0 equivalent) in THF (200 mL) was added monohydrate hydroxide of lithium (282mg, 4.0 equivalents), then the mixture was reacted at 25 ° C for 5h. Then the reaction solvent was removed and neutralized with 3 N HC1 to pH 5-6, extracted with ethyl acetate. The combined ethyl acetate layer was washed with brine and dried. The solvent was evaporated to obtain the parent compound. (400mg, yield = 94%). 3- (2, 2-Dibromovinyl) cyclobutanecarboxylate of (trans) -methyl (6a) To a solution of carbon tetrabromide (11.2g, 33. 80mmol, 2. Oeq) in 40mL of dichloromethane, a solution of triphenylphosphine (17.7g, 67.60mmol, 4. Oeq) in 80mL of dichloromethane was added dropwise below -20 ° C under argon. The mixture was kept stirring at -20 ° C for 30 min, then cooled to -78 ° C. 3-Formylcyclobutane carboxylate (trans) -methyl 5a (2.4gf 16.90mmol, 1. Oeq) was added dropwise in 80mL of dichloromethane, and was kept stirring at -78 ° C for another lh. The mixture was allowed to warm to room temperature the reaction mixture was added dropwise to a PE of 300mL with stirring. It was filtered and the residue was washed with PE. The combined organic layer was dried over sodium sulfate and concentrated to give the parent compound which was purified by column on silica gel. (2.5g, yield = 50%). ((trans) -3- (2,2-Dibromovinyl) cxclobutyl) methanol (7a) Reagent PM Eq. Mmol g, mL Compound 6a 298 1.0 8.4 2.5g LÍBH4 22 1.1 9.2 203mg THF lOmL 3 - (2,2-dibromovinyl) cid .obutane carboxylate of (trans) -methyl 6a (2.5g, 8.4mmol, l.Oeq) was dissolved in THF (10mL), lithium borohydride (203mg) was added at 0 °. C. The reaction mixture was kept stirring for 3 h at room temperature, water (5 mL) was added and it was extracted with ethyl acetate. The combined ethyl acetate layer was dried and concentrated under reduced pressure. The crude product was purified by column on silica gel with PE: EA = 5: 1. (1.70g, yield = 75%). 4- (((trans) -3- (Hydroxymethyl) cyclobutyl) buta-1,3-diinyl) -benzoic acid methyl ester (9a) A mixture of ((trans) -3- (2,2-dibromovinyl) -cyclobutyl) methanol 7a (1.70g, 6.3mmol, l.Oeq), compound 8 (1.41g, 8.8mmol, 1.4eq), tri ( 4-methylphenyl) phosphine (89mg, 0.25mmol, 0.04eq), Pd (dba) 4 (58rag, 0.063mmol, O.Oleq) in dry DMF (30mL) was added triethylamine (1.91g, 18.9mmol, 3.0eq) under nitrogen atmosphere. The reaction mixture was kept stirring at 80 ° C for 15 h. LC S monitored the reaction. The reaction mixture was diluted with NH4Clac solution and extracted with ethyl acetate. The combined ethyl acetate layer was washed with water, brine and dried. The solvent was evaporated under reduced pressure to give the crude product which was purified by column on silica gel with PE: EA = 5: 1. (550mg, yield = 33%). 4- (((trans) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) benzoic acid (ACHL02-146-trans) To a solution of methyl 4- (((trans) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) benzoate 9a (550 mg, 1.0 equivalent) in THF (20 mL) was added monohydrate hydroxide lithium (345mg, 4.0 equivalents), then the mixture was reacted at 25 ° C for 5h. Then the reaction solvents were removed and neutralized with 3 N HCl to pH 5-6, extracted with ethyl acetate. The combined ethyl acetate layer was washed with brine and dried. The solvent was evaporated to obtain the parent compound. (340mg, yield = 65%). 2-Amino-3- (tert-butoxycarbonyl-amino) -3-methylbutanoate of (S) -methyl (387 mg, 1573 mmol, 1.0 equivalents) and 4- (((1,3-cis) -3) were dissolved. - (hydroxymethyl) -cyclobutyl) buta-1,3-diinyl) benzoic acid (400 mg, 1573 mmol, 1.0 equivalent) in DMF (3 mL). To this was added DIPEA (0.687 mL, 3.93 mmol, 2.5 equivalents) then HATU (718 mg, 1.888 mmol, 1.2 equivalents). This was stirred at room temperature for -16 h. The reaction was partitioned between 1M citric acid and ethyl acetate. The organic products were washed with semi-saturated sodium chloride, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. It produced 1,035 g of 3- (tert-butoxycarbonylamino) -2- (4- (((1,3-cis) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate from ( S) -methyl crude. LCMS M + l expected = 483.2, observed = 483.2. 3- (tert-Butoxycarbonylamino) -2- (4- (((1,3-cis) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (S) -methyl (crude) was dissolved in methanol (1 mL). At room temperature, HC1 4. OM in dioxane (3.3 mL, 13.26 mmol, 8.43 equivalents) was added. The reaction is complete after 60 minutes (HPLC). The reaction was concentrated on a rotoevaporator at 0 ° C. To this was added IPA (2 mL) then hydroxylamine solution (2 mL, 31.5 mmol, 20 equivalents). The flask was placed at 4 ° C for 24 h. The reaction was concentrated, (keeping the reaction at 0 ° C) to gummy mass. To this was added water (3 mL) and ACN (0.5 mL). Acidify as it is at 0 ° C using TFA (3 mL). Additional water (1 mL) and ACN (1 mL) were added. HPLC purity of RP (column 2", 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 5% B) was loaded on a 2" column (lOmL / min, 5% B) using syringe filter (2X6 mL). It was increased to 50mL / min for 1 minute. 5-35% B for 55min, product eluted at 24.5-26% B. The desired fractions were combined, frozen and placed in lyophilisate. Produced 460 mg of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-cis) -3- (hydroxymethyl) Cyclobutyl) buta-1,3-diinyl) benzamide, TFA. LCMS M + l expected = 384.2, observed = 384.2. RM ^ H (DMSO-d6): d 1.25 (s, 3H), 1.30 (s, 3H), 1.81-1.88 (m, 2H), 2.23-2.34 (m, 3H), 3.08-3.15 (m, 1H) , 3.30-3.31 (d, 1H), 4.54 (br, 1H), 4.65-4.67 (d, 1H), 7.61-7.64 (d, 2H), 7.89-7.91 (d, 2H), 7.99 (s, 2H) , 8.54-8.56 (d, 1H), 9.22 (br, 1H), 11.20 (s, 1H). dissolved 2-amino-3- (tert-butoxycarbonyl amino) -3-methylbutanoate of (S) -methyl (329 mg, 1377 mmol, 1.0 equivalent) and 4- (((1, 3-trans) -3- ( hydroxymethyl) -cyclobutyl) buta-1,3-diinyl) benzoic acid (340 mg, 1377 mmol, 1.0 equivalent) in DMF (3 mL). To this was added DIPEA (0.584 mL, 3.34 mmol, 2.5 equivalents) then HATU (610 mg, 1.605 mmol, 1.2 equivalents). This was stirred at room temperature for -16 h. The reaction was partitioned between 1M citric acid and ethyl acetate. The organic products were washed with semi-saturated sodium chloride, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. Produced 990 mg of 3- (tert-butoxycarbonylamino) -2- (4- (((1,3-trans) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) venza-mido) -3-methylbutanoate of (S) -methyl crude. LCMS M + l expected = 483.2, observed = 483.2. 3- (tert-butoxycarbonylamino) -2- (4- (((1,3-trans) -3- (hydroxymethyl) cyclobutyl) buta-1,3-diinyl) -benzamido) -3-methylbutanoate was dissolved (S) ) -methyl (crude) in methanol (1 mL). HCl 4. OM in dioxane (2.82 mL, 11.27 mmol, 8.43 equivalents) was added at room temperature. The reaction was complete after 60 minutes (HPLC). The reaction was concentrated on a rotoevaporator at 0 ° C. To this was added IPA (2 mL) then hydroxylamine solution (1.77 mL, 26.7 mmol, 20 equivalents). The flask was placed at 4 ° C for 96h. Then the reaction was concentrated (keeping the reaction at 0 ° C) to a gummy mass. To this was added water (3 mL) and ACN (0.5 mL). The solution was acidified at 0 ° C using TFA (3 mL). Additional water (1 mL) and ACN (1 mL) were added. The compound was purified by RP HPLC (column 2", 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 5% B); loaded on a 2"column (lOmL / min, 5% B) using a syringe filter (2X7 mL), increased to 50mL / min for 1 minute and 5-35% B for 55min.The product was eluted at 22.9-25.5 % B The desired fractions were combined, frozen and placed in lyophilisate.The procedure yielded 410 mg of N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-1-oxobutan-2. il) -4- (((1,3-trans) -3- (hydroxymethyl) -cyclobutyl) uta-1,3-diinyl) benzamide, TFA, LCMS M + l expected = 384.2, observed = 384.2, NMRH (DMSO) -d6): d 1.25 (s, 3H), 1.30 (s, 3H), 2.07-2.11 (t, 4H), 2.40-2.43 (m, 1H), 3.22-3.26 (m, 1H), 3.37-3.38 ( d, 1H), 4.57 (br, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H), 7.89-7.91 (d, 2H), 7.97 (s, 2H), 8.54-8.56 ( d, 1H), 9.21 (br, 1H), 11.19 (s, 1H).

K Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-trans) -3- (hydroxymethyl) cyclopentyl) ) -buta-1,3-diinyl) benzamide (1-14) ACHL02-149 (Cis) -cyclopentane-1,3-dicarboxylic acid (2) To a stirred suspension of potassium permanganate (47 g, 0.297 mol) and magnesium sulfate (4.3 g, 35.4 mmol) in water (500 mL) cooled to 5 ° C was added a solution of bicyclo [2.2.1] hepty- 2-ene 1 (10 g, 0.106 mol) in acetone (15 mL) in one portion (5 ° C -40 ° C). The ice bath was stirred and the reaction was stirred for an additional 2 h. After this time, the mixture was filtered and the filtrate was treated with sodium hydrogensulfite and acidified to pH = 2 with 1N HC1. It was then extracted with ethyl acetate (800 mL x 3) and the combined organic layer was washed with water (1 L), brine (1 L), dried, concentrated to give the starting compound 2 (7.5 g, 46 g. %) as a white solid without purification for the next step.

Cyclopentane-1,3-dicarboxylic acid (cis) -dimethyl ester (3) a stirred solution of (cis) -cyclopentane acid 1,3-dicarboxylic acid 2 (7.5 g, 47.5 mmol) in methanol (200 mL) was added thionyl chloride (22.6 g, 0.19 mol) in an ice bath. Then the reaction mixture was heated to reflux overnight. After this time, the mixture was concentrated to give compound 3 (8.3 g, 94%) as a yellow oil without purification for the next step. Confirmed by GC-MS.

Cyclopentane-1,3-diyldimethanol (4) To a stirred solution of LAH (3.4 g, 89.2 mmol) in THF (100 mL) was added a solution of cyclopentane-1,3-dicarboxylic acid (cis) -dimethyl 3 (8.3 g, 44.6 mmol) in THF (50 mL ) in an ice bath. Then the reaction mixture was stirred at room temperature overnight. After this time, the mixture was diluted with water (6.5 mL), then filtered and the filtrate was concentrated, the residue was purified by silica gel chromatography (PE: EA = 2: 1) to give compound 4 ( 4.0 g, 69%) as a yellow oil. NMR (400 MHz, DMSO-d6) 0.53-0.50 (m, 1H), 0.96-1.00 (m, 2H), 1.29-1.35 (m, 2H), 1.47-1.54 (m, 1H), 1.66-1.73 (m , 2H), 2.98-3.01 (m, 4H), 4.29 (s, 1H). 3- (Hydroxymethyl) cyclopentyl) methyl acetate (5) a stirred solution of cyclopentane-1,3-diylmethanol-4 (4.0 g, 31 mmol) in THF (60 mL) was added NaH (60%) (1.1 g, 28 mol) in an ice bath. Then the reaction mixture was stirred in an ice bath for 0.5 h, then acetyl chloride (2.7 g, 34 mmol) was added to the reaction mixture. And the mixture was stirred at room temperature overnight. After this time, the mixture was diluted with water (1.0 mL), then filtered, and the filtrate was concentrated, the residue was purified by silica gel chromatography to give compound 5 (1.7 g, 25%) as a yellow oil NMR (400 MHz, DMSO-d6) 0.84-0.87 (m, 1H), 1.28-1.31 (m, 2H), 1.62-1.65 (m, 2H), 1.80-1.83 (m, 1H), 1.99 (S, 1H) ), 2.10-2.18 (m, 1H), 3.26-3.29 (m, 2H), 3.89 (d, J = 8, 2H), 4.43 (s, 1H). 3-formylcyclopentyl) methyl acetate (6) a stirred acetate solution (hydroxymethyl) cyclopentyl) methyl 5 (1.7 g, 9.9 mmol) in CH2C12 (50 mL) was added PCC (6.4 g, 29.7 mmol) in portions. Then the reaction mixture was stirred at room temperature overnight. After this time, the mixture was concentrated, the residue was purified by silica gel chromatography (PE: EA = 6: 1) to give compound 6 (0.7 g, 44%) as a yellow oil. 3-Ethynylcyclopentyl) methanol (7) To a stirred solution of 3-formyl-cyclopentyl acetate) methyl 6 (0.7 g, 4.1 mmol) in MeOH (50 mL) was added potassium carbonate (0.85 g, 6.15 mmol) and Bestmann's Reagent (1.18 g, 6.15 g). mmol). The reaction mixture was stirred at room temperature overnight. After this time, the mixture was diluted with water (50 mL), extracted with ether (80 mL x 3) and the combined organic layer was dried, concentrated, the residue was purified with silica gel chromatography (PE-.EA = 2: 1) to give compound 7 (0.31 g, 60%) as a yellow oil.

Methyl 4- ((3- (hydroxymethyl) cyclopentyl) buta-1,3-diinyl) benzoate (ACHL02-149) Under nitrogen, DIPEA (936 mg, 7.26 mmol) was added dropwise to a mixture of 3-ethynylcyclopentyl) -methanol 7 (0.3 g, 2.42 mmol), methyl 4- (bromoetinyl) benzoate 8 (0.7 g, 2.9 mmol ), PdCl2 (PPh3) 2 (70mg, 0.1 mmol) and Cul (46mg, 0.242 mmol) in THF (60 mL) at RT. The reaction mixture was stirred at RT overnight. The LCMS monitored the reaction. The reaction mixture was diluted with water (60 mL) and extracted with ethyl acetate (80mL x 3). The ethyl acetate layer was washed with saturated aqueous NH 4 Cl solution, water and brine. After drying, the ethyl acetate solution was concentrated under reduced pressure. The residue was purified with silica gel column (PE: EA = 5: 1) to obtain compound ACHL02-149 (as a mixture of cis and trans isomers) (140 mg, 25%). H NMR (400MHz, DMSO-d6): 7.95 (d, J = 8.4Hz, 2H), 7.51 (d, J = 8Hz, 2H), 3.91 (s, 1H), 3.61 (d, J = 4Hz, lH) , 3.53 (d, J = 8Hz, 1H), 2.89-2.87 (m, 1H), 2.43-2.40 (m, 1H), 2.21-2.17 (m, 1H), 2.02-1.99 (m, 1H), 1.82- 1.76 (m, 1H), 1.54-1.45 (m, 1H), 1.36-1.31 (m, lH).

Methyl 4- ((3- (hydroxymethyl) cyclopentyl) buta-1,3-diinyl) benzoate (115 mg, 0.407 mmol, 1.0 equivalent) was dissolved in MeOH (5 mL), followed by the addition of lithium hydroxide (29.3 mg, 1222 mmol, 3.0 equivalents) and water (5 mL). This was stirred at room temperature for ~72 h. The reaction was concentrated to remove the MeOH, acidify to pH ~ 3 with HC1 6N (~ 5mL). Extract with EtOAc (3x50 mL), combine the organic layers, wash with saturated NaCl, dry (MgSO4), filter, concentrate. It produced 0.14 g of crude 4- ((3- (hydroxymethyl) cyclopentyl) buta-1,3-diinyl) benzoic acid. LCMS M + l sperado = 269.1, observed 2-Amino-3- (tert-butoxycarbonyl-amino) -3-methylbutanoate of (S) -methyl (100 mg, 0.406 mmol, 1.0 equivalent) and 4- ((3- (hydroxymethyl) cyclopentyl) -buta acid were dissolved. -1, 3-diinyl) benzoic acid (crude) (theoretical 109 mg, 0.406 mmol, 1.0 equivalent) in DMF (l mL). To this was added DIPEA (0.177 mL, 1.016 mmol, 2.5 equivalents) then HATU (185 mg, 0. 488 mmol, 1.2 equivalents). This was stirred at room temperature for -16 h. The reaction was partitioned between 1M citric acid and ethyl acetate. The organic products were washed with semi-saturated sodium chloride, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. It produced 500 mg of crude (S) -methyl 3- (tert-butoxycarbonylamino) -2- (4- ((3- (hydroxymethyl) cyclopentyl) -buta-1,3-diinyl) benzamido) -3-methylbutanoate. LCMS M + l expected = 497.3, observed = 497.2. 3- (tert-Butoxycarbonylamino) -2- (4- ((3- (hydroxy-methyl) cyclopentyl) uta-1,3-diinyl) benzamido) -3-methylbutanedioate of (2S) -methyl (crude) is dissolved in methanol (1 mL). At room temperature 4.0M HCl in dioxane (0.857 mL, 3.43 mmol, 8.43 equivalents) was added. The reaction is complete after 3 hours (HPLC). The reaction was concentrated on a rotoevaporator at 0 ° C. To this was added IPA (1 mL) then hydroxylamine solution (0.537 mL, 8.14 mmol, 20 equivalents). Place the flask at 4 ° C for 168h. The reaction was concentrated, (keeping the reaction at 0 ° C) to gummy mass. To this was added water (3 mL) and ACN (0.5 mL). Acidified while it is at 0 ° C using TFA (3 mL). Additional water (1 mL) and ACN (1 mL) were added. It was purified by RP HPLC (1"column, 20mL / min, 0.1% TFA in water / ACN, equilibrated @ 5% B), loaded on a 1" column (lOmL / min, 5% B) using a filter of syringe (2X8 mL). It was increased to 20mL / min for 1 minute. 5-19% B for 80min, the product elutes at 19% B. The desired fractions were combined, frozen and placed in lyophilisate. Produced 145 mg of N - ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ((3- (hydroxy-methyl) cyclopentyl) buta-1 , 3-diinyl) benzamide, 1-14, TFA. LCMS M + l expected = 398.2, observed = 398.2. H-NMR (DMSO-d6): d 1.25 (s, 3H), 1.29 (s, 3H), 1.38-1.45 (m, 1H), 1.53-1.77 (m, 2H), 1.88-2.17 (m, 3H), 2.83-2.91 (m, 1H), 3.21-3.38 (m, 4H), 4.52 (br, 1H), 4.65-4.67 (d, 1H), 7.61-7.63 (d, 2H), 7.88-7.90 (d, 2H) ), 7.97 (s, 2H), 8.53-8.56 (d, 1H), 9.21 (br, 1H), 11.19 (s, 1H).

L. Synthesis of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,4-cis) -4- (hydroxymethyl) ) cyclohexyl) -buta-1,3-diinyl) benzamide (1-15) and N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 - (((1, 4-trans) -4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) benzamide (1-16) Step 1: Dimethyl cyclohexane-1,4-dicarboxylate (2) Methanol (500 mL) was cooled in an ice salt bath and thionyl chloride (138 g, 4.0eq) was added dropwise. To the resulting solution of HC1 in methanol, cyclohexane-1,4-dicarboxylic acid 1 (50g, 1.0 equivalent) was added and the reaction mixture was heated to reflux for 1 h. TLC monitored the reaction. The solvent was evaporated and the residue was diluted with water, extracted with ethyl acetate. The ethyl acetate layer was washed with NaHCO3 solution > brine and dried over Na2SO4. The solvent was evaporated to obtain the parent compound (56.4g, 97%).

Step 2: Cyclohexane-1,4-diyldimethanol (3) An ice-cooled suspension of LiAlH4 in THF was added dropwise a solution of dimethyl cyclohexane-1,4-dicarboxylate 2 at 0 ° C. The reaction mixture was kept stirring at ambient temperature for 30 min. TLC monitored the reaction. Water 12eq (32.4mL) was added to quench the reaction. The reaction mixture was filtered, and the filtrate was concentrated. The residue was diluted with Et20, dried over Na2SO4. Et20 was removed to collect the parent compound that was used for the next step without further purification (19.9g, 92%).

Step 3: (4- (Hydroxymethyl) cyclohexyl) methyl acetate An ice-cooled suspension of NaH and cyclohexane-1,4-diylmethanol-3 in THF was added dropwise AcCl at 0 ° C. The reaction was kept stirring for 3 h at 15 ° C. TLC monitored the reaction. H20 1 eq (2.5mL) was added to quench the reaction. The reaction mixture was filtered, and the filtrate was diluted with ethyl acetate. The ethyl acetate layer was washed with brine (5mL), dried over Na2SO4. The solvent was removed under reduced pressure. The residue was purified with silica gel column (DCM: MeOH = 50: 1 to 10: 1) to obtain the parent compound (8.6g, 33%).

Step 4: (-formylcyclohexyl) methyl acetate (5) To a solution of oxalyl chloride in DCM (60 mL) was added dropwise DMSO in DCM (60 mL) at -78 ° C. After 30 min, (4- (hydroxymethyl) -cyclohexyl) methyl 4 acetate in DCM (80mL) was added dropwise at -78 ° C. The reaction mixture was stirred for an additional 2 h at -78 ° C, then TEA was added dropwise at -78 ° C to quench the reaction. The reaction mixture was allowed to warm to room temperature. The reaction mixture was diluted with DCM, and washed with saturated aqueous ammonium chloride and brine. The DCM layer was dried and concentrated. The residue was purified with a silica gel column (DCM) to obtain the original compound (9.0 g, still wet with dichloromethane).

Step 5: (4-Ethynylcyclohexyl) methanol (6) methyl 5 (8.5g, l.Oeq), K2C03 (19.04g, 3. Oeq) in CH3OH (100mL) was added Bestmann's reagent (11.56g, 1.3eq, 80%) at 0 ° C. The mixture was stirred at room temperature for 15 h. TLC monitored the reaction. The mixture was diluted with water, extracted with ethyl acetate, washed with brine, dried over Na 2 SO 4, concentrated under reduced pressure. The crude product was purified with silica gel column (PE: EA = 3: 1) to obtain the parent compound (2.6g, 40%).

Step 6: 4- (Methyl 4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) -benzoate (8) Under nitrogen, DIPEA was added dropwise (3.92g, 3.0 equivalents) to a mixture of (4-ethynylcyclohexyl) methanol 6 (1.4g, 1.0 equivalents), methyl 4- (bromoethinyl) -benzoate 7 (2.9g, 1.2 equivalents), PdCl2 (PPh3) 2 (213mg, 0.03 equivalents) and Cul (194mg, 0.1 equivalents) in THF (150mL) at RT. The reaction mixture was stirred at RT overnight. The LCMS monitored the reaction. The reaction mixture was diluted with water (100mL) and extracted with ethyl acetate (300mL). The ethyl acetate layer was washed with saturated aqueous NHC1 solution, water and brine. After it was dried, the ethyl acetate solution was concentrated under reduced pressure. The residue was purified with silica gel column (PE: EA = 5: 1 to 3: 1) to obtain the parent compound (as a mixture of cis and trans isomers, 1.2 g, 40%). H-NMR (400MHz, DMSO-d6): 7,959-7,937 (d, J = 8.8Hz, 2H), 7,682-7,660 (d, J = 8.8Hz, 2H), 4. 421-4,395 (t, J = 5.2Hz, 1H), 3860 (s.3H), 3.207-3.179 (t, J = 5.6Hz, 2H), 2085 (s, 1H), 1980-1941 (m, 2H) , 1. 747-1.708 (m, 2H), 1366-1.328 (m, 3H), 0.935-0.897 (m, 2H). 4- ((4- (hydroxymethyl) cyclohexyl) buta-l, 3-diinyl) - To a solution of methyl 4- ((4- (hydroxymethyl) cyclohexyl) buta-1, 3-diinyl) benzoate 1 (1.2g, 4.1mmol) in THF (20mL) was added a solution of monohydrate lithium hydroxide (517mg, 12.3mmol) in water (5mL). The mixture was kept stirring at room temperature overnight. After this time, the solvent was removed under reduced pressure, the residue was diluted with water (40mL), the pH adjusted to 4.0 with 1M HCl, then extracted with ethyl acetate (3 x 60mL). The organic layer was dried and concentrated to give 4- ((4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) enzoic acid 2 (1.0 g, 93%) as a yellow solid which was used for the next step without additional purification. LC-MS: 283 [M + H] +. 3- (tert-Butoxycarbonylamino) -2- (4- ((4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (S) -methyl (3) a solution of 4- ((4- (hydroxymethyl) cyclohexyl) buta-l, 3-diinyl) benzoic acid 2 (1.05g, 3.72mmol), HATU (1.55g, 4.1mmol) in DMF (25mL) was added 2- amino-3- (tert-butoxycarbonylamino) -3-methylbutanoate of (S) -methyl (915 mg, 3.72 mmol) and DIPEA (1.44 g, 11.2 mmol). The mixture was kept stirring at room temperature for 5 hours. After this time, the reaction was diluted with water (20mL), then extracted with ethyl acetate (60mL x 3). The combined organic layers were washed with water then brine, dried and concentrated, the residue was purified with silica gel chromatography (PE: EA 2: 1) to give 3- (tert-butoxycarbonylamino) -2- (4- ((4- (hydroxymethyl) -cyclohexyl) buta-1,3-diinyl) -enoxamido) -3-methylbutanoate (S) -methyl 3 as a yellow solid. (1.6 g, 85%). LC-MS: 511 [M + H] + .455 [M-56] 3-amino-2- (4- ((4- (hydroxymethyl) cyclohexyl) -buta-1,3-diinyl) -benzamido hydrochloride) -3-methylbutanoate of (S) -methyl (4) A solution of (S) -methyl-3- ((4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate of (S) -methyl 3 (1.6g, 3.14mmol) in methanol (20mL) was attached to an HC1 apparatus. Then the reaction was stirred at room temperature until TLC indicated the total consumption of the starting material. After that, the solution was concentrated under reduced pressure to give 3-amino-2- (4- ((4- (hydroxymethyl) cyclohexyl) -buta-1,3-diinyl) -benzamido) -3-methylbutanoate hydrochloride. (S) -methyl 4 as a yellow solid (1.4 g, 99%). LC-MS: 411 [M + H] +. The cis and trans isomers were separated by chiral HPLC to give 3-amino-2- (4- (((1,4-cis) -4- (hydroxymethyl) cyclohexyl) uta-1,3-diinyl) benzamido) -3 -methylbutanoate (S) -methyl (ACHL02-148-coupled, 90 mg) as peak 1 and 3-amino-2- (4- (((1,4-trans) -4- (hydroxymethyl) cyclohexyl) buta- 1, 3-diinyl) -benzamido) -3-methylbutanoate (S) -methyl (ACHL02-147-coupled, 750 mg) as peak 2. 3-Amino-2- (4- (((1, -trans) -4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) benzatnido) -3-methyl-butanoate of (S) -methyl (750 mg , 1.827 mmol, 1.0 equivalents) was added to a round bottom flask. To this was added IPA (6 mL), the suspension was sonicated for 5 minutes, then THF (2 mL) was added to form a solution. Hydroxylamine solution (2.4 mL, 36.5 mmol, 20 equivalents) was added and the flask was placed at 4 ° C for ~ 96h. The reaction was concentrated, (keeping the reaction at 0 ° C) to remove IPA and THF. Acidified as it is at 0 ° C using TFA (4 mL). Additional ACN (2mL) was added. It was purified by RP HPLC (2"column, 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 10% B) .It was loaded on a 2" column (lOmL / min, 10% B) using a filter. of syringe (3X8 mL). It was increased to 50mL / min for 1 minute. 10-40% B for 60min, product eluted at 30.5-35% B. The desired fractions were combined, frozen and placed in lyophilisate. Produced 706 mg of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1, A-trans) -4- (hydroxymethyl) ) cyclohexyl) buta-1,3-diinyl) benzamide, 1-16, TFA. LCMS M + l expected = 412.2, observed = 412.2. XH NMR (DMSO-d6): d 0.84-0.94 (m, 2H), 1.25 (S, 3H), 1.30 (s, 3H), 1.32-1.36 (m, 1H), 1.68-1.72 (m, 2H), 1.90-1.95 (m, 2?), 2.39-2.42 (m, 1?), 3.15-3.18 (m, 2H), 4. 38-4.40 (t, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H), 7. 89-7.91 (d, 2H), 7.98 (br (2H), 8.54-8.56 (d, 1H), 9.22 (br, 1H), 11.21 (br, 1H). 3-Amino-2- (4- (((1, -cis) -4- (hydroxymethyl) cyclohexyl) buta-1,3-diinyl) benzamido) -3-methylbutanoate (Sl-methyl (90 mg, 0.219 mmol , 1.0 equivalents) was added to a round bottom flask, IPA (0.5 mL) was added, the solution was formed in a vortex for 2 minutes, then hydroxylamine solution (0.29 mL, 4.38 mmol, 20 equivalents) was added and The flask was placed at 4 ° C for ~ 72 h.The reaction was concentrated, (keeping the reaction at 0 ° C) to remove IPA, acidified to 0 ° C using TFA (3 mL), acetonitrile was added. (0.5 mL) was purified by RP HPLC (1"column, 25mL / min, 0.1% TFA in water / ACN, equilibrated @ 10% B), loaded on a 1" column (10mL / min, 10). % B) using syringe filter (7 mL), was increased to 25mL / min for 1 minute, 10-40% B for 90min, elucidated to 25.5-28% B. The desired fractions were combined, frozen and placed in lyophilized, produced 53 mg of N- ((S) -3-amino-l- (hydroxy amino) -3-methyl-l-oxobutan-2-yl) -4- (((1,4-cis) -4- (hydroxymethyl) cyclohexyl) buta-1, 3- diinyl) benzamide, 1-15, TFA. LCMS M + l expected = 412.2, Observed = 412.2. XH NMR (DMSO-d6): d 1.20-1.35 (m, 2H), 1.26 (s, 3H), 1.30 (s, 3H), 1.47-1.59 (m, 4H), 1.71-1.75 (m, 2H), 2.97-2.99 (m, 1H), 3.21-3.22 (d, 2H), 4.30 (br, 1H), 4.65-4.68 (d, 1H), 7.64-7.66 (d, 2H), 7.89-7.91 (d, 2H) ), 7.98 (br, 2H), 8.54-8.56 (d, 1H), 9.22 (br, 1H), 11.20 (s, 1H).

M. Synthesis of N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3 trifluoroacetate -diinyl) benzamide (1-17) and N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((cis) -2- (hydroxymethyl) trifluoroacetate ) cyclopropyl) -buta-1, -diinyl) benzamide (1-18) 4- (((trans) -2- (hydroxymethyl) cyclopropyl) -buta-l, 3-diinyl) benzoic acid (205 mg, 0.85 mmol, 1 eq) and (S) -methyl-2-amino-2- ( 1- (tert-butoxycarbonylamino) cyclobutyl) -acetate (220 mg, 0.85 mmol, 1 eq) were dissolved in N, N-dimethylformamide (3mL). N, iV-diisopropylethylamine (0.45mL, 2.6mmol, 3eq) was added, followed by HATU (356mg, 0.94mmol, 1. leq). The mixture was kept at room temperature for 2.5 hours, then it was partitioned between 1M citric acid and ethyl acetate. The organic products were washed with 1M citric acid, saturated sodium bicarbonate then chloride Saturated sodium, dried over magnesium sulfate and evaporated to dryness to give 2- (1- (tert-butoxycarbonyl-amino) cyclobutyl) -2- (4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta -1,3-diinyl) benzamido) (S) -methyl acetate (crude 550mg, -134%, was used without further purification). 2- (1- (tert-Butoxycarbonylamino) cyclobutyl) -2- (4- (((trans) -2- (hydroxymethyl) cyclopropyl) uta-1,3-diinyl) -benzamido) (S) -methyl acetate ( Crude 550mg, l.lmmol) was dissolved in dichloromethane (6mL) and treated with 6mL of trifluoroacetic acid for 10 minutes at room temperature. The volatiles were removed and the residue was re-evaporated from dichloromethane to give 2- (1-aminocyclobutyl) -2- (4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) -benzamido) (S) -methyl acetate as its trifluoroacetate salt, which was used without further purification. trifluoroacetate product amino (methyl) butyl) -2- (4- (((trans) -2- (hydroxythnethyl) cyclo-propyl) buta-l, 3 -diinyl) benzamido) acetate (S) -methyl crude obtained in the above reaction was dissolved in isopropanol (2mL) and treated at 4 ° C for 20 hours with 50% aqueous hydroxylamine (2mL). The volatiles were removed and the residue dissolved in water / acetonitrile in acidification with trifluoroacetic acid. The material was purified by reverse phase HPLC (column 2", 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 2%) using the following gradient: 2% lOmin 2-15 5min 15-45 300min The product eluted between 58 and 78 minutes, and the desired fractions were lyophilized to obtain N - ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide as its trifluoroacetate salt (white solid, 120mg, 0.24mmol, 28% 4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta-l, 3-diinyl) benzoic). Mass spectral data: expected (M + l): 382.4, observed: 382.1. Proton NMR (400MHz, dmso-d6): 11.3 (br s, 1H), 9.24 (s, 1H), 8.62 (d, 1H, J = 8.8Hz), 8.17 (br s, 3H), 7.86 (d, 2H, J = 8.4Hz), 7.63 (d, 2H, J = 8.4Hz), 4.92 (d, 1H, J = 9.2Hz), 4.69 (t, 1H, J = 5.8Hz), 3.40 (m, 1H) , 3.26 (m, 1H), 2.12-2.21 (m, 4H), 1.89 (m, 1H), 1.80 (m, 1H), 1.39-1.46 (m, 2H), 0.84-0.93 (m, 2H). An additional 80mg was obtained in the re-purification of impure fractions.

The fractions containing the majority of N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((cis) -2- (hydroxymethyl) cyclopropyl) buta- 1,3-diinyl) benzamide (an impurity in the main isomer) were lyophilized separately and repurified under the same gradient. The desired fractions were accumulated and lyophilized to give N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((cis) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) -benzamide as its trifluoroacetate salt (3.2mg, 8.4umol, 1% 4- (((trans) -2- (hydroxymethyl) cyclopropyl) buta-1, 3-diinyl) enzoic acid ). Mass spectral data: expected (M + l): 382.4, observed: 382.1. Proton NMR (400MHz, dmso-d6): 11.30 (bs s, 1H), 9.24 (s, 1H), 8.62 (d, 1H, J = 9.2Hz), 8.16 (br s, 3H), 7.86 (d, 2H, J = 7.6Hz), 7.65 (d, 2H, J = 7.6Hz), 4.92 (d, 1H, J = 9.2Hz), 4.70 (t, 1H, J = 5.2Hz), 3.49 (m, 1H) , 3.40 (m, 1H), 2.12-2.22 (m, 4H), 1.82-1.95 (m, 1H), 1.70-1.80 (m, 2H), 1.36 (m, 1H), 1.08 (m, 1H), 0.58 (m, 1H).

N. Synthesis of N- ((S) -1- (1- (dimethylamino) cyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((1, 2-trans) -2- (hydroxymethyl) -cyclopropyl) buta-1,3-diinyl) benzamide (1-19) N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((trans) -2- (hydroxymethyl) -cyclopropyl) buta-1, 3- trifluoroacetate diinyl) benzamide 1-17 (80mg, 0.16mmol, leq) was dissolved in N / N-dimethylformamide (800pL) and treated with paraformaldehyde (48.5mg, l.lmmol, lOeq) and N, N-diisopropylethylamine (56L, 0.32mmol, 2eq) at room temperature for 21 hours. Sodium cyanoborohydride (30mg, 0.48mmol, 3eq) was added followed by methanol (800pL) and acetic acid (28L, 0.48mmol, 3eq). The mixture was kept at room temperature for 3 days, then more sodium cyanoborohydride and acetic acid (28μ? _) Was added. After an additional day at room temperature, another 28 pL of acetic acid was added. After a further day at room temperature the material was purified by reverse phase HPLC (1"column, 20mL / min, 0.1% TFA in water / acetonitrile, equilibrated at 2%) using the following gradient: 2% 5min 2-18% 5min 18-98 80min The desired fractions were accumulated and lyophilized to give N- ((S) -1- (1- (dimethylamino) cyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((1,2-trans) - 2- (hydroxymethyl) cyclopropyl) buta-l (3-diinyl) benzaraide as its trifluoroacetate salt (63mg, 0.12mmol, 74%) Mass spectral data: expected (M + l): 410.5, observed: 410.2. of proton (400MHz, dmso-d6): 11.29 (s, 1H), 9.72 (br s, 1H), 9.30 (s, 1H), 8.95 (d, 1H, J = 9.2Hz), 7.88 (d, 2H, J = 8.4Hz), 7.61 (d, 2H, J = 8.4Hz), 5.05 (d, 1H, J = 9.2Hz), 4.7 (br s, 1H), 3.40 (dd, 1H, J = 5.2, 11.6Hz ), 3.22 (dd, 1H, J = 6.4, 11.2Hz), 2.76 (d, 3H, J = 4.4Hz), 2.68 (d, 3H, J = 4.4Hz), 2.33 - 2.42 (m, 3H), 1.62 - 1.78 (m, 2H), 1.37 - 1.46 (m, 2H), 0.84 - 0.92 (m, 2H). 0. Synthesis of N- ((S) -2- (hydroxyamino) -1- ((S) -morpholin-3-yl) -2-oxoethyl) -4- ((. {1,2-trans) -2- (hydroxymethyl) cyclopropyl) -buta-1,3-diinyl) benzamide (1-20) A triethylamine (3.91 mL, 0. 0281 mol) dropwise at -16 ° C and after 5 min the resulting mixture was treated with isobutyl chloroformate (3.37 mL, 0.02600 mol) slowly at -15 ° C for a duration of 30 min, then the reaction mixture was stirred at -15 ° C ~ 6 ° C for 16h (overnight), the LCMS monitored the reaction process, filtered, washed the solid with ether (30mL x 2), the filtrate was transferred to the next step. The previous filtrate was treated with a solution of diazomethane (0.24 mol) in ether (480 mL) at 0 ~ 7 ° C for 16 h (overnight), then the reaction was quenched with glacial acetic acid (5 mL), water ( 50 mL), and the organic layer was washed with sodium bicarbonate (50 mL x 2), brine (50 mL), the aqueous layer was re-extracted with ether (200 mL x 2), the combined organic layer was dried over Anhydrous sodium sulfate, filtered, concentrated to give the crude residue which was purified by silica gel chromatography (20% ethyl acetate in petroleum as eluent), gave (R) -tert-butyl-3- (2 -diazoacetyl) morpholine-4-carboxylate 2 as a yellow solid, 4.56g (87% purity, LCMS, 254nm), yield: 77%, 2 steps.

A pre-cooled suspension of (R) -ter-butyl-3- (2-diazoacetyl) morpholine-4-carboxylate 2 (4.55 g, 17,824 mmol) and silver benzoate (25 mg, 0.11 mmol) in methanol (30 mL ) triethylamine (250 L, 1.78 mmol) was added dropwise and the suspension was stirred at 25 ° C for 16 h in the dark (overnight, oil bath), then filtered, the filtrate was evaporated to dryness and the The residue was purified by silica gel chromatography (10% ethyl acetate in petroleum as eluent), gave (S) -tert-butyl-3- (2-methoxy-2-oxoethyl) morpholine-4-carboxylate 3 as a colorless solid, 3.34 g, yield: 78%.

To a pre-cooled solution of (S) -ter-butyl-3- (2-methoxy-2-oxoethyl) morpholine-4-carboxylate 3 (2737 g, 0.0106 mol) in THF (100 mL) was added NaHMDS (8.15 mL, 0.0159 mol) slowly at -100 ° C and the resulting suspension was stirred at -100 ° C for 1 h, then a solution of trisilyl azide (5.99 g, 0.0170 mol) in THF (10 mL) was added slowly (keep the temperature inside below -100 ° C), and additional lh stirring at -100 ° C was continued. Glacial acetic acid (3.18 g, 0.0530 mol) was added rapidly to quench the reaction, and the reaction mixture was allowed to warm to room temperature, stirred for 3 h, finally diluted with DCM (200mL x 2) and washed with NaHCO3. saturated (lOOmL), brine (lOOmL) and water (lOOmL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated to give the crude residue which was purified by reverse phase chromatography (Gradient: 10% acetonitrile in water for 5min, increases to 30% acetonitrile in water within 3min, 30% acetonitrile in water for 3min, 30-45% acetonitrile in water, 3min, 45% acetonitrile in water for 5min, flow rate: 30mL / min), to give (S) - ter- butyl-3 - (1-azido-2-methoxy-2-oxoethyl) morpholine-4-carboxylate 4 (mixture of two diastereoisomers) as a white solid, 1.22 g, yield: 38% S68-A, 170mg SM68-B, 230mg SM68-C, 30mg A mixture of (S) -tert-butyl-3- (l-azido-2-methoxy-2-oxoethyl) morpholine-4-carboxylate 4 (1.2 g, 4 mmol), Pd (OH) 2 / C (200 ' mg, 15% by weight), 2M HC1 (2 mL, 4 mmol) in EtOAc / MeOH (20 mL / 3 mL) was degassed and treated with hydrogen gas at 30 ° C under 1 atmosphere for 16 h. The mixture was filtered, and the solid was washed with EtOAc (20mL). The combined organics were washed with water (20mL), saturated NaHC03 (20mL, to adjust the pH = 8), then dried over anhydrous sodium sulfate, filtered, concentrated to give the crude residue which was purified by flash chromatography. reverse phase (Gradient: 10% acetonitrile in water for 5 min, increase to 30% acetonitrile in water within 3 min, 30% acetonitrile in water for 3 min, 30-45% acetonitrile in water, 3 min, acetonitrile 45% in water for 5min; Flow rate: 30mL / min). The purified residue was then sub-atomized to the quital separation, yielding (S) -tert-butyl-3- ((S) -l-amino-2-methoxy-2-oxoethyl) morpholine-4-carboxylate (SM68-A, 170 mg , yellow oil); (S) -ter-butyl-3- ((R) -l-amino-2-methoxy-2-oxoethyl) morpholine-4-carboxylato (SM68-B, 230 mg, yellow solid); and SM68-C, 30 mg, yellow oil, total yield: 41%. 4- (((trans) -2- (hydroxymethyl) cyclopropyl) -buta-1,3-diinyl) benzoic acid (140 mg, 0.58 mmol) and (S) -tert-butyl-3- (() were dissolved. S) -l-amino-2-methoxy-2-oxoethyl) -morpholine-4-carboxylate (160 mg, 0.58 mmol) in DMF (2 mL). W, N-diisopropylethylamine (255 L, 1.46 mmol) was added followed by HATU (266 mg, 0.70 mmol). The reaction was maintained at room temperature for 30 minutes, then it was partitioned between 1M citric acid and ethyl acetate. The products Organic washes with acid. 1M citric acid, saturated sodium bicarbonate then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. 380mg of (S) -ter-butyl-3- ((S) -1- (4- (((1, 2-trans) -2- (hydroxymethyl) -cyclopropyl) buta-l, 3-diinyl) enzymido) Crude 2-methoxy-2-oxoethyl) -morpholine-4-carboxylate was obtained.

(S) -tert-butyl-3- ((S) -1- (4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamido) - was dissolved. 2-methoxy-2-oxoethyl) morpholine ^ 4-carboxylate (crude, 380 mg) in dichloromethane (4 mL), se. added trifluoroacetic acid (4 mL). After 5 minutes at room temperature the mixture was diluted with 10 mL of dichloromethane and evaporated. 2, 2, 2-Trifluoroacetate (S) -methyl-2- (4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) venza-mido) - 2- ((S) -morpholine-3-yl) acetate thus obtained was used crude in the next step. 2, 2, 2-Trifluoroacetate of (S) -methyl-2- (4- (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) -benzamido) -2 - ((S) -morpholine-3-yl) acetate (crude from the previous reaction) was dissolved in isopropanol (1 mL), and aqueous hydroxylamine (50%, 1 mL) was added. The solution was kept at 4 ° C for 18 hours then evaporated. Water was added and the mixture was acidified with TFA. The crude material was purified by RP HPLC (2"column, 50mL / min, 0.1% TFA in water / ACN, balanced @ 2%): 2% lOmin 2-11% 5min 11-41% 300min The desired compound was eluted at 88-99 minutes. The desired fractions were lyophilized to give N - ((S) -2- (hydroxyamino) -1- ((S) -morpholin-3-yl) -2-oxoethyl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide 1-20 as its trifluoroacetate salt (white solid, 118 mg, 0.30 mmol, 52% of 4- (((trans) -2- ( hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzoic acid). Mass spectral data: expected (+ l): 398.2, observed: 398.1. Proton NMR (400MHz, dmso-d6): 11.17 (s, 1H), 9.14 (s, 1H), 9.08 (br s, 1H), 8.93 (br s, 1H), 8.83 (d, 1H, J = 8.4 Hz), 7.89 (d, 2H, J = 8.0Hz), 7.62 (d, 2H, J = 8.0Hz), 4.69 (br s, 1H), 4.63 (t, 1H, J = 8.8HZ), 3.83 - 3.86 (m, 2H), 3.60 - 3.66 (m, 2H), 3.50 (t, 1H, J = 11.0Hz), 3.39 (m, 1H), 3.20 - 3.23 (m, 2H), 3. 05 (m, 1H), 1.39-1.45 (m, 2H), 0.87-0.92 (m, 2H).

P. Synthesis of N- ((S) -2- (hydroxyamino) -1- ((S) -4-methyl-morpholin-3-yl) -2-oxoethyl) -4- (((1, 2-trans ) -2- (hydroxymethyl) -cyclopropyl) buta-1,3-diinyl) benzamide (1-21) N- ((S) -2- (hydroxyamino) -1- ((S) -morpholin-3-yl) -2-oxoethyl) -4- (((1, 2-trans) -2- (hydroxymethyl) trifluoroacetate ) cyclopropyl) buta-1,3-diinyl) benzamide 1-20 (10 mg, 20 μm) was dissolved in DMF (100 pL). Paraformaldehyde (5.9 mg, 200 mol) was added followed by N, N-diisopropylethylamine (6.8 L, 39 pmol). The mixture was stirred at room temperature for 19 hours. Sodium cyanoborohydride (3.7 mg, 59 μp), methanol (100 μ ?.) and acetic acid (4.5 μL, 78 μ ??) were added and the mixture was stirred at room temperature for an additional 3 days . The mixture was purified by RP HPLC (2"column, 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 2%): 2% lOmin 2-15% 5min 15-95% 80min Product eluted between 23 and 24 minutes. The desired fractions were lyophilized to give N- ((S) -2- (hydroxyamino) -1- ((S) -4-methylmorpholin-3-yl) -2-oxoethyl) -4- (((1,2- trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide as its trifluoroacetate salt (7.0 mg, 13 pmol, 68%). Mass spectral data: expected (M + l): 411.2, observed: 412.2.

Q. Synthesis of N- ((S) -3-amino-1-hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- ((E) -4- (1, 2-trans) - 2- (hydroxymethyl) cyclopropyl) but-l-ene-3-ynyl) benzamide (1-22) 3-Amino-2- (4- (E) -4- (1S, 2S) -2- (hydroxymethyl) cyclopropyl) but-l-en-3-ynyl) benzamido) -3-methylbutane-ato of ( S) -methyl (400 mg, 1.08 mmol, 1.0 equivalents) to a round bottom flask. To this was added isopropyl alcohol (2.0 mL), the solution was formed in a vortex for 2 minutes, the flask was cooled in an ice bath, then a solution of hydroxylamine (1.4 mL, 21.60 mmol, 20 equivalents) was added and the flask was placed at 4 ° C for about 48h. The reaction was concentrated, (at 0 ° C) to remove the isopropyl alcohol, and then acidified at 0 ° C using TFA (3 mL). Additional water (15 mL) and ACN (3 mL) were added. The product was purified by reverse phase HPLC (2"column, 50mL / min, 0.1% TFA in water / ACN, equilibrated @ 2% B) .The column was loaded at 10mL / min, 2¾B using syringe filter ( 24 mL) and then increased to 50 mL / min for 1 minute.A gradient from 2% B to 95% B was run for 73 min.The desired fractions were combined, frozen and placed in a lyophilizer. mg of N- ((S) -3-amino-l- (hydroxy-amino) -3-methyl-l-oxobutan-2-yl) -4- ((E) -4- ((1, 2-trans ) 2- (hydroxymethyl) cyclopropyl) but-l-en-3-ynyl) benzamide, 1-22, TFA LCMS M + l expected = 372.2, observed = 372.2 XH NMR (DMSO-d6) - 0.77-0.81 (m, 2H), 1.25 (s, 3H), 1.30 (s, 3H), 1.34-1.37 (m, 1H), 3.20-3.24 (dd, 1H), 3.37-3.41 (dd, 1H), 4.65-4.67 (d, 1H), 6.43-6.47 (d, 1H), 6.87-6.91 (d, 1H), 7.55-7.57 (d, 2H), 7.84-7.86 (d, 2H), 7.99 (br, 2H), 8.38 -8.41 (d, lH), 9.22 (br, 1H), 11.20 (s, 1H).

R. Antimicrobial activity Bacterial tests and crops Bacterial isolates of concentrated solutions frozen at -70 ° C were grown by overnight passages at 35 ° C in room air on Mueller-Hinton agar (Becton Dickinson, Franklin Lakes, NJ). The clinical isolates tested were obtained from several geographically diverse hospitals, in the United States and abroad (Focus Diagnostics, Herndon, VA and JMI, North Liberty, IA). The quality control strains were from the American Type Culture Collection (ATCC, Rockville, MD).

Susceptibility tests The minimum inhibitory concentrations (MIC) were determined by the broth microdilution method according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). In summary, organism suspensions were adjusted to a 0.5 McFarland standard to produce a final inoculum between 3xl05 and 7xl05 colony forming units (CFU) / mi. Dilutions of drug and inoculums were made in Mueller-Hinton broth adjusted with cations, sterile (Beckton Dickinson). An inoculum volume of 100 ml was added to the concavities containing 100 ml of broth with 2-fold serial dilutions of drug. All inoculated microdilution trays were incubated in ambient air at 35 ° C for 18-24 h. After incubation, the lowest concentration of the drug that prevented visible growth (OD600 nm <0.05) was recorded as the MIC. The performance of the assay was monitored by the use of laboratory quality control strains and levofloxacin, a compound with a defined MIC spectrum, in accordance with the CLSI guidelines. Typically, the compounds of the present invention have MIC values of 0.03-16 g / ml. For this purpose, the data for certain representative compounds are shown in Table II below.

Table II Minimum Inhibitory Concentrations (MIC) Key of MIC: A = MIC 2.0 g / ml or less B = MIC greater than 2.0 g / ml at 16.0 pg / ml C = MIC greater than 16.0 g / mL * AECO001 is E. coli ATCC25922; APAE001 is Pseudomonas aeruginosa ATCC27853; AKPN001 is Klebsiella pneumoniae ATCC43816 APAE002 is a clinical isolate of Pseudomonas aeruginosa that expresses a normal level of effluvium activity.

S. Tolerability In Vivo Each compound was administered to the mice by subcutaneous injection. The mice were housed in a group (five per cage) at 18-280C and -50% humidity, and were fed normal rodent food. Water and food were given ad libitum. The mice were given subcutaneous doses of no more than 20 ml per kg of body weight.

Groups of three mice were dosed at 50, 100, 200, 400 or 600 mg / kg / day in a single dose of the test compound by subcutaneous injection in a formulation consisting of 15% Captisol in 20 mM acetate buffer, pH 5. Subcutaneous injections were made in the back in the interscapular area. In this area volumes of 10-20 ml / kg were injected. The needle was inserted parallel to the surface of the skin to a depth such that the point of the needle is within a subcutaneous cavity. A gentle but firm pressure on the plunger was used to eject the contents of the syringe.

Observations were taken at a variety of time points after the dose: 30 seconds - 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 75 minutes; 9.0min, 105min, 2 hours, and every hour thereafter until the animals showed signs of recovery near the baseline, or up to 4 hours after the dose, the first thing that occurs. The minimum time interval for monitoring the mice was 30 minutes after the dose if the animals were alert, normal and sensitive. If the animals showed toxicity effects, they were monitored closely until they showed signs of recovery near baseline, or up to 4 hours after the dose, the first thing that occurs. The animals were maintained for 72 hours after dosing for clinical observation, including monitoring of survival and activity level. The observations considered symptoms of affliction of the central nervous system (such as, attack, lethargy, lying position, immobility, hyperactivity), neuromuscular abnormalities (such as ataxia, twitching, convulsions, extended extremities, jumping or kicking), autonomic symptoms (such as salivation, tearing, urination, defecation, piloerection or strabismus), respiratory distress (such as difficult or rapid breathing, depression, panting or racing), stereotypical behaviors (such as repetitive chewing, circling, stimulation, grooming, sniffing, head movements or stooped posture) and abnormal behaviors such as wet dog escape or shake behavior.

A known compound, No_alc (the synthesis and activity of which is described in International PCT Publication No. 2008/06676; compound 91-12), showed a maximum tolerated dose in mice of less than 50 mg / kg when injected subcutaneously into mice, while a dose of 30 mg / kg is required per subcutaneous injection as a static dose against K. pneumonia in mice. Compound 1-1 of the present invention, which differs from this known compound, simply by a hydroxymethyl substitution of the cyclopropyl group, has antimicrobial activity comparable to that of No_alc but is substantially better tolerated in mammals (maximum tolerated dose of about 200 mg / kg when injected subcutaneously in mice).

\ Table III provides the results of the observations of the subcutaneously dosed mice. Tolerability was characterized by clinical observation. Class A was assigned to animals with few symptoms of toxicity such as occasional short pause when moving, slight forced breathing, or slight lethargy with rapid recovery (for example, within 10 min). Class B was assigned to animals that show some symptoms of toxicity, such as increased pause when moving, slight lethargy with a longer recovery time (up to 1 hour) during which the animals are still able to move. Class C was assigned to animals with moderate to severe symptoms of toxicity, such as lethargy, lying down, lethargy with strabismus and forced breathing, severe twitching (jumping, kicking), or escape behavior. Finally, class D was assigned when some of the lethal effects (including moribund state requiring euthanasia) occurred within the clinical observation period (up to 72 hours after the dose).

Table III In vivo Tolerability T. Synergy with a Second Antibacterial Agent Bacterial isolates of concentrated solutions frozen at -70 ° C of K. pneumoniae (ATCC 43816) were grown by overnight passages at 35 ° C in Mueller-Hinton agar ambient air (Becton Dickinson, Franklin Lakes, NJ). The minimum inhibitory concentrations (MIC) were determined by the broth microdilution method according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI). In summary, organism suspensions were adjusted to a 0.5 McFarland standard to produce a final inoculum between 3xl05 and 7x10s colony forming units (CFU) / ml. The dilutions of drugs and inoculums were made in Mueller-Hinton broth adjusted with cations, sterile (Beckton Dickinson). An inoculum volume of 100 ml was added to the concavities containing 100 ml of broth with serial dilutions of 2 times the drug. All inoculated microdilution trays were incubated in ambient air at 35 ° C for 18-24 hours. After incubation, the lowest concentration of the drug that prevented the visible growth (OD60o nm <0.05) was recorded as the MIC.

Normal chessboard type tests were performed with a combination of the indicated agents and (I- N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 - (((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-di-inyl) benzamide (1-1) 1). Table IV provides the FICI calculated according to the normal techniques. Compound I-1 was synergistic in vitro with vancomycin, teicoplanin, erythromycin, azithromycin, rifampicin and novobiocin.

.Table IV FICI The in vivo synergy of N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- ( hydroxymethyl) cyclopropyl) buta-1,3-diinyl) benzamide (1-1) with vancomycin was examined in model in vivo deficiency of neutropenic thigh. The model was run essentially as described by Craig et al. (See Gudmundsson et al., "Murine Thigh Infection Model", Handbook of Animal Models of infection, MA Sande and O. Zak, Eds., London: Academic Press, 1999, pages 137-144). Briefly, the mice became neutropenic before infection with 2 doses of cyclophosphamide, and then were infected intramuscularly in the thigh with inoculations of 103-105 CFU of K. pneumo. (ATCC 43816). Antibiotics or vehicles were administered only as a negative control twice in 2 hours and 14 hours after infection. The animals remained neutropenic for the duration of the experiment in order to minimize the effect of the white blood cells on the infection such that the microbial reading measures the in vivo interaction of the drugs and bacteria. Twenty-four hours after infection, the thighs were harvested, homogenized, and plated to measure the number of CFUs surviving per thigh. The thighs of a subset of animals were also collected 2 hours after infection to record the CFU present just before the first antibiotic treatment (pre-treatment). The static dose, defined as the dose required to result in a 24-hour CFU load that is identical to that measured at 0 hours after infection, was calculated by normal methods in Prizm (GraphPad Software) of a response curve of dose.

The purpose of these studies was to quantitatively assess whether the combination of 1-1 with a candidate synergistic agent gives a greater reduction. in the accounts in this model of efficiency in vivo than what the sum of each agent alone does.

Vancomycin exhibits significant in vivo synergy with LpxC inhibitors for the treatment of K. pneumo infections. ATCC43816. As indicated in Figure 1, treatment of mice infected with vancomycin alone at 220 mg / kg / day resulted in a significant reduction in CFU. However, when co-dosed with compound 1-1 the static doses of the LpxC inhibitors were significantly reduced (Figure 1 and Table V).

Table V Taken together, these data show that vancomycin demonstrates surprising in vivo synergy with N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ((( 1, 2-trans) -2- (hydroxymethyl) cyclopropyl) buta-1,3-di-inyl) -enoxamide (1-1). it should be understood that the organic compounds according to the invention may exhibit the phenomenon of tautomerism. Since the chemical structures within this specification can only represent one of the possible tautomeric forms, it should be understood that the invention encompasses any tautomeric form of the drawn structure.

Additionally, as long as particular embodiments of the present invention have been shown and described for purposes of illustration, it will be understood, of course, that the invention is not limited thereto since modifications can be made by persons skilled in the art. technique, particularly in view of the above teachings, without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except by the appended claims.

All United States patents, publications of United States patent applications, United States patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification are incorporated herein by reference, in its entirety to the degree not inconsistent with the present description.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (23)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A compound of the formula I: or a stereoisomer or pharmaceutically acceptable salt thereof, characterized in that A is a C3-C6-substituted cycloalkyl, wherein at least one substituent is a primary C1-C3 alcohol; B is absent, -CH = CH-, -C = C- or an unsubstituted phenyl; C is -CH = CH-, -C = C- or an unsubstituted phenyl, where if B is -CH = CH-, then C is also not -CH = CH-; R1, R2 and R3 are independently selected from hydrogen and C1-C3-substituted or unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached, form an unsubstituted C3-C6-cycloalkyl group, or R2 and R3, together with the carbon atom and Q to which they are attached, form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 ring atoms, wherein 1-2 heterocyclic ring atoms are selected from N, O and S; Y Q is O or N, wherein R is hydrogen or a Ci-C3-5 unsubstituted alkyl.

2. A compound according to claim 1, characterized in that Q is NR.

3. A compound according to claim 1 or 2, characterized in that R1, R2, and R3 are 10 independently select from hydrogen and Ci-C-substituted or unsubstituted alkyl.

4. A compound according to any of claims 1-3, characterized in that R1, R2, and R3 are independently selected from hydrogen and C1-C3 -alkyl ^ 5 unsubstituted.

5. A compound according to any of claims 1-4, characterized in that R1 and R2 independently are Ci-C3-unsubstituted alkyl.

6. A compound according to any of claims 1-5, characterized in that A is a C3-C6-cycloalkyl substituted with hydroxymethyl.

7. A compound according to any of claims 1-6, characterized in that B is -C = C- and C is -C = C-.

8. A compound according to claim 1, characterized in that it is: N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta -1,3-diinyl) -benzamide (1-1); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((IR, 2R) -2- (hydroxymethyl) cyclopropyl) buta-1 , 3-diinyl) benzamide (1-2); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2S) -2- (hydroxymethyl) cyclopropyl) buta-1 , 3-diinyl) -benzamide (1-3); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-cis) -2- (hydroxymethyl) cyclopropyl) buta -1, 3-diinyl) -benzamide (1-4); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4 '- (((1,2-trans) -2- (hydroxymethyl) -cyclopropyl) ethynyl) biphenyl-4-carboxamide (1-5); N- ((S) -1- (hydroxyamino) -3-methyl-3- (methylamino) -1-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) ) buta-1,3-diinyl) benzamide (1-6); N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) ) cyclopropyl) -buta-1,3-diinyl) benzamide (1-7); N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4 - (((1, 2-trans) -2- (hydroxymethyl) ) cyclopropyl) -buta-1,3-diinyl) benzamide (1-8); N- ((S) -3- (dimethylamino) -1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) ) buta-1,3-diinyl) benzamide (1-9); N- ((S) -3-hydroxy-1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) - buta-1,3-diinyl) benzamide (1-10); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1S, 2R) -2- (hydroxymethyl) -2-methylcyclopropyl) buta-1, 3-diinyl) benzamide (1-11); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-cis) -3- (hydroxymethyl) -cyclobutyl) buta-1, 3-diinyl) -benzamide (1-12); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-trans) -3- (hydroxymethyl) cyclobutyl) buta -1, 3-diinyl) -benzamide (1-13); N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- (((1,3-trans) -3- (hydroxymethyl) cyclopentyl) buta -1, 3-diinyl) -benzamide (1-14); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,4-cis) -4- (hydroxymethyl) cyclohexyl) buta -1, 3-diinyl) -benzamide (1-15); N- (. (S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,4-trans) -4- (hydroxymethyl) cyclohexyl) buta-1, 3-diinyl) -benzamide (1-16); N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxy-amino) -2-oxoethyl) -4- (((trans) -2- (hydroxymethyl) -cyclopropyl) -buta-1-trifluoroacetate , 3-diinyl) -benzamide (1-17); N- ((S) -1- (1-aminocyclobutyl) -2- (hydroxy-amino) -2-oxoethyl) -4- (((cis) -2- (hydroxymethyl) cyclopropyl) -buta-1-trifluoroacetate, 3-diinyl) benzamide (1-18); N- ((S) -1- (1- (dimethylamino) cyclobutyl) -2- (hydroxyamino) -2-oxoethyl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta- 1,3-diinyl) benzamide (1-19); N- ((S) -2- (hydroxyamino) -1- ((S) -morpholin-3-yl) -2-oxoethyl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) ) buta-1,3-diinyl) -benzamide (1-20); N- ((S) -2- (hydroxyamino) -1- ((S) -4-methylmorpholin-3-yl) -2-oxoethyl) -4- (((1,2-trans) -2- (hydroxymethyl) ) cyclopropyl) buta-1,3-diinyl) benzamide (1-21); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ((E) -4- ((1, 2-trans) -2- (hydroxymethyl) -cyclopropyl) but-1-en-3-ynyl) benzamide (1-22); or N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- ((E) -4- ((1, 2-trans) -2- (hydroxymethyl) cyclopropyl) but-1-en-3-ynyl) benzamide (1-23).

9. A compound according to claim 8, characterized in that it is: N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta -1, 3-diinyl) -benzamide (1-1); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 '- (((1,2-trans) -2- (hydroxymethyl) -cyclopropyl) ethinyl) biphenyl-4-carboxamide (1-5); N- ((S) -1- (hydroxyamino) -3-methyl-3- (methylamino) -1-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) - cyclopropyl) uta-1,3-diinyl) benzamide (1-6); N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) ) -cyclopropyl) -buta-1,3-diinyl) benzamide (1-7); N- ((S) -1- (hydroxyamino) -3- (2-hydroxy-ethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) -buta-1,3-diinyl) benzamide (1-8); N- ((S) -3- (dimethylamino) -1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) ) buta-1,3-diinyl) benzamide (1-9); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,3-cis) -3- (hydroxymethyl) -cyclobutyl) buta-1, 3-diinyl) -benzamide (1-12); N- ((S) -3-amino-1- (hydroxyamino) -3-methyl-1-oxobutan-2-yl) -4- (((1,3-trans) -3- (hydroxymethyl) cyclobutyl) - buta-1, 3-diinyl) -benzamide (1-13); or N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 - (((1,3-trans) -3- (hydroxymethyl) -cyclopentyl) uta-1,3-diinyl) -benzamide (1-14).

10. A compound in accordance with Claim 9, characterized in that it is: N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) cyclopropyl) buta -1, 3-diinyl) -benzamide (1-1); N- ((S) -3-amino-l- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4 · '- (((1,2-trans) -2- (hydroxymethyl) - cyclopropyl) ethynyl) biphenyl-4-carboxamide (1-5); N- ((S) -1- (hydroxyamino) -3-methyl-3- (methylamino) -1-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) - cyclopropyl) uta-1,3-diinyl) benzamide (1-6); N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) ) -cyclopropyl) -buta-1,3-diinyl) enzamide (1-7); N- ((S) -1- (hydroxyamino) -3- (2-hydroxyethylamino) -3-methyl-l-oxobutan-2-yl) -4- (((1,2-trans) -2- (hydroxymethyl) ) -cyclopropyl) -buta-1,3-diinyl) benzamide (1-8); or N- ((S) -3- (dimethylamino) -1- (hydroxyamino) -3-methyl-l-oxobutan-2-yl) -4- (((1, 2-trans) -2- (hydroxymethyl) - cyclopropyl) buta-1,3-diinyl) benzamide (1-9).

11. A pharmaceutical composition, characterized in that it comprises a compound according to any of claims 1-10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

12. A method to treat a subject that has a bacterial infection, characterized in that it comprises administering to a subject in need thereof a therapeutically effective amount of a compound according to any of claims 1-10, or a pharmaceutical composition according to claim 11.

13. A method according to claim 11, characterized in that the bacterial infection is an infection of gratnnegative bacteria.

14. A method according to claim 13, characterized in that the infection of Gram-negative bacteria is Pseudomonas aeruginosa, Burkholderia, Enterobacteriaceae, Franciscellaceae, Serratia, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providencia, Yersinia, Morganella or Escherichia coli.

15. A method in accordance with the claim 14, characterized in that the infection of Gram-negative bacteria is Pseudomonas aeruginosa, Burkholderia, Franciscellaceae, Enterobacter, Yersinia or Escherichia coli.

16. A method according to claim 15, characterized in that the infection of Gram-negative bacteria is Pseudomonas aeruginosa.

17. A method in accordance with the claim 15, characterized in that the infection of Gram-negative bacteria is Escherichia coli.

18. A pharmaceutical composition, characterized in that it comprises a second antibacterial agent composed of the formula I: or a stereoisomer or pharmaceutically acceptable salt thereof, wherein A is a substituted C3-C6-cycloalkyl, wherein at least one substituent is a primary Ci-C3 alcohol; B is absent, -CH = CH-, -C = C- or an unsubstituted phenyl; C is -CH = CH-, -C = C- or an unsubstituted phenyl, where if B is -CH = CH-, then C is also not -CH = CH-; R1, R2 and R3 are independently selected from hydrogen and C1-C3-substituted or unsubstituted alkyl, or R1 and R2, together with the carbon atom to which they are attached, form an unsubstituted C3-C3-cycloalkyl group, or R2 and R3, together with the carbon atom and Q to which they are attached, form a substituted or unsubstituted heterocyclic ring, having from 5 to 8 ring atoms, wherein 1-2 heterocyclic ring atoms are selected from N, O and S; Y Q is O or NR, where R is hydrogen or a C ^ -C ^ - unsubstituted alkyl.

19. A pharmaceutical composition according to claim 18, characterized in that the antibacterial agent is vancomycin or rifampin.

20. A pharmaceutical composition according to claim 18, characterized in that the combination demonstrates synergy in vivo.

21. A method for inhibiting a deacetylase enzyme in a gram-negative bacterium, characterized in that it comprises administering to a patient in need of this inhibition a compound according to any of claims 1-10, or a pharmaceutical composition according to claim 11.

22. A method according to claim 21, characterized in that the gram-negative bacterium is Pseudomonas aeruginosa, Burkholderia, Enterobacteriaceae, Franciscellaceae, Serratia, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providencia, Yersinia, Morganella or Escherichia coli.

23. A method for inhibiting LpxC in a gram-negative bacterium, characterized in that it comprises administering to a patient in need of this inhibition a compound according to any of claims 1-10, or a pharmaceutical composition according to claim 11.