WO2020206381A1 - Cephem compounds with latent reactive groups and methods of using and making same - Google Patents

Cephem compounds with latent reactive groups and methods of using and making same Download PDF

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WO2020206381A1
WO2020206381A1 PCT/US2020/026753 US2020026753W WO2020206381A1 WO 2020206381 A1 WO2020206381 A1 WO 2020206381A1 US 2020026753 W US2020026753 W US 2020026753W WO 2020206381 A1 WO2020206381 A1 WO 2020206381A1
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group
compound
substituted
formula
mmol
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PCT/US2020/026753
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French (fr)
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Larry D. SUTTON
Patrick Bureau
Laura MAMANI LAPARRA
Marc Vidal
Simon Woo
Nancy Zhou
Richard M. Keenan
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Sutton Larry D
Patrick Bureau
Mamani Laparra Laura
Marc Vidal
Simon Woo
Nancy Zhou
Keenan Richard M
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Application filed by Sutton Larry D, Patrick Bureau, Mamani Laparra Laura, Marc Vidal, Simon Woo, Nancy Zhou, Keenan Richard M filed Critical Sutton Larry D
Publication of WO2020206381A1 publication Critical patent/WO2020206381A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D477/00—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring
    • C07D477/10—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2
    • C07D477/12—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2 with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached in position 6
    • C07D477/14—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2 with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached in position 6 with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D501/14—Compounds having a nitrogen atom directly attached in position 7
    • C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
    • C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D501/14—Compounds having a nitrogen atom directly attached in position 7
    • C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
    • C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
    • C07D501/24—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids with hydrocarbon radicals, substituted by hetero atoms or hetero rings, attached in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D501/14—Compounds having a nitrogen atom directly attached in position 7
    • C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
    • C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
    • C07D501/24—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids with hydrocarbon radicals, substituted by hetero atoms or hetero rings, attached in position 3
    • C07D501/38—Methylene radicals, substituted by nitrogen atoms; Lactams thereof with the 2-carboxyl group; Methylene radicals substituted by nitrogen-containing hetero rings attached by the ring nitrogen atom; Quaternary compounds thereof
    • C07D501/46—Methylene radicals, substituted by nitrogen atoms; Lactams thereof with the 2-carboxyl group; Methylene radicals substituted by nitrogen-containing hetero rings attached by the ring nitrogen atom; Quaternary compounds thereof with the 7-amino radical acylated by carboxylic acids containing hetero rings

Definitions

  • Antimicrobial resistance is currently so severe that infecting pathogens are most often described in terms of their resistance to antibiotics. Categories include: usual drug resistance, multi-drug resistance, extreme drug resistance and pan-drug resistance [1 ] The need for new antibiotics which are effective against such resistance is crucial [2, 3-8]
  • Beta-lactam antibiotics are one of the most important and successful classes of antimicrobials for the treatment of human infections caused by bacterial pathogens [9] Bacterial resistance to beta-lactam antibiotics is typically facilitated by production of beta-lactamases, which inactivate beta-lactam antibiotics before they can bind their targets [10-12]
  • Cephalosporins a sub-class of beta-lactam antibiotics, were developed in response to the appearance of early beta-lactamases (penicillinases). Thereafter, bacteria evolved beta-lactamases capable of hydrolyzing these first-generation cephalosporins [13, 14]
  • U.S. Patent Nos. 7,384,928; 7,696,354; 8,883,773; 9,085,589; 9, 145,425; 9,290,515; 9,238,65; and 9,334,289 disclose cephem and/or cephalosporin compounds having a catechol or pseudocatechol group.
  • Each of these patents is incorporated by reference herein in its entirety for the cephem and cephalosporin structures disclosed therein, as well as the disclosed methods of synthesizing cephems and cephalosporins.
  • U.S. Patent No. 9,340,566 discloses cephalosporin antibiotics having a moiety containing a five-member ring with a positively charged N substituted on the cephem ring. This patent is incorporated by reference herein in its entirety for the disclosed cephalosporin structures and methods of synthesizing those compounds.
  • U.S. Patent Nos. 4,218,459; 4,298 741 ; 5,055,463; 5,138,050; and 5,395,931 relate to carbapenem compounds having a 6-amido group and provide some detail of synthesis of such compounds.
  • Each of these patents is incorporated by reference herein in its entirety for the disclosed carbapenem structures and methods of synthesizing those compounds, particularly 6-amido substituted carbapenems.
  • U.S. Patent No. 4,347,355 discloses transpeptidase inhibitors that are beta- lactams of the general structure:
  • R among others, is an acyl group
  • R' is hydrogen, lower alkoxy, lower alkoxyalkyl, lower alkyl, phenylthio or lower alkylmercapto.
  • cephem, penem and monobactam compounds having certain aryl and heteroaryl groups to which a latent reactive group is bonded.
  • the aryl and heteroaryl groups are bonded to the beta-lactam ring and the latent reactive group is conjugated to beta-lactam ring via the aryl or heteroaryl group.
  • the latent reactive group is a leaving group having a positively charged nitrogen, and in certain of these embodiments the latent reactive group contains a vicinal diol or is bonded to an unsubstituted or substituted catechol.
  • the compounds contain a latent reactive group which is released on hydrolysis/cleavage of the lactam ring and is believed to function to inhibit beta- lactamases.
  • the cephems are cephalosporins, cephamycins, carbacephems, and oxacephems.
  • the penems are penems (i.e. , an unsaturated b-lactam with a sulfur atom in the five-member ring), carbapenems (i.e., an unsaturated b-lactam with a carbon atom in the five-member ring) or oxapenems (i.e., an unsaturated b-lactam with an oxygen atom in the five-member ring).
  • Preferred cephems are cephalosporins.
  • Preferred penems are carbapenems.
  • the compounds are monobactams.
  • the compounds are cephalosporins.
  • the compounds are carbapenems.
  • the aryl groups are unsubstituted or substituted phenyl groups.
  • the heteroaryl groups are unsubstituted or substituted single ring heteroaryl groups having one, two, three or four nitrogens in the ring.
  • R is an acylamino group (R1CO-NFI-) or an alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group;
  • M is a divalent cephem, penem or monobactam ring selected from:
  • each R A is independently hydrogen, a C1 -C3 alkyl group or a C1 -C3 alkoxy group
  • R B is hydrogen, a C1 -C3 alkyl group or a C1 -C3-alkoxy group
  • R c is hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, acyloxy-substituted alkyl, a carboxyl protecting group or, when the CO2 group to which R c is attached is negatively charged, a pharmaceutically acceptable cation;
  • Zi and Z2 are independently, S, SO, SO2, 0, or C(R E )2, where each R E is independently hydrogen or a C1 -C3 alkyl group;
  • L is an optional linker which is a substituted or unsubstituted ethylene or a substituted or unsubstituted phenyl group and n is 0, if the linker is absent, or is an integer ranging from 1 to 5, if the linker is present;
  • the B ring is an unsubstituted or substituted phenyl ring or an unsubstituted or substituted 6-member heteroaryl ring having one to four ring nitrogens;
  • R5, R6, R7, Re and R9 are independently selected from hydrogen, halogen, cyano, nitro, C1 -C3 alkyl, C1 -C3 haloalkyl, amino, C1 -C3 alkylamino, and C1 -C3 dialkylamino and -CH2-X, with the exception that when the ring atom to which one of R5-R9 is bonded is a nitrogen, that R5-R9 is not present or is independently selected from hydrogen or C1 -C3 alkyl and at least one of Rs to R9 is -CH2-X, where X is an organic or inorganic leaving group.
  • Optional substitution for L groups includes one or more halogen, nitro, cyano, C1 -C3 alkyl, C1 -C3 alkoxy, or -C02R c groups, where R c is as defined in Formula I .
  • R7 is -CH2-X.
  • the B ring is an unsubstituted phenyl or a mono-nitro-substituted phenyl and R7 is -CH2-X.
  • the B ring is an unsubstituted 6-member heteroaryl ring having one nitrogen and R7 is -CH2-X.
  • X is halogen, a C1-C6 ester, a C1-C6 thioester, a C1-C6 alcohol, a C1-C6 alkylamino, a C1-C6 dialkylamino, a Ci -C6 trialkylammonium, a C1-C6 phosphate ester, a C1-C6 phosphite ester, a C1-C6 sulfate ester, a C1-C6 sulfite ester, a thiol, a sulfenyl, an unsubstituted or substituted phenoxy, an unsubstituted or substituted tosyl or a leaving group having a positively charged nitrogen, such as an unsubstituted or substituted pyridinium group.
  • X is a leaving group having a positively charged nitrogen wherein the leaving group contains a vicinal diol
  • -CH2-X has structure set forth in Formula Xi :
  • a + is a leaving group containing a positively charged nitrogen
  • Rii is selected from hydrogen, hydroxyl, halogen, C1 -C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
  • R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1 -C3 alkyl, unsubstituted or substituted C1 -C3 alkoxy, cyano, and nitro groups or
  • R11 with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
  • linker D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two or three heteroatoms (preferably N, S or 0), where m is 1 or 0 to indicate presence or absence of linker D.
  • Optional substitution for the linker D includes one or more halogen, oxo group, C1 -C3 alkyl group, or C1 -C3 alkoxy group.
  • a + group in Formula Xi is bonded to the indicated phenyl ring through linker D or is a partially or fully unsaturated heterocyclic ring fused to the indicated phenyl ring.
  • the indicated phenyl ring is a catechol ring having two hydroxyl groups substituted on any two adjacent ring carbons.
  • Beta-lactam ring systems of the compounds of Formula I include those of cephems, cephamycins, carbacephems, oxacephems, penems, carbapenems, oxapenems, or monobactams.
  • the compound of the disclosure is a pharmaceutically acceptable salt of the compound of Formula I.
  • the salt of the compound may be formed at the carboxylate of the group at the 2-position of the cephem/penem ring; and/or at a carboxylate and/or or at an amino group in the group at the 7-position (cephem) or 6-position (penem) of the ring; and/or at the positively charged nitrogen of the M group.
  • the compound can also be zwitterionic.
  • the compound of the disclosure is a pharmaceutically acceptable solvate of the compound or salt of Formula I.
  • the compound of the disclosure is a pharmaceutically acceptable hydrate of the compound or salt of Formula I .
  • Zi is S, SO, or SO2. In specific embodiments, Zi is S. In specific embodiments, Zi is CH2 or CHR E , where R E is hydrogen or a C1 -C3 alkyl group. More specifically, R E is methyl.
  • Zi is CH2. In specific embodiments, Zi is O.
  • Z2 is S, SO, or SO2. In specific embodiments, Z2 is S. In specific embodiments, Z2 is CH2 or CHR E , where R E is hydrogen or a C1 -C3 alkyl group. More specifically, R E is methyl.
  • Z2 is CH2. In specific embodiments, Z2 is O.
  • R is an optionally substituted alkyl group.
  • R is a hydroxy substituted alkyl group, a hydroxy substituted alkyl group where the hydroxyl group is protected or a halogen substituted alkyl group. More specifically, R is a hydroxy substituted C1 -C6 alkyl group, where the hydroxyl group is optionally protected. More specifically, R is a halogen substituted C1 -C6 alkyl group. More specifically, R is a fluorine substituted C1 -C6 alkyl group. Yet more specifically, R is a hydroxyl substituted C1 -C3 alkyl group, where the hydroxyl group is optionally protected.
  • the C1 -C6 alkyl or the C1 -C3 alkyl is substituted with a hydroxy or a halogen on the 1 -position of the alkyl group.
  • R is 1 -hydroxyethyl, where the hydroxyl group is optionally protected.
  • R is a 1 -fluoroethyl.
  • R is 1 - hydroxyethyl.
  • R is an acylamino group, particularly an acylamino group of a cephalosporin, cephamycin, carbacephem or oxacephem antibiotic.
  • R is R-i-CO-NH-, where Ri is any of various organic groups, including without limit, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted carbocyclic, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl.
  • R1CONH- is an acylamino group of a known beta-lactam antibiotic. A wide variety of beta-lactam antibiotics is known in the art. Acylamino groups of representative known beta-lactam antibiotics are described hereinafter.
  • Ri is an optionally substituted benzyl group.
  • the compounds of Formula I are cephalosporins, where M is Mi , Zi is S and R B is hydrogen.
  • the compounds of Formula I are cephalosporins, where M is Mi , Zi is S, R is an acylamino group and R B is hydrogen.
  • M is Mi, Zi is S, R B is hydrogen and both of R A are hydrogen.
  • M is Mi , Zi is S, R is an acylamino group, R B is hydrogen and both of R A are hydrogen.
  • both R A are hydrogen.
  • one R A is a C1 -C3 alkyl and the other R A is hydrogen.
  • one R A is a methyl and the other R A is hydrogen.
  • the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and R B is C1 -C3 alkyoxy.
  • the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and R B is C1 -C3 alkyoxy.
  • the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and R B is methoxy.
  • the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and R B is methoxy.
  • both R A are hydrogen.
  • one R A is a C1 -C3 alkyl and the other R A is hydrogen.
  • one R A is a methyl and the other R A is hydrogen.
  • the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CH2 or CFKCFh). In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2, or more specifically Z2 is CFI2 or CH(CH3) and R B is hydrogen. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2, or more specifically Z2 is CFI2 or CH(CH3), R B is hydrogen and R is a C1 -C6 hydroxyalkyl group.
  • the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CFI2 or CH(CH3), R B is hydrogen and R is a 1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, R is an (R)-1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CFI2 or CH(CH3), R B is hydrogen and R is a 1 -hydroxyethyl group. In specific embodiments, R is an (R)-1 -hydroxyethyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CFI2 or CH(CH3), R B is hydrogen and R is an acylamino group.
  • the compounds of Formula I are monobactams where M is M3 and n is 0.
  • compounds of Formula I have a cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have a 5- or 6- member cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have an isoquinoline diol or a quinolone diol group.
  • -CFI2-X has the structure set forth in Formula X2:
  • Rii and R12- R-is are generally as defined in Formula Xi or
  • R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
  • R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
  • R16 is hydrogen or unsubstituted or substituted C1 -C8 alkyl or cycloalkyl
  • Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl
  • R18 is a divalent -(CH2) P - moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and Rie together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds.
  • the phenyl ring at the right of the Formula X2 is substituted with at least two hydroxyl groups on adjacent ring carbons and is bonded to the positively charged N through the R18 moiety or is fused to the 5- or 6-member ring formed by R17 and R18.
  • optional substitution is substitution with one or more C1 -C3 alkyl group, C1 -C3 alkoxy group, hydroxyl, halogen, carboxylate or esters thereof, nitro, or cyano.
  • Specific halogens include chlorine or fluorine.
  • the M group of Formula I is selected from any M group from M1 -M3.
  • the B ring of Formula I is selected from any B ring from B-i-Bse.
  • the M group of Formula I is selected from any M group from M1 -M3 and the B ring of Formula I is selected from any B ring from B1 -B58.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, or Bi4.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, B 14 , B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the M group of Formula I is selected from M3-10, or M3-12, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi , B7, Be, or B14.
  • the M group of Formula I is selected from M3-23, or M3-25, and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi , B7, Be, or Bi4.
  • the compounds provided herein exhibit antibiotic activity against Gram-negative bacteria, Gram-positive bacteria, or both Gram-negative and Gram-positive bacteria. In certain embodiments, the compounds provided herein exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In certain embodiments, the compounds provided herein exhibit antibiotic activity against bacteria which produce various beta-lactamases.
  • the compounds provided herein exhibit antibiotic activity against one or more Enterobactenaceae, including but not limited to Escherichia coir, Klebsiella ; Proteus, Citrobacter; Serratia; and/or Enterobacter.
  • the compounds exhibit antibiotic activity against strains of Klebsiella pneumoniae ; Klebsiella oxytoca ; Proteus mirabilis ; Citrobacter freundir, Serratia marcescens ; Enterobacter aerogenes ; and/or Enterobacter cloacae.
  • the compounds provided herein exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL). In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemase. In some embodiments, the compounds exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL), which produce AmpC beta-lactamases or which produce a carbapenemase.
  • ESBL extended spectrum beta-lactamases
  • compositions comprising one or more of the cephem, penem or monobactam compounds disclosed herein.
  • the compositions are pharmaceutical compositions comprising a pharmaceutically effective amount of the one or more compounds.
  • the pharmaceutical composition comprises a sufficient amount of the one or more compounds to inhibit bacterial growth.
  • the pharmaceutical compositions provided herein comprise a beta-lactamase inhibitor or a beta-lactam antibiotic other than a compound of any one of the formulas herein.
  • the pharmaceutical compositions provided herein contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitor other than a compound of Formula I, one or more beta-lactam antibiotic other than a compound of Formula I, one or more monobactam other than a compound of Formula I, one or more carbapenem other than a compound of Formula I or one or more aminoglycoside antibiotic.
  • Pharmaceutical compositions herein optionally further comprise one or more pharmaceutically acceptable carriers or excipients as are known in the art.
  • pharmaceutical compositions herein are for administration by injection. In certain embodiments, pharmaceutical compositions herein are for oral administration.
  • a subject which may be a human or non-human animal, preferably a mammal, by administering one or more of the compounds or pharmaceutical compositions provided herein.
  • compositions provided herein for use in the prevention or treatment of bacterial infections.
  • the bacterial infection is an infection by a bacterium that is a multi-drug resistant bacterium. In certain embodiments, the bacterial infection is an infection by a bacterium which produces extended spectrum beta- lactamases.
  • FIG. 1 is a graph depicting the mean plasma concentration of GL-332 and the comparators, ceftazidime, and ceftriaxone after IV dosing.
  • the data shows that GL- 332 has pharmacokinetic profile similar to other clinically available cephalosporins with an early elimination rate similar to ceftazidime and a terminal elimination half-life similar to ceftriaxone.
  • FIG. 2 shows an ICso inhibition curve of GL-332 for PBP3.
  • the data shows that GL-332 exhibited a very low ICso of PBP3 at a concentration of 10 ng/mL.
  • FIG. 3 shows the effects of GL-332 on binding to PBP3 and thereby, inhibiting bacterial growth using electrophoresis. As demonstrated in FIG. 3, as the concentration of GL-332 is increased (10 fg/mL to 100 pg/mL), the fluorescent signal decreases for both replicate 1 and replicate 2.
  • FIG. 4 is a graph depicting the survival rate (100% survival, p ⁇ 0.05) after also SC dosing in mice.
  • the data shows that the survival rate for the mice administered the vehicle control rapidly decreased at day 1 ; however, the survival rate for GL-332 (QD and BID) was 100% after day 7, comparable to gentamicin.
  • FIGs. 5A-5G shows plots of Vo (initial rate without GL-332)AA (initial rate with GL332 of hydrolysis nitrocefin by AmpC, KPC-2, CTX-M-15, NDM-1 , IMP-1 , OXA- 24, and OXA-48, respectively and best fit lines to a competitive inhibition model.
  • FIGs. 5A-5G reveal that GL-332 inhibited all of the b-lactamases via a competitive mechanism.
  • FIGs. 6A-6C are graphs depicting the plasma concentration of GL-337 after IV, SC, and mean concentration after dosing at 20 mg/kg, respectively. As demonstrated by FIGs. 6B-6C, GL-337 was rapidly absorbed at 20 mg/kg in mice after subcutaneous dosing. DETAILED DESCRIPTION
  • the cephem GL-332 has been found to have potent bactericidal activity against a broad range of different organisms, including E. coli, K. pneumoniae, and P. aeruginosa. GL-332 was found to inhibit bacterial growth and replication at remarkably low concentrations. The results provided herein further show that GL-332 has a low affinity for plasma proteins and inhibits penicillin-binding proteins (PBP). For example, GL-332 can inhibit bacterial growth as evidenced by the MIC data and it is contemplated that GL-332 exerts its effect predominately by potently binding to PBP3 with an ICso as low as 10 ng/mL.
  • PBP penicillin-binding proteins
  • GL-332 exhibits profound protective effects against bacterial infections in vivo. For example, intravitreal administration of GL-332 to mice infected with P. aeruginosa resulted in a 100% mice survival rate after 7 days. Kinetic studies together with ICso values indicate that GL-332 is a suitable drug candidate for the development of more potent b-lactamase inhibitors, and that it is not susceptible to inactivation by serine b-lactamases from the Ambler class enzymes A-D.
  • GL-332 As superior to previously disclosed antibacterial compounds. Accordingly, provided herein is GL-332 and salts or solvates thereof, as well as pharmaceutical compositions comprising GL-332 or salts or solvates thereof. Also provided herein are additional cephem, penem, and monobactam compounds sharing structural characteristics with GL-332, including salts and solvates of these compounds, as well as pharmaceutical compositions comprising these compounds. Further provided herein are methods of using the compounds and pharmaceutical compositions provided herein to inhibit bacterial growth and/or replication and to treat bacterial infection, and the use of the disclosed compounds and pharmaceutical compositions for these methods. Further provided are kits comprising the compounds and pharmaceutical compositions provided herein.
  • novel cephem, penem and monobactam compounds having (1 ) a phenyl or (2) a heteroaryl moiety, such as a pyridyl group, at the 3-position in the cephem or penem ring or at the N of the monobactam ring, to which a leaving group X is bonded through a methylene group (- CH2-X).
  • the leaving group has a positively charged N atom.
  • the leaving group also contains a vicinal diol or is bonded to an unsubstituted or substituted catechol.
  • the leaving group is a positively charged leaving group.
  • monobactam compounds having a styrenylmethylene moiety at the N of the monobactam ring to which a leaving group is bonded.
  • the leaving group has a positively charged N atom.
  • the leaving group also contains a vicinal diol or is bonded to an unsubstituted or substituted catechol.
  • the compounds provided herein contain a latent reactive group which is released on hydrolysis/cleavage of the lactam ring and is believed to function to inhibit beta-lactamases.
  • the beta-lactams of the invention include cephems, penems and monobactams. More specifically, the cephems are cephalosporins, cephamycins, carbacephems, and oxacephems. More specifically, the penems are penems, carbapenems or oxapenems. Preferred cephems are cephalosporins. Preferred penems are carbapenems.
  • the disclosure relates to cephem and penem compounds substituted with a phenyl or heteroaryl ring which is bonded to a positively charged leaving group and which is in turn bonded to a vicinal diol group, or more specifically a catechol moiety.
  • the positively charged leaving group has a positively charged nitrogen.
  • R is an acylamino group (R1CO-NH-) or an alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group;
  • M is a divalent cephem, penem or monobactam ring selected from:
  • each R A is independently hydrogen, a C1 -C3 alkyl group or a C1 -C3 alkoxy group
  • R B is hydrogen, a C1 -C3 alkyl group or a C1 -C3-alkoxy group
  • R c is hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, acyloxy-substituted alkyl, a carboxyl protecting group or, when the CO2 group to which R c is attached is negatively charged, a pharmaceutically acceptable cation;
  • Zi and Z2 are independently, S, SO, SO2, 0, or C(R E )2, where each R E is independently hydrogen or a C1 -C3 alkyl group;
  • L is an optional linker which is a substituted or unsubstituted ethylene or a substituted or unsubstituted phenyl group and n is 0, if the linker is absent, or is an integer ranging from 1 to 5, if the linker is present;
  • the B ring is an unsubstituted or substituted phenyl ring or an unsubstituted or substituted 6-member heteroaryl ring having one to four ring nitrogens;
  • R5, R6, R7, Re and R9 are independently selected from hydrogen, halogen, cyano, nitro, C1 -C3 alkyl, C1 -C3 haloalkyl, amino, C1 -C3 alkylamino, and C1 -C3 dialkylamino and -CH2-X, with the exception that when the ring atom to which one of R5-R9 is bonded is a nitrogen, that R5-R9 is not present or is independently selected from hydrogen or C1 -C3 alkyl and at least one of Rs to R9 is -CH2-X, where X is an organic or inorganic leaving group.
  • Optional substitutions for L groups includes one or more halogen, nitro, cyano, C1 -C3 alkyl, C1 -C3 alkoxy, or -C02R c groups, where R c is as defined in Formula I .
  • R7 is -CH2-X.
  • the B ring is an unsubstituted phenyl or a mono- nitro-substituted phenyl and R7 is -CH2-X.
  • the B ring is an unsubstituted 6-member heteroaryl ring having one nitrogen and R7 is -CH2-X.
  • X is halogen, a C1-C6 ester, a C1-C6 thioester, a C1-C6 alcohol, a C1-C6 alkylamino, a C1-C6 dialkylamino, a Ci -C6 trialkylammonium, a C1-C6 phosphate ester, a C1-C6 phosphite ester, a C1-C6 sulfate ester, a C1-C6 sulfite ester, a thiol, a sulfenyl, an unsubstituted or substituted phenoxy, an unsubstituted or substituted tosyl or a leaving group having a positively charged nitrogen, such as an unsubstituted or substituted pyridinium group.
  • X is a leaving group having a positively charged nitrogen wherein the leaving group contains a vicinal diol
  • -CH2-X has the structure set forth in Formula Xi:
  • a + is a leaving group containing a positively charged nitrogen
  • Rii is selected from hydrogen, hydroxyl, halogen, C1 -C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
  • R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1 -C3 alkyl, unsubstituted or substituted C1 -C3 alkoxy, cyano, and nitro groups or Rii with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; and
  • linker D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two or three heteroatoms (preferably N, S or 0), where m is 1 or 0 to indicate presence or absence of linker D.
  • Optional substitution for the linker D includes one or more halogen, oxo group, C1 -C3 alkyl group, or C1 -C3 alkoxy group.
  • a + group in Formula Xi is bonded to the indicated phenyl ring through linker D or is a partially or fully unsaturated heterocyclic ring fused to the indicated phenyl ring.
  • the indicated phenyl ring is a catechol ring having two hydroxyl groups substituted on any two adjacent ring carbons.
  • Beta-lactam ring systems of the compounds of Formula I include those of cephems, cephamycins, carbacephems, oxacephems, penems, carbapenems, oxapenems, or monobactams.
  • the compound of the disclosure is a pharmaceutically acceptable salt of the compound of Formula I.
  • the salt of the compound may be formed at the carboxylate of the group at the 2-position of the cephem/penem ring; and/or at a carboxylate and/or or at an amino group in the group at the 7-position (cephem) or 6-position (penem) of the ring; and/or at the positively charged nitrogen of the M group.
  • the compound can also be zwitterionic.
  • the compound of the disclosure is a pharmaceutically acceptable solvate of the compound or salt of Formula I.
  • the compound of the disclosure is a pharmaceutically acceptable hydrate of the compound or salt of Formula I .
  • Zi is S, SO, or SO2. In specific embodiments, Zi is S. In specific embodiments, Zi is Chte or CHR E , where R E is hydrogen or a C1 -C3 alkyl group. More specifically, R E is methyl.
  • Zi is CH2. In specific embodiments, Zi is O.
  • Z2 is S, SO, or SO2. In specific embodiments, Z2 is S. In specific embodiments, Z2 is Ch or CHR E , where R E is hydrogen or a C1 -C3 alkyl group. More specifically, R E is methyl.
  • Z2 is CH2. In specific embodiments, Z2 is O.
  • R is an optionally substituted alkyl group.
  • R is a hydroxy substituted alkyl group, a hydroxy substituted alkyl group where the hydroxyl group is protected or a halogen substituted alkyl group. More specifically, R is a hydroxy substituted C1 -C6 alkyl group, where the hydroxyl group is optionally protected. More specifically, R is a halogen substituted C1 -C6 alkyl group. More specifically, R is a fluorine substituted C1 -C6 alkyl group. Yet more specifically, R is a hydroxyl substituted C1 -C3 alkyl group, where the hydroxyl group is optionally protected.
  • the C1 -C6 alkyl or the C1 -C3 alkyl is substituted with an hydroxy or a halogen on the 1 -position of the alkyl group.
  • R is 1 -hydroxyethyl, where the hydroxyl group is optionally protected.
  • R is a 1 -fluoroethyl.
  • R is 1 -hydroxyethyl.
  • R is an acylamino group, particularly an acylamino group of a cephalosporin, cephamycin, carbacephem or oxacephem antibiotic.
  • R is R1-CO-NH-, where Ri is any of various organic groups, including without limit, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted carbocyclic, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl.
  • R1CONH- is an acylamino group of a known beta-lactam antibiotic. A wide variety of beta-lactam antibiotics is known in the art.
  • Ri is an optionally substituted benzyl group.
  • the compounds of Formula I are cephalosporins, where M is Mi , Zi is S and R B is hydrogen.
  • the compounds of Formula I are cephalosporins, where M is Mi , Zi is S, R is an acylamino group and R B is hydrogen.
  • M is Mi, Zi is S, R B is hydrogen and both of R A are hydrogen.
  • M is Mi , Zi is S, R is an acylamino group, R B is hydrogen and both of R A are hydrogen.
  • both R A are hydrogen.
  • one R A is a C1 -C3 alkyl and the other R A is hydrogen.
  • one R A is a methyl and the other R A is hydrogen.
  • the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and R B is C1 -C3 alkyoxy.
  • the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and R B is C1 -C3 alkyoxy.
  • the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and R B is methoxy.
  • the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and R B is methoxy.
  • both R A are hydrogen.
  • one R A is a C1 -C3 alkyl and the other R A is hydrogen.
  • one R A is a methyl and the other R A is hydrogen.
  • the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CH2 or CH(CH3). In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2, or more specifically Z2 is CH2 or CH(CH3) and R B is hydrogen. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2, or more specifically Z2 is CH2 or CH(CH3), R B is hydrogen and R is a C1 -C6 hydroxyalkyl group.
  • the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CH2 or CH(CH3), R B is hydrogen and R is a 1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, R is an (R)-1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CH2 or CH(CH3), R B is hydrogen and R is a 1 -hydroxyethyl group. In specific embodiments, R is an (R)-1 -hydroxyethyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(R E )2 or more specifically Z2 is CH2 or CH(CH3), R B is hydrogen and R is an acylamino group.
  • the compounds of Formula I are monobactams where M is M3 and n is 0.
  • compounds of Formula I have a cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have a 5- or 6- member cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have an isoquinoline diol or a quinolone diol group.
  • -CFI2-X is:
  • R11 and R12- R-is are generally as defined in Formula Xi or
  • R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
  • R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
  • Ri6 is hydrogen or unsubstituted or substituted C1 -C8 alky or cycloalkyl;
  • Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl
  • Ri8 is a divalent -(CH2) P - moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and R18 together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds.
  • the phenyl ring at the right of the Formula X2 is substituted with at least two hydroxyl groups on adjacent ring carbons and is bonded to the positively charged N through the R18 moiety or is fused to the 5- or 6-member ring formed by R17 and R18.
  • optional substitution is substitution with one or more C1 -C3 alkyl group, C1 -C3 alkoxy group, hydroxyl, halogen, carboxylate or esters thereof, nitro, or cyano.
  • Specific halogens include chlorine or fluorine.
  • the M group of Formula I is selected from any Mi , M2, or M3 group set forth below:
  • the Mi group is selected from M1-1-M1-21 as set forth below:
  • the M2 group is selected from M2-1-M2-15 as set forth below:
  • the M3 group is selected from M3-1-M3-25 as set forth below:
  • R M is hydrogen or a C1 -C6 alkyl group, particularly an unsubstituted C1 -C6 alkyl group. In certain of these embodiments, R M is a C1 -C3 unsubstituted alkyl group and particularly a methyl group. In certain embodiments of M set forth above, each R D is hydrogen. In certain embodiment of M set forth above, R c is hydrogen or a pharmaceutically acceptable cation, when the CO2 group is negatively charged.
  • R c is unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, or acyloxy-substituted alkyl.
  • R c is hydrogen.
  • M is selected in the alternative from M1-2 to M1-10, M2-8 to M2-13, or M3-6 to M3-25.
  • M is selected in the alternative from M3-7, M3-9, M3-13 to M3-19.
  • M is selected in the alternative from M3-20 to M3-25.
  • M is selected from M3-8, M3-10, M3-11 , or M3-12. In certain embodiments, M is selected from M1-7, M1-9, M2-12, M2-13, M3-10, M3-12, M3-16, or lVh-is.
  • the B ring of Formula I is selected from any B ring set forth below:
  • A indicates optional substitution of one or more non-hydrogen substituents on the indicated ring or optional substitution of a single non-hydrogen substituent on a given ring position.
  • Optional non-hydrogen substituents A are selected from one or more nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, and C1-C3 trialkylammonium.
  • More preferred A are nitro, halide, methyl, methoxy, hydroxyl, methyl sulfonyl or amino.
  • Specific A groups are nitro, chloro, and bromo.
  • the B ring is other than a B ring having a positively charged nitrogen at the ring position attached to the M moiety or the -CH2-X moiety.
  • the B ring is selected from one of B1-B7, B8-B25, B26-B33, B34-B38, or B38-B58. In certain embodiments, the B ring is selected from one of Bi, B4, B7, Be, Bn, Bn, B17, B26, or B27-B38. In certain embodiments, the B ring is one of Bi, B4, or B7, where A is halogen or nitro, Be, Bn, Bn, or Bi7. In certain embodiments, the B ring is selected from Bi, B7, Be, and Bn. In certain embodiments, the B ring is selected from B39 or B40. In certain embodiments, the B ring is other than B39 or B40. In certain embodiments, the B ring contains one N ring atom. In certain embodiments, the B ring contains two N ring atoms.
  • the M group of Formula I is selected from any M group set forth above and the B ring of Formula I is selected from any B ring as set forth above.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi , B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, or Bi4.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, B 14 , B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the M group of Formula I is selected from M3-10, or M3-12, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the M group of Formula I is selected from M3-23, or M3-25, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36.
  • the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32.
  • the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, or B14.
  • the -CFI2X group of any compound of Formula I, II, III, IV, or V is selected from:
  • the Xs of any compound of Formula I, II, III, IV, or V is selected from:
  • the CQ of any compound of Formula I, II, III, IV, or V is selected from:
  • the X7 of any compound of Formula I, II, III, IV, or V is selected from:
  • the Xe of any compound of Formula I, II, III, IV, or V is selected from:
  • the X9 of any compound of Formula I, II, III, IV, or V is selected from:
  • the X10 of any compound of Formula I, II, III, IV, or V is selected from:
  • the X11 of any compound of Formula I, II, III, IV, or V is selected from:
  • the Xi2 of any compound of Formula I, II, III, IV, or V is selected from:
  • R12-R15 and Ri6 are as defined in Formula Xi or X2, and each Q is independently selected from N or C-Ax. Each W is independently selected from N-RN or C(AX)2.
  • AX represents optional substitution with one or two non-hydrogen substituents on carbon atoms in the indicated ring or optional substitution with one or two non-hydrogen substituents on a given carbon ring position.
  • Ax is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -C02-alkyl ester, -COFI, -COCH3, phenyl, and benzyl.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, and methoxy.
  • Ax represents no substitution.
  • Ax represent substitution on one ring carbon with OH, halogen (particularly Cl) or methyl.
  • R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate, carboxylate ester, or acyl groups. In specific embodiments, R12-R15 are all hydrogens. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol). In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12- Ri5 are hydrogen, hydroxyl, methyl, or halide. In specific embodiments, any two R12- Ri5 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, or halide. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen, chloride or both hydrogen or both chloride.
  • R16 is a C1 -C6 alkyl or cycloalkyl group. In specific embodiments, R16 is hydrogen. In specific embodiments, R16 is a methyl group.
  • the -CH2-X group is selected from: X5; CQ; XS-I ; CQ-I ; XS-2, X5-3, X5-4, X5-5, or XS-Q; CQ-2, CQ-3, CQ-4, CQ- 5, or CQ-Q; C7, Cb, CQ, CIO, Xu or C12; C7-1 ; Cb-i ; CQ-I , or C9-2; C10-1 or C10-2; C11-1 or C11-2; or C12-1 or Xi2-2, wherein in each case variables have values as defined above.
  • R12-R15 and R16 are as defined in Formula Xi or X2.
  • Ax represents optional substitution with one or more non-hydrogen substituents on carbon atoms in the indicated ring or a single optional non-hydrogen substituent on a given carbon ring position, and RN is hydrogen or a C1 - C3 alkyl group.
  • Ax is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -C02-alkyl ester, -COH, -COCH3, phenyl, and benzyl.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3.
  • optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, and methoxy.
  • Ax represents all hydrogens.
  • Ax represents a single non-hydrogen substituent which is selected from halide, nitro, hydroxyl, methyl or methoxy.
  • R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate, carboxylate ester, or acyl groups. In specific embodiments, R12-R15 are all hydrogens. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol). In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, hydroxyl, methyl, or halide.
  • any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, or halide. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen, chloride or both hydrogen or both chloride.
  • R16 is a C1 -
  • R16 is hydrogen. In specific embodiments, R16 is a methyl group.
  • RN is hydrogen. In other embodiments, RN is methyl.
  • the -CH2-X group is selected from those where the X group is a cation selected from X100, X120, X130, Xi4o, Xi5o, X160, Xi7o, or X180; X101-X109; X121-X129; X131-X139; X141-X149; X151-X159; X16I- X169; X171-X179; X181-X189; X107-X109; X127-X129; X137-X139; X147-X149; X157-X159; X167-X169; X177-X179; or X187-X189; wherein in each case variables have values as defined for the amine cations set forth above.
  • Ay represents optional substitution with one or more non-hydrogen substituents on carbon atoms in the indicated ring or a single optional non-hydrogen substituent on a given carbon ring position.
  • Ay is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1-C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl.
  • optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -CC -alkyl ester, -COH, -COCH3, phenyl, and benzyl.
  • optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3.
  • optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, and methoxy.
  • Ay represents all hydrogens.
  • Ay represents a single non-hydrogen substituent which is selected from halide, nitro, hydroxyl, methyl or methoxy.
  • RM represents hydrogen or an optionally substituted C1 -C6 alkyl. In certain embodiments, RM is an optionally substituted C1 -C3 alkyl group. In certain embodiments, RM is unsubstituted alkyl. In certain embodiments, RM is methyl. In certain embodiments, RM is hydrogen.
  • RN represents optionally substituted C1 -C6 alkyl groups or two or three of RN together with the nitrogen to which the RN are bound forms a 4-10 member optionally substituted heterocyclic ring.
  • RN is unsubstituted C1 -C6 alkyl.
  • Rx is selected from hydrogen, optionally substituted C1 -C6 alkyl, and optionally substituted C1 -C6 acyl groups, where optional substitution includes substitution with one or more hydroxyl, amino, alkyl amino, or dialkyl amino groups.
  • Rx are alkyl amino- or dialkyl amino-substituted C1 - C6 acyl groups.
  • monobactams of Formula I have -CH2-X as set forth above, and in certain of these embodiments n is 1 and L is ethylene.
  • -CH2- X is selected from: X200-X213; X200-X205; X206; X207; X208; X209; X210; X211, X212; or X213, wherein in each case variables have values as defined above.
  • Ri groups set forth above include exemplary precursor carboxylic acids R1-COOH that are typically employed for reaction with corresponding amines (see Synthesis Schemes 1 -5 and the Examples below) to introduce the R1-CO- moiety into compounds of the disclosure. Methods for such introduction are known in the art and are exemplified herein.
  • Ri is a group selected from those of compounds R1-4 - R1-22.
  • Ri is a group selected from those of compounds R1-4 - R1-16
  • Ri is a group selected from those of compounds R1-5, Ri- 6, R1-7, R1-8, R1-9, R1-10, R1-11 , or R1-12.
  • Ri is a group selected from those of compounds R1-11 or R1-12. In certain embodiments, Ri is a group selected from those of compounds R1-13 or R1-14. In certain embodiments, Ri is a group selected from those of compounds R1-15 or R1-16. In certain embodiments, Ri is a group selected from those of compounds R1-17 or R1-18. In certain embodiments, Ri is a group selected from those of compounds R1-19 or R1-22. In certain embodiments, Ri is a group selected from those of compounds R1-24 or R1-25. In certain embodiments, Ri is a group selected from those of compounds R1-23, or R1-26 - R1-47. In certain embodiments, Ri is a group selected from those of compounds R1-38 - R1-43.
  • the compounds provided herein have a structure set forth below:
  • R-i , R c , X are as defined for Formula I.
  • X is a halogen, and in other embodiments X is other than a halogen.
  • R c is hydrogen or -CO-OR c is carboxylate.
  • R c is and optionally substituted phenyl group.
  • the compounds provided herein are any one of Formulas I-60 to 1-123 set forth above.
  • the disclosure provides antibacterial compounds of Formulas I-60 to I-72; I-73 to I-75; I-76 to I-78; I-79 to 1-81 ;
  • the compounds set forth above can be illustrated as positively charged species (and X groups listed can be positively charged species) where no specific anion is shown. It is appreciated in the art that such compounds are prepared as salts, for example, where the anion of the salt is a pharmaceutically acceptable cation.
  • Certain compounds provided herein carry a -CO-0-R c group which can be a carboxylate group. In certain embodiments, these compounds are zwitterionic.
  • the compounds of these embodiments may be in zwitterionic form or may be in the form of an ammonium cation with an appropriate anion, such as a halide (e.g. , Cl-, Br or h), an organic anion, such as sulfate, bisulfate, acetate or trifluoroacetate, or any pharmaceutically acceptable anion, such as described herein.
  • R 1 is of formula:
  • R22 is as defined for Ri in Formula I
  • X is N or CRx, where Rx is hydrogen, C1 - C3 alkyl or halogen, and particularly chlorine
  • Re is hydrogen or (Rp)2PO-, where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both Rp are methyl.
  • the R 1 group is in the E or Z conformation with respect to the oxime group. More specifically the R1-1 group is in the Z conformation.
  • R 1 is of formula:
  • Ra and Rb are as defined for Ri in Formula I
  • X is N or CRx, where Rx is hydrogen, C1 -C3 alkyl or halogen, and particularly chlorine, and Re is hydrogen or (Rp)2PO- where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both are methyl. More specifically, Ra and Rb are hydrogen or methyl and Re is hydrogen. Yet more specifically, Ra and Rb are both hydrogen or both methyl groups and Re is hydrogen.
  • X is CH or N.
  • R 1 is of formula:
  • Rx is hydrogen, C1 -C3 alkyl or halogen, and particularly chlorine
  • Re is hydrogen or (Rp)2PO-, where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both are methyl.
  • Rf is hydrogen, hydroxyl, C1 -C3 alkyl, C1 -C3 alkoxy, unsubstituted or substituted phenyl or unsubstituted or substituted benzyl.
  • Rg, Rh and Ri are independently selected from hydrogen, hydroxyl, amino, alkyl amino. More specifically, one or two of Rg, Rh, Ri are hydroxyl and the others are hydrogen. More specifically, Re is hydrogen. More specifically, Rf is hydroxyl or alkoxy. More specifically, Rf is hydroxyl.
  • Ri and R c are as defined for Formula I;
  • each Q is independently, N or CH; and -CFI-X2 is
  • Rii and R12- R-is are generally as defined in Formula Xi or
  • R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
  • R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
  • R16 is hydrogen or unsubstituted or substituted C1 -C6 alkyl
  • Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl
  • R18 is a divalent -(CH2) P - moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and Rie together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds.
  • the X group is any one of the cationic groups of Xioo-Xi89.
  • the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47.
  • the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1, where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • the compounds provided herein have the structure set forth in Formula 11-1 :
  • Ri and R c are as defined for Formula I; and each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors provided herein. In embodiments of Formula 11-1 , the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above. In embodiments of Formula 11-1 , the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula 11-1 , the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula 11-1 , both Q are CH. In embodiments of Formula 11-1 , one Q is N and the other is CH. In embodiments of Formula 11-1 , both Q are N.
  • Ri is R1-11 , R c is hydrogen, and Q is CH.
  • R1 is R1-5, R c is hydrogen, and one Q is CH and the other Q is N.
  • the compounds provided herein have the structure set forth in Formula II-2:
  • Ri and R c are as defined for Formula I; and each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors provided herein. In embodiments of Formula II-2, the Ri group is any one of Ri-i , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula II-2, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula II-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula II-2, both Q are CH. In embodiments of Formula II-2, one Q is N and the other is CH. In embodiments of Formula II-2, both Q are N.
  • the compounds provided herein have the structure set forth in Formula II-3:
  • Ri and R c are as defined for Formula I;
  • each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors provided herein.
  • the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1, where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • Ri is R1-11
  • R c is hydrogen
  • Q is CH.
  • R1 is R1-5, R c is hydrogen, and Q is CH.
  • the compounds provided herein have the structure set forth in Formula III:
  • Ri is as defined for Formula I;
  • X2 is as defined for Formula II.
  • each Q is independently, N or CH.
  • the X group is any one of the cationic groups of X100-X189.
  • the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47.
  • the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • the -CFI2-X group is any one of the -CFI2-X groups of X200-X213.
  • the compounds provided herein have the structure set forth in Formula III-1
  • Ri is as defined for Formula I and
  • each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • the compounds provided herein have the structure set forth in Formula MI-2:
  • Ri is as defined for Formula I and
  • each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47.
  • the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1, where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • the compounds provided herein have the structure set forth in Formula MI-3:
  • Ri is as defined for Formula I;
  • each Q is independently, N or CH.
  • the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • Ri and R c are as defined for Formula I;
  • X2 is as defined in Formula II; each Q is independently, N or CH; and the double bond is either cis or trans.
  • the X group is any one of the cationic groups of X100-X189.
  • the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R-M , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • the -CFI2-X group is any one of the -CFI2-X groups of X200-X213.
  • the compounds provided herein have the structure set forth in Formula IV-1 :
  • Ri and R c are as defined for Formula I; each Q is independently, N or CH; and the double bond is either cis or trans.
  • the Ri group is derived from any one of the carboxylic acid precursors of R-M - R1-47. In embodiments of Formula IV-1 , the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV-1 , the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula IV-1 , the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV-1 , both Q are CH. In embodiments of Formula IV-1 , one Q is N and the other is CH. In embodiments of Formula IV-1 , both Q are N.
  • the compounds provided herein have the structure set forth in Formula IV-2:
  • Ri and R c are as defined for Formula I;
  • each Q is independently, N or CH; and the double bond is either cis or trans.
  • the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47. In embodiments of Formula IV-2, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV-2, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula IV-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV-2, both Q are CH. In embodiments of Formula IV-2, one Q is N and the other is CH. In embodiments of Formula IV-2, both Q are N.
  • the compounds provided herein have the structure set forth in Formula IV-3:
  • Ri and R c are as defined for Formula I; each Q is independently, N or CH; and the double bond is either cis or trans.
  • the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • R and R c are as defined for Formula I;
  • X2 is as defined in Formula II;
  • each Q is independently, N or CH.
  • the X group is any one of the cationic groups of X100-X189.
  • the R is R1 CON H- and the R i group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • the -CH2-X group is any one of the -CH2-X groups of X200-X213.
  • the compounds provided herein have the structure set forth in Formula V-1 :
  • R and R c are as defined for Formula I;
  • each Q is independently, N or CH.
  • the R is R1 CONFI- and R i group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of Ri- 2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • the compounds provided herein have the structure set forth in Formula V-2:
  • R and R c are as defined for Formula I;
  • X2 is as defined in Formula II;
  • each Q is independently, N or CH.
  • R is R1CONFI- and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1 -1 , R1 -2 or R1 -3, where variables are as defined above.
  • the Ri group is any one of R1-1, where variables are as defined above.
  • the Ri group is any one of R1-2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • the compounds provided herein have the structure set forth in Formula V-3:
  • R and R c are as defined for Formula I;
  • X2 is as defined in Formula II;
  • each Q is independently, N or CH.
  • R is R1CONH- and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47.
  • the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above.
  • the Ri group is any one of R1-1 , where variables are as defined above.
  • the Ri group is any one of Ri- 2, where variables are as defined above.
  • both Q are CH.
  • one Q is N and the other is CH.
  • both Q are N.
  • R5, R6, R9, and Re are hydrogen
  • R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen and Ax is hydrogen;
  • R c is hydrogen
  • R is Ri, wherein Ri is
  • Ri is selected from the group consisting of:
  • R1-1 where R22 is a C1-C6 alkyl group
  • R1-1 where R22 is methyl
  • R1-1 where X is CH
  • R1-1 where X is CH and R22 is a 1-6 carbon alkyl group
  • R1-1 where X is N and R22 is a 1-6 carbon alkyl group
  • R1-1 where X is CH and R22 is methyl
  • R1-1 where X is N and R22 is methyl
  • R1-1 where X is CH, R22 is methyl and R1-1 is in the Z configuration;
  • R1-1 where X is N, R22 is methyl and R1-1 is in the Z configuration;
  • Ra and Rb are both C1 -C3 alkyl groups
  • R1-2 where X is CH and Ra and Rb are both methyl groups
  • R1-2 where X is N and Ra and Rb are both methyl groups; and benzyl.
  • R is selected from the group consisting of:
  • R1CO-NH- where Ri is R1-1 , X is CH and R22 is a methyl group;
  • R1CO-NH- where Ri is R1-1 , X is CH, R22 is a methyl group and the R1-1 is in the Z conformation;
  • R1CO-NH- where Ri is R1-1 , X is N and R22 is a methyl group;
  • R1CO-NH- where Ri is R1-1 , X is N, R22 is a methyl group and the R1-1 is in the Z conformation;
  • R1CO-NH- where Ri is R1-2 and X is CH and Ra and Rb are both methyl groups; R-iCO-NH-, where Ri is R1-2 and X is N and Ra and Rb are both methyl groups; a C1 -C6 alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group; a C1 -C6 alkyl group substituted with a hydroxy group or a protected hydroxyl group; a C1 -C6 alkyl group substituted with a hydroxy group; a 1 -hydroxyethyl group; and
  • both Q are CH; one Q is CH and the other is N; or both Q are N.
  • R c is selected from the group consisting of: hydrogen;
  • exemplary compounds of the disclosure which exhibit antibacterial activity as described herein are selected from Compound 504, 501 ,
  • the compounds are illustrated as positively charged species and it is appreciated in the art that such compounds are prepared as salts, for example, where the anion of the salt is a pharmaceutically acceptable cation.
  • Exemplary compounds provided herein include:
  • the compounds disclosed herein are useful for the synthesis of other compounds of Formula I. More specifically, compounds herein wherein X is Cl, Br, I, or an activated ester and wherein other potentially labile groups are protected with appropriate protecting groups can be used in syntheses described herein to prepare other compounds of Formula I.
  • Certain compounds of the disclosure may contain one or more stereogenic centers resulting in possible stereoisomers. It will be understood that for any such compound of the disclosure that all individual stereoisomer, any diastereomers and all mixtures thereof are included in the compounds as claimed. For any such compound the compound may be a racemic mixture of stereoisomers or may be a mixture which contains an excess of one stereoisomer and is thus optically active.
  • E and Z isomers of compounds of the disclosure may contain approximately equal amounts of E and Z isomers or may contain an excess of the E or Z an excess of the Z isomer.
  • the amounts of E and Z isomers in a given mixture can be readily determined by standard analytical techniques, such as NMR (nuclear magnetic resonance) methods.
  • a compound that is a given isomer (E or Z) that is free of the other isomer contains less than 2% by weight of the other isomer.
  • a compound that is given isomer (E or Z) that is free of the other isomer preferably contains less than 1 % by weight of the other isomer.
  • a compound that is a given isomer (E or Z) that is free of the other isomer more preferably contains less than 0.5% by weight of the other isomer. It will be appreciated in the art that when it is desired to employ a compound or intermediate that is a given E or Z isomer that it can be also desirable to minimize the amount of the other isomer present. It will however also be understood that small, albeit detectible levels of the other isomer that may be present may not be detrimental to the utility of the desired isomer.
  • two variables substituted on adjacent carbons in an alkyl group or on a ring can optionally together with the atoms (typically carbons or nitrogens) to which they are bonded form a carbocyclic or heterocyclic ring.
  • the ring can be a 5-member ring or a 6-member ring and may contain 0, 1 2 or 3 heteroatoms.
  • Preferred heteroatoms for such rings are nitrogen, oxygen and sulfur.
  • Such rings are optionally substituted, for example, with one or more halogens, hydroxyl, C1 -C3 alkyl, or carboxylate or esters thereof, or contain an oxo group (-CO-) in the ring.
  • such rings are unsubstituted.
  • Such rings may be unsaturated (i.e., contain no double or triple bonds), partially unsaturated, i.e. , contain one or two double bonds or may be aromatic (as understood in the art).
  • Groups herein are optionally substituted. Most generally alky, alkenyl, and aryl, heteroaryl, and heterocyclyl groups are optionally substituted, for example, with one or more oxo group, thioxo group, halogen, nitro, cyano, cyanate, azido, thiocyano, isocyano, isothiocyano, sulfhydryl, hydroxyl, alkyl, alkoxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, aryl, aryloxy, amino (-NH2), heteroaryl, heteroaryloxy, carbocyclyl, carbocyclyloxy, heterocyclyl, heterocyclyloxy, alkylthio, alkenylthio, alkynylthio, arylthio, thioheteroaryl, thioheteroaryl, thiocarbocyclyl, thiohetero
  • Organic groups of non hydrogen substituents are in turn optionally substituted with one or more halogens, nitro, cyano, isocyano, isothiocyano, hydroxyl, sulfhydryl, haloalkyl, hydroxyalkyl, amino, alkylamino, dialkylamino, arylalkyl, unsubstituted alkyl, unsubstituted alkenyl, unsubstituted alkynyl alkylalkenyl, alkylalkynyl, haloaryl, hydroxylaryl, alkylaryl, unsubstituted aryl, unsubstituted carbocyclic, halo-substituted carbocyclic, hydroxyl- substituted carbocyclic, alkyl-substituted carbocyclic, unsubstituted heterocyclic, unsubstituted heteroaryl, alkyl-substituted heteroaryl, or alkyl substituted hetero
  • organic groups of non-hydrogen substituents are not further substituted.
  • R' groups of substituents are independently selected from alkyl groups, haloalkyl groups, phenyl groups, benzyl groups and halo- substituted phenyl and benzyl groups.
  • non-hydrogen substituents have 1 -10 carbon atoms, 1 -7 carbon atoms, 1 -5 carbon atoms or 1 -3 carbon atoms.
  • non-hydrogen substituents have 1 -10 heteroatoms, 1 -6 heteroatoms, 1 -4 heteroatoms, or 1 , 2, or 3 heteroatoms. Heteroatoms preferably are O, N or S.
  • substituent groups include heterocyclic groups having a single 5- or 6-member ring containing 1 , 2or 3 heteroatoms selected from N, O or S and in which one or more ring carbons or nitrogens are optionally substituted with a C1 -C3 alkyl group.
  • optional substitution is substitution with 1 , 2, 3, 4 or 5 non-hydrogen substituents. In specific embodiments, optional substitution is substitution with 1 or 2 non hydrogen substituents. In specific embodiments, optional substitution is substitution with 1 non hydrogen substituents. In specific embodiments, optional substitution is substitution by one or more halogen, hydroxyl group, cyano group, nitro group, oxo group, thioxo group, unsubstituted C1 -C6 alkyl group or unsubstituted aryl group.
  • optional substitution of phenyl rings includes substitution with one or more halogen, one or more hydroxyl, one or more C1 -C3 alkyl, one or more C1 -C3 alkoxy, one or more nitro, or one or more cyano.
  • optional substitution of phenyl rings includes substitution with one or more halogen, one or more hydroxyl, one or more C1 -C3 alkyl, or one or more C1 -C3 alkoxy.
  • optional substitution of phenyl rings includes substitution with one or more halogen, one or more hydroxyl, or one or more C1 -C3 alkyl.
  • optional substitution of phenyl rings includes substitution with one halogen, one hydroxyl, one C1 -C3 alkyl, one C1 -C3 alkoxy, one nitro, and/or one cyano.
  • any of the above groups which contain one or more substituents it is understood, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
  • the compounds of this disclosure include all stereochemical isomers arising from the substitution of these compounds.
  • Compounds of the disclosure may contain chemical groups (acidic or basic groups) that can be in the form of salts.
  • Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, for example, trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides (formed with hydrogen bromide), hydroiodides, 2-hydroxyethanesulfonates,
  • Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as benzathines, dicyclohexylamines, hydrabamines [formed with N,N-bis(dehydro- abietyl)ethylenediamine], N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, and salts with amino acids such as arginine, lysine and the like.
  • organic bases for example, organic amines
  • organic amines such as benzathines, dicyclohexylamines, hydrabamines [formed with N,N-bis(dehydro- abietyl)ethylenediamine], N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, and salts with amino acids
  • Basic nitrogen- containing groups may be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.
  • lower alkyl halides e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and iodides
  • dialkyl sulfates e.g., dimethyl, diethyl, dibutyl, and diamyl sulf
  • Compounds of the disclosure may be in the zwitterionic form.
  • Salts of the disclosure include “pharmaceutically acceptable salts” which refers to those salts which retain the biological effectiveness and properties of the free bases or free acids, and which are not biologically or otherwise undesirable.
  • Pharmaceutically acceptable salts comprise pharmaceutically-acceptable anions and/or cations.
  • compositions of the disclosure can be administered in the form of pharmaceutically acceptable salts which include the following non-limiting examples: alkali metal salts, such as those of lithium, potassium and sodium; alkali earth metal salts, such as those of barium, calcium and magnesium; transition metal salts, such as those of zinc; and other metal salts, such as those of aluminum, sodium hydrogen phosphate and disodium phosphate; salts of nitrates, borates, methane sulfonates, benzene sulfonates, toluene sulfonates, salts of mineral acids, such as those of hydrochlorides, hydrobromides, hydroiodides and sulfates; salts of organic acids, such as those of acetates, trifuoroacetates, maleates, oxalates, lactates, malates, tartrates, citrates, benzoates, salicylates, ascorbates, succinates, butyrates, valerates
  • compound of any formula herein are salts wherein the anion of the salt is selected from halide, more specifically chloride or bromide, acetate, tosylate, tartrate, sulfate, bisulfate, succinate, phosphate, oxalate, nitrate, mesylate, maleate, malate, and citrate.
  • compounds of any formula herein are salts of chloride, bromide, sulfate, bisulfate, or acetate.
  • compounds of the disclosure are hydrates of the compounds herein and more specifically are hydrates of salts of the compounds herein.
  • Pharmaceutically acceptable salts of the compounds of the disclosure can be derived from inorganic or organic acids or can be derived from inorganic or organic bases as is known in the art.
  • Basic amino acids useful for salt formation include arginine, lysine and ornithine.
  • Acidic amino acids useful for salt formation include aspartic acid and glutamic acid.
  • esters which include, among others, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids and boronic acids.
  • solvates can form complexes with solvents in which they are reacted or from which they are precipitated or crystalized. These complexes are known as "solvates".
  • solvates For example, a complex with water is known as a "hydrate”.
  • Solvates, and more particularly hydrates, of the cephem compounds of the disclosure are within the scope of the disclosure.
  • Pharmaceutically acceptable solvates and hydrates are complexes of a compound of the disclosure with one or more solvent or water molecules, or 1 to about 100, or 1 to about 10, or one to about 2, 3 or 4, solvent or water molecules.
  • Certain groups in the compounds of the disclosure are optionally protected, for example, hydroxyl, carboxylate and amine groups. Methods of protection of such groups is known in the art and employed generally during synthesis again as is known in the art. T.W. Green et al. (1999) Protecting Groups in Organic Synthesis (Wiley Interscience) provides an overview of the types of protecting groups and their use. This reference is incorporated by reference herein for descriptions of types of protecting groups and methods for protecting and deprotecting.
  • compounds of the disclosure may contain a protected hydroxyl group or a protected carboxyl group.
  • Exemplary hydroxyl protecting groups include alkoxyalkyl groups (e.g., methoxym ethyl, forming an ether at the hydroxy), C1 -C3 alkyl or C2-C6 alkenyl or arylalkyl or silyl groups (again forming ethers), or -CO-alkyl or -CO-arylalkyl (forming esters.
  • alkoxyalkyl groups e.g., methoxym ethyl, forming an ether at the hydroxy
  • C1 -C3 alkyl or C2-C6 alkenyl or arylalkyl or silyl groups (again forming ethers)
  • -CO-alkyl or -CO-arylalkyl forming esters.
  • hydroxyl protecting groups include, among others, -COCFI3, -CO-t-butyl, -CO-phenyl (optionally substituted at the phenyl ring), -CO-benzyl (optionally substituted at the phenyl ring), optionally substituted benzyl, allyl, silyl (trialkylsilyl, diarylalkysilyl, aryldialkylsilyl, e.g., trimethylsily, diphenylmethylsilyl, phenyldimethylsilyl, and the like). Hydroxy groups may also be protected by formation of acetals. Carboxylate groups may be protected as esters.
  • the compounds provided herein are in their tautomeric forms.
  • the compounds of Formula I are one of two tautomeric forms.
  • the tautomeric form of the compound influences the elimination rate and/or half-life of the compound.
  • one tautomeric form can have decreased elimination rates and/or increased drug half- life as compared to the other tautomeric form.
  • methods are provided for treating a bacterial infection in a subject by administering one or more compounds or pharmaceutical compositions provided herein.
  • the subject is a human or non-human animal, in particular a mammal.
  • GL-332, GL-337, GL-357, or GL-352 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof is administered to a subject to treat a bacterial infection.
  • the subject is administered GL-332 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof.
  • the subject is administered GL-357 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof.
  • the subject is administered GL-337 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof.
  • the subject is administered GL-352 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof.
  • the disclosure relates, in particular, to cephem and penem compounds which exhibit strong antibacterial activity against certain multi-drug resistant bacteria.
  • compounds herein exhibit low pgram/mL Minimum Inhibitory Concentrations (MIC) against bacteria which produce extended spectrum beta- lactamases.
  • compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one cephalosporin antibiotic.
  • compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one cephem or penem antibiotic.
  • compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one carbapenem.
  • compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which produce at least one ESBL.
  • compounds of the disclosure exhibit antibiotic activity against Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-positive bacteria. In specific embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which produce various beta-lactamases. In embodiments, compounds of the disclosure, in particular, exhibit antibiotic activity against beta-lactamase producing Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against a wide range of Pseudomonas aeruginosa strains, including clinical isolates.
  • compounds of the disclosure exhibit antibiotic activity against Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-positive bacteria. In specific embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which produce various beta-lactamases. In embodiments, compounds of the disclosure in particular exhibit antibiotic activity against beta-lactamase producing Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against a wide range of Pseudomonas aeruginosa strains, including clinical isolates.
  • compounds of the disclosure exhibit antibiotic activity against Enterobacteriaceae, including but not limited to strains of Escherichia coli ; Klebsiella ; Proteus, Citrobacter, Serratia ; and/or Enterobacter.
  • the compounds provided herein exhibit antibiotic activity against strains of Klebsiella pneumoniae ; Klebsiella oxytoca ; Proteus mirabilis ; Citrobacter freundii ; Serratia marcescens ; Enterobacter aerogenes ; and/or Enterobacter cloacae.
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL).
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce ESBL selected from TEM, SHV, and CTX-M.
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce ESBL selected from TEM-26, SHV-1 , CTX-M-14, and CTX-M-15.
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases [24]
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases selected from DHA enzymes, particularly DHA-1 .
  • compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemases. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemases selected from KPC, VIM, IMP, NDM, and OXA. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemase selected from KPC- 2, KPC-3, VIM-1 , IMP-1 , NDM-1 , OXA-48 and OXA-58.
  • the disclosure provides methods for treating or preventing bacterial infection and the symptoms and disorders associated therewith.
  • the disclosure provides pharmaceutical compositions comprising a pharmaceutically effective amount of one or more compounds and/or salts and/or solvates of the disclosure and a pharmaceutically acceptable carrier or excipient.
  • the compounds of the disclosure are of particular use in the treatment of infections of bacteria which exhibit resistance to or more beta-lactam antibiotics, particularly those that exhibit resistance to one or more cephalosporins and particularly those that exhibit resistance to a carbapenem.
  • Compounds of the disclosure are of particular use in the treatment of infections of bacteria which generate one or more ESBL.
  • Compounds of the disclosure exhibit resistance to one or more ESBL.
  • Compounds of the disclosure are of particular use in the treatment of infections of bacteria which generate one or more ESBL.
  • Compounds of the disclosure are of particular use in the treatment of infections of Gram-negative bacteria.
  • Compounds of the disclosure are of particular use in the treatment of infections of Pseudomonas strains and particularly strains of Pseudomonas aeruginosa.
  • the compounds, salts and solvates thereof of the disclosure can be used to prepare medicaments for the treatment and prevention of bacterial infection and the symptoms and disorders associated therewith.
  • the compounds, salts and solvates thereof of the disclosure can be used in particular to prepare medicaments for the treatment and prevention of bacterial infection of bacteria which produce one or more ESBL and the symptoms and disorders associated therewith.
  • the antibacterial compounds of this disclosure may be co-administered in combination with one or more antibacterial compounds, including antibacterial compounds which are cephems or penems other than those of Formulas l-lll herein and including antibacterial compounds which are not beta-lactam antibiotics.
  • the antibacterial compounds of this disclosure may be co-administered in combination with one or more beta-lactamase inhibitors other than the compounds of Formula I herein.
  • Co-administration includes among others separate administration of active ingredients at about the same time, combined administration in the same formulation, as well as sequential separate administration.
  • Compounds of the disclosure may be co-administered with a beta- lactamase inhibitor.
  • compounds of the disclosure may be co formulated with a beta-lactamase inhibitor.
  • a number of such inhibitors are known in the art and one of ordinary skill in the art understands how to co-formulate such inhibitors with a given beta-lactam antibiotic.
  • Beta-Lactamase inhibitors include among others avibactam, tazobactam, sulbactam, clavulanic acid, and relebactam.
  • Compounds of the disclosure may be co-administered with a monobactam, such as aztreonam.
  • Compounds of the disclosure may be co-administered with a carbapenem, other than a compound of Formula I, such as meropenem.
  • Compounds of the disclosure may be co-administered with a non-beta lactam antibiotic, such as an aminoglycoside antibiotic.
  • Compounds of the disclosure may be co-administered with an aminoglycoside antibiotic, such as amikacin, gentamicin, kanamycin, neomycin, or tobramycin.
  • the disclosure also relates to the use of one or more compound, salt or solvate of Formula I and any sub formula thereof for the treatment of a bacterial infection or a complication of a bacterial infection.
  • the disclosure further relates to the use of one or more pharmaceutical compositions comprising a compound, salt or solvate of Formula I and any sub formula thereof for the treatment of a bacterial infection or a complication of a bacterial infection.
  • the disclosure also relates to the use of one or more compound, salt or solvate of Formula I and any subformula thereof for the preparation of a medicament for the treatment of a bacterial infection or a complication thereof.
  • the disclosure further relates to the use of one or more pharmaceutical compositions comprising a compound, salt or solvate of Formula I and any sub formula thereof for the preparation of a medicament for the treatment of a bacterial infection or a complication thereof.
  • compositions comprising one or more of the compounds provided herein.
  • Pharmaceutical compositions herein comprise a therapeutically effective amount of a cephem compound of this disclosure.
  • compositions provided herein contain one or more of GL-332, GL-337, GL-357, or GL-352 stereoisomer, pharmaceutical acceptable salt, or tautomer.
  • compositions herein comprise a therapeutically effective amount of a cephem or penem compound of any chemical formula herein.
  • pharmaceutical compositions herein comprise a therapeutically effective amount of a cephalosporin of any chemical formula herein.
  • pharmaceutical compositions herein comprise a therapeutically effective amount of a cephalomycin of any chemical formula herein.
  • pharmaceutical compositions herein comprise a therapeutically effective amount of a carbapenem of any chemical formula herein.
  • pharmaceutical compositions herein comprise a combined therapeutically effective amount of two or more cephem or penem compounds of this disclosure.
  • pharmaceutical compositions herein comprise a combined therapeutically effective amount of two or more cephem or penem compounds of any formula herein and in particular any compound of Formula I.
  • compositions herein optionally contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitors, other than a compound of Formula I.
  • pharmaceutical compositions herein optionally contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitors, other than a cephem.
  • pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more beta-lactamase antibiotics other than a compound of Formula I.
  • pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more monobactam.
  • pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more carbapenem other than a compound of Formula I.
  • pharmaceutical compositions herein contain other appropriate active ingredients including, for example, aminoglycoside antibiotics.
  • Pharmaceutical compositions herein optionally further comprise one or more pharmaceutically acceptable carriers or excipients as are known in the art.
  • compositions or dosage forms can be administered by any known route that is appropriate for the patient being treated and for the treatment or prophylaxis that is desired.
  • administration can be orally or non-orally in the form of, for example, granules, powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixir, suspensions or solutions, by mixing these effective components, individually or simultaneously, with pharmaceutically acceptable carriers, excipients, binders, diluents or the like.
  • a solid formulation for oral administration can comprise one or more of the compounds or salts, or solvates thereof, of the disclosure alone or in appropriate combination with other active ingredients.
  • Solid formulations can be in the form of powders, granules, tablets, pills and capsules.
  • the instant compounds can be mixed with at least one additive, for example, sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi synthetic polymers or glycerides.
  • formulations can contain, as in conventional cases, further additives, for example, an inactive diluent, a lubricant such as magnesium stearate, a preservative such as paraben or sorbic acid, an anti-oxidant such as ascorbic acid, tocopherol or cysteine, a disintegrator, a binder, a thickening agent, a buffer, a sweetener, flavoring agent and/or a perfuming agent. Tablets and pills can also be prepared with enteric coating. Standard methods of formulation can be applied to preparation of formulations of the compounds and salts of the disclosure.
  • Non-oral administration includes subcutaneous injection, intravenous injection, intramuscular injections, intraperitoneal injection or instillation.
  • injectable preparations for example, sterile injectable aqueous suspensions or oil suspensions can be prepared by known methods.
  • compositions may be formulated as known in the art for administration by inhalation, such as in the form of a nasal aerosol or nasal spray, a dry powder spray or as a solution or suspension for inhalation and may be prepared as solutions or suspensions in saline or other suitable carrier, and benzyl alcohol or other suitable preservatives, absorption promoters, fluorocarbons, or solubilizing or dispersing agents.
  • Rectal suppositories can be prepared by mixing the drug with a suitable vehicle, for example, cocoa butter and polyethylene glycol, which is in the solid state at ordinary temperatures, in the liquid state at temperatures in intestinal tubes and melts to release the drug.
  • a suitable vehicle for example, cocoa butter and polyethylene glycol
  • liquid preparations for oral administration include pharmaceutically acceptable emulsions, syrups, elixirs, suspensions and solutions, which may contain an inactive diluent, for example, pharmaceutically acceptable water.
  • the pharmaceutical composition herein can be formulated for topical administration with a suitable ointment containing one or more of the compounds or salts of the disclosure suspended or dissolved in a carrier, which include mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and pharmaceutically acceptable water.
  • topical formulations can be formulated with a lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and pharmaceutically acceptable water.
  • dosages of therapeutic compounds are dependent on age, body weight, general health conditions, sex, diet, dose interval, administration routes, excretion rate, combinations of drugs and conditions of the diseases treated. While taking these and other necessary factors into consideration, generally, dosage levels of between about 10 pg per day to about 5000 mg per day, preferably between about 100 mg per day to about 1000 mg per day of the compound are useful in the treatment of bacterial infection.
  • the pharmaceutical compositions of this disclosure will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy.
  • the amount of active ingredient that may be combined with the carrier or excipient materials to produce a single dosage form will vary depending upon the patient/individual treated and the particular mode of administration.
  • a typical preparation will contain from about 5% to about 95% active compound (W/W).
  • such preparations contain from about 20% to about 80% active compound.
  • a desirable dosage form for antibiotic compounds of the disclosure is an oral dosage form to be administered once a day or twice a day.
  • Another desirable dosage form for antibiotic compounds of the disclosure is an parenteral dosage form to be administered once a day or twice a day.
  • any of the exemplary amine salts of Scheme 4 can be employed as starting materials in the synthetic methods herein.
  • Appendix A also provides additional methods for synthesis of compounds of Formula I herein where the R group is an acylamino group.
  • Scheme 6 herein provides exemplary carboxylic acid precursors for incorporation of various useful acyl amino groups into compounds of the disclosure wherein R is an acylamino group.
  • any of the exemplary carboxylic acids of Scheme 6 can be employed as starting materials in the synthetic methods herein.
  • Synthetic methods herein are specifically illustrated for the preparation of cephalosporins, monobactams and carbapenems. These exemplary methods can be readily adapted to the preparation of cephamycins, carbacephems, oxacephems, penems, and oxapenems.
  • Y and Z are independently CH or N.
  • pharmaceutically effective amount refers to an amount effective in treating a bacterial infection, or a symptom or complication thereof, in a patient (human or other mammal) either by administration of a single compound of Formulas I- III, or a salt or solvate thereof or in combination with other agents.
  • the pharmaceutically effective amount of a given compound when administered as the only active ingredient may differ from its pharmaceutically effective amount when administered with other active ingredients. It will be appreciated that the pharmaceutically effective amount of a compound may differ from that of a salt of the same compound.
  • treating includes the alleviation of symptoms of a particular disorder in a patient or a measurable improvement of a parameter associated with a particular disorder. Treating includes treatment to prevent a bacterial infection and to delay the progress of an infection.
  • prophylactically effective amount refers to an amount of a compound or salt of the disclosure effective in preventing a bacterial infection in a patient.
  • the compounds of the present disclosure are useful in the treatment of individuals infected by various bacteria (which include infections of combinations of different bacteria) for the prophylaxis of these individuals. It will be appreciated in the art, that individuals at risk for bacterial infection can be treated employing one or more of the compounds or salts of the disclosure to prevent or delay infection.
  • patient refers to any animal and more specifically to a mammal, including a human.
  • patient or subject is a human.
  • alkyl or“alkyl group,” alone or in combination, refer to a monoradical of a straight chain or branched saturated hydrocarbon.
  • Alkyl groups include straight-chain and branched alkyl groups. Unless otherwise indicated alkyl groups have 1 -12 carbon atoms (C1 -C12 alkyl groups) and preferred are those that contain 1 -6 carbon atoms (C1 -C6 alkyl groups) and more preferred are those that contain 1 -4 carbon atoms (C1 -C4 alkyl groups) and those that contain 1 -3 carbon atoms (C1 -C3 alkyl groups). Unless otherwise indicated alkyl groups are optionally substituted with one or more non-hydrogen substituents as described herein.
  • any alkyl group designated herein can be unsubstituted.
  • the designation of an alkyl group having a range of carbon atoms includes all isomers having that number of carbon atoms.
  • Exemplary alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, s-butyl, t- butyl, n-pentyl, branched pentyl, n-hexyl, branched hexyl, all of which are optionally substituted.
  • Substituted alkyl groups include fully halogenated or semihalogenated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms. Substituted alkyl groups include fully fluorinated or semifluorinated alkyl.
  • alkoxy refers to an -O-alkyl group, where alkyl is as defined above.
  • Alkoxy groups include among others methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert butoxy. Alkoxy groups are optionally substituted.
  • cycloalkyl alone or in combination, means an alkyl radical which contains at least one carbon ring. These groups may be monocyclic, bicyclic or tricyclic. Unless otherwise indicated a cycloalkyl contains from 3 to 12 carbons and the carbon ring contains 3-10 carbons and more preferably 3-7 carbon atoms. Examples of such cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups are optionally substituted.
  • alkenyl refers to a straight-chain or branched-chain mono-,di- or poly-unsaturated aliphatic hydrocarbon radical containing one or more double bonds and the specified number of carbon atoms, or where no number is specified, preferably from 2-10 carbon atoms, more preferably, from 2-6 carbon atoms and also from 2-4 carbon atoms. Unless specifically stated, all isomers of the given number of carbon atoms are included.
  • alkenyl radicals include, but are not limited to, ethenyl, E- and Z-propenyl, isopropenyl, E- and Z-butenyl, E- and Z-isobutenyl, E- and Z-pentenyl, E- and Z-hexenyl, E,E-, E,Z-, Z,E- and Z,Z hexadienyl.
  • Alkenyl groups are optionally substituted.
  • alkenoxy refers to an -O-alkenyl group, where alkenyl is defined above. Alkenoxy groups are optionally substituted.
  • cycloalkenyl means an alkyl radical which contains at least one carbon ring and at least one double bond. These groups may be monocyclic, bicyclic or tricyclic. Unless otherwise indicated a cycloalkyl contains from 3 to 12 carbons and the carbon ring contains 3-10 carbons and more preferably 3-7 carbon atoms. Examples of such cycloalkenyl groups include cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. Cycloalkenyl groups are optionally substituted.
  • alkynyl refers to a straight-chain or branched-chain aliphatic hydrocarbon radical containing one or more triple bonds and the specified number of carbon atoms, or where no number is specified, preferably from 2-10 carbon atoms, more preferably, from 2-6 carbon atoms and also from 2-4 carbon atoms. Specific alkynyl groups contain one triple bond or two triple bonds. Unless specifically stated, all isomers of the given number of carbon atoms are included. Examples of alkynyl radicals include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc. Alkynyl groups are optionally substituted.
  • alkynoxy refers to an -O-alkynyl group, where alkynyl is defined above. Alkynoxy groups are optionally substituted.
  • An acyl group is an R'-CO- group where R' in general is a hydrogen, an alkyl, alkenyl or alkynyl, aryl, heterocyclyl, or heteroaryl group as described herein.
  • R' is a substituted methylene group.
  • acyl groups have 1 -20, 1 - 12, or 1 -6 carbon atoms and optionally 1 -3 heteroatoms, optionally one double bond or one triple bond.
  • R is a C1 -C6 alkyl, alkenyl group cyclic configuration or a combination thereof, attached to the parent structure through a carbonyl functionality.
  • Examples include acetyl, benzoyl, propionyl, isobutyryl, or oxalyl.
  • the R' group of an acyl group is optionally substituted as described herein. When R' is hydrogen, the group is a formyl group.
  • An acetyl group is a Chh- CO- group.
  • Another exemplary acyl group is a benzyloxy group.
  • An acylalkoxy group is an alkoxy group as defined above substituted with an acyl group as defined above, e.g., -OCH2-COR.
  • An alkyl group substituted with an acylalkoxy group is an acylalkoxyalkyl group.
  • An acyloxy group is an acyl group as defined above bonded to an oxygen, e.g., -O-COR.
  • An alkyl group substituted with an acyloxy group is an acyloxyalkyl group.
  • An acylamino group is an R'-CO-NH- group, where R' is as defined for the acyl group above.
  • R' is as defined for the acyl group above.
  • a number of acylamino groups are known in the art as suitable for use in cephem and penem antibiotics.
  • An amino group is -NH2.
  • An alkylamino group is -NHR', where R' is an alkyl group, preferably which is a C1 -C4 alkyl or a C1 -C3 alkyl.
  • An alkylamino group is -NR2', where R' is an alkyl group, preferably which is a C1 -C4 alkyl or a C1 -C3 alkyl.
  • Amino, alkylamino and dialkyl amino groups can be protonated or quaternized, for example, with bonding of another alkyl group resulting in a positively charged amino, alkyl amino, dialkyamino, trialky amino or a quaternary amino group.
  • Certain compounds herein have cyclic amino groups where the nitrogen is in a ring.
  • the ring may otherwise contain only carbon ring atoms or the ring may contain an additional heteroatom, such as 0, N or S.
  • the ring may be saturated (containing no double bonds), partially unsaturated or fully unsaturated.
  • the ring N can be protonated or alkylated (with a C1 -C4 alkyl group) to form a positively charged ring amino group.
  • aryl refers to a carbocyclic aromatic radical (such as phenyl or naphthyl) containing the specified number of carbon atoms. If not specified aryl groups contain from 6-15 carbon atoms, preferably from 6-10 carbon atoms, and particularly contain from 6-10 ring carbons. Aryl groups unless otherwise stated are optionally substituted among others with one or more substituents selected from alkyl, alkoxy, nitro, halogen, (for example, chloro), amino, alkyl amino, dialkylamino, carboxylate and hydroxy. In specific embodiments, aryl groups are optionally substituted phenyl groups.
  • Aryl groups may contain two rings that are fused (naphthyl) or two rings which are bonded together by a C-C bond (biphenyl).
  • aryl groups include, among others, phenyl, p-tolyl, 4-hydroxyphenyl, 1 -naphthyl, 2-naphthyl, indenyl, indanyl, azulenyl, fluorenyl, and anthracenyl.
  • arylalkyl refers to an alkyl group as described above which is substituted with an aryl group as defined above.
  • the aryl group is an optionally substituted phenyl, naphthyl or biphenyl group.
  • the arylalkyl is a phenalkyl group.
  • Specific phenalkyl groups are benzyl and phenethyl groups.
  • heterocyclyl or“heterocyclic” refers to monoradical having a ring of a specified number of ring atoms, where the ring atoms include one or more heteroatoms (N, O, S) or heteroatom groups (e.g., -NH-, or -N(alkyl)-). More specifically, the term includes groups having a stable 3-7 membered monocyclic heterocyclic ring or an 8-1 1 membered bicyclic heterocyclic ring.
  • a number of heterocyclic groups are exemplified in the specification.
  • Each heterocycle consists of one or more carbon atoms and from one to four heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur and oxidized forms thereof.
  • Heterocycles include 5-7 membered monocyclic heterocycles and 8-10 membered bicyclic heterocycles.
  • heteroaryl refers to a group having at least one aromatic ring wherein the ring contains at least one heteroatom or heteroatom group, as defined above. More specifically, the term refers to stable 5-6 membered monocyclic or 8-1 1 membered bicyclic aromatic heterocycles where heterocycles is as defined above.
  • Non-limiting examples of such groups include imidazolyl, quinolyl, isoquinolyl, indolyl, indazolyl, pyridazyl, pyridyl, pyrrolyl, pyrazolyl, pyrazinyl, quinoxolyl, pyranyl, pyrimidinyl, furyl, thienyl, triazolyl, thiazolyl, carbolinyl, tetrazolyl, benzofuranyl, thiamorpholinyl sulfone, oxazolyl, benzoxazolyl, benzimidazolyl, benzthiazolyl, oxopiperidinyl, oxoppyrrolidinyl, oxoazepinyl, azepinyl, isoxazolyl, isothiazolyl, furazanyl, thiazolyl, thiadiazolyl, and oxathiolyl.
  • a carbocyclyl group is a group having one or more saturated or unsaturated carbon rings.
  • Carbocyclyl groups for example, contain one or two double bonds.
  • One or more carbons in a carbocyclic ring can be -CO- groups.
  • Carbocyclyl groups include those having 3-12 carbon atoms, and optionally replacing 1 or 2 carbon atoms with a - CO- group and optionally having 1 , 2 or 3 double bonds.
  • Carbocyclyl groups include those having 5-6 ring carbons.
  • Carbocyclyl groups can contain one or more rings each of which is saturated or unsaturated.
  • Carbocyclyl groups include bicyclic and tricyclic groups. Preferred carbocyclic groups have a single 5- or 6- member ring.
  • Carbocyclyl groups are optionally substituted as described herein. Specifically, carbocyclic groups can be substituted with one or more alkyl groups. Carbocyclyl groups include among others cycloalkyl and cycloalkenyl groups.
  • a styryl group or moiety is a group or moiety in which a vinyl (ethylene) group is bonded to a phenyl ring.
  • the group or moiety may be substituted with one or more non-hydrogen substituents on the phenyl ring or most generally on the vinyl moiety.
  • the styryl group can be cis or trans or with respect to the monobactam ring or a mixture of cis and trans isomers.
  • the cis and trans configurations can also be designated the E or Z configuration (or a mixture of E/Z configurations), where one of ordinary skill in the art understand how to name E and Z configurations for a given structure.
  • the E and Z configurations are exemplified in styryl groups having formulas:
  • R5-R9 are optional substituents as defined for the B ring herein and one of R5-R9 is -CH2-X.
  • the vinyl group can be written as a cross double bond to indicate that the double bond is a mixture of the E and Z configurations (cis and trans isomers).
  • Compounds herein can be a mixture of isomers having the E or Z configuration at the styryl group.
  • monobactams herein include variants of the styryl group in which one-four of the carbons of the phenyl ring are replaced with nitrogens, for example, a -vinylpyridyl group.
  • heteroaryl styryl variants can also be in the cis or trans configuration (or E and Z configurations) with respect to the monobactam ring.
  • These vinylheteroaryl groups can be in the cis or trans configuration or can be a mixture of cis/trans configurations, e.g., the compound would be a mixture of cis/trans (or E/Z isomers).
  • ring atoms are numbered as indicated and T is -S-, -SO-, -SO2-, -CHR17-, or -0-. This numbering is used throughout for labeling of ring substituents. It will be appreciated by one of ordinary skill in the art that different ring numbering conventions may be employed in the art. [0270] The term“penem” refers generally to the ring system:
  • ring atoms are numbered as indicated and U is -S-, -CHR17- or -0-. This numbering is used throughout for labeling of ring substituents. It will be appreciated by one of ordinary skill in the art that different ring numbering conventions may be employed in the art. It is noted that the term penem is used generically herein. The term penem is also used in the art to refer to compounds of this ring system where U is -S-.
  • Carboxylate esters include those where E is an optionally substituted C1 -C6 or C1 -C3 alkyl (optional substation includes among others substitution with one or more halogen), an optionally substituted phenyl or phenyalkyl group (where substitution includes among others ring substitution with one or more halogen or with a nitro group).
  • Carboxylate esters further include those in which E is an alkoxyalkyl group, more specifically where the alkoxy and the alkyl group is C1 -C6 or C1 -C3 alkoxy and C1 -C6 or C1 -C3 alkyl, e.g.
  • Carboxylate esters include those where E is an acylalkoxyalkyl group, more specifically where the acyl, alkoxy and alkyl groups are C1 - C6 or C1 -C3 acyl, alkoxy or alkyl groups.
  • R1-CO-NH- in formulas herein is an acylamino group.
  • R1-CO-NH- can be any pharmaceutically acceptable A such group that is known in the art to be compatible with a cephem antibiotic.
  • a number of examples of Ri useful groups are provided in the specification. Additional useful Ri groups are known in the art.
  • useful Ri groups are substituted methylene groups.
  • useful Ri groups are substituted oximes.
  • useful Ri groups are substituted vinyl ether groups.
  • Compound 2 is prepared by a literature procedure [25] More specifically, 4-bromobenzyl alcohol 1 (1 .400 g, 7.485 mmol, 1.000 equiv) is dissolved in anhydrous THF (21 ml_, 0.35 M) at room temperature(rt), under anhydrous conditions. At - 78 °C, a 2.5 M solution of n-BuLi in hexanes (6.14 ml_, 15.3 mmol, 2.05 equiv) is added dropwise to the THF solution with stirring. The resulting mixture solidified after addition of n-BuLi.
  • the aqueous layer is separated from the organic layer and is extracted with AcOEt (ca. 30 ml_).
  • the combined organic layers are dried over magnesium sulfate, filtered and evaporated under reduced pressure.
  • the colorless oil obtained is chromatographed on silica gel (Biotage SNAP, 50 g) eluting with 0-20% AcOEt/Hexanes.
  • Product 2 is obtained as colorless oil (2.522 g, 6.350 mmol, 85% yield).
  • trifluoromethanesulfonyl anhydride (0.926 ml_, 5.514 mmol, 1.200 equiv) is added dropwise.
  • the resulting white suspension is stirred at - 20 °C for 1 hour.
  • the suspension is diluted with DCM (300 ml_) and the resulting organic layer is mixed with 0.25 N HCI (80 ml_) and then with water (150 ml_).
  • the washed organic layer is dried over magnesium sulfate, filtered and solvent is removed under reduced pressure.
  • the product 4 is obtained as pale-orange solid (2.801 g, 4.428 mmol, 96% yield). This product is used without further purification in the next step.
  • the product is further purified by a second column chromatography on silica gel (Biotage SNAP 50 g) eluting with 20-60% AcOEt/Hexanes.
  • Product 5 is obtained as orange solid (1 .275 g, 2.159 mmol, 49% yield).
  • alcohol 5 (1 .270 g, 2.150 mmol, 1 .000 equiv) is dissolved in anhydrous DCM (18 ml_, 0.12 M).
  • methanesulfonyl chloride (0.250 ml_, 3.23 mmol, 1 .50 equiv) is added dropwise, followed by dropwise addition of /V-methylmorpholine (NMM) (0.307 ml_, 2.80 mmol, 1 .30 equiv).
  • NMM /V-methylmorpholine
  • the resulting pale-yellow suspension is stirred between - 5 °C and 0 °C for 1 .5 hours.
  • anhydrous methanol (6.73 ml_, 166 mmol, 100 equiv) is added dropwise to the suspension and the resulting solution is stirred at 0 °C for 15 minutes and then at rt for 10 minutes.
  • the solution is then diluted in DCM (80 ml_) and saturated sodium bicarbonate solution (80 ml_) is added carefully.
  • the separated organic layer is washed with dilute (ca. 0.1 M) sodium bicarbonate solution, dried over magnesium sulfate, filtered and solvent is evaporated until 10-15 mL of the solution of free amine 7 remained.
  • the off-white solid obtained is purified by reverse phase chromatography (Biotage C18 60 g) eluting with 5-100% ACN/H2O.
  • the product 10 is obtained as off-white solid (0.797 g, 0.883 mmol, 53% yield).
  • the orange solid obtained is triturated three times in 50/50 v/v diethyl ether/hexanes (3 x 4 ml_). Remaining traces of solvent are evaporated. The product 11 is obtained as pale-orange solid (0.084 g, quantitative yield), which is used without further purification in the next step.
  • the resulting cloudy dark green mixture is immersed in a preheated oil bath (50 °C) and is stirred at 50 °C for 1 .5 hours. The heating is stopped and 10 drops of TFA are added, followed by ca. 2 ml_ of water. This solution is charged onto a reverse phase column (Biotage C18 60 g) and eluted with a 5-100% ACN/H2O gradient. Fractions containing the desired product were lyophilized and the product 15 is obtained as a fluffy white solid (0.054 g, 0.076 mmol, 45% yield).
  • carboxylic acid 16 (0.102 g, 0.340 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (2.3 ml_, 0.15 M) at rt.
  • methanesulfonyl chloride (0.034 ml_, 0.44 mmol, 1 .3 equiv) is added dropwise, followed by N- methylmorpholine (0.037 ml_, 0.34 mmol, 1.0 equiv).
  • the homogeneous mixture containing compound 17 is stirred at 0 °C for 5 minutes and then at rt for 25 minutes.
  • the resulting yellow oil is purified by normal phase chromatography (Biotage SNAP 25 g) eluting with 20-60% AcOEt/Hexanes.
  • the product 18 is obtained as off-white solid (0.182 g, 0.235 mmol, 80% yield).
  • Compound 23 is prepared by a literature procedure (U.S. Patent 5,283,242). More specifically, under anhydrous conditions, (6-bromopyridin-3- yl)methanol 22 (from Combi-Blocks) (1.000 g, 5.319 mmol, 1.000 equiv) is dissolved in anhydrous DMF (8 ml_, 0.7 M) at rt. At rt, triethylamine (0.771 ml_, 5.53 mmol, 1.04 equiv) is added slowly.
  • fe/f-butyldimethylsilyl chloride (0.834 g, 5.53 mmol, 1.04 equiv) is added as a solid and the remaining material in the weighing vial is transferred as a solution in DMF (2 ml_). The cloudy suspension is stirred at rt for 1 hour and 10 minutes. Diethyl ether (50 ml_) is added as well as water (50 ml_). Layers were separated and the organic layer is washed with water (40 mL), dried over magnesium sulfate and filtered. Solvent is evaporated under reduced pressure.
  • substrate 23 (1 .35 g, 4.47 mmol, 1 .00 equiv) is dissolved in anhydrous THF (1 1 mL, 0.40 M).
  • anhydrous THF 1 1 mL, 0.40 M.
  • n-BuLi in hexanes (1 .88 mL, 4.69 mmol, 1 .05 equiv) is added dropwise.
  • the solution turns orange and then dark green.
  • the solution is then stirred at - 78 °C for 1 hour.
  • the obtained dark brown oil is purified by column chromatography on silica gel (Biotage SNAP, 100 g) eluting with 0-100% AcOEt/Hexanes.
  • the obtained product is purified by a second column chromatography on silica gel (Biotage SNAP, 50 g) eluting with 0-80% AcOEt/Hexanes.
  • the desired product 25 is obtained as red solid, contaminated by some by-product (1 .29 g). The product is used without further purification in the next step.
  • TBS-protected substrate 25 (not pure, 1 .069 g, 1 .514 mmol, 1 .000 equiv) is dissolved in anhydrous THF (5.0 ml_, 0.30 M) at rt. This solution is cooled to 0 °C and acetic acid (0.87 ml_, 15 mmol, 10 equiv) is added. At 0 °C, the 1 .0 M TBAF solution in TFIF (4.54 ml_, 4.54 mmol, 3.00 equiv) is added dropwise. The resulting solution is stirred at 0 °C for 3 hours and 10 minutes.
  • anhydrous methanol (2.67 mL, 65.9 mmol, 100 equiv) is added dropwise and the solution is stirred between 0 °C and 10 °C for 20 minutes.
  • This reaction mixture is diluted in DCM (50 mL) and saturated sodium bicarbonate solution (40 mL) is carefully added. Layers are separated and the aqueous wash is extracted using DCM (30 mL). The combined organic extracts are washed with sat. sodium bicarbonate solution (40 mL), dried over magnesium sulfate, filtered and solvent is evaporated until ca. 10 of solution remained.
  • a solution of acid chloride is prepared as follows:
  • phosphorus pentachloride (0.189 g, 0.906 mmol, 1 .20 equiv) is suspended in anhydrous DCM (3.8 mL, 0.20 M) at rt. The resulting white suspension is stirred at 0 °C for 10 minutes. At 0 °C, the carboxylic acid 8 (0.324 g, 0.755 mmol, 1 .00 equiv.) is added in one portion. The heterogeneous mixture of acid chloride is stirred, between at 0 °C for 1 hour.
  • This material is further purified by normal phase chromatography on silica gel (Biotage SNAP, 25 g), eluting with 10-80% AcOEt/Hexanes.
  • the desired product 29 is separated from its double-bond migration isomer and it is obtained as off-white solid (0.335 g, 0.371 mmol, 56% yield).
  • substrate 29 (0.335 g, 0.371 mmol, 1.00 equiv) is dissolved in anhydrous DCM (3.7 ml_, 0.10 M).
  • anisole (0.201 ml_, 1 .85 mmol, 5.00 equiv) is added slowly, quickly followed by slow addition of TFA (5.3 ml_, 0.070 M).
  • TFA 5.3 ml_, 0.070 M
  • the pale-yellow solution is stirred at 0 °C for 2 hours and then at rt for 1 .5 hours. This reaction mixture is then evaporated to dryness.
  • the orange oil obtained is triturated in diethyl ether (4 x 4 ml_).
  • 2-methyl-2/-/-indazol-1 -ium bromide 14 (0.050 g, 0.21 mmol, 12 equiv), potassium carbonate (0.019 g, 0.14 mmol, 8.0 equiv) and substrate 30 (0.010 g, 0.017 mmol, 1 .0 equiv) are suspended in anhydrous DMF (0.2 ml_, 0.1 M).
  • Potassium iodide (0.006 g, 0.03 mmol, 2 equiv) is added and the resulting cloudy purple mixture is immersed in a preheated oil bath (40 °C) and is stirred at 40 °C for 1 .5 hours.
  • Compound 34 is prepared by a literature procedure (WO 2017/16261 1 , PCT/EP2017/056599). More specifically, under anhydrous conditions, (5- bromopyridin-2-yl)methanol 33 (from Combi-Blocks) (2.000 g, 10.64 mmol, 1.000 equiv) is dissolved in anhydrous DMF (19 mL, 0.56 M) at rt. At rt, imidazole (1 .09 g, 16.0 mmol, 1 .50 equiv) is added in one portion.
  • fe/f-butyldimethylsilyl chloride (1.92 g, 12.8 mmol, 1 .20 equiv) is added as a solid and the remaining material in the weighing vial is transferred as a solution in DMF (2 mL). The resulting clear yellow solution is stirred at rt for 1 hour and 45 minutes. AcOEt (70 mL) and water (70 mL) are then added. Layers are separated and the organic layer is washed with water (40 mL), then with brine. It is then dried over sodium sulfate and filtered. Solvent is evaporated under reduced pressure.
  • the pale-yellow oil obtained is chromatographed on silica gel (Biotage SNAP, 100 g), eluting with 100% DCM.
  • the desired product 34 is isolated as colorless oil (3.17 g, 10.5 mmol, 99% yield).
  • substrate 34 (1 .50 g, 4.96 mmol, 1 .00 equiv) is dissolved in anhydrous THF (12 ml_, 0.40 M).
  • anhydrous THF (12 ml_, 0.40 M).
  • n-BuLi in hexanes (2.08 ml_, 5.21 mmol, 1 .05 equiv) is added dropwise.
  • the solution turned to orange and then to dark brown.
  • the solution is stirred at - 78 °C for 40 minutes.
  • reaction solution is left at 4 °C, without stirring for 3 days.
  • the reaction solution is then transferred to a larger flask and water (100 ml_) is gradually added to the dark brown solution. Orange precipitate appears while a small exotherm is observed. The precipitate is collected by filtration on sintered glass. The orange precipitate is washed with water (3 x 100 ml_) and dried on a sintered glass funnel. The solid (hydrated) is dried under high vacuum for 24 hours.
  • Product 36 is obtained as pale yellow solid (5.08 g, 9.79 mmol, 98% yield). This product is used without further purification in the next step.
  • the chloride 36 (1 .00 g, 1 .93 mmol, 1 .00 equiv) is introduced into a flask.
  • a solution of stannane 35 (1 09g, 2.12 mmol, 1 .10 equiv) in anhydrous NMP (14 mL, 0.15 M) is added and nitrogen is bubbled into the resulting orange solution.
  • zinc chloride (0.53 g, 3.9 mmol, 2.0 equiv
  • tri(2-furyl)phosphine (0.179 g, 0.771 mmol, 0.40 equiv), while nitrogen is still being bubbled into the solution.
  • tris(dibenzylideneacetone)dipalladium (0.353 g, 0.385 mmol, 0.20 equiv) is added in one portion.
  • the resulting deep green/brown mixture is immersed in a preheated oil bath (70 °C) and is stirred at 70 ° for 1 .5 hours. Heating is then stopped, and the reaction mixture is diluted in AcOEt (60 mL). This organic layer is washed with sat. ammonium chloride solution (3 x 50 mL) and then with brine. Solvent is evaporated under reduced pressure. The resulting brown oily film is taken up in ACN (50 mL).
  • TBS-protected substrate 37 from step D (0.765 g, 1 .08 mmol, 1 .00 equiv) is dissolved in anhydrous THF (3.1 mL, 0.35 M) at rt. This solution is cooled to 0 °C and acetic acid (0.62 mL, 1 1 mmol, 10 equiv) is added. At 0 °C, a 1 .0 M TBAF solution in THF (1 .84 mL, 1 .84 mmol, 1 .70 equiv) is added dropwise. The resulting solution is stirred at 0 °C for 5 hours and 10 minutes. AcOEt (50 mL) and sat.
  • phosphorus pentachloride (0.192 g, 0.921 mmol, 1.15 equiv) is suspended in anhydrous DCM (3.2 mL, 0.25 M) at rt. The resulting white suspension is stirred at 0 °C for 15 minutes. At 0 °C, the carboxylic acid 8 (0.344 g, 0.801 mmol, 1.00 equiv) is added in one portion. The heterogeneous mixture is stirred, at 0 °C for 20 minutes.
  • substrate 41 (0.380 g, 0.421 mmol, 1.00 equiv) is dissolved in anhydrous DCM (4.2 ml_, 0.10 M).
  • anisole (0.18 ml_, 1 .7 mmol, 4.0 equiv) is added slowly, quickly followed by slow addition of TFA (5.3 ml_, 0.080 M).
  • the yellow solution is stirred at 0 °C for 3 hours and 40 minutes and then at rt for 1 hour and 20 minutes. This reaction mixture is then evaporated to dryness.
  • the orange oil obtained is triturated in diethyl ether (3 x 4 ml_).
  • Potassium iodide (0.068 g, 0.412 mmol, 1 .00 equiv) is added and the suspension is stirred at rt for 1 hour and 50 minutes. TFA (1 .5 ml_, 20 mmol, 48 equiv) is then added, followed by ca. 2.5 ml_ of water. The resulting solution is chromatographed on a reverse phase column (Teledyne GOLD C18, 60 + 60 g), eluting with a 0-100% ACN/H2O gradient (+ 0.1 % TFA). Fractions containing the desired product are combined and lyophilized. The desired product 43 is obtained as off-white solid (0.249 g, 0.352 mmol, 86% yield).
  • (4-aminophenyl)methanol 44 (1 .00 g, 8.13 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (20 mL, 0.40 M).
  • fe/f-butyldimethylsilyl chloride (1 .47 g, 9.76 mmol, 1 .20 equiv) is added and the resulting solution is stirred at rt for 5 minutes.
  • Triethylamine (2.3 mL, 16 mmol, 2.0 equiv) is added at rt and the reaction mixture is stirred at rt for 4 hours.
  • the reaction mixture is diluted in AcOEt and this organic layer is washed with water, sat.
  • (S)-2-amino-3-hydroxypropanoic acid 46 (2.00 g, 19.0 mmol, 1.00 equiv) and sodium carbonate (2.00 g, 19.0 mmol, 1.00 equiv) are dissolved in water (20 mL, 0.95 M).
  • /V-Ethoxycarbonylphthalimide 47 (4.18 g, 19.0 mmol, 1.00 equiv), previously ground to a fine powder, is added at rt and the resulting suspension is stirred at rt until no more solid can be seen.
  • the pH is adjusted to 1 by adding a 1 N HCI, at 0 °C. Some precipitate then appeared in the solution. This aqueous layer is extracted with AcOEt.
  • the organic extract is evaporated under reduced pressure and oil is obtained.
  • the oil is taken up in the minimum amount of DCM (10 mL) and toluene (30 mL) is added. Crystallization occurred over 3 days and the resulting white solid is filtered off, washed with toluene and hexanes and dried under vacuum.
  • the desired product 48 is obtained as white solid (1.9 g, 8.1 mmol, 42% yield).
  • the washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the residue is purified by column chromatography on silica gel.
  • the desired phenyl monobactam product 50 is obtained as white solid (1.04 g), which is contaminated with some DIAD by-product. This material is used without further purification in the next step.
  • Phenyl monobactam substrate 50 (1 .04 g, 2.38 mmol, 1 .00 equiv) is suspended in anhydrous EtOH (10 mL, 0.24 M). Ethylenediamine 51 (0.476 mL, 7.15 mmol, 3.00 equiv) is added at rt. THF (5 mL, 0.5 M) is added to dissolve solids present in the suspension. The resulting solution is stirred overnight at rt. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NH4CI, sat. aqueous NaHC03 and then with brine.
  • carboxylic acid 8 (0.547 g, 1 .27 mmol, 1 .05 equiv) is dissolved in anhydrous DMA (6 ml_, 0.2 M).
  • methanesulfonyl chloride (0.102 ml_, 1 .30 mmol, 1 .05 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.180 ml_, 1.30 mmol, 1.05 equiv).
  • the resulting pale yellow suspension is stirred at 5 °C for 1 hour.
  • the washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the residue is purified by column chromatography on silica gel, eluting with 0-25% AcOEt/DCM gradient. The desired compound 57 is obtained.
  • the latter is dissolved in a mixture of triethylsilane (0.025 mL), water (0.03 mL) and TFA (1.0 mL), at 0 °C.
  • the resulting reaction mixture is stirred at 0 °C for 3.5 hours and then 0.5 hour at rt.
  • Solvent is evaporated under reduced pressure and the residue is triturated twice in diethyl ether.
  • the desired compound 58 (478) is obtained as off-white solid (0.012 g, 0.02 mmol, 75% yield over 2 steps).
  • the resulting solution is stirred at rt for 2 hours and then at 50 °C for 1 hour.
  • the reaction mixture is purified by C18 reverse phase column chromatography. Fractions containing the desired intermediate are combined and lyophilized.
  • the intermediate is dissolved in TFA (1.0 ml_, 0.01 M) at rt and the resulting solution is stirred at rt for 1 .5 hours. Solvent is evaporated under reduced pressure and the residue is triturated twice in diethyl ether.
  • the solid obtained is purified by C18 reverse phase column chromatography. Fractions containing the desired product are combined and lyophilized.
  • the desired product 61 (481 ) is obtained as white solid (0.005 g, 0.01 mmol, 12% yield over 2 steps).
  • Compound 48 is prepared from (S)-2-amino-3-hydroxypropanoic acid 46 following the procedure described in Example 4, Procedure B.
  • the resulting solution is transferred via cannula to a stirred suspension of 0-(diphenylphosphinyl)hydroxylamine 63 (2.31 g, 9.90 mmol, 1 .00 equiv) in anhydrous THF (20 mL, 0.50 M), at - 78 °C.
  • the resulting mixture is stirred for 1 hour at - 78 °C and then warmed to rt and stirred at rt for 2 hours.
  • the reaction mixture is diluted in AcOEt (200 mL). This organic layer is washed with 10% aqueous citric acid (3 x 80 mL).
  • the combined aqueous washes are brought to pH 8-9 by addition of sodium carbonate, and are then extracted using DCM.
  • the DCM extract is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the desired compound 64 is obtained as colourless oil (1 .84 g, 6.89 mmol, 70% yield).
  • the product compound is dissolved in diethyl ether (10 mL) and left at - 20 °C for one night. Some triphenylphosphine oxide crystallized out of the solution and is removed by filtration. The filtrate still contained triphenylphosphine oxide. The filtrate is adsorbed onto silica gel prior to normal phase chromatography eluting with a AcOEt/hexanes gradient. The desired compound 66 is obtained (mixture of 2 diastereoisomers, 0.780 g, 1 .67 mmol, 33% yield).
  • Compound 68 is prepared by a literature procedure (Wang et al. (2017) Org. Lett. 19(3):632-635). More specifically, under inert atmosphere, terephthalaldehyde 67 (10.0 g, 74.6 mmol, 1 .00 equiv) is dissolved in anhydrous THF (124 mL, 0.60 M) and ethanol (41 mL, 1.8 M). At 0 °C, sodium borohydride (0.705 g, 18.6 mmol, 0.25 equiv) is added in small portions, over 20 minutes. The resulting mixture is stirred at 0 °C for 2.5 hours.
  • More sodium borohydride (0.039 g, 1 .0 mmol, 0.014 equiv) is added at 0 °C.
  • the resulting mixture is stirred at 0 °C for 2 hours.
  • the reaction is quenched by addition of aqueous 2M HCI (10 mL).
  • the resulting yellow solution, containing white solids is kept at 0 °C for the night.
  • Solvent is evaporated under reduced pressure.
  • Water (150 mL) is then added to the residue and the product is extracted with AcOEt (2 x 100 mL). Organic extracts are combined and washed with brine. The organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the residue is purified by column chromatography on silica gel (Biotage SNAP, 100 g) eluting with 0-50% AcOEt/hexanes gradient.
  • the desired product 68 is obtained as a white solid (6.68 g, 49.1 mmol, 66% yield).
  • aldehyde 69 (0.438 g, 1.75 mmol, 1.20 equiv) and phosphonate 66 (0.680 g, 1.46 mmol, 1.00 equiv) are dissolved in anhydrous THF (10 ml_ 0.15 M).
  • sodium hydride (60% suspension in mineral oil) (0.064 g, 1.6 mmol, 1.1 equiv) is added.
  • the resulting mixture is stirred at 0 °C for 0.5 hour and then at rt for 2 hours.
  • the reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NFUCI and then with brine.
  • the washed organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the yellow oil obtained is purified by column chromatography on silica gel.
  • the desired compound 70 (pure trans double bond) is obtained as white solid (0.300 g, 0.533 mmol, 37% yield).
  • carboxylic acid 8 (0.209 g, 0.487 mmol, 1 .05 equiv) is dissolved in anhydrous DMA (3.0 ml_, 0.16 M).
  • methanesulfonyl chloride (0.040 ml_, 0.51 mmol, 1 .1 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.071 ml_, 0.51 mmol, 1 .1 equiv).
  • the resulting pale yellow suspension is stirred at 5 °C for 1 hour.
  • the latter is dissolved in anhydrous DCM (0.5 mL, 0.05 M).
  • TFA 0.5 mL, 0.05 M
  • Solvents are evaporated under reduced pressure and the residue is triturated twice in diethyl ether.
  • the desired compound 76 (GL340) is obtained as white solid (0.013 g, 0.020 mmol, 73% yield over 2 steps).
  • reaction solution is purified by C18 reverse phase chromatography. Fractions containing the desired intermediate are combined and lyophilized.
  • TFA 0.5 ml_, 0.05 M
  • the resulting solution is stirred at rt for 1 .5 hours.
  • Solvent is evaporated under reduced pressure.
  • the residue is triturated twice in diethyl ether.
  • the desired product 77 (491 ) is obtained as white solid (0.005 g, 0.008 mmol, 14% yield over 2 steps).
  • the reaction mixture is purified by C18 reverse phase chromatography. Fractions containing the desired intermediate are combined and lyophilized.
  • TFA 0.5 ml_, 0.05 M
  • the resulting solution is stirred at rt for 1 .5 hours.
  • Solvent is evaporated under reduced pressure.
  • the residue is triturated twice in diethyl ether.
  • the desired product 79 (495) is obtained as white solid (0.007 g, 0.01 1 mmol, 16% yield over 2 steps).
  • Compound 48 is prepared from (S)-2-amino-3-hydroxypropanoic acid 46 following the procedure described in Example 4, Procedure B.
  • the reaction mixture is diluted in AcOEt (25 mL) and this organic solution is washed with water (20 mL) and sat. aqueous NaHCCb (20 mL). Solid sodium bicarbonate is added to the aqueous wash until saturated. This aqueous layer is extracted with AcOEt (20 mL). The organic extracts are combined and dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure.
  • Solvent is evaporated under reduced pressure and the residue is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with a 0-40% AcOEt/DCM gradient. The fractions containing the desired compound are combined and solvent is evaporated under reduced pressure. The syrup obtained is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with 0-40% AcOEt/DCM. The desired product 82 is obtained as a colourless oil (0.124 g, 0.435 mmol, 34% yield over 2 steps).
  • the washed organic layer is then dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the white solid obtained is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with 0-10% MeOH/DCM.
  • the desired product 83 is obtained as a white solid (0.069 g, 0.32 mmol, 73% yield).
  • tert- butyl 2-(triphenylphosphoranylidene)acetate 84 (from Oakwood Chemicals) (0.508 g, 1.35 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (9.0 ml_, 0.15 M). At O °C, bromide (0.076 ml_, 1 .5 mmol, 1.1 equiv) is added dropwise. The resulting yellow solution is stirred at 0 °C for 45 minutes. The reaction mixture is diluted in DCM (15 ml_) and this organic layer is washed with water (15 ml_) and then sat. aqueous NaHCCb (20 ml_).
  • carboxylic acid 8 (0.037 g, 0.087 mmol, 1.20 equiv) is dissolved in anhydrous DCM (0.9 ml_, 0.1 M).
  • methanesulfonyl chloride (0.008 ml_, 0.1 mmol, 1.5 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.012 ml_, 0.087 mmol, 1.2 equiv).
  • the resulting pale yellow suspension is stirred at 0 °C for 50 minutes.
  • a solution of substrate 87 (0.023 g, 0.072 mmol, 1.0 equiv) in anhydrous DCM (0.7 ml_, 0.1 M) is prepared. At 0 °C, it is added to the solution of mesylate, followed by /V-methylmorpholine (0.008 ml_, 0.07 mmol, 1 equiv). The resulting cloudy colourless solution is stirred between 0 °C and rt for 3.5 hours. The reaction mixture is diluted in DCM (20 ml_) and this organic layer is washed with sat. aqueous NhUCI (15 ml_).
  • Methanesulfonyl chloride (0.004 ml_, 0.05 mmol, 1 equiv) is added and the reaction mixture is stirred at rt for 3 hours. Solvent is evaporated under reduced pressure. Lithium chloride (0.0060 g, 0.14 mmol, 3.0 equiv) is added to the residue, followed by anhydrous DMF (0.5 mL, 0.1 M). The resulting pale yellow suspension is stirred at rt for 17 hours. The reaction mixture is diluted in AcOEt (20 mL) and this organic layer is washed with sat. aqueous NhUCI (2 x 10 mL).
  • the washed organic layer is then dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure.
  • the residue is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with a 10-40% AcOEt/hexanes gradient.
  • the desired product 89 is obtained as a white solid (0.025 g, 0.033 mmol, 70% yield).
  • Potassium iodide (0.003 g, 0.02 mmol, 0.5 equiv) is added and the suspension is stirred at 50 °C for 20 more minutes.
  • the reaction mixture is cooled to rt and is purified by reverse phase column chromatography (Teledyne GOLD C18, 12 g) eluting with 5-100% ACN/H2O (+ 0.1 % TFA gradient). Fractions containing the desired product are combined and lyophilized.
  • the desired product 90 is obtained as white solid (0.024 g, 0.027 mmol, 82% yield).
  • triethylamine (0.208 ml_, 1.50 mmol, 0.960 equiv) is added, followed by dropwise addition of trifluoromethanesulfonic anhydride (from Sigma Aldrich) (0.252 ml_, 1 .50 mmol, 0.960 equiv).
  • the resulting reaction mixture is stirred at -78 °C for 0.5 hour and then triethylamine (0.239 ml_, 1 .71 mmol, 1 .10 equiv) is added, followed by dropwise addition of triethylsilyl trifluoromethanesulfonate (0.39 ml_, 1 .71 mmol, 1 .1 equiv).
  • the resulting reaction mixture is warmed to 30 °C and stirred at this temperature for 1 .5 hours.
  • the reaction mixture is cooled to rt and is diluted in AcOEt (10 ml_). This mixture is filtered to remove any solids, and additional AcOEt (15 ml_) is added. The filtrate is washed with water (15 ml_) and the aqueous wash is extracted with AcOEt (6 ml_). The combined organic extracts are dried over sodium sulfate, filtered and then concentrated under reduced pressure. The residue is purified by column chromatography on silica gel (Biotage SNAP, 50 g) eluting with 0-50% AcOEt/DCM.
  • the filtrate is concentrated under reduced pressure.
  • the residue is purified by reverse phase column chromatography (Biotage C18, 12 g) eluting with 0-100% ACN/H2O. Fractions containing the desired product are combined and lyophilized.
  • the desired product 97 (484) is obtained as a white solid (0.004 g, 0.01 mmol, 13% yield).
  • triethylamine (1 .58 ml_, 1 1 .3 mmol, 0.960 equiv) is added, followed by dropwise addition of trifluoromethanesulfonic anhydride (from Sigma Aldrich) (1 .90 ml_, 1 1 .3 mmol, 0.960 equiv).
  • the resulting reaction mixture is stirred at -78 °C for 0.5 hour and then triethylamine (1 .80 ml_, 13.0 mmol, 1 .10 equiv) is added, followed by dropwise addition of fe/f-butyldimethylsilyl trifluoromethanesulfonate (2.97 ml_, 13.0 mmol, 1 .10 equiv).
  • the resulting mixture is stirred at -78 °C for 0.5 hour.
  • the resulting reaction mixture is warmed to 30 °C and stirred at this temperature for 1 hour.
  • the reaction mixture is cooled to rt and is diluted in AcOEt (100 ml_).
  • This organic layer is washed with water (100 ml_) and the aqueous wash is extracted with AcOEt (100 ml_).
  • the combined organic extracts are dried over magnesium sulfate, filtered and then concentrated under reduced pressure.
  • the residue is purified by column chromatography on silica gel eluting with 0-100% AcOEt/hexanes.
  • the desired product 99 is obtained as pale yellow solid (4.03 g, 6.96 mmol, 59% yield).
  • the biphasic mixture is transferred into a separatory funnel and the mixture is extracted with AcOEt (2 x 50 mL). The organic layer is dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue is purified by column chromatography on silica gel eluting with 0-100% AcOEt/hexanes gradient. The desired product 101 is obtained as pale yellow solid (0.597 g, 1 .24 mmol, 17% yield).
  • the resulting solid is sonicated in diethyl ether.
  • the diethyl ether layer is removed. Remaining traces of solvent are removed by rotatory evaporation and by drying under high vacuum.
  • the desired product 102 is obtained as brown solid (0.046 g, 0.087 mmol, 73% yield).
  • the desired product is extracted from the solid by trituration in MeOH (3 x 5 ml_). The filtrate is concentrated under reduced pressure. The remaining phosphate is removed by solubilizing the product in a minimum of water and passing the solution through an anion exchange resin (IRA-400(CI)) column. The product is eluted with water. Fractions containing the desired product are combined and lyophilized. The desired product 103 (488) is obtained as a yellow solid (0.023 g, 0.059 mmol, 84% yield).
  • MIC Minimum Inhibitory Concentrations
  • the concentration of organisms is adjusted to 0.08 - 0.1 OD6oo (0.5 McFarland units) by dilution with sterile cation-adjusted Meuller-Hinton (CAMH) broth. The suspensions are used within 30 minutes of preparation.
  • Test Compound Stock Solution A stock solution is made of each test compound (by measuring 6.4 mg of the compound on an analytical balance and dissolving it in DMSO (1 ml_) to get a transparent solution. The solutions are stored at -20°C and thawed prior to use.
  • Inoculant suspensions (0.5 pL) are added to the 12 wells of each column. The plates are then covered and incubated at 37°C overnight for 18 hours. For each organism the MIC is determined by identifying the transparent well with the lowest test compound concentration.
  • TEM-26 Group 2be beta-lactamase, hydrolyzes extended spectrum cephalosporins and monobactams), [28];
  • SHV-1 (Class A, Group 2b beta-lactamase, hydrolyzes penicillins and early cephalosporins) [29];
  • DHA-1 (beta-lactamase, intermediate or resistant to all penicillins, alone or in combination with beta-lactamase inhibitors, and to cephalothin, cefuroxime, cefoxitin, ceftazidime, aztreonam, and cefotaxime) [30, 31 ];
  • CTX-M-14 and CTX-M-15 (Class A, Group 2be beta-lactamases, hydrolyzes extended spectrum cephalosporins and monobactams) [32];
  • KPC-2 and KPC-3 (Class A, Group 2f, carbapenemases) [33, 34];
  • NDM-1 Group 3a carbapenemase, broad spectrum divalent Zn ++ metallo- Beta-Lactamase
  • VIM-2 Group 3a carbapenemase, metallo-beta-lactamase [37];
  • IMP-1 Group 3a carbapenemase, metallo-beta-Lactamase.
  • each beta-lactamase enzyme is synthesized and inserted into the pBC-SK(+) plasmid by GenScript, a commercial service.
  • E.coli J53 is transformed with each plasmid via electroporation and resistant transformants are selected on agar plates containing 50 pg/mL ampicillin. The selected transformants are stored at -80 degrees C in 20% glycerol.
  • MIC (pg/mL) results are provided for exemplary compounds of Formula I in Tables 1 -6.
  • Organisms for which MICs are measured are known in the art, can be prepared by well-known methods from available starting materials are available from commercial sources or are clinical isolates.
  • Tables 1 -6 show that the compounds of Formula I (e.g., 501 , 502, 503, 504, 482, 487, 505, 506, and 507) exhibit MIC values that indicate the compounds can inhibit bacterial growth at relatively low concentrations and across vast number of difference organisms.
  • the MIC is defined as the lowest concentration of an antimicrobial ingredient or agent that is bacteriostatic (prevents the visible growth of bacteria). MICs are used to evaluate the antimicrobial efficacy of various compounds by measuring the effect of decreasing concentrations of the antibacterial agent (e.g., GL332) over a defined period in terms of inhibition of microbial population growth.
  • the MBC is the lowest concentration of an antibacterial agent required to kill a bacterium over a fixed.
  • the MBC is complementary to the MIC; whereas the MIC test demonstrates the lowest level of antimicrobial agent that greatly inhibits growth, the MBC demonstrates the lowest level of antimicrobial agent resulting in microbial death.
  • the antimicrobial agent When the MBC to MIC ratio is ⁇ 4, the antimicrobial agent is considered bactericidal. Conversely, when the MBC to MIC ratio is > 8, the antimicrobial agent is considered bacteriostatic. Bactericidal agents kill bacteria, rather than just prevent the growth of bacteria and are therefore, considered to be more potent than bacteriostatic agents.
  • the spectrum of activity was determined for GL-332 by conducting minimal inhibitory concentration (MIC) testing against a range of Gram positive/negative bacteria that included aerobes and anaerobes in medium depleted of iron through treatment with Chelex; in some instances, testing was performed in untreated medium as well.
  • MIC minimal inhibitory concentration
  • MMC minimum bactericidal concentration
  • Test Media Broth microdilution testing of aerobic bacteria was conducted in cation-adjusted Mueller Hinton broth (MHBII). For testing Streptococci, Listeria, Campylobacter and Corynebacterium, lysed horse blood (LHB) was added to MHBII broth at a final concentration of 3%. For testing Haemophilus, Haemophilus test medium (HTM) was used. For testing Neisseria, GC media was used.
  • MHBII Mueller Hinton broth
  • HTM Haemophilus test medium
  • Neisseria Neisseria
  • Broth Microdilution MIC Assay Broth microdilution susceptibility testing in 96-well microplates was conducted. Automated liquid handlers were used to conduct serial dilutions and liquid transfers, including the Biomek 2000, and the Biomek FX (Beckman Coulter, Fullerton, CA). All wells in columns 2 through 12 of a standard 96- well microdilution plate (Costar 3795) were filled with 150 pL of the appropriate diluent. Then, 300 pL of the tested agents were added to the wells of column 1 of the plates at 100X the highest final concentration to be tested. Serial two-fold dilutions were made across the rows through column 1 1 using the Biomek 2000. The wells of column 12 contained no drug and served as the growth control wells. This plate served as the “mother plate” from which MIC assay plates or“daughter plates” were made.
  • the daughter plates were loaded with 188 pL per well of the appropriate medium by hand.
  • the daughter plates were created using the Biomek FX which transferred 2 pL of drug solution from each well of a mother plate to each corresponding well of the daughter plate in a single step.
  • a standardized inoculum of each test organism was prepared per CLSI methods (1 ) to equal a 0.5 McFarland standard, followed by a dilution of 1 :20.
  • the plates were then inoculated with 10 pL of the diluted inoculum using the Biomek 2000 from low to high drug concentration, resulting in a final concentration of approximately 5 x 105 CFU/mL per well.
  • MBC Minimum Bactericidal Concentration
  • the plate was then tilted at a 45°- 90° angle to allow the 10 pL aliquot to track across the agar surface to the opposite side of the plate.
  • the plates were laid flat, allowed to dry at room temperature, then inverted and incubated at 35°C for approximately 24 hr. Colonies were manually counted to determine the viable inoculum count in CFU/mL in each well of the corresponding MIC plate.
  • the sums of the counts for the two spotted aliquots were compared to the values in the appropriate table of rejection values [38] These values were based upon the cell density of the inoculum and the target viable count reduction of 99.9%; if the sum of the colonies was less than or equal to the value in the table, the concentration of drug in the sampled well was considered to be bactericidal [38]
  • the MBC was then defined as the lowest concentration of agent to demonstrate a bactericidal effect.
  • GL332 was observed to have bactericidal activity against several isolates of E. coli, K. pneumoniae and P. aeruginosa.
  • the MIC and MBC values for GL332 per organism are enumerated in Table 7.
  • MBC:MIC ratios ⁇ 4 are commonly observed for bactericidal agents and are thus considered indicative of bactericidal activity as opposed to ratios > 8 which are common among bacteriostatic agents.
  • MBC:MIC ratios for each organism is ⁇ 4, indicating that GL332 has bactericidal activity.
  • the MIC:MBC ratio for E. Coli was 1 (i.e., MIC is 0.06 mg/L and MBC is 0.06 mg/L)
  • the MIC:MBC ratio for K. pneumoniae was also 1 (i.e. , MIC of 0.03 mg/L and MBC of 0.03 mg/L)
  • the MIC:MBC ratio for P. aeruginosa PA01 was also 1 (i.e., MIC of 0.5 mg/L and MBC of 0.5 mg/L).
  • PC unspecified KPC class A serine carbapenemase.
  • ESBL extended spectrum b lactamase
  • CTX-M-14 is an extended spectrum b lactamase
  • QC quality control strain
  • Col R colistin resistant
  • GL332 has very potent bactericidal activity against each of the tested organisms E. coli, K. pneumoniae and P. aeruginosa.
  • the MIC:MBC values for E. coli, K. pneumoniae and P. aeruginosa were ⁇ 4, indicating that GL-332 exhibits strong bactericidal activity rather than the lesser bacteriostatic activity.
  • Plasma protein binding is an important consideration for drug discovery as it influences the free fraction of drug available to bind to its target.
  • Human serum albumin (HAS) is an abundant extracellular protein found in the blood plasma and tissue fluids. HAS is able to bind and then transport a number of endogenous compounds found in the body. However, HAS also absorbs a significant amount of drug in plasma and tissue fluids. The pharmacological activity of a drug is, in part, determined by the concentration of unbound concentration of the drug, not the total concentration of the blood in the plasma and tissue fluids. Accordingly, drugs that show a high affinity for HAS require higher dosing in order to achieve a therapeutic effect.
  • GL-332 was evaluated for in vitro activity against a panel of Gram-negative aerobic bacteria in the presence and absence of mouse serum to determine the effects of plasma proteins on the MIC values for GL-332.
  • GL-332 The activity of GL-332 was evaluated in the presence and absence of mouse serum by conducting MIC testing against a range of Gram-negative aerobic bacteria. Isolates were chosen based on in vitro activity of GL-332. Studies were conducted following guidelines published by the Clinical and Laboratory Standards Institute using broth microdilution; concentrations of mouse serum used during testing were 0, 20 and 50%. Imipenem, amikacin, levofloxacin, and meropenem were the comparator agents in this study.

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Abstract

The present application provides novel cephem, penem, and monobactam compounds that exhibit antibiotic activity against both Gram-negative and Gram-positive bacteria, as well as compositions comprising these compounds and methods of using these compounds and compositions to treat bacterial infections.

Description

CEPHEM COMPOUNDS WITH LATENT REACTIVE GROUPS AND METHODS OF USING AND MAKING SAME
PRIORITY CLAIM
[0001] This application claims the benefit of U.S. Provisional Appl. No. 62/828,940, filed April 3, 2019, the disclosure of which is incorporated herein in its entirety, including drawings.
BACKGROUND
[0002] Antimicrobial resistance is currently so severe that infecting pathogens are most often described in terms of their resistance to antibiotics. Categories include: usual drug resistance, multi-drug resistance, extreme drug resistance and pan-drug resistance [1 ] The need for new antibiotics which are effective against such resistance is crucial [2, 3-8]
[0003] Beta-lactam antibiotics are one of the most important and successful classes of antimicrobials for the treatment of human infections caused by bacterial pathogens [9] Bacterial resistance to beta-lactam antibiotics is typically facilitated by production of beta-lactamases, which inactivate beta-lactam antibiotics before they can bind their targets [10-12]
[0004] Cephalosporins, a sub-class of beta-lactam antibiotics, were developed in response to the appearance of early beta-lactamases (penicillinases). Thereafter, bacteria evolved beta-lactamases capable of hydrolyzing these first-generation cephalosporins [13, 14]
[0005] A wide variety of chemical modifications have been assessed to restore effectiveness of cephalosporins, including modifications at the 7-side chain [1 , 15, 16] and modifications at the 3-methyl position of the cephem ring.
[0006] U.S. Patent Nos. 8,883,772; 9,453,032; 9,809,605; 9,862,729; 9,975,905; and 10,000,509; U.S. Patent Publ. Nos. 2018/0273549 and 2018/0319817; and PCT Publ. Nos. W009/49086 and WO4/24503 relate to cephems and related compounds modified with certain latent reactive groups. The compounds disclosed in these patent documents exhibit time-dependent inhibition of beta-lactamases. Certain of these previously disclosed compounds contain a styryl group or a substituted styryl group to conjugate the latent reactive group with the cephem ring, and certain compounds have monobactam rings. Each of these documents is incorporated by reference herein in its entirety for the cephems, cephalosporins, and compounds of related beta-lactam ring structures disclosed therein, as well as the disclosed methods of synthesizing said compounds.
[0007] U.S. Patent Nos. 7,384,928; 7,696,354; 8,883,773; 9,085,589; 9, 145,425; 9,290,515; 9,238,65; and 9,334,289 disclose cephem and/or cephalosporin compounds having a catechol or pseudocatechol group. Each of these patents is incorporated by reference herein in its entirety for the cephem and cephalosporin structures disclosed therein, as well as the disclosed methods of synthesizing cephems and cephalosporins.
[0008] U.S. Patent No. 9,340,566 discloses cephalosporin antibiotics having a moiety containing a five-member ring with a positively charged N substituted on the cephem ring. This patent is incorporated by reference herein in its entirety for the disclosed cephalosporin structures and methods of synthesizing those compounds.
[0009] U.S. Patent Nos. 4,465,632; 4,689,292; 4,729,993; 4,978659; 5,334,590; 5,342,933; 5,350,746; 5,382,575; 6,677,331 ; 6,825, 187; 7,468,364; 7,632,828; 8318,716; 9, 145,425; and 9,937, 151 disclose carbapenem compounds. Each of these patents is incorporated by reference herein in its entirety for the disclosed carbapenem structures and methods of synthesizing those compounds.
[0010] U.S. Patent Nos. 4,218,459; 4,298 741 ; 5,055,463; 5,138,050; and 5,395,931 relate to carbapenem compounds having a 6-amido group and provide some detail of synthesis of such compounds. Each of these patents is incorporated by reference herein in its entirety for the disclosed carbapenem structures and methods of synthesizing those compounds, particularly 6-amido substituted carbapenems.
[0011] U.S. Patent No. 4,347,355 discloses transpeptidase inhibitors that are beta- lactams of the general structure:
Figure imgf000004_0001
[0012] where R, among others, is an acyl group, and R' is hydrogen, lower alkoxy, lower alkoxyalkyl, lower alkyl, phenylthio or lower alkylmercapto. This patent is incorporated by reference herein its entirety for the disclosed substituted carbepenem compounds therein and methods of synthesizing those compounds.
[0013] U.S. Patent Nos. 5,036,063; 5, 1 16,832; 5,703,068; and 6,271 ,222 disclose penem compounds. Each of these patents is incorporated by reference herein in its entirety for the disclosed penem structures and methods of synthesizing those compounds.
[0014] U.S. Patent No. 9, 174,978 and corresponding PCT Publ. No.
W015/148379; U.S. Patent Nos. 9,556, 165 and 9,782,390 and corresponding PCT Publ. No. 2013/1 10643; U.S. Patent Publ. No. 2012/0302542 and corresponding PCT Publ. No. W012/73138; and U.S. Patent No. 8,252,782 and 8,324,198 and corresponding PCT Publ. No. W010/70523 all disclose compounds for treating bacterial infections which contain a monobactam ring with certain acylamino groups at the 3 position of the ring. Each of these patents documents is incorporated by reference herein in its entirety for the disclosed monobactam structures and methods of synthesizing those compounds.
[0015] Despite the many structural modifications of cephalosporin and related cephem, penem and monobactam antimicrobials, there remains an urgent unmet medical need for more effective antimicrobials to treat infections caused by drug- resistant bacterial pathogens, particularly Gram-negative bacterial pathogens and especially those beta-lactam resistance bacteria which produce metallo-beta- lactamases, carbapenemases and extended-spectrum beta-lactamases. There is a particularly urgent need for antimicrobials to treat infections caused by bacteria that are resistant to carbapenems, including Enterobacteriaceae (e.g., Escherichia coli), Acinetobacter baumannii, and most particularly Pseudomonas aeruginosa [6, 17-23]
SUMMARY
[0016] Provided herein in certain embodiments are cephem, penem and monobactam compounds having certain aryl and heteroaryl groups to which a latent reactive group is bonded. The aryl and heteroaryl groups are bonded to the beta-lactam ring and the latent reactive group is conjugated to beta-lactam ring via the aryl or heteroaryl group. In certain embodiments, the latent reactive group is a leaving group having a positively charged nitrogen, and in certain of these embodiments the latent reactive group contains a vicinal diol or is bonded to an unsubstituted or substituted catechol. The compounds contain a latent reactive group which is released on hydrolysis/cleavage of the lactam ring and is believed to function to inhibit beta- lactamases. More specifically, the cephems are cephalosporins, cephamycins, carbacephems, and oxacephems. More specifically, the penems are penems (i.e. , an unsaturated b-lactam with a sulfur atom in the five-member ring), carbapenems (i.e., an unsaturated b-lactam with a carbon atom in the five-member ring) or oxapenems (i.e., an unsaturated b-lactam with an oxygen atom in the five-member ring). Preferred cephems are cephalosporins. Preferred penems are carbapenems. In specific embodiments, the compounds are monobactams. In specific embodiments, the compounds are cephalosporins. In specific embodiments, the compounds are carbapenems. In specific embodiments, the aryl groups are unsubstituted or substituted phenyl groups. In specific embodiments, the heteroaryl groups are unsubstituted or substituted single ring heteroaryl groups having one, two, three or four nitrogens in the ring.
[0017] In certain embodiments, the compounds provided herein have the structure set forth in Formula I:
Figure imgf000006_0001
Formula I where:
R is an acylamino group (R1CO-NFI-) or an alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group;
M is a divalent cephem, penem or monobactam ring selected from:
Figure imgf000007_0001
where:
each RA is independently hydrogen, a C1 -C3 alkyl group or a C1 -C3 alkoxy group;
RB is hydrogen, a C1 -C3 alkyl group or a C1 -C3-alkoxy group;
Rc is hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, acyloxy-substituted alkyl, a carboxyl protecting group or, when the CO2 group to which Rc is attached is negatively charged, a pharmaceutically acceptable cation;
Zi and Z2 are independently, S, SO, SO2, 0, or C(RE)2, where each RE is independently hydrogen or a C1 -C3 alkyl group; L is an optional linker which is a substituted or unsubstituted ethylene or a substituted or unsubstituted phenyl group and n is 0, if the linker is absent, or is an integer ranging from 1 to 5, if the linker is present; the B ring is an unsubstituted or substituted phenyl ring or an unsubstituted or substituted 6-member heteroaryl ring having one to four ring nitrogens; and
R5, R6, R7, Re and R9 are independently selected from hydrogen, halogen, cyano, nitro, C1 -C3 alkyl, C1 -C3 haloalkyl, amino, C1 -C3 alkylamino, and C1 -C3 dialkylamino and -CH2-X, with the exception that when the ring atom to which one of R5-R9 is bonded is a nitrogen, that R5-R9 is not present or is independently selected from hydrogen or C1 -C3 alkyl and at least one of Rs to R9 is -CH2-X, where X is an organic or inorganic leaving group.
[0018] Optional substitution for L groups includes one or more halogen, nitro, cyano, C1 -C3 alkyl, C1 -C3 alkoxy, or -C02Rc groups, where Rc is as defined in Formula I .
[0019] In specific embodiments, R7 is -CH2-X. In specific embodiments, the B ring is an unsubstituted phenyl or a mono-nitro-substituted phenyl and R7 is -CH2-X. In specific embodiments, the B ring is an unsubstituted 6-member heteroaryl ring having one nitrogen and R7 is -CH2-X.
[0020] In specific embodiments, X is halogen, a C1-C6 ester, a C1-C6 thioester, a C1-C6 alcohol, a C1-C6 alkylamino, a C1-C6 dialkylamino, a Ci -C6 trialkylammonium, a C1-C6 phosphate ester, a C1-C6 phosphite ester, a C1-C6 sulfate ester, a C1-C6 sulfite ester, a thiol, a sulfenyl, an unsubstituted or substituted phenoxy, an unsubstituted or substituted tosyl or a leaving group having a positively charged nitrogen, such as an unsubstituted or substituted pyridinium group. In specific embodiments, X is a leaving group having a positively charged nitrogen wherein the leaving group contains a vicinal diol or is bonded directly or indirectly to an unsubstituted or substituted catechol.
[0021] In specific embodiments, -CH2-X has structure set forth in Formula Xi :
Figure imgf000009_0001
Formula Xi
where:
A+ is a leaving group containing a positively charged nitrogen;
Rii , if present, is selected from hydrogen, hydroxyl, halogen, C1 -C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1 -C3 alkyl, unsubstituted or substituted C1 -C3 alkoxy, cyano, and nitro groups or
R11 with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; and
D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two or three heteroatoms (preferably N, S or 0), where m is 1 or 0 to indicate presence or absence of linker D. Optional substitution for the linker D includes one or more halogen, oxo group, C1 -C3 alkyl group, or C1 -C3 alkoxy group.
[0022] The A+ group in Formula Xi is bonded to the indicated phenyl ring through linker D or is a partially or fully unsaturated heterocyclic ring fused to the indicated phenyl ring.
[0023] In certain embodiments of Formula Xi, the indicated phenyl ring is a catechol ring having two hydroxyl groups substituted on any two adjacent ring carbons.
[0024] In specific embodiments of Formula I, where X is Formula Xi, two, three or four of R12-R15 are hydroxyl groups. In specific embodiments of Formula I, where X is Formula X-i, two of R12-R15 are hydroxyl groups. In specific embodiments of Formula I, where X is Formula X-i , R7 is -CH2-X1.
[0025] Beta-lactam ring systems of the compounds of Formula I include those of cephems, cephamycins, carbacephems, oxacephems, penems, carbapenems, oxapenems, or monobactams.
[0026] In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable salt of the compound of Formula I. The salt of the compound may be formed at the carboxylate of the group at the 2-position of the cephem/penem ring; and/or at a carboxylate and/or or at an amino group in the group at the 7-position (cephem) or 6-position (penem) of the ring; and/or at the positively charged nitrogen of the M group. The compound can also be zwitterionic. In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable solvate of the compound or salt of Formula I. In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable hydrate of the compound or salt of Formula I .
[0027] In specific embodiments, Zi is S, SO, or SO2. In specific embodiments, Zi is S. In specific embodiments, Zi is CH2 or CHRE, where RE is hydrogen or a C1 -C3 alkyl group. More specifically, RE is methyl.
[0028] In specific embodiments, Zi is CH2. In specific embodiments, Zi is O.
[0029] In specific embodiments, Z2 is S, SO, or SO2. In specific embodiments, Z2 is S. In specific embodiments, Z2 is CH2 or CHRE, where RE is hydrogen or a C1 -C3 alkyl group. More specifically, RE is methyl.
[0030] In specific embodiments, Z2 is CH2. In specific embodiments, Z2 is O.
[0031] In specific embodiments, R is an optionally substituted alkyl group. In specific embodiments, R is a hydroxy substituted alkyl group, a hydroxy substituted alkyl group where the hydroxyl group is protected or a halogen substituted alkyl group. More specifically, R is a hydroxy substituted C1 -C6 alkyl group, where the hydroxyl group is optionally protected. More specifically, R is a halogen substituted C1 -C6 alkyl group. More specifically, R is a fluorine substituted C1 -C6 alkyl group. Yet more specifically, R is a hydroxyl substituted C1 -C3 alkyl group, where the hydroxyl group is optionally protected. In specific embodiments, the C1 -C6 alkyl or the C1 -C3 alkyl is substituted with a hydroxy or a halogen on the 1 -position of the alkyl group. In a specific embodiment, R is 1 -hydroxyethyl, where the hydroxyl group is optionally protected. In a specific embodiment, R is a 1 -fluoroethyl. In a specific embodiment, R is 1 - hydroxyethyl.
[0032] In specific embodiments, R is an acylamino group, particularly an acylamino group of a cephalosporin, cephamycin, carbacephem or oxacephem antibiotic. In an embodiment R is R-i-CO-NH-, where Ri is any of various organic groups, including without limit, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted carbocyclic, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl. In specific embodiments, R1CONH- is an acylamino group of a known beta-lactam antibiotic. A wide variety of beta-lactam antibiotics is known in the art. Acylamino groups of representative known beta-lactam antibiotics are described hereinafter. In an embodiment, Ri is an optionally substituted benzyl group.
[0033] In specific embodiments, the compounds of Formula I are cephalosporins, where M is Mi , Zi is S and RB is hydrogen. In specific embodiments, the compounds of Formula I are cephalosporins, where M is Mi , Zi is S, R is an acylamino group and RB is hydrogen. In more specific embodiments, in the compounds of Formula I, M is Mi, Zi is S, RB is hydrogen and both of RA are hydrogen. In more specific embodiments, in the compounds of Formula I, M is Mi , Zi is S, R is an acylamino group, RB is hydrogen and both of RA are hydrogen. In specific embodiments of cephalosporins, both RA are hydrogen. In specific embodiments of cephalosporins, one RA is a C1 -C3 alkyl and the other RA is hydrogen. In specific embodiments of cephalosporins, one RA is a methyl and the other RA is hydrogen.
[0034] In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and RB is C1 -C3 alkyoxy. I In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and RB is C1 -C3 alkyoxy. In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and RB is methoxy. In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and RB is methoxy. In specific embodiments of cephamycins, both RA are hydrogen. In specific embodiments of cephamycins, one RA is a C1 -C3 alkyl and the other RA is hydrogen. In specific embodiments of cephamycins, one RA is a methyl and the other RA is hydrogen.
[0035] In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CH2 or CFKCFh). In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2, or more specifically Z2 is CFI2 or CH(CH3) and RB is hydrogen. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2, or more specifically Z2 is CFI2 or CH(CH3), RB is hydrogen and R is a C1 -C6 hydroxyalkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CFI2 or CH(CH3), RB is hydrogen and R is a 1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, R is an (R)-1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CFI2 or CH(CH3), RB is hydrogen and R is a 1 -hydroxyethyl group. In specific embodiments, R is an (R)-1 -hydroxyethyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CFI2 or CH(CH3), RB is hydrogen and R is an acylamino group.
[0036] In specific embodiments, the compounds of Formula I are monobactams where M is M3 and n is 0. In specific embodiments, the compounds of Formula I are monobactams where M is M3, L is— CH=CH-, n is 1 and the B ring is either cis or trans with respect to the nitrogen of the monobactam ring. In specific embodiments, the compounds of Formula I are monobactams where M is M3, L is -C(C02Rc)=CFI-, n is 1 and the B ring is either cis or trans with respect to the nitrogen of the monobactam ring.
[0037] In specific embodiments, compounds of Formula I have a cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have a 5- or 6- member cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have an isoquinoline diol or a quinolone diol group.
[0038] In specific embodiments, -CFI2-X has the structure set forth in Formula X2:
Figure imgf000013_0001
where:
Rii and R12- R-is are generally as defined in Formula Xi or
R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
R16 is hydrogen or unsubstituted or substituted C1 -C8 alkyl or cycloalkyl;
Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl;
R18 is a divalent -(CH2)P- moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and Rie together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds. [0039] In an embodiment, the phenyl ring at the right of the Formula X2 is substituted with at least two hydroxyl groups on adjacent ring carbons and is bonded to the positively charged N through the R18 moiety or is fused to the 5- or 6-member ring formed by R17 and R18.
[0040] In specific embodiments of X groups, optional substitution is substitution with one or more C1 -C3 alkyl group, C1 -C3 alkoxy group, hydroxyl, halogen, carboxylate or esters thereof, nitro, or cyano. Specific halogens include chlorine or fluorine.
[0041] In specific embodiments, the M group of Formula I is selected from any M group from M1 -M3. In specific embodiments, the B ring of Formula I is selected from any B ring from B-i-Bse. In specific embodiments, the M group of Formula I is selected from any M group from M1 -M3 and the B ring of Formula I is selected from any B ring from B1 -B58.
[0042] In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, or B14.
[0043] In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, or Bi4.
[0044] In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, B 14 , B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, or B14. [0045] In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi , B7, Be, or B14.
[0046] In specific embodiments, the M group of Formula I is selected from M3-23, or M3-25, and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36. specific embodiments, the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi , B7, Be, or Bi4.
[0047] In certain embodiments, the compounds provided herein exhibit antibiotic activity against Gram-negative bacteria, Gram-positive bacteria, or both Gram-negative and Gram-positive bacteria. In certain embodiments, the compounds provided herein exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In certain embodiments, the compounds provided herein exhibit antibiotic activity against bacteria which produce various beta-lactamases.
[0048] In certain embodiments, the compounds provided herein exhibit antibiotic activity against one or more Enterobactenaceae, including but not limited to Escherichia coir, Klebsiella ; Proteus, Citrobacter; Serratia; and/or Enterobacter. In certain of these embodiments, the compounds exhibit antibiotic activity against strains of Klebsiella pneumoniae ; Klebsiella oxytoca ; Proteus mirabilis ; Citrobacter freundir, Serratia marcescens ; Enterobacter aerogenes ; and/or Enterobacter cloacae.
[0049] In certain embodiments, the compounds provided herein exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL). In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemase. In some embodiments, the compounds exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL), which produce AmpC beta-lactamases or which produce a carbapenemase.
[0050] Provided herein in certain embodiments are compositions comprising one or more of the cephem, penem or monobactam compounds disclosed herein. In certain of these embodiments, the compositions are pharmaceutical compositions comprising a pharmaceutically effective amount of the one or more compounds. In these embodiments, the pharmaceutical composition comprises a sufficient amount of the one or more compounds to inhibit bacterial growth.
[0051] In certain embodiments, the pharmaceutical compositions provided herein comprise a beta-lactamase inhibitor or a beta-lactam antibiotic other than a compound of any one of the formulas herein. In certain embodiments, the pharmaceutical compositions provided herein contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitor other than a compound of Formula I, one or more beta-lactam antibiotic other than a compound of Formula I, one or more monobactam other than a compound of Formula I, one or more carbapenem other than a compound of Formula I or one or more aminoglycoside antibiotic. Pharmaceutical compositions herein optionally further comprise one or more pharmaceutically acceptable carriers or excipients as are known in the art. In certain embodiments, pharmaceutical compositions herein are for administration by injection. In certain embodiments, pharmaceutical compositions herein are for oral administration.
[0052] Provided herein in certain embodiments are methods of treating a bacterial infection in a subject, which may be a human or non-human animal, preferably a mammal, by administering one or more of the compounds or pharmaceutical compositions provided herein.
[0053] Provided herein in certain embodiments are the compounds and pharmaceutical compositions provided herein for use in the prevention or treatment of bacterial infections.
[0054] Provided herein in certain embodiments is the use of the compounds disclosed herein for the preparation of a medicament for the prevention or treatment of a bacterial infection. In certain embodiments, the bacterial infection is an infection by a bacterium that is a multi-drug resistant bacterium. In certain embodiments, the bacterial infection is an infection by a bacterium which produces extended spectrum beta- lactamases.
[0055] Other embodiments of the disclosure will be apparent to one of ordinary skill in the art on review of the Drawings and Detailed Description that follow.
BRIEF DESCRIPTION OF THE DRAWINGS
[0056] FIG. 1 is a graph depicting the mean plasma concentration of GL-332 and the comparators, ceftazidime, and ceftriaxone after IV dosing. The data shows that GL- 332 has pharmacokinetic profile similar to other clinically available cephalosporins with an early elimination rate similar to ceftazidime and a terminal elimination half-life similar to ceftriaxone.
[0057] FIG. 2 shows an ICso inhibition curve of GL-332 for PBP3. The data shows that GL-332 exhibited a very low ICso of PBP3 at a concentration of 10 ng/mL.
[0058] FIG. 3 shows the effects of GL-332 on binding to PBP3 and thereby, inhibiting bacterial growth using electrophoresis. As demonstrated in FIG. 3, as the concentration of GL-332 is increased (10 fg/mL to 100 pg/mL), the fluorescent signal decreases for both replicate 1 and replicate 2.
[0059] FIG. 4 is a graph depicting the survival rate (100% survival, p < 0.05) after also SC dosing in mice. The data shows that the survival rate for the mice administered the vehicle control rapidly decreased at day 1 ; however, the survival rate for GL-332 (QD and BID) was 100% after day 7, comparable to gentamicin.
[0060] FIGs. 5A-5G shows plots of Vo (initial rate without GL-332)AA (initial rate with GL332 of hydrolysis nitrocefin by AmpC, KPC-2, CTX-M-15, NDM-1 , IMP-1 , OXA- 24, and OXA-48, respectively and best fit lines to a competitive inhibition model. FIGs. 5A-5G reveal that GL-332 inhibited all of the b-lactamases via a competitive mechanism.
[0061] FIGs. 6A-6C are graphs depicting the plasma concentration of GL-337 after IV, SC, and mean concentration after dosing at 20 mg/kg, respectively. As demonstrated by FIGs. 6B-6C, GL-337 was rapidly absorbed at 20 mg/kg in mice after subcutaneous dosing. DETAILED DESCRIPTION
[0062] The following description of the invention is merely intended to illustrate various embodiments of the invention. As such, the specific modifications discussed are not to be construed as limitations on the scope of the invention. It will be apparent to one skilled in the art that various equivalents, changes, and modifications may be made without departing from the scope of the invention, and it is understood that such equivalent embodiments are to be included herein.
[0063] As disclosed herein, the cephem GL-332 has been found to have potent bactericidal activity against a broad range of different organisms, including E. coli, K. pneumoniae, and P. aeruginosa. GL-332 was found to inhibit bacterial growth and replication at remarkably low concentrations. The results provided herein further show that GL-332 has a low affinity for plasma proteins and inhibits penicillin-binding proteins (PBP). For example, GL-332 can inhibit bacterial growth as evidenced by the MIC data and it is contemplated that GL-332 exerts its effect predominately by potently binding to PBP3 with an ICso as low as 10 ng/mL.
[0064] The results provided herein also establish that GL-332 exhibits profound protective effects against bacterial infections in vivo. For example, intravitreal administration of GL-332 to mice infected with P. aeruginosa resulted in a 100% mice survival rate after 7 days. Kinetic studies together with ICso values indicate that GL-332 is a suitable drug candidate for the development of more potent b-lactamase inhibitors, and that it is not susceptible to inactivation by serine b-lactamases from the Ambler class enzymes A-D.
[0065] The results provided herein establish GL-332 as superior to previously disclosed antibacterial compounds. Accordingly, provided herein is GL-332 and salts or solvates thereof, as well as pharmaceutical compositions comprising GL-332 or salts or solvates thereof. Also provided herein are additional cephem, penem, and monobactam compounds sharing structural characteristics with GL-332, including salts and solvates of these compounds, as well as pharmaceutical compositions comprising these compounds. Further provided herein are methods of using the compounds and pharmaceutical compositions provided herein to inhibit bacterial growth and/or replication and to treat bacterial infection, and the use of the disclosed compounds and pharmaceutical compositions for these methods. Further provided are kits comprising the compounds and pharmaceutical compositions provided herein.
Compounds
[0066] Provided herein in certain embodiments are novel cephem, penem and monobactam compounds having (1 ) a phenyl or (2) a heteroaryl moiety, such as a pyridyl group, at the 3-position in the cephem or penem ring or at the N of the monobactam ring, to which a leaving group X is bonded through a methylene group (- CH2-X). In specific embodiments, the leaving group has a positively charged N atom. In a more specific embodiment, the leaving group also contains a vicinal diol or is bonded to an unsubstituted or substituted catechol. In some embodiments, the leaving group is a positively charged leaving group.
[0067] Provided herein in other embodiments are monobactam compounds having a styrenylmethylene moiety at the N of the monobactam ring to which a leaving group is bonded. In specific embodiments, the leaving group has a positively charged N atom. In a more specific embodiment, the leaving group also contains a vicinal diol or is bonded to an unsubstituted or substituted catechol.
[0068] The compounds provided herein contain a latent reactive group which is released on hydrolysis/cleavage of the lactam ring and is believed to function to inhibit beta-lactamases. The beta-lactams of the invention include cephems, penems and monobactams. More specifically, the cephems are cephalosporins, cephamycins, carbacephems, and oxacephems. More specifically, the penems are penems, carbapenems or oxapenems. Preferred cephems are cephalosporins. Preferred penems are carbapenems.
[0069] More specifically the disclosure relates to cephem and penem compounds substituted with a phenyl or heteroaryl ring which is bonded to a positively charged leaving group and which is in turn bonded to a vicinal diol group, or more specifically a catechol moiety. In specific embodiments, the positively charged leaving group has a positively charged nitrogen.
[0070] In certain embodiments, the compounds provided herein have the structure set forth in Formula I:
Figure imgf000020_0001
Formula I,
where:
R is an acylamino group (R1CO-NH-) or an alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group;
M is a divalent cephem, penem or monobactam ring selected from:
Figure imgf000020_0002
Figure imgf000021_0001
where: each RA is independently hydrogen, a C1 -C3 alkyl group or a C1 -C3 alkoxy group;
RB is hydrogen, a C1 -C3 alkyl group or a C1 -C3-alkoxy group;
Rc is hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, acyloxy-substituted alkyl, a carboxyl protecting group or, when the CO2 group to which Rc is attached is negatively charged, a pharmaceutically acceptable cation;
Zi and Z2 are independently, S, SO, SO2, 0, or C(RE)2, where each RE is independently hydrogen or a C1 -C3 alkyl group;
L is an optional linker which is a substituted or unsubstituted ethylene or a substituted or unsubstituted phenyl group and n is 0, if the linker is absent, or is an integer ranging from 1 to 5, if the linker is present; the B ring is an unsubstituted or substituted phenyl ring or an unsubstituted or substituted 6-member heteroaryl ring having one to four ring nitrogens; and
R5, R6, R7, Re and R9 are independently selected from hydrogen, halogen, cyano, nitro, C1 -C3 alkyl, C1 -C3 haloalkyl, amino, C1 -C3 alkylamino, and C1 -C3 dialkylamino and -CH2-X, with the exception that when the ring atom to which one of R5-R9 is bonded is a nitrogen, that R5-R9 is not present or is independently selected from hydrogen or C1 -C3 alkyl and at least one of Rs to R9 is -CH2-X, where X is an organic or inorganic leaving group.
[0071] Optional substitutions for L groups includes one or more halogen, nitro, cyano, C1 -C3 alkyl, C1 -C3 alkoxy, or -C02Rc groups, where Rc is as defined in Formula I .
[0072] In certain embodiments, R7 is -CH2-X. [0073] In certain embodiments, the B ring is an unsubstituted phenyl or a mono- nitro-substituted phenyl and R7 is -CH2-X.
[0074] In certain embodiments, the B ring is an unsubstituted 6-member heteroaryl ring having one nitrogen and R7 is -CH2-X.
[0075] In certain embodiments, X is halogen, a C1-C6 ester, a C1-C6 thioester, a C1-C6 alcohol, a C1-C6 alkylamino, a C1-C6 dialkylamino, a Ci -C6 trialkylammonium, a C1-C6 phosphate ester, a C1-C6 phosphite ester, a C1-C6 sulfate ester, a C1-C6 sulfite ester, a thiol, a sulfenyl, an unsubstituted or substituted phenoxy, an unsubstituted or substituted tosyl or a leaving group having a positively charged nitrogen, such as an unsubstituted or substituted pyridinium group. In specific embodiments, X is a leaving group having a positively charged nitrogen wherein the leaving group contains a vicinal diol or is bonded directly or indirectly to an unsubstituted or substituted catechol.
[0076] In specific embodiments, -CH2-X has the structure set forth in Formula Xi:
Figure imgf000022_0001
Formula Xi
where:
A+ is a leaving group containing a positively charged nitrogen;
Rii , if present, is selected from hydrogen, hydroxyl, halogen, C1 -C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1 -C3 alkyl, unsubstituted or substituted C1 -C3 alkoxy, cyano, and nitro groups or Rii with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; and
D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two or three heteroatoms (preferably N, S or 0), where m is 1 or 0 to indicate presence or absence of linker D. Optional substitution for the linker D includes one or more halogen, oxo group, C1 -C3 alkyl group, or C1 -C3 alkoxy group.
[0077] The A+ group in Formula Xi is bonded to the indicated phenyl ring through linker D or is a partially or fully unsaturated heterocyclic ring fused to the indicated phenyl ring.
[0078] In a specific embodiment of Formula Xi , the indicated phenyl ring is a catechol ring having two hydroxyl groups substituted on any two adjacent ring carbons.
[0079] In specific embodiments of Formula I, where X is Formula Xi , two, three or four of R12-R15 are hydroxyl groups. In specific embodiments of Formula I, where X is Formula Xi, two of R12-R15 are hydroxyl groups. In specific embodiments of Formula I, where X is Formula Xi , R7 is -CFI2-X1.
[0080] Beta-lactam ring systems of the compounds of Formula I include those of cephems, cephamycins, carbacephems, oxacephems, penems, carbapenems, oxapenems, or monobactams.
[0081] In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable salt of the compound of Formula I. The salt of the compound may be formed at the carboxylate of the group at the 2-position of the cephem/penem ring; and/or at a carboxylate and/or or at an amino group in the group at the 7-position (cephem) or 6-position (penem) of the ring; and/or at the positively charged nitrogen of the M group. The compound can also be zwitterionic. In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable solvate of the compound or salt of Formula I. In specific embodiments, the compound of the disclosure is a pharmaceutically acceptable hydrate of the compound or salt of Formula I . [0082] In specific embodiments, Zi is S, SO, or SO2. In specific embodiments, Zi is S. In specific embodiments, Zi is Chte or CHRE, where RE is hydrogen or a C1 -C3 alkyl group. More specifically, RE is methyl.
[0083] In specific embodiments, Zi is CH2. In specific embodiments, Zi is O.
[0084] In specific embodiments, Z2 is S, SO, or SO2. In specific embodiments, Z2 is S. In specific embodiments, Z2 is Ch or CHRE, where RE is hydrogen or a C1 -C3 alkyl group. More specifically, RE is methyl.
[0085] In specific embodiments, Z2 is CH2. In specific embodiments, Z2 is O.
[0086] In specific embodiments, R is an optionally substituted alkyl group. In specific embodiments, R is a hydroxy substituted alkyl group, a hydroxy substituted alkyl group where the hydroxyl group is protected or a halogen substituted alkyl group. More specifically, R is a hydroxy substituted C1 -C6 alkyl group, where the hydroxyl group is optionally protected. More specifically, R is a halogen substituted C1 -C6 alkyl group. More specifically, R is a fluorine substituted C1 -C6 alkyl group. Yet more specifically, R is a hydroxyl substituted C1 -C3 alkyl group, where the hydroxyl group is optionally protected. In specific embodiments, the C1 -C6 alkyl or the C1 -C3 alkyl is substituted with an hydroxy or a halogen on the 1 -position of the alkyl group. In a specific embodiment, R is 1 -hydroxyethyl, where the hydroxyl group is optionally protected. In a specific embodiment, R is a 1 -fluoroethyl. In a specific embodiment, R is 1 -hydroxyethyl.
[0087] In specific embodiments, R is an acylamino group, particularly an acylamino group of a cephalosporin, cephamycin, carbacephem or oxacephem antibiotic. In an embodiment R is R1-CO-NH-, where Ri is any of various organic groups, including without limit, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted carbocyclic, optionally substituted aryl, optionally substituted heterocyclic, or optionally substituted heteroaryl. In specific embodiments, R1CONH- is an acylamino group of a known beta-lactam antibiotic. A wide variety of beta-lactam antibiotics is known in the art. Acylamino groups of representative known beta-lactam antibiotics are described hereinafter. In an embodiment, Ri is an optionally substituted benzyl group. [0088] In specific embodiments, the compounds of Formula I are cephalosporins, where M is Mi , Zi is S and RB is hydrogen. In specific embodiments, the compounds of Formula I are cephalosporins, where M is Mi , Zi is S, R is an acylamino group and RB is hydrogen. In more specific embodiments, in the compounds of Formula I, M is Mi, Zi is S, RB is hydrogen and both of RA are hydrogen. In more specific embodiments, in the compounds of Formula I, M is Mi , Zi is S, R is an acylamino group, RB is hydrogen and both of RA are hydrogen. In specific embodiments of cephalosporins, both RA are hydrogen. In specific embodiments of cephalosporins, one RA is a C1 -C3 alkyl and the other RA is hydrogen. In specific embodiments of cephalosporins, one RA is a methyl and the other RA is hydrogen.
[0089] In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and RB is C1 -C3 alkyoxy. I In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and RB is C1 -C3 alkyoxy. In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi, Zi is S, and RB is methoxy. In specific embodiments, the compounds of Formula I are cephamycins, where M is Mi , Zi is S, R is an acylamino group and RB is methoxy. In specific embodiments of cephamycins, both RA are hydrogen. In specific embodiments of cephamycins, one RA is a C1 -C3 alkyl and the other RA is hydrogen. In specific embodiments of cephamycins, one RA is a methyl and the other RA is hydrogen.
[0090] In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CH2 or CH(CH3). In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2, or more specifically Z2 is CH2 or CH(CH3) and RB is hydrogen. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2, or more specifically Z2 is CH2 or CH(CH3), RB is hydrogen and R is a C1 -C6 hydroxyalkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CH2 or CH(CH3), RB is hydrogen and R is a 1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, R is an (R)-1 -hydroxy-C1 -C6 alkyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CH2 or CH(CH3), RB is hydrogen and R is a 1 -hydroxyethyl group. In specific embodiments, R is an (R)-1 -hydroxyethyl group. In specific embodiments, the compounds of Formula I are carbapenems, where M is M2 and Z2 is C(RE)2 or more specifically Z2 is CH2 or CH(CH3), RB is hydrogen and R is an acylamino group.
[0091] In specific embodiments, the compounds of Formula I are monobactams where M is M3 and n is 0. In specific embodiments, the compounds of Formula I are monobactams where M is M3, L is— CH=CH-, n is 1 and the B ring is either cis or trans with respect to the nitrogen of the monobactam ring. In specific embodiments, the compounds of Formula I are monobactams where M is M3, L is -C(C02Rc)=CFI-, n is 1 and the B ring is either cis or trans with respect to the nitrogen of the monobactam ring.
[0092] In specific embodiments, compounds of Formula I have a cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have a 5- or 6- member cyclic quaternary ammonium group. In specific embodiments, compounds of Formula I have an isoquinoline diol or a quinolone diol group.
[0093] In specific embodiments, -CFI2-X is:
Figure imgf000026_0001
where:
R11 and R12- R-is are generally as defined in Formula Xi or
R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; Ri6 is hydrogen or unsubstituted or substituted C1 -C8 alky or cycloalkyl;
Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl;
Ri8 is a divalent -(CH2)P- moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and R18 together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds.
[0094] In certain embodiments, the phenyl ring at the right of the Formula X2 is substituted with at least two hydroxyl groups on adjacent ring carbons and is bonded to the positively charged N through the R18 moiety or is fused to the 5- or 6-member ring formed by R17 and R18.
[0095] In specific embodiments of X groups, optional substitution is substitution with one or more C1 -C3 alkyl group, C1 -C3 alkoxy group, hydroxyl, halogen, carboxylate or esters thereof, nitro, or cyano. Specific halogens include chlorine or fluorine.
[0096] In certain embodiments, the M group of Formula I is selected from any Mi , M2, or M3 group set forth below:
Figure imgf000027_0001
Mi M2 M3 [0097] In certain embodiments wherein the M group of Formula I is Mi, the Mi group is selected from M1-1-M1-21 as set forth below:
Figure imgf000028_0001
Figure imgf000029_0001
Mi-19 Mi-20 Mi-21
[0098] In certain embodiments wherein the M group of Formula I is M2, the M2 group is selected from M2-1-M2-15 as set forth below:
Figure imgf000030_0001
M2-IO M2-11 M2-12
Figure imgf000031_0001
M2-13 M2-14 M2-15
[0099] In certain embodiments wherein the M group of Formula I is M3, the M3 group is selected from M3-1-M3-25 as set forth below:
Figure imgf000031_0002
M3-8 M3-9 M3-10 M3-11
Figure imgf000032_0001
Figure imgf000033_0001
[0100] In the embodiments of M set forth above, Z-i, Z2, each RA, RB, Rc, each RD, each RE, L, and n are defined as in Formula I. RM is hydrogen or a C1 -C6 alkyl group, particularly an unsubstituted C1 -C6 alkyl group. In certain of these embodiments, RM is a C1 -C3 unsubstituted alkyl group and particularly a methyl group. In certain embodiments of M set forth above, each RD is hydrogen. In certain embodiment of M set forth above, Rc is hydrogen or a pharmaceutically acceptable cation, when the CO2 group is negatively charged. In certain embodiments of M set forth above, Rc is unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, or acyloxy-substituted alkyl. In certain embodiments of M set forth above, Rc is hydrogen. In certain embodiments of the compounds provided herein, M is selected in the alternative from M1-2 to M1-10, M2-8 to M2-13, or M3-6 to M3-25. In certain embodiments, M is selected in the alternative from M3-7, M3-9, M3-13 to M3-19. In certain embodiments, M is selected in the alternative from M3-20 to M3-25. In certain embodiments, M is selected from M3-8, M3-10, M3-11 , or M3-12. In certain embodiments, M is selected from M1-7, M1-9, M2-12, M2-13, M3-10, M3-12, M3-16, or lVh-is.
[0101] In certain embodiments, the B ring of Formula I is selected from any B ring set forth below:
Figure imgf000033_0002
Figure imgf000034_0001
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
B47 B48
Figure imgf000038_0001
B55 B56
Figure imgf000039_0001
[0102] In the embodiments of the divalent B rings set forth above, A indicates optional substitution of one or more non-hydrogen substituents on the indicated ring or optional substitution of a single non-hydrogen substituent on a given ring position. Optional non-hydrogen substituents A are selected from one or more nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, and C1-C3 trialkylammonium. More preferred A are nitro, halide, methyl, methoxy, hydroxyl, methyl sulfonyl or amino. Specific A groups are nitro, chloro, and bromo. In specific embodiments of Formulas herein, the B ring is other than a B ring having a positively charged nitrogen at the ring position attached to the M moiety or the -CH2-X moiety.
[0103] In certain embodiments of the compounds provided herein, the B ring is selected from one of B1-B7, B8-B25, B26-B33, B34-B38, or B38-B58. In certain embodiments, the B ring is selected from one of Bi, B4, B7, Be, Bn, Bn, B17, B26, or B27-B38. In certain embodiments, the B ring is one of Bi, B4, or B7, where A is halogen or nitro, Be, Bn, Bn, or Bi7. In certain embodiments, the B ring is selected from Bi, B7, Be, and Bn. In certain embodiments, the B ring is selected from B39 or B40. In certain embodiments, the B ring is other than B39 or B40. In certain embodiments, the B ring contains one N ring atom. In certain embodiments, the B ring contains two N ring atoms.
[0104] In specific embodiments, the M group of Formula I is selected from any M group set forth above and the B ring of Formula I is selected from any B ring as set forth above.
[0105] In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi , B7, Be, Bi4, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M1-6, M1-7, M1-9 or M1-10 and the B ring is selected from any one of Bi, B7, Be, or B14.
[0106] In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M2-12 or M2-13 and the B ring is selected from any one of Bi, B7, Be, or Bi4.
[0107] In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, B 14 , B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, Bi4, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-10, M3-12, M2-16, M3-17, M3-18 or M3-19 and the B ring is selected from any one of Bi, B7, Be, or B14.
[0108] In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-10, or M3-12 and the B ring is selected from any one of Bi, B7, Be, or B14.
[0109] In specific embodiments, the M group of Formula I is selected from M3-23, or M3-25, and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, B32, B34, B35 or B36. In specific embodiments, the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, B14, B26, B27, B28, or B32. In specific embodiments, the M group of Formula I is selected from M3-23, or M3-25 and the B ring is selected from any one of Bi, B7, Be, or B14.
[0110] In certain embodiments, the -CFI2X group of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000041_0001
[0111] In certain embodiments, the Xs of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000042_0001
Xs-4 X5-5 X5-6
[0112] In certain embodiments, the CQ of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000042_0002
[0113] In certain embodiments, the X7 of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000043_0001
[0114] In certain embodiments, the Xe of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000043_0004
Xs-1
[0115] In certain embodiments, the X9 of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000043_0002
X9-1 X9-2
[0116] In certain embodiments, the X10 of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000043_0003
[0117] In certain embodiments, the X11 of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000044_0001
Xl1-1
[0118] In certain embodiments, the Xi2 of any compound of Formula I, II, III, IV, or V is selected from:
Figure imgf000044_0002
X12-1
[0119] R12-R15 and Ri6 are as defined in Formula Xi or X2, and each Q is independently selected from N or C-Ax. Each W is independently selected from N-RN or C(AX)2. AX represents optional substitution with one or two non-hydrogen substituents on carbon atoms in the indicated ring or optional substitution with one or two non-hydrogen substituents on a given carbon ring position. Ax is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -C02-alkyl ester, -COFI, -COCH3, phenyl, and benzyl. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, and methoxy. In specific embodiments, Ax represents no substitution. In specific embodiments, Ax represent substitution on one ring carbon with OH, halogen (particularly Cl) or methyl.
[0120] In specific embodiments, R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate, carboxylate ester, or acyl groups. In specific embodiments, R12-R15 are all hydrogens. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol). In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12- Ri5 are hydrogen, hydroxyl, methyl, or halide. In specific embodiments, any two R12- Ri5 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, or halide. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen, chloride or both hydrogen or both chloride.
[0121] In specific embodiments, R16 is a C1 -C6 alkyl or cycloalkyl group. In specific embodiments, R16 is hydrogen. In specific embodiments, R16 is a methyl group.
[0122] In certain embodiments of the compounds provided herein, the -CH2-X group is selected from: X5; CQ; XS-I ; CQ-I ; XS-2, X5-3, X5-4, X5-5, or XS-Q; CQ-2, CQ-3, CQ-4, CQ- 5, or CQ-Q; C7, Cb, CQ, CIO, Xu or C12; C7-1 ; Cb-i ; CQ-I , or C9-2; C10-1 or C10-2; C11-1 or C11-2; or C12-1 or Xi2-2, wherein in each case variables have values as defined above.
[0123] Amine cations useful in preparation of the compounds provided herein where the X group of -CH2-X is a leaving group having a positively charged nitrogen are set forth below:
Figure imgf000045_0001
Xl03 Xl04 Xl05
Figure imgf000046_0001
Xl23 Xl24 Xl25
Figure imgf000047_0001
Xl33 Xl34 Xl35
Figure imgf000048_0001
Xl43 Xl44 Xl45
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Xl73 Xl74 Xl75
Figure imgf000053_0001
[0124] In the embodiments of the amine cations set forth above, R12-R15 and R16 are as defined in Formula Xi or X2. Ax represents optional substitution with one or more non-hydrogen substituents on carbon atoms in the indicated ring or a single optional non-hydrogen substituent on a given carbon ring position, and RN is hydrogen or a C1 - C3 alkyl group.
[0125] In certain embodiments of the amine cations set forth above, Ax is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1 -C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -C02-alkyl ester, -COH, -COCH3, phenyl, and benzyl. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3. In specific embodiments, optional non-hydrogen substituents of Ax are selected from halide, hydroxyl, methyl, and methoxy. In certain embodiments, Ax represents all hydrogens. In certain embodiments, Ax represents a single non-hydrogen substituent which is selected from halide, nitro, hydroxyl, methyl or methoxy. [0126] In certain embodiments of the amine cations set forth above, R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate, carboxylate ester, or acyl groups. In specific embodiments, R12-R15 are all hydrogens. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol). In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, hydroxyl, methyl, or halide. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl (vicinal diol) and the remaining R12-R15 are hydrogen, or halide. In specific embodiments, any two R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen, chloride or both hydrogen or both chloride.
[0127] In certain embodiments of the amine cations set forth above, R16 is a C1 -
C6 alkyl or cycloalkyl group. IN specific embodiments, R16 is hydrogen. In specific embodiments, R16 is a methyl group.
[0128] In certain embodiments of the amine cations set forth above, RN is hydrogen. In other embodiments, RN is methyl.
[0129] In certain embodiments of the compounds provided herein, the -CH2-X group is selected from those where the X group is a cation selected from X100, X120, X130, Xi4o, Xi5o, X160, Xi7o, or X180; X101-X109; X121-X129; X131-X139; X141-X149; X151-X159; X16I- X169; X171-X179; X181-X189; X107-X109; X127-X129; X137-X139; X147-X149; X157-X159; X167-X169; X177-X179; or X187-X189; wherein in each case variables have values as defined for the amine cations set forth above.
[0130] Additional -CH2-X groups of Formula I are set forth below:
Figure imgf000054_0001
X200 X201 X202
Figure imgf000055_0001
Figure imgf000056_0001
[0131] In the embodiments of the -CH2-X groups set forth above, Ay represents optional substitution with one or more non-hydrogen substituents on carbon atoms in the indicated ring or a single optional non-hydrogen substituent on a given carbon ring position. In embodiments, Ay is selected from hydrogen, nitro, halide, cyano, hydroxyl, C1 -C3 methyl, C1 -C3 alkoxy, C1-C3 alkylsulfonyl, amino, C1 -C3 alkylamino, C1-C3 dialkylamino, C1-C3 trialkylammonium, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, or unsubstituted or substituted arylalkyl. In specific embodiments, optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, methoxy, methylsulfonyl, amino, carboxylate, -CC -alkyl ester, -COH, -COCH3, phenyl, and benzyl. In specific embodiments, optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, methoxy, -CO2-CI -C3 alkylester, -COH, and -COCH3. In specific embodiments, optional non-hydrogen substituents of Ay are selected from halide, hydroxyl, methyl, and methoxy. In certain embodiments, Ay represents all hydrogens. In certain embodiments, Ay represents a single non-hydrogen substituent which is selected from halide, nitro, hydroxyl, methyl or methoxy.
[0132] In certain embodiments of the -CH2-X groups set forth above, RM represents hydrogen or an optionally substituted C1 -C6 alkyl. In certain embodiments, RM is an optionally substituted C1 -C3 alkyl group. In certain embodiments, RM is unsubstituted alkyl. In certain embodiments, RM is methyl. In certain embodiments, RM is hydrogen.
[0133] In certain embodiments of the -CH2-X groups set forth above, RN represents optionally substituted C1 -C6 alkyl groups or two or three of RN together with the nitrogen to which the RN are bound forms a 4-10 member optionally substituted heterocyclic ring. Optional substitution for RN includes one or more unsubstituted C1 - C3 alkyl group, hydroxyl or oxo (=0) groups. In certain embodiments, RN is unsubstituted C1 -C6 alkyl. In certain embodiments, Rx is selected from hydrogen, optionally substituted C1 -C6 alkyl, and optionally substituted C1 -C6 acyl groups, where optional substitution includes substitution with one or more hydroxyl, amino, alkyl amino, or dialkyl amino groups. Specific Rx are alkyl amino- or dialkyl amino-substituted C1 - C6 acyl groups. In specific embodiments, monobactams of Formula I have -CH2-X as set forth above, and in certain of these embodiments n is 1 and L is ethylene.
[0134] In specific embodiments of monobactams and carbapenems herein -CH2- X is selected from: X200-X213; X200-X205; X206; X207; X208; X209; X210; X211, X212; or X213, wherein in each case variables have values as defined above.
[0135] Exemplary Ri groups of acylamino groups of the compounds provided herein are set forth below:
Figure imgf000057_0001
Rl-8 R1-9 Rl-10 R1-11
Figure imgf000058_0001
Rl-17 R1-18 R1-19 R1-20
Figure imgf000059_0001
R1-29 R1-30
Figure imgf000060_0001
Rl-40 Rl-41 Rl-42
Figure imgf000061_0001
R1-47
[0136] The Ri groups set forth above include exemplary precursor carboxylic acids R1-COOH that are typically employed for reaction with corresponding amines (see Synthesis Schemes 1 -5 and the Examples below) to introduce the R1-CO- moiety into compounds of the disclosure. Methods for such introduction are known in the art and are exemplified herein. In certain embodiments of the compounds provided herein, Ri is a group selected from those of compounds R1-4 - R1-22. In certain of these embodiments, Ri is a group selected from those of compounds R1-4 - R1-16, and in certain of these embodiments Ri is a group selected from those of compounds R1-5, Ri- 6, R1-7, R1-8, R1-9, R1-10, R1-11 , or R1-12. In certain embodiments, Ri is a group selected from those of compounds R1-11 or R1-12. In certain embodiments, Ri is a group selected from those of compounds R1-13 or R1-14. In certain embodiments, Ri is a group selected from those of compounds R1-15 or R1-16. In certain embodiments, Ri is a group selected from those of compounds R1-17 or R1-18. In certain embodiments, Ri is a group selected from those of compounds R1-19 or R1-22. In certain embodiments, Ri is a group selected from those of compounds R1-24 or R1-25. In certain embodiments, Ri is a group selected from those of compounds R1-23, or R1-26 - R1-47. In certain embodiments, Ri is a group selected from those of compounds R1-38 - R1-43.
[0137] In certain embodiments, the compounds provided herein have a structure set forth below:
Figure imgf000062_0001
Figure imgf000063_0001
1-85 1-86 1-87
Figure imgf000064_0001
Figure imgf000065_0001
1-104 1-105
Figure imgf000066_0001
1-112 1-113
Figure imgf000067_0001
Figure imgf000068_0001
1-122 1-123
[0138] In the embodiments of the compounds set forth above, R-i , Rc, X are as defined for Formula I. In certain embodiments X is a halogen, and in other embodiments X is other than a halogen. In specific embodiments, Rc is hydrogen or -CO-ORc is carboxylate. In certain embodiments, Rc is and optionally substituted phenyl group.
[0139] In certain embodiments, the compounds provided herein are any one of Formulas I-60 to 1-123 set forth above. In certain embodiments, the disclosure provides antibacterial compounds of Formulas I-60 to I-72; I-73 to I-75; I-76 to I-78; I-79 to 1-81 ;
I -82 to I -84; I-85 to I-87; I-88 to 1-91 ; I-92 or I-93; I-94 to I-96; I-97 to I-99; 1-100 to 1-102; 1-103 or 1-105; 1-106 to 1-108; 1-109 to 1-1 1 1 ; 1-1 12 to 1-1 14; 1-1 16 or 1-1 17; 1-1 18 to 1-120; or 1-121 to 1-123; wherein in each case variables have values as defined above.
[0140] The compounds set forth above can be illustrated as positively charged species (and X groups listed can be positively charged species) where no specific anion is shown. It is appreciated in the art that such compounds are prepared as salts, for example, where the anion of the salt is a pharmaceutically acceptable cation. Certain compounds provided herein carry a -CO-0-Rc group which can be a carboxylate group. In certain embodiments, these compounds are zwitterionic. The compounds of these embodiments may be in zwitterionic form or may be in the form of an ammonium cation with an appropriate anion, such as a halide (e.g. , Cl-, Br or h), an organic anion, such as sulfate, bisulfate, acetate or trifluoroacetate, or any pharmaceutically acceptable anion, such as described herein.
[0141] In specific embodiments of Formula I, R1 is of formula:
Figure imgf000069_0001
where R22 is as defined for Ri in Formula I, X is N or CRx, where Rx is hydrogen, C1 - C3 alkyl or halogen, and particularly chlorine, and Re is hydrogen or (Rp)2PO-, where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both Rp are methyl. The R1 group is in the E or Z conformation with respect to the oxime group. More specifically the R1-1 group is in the Z conformation.
[0142] In specific embodiments of Formula I, R1 is of formula:
Figure imgf000069_0002
where Ra and Rb are as defined for Ri in Formula I, X is N or CRx, where Rx is hydrogen, C1 -C3 alkyl or halogen, and particularly chlorine, and Re is hydrogen or (Rp)2PO- where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both are methyl. More specifically, Ra and Rb are hydrogen or methyl and Re is hydrogen. Yet more specifically, Ra and Rb are both hydrogen or both methyl groups and Re is hydrogen. In more specific embodiments X is CH or N.
[0143] In specific embodiments of Formula I, R1 is of formula:
Figure imgf000070_0002
R1-3
where X is N or CRx, where Rx is hydrogen, C1 -C3 alkyl or halogen, and particularly chlorine, and Re is hydrogen or (Rp)2PO-, where each Rp is independently hydrogen, or C1 -C3 alkyl or more specifically both are methyl. Rf is hydrogen, hydroxyl, C1 -C3 alkyl, C1 -C3 alkoxy, unsubstituted or substituted phenyl or unsubstituted or substituted benzyl. Rg, Rh and Ri are independently selected from hydrogen, hydroxyl, amino, alkyl amino. More specifically, one or two of Rg, Rh, Ri are hydroxyl and the others are hydrogen. More specifically, Re is hydrogen. More specifically, Rf is hydroxyl or alkoxy. More specifically, Rf is hydroxyl.
[0144] In certain embodiments, the compounds provided herein have the structure set forth in Formula II:
Figure imgf000070_0001
Formula II
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I;
each Q is independently, N or CH; and -CFI-X2 is
Figure imgf000071_0001
where:
Rii and R12- R-is are generally as defined in Formula Xi or
R11 and R12 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms; or
R12 and R13 or R13 and R14 or R14 and R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
R16 is hydrogen or unsubstituted or substituted C1 -C6 alkyl;
Ri7 is selected from hydrogen, unsubstituted or substituted C1 -C6 alkyl;
R18 is a divalent -(CH2)P- moiety, where p is 1 -6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1 -C3 alkyl; or Ri7 and Rie together with the nitrogen to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocyclic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; where dotted lines indicate optional bonds. [0145] In embodiments of Formula II, the X group is any one of the cationic groups of Xioo-Xi89. In embodiments of Formula II, the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47. In embodiments of Formula II, the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula II, the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above. In embodiments of Formula II, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula II, the Ri group is any one of R1-2, where variables are as defined above.
[0146] In certain embodiments, the compounds provided herein have the structure set forth in Formula 11-1 :
Figure imgf000072_0001
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I; and each Q is independently, N or CH.
[0147] In embodiments of Formula 11-1 , the Ri group is derived from any one of the carboxylic acid precursors provided herein. In embodiments of Formula 11-1 , the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above. In embodiments of Formula 11-1 , the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula 11-1 , the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula 11-1 , both Q are CH. In embodiments of Formula 11-1 , one Q is N and the other is CH. In embodiments of Formula 11-1 , both Q are N.
[0148] In some embodiments of Formula 11-1 , Ri is R1-11 , Rc is hydrogen, and Q is CH. [0149] In some embodiments of Formula 11-1 , R1 is R1-5, Rc is hydrogen, and one Q is CH and the other Q is N.
[0150] In certain embodiments, the compounds provided herein have the structure set forth in Formula II-2:
Figure imgf000073_0001
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I; and each Q is independently, N or CH.
[0151] In embodiments of Formula II-2, the Ri group is derived from any one of the carboxylic acid precursors provided herein. In embodiments of Formula II-2, the Ri group is any one of Ri-i , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula II-2, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula II-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula II-2, both Q are CH. In embodiments of Formula II-2, one Q is N and the other is CH. In embodiments of Formula II-2, both Q are N.
[0152] In certain embodiments, the compounds provided herein have the structure set forth in Formula II-3:
Figure imgf000073_0002
Formula 11-3 or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I; and
each Q is independently, N or CH.
[0153] In embodiments of Formula II-3, the Ri group is derived from any one of the carboxylic acid precursors provided herein. In embodiments of Formula II-3, the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above. In embodiments of Formula II-3, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula II-3, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula II-3, both Q are CH. In embodiments of Formula II-3, one Q is N and the other is CH. In embodiments of Formula II-3, both Q are N.
[0154] In some embodiments of Formula II-3, Ri is R1-11, Rc is hydrogen, and Q is CH.
[0155] In some embodiments of Formula II-3, R1 is R1-5, Rc is hydrogen, and Q is CH.
[0156] In certain embodiments, the compounds provided herein have the structure set forth in Formula III:
Figure imgf000074_0001
or salts, or solvates thereof, where:
Ri is as defined for Formula I;
X2 is as defined for Formula II; and
each Q is independently, N or CH.
[0157] In embodiments of Formula III, the X group is any one of the cationic groups of X100-X189. In embodiments of Formula III, the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47. In embodiments of Formula III, the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula III, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula III, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula III, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula III, the -CFI2-X group is any one of the -CFI2-X groups of X200-X213.
[0158] In certain embodiments, the compounds provided herein have the structure set forth in Formula III-1
Figure imgf000075_0001
or salts, or solvates thereof, where:
Ri is as defined for Formula I and
each Q is independently, N or CH.
[0159] In embodiments of Formula III-1 , the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula MI-1 , the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula MI-1 , the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula MI-1 , the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula MI-1 , both Q are CH. In embodiments of Formula MI-1 , one Q is N and the other is CH. In embodiments of Formula MI-1 , both Q are N. [0160] In certain embodiments, the compounds provided herein have the structure set forth in Formula MI-2:
Figure imgf000076_0001
or salts, or solvates thereof, where:
Ri is as defined for Formula I and
each Q is independently, N or CH.
[0161] In embodiments of Formula III-2, the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47. In embodiments of Formula MI-2, the Ri group is any one of R1-1, R1-2 or R1-3, where variables are as defined above. In embodiments of Formula MI-2, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula MI-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula MI-2, both Q are CH. In embodiments of Formula MI-2, one Q is N and the other is CH. In embodiments of Formula MI-2, both Q are N.
[0162] In certain embodiments, the compounds provided herein have the structure set forth in Formula MI-3:
Figure imgf000076_0002
Formula III-3 or salts, or solvates thereof, where:
Ri is as defined for Formula I; and
each Q is independently, N or CH.
[0163] In embodiments of Formula III-3, the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula III-3, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula MI-3, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula MI-3, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula MI-3, both Q are CH. In embodiments of Formula MI-3, one Q is N and the other is CH. In embodiments of Formula MI-3, both Q are N.
[0164] In certain embodiments, the compounds provided herein have the structure set forth in Formula IV:
Figure imgf000077_0001
Formula IV
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I;
X2 is as defined in Formula II; each Q is independently, N or CH; and the double bond is either cis or trans.
[0165] In embodiments of Formula IV, the X group is any one of the cationic groups of X100-X189. In embodiments of Formula IV, the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula IV, the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula IV, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV, the Ri group is any one of R-M , where variables are as defined above. In embodiments of Formula IV, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV, the -CFI2-X group is any one of the -CFI2-X groups of X200-X213.
[0166] In certain embodiments, the compounds provided herein have the structure set forth in Formula IV-1 :
Figure imgf000078_0001
Formula IV-1 or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I; each Q is independently, N or CH; and the double bond is either cis or trans.
[0167] In embodiments of Formula IV-1 , the Ri group is derived from any one of the carboxylic acid precursors of R-M - R1-47. In embodiments of Formula IV-1 , the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV-1 , the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula IV-1 , the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV-1 , both Q are CH. In embodiments of Formula IV-1 , one Q is N and the other is CH. In embodiments of Formula IV-1 , both Q are N.
[0168] In certain embodiments, the compounds provided herein have the structure set forth in Formula IV-2:
Figure imgf000079_0001
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I;
each Q is independently, N or CH; and the double bond is either cis or trans.
[0169] In embodiments of Formula IV-2, the Ri group is derived from any one of the carboxylic acid precursors of R- - R1-47. In embodiments of Formula IV-2, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV-2, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula IV-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV-2, both Q are CH. In embodiments of Formula IV-2, one Q is N and the other is CH. In embodiments of Formula IV-2, both Q are N.
[0170] In certain embodiments, the compounds provided herein have the structure set forth in Formula IV-3:
Figure imgf000079_0002
or salts, or solvates thereof, where:
Ri and Rc are as defined for Formula I; each Q is independently, N or CH; and the double bond is either cis or trans.
[0171] In embodiments of Formula IV-3, the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula IV-3, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula IV-3, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula IV-3, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula IV-3, both Q are CH. In embodiments of Formula IV-3, one Q is N and the other is CH. In embodiments of Formula IV-3, both Q are N.
[0172] In certain embodiments, the compounds provided herein have the structure set forth in Formula V:
Figure imgf000080_0001
or salts, or solvates thereof, where:
R and Rc are as defined for Formula I;
X2 is as defined in Formula II; and
each Q is independently, N or CH.
[0173] In embodiments of Formula V, the X group is any one of the cationic groups of X100-X189. In embodiments of Formula V, the R is R1 CON H- and the R i group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula V, the X group is any one of the cationic groups of X100-X189 and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula V, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula V, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula V, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula V, the -CH2-X group is any one of the -CH2-X groups of X200-X213. [0174] In certain embodiments, the compounds provided herein have the structure set forth in Formula V-1 :
Figure imgf000081_0001
where:
R and Rc are as defined for Formula I;
is as defined in Formula II; and
each Q is independently, N or CH.
[0175] In embodiments of Formula V-1 , the R is R1 CONFI- and R i group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula V-1 , the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula V-1 , the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula V-1 , the Ri group is any one of Ri- 2, where variables are as defined above. In embodiments of Formula V-1 , both Q are CH. In embodiments of Formula V-1 , one Q is N and the other is CH. In embodiments of Formula V-1 , both Q are N.
[0176] In certain embodiments, the compounds provided herein have the structure set forth in Formula V-2:
Figure imgf000081_0002
where: R and Rc are as defined for Formula I;
X2 is as defined in Formula II; and
each Q is independently, N or CH.
[0177] In embodiments of Formula V-2, R is R1CONFI- and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula V-2, the Ri group is any one of R1 -1 , R1 -2 or R1 -3, where variables are as defined above. In embodiments of Formula V-2, the Ri group is any one of R1-1, where variables are as defined above. In embodiments of Formula V-2, the Ri group is any one of R1-2, where variables are as defined above. In embodiments of Formula V-2, both Q are CH. In embodiments of Formula V-2, one Q is N and the other is CH. In embodiments of Formula V-2, both Q are N.
[0178] In certain embodiments, the compounds provided herein have the structure set forth in Formula V-3:
Figure imgf000082_0001
where:
R and Rc are as defined for Formula I;
X2 is as defined in Formula II; and
each Q is independently, N or CH.
[0179] In embodiments of Formula V-3, R is R1CONH- and the Ri group is derived from any one of the carboxylic acid precursors of R1-4 - R1-47. In embodiments of Formula V-3, the Ri group is any one of R1-1 , R1-2 or R1-3, where variables are as defined above. In embodiments of Formula V-3, the Ri group is any one of R1-1 , where variables are as defined above. In embodiments of Formula V-3, the Ri group is any one of Ri- 2, where variables are as defined above. In embodiments of Formula V3, both Q are CH. In embodiments of Formula V-3, one Q is N and the other is CH. In embodiments of Formula V-3, both Q are N.
[0180] In certain embodiments of the compounds provided herein where the compounds have the structure of Formula I:
Figure imgf000083_0001
Formula I
R5, R6, R9, and Re are hydrogen;
the B ring of Formula I is:
Figure imgf000083_0002
where any two R12-R15 on adjacent ring carbons are both hydroxyl and the remaining R12-R15 are hydrogen and Ax is hydrogen;
M is:
Figure imgf000084_0001
where Rc is hydrogen; and
R is Ri, wherein Ri is
Figure imgf000084_0002
R1-11.
[0181] In certain embodiments of Formulas ll-IV, 11-1 to IV-1, II-2 to IV-2, and II-3 to IV-3, Ri is selected from the group consisting of:
R1-1;
R1-2;
R1-3;
R1-1, where R22 is a C1-C6 alkyl group;
R1-1, where R22 is methyl;
R1-1, where X is CH;
R1-1, where X is N;
R1-1, where X is CH and R22 is a 1-6 carbon alkyl group;
R1-1, where X is N and R22 is a 1-6 carbon alkyl group;
R1-1, where X is CH and R22 is methyl; R1-1 , where X is N and R22 is methyl;
R1-1 , where X is CH, R22 is methyl and R1-1 is in the Z configuration;
R1-1 , where X is N, R22 is methyl and R1-1 is in the Z configuration;
R1-2, where Ra and Rb are both C1 -C3 alkyl groups;
R1-2, where Ra and Rb are both methyl groups;
R1-2, where X is CH;
R1-2, where X is N;
R1-2, where X is CH and Ra and Rb are both methyl groups;
R1-2, where X is N and Ra and Rb are both methyl groups; and benzyl.
[0182] In certain embodiments of Formulas V and V-1 , V-2 and V-3, R is selected from the group consisting of:
R1CO-NH-;
R1CO-NH-, where Ri is R1-1 ;
R1CO-NH-, where Ri is R1-2;
R1CO-NH-, where Ri is R1-3;
R1CO-NH-, where Ri is R1-1 and R22 is a methyl group;
R1CO-NH-, where Ri is R1-1 , X is CH and R22 is a methyl group;
R1CO-NH-, where Ri is R1-1 , X is CH, R22 is a methyl group and the R1-1 is in the Z conformation;
R1CO-NH-, where Ri is R1-1 , X is N and R22 is a methyl group;
R1CO-NH-, where Ri is R1-1 , X is N, R22 is a methyl group and the R1-1 is in the Z conformation;
R1CO-NH-, where Ri is R1-2 and Ra and Rb are both methyl groups;
R1CO-NH-, where Ri is R1-2 and X is CH;
R1CO-NH-, where Ri is R1-2 and X is N;
R1CO-NH-, where Ri is R1-2 and X is CH and Ra and Rb are both methyl groups; R-iCO-NH-, where Ri is R1-2 and X is N and Ra and Rb are both methyl groups; a C1 -C6 alkyl group optionally substituted with one or more groups selected from a halogen, a hydroxy group or a protected hydroxyl group; a C1 -C6 alkyl group substituted with a hydroxy group or a protected hydroxyl group; a C1 -C6 alkyl group substituted with a hydroxy group; a 1 -hydroxyethyl group; and
Figure imgf000086_0001
[0183] In certain embodiments of Formulas ll-V, 11-1 to V-1 , II-2 to V-2 and II-3 to V-3: both Q are CH; one Q is CH and the other is N; or both Q are N.
[0184] In more specific embodiments of each of Formulas II, III, V, 11-1 , II-2, II-3, III-1 , III-2, MI-3, V-1 , V-2 and V-3, Rc is selected from the group consisting of: hydrogen;
C1 -C6 alkyl; an optionally substituted phenyl; and an optionally substituted benzyl.
[0185] In some embodiments, exemplary compounds of the disclosure which exhibit antibacterial activity as described herein are selected from Compound 504, 501 ,
505, 512, 482, 487, 502, GL357, 503, 506, 507, 513, 484, 488, 489, 476, 477, 481 , 478, 479, 490, 491 , 495, 600 (490-cis), 491 -cis, and 495-cis. The compounds are illustrated as positively charged species and it is appreciated in the art that such compounds are prepared as salts, for example, where the anion of the salt is a pharmaceutically acceptable cation. The compounds 504, 501 , 505, 512, 482, 487, 502, GL357, 503,
506, 507, 513, 484, 488, 489, 476, 477, 481 , 478, 479, 490, 491 , 495, 600 (490-cis), 491 -cis, and 495-cisare shown with a positively charged nitrogen and no specific salt is illustrated. These compounds may be in zwitterionic form or may be in the form of a ammonium cation with an appropriate anion, such as a halide (e.g., Cl , Br or I ), an organic anion, such as sulfate, bisulfate, acetate or trifluoroacetate, or any pharmaceutically acceptable anion, such as described herein.
[0186] Exemplary compounds provided herein include:
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0001
Figure imgf000091_0001
cis)
cis
[0187] In some embodiments, the compounds disclosed herein are useful for the synthesis of other compounds of Formula I. More specifically, compounds herein wherein X is Cl, Br, I, or an activated ester and wherein other potentially labile groups are protected with appropriate protecting groups can be used in syntheses described herein to prepare other compounds of Formula I.
[0188] Certain compounds of the disclosure may contain one or more stereogenic centers resulting in possible stereoisomers. It will be understood that for any such compound of the disclosure that all individual stereoisomer, any diastereomers and all mixtures thereof are included in the compounds as claimed. For any such compound the compound may be a racemic mixture of stereoisomers or may be a mixture which contains an excess of one stereoisomer and is thus optically active.
[0189] Mixtures of E and Z isomers of compounds of the disclosure may contain approximately equal amounts of E and Z isomers or may contain an excess of the E or Z an excess of the Z isomer. The amounts of E and Z isomers in a given mixture can be readily determined by standard analytical techniques, such as NMR (nuclear magnetic resonance) methods. A compound that is a given isomer (E or Z) that is free of the other isomer contains less than 2% by weight of the other isomer. A compound that is given isomer (E or Z) that is free of the other isomer preferably contains less than 1 % by weight of the other isomer. A compound that is a given isomer (E or Z) that is free of the other isomer more preferably contains less than 0.5% by weight of the other isomer. It will be appreciated in the art that when it is desired to employ a compound or intermediate that is a given E or Z isomer that it can be also desirable to minimize the amount of the other isomer present. It will however also be understood that small, albeit detectible levels of the other isomer that may be present may not be detrimental to the utility of the desired isomer.
[0190] In certain compounds of the disclosure, two variables substituted on adjacent carbons in an alkyl group or on a ring can optionally together with the atoms (typically carbons or nitrogens) to which they are bonded form a carbocyclic or heterocyclic ring. In embodiments, the ring can be a 5-member ring or a 6-member ring and may contain 0, 1 2 or 3 heteroatoms. Preferred heteroatoms for such rings are nitrogen, oxygen and sulfur. Such rings are optionally substituted, for example, with one or more halogens, hydroxyl, C1 -C3 alkyl, or carboxylate or esters thereof, or contain an oxo group (-CO-) in the ring. In an embodiment, such rings are unsubstituted. Such rings may be unsaturated (i.e., contain no double or triple bonds), partially unsaturated, i.e. , contain one or two double bonds or may be aromatic (as understood in the art).
[0191] Groups herein are optionally substituted. Most generally alky, alkenyl, and aryl, heteroaryl, and heterocyclyl groups are optionally substituted, for example, with one or more oxo group, thioxo group, halogen, nitro, cyano, cyanate, azido, thiocyano, isocyano, isothiocyano, sulfhydryl, hydroxyl, alkyl, alkoxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, aryl, aryloxy, amino (-NH2), heteroaryl, heteroaryloxy, carbocyclyl, carbocyclyloxy, heterocyclyl, heterocyclyloxy, alkylthio, alkenylthio, alkynylthio, arylthio, thioheteroaryl, thioheteroaryl, thiocarbocyclyl, thioheterocyclyl, -COR', -COH, -OCOR', -OCOH, -CO-OR', -CO-OH, -CO-O-CO-R', -CON(R')2, -CONHR', -CONH2, -NR-COR', - NHCOR', -NHR', -N(R')2, -O-SO2-R', -SO2-R', -SO2-NHR', -S02-N(R")2, -NR-SO2-R', -NH-SO2-R',- NR'CO-N(R')2, -NH-CO-NHR', -0-P0(0R')2, -0-P0(0R')(N(R')2), -0- PO(N(R')2)2, where each R' independently is an organic group and more specifically is an alkyl, cyclolkyl, alkenyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl group or two R' within the same substituent can together form a carbocyclic (containing only carbon ring members) or heterocyclic ring having 3 to 10 ring atoms. Organic groups of non hydrogen substituents are in turn optionally substituted with one or more halogens, nitro, cyano, isocyano, isothiocyano, hydroxyl, sulfhydryl, haloalkyl, hydroxyalkyl, amino, alkylamino, dialkylamino, arylalkyl, unsubstituted alkyl, unsubstituted alkenyl, unsubstituted alkynyl alkylalkenyl, alkylalkynyl, haloaryl, hydroxylaryl, alkylaryl, unsubstituted aryl, unsubstituted carbocyclic, halo-substituted carbocyclic, hydroxyl- substituted carbocyclic, alkyl-substituted carbocyclic, unsubstituted heterocyclic, unsubstituted heteroaryl, alkyl-substituted heteroaryl, or alkyl substituted heterocyclic. In specific embodiments, organic groups of non-hydrogen substituents are not further substituted. In specific embodiments, R' groups of substituents are independently selected from alkyl groups, haloalkyl groups, phenyl groups, benzyl groups and halo- substituted phenyl and benzyl groups. In specific embodiments, non-hydrogen substituents have 1 -10 carbon atoms, 1 -7 carbon atoms, 1 -5 carbon atoms or 1 -3 carbon atoms. In specific embodiments, non-hydrogen substituents have 1 -10 heteroatoms, 1 -6 heteroatoms, 1 -4 heteroatoms, or 1 , 2, or 3 heteroatoms. Heteroatoms preferably are O, N or S. In specific embodiments, substituent groups include heterocyclic groups having a single 5- or 6-member ring containing 1 , 2or 3 heteroatoms selected from N, O or S and in which one or more ring carbons or nitrogens are optionally substituted with a C1 -C3 alkyl group.
[0192] In specific embodiments, optional substitution is substitution with 1 , 2, 3, 4 or 5 non-hydrogen substituents. In specific embodiments, optional substitution is substitution with 1 or 2 non hydrogen substituents. In specific embodiments, optional substitution is substitution with 1 non hydrogen substituents. In specific embodiments, optional substitution is substitution by one or more halogen, hydroxyl group, cyano group, nitro group, oxo group, thioxo group, unsubstituted C1 -C6 alkyl group or unsubstituted aryl group. The term oxo group refers to substitution of a carbon atom with a =0, for example, to form a -CO- (carbonyl)] group.
[0193] In specific embodiments, optional substitution of phenyl rings, includes substitution with one or more halogen, one or more hydroxyl, one or more C1 -C3 alkyl, one or more C1 -C3 alkoxy, one or more nitro, or one or more cyano. In specific embodiments, optional substitution of phenyl rings, includes substitution with one or more halogen, one or more hydroxyl, one or more C1 -C3 alkyl, or one or more C1 -C3 alkoxy. In specific embodiments, optional substitution of phenyl rings, includes substitution with one or more halogen, one or more hydroxyl, or one or more C1 -C3 alkyl. In specific embodiments, optional substitution of phenyl rings, includes substitution with one halogen, one hydroxyl, one C1 -C3 alkyl, one C1 -C3 alkoxy, one nitro, and/or one cyano.
[0194] As to any of the above groups which contain one or more substituents, it is understood, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible. In addition, the compounds of this disclosure include all stereochemical isomers arising from the substitution of these compounds.
[0195] Compounds of the disclosure may contain chemical groups (acidic or basic groups) that can be in the form of salts. Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, for example, trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides (formed with hydrogen bromide), hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates (formed with maleic acid), methanesulfonates (formed with methanesulfonic acid), 2- naphthalenesulfonates, nicotinates, nitrates, oxalates, pectinates, persulfates, 3- phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), bisulfate, sulfonates (such as those mentioned herein), tartrates, thiocyanates, toluene sulfonates such as tosylates, undecanoates, and the like.
[0196] Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as benzathines, dicyclohexylamines, hydrabamines [formed with N,N-bis(dehydro- abietyl)ethylenediamine], N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen- containing groups may be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.
[0197] Compounds of the disclosure may be in the zwitterionic form.
[0198] Salts of the disclosure include "pharmaceutically acceptable salts" which refers to those salts which retain the biological effectiveness and properties of the free bases or free acids, and which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts comprise pharmaceutically-acceptable anions and/or cations.
[0199] Compounds of the disclosure can be administered in the form of pharmaceutically acceptable salts which include the following non-limiting examples: alkali metal salts, such as those of lithium, potassium and sodium; alkali earth metal salts, such as those of barium, calcium and magnesium; transition metal salts, such as those of zinc; and other metal salts, such as those of aluminum, sodium hydrogen phosphate and disodium phosphate; salts of nitrates, borates, methane sulfonates, benzene sulfonates, toluene sulfonates, salts of mineral acids, such as those of hydrochlorides, hydrobromides, hydroiodides and sulfates; salts of organic acids, such as those of acetates, trifuoroacetates, maleates, oxalates, lactates, malates, tartrates, citrates, benzoates, salicylates, ascorbates, succinates, butyrates, valerates and fumarates, amine salts, such as those of N,N'-dibenzylethylenediamine, chloroprocaine, choline, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N- methylglucamine, procaine, N-benzylphenethylamine, l-para-chlorobenzyl-2-pyrrolidin- T-ylmethyl-benzimidazole, diethylamine and other alkylamines, piperazine and tris(hydroxymethyl)aminomethane.
[0200] In specific embodiments, compound of any formula herein are salts wherein the anion of the salt is selected from halide, more specifically chloride or bromide, acetate, tosylate, tartrate, sulfate, bisulfate, succinate, phosphate, oxalate, nitrate, mesylate, maleate, malate, and citrate. In more specific embodiments, compounds of any formula herein are salts of chloride, bromide, sulfate, bisulfate, or acetate.
[0201] In specific embodiments, compounds of the disclosure are hydrates of the compounds herein and more specifically are hydrates of salts of the compounds herein. [0202] Pharmaceutically acceptable salts of the compounds of the disclosure can be derived from inorganic or organic acids or can be derived from inorganic or organic bases as is known in the art. Basic amino acids useful for salt formation include arginine, lysine and ornithine. Acidic amino acids useful for salt formation include aspartic acid and glutamic acid.
[0203] Compound of the disclosure can be administered in the form of pharmaceutically acceptable esters which include, among others, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids and boronic acids.
[0204] Those of ordinary skill in the art will appreciate that many organic compounds, including salts, can form complexes with solvents in which they are reacted or from which they are precipitated or crystalized. These complexes are known as "solvates". For example, a complex with water is known as a "hydrate". Solvates, and more particularly hydrates, of the cephem compounds of the disclosure are within the scope of the disclosure. Pharmaceutically acceptable solvates and hydrates are complexes of a compound of the disclosure with one or more solvent or water molecules, or 1 to about 100, or 1 to about 10, or one to about 2, 3 or 4, solvent or water molecules.
[0205] Certain groups in the compounds of the disclosure are optionally protected, for example, hydroxyl, carboxylate and amine groups. Methods of protection of such groups is known in the art and employed generally during synthesis again as is known in the art. T.W. Green et al. (1999) Protecting Groups in Organic Synthesis (Wiley Interscience) provides an overview of the types of protecting groups and their use. This reference is incorporated by reference herein for descriptions of types of protecting groups and methods for protecting and deprotecting. For example, compounds of the disclosure may contain a protected hydroxyl group or a protected carboxyl group. Exemplary hydroxyl protecting groups include alkoxyalkyl groups (e.g., methoxym ethyl, forming an ether at the hydroxy), C1 -C3 alkyl or C2-C6 alkenyl or arylalkyl or silyl groups (again forming ethers), or -CO-alkyl or -CO-arylalkyl (forming esters. Specific hydroxyl protecting groups include, among others, -COCFI3, -CO-t-butyl, -CO-phenyl (optionally substituted at the phenyl ring), -CO-benzyl (optionally substituted at the phenyl ring), optionally substituted benzyl, allyl, silyl (trialkylsilyl, diarylalkysilyl, aryldialkylsilyl, e.g., trimethylsily, diphenylmethylsilyl, phenyldimethylsilyl, and the like). Hydroxy groups may also be protected by formation of acetals. Carboxylate groups may be protected as esters.
Tautomers of the Compounds of Formula I
[0206] In certain embodiments, the compounds provided herein are in their tautomeric forms. For example, in some embodiments the compounds of Formula I are one of two tautomeric forms. In some embodiments, the tautomeric form of the compound influences the elimination rate and/or half-life of the compound. For example, one tautomeric form can have decreased elimination rates and/or increased drug half- life as compared to the other tautomeric form.
[0207] Exemplary tautomeric forms of the compounds are provided below for GL332, GL337, and GL357. Flowever, it will be readily apparent to one of skill in the art that any compound of Formula I can exist in either of the two tautomeric forms.
Figure imgf000098_0001
GL337 Tautomer 1 : FW = 708.7408514 GL337 Tautomer 2: FW = 707.73346 Molecular Formula: C31 H30N7O9S2 Molecular Formula: Cs-j^gNyOgSa
Figure imgf000099_0001
GL357 Tautomer 1 : FW = 709.72891 14 GL357 Tautomer 2 FW = 708.72152 Moiecuiar Formula: C30H29N8O9S2 Molecular Formula: C30H28N8Q9S2
Methods
[0208] In certain embodiments, methods are provided for treating a bacterial infection in a subject by administering one or more compounds or pharmaceutical compositions provided herein. In certain of these embodiments, the subject is a human or non-human animal, in particular a mammal.
[0209] In certain embodiments of the methods provided herein, GL-332, GL-337, GL-357, or GL-352 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof is administered to a subject to treat a bacterial infection. In some embodiments, the subject is administered GL-332 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof. In some embodiments, the subject is administered GL-357 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof. In some embodiments, the subject is administered GL-337 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof. In yet another embodiment, the subject is administered GL-352 stereoisomer, or a pharmaceutically acceptable salt, solvate, or tautomer thereof.
[0210] The disclosure relates, in particular, to cephem and penem compounds which exhibit strong antibacterial activity against certain multi-drug resistant bacteria. In specific embodiments, compounds herein exhibit low pgram/mL Minimum Inhibitory Concentrations (MIC) against bacteria which produce extended spectrum beta- lactamases.
[0211] In specific embodiments, compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one cephalosporin antibiotic.
[0212] In specific embodiments, compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one cephem or penem antibiotic.
[0213] In specific embodiments, compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which are resistant to at least one carbapenem.
[0214] In specific embodiments, compounds herein exhibit MICs of less than 100 pg/mL, of less than 50 pg/mL, less than 30 pg/mL, less than 25 pg/mL, less than 10 pg/mL, less than 5 pg/mL, less than 1 pg/mL or less than 0.5 pg/mL against bacteria which produce at least one ESBL.
[0215] In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-positive bacteria. In specific embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which produce various beta-lactamases. In embodiments, compounds of the disclosure, in particular, exhibit antibiotic activity against beta-lactamase producing Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against a wide range of Pseudomonas aeruginosa strains, including clinical isolates.
[0216] In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against Gram-positive bacteria. In specific embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which exhibit multi-drug resistance. In embodiments, compounds of the disclosure exhibit antibiotic activity against bacteria which produce various beta-lactamases. In embodiments, compounds of the disclosure in particular exhibit antibiotic activity against beta-lactamase producing Gram-negative bacteria. In embodiments, compounds of the disclosure exhibit antibiotic activity against a wide range of Pseudomonas aeruginosa strains, including clinical isolates.
[0217] In certain embodiments, compounds of the disclosure exhibit antibiotic activity against Enterobacteriaceae, including but not limited to strains of Escherichia coli ; Klebsiella ; Proteus, Citrobacter, Serratia ; and/or Enterobacter. In certain of these embodiments, the compounds provided herein exhibit antibiotic activity against strains of Klebsiella pneumoniae ; Klebsiella oxytoca ; Proteus mirabilis ; Citrobacter freundii ; Serratia marcescens ; Enterobacter aerogenes ; and/or Enterobacter cloacae.
[0218] In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce extended spectrum beta-lactamases (ESBL). In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce ESBL selected from TEM, SHV, and CTX-M. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce ESBL selected from TEM-26, SHV-1 , CTX-M-14, and CTX-M-15. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases [24] In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce AmpC beta-lactamases selected from DHA enzymes, particularly DHA-1 . In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemases. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemases selected from KPC, VIM, IMP, NDM, and OXA. In additional embodiments, compounds of the disclosure exhibit antibiotic activity against bacterial strains which produce a carbapenemase selected from KPC- 2, KPC-3, VIM-1 , IMP-1 , NDM-1 , OXA-48 and OXA-58.
[0219] The disclosure provides methods for treating or preventing bacterial infection and the symptoms and disorders associated therewith. The disclosure provides pharmaceutical compositions comprising a pharmaceutically effective amount of one or more compounds and/or salts and/or solvates of the disclosure and a pharmaceutically acceptable carrier or excipient. The compounds of the disclosure are of particular use in the treatment of infections of bacteria which exhibit resistance to or more beta-lactam antibiotics, particularly those that exhibit resistance to one or more cephalosporins and particularly those that exhibit resistance to a carbapenem. Compounds of the disclosure are of particular use in the treatment of infections of bacteria which generate one or more ESBL. Compounds of the disclosure exhibit resistance to one or more ESBL. Compounds of the disclosure are of particular use in the treatment of infections of bacteria which generate one or more ESBL. Compounds of the disclosure are of particular use in the treatment of infections of Gram-negative bacteria. Compounds of the disclosure are of particular use in the treatment of infections of Pseudomonas strains and particularly strains of Pseudomonas aeruginosa.
[0220] The compounds, salts and solvates thereof of the disclosure can be used to prepare medicaments for the treatment and prevention of bacterial infection and the symptoms and disorders associated therewith. The compounds, salts and solvates thereof of the disclosure can be used in particular to prepare medicaments for the treatment and prevention of bacterial infection of bacteria which produce one or more ESBL and the symptoms and disorders associated therewith.
[0221] To treat bacterial infections and complications thereof, the antibacterial compounds of this disclosure may be co-administered in combination with one or more antibacterial compounds, including antibacterial compounds which are cephems or penems other than those of Formulas l-lll herein and including antibacterial compounds which are not beta-lactam antibiotics. In addition, the antibacterial compounds of this disclosure may be co-administered in combination with one or more beta-lactamase inhibitors other than the compounds of Formula I herein. Co-administration includes among others separate administration of active ingredients at about the same time, combined administration in the same formulation, as well as sequential separate administration.
[0222] Compounds of the disclosure may be co-administered with a beta- lactamase inhibitor. In a specific embodiment, compounds of the disclosure may be co formulated with a beta-lactamase inhibitor. A number of such inhibitors are known in the art and one of ordinary skill in the art understands how to co-formulate such inhibitors with a given beta-lactam antibiotic. Beta-Lactamase inhibitors include among others avibactam, tazobactam, sulbactam, clavulanic acid, and relebactam. [0223] Compounds of the disclosure may be co-administered with a monobactam, such as aztreonam. Compounds of the disclosure may be co-administered with a carbapenem, other than a compound of Formula I, such as meropenem.
[0224] Compounds of the disclosure may be co-administered with a non-beta lactam antibiotic, such as an aminoglycoside antibiotic. Compounds of the disclosure may be co-administered with an aminoglycoside antibiotic, such as amikacin, gentamicin, kanamycin, neomycin, or tobramycin.
[0225] The disclosure also relates to the use of one or more compound, salt or solvate of Formula I and any sub formula thereof for the treatment of a bacterial infection or a complication of a bacterial infection. The disclosure further relates to the use of one or more pharmaceutical compositions comprising a compound, salt or solvate of Formula I and any sub formula thereof for the treatment of a bacterial infection or a complication of a bacterial infection.
[0226] The disclosure also relates to the use of one or more compound, salt or solvate of Formula I and any subformula thereof for the preparation of a medicament for the treatment of a bacterial infection or a complication thereof. The disclosure further relates to the use of one or more pharmaceutical compositions comprising a compound, salt or solvate of Formula I and any sub formula thereof for the preparation of a medicament for the treatment of a bacterial infection or a complication thereof.
Pharmaceutical Compositions
[0227] Provided herein in certain embodiments are pharmaceutical compositions comprising one or more of the compounds provided herein. Pharmaceutical compositions herein comprise a therapeutically effective amount of a cephem compound of this disclosure.
[0228] In certain embodiments the pharmaceutical compositions provided herein, contain one or more of GL-332, GL-337, GL-357, or GL-352 stereoisomer, pharmaceutical acceptable salt, or tautomer.
[0229] Pharmaceutical compositions herein comprise a therapeutically effective amount of a cephem or penem compound of any chemical formula herein. In embodiments, pharmaceutical compositions herein comprise a therapeutically effective amount of a cephalosporin of any chemical formula herein. In embodiments, pharmaceutical compositions herein comprise a therapeutically effective amount of a cephalomycin of any chemical formula herein. In embodiments, pharmaceutical compositions herein comprise a therapeutically effective amount of a carbapenem of any chemical formula herein. In embodiments, pharmaceutical compositions herein comprise a combined therapeutically effective amount of two or more cephem or penem compounds of this disclosure. In embodiments, pharmaceutical compositions herein comprise a combined therapeutically effective amount of two or more cephem or penem compounds of any formula herein and in particular any compound of Formula I.
[0230] In embodiments, pharmaceutical compositions herein optionally contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitors, other than a compound of Formula I. In embodiments, pharmaceutical compositions herein optionally contain other appropriate active ingredients including, for example, one or more beta-lactamase inhibitors, other than a cephem. In embodiments, pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more beta-lactamase antibiotics other than a compound of Formula I. In embodiments, pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more monobactam. In embodiments, pharmaceutical compositions herein contain other appropriate active ingredients including, for example, one or more carbapenem other than a compound of Formula I. In embodiments, pharmaceutical compositions herein contain other appropriate active ingredients including, for example, aminoglycoside antibiotics. Pharmaceutical compositions herein optionally further comprise one or more pharmaceutically acceptable carriers or excipients as are known in the art.
[0231] Compounds and salts of the disclosure in the form of pharmaceutical compositions or dosage forms can be administered by any known route that is appropriate for the patient being treated and for the treatment or prophylaxis that is desired. Specifically, administration can be orally or non-orally in the form of, for example, granules, powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixir, suspensions or solutions, by mixing these effective components, individually or simultaneously, with pharmaceutically acceptable carriers, excipients, binders, diluents or the like. [0232] A solid formulation for oral administration can comprise one or more of the compounds or salts, or solvates thereof, of the disclosure alone or in appropriate combination with other active ingredients. Solid formulations can be in the form of powders, granules, tablets, pills and capsules. In these cases, the instant compounds can be mixed with at least one additive, for example, sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi synthetic polymers or glycerides. These formulations can contain, as in conventional cases, further additives, for example, an inactive diluent, a lubricant such as magnesium stearate, a preservative such as paraben or sorbic acid, an anti-oxidant such as ascorbic acid, tocopherol or cysteine, a disintegrator, a binder, a thickening agent, a buffer, a sweetener, flavoring agent and/or a perfuming agent. Tablets and pills can also be prepared with enteric coating. Standard methods of formulation can be applied to preparation of formulations of the compounds and salts of the disclosure.
[0233] Non-oral administration includes subcutaneous injection, intravenous injection, intramuscular injections, intraperitoneal injection or instillation. Injectable preparations, for example, sterile injectable aqueous suspensions or oil suspensions can be prepared by known methods.
[0234] The instant pharmaceutical compositions may be formulated as known in the art for administration by inhalation, such as in the form of a nasal aerosol or nasal spray, a dry powder spray or as a solution or suspension for inhalation and may be prepared as solutions or suspensions in saline or other suitable carrier, and benzyl alcohol or other suitable preservatives, absorption promoters, fluorocarbons, or solubilizing or dispersing agents.
[0235] Rectal suppositories can be prepared by mixing the drug with a suitable vehicle, for example, cocoa butter and polyethylene glycol, which is in the solid state at ordinary temperatures, in the liquid state at temperatures in intestinal tubes and melts to release the drug.
[0236] Examples of liquid preparations for oral administration include pharmaceutically acceptable emulsions, syrups, elixirs, suspensions and solutions, which may contain an inactive diluent, for example, pharmaceutically acceptable water. [0237] The pharmaceutical composition herein can be formulated for topical administration with a suitable ointment containing one or more of the compounds or salts of the disclosure suspended or dissolved in a carrier, which include mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and pharmaceutically acceptable water. In addition, topical formulations can be formulated with a lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and pharmaceutically acceptable water.
[0238] As is understood in the art, dosages of therapeutic compounds are dependent on age, body weight, general health conditions, sex, diet, dose interval, administration routes, excretion rate, combinations of drugs and conditions of the diseases treated. While taking these and other necessary factors into consideration, generally, dosage levels of between about 10 pg per day to about 5000 mg per day, preferably between about 100 mg per day to about 1000 mg per day of the compound are useful in the treatment of bacterial infection. Typically, the pharmaceutical compositions of this disclosure will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier or excipient materials to produce a single dosage form will vary depending upon the patient/individual treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (W/W). Preferably, such preparations contain from about 20% to about 80% active compound.
[0239] While these dosage ranges can be adjusted by a necessary unit base for dividing a daily dose, as described above, such doses are decided depending on the diseases to be treated, conditions of such diseases, the age, body weight, general health conditions, sex, diet of the patient then treated, dose intervals, administration routes, excretion rate, and combinations of drugs. While taking these and other necessary factors into consideration., for example, a typical preparation will contain from about 0.05% to about 95% active compound (W/W). Preferably, such preparations contain from about 10% to about 80% active compound. The unit dose of the composition of this disclosure is administered once or multiple times daily. [0240] A desirable dosage form for antibiotic compounds of the disclosure is an oral dosage form to be administered once a day or twice a day.
[0241] Another desirable dosage form for antibiotic compounds of the disclosure is an parenteral dosage form to be administered once a day or twice a day.
Synthesis of Compounds of Formula I
[0242] Exemplary synthetic schemes for preparation of the compounds provided herein are provided in Schemes 1 -5.
[0243] US Patent Nos. 4,840,945; 5,036,034; and 7,129,232; and US Patent Publ. No. 2005/0043531 provide additional methods of synthesis of cephalosporin compounds, and in particular provide examples of the synthesis of useful acylamino groups. Each of these references is incorporated by reference herein in its entirety for such descriptions.
[0244] It will be apparent to one of ordinary skill in the art on review of the synthetic schemes of Schemes 1 -5 that additional compounds of Formula I can be prepared by substitution of certain starting materials, e.g., amino salts or carboxylic acids for introduction of acylamino groups. Additionally, various B rings can be introduced to form additional compounds of the disclosure as illustrated and exemplified in these Schemes again by choice of starting materials. Appendix A provides additional details of synthetic methods useful in the preparation of compounds herein. In particular, US Patent Appl. No. 16/151 ,479, filed October 4, 2018, incorporated herein by reference, provides details of the synthesis of Amine salts that are useful in the preparation of compounds herein of Formula I. In some embodiments, any of the exemplary amine salts of Scheme 4 can be employed as starting materials in the synthetic methods herein. Appendix A also provides additional methods for synthesis of compounds of Formula I herein where the R group is an acylamino group. Scheme 6 herein provides exemplary carboxylic acid precursors for incorporation of various useful acyl amino groups into compounds of the disclosure wherein R is an acylamino group. In some embodiments, any of the exemplary carboxylic acids of Scheme 6 can be employed as starting materials in the synthetic methods herein. Synthetic methods herein are specifically illustrated for the preparation of cephalosporins, monobactams and carbapenems. These exemplary methods can be readily adapted to the preparation of cephamycins, carbacephems, oxacephems, penems, and oxapenems.
[0245] It will be appreciated that compounds of the disclosure can be prepared using methods exemplified herein or by routine adaptation of such methods with appropriate choice of starting materials. Methods exemplified herein can be used to prepare various cephem compounds, including cephalosporins, cephamycins and the like. Compounds with various Ri groups can be prepared from known precursors, such as the carboxylic acid precursors illustrated in the Scheme provided by know methods. Compounds with various -CH2-X groups can be prepared from known precursors by known methods. Methods provided herein can be routinely adapted to the synthesis of penems and oxapenems in view of synthetic methods for these compounds that are well-known in the art.
Figure imgf000108_0001
Figure imgf000109_0001
Y and Z are independently CH or N.
Synthesis Scheme 2: Synthesis of Monobactams of Formula I
Figure imgf000109_0002
Figure imgf000110_0001
Figure imgf000111_0001
Synthesis Scheme 3: Synthesis of Monobactams of Formula I
Figure imgf000111_0002
Figure imgf000112_0001
Figure imgf000113_0001
Figure imgf000114_0001
Figure imgf000115_0001
Synthesis Scheme 5: Synthesis of Carbapenems of Formula I
Figure imgf000115_0002
Figure imgf000116_0001
[0246] The term "pharmaceutically effective amount" refers to an amount effective in treating a bacterial infection, or a symptom or complication thereof, in a patient (human or other mammal) either by administration of a single compound of Formulas I- III, or a salt or solvate thereof or in combination with other agents. The pharmaceutically effective amount of a given compound when administered as the only active ingredient may differ from its pharmaceutically effective amount when administered with other active ingredients. It will be appreciated that the pharmaceutically effective amount of a compound may differ from that of a salt of the same compound.
[0247] The term “treating” includes the alleviation of symptoms of a particular disorder in a patient or a measurable improvement of a parameter associated with a particular disorder. Treating includes treatment to prevent a bacterial infection and to delay the progress of an infection. [0248] The term "prophylactically effective amount" refers to an amount of a compound or salt of the disclosure effective in preventing a bacterial infection in a patient. The compounds of the present disclosure are useful in the treatment of individuals infected by various bacteria (which include infections of combinations of different bacteria) for the prophylaxis of these individuals. It will be appreciated in the art, that individuals at risk for bacterial infection can be treated employing one or more of the compounds or salts of the disclosure to prevent or delay infection.
[0249] The term "patient" refers to any animal and more specifically to a mammal, including a human. In specific embodiments, the patient or subject is a human.
[0250] The number of carbons in a given group is designated herein using the terminology CX-CY, where X and Y are integers representing the lowest number and the highest number of carbons in the references group, as in C1 -C4 alkyl which refers to an alkyl group having 1 -4 carbon atoms. In a number of the formulas of this disclosure reference is made to the definition of variables in a given patent or patents or published U.S. patent application. In these instances, the variable definition from the patent document listed applies. In other cases, formula variables are specifically defined in the present specification and the definitions of such variables is defined herein or employs the broadest definition in the art of a given chemical moiety or group.
[0251] The terms “alkyl” or“alkyl group,” alone or in combination, refer to a monoradical of a straight chain or branched saturated hydrocarbon. Alkyl groups include straight-chain and branched alkyl groups. Unless otherwise indicated alkyl groups have 1 -12 carbon atoms (C1 -C12 alkyl groups) and preferred are those that contain 1 -6 carbon atoms (C1 -C6 alkyl groups) and more preferred are those that contain 1 -4 carbon atoms (C1 -C4 alkyl groups) and those that contain 1 -3 carbon atoms (C1 -C3 alkyl groups). Unless otherwise indicated alkyl groups are optionally substituted with one or more non-hydrogen substituents as described herein. However, any alkyl group designated herein can be unsubstituted. The designation of an alkyl group having a range of carbon atoms includes all isomers having that number of carbon atoms. Exemplary alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, s-butyl, t- butyl, n-pentyl, branched pentyl, n-hexyl, branched hexyl, all of which are optionally substituted. Substituted alkyl groups include fully halogenated or semihalogenated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms. Substituted alkyl groups include fully fluorinated or semifluorinated alkyl.
[0252] The term“alkoxy” refers to an -O-alkyl group, where alkyl is as defined above. Alkoxy groups include among others methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert butoxy. Alkoxy groups are optionally substituted.
[0253] The term“cycloalkyl,” alone or in combination, means an alkyl radical which contains at least one carbon ring. These groups may be monocyclic, bicyclic or tricyclic. Unless otherwise indicated a cycloalkyl contains from 3 to 12 carbons and the carbon ring contains 3-10 carbons and more preferably 3-7 carbon atoms. Examples of such cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups are optionally substituted.
[0254] The term“alkenyl,” alone or in combination, refers to a straight-chain or branched-chain mono-,di- or poly-unsaturated aliphatic hydrocarbon radical containing one or more double bonds and the specified number of carbon atoms, or where no number is specified, preferably from 2-10 carbon atoms, more preferably, from 2-6 carbon atoms and also from 2-4 carbon atoms. Unless specifically stated, all isomers of the given number of carbon atoms are included. Examples of alkenyl radicals include, but are not limited to, ethenyl, E- and Z-propenyl, isopropenyl, E- and Z-butenyl, E- and Z-isobutenyl, E- and Z-pentenyl, E- and Z-hexenyl, E,E-, E,Z-, Z,E- and Z,Z hexadienyl. Alkenyl groups are optionally substituted.
[0255] The term“alkenoxy” refers to an -O-alkenyl group, where alkenyl is defined above. Alkenoxy groups are optionally substituted.
[0256] The term“cycloalkenyl” means an alkyl radical which contains at least one carbon ring and at least one double bond. These groups may be monocyclic, bicyclic or tricyclic. Unless otherwise indicated a cycloalkyl contains from 3 to 12 carbons and the carbon ring contains 3-10 carbons and more preferably 3-7 carbon atoms. Examples of such cycloalkenyl groups include cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. Cycloalkenyl groups are optionally substituted.
[0257] The term“alkynyl,” alone or in combination, refers to a straight-chain or branched-chain aliphatic hydrocarbon radical containing one or more triple bonds and the specified number of carbon atoms, or where no number is specified, preferably from 2-10 carbon atoms, more preferably, from 2-6 carbon atoms and also from 2-4 carbon atoms. Specific alkynyl groups contain one triple bond or two triple bonds. Unless specifically stated, all isomers of the given number of carbon atoms are included. Examples of alkynyl radicals include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc. Alkynyl groups are optionally substituted. The term alkynoxy refers to an -O-alkynyl group, where alkynyl is defined above. Alkynoxy groups are optionally substituted.
[0258] An acyl group is an R'-CO- group where R' in general is a hydrogen, an alkyl, alkenyl or alkynyl, aryl, heterocyclyl, or heteroaryl group as described herein. In specific embodiments, R' is a substituted methylene group. In specific embodiments, acyl groups have 1 -20, 1 - 12, or 1 -6 carbon atoms and optionally 1 -3 heteroatoms, optionally one double bond or one triple bond. In specific embodiments, R is a C1 -C6 alkyl, alkenyl group cyclic configuration or a combination thereof, attached to the parent structure through a carbonyl functionality. Examples include acetyl, benzoyl, propionyl, isobutyryl, or oxalyl. The R' group of an acyl group is optionally substituted as described herein. When R' is hydrogen, the group is a formyl group. An acetyl group is a Chh- CO- group. Another exemplary acyl group is a benzyloxy group.
[0259] An acylalkoxy group is an alkoxy group as defined above substituted with an acyl group as defined above, e.g., -OCH2-COR. An alkyl group substituted with an acylalkoxy group is an acylalkoxyalkyl group.
[0260] An acyloxy group is an acyl group as defined above bonded to an oxygen, e.g., -O-COR. An alkyl group substituted with an acyloxy group is an acyloxyalkyl group.
[0261] An acylamino group is an R'-CO-NH- group, where R' is as defined for the acyl group above. A number of acylamino groups are known in the art as suitable for use in cephem and penem antibiotics.
[0262] An amino group is -NH2. An alkylamino group is -NHR', where R' is an alkyl group, preferably which is a C1 -C4 alkyl or a C1 -C3 alkyl. An alkylamino group is -NR2', where R' is an alkyl group, preferably which is a C1 -C4 alkyl or a C1 -C3 alkyl. Amino, alkylamino and dialkyl amino groups can be protonated or quaternized, for example, with bonding of another alkyl group resulting in a positively charged amino, alkyl amino, dialkyamino, trialky amino or a quaternary amino group. Certain compounds herein have cyclic amino groups where the nitrogen is in a ring. The ring may otherwise contain only carbon ring atoms or the ring may contain an additional heteroatom, such as 0, N or S. The ring may be saturated (containing no double bonds), partially unsaturated or fully unsaturated. The nitrogen in the ring can have the form - NH-, or =N- or be a ring member of an aromatic ring (e.g., such as a pyridine). As is illustrated in formulas herein, the ring N can be protonated or alkylated (with a C1 -C4 alkyl group) to form a positively charged ring amino group.
[0263] The term“aryl,” alone or in combination, refers to a carbocyclic aromatic radical (such as phenyl or naphthyl) containing the specified number of carbon atoms. If not specified aryl groups contain from 6-15 carbon atoms, preferably from 6-10 carbon atoms, and particularly contain from 6-10 ring carbons. Aryl groups unless otherwise stated are optionally substituted among others with one or more substituents selected from alkyl, alkoxy, nitro, halogen, (for example, chloro), amino, alkyl amino, dialkylamino, carboxylate and hydroxy. In specific embodiments, aryl groups are optionally substituted phenyl groups. Aryl groups may contain two rings that are fused (naphthyl) or two rings which are bonded together by a C-C bond (biphenyl). Examples of aryl groups include, among others, phenyl, p-tolyl, 4-hydroxyphenyl, 1 -naphthyl, 2-naphthyl, indenyl, indanyl, azulenyl, fluorenyl, and anthracenyl.
[0264] The term“arylalkyl,” alone or in combination, refers to an alkyl group as described above which is substituted with an aryl group as defined above. In specific embodiments, the aryl group is an optionally substituted phenyl, naphthyl or biphenyl group. In specific embodiments, the arylalkyl is a phenalkyl group. Specific phenalkyl groups are benzyl and phenethyl groups.
[0265] The term“heterocyclyl” or“heterocyclic” refers to monoradical having a ring of a specified number of ring atoms, where the ring atoms include one or more heteroatoms (N, O, S) or heteroatom groups (e.g., -NH-, or -N(alkyl)-). More specifically, the term includes groups having a stable 3-7 membered monocyclic heterocyclic ring or an 8-1 1 membered bicyclic heterocyclic ring. The ring can be saturated, or partially or fully unsaturated, and may be optionally benzofused, if monocyclic and is optionally substituted unless otherwise stated on one or more carbon atoms by halogen, alkyl, alkoxy, oxo (=0) or on a secondary nitrogen atom by alkyl, phenyl or phenylalkyl. A number of heterocyclic groups are exemplified in the specification. Each heterocycle consists of one or more carbon atoms and from one to four heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur and oxidized forms thereof. Heterocycles include 5-7 membered monocyclic heterocycles and 8-10 membered bicyclic heterocycles.
[0266] The term“heteroaryl” refers to a group having at least one aromatic ring wherein the ring contains at least one heteroatom or heteroatom group, as defined above. More specifically, the term refers to stable 5-6 membered monocyclic or 8-1 1 membered bicyclic aromatic heterocycles where heterocycles is as defined above. Non-limiting examples of such groups include imidazolyl, quinolyl, isoquinolyl, indolyl, indazolyl, pyridazyl, pyridyl, pyrrolyl, pyrazolyl, pyrazinyl, quinoxolyl, pyranyl, pyrimidinyl, furyl, thienyl, triazolyl, thiazolyl, carbolinyl, tetrazolyl, benzofuranyl, thiamorpholinyl sulfone, oxazolyl, benzoxazolyl, benzimidazolyl, benzthiazolyl, oxopiperidinyl, oxoppyrrolidinyl, oxoazepinyl, azepinyl, isoxazolyl, isothiazolyl, furazanyl, thiazolyl, thiadiazolyl, and oxathiolyl.
[0267] A carbocyclyl group is a group having one or more saturated or unsaturated carbon rings. Carbocyclyl groups, for example, contain one or two double bonds. One or more carbons in a carbocyclic ring can be -CO- groups. Carbocyclyl groups include those having 3-12 carbon atoms, and optionally replacing 1 or 2 carbon atoms with a - CO- group and optionally having 1 , 2 or 3 double bonds. Carbocyclyl groups include those having 5-6 ring carbons. Carbocyclyl groups can contain one or more rings each of which is saturated or unsaturated. Carbocyclyl groups include bicyclic and tricyclic groups. Preferred carbocyclic groups have a single 5- or 6- member ring. Carbocyclyl groups are optionally substituted as described herein. Specifically, carbocyclic groups can be substituted with one or more alkyl groups. Carbocyclyl groups include among others cycloalkyl and cycloalkenyl groups.
[0268] A styryl group or moiety is a group or moiety in which a vinyl (ethylene) group is bonded to a phenyl ring. The group or moiety may be substituted with one or more non-hydrogen substituents on the phenyl ring or most generally on the vinyl moiety. In formulas of monobactams herein, where M is M3 and n is 1 and L is ethylene (vinyl), the styryl group can be cis or trans or with respect to the monobactam ring or a mixture of cis and trans isomers. The cis and trans configurations can also be designated the E or Z configuration (or a mixture of E/Z configurations), where one of ordinary skill in the art understand how to name E and Z configurations for a given structure. The E and Z configurations are exemplified in styryl groups having formulas:
Figure imgf000122_0001
configuration, where R5-R9 are optional substituents as defined for the B ring herein and one of R5-R9 is -CH2-X. The vinyl group can be written as a cross double bond to indicate that the double bond is a mixture of the E and Z configurations (cis and trans isomers). Compounds herein can be a mixture of isomers having the E or Z configuration at the styryl group. In some embodiments, monobactams herein include variants of the styryl group in which one-four of the carbons of the phenyl ring are replaced with nitrogens, for example, a -vinylpyridyl group. Such heteroaryl styryl variants can also be in the cis or trans configuration (or E and Z configurations) with respect to the monobactam ring. These vinylheteroaryl groups can be in the cis or trans configuration or can be a mixture of cis/trans configurations, e.g., the compound would be a mixture of cis/trans (or E/Z isomers).
[0269] The term“cephem” refers generally to the ring system:
Figure imgf000122_0002
where ring atoms are numbered as indicated and T is -S-, -SO-, -SO2-, -CHR17-, or -0-. This numbering is used throughout for labeling of ring substituents. It will be appreciated by one of ordinary skill in the art that different ring numbering conventions may be employed in the art. [0270] The term“penem” refers generally to the ring system:
Figure imgf000123_0001
where ring atoms are numbered as indicated and U is -S-, -CHR17- or -0-. This numbering is used throughout for labeling of ring substituents. It will be appreciated by one of ordinary skill in the art that different ring numbering conventions may be employed in the art. It is noted that the term penem is used generically herein. The term penem is also used in the art to refer to compounds of this ring system where U is -S-.
[0271] Carboxylate esters (-CO2-E) include those where E is an optionally substituted C1 -C6 or C1 -C3 alkyl (optional substation includes among others substitution with one or more halogen), an optionally substituted phenyl or phenyalkyl group (where substitution includes among others ring substitution with one or more halogen or with a nitro group). Carboxylate esters further include those in which E is an alkoxyalkyl group, more specifically where the alkoxy and the alkyl group is C1 -C6 or C1 -C3 alkoxy and C1 -C6 or C1 -C3 alkyl, e.g. , methyoxym ethyl, ethoxyethyl, methoxyethyl and the like). Carboxylate esters (-CO2-E) include those where E is an acylalkoxyalkyl group, more specifically where the acyl, alkoxy and alkyl groups are C1 - C6 or C1 -C3 acyl, alkoxy or alkyl groups.
[0272] R1-CO-NH- in formulas herein is an acylamino group. In general, R1-CO-NH- can be any pharmaceutically acceptable A such group that is known in the art to be compatible with a cephem antibiotic. A number of examples of Ri useful groups are provided in the specification. Additional useful Ri groups are known in the art. In specific embodiments, useful Ri groups are substituted methylene groups. In other embodiments, useful Ri groups are substituted oximes. In other specific embodiments, useful Ri groups are substituted vinyl ether groups.
[0273] All references throughout this application, for example, patent documents including issued or granted patents or equivalents; patent application publications; and non-patent literature documents or other source material; are hereby incorporated by reference herein in their entireties, as though individually incorporated by reference.
[0274] When a group of substituents is disclosed herein, it is understood that all individual members of that group and all subgroups, including any isomers and enantiomers of the group members, and classes of compounds that can be formed using the substituents described are disclosed separately.
[0275] Every formulation or combination of components described or exemplified can be used to practice the disclosure, unless otherwise stated. Specific names of compounds are intended to be exemplary, as it is known that one of ordinary skill in the art can name the same compounds differently. When a compound is described herein such that a particular isomer or enantiomer of the compound is not specified, for example, in a formula or in a chemical name, that description is intended to include each isomers and enantiomer of the compound described individual or in any combination.
[0276] One of ordinary skill in the art will appreciate that methods, device elements, starting materials, and synthetic methods other than those specifically exemplified can be employed in the practice of the disclosure without resort to undue experimentation. All art-known functional equivalents, of any such methods, device elements, starting materials, and synthetic methods are intended to be included in this disclosure. Whenever a range is given in the specification, for example, a temperature range, a time range, or a composition range, all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the disclosure.
[0277] The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the disclosure claimed. Thus, it should be understood that although the present disclosure has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this disclosure. [0278] Abbreviations: acetate (Ac); ethyl acetate (AcOET); acetonitrile (ACN); broad (br); butyl (Bu); d (doublet); dibenzylideneacetone (dba); dicholormethane (DCM); diisopropyl azodicarboxylate (DIAD); N,N-diisopropylethylamine (DIPEA); 4- dimethylaminopyridine (DMAP); 1 ,2-dimethylethylenediamine (DMEDA); dimethylformamide (DMF); di-fe/f-butyl azodicarboxylate (DTBAD); ethyl (Er); proton (in NMR) (H); Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium (HATU); hertz (Hz); imidazole (Imid.); coupling constant (J); multiplet (m); meta- chloroperoxybenzoic acid (mCPBA); methyl (Me); methanesulfonyl (Ms); N- methylmorpholine (MNN); N-methyl-pyrrolidone (MNP); Nuclear Magnetic Resonance (NMR); parts per million (ppm); quartet (q); quintet (qnt); room temperature (rt); singlet (s); saturated (sat.); triplet (t); tetra-n-butylammonium fluoride (TBAF); tert- butyldimethylsilyl (TBS or TBDMS); triethylsilyl (TES); trifluoromethanesulfonyl (Tf); trifluoroacetic acid (TFA); and tetrahydrofuran (THF).
Examples
Example 1 : Preparation of Exemplary Phenyl Cephalosporins of Formula I:
Compounds 482 487 501 502 503
Figure imgf000125_0001
A. Preparation of [4-(tributylstannyl)phenyl1methanol (2)
[0279] Compound 2 is prepared by a literature procedure [25] More specifically, 4-bromobenzyl alcohol 1 (1 .400 g, 7.485 mmol, 1.000 equiv) is dissolved in anhydrous THF (21 ml_, 0.35 M) at room temperature(rt), under anhydrous conditions. At - 78 °C, a 2.5 M solution of n-BuLi in hexanes (6.14 ml_, 15.3 mmol, 2.05 equiv) is added dropwise to the THF solution with stirring. The resulting mixture solidified after addition of n-BuLi. A solution of tributyltin chloride (4.995 g, 15.35 mmol, 2.05 equiv) in anhydrous THF (5 ml_, 3 M) is then slowly added to the reaction, which is thereafter is warmed to rt. The white solid that had formed dissolved upon warming. The resulting yellow solution is stirred at rt for 1 hour and 40 minutes. Reaction is then quenched by slow addition of saturated ammonium chloride solution (10 ml_). The resulting quenched reaction is stirred at rt for 1 hour and 10 minutes. The mixture is then transferred into a separatory funnel and brine (ca. 40 ml_) and AcOEt (ca. 30 ml_) are added. The aqueous layer is separated from the organic layer and is extracted with AcOEt (ca. 30 ml_). The combined organic layers are dried over magnesium sulfate, filtered and evaporated under reduced pressure. The colorless oil obtained is chromatographed on silica gel (Biotage SNAP, 50 g) eluting with 0-20% AcOEt/Hexanes. Product 2 is obtained as colorless oil (2.522 g, 6.350 mmol, 85% yield).
[0280] 1H NMR (300 MHz, CDC ) d (ppm) 7.60-7.40 (m, 2H), 7.40-7.30 (m, 2H), 4.69 (d, 2H, J = 5.7 Hz), 1.65-1.5 (m, 6H), 1.45-1.25 (m, 6H), 1.15-1.00 (m, 6H), 0.95- 0.80 (m, 9H).
Figure imgf000126_0001
3 4
B. Preparation of (6R,7R)-benzhvdryl 8-oxo-7-(2-phenylacetamido)-3-
Figure imgf000126_0002
(((trifluoromethyl)sulfonyl)oxy)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate (4)
[0281] Preparation of compound 4 is adapted from U.S. Patent 4,870,168. Under anhydrous conditions, (6R,7R)-benzhydryl 3-hydroxy-8-oxo-7-(2-phenylacetamido)-5- thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate 3 (2.300 g, 4.595 mmol, 1.000 equiv) is dissolved in anhydrous DCM (77 ml_, 0.06 M). To the pale-yellow solution, at - 20 °C, diisopropylethylamine (0.800 ml_, 4.595 mmol, 1.000 equiv) is added dropwise. At - 20 °C, trifluoromethanesulfonyl anhydride (0.926 ml_, 5.514 mmol, 1.200 equiv) is added dropwise. The resulting white suspension is stirred at - 20 °C for 1 hour. The suspension is diluted with DCM (300 ml_) and the resulting organic layer is mixed with 0.25 N HCI (80 ml_) and then with water (150 ml_). The washed organic layer is dried over magnesium sulfate, filtered and solvent is removed under reduced pressure. The product 4 is obtained as pale-orange solid (2.801 g, 4.428 mmol, 96% yield). This product is used without further purification in the next step.
[0282] 1H NMR (300 MHz, CDC ) d (ppm) 7.45-7.20 (m, 15H), 6.99 (s, 1 H), 5.98 (d, 1 H, J = 9.0 Hz), 5.95-5.85 (m, 1 H), 5.05 (d, 1 H, J = 4.8 Hz), 3.77 (d, 1 H, J = 18.0 Hz), 3.70 (d, 1 H, J = 16.5 Hz), 3.62 (d, 1 H, J = 16.5 Hz), 3.43 (d, 1 H, J = 18.6 Hz).
Figure imgf000127_0001
C. Preparation of (6R,7R)-benzhvdryl 3-(4-(hvdroxymethyl)phenyl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate (5)
[0283] Preparation of compound 5 is adapted from U.S. Patent 4,870, 168. Under anhydrous conditions, the triflate 4 (2.800 g, 4.426 mmol, 1 .000 equiv) is dissolved in anhydrous NMP (32 mL, 0.12 M). To this orange solution, the stannane 2 (2.1 10 g, 5.31 1 mmol, 1 .200 equiv) is added at rt as a solution in anhydrous NMP (5 mL, 1 M). At rt, zinc chloride (1 .206 g, 8.852 mmol, 2.000 equiv) and tri(2-furyl)phosphine (0.206 g, 0.885 mmol, 0.200 equiv) were added to the mixture. Nitrogen gas is then bubbled into the mixture for five minutes. Thereafter, at rt, palladium bis(dibenzylidene acetone) (0.255 g, 0.443 mmol, 0.100 equiv) is added to the reaction mixture in one portion. The resulting dark brown solution is heated at 55 °C for 20 hours. The mixture is then diluted in ethyl acetate (150 mL) and washed with saturated ammonium chloride solution (2 x 150 mL). The collected aqueous washes are extracted with ethyl acetate (100 mL). The combined organic layers are evaporated under reduced pressure. The residue is dissolved in acetonitrile (150 mL) and this solution is washed with hexanes (100 mL). Solvent is evaporated under reduced pressure. The dark orange-brown oil obtained is purified by column chromatography on silica gel (Biotage SNAP 100 g) eluting with 20- 65% AcOEt/Hexanes. The product is further purified by a second column chromatography on silica gel (Biotage SNAP 50 g) eluting with 20-60% AcOEt/Hexanes. Product 5 is obtained as orange solid (1 .275 g, 2.159 mmol, 49% yield). [0284] 1H NMR (500 MHz, CDCb) d (ppm) 7.45-7.38 (m, 2H), 7.37-7.32 (m, 1 H), 7.32-7.23 (m, 5H), 7.23-7.17 (m, 5H), 7.16-7.1 1 (m, 2H), 7.05 (d, 2H), 6.93 (d, 2H), 6.80 (s, 1 H), 6.04 ( br d, 1 H, J = 9.5 Hz), 5.91 (dd, 1 H, J = 8.5 Hz, J = 5.0 Hz), 5.07 (d, 1 H, J = 5.0 Hz), 4.59 (d, 2H, J = 5.5 Hz), 3.72 (d, 1 H, J = 16.0 Hz), 3.64 (d, 1 H, J = 16.0 Hz), 3.60 (s, 2H).
Figure imgf000128_0001
D. Preparation of (6R,7R)-benzhvdryl 3-(4-(chloromethyl)phenyl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate (6)
[0285] Under anhydrous conditions, alcohol 5 (1 .270 g, 2.150 mmol, 1 .000 equiv) is dissolved in anhydrous DCM (18 ml_, 0.12 M). At 0 °C, methanesulfonyl chloride (0.250 ml_, 3.23 mmol, 1 .50 equiv) is added dropwise, followed by dropwise addition of /V-methylmorpholine (NMM) (0.307 ml_, 2.80 mmol, 1 .30 equiv). The resulting orange solution is stirred at 0 °C for 5 minutes and then at rt for 1 hour and 20 minutes. Solvent is evaporated to dryness and the resulting pale-brown solid is dissolved in anhydrous DMF (1 1 ml_, 0.20 M) and lithium chloride (0.182 g, 4.30 mmol, 2.00 equiv) is added in one portion. The brown solution is then stirred at rt for 2 hours and 45 minutes. The mixture is diluted in ethyl acetate (60 ml_) and water (50 ml_) is added, followed by a few ml_ of saturated ammonium chloride solution. The organic layer is then washed with water (40 ml_) and brine, dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The resulting brown solid is chromatographed on silica gel (Biotage SNAP 25 g) eluting with 10-40% AcOEt/Hexanes gradient. Product 6 is obtained as off-white solid (1.017 g, 1 .670 mmol, 78% yield).
[0286] 1H NMR (500 MHz, CDC ) d (ppm) 7.42-7.37 (m, 2H), 7.37-7.33 (m, 1 H), 7.32-7.25 (m, 5H), 7.24-7.13 (m, 7H), 7.05-7.00 (m, 2H), 6.95-6.90 (m, 2H), 6.81 (s, 1 H), 6.05 (br d, 1 H, J = 9.0 Hz), 5.92 (dd, 1 H, J = 9.0 Hz, J = 5.0 Hz), 5.06 (d, 1 H, J = 5.0 Hz), 4.50 (d, 1 H, J = 1 1 .5 Hz), 4.46 (d, 1 H, J = 1 1 .5 Hz), 3.72 (d, 1 H, J = 16.0 Hz), 3.64 (d, 1 H, J = 16.0 Hz), 3.60 (d, 1 H, J = 19.0 Hz), 3.56 (d, 1 H, J = 19.0 Hz).
Figure imgf000129_0001
8 9
E. Preparation of (6R,7R)-benzhvdryl 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-3- (4-(chloromethyl)phenyl)-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate (10)
[0287] Under anhydrous conditions, phosphorus pentachloride (2.076 g, 9.968 mmol, 6.000 equiv) is suspended in anhydrous DCM (1 1 ml_, 0.90 M). At - 5 °C, pyridine (0.806 ml_, 9.97 mmol, 6.00 equiv) is added dropwise. To the resulting white suspension, the chloride 6 (1 .012 g, 1 .661 mmol, 1 .000 equiv) is added dropwise as a solution in anhydrous DCM (12 ml_, 0.14 M), at - 5 °C. The resulting pale-yellow suspension is stirred between - 5 °C and 0 °C for 1 .5 hours. At 0 °C, anhydrous methanol (6.73 ml_, 166 mmol, 100 equiv) is added dropwise to the suspension and the resulting solution is stirred at 0 °C for 15 minutes and then at rt for 10 minutes. The solution is then diluted in DCM (80 ml_) and saturated sodium bicarbonate solution (80 ml_) is added carefully. The separated organic layer is washed with dilute (ca. 0.1 M) sodium bicarbonate solution, dried over magnesium sulfate, filtered and solvent is evaporated until 10-15 mL of the solution of free amine 7 remained.
[0288] In parallel, a solution of acid chloride 9 is prepared as follows:
[0289] Under anhydrous conditions, phosphorus pentachloride (0.478 g, 2.29 mmol, 1 .38 equiv) is suspended in anhydrous DCM (9.6 mL, 1 .4 M) at rt. The white suspension is stirred at 0 °C for 10 minutes. At 0 °C, the carboxylic acid 8 (0.821 g, 1 .91 mmol, 1 .15 equiv) is added in one portion. The heterogeneous mixture containing acid chloride 9 is stirred, between - 5 ° and 0 °C for 45 minutes.
[0290] To the latter acid chloride solution is added the free amine solution (ca. 10- 15 mL) dropwise at -5 °C. The resulting pale-yellow solution is stirred between -5 °C and rt for 1 hour and 50 minutes. The mixture is then diluted in DCM (45 mL), washed first with dilute ammonium chloride solution (50 mL), then with 0.5 N HCI solution (2 x 50 mL) and thereafter with dilute (0.1 M) sodium bicarbonate solution (40 mL). The washed milky organic layer is dried over sodium sulfate, filtered and evaporated under reduced pressure. The off-white solid obtained is purified by reverse phase chromatography (Biotage C18 60 g) eluting with 5-100% ACN/H2O. The product 10 is obtained as off-white solid (0.797 g, 0.883 mmol, 53% yield).
[0291] 1H NMR (500 MHz, CDCL) d (ppm) 8.12 (d, 1 H, J = 8.0 Hz), 7.35 (s, 1 H), 7.32-7.24 (m, 3H), 7.24-7.14 (m, 7H), 7.07-7.02 (m, 2H), 6.98-6.93 (m, 2H), 6.83 (s, 1 H), 6.09 (dd, 1 H, J = 8.5 Hz, J = 4.5 Hz), 5.18 (d, 1 H, J = 5.0 Hz), 4.51 (d, 1 H, J = 12.0 Hz), 4.48 (d, 1 H, J = 12.5 Hz), 3.64 (s, 2H), 1 .66 (s, 3H), 1 .62 (s, 3H), 1 .54 (s, 9H), 1 .43 (s, 9H).
Figure imgf000131_0001
F. Preparation of (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-(4-(chloromethyl)phenyl)-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-ene-2-carboxylic acid 1 1 (504)
[0292] Under nitrogen atmosphere, chloride 10 (0.120 g, 0.133 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (2.7 ml_, 0.050 M). At 0 °C, anisole (0.145 ml_, 1 .33 mmol, 10.0 equiv) is added slowly, followed by slow addition of TFA (2.7 ml_, 0.050 M). The colorless solution is stirred at 0 °C for 5 minutes becoming a pale-yellow solution, which is then stirred at rt for 3 hours and 45 minutes. The solution is evaporated to dryness. The orange solid obtained is triturated three times in 50/50 v/v diethyl ether/hexanes (3 x 4 ml_). Remaining traces of solvent are evaporated. The product 11 is obtained as pale-orange solid (0.084 g, quantitative yield), which is used without further purification in the next step.
[0293] 1H NMR (500 MHz, Acetone-de) d (ppm) 7.47-7.42 (m, 2H), 7.38-7.32 (m, 2H), 6.96 (s, 1 H), 6.12-6.04 (m, 1 H), 5.34 (d, 1 H, J = 5.0 Hz), 4.72 (s, 2H), 3.90 (d, 1 H, J = 19.0 Hz), 3.83 (d, 1 H, J = 19.0 Hz), 1 .66 (s, 3H), 1 .62 (s, 3H). m/z = 580 [M+H]+.
Figure imgf000131_0002
11 13 G. Preparation of 2-(4-((6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-en-3-yl)benzyl)-6,7-dihvdroxyisoquinolin-2-ium 13 (482)
[0294] Under nitrogen atmosphere, 6,7-dihydroxyisoquinolin-2-ium bromide 12
(0.0075 g, 0.031 mmol, 1 .2 equiv) and potassium carbonate (0.0075 g, 0.054 mmol, 2.1 equiv) and compound 11 (0.015 g, 0.026 mmol, 1 .0 equiv) are suspended in anhydrous DMF (0.26 ml_, 0.10 M). To the brown suspension, containing white solids, potassium iodide (0.0043 g, 0.026 mmol, 1 .0 equiv) is added. The reaction mixture is stirred at rt, under nitrogen atmosphere for 6 hours. TFA (3 drops) are added and the resulting yellow solution is evaporated under reduced pressure. The resulting yellow suspension is chromatographed on a reverse phase column (Teledyne GOLD C18 5.5 g), eluting with a 5-80% ACN/H2O gradient (+ 0.1 % TFA). Fractions containing the desired product 13 were lyophilized and the product 13 is obtained as a fluffy white solid (0.003 g, 0.004 mmol, 16% yield).
[0295] 1H NMR (500 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 9.59 (s, 1 H), 8.43 (dd, 1 H, J = 6.5 Hz, J = 1 .5 Hz), 8.14 (d, 1 H, J = 7.0 Hz), 7.70 (s, 1 H), 7.56-7.50 (m, 2H), 7.40-7.34 (m, 2H), 7.07 (s, 1 H), 5.99 (d, 1 H, J = 5.0 Hz), 5.92 (s, 2H), 5.30 (d, 1 H, J = 5.0 Hz), 3.82 (d, 1 H, J = 19.0 Hz), 3.76 (d, 1 H, J = 19.0 Hz), 1 .61 (s, 3H), 1 .58 (s, 3H). m/z = 705.4 [M]+, 353.3 [M+H]2+.
Figure imgf000132_0001
H. Preparation of 1 -(4-((6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-en-3-yl)benzyl)-5,6-dihvdroxy-2-methyl-2H-indazol-1 -ium 15 (487)
[0296] Under nitrogen atmosphere, 2-methyl-2/-/-indazol-1 -ium bromide 14 (0.331 g, 1 .37 mmol, 8.00 equiv) and potassium carbonate (0.130 g, 0.939 mmol, 5.50 equiv) are suspended in anhydrous DMF (1 .71 ml_, 0.10 M). The dark-yellow suspension, containing white solids, is stirred at rt for 10 minutes. Then, compound 11 (0.099 g, 0.171 mmol, 1 .00 equiv) is added, quickly followed by potassium iodide (0.028 g, 0.171 mmol, 1 .00 equiv). The resulting cloudy dark green mixture is immersed in a preheated oil bath (50 °C) and is stirred at 50 °C for 1 .5 hours. The heating is stopped and 10 drops of TFA are added, followed by ca. 2 ml_ of water. This solution is charged onto a reverse phase column (Biotage C18 60 g) and eluted with a 5-100% ACN/H2O gradient. Fractions containing the desired product were lyophilized and the product 15 is obtained as a fluffy white solid (0.054 g, 0.076 mmol, 45% yield).
[0297] 1H NMR (500 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 8.79 (s, 1 H), 7.34-7.29 (m, 2H), 7.28 (s, 1 H), 7.22 (s, 1 H), 7.20-7.14 (m, 2H), 7.07 (s, 1 H), 6.02-5.95 (s + d, 2 + 1 H), 5.29 (d, 1 H, J = 5.0 Hz), 4.30 (s, 3H), 3.81 (d, 1 H, J = 18.5 Hz), 3.76 (d, 1 H, J = 18.5 Hz), 1 .64 (s, 3H), 1 .61 (s, 3H). m/z = 708.7 [M]+, 354.9 [M+H]2+.
Figure imgf000133_0001
I. Preparation of (6R,7R)-benzhvdryl 7-((Z)-2-(2-((tert- butoxycarbonyl)amino)thiazol-4-yl)-2-(methoxyimino)acetamido)-3-(4- (chloromethyl)phenyl)-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 18
[0298] Under anhydrous conditions, phosphorus pentachloride (0.492 g, 2.36 mmol, 8.00 equiv) is suspended in anhydrous DCM (2.0 ml_, 0.15 M). At 0 °C, and pyridine (0.191 ml_, 2.36 mmol, 8.00 equiv) is added dropwise. To the resulting white suspension, the chloride 6 (0.180 g, 0.296 mmol, 1 .00 equiv) is added dropwise as a solution in anhydrous DCM (2.0 ml_, 0.14 M), at - 10 °C. The pale-yellow suspension is stirred between - 10 °C and - 5 °C for 1 hour and 10 minutes. The yellow suspension is then stirred at 0 °C for 0.5 hour. At - 70 °C, anhydrous methanol (0.95 ml_, 30 mmol, 100 equiv) is added dropwise and the solution is stirred at - 70 °C for 15 minutes and then at 0 °C for 15 minutes. The mixture is evaporated to dryness and dried under high vacuum. The resulting yellow oil is taken up in DCM (25 ml_) and this solution is carefully washed with saturated sodium bicarbonate solution (25 ml_). The aqueous wash is extracted using DCM (15 ml_) and the combined organic layers are washed with brine (30 ml_). The yellow organic solution containing free amine 7 is dried over magnesium sulfate, filtered and evaporated under reduced pressure until around 5 mL of solution remained.
[0299] In parallel, a solution of the methanesulfonic anhydride 17 is prepared as follows:
[0300] Under anhydrous conditions, carboxylic acid 16 (0.102 g, 0.340 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (2.3 ml_, 0.15 M) at rt. At 0 °C, methanesulfonyl chloride (0.034 ml_, 0.44 mmol, 1 .3 equiv) is added dropwise, followed by N- methylmorpholine (0.037 ml_, 0.34 mmol, 1.0 equiv). The homogeneous mixture containing compound 17 is stirred at 0 °C for 5 minutes and then at rt for 25 minutes.
[0301] To the latter methanesulfonyl anhydride solution is added the free amine solution (ca. 5 mL) dropwise at 0 °C. The resulting pale-yellow solution is stirred at 0 °C for 15 minutes and rt for 50 minutes. The mixture is diluted in DCM (25 ml_), washed with saturated ammonium chloride solution (20 ml_), then with 0.5 N HCI solution (20 ml_) and dilute (0.1 M) sodium bicarbonate solution (20 ml_). The yellow organic solution is dried over magnesium sulfate, filtered and evaporated under reduced pressure. The resulting yellow oil is purified by normal phase chromatography (Biotage SNAP 25 g) eluting with 20-60% AcOEt/Hexanes. The product 18 is obtained as off-white solid (0.182 g, 0.235 mmol, 80% yield).
[0302] 1H NMR (500 MHz, CDCIs) d (ppm) 8.18 (br s, 1 H), 7.36 (s, 1 H), 7.34-7.26 (m, 3H), 7.24-7.16 (m, 7H), 7.14-7.05 (m, 3H), 6.98-6.94 (m, 2H), 6.84 (s, 1 H), 6.07 (dd, 1 H, J = 9.0 Hz, J = 5.0 Hz), 5.20 (d, 1 H, J = 5.0 Hz), 4.51 (d, 1 H, J = 1 1 .5 Hz), 4.48 (d, 1 H, J = 12.0 Hz), 4.12 (s, 2H), 3.67 (s, 3H), 1 .56 (s, 9H).
Figure imgf000135_0001
18 19
J. Preparation of (6R7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2- (methoxyimino)acetamido)-3-(4-(chloromethyl)phenyl)-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-ene-2-carboxylic acid 19 (501 )
[0303] Under nitrogen atmosphere, compound 18 (0.180 g, 0.232 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (3.9 ml_, 0.060 M). At 0 °C, anisole (0.177 ml_, 1 .63 mmol, 7.00 equiv) is added slowly, followed by slow addition of TFA (3.9 ml_, 0.060 M). The resulting yellow organic solution is stirred at 0 °C for 3 hours. The solution is evaporated to dryness. The resulting yellow solid is triturated in diethyl ether (4 x 3 ml_). Remaining traces of solvent are evaporated and hexanes (3 ml_) is added and evaporated. The solid is then dried under high vacuum. The product 19 is obtained as pale-green solid (0.134 g, quantitative yield), which is used without further purification in the next step.
[0304] 1H NMR (500 MHz, Acetone-de) d (ppm) 8.55 (br s, 1 H), 7.48-7.43 (m, 2H), 7.38-7.33 (m, 2H), 6.97 (br s, 1 H), 6.05-5.98 (m, 1 H), 5.32 (d, 1 H, J = 4.0 Hz), 4.72 (s, 2H), 3.95 (s, 3H), 3.88 (d, 1 H, J = 20 Hz), 3.82 (d, 1 H, J = 20 Hz).
Figure imgf000135_0002
19 20 K. Preparation of 2-(4-((6R 7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-
Figure imgf000136_0002
(methoxyimino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-en-3- yl)benzyl)-6,7-dihvdroxyisoquinolin-2-ium 20 (502)
[0305] Under nitrogen atmosphere, 6,7-dihydroxyisoquinolin-2-ium bromide 12 (0.019 g, 0.079 mmol, 2.0 equiv) and potassium carbonate (0.01 1 g, 0.079 mmol, 2.0 equiv) are suspended in anhydrous DMF (0.39 ml_, 0.10 M), at rt. Compound 19 (0.020 g, 0.039 mmol, 1 .0 equiv) is added, quickly followed by potassium iodide (0.013 g, 0.079 mmol, 2.0 equiv). The resulting brown suspension is stirred at rt, under nitrogen atmosphere for 4.5 hours. About 0.5 mL of water is then added to obtain a clear brown solution. 2 drops of TFA (2 drops) are added and the resulting solution is chromatographed on a reverse phase column (Teledyne GOLD C18 5.5 g), eluting with a 5-80% ACN/H2O gradient (+ 0.1 % TFA). Fractions containing the desired product were lyophilized and the product 20 is obtained as a fluffy white solid (0.010 g, 0.016 mmol, 40% yield).
[0306] 1H NMR (500 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 9.59 (s, 1 H), 8.46-8.41 (m, 2H), 8.14 (d, 1 H, J = 7.0 Hz)), 7.70 (s, 1 H), 7.56-7.50 (m, 2H), 7.39-7.33 (m, 2H), 7.02 ( br s, 1 H), 5.94 (d, 1 H, J = 5.0 Hz), 5.92 (s, 2H), 5.28 (d, 1 H, J = 5.0 Hz), 3.98 (s, 3H), 3.80 (d, 1 H, J = 19 Hz), 3.76 (d, 1 H, J = 19 Hz) m/z = 633.5 [M]+, 317.3 [M+H]2+.
Figure imgf000136_0001
L. Preparation of 1 -(4-((6R7F?)-7-((Z)-2-(2-aminothiazol-4-yl)-2- (methoxyimino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-en-3- yl)benzyl)-5,6-dihvdroxy-2-methyl-2/-/-indazol-1 -ium 21 (503)
[0307] Under nitrogen atmosphere, 2-methyl-2/-/-indazol-1 -ium bromide 14 (0.1 19 g, 0.492 mmol, 10.00 equiv) and potassium carbonate (0.041 g, 0.295 mmol, 6.00 equiv) are suspended in anhydrous DMF (0.49 mL, 0.10 M). The yellow suspension, containing white solids, is stirred at rt for 5 minutes. Then, the compound 19 (0.025 g, 0.049 mmol, 1 .0 equiv) is added, quickly followed by potassium iodide (0.025 g, 0.15 mmol, 3.0 equiv). The resulting cloudy yellow mixture is immersed in a preheated oil bath (40 °C) and is stirred at 40 °C for 4.5 hours. The heating is stopped and 0.5 mL of water is added followed by 3 drops of TFA. This solution is charged onto a reverse phase column (Biotage C18 12 g) and is eluted with a 5-100% ACN/H2O (+ 0.1 % TFA) gradient. Fractions containing the desired product are lyophilized and product 21 is obtained as a fluffy white solid (0.021 g, 0.033 mmol, 67% yield).
[0308] 1H NMR (500 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 8.81 (s. 1 H), 7.36-7.31 (m, 1 H), 7.29 (s, 1 H), 7.24 (s, 1 H), 7.22-7.17 (m, 2H), 7.00 (br s, 1 H), 6.01 (s, 2H), 5.95 (d, 1 H, J = 5.0 Hz), 5.28 (d, 1 H, J = 4.5 Hz), 4.32 (s, 3H), 3.97 (br s, 3H), 3.78 (s, 2H). m/z = 636.1 [M]+, 318.6 [M+H]2+.
Example 2: Preparation of Exemplary 2-Pyridyl Cephalosporins of Formula I:
Compounds 505, 506, and 507
Figure imgf000137_0001
22 23
A. Preparation of 2-bromo-5-(((tert-butyldimethylsilyl)oxy)methyl)pyridine 23
[0309] Compound 23 is prepared by a literature procedure (U.S. Patent 5,283,242). More specifically, under anhydrous conditions, (6-bromopyridin-3- yl)methanol 22 (from Combi-Blocks) (1.000 g, 5.319 mmol, 1.000 equiv) is dissolved in anhydrous DMF (8 ml_, 0.7 M) at rt. At rt, triethylamine (0.771 ml_, 5.53 mmol, 1.04 equiv) is added slowly. At 0 °C, fe/f-butyldimethylsilyl chloride (0.834 g, 5.53 mmol, 1.04 equiv) is added as a solid and the remaining material in the weighing vial is transferred as a solution in DMF (2 ml_). The cloudy suspension is stirred at rt for 1 hour and 10 minutes. Diethyl ether (50 ml_) is added as well as water (50 ml_). Layers were separated and the organic layer is washed with water (40 mL), dried over magnesium sulfate and filtered. Solvent is evaporated under reduced pressure. The yellow oil obtained is chromatographed on silica gel (Biotage SNAP, 25 g), eluting with 100% DCM. The desired product 23 is isolated as colorless oil (1.35 g, 4.47 mmol, 84% yield). [0310] 1H NMR (500 MHz, CDCb) d (ppm) 8.32 (d, 1 H , J = 2.5 Hz), 7.54 (dd, 1 H, J = 8.5 Hz, J = 2.5 Hz), 7.46 (d, 1 H, J = 8.5 Hz), 4.72 (s, 2H), 0.94 (s, 9H), 0.1 1 (s, 6H). m/z = 302, 304 [M+H]+, 343, 345 [M+H+ACN]+.
Figure imgf000138_0001
B. Preparation of 5-(((tert-butyldimethylsilyl)oxy)methyl)-2-(tributylstannyl)pyridine 24
[0311] Under anhydrous conditions, substrate 23 (1 .35 g, 4.47 mmol, 1 .00 equiv) is dissolved in anhydrous THF (1 1 mL, 0.40 M). At - 78 °C, a 2.5 M solution of n-BuLi in hexanes (1 .88 mL, 4.69 mmol, 1 .05 equiv) is added dropwise. The solution turns orange and then dark green. The solution is then stirred at - 78 °C for 1 hour. At - 78 °C, a solution of tri-n-butyltin chloride (from Combi-Blocks) (2.98 g, 9.15 mmol, 2.05 equiv) in anhydrous THF (2.3 mL, 4 M) is slowly added. The dark green solution turn yellow; it is then stirred at - 78 °C for 1 .5 hours. The reaction is quenched by slow addition of water (10 mL) at rt. Diethyl ether (50 mL) is added followed by more water (50 mL). Layers are separated and organic layer is washed with water (50 mL), dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. Obtained yellow oil is chromatographed on silica gel (Biotage SNAP, 100 g), eluting with 0-20% AcOEt/Hexanes. The desired product 24 is obtained as yellow oil (1 .42 g, 2.78 mmol, 62% yield). It is contaminated by some tin by-product, but is used without further purification in the next step.
[0312] 1H NMR (500 MHz, CDCb) d (ppm) 8.71 (d, 1 H, J = 1.5 Hz), 7.48 (dd, 1 H), 7.39 (d, 1 H, J = 8.0 Hz), 4.73 (s, 2H), 1 .70-1.63 (m, 6H), 1 .40-1 .27 (m, 12H), 0.94 (s, 9H), 0.92-0.85 (m, 9H), 0.1 1 (s, 6H). m/z = 514.2 [M+H]+.
Figure imgf000139_0001
C. Preparation of (6R,7R)-benzhvdryl 3-(5-(((tert-butyldimethylsilyl)-
Figure imgf000139_0002
oxy)methyl)pyridin-2-yl)-8-oxo-7-(2-phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2- ene-2-carboxylate 25
[0313] Preparation of compound 25 is adapted from U.S. Patent 4,870, 168. Under anhydrous conditions, the triflate 4 (5.04 g, 7.97 mmol, 1 .00 equiv) is dissolved in anhydrous NMP (50 ml_, 0.16 M). To this orange suspension, the stannane 24 (5.51 g, 10.8 mmol, 1 .35 equiv) is added at rt as a solution in anhydrous NMP (10 ml_, 1 .1 M). At rt, zinc chloride (2.17 g, 15.9 mmol, 2.00 equiv) and tri(2-furyl)phosphine (1.48 g, 6.37 mmol, 0.80 equiv) are added to the mixture. Nitrogen gas is bubbled into the solution for five minutes. Then, at rt, palladium bis(dibenzylidene acetone) (1 .83 g, 1 .19 mmol, 0.40 equiv) is added in one portion. The deep purple/dark brown mixture is immersed in an oil bath, preheated to 55 °C. The heated mixture is stirred at 55 °C for 19 hours. Heating is stopped and the mixture is diluted in ethyl acetate (250 ml_) and is washed with saturated ammonium chloride solution (150 + 100 ml_). Combined aqueous layers are extracted using ethyl acetate (150 ml_). The combined organic layers are washed with brine and solvent is evaporated under reduced pressure. The residue (dark brown oil) is dissolved in acetonitrile (150 ml_) and this layer is washed with hexanes (2 x 80 ml_). Solvent is evaporated under reduced pressure. The obtained dark brown oil is purified by column chromatography on silica gel (Biotage SNAP, 100 g) eluting with 0-100% AcOEt/Hexanes. The obtained product is purified by a second column chromatography on silica gel (Biotage SNAP, 50 g) eluting with 0-80% AcOEt/Hexanes. The desired product 25 is obtained as red solid, contaminated by some by-product (1 .29 g). The product is used without further purification in the next step.
[0314] 1H NMR (500 MHz, CDCh) d (ppm) 8.29 (s, 1 H), 7.45-7.15 (m, 13H), 7.05- 6.99 (m, 2H), 6.89 (s, 1 H), 6.88-6.83 (m, 2H), 6.15-6.05 (m, 1 H), 5.95-5.89 (m, 1 H), 5.06 (d, 1 H, J = 5.0 Hz), 4.58 (s, 2H), 4.14 (d, 1 H, J = 19.0 Hz), 3.70-3.60 (m, 2H), 3.53 (d, 1 H, J = 18.5 Hz), 0.95 (s, 9H), 0.1 1 (s, 6H). m/z = 706 [M+H]+.
Figure imgf000140_0001
D. Preparation of (6R,7R)-benzhvdryl 3-(5-(hvdroxymethyl)pyridin-2-yl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 26
[0315] Under inert atmosphere, TBS-protected substrate 25 (not pure, 1 .069 g, 1 .514 mmol, 1 .000 equiv) is dissolved in anhydrous THF (5.0 ml_, 0.30 M) at rt. This solution is cooled to 0 °C and acetic acid (0.87 ml_, 15 mmol, 10 equiv) is added. At 0 °C, the 1 .0 M TBAF solution in TFIF (4.54 ml_, 4.54 mmol, 3.00 equiv) is added dropwise. The resulting solution is stirred at 0 °C for 3 hours and 10 minutes. AcOEt (50 ml_) and sat. aqueous NaFICOs (40 ml_) are added. Layers are separated and the organic layer is washed with brine and dried over sodium sulfate. Solvent is evaporated under reduced pressure. The orange film obtained is chromatographed on silica gel (Biotage SNAP, 25 g), eluting with 20-100% AcOEt/Hexanes. The desired compound 26 is obtained as pale orange solid (0.384 g, 0.649 mmol, 8% yield over 2 steps).
[0316] 1H NMR (500 MHz, CDCL) d (ppm) 8.30 (s, 1 H), 7.44-7.37 (m, 2H), 7.35- 7.17 (m, 1 1 H), 7.09-7.03 (m, 2H), 6.97-6.92 (m, 2H), 6.88 (s, 1 H), 6.05 (d, 1 H, J = 9.0 Hz), 5.95 (dd, 1 H, J = 9.5 Hz, J = 5.0 Hz), 5.08 (d, 1 H, J = 5.0 Hz), 4.55 (br s, 2H), 4.14 (d, 1 H, J = 18.5 Hz), 3.71 (d, 1 H, J = 16.0 Hz), 3.65 (d, 1 H, J = 16.0 Hz), 3.56 (d, 1 H, J = 19.0 Hz). m/z = 592 [M+H]+.
Figure imgf000140_0002
E. Preparation of (6R,7R)-benzhvdryl 3-(5-(chloromethyl)pyridin-2-yl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 27
[0317] Under anhydrous conditions, compound 26 (0.410 g, 0.693 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (6.9 ml_, 0.10 M). At 0 °C, methanesulfonyl chloride (0.080 ml_, 1 .0 mmol, 1 .5 equiv) is added dropwise, followed by dropwise addition of /V-methylmorpholine (0.080 ml_, 0.73 mmol, 1.05 equiv). The orange solution is stirred at 0 °C for 50 minutes and then at rt for 1 hour and 40 minutes. MsCI (0.016 ml_, 0.30 equiv) is then added. The mixture is stirred at rt for 0.5 hour. Additional MsCI (0.016 ml_, 0.30 equiv) is added and the solution is stirred at rt for another 0.5 hour. Solvent is evaporated to dryness. Lithium chloride (0.059 g, 1 .4 mmol, 2.0 equiv) is added to the residue and anhydrous DMF (6.9 mL, 0.10 M) is added. The resulting orange solution is stirred at rt for 1 hour and is then diluted in ethyl acetate (50 mL) and sat. aqueous NhUCI (50 mL) is added. Layers were separated and the organic layer is washed once more with sat. aqueous NhUCI (20 mL). The aqueous washes are combined and extracted using AcOEt (30 mL). The organic extracts are combined, washed with brine, dried over magnesium sulfate and filtered. Solvent is evaporated under reduced pressure. The brown solid obtained is chromatographed on silica gel (Biotage SNAP, 25 g) eluting with 10-100% AcOEt/Hexanes gradient. The desired product 27 is obtained as pale orange solid (0.402 g, 0.659 mmol, 95% yield). This compound is contaminated by ca. 20% of what is believed to be a double-bond migration isomer. However, the product is used without further purification in the next step.
[0318] 1H NMR (500 MHz, CDCh) d (ppm) 8.30 (s, 1 H), 7.42-7.20 (m, 14H), 7.08- 7.02 (m, 2H), 6.92-6.87 (m, 2H), 6.06 (d, 1 H, J = 9.5 Hz), 5.96 (dd, 1 H, J = 9.0 Hz, J = 4.5 Hz), 5.08 (d, 1 H, J = 5.0 Hz), 4.42 (d, 1 H, J = 12.0 Hz), 4.40 (d, 1 H, J = 12.5 Hz), 4.14 (d, 1 H, J = 19.0 Hz), 3.72 (d, 1 H, J = 16.0 Hz), 3.65 (d, 1 H, J = 16.0 Hz), 3.55 (d, 1 H, J = 18.5 Hz) m/z = 609.6 [M+H]+.
Figure imgf000142_0001
F. Preparation of (6R,7R)-benzhvdryl 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-3- (5-(chloromethyl)pyridin-2-yl)-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 29
[0319] Under anhydrous conditions, phosphorus pentachloride (0.837 g, 4.02 mmol, 6.10 equiv) is suspended in anhydrous DCM (6.6 mL, 0.10 M). At - 10 °C, pyridine (0.325 mL, 4.02 mmol, 6.10 equiv) is added dropwise. To the obtained white suspension, the chloride substrate 27 (0.402 g, 0.659 mmol, 1.00 equiv) is added dropwise as a solution in anhydrous DCM (4.4 mL, 0.15 M), at - 10 °C. The resulting pale-yellow suspension is stirred between - 10 °C and 0 °C for 1 hour and 15 minutes. At 0 °C, anhydrous methanol (2.67 mL, 65.9 mmol, 100 equiv) is added dropwise and the solution is stirred between 0 °C and 10 °C for 20 minutes. This reaction mixture is diluted in DCM (50 mL) and saturated sodium bicarbonate solution (40 mL) is carefully added. Layers are separated and the aqueous wash is extracted using DCM (30 mL). The combined organic extracts are washed with sat. sodium bicarbonate solution (40 mL), dried over magnesium sulfate, filtered and solvent is evaporated until ca. 10 of solution remained. [0320] In parallel, a solution of acid chloride is prepared as follows:
[0321] Under anhydrous conditions, phosphorus pentachloride (0.189 g, 0.906 mmol, 1 .20 equiv) is suspended in anhydrous DCM (3.8 mL, 0.20 M) at rt. The resulting white suspension is stirred at 0 °C for 10 minutes. At 0 °C, the carboxylic acid 8 (0.324 g, 0.755 mmol, 1 .00 equiv.) is added in one portion. The heterogeneous mixture of acid chloride is stirred, between at 0 °C for 1 hour.
[0322] To the acid chloride 9 solution is added the free amine 28 solution (ca. 10 ml_) dropwise at 0 °C. The resulting brown solution is stirred between 0 °C and rt over 1 hour. This reaction mixture is then diluted in DCM (50 ml_), and washed with water (40 + 50 ml_). The organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The obtained orange film is purified by normal phase chromatography (Biotage SNAP, 25 g) eluting with 10-80% AcOEt/Hexanes. Fractions containing the desired product are combined and evaporated under reduced pressure. This material is further purified by normal phase chromatography on silica gel (Biotage SNAP, 25 g), eluting with 10-80% AcOEt/Hexanes. The desired product 29 is separated from its double-bond migration isomer and it is obtained as off-white solid (0.335 g, 0.371 mmol, 56% yield).
[0323] 1H NMR (500 MHz, CDCIs) d (ppm) 8.32 (s, 1 H), 8.10 (d, 1 H, J = 9.0 Hz), 7.35-7.17 (m, 9H), 7.10-7.04 (m, 2H), 6.95-6.88 (m, 2H), 6.14 (dd, 1 H, J = 8.5 Hz, J = 5.5 Hz), 5.19 (d, 1 H, J = 4.5 Hz), 4.44 (d, 1 H, J = 12.5 Hz), 4.40 (d, 1 H, J = 1 1 .5 Hz), 4.23 (d, 1 H, J = 18.5 Hz), 3.60 (d, 1 H, J = 18.5 Hz), 1 .66 (s, 3H), 1 .62 (s, 3H), 1 .54 (s, 9H), 1 .42 (s, 9H). m/z = 902.2 [M]+.
Figure imgf000143_0001
G. Preparation of (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-(5-(chloromethyl)pyridin-2-yl)-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-ene-2-carboxylic acid 30 (505)
[0324] Under nitrogen atmosphere, substrate 29 (0.335 g, 0.371 mmol, 1.00 equiv) is dissolved in anhydrous DCM (3.7 ml_, 0.10 M). At 0 °C, anisole (0.201 ml_, 1 .85 mmol, 5.00 equiv) is added slowly, quickly followed by slow addition of TFA (5.3 ml_, 0.070 M). The pale-yellow solution is stirred at 0 °C for 2 hours and then at rt for 1 .5 hours. This reaction mixture is then evaporated to dryness. The orange oil obtained is triturated in diethyl ether (4 x 4 ml_). Remaining solvent is evaporated and traces of diethyl ether are removed by co-evaporation with hexanes (5 ml_). The desired product 30 is obtained as pale purple solid (0.220 g, quantitative yield), which is used without further purification in the next step.
[0325] 1H NMR (300 MHz, Acetone-de) d (ppm) 8.64 (d, 1 H, J = 1.5 Hz), 7.86 (dd, 1 H, J = 8.1 Hz, J = 2.4 Hz), 7.44 (d, 1 H, J = 8.1 Hz), 7.05 (s, 1 H), 6.08 (d, 1 H, J = 4.8 Hz), 5.34 (d, 1 H, J = 4.8 Hz), 4.79 (s, 2H), 4.26 (d, 1 H, J = 18.3 Hz), 3.76 (d, 1 H, J = 18.3 Hz), 1 .65 (s, 3H), 1 .61 (s, 3H). m/z = 581 .2 [M+H]+.
Figure imgf000144_0001
H. Preparation of 2-((6-((6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-en-3-yl)pyridin-3-yl)methyl)-6,7-dihvdroxyisoquinolin-2-ium 31 (506)
[0326] Under nitrogen atmosphere, 6,7-dihydroxyisoquinolin-2-ium bromide 12 (0.033 g, 0.14 mmol, 8.0 equiv), potassium carbonate (0.014 g, 0.10 mmol, 6.0 equiv) and substrate 30 (0.010 g, 0.017 mmol, 1 .0 equiv) were suspended in anhydrous DMF (0.17 ml_, 0.10 M). To the brown suspension, containing white solids, potassium iodide (0.006 g, 0.03 mmol, 2 equiv) is added. The reaction mixture is stirred at rt, under nitrogen atmosphere for 3 hours and 40 minutes. TFA (3 drops) is then added, followed by ca. 0.3 ml_ of water. The resulting solution is chromatographed on a reverse phase column (Teledyne GOLD C18, 15.5 g), eluting with a 0-100% ACN/H2O gradient (+ 0.1 % TFA). Fractions containing the desired product are combined and lyophilized. The desired product 31 is obtained as pale purple solid (0.009 g, 0.013 mmol, 74% yield).
[0327] 1H NMR (300 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 9.66 (s, 1 H),
Figure imgf000145_0001
I. Preparation of 1 -((6-((6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-en-3-yl)pyridin-3-yl)methyl)-5.6-dihvdroxy-2-methyl-2H-indazol-
1 -ium 32 (507)
[0328] Under nitrogen atmosphere, 2-methyl-2/-/-indazol-1 -ium bromide 14 (0.050 g, 0.21 mmol, 12 equiv), potassium carbonate (0.019 g, 0.14 mmol, 8.0 equiv) and substrate 30 (0.010 g, 0.017 mmol, 1 .0 equiv) are suspended in anhydrous DMF (0.2 ml_, 0.1 M). Potassium iodide (0.006 g, 0.03 mmol, 2 equiv) is added and the resulting cloudy purple mixture is immersed in a preheated oil bath (40 °C) and is stirred at 40 °C for 1 .5 hours. The heating is stopped and 3 drops of TFA are added, followed by ca. 0.5 ml_ of water. This solution is charged onto a reverse phase column (Biotage C18, 15.5 g) and is eluted with a 5-195% ACN/FI2O (+ 0.1 % TFA) gradient. Fractions containing the desired product were combined and lyophilized. The desired product 32 is obtained as white solid (0.002 g, 0.003 mmol, 16% yield).
[0329] 1H NMR (300 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 8.84 (s, 1 H), 8.52 (s, 1 H), 7.61 (dd, 1 H, J = 8.1 Hz, J = 2.7 Hz), 7.43 (d, 1 H, J = 8.4 Hz), 7.30 (s, 1 H), 7.26 (s, 1 H), 7.04 (s, 1 H), 6.1 1 (s, 2H), 6.04 (d, 1 H, J = 4.2 Hz), 5.31 (d, 1 H, J = 4.8 Hz), 4.38 (s, 3H), 4.18 (d, 1 H, J = 18.0 Hz), 3.73 (d, 1 H, J = 18.9 Hz), 1 .62 (s, 3H), 1 .59 (s, 3H). m/z = 709.4 [M]+, 355.2 [M+H]2+. Example 3: Preparation of Exemplary 2-Pyridyl Cephalosporins of Formula I: Compounds 512, 513
Figure imgf000146_0001
33
34
A. Preparation of 5-bromo-2-(((fe/f-butyldimethylsilyl)oxy)methyl)pyridine 34
[0330] Compound 34 is prepared by a literature procedure (WO 2017/16261 1 , PCT/EP2017/056599). More specifically, under anhydrous conditions, (5- bromopyridin-2-yl)methanol 33 (from Combi-Blocks) (2.000 g, 10.64 mmol, 1.000 equiv) is dissolved in anhydrous DMF (19 mL, 0.56 M) at rt. At rt, imidazole (1 .09 g, 16.0 mmol, 1 .50 equiv) is added in one portion. At 0 °C, fe/f-butyldimethylsilyl chloride (1.92 g, 12.8 mmol, 1 .20 equiv) is added as a solid and the remaining material in the weighing vial is transferred as a solution in DMF (2 mL). The resulting clear yellow solution is stirred at rt for 1 hour and 45 minutes. AcOEt (70 mL) and water (70 mL) are then added. Layers are separated and the organic layer is washed with water (40 mL), then with brine. It is then dried over sodium sulfate and filtered. Solvent is evaporated under reduced pressure. The pale-yellow oil obtained is chromatographed on silica gel (Biotage SNAP, 100 g), eluting with 100% DCM. The desired product 34 is isolated as colorless oil (3.17 g, 10.5 mmol, 99% yield).
[0331] 1H NMR (500 MHz, CDC ) d (ppm) 8.57 (d, 1 H, J = 2.5 Hz), 7.83 (dd, 1 H, J = 8.5 Hz, J = 2.0 Hz), 7.43 (d, 1 H, J = 7.5 Hz), 4.78 (s, 2H), 0.96 (s, 9H), 0.13 (s, 6H). m/z = 302, 304 [M+H]+.
Figure imgf000146_0002
B. Preparation of 2-(((fe/f-butyldimethylsilyl)oxy)methyl)-5-(tributylstannyl)pyridine 35
[0332] Under anhydrous conditions, substrate 34 (1 .50 g, 4.96 mmol, 1 .00 equiv) is dissolved in anhydrous THF (12 ml_, 0.40 M). At - 78 °C, a 2.5 M solution of n-BuLi in hexanes (2.08 ml_, 5.21 mmol, 1 .05 equiv) is added dropwise. The solution turned to orange and then to dark brown. The solution is stirred at - 78 °C for 40 minutes. At - 78 °C, a solution of tri-n-butyltin chloride (from Combi-Blocks) (1 .41 g, 5.21 mmol, 1 .05 equiv) in anhydrous THF (1 .3 ml_, 4 M) is slowly added. The dark brown solution turned to orange; it is then stirred between - 78 °C and rt for 3 hours. The reaction is quenched by slow addition of water (12 ml_) at rt. Diethyl ether (50 ml_) is added followed by more water (40 ml_). Layers are separated and the organic layer is washed with water (50 mL), dried over sodium sulfate and filtered. Solvent is evaporated under reduced pressure. The orange oil obtained is diluted in ACN (40 mL) and this organic solution is extracted with hexanes (40 mL). The hexanes layer is filtered through celite and solvent is evaporated. The desired compound 35 is obtained as orange oil (2.44 g, 4.76 mmol, 96% yield). The product is contaminated by some tin by-product, but is used without further purification in the next step.
[0333] 1H NMR (500 MHz, CDCL) d (ppm) 8.49 (s, 1 H), 7.76 (d, 1 H, J = 8.5 Hz), 7.46 (d, 1 H, J = 8.0 Hz), 4.82 (s, 2H), 1 .58-1.50 (m, 6H), 1.42 -1.32 (m, 12H), 1 .15-1 .05 (m, 6H), 0.97 (s, 9H), 0.95-0.85 (m, 9H), 0.13 (s, 9H).
Figure imgf000147_0001
C. Preparation of (6R,7R)-benzhvdryl 3-chloro-8-oxo-7-(2-phenylacetamido)-5- thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 36
[0334] Under anhydrous conditions, (6R,7R)-benzhydryl 3-hydroxy-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate 3 (from AK Scientific) (5.00 g, 9.99 mmol, 1 .00 equiv) is dissolved in anhydrous DMF (30 mL, 0.33 M). To the pale-brown solution, at rt, phosphorus oxychloride (1 .74 mL, 9.99 mmol, 1 .00 equiv) is added dropwise. This solution is stirred at rt for 4 hours and becomes dark brown over time. The reaction solution is left at 4 °C, without stirring for 3 days. The reaction solution is then transferred to a larger flask and water (100 ml_) is gradually added to the dark brown solution. Orange precipitate appears while a small exotherm is observed. The precipitate is collected by filtration on sintered glass. The orange precipitate is washed with water (3 x 100 ml_) and dried on a sintered glass funnel. The solid (hydrated) is dried under high vacuum for 24 hours. Product 36 is obtained as pale yellow solid (5.08 g, 9.79 mmol, 98% yield). This product is used without further purification in the next step.
[0335] 1H NMR (500 MHz, CDC ) d (ppm) 7.43-7.20 (m, 15H), 6.99 (s, 1 H), 5.99 (d, 1 H, J = 9.5 Hz), 5.83 (dd, 1 H, J = 9.5 Hz, J = 5.0 Hz), 5.02 (d, 1 H, J = 5.0 Hz), 3.77 (d, 1 H, J = 18.5 Hz), 3.68 (d, 1 H, J = 16.5 Hz), 3.62 (d, 1 H, J = 16.0 Hz), 3.45 (d, 1 H, J = 18.5 Hz).
Figure imgf000148_0001
D. Preparation of (6R,7R)-benzhvdryl 3-(6-(((tert-butyldimethylsilyl)oxy)methyl)- pyridin-3-yl)-8-oxo-7-(2-phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2- carboxylate 37
[0336] Under anhydrous conditions, the chloride 36 (1 .00 g, 1 .93 mmol, 1 .00 equiv) is introduced into a flask. A solution of stannane 35 (1 09g, 2.12 mmol, 1 .10 equiv) in anhydrous NMP (14 mL, 0.15 M) is added and nitrogen is bubbled into the resulting orange solution. At rt, zinc chloride (0.53 g, 3.9 mmol, 2.0 equiv) is added, followed by tri(2-furyl)phosphine (0.179 g, 0.771 mmol, 0.40 equiv), while nitrogen is still being bubbled into the solution. At rt, tris(dibenzylideneacetone)dipalladium (0.353 g, 0.385 mmol, 0.20 equiv) is added in one portion. The resulting deep green/brown mixture is immersed in a preheated oil bath (70 °C) and is stirred at 70 ° for 1 .5 hours. Heating is then stopped, and the reaction mixture is diluted in AcOEt (60 mL). This organic layer is washed with sat. ammonium chloride solution (3 x 50 mL) and then with brine. Solvent is evaporated under reduced pressure. The resulting brown oily film is taken up in ACN (50 mL). This organic layer is washed with hexanes (2 x 50 mL), dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is chromatographed on silica gel (Biotage SNAP, 50 g) eluting with 0-60% AcOEt/Hexanes. The desired compound 37 is obtained as an orange solid (0.769 g, 1 .09 mmol, 57% yield). The product is used without further purification in the next step.
[0337] 1H NMR (500 MHz, CDCIs) d (ppm) 8.25 (s, 1 H), 7.45-7.38 (m, 3H), 7.37- 7.22 (m, 8H), 7.23-7.14 (m, 4H), 6.98-6.93 (m, 2H), 6.81 (s, 1 H), 6.04 (d, 1 H, J = 9.0 Hz), 5.93 (dd, 1 H, J = 9.0 Hz, J = 5.0 Hz), 5.07 (d, 1 H, J = 5.0 Hz), 4.75 (d, 1 H, J = 15.5 Hz), 4.71 (d, 1 H, J = 15.5 Hz), 3.72 (d, 1 H, J = 16.0Hz), 3.65 (d, 1 H, J = 16.5 Hz), 3.61 (d, 1 H, J = 18.5 Hz), 3.52 (d, 1 H, J = 18.5 Hz), 0.98 (s, 9H), 0.14 (s, 6H). m/z = 706.0 [M+H]+.
Figure imgf000149_0001
E. Preparation of (6R,7R)-benzhvdryl 3-(6-(hvdroxymethyl)pyridin-3-yl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 38
[0338] Under inert atmosphere, TBS-protected substrate 37 from step D (0.765 g, 1 .08 mmol, 1 .00 equiv) is dissolved in anhydrous THF (3.1 mL, 0.35 M) at rt. This solution is cooled to 0 °C and acetic acid (0.62 mL, 1 1 mmol, 10 equiv) is added. At 0 °C, a 1 .0 M TBAF solution in THF (1 .84 mL, 1 .84 mmol, 1 .70 equiv) is added dropwise. The resulting solution is stirred at 0 °C for 5 hours and 10 minutes. AcOEt (50 mL) and sat. aqueous NaHCCb (50 mL) are added. Layers are separated and the organic layer is washed with brine and dried over sodium sulfate. Solvent is evaporated under reduced pressure. The brown solid obtained is dried under high vacuum. The desired product 38 is obtained as brown solid (no yield derived). This compound is used without further purification in the next step.
[0339] 1H NMR (500 MHz, CDCIs) d (ppm) 8.28 (s, 1 H), 7.77-7.73 (m, 1 H), 7.45- 7.15 (m, 13H), 7.04-6.96 (m, 3H), 6.81 (s, 1 H), 6,08-6.03 (m, 1 H), 5.97-5.93 (m, 1 H), 5.09 (d, 1 H, J = 5.0 Hz), 4.66 (s, 2H), 3.73 (d, 1 H, J = 16.5 Hz), 3.66 (d, 1 H, J = 16.5 Hz), 3.63 (d, 1 H, J = 18.5 Hz), 3.53 (d, 1 H, J = 18.5 Hz) m/z = 592.2 [M+H]+.
Figure imgf000150_0001
F. Preparation of (6F?,7F?)-benzhvdryl 3-(6-(chloromethyl)pyridin-3-yl)-8-oxo-7-(2- phenylacetamido)-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 39
[0340] Under anhydrous conditions, compound 38 (0.641 g, 1.08 mmol, 1.00 equiv) is dissolved in anhydrous DMF (5.4 ml_, 0.20 M). At rt, phosphorus oxychloride (0.152 ml_, 1.63 mmol, 1.50 equiv) is added dropwise. The resulting dark orange solution is stirred at rt for 1.5 hours. The reaction mixture is diluted in AcOEt (60 ml_) and this organic layer is washed with water (60 ml_), sat. ammonium chloride solution (50 ml_) and brine. The combined aqueous washes are extracted using AcOEt (40 ml_). The organic extracts are combined, dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The resulting dark orange film is chromatographed on silica gel (Biotage SNAP, 25 g) eluting with 0-50% AcOEt/Hexanes. Fractions containing the desired product are combined and solvent is evaporated under reduced pressure. Orange solid is obtained, consisting of the desired product 39 (0.398 g, 0.652 mmol, 60% yield), contaminated with about 10% of the double-bond migration isomer. The orange solid is used without further purification in the next step.
[0341] 1H NMR (500 MHz, CDCIs) d (ppm) 8.31 (d, 1 H, J = 1.5 Hz), 7.75 (s, 1 H), 7.45-7.15 (m, 14H), 7.03-6.98 (m, 2H), 6.80 (s, 1 H), 6.03 (d, 1 H, J = 8.5 Hz), 5.96 (dd,
1 H, J = 8.5 Hz, J = 4.5 Hz), 5.08 (d, 1 H, J = 5.0 Hz), 4.56 (s, 2H), 3.72 (d, 1 H, J = 17.0 Hz), 3.65 (d, 1 H, J = 17.0 Hz), 3.62 (d, 1 H, J = 19.0 Hz), 3.52 (d, 1 H, J = 19.5 Hz) m/z = 610.1 [M+H]+.
Figure imgf000151_0001
G. Preparation of (6R,7R)-benzhvdryl 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-3- (6-(chloromethyl)pyridin-3-yl)-8-oxo-5-thia-1 -azabicvclo[4.2.01oct-2-ene-2-carboxylate 41
[0342] Under anhydrous conditions, phosphorus pentachloride (0.425 g, 3.48 mmol, 5.00 equiv) is suspended in anhydrous DCM (4.6 ml_, 0.15 M). At - 10 °C, and pyridine (0.282 ml_, 3.48 mmol, 5.00 equiv) is added dropwise. The resulting white suspension is stirred at rt for 15 minutes. Then, at - 10 °C, the chloride substrate 39 (0.425 g, 0.697 mmol, 1 .00 equiv) is added dropwise as a solution in anhydrous DCM (3.5 ml_, 0.20 M). The resulting orange suspension is stirred between - 10 °C and 0 °C for 1 hour and 15 minutes. At 0 °C, anhydrous methanol (2.82 ml_, 69.7 mmol, 100 equiv) is added dropwise and the solution is stirred between 0 °C and 10 °C for 20 minutes. This reaction mixture is diluted in DCM (20 ml_) and saturated sodium bicarbonate solution (20 ml_) is carefully added. Layers are separated and the aqueous wash is extracted with DCM (15 mL). The combined organic extracts are washed with sat. sodium bicarbonate solution (20 mL), water (20 mL) and brine. The washed organic extracts are dried over sodium sulfate, filtered and solvent is evaporated until ca. 5 to 10 mL of solution remains. In parallel, a solution of acid chloride 9 is prepared as follows:
[0343] Under anhydrous conditions, phosphorus pentachloride (0.192 g, 0.921 mmol, 1.15 equiv) is suspended in anhydrous DCM (3.2 mL, 0.25 M) at rt. The resulting white suspension is stirred at 0 °C for 15 minutes. At 0 °C, the carboxylic acid 8 (0.344 g, 0.801 mmol, 1.00 equiv) is added in one portion. The heterogeneous mixture is stirred, at 0 °C for 20 minutes.
[0344] To the latter acid chloride 9 solution is added the free amine 39 solution (ca. 5-10 mL) dropwise at 0 °C. The resulting orange solution is stirred between 0 °C and rt for 1 hour and 10 minutes. This reaction mixture is then diluted in DCM (40 mL), and washed with water (40 mL). The organic layer is washed with water (2 x 40 mL), then with brine. The washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The orange film, which is obtained, is purified by normal phase chromatography (Biotage SNAP, 25 g), eluting with 0-80% AcOEt/Hexanes. Fractions containing the desired product are combined and evaporated under reduced pressure. The desired product 29 could be separated from its double-bond migration isomer and it is obtained as pale orange solid (0.386 g, 0.427 mmol, 61% yield).
[0345] 1H NMR (500 MHz, CDCIs) d (ppm) 8.34 (s, 1 H), 8.22 (br s, 1 H), 7.40-7.32 (m, 2H), 7.37-7.15 (m, 10H), 70.6-7.02 (m, 2H), 7.04 (s, 1 H), 6.12 (dd, 1 H, J = 9.0 Hz, J = 5.5 Hz), 5.02 (d, 1 H, J = 5.0 Hz), 4.57 (s, 2H), 3.67 (d, 1 H, J = 18.5 Hz), 3.60 (d, 1 H, J = 18.5 Hz), 1.68 (s, 3H), 1.64 (s, 3H), 1.55 (s, 9H), 1.44 (s, 9H). m/z = 903 [M+H]+.
Figure imgf000152_0001
H. Preparation of (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-(6-(chloromethyl)pyridin-3-yl)-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-ene-2-carboxylic acid 42 (512)
[0346] Under nitrogen atmosphere, substrate 41 (0.380 g, 0.421 mmol, 1.00 equiv) is dissolved in anhydrous DCM (4.2 ml_, 0.10 M). At 0 °C, anisole (0.18 ml_, 1 .7 mmol, 4.0 equiv) is added slowly, quickly followed by slow addition of TFA (5.3 ml_, 0.080 M). The yellow solution is stirred at 0 °C for 3 hours and 40 minutes and then at rt for 1 hour and 20 minutes. This reaction mixture is then evaporated to dryness. The orange oil obtained is triturated in diethyl ether (3 x 4 ml_). Remaining solvent is evaporated and traces of diethyl ether are removed by co-evaporation with hexanes (2 x 5 ml_). The desired product 42 is obtained as off-white solid (0.272 g, quantitative yield), which is used without further purification in the next step.
[0347] 1H NMR (500 MHz, Acetone-de) d (ppm) 8.58 (d, 1 H, J = 8.5Hz), 8.47 (d,
1 H, J = 2.5 Hz), 7.84 (dd, 1 H, J = 7.5 Hz, J = 2.5 Hz), 7.58 (d, 1 H, J = 8.0 Hz), 7.02 (s, 1 H), 6.12 (dd, 1 H, J = 9.5 Hz, J = 5.0 Hz), 5.37 (d, 1 H, J = 5.0 Hz), 4.74 (s, 2H), 3.91 (s, 2H), 1 .69 (s, 3H), 1 .64 (s, 3H). m/z = 581 [M+H]+.
Figure imgf000153_0001
I. Preparation of 2-((5-((6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 - azabicvclo[4.2.01oct-2-en-3-yl)pyridin-2-yl)methyl)-6,7-dihvdroxyisoquinolin-2-ium 43 (513)
[0348] Under nitrogen atmosphere, 6,7-dihydroxyisoquinolin-2-ium bromide 12 (0.996 g, 4.12 mmol, 10.0 equiv), potassium carbonate (0.367 g, 2.68 mmol, 6.5 equiv) and substrate 42 (0.240 g, 0.412 mmol, 1 .0 equiv) are suspended in anhydrous DMF (2.9 ml_, 0.14 M). To the orange suspension, containing white solids, potassium iodide (0.205 g, 1 .24 mmol, 3.00 equiv) is added. The reaction mixture is stirred at rt, under nitrogen atmosphere for 2 hours. Potassium iodide (0.068 g, 0.412 mmol, 1 .00 equiv) is added and the suspension is stirred at rt for 1 hour and 50 minutes. TFA (1 .5 ml_, 20 mmol, 48 equiv) is then added, followed by ca. 2.5 ml_ of water. The resulting solution is chromatographed on a reverse phase column (Teledyne GOLD C18, 60 + 60 g), eluting with a 0-100% ACN/H2O gradient (+ 0.1 % TFA). Fractions containing the desired product are combined and lyophilized. The desired product 43 is obtained as off-white solid (0.249 g, 0.352 mmol, 86% yield).
[0349] 1H NMR (300 MHz, Acetone-de + D2O (+1 % TFA)) d (ppm) 9.57 (s, 1 H), 8.43 (dd, 1 H, J = 6.9 Hz, J = 0.9 Hz), 8.40 (d, 1 H, J = 2.1 Hz), 8.13 (d, 1 H, J = 6.9 Hz), 7.89 (dd, 1 H, J = 7.8 Hz, J = 2.1 Hz), 7.74 (s, 1 H), 7.70 (d, 1 H, J = 8.1 Hz), 7.56 (s, 1 H), 6.99 (s, 1 H), 6.09-6.00 (m, 3H), 5.31 (d, 1 H, J = 4.8 Hz), 3.82 (s, 2H), 1 .61 (s, 3H), 1 .58 (s, 3H). m/z = 705.8 [M]+, 353.4 [M+H]2+.
Example 4: Preparation of Exemplary Monobactams of Formula I: Compounds 476 477 478 479 and 481
Figure imgf000154_0001
A. Preparation of 4-(((tert-butyldimethylsilyl)oxy)methyl)aniline 45
[0350] (4-aminophenyl)methanol 44 (1 .00 g, 8.13 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (20 mL, 0.40 M). At rt, fe/f-butyldimethylsilyl chloride (1 .47 g, 9.76 mmol, 1 .20 equiv) is added and the resulting solution is stirred at rt for 5 minutes. Triethylamine (2.3 mL, 16 mmol, 2.0 equiv) is added at rt and the reaction mixture is stirred at rt for 4 hours. The reaction mixture is diluted in AcOEt and this organic layer is washed with water, sat. aqueous NH4CI, sat. aqueous NaHCCb and then with brine. The organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The resulting colourless oil is purified by column chromatography on silica gel. The desired product 45 is obtained as a colourless oil (1 .20 g, 5.06 mmol, 62% yield).
[0351] 1H NMR (500 MHz, CDCIs) d (ppm) 7.15-7.10 (m, 2H), 6.69-6.64 (m, 2H), 4.63 (s, 2H), 3.62 ( br s, 2H), 0.93 (s, 9H), 0.09 (s, 6H). m/z = 238.2 [M+H]+, 279.2 [M+H+ACN]+.
Figure imgf000155_0001
B. Preparation of (S)-2-(1 ,3-dioxoisoindolin-2-yl)-3-hvdroxypropanoic acid 48
[0352] (S)-2-amino-3-hydroxypropanoic acid 46 (2.00 g, 19.0 mmol, 1.00 equiv) and sodium carbonate (2.00 g, 19.0 mmol, 1.00 equiv) are dissolved in water (20 mL, 0.95 M). /V-Ethoxycarbonylphthalimide 47 (4.18 g, 19.0 mmol, 1.00 equiv), previously ground to a fine powder, is added at rt and the resulting suspension is stirred at rt until no more solid can be seen. The pH is adjusted to 1 by adding a 1 N HCI, at 0 °C. Some precipitate then appeared in the solution. This aqueous layer is extracted with AcOEt. The organic extract is evaporated under reduced pressure and oil is obtained. The oil is taken up in the minimum amount of DCM (10 mL) and toluene (30 mL) is added. Crystallization occurred over 3 days and the resulting white solid is filtered off, washed with toluene and hexanes and dried under vacuum. The desired product 48 is obtained as white solid (1.9 g, 8.1 mmol, 42% yield).
[0353] 1H NMR (500 MHz, CDC ) d (ppm) 7.93-7.88 (m, 2H), 7.82-7.75 (m, 2H), 5.12 (dd, 1 H, J = 5.5 Hz, J = 3.5 Hz), 4.33 (dd, 1 H, J = 13.0 Hz, J = 5.5 Hz), 4.24 (dd, 1 H, J = 12.5 Hz, J = 3.5 Hz).
Figure imgf000155_0002
C. Preparation of (S)-/V-(4-(((fe/f-butyldimethylsilyl)oxy)methyl)phenyl)-2-(1 ,3- dioxoisoindolin-2-yl)-3-hvdroxypropanamide 49
[0354] Under anhydrous conditions, compound 48 (1.00 g, 4.25 mmol, 1.00 equiv), compound 45 (1.06 g, 4.46 mmol, 1.05 equiv) and HATU (1.70 g, 4.46 mmol, 1.05 equiv) are dissolved in anhydrous DMF (1 mL, 4 M). DIPEA (2.33 mL, 13.4 mmol, 3.00 equiv) is added and the reaction mixture is stirred at rt for 2 hours. The reaction mixture is then diluted in AcOEt. This organic layer is washed with 10% aqueous citric acid, with sat. aqueous NhUCI, with sat. aqueous NaHCCb and then with brine. Solvent is evaporated under reduced pressure and the residue is purified by normal phase chromatography. The desired product 49 is obtained (0.98 g, 2.2 mmol, 51% yield).
[0355] 1H NMR (500 MHz, CDCIs) d (ppm) 8.99 (s, 1 H), 7.93-7.87 (m, 2H), 7.80- 7.75 (m, 2H), 7.30-7.25 (m, 2H), 6.82-6.76 (m, 2H), 5.20-5.07 (m, 1 H), 4.68 (s, 2H), 4.65 (s, 2H).
Figure imgf000156_0001
D. Preparation of (S)-2-(1-(4-(((fe/f-butyldimethylsilyl)oxy)methyl)phenyl)-2- oxoazetidin-3-yl)isoindoline-1 ,3-dione 50
[0356] Under anhydrous conditions, compound 49 (0.98 g, 2.16 mmol, 1.00 equiv), and triphenylphosphine (0.62 g, 2.4 mmol, 1.1 equiv) are dissolved in anhydrous THF (30 mL, 0.072 M). DIAD (0.48 g, 2.4 mmol, 1.1 equiv) is added at rt and the resulting reaction solution is stirred overnight at rt. The reaction mixture is diluted in AcOEt. This organic layer is washed with 10% aqueous citric acid, sat. aqueous NH4CI, sat. aqueous NaHC03 and then with brine. The washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is purified by column chromatography on silica gel. The desired phenyl monobactam product 50 is obtained as white solid (1.04 g), which is contaminated with some DIAD by-product. This material is used without further purification in the next step.
[0357] 1H NMR (500 MHz, CDCIs) d (ppm) 7.92-7.87 (m, 2H), 7.80-7.76 (m, 2H), 7.43-7.38 (m, 2H), 7.37-7.32 (m, 2H), 5.60 (dd, 1 H, J = 6.0 Hz, J = 3.5 Hz), 4.74 (s, 2H), 4.10-4.02 (m, 2H), 0.95 (s, 9H), 0.11 (s, 6H).
Figure imgf000157_0001
E. Preparation of (S)-3-amino-1 -(4-(((fe/f-butyldimethylsilyl)oxy)methyl)- phenyl)azetidin-2-one 52
[0358] Phenyl monobactam substrate 50 (1 .04 g, 2.38 mmol, 1 .00 equiv) is suspended in anhydrous EtOH (10 mL, 0.24 M). Ethylenediamine 51 (0.476 mL, 7.15 mmol, 3.00 equiv) is added at rt. THF (5 mL, 0.5 M) is added to dissolve solids present in the suspension. The resulting solution is stirred overnight at rt. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NH4CI, sat. aqueous NaHC03 and then with brine. The organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The residue obtained is purified by column chromatography on silica gel. The desired monobactam product 52 is obtained as white solid (0.37 g, 1 .2 mmol, 51% yield).
[0359] 1H NMR (500 MHz, CDCIs) d (ppm) 7.38-7.28 (m, 4H), 4.71 (s, 2H), 4.35 (br s, 1 H), 3.95 (dd, 1 H, J = 6.0 Hz, J = 5.0 Hz), 3.38 (dd, 1 H, J = 6.0 Hz, J = 2.5 Hz), 1 .75 (br s, 2H), 0.94 (s, 9H), 0.09 (s, 6H).
Figure imgf000157_0002
F. Preparation of (S,ZHe/f-butyl 2-(((1 -(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)- 2-((1 -(4-(((fe/f-butyldimethylsilyl)oxy)methyl)phenyl)-2-oxoazetidin-3-yl)amino)-2- oxoethylidene)-amino)oxy)-2-methylpropanoate 53
[0360] Under a nitrogen atmosphere, carboxylic acid 8 (0.547 g, 1 .27 mmol, 1 .05 equiv) is dissolved in anhydrous DMA (6 ml_, 0.2 M). At 5 °C, methanesulfonyl chloride (0.102 ml_, 1 .30 mmol, 1 .05 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.180 ml_, 1.30 mmol, 1.05 equiv). The resulting pale yellow suspension is stirred at 5 °C for 1 hour.
[0361] In parallel, a solution of substrate 52 (0.370 g, 1 .21 mmol. 1 .00 equiv) and /V-methylmorpholine (0.133 ml_, 1 .21 mmol, 1 .00 equiv) is prepared in anhydrous THF (4.0 ml_, 0.30 M).
[0362] At 5 °C, the solution of 52 is added dropwise to the solution of mesylate. The resulting reaction mixture is stirred at 5 °C for 1 hour. The reaction mixture is then diluted in AcOEt (50 ml_). This organic layer is washed with 10% aqueous citric acid (20 ml_), sat. aqueous NaHCCb (40 ml_) and then with brine. The washed organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The desired product 53 is obtained as white solid (0.81 g, 1 .1 mmol, 89% yield). This material is used without further purification in the next step.
[0363] m/z = 718.4 [M+H]+.
Figure imgf000158_0001
G. Preparation of (S.Z)-2-(((1 -(2-aminothiazol-4-yl)-2-((1 -(4- (hvdroxymethyl)phenyl)-2-oxoazetidin-3-yl)amino)-2-oxoethylidene)amino)oxy)-2- methylpropanoic acid 54 (476)
[0364] At rt, a liquid mixture of triethylsilane (0.025 mL), water (0.03 mL) and TFA (1 .0 mL) is prepared. Substrate 53 (0.050 g, 0.070 mmol, 1 .0 equiv) is dissolved in the mixture, at rt. The resulting solution is stirred at rt for 0.5 hour. Solvent is evaporated under reduced pressure and the residue is triturated twice in a 1 : 2 v/v diethyl ether/hexanes solution. The solid residue is purified by reverse phase column chromatography. The desired alcohol 54 (476) is obtained (0.003 g, 0.007 mmol, 10% yield).
[0365] 1H NMR (500 MHz, Acetonitrile-ds) d (ppm) 7.85 (d, 1 H, J = 8.0 Hz), 7.40- 7.32 (m, 4H), 7.02 (s, 1 H), 5.17-5.10 (m, 1 H), 4.02 (dd, 1 H, J = 5.5 Hz, J = 6.0 Hz), 3.74 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1.61 (s, 6H). m/z = 448.3 [M+H]+.
Figure imgf000159_0001
H. Preparation of (S,Z)-2-(((1 -(2-aminothiazol-4-yl)-2-((1 -(4-(4- methoxybenzyl)phenyl)-2-oxoazetidin-3-yl)amino)-2-oxoethylidene)amino)oxy)-2- methylpropanoic acid 55 (477)
[0366] Anisole (0.10 mL, 0.7 M) and TFA (1.0 mL, 0.07 M) are mixed together. The resulting solution is used to dissolve compound 53 (0.050 g, 0.070 mmol, 1.00 equiv). The resulting solution is stirred at rt for 0.5 hour. Solvent is evaporated under reduced pressure and the residue is triturated twice in a 1 : 2 v/v diethyl ether/hexanes mixture. The desired product 55 (477) is obtained as white solid (0.014 g, 0.026 mmol, 37% yield).
[0367] 1H NMR (500 MHz, Acetonitrile-ds) d (ppm) 8.17 (br s, 1 H), 7.88 (d, 1 H, J = 8.5 Hz), 7.33-7.26 (m, 2H), 7.23-7.17 (m, 2H), 7.13-7.07 (m, 2H), 7.02 (s, 1 H), 6.86- 6.81 (m, 2H), 5.14-5.06 (m, 1 H), 3.99 (dd, 1 H, J = 6.0 Hz, J = 6.0 Hz), 3.87 (s, 2H), 3.74 (s, 3H), 3.70 (dd, 1 H, J = 6.0 Hz, J = 2.5 Hz), 1.61 (s, 6H). m/z = 538.3 [M+H]+.
Figure imgf000160_0001
I. Preparation of (S,ZHe/f-butyl 2-(((1 -(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)-
2-((1 -(4-(hvdroxymethyl)phenyl)-2-oxoazetidin-3-yl)amino)-2- oxoethylidene)amino)oxy)-2-methylpropanoate 56
[0368] Under nitrogen atmosphere, compound 53 (0.210 g, 0.293 mmol, 1 .00 equiv) is dissolved in anhydrous THF (4 ml_, 0.07 M), at rt. Tetra-n-butylammonium fluoride (1 .0 M solution in THF, 0.60 ml_, 0.60 mmol, 2.0 equiv) is added and the resulting solution is stirred at rt for 2 hours. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NaHCCb and then with brine. The washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The desired compound 56 is obtained (0.20 g) and this material is used without further purification in the next step.
[0369] 1H NMR (500 MHz, CDCIs) d (ppm) 8.28 (d, 1 H, J = 7.0 Hz), 7.36 (s, 5H), 5.33-5.28 (m, 1 H), 4.68 (s, 2H), 4.07 (dd, 1 H, J = 5.5 Hz, J = 6.0 Hz), 3.73 (dd, 1 H, J = 6.0 Hz, J = 2.5 Hz), 1 .61 (s, 3H), 1 .59 (s, 3H), 1 .54 (s, 9H), 1 .42 (s, 9H).
Figure imgf000160_0002
J. Preparation of (S Z)-tert-buM 2-(((1 -(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)-
2-((1 -(4-(chloromethyl)phenyl)-2-oxoazetidin-3-yl)amino)-2-oxoethylidene)amino)oxy)-
2-methylpropanoate 57
[0370] Under nitrogen atmosphere, compound 56 (0.20 g, 0.33 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (4 ml_, 0.08 M). Triethylamine (0.070 ml_, 0.50 mmol, 1 .5 equiv), methanesulfonyl chloride (0.057 g, 0.50 mmol, 1 .5 equiv) and DMAP (0.005 g, 0.04 mmol, 0.1 equiv) are added. The resulting reaction mixture is stirred at rt for 18 hours. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NaHCCb and then with brine. The washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is purified by column chromatography on silica gel, eluting with 0-25% AcOEt/DCM gradient. The desired compound 57 is obtained.
[0371] 1H NMR (500 MHz, CDCIs) d (ppm) 8.31 (br s, 1 H), 7.43-7.34 (m, 5H), 5.35- 5.28 (m, 1 H), 4.58 (s, 2H), 4.08 (dd, 1 H, J = 6.0 Hz, J = 5.5 Hz), 3.76 (dd, 1 H, J = 6.0 Hz, J = 2.5 Hz), 1 .62 (s, 3H), 1 .61 (s, 3H), 1 .54 (s, 9H), 1 .42 (s, 9H). m/z = 622 [M+H]+.
Figure imgf000161_0001
K. Preparation of (S,Z)-2-(4-(3-(2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)benzyl)-6.7-dihvdroxyisoquinolin-2-ium 58 (478)
[0372] Compound 57 (0.050 g, 0.080 mmol, 1 .00 equiv), 6,7-dihydroxyisoquinolin- 2-ium bromide 12 (0.025 g, 0.10 mmol, 1 .3 equiv), caesium carbonate (0.017 g, 0.053 mmol, 0.65 equiv) and potassium iodide (0.013 g, 0.080 mmol, 1 .0 equiv) are dissolved in anhydrous DMF (0.8 mL, 0.1 M). The resulting solution is stirred at rt for 2 hours and then at 50 °C for 3 hours. The reaction mixture is purified by C18 reverse phase column chromatography. Fractions containing the desired intermediate are combined and lyophilized. The desired intermediate is obtained (0.020 g, m/z = 747.5 [M]+). The latter is dissolved in a mixture of triethylsilane (0.025 mL), water (0.03 mL) and TFA (1.0 mL), at 0 °C. The resulting reaction mixture is stirred at 0 °C for 3.5 hours and then 0.5 hour at rt. Solvent is evaporated under reduced pressure and the residue is triturated twice in diethyl ether. The desired compound 58 (478) is obtained as off-white solid (0.012 g, 0.02 mmol, 75% yield over 2 steps). [0373] 1H NMR (500 MHz, Acetonitrile-ds + D2O + 1 % TFA) d (ppm) 9.13 (s, 1 H), 8.12-8.08 (m, 1 H), 7.96 (d, 1 H, J = 7.0 Hz), 7.58 (s, 1 H), 7.50-7.40 (m, 5H), 7.00 (s, 1 H), 5.62 (s, 2H), 5.07 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 4.02 (dd, 1 H, J = 6.0 Hz, J = 5.5 Hz), 3.79 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .57 (s, 6H). m/z = 591.5 [M]+, 296.3 [M+H]2+.
Figure imgf000162_0001
L. Preparation of (S,Z)-1 -(4-(3-(2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)benzyl)-6,7-dihvdroxyquinolin-1 -ium 60 (479)
[0374] Compound 57 (0.100 g, 0.161 mmol, 1 .00 equiv), 6,7-dihydroxyquinolin-1 - ium bromide 59 (0.050 g, 0.21 mmol, 1 .3 equiv), caesium carbonate (0.034 g, 0.1 1 mmol, 0.65 equiv) and potassium iodide (0.027 g, 0.21 mmol, 1 .0 equiv) are dissolved in anhydrous DMF (1 .6 mL, 0.10 M), at rt. The resulting solution is stirred at 50 ° C for 18 hours. The reaction solution is purified by C18 reverse phase chromatography. Fractions containing the desired intermediate are combined and lyophilized. The intermediate so obtained (0.020 g, m/z = 747.3 [M]+) is dissolved in TFA (1 .0 mL) and the resulting solution is stirred at rt for 0.5 hour. Solvent is evaporated under reduced pressure. The residue is taken up in DMSO (0.5 mL) and is purified by C18 reverse phase chromatography. Fractions containing the desired compound are combined and lyophilized. The desired product 60 (479) is obtained as a white solid (0.004 g, 0.007 mmol, 4% yield over 2 steps).
[0375] 1H NMR (500 MHz, Acetonitrile-ds + D2O +1 % TFA) d (ppm) 8.81 (dd, 1 H, J = 6.5 Hz, J = 1 .5 Hz), 8.72 (dd, 2H, J = 7.0 Hz, J = 1 .5 Hz), 8.58 (tt, 1 H, J = 7.5 Hz, J = 1 .5 Hz), 8.03 (br t, 1 H, J = 7.0 Hz), 7.70 (dd, 1 H, J = 8.5 Hz, J = 6.0 Hz), 7.52 (s, 1 H), 7.50 (s, 1 H), 7.40-7.35 (m, 2H), 7.32-7.27 (m, 2H), 7.05 (s, 1 H), 5.91 (s, 2H), 5.05 (dd, 1 H, J = 5.5 Hz, J = 2.5 Hz), 4.01 (dd, 1 H, J = 6.0 Hz, J = 5.5 Hz), 3.77 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .54 (s, 6H). m/z = 591 .3 [M]+, 296.2 [M+H]2+.
Figure imgf000163_0001
M. Preparation of (S,Z)-1 -(4-(3-(2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)benzyl)pyridin-1 -ium 61 (481 )
[0376] Under inert atmosphere, compound 57 (0.050 g, 0.080 mmol, 1 .0 equiv) is dissolved in DCM (1 ml_, 0.08 M) and pyridine (0.014 ml_, 0.18 mmol, 2.2 equiv) is added, followed by DMAP (0.001 g, 0.008 mmol, 0.1 equiv). The resulting solution is stirred overnight at rt. DCM is evaporated under reduced pressure and potassium iodide (0.013 g, 0.080 mmol, 1 .0 equiv) and DMF (1 ml_, 0.08 M) are added to the reaction mixture. The resulting solution is stirred at rt for 2 hours and then at 50 °C for 1 hour. The reaction mixture is purified by C18 reverse phase column chromatography. Fractions containing the desired intermediate are combined and lyophilized. The desired intermediate is obtained (0.007 g, m/z = 665.4 [M]+). The intermediate is dissolved in TFA (1.0 ml_, 0.01 M) at rt and the resulting solution is stirred at rt for 1 .5 hours. Solvent is evaporated under reduced pressure and the residue is triturated twice in diethyl ether. The solid obtained is purified by C18 reverse phase column chromatography. Fractions containing the desired product are combined and lyophilized. The desired product 61 (481 ) is obtained as white solid (0.005 g, 0.01 mmol, 12% yield over 2 steps).
[0377] 1H NMR (500 MHz, Acetonitrile-ds + D2O + 1 % TFA) d (ppm) 8.77 (dd, 2H, J = 6.5 Hz, J = 1 .0 Hz), 8.53-8.47 (m, 1 H), 8.01 (dd, 2H, J = 7.0 Hz, J = 7.0 Hz), 7.50- 7.42 (m, 4H), 6.99 (s, 1 H), 5.68 (s, 2H), 5.07 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 4.03 (dd, 1 H, J = 6.0 Hz, J = 6.0 Hz), 3.81 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .57 (s, 3H), 1.56 (s, 3H). m/z = 509.1 [M]+. Example 5: Preparation of Exemplary Monobactams of Formula I: Compounds 489, 490 491 and 495
Figure imgf000164_0001
A. Preparation of (S)-2-(1 ,3-dioxoisoindolin-2-yl)-3-hvdroxypropanoic acid 48
[0378] Compound 48 is prepared from (S)-2-amino-3-hydroxypropanoic acid 46 following the procedure described in Example 4, Procedure B.
[0379] 1H NMR (500 MHz, CDC ) d (ppm) 7.93-7.88 (m, 2H), 7.82-7.75 (m, 2H), 5.12 (dd, 1 H, J = 5.5 Hz, J = 3.5 Hz), 4.33 (dd, 1 H, J = 13.0 Hz, J = 5.5 Hz), 4.24 (dd, 1 H, J = 12.5 Hz, J = 3.5 Hz).
Figure imgf000164_0002
B. Preparation of tert- butyl 2-amino-2-(diethoxyphosphoryl)acetate 64
[0380] Under inert atmosphere, sodium hydride (60% dispersion in mineral oil) (0.436 g, 10.9 mmol, 1.10 equiv) is suspended in anhydrous THF (20 mL, 0.55 M). This suspension is cooled to 0 °C. At this temperature, tert- butyl diethylphosphonoacetate 62 (from AK Scientific) (2.50 g, 9.90 mmol, 1 .00 equiv) is added and the stirring is continued for 0.5 hour at 0 °C. The resulting solution is transferred via cannula to a stirred suspension of 0-(diphenylphosphinyl)hydroxylamine 63 (2.31 g, 9.90 mmol, 1 .00 equiv) in anhydrous THF (20 mL, 0.50 M), at - 78 °C. The resulting mixture is stirred for 1 hour at - 78 °C and then warmed to rt and stirred at rt for 2 hours. The reaction mixture is diluted in AcOEt (200 mL). This organic layer is washed with 10% aqueous citric acid (3 x 80 mL). The combined aqueous washes are brought to pH 8-9 by addition of sodium carbonate, and are then extracted using DCM. The DCM extract is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The desired compound 64 is obtained as colourless oil (1 .84 g, 6.89 mmol, 70% yield).
[0381] 1H NMR (300 MHz, CDCb) d (ppm) 4.25-4.10 (m, 4H), 3.85 (d, 1 H, J = 20.1 Hz), 1 .52 (s, 9H), 1 .42-1.32 (m, 6H). m/z = 268.4 [M+H]+.
Figure imgf000165_0001
C. Preparation of tert- butyl 2-(diethoxyphosphoryl)-2-((S)-2-(1 ,3-dioxoisoindolin-2- yl)-3-hvdroxypropanam ido)acetate 65
[0382] Under anhydrous conditions, compound 48 (1 .43 g, 6.07 mmol, 1 .00 equiv), compound 64 (1 .70 g, 6.37 mmol, 1 .05 equiv) and DIPEA (3.13 mL, 18.0 mmol, 3.00 equiv) are dissolved in anhydrous DMF (12 mL, 0.51 M). HATU (2.42 g, 6.37 mmol, 1 .05 equiv) is added and the resulting mixture is stirred at rt for 4 hours. The reaction mixture is diluted in AcOEt. This organic layer is washed with 10% aqueous citric acid, with sat. aqueous NH4CI, with sat. aqueous NaHCCb and then with brine. The organic layer is dried over sodium sulfate. Solvent is evaporated from the dried organic layer under reduced pressure and the residue is purified by normal phase chromatography. The desired product 65 is obtained as white solid foam (as a mixture of 2 diastereoisomers, 2.43 g, 5.02 mmol, 83% yield).
[0383] 1H NMR (500 MHz, CDCb) d (ppm) 7.90-7.84 (m, 2H), 7.79-7.73 (m, 2H), 7.54 and 7.44 (2 d, 1 H), 5.10 and 5.06 (2 d, 1 H, J = 9.0 Hz), 5.04-4.98 (m, 1 H), 4.55- 4.49 (m, 1 H), 4.24-4.10 (m, 4H), 4.10-4.03 (m, 1 H), 3.57 and 3.45 (2 t, 1 H), 1 .50 and 1 .48 (2 s, 9H), 1 .37-1 .29 (m, 6H).
Figure imgf000166_0001
D. Preparation of tert- butyl 2-(diethoxyohosohoryl)-2-((S)-3-(1 ,3-dioxoisoindolin-2- yl)-2-oxoazetidin-1 -vDacetate 66
[0384] Under anhydrous conditions, compound 65 (2.43 g, 5.02 mmol, 1 .00 equiv) is dissolved in anhydrous THF (30 mL, 0.17 M). Triphenylphosphine (1 .97 g, 7.53 mmol, 1 .50 equiv) is added, followed by di-fe/f-butyl azodicarboxylate (1 .73 g, 7.53 mmol, 1 .50 equiv). The resulting mixture is stirred at rt for 4 hours. Solvent is evaporated under reduced pressure. The residue is purified by normal phase column chromatography. The product compound as purified still had some triphenylphosphine oxide residues. The product compound is dissolved in diethyl ether (10 mL) and left at - 20 °C for one night. Some triphenylphosphine oxide crystallized out of the solution and is removed by filtration. The filtrate still contained triphenylphosphine oxide. The filtrate is adsorbed onto silica gel prior to normal phase chromatography eluting with a AcOEt/hexanes gradient. The desired compound 66 is obtained (mixture of 2 diastereoisomers, 0.780 g, 1 .67 mmol, 33% yield).
[0385] 1H NMR (500 MHz, CDC ) d (ppm) 7.90-7.82 (m, 2H), 7.77-7.72 (m, 2H), 5.15-5.09 and 5.49-5.42 (2 m, 1 H), 5.15 and 5.05 (2 d, 1 H, J = 10.0 Hz), 4.36-4.25 (m,
1 H), 4.27-4.15 (m, 4H), 4.08-4.02 and 4.01 -3.96 (2 m, 1 H), 1 .56 and 1 .53 (2 s, 9H).
Figure imgf000166_0002
E. Preparation of 4-(((fe/f-butyldimethylsilyl)oxy)methyl)benzaldehyde 69
[0386] Compound 68 is prepared by a literature procedure (Wang et al. (2017) Org. Lett. 19(3):632-635). More specifically, under inert atmosphere, terephthalaldehyde 67 (10.0 g, 74.6 mmol, 1 .00 equiv) is dissolved in anhydrous THF (124 mL, 0.60 M) and ethanol (41 mL, 1.8 M). At 0 °C, sodium borohydride (0.705 g, 18.6 mmol, 0.25 equiv) is added in small portions, over 20 minutes. The resulting mixture is stirred at 0 °C for 2.5 hours. More sodium borohydride (0.039 g, 1 .0 mmol, 0.014 equiv) is added at 0 °C. The resulting mixture is stirred at 0 °C for 2 hours. The reaction is quenched by addition of aqueous 2M HCI (10 mL). The resulting yellow solution, containing white solids is kept at 0 °C for the night. Solvent is evaporated under reduced pressure. Water (150 mL) is then added to the residue and the product is extracted with AcOEt (2 x 100 mL). Organic extracts are combined and washed with brine. The organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is purified by column chromatography on silica gel (Biotage SNAP, 100 g) eluting with 0-50% AcOEt/hexanes gradient. The desired product 68 is obtained as a white solid (6.68 g, 49.1 mmol, 66% yield).
[0387] 1H NMR (500 MHz, CDCIs) d (ppm) 10.02 (s, 1 H), 7.92-7.85 (m, 2H), 7.57- 7.52 (m, 2H), 4.82 (d, 2H, J = 5.5 Hz), 1 .85 (t, 1 H, J = 6.0 Hz).
[0388] Under inert atmosphere, compound 68 (1 .00 g, 7.35 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (40 mL, 0.18 M). TBDMSCI (1 .33 g, 8.82 mmol, 1 .20 equiv) is added, followed by imidazole (0.600 g, 8.82 mmol, 1 .20 equiv) and the resulting mixture is stirred at rt for 1 hour. The reaction mixture is diluted in AcOEt. This organic layer is washed with 10% aqueous citric acid, sat. aqueous NH4CI and then with brine. The washed organic layer is dried over sodium sulfate, filtered and solvent is evaporated under reduced pressure. The desired product 69 is obtained as colourless oil (1 .8 g, 7.2 mmol, 98% yield).
[0389] 1H NMR (500 MHz, CDCIs) d (ppm) 10.0 (s, 1 H), 7.87-7-82 (m, 2H), 7.52- 7.47 (m, 2H), 4.82 (s, 2H), 0.96 (s, 9H), 0.12 (s, 6H).
Figure imgf000168_0001
F. Preparation of (S,EHe/f-butyl 3-(4-(((fe/f-butyldimethylsilyl)oxy)methyl)phenyl)-
2-(3-(1 ,3-dioxoisoindolin-2-yl)-2-oxoazetidin-1 -yl)acrylate 70
[0390] Under a nitrogen atmosphere, aldehyde 69 (0.438 g, 1.75 mmol, 1.20 equiv) and phosphonate 66 (0.680 g, 1.46 mmol, 1.00 equiv) are dissolved in anhydrous THF (10 ml_ 0.15 M). At 0 °C, sodium hydride (60% suspension in mineral oil) (0.064 g, 1.6 mmol, 1.1 equiv) is added. The resulting mixture is stirred at 0 °C for 0.5 hour and then at rt for 2 hours. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NFUCI and then with brine. The washed organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The yellow oil obtained is purified by column chromatography on silica gel. The desired compound 70 (pure trans double bond) is obtained as white solid (0.300 g, 0.533 mmol, 37% yield).
[0391] 1H NMR (500 MHz, CDCIs) d (ppm) 7.90-7.85 (m, 2H), 7.80-7.74 (m, 2H), 7.30-7.24 (m, 4H), 7.03 (s, 1 H), 5.54 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 4.73 (s, 2H), 4.08 (dd, 1 H, J = 6.5 Hz, J = 6.0 Hz), 4.02 (dd, 1 H, J = 6.0 Hz, J = 3.5 Hz), 1.41 (s, 9H), 0.95 (s, 9H), 0.11 (s, 6H).
Figure imgf000168_0002
G. Preparation of (S,E)-tert- butyl 2-(3-amino-2-oxoazetidin-1 -yl)-3-(4-(((fe/f- butyldimethylsilyl)oxy)methyl)phenyl)acrylate 71
[0392] Under inert atmosphere, compound 71 (0.300 g, 0.534 mmol, 1 .00 equiv) is dissolved in anhydrous ethanol (2.0 ml_, 0.27 M). At rt, ethylenediamine 51 (0.1 1 ml_, 1 .6 mmol, 3.0 equiv) is added. The resulting solution is stirred at rt for 2 hours. The reaction solution is adsorbed onto silica gel, dried under vacuum and then purified by column chromatography on silica gel eluting with a 0-10% MeOH/DCM gradient. The desired free amine 71 is obtained as white solid (0.20 g, 0.46 mmol, 87% yield).
[0393] 1H NMR (500 MHz, CDCIs) d (ppm) 7.25-7.20 (m, 4H), 6.69 (s, 1 H), 4.72 (s, 2H), 4.32-4.25 (m, 1 H), 3.89 (dd, 1 H, J = 6.5 Hz, J = 5.5 Hz), 3.36 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 1 .42 (s, 9H), 0.93 (s, 9H), 0.09 (s, 6H).
Figure imgf000169_0001
H. Preparation of (E)-tert- butyl 2-((S)-3-((Z)-2-(((1 -(fe/f-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2- oxoazetidin-1 -yl)-3-(4-(((fe/f-butyldimethylsilyl)oxy)methyl)phenyl)acrylate 72
[0394] Under inert atmosphere, carboxylic acid 8 (0.209 g, 0.487 mmol, 1 .05 equiv) is dissolved in anhydrous DMA (3.0 ml_, 0.16 M). At 5 °C, methanesulfonyl chloride (0.040 ml_, 0.51 mmol, 1 .1 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.071 ml_, 0.51 mmol, 1 .1 equiv). The resulting pale yellow suspension is stirred at 5 °C for 1 hour.
[0395] In parallel, a solution of substrate 71 (0.200 g, 0.463 mmol. 1 .00 equiv) and /V-methylmorpholine (0.051 ml_, 0.46 mmol, 1 .00 equiv) is prepared in anhydrous THF (3.0 ml_, 0.15 M).
[0396] At 5 °C, the solution of 71 is added dropwise to the solution of mesylate. The resulting solution is stirred at 5 °C for 1 hour. The reaction mixture is diluted in AcOEt (50 ml_). This organic layer is washed with 10% aqueous citric acid (20 ml_), sat. aqueous NaHCCb (40 mL) and then with brine. The washed organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The desired product 72 is obtained as a white solid (0.37 g, 0.44 mmol, 95% yield). This material is used without further purification in the next step.
[0397] m/z = 844.0 [M+H]+.
Figure imgf000170_0001
I. Preparation of (E)-tert- butyl 2-((S)-3-((Z)-2-(((1 -(fe/f-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2- oxoazetidin-1 -yl)-3-(4-(hvdroxymethyl)phenyl)acrylate 73
[0398] Under nitrogen atmosphere, compound 72 (0.390 g, 0.460 mmol, 1 .00 equiv) is dissolved in anhydrous THF (5 mL, 0.09 M), at rt. At 0 °C, tetra-n- butylammonium fluoride (1 .0 M solution in THF, 0.69 mL, 0.69 mmol, 1 .5 equiv) is added and the resulting solution is stirred at 0 °C for 1 hour and then overnight at rt. Solvent is evaporated under reduced pressure until ca. 2 mL of THF remains. This solution is adsorbed on silica gel on top of a chromatography column, and is purified. The desired compound 73 is obtained (0.17 g, 0.23 mmol, 51% yield).
[0399] 1H NMR (500 MHz, CDC ) d (ppm) 8.16 (m, 1 H), 7.32-7.25 (m, 4H), 5.29- 5.24 (m, 1 H), 4.70 (d, 1 H, J = 6.0 Hz), 4.04 (dd, 1 H, J = 6.0 Hz, J = 5.5 Hz), 3.73 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .60 (s, 6H), 1 .54 (s, 9H), 1 .46 (s, 9H), 1 .43 (s, 9H). m/z = 730.2 [M+H]+.
Figure imgf000171_0001
73 74
J. Preparation of (E)-tert- butyl 2-((S)-3-((Z)-2-(((1 -(fe/f-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2- oxoazetidin-1 -yl)-3-(4-(chloromethyl)phenyl)acrylate 74
[0400] Under nitrogen atmosphere, compound 73 (0.170 g, 0.233 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (4 ml_, 0.06 M). At 5 °C, methanesulfonyl chloride (0.027 ml_, 0.35 mmol, 1 .5 equiv) is added dropwise, followed by dropwise addition of /V-methylmorpholine (0.033 ml_, 0.30 mmol, 1 .3 equiv). The resulting pale yellow solution is stirred at 0 °C for 2 hours. DMAP (0.002 g) is added and the resulting solution is stirred at rt for 1 hour. Solvent is evaporated under reduced pressure until ca. 1 ml_ remains. Anhydrous DMF (3 ml_, 0.08 M) is added to the residue, followed by lithium chloride (0.020 g, 0.47 mmol, 2.0 equiv). The resulting suspension is stirred at rt for 2 hours. The reaction mixture is diluted in AcOEt. This organic layer is washed with sat. aqueous NhUCI and then with brine. The washed organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is purified by normal phase chromatography. The desired compound 74 is obtained as white solid (0.1 1 g, 0.15 mmol, 63% yield).
[0401] 1H NMR (500 MHz, CDCIs) d (ppm) 8.29 (brs, 1 H), 8.19 (d, 1 H, J = 8.0 Hz), 7.35 (s, 1 H), 7.34-7.31 (m, 2H), 7.27-7.23 (m, 2H), 6.89 (s, 1 H), 5.28-5.22 (m, 1 H), 4.58 (s, 2H), 4.04 (dd, 1 H, J = 6.5 Hz, J = 5.5 Hz), 3.74 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .61 (2 s, 6H), 1.53 (s, 9H), 1 .45 (s, 9H), 1.41 (s, 9H). m/z = 748.5 [M+H]+.
Figure imgf000172_0001
K. Preparation of (E)-2-((S)-3-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)-3-(4-(chloromethyl)phenyl)acrylic acid 75 (489)
[0402] Under inert atmosphere, compound 74 (0.010 g, 0.013 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (0.5 mL, 0.03 M). At rt, TFA (0.5 mL, 0.03 M) is added and the resulting solution is stirred at rt for 1 .5 hours. Solvents are evaporated under reduced pressure and the residue is triturated twice in diethyl ether. The desired product 75 (489) is obtained as white solid (0.007 g, 0.013 mmol, quantitative yield).
[0403] 1H NMR (500 MHz, Acetonitrile-ds) d (ppm) 7.90 (d, 1 H, J = 8.0 Hz), 7.43- 7.36 (m, 2H), 7.37-7.29 (m, 2H), 7.01 (s, 1 H), 6.58 (s, 1 H), 5.18-5.12 (m, 1 H), 4.68 (s, 2H), 3.98 (dd, 1 H, J = 6.0 Hz, J = 5.5 Hz), 3.78 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 1 .61 (s, 3H), 1 .60 (s, 3H). m/z = 535.9 [M+H]+.
Figure imgf000172_0002
L. Preparation of 2-(4-((a-2-((S)-3-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)-2-carboxyvinyl)benzyl)- 6.7-dihvdroxyisoguinolin-2-ium 76 (490)
[0404] Compound 74 (0.020 g, 0.027 mmol, 1 .00 equiv), 6,7-dihydroxyisoquinolin- 2-ium bromide 12 (0.008 g, 0.035 mmol, 1 .3 equiv), caesium carbonate (0.006 g, 0.018 mmol, 0.65 equiv) and potassium iodide (0.004 g, 0.027 mmol, 1 .0 equiv) are dissolved in anhydrous DMF (1 mL, 0.03 M). The resulting solution is stirred at 50 °C for 18 hours. The reaction mixture is purified by C18 reverse phase column chromatography. Fractions containing the desired intermediate are combined and lyophilized. The desired intermediate is obtained (0.020 g, m/z = 873.6 [M]+). The latter is dissolved in anhydrous DCM (0.5 mL, 0.05 M). TFA (0.5 mL, 0.05 M) is added and the resulting solution is stirred at rt for 1 .5 hours. Solvents are evaporated under reduced pressure and the residue is triturated twice in diethyl ether. The desired compound 76 (GL340) is obtained as white solid (0.013 g, 0.020 mmol, 73% yield over 2 steps).
[0405] 1H NMR (500 MHz, Acetonitrile-ds + D2O + 1 % TFA) d (ppm) 9.19 (s, 1 H), 8.13 (d, 1 H, J = 7.0 Hz), 7.99 (d, 1 H, J = 7.0 Hz), 7.59 (s, 1 H), 7.46 (s, 1 H), 7.43-7.37 (m, 4H), 7.08 (s, 1 H), 6.55 (s, 1 H), 5.07 (dd, 1 H, J = 5.5 Hz, J = 2.5 Hz), 3.97 (dd, 1 H, J = 6.5 Hz, J = 6.0 Hz), 3.79 (dd, 1 H, J = 6.0 Hz, J = 2.5 Hz), 1 .59 (s, 3H), 1 .58 (s, 3H). m/z = 661 .3 [M]+, 331 .3 [M+H]2+.
Figure imgf000173_0001
M. Preparation of 1 -(4-((a-2-((S)-3-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)-2-carboxyvinyl)benzyl)-
5,6-dihvdroxy-2-methyl-2/-/-indazol-1 -ium 77 (491 )
[0406] Compound 74 (0.040 g, 0.054 mmol, 1 .00 equiv), 5,6-dihydroxy-2-methyl- 2/-/-indazol-1 -ium bromide 14 (0.017 g, 0.070 mmol, 1 .3 equiv), caesium carbonate (0.01 1 g, 0.035 mmol, 0.65 equiv) and potassium iodide (0.009 g, 0.054 mmol, 1 .0 equiv) are dissolved in anhydrous DMF (0.5 ml_, 0.1 M), at rt. The resulting solution is stirred at 50 ° C for 18 hours and then at 70 ° C for another 18 hours. The reaction solution is purified by C18 reverse phase chromatography. Fractions containing the desired intermediate are combined and lyophilized. The intermediate so obtained (0.020 g, m/z = 876.5 [M]+) is dissolved in DCM (0.5 ml_, 0.05 M). TFA (0.5 ml_, 0.05 M) is added and the resulting solution is stirred at rt for 1 .5 hours. Solvent is evaporated under reduced pressure. The residue is triturated twice in diethyl ether. The desired product 77 (491 ) is obtained as white solid (0.005 g, 0.008 mmol, 14% yield over 2 steps). [0407] 1H NMR (500 MHz, Acetonitrile-ds + D2O + 1 % TFA) d (ppm) 8.44 (s, 1 H), 7.38-7.33 (m, 2H), 7.09 (s, 1 H), 7.12-7.7.07 (m, 2H), 7.06-7.02 (m, 2H), 6.53 (s, 1 H), 5.73 (s, 2H), 5.07 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 4.08 (s, 3H), 3.95 (dd, 1 H, J = 6.5 Hz, J = 6.0 Hz), 3.78 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 1 .69 (s, 6H). m/z = 664.4 [M]+, 332.7 [M+H]2+.
Figure imgf000174_0001
N. Preparation of 2-(4-((a-2-((S)-3-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)-2-carboxyvinyl)benzyl)- 5, 6-dihvdroxy-1 -methyl-1 /-/-indazol-2-ium 79 (495)
[0408] Compound 74 (0.048 g, 0.064 mmol, 1 .00 equiv), 5,6-dihydroxy-1 -m ethyl - 1 /-/-indazol-2-ium bromide 78 (0.020 g, 0.084 mmol, 1 .3 equiv), caesium carbonate (0.014 g, 0.042 mmol, 0.65 equiv) and potassium iodide (0.01 1 g, 0.064 mmol, 1 .0 equiv) are dissolved in anhydrous DMF (0.64 ml_, 0.1 M), at rt. The resulting solution is stirred at 70 ° C for 66 hours. The reaction mixture is purified by C18 reverse phase chromatography. Fractions containing the desired intermediate are combined and lyophilized. The intermediate so obtained (0.020 g, m/z = 876.6 [M]+) is dissolved in DCM (0.5 ml_, 0.05 M). TFA (0.5 ml_, 0.05 M) is added and the resulting solution is stirred at rt for 1 .5 hours. Solvent is evaporated under reduced pressure. The residue is triturated twice in diethyl ether. The desired product 79 (495) is obtained as white solid (0.007 g, 0.01 1 mmol, 16% yield over 2 steps).
[0409] 1H NMR (500 MHz, Acetonitrile-ds + D2O + 1 % TFA) d (ppm) 8.41 (s, 1 H), 7.45-7.40 (m, 2H), 7.28-7.23 (m, 2H), 7.20 (s, 1 H), 7.05 (s, 1 H), 6.98 (s, 1 H), 6.56 (s,
1 H), 5.72 (s, 2H), 5.08 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 3.95 (dd, 1 H, J = 6.5 Hz, J = 6.0 Hz), 3.78 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 1 .57 (s, 6H). m/z = 664.4 [M]+, 332.8 [M+H]2+. Example 6: Synthesis of Exemplary Monobactams of Formula I: Compound 500
Figure imgf000175_0001
A. Preparation of (S)-2-(1 ,3-dioxoisoindolin-2-yl)-3-hvdroxypropanoic acid 48
[0410] Compound 48 is prepared from (S)-2-amino-3-hydroxypropanoic acid 46 following the procedure described in Example 4, Procedure B.
[0411] 1H NMR (500 MHz, CDC ) d (ppm) 7.93-7.88 (m, 2H), 7.82-7.75 (m, 2H), 5.12 (dd, 1 H, J = 5.5 Hz, J = 3.5 Hz), 4.33 (dd, 1 H, J = 13.0 Hz, J = 5.5 Hz), 4.24 (dd, 1 H, J = 12.5 Hz, J = 3.5 Hz).
G-L © Q
G N-NH3 CI
Figure imgf000175_0002
B. Preparation of (S)-2-(1 ,3-dioxoisoindolin-2-yl)-3-hvdroxy-/V-(pyrrolidin-1 - vDorooanamide 81
[0412] Under inert atmosphere, compound 48 (0.300 g, 1.28 mmol, 1.00 equiv) and pyrrolidin-1 -amonium chloride 80 (from Enamine) (0.164 g, 1.34 mmol, 1.05 equiv) are dissolved in anhydrous DMF (4.3 mL, 0.30 M). Diisopropylethylamine (0.455 mL, 2.62 mmol, 2.05 equiv) is then added, followed by HATU (0.485 g, 1.28 equiv, 1.00 equiv). The resulting yellow solution is stirred at rt for 3.5 hours. The reaction mixture is diluted in AcOEt (25 mL) and this organic solution is washed with water (20 mL) and sat. aqueous NaHCCb (20 mL). Solid sodium bicarbonate is added to the aqueous wash until saturated. This aqueous layer is extracted with AcOEt (20 mL). The organic extracts are combined and dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The desired product 81 is obtained with other small contaminants (e.g., residual DMF, tetramethyl urea) as a yellow oil (0.350 g, m/z = 304.2 [M+H]+). This material is used without further purification in the next step.
Figure imgf000176_0001
C. Preparation of (S)-2-(2-oxo-1 -(pyrrolidin-1 -yl)azetidin-3-yl)isoindoline-1 ,3-dione 82
[0413] Under inert conditions, compound 81 (0.350 g, theoretically 0.387 g, 1.28 mmol, 1.00 equiv) is dissolved in anhydrous DCM (12.8 mL, 0.100 M). At 0 °C, triphenylphosphine (0.351 g, 1.34 mmol, 1.05 equiv) is added in one portion, followed by addition in one portion of DTBAD (0.308 g, 1.34 mmol, 1.05 equiv). The resulting white suspension is stirred at rt for 15 minutes and then at rt for 24 hours. Solvent is evaporated under reduced pressure and the residue is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with a 0-40% AcOEt/DCM gradient. The fractions containing the desired compound are combined and solvent is evaporated under reduced pressure. The syrup obtained is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with 0-40% AcOEt/DCM. The desired product 82 is obtained as a colourless oil (0.124 g, 0.435 mmol, 34% yield over 2 steps).
[0414] 1H NMR (500 MHz, CDC ) d (ppm) 7.89-7.84 (m, 2H), 7.78-7.73 (m, 2H), 5.25 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 3.81 (dd, 1 H, J = 4.5 Hz, J = 2.5 Hz), 3.74 (dd, 1 H, J = 5.5 Hz, J = 5.0 Hz), 3.18-3.06 (m, 4H), 1.94-1.83 (m, 4H). m/z = 286.2 [M+H]+.
Figure imgf000176_0002
D. Preparation of (S)-2-(2-oxoazetidin-3-yl)isoindoline-1 ,3-dione 83
[0415] Under inert conditions, compound 82 (0.124 g, 0.435 mmol, 1 .00 equiv) is dissolved in MeOH (3.6 mL, 0.12 M). At rt, mefa-chloroperoxybenzoic acid (77% purity, 0.195 g, 0.869 mmol, 2.00 equiv) is added in one portion. The resulting colourless solution is stirred at rt for 4.5 hours. Some white solid precipitated out of the solution. The reaction mixture is evaporated to dryness. The white solid obtained is dissolved in AcOEt (20 mL) and this organic layer is washed with sat. aqueous NaHCCb. The washed organic layer is then dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The white solid obtained is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with 0-10% MeOH/DCM. The desired product 83 is obtained as a white solid (0.069 g, 0.32 mmol, 73% yield).
[0416] 1H NMR (300 MHz, CDCIs) d (ppm) 7.94-7.84 (m, 2H), 7.80-7.73 (m, 2H), 5.96 ( br s, 1 H), 5.52 (dd, 1 H, J = 5.7 Hz, J = 3.0 Hz), 3.80 (dd, 1 H, J = 5.4 Hz, J = 3.0 Hz), 3.73 (dd, 1 H, J = 5.4 Hz, J = 5.4 Hz).
Figure imgf000177_0001
E. Preparation of (Z)-fe/f-butyl 2-bromo-3-(4-(hvdroxymethyl)phenyl)acrylate 85
[0417] Under inert atmosphere, tert- butyl 2-(triphenylphosphoranylidene)acetate 84 (from Oakwood Chemicals) (0.508 g, 1.35 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (9.0 ml_, 0.15 M). At O °C, bromide (0.076 ml_, 1 .5 mmol, 1.1 equiv) is added dropwise. The resulting yellow solution is stirred at 0 °C for 45 minutes. The reaction mixture is diluted in DCM (15 ml_) and this organic layer is washed with water (15 ml_) and then sat. aqueous NaHCCb (20 ml_). The organic layer is dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The pale yellow syrup obtained is then dissolved in DCM (9.0 ml_, 0.15 M) and aldehyde 68 (preparation described in Example 5, Procedure E) (0.220, 1 .62 mmol, 1 .20 equiv) is added. The resulting solution is stirred at rt for 2 hours and 15 minutes. The reaction mixture is evaporated to dryness. The residue is purified by column chromatography on silica gel (Biotage SNAP, 25 g) eluting with 10-40% AcOEt/hexanes gradient. The desired product 85 is obtained as colourless oil (contaminated with ca. 12% of non- brominated alkene) (0.189 g, 0.603 mmol, 45% yield). This material is used without further purification in the next step.
[0418] 1H NMR (500 MHz, CDCIs) d (ppm) 8.12 (s, 1 H), 7.87-7.82 (m, 2H), 7.46- 7.40 (m, 2H), 4.75 (d, 2H, J = 6.0 Hz), 1 .72 (t, 1 H, J = 6.0 Hz), 1 .58 (s, 9H).
Figure imgf000178_0001
F. Preparation of (S,ZHe/f-butyl 2-(3-(1 ,3-dioxoisoindolin-2-yl)-2-oxoazetidin-1 -yl)-
3-(4-(hvdroxymethyl)phenyl)acrylate 86
[0419] Under a nitrogen atmosphere, compound 83 (0.069 g, 0.32 mmol, 1.0 equiv), potassium carbonate (0.088 g, 0.64 mmol, 2.0 equiv) and copper (I) iodide (0.015 g, 0.080 mmol. 0.25 equiv) are introduced into a vial. To that mixture, a solution of bromide 85 (0.1 10 g, 0.351 mmol, 1 .10 equiv) in anhydrous toluene (1 .6 ml_, 0.20 M) is added, under nitrogen atmosphere. 1 ,2-dimethylethylenediamine (DMEDA) (0.017 ml_, 0.16 mmol. 0.50 equiv) is quickly added, after which a twist cap is placed on the vial. The resulting blue suspension is immersed in a preheated oil bath (1 10 °C) and is stirred at this temperature for 1 .5 hours. Copper iodide (0.006 g, 0.03 mmol, 0.1 equiv) and DMEDA (0.007 g, 0.06 mmol, 0.2 equiv) are added and the mixture is stirred at 1 10 °C for 1 .5 hours. The mixture is cooled to rt and purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with 0-40% AcOEt/DCM gradient. The desired product 86 is obtained as off-white solid (0.080 g, 0.18 mmol, 56% yield).
[0420] 1H NMR (500 MHz, CDCIs) d (ppm) 7.95-7.90 (m, 2H), 7.85-7.80 (m, 2H), 7.80-7.75 (m, 2H), 7.50 (s, 1 H), 7.50-7.45 (m, 2H), 5.61 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 4.75 (s, 2H), 4.04 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 3.91 (dd, 1 H, J = 6.0 Hz, J = 6.0 Hz), 1 .59 (s, 9H).
Figure imgf000179_0001
G. Preparation of (S Z)-tert-buM 2-(3-amino-2-oxoazetidin-1 -yl)-3-(4- (hydroxymethvDohenvDacrylate 87
[0421] Under inert atmosphere, compound 86 (0.080 g, 0.18 mmol, 1.0 equiv) is dissolved in anhydrous ethanol (1.2 ml_, 0.15 M). At rt, ethylenediamine 51 (0.036 ml_, 0.53 mmol, 3.0 equiv) is added dropwise. The resulting solution is stirred at rt for 1 hour and 40 minutes. The reaction mixture is evaporated to dryness and the residue is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with a 0- 10% MeOH/DCM gradient. A second column chromatography is performed eluting with a 0-10% MeOH/DCM gradient. The desired product 87 is obtained as a white film (0.023 g, 0.072 mmol, 40% yield).
[0422] 1H NMR (500 MHz, MeOD + CDCIs) d (ppm) 7.50-7.45 (m, 2H), 7.45 (s, 1 H), 7.42-7.36 (m, 2H), 4.63 (s, 2H), 4.26 (dd, 1 H, J = 5.5 Hz, J = 2.5 Hz), 3.77 (dd, 1 H, J = 6.0 Hz, J = 6.0 Hz), 3.40 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1.55 (s, 9H). m/z = 341.2 [M+Na]+.
Figure imgf000179_0002
H. Preparation of (ZHe/f-butyl 2-((S)-3-((Z)-2-(((1 -(fe f-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((fe f-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2- oxoazetidin-1 -yl)-3-(4-(hvdroxymethyl)phenyl)acrylate 88
[0423] Under inert atmosphere, carboxylic acid 8 (0.037 g, 0.087 mmol, 1.20 equiv) is dissolved in anhydrous DCM (0.9 ml_, 0.1 M). At 0 °C, methanesulfonyl chloride (0.008 ml_, 0.1 mmol, 1.5 equiv) is added dropwise, followed by dropwise addition of triethylamine (0.012 ml_, 0.087 mmol, 1.2 equiv). The resulting pale yellow suspension is stirred at 0 °C for 50 minutes.
[0424] In parallel, a solution of substrate 87 (0.023 g, 0.072 mmol, 1.0 equiv) in anhydrous DCM (0.7 ml_, 0.1 M) is prepared. At 0 °C, it is added to the solution of mesylate, followed by /V-methylmorpholine (0.008 ml_, 0.07 mmol, 1 equiv). The resulting cloudy colourless solution is stirred between 0 °C and rt for 3.5 hours. The reaction mixture is diluted in DCM (20 ml_) and this organic layer is washed with sat. aqueous NhUCI (15 ml_). The washed organic layer s dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The desired product 88 is obtained as a white solid (0.035 g, 0.048 mmol, 66% yield). This material is used without further purification in the next step.
[0425] 1H NMR (300 MHz, CDCIs) d (ppm) 8.13 (d, 1 H, J = 7.8 Hz), 7.57-7.52 (m, 2H), 7.46 (s, 1 H), 7.43-7.38 (m, 2H), 7.35 (s, 1 H), 5.33-5.25 (m, 1 H), 4.72 (s, 2H), 3.91 (dd, 1 H, J = 5.7 Hz, J = 5.7 Hz), 3.78 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1.59 (s, 15H),
I .55 (s, 9H), 1.45 (s, 9H). m/z = 730.4 [M+H]+.
Figure imgf000180_0001
I. Preparation of (ZHe f-butyl 2-((S)-3-((Z)-2-(((1 -(fe/f-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((fe/f-butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2- oxoazetidin-1 -yl)-3-(4-(chloromethyl)phenyl)acrylate 88
[0426] Under nitrogen atmosphere, compound 88 (0.035 g, 0.048 mmol, 1.0 equiv) is dissolved in anhydrous DCM (1.5 ml_, 0.033 M), at rt. The mixture is cooled to 0 °C. Methanesulfonyl chloride (0.0060 ml_, 0.070 mmol, 1 .5 equiv) is added dropwise, followed by addition /V-methylmorpholine (0.0070 ml_, 0.062 mmol, 1 .3 equiv) and the resulting colourless solution is stirred at rt for 2 hours. Methanesulfonyl chloride (0.004 ml_, 0.05 mmol, 1 equiv) is added and the reaction mixture is stirred at rt for 3 hours. Solvent is evaporated under reduced pressure. Lithium chloride (0.0060 g, 0.14 mmol, 3.0 equiv) is added to the residue, followed by anhydrous DMF (0.5 mL, 0.1 M). The resulting pale yellow suspension is stirred at rt for 17 hours. The reaction mixture is diluted in AcOEt (20 mL) and this organic layer is washed with sat. aqueous NhUCI (2 x 10 mL). The washed organic layer is then dried over magnesium sulfate, filtered and solvent is evaporated under reduced pressure. The residue is purified by column chromatography on silica gel (Biotage SNAP, 10 g) eluting with a 10-40% AcOEt/hexanes gradient. The desired product 89 is obtained as a white solid (0.025 g, 0.033 mmol, 70% yield).
[0427] 1H NMR (300 MHz, CDCh) d (ppm) 8.15 (d, 1 H, J = 8.1 Hz), 7.60-7.54 (m, 2H), 7.44 (s, 1 H), 7.44-7.38 (m, 2H), 7.36 (s, 1 H), 5.35-5.25 (m, 1 H), 4.58 (s, 2H), 3.92 (dd, 1 H, J = 6.0 Hz, J = 5.7 Hz), 3.79 (dd, 1 H, J = 5.7 Hz, J = 3.0 Hz), 1 .61 (s, 3H), 1 .60 (s, 3H), 1 .58 (s, 9H), 1 .56 (s, 9H), 1 .46 (s, 9H). m/z = 748.6 [M+H]+.
Figure imgf000181_0001
J. Preparation of 2-(4-((Z)-3-(fe/f-butoxy)-2-((S)-3-((Z)-2-(((1 -(fe/f-butoxy)-2- methyl-1 -oxopropan-2-yl)oxy)imino)-2-(2-((fe/f-butoxycarbonyl)amino)thiazol-4- yl)acetamido)-2-oxoazetidin-1 -yl)-3-oxoprop-1 -en-1 -yl)benzyl)-6,7- dihvdroxyisoquinolin-2-ium 90
[0428] Under inert atmosphere, compound 89 (0.025 g, 0.033 mmol, 1.0 equiv), 6,7-dihydroxyisoquinolin-2-ium bromide 12 (0.01 1 g, 0.043 mmol, 1 .3 equiv) and potassium carbonate (0.0030 g, 0.022 mmol, 0.65 equiv) are suspended on anhydrous DMF (1 mL, 0.03 M). To the resulting suspension, potassium iodide (0.0060 g, 0.035 mmol, 1 .0 equiv) is added. The resulting orange suspension is immersed in a preheated oil bath (50 °C) and stirred at this temperature for 2 hours. Potassium iodide (0.003 g, 0.02 mmol, 0.5 equiv) is added and the suspension is stirred at 50 °C for 20 more minutes. The reaction mixture is cooled to rt and is purified by reverse phase column chromatography (Teledyne GOLD C18, 12 g) eluting with 5-100% ACN/H2O (+ 0.1 % TFA gradient). Fractions containing the desired product are combined and lyophilized. The desired product 90 is obtained as white solid (0.024 g, 0.027 mmol, 82% yield).
[0429] 1H NMR (500 MHz, Acetone-de) d (ppm) 9.55 (s, 1 H), 8.41 (d, 1 H, J = 7.0 Hz), 8.22 (d, 1 H, J = 8.0 Hz), 8.13 (d, 1 H, J = 7.0 Hz), 7.87-7.82 (m, 2H), 7.76 (s, 1 H), 7.61 (s, 1 H), 7.63-7.58 (m, 2H), 7.41 (s, 1 H), 7.36 (s, 1 H), 5.98 (s, 2H), 5.20-5.19 (m, 1 H), 3.98-3.93 (m, 2H), 1 .55 (s, 9H), 1 .54 (s, 9H), 1 .49 (s, 3H), 1 .47 (s, 3H), 1.42 (s, 9H). m/z = 873.7 [M]+.
Figure imgf000182_0001
K. Preparation of 2-(4-((Z)-2-((S)-3-((Z)-2-(2-aminothiazol-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-oxoazetidin-1 -yl)-2-carboxyvinyl)benzyl)- 6,7-dihvdroxyisoquinolin-2-ium 91 (500)
[0430] Under inert atmosphere, compound 90 (0.024 g, 0.027 mmol, 1 .0 equiv) is dissolved in anhydrous DCM (0.55 mL, 0.050 M). At 0 °C, TFA (0.55 mL, 0.050 M) is added dropwise and the resulting pale yellow solution is stirred at 0 °C for 20 minutes and then at rt for 1 .5 hours. Solvents are evaporated under reduced pressure and the residue is triturated twice in diethyl ether. The desired product 91 (500) is obtained as a white solid (0.014 g, 0.021 mmol, 77% yield).
[0431] 1H NMR (500 MHz, Acetone-de + D2O + 1 % TFA) d (ppm) 9.59 (s, 1 H), 8.44 (dd, 1 H, J = 7.0 Hz, J = 1 .5 Hz), 8.14 (d, 1 H, J = 7.0 Hz), 7.87-7.79 (m, 2H), 7.71 (s, 1 H), 7.62-7.54 (m, 2H), 7.55 (s, 1 H), 7.50 (s, 1 H), 7.03 (s, 1 H), 5.95 (s, 2H), 5.16 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 4.00 (dd, 1 H, J = 6.0 Hz, J = 5.0 Hz), 3.91 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 1 .52 (s, 3H), 1 .51 (s, 3H). m/z = 661 .5 [M]+, 331 .4 [M+H]2+. Example 7: Preparation of Exemplary Carbapenems of Formula I: Compound 484
Figure imgf000183_0001
A. Preparation of (5R6S)-4-nitrobenzyl 3-(4-(hvdroxymethyl)phenyl)-7-oxo-6-((R)- 1 -((triethylsilyl)oxy)ethyl)-1 -azabicvclo[3.2.01heot-2-ene-2-carboxylate 94
[0432] Under nitrogen atmosphere, (5R,6S)-4-nitrobenzyl 6-((R)-1 -hydroxyethyl)- 3,7-dioxo-1 -azabicyclo[3.2.0]heptane-2-carboxylate 92 (from Ark Pharm) (0.542 g, 1 .56 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (5.5 ml_, 0.28 M). At -78 °C, triethylamine (0.208 ml_, 1.50 mmol, 0.960 equiv) is added, followed by dropwise addition of trifluoromethanesulfonic anhydride (from Sigma Aldrich) (0.252 ml_, 1 .50 mmol, 0.960 equiv). The resulting reaction mixture is stirred at -78 °C for 0.5 hour and then triethylamine (0.239 ml_, 1 .71 mmol, 1 .10 equiv) is added, followed by dropwise addition of triethylsilyl trifluoromethanesulfonate (0.39 ml_, 1 .71 mmol, 1 .1 equiv). The resulting mixture is stirred at -78 °C for 1 hour. A solution of (4-(hydroxymethyl)- phenyl)boronic acid 93 (from AK Scientific) (0.237 g, 1 .56 mmol, 1 .00 equiv) in anhydrous DMF (3.5 ml_, 0.45 M) is added to the previous mixture, followed by tris(dibenzylideneacetone)-dipalladium(0) Pd2(dba)3 (0.029 g, 0.031 mmol, 0.020 equiv) and 6 M aqueous potassium hydroxide (0.78 ml_, 4.7 mmol, 3.0 equiv). The resulting reaction mixture is warmed to 30 °C and stirred at this temperature for 1 .5 hours. The reaction mixture is cooled to rt and is diluted in AcOEt (10 ml_). This mixture is filtered to remove any solids, and additional AcOEt (15 ml_) is added. The filtrate is washed with water (15 ml_) and the aqueous wash is extracted with AcOEt (6 ml_). The combined organic extracts are dried over sodium sulfate, filtered and then concentrated under reduced pressure. The residue is purified by column chromatography on silica gel (Biotage SNAP, 50 g) eluting with 0-50% AcOEt/DCM. A second column chromatography is run on fractions containing the product (Biotage SNAP, 50 g) eluting with 0-50% AcOEt/DCM. The desired product 94 is obtained as pale yellow film (0.283 g, 0.512 mmol, 33% yield).
[0433] 1H NMR (500 MHz, CDC ) d (ppm) 8.22-8.15 (m, 2H), 7.53-7.47 (m, 2H), 7.38-7.32 (m, 2H), 5.38 (d, 1 H, J = 13.5 Hz), 5.21 (d, 1 H, J = 14.0 Hz), 4.72 (d, 2H, J = 6.0 Hz), 4.34-4.24 (m, 2H), 3.29 (dd, 1 H, J = 27.5 Hz, J = 18. 5 Hz), 3.25 (dd, 1 H, J = 6.5 Hz, J = 3.0 Hz), 3.18 (dd, 1 H, J = 18.0 Hz, J = 9.5 Hz), 1 .71 (t, 1 H, J = 5.5 Hz), 1 .32 (d, 3H, J = 6.0 Hz), 0.97 (t, 9H, J = 7.5 Hz), 0.63 (q, 6H, J = 8.0 Hz) m/z = 553.2 [M+H]+.
Figure imgf000184_0001
B. Preparation of 1 -(4-((5R6S)-2-(((4-nitrobenzyl)oxy)carbonyl)-7-oxo-6-((R)-1 - ((triethylsilyl)oxy)ethyl)-1 -azabicvclo[3.2.01hept-2-en-3-yl)benzyl)pyridin-1 -ium 95
[0434] Under inert atmosphere, compound 94 (0.130 g, 0.235 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (4.0 ml_, 0.059 M). At 0 °C, pyridine (0.080 ml_, 0.99 mmol, 4.2 equiv) is added, followed by dropwise addition of trifluoromethanesulfonic anhydride (0.079 ml_, 0.47 mmol, 2.0 equiv. The resulting reaction mixture is stirred at 0 °C for 1 hour and 10 minutes. The reaction mixture is diluted in DCM (10 ml_) and this organic layer is washed with water (2 x 6 ml_). The organic layer is dried over sodium sulfate, filtered and concentrated under reduced pressure. The desired product 95 is obtained as orange solid film (0.182 g, quantitative yield). This material is used without further purification in the next step.
[0435] 1H NMR (500 MHz, Acetone-de) d (ppm) 9.33-9.27 (m, 2H), 8.83-8.76 (m, 1 H), 8.35-8.29 (m, 2H), 8.22-8.16 (m, 2H), 7.70-7.65 (m, 2H), 7.63-7.54 (m, 4H), 6.10 (s, 2H), 5.41 (d, 1 H, J = 13.5 Hz), 5.27 (d, 1 H, J = 13.5 Hz), 4.40-4.30 (m, 2H), 3.58 (dd, 1 H, J = 18.0 Hz, J = 8.5 Hz), 3.48 (dd, 1 H, J = 4.5 Hz, J = 3.0 Hz), 3.27 (dd, 1 H, J = 18.0 Hz, J = 10.5 Hz), 1 .28 (d, 3H, J = 6.0 Hz), 0.97 (t, 9H, J = 8.0 Hz), 0.64 (q, 6H, J = 7.5 Hz) m/z = 614 [M]+.
Figure imgf000185_0001
C. Preparation of (1 -(4-((5R6S)-6-((F?)-1 -hvdroxyethyl)-2-(((4-nitrobenzyl)- oxy)carbonyl)-7-oxo-1 -azabicvclo[3.2.01hept-2-en-3-yl)benzyl)pyridin-1 -ium 96
[0436] Compound 95 (0.123 g, 0.161 mmol, 1 .00 equiv) is dissolved in THF (15 mL, 0.01 1 M). Water (3 mL, 0.05 M) is then added. The resulting solution is treated with 1 M aqueous HCI until the pH of the reaction is approximately 2-3. The reaction mixture is stirred at rt for 3.5 hours. The pH of the reaction mixture is adjusted to 7 by addition of a dilute aqueous solution of NaHCCb. Solvents are evaporated under reduced pressure. The desired product 96 is obtained as orange solid film (0.1 17 g, quantitative yield). This material is lyophilized and used without further purification in the next step.
[0437] 1H NMR (500 MHz, Acetone-de) d (ppm) 9.27-9.21 (m, 2H), 8.74-8.67 (m, 1 H), 8.28-8.21 (m, 2H), 8.18-8.13 (m, 2H), 7.64-7.59 (m, 2H), 7.60-7.55 (m, 2H), 7.54- 7.49 (m 2H), 6.02 (s, 2H), 5.38 (d, 1 H, J = 14.5 Hz), 5.26 (d, 1 H, J = 13.5 Hz), 4.36-4.29 (m, 1 H), 4.15-4.07 (m, 1 H), 3.52 (dd, 1 H, J = 18.0 Hz, J = 8.0 Hz), 3.40 (dd, 1 H, J = 6.5 Hz, J = 2.5 Hz), 3.24 (dd, 1 H, J = 18.0 Hz, J = 9.5 Hz), 1 .25 (d, 3H, J = 6.0 Hz) m/z = 500 [M]+.
Figure imgf000185_0002
D. Preparation of (1 -(4-((5F?,6S)-2-carboxy-6-((F?)-1 -hvdroxyethyl)-7-oxo-1 - azabicvclo[3.2.01hept-2-en-3-yl)benzyl)pyridin-1 -ium 97 (484)
[0438] Under nitrogen atmosphere, compound 96 (0.042 g, 0.084 mmol, 1 .0 equiv) is dissolved in acetonitrile (0.60 ml_, 0.14 M). At rt, a phosphate buffer (0.5 M, pH 6.4, 1 .6 ml_) is added, followed by zinc powder (0.319 g, 4.88 mmol, 58.0 equiv). The suspension is vigorously stirred at rt for 1 hour and 10 minutes. Celite is added to the reaction mixture and the latter is filtered through a pad of celite. The solids are washed with a 1 : 1 (v/v) ACN/H2O mixture (3 x 5 ml_). The filtrate is concentrated under reduced pressure. The residue is purified by reverse phase column chromatography (Biotage C18, 12 g) eluting with 0-100% ACN/H2O. Fractions containing the desired product are combined and lyophilized. The desired product 97 (484) is obtained as a white solid (0.004 g, 0.01 mmol, 13% yield).
[0439] 1H NMR (500 MHz, D2O) d (ppm) 8.91 -8.84 (m, 2H), 8.56-8.50 (m, 1 H), 8.07-8.00 (m, 2H), 7.43-7.37 (m, 4H), 5.78 (s, 2H), 4.32-4.25 (m, 1 H), 4.26-4.20 (m, 1 H), 3.49 (dd, 1 H, J = 5.5 Hz), 3.41 (dd, 1 H, J = 17.0 Hz, J = 8.5 Hz), 3.06 (dd, 1 H, J = 17.0 Hz, J = 9.5 Hz), 1 .29 (d, 3H, J = 6.5 Hz) m/z =365.1 [M]+.
Example 8: Preparation of Exemplary Carbapenems of Formula I: Compound 488
Figure imgf000186_0001
A. Preparation of (5F?,6S)-4-nitrobenzyl 6-((F?)-1 -((fe/f-butyldimethylsilyl)oxy)ethyl)- 3-((E)-4-(hvdroxymethyl)styryl)-7-oxo-1 -azabicvclo[3.2.01hept-2-ene-2-carboxylate 99
[0440] Under nitrogen atmosphere, (5R,6S)-4-nitrobenzyl 6-((R)-1 -hydroxyethyl)- 3,7-dioxo-1 -azabicyclo[3.2.0]heptane-2-carboxylate 92 (from Ark Pharm) (4.10 g, 1 1 .8 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (39 ml_, 0.30 M). At - 78 °C, triethylamine (1 .58 ml_, 1 1 .3 mmol, 0.960 equiv) is added, followed by dropwise addition of trifluoromethanesulfonic anhydride (from Sigma Aldrich) (1 .90 ml_, 1 1 .3 mmol, 0.960 equiv). The resulting reaction mixture is stirred at -78 °C for 0.5 hour and then triethylamine (1 .80 ml_, 13.0 mmol, 1 .10 equiv) is added, followed by dropwise addition of fe/f-butyldimethylsilyl trifluoromethanesulfonate (2.97 ml_, 13.0 mmol, 1 .10 equiv). The resulting mixture is stirred at -78 °C for 0.5 hour. A solution of (E)-(4-(2-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)vinyl)phenyl)methanol 98 (from AK Scientific) (3.06 g, 1 1 .8 mmol, 1 .00 equiv) in anhydrous DMF (24 ml_, 0.50 M) is then added to the mixture, followed by addition of tris(dibenzylideneacetone)dipalladium(0) Pd2(dba)3 (0.22 g, 0.24 mmol, 0.020 equiv) and 6 M aqueous potassium hydroxide (5.9 ml_, 35 mmol, 3.0 equiv). The resulting reaction mixture is warmed to 30 °C and stirred at this temperature for 1 hour. The reaction mixture is cooled to rt and is diluted in AcOEt (100 ml_). This organic layer is washed with water (100 ml_) and the aqueous wash is extracted with AcOEt (100 ml_). The combined organic extracts are dried over magnesium sulfate, filtered and then concentrated under reduced pressure. The residue is purified by column chromatography on silica gel eluting with 0-100% AcOEt/hexanes. The desired product 99 is obtained as pale yellow solid (4.03 g, 6.96 mmol, 59% yield).
[0441] 1H NMR (500 MHz, CDCIs) d (ppm) 8.25-8.20 (m, 2H), 7.91 (d, 1 H, J = 16.0 Hz), 7.71 -7.66 (m, 2H), 7.48-7.44 (m, 2H), 7.36-7.31 (m, 2H), 6.70 (d, 1 H, J = 17.0 Hz), 5.48 (d, 1 H, J = 13.5 Hz), 5.30 (d, 1 H, J = 14.0 Hz), 4.31 -4.20 (m, 2H), 3.22 (dd, 1 H, J = 17.5 Hz, J = 9.5 Hz), 3.18 (dd, 1 H, J = 5.5 Hz, J = 3.0 Hz), 3.10 (dd, 1 H, J = 17.5 Hz, J = 9.0 Hz), 1 .29 (d, 3H, J = 6.0 Hz), 0.89 (s, 9H), 0.10 (s, 6H).
Figure imgf000187_0001
B. Preparation of (5F?,6S)-4-nitrobenzyl 6-((F?)-1 -((fe/f-butyldimethylsilyl)oxy)ethyl)- 3-((E)-4-(chloromethyl)styryl)-7-oxo-1 -azabicvclo[3.2.01hept-2-ene-2-carboxylate 100
[0442] Under inert atmosphere, compound 99 (5.21 g, 9.01 mmol, 1 .00 equiv) is dissolved in anhydrous DCM (90 ml_, 0.10 M). At rt, triethylamine (1 .82 ml_, 13.5 mmol, 1 .50 equiv) is added, followed by dropwise addition of methanesulfonyl chloride (1 .05 ml_, 13.5 mmol, 1 .50 equiv). DMAP (0.1 1 g, 0.90 mmol, 0.10 equiv) is added and the resulting reaction mixture is stirred at rt for 18 hours. Brine (50 ml_) is added and the biphasic mixture is transferred to a separatory funnel. The aqueous layer is extracted with DCM (2 x 50 ml_). The organic extracts are combined and dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue is purified by column chromatography on silica gel eluting with 0-25% AcOEt/hexanes gradient. The desired product 100 is obtained (4.32 g, 7.23 mmol, 80% yield).
[0443] 1H NMR (500 MHz, Acetone-de) d (ppm) 8.25-8.19 (m, 2H), 7.93 (d, 1 H, J = 16.0 Hz), 7.72-7.67 (m, 2H), 7.48-7.43 (m, 2H), 7.38-7.32 (m, 2H), 6.69 (d, 1 H, J = 16.0 Hz), 5.48 (d, 1 H, J = 14.0 Hz), 5.30 (d, 1 H, J = 13.5 Hz), 4.58 (s, 2H), 4.31 -4.22 (m, 2H), 3.26-3.17 (m, 2H), 3.15-3.07 (m, 1 H), 1 .28 (d, 3H, J = 5.5 Hz), 0.87 (s, 9H), 0.09 (s, 6H).
Figure imgf000188_0001
C. Preparation of (5F?,6S)-4-nitrobenzyl 3-((E)-4-(chloromethyl)styryl)-6-( -1 - hvdroxyethyl)-7-oxo-1 -azabicvclo[3.2.01hept-2-ene-2-carboxylate 101
Figure imgf000188_0002
[0444] Under nitrogen atmosphere, compound 100 (4.32 g, 7.23 mmol, 1 .00 equiv) is dissolved in anhydrous THF (72 ml_, 0.10 M). Acetic acid (1.9 ml_, 33 mmol, 4.5 equiv) is added at rt, followed by TBAF (1 .0 M solution in THF, 21 .7 ml_, 21 .7 mmol, 3.00 equiv). The resulting solution is stirred at rt for 19 hours. Saturated aqueous sodium bicarbonate (50 ml_) is added to the reaction mixture, followed by brine (50 ml_). The biphasic mixture is transferred into a separatory funnel and the mixture is extracted with AcOEt (2 x 50 mL). The organic layer is dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue is purified by column chromatography on silica gel eluting with 0-100% AcOEt/hexanes gradient. The desired product 101 is obtained as pale yellow solid (0.597 g, 1 .24 mmol, 17% yield).
[0445] 1H NMR (500 MHz, CDC ) d (ppm) 8.26-8.23 (m, 2H), 7.92 (d, 1 H, J = 16.0 Hz), 7.72-7.67 (m, 2H), 7.47-7.43 (m, 2H), 7.37-7.34 (m, 2H), 6.70 (d, 1 H, J = 16.0 Hz), 5.54 (d, 1 H, J = 13.5 Hz), 5.30 (d, 1 H, J = 13.5 Hz), 4.58 (s, 2H), 4.33-4.24 (m, 2H), 3.32-3.22 (m, 2H), 3.13 (dd, 1 H, J = 17.0 Hz, J = 8.5 Hz), 1 .69 (d, 1 H, J = 5.0 Hz), 1 .39 (d, 3H, J = 6.5 Hz).
Figure imgf000189_0001
D. Preparation of 1 -(4-((E)-2-((5R6S)-6-((R)-1 -hydroxyethyl)-2-(((4-nitrobenzyl)- oxy)carbonyl)-7-oxo-1 -azabicyclo[3.2.01hept-2-en-3-yl)vinyl)benzyl)pyridin-1 -ium 102
[0446] Under nitrogen atmosphere, compound 101 (0.060 g, 0.12 mmol, 1 .0 equiv) is dissolved in anhydrous DMF (1 .2 mL, 0.10 M). At rt, pyridine (0.020 mL, 0.25 mmol, 2.0 equiv) is added, followed by addition of potassium iodide (0.021 g, 0.12 mmol, 1 .0 equiv). The resulting mixture is stirred at 30 °C for 7 hours. The product is precipitated out by addition of diethyl ether and vigorous stirring for 5 minutes at rt. The mixture is sonicated and the diethyl ether layer is removed. The solid is then triturated twice in water. The resulting solid is sonicated in diethyl ether. The diethyl ether layer is removed. Remaining traces of solvent are removed by rotatory evaporation and by drying under high vacuum. The desired product 102 is obtained as brown solid (0.046 g, 0.087 mmol, 73% yield).
[0447] m/z =526.3 [M]+.
Figure imgf000190_0001
102 103
E. Preparation of 1 -(4-((E)-2-((5 ?,6S)-2-carboxy-6-(( ?)-1 -hvdroxyethyl)-7-oxo-1 - azabicvclo[3.2.01hept-2-en-3-yl)vinyl)benzyl)pyridin-1 -ium 103 (488)
[0448] Compound 102 (0.047 g, 0.070 mmol, 1.0 equiv) is dissolved in THF (1.4 ml_, 0.050M). At rt, a phosphate buffer (0.5 M, pH 6.4, 1.4 ml_) is added, followed by addition of zinc powder (0.133 g, 2.03 mmol, 29.0 equiv). The suspension is vigorously stirred at rt for 1.5 hours. Celite is added to the reaction mixture and the latter is filtered through a pad of celite. The solids are washed with a 1 : 1 v/v THF/H2O mixture. The filtrate is washed with DCM and the aqueous extract is lyophilized. The desired product is extracted from the solid by trituration in MeOH (3 x 5 ml_). The filtrate is concentrated under reduced pressure. The remaining phosphate is removed by solubilizing the product in a minimum of water and passing the solution through an anion exchange resin (IRA-400(CI)) column. The product is eluted with water. Fractions containing the desired product are combined and lyophilized. The desired product 103 (488) is obtained as a yellow solid (0.023 g, 0.059 mmol, 84% yield).
[0449] 1H NMR (300 MHz, D2O) d (ppm) 8.85-8.79 (m, 2H), 8.52-8.44 (m, 1 H), 8.03-7.95 (m, 2H), 7.73 (d, 1 H, J = 16.2 Hz), 7.58-7.51 (m, 2H), 7.40-7.32 (m, 2H), 6.65 (d, 1 H, J = 16.5 Hz), 5.73 (s, 2H), 4.21 -4.07 (m, 2H), 3.34 (dd, 1 H, J = 6.0 Hz, J = 3.0 Hz), 3.09 (dd, 1 H, J = 9.3 Hz, J = 6.3 Hz), 1.23 (d, 3H, J = 6.3 Hz) m/z =391.2 [M]+.
Example 9: Antibiotic Activity
[0450] The MIC of compounds having Formula I were measured across 12 different organisms to evaluate the potential antibacterial activity for the compounds of the present disclosure. In this study, the following compounds were assayed and determined to have MIC values that inhibit the growth of bacteria at relatively low concentrations: 501 , 502, 503, 504, 482, 487, 505, 506, and 507. Methods
[0451] Measurement of Minimum Inhibitory Concentrations (MIC). Sterile technique is used throughout these experiments. Suspensions of organisms are removed from storage (-80°C) and partially thawed. Partially thawed organisms (1 mI_) were then inoculated into 3 ml_ sterile lysogeny broth in a sterile culture tube and capped, then cultured at 37°C with shaking for 4-5 hours until the optical density at 600 nm (ODeoo) is greater than 0.1 on a single beam spectrophotometer. The concentration of organisms is adjusted to 0.08 - 0.1 OD6oo (0.5 McFarland units) by dilution with sterile cation-adjusted Meuller-Hinton (CAMH) broth. The suspensions are used within 30 minutes of preparation.
[0452] Test Compound Stock Solution. A stock solution is made of each test compound (by measuring 6.4 mg of the compound on an analytical balance and dissolving it in DMSO (1 ml_) to get a transparent solution. The solutions are stored at -20°C and thawed prior to use.
[0453] MIC Determination. Sterile CAMH broth (200 pL) containing 20 pM Apo- Transferrin is pipetted into the first row of 8 columns of a sterile, round-bottomed, transparent, 96-well microplate using an 8-channel pipette equipped with sterile tips. One hundred pL is pipetted into the wells of the remaining 1 1 rows. To each well of the first row is pipetted 2.0 pL of the 6.4 mg/mL stock test compound stock solution to give 1 : 100 dilutions and a final concentration of 64 pg/mL and mixed.
[0454] With an 8-channel pipette equipped with sterile tips, 100 pL of the first row solution is transferred to the second row and mixed. The process is repeated for the second to third rows and continued through row 1 1 . One hundred pL from row 1 1 is removed and discarded so that all 12 rows contained 100 pL. Row 12 contained no compound and rows 1 - 1 1 contained 64, 32, 16, 8, 4, 2, 1 , 0.5, 0.25, 0.125, 0.6 pg/mL of a given compound.
[0455] Inoculant suspensions (0.5 pL) are added to the 12 wells of each column. The plates are then covered and incubated at 37°C overnight for 18 hours. For each organism the MIC is determined by identifying the transparent well with the lowest test compound concentration.
[0456] Strain with the following beta-lactamases were employed: [0457] TEM-26 (Group 2be beta-lactamase, hydrolyzes extended spectrum cephalosporins and monobactams), [28];
[0458] SHV-1 (Class A, Group 2b beta-lactamase, hydrolyzes penicillins and early cephalosporins) [29];
[0459] DHA-1 (beta-lactamase, intermediate or resistant to all penicillins, alone or in combination with beta-lactamase inhibitors, and to cephalothin, cefuroxime, cefoxitin, ceftazidime, aztreonam, and cefotaxime) [30, 31 ];
[0460] CTX-M-14 and CTX-M-15 (Class A, Group 2be beta-lactamases, hydrolyzes extended spectrum cephalosporins and monobactams) [32];
[0461] KPC-2 and KPC-3 (Class A, Group 2f, carbapenemases) [33, 34];
[0462] NDM-1 (Group 3a carbapenemase, broad spectrum divalent Zn++ metallo- Beta-Lactamase) [35, 36];
[0463] VIM-2 (Group 3a carbapenemase, metallo-beta-lactamase) [37]; and
[0464] IMP-1 (Group 3a carbapenemase, metallo-beta-Lactamase).
[0465] MIC results for E. coli J58 isogenic strains producing ESBL (Extended Spectrum beta-Lactamases) and other beta-lactamases [26, 27] are provided.
[0466] The gene sequence for each beta-lactamase enzyme is synthesized and inserted into the pBC-SK(+) plasmid by GenScript, a commercial service. E.coli J53 is transformed with each plasmid via electroporation and resistant transformants are selected on agar plates containing 50 pg/mL ampicillin. The selected transformants are stored at -80 degrees C in 20% glycerol.
Results
[0467] MIC (pg/mL) results are provided for exemplary compounds of Formula I in Tables 1 -6. Organisms for which MICs are measured are known in the art, can be prepared by well-known methods from available starting materials are available from commercial sources or are clinical isolates. Tables 1 -6 show that the compounds of Formula I (e.g., 501 , 502, 503, 504, 482, 487, 505, 506, and 507) exhibit MIC values that indicate the compounds can inhibit bacterial growth at relatively low concentrations and across vast number of difference organisms. Conclusion
[0468] The results from this study demonstrate the potential for compounds of Formula I to serve as antimicrobial agents by inhibiting bacterial growth at relatively low concentrations.
Example 10: Bactericidal Activity of GL-332
[0469] The MIC is defined as the lowest concentration of an antimicrobial ingredient or agent that is bacteriostatic (prevents the visible growth of bacteria). MICs are used to evaluate the antimicrobial efficacy of various compounds by measuring the effect of decreasing concentrations of the antibacterial agent (e.g., GL332) over a defined period in terms of inhibition of microbial population growth. The MBC is the lowest concentration of an antibacterial agent required to kill a bacterium over a fixed. The MBC is complementary to the MIC; whereas the MIC test demonstrates the lowest level of antimicrobial agent that greatly inhibits growth, the MBC demonstrates the lowest level of antimicrobial agent resulting in microbial death.
[0470] When the MBC to MIC ratio is < 4, the antimicrobial agent is considered bactericidal. Conversely, when the MBC to MIC ratio is > 8, the antimicrobial agent is considered bacteriostatic. Bactericidal agents kill bacteria, rather than just prevent the growth of bacteria and are therefore, considered to be more potent than bacteriostatic agents.
[0471] In this study, the spectrum of activity was determined for GL-332 by conducting minimal inhibitory concentration (MIC) testing against a range of Gram positive/negative bacteria that included aerobes and anaerobes in medium depleted of iron through treatment with Chelex; in some instances, testing was performed in untreated medium as well. In addition, the minimum bactericidal concentration (MBC) was determined for GL-332 against a subset of these organisms, which was performed in medium that had been depleted of iron following treatment with Chelex.
[0472] The results from this study show that GL332 has bactericidal activity against several isolates of E. coli, K. pneumoniae and P. aeruginosa.
Methods
[0473] Test Media: Broth microdilution testing of aerobic bacteria was conducted in cation-adjusted Mueller Hinton broth (MHBII). For testing Streptococci, Listeria, Campylobacter and Corynebacterium, lysed horse blood (LHB) was added to MHBII broth at a final concentration of 3%. For testing Haemophilus, Haemophilus test medium (HTM) was used. For testing Neisseria, GC media was used.
[0474] For broth microdilution MIC testing of GL-332 only, iron-depleted medium was used. Chelex 100 resin was added (100 g/liter of autoclaved medium) and stirred for 2 hr at room temperature to remove cations from the medium. Chelex was removed by passing medium through a 0.2-micron filter. The medium was supplemented with CaCl2 to 22.5 mg/liter (range, 20 to 25 mg/liter), MgCte to 1 1 .25 mg/liter (range, 10 to 12.5 mg/liter), and ZnSC to 10 mM (0.56 mg/liter; range 0.5 to 1 .0 mg/liter). The pH of the iron-depleted medium was adjusted to 7.3 using hydrochloric acid. The medium was then sterilized by passing through a 0.2-micron filter, followed by passing through a second 0.2-micron filter. Where LHB was used, the blood was added after all filtering.
[0475] Broth Microdilution MIC Assay: Broth microdilution susceptibility testing in 96-well microplates was conducted. Automated liquid handlers were used to conduct serial dilutions and liquid transfers, including the Biomek 2000, and the Biomek FX (Beckman Coulter, Fullerton, CA). All wells in columns 2 through 12 of a standard 96- well microdilution plate (Costar 3795) were filled with 150 pL of the appropriate diluent. Then, 300 pL of the tested agents were added to the wells of column 1 of the plates at 100X the highest final concentration to be tested. Serial two-fold dilutions were made across the rows through column 1 1 using the Biomek 2000. The wells of column 12 contained no drug and served as the growth control wells. This plate served as the “mother plate” from which MIC assay plates or“daughter plates” were made.
[0476] The daughter plates were loaded with 188 pL per well of the appropriate medium by hand. The daughter plates were created using the Biomek FX which transferred 2 pL of drug solution from each well of a mother plate to each corresponding well of the daughter plate in a single step. A standardized inoculum of each test organism was prepared per CLSI methods (1 ) to equal a 0.5 McFarland standard, followed by a dilution of 1 :20. The plates were then inoculated with 10 pL of the diluted inoculum using the Biomek 2000 from low to high drug concentration, resulting in a final concentration of approximately 5 x 105 CFU/mL per well. An un-inoculated plate was incubated in parallel for each medium for the purpose of assessing solubility of the drug in the test media. [0477] Minimum Bactericidal Concentration (MBC) Assay. MBC values were determined in accordance with CLSI guidelines [38] The viable count of each tested inoculum was determined immediately post-inoculation of the 96-well broth microdilution test plates by removing a 0.3 mL volume from the McFarland 0.5 standard, preparing serial 10-fold dilutions in 0.27 mL of sterile saline, and spotting 10 pi aliquots across the top of a trypticase soy agar plate. The plate was then tilted at a 45°- 90° angle to allow the 10 pL aliquot to track across the agar surface to the opposite side of the plate. The plates were laid flat, allowed to dry at room temperature, then inverted and incubated at 35°C for approximately 24 hr. Colonies were manually counted to determine the viable inoculum count in CFU/mL in each well of the corresponding MIC plate.
[0478] Following incubation of the 96-well plates and recording the broth microdilution MIC values, duplicate 10 pL aliquots from the MIC well and three wells above the MIC were spotted and dragged across the surface of a TSAII plate. Plates were allowed to dry before inverting and incubation overnight at 35°C. The number of colonies per plate were counted manually. The sums of the counts for the two spotted aliquots were compared to the values in the appropriate table of rejection values [38] These values were based upon the cell density of the inoculum and the target viable count reduction of 99.9%; if the sum of the colonies was less than or equal to the value in the table, the concentration of drug in the sampled well was considered to be bactericidal [38] The MBC was then defined as the lowest concentration of agent to demonstrate a bactericidal effect.
Results
[0479] In this study, GL332 was observed to have bactericidal activity against several isolates of E. coli, K. pneumoniae and P. aeruginosa. The MIC and MBC values for GL332 per organism are enumerated in Table 7. As stated above, MBC:MIC ratios < 4 are commonly observed for bactericidal agents and are thus considered indicative of bactericidal activity as opposed to ratios > 8 which are common among bacteriostatic agents.
[0480] As shown in Table 7, MBC:MIC ratios for each organism is < 4, indicating that GL332 has bactericidal activity. For example, the MIC:MBC ratio for E. Coli (source MMX 0102 ATCC 25922) was 1 (i.e., MIC is 0.06 mg/L and MBC is 0.06 mg/L), the MIC:MBC ratio for K. pneumoniae (source MMX 4972 ATCC 43816) was also 1 (i.e. , MIC of 0.03 mg/L and MBC of 0.03 mg/L), and lastly, the MIC:MBC ratio for P. aeruginosa PA01 (source Gladius) was also 1 (i.e., MIC of 0.5 mg/L and MBC of 0.5 mg/L).
Table 7. MIC and MBC Values for GL332
Figure imgf000196_0001
Figure imgf000197_0001
PC = unspecified KPC class A serine carbapenemase.; ESBL = extended spectrum b lactamase; CTX-M-14 is an extended spectrum b lactamase; QC = quality control strain; ColR = colistin resistant; All MICs determined in iron depleted broth.
Conclusion
[0481] Accordingly, the results from this study show GL332 has very potent bactericidal activity against each of the tested organisms E. coli, K. pneumoniae and P. aeruginosa. In particular, the MIC:MBC values for E. coli, K. pneumoniae and P. aeruginosa were <4, indicating that GL-332 exhibits strong bactericidal activity rather than the lesser bacteriostatic activity.
Example 11 : Serum Shift Data for GL332
[0482] Plasma protein binding (PPB) is an important consideration for drug discovery as it influences the free fraction of drug available to bind to its target. Human serum albumin (HAS) is an abundant extracellular protein found in the blood plasma and tissue fluids. HAS is able to bind and then transport a number of endogenous compounds found in the body. However, HAS also absorbs a significant amount of drug in plasma and tissue fluids. The pharmacological activity of a drug is, in part, determined by the concentration of unbound concentration of the drug, not the total concentration of the blood in the plasma and tissue fluids. Accordingly, drugs that show a high affinity for HAS require higher dosing in order to achieve a therapeutic effect.
[0483] In this study, GL-332 was evaluated for in vitro activity against a panel of Gram-negative aerobic bacteria in the presence and absence of mouse serum to determine the effects of plasma proteins on the MIC values for GL-332.
[0484] The results from this study demonstrate that the MIC value of GL-332 decreases in the presence of mouse serum as compared to in the absence of mouse serum, suggesting that GL-332 is not highly bound by the plasma proteins. These results are significant as they suggest there is synergy between the plasma proteins and GL-332 and that a high dosing of GL-332 is not required to achieve a desired therapeutic effect.
Methods
[0485] The activity of GL-332 was evaluated in the presence and absence of mouse serum by conducting MIC testing against a range of Gram-negative aerobic bacteria. Isolates were chosen based on in vitro activity of GL-332. Studies were conducted following guidelines published by the Clinical and Laboratory Standards Institute using broth microdilution; concentrations of mouse serum used during testing were 0, 20 and 50%. Imipenem, amikacin, levofloxacin, and meropenem were the comparator agents in this study.
Results
[0486] Table 8 shows the results of evaluating the activity of GL-332 against several Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa isolates in the presence and absence of mouse serum. All testing of GL-332 against these organisms was performed in iron-depleted medium (testing of comparators was performed in MBH II). All QC results were within published CLSI ranges for these drugs, thus validating this assay.
Table 8: In vitro activity of GL-332 against Gram-negative aerobic bacteria in the presence and absence of mouse serum
Figure imgf000198_0001
ESBL, extended-spectrum b-lactamase producing; NG, no growth. IMI, imipenem; AMK, amikacin, LVX, levofloxacin; MER, meropenem.
[0487] As show in Table 8, against three E. coli isolates, including one extended- spectrum b-lactamase-producing (ESBL+) strain, the activity of GL-332 was repeatedly observed to improve in the presence of 20% mouse serum, up to 16-fold higher in the case of isolate MMX 8427. Against one E. coli isolate the activity of GL-332 decreased in the presence of 50% mouse serum (MMX 9340), but was improved four-fold against another (MMX 8427; the third isolate did not grow under those conditions).
[0488] Two of the K. pneumoniae isolates tested were ESBL+ (MMX 9028 and MMX 9029) and the third was colistin-resistant (MMX 8403); against these, GL-332 had MIC values of 0.06, 0.25 and 0.5 pg/mL. There was little impact of mouse serum on the activity of GL-332 against K. pneumoniae MMX 9028; 20% mouse serum improved the activity of GL-332 four to eight-fold but 50% mouse serum had no impact on activity against the remaining two K. pneumoniae isolates.
[0489] The MIC values for GL-332 against P. aeruginosa MMX 7748 and MMX 7753 were 0.25 and 1 pg/mL, respectively. For P. aeruginosa MMX 7753, these values were not changed in either 20% or 50% mouse serum, though in the case of P. aeruginosa MMX 7748 in the presence of 50% mouse serum a four-fold increase in GL- 332 MIC was observed.
[0490] Lastly, the activities of imipenem, amikacin, levofloxacin and meropenem were either not impacted or negatively impacted by mouse serum in a strain-specific fashion. The one exception was amikacin against E. coli MMX 8424, where the activity was improved in the presence of serum.
Conclusion
[0491] The results from this study showed that in the presence of 20% mouse serum GL-332 was observed to demonstrate improved activity against E. coli and K. pneumoniae isolates generally, whereas this concentration of mouse serum had no effect on the activity of GL-332 against the P. aeruginosa isolates tested. Further, in presence of 50% mouse serum had either less of a positive impact or a negative impact (as in the case of E. coli MMX 9340 and P. aeruginosa MMX 7748) on the activity of GL-332, suggesting that the effects of higher serum concentrations are strain-specific. Example 12: GL332 Mouse PK & Comparators
[0492] The objective of this study was to determine the pharmacokinetics (PK) of GL-332 in mice and compare the efficacy to the comparator antibiotics, ceftazidime and ceftriaxone.
Methods
[0493] Sample Preparation for GL-332. The plasma samples were prepared by first adding a 5 mI of the GL-332, calibration standard, quality control, dilute quality control, single blank, and double blank samples to a 1 .5 mL tube. Each sample (except the double blank) was then quenched with 50 pL of an internal standard (6 in 1 internal standard in acetonitrile (Labetalol & tolbutamide & Verapamil & dexamethasone & glyburide & Celecoxib 100 ng/mL for each)) and vortexed for at least 15 seconds and then centrifuged from 15 min at 12,000 g at 4°C. 25 pL of the supernatant was then transferred to a clean 96-well plate a reconstituted with 25 pL of water, vortexed for 10 min at 800 rpm, and lastly, centrifuged for 5 min at 3,220 g at 4°C. 10 pL of the supernatant was then injected for LC-MS/MS analysis.
[0494] Sample Preparation for Cefrazidime. The plasma samples were prepared by first adding a 5 mI of ceftazidime, calibration standard, quality control, dilute quality control, single blank, and double blank samples to a 1 .5 mL tube. Each sample (except the double blank) was then quenched with 50 mί of an internal standard (6 in 1 internal standard in acetonitrile (Labetalol & tolbutamide & Verapamil & dexamethasone & glyburide & Celecoxib 100 ng/mL for each)) and vortexed for at least 15 seconds and then centrifuged from 15 min at 12,000 g at 4°C. 25 mί of the supernatant was then transferred to a clean 96-well plate a reconstituted with 25 mί of water, vortexed for 10 min at 800 rpm, and lastly, centrifuged for 5 min at 3,220 g at 4°C. 10 mί of the supernatant was then injected for LC-MS/MS analysis.
[0495] Sample Preparation for Ceftriaxone. The plasma samples were prepared by first adding a 5 mI of ceftazidime, calibration standard, quality control, dilute quality control, single blank, and double blank samples to a 1 .5 mL tube. Each sample (except the double blank) was then quenched with 50 mί of an internal standard (6 in 1 internal standard in methanol (Labetalol & tolbutamide & Verapamil & dexamethasone & glyburide & Celecoxib 100 ng/mL for each)) and vortexed for at least 15 seconds and then centrifuged from 15 min at 12,000 g at 4°C. 25 mI_ of the supernatant was then transferred to a clean 96-well plate a reconstituted with 25 mI_ of water, vortexed for 10 min at 800 rpm, and lastly, centrifuged for 5 min at 3,220 g at 4°C. 10 mI_ of the supernatant was then injected for LC-MS/MS analysis.
[0496] Dilution Procedure. To dilute each sample by a factor of 10, an aliquot of 2 mI_ of each sample was added to 18 mI_ plasma. To dilute each sample by a factor of 100, an aliquot of 2 pL sample was added to 18 pL blank plasma to obtain d/10 sample. An aliquot of 2 pL of d/10 sample was then added to 18 pL blank plasma 10 additional times to achieve a 100 fold dilution.
[0497] LC-MS/MS Analysis. The data were collected by LC-MS/MS using ACQUITY UPLC HSS T3 1.8 pm 2.1 c 50 mm column at temperature of 60°C and flow rate of 0.6 mL/min (positive, ESI). GL-332 was observed at a [M+H]+ m/z of 353.1/303.2, ceftazidime was obverse at a [M+H]+ m/z of 547.1/468.1 , and lastly, ceftriaxone was observed at [M+H]+ m/z 555.0/396.1 .
Results
[0498] The PK parameters for GL-332 and the comparators, ceftazidime, and ceftriaxone are enumerated in Table 9 and FIG. 1. As shown in FIG. 1 , GL-332 has pharmacokinetic profile similar to other clinically available cephalosporins with an early elimination rate similar to ceftazidime and a terminal elimination half-life similar to ceftriaxone.
[0499] Table 9 also indicates that GL-332 has similar, if not better, PK parameters as compared to ceftazidime and ceftriaxone. For example, the half-life (T1/2) is approximately 6-fold greater for GL-332 (1 .6±0.5 h) as compared to ceftazidime (0.28±0.02 h) and comparable to ceftriaxone (1 .48±0.03 h).
Table 9. PK Parameters for GL-332, Ceftazidime, and Ceftriaxone
Figure imgf000201_0001
Figure imgf000202_0001
Conclusion
[0500] The results from this study indicate that GL-332 behaves similarly to clinically approved cephalosporins, ceftazidime and ceftriaxone. For example, GL-332 has pharmacokinetic profile shows that GL-332 has an early elimination rate similar to ceftazidime and a terminal elimination half-life similar to ceftriaxone (FIG 1 ).
Example 13: Penicillin-Binding Proteins Inhibition Assay for GL-332
[0501] b-Lactam antibiotics act by binding to penicillin-binding proteins (PBPs) and disrupting peptidoglycan cross-linking during cell wall synthesis, resulting in bacterial lysis and cell death. One way to measure the effectiveness of an antibiotic (e.g., GL- 332) at inhibiting bacterial growth is to determine the half maximal inhibitory concentration (ICso) of the antibiotic. This quantitative measure indicates how much of a particular antibiotic is needed to inhibit bacterial growth.
[0502] The objective of this study was to assess the inhibitory efficacy of GL-332 on bacterial growth by determining the ICso value for PBP1 , PBP2, and PBP3. The results from this study indicate that GL-332 strongly inhibits PBP3 with an ICso value of 10 ng/mL and also inhibits PBP1 and PBP2, although not to the same degree, with an ICso value greater than 10 mg/L.
Methods
[0503] 1.5 mL of an exponential culture lysate (OD600=0.5) was resuspended in
50 pi PBS containing 100 pg/mL, 10 pg/mL, 1 pg/mL, 100 ng/mL, 10 ng/mL, 1 ng/mL, 100 pg/mL, 10 pg/mL, 1 pg/mL, 100 fg/mL, 10 fg/mL of GL-332, two references sample in PBS without antibiotic, and 1 reference sample with 10mg/mL penicillin G in PBS. After 30 min of incubation at room temperature, the lysate was pelleted at 21 ,000 g for 15 min at 4 °C, the pellet washed in PBS, and pellet resuspended in 50 mI PBS containing 5 pg/ml Bocillin-FL (15 mM) (0.1 % DMSO). After a 10 min incubation, the lysate was again pelleted at 21 ,000g for 15 min at 4 °C, washed with PBS, and resuspended in 100 pi PBS.
[0504] The protein concentration was measured using a NanoDrop 1000 Spectrophotometer (260 and 280 nm) and all samples were adjusted to 5 mg/ml by dilution with PBS. A 51 mI_ volume of proteome sample and 15 pL of 4x SDS-PAGE loading buffer were mixed and heated for 5 min at 90 °C. A 12-mI volume of sample was loaded onto a 10% SDS-PAGE gel. Fluorescent intensity of the BODIPY fluorophore measured using a Typhoon FLA 9500 scanner (GE Healthcare) at 526-nm (short-pass filter) detection for BODIPY-FL (Aex: 504 nm, Aem: 514 nm) and quantified with ImageJ, subtracting lane background and averaging the no antibiotic reference controls.
Results
[0505] FIG. 2 shows that PBP3 ICso inhibition curve of GL-332 for PBP3. As shown in FIG. 2, GL-332 exhibited a very low ICso of PBP3 at a concentration of 10 ng/mL. GL-332 also showed an ICso for PBP1 and PBP2 of greater than 10 mg/L (data not shown).
[0506] The effects of GL-332 on binding to PBP3 was assessed using electrophoresis on bacterial lysates as shown in FIG. 3. As demonstrated in FIG. 3, as the concentration of GL-332 is increased (10 fg/mL to 100 pg/mL), the fluorescent signal decreases for both replicate 1 and replicate 2. ( See FIG. 3, area of interest emphasized with a rectangle). The decrease in the fluorescent signal indicates that as the concentration of GL-332 is increased more PBP3 is bound by GL-332, quenching the fluorescent signal of PBP3. Importantly, this data indicates that at relatively low concentrations, GL-332 is able to bind to PBP3, which would in turn, inhibits bacterial growth.
Conclusion
[0507] The results from this study show that GL-332 binds to one or more penicillin-binding proteins, consistent with arresting bacterial cell-wall synthesis and inhibiting bacterial replication. This data suggests that GL-332 can be used to treat bacterial infections at relatively low concentrations with GL-332 inhibiting bacterial growth by binding to PBP3 at an ICso of 10 ng/mL. Example 14: Inhibitory Efficacy of GL-332 on Pseudomonas aeruginosa
[0508] This study objective was to evaluate the in vitro antimicrobial activity and in vivo efficacy of GL-332 against Pseudomonas aeruginosa ATCC 27853.
[0509] The minimum inhibitory concentration (MIC) value of GL-332 was determined following the guidelines of the Clinical Laboratory Standards Institute (CLSI), M07-A10, for the standard broth microdilution method. The MIC value of GL-332 was 0.5 pg/mL. Ciprofloxacin was used as the reference standard and had an MIC value was 0.25 pg/mL, consistent with the historical data.
Methods
[0510] The in vivo efficacy of GL-332 was tested in the P. aeruginosa ATCC 27853 peritoneal infection model with immune competent ICR mice. Test animals were infected intraperitoneally (IP) with P. aeruginosa ATCC 27853 at target density of 5-7 c 105 CFU/mouse. The actual inoculum count was 5.4 c 105 CFU/mouse. Test substance, GL-332 at 40 mg/kg, was administered intravenously (IV) once (QD) at 1 h after infection. GL-332 at 20 and 40 mg/kg was administered IV twice (BID) with a 12 h interval (q12h) at 1 and 13 h after infection for a total dose of 40 or 80 mg/kg. The reference agent, gentamicin at 10 mg/kg, was administered subcutaneously (SC) BID q12h at 1 and 13 h after infection. Animal mortality was observed for 7 consecutive days. Fisher’s exact test was used to assess the statistical significance in the survival of the treated animals compared to the vehicle control group on Day 7. The p value < 0.05 indicated a significant increase in the survival rates of the treated animals.
[0511] MIC Method. GL-332 was dissolved and 2-fold diluted in 100% DMSO for a total of 1 1 concentrations ranging from 6.4 to 0.006 mg/mL. A 4 pL aliquot (50-fold stock solution) of each dilution was added to 196 pL of test medium seeded with test organisms in wells of a 96-well polystyrene round bottom micro-well plate. The target inoculum was 2 - 8 x 105 colony forming units (CFU)/mL. The final volume was 200 pL in each well. Test concentrations were 128, 64, 32, 16, 8, 4, 2, 1 , 0.5, 0.25 and 0.125 pg/mL. Following 18 h incubation at 35-37°C without shaking, the test plates were visually examined, and each well was scored for growth or complete inhibition of growth. Each test substance was evaluated in duplicate. Vehicle-control and active reference ciprofloxacin were used as blank and positive controls, respectively. [0512] P. aeruginosa (ATCC 27853), Peritoneal Infection Model, LD90-100: Inoculum Preparation. The testing strain, P. aeruginosa (ATCC 27853), was obtained from a frozen working stock, thawed at room temperature. A 0.2 mL aliquot was inoculated into 20 mL TSB and incubated at 35-37°C with shaking (250 rpm) for 6 h. One mL of the 6 h culture was used to seed 99 mL TSB and incubated at 35-37°C with shaking at 250 rpm for 16 h. Cells in 20 mL culture were pelleted by centrifugation (3,500 x g) for 15 minutes, then resuspended in 10 mL cold PBS (>1 c 109 CFU/mL, OD620 1 .3-1 .6). The culture was then diluted in PBS and then further diluted 1 : 1 in BHI broth containing hog gastric mucin to obtain the target inoculum density of 1 x 106 CFU per mL with 5% mucin. The actual colony counts were determined by plating dilutions to nutrient agar (NA) plates followed by 20 - 24 h incubation. The actual inoculum size was 1 .09 x 106 CFU/mL in BHI broth containing 5% mucin.
[0513] P. aeruginosa (ATCC 27853), Peritoneal Infection Model, LD90-100: P. aeruginosa (ATCC 27853) peritoneal Infection. Groups of 10 female ICR mice weighing 22 ± 2 g were used. Mice were inoculated intraperitoneally (IP) with P. aeruginosa (ATCC 27853) with an inoculum size at 5.4 x 105 CFU/mouse in 0.5 mL BHI broth containing 5% mucin. Test substance, GL-332 at 40 mg/kg, was administered intravenously (IV) once (QD) at 1 h after infection. GL-332, at 20 and 40 mg/kg, was administered IV twice (BID) with a 12 h interval (q12h) at 1 and 13 h after infection for a total dose of 40 or 80 mg/kg. The reference agent, gentamicin at 10 mg/kg, was administered subcutaneously (SC) BID q12h at 1 and 13 h after infection. Animals were observed for mortality twice daily (AM9:00/PM17:00) for 7 days after infection. Fisher’s exact test was used to assess the statistical significance in the survival of the treated animals compared to the vehicle control group at day 7 time point. The p value < 0.05 indicated a significant increase in the survival rates of the treated animals.
Results
[0514] The infection with P. aeruginosa ATCC 27853 at 5.4 c 105 CFU/mouse resulted in 100% mortality during the 7-day observation period. The IV administrations of GL-332, at 40 mg/kg QD, and at 20 and 40 mg/kg BID, resulted in significant increases in the survival relative to the vehicle control group (100% survival, p < 0.05) showing a protective effect. A significant increase in the survival rate (100% survival, p < 0.05) was also found in the SC dosing group of gentamicin at 10 mg/kg BID. ( See Table 10 and FIG. 4). As shown in Table 10 and FIG. 4, the survival rate for the mice administered the vehicle control rapidly decreased at day 1 ; however, the survival rate for GL-332 (QD and BID) was 100% after day 7, comparable to gentamicin. No overt adverse events were observed for 30 min after GL-332 at 20 and 40 mg/kg IV administrations in this study.
Table 10. P. aeruginosa ATCC 27853, peritoneal infection model, survival rate for 7 days
Figure imgf000206_0001
Animals were observed for mortality twice daily (AM9:00/PM17:00) for 7 days post infection.
*: Indicated a significant increase (p< 0.05) in the survival rates of the treated animals compared to the vehicle control group on Day 7 as determined by Fisher’s exact test
Conclusion
[0515] The results from this study show that there is statistically significant survival rate 7 days after infection of P. aeruginosa with IV administrations of GL-332, at each of the tested doses: 40 mg/kg QD, 20 mg/kg BID, and 40 mg/kg BID. This data indicates that GL-332 has a profound protective effect against bacterial infections.
Example 15: Stability of GL-332 to Enzymes Conferring Antibiotic Resistance
[0516] A major contributor to antibiotic resistance is b-lactamases enzymes that enable the inactivation of antimicrobial agents. A wide range of b-lactamases enzymes have been identified including the serine b-lactamases: CTX-M15 (Ambler class A), NDM-1 and IMP-1 (Ambler class B), KPC-2 and AmpC (Ambler class C), OXA-48 and OXA-24 (Ambler class D). Since each of the Ambler class enzymes A-C are found to be drug resistant, treatment options are limited and there is a need to develop antimicrobial agents that have activity against the enzymes.
[0517] The goal of this study was to evaluate GL-332 for its activity against each of the Ambler class A-D enzymes. The data show that GL332 is not degraded by OXA- 48 or OXA-24 (Ambler class D) and only slowly degraded by CTX-M-15 (Ambler class A) and KPC-2 (Amber class A), suggesting that GL-332 is not inactivated by clinically important extended spectrum b-lactamase and carbapenemase serine enzymes.
Methods
[0518] Stability of GL-332 to metallo^-lactamase (Ambler Class B). To determine the stability of GL-332 in the presence of the metallo^-lactamase, a solution having a concentration 75 mM GL332 was incubated with a solution comprising NDM-1 , IMP-1 or VIM-1 (enzyme concentration range 0.1 -2 pM). No hydrolysis of GL-332 by three enzymes occurred, suggesting that GL-332 is stable in the presence of the metallo-b- lactamase Amber class enzymes.
[0519] Stability of GL-332 Serine-Active Site b-lactamases. To determine the stability of GL-332 to serine-active site b-lactamases, a solution having a concentration 75 pM GL332 was incubated with a solution comprising KPC-2, OXA-24, OXA-48, CTX- M-15, and AmpC (enzyme concentration range 0.1 -2 pM). GL-332 was stable to OXA- 24 OXA-48 b-lactamases; however, GL-332 was completely hydrolyzed in the presence of CTX-M-15, allowing the determination of the molar absorptivity (ASM) of 4,300 M 1cnr 1 and then, further determination of the kinetic parameters of GL-332 in the presence of each of the enzymes.
[0520] Determination of the Kinetic Parameters. To evaluate GL-332’s stability from degradation by clinically important b-lactamases, the kinetic parameters were determined using the Hanes Plot Equation:
Figure imgf000207_0001
S is substrate concentration; Km is the partition ratio of the GL-332-enzyme Michaelis complex and Vmax = Kcat[ET], where Kcat is the catalytic turnover number and [ET] is the total enzyme concentration.
[0521] From these kinetic constants the enzyme’s catalytic efficiency can be assess by kCat/Km. Good substrates have kCat/Km values approaching that of diffusion, 108-109 M 1 sec1 and poor substrates have values many orders of magnitude less.
[0522] The experiments were collected in a 25 mM sodium phosphate buffer at pH of 7.0 (wavelength = 290 nm; sM=18,258M 1cnr1 at 290 nm; ASM = 4,300 M-1cnr1 at 290 nm).
[0523] Inhibition Model (determination of I C50 and K,- values). Competitive assays were next performed and the IC50 and equilibrium inhibition constant (Ki) values determined for GL-332 in the presence of OXA-48, OXA-24, CTZ-M-15, KPC-2, AmpC, NDM-1 , and IMP-1.
[0524] For class A, C, and D b-lactamases the assays were performed in a 25 mM sodium phosphate buffer at pH of 7.0 whereas the class B b-lactamases were performed in 20 mM HEPES and 20 mM ZnCL at a pH = 7.0. The concentrations for each of the b-lactamases used in the assays is provided in below in Table 11.
Table 11. 3-lactamases Concentrations for the Inhibition Assays
Figure imgf000208_0002
[0525] The data was then fit to the following equation:
Figure imgf000208_0001
Vi and Vo represent the initial rates of hydrolysis of Nitrocefin with or without inhibitor, respectively; I is the concentration of GL-332 acting as inhibitor or poor substrate; Ki is the inhibition constant; Km value is the Henri-Michael constant; S is the concentration of reporter substrate.
[0526] Nitrocefin is the chromogenic cephalosporin substrate that was used to monitor enzyme activity and has a max of 386 nm that shifts to 482 nm upon hydrolysis giving a striking visual color change from yellow to red.
Results
[0527] Table 12 summarizes the kinetic parameters determined for GL-332 in the presence of the serine-active site b-lactamases for CTZ-M-15, KPC-2, and AmpC as determined by the Hane’s plot equation. GL-332 is quite stable against AmpC catalyzed degradation with a turnover number of 4.5 xI O V1 and a very low catalytic efficiency of 6.1 x 101 M V1. While the poor activity of AmpC prevented measurement of Km directly, it was estimated from its Ki value. The modest Ki (Km) value of 74 mM suggests fair affinity for the enzyme’s active site, but very poor forward rate of catalysis (4.5 x103s- 1), consistent with the conclusion that GL-332 is a competitive inhibitor of the AmpC enzyme.
[0528] The serine carbapenemase, KPC-2 has a poor turnover number of 2.0 s 1 as does the extended spectrum b-lactamase CTX-M-15. The Km for KPC-2, 600 pM, suggests that the enzyme will be well below half saturation at physiological dosed concentrations, resulting in a poor enzyme efficiency of 3.3 x 103 M-1 s-1. Likewise, the Km value of CTX-M-15, 80 pM, suggests the enzyme would only be about half saturated at expected physiological concentrations yielding a modest catalytic efficiency of 3.5 x 104. These results are consistent with GL-332 being sufficiently stable against these enzymes to be clinically useful antibiotics.
Table 12. Summary of the kinetic parameters of GL-332 in the presence of CTZ-M-15, KPC-2, and AmpC
Figure imgf000209_0001
Figure imgf000210_0001
[0529] FIGS. 5A-5G linear fits to a model of competitive inhibition of GL-332 and AmpC, KPC-2, CTX-M-15, NDM-1 , IMP-1 , OXA-24, and OXA-48, respectively. Importantly, FIGs. 5A-5G reveal that GL-332 inhibited all of the b-lactamases via a competitive mechanism, indicating that these b-lactamases will not inhibit the antimicrobial activity of GL-332.
[0530] The analysis also showed favorable Ki and ICso values as enumerated below in Table 13. As shown in Table 13, GL-332 was able to most efficiently inhibited the class B b-lactamases, NDM-1 and IMP-1 as evidenced by the low Ki values of 90±5 mM and 26±2 mM, respectively.
Table 13. Ki and ICso by Ambler Classification
Figure imgf000210_0002
Conclusion
[0531] Overall, this study showed that GL322 is stable to Class B (NDM-1 and IMP-1 ) and Class D (OXA-24 and OXA-48) b-lactamases. However, the study showed that GL322 behaved as poor substrate of AmpC enzyme, which was able to hydrolyze the compound with poor catalytic efficiency (kCat/Km= 6.1 101 M-1S 1). GL-332 was also able to inhibit all b-lactamases via a competition mechanism (See FIGs. 5A-5G). It is also noteworthy that GL-322 was able to most efficiently inhibit Class B b-lactamases (lowest Ki values). And lastly, GL-322 inhibited all serine-active site enzymes with similar efficiency. [0532] Accordingly, the kinetic studies together with ICso values indicate that GL- 332 is a suitable drug candidate for the development of more potent b-lactamase inhibitors and is not susceptible to the inactivation by serine b-lactamases from the Ambler class A-D.
Example 16: Pharmacokinetics and Stability of GL337
[0533] In this study, the pharmacokinetics of subcutaneous (SC) and intravenous (IV) administration of GL-337 was evaluated in mice.
Methods
[0534] Sample Preparation for GL-337. The plasma samples were prepared by first adding a 10 pL of the GL-337 to 100 pL of the internal standard (100 ng/mL Labetalol & 100 ng/mL dexamethasone & 100 ng/mL tolbutamide & 100 ng/mL Verapamil & 100 ng/mL Glyburide & 100 ng/mL Celecoxib in CAN). The mixture was then vortexed and centrifuged at 12,000 rpm for 15 min at 4°C. A 30 pL of the supernatant was then mixed with 30 pL of water, vortexed, and then centrifuged at 4°C. A 8 pL of the sample was then injected for LC-MS/MS analysis.
[0535] Dilution Procedure. To dilute each sample by a factor of 40, an aliquot of 1 pL of sample was added to 39 pL of blank plasma.
[0536] LC-MS/MS Analysis. The data were collected by LC-MS/MS using ACQUITY UPLC HSS T3 1.8 pm 2.1 c 50 mm column at temperature of 60°C and flow rate of 0.65 mL/min (positive, ESI). GL-332 was observed at a [M+H]+ m/z of 354.6/172.3.
[0537] Study Details. The species used in this study were fasted, female CD-1 mice. The mice were injected SC and IV at a nominal dose of 20 mg/kg (administered 20.6 mg/kg) with GL-337. Both the IV and SC formulation had a 5 mg/mL concentration of GL-337 in pH 7 100 mM sodium bicarbonate buffer, clear solution. The experiments included 6 different mice denoted by M01 -M06: M01 -M03 for the IV injections and M04- M06 for the SC injections.
Results
[0538] The IV and SC pharmacokinetic parameters are enumerated in Tables 14 and 15, respectively. FIGs. 6A and 6B show the plasma concentration of GL-337 after IV and SC dosing at 20 mg/kg, respectively. And lastly, FIG. 6C shows the mean plasma concentration of GL-337 after IV and SC dosing. As demonstrated by FIGs. 6A-6C, GL-337 was rapidly absorbed at 20 mg/kg in the mice.
Table 14. IV Bioavailabilitv of GL-337 in Mouse in (nq/mU
Figure imgf000212_0001
Table 15. SC Bioavailabilitv of GL-337 in Mouse in (nq/mU
Figure imgf000212_0002
Figure imgf000213_0001
[0539] GL-337 was also assayed for its stability in plasma and was found to exhibit a high degree of stability. In a K2EDTA buffer, the half-life of GL-337 was greater than 240 min with 106% of GL-337 remaining at 120 min (T120). As a point of comparison, propantheline had only have half-life of 42.2 min with 13.6% of propantheline remaining at T120.
Conclusion
[0540] Overall, GL-337 exhibits bioavailability of SC and IV administered doses at 20 mg/kg. GL-337 further exhibits long half-lives, which suggests that GL-337 can be a candidate for treating bacterial infections at once daily dosing regimens.
[0541] The references cited above with respect to beta-lactamases are incorporated by reference herein for descriptions of various beta-lactamases and provide beta-lactamases in addition to those described herein which can be employed by one of ordinary skill in the art to further characterize the activity of compounds of Formula I against bacteria producing such beta-lactamases.
Figure imgf000214_0001
*Azt = aztreonam, Cef = ceftazidime, AC a combination of ceftazidime and the non-beta-lactam beta-lactamase inhibitor avibactam, MICs for AC measured by varying the amount of ceftazidime keeping the amount of avibactam constant at 4 mg/L.
Figure imgf000215_0001
*Azt = aztreonam
Figure imgf000216_0001
*Azt = aztreonam, Cef = ceftazidime, AC= a combination of ceftazidime and avibactam, MICs for AC measured by varying the amount of ceftazidime keeping the amount of avibactam constant at 4 mg/L.
Figure imgf000217_0001
*Azt = aztreonam.
Figure imgf000218_0001
Figure imgf000219_0001
Figure imgf000220_0001
# MIC50 is the antibiotic concentration that kills 50% of the panel of n bacteria; MIC90 is the antibiotic concentration that kills 90% of the panel of n bacteria. MIC range lists the lowest and highest MIC for the given panel.
*AC = a combination of ceftazidime and the non-beta-lactam beta-lactamase inhibitor avibactam, MICs for AC measured by varying the amount of ceftazidime keeping the amount of avibactam constant at 4 mg/L.
% Enterobacteriaceae include E. coli and K. pneumonia strains. REFERENCES
[1 ] Magiorakos, a; Srinivasan, A.; Carey, R. B.; Carmeli, Y.; Falagas, M. E.; Giske, C. G.; Harbarth, S.; Hindler, J. F. Bacteria : An International Expert Proposal for Interim Standard Definitions for Acquired Resistance. Microbiology 201 1 , 18 (3), 268-281.
[2] Spellberg, B.; Guidos, R.; Gilbert, D.; Bradley, J.; Boucher, H. W.; Scheld, W. M.; Bartlett, J. G.; Edwards, J. The Epidemic of Antibiotic-Resistant Infections: A Call to Action for the Medical Community from the Infectious Diseases Society of America. Clin. Infect. Dis. 2008, 46 (2), 155-164.
[3] Craig, W. A. State-of-the-Art Clinical Article: Pharmacokinetic/Pharmacodynamic Parameters: Rationale for Antibacterial Dosing of Mice and Men. Clin. Infect. Dis. 1998, 26 (1 ), 1-10.
[4] Montgomery, a B. The Antibiotic Pipeline. N. Engl. J. Med. 2004, 351 (19), 2019- 2020.
[5] Bakker-Woudenberg, I. a J. M.; ten Kate, M. T; Goessens, W. H. F.; Mouton, J. W. Effect of Treatment Duration on Pharmacokinetic/pharmacodynamic Indices Correlating with Therapeutic Efficacy of Ceftazidime in Experimental Klebsiella Pneumoniae Lung Infection. Antimicrob. Agents Chemother. 2006, 50 (9), 2919-2925.
[6] Boucher, H. W.; Talbot, G. H.; Bradley, J. S.; Edwards, J. E.; Gilbert, D.; Rice, L. B.; Scheld, M.; Spellberg, B.; Bartlett, J. Bad Bugs, No Drugs: No ESKAPE! An Update from the Infectious Diseases Society of America. Clin. Infect. Dis. 2009, 48 (1 ), 1-12.
[7] World Health Organization. Antimicrobial Resistance Global Report on Surveillance; Geneva, 2014.
[8] Liu, Y. Y.; Wang, Y.; Walsh, T. R.; Yi, L. X.; Zhang, R.; Spencer, J.; Doi, Y.; Tian, G.; Dong, B.; Huang, X.; et al. Emergence of Plasmid-Mediated Colistin Resistance Mechanism MCR-1 in Animals and Human Beings in China: A Microbiological and Molecular Biological Study. Lancet Infect. Dis. 2015, 16 (2), 161-168.
[9] Neu, H. C. The Crisis in Antibiotic Resistance. Science 1992, 257 (5073), 1064- 1073.
[10] Johnson, J. W.; Gretes, M.; Goodfellow, V. J.; Marrone, L.; Heynen, M. L.; Strynadka, N. C. J.; Dmitrienko, G. I. Cyclobutanone Analogues of Beta-Lactams Revisited: Insights into Conformational Requirements for Inhibition of Serine- and Metallo-Beta-Lactamases. J. Am. Chem. Soc. 2010, 132 (8), 2558-2560.
[11 ] Tioni, M. F.; Llarrull, L. I.; Poeylaut-Palena, A. a; Marti, M. a; Saggu, M.; Periyannan, G. R.; Mata, E. G.; Bennett, B.; Murgida, D. H.; Vila, A. J. Trapping and Characterization of a Reaction Intermediate in Carbapenem Hydrolysis by B. Cereus Metallo-Beta-Lactamase. J. Am. Chem. Soc. 2008, 130 (47), 15852-15863.
[12] Yuan, Q.; He, L.; Ke, H. A Potential Substrate Binding Conformation of beta- Lactams and Insight into the Broad Spectrum of NDM-1 Activity. Antimicrob. Agents Chemother. 2012, 56 (10), 5157-5163.
[13] E. P. Abraham, "A Glimpse of the Early History of the Cephalosporins," Reviews of Infectious Diseases, Vol. 1 , No. 1. (Jan. - Feb., 1979), pp. 99-105.
[14] E.P. Abraham, "Cephalosporins 1945-1986," Drugs 34 (Suppl. 2): 1 -14 (1987).
[15] Long, T. E.; Williams, J. T. Cephalosporins Currently in Early Clinical Trials for the Treatment of Bacterial Infections. Expert Opin. Investig. Drugs 2014, 23 (10), 1375- 1387.
[16] Zaffiri, L.; Gardner, J.; Toledo-Pereyra, L. H. History of Antibiotics. From Salvarsan to Cephalosporins. J. Investig. Surg. 2012, 25, 67-77.
[17] Peterson, L. R. Bad Bugs, No Drugs: No ESCAPE Revisited. Clin. Infect. Dis. 2009, 49, 992-993;
[18] IDSA. The 10 X '20 Initiative: Pursuing a Global Commitment to Develop 10 New Antibacterial Drugs by 2020. Clin. Infect. Dis. 2010, 50 (8), 1081-1083;
[19] Boucher, H. W.; Talbot, G. H.; Benjamin, D. K.; Bradley, J.; Guidos, R. J.; Jones, R. N.; Murray, B. E.; Bonomo, R. A.; Gilbert, D. 10 X '20 Progress-Development of New Drugs Active Against Gram-Negative Bacilli: An Update From the Infectious Diseases Society of America. Clin. Infect. Dis. 2013, 1-10;
[20] Pendleton, J. N.; Gorman, S. P.; Gilmore, B. F. Clinical Relevance of the ESKAPE Pathogens. Expert Rev. Anti. Infect. Ther. 2013, 11 (3), 297-308;
[21 ] WHO I WHO Publishes List of Bacteria for Which New Antibiotics Are Urgently Needed. WHO 2017; [22] Biggest Threats | Antibiotic/Antimicrobial Resistance | CDC see website www.cdc.gov/drugresistance/biggest_threats.html (accessed Jul 23, 2017);
[23] NIAID Emerging Infectious Diseases/Pathogens | NIH: National Institute of Allergy and Infectious Diseases see website www.niaid.nih.gov/research/emerging- infectious-diseases-pathogens.
[24] G.A. Jacoby (2009) AmpC Beta-Lacatamses Clin., Microbiol Rev. 22(1 ) 161 -182.
[25] Kopka et al. (2006) J. Med. Chem. 49(23):6704-6715
[26] Bush K and Jacoby GA (2010) Antimicro Agents Chemother, Vol. 54: p. 969- 976.
[27] Shaikh S. et al. (2015) Saudi Journal Biol, Sciences 22:90-101.
[28] Urban C. et al. (1994) Antimicrob. Agent. Chemother. 38:392-395.
[29] Chaves J. et al. (2001 ) Antimicrob. Agent. Chemother. 45:2856-2861.
[30] Hennequin C. et al. (Oct. 2011 ) Antimicrob. Agent. Chemother. 56:288-294.
[31 ] Aghamiri S. et al. (2014) ISRN Microbiology Article IS 941507 6 pages (available from web site: dx.doi.org/10.1155/2014/941507).
[32] Canton R. et al. (2012) Frontiers Microbiology, Vol. 3, article 110: 1 -1.
[33] Papp-Wallace K. et al (2015) Antimicrob. Agent. Chemother. 59:3710-3717.
[34] Alba, J. et al. (2005) Antimicrob. Agent. Chemother. 49:4760-4762.
[35] Raghunath d. (Nov. 2010) IJMR (Indian J. Med. Res.) 132(5):478-481.
[36] Saini A and Barsal, R. (2012) Advances Biol Chem 2:323-334.
[37] Poirel L. et al. (2000) Antimicrob. Agent. Chemother. April:891 -897.
[38] CLSI document M26-A. CLSI, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 1999.

Claims

I/We claim:
1 . A compound having the following chemical formula:
Figure imgf000224_0001
GL332 (482),
or a salt or solvate thereof.
2. The compound of claim 1 , wherein the compound is in one of two tautomeric forms.
3. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier.
4. The compound of claim 1 for use in the treatment or prevention of a microbial infection in a subject in need thereof.
5. A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the compound of claim 1 or the pharmaceutical composition of claim 3.
6. A compound of Formula I:
Figure imgf000224_0002
Formula I
or a salt or solvate thereof, where:
R is an acylamino group (R1CO-NH-) or an alkyl group optionally substituted with one or more groups selected from a hydroxy group, a protected hydroxyl group or a halogen;
M is a divalent cephem, penem or monobactam ring selected from:
Figure imgf000225_0001
where:
each RA is independently hydrogen, a C1 -C3 alkyl group or a C1 -C3 alkoxy group;
RB is hydrogen, a C1-C3 alkyl group or a C1 -C3-alkoxy group;
Rc is hydrogen, unsubstituted or substituted alkyl, unsubstituted or substituted aryl, alkoxy-substituted alkyl, acylalkoxy-substituted alkyl, acyloxy- substituted alkyl, a carboxyl protecting group or, when the CO2 group to which Rc is attached is negatively charged, a pharmaceutically acceptable cation;
Zi and Z2 are independently, S, SO, SO2, O, or C(RE)2, where each RE is independently hydrogen or a C1-C3 alkyl group;
L is an optional linker which is a substituted or unsubstituted ethylene or a substituted or unsubstituted phenyl group and n is 0, if the linker is absent, or is an integer ranging from 1 to 5, if the linker is present; the B ring is an unsubstituted or substituted phenyl ring or an unsubstituted or substituted 6-member heteroaryl ring having one to four ring nitrogens; and
R5, R6, R7, Re and Rg are independently selected from hydrogen, halogen, cyano, nitro, C1-C3 alkyl, C1 -C3 haloalkyl, amino, C1-C3 alkylamino, C1-C3 dialkylamino, and -CH2-X, with the exception that when the ring atom to which one of R5-R9 is bonded is a nitrogen, that R5-R9 is not present or is independently selected from hydrogen or C1 -C3 alkyl and at least one of R5 to Rg is -CH2-X, where X is an organic or inorganic leaving group.
7. The compound of claim 6, wherein M is Mi.
8. The compound of claim 7, wherein Zi is S or C(RE)2.
9. The compound of claim 6, wherein M is M2.
10. The compound of claim 9, wherein Z2 is S or C(RE)2.
1 1. The compound of claim 6, wherein M is M3.
12. The compound of claim 1 1 , wherein n is 0.
13. The compound of claim 1 1 , wherein n is 1 and L is an optionally substituted ethylene.
14. The compound of claim 13, wherein the M group and the B ring are substituted on the ethylene in the cis conformation.
15. The compound of claim 13, wherein the M group and the B ring are substituted on the ethylene in the trans conformation.
16. The compound of any one of claims 6-15, wherein R is an acylamino group.
17. The compound of any one of claims 6-15, wherein -CH2-X is:
Figure imgf000227_0001
wherein:
A+ is a leaving group containing a positively charged nitrogen;
Ri i , if present, is selected from hydrogen, hydroxyl, halogen, C1-C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1-C3 alkyl, unsubstituted or substituted C1-C3 alkoxy, cyano, and nitro groups or
R11 with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; and
D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two orthree heteroatoms (preferably N, S or O), where m is 1 or 0 to indicate presence or absence of linker D, where optional substitution for the D group is one or more halogen, oxo group, C1-C3 alkyl group, or C1-C3 alkoxy group.
18. The compound of any one of claims 6-15, wherein R is an acylamino group and -CH2-X is:
Figure imgf000228_0001
wherein:
A+ is a leaving group containing a positively charged nitrogen;
Ri i , if present, is selected from hydrogen, hydroxyl, halogen, C1-C3 alkyl, cyano, and nitro groups or Rn together with one of R12-R15 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic;
R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1-C3 alkyl, unsubstituted or substituted C1-C3 alkoxy, cyano, and nitro groups or
R11 with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached can form an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; and
D is an optional substituted divalent linker moiety containing 1 -6 carbon atoms and optionally one, two orthree heteroatoms (preferably N, S or O), where m is 1 or 0 to indicate presence or absence of linker D, where optional substitution for the D group is one or more halogen, oxo group, C1 -C3 alkyl group, or C1-C3 alkoxy group.
19. The compound of any one of claims 6-15, wherein -CH2-X is:
Figure imgf000228_0002
wherein: Rii , if present, is selected from hydrogen, hydroxyl, halogen, C1-C3 alkyl, cyano, and nitro groups;
R12-R15 are independently selected from hydrogen, hydroxyl, halide, carboxylate or ester thereof, acyl, unsubstituted or substituted aryl, unsubstituted or substituted arylalkyl, unsubstituted or substituted C1-C3 alkyl, unsubstituted or substituted C1-C3 alkoxy, cyano, and nitro groups; or
R11 with any one of R12-R15 or any two of R12-R15 together with the carbons to which they are attached forms an optionally substituted 5- or 6-member carbocyclic, or heterocyclic ring which may be saturated, partially unsaturated or aromatic; or
R12 and Ri3 or R13 and R14 or R14 and Ri 5 together with the carbons to which they are attached form an optionally substituted 5- or 6- member carbocyclic, or heterocyclic ring, which may be saturated, partially unsaturated or aromatic and optionally contains one or two additional heteroatoms;
R16 is selected from hydrogen and unsubstituted or substituted C1-C6 alkyl;
Ri7 is selected from hydrogen, and unsubstituted or substituted C1-C6 alkyl;
R18 is a divalent -(CH2) - moiety, where p is 1-6, wherein one or two CH2 groups are optionally replaced with one or more -0-, -S-, -CO-, -N(RN)CO-, or -CON(RN)-, where RN is hydrogen or a C1-C3 alkyl;
or Ri7 and Ris together with the nitrogen to which they are attached form a 5- or 6-member carbocylic or heterocyclic ring as shown by the dotted line, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms; or R18 and R15 together with the carbons to which they are attached form a 5- or 6-member carbocylic or heterocyclic ring, which ring is partially or fully unsaturated, unsubstituted or substituted, and optionally contains one or two additional heteroatoms;
where dotted lines indicate optional bonds.
20. A compound selected from the group consisting of Compounds 482, 487, 502, 503, 506, 507, 513, 481 , 478, 479, 490, 491 , 495, 500, 491-cis, and 495-cis.
21. A compound selected from the group consisting of Compounds 484, 489, 476, and 477.
22. A compound selected from the group consisting of Compounds 505 and
512.
23. A compound having the following chemical formula:
Figure imgf000230_0001
GL337 (487),
or a salt or solvate thereof.
24. A compound having the following chemical formula:
Figure imgf000230_0002
GL357,
or a salt or solvate thereof.
25. A compound having the following chemical formula:
Figure imgf000231_0001
GL353 (502),
or a salt or solvate thereof.
26. The compound of any one of claims 6 to 25, wherein the compound is in one of two tautomeric forms.
27. A pharmaceutical composition comprising a compound of any one of claims 6-26 and a pharmaceutically acceptable carrier.
28. A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 6-26 or a pharmaceutical composition of claim 27.
29. Use of a compound of any one of claims 6-26 or a pharmaceutical composition of claim 26 for treating or preventing a bacterial infection.
30. Use of a compound of any one of claims 6-26 for making a medicament for treating or preventing a bacterial infection.
31 . A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the compound:
Figure imgf000231_0002
GL337 (487), or a salt or solvate thereof.
32. A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the compound:
Figure imgf000232_0002
or a salt or solvate thereof.
33. A method for treating or preventing a bacterial infection in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the compound:
Figure imgf000232_0001
GL357,
or a salt or solvate thereof.
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Publication number Priority date Publication date Assignee Title
CN113045573A (en) * 2021-03-09 2021-06-29 南开大学 Probe compound resistant to carbapenem antibiotic germs and application

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4782155A (en) * 1986-03-19 1988-11-01 Banyu Pharmaceutical Co., Ltd. Cephalosporin derivatives, processes for their preparation and antibacterial agents
US5126336A (en) * 1990-08-23 1992-06-30 Bristol-Myers Squibb Company Antibiotic c-3 catechol-substituted cephalosporin compounds, compositions and method of use thereof
WO2010083725A1 (en) * 2009-01-20 2010-07-29 山东轩竹医药科技有限公司 Cephalosporin derivatives containing substituted nitrogen-containing fused heterocyclic ring
US20130102583A1 (en) * 2010-04-05 2013-04-25 Shionogi & Co., Ltd. Cephem compound having catechol group
US20190100534A1 (en) * 2017-10-04 2019-04-04 Gladius Pharmaceuticals, Inc. Cephem Compounds with Latent Reactive Groups

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4782155A (en) * 1986-03-19 1988-11-01 Banyu Pharmaceutical Co., Ltd. Cephalosporin derivatives, processes for their preparation and antibacterial agents
US5126336A (en) * 1990-08-23 1992-06-30 Bristol-Myers Squibb Company Antibiotic c-3 catechol-substituted cephalosporin compounds, compositions and method of use thereof
WO2010083725A1 (en) * 2009-01-20 2010-07-29 山东轩竹医药科技有限公司 Cephalosporin derivatives containing substituted nitrogen-containing fused heterocyclic ring
US20130102583A1 (en) * 2010-04-05 2013-04-25 Shionogi & Co., Ltd. Cephem compound having catechol group
US20190100534A1 (en) * 2017-10-04 2019-04-04 Gladius Pharmaceuticals, Inc. Cephem Compounds with Latent Reactive Groups

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE Pubchem 4 December 2011 (2011-12-04), ANONYMOUS: "(6S)-7-[[2-(2-Amino-1,3-thiazol-4-yl)-2-(2-carboxypropan-2-yloxyimino)acetyl]amino]-3-(2,6-dibromo-3,4-dihydroxyphenyl)-7-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid", XP055745419, retrieved from NCBI Database accession no. 53804255 *
FRANK DININNO ,DAVID A MUTHARD ,THOMAS N SALZMANN: "The synthesis and antibacterial activity of 2-para-quarternary ammoniomethylphenyl-carbapenems", BIORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 3, no. 11, 30 November 1993 (1993-11-30), pages 2187 - 2192, XP026630323, ISSN: 0960-894X, DOI: 10.1016/S0960-894X(01)80923-9 *
S NAKAGAWA, M SANADA, K MATSUDA, T HASHIZUME, Y ASAHI, R USHIJIMA, N OHTAKE, N TANAKA: "In vitro and in vivo antibacterial activities of BO-1341, a new antipseudomonal cephalosporin", ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, vol. 33, no. 9, 1 September 1989 (1989-09-01), pages 1423 - 1427, XP055745417, DOI: 10.1128/AAC.33.9.1423 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113045573A (en) * 2021-03-09 2021-06-29 南开大学 Probe compound resistant to carbapenem antibiotic germs and application

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