WO2017106552A1 - Potentiators of beta-lactam antibiotics and combination therapy - Google Patents

Potentiators of beta-lactam antibiotics and combination therapy Download PDF

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WO2017106552A1
WO2017106552A1 PCT/US2016/067015 US2016067015W WO2017106552A1 WO 2017106552 A1 WO2017106552 A1 WO 2017106552A1 US 2016067015 W US2016067015 W US 2016067015W WO 2017106552 A1 WO2017106552 A1 WO 2017106552A1
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compound
pyridinyl
ortho
ome
ocf3
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PCT/US2016/067015
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French (fr)
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Shahriar Mobashery
Enrico SPERI
Mayland Chang
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University Of Notre Dame Du Lac
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    • C07C57/52—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing halogen
    • C07C57/58—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing halogen containing six-membered aromatic rings
    • C07C57/60—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing halogen containing six-membered aromatic rings having unsaturation outside the rings
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
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    • A61K31/425—Thiazoles
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    • A61K31/545—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, cefaclor, or cephalexine
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Definitions

  • Staphylococcus aureus is a Gram-positive bacterium commonly found on the skin and in moist areas, such as the nasal cavity, yet it is often broadly resistant to many antibiotics, ⁇ - Lactam antibiotics were the drugs of choice for treatment of infection by S. aureus, but a variant of this organism, methicillin-resistant Staphylococcus aureus (MRS A) emerged in 1961, which exhibited resistance to the entire class of ⁇ -lactams. This organism has been a global clinical problem for over half a century. The molecular basis for the broad resistance of MRS A to ⁇ - lactams, which is incidentally inducible, was traced to a set of genes within the bla and mec operons.
  • the BlaRl (or the cognate MecRl) protein is a ⁇ -lactam antibiotic sensor/signal transducer, which communicates the presence of the antibiotic in the milieu to the cytoplasm in a process that is largely not understood (Staude et al., Biochemistry 2015, 54, 1600-1610).
  • MRSA methicillin-resistant Staphylococcus aureus
  • an antibiotic sensor/signal transducer is phosphorylated on exposure to ⁇ -lactam antibiotics. This event is critical for the onset of the biochemical events that unleash induction of antibiotic resistance.
  • the phosphorylation and the antibiotic-resistance phenotype are abrogated in the presence of novel inhibitors that restore susceptibility of the organism to ⁇ -lactam antibiotics.
  • the invention provides compounds, compositions, and methods for reducing, preventing, overcoming, and/or abrogating resistance to ⁇ -lactam antibiotics, and methods of treating bacterial infections caused by antibiotic resistant bacteria, particularly bacteria that can develop resistance to ⁇ -lactam antibiotics. Accordingly, the invention provides compounds that potentiate the antibacterial activity of ⁇ -lactam antibiotics toward ⁇ -lactam-resistant antibiotics.
  • the invention therefore provides compositions and methods for increasing the sensitivity of bacterial pathogens to antibiotics (e.g., potentiating the antibiotics), including ⁇ -lactam antibiotics.
  • the invention provides a method for increasing the sensitivity of bacterial pathogens to ⁇ -lactam antibiotics by contacting the bacterial pathogen with one or more compounds described herein.
  • the bacterial pathogen is MSRA.
  • the bacterial pathogen is Enterococcus faecalis or other bacterial pathogen described herein.
  • the invention also provides compositions and methods for increasing the susceptibility of Gram positive or Gram negative pathogens to ⁇ -lactam antibiotics.
  • Various embodiments provide pharmaceutical compositions, therapeutic formulations, product combination, or kits for use against MRSA infections comprising a compound described herein and one or more ⁇ - lactam antibiotics.
  • the compounds and methods can be used for inhibiting the growth of bacteria, for example, Staphylococcus aureus.
  • the Staphylococcus aureus is resistant to, or sensitive to, methicillin, other ⁇ -lactams, macrolides, lincosamides, aminoglycosides, or a combination thereof.
  • the invention further provides methods for increasing the sensitivity of Staphylococcus aureus to methicillin, other ⁇ -lactams, macrolides, lincosamides, or aminoglycosides.
  • the methods can include administering an effective amount of a compound, a pair of compounds, or composition described herein.
  • X is O, CH2, S, SO2, or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO2H, CC Me, CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or S02-(pyridinyl);
  • n 1, 2, 3, or 4;
  • n 1, 2, 3, 4, or 5;
  • each R 2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, CC Me, NO2, OCF 3 , or SCF3, or two R 2 groups form an ortho-fused methylenedioxy; and
  • each R 3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, C0 2 Me, NO2, OCF 3 , or SCF3, or two R 3 groups form an ortho-fused methylenedioxy;
  • X is O. In another embodiment, X is CH2. In yet another embodiment, X is S. In yet a further embodiment, X is a direct bond. In yet a further embodiment, X is SO2.
  • R 1 is CN or SO2CH3.
  • R 2 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R 2 groups form an ortho-fused
  • R 3 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R 2 groups form an ortho-fused methylenedioxy.
  • X is O, CH2, S, or a direct bond
  • R 1 is CN or SO2CH3
  • m is 1 or 2
  • n is 1 or 2.
  • R 2 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R 2 groups form an ortho-fused methylenedioxy
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R 3 groups form an ortho-fused
  • the compounds of Formula I include the various compounds of Formulas II and III, including their sub-Formulas II-A, II-B, II-C, II-D, III, III-A, and III-B. Accordingly, the invention provides a compound of Formula I wherein the compound is a compound of Formula II:
  • X is O, CH2, S. or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S0 2 -(pyridinyl); m is 1, 2, 3, or 4;
  • n 1, 2, 3, 4, or 5;
  • R 2 is H, F. OMe, OEt, OCF3, SCF3. N-morpholinyl, or pyridinyl. or two R 2 groups form an ortho-fused methylenedioxy;
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, N-mo holinyl, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention also provides compounds of Formula II wherein the compound is a compound of Formula II-A:
  • R 1 is CN. SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S0 2 -(pyridinyl); m is 1 or 2;
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R 2 groups form an ortho-fused methylenedioxy;
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention also provides compounds of Formula II wherein the compound is a compound of Formula II-B:
  • R 1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S0 2 -(pyridinyl); m is 1 or 2;
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R 2 groups form an ortho-fused methylenedioxy;
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention also provides compounds of Formula II wherein the compound is a compound of Formula II-C:
  • X is O, CH2, S, or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S0 2 -(pyridinyl); m is 1 or 2;
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R 2 groups form an ortho-fused methylenedioxy;
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention also provides compounds of Formula II wherein the compound is a compound of Formula II-D:
  • R 1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S0 2 -(pyridinyl); m is 1 or 2;
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R 2 groups form an ortho-fused methylenedioxy;
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention also provides compounds of Formula I wherein the compound is a compound of Formula III:
  • X is O, CH2, S, or a direct bond
  • R 1 is CN. SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S0 2 -(2- pyridinyl);
  • n 1 or 2;
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF3, SCF3, or N-morpholinyl
  • R 3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R 3 groups form an ortho-fused methylenedioxy;
  • the invention further provides compounds of Formula III wherein the compound is a compound of Formula III-A:
  • X is O, CH2, S, or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S0 2 -(2- pyridinyl);
  • n 1 or 2;
  • R 2 is H, F, OMe, OEt, OCF 3 , SCF 3 , or N-morpholinyl
  • X is O, CH2, S, or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO(aryl), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); n is 2;
  • R 2 is F
  • R 3 is CI
  • compositions comprising a compound of any one of Formulas I-III and/or Figure 3 in combination with a pharmaceutically acceptable diluent, excipient, or carrier.
  • compositions comprising a compound of any one of Formulas I-III and/or Figure 3 in combination with a ⁇ -lactam antibiotic.
  • ⁇ -lactam antibiotic is ceftadizim, ceftaroline, ceftazidime, meropenem, oxacillin, or penicillin, or another antibiotic recited herein.
  • the invention further provides a method to reverse the methicillin-resistant phenotype in bacteria comprising contacting methicillin-resistant Staphylococcus aureus (MRSA) with an effective amount of a compound described herein, thereby rendering MRSA susceptible to ⁇ - lactam antibiotics.
  • MRSA methicillin-resistant Staphylococcus aureus
  • the invention yet further provides a method to inhibit or kill methicillin-resistant
  • Staphylococcus aureus comprising contacting the MRSA with an amount of a compound described herein effective to reverse the methicillin-resistant phenotype, and contacting the MRSA with an effective antibacterial amount of a ⁇ -lactam antibiotic.
  • the invention provides a method to reduce or attenuate the minimum inhibitory concentration (MIC) of a ⁇ -lactam antibiotic comprising contacting a bacterium with an effective amount of a compound described herein in combination with contacting the bacterium with a ⁇ -lactam antibiotic.
  • the invention therefore provides for the use of a compound described herein for preparing a medicament to treat a bacterial infection.
  • the bacterial infection can be, for example, a methicillin-resistant Staphylococcus aureus (MRSA) infection.
  • MRSA methicillin-resistant Staphylococcus aureus
  • the administration of the compound described herein can be concurrent or sequential with an antibiotic, for example, a ⁇ -lactam antibiotic, including the specific ⁇ -lactam antibiotics described herein.
  • Further embodiments relate to methods of ameliorating and/or treating a bacterial infection that can include administering to a subject suffering from the bacterial infection an effective amount of one or more compounds of Formulas I-III, or a pharmaceutical composition that includes one or more compounds of Formulas I-III, or a pharmaceutically acceptable salt thereof.
  • Other embodiments described herein relate to using one or more compounds of Formulas I-III in the manufacture of a medicament for ameliorating and/or treating a bacterial infection.
  • Still other embodiments described herein relate to compounds of Formulas I-III that can be used for ameliorating and/or treating a bacterial infection.
  • bacterial infection can be an S. aureus infection, for example, a MRSA infection.
  • the invention thus provides novel compounds of Formulas I-III, intermediates for the synthesis of compounds of Formulas I-III, as well as methods of preparing compounds of Formulas I-III.
  • the invention also provides compounds of Formulas I-III that are useful as intermediates for the synthesis of other useful compounds.
  • the invention provides for the use of the compounds and compositions described herein in medical therapy.
  • the compounds of Formulas I-III can be used in the manufacture of medicaments useful for the treatment of bacterial infections in a mammal, such as a human.
  • Compositions and medicaments described herein can include a pharmaceutically acceptable diluent, excipient, or carrier.
  • Antimicrobial resistance is one of the most serious threats to global public health today.
  • ⁇ -Lactam antibiotics were the preferred antibiotics for treatment of infections by S. aureus, but emergence of Methicillin-resistant Staphylococcus aureus (MRS A) in 1961 made these drugs obsolete within a short time.
  • MRSA Methicillin-resistant Staphylococcus aureus
  • MRSA exhibits phosphorylation of certain proteins. Interference with phosphorylation or other critical pathways reverses the antibiotic-resistance phenotype.
  • MRSA methicillin-resistant Staphylococcus aureus
  • ⁇ -lactam antibiotics are phosphorylated on exposure to ⁇ -lactam antibiotics. This event is critical for the onset of the biochemical events that unleash induction of antibiotic resistance.
  • the phosphorylation and the antibiotic-resistance phenotype may be abrogated in the presence of novel inhibitors described herein, which inhibitors restore susceptibility of the organism to ⁇ - lactam antibiotics.
  • the invention thus provides compounds and methods for abrogating antibiotic resistance to ⁇ -lactam antibiotics, thereby potentiating the activity of a ⁇ -lactam antibiotic. Definitions
  • references in the specification to "one embodiment”, “an embodiment”, etc., indicate that the embodiment described may include a particular aspect, feature, structure, moiety, or characteristic, but not every embodiment necessarily includes that aspect, feature, structure, moiety, or characteristic. Moreover, such phrases may, but do not necessarily, refer to the same embodiment referred to in other portions of the specification. Further, when a particular aspect, feature, structure, moiety, or characteristic is described in connection with an embodiment, it is within the knowledge of one skilled in the art to affect or connect such aspect, feature, structure, moiety, or characteristic with other embodiments, whether or not explicitly described.
  • the term "and/or” means any one of the items, any combination of the items, or all of the items with which this term is associated.
  • the phrases "one or more” and “at least one” are readily understood by one of skill in the art, particularly when read in context of its usage. For example, the phrase can mean one, two, three, four, five, six, ten, 100, or any upper limit approximately 10, 100, or 1000 times higher than a recited lower limit.
  • one or more substituents on a phenyl ring refers to one to five, or one to four, for example if the phenyl ring is disubstituted.
  • the term “about” can refer to a variation of ⁇ 5%, ⁇ 10%, ⁇ 20%, or ⁇ 25% of the value specified. For example, “about 50" percent can in some embodiments carry a variation from 45 to 55 percent.
  • the term “about” can include one or two integers greater than and/or less than a recited integer at each end of the range. Unless indicated otherwise herein, the term “about” is intended to include values, e.g., weight percentages, proximate to the recited range that are equivalent in terms of the functionality of the individual ingredient, the composition, or the embodiment.
  • the term about can also modify the end-points of a recited range as discussed above in this paragraph.
  • ranges recited herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof, as well as the individual values making up the range, particularly integer values.
  • a recited range e.g., weight percentages or carbon groups
  • Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, or tenths. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc.
  • alkyl refers to a straight- or branched-chain alkyl group having from 1 to about 20 carbon atoms in the chain.
  • the alkyl group can be a (Ci-C2o)alkyl, a (Ci- Ci2)alkyl, (Ci-C8)alkyl, (Ci-C6)alkyl, or (Ci-C4)alkyl.
  • alkyl groups examples include methyl (Me), ethyl (Et), ⁇ -propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (t- u), pentyl, isopentyl, fert-pentyl, hexyl, isohexyl, and groups that in light of the ordinary skill in the art and the teachings provided herein would be considered equivalent to any one of the foregoing examples.
  • Alkyl groups can be optionally substituted or unsubstituted, and optionally partially unsaturated, such as in an alkenyl group.
  • cycloalkyl refers to cyclic alkyl groups of, for example, from 3 to 10 carbon atoms having a single cyclic ring or multiple condensed rings.
  • Cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantyl, pinenyl, and the like.
  • the cycloalkyl group can be monovalent or divalent, and can be optionally substituted, for example, by one or more alkyl groups.
  • the cycloalkyl group can optionally include one or more cites of unsaturation, for example, the cycloalkyl group can include one or more carbon-carbon double bonds, such as, for example, 1-cyclopent-l-enyl, l-cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, l-cyclohex-3-enyl, and the like.
  • alkoxy refers to the group alky 1-0-, where alkyl is as defined herein.
  • Preferred alkoxy groups include, e.g., methoxy, ethoxy, w-propoxy, wo-propoxy, w-butoxy, fert-butoxy, sec-butoxy, w-pentoxy, w-hexoxy, 1,2-dimethylbutoxy, and the like.
  • cycloalkyl refers to cyclic alkyl groups of, for example, from 3 to 10 carbon atoms having a single cyclic ring or multiple condensed rings.
  • Cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantyl, pinenyl, and the like.
  • the cycloalkyl group can be monovalent or divalent, and can be optionally substituted, for example, by one or more alkyl groups.
  • the cycloalkyl group can optionally include one or more cites of unsaturation, for example, the cycloalkyl group can include one or more carbon-carbon double bonds, such as, for example, 1-cyclopent-l-enyl, l-cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, l-cyclohex-3-enyl, and the like.
  • alkoxy refers to the group alky 1-0-, where alkyl is as defined herein.
  • Preferred alkoxy groups include, e.g., methoxy, ethoxy, w-propoxy, wo-propoxy, w-butoxy, fert-butoxy, sec-butoxy, w-pentoxy, w-hexoxy, 1,2-dimethylbutoxy, and the like.
  • heteroaryl refers to a monocyclic, bicyclic, or tricyclic ring system containing one, two, or three aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring.
  • the heteroaryl can be unsubstituted or substituted, for example, with one or more, and in particular one to three, substituents, as described in the definition of "substituted” .
  • Typical heteroaryl groups contain 2-20 carbon atoms in the ring skeleton in addition to the one or more heteroatoms.
  • heteroaryl groups include, but are not limited to, 2H-pyrrolyl, 3H-indolyl,
  • phenothiazinyl phenoxathiinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, thiadiazolyl, thianthrenyl, thiazolyl, thienyl, triazolyl, tetrazolyl, and xanthenyl.
  • heteroaryl denotes a monocyclic aromatic ring containing five or six ring atoms containing carbon and 1, 2, 3, or 4 heteroatoms independently selected from non-peroxide oxygen, sulfur, and N(Z) wherein Z is absent or is H, O, alkyl, aryl, or (Ci-C6)alkylaryl.
  • heteroaryl denotes an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benz-derivative or one derived by fusing a propylene, trimethylene, or tetramethylene diradical thereto.
  • the heteroaryl can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, acetamido, acetoxy, acetyl, benzamido, benzenesulfinyl, benzenesulfonamido, benzenesulfonyl, benzenesulfonylamino, benzoyl, benzoylamino, benzoyloxy, benzyl,
  • R x and R y are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl, or hydroxy.
  • the nitrogen of any indolyl ring can be N-substituted to provide an N-alkyl, N-methyl, or N-protecting group indolyl compound.
  • a heteroaryl can also be substituted with a substituent as described in the substituents definition below.
  • heterocycle is a monocyclic, bicyclic, or tricyclic group containing one or more heteroatoms selected from the group oxygen, nitrogen, and sulfur.
  • heterocycle groups include 1,3-dihydrobenzofuran, 1 ,3-dioxolane, 1 ,4-dioxane, 1 ,4-dithiane, 2H-pyran, 2- pyrazoline, 4H-pyran, chromanyl, imidazolidinyl, imidazolinyl, indolinyl, isochromanyl, isoindolinyl, morpholine, piperazinyl, piperidine, piperidyl, pyrazolidine, pyrazolidinyl, pyrazolinyl, pyrrolidine, pyrroline, quinuclidine, and thiomorpholine.
  • the heterocycle can optionally be a divalent radical, thereby providing a heterocyclene.
  • the heterocycle can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifiuoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, acetamido, acetoxy, acetyl, benzamido, benzenesulfinyl, benzenesulfonamido, benzenesulfonyl, benzenesulfonylamino, benzoyl, benzoylamino, benzoyloxy, benzyl,
  • R x and R y are independently ⁇ , alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl, or hydroxy.
  • a heterocycle can also be substituted with a substituent as described in the substituents definition below.
  • nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containing
  • Protecting Groups Compounds of the invention can further include one or more suitable protecting groups.
  • protecting group refers to any group that, when bound to an sp- center, a hydroxyl, nitrogen, or other heteroatom prevents undesired reactions from occurring at this group and that can be removed by conventional chemical or enzymatic steps to reestablish the 'unprotected' moiety, such as an alkyne, hydroxyl, nitrogen, or other heteroatom group.
  • the particular removable group employed is often interchangeable with other groups in various synthetic routes.
  • Certain removable protecting groups include conventional substituents such as, for example, allyl, benzyl, acetyl, chloroacetyl, thiobenzyl, benzylidine, phenacyl, methyl methoxy, silicon protecting groups ("silyl ethers") (e.g., trimethylsilyl (TMS), /-butyl- diphenylsilyl (TBDPS), triisopropylsilyl (TIPS), or /-butyldimethylsilyl (TBS)) and any other group that can be introduced chemically onto a hydroxyl or other moiety and later selectively removed either by chemical or enzymatic methods in mild conditions compatible with the nature of the product.
  • sil ethers e.g., trimethylsilyl (TMS), /-butyl- diphenylsilyl (TBDPS), triisopropylsilyl (TIPS), or /-butyldimethylsilyl (TBS)
  • halogen refers to chlorine, fluorine, bromine or iodine.
  • halo refers to chloro, fluoro, bromo or iodo.
  • each can further include one or more (e.g., 1, 2, 3, 4, 5, or 6) substituents. It is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
  • substituted means that a specified group or moiety can bear one or more (e.g., 1, 2, 3, 4, 5, or 6) substituents.
  • unsubstituted means that the specified group bears no substituents.
  • optionally substituted means that the specified group is unsubstituted or substituted by one or more substituents, and elements of the Formulas described herein can be optionally substituted. Where the term “substituted” is used to describe a structural system, the substitution is meant to occur at any valency-allowed position on the system.
  • suitable substituent groups include one or more of alkyl, alkenyl, alkynyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, aroyl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, alkylamino, dialkylamino, trifluoromethylthio, difluoromethyl, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, arylsulfinyl, arylsulfonyl, heteroarylsulfinyl, heteroarylsulfonyl, heterocyclesulfinyl
  • any one of the above groups can be included or excluded from a variable (e.g., groups R 2 and R 3 ) or from a group of substituents.
  • pharmaceutically acceptable salts refers to ionic compounds, wherein a parent non-ionic compound is modified by making acid or base salts thereof.
  • examples of pharmaceutically acceptable salts include, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • the pharmaceutically acceptable salts include conventional non-toxic salts and quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • Non-toxic salts can include those derived from inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, phosphoric, nitric and the like.
  • Salts prepared from organic acids can include those such as acetic, 2-acetoxybenzoic, ascorbic, behenic, benzenesulfonic, benzoic, citric, ethanesulfonic, ethane disulfonic, formic, fumaric, gentisinic, glucaronic, gluconic, glutamic, gly colic, hydroxymaleic, isethionic, isonicotinic, lactic, maleic, malic, mesylate or methanesulfonic, oxalic, pamoic (l, l '-methylene-bis-(2- hydroxy-3-naphthoate)), pantothenic, phenylacetic, propionic, salicylic, sulfanilic,
  • the pharmaceutically acceptable salts of the compounds described herein can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of many suitable salts are found in Remington: The Science and Practice of Pharmacy, 21 st edition, Lippincott, Williams & Wilkins, (2005).
  • solvate refers to a solid compound that has one or more solvent molecules associated with its solid structure. Solvates can form when a solid compound is crystallized from a solvent, wherein one or more solvent molecules become an integral part of the solid crystalline matrix.
  • the compounds of the formulas described herein can be solvates, for example, ethanol solvates.
  • Another type of a solvate is a hydrate.
  • a "hydrate” likewise refers to a solid compound that has one or more water molecules intimately associated with its solid or crystalline structure at the molecular level.
  • a hydrate is a specific type of a solvate. Hydrates can form when a compound is solidified or crystallized in water, wherein one or more water molecules become an integral part of the solid crystalline matrix.
  • the compounds of the formulas described herein can be hydrates.
  • diluent refers to a pharmacologically inert substance that is nevertheless suitable for human consumption that serves as an excipient in the inventive dosage form.
  • a diluent serves to dilute the API in the inventive dosage form, such that tablets of a typical size can be prepared incorporating a wide range of actual doses of the API.
  • excipient refers to an ingredient of the dosage form that is not medicinally active, but serves to dilute the API, assist in dispersion of the tablet in the patient's stomach, bind the tablet together, and serve other functions like stabilizing the API against decomposition.
  • contacting refers to the act of touching, making contact, or of bringing to immediate or close proximity, including at the cellular or molecular level, for example, to bring about a physiological reaction, a chemical reaction, or a physical change, e.g., in a solution, in a reaction mixture, in vitro, or in vivo (e.g., by administration to a patient).
  • an “effective amount” refers to an amount effective to treat a disease, disorder, and/or condition, or to bring about a recited effect.
  • an effective amount can be an amount effective to reduce the progression or severity of the condition or symptoms being treated. Determination of a therapeutically effective amount is well within the capacity of persons skilled in the art, especially in light of the detailed disclosure provided herein.
  • the term "effective amount” is intended to include an amount of a compound described herein, or an amount of a combination of compounds described herein, e.g., that is effective to treat or prevent a disease or disorder, or to treat the symptoms of the disease or disorder, in a host.
  • an “effective amount” generally means an amount that provides the desired effect.
  • the term "effective amount" can refer to an amount of compound or composition, which upon administration, is capable of reducing or preventing proliferation of a bacteria, reducing or preventing symptoms associated with a bacterial infection, reducing the likelihood of bacterial infection, or preventing bacterial infection.
  • the subject is treated with an amount of a therapeutic composition sufficient to reduce a symptom of a disease or disorder, such as an infection, by at least about 25%, about 50%, about 75%, or about 90%.
  • treating can include (i) preventing a disease, pathologic or medical condition from occurring (e.g., prophylaxis); (ii) inhibiting the disease, pathologic or medical condition or arresting its development; (iii) relieving the disease, pathologic or medical condition; and/or (iv) diminishing symptoms associated with the disease, pathologic or medical condition.
  • the terms “treat”, “treatment”, and “treating” can extend to prophylaxis and can include prevent, prevention, preventing, lowering, stopping or reversing the progression or severity of the condition or symptoms being treated.
  • treatment can include medical, therapeutic, and/or prophylactic administration, as appropriate.
  • compositions can be used to treat infections by drug-resistant strains of bacteria, for example MRSA (methicillin resistant S. aureus), MRSE (methicillin resistant S. epidermidis), PRSP (penicillin resistant S. pneumoniae), VIRSA (vancomycin intermittently resistant
  • MRSA methicillin resistant S. aureus
  • MRSE methicillin resistant S. epidermidis
  • PRSP penicillin resistant S. pneumoniae
  • VIRSA vancomycin intermittently resistant
  • the invention provides a method for killing or inhibiting growth of gram positive bacteria comprising contacting gram positive bacteria with a compound or composition described herein, thereby killing or inhibiting the growth of the bacteria.
  • the contacting can be performed in vivo in a human or animal, or in vitro, for example, in an assay.
  • the gram positive bacteria can be of the genus Enterococcus or Staphylococcus.
  • the bacteria is a drug-resistant strain of the genus Staphylococcus.
  • the bacteria is a methicillin-resistant
  • MRSA Staphylococcus aureus
  • the bacterial infection may be due to Gram-positive bacteria, including, but not limited to, methicillin resistant Staphylococcus aureus (MRSA), community- acquired methicillin resistant Staphylococcus aureus (CAMRSA), vancomycin-intermediate- susceptible Staphylococcus aureus (VISA), methicillin-resistant coagulase-negative bacteria.
  • MRSA methicillin resistant Staphylococcus aureus
  • CAMRSA community- acquired methicillin resistant Staphylococcus aureus
  • VSA vancomycin-intermediate- susceptible Staphylococcus aureus
  • methicillin-resistant coagulase-negative bacteria including, but not limited to, methicillin resistant Staphylococcus aureus (MRSA), community- acquired methicillin resistant Staphylococcus aureus (CAMRSA), vancomycin-intermediate- susceptible Staphylococcus aureus (VISA), methicillin-resistant coagulase-negative
  • MR-CoNS vancomycin-intermediate-susceptible coagulase-negative staphylococci
  • VI-CoNS vancomycin-intermediate-susceptible coagulase-negative staphylococci
  • MSSA methicillin susceptible Staphylococcus aureus
  • Streptococcus pneumoniae including penicillin-resistant strains [PRSP]) and multi-drug resistant strains [MDRSP]
  • Streptococcus agalactiae Streptococcus pyogenes and
  • the bacterial infection may include, but is not limited to, complicated skin and skin structure infections (cSSSI); community acquired pneumonia (CAP); complicated intra-abdominal infections, such as, complicated appendicitis, peritonitis, complicated cholecystitis and complicated diverticulitis; uncomplicated and complicated urinary tract infections, such as, pyelonephritis; and respiratory and other nosocomial infections.
  • cSSSI complicated skin and skin structure infections
  • CAP community acquired pneumonia
  • complicated intra-abdominal infections such as, complicated appendicitis, peritonitis, complicated cholecystitis and complicated diverticulitis
  • uncomplicated and complicated urinary tract infections such as, pyelonephritis
  • respiratory and other nosocomial infections such as, pyelonephritis.
  • infection refers to the invasion of the host by germs (e.g., bacteria) that reproduce and multiply, causing disease by local cell injury, release of poisons, or germ- antibody reaction in the cells.
  • germs e.g., bacteria
  • the compounds and compositions described herein can be used to treat a gram positive bacterial infection, for example, in a mammal, such as a human.
  • inhibitor refers to the slowing, halting, or reversing the growth or progression of a disease, infection, condition, or group of cells.
  • the inhibition can be greater than about 20%, 40%, 60%, 80%, 90%, 95%, or 99%, for example, compared to the growth or progression that occurs in the absence of the treatment or contacting.
  • Protein phosphorylation and its contribution to many regulatory events are widely known in eukaryotes. However, the same information relating to bacteria is significantly less understood. Nonetheless, Staphylococcus aureus appears to have at least five protein kinases, which would contribute to the manifestation of the antibiotic-resistance phenotype.
  • MIC minimal-inhibitory concentration
  • the MIC of oxacillin against the resistant MRSA strain NRS70 is 32 ⁇ g/mL.
  • Several inhibitors exhibited remarkable activity in lowering the MIC of oxacillin (Table 1) at the 20 ⁇ level.
  • the invention provides compounds of Formula A:
  • X is O, CH2, S, SO2, or a direct bond
  • each R 1 is independently CN, SO2CH3, CONH2, CO2H, C0 2 (alkyl), CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or S02-(pyridinyl);
  • n 1, 2, 3, or 4;
  • n 1, 2, 3, 4, or 5;
  • each R 2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, C0 2 Me, NO2, OCF 3 , or SCF 3 , or two R 2 groups form an ortho-fused methylenedioxy; and
  • each R 3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, C0 2 Me, NO2, OCF 3 , or SCF 3 , or two R 3 groups form an ortho-fused methylenedioxy;
  • R 1 a particular R 1 can be E or Z with respect to the phenyl ring to which the alkene of the R 1 is attached.
  • groups R 2 and R 3 are shown as attached at a variable location, they can be ortho, meta, or para to group X, or they can be excluded from locations ortho, meta, or para to group X.
  • the invention also provides compounds of Formula I:
  • X is O, CH2, S, SO2, or a direct bond
  • R 1 is CN, SO2CH3, CONH2, CO2H, CC Me, CO(aryl), pyridinyl, thiophi
  • n 1, 2, 3, or 4;
  • n 1, 2, 3, 4, or 5;
  • each R 2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, CC Me, NO2, OCF 3 , or SCF 3 , or two R 2 groups form an ortho-fused methylenedioxy; and
  • each R 3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NR a R b wherein R a and R b are each independently H or alkyl, C ⁇ CH, OH, CO2H, C0 2 Me, NO2, OCF3, or SCF3, or two R 3 groups form an ortho-fused methylenedioxy;
  • the group R 1 can be E or Z with respect to the phenyl ring to which the alkene of R 1 is attached. In certain specific embodiments, the group R 1 is in the Z configuration, and therefore the CN is in the Z configuration.
  • One specific value for X is O. Another specific value for X is CH2. Another specific value for X is S. Another specific value for X is SO2. Another specific value for X is a direct bond.
  • R 1 examples include CN, SO2CH3, CONH2, CO2H, and C0 2 Me.
  • Another specific value of R 1 is CO(aryl), for example, CO(phenyl), wherein phenyl is optionally substituted with one or more substituents, such as methyl, halo, methoxy, amino, nitro, CN, CF3, and the like, or a substituent as described herein.
  • Another specific value of R 1 is CO(4-MePh).
  • R 1 When R 1 is pyridyl, the pyridyl and be a 2-pyridyl, 3-pyridyl, or 4-pyridyl.
  • R 1 When R 1 is thiophenyl, the thiophenyl can be a 2- thiophenyl or a 3- thiophenyl.
  • R 1 When R 1 is benzothiazolyl, R 1 can be 2- benzothiazolyl, 4- benzothiazolyl, 5- benzothiazolyl, 6- benzothiazolyl, or 7- benzothiazolyl.
  • R 1 when R 1 is CO-cycloalkyl are CO-cyclopropyl, CO-cyclobutyl,
  • R 1 when R 1 is S02-(pyridinyl) are S02-(2-pyridinyl), S02-(3- pyridinyl), and SC -(4-pyridinyl).
  • m can be 1. In another embodiment, m can be 2. In yet another embodiment, m can be 3. In a further embodiment, m can be 4.
  • n can be 1. In another embodiment, n can be 2. In yet another embodiment, n can be 3. In a further embodiment, n can be 4. In yet a further embodiment, n can be 5.
  • R 2 Specific values for R 2 include H, C ⁇ CH, OH, C0 2 H, CC Me, NO2, OCF 3 , and SCF 3 .
  • R 2 can also be halo, for example, F, CI, Br, or I.
  • R 2 can also be alkyl, for example, (Ci-C6)alkyl, optionally branched. Examples include methyl, ethyl, propyl, seopropyl, wo-propyl, seobutyl, fert-butyl, and the like.
  • R 2 can also be alkoxy, for example, (C1-C6) alkoxy, optionally branched. Examples include methoxy, ethoxy, propoxy, seopropoxy, wo-propoxy, seobutoxy, fert-butoxy, and the like.
  • R 2 can also be heteroaryl, for example, thiophenyl, furanyl,benzothiazolyl, pyridyl, or pyrimidinyl, substituted on the heteroaryl at any available valency.
  • R 2 can also be heterocycle, for example, N-morpholinyl, tetrahydropyranyl,
  • two R 2 groups form an ortho-fused methylenedioxy.
  • R 3 include H, C ⁇ CH, OH, CO2H, C0 2 Me, ⁇ 2, OCF 3 , and SCF 3 .
  • R 3 can also be halo, for example, F, CI, Br, or I.
  • R 3 can also be alkyl, for example, (Ci-C6)alkyl, optionally branched. Examples include methyl, ethyl, propyl, seopropyl, wo-propyl, seobutyl, fert-butyl, and the like.
  • R 3 can also be alkoxy, for example, (C1-C6) alkoxy, optionally branched. Examples include methoxy, ethoxy, propoxy, seopropoxy, wo-propoxy, seobutoxy, fert-butoxy, and the like.
  • R 3 can also be heteroaryl, for example, thiophenyl, furanyl,benzothiazolyl, pyridyl, or pyrimidinyl, substituted on the heteroaryl at any available valency.
  • R 3 can also be heterocycle, for example, N-morpholinyl, tetrahydropyranyl,
  • two R 3 groups form an ortho-fused methylenedioxy.
  • a group or specific value of X, R 1 , R 2 , or R 3 is excluded from various embodiments of the invention.
  • the invention also provides the compounds illustrated in Figure 1. In some
  • compounds 81, 82, 85-90, 93-96, and 98-99 are excluded from such
  • preparation of the compounds and formulas described herein, and modifications thereof, can be made according to organic synthesis techniques known to those of skill in the art and/or according to the synthetic schemes provided herein.
  • synthesis of a subject compound can begin with commercially available chemicals, from compounds described in the chemical literature, or from products of the reactions and methods described herein.
  • Commercially available compounds may be obtained from standard commercial sources including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), ICN Biomedicals, Inc. (Costa Mesa, CA), Lancaster Synthesis (Windham, NH), Spectrum Quality Product, Inc. (New
  • reaction times and conditions are intended to be approximate, e.g., taking place at about atmospheric pressure within a temperature range of about -10 °C to about 110 °C over a period of about 1 to about 24 hours; reactions left to run overnight average a period of about 16 hours.
  • R 1 groups of Formulas A and I are installed by condensation of the appropriate disubstituted methylene group with the relevant aldehyde.
  • the disubstituted methylene groups are commercially available or can be prepared by standard synthetic techniques known to those of skill in the art.
  • compound 35-b is prepared using NC-CH2-CO(cyclopropyl)and compound 36-b is prepared using NC-CH2-SC (2-pyridyl).
  • a pharmaceutical composition comprises one or more compounds described herein and one or more antibiotic or antiseptic agents.
  • suitable active agents include penicillins, cephalosporins, carbacephems, cephamycins, carbapenems, monobactams, aminoglycosides, glycopeptides, quinolones, tetracyclines, macrolides, and fluoroquinolones.
  • Suitable antiseptic agents include iodine, silver, copper, chlorhexidine, polyhexanide and other biguanides, chitosan, acetic acid, and hydrogen peroxide. These agents may be incorporated as part of the same pharmaceutical composition or may be administered separately (concurrently or sequentially).
  • the pharmaceutical compositions may also contain anti-inflammatory drugs such as steroids and macrolactam derivatives.
  • ⁇ - Lactam antibiotics are bactericidal, and can act by inhibiting the synthesis of the peptidoglycan layer of bacterial cell walls.
  • the peptidoglycan layer is important for cell wall structural integrity, especially in Gram-positive bacteria.
  • ⁇ -lactam antibiotics include, but are not limited to, benzathine penicillin, benzylpenicillin (penicillin G), phenoxymethylpenicillin (penicillin V), procaine penicillin, methicillin, ceftadizim, ceftaroline, oxacillin, nafcillin, cloxacillin, dicloxacillin, flucloxacillin, temocillin, amoxicillin, ampicillin, co-amoxiclav, azlocillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, cephalosporins, cephalexin, cephalothin, cefazolin, cefaclor, cefuroxime, cefamandole, cephamycins, cefotetan, cefoxitin, ceftriaxone, cefotaxime, cefpodoxime, cefixime, ceftazidime, cefepime, cefp
  • Administration of ⁇ -lactam antibiotics can be carried out in conjunction with administration of a compound described herein.
  • the administration of the ⁇ -lactam antibiotic can be concurrent or sequential with respect to the administration of a compound described herein.
  • a compound described herein can be administered to a subject having a bacterial infection, and then an antibiotic can be administered, its activity potentiated by the administration of a compound described herein.
  • the antibiotic can be a ⁇ - lactam antibiotic recited herein above.
  • the antibiotic can be clavulanate, sulbactam, tazobactam, avibactam, MK-7655, or NXL105.
  • Susceptible organisms generally include gram positive and gram negative, aerobic and anaerobic organisms whose growth can be inhibited by embodiments described herein.
  • Susceptible organisms include, but are not limited to, Staphylococcus, Lactobacillus,
  • Streptococcus Streptococcus agalactiae, Sarcina, S. pneumoniae, S. pyogenes, S. mutans, Escherichia, Enter obacter , Klebsiella, Pseudomonas, Pseudomonas aeruginosa, Acinetobacter, Proteus, Campylobacter , Citrobacter, Nisseria, Bacillus anthracis, Bacillus cereus, Bacillus subtilis, Bacteroides, Peptococcus, Clostridium, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Mycobacterium tuberculosis and similar organisms.
  • the compounds described herein can be used to prepare therapeutic pharmaceutical compositions, for example, by combining the compounds with a pharmaceutically acceptable diluent, excipient, or carrier.
  • the compounds may be added to a carrier in the form of a salt or solvate.
  • administration of the compounds as salts may be appropriate.
  • Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids that form a physiologically acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, a-ketoglutarate, and ⁇ - glycerophosphate.
  • Suitable inorganic salts may also be formed, including hydrochloride, halide, sulfate, nitrate, bicarbonate, and carbonate salts.
  • salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid to provide a physiologically acceptable ionic compound.
  • a sufficiently basic compound such as an amine
  • a suitable acid for example, a sufficiently basic compound such as an amine
  • Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example, calcium) salts of carboxylic acids can also be prepared by analogous methods.
  • the compounds of the formulas described herein can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms.
  • the forms can be specifically adapted to a chosen route of administration, e.g., oral or parenteral administration, by intravenous, intramuscular, topical or subcutaneous routes.
  • the compounds described herein may be systemically administered in combination with a pharmaceutically acceptable vehicle, such as an inert diluent or an assimilable edible carrier.
  • a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier.
  • compounds can be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the food of a patient's diet.
  • Compounds may also be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • Such compositions and preparations typically contain at least 0.1% of active compound.
  • compositions and preparations can vary and may conveniently be from about 0.5% to about 60%, about 1 % to about 25%, or about 2% to about 10%, of the weight of a given unit dosage form.
  • the amount of active compound in such therapeutically useful compositions can be such that an effective dosage level can be obtained.
  • the tablets, troches, pills, capsules, and the like may also contain one or more of the following: binders such as gum tragacanth, acacia, com starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as com starch, potato starch, alginic acid and the like; and a lubricant such as magnesium stearate.
  • a sweetening agent such as sucrose, fructose, lactose or aspartame; or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring, may be added.
  • a liquid carrier such as a vegetable oil or a polyethylene glycol.
  • Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like.
  • a syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propyl parabens as preservatives, a dye and flavoring such as cherry or orange flavor.
  • Any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed.
  • the active compound may be incorporated into sustained-release preparations and devices.
  • the active compound may be administered intravenously or intraperitoneally by infusion or injection.
  • Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant.
  • Dispersions can be prepared in glycerol, liquid polyethylene glycols, triacetin, or mixtures thereof, or in a pharmaceutically acceptable oil. Under ordinary conditions of storage and use, preparations may contain a preservative to prevent the growth of microorganisms.
  • compositions suitable for injection or infusion can include sterile aqueous solutions, dispersions, or sterile powders comprising the active ingredient adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes.
  • the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage.
  • the liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and/or antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers, or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by agents delaying absorption, for example, aluminum monostearate and/or gelatin.
  • Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, optionally followed by filter sterilization.
  • methods of preparation can include vacuum drying and freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the solution.
  • compounds may be applied in pure form, e.g., when they are liquids.
  • a dermatologically acceptable carrier which may be a solid, a liquid, a gel, or the like.
  • Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina, and the like.
  • Useful liquid carriers include water, dimethyl sulfoxide (DMSO), alcohols, glycols, or water-alcohol/glycol blends, in which a compound can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants.
  • DMSO dimethyl sulfoxide
  • alcohols alcohols
  • glycols glycols
  • water-alcohol/glycol blends in which a compound can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants.
  • Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use.
  • the resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using a pump-type or aerosol sprayer.
  • Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses, or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.
  • compositions for delivering active agents to the skin are known to the art; for example, see U.S. Patent Nos. 4,992,478 (Geria), 4,820,508 (Wortzman), 4,608,392 (Jacquet et al), and 4,559,157 (Smith et al).
  • Such dermatological compositions can be used in combinations with the compounds described herein where an ingredient of such compositions can optionally be replaced by a compound described herein, or a compound described herein can be added to the composition.
  • Useful dosages of the compounds described herein can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Patent No. 4,938,949 (Borch et al.).
  • the amount of a compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient, and will be ultimately at the discretion of an attendant physician or clinician.
  • a suitable dose will be in the range of from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg/kg/day, most preferably in the range of 15 to 60 mg/kg/day.
  • the invention provides a composition comprising a compound of the invention formulated in such a unit dosage form.
  • the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day.
  • the sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.
  • the invention provides therapeutic methods of treating a bacterial infection in a mammal, which involve administering to a mammal having a bacterial infection an effective amount of a compound or composition described herein.
  • a mammal includes a primate, human, rodent, canine, feline, bovine, ovine, equine, swine, caprine, bovine and the like.
  • the ability of a compound of the invention to treat a bacterial infection may be determined by using assays well known to the art.
  • the invention also provides a kit comprising a packaging containing one or more doses of a first pharmaceutical formulation comprising a compound described herein or a
  • the first dose of the first pharmaceutical formulation comprises a loading dose of a compound described herein.
  • the first dose of the second pharmaceutical formulation comprises a loading dose of an antibiotic.
  • kits independently be in any dosage form, e.g. tablets, capsules, solutions, creams, etc. and packaged within any of the standard types of pharmaceutical packaging materials, e.g. bottles, blister- packs, IV bags, syringes, etc., that may themselves be contained within an outer packaging material such as a paper/cardboard box.
  • the kit further comprises one or more of culture media, culture plates, PCR primers, test strips, and/or stains for identifying the infective agent.
  • Thin-layer chromatography was done on EMD Millipore (Billerica, MA, U.S.A.) 0.25 mm silica gel 60 F254 plates. Column chromatography was done either manually using silica gel 60, 230-400 mesh (40-63 ⁇ particle size) purchased from Sigma- Aldrich Chemical Co., or on a Teledyne Combiflash Rf 200i automated chromatography system (Teledyne Isco, Lincoln, NE, U.S.A.) using disposable silica gel columns.
  • MIC Minimal-inhibitory concentration determination. Determination of MICs was carried out by the microdilution method cation-adjusted in Mueller-Hinton II Broth (CAMHB II, BBL) in accordance with the protocols of CLSI ⁇ Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Second Informational Supplement. CLSI document M100-S22. Clinical and Laboratory Standards Institute. Wayne, PA). A final bacterial inoculum of 5 ⁇ 10 5 CFU/rnL was used, and the results were recorded after incubation for 16-20 h at 37 °C.
  • the MIC values of 100 compounds were evaluated. Numerous compounds were evaluated at three or more different concentrations, typically: 0 (control), 2, and 20 ⁇ . If the compound exhibited inherent antibacterial activity at 20 ⁇ against the S. aureus strain
  • the compound was reevaluated at lower concentrations for the particular strain (e.g., 0, 0.1, and 1 ⁇ , or 0, 5, and 10 ⁇ ) to eliminate interference with the assay.
  • the particular strain e.g., 0, 0.1, and 1 ⁇ , or 0, 5, and 10 ⁇
  • Several compounds showed a greater than 2-fold decrease relative to the control.
  • ESP388 Ri S0 2 CH 3 , C0 2 Et (in place of CN; i.e., a compound
  • Potentiators 1 to 11 and ESP388 were evaluated in greater depth. The compounds at 20, 16, 12 and 8 ⁇ were evaluated as potentiators and as antibiotics. MRSA activity was tested against 7 different MRSA strains: NRS70, MRSA252 (USA200), NRS 123 (USA400), NRS382 (USA100), NRS384 (USA300), NRS1 (Vancomycin-Intermediate Staphylococcus aureus - VISA) and NRS 119 (a linezolid-Resistant Staphylococcus aureus).
  • compositions illustrate representative pharmaceutical dosage forms that may be used for the therapeutic or prophylactic administration of a compound of a formula described herein, a compound specifically disclosed herein, or a pharmaceutically acceptable salt or solvate thereof (hereinafter referred to as 'Compound X'):
  • Polyethylene glycol 400 200.0 0.1 N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5)
  • compositions may be prepared by conventional procedures well known in the pharmaceutical art. It will be appreciated that the above pharmaceutical compositions may be varied according to well-known pharmaceutical techniques to accommodate differing amounts and types of active ingredient 'Compound X'. Aerosol formulation (vi) may be used in conjunction with a standard, metered dose aerosol dispenser. Additionally, the specific ingredients and proportions are for illustrative purposes. Ingredients may be exchanged for suitable equivalents and proportions may be varied, according to the desired properties of the dosage form of interest. While specific embodiments have been described above with reference to the disclosed embodiments and examples, such embodiments are only illustrative and do not limit the scope of the invention. Changes and modifications can be made in accordance with ordinary skill in the art without departing from the invention in its broader aspects as defined in the following claims.

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Abstract

Proteins of methicillin-resistant Staphylococcus aureus (MRSA), an antibiotic sensor/signal transducer, are phosphorylated on exposure to β-lactam antibiotics. This event is critical for the onset of the biochemical events that unleash induction of antibiotic resistance. The phosphorylation and the antibiotic-resistance phenotype can be abrogated in the presence of inhibitors described herein that restore susceptibility of the organism to β- lactam antibiotics. The invention thus provides compounds and methods for abrogating antibiotic resistance to β-lactam antibiotics and for treating infections causes by antibiotics prone to developing resistance by potentiating β-lactam antibiotics.

Description

POTENTIATORS OF BETA-LACTAM ANTIBIOTICS
AND COMBINATION THERAPY
RELATED APPLICATIONS
This application claims priority under 35 U. S.C. § 119(e) to U. S. Provisional Patent Application No. 62/267,780, filed December 15, 2015, which is incorporated herein by reference.
GOVERNMENT SUPPORT
This invention was made with government support under Grant No. All 04987 awarded by the National Institutes of Health. The government has certain rights in the invention.
BACKGROUND OF THE INVENTION
Staphylococcus aureus is a Gram-positive bacterium commonly found on the skin and in moist areas, such as the nasal cavity, yet it is often broadly resistant to many antibiotics, β- Lactam antibiotics were the drugs of choice for treatment of infection by S. aureus, but a variant of this organism, methicillin-resistant Staphylococcus aureus (MRS A) emerged in 1961, which exhibited resistance to the entire class of β-lactams. This organism has been a global clinical problem for over half a century. The molecular basis for the broad resistance of MRS A to β- lactams, which is incidentally inducible, was traced to a set of genes within the bla and mec operons. The BlaRl (or the cognate MecRl) protein is a β-lactam antibiotic sensor/signal transducer, which communicates the presence of the antibiotic in the milieu to the cytoplasm in a process that is largely not understood (Staude et al., Biochemistry 2015, 54, 1600-1610). Signal transduction leads to activation of the cytoplasmic domain of BlaRl (or MecRl), a zinc protease, which turns over the gene repressor Blal (or Mecl) in derepressing transcriptional events that result in expression of antibiotic-resistance determinants, the class A β-lactamase PCI and/or the penicillin-binding protein 2a (PBP2a) (Llarrull and Mobashery, Biochemistry 2012, 51, 4642-4649).
An intriguing aspect of this system is its inducibility. Upon exposure to the antibiotic, the organism mobilizes. Once the antibiotic challenge is withdrawn, the system reverses itself. It was argued that when the signal for the presence of the antibiotic transduces to the cytoplasmic domain, the BlaRl protein undergoes autoproteolysis, which unleashes the activity of the protease domain in degradation of the gene repressor Blal. We have found that this autoproteolytic processing takes place in the absence of antibiotic as well. Therefore, proteolysis may lead to turnover of BlaRl itself as an event in the reversal of induction. What is needed is the identification of what accounts for activation of the cytoplasmic domain toward degradation of Blal in manifestation of the antibiotic-resistance response. Such identification could provide needed methods for reducing, preventing, or otherwise abrogating resistance to β- lactam antibiotics.
SUMMARY
One or more proteins of methicillin-resistant Staphylococcus aureus (MRSA), an antibiotic sensor/signal transducer, are phosphorylated on exposure to β-lactam antibiotics. This event is critical for the onset of the biochemical events that unleash induction of antibiotic resistance. The phosphorylation and the antibiotic-resistance phenotype are abrogated in the presence of novel inhibitors that restore susceptibility of the organism to β-lactam antibiotics. The invention provides compounds, compositions, and methods for reducing, preventing, overcoming, and/or abrogating resistance to β-lactam antibiotics, and methods of treating bacterial infections caused by antibiotic resistant bacteria, particularly bacteria that can develop resistance to β-lactam antibiotics. Accordingly, the invention provides compounds that potentiate the antibacterial activity of β-lactam antibiotics toward β-lactam-resistant antibiotics.
The invention therefore provides compositions and methods for increasing the sensitivity of bacterial pathogens to antibiotics (e.g., potentiating the antibiotics), including β-lactam antibiotics. In one embodiment, the invention provides a method for increasing the sensitivity of bacterial pathogens to β-lactam antibiotics by contacting the bacterial pathogen with one or more compounds described herein. In some embodiments, the bacterial pathogen is MSRA. In other embodiments, the bacterial pathogen is Enterococcus faecalis or other bacterial pathogen described herein.
The invention also provides compositions and methods for increasing the susceptibility of Gram positive or Gram negative pathogens to β-lactam antibiotics. Various embodiments provide pharmaceutical compositions, therapeutic formulations, product combination, or kits for use against MRSA infections comprising a compound described herein and one or more β- lactam antibiotics. The compounds and methods can be used for inhibiting the growth of bacteria, for example, Staphylococcus aureus. In some embodiments, the Staphylococcus aureus is resistant to, or sensitive to, methicillin, other β-lactams, macrolides, lincosamides, aminoglycosides, or a combination thereof. Thus, the invention further provides methods for increasing the sensitivity of Staphylococcus aureus to methicillin, other β-lactams, macrolides, lincosamides, or aminoglycosides. The methods can include administering an effective amount of a compound, a pair of compounds, or composition described herein.
The compounds described herein include a compound of Formula I:
Figure imgf000005_0001
(I)
wherein
X is O, CH2, S, SO2, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO2H, CC Me, CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or S02-(pyridinyl);
m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
each R2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, CC Me, NO2, OCF3, or SCF3, or two R2 groups form an ortho-fused methylenedioxy; and
each R3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
In one embodiment, X is O. In another embodiment, X is CH2. In yet another embodiment, X is S. In yet a further embodiment, X is a direct bond. In yet a further embodiment, X is SO2.
In one embodiment, R1 is CN or SO2CH3. In some embodiments, R2 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused
methylenedioxy. In various embodiments, R3 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy.
In some embodiments, X is O, CH2, S, or a direct bond, R1 is CN or SO2CH3, m is 1 or 2, and n is 1 or 2. In such embodiments, R2 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy; and R3 is CI, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R3 groups form an ortho-fused
methylenedioxy.
The compounds of Formula I include the various compounds of Formulas II and III, including their sub-Formulas II-A, II-B, II-C, II-D, III, III-A, and III-B. Accordingly, the invention provides a compound of Formula I wherein the compound is a compound of Formula II:
Figure imgf000006_0001
wherein
X is O, CH2, S. or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
R2 is H, F. OMe, OEt, OCF3, SCF3. N-morpholinyl, or pyridinyl. or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, N-mo holinyl, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
The invention also provides compounds of Formula II wherein the compound is a compound of Formula II-A:
Figure imgf000006_0002
wherein
R1 is CN. SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof. The invention also provides compounds of Formula II wherein the compound is a compound of Formula II-B:
Figure imgf000007_0001
wherein
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
The invention also provides compounds of Formula II wherein the compound is a compound of Formula II-C:
Figure imgf000007_0002
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof. The invention also provides compounds of Formula II wherein the compound is a compound of Formula II-D:
Figure imgf000008_0001
wherein
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
The invention also provides compounds of Formula I wherein the compound is a compound of Formula III:
Figure imgf000008_0002
wherein
X is O, CH2, S, or a direct bond;
R1 is CN. SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S02-(2- pyridinyl);
m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, SCF3, or N-morpholinyl; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof. The invention further provides compounds of Formula III wherein the compound is a compound of Formula III-A:
Figure imgf000009_0001
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S02-(2- pyridinyl);
m is 1 or 2; and
R2 is H, F, OMe, OEt, OCF3, SCF3, or N-morpholinyl;
or a salt or solvate thereof.
The invention yet further provides compounds of Formula III wherein the compound is a compound of Formula III-B:
Figure imgf000009_0002
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(aryl), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); n is 2;
R2 is F; and
R3 is CI;
or a salt or solvate thereof.
The invention further provides the compounds illustrated in Figure 3, or a salt or solvate thereof, for example, one or more compounds of Formulas I-III as illustrated in Figure 3. Also provided are compositions comprising a compound of any one of Formulas I-III and/or Figure 3 in combination with a pharmaceutically acceptable diluent, excipient, or carrier. Further provided are compositions comprising a compound of any one of Formulas I-III and/or Figure 3 in combination with a β-lactam antibiotic. In various embodiments, β-lactam antibiotic is ceftadizim, ceftaroline, ceftazidime, meropenem, oxacillin, or penicillin, or another antibiotic recited herein. The invention further provides a method to reverse the methicillin-resistant phenotype in bacteria comprising contacting methicillin-resistant Staphylococcus aureus (MRSA) with an effective amount of a compound described herein, thereby rendering MRSA susceptible to β- lactam antibiotics.
The invention yet further provides a method to inhibit or kill methicillin-resistant
Staphylococcus aureus (MRSA) comprising contacting the MRSA with an amount of a compound described herein effective to reverse the methicillin-resistant phenotype, and contacting the MRSA with an effective antibacterial amount of a β-lactam antibiotic.
In another embodiment, the invention provides a method to reduce or attenuate the minimum inhibitory concentration (MIC) of a β-lactam antibiotic comprising contacting a bacterium with an effective amount of a compound described herein in combination with contacting the bacterium with a β-lactam antibiotic. The invention therefore provides for the use of a compound described herein for preparing a medicament to treat a bacterial infection. The bacterial infection can be, for example, a methicillin-resistant Staphylococcus aureus (MRSA) infection. The administration of the compound described herein can be concurrent or sequential with an antibiotic, for example, a β-lactam antibiotic, including the specific β-lactam antibiotics described herein.
Further embodiments relate to methods of ameliorating and/or treating a bacterial infection that can include administering to a subject suffering from the bacterial infection an effective amount of one or more compounds of Formulas I-III, or a pharmaceutical composition that includes one or more compounds of Formulas I-III, or a pharmaceutically acceptable salt thereof. Other embodiments described herein relate to using one or more compounds of Formulas I-III in the manufacture of a medicament for ameliorating and/or treating a bacterial infection. Still other embodiments described herein relate to compounds of Formulas I-III that can be used for ameliorating and/or treating a bacterial infection. Other embodiments relate to methods of ameliorating and/or treating a bacterial infection that can include administering to a patient infected with the bacterial infection an effective amount of one or more compounds of Formulas I-III. Some embodiments described herein relate to methods of inhibiting the replication of a bacteria that can include administering to a patient infected with the bacteria an effective amount of one or more compounds of Formulas I-III. In one embodiment, the bacterial infection can be an S. aureus infection, for example, a MRSA infection.
The invention thus provides novel compounds of Formulas I-III, intermediates for the synthesis of compounds of Formulas I-III, as well as methods of preparing compounds of Formulas I-III. The invention also provides compounds of Formulas I-III that are useful as intermediates for the synthesis of other useful compounds. The invention provides for the use of the compounds and compositions described herein in medical therapy. The compounds of Formulas I-III can be used in the manufacture of medicaments useful for the treatment of bacterial infections in a mammal, such as a human. Compositions and medicaments described herein can include a pharmaceutically acceptable diluent, excipient, or carrier.
BRIEF DESCRIPTION OF THE DRAWINGS
The following drawings form part of the specification and are included to further demonstrate certain embodiments or various aspects of the invention. In some instances, embodiments of the invention can be best understood by referring to the accompanying drawings in combination with the detailed description presented herein. The description and accompanying drawings may highlight a certain specific example, or a certain aspect of the invention. However, one skilled in the art will understand that portions of the example or aspect may be used in combination with other examples or aspects of the invention.
Figure 1. Chemical structures of examples of compounds of the formulas described herein, according do certain embodiments.
DETAILED DESCRIPTION
Antimicrobial resistance is one of the most serious threats to global public health today. β-Lactam antibiotics were the preferred antibiotics for treatment of infections by S. aureus, but emergence of Methicillin-resistant Staphylococcus aureus (MRS A) in 1961 made these drugs obsolete within a short time. In response to exposure to β-lactam antibiotics, MRSA exhibits phosphorylation of certain proteins. Interference with phosphorylation or other critical pathways reverses the antibiotic-resistance phenotype. We describe herein a set of molecules that are capable of this phenotype reversal, restoring sensitivity to the organism against β-lactam antibiotics.
Protein of methicillin-resistant Staphylococcus aureus (MRSA), an antibiotic
sensor/signal transducer, are phosphorylated on exposure to β-lactam antibiotics. This event is critical for the onset of the biochemical events that unleash induction of antibiotic resistance. The phosphorylation and the antibiotic-resistance phenotype may be abrogated in the presence of novel inhibitors described herein, which inhibitors restore susceptibility of the organism to β- lactam antibiotics. The invention thus provides compounds and methods for abrogating antibiotic resistance to β-lactam antibiotics, thereby potentiating the activity of a β-lactam antibiotic. Definitions
The following definitions are included to provide a clear and consistent understanding of the specification and claims. As used herein, the recited terms have the following meanings. All other terms and phrases used in this specification have their ordinary meanings as one of skill in the art would understand. Such ordinary meanings may be obtained by reference to technical dictionaries, such as Hawley 's Condensed Chemical Dictionary 14th Edition, by R.J. Lewis, John Wiley & Sons, New York, N.Y., 2001.
References in the specification to "one embodiment", "an embodiment", etc., indicate that the embodiment described may include a particular aspect, feature, structure, moiety, or characteristic, but not every embodiment necessarily includes that aspect, feature, structure, moiety, or characteristic. Moreover, such phrases may, but do not necessarily, refer to the same embodiment referred to in other portions of the specification. Further, when a particular aspect, feature, structure, moiety, or characteristic is described in connection with an embodiment, it is within the knowledge of one skilled in the art to affect or connect such aspect, feature, structure, moiety, or characteristic with other embodiments, whether or not explicitly described.
The singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a compound" includes a plurality of such compounds, so that a compound X includes a plurality of compounds X. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for the use of exclusive terminology, such as "solely," "only," and the like, in connection with any element described herein, and/or the recitation of claim elements or use of "negative" limitations.
The term "and/or" means any one of the items, any combination of the items, or all of the items with which this term is associated. The phrases "one or more" and "at least one" are readily understood by one of skill in the art, particularly when read in context of its usage. For example, the phrase can mean one, two, three, four, five, six, ten, 100, or any upper limit approximately 10, 100, or 1000 times higher than a recited lower limit. For example, one or more substituents on a phenyl ring refers to one to five, or one to four, for example if the phenyl ring is disubstituted.
The term "about" can refer to a variation of ± 5%, ± 10%, ± 20%, or ± 25% of the value specified. For example, "about 50" percent can in some embodiments carry a variation from 45 to 55 percent. For integer ranges, the term "about" can include one or two integers greater than and/or less than a recited integer at each end of the range. Unless indicated otherwise herein, the term "about" is intended to include values, e.g., weight percentages, proximate to the recited range that are equivalent in terms of the functionality of the individual ingredient, the composition, or the embodiment. The term about can also modify the end-points of a recited range as discussed above in this paragraph.
As will be understood by the skilled artisan, all numbers, including those expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth, are approximations and are understood as being optionally modified in all instances by the term "about. " These values can vary depending upon the desired properties sought to be obtained by those skilled in the art utilizing the teachings of the descriptions herein. It is also understood that such values inherently contain variability necessarily resulting from the standard deviations found in their respective testing measurements.
As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges recited herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof, as well as the individual values making up the range, particularly integer values. A recited range (e.g., weight percentages or carbon groups) includes each specific value, integer, decimal, or identity within the range. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, or tenths. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art, all language such as "up to", "at least", "greater than", "less than", "more than", "or more", and the like, include the number recited and such terms refer to ranges that can be subsequently broken down into subranges as discussed above. In the same manner, all ratios recited herein also include all sub- ratios falling within the broader ratio. Accordingly, specific values recited for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for radicals and substituents.
One skilled in the art will also readily recognize that where members are grouped together in a common manner, such as in a Markush group, the invention encompasses not only the entire group listed as a whole, but each member of the group individually and all possible subgroups of the main group. Additionally, for all purposes, the invention encompasses not only the main group, but also the main group absent one or more of the group members. The invention therefore envisages the explicit exclusion of any one or more of members of a recited group. Accordingly, provisos may apply to any of the disclosed categories or embodiments whereby any one or more of the recited elements, species, or embodiments, may be excluded from such categories or embodiments, for example, for use in an explicit negative limitation. The term "alkyl" refers to a straight- or branched-chain alkyl group having from 1 to about 20 carbon atoms in the chain. For example, the alkyl group can be a (Ci-C2o)alkyl, a (Ci- Ci2)alkyl, (Ci-C8)alkyl, (Ci-C6)alkyl, or (Ci-C4)alkyl. Examples of alkyl groups include methyl (Me), ethyl (Et), ^-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (t- u), pentyl, isopentyl, fert-pentyl, hexyl, isohexyl, and groups that in light of the ordinary skill in the art and the teachings provided herein would be considered equivalent to any one of the foregoing examples. Alkyl groups can be optionally substituted or unsubstituted, and optionally partially unsaturated, such as in an alkenyl group.
The term "cycloalkyl" refers to cyclic alkyl groups of, for example, from 3 to 10 carbon atoms having a single cyclic ring or multiple condensed rings. Cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantyl, pinenyl, and the like. The cycloalkyl group can be monovalent or divalent, and can be optionally substituted, for example, by one or more alkyl groups. The cycloalkyl group can optionally include one or more cites of unsaturation, for example, the cycloalkyl group can include one or more carbon-carbon double bonds, such as, for example, 1-cyclopent-l-enyl, l-cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, l-cyclohex-3-enyl, and the like.
The term "alkoxy" refers to the group alky 1-0-, where alkyl is as defined herein.
Preferred alkoxy groups include, e.g., methoxy, ethoxy, w-propoxy, wo-propoxy, w-butoxy, fert-butoxy, sec-butoxy, w-pentoxy, w-hexoxy, 1,2-dimethylbutoxy, and the like.
The term "cycloalkyl" refers to cyclic alkyl groups of, for example, from 3 to 10 carbon atoms having a single cyclic ring or multiple condensed rings. Cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantyl, pinenyl, and the like. The cycloalkyl group can be monovalent or divalent, and can be optionally substituted, for example, by one or more alkyl groups. The cycloalkyl group can optionally include one or more cites of unsaturation, for example, the cycloalkyl group can include one or more carbon-carbon double bonds, such as, for example, 1-cyclopent-l-enyl, l-cyclopent-2-enyl, l-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, l-cyclohex-3-enyl, and the like.
The term "alkoxy" refers to the group alky 1-0-, where alkyl is as defined herein.
Preferred alkoxy groups include, e.g., methoxy, ethoxy, w-propoxy, wo-propoxy, w-butoxy, fert-butoxy, sec-butoxy, w-pentoxy, w-hexoxy, 1,2-dimethylbutoxy, and the like.
The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic ring system containing one, two, or three aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring. The heteroaryl can be unsubstituted or substituted, for example, with one or more, and in particular one to three, substituents, as described in the definition of "substituted" . Typical heteroaryl groups contain 2-20 carbon atoms in the ring skeleton in addition to the one or more heteroatoms.
Examples of heteroaryl groups include, but are not limited to, 2H-pyrrolyl, 3H-indolyl,
4H-quinolizinyl, acridinyl, benzo[b]thienyl, benzothiazolyl, β-carbolinyl, carbazolyl, chromenyl, cinnolinyl, dibenzo[b,d]furanyl, furazanyl, furyl, imidazolyl, imidizolyl, indazolyl, indolisinyl, indolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxazolyl, perimidinyl, phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, thiadiazolyl, thianthrenyl, thiazolyl, thienyl, triazolyl, tetrazolyl, and xanthenyl. In one embodiment the term "heteroaryl" denotes a monocyclic aromatic ring containing five or six ring atoms containing carbon and 1, 2, 3, or 4 heteroatoms independently selected from non-peroxide oxygen, sulfur, and N(Z) wherein Z is absent or is H, O, alkyl, aryl, or (Ci-C6)alkylaryl. In some embodiments, heteroaryl denotes an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benz-derivative or one derived by fusing a propylene, trimethylene, or tetramethylene diradical thereto.
The heteroaryl can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, acetamido, acetoxy, acetyl, benzamido, benzenesulfinyl, benzenesulfonamido, benzenesulfonyl, benzenesulfonylamino, benzoyl, benzoylamino, benzoyloxy, benzyl, benzyloxy,
benzyloxycarbonyl, benzylthio, carbamoyl, carbamate, isocyannato, sulfamoyl, sulfinamoyl, sulfino, sulfo, sulfoamino, thiosulfo, NRxRy and/or COORx, wherein each Rx and Ry are independently H, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl, or hydroxy. For example, the nitrogen of any indolyl ring can be N-substituted to provide an N-alkyl, N-methyl, or N-protecting group indolyl compound. A heteroaryl can also be substituted with a substituent as described in the substituents definition below.
The term "heterocycle" or "heterocyclyl" refers to a saturated or partially unsaturated ring system, containing at least one heteroatom selected from the group oxygen, nitrogen, and sulfur, and optionally substituted with alkyl, or C(=0)ORb, wherein Rb is hydrogen or alkyl. Typically heterocycle is a monocyclic, bicyclic, or tricyclic group containing one or more heteroatoms selected from the group oxygen, nitrogen, and sulfur. A heterocycle group also can contain an oxo group (=0) attached to the ring. Non-limiting examples of heterocycle groups include 1,3-dihydrobenzofuran, 1 ,3-dioxolane, 1 ,4-dioxane, 1 ,4-dithiane, 2H-pyran, 2- pyrazoline, 4H-pyran, chromanyl, imidazolidinyl, imidazolinyl, indolinyl, isochromanyl, isoindolinyl, morpholine, piperazinyl, piperidine, piperidyl, pyrazolidine, pyrazolidinyl, pyrazolinyl, pyrrolidine, pyrroline, quinuclidine, and thiomorpholine. The heterocycle can optionally be a divalent radical, thereby providing a heterocyclene.
The heterocycle can optionally be substituted with one or more alkyl, alkenyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, imino, alkylamino, acylamino, nitro, trifluoromethyl, trifiuoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, cyano, acetamido, acetoxy, acetyl, benzamido, benzenesulfinyl, benzenesulfonamido, benzenesulfonyl, benzenesulfonylamino, benzoyl, benzoylamino, benzoyloxy, benzyl, benzyloxy,
benzyloxy carbonyl, benzylthio, carbamoyl, carbamate, isocyannato, sulfamoyl, sulfinamoyl, sulfino, sulfo, sulfoamino, thiosulfo, NRxRy and/or COORx, wherein each Rx and Ry are independently Η, alkyl, alkenyl, aryl, heteroaryl, heterocycle, cycloalkyl, or hydroxy. A heterocycle can also be substituted with a substituent as described in the substituents definition below.
Examples of nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containing heterocycles.
Protecting Groups. Compounds of the invention can further include one or more suitable protecting groups. The term "protecting group" refers to any group that, when bound to an sp- center, a hydroxyl, nitrogen, or other heteroatom prevents undesired reactions from occurring at this group and that can be removed by conventional chemical or enzymatic steps to reestablish the 'unprotected' moiety, such as an alkyne, hydroxyl, nitrogen, or other heteroatom group. The particular removable group employed is often interchangeable with other groups in various synthetic routes. Certain removable protecting groups include conventional substituents such as, for example, allyl, benzyl, acetyl, chloroacetyl, thiobenzyl, benzylidine, phenacyl, methyl methoxy, silicon protecting groups ("silyl ethers") (e.g., trimethylsilyl (TMS), /-butyl- diphenylsilyl (TBDPS), triisopropylsilyl (TIPS), or /-butyldimethylsilyl (TBS)) and any other group that can be introduced chemically onto a hydroxyl or other moiety and later selectively removed either by chemical or enzymatic methods in mild conditions compatible with the nature of the product.
A large number of protecting groups and corresponding chemical cleavage reactions are described in Protective Groups in Organic Synthesis, Theodora W. Greene (John Wiley & Sons, Inc., New York, 1991, ISBN 0-471-62301-6) ("Greene", which is incorporated herein by reference in its entirety). Greene describes many nitrogen protecting groups, for example, amide-forming groups. In particular, see Chapter 1, Protecting Groups: An Overview, pages 1-20, Chapter 2, Hydroxyl Protecting Groups, pages 21-94, Chapter 4, Carboxyl Protecting Groups, pages 118-154, and Chapter 5, Carbonyl Protecting Groups, pages 155-184. See also Kocienski, Philip J. ; Protecting Groups (Georg Thieme Verlag Stuttgart, New York, 1994), which is incorporated herein by reference in its entirety. Some specific protecting groups that can be employed in conjunction with the methods of the invention are discussed below.
The term "halogen" refers to chlorine, fluorine, bromine or iodine. The term "halo" refers to chloro, fluoro, bromo or iodo.
As to any of the groups or "substituents" described herein (e.g., groups R1, R2, and R3), each can further include one or more (e.g., 1, 2, 3, 4, 5, or 6) substituents. It is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
The term "substituted" means that a specified group or moiety can bear one or more (e.g., 1, 2, 3, 4, 5, or 6) substituents. The term "unsubstituted" means that the specified group bears no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted by one or more substituents, and elements of the Formulas described herein can be optionally substituted. Where the term "substituted" is used to describe a structural system, the substitution is meant to occur at any valency-allowed position on the system. In cases where a specified moiety or group is not expressly noted as being optionally substituted or substituted with any specified substituent, it is understood that such a moiety or group is intended to be unsubstituted in some embodiments but can be substituted in other embodiments. In other words, the variables R1, R2, and R3 and their elements can be optionally substituted. In various embodiments, suitable substituent groups (e.g., on groups R1, R2, and R3 and/or their elements) include one or more of alkyl, alkenyl, alkynyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, aroyl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxy carbonyl, amino, alkylamino, dialkylamino, trifluoromethylthio, difluoromethyl, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, arylsulfinyl, arylsulfonyl, heteroarylsulfinyl, heteroarylsulfonyl, heterocyclesulfinyl, heterocyclesulfonyl, phosphate, sulfate, hydroxyl amine, hydroxyl (alkyl)amine, and/or cyano. Additionally, suitable substituent groups can be, e.g., -X, -R, -OH, -OR, -SR, -S", -NR2, -NR3, =NR, -CX3, -CN, -OCN, -SCN, -N=C=0, -NCS, -NO, -NO2, =N2, -N3, NC(=0)R, -C(=0)R, -C(=0)NRR, -S(=0)2H, -S(=0)2OH, -S(=0)2R, -OS(=0)2OR, -S(=0)2NHR, -S(=0)R, -C(=0)R, -C(=0)X, -C(S)R, -C(0)OR, -C(0)0\ -C(S)OR, -C(0)SR, -C(S)SR, -C(0)NRR, -C(S)NRR, or -C(NR)NRR, where each X is independently a halogen ("halo"): F, CI, Br, or I; and each R is independently H, alkyl, aryl, (aryl)alkyl (e.g., benzyl), heteroaryl, (heteroaryl)alkyl, heterocycle, heterocycle(alkyl), or a protecting group. As would be readily understood by one skilled in the art, when a substituent is keto (=0) or thioxo (=S), or the like, then two hydrogen atoms on the substituted atom are replaced. In certain embodiments, any one of the above groups can be included or excluded from a variable (e.g., groups R2 and R3) or from a group of substituents.
The term "pharmaceutically acceptable salts" refers to ionic compounds, wherein a parent non-ionic compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include conventional non-toxic salts and quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Non-toxic salts can include those derived from inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, phosphoric, nitric and the like. Salts prepared from organic acids can include those such as acetic, 2-acetoxybenzoic, ascorbic, behenic, benzenesulfonic, benzoic, citric, ethanesulfonic, ethane disulfonic, formic, fumaric, gentisinic, glucaronic, gluconic, glutamic, gly colic, hydroxymaleic, isethionic, isonicotinic, lactic, maleic, malic, mesylate or methanesulfonic, oxalic, pamoic (l, l '-methylene-bis-(2- hydroxy-3-naphthoate)), pantothenic, phenylacetic, propionic, salicylic, sulfanilic,
toluenesulfonic, stearic, succinic, tartaric, bitartaric, and the like. Certain compounds can form pharmaceutically acceptable salts with various amino acids. For a review on pharmaceutically acceptable salts, see, e.g., Berge et al, J. Pharm. Sci. 1977, 66(\), 1-19, which is incorporated herein by reference.
The pharmaceutically acceptable salts of the compounds described herein can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of many suitable salts are found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott, Williams & Wilkins, (2005).
The term "solvate" refers to a solid compound that has one or more solvent molecules associated with its solid structure. Solvates can form when a solid compound is crystallized from a solvent, wherein one or more solvent molecules become an integral part of the solid crystalline matrix. The compounds of the formulas described herein can be solvates, for example, ethanol solvates. Another type of a solvate is a hydrate. A "hydrate" likewise refers to a solid compound that has one or more water molecules intimately associated with its solid or crystalline structure at the molecular level. A hydrate is a specific type of a solvate. Hydrates can form when a compound is solidified or crystallized in water, wherein one or more water molecules become an integral part of the solid crystalline matrix. The compounds of the formulas described herein can be hydrates.
The term "diluent" refers to a pharmacologically inert substance that is nevertheless suitable for human consumption that serves as an excipient in the inventive dosage form. A diluent serves to dilute the API in the inventive dosage form, such that tablets of a typical size can be prepared incorporating a wide range of actual doses of the API.
The term "excipient" refers to an ingredient of the dosage form that is not medicinally active, but serves to dilute the API, assist in dispersion of the tablet in the patient's stomach, bind the tablet together, and serve other functions like stabilizing the API against decomposition.
The term "contacting" refers to the act of touching, making contact, or of bringing to immediate or close proximity, including at the cellular or molecular level, for example, to bring about a physiological reaction, a chemical reaction, or a physical change, e.g., in a solution, in a reaction mixture, in vitro, or in vivo (e.g., by administration to a patient).
An "effective amount" refers to an amount effective to treat a disease, disorder, and/or condition, or to bring about a recited effect. For example, an effective amount can be an amount effective to reduce the progression or severity of the condition or symptoms being treated. Determination of a therapeutically effective amount is well within the capacity of persons skilled in the art, especially in light of the detailed disclosure provided herein. The term "effective amount" is intended to include an amount of a compound described herein, or an amount of a combination of compounds described herein, e.g., that is effective to treat or prevent a disease or disorder, or to treat the symptoms of the disease or disorder, in a host. Thus, an "effective amount" generally means an amount that provides the desired effect. For example, the term "effective amount" can refer to an amount of compound or composition, which upon administration, is capable of reducing or preventing proliferation of a bacteria, reducing or preventing symptoms associated with a bacterial infection, reducing the likelihood of bacterial infection, or preventing bacterial infection. Typically, the subject is treated with an amount of a therapeutic composition sufficient to reduce a symptom of a disease or disorder, such as an infection, by at least about 25%, about 50%, about 75%, or about 90%.
The terms "treating", "treat" and "treatment" can include (i) preventing a disease, pathologic or medical condition from occurring (e.g., prophylaxis); (ii) inhibiting the disease, pathologic or medical condition or arresting its development; (iii) relieving the disease, pathologic or medical condition; and/or (iv) diminishing symptoms associated with the disease, pathologic or medical condition. Thus, the terms "treat", "treatment", and "treating" can extend to prophylaxis and can include prevent, prevention, preventing, lowering, stopping or reversing the progression or severity of the condition or symptoms being treated. As such, the term "treatment" can include medical, therapeutic, and/or prophylactic administration, as appropriate.
Treatments may be reactive, such as for combating an existing infection, or prophylactic, for preventing infection in an organism susceptible to infection. In some embodiments, compositions can be used to treat infections by drug-resistant strains of bacteria, for example MRSA (methicillin resistant S. aureus), MRSE (methicillin resistant S. epidermidis), PRSP (penicillin resistant S. pneumoniae), VIRSA (vancomycin intermittently resistant
Staphylococcus aureus) or VRE (vancomycin resistant Enter ococci). The term "drug-resistant" a condition where the bacteria are resistant to treatment with one or more conventional antibiotics, particularly β-lactam antibiotics. Accordingly, the invention provides a method for killing or inhibiting growth of gram positive bacteria comprising contacting gram positive bacteria with a compound or composition described herein, thereby killing or inhibiting the growth of the bacteria. The contacting can be performed in vivo in a human or animal, or in vitro, for example, in an assay. The gram positive bacteria can be of the genus Enterococcus or Staphylococcus. In certain embodiments, the bacteria is a drug-resistant strain of the genus Staphylococcus. In certain specific embodiments, the bacteria is a methicillin-resistant
Staphylococcus aureus (MRSA) strain.
In some embodiments, the bacterial infection may be due to Gram-positive bacteria, including, but not limited to, methicillin resistant Staphylococcus aureus (MRSA), community- acquired methicillin resistant Staphylococcus aureus (CAMRSA), vancomycin-intermediate- susceptible Staphylococcus aureus (VISA), methicillin-resistant coagulase-negative
staphylococci (MR-CoNS), vancomycin-intermediate-susceptible coagulase-negative staphylococci (VI-CoNS), methicillin susceptible Staphylococcus aureus (MSSA),
Streptococcus pneumoniae (including penicillin-resistant strains [PRSP]) and multi-drug resistant strains [MDRSP]), Streptococcus agalactiae, Streptococcus pyogenes and
Enterococcus faecalis. In particular embodiments, the bacterial infection may include, but is not limited to, complicated skin and skin structure infections (cSSSI); community acquired pneumonia (CAP); complicated intra-abdominal infections, such as, complicated appendicitis, peritonitis, complicated cholecystitis and complicated diverticulitis; uncomplicated and complicated urinary tract infections, such as, pyelonephritis; and respiratory and other nosocomial infections.
The term "infection" refers to the invasion of the host by germs (e.g., bacteria) that reproduce and multiply, causing disease by local cell injury, release of poisons, or germ- antibody reaction in the cells. The compounds and compositions described herein can be used to treat a gram positive bacterial infection, for example, in a mammal, such as a human.
The terms "inhibit", "inhibiting", and "inhibition" refer to the slowing, halting, or reversing the growth or progression of a disease, infection, condition, or group of cells. The inhibition can be greater than about 20%, 40%, 60%, 80%, 90%, 95%, or 99%, for example, compared to the growth or progression that occurs in the absence of the treatment or contacting.
Abrogation of Antibiotic Resistance by Small Molecules
Protein phosphorylation and its contribution to many regulatory events are widely known in eukaryotes. However, the same information relating to bacteria is significantly less understood. Nonetheless, Staphylococcus aureus appears to have at least five protein kinases, which would contribute to the manifestation of the antibiotic-resistance phenotype.
The minimal-inhibitory concentration (MIC) of oxacillin (a penicillin), meropenem, and ceftadizim toward various resistance S. aureus strains are shown in the table below.
Figure imgf000021_0001
We determined MICs (broth microdilution method) (CLSI, Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Second Informational Supplement. CLSI document M100-S22. Clinical and Laboratory Standards Institute; Wayne, PA) for 99 inhibitors described herein, in case some have antibacterial properties of their own, which could complicate analysis. Indeed, a few of these compounds did exhibit modest antibacterial activity on their own (Table 4 below). We evaluated the compounds described herein for their ability to lower the MIC of oxacillin against MRSA as described in Example 2 below.
The MIC of oxacillin against the resistant MRSA strain NRS70 is 32 μg/mL. Several inhibitors exhibited remarkable activity in lowering the MIC of oxacillin (Table 1) at the 20 μΜ level.
Embodiments of the Invention
The invention provides compounds of Formula A:
Figure imgf000022_0001
wherein
X is O, CH2, S, SO2, or a direct bond;
each R1 is independently CN, SO2CH3, CONH2, CO2H, C02(alkyl), CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or S02-(pyridinyl);
m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
each R2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R2 groups form an ortho-fused methylenedioxy; and
each R3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof, in addition to its sub-Formulas I, II, II-A, II-B, II-C, II-D, III, III-A, and III-B. When the groups R1 are different, a particular R1 can be E or Z with respect to the phenyl ring to which the alkene of the R1 is attached. Furthermore, when groups R2 and R3, are shown as attached at a variable location, they can be ortho, meta, or para to group X, or they can be excluded from locations ortho, meta, or para to group X. The invention also provides compounds of Formula I:
Figure imgf000023_0001
(I)
wherein
X is O, CH2, S, SO2, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO2H, CC Me, CO(aryl), pyridinyl, thiophi
benzothiazolyl, CO-cycloalkyl, or S02-(pyridinyl);
m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
each R2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, CC Me, NO2, OCF3, or SCF3, or two R2 groups form an ortho-fused methylenedioxy; and
each R3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof, in addition to its sub-Formulas II, II-A, II-B, II-C, II-D, III,
III-A, and III-B. The group R1 can be E or Z with respect to the phenyl ring to which the alkene of R1 is attached. In certain specific embodiments, the group R1 is in the Z configuration, and therefore the CN is in the Z configuration.
One specific value for X is O. Another specific value for X is CH2. Another specific value for X is S. Another specific value for X is SO2. Another specific value for X is a direct bond.
Specific values for R1 include CN, SO2CH3, CONH2, CO2H, and C02Me. Another specific value of R1 is CO(aryl), for example, CO(phenyl), wherein phenyl is optionally substituted with one or more substituents, such as methyl, halo, methoxy, amino, nitro, CN, CF3, and the like, or a substituent as described herein. Another specific value of R1 is CO(4-MePh).
When R1 is pyridyl, the pyridyl and be a 2-pyridyl, 3-pyridyl, or 4-pyridyl. When R1 is thiophenyl, the thiophenyl can be a 2- thiophenyl or a 3- thiophenyl. When R1 is benzothiazolyl, R1 can be 2- benzothiazolyl, 4- benzothiazolyl, 5- benzothiazolyl, 6- benzothiazolyl, or 7- benzothiazolyl.
Specific values of R1 when R1 is CO-cycloalkyl are CO-cyclopropyl, CO-cyclobutyl,
CO-cyclopentyl, or CO-cyclohexyl. Specific values of R1 when R1 is S02-(pyridinyl) are S02-(2-pyridinyl), S02-(3- pyridinyl), and SC -(4-pyridinyl).
In one embodiment, m can be 1. In another embodiment, m can be 2. In yet another embodiment, m can be 3. In a further embodiment, m can be 4.
In one embodiment, n can be 1. In another embodiment, n can be 2. In yet another embodiment, n can be 3. In a further embodiment, n can be 4. In yet a further embodiment, n can be 5.
Specific values for R2 include H, C≡CH, OH, C02H, CC Me, NO2, OCF3, and SCF3.
R2 can also be halo, for example, F, CI, Br, or I.
R2 can also be alkyl, for example, (Ci-C6)alkyl, optionally branched. Examples include methyl, ethyl, propyl, seopropyl, wo-propyl, seobutyl, fert-butyl, and the like.
R2 can also be alkoxy, for example, (C1-C6) alkoxy, optionally branched. Examples include methoxy, ethoxy, propoxy, seopropoxy, wo-propoxy, seobutoxy, fert-butoxy, and the like.
R2 can also be heteroaryl, for example, thiophenyl, furanyl,benzothiazolyl, pyridyl, or pyrimidinyl, substituted on the heteroaryl at any available valency.
R2 can also be heterocycle, for example, N-morpholinyl, tetrahydropyranyl,
tetrahydrofuranyl, or piperidyl, substituted on the heterocycle at any available valency.
In certain specific embodiments, two R2 groups form an ortho-fused methylenedioxy. Specific values for R3 include H, C≡CH, OH, CO2H, C02Me, ΝΟ2, OCF3, and SCF3.
R3 can also be halo, for example, F, CI, Br, or I.
R3 can also be alkyl, for example, (Ci-C6)alkyl, optionally branched. Examples include methyl, ethyl, propyl, seopropyl, wo-propyl, seobutyl, fert-butyl, and the like.
R3 can also be alkoxy, for example, (C1-C6) alkoxy, optionally branched. Examples include methoxy, ethoxy, propoxy, seopropoxy, wo-propoxy, seobutoxy, fert-butoxy, and the like.
R3 can also be heteroaryl, for example, thiophenyl, furanyl,benzothiazolyl, pyridyl, or pyrimidinyl, substituted on the heteroaryl at any available valency.
R3 can also be heterocycle, for example, N-morpholinyl, tetrahydropyranyl,
tetrahydrofuranyl, or piperidyl, substituted on the heterocycle at any available valency.
In certain specific embodiments, two R3 groups form an ortho-fused methylenedioxy.
In certain specific embodiments, a group or specific value of X, R1, R2, or R3 is excluded from various embodiments of the invention. The invention also provides the compounds illustrated in Figure 1. In some
embodiments, compounds 81, 82, 85-90, 93-96, and 98-99 are excluded from such
embodiments. General Synthetic Methods
In general, preparation of the compounds and formulas described herein, and modifications thereof, can be made according to organic synthesis techniques known to those of skill in the art and/or according to the synthetic schemes provided herein. Where desired, synthesis of a subject compound can begin with commercially available chemicals, from compounds described in the chemical literature, or from products of the reactions and methods described herein. Commercially available compounds may be obtained from standard commercial sources including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), ICN Biomedicals, Inc. (Costa Mesa, CA), Lancaster Synthesis (Windham, NH), Spectrum Quality Product, Inc. (New
Brunswick, NJ), TCI America (Portland, OR), Combi-Blocks, Inc. (San Diego, CA), Oakwood Products, Inc. (Estill, SC), and Wako Chemicals USA, Inc. (Richmond, VA).
In addition, methods known to one of ordinary skill in the art may be identified through various reference books and databases. Suitable reference books and treatises that detail the synthesis of reactants useful in the preparation of the inhibiting agents described herein, or provide references to articles that describe the preparation, include for example, "Synthetic Organic Chemistry", John Wiley & Sons, Inc., New York; S. R. Sandler et al, "Organic Functional Group Preparations," 2nd Ed., Academic Press, New York, 1983; H. O. House, "Modem Synthetic Reactions", 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L. Gilchrist, "Heterocyclic Chemistry", 2nd Ed., John Wiley & Sons, New York, 1992; J. March, "Advanced Organic Chemistry: Reactions, Mechanisms and Structure", 4th Ed., Wiley- Interscience, New York, 1992; and Protecting Groups in Organic Synthesis, Second Edition, Greene, T.W., and Wutz, P.G.M., John Wiley & Sons, New York.
Additional suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds described herein, or provide references to articles that describe the preparation, include for example, Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting Materials", Second, Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V. "Organic Chemistry, An Intermediate Text" (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R. C. "Comprehensive Organic Transformations: A Guide to Functional Group Preparations" 2nd Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Structure" 4th Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, T. W. G. "Organic Chemistry" 7th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; "Industrial Organic Chemicals: Starting Materials and
Intermediates: An Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; "Organic Reactions" (1942-2000) John Wiley & Sons, in over 55 volumes; and "Chemistry of Functional Groups" John Wiley & Sons, in 73 volumes.
Unless specified to the contrary, the reactions described herein take place at atmospheric pressure, generally within a temperature range from -10 °C to 200 °C. Further, except as otherwise specified, reaction times and conditions are intended to be approximate, e.g., taking place at about atmospheric pressure within a temperature range of about -10 °C to about 110 °C over a period of about 1 to about 24 hours; reactions left to run overnight average a period of about 16 hours.
A number of exemplary methods for preparation of the compounds of the invention are provided below. These methods are intended to illustrate the nature of such preparations are not intended to limit the scope of applicable methods. Other variations, such as adding various substituents (e.g., as defined above) on various alkyl, aryl, or heterocycle groups are included in the scope of the invention. Relevant starting materials can typically be purchased from the commercial suppliers cited above (e.g., from Sigma- Aldrich, Milwaukee, WI) or they can be prepared in a few standard steps from commercially available materials.
In certain embodiments, compounds of various formulas described herein can be prepared by the following representative methods, as illustrated by Schemes 1-3. Scheme 1
Figure imgf000026_0001
where X, m, n, R2, and R3 are as defined for Formula I above. Scheme 2.
Figure imgf000027_0001
where m, n, R2, and R3 are as defined for Formula I above. Scheme
Figure imgf000027_0002
where m, n, R2, and R3 are as defined for Formula I above. As would be readily recognized by one of skill in the art, the various R1 groups of Formulas A and I are installed by condensation of the appropriate disubstituted methylene group with the relevant aldehyde. The disubstituted methylene groups are commercially available or can be prepared by standard synthetic techniques known to those of skill in the art. For example, compound 35-b is prepared using NC-CH2-CO(cyclopropyl)and compound 36-b is prepared using NC-CH2-SC (2-pyridyl).
Combination Therapy
The compounds described herein may be administered alone or in combination with other therapeutic agents, such as antibiotic, anti-inflammatory or antiseptic agents such as antibacterial agents, anti-fungicides, anti-viral agents, and anti-parasitic agents. In some embodiments, a pharmaceutical composition comprises one or more compounds described herein and one or more antibiotic or antiseptic agents. Examples of suitable active agents include penicillins, cephalosporins, carbacephems, cephamycins, carbapenems, monobactams, aminoglycosides, glycopeptides, quinolones, tetracyclines, macrolides, and fluoroquinolones. Suitable antiseptic agents that can be used include iodine, silver, copper, chlorhexidine, polyhexanide and other biguanides, chitosan, acetic acid, and hydrogen peroxide. These agents may be incorporated as part of the same pharmaceutical composition or may be administered separately (concurrently or sequentially). The pharmaceutical compositions may also contain anti-inflammatory drugs such as steroids and macrolactam derivatives.
Several embodiments described herein relate to a pharmaceutical composition that includes one or more β-lactam antibiotics and one or more compounds described herein, β- Lactam antibiotics are bactericidal, and can act by inhibiting the synthesis of the peptidoglycan layer of bacterial cell walls. The peptidoglycan layer is important for cell wall structural integrity, especially in Gram-positive bacteria. Examples of β-lactam antibiotics include, but are not limited to, benzathine penicillin, benzylpenicillin (penicillin G), phenoxymethylpenicillin (penicillin V), procaine penicillin, methicillin, ceftadizim, ceftaroline, oxacillin, nafcillin, cloxacillin, dicloxacillin, flucloxacillin, temocillin, amoxicillin, ampicillin, co-amoxiclav, azlocillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, cephalosporins, cephalexin, cephalothin, cefazolin, cefaclor, cefuroxime, cefamandole, cephamycins, cefotetan, cefoxitin, ceftriaxone, cefotaxime, cefpodoxime, cefixime, ceftazidime, cefepime, cefpirome, imipenem, meropenem, ertapenem, faropenem, doripenem, monobactams, aztreonam, tigemonam, nocardicin A, and tabtoxinine-P-lactam. Administration of β-lactam antibiotics can be carried out in conjunction with administration of a compound described herein. The administration of the β-lactam antibiotic can be concurrent or sequential with respect to the administration of a compound described herein. For example, a compound described herein can be administered to a subject having a bacterial infection, and then an antibiotic can be administered, its activity potentiated by the administration of a compound described herein. The antibiotic can be a β- lactam antibiotic recited herein above. Furthermore, the antibiotic can be clavulanate, sulbactam, tazobactam, avibactam, MK-7655, or NXL105.
Some embodiments provide methods for inhibiting the growth and/or reproduction of susceptible organisms, and/or to increasing the sensitivity of susceptible organisms to β-lactam antibiotics. Susceptible organisms generally include gram positive and gram negative, aerobic and anaerobic organisms whose growth can be inhibited by embodiments described herein. Susceptible organisms include, but are not limited to, Staphylococcus, Lactobacillus,
Streptococcus, Streptococcus agalactiae, Sarcina, S. pneumoniae, S. pyogenes, S. mutans, Escherichia, Enter obacter , Klebsiella, Pseudomonas, Pseudomonas aeruginosa, Acinetobacter, Proteus, Campylobacter , Citrobacter, Nisseria, Bacillus anthracis, Bacillus cereus, Bacillus subtilis, Bacteroides, Peptococcus, Clostridium, Salmonella, Shigella, Serratia, Haemophilus, Brucella, Mycobacterium tuberculosis and similar organisms.
Pharmaceutical Formulations
The compounds described herein can be used to prepare therapeutic pharmaceutical compositions, for example, by combining the compounds with a pharmaceutically acceptable diluent, excipient, or carrier. The compounds may be added to a carrier in the form of a salt or solvate. For example, in cases where compounds are sufficiently basic or acidic to form stable nontoxic acid or base salts, administration of the compounds as salts may be appropriate.
Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids that form a physiologically acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, a-ketoglutarate, and β- glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, halide, sulfate, nitrate, bicarbonate, and carbonate salts.
Pharmaceutically acceptable salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid to provide a physiologically acceptable ionic compound. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example, calcium) salts of carboxylic acids can also be prepared by analogous methods.
The compounds of the formulas described herein can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms. The forms can be specifically adapted to a chosen route of administration, e.g., oral or parenteral administration, by intravenous, intramuscular, topical or subcutaneous routes.
The compounds described herein may be systemically administered in combination with a pharmaceutically acceptable vehicle, such as an inert diluent or an assimilable edible carrier. For oral administration, compounds can be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the food of a patient's diet. Compounds may also be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations typically contain at least 0.1% of active compound. The percentage of the compositions and preparations can vary and may conveniently be from about 0.5% to about 60%, about 1 % to about 25%, or about 2% to about 10%, of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions can be such that an effective dosage level can be obtained. The tablets, troches, pills, capsules, and the like may also contain one or more of the following: binders such as gum tragacanth, acacia, com starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as com starch, potato starch, alginic acid and the like; and a lubricant such as magnesium stearate. A sweetening agent such as sucrose, fructose, lactose or aspartame; or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring, may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propyl parabens as preservatives, a dye and flavoring such as cherry or orange flavor. Any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices.
The active compound may be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can be prepared in glycerol, liquid polyethylene glycols, triacetin, or mixtures thereof, or in a pharmaceutically acceptable oil. Under ordinary conditions of storage and use, preparations may contain a preservative to prevent the growth of microorganisms.
Pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions, dispersions, or sterile powders comprising the active ingredient adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. The ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and/or antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers, or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by agents delaying absorption, for example, aluminum monostearate and/or gelatin.
Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, optionally followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation can include vacuum drying and freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the solution.
For topical administration, compounds may be applied in pure form, e.g., when they are liquids. However, it will generally be desirable to administer the active agent to the skin as a composition or formulation, for example, in combination with a dermatologically acceptable carrier, which may be a solid, a liquid, a gel, or the like.
Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina, and the like. Useful liquid carriers include water, dimethyl sulfoxide (DMSO), alcohols, glycols, or water-alcohol/glycol blends, in which a compound can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants.
Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using a pump-type or aerosol sprayer.
Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses, or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.
Examples of dermatological compositions for delivering active agents to the skin are known to the art; for example, see U.S. Patent Nos. 4,992,478 (Geria), 4,820,508 (Wortzman), 4,608,392 (Jacquet et al), and 4,559,157 (Smith et al). Such dermatological compositions can be used in combinations with the compounds described herein where an ingredient of such compositions can optionally be replaced by a compound described herein, or a compound described herein can be added to the composition.
Useful dosages of the compounds described herein can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Patent No. 4,938,949 (Borch et al.). The amount of a compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient, and will be ultimately at the discretion of an attendant physician or clinician.
In general, a suitable dose will be in the range of from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg/kg/day, most preferably in the range of 15 to 60 mg/kg/day. In one embodiment, the invention provides a composition comprising a compound of the invention formulated in such a unit dosage form.
The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations.
The invention provides therapeutic methods of treating a bacterial infection in a mammal, which involve administering to a mammal having a bacterial infection an effective amount of a compound or composition described herein. A mammal includes a primate, human, rodent, canine, feline, bovine, ovine, equine, swine, caprine, bovine and the like. The ability of a compound of the invention to treat a bacterial infection may be determined by using assays well known to the art.
The invention also provides a kit comprising a packaging containing one or more doses of a first pharmaceutical formulation comprising a compound described herein or a
pharmaceutically acceptable salt thereof, and one or more doses of a second pharmaceutical formulation comprising an antibiotic, each together with written instructions directing the coadministration of the first pharmaceutical formulation and the second pharmaceutical formulation for the treatment of bacterial infection. In some embodiments, the first dose of the first pharmaceutical formulation comprises a loading dose of a compound described herein. In some embodiments, the first dose of the second pharmaceutical formulation comprises a loading dose of an antibiotic. The individual doses of the pharmaceutical formulations, can
independently be in any dosage form, e.g. tablets, capsules, solutions, creams, etc. and packaged within any of the standard types of pharmaceutical packaging materials, e.g. bottles, blister- packs, IV bags, syringes, etc., that may themselves be contained within an outer packaging material such as a paper/cardboard box. In some embodiments, the kit further comprises one or more of culture media, culture plates, PCR primers, test strips, and/or stains for identifying the infective agent. The following Examples are intended to illustrate the above invention and should not be construed as to narrow its scope. One skilled in the art will readily recognize that the Examples suggest many other ways in which the invention could be practiced. It should be understood that numerous variations and modifications may be made while remaining within the scope of the invention.
EXAMPLES
Example 1. Preparation of Phosphorylation Inhibitors
General information. Reagents for chemical synthesis were purchased from Sigma-
Aldrich Chemical Co. (St. Louis, MO, U.S.A.), Combi -Blocks, Inc. (San Diego, CA, U.S.A.), Oakwood Products, Inc. (Estill, SC, U.S.A) or Alfa Aesar (Ward Hill, MA, U.S.A.). ¾ and 13C NMR spectra were acquired on a Varian DirectDrive 600 or a Varian INOVA-500 NMR spectrometer. High-resolution mass spectra were acquired on a Bruker microTOF/Q2 mass spectrometer (Bruker Daltonik, Bremen, Germany) by electrospray ionization. Thin-layer chromatography was done on EMD Millipore (Billerica, MA, U.S.A.) 0.25 mm silica gel 60 F254 plates. Column chromatography was done either manually using silica gel 60, 230-400 mesh (40-63 μιτι particle size) purchased from Sigma- Aldrich Chemical Co., or on a Teledyne Combiflash Rf 200i automated chromatography system (Teledyne Isco, Lincoln, NE, U.S.A.) using disposable silica gel columns.
Preparation of Compounds 1-11 :
2-(3-(3,4-d
Figure imgf000033_0001
To oven dried 25 ml round bottom flask, under a nitrogen atmosphere, was added 3-(3,4- dichlorophenoxy)benzaldehyde (160 mg, 0.60 mmol), dry ethanol (5 mL), malononitrile (40 mg, 0.60 mmol) and L-Proline (35 mg, 0.30 mmol). The reaction was stirred overnight and the precipitated was filtered and washed with hexane affording a white solid 92% yield (170 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 7.10 (dd, J=10.22, 2.75 Hz, 1H) 7.41 (dd, J=2.36, 0.79 Hz, 1H) 7.43 (d, J=2.75 Hz, 1H) 7.52 - 7.60 (m, 1H) 7.60 - 7.70 (m, 2H) 7.70 - 7.78 (m, 1H) 8.52 (s, 1H). 13C NMR (101 MHz, DMSO-de) δ ppm 83.52, 113.64, 114.64, 120.24, 120.41, 121.97, 125.04, 126.62, 127.17, 132.17, 132.48, 133.00, 133.75, 155.84, 157.22, 161.26. HRMS (m/z): [M - H]\ calcd for C16H7CI2N2O, 312.9941; found, 312.9951.
-(3-(3,4-dichlorobenzyl)benzylidene)malononitrile (2).
Figure imgf000034_0001
3-(3,4-dichlorobenzyl)benzaldehyde:
Under argon atmosphere to a mixture of THF (25 mL) and water (10 mL) were added potassium carbonate (2.76 g, 20 mmol), 3-formylphenylboronic acid (1.00 g, 6.67 mmol), 3,4- dichlorobenzylbromide (1.45 g, 6.06 mmol), and Pd(PPh3)4 (210 mg, 0.18 mmol). The reaction was heated to 80 °C overnight and the morning after quenched with aqueous HCl (1 M), and the aqueous phase was extracted with ethyl acetate. The combined organic layers were dried using MgS04, and solvent was removed in vacuo giving the crude product. The crude was purified with a gradient column chromatography from EtOAc/Hexane 1:9 affording a colorless liquid in 68% yield (1.09 g). ¾ NMR (500 MHz, CDCb-fife) δ ppm 7.02 (dd, J=8.56, 2.20 Hz, 1 H) 7.26 (d, J=3.18 Hz, 1 H) 7.36 (d, J=8.07 Hz, 1 H) 7.43 (dd, J=7.58, 0.49 Hz, 1 H) 7.48 (t, J=7.58 Hz, 1 H) 7.68 (s, 1 H) 7.75 (d, J=7.58 Hz, 1 H) 9.99 (s, 1 H). 13C NMR (101 MHz, CDCh-de) δ ppm 69.65, 112.28, 114.98, 115.60, 117.07, 117.77, 118.74, 127.81, 131.03, 131.12, 132.74, 135.35, 141.73, 157.42. HRMS (m/z): [M - H]\ calcd for C14H10CI2O, 265.0181; found, 265.0196.
2-(3-(3,4-dichlorobenzyl)benzylidene)malononitrile (2) :
To oven dried 25 ml round bottom flask, under a nitrogen atmosphere, was added 3-(3,4- dichlorobenzyl)benzaldehyde (123 mg, 0.46 mmol), dry ethanol (4 mL), malononitrile (30 mg, 0.46 mmol) and L-Proline (26 mg, 0.23 mmol). The reaction was stirred overnight and the precipitated was filtered and washed with hexane obtaining a white solid in 90% yield (131 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 4.05 (s, 2 H) 7.20 - 7.27 (m, 1 H) 7.59 (m, J=8.10 Hz, 4 H) 7.73 - 7.79 (m, 1 H) 7.82 (dt, J=7.40, 1.44 Hz, 1 H) 8.51 (s, 1 H). 13C NMR (101 MHz, DMSO-de) 5 ppm 39.77, 82.27, 113.66, 114.64, 128.69, 129.49, 129.70, 130.29, 131.20, 131.24, 131.57, 132.10, 135.22, 142.01, 142.27, 161.94. HRMS (m/z): [M - H]\ calcd for C17H10N2CI2, 311.0148; found, 311.0185.
Figure imgf000035_0001
2-(5-(3,4-dichlorobenzyl)-2-fluorobenzylidene)malononitrile (3).
This compound was prepared according to the procedure for 2 in 62% yield (135 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 4.02 (s, 2 H) 7.24 (dd, J=8.19, 1.83 Hz, 1 H) 7.40 (dd, J=10.03, 8.80 Hz, 1 H) 7.50 - 7.59 (m, 2 H) 7.60 - 7.70 (m, 1 H) 7.88 (dd, J=6.72, 1.59 Hz, 1 H) 8.55 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 39.30, 85.78, 113.31, 114.38, 117.34, 117.51, 120.10, 120.20, 129.74, 129.81, 131.41, 131.84, 137.33, 137.40, 138.24, 138.27, 142.15, 154.74, 154.77, 158.62, 160.65. HRMS (m/z): [M - H]\ calcd for C17H9CI2FN2, 329.0054;
found, 329.0088.
Figure imgf000035_0002
2-(3-(3,4-dichlorobenzyl)-2-fluorobenzylidene)malononitrile (4).
This compound was prepared according to the procedure for 2 in 51% yield (110 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 4.07 (s, 2 H) 7.23 (dd, J=8.19, 2.08 Hz, 1 H) 7.42 (t, J=7.70 Hz, 1 H) 7.55 (d, J=1.96 Hz, 1 H) 7.58 (d, J=8.31 Hz, 1 H) 7.72 (td, J=7.60, 1.70 Hz, 1 H) 7.89 - 8.02 (m, 1 H) 8.60 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 33.33, 85.48, 113.21, 114.23, 120.11, 120.23, 125.81, 125.85, 128.05, 128.74, 128.90, 129.50, 129.64, 131.06, 131.22, 131.54, 137.68, 137.74, 140.78, 154.46, 154.53, 157.60, 160.16. HRMS (m/z): [M - H]\ calcd for C17H9CI2FN2, 329.0054; found, 329.0029.
Figure imgf000035_0003
2-(5-(3,4-dichlorobenzyl)-2-methoxybenzylidene)malononitrile (5). This compound was prepared according to the procedure for 2 in 61% yield (90 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.88 (s, 3 H) 3.95 (s, 2 H) 7.18 (d, J=8.56 Hz, 1 H) 7.22 (dd, J=8.19, 2.08 Hz, 1 H) 7.52 (d, J=1.96 Hz, 1 H) 7.54 (d, J=8.07 Hz, 1 H) 7.58 (dd, J=8.56, 2.20 Hz, 1 H) 7.80 (d, J=2.20 Hz, 1 H) 8.43 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 39.07, 56.64, 82.26, 113.19, 113.68, 114.86, 120.40, 129.30, 129.49, 131.04, 131.13, 131.53, 133.32, 137.25, 142.54, 156.56, 157.11. HRMS (m/z): [M - H]\ calcd for C18H12CI2N2O, 341.0253; found, 341.0246.
Figure imgf000036_0001
(E)-3-(5-(3,4-dichlorobenzyl)-2-fluorophenyl)-2-(methylsulfonyl)acrylonitrile (6). This compound was prepared according to the procedure for 2 in 57% yield (127 mg). ¾
NMR (400 MHz, DMSO-c e) δ ppm 3.42 (s, 3H) 4.04 (s, 2H) 7.26 (dd, J=8.26, 1.97 Hz, 1H) 7.43 (dd, J=10.23, 8.65 Hz, 1H) 7.50 - 7.61 (m, 2H) 7.66 (ddd, J=8.06, 5.31, 1.97 Hz, 1H) 7.97 (dd, J=6.69, 1.57 Hz, 1H) 8.26 (s, 1H). 13C NMR (101 MHz, DMSO-de) δ ppm 39.34, 42.52, 113.51, 116.52, 117.37, 117.58, 118.96, 119.08, 129.72, 129.85, 131.44, 131.87, 137.28, 137.38, 138.37, 138.41, 142.21, 146.04, 146.10, 158.99, 161.52. HRMS (m/z): [M - H]+, calcd for C17H13CI2FNO2S, 384.0023; found, 383.996.
Figure imgf000036_0002
(E)-3-(3-(3,4-dichlorobenzyl)-2-fluorophenyl)-2-(methylsulfonyl)acrylonitrile (7).
This compound was prepared according to the procedure for 2 in 53% yield (120 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.43 (s, 3 H) 4.09 (s, 2 H) 7.25 (dd, J=8.19, 2.08 Hz, 1 H) 7.45 (t, J=7.70 Hz, 1 H) 7.56 (d, J=1.96 Hz, 1 H) 7.58 (d, J=8.31 Hz, 1 H) 7.74 (td, J=7.46, 1.47 Hz, 1 H) 8.06 (dt, J=8.07, 1.96 Hz, 1 H) 8.31 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 33.36, 42.34, 113.36, 116.12, 116.15, 118.95, 119.07, 125.93, 125.98, 127.95, 128.80, 128.96, 129.50, 129.64, 131.04, 131.20, 131.55, 137.65, 137.71, 140.80, 145.89, 145.97, 158.25, 160.81. HRMS (m/z): [M - H]\ calcd for C17H12CI2FNO2S, 381.9877; found, 381.9855.
Figure imgf000037_0001
(E)-3-(3-(3,4-dichlorobenzyl)-2,6-difluorophenyl)-2-(methylsulfonyl)acrylonitrile
(8).
This compound was prepared according to the procedure for 2 in 35% yield (62 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.47 (s, 3 H) 4.06 (s, 2 H) 7.25 (dd, J=8.19, 2.08 Hz, 1 H) 7.36 (t, 3=9.17 Hz, 1 H) 7.55 (d, J=1.96 Hz, 1 H) 7.58 (d, J=8.31 Hz, 1 H) 7.75 (td, J=8.68, 6.60 Hz, 1 H) 8.21 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 32.93, 32.96, 42.10, 109.13, 109.31, 109.48, 112.35, 112.91 , 1 12.95, 1 13.12, 1 13.16, 121.26, 124.64, 124.68, 124.80, 124.84, 129.46, 129.66, 131.01, 131.21 , 131.57, 136.85, 136.93, 136.96, 137.03, 140.73, 142.45, 156.72, 156.77, 156.82, 157.30, 157.67, 157.72, 159.31 , 159.37, 160.20, 160.26. HRMS (m/z): [M - H]\ calcd for C17H11CI2F2NO2S, 399.9783; found, 399.9743.
Figure imgf000037_0002
(E)-3-(5-(3,4-dichlorobenzyl)-2-methoxyphenyl)-2-(methylsulfonyl)acrylonitrile (9).
This compound was prepared according to the procedure for 2 in 72% yield (103 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.37 (s, 3 H) 3.90 (s, 3 H) 3.97 (s, 2 H) 7.21 (d, J=8.80 Hz, 1 H) 7.23 (dd, J=8.10, 1.96 Hz, 1 H) 7.54 (s, 1 H) 7.54 (d, J=10.76 Hz, 1 H) 7.60 (dd, J=8.56, 2.20 Hz, 1 H) 7.92 (d, J=1.96 Hz, 1 H) 8.40 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 39.09, 42.37, 56.72, 1 13.19, 113.27, 113.96, 118.90, 129.04, 129.34, 129.53, 131.09, 131.16, 131.55, 133.43, 137.22, 142.55, 147.65, 158.20. HRMS (m/z): [M + H]+, calcd for
C18H15CI2NO3S, 396.0222; found,
Figure imgf000037_0003
(E)-3-(3-(3,4-dichlorobenzyl)-2-methoxyphenyl)-2-(methylsulfonyl)acrylonitrile
(10).
This compound was prepared according to the procedure for 2 in 70% yield (148 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.42 (s, 3 H) 3.75 (s, 3 H) 4.05 (s, 2 H) 7.22 (dd, J=8.31, 1.96 Hz, 1 H) 7.38 (t, J=7.83 Hz, 1 H) 7.52 (d, J=2.20 Hz, 1 H) 7.56 (d, J=8.31 Hz, 1 H) 7.60 (dd, J=7.58, 1.47 Hz, 1 H) 8.03 (dd, J=7.95, 1.34 Hz, 1 H) 8.35 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 34.45, 42.43, 64.07, 113.64, 114.87, 124.50, 125.73, 127.78, 129.35, 129.59, 131.06, 131.42, 135.15, 137.49, 141.83, 148.49, 159.29. HRMS (m/z): [M + H]+, calcd for C18H15CI2NO3S, 396.0222; found, 396.0199.
Figure imgf000038_0001
(E)-3-(5-(3,4-dichlorobenzyl)-2-(trifluoromethoxy)phenyl)-2- (methylsulfonyl)acrylonitrile (11).
This compound was prepared according to the procedure for 2 in 63% yield (102 mg). ¾ NMR (400 MHz, DMSO-ife) δ ppm 3.41 (s, 3 H) 4.08 (s, 2 H) 7.27 (d, J=7.87 Hz, 1 H) 7.44 - 7.65 (m, 3 H) 7.71 (d, J=7.87 Hz, 1 H) 8.04 (s, 1 H) 8.29 (s, 1 H). 13C NMR (101 MHz, DMSO- de) 5 ppm 38.69, 41.76, 112.41, 114.96, 117.38, 118.54, 121.11, 121.85, 123.43, 123.68, 129.14, 129.39, 130.70, 130.74, 131.17, 135.60, 140.66, 141.02, 145.50, 145.51, 146.20. HRMS (m/z): [M - H]\ calcd for C18H12CI2F3NO3S, 447.9794; found, 447.9765. Preparation of Compound 73:
(E)-3-(3',4'-dichloro-4-fluoro-[l,l'-biphenyl]-3-yl)-2-(methylsulfonyl)acrylonitrile (73).
Figure imgf000038_0002
3',4'-dichloro-4-fluoro-[l, -biphenyl]-3-carbaldehyde:
Under argon atmosphere to a mixture of THF (25 mL) and water (10 mL) were added potassium carbonate (2.76 g, 20 mmol), 2-fluoro-3-formylphenylboronic acid (1.00 g, 5.95 mmol), 3,4-dichlorobromobenzene (1.22 g, 5.41 mmol), and Pd(PPh3)4 (188 mg, 0.16 mmol). The reaction was heated to 80 °C overnight and the morning after quenched with aqueous HC1 (1 M), and the aqueous phase was extracted with ethyl acetate. The combined organic layers were dried using MgS04, and solvent was removed in vacuo giving the crude product. The crude was purified with a gradient column chromatography from EtOAc/Hexane 1 :9 affording a white solid in 55% yield (0.80 g). ¾ NMR (400 MHz, DMSO-c e) δ ppm 7.49 (dd, J=10.39, 8.68 Hz, 1 H) 7.62 - 7.74 (m, 2 H) 7.96 (dd, J=1.71, 0.73 Hz, 1 H) 8.02 - 8.15 (m, 2 H) 10.23 (s, 1 H). 13C NMR (101 MHz, DMSO-de) δ ppm 117.92, 118.12, 124.49, 124.58, 127.40, 128.61, 129.04, 131.27, 131.52, 132.33, 134.86, 134.89, 135.32, 135.41, 138.97, 162.13, 164.71, 188.30, 188.34. HRMS (m/z): [M + H]+, calcd for C13H7C12FO, 268.9931; found, 268.9936.
(E)-3-(3^4'-dichloro-4-fluoro-[l,r-biphenyl]-3-yl)-2-(methylsulfonyl)acrylonitrile (73):
To oven dried 25 ml round bottom flask, under a nitrogen atmosphere, was added 3 ',4'- dichloro-4-fluoro-[l,l'-biphenyl]-3-carbaldehyde (135 mg, 0.50 mmol), dry ethanol (4 mL), (methylsulfonyl)acetonitrile (60 mg, 0.50 mmol) and L-proline (29 mg, 0.25 mmol). The reaction was stirred overnight and the precipitated was filtered and washed with hexane obtaining a white solid in 90% yield (167 mg). ¾ NMR (400 MHz, DMSO-c e) δ ppm 3.47 (s, 3 H) 7.60 (t, J=9.54 Hz, 1 H) 7.69 (dd, J=8.44, 2.08 Hz, 1 H) 7.79 (d, J=8.56 Hz, 1 H) 7.98 (d, J=1.96 Hz, 1 H) 8.05 - 8.16 (m, 1 H) 8.37 (s, 1 H) 8.43 (dd, J=6.85, 2.20 Hz, 1 H). 13C NMR (101 MHz, DMSO-de) 5 ppm 42.44, 113.39, 117.49, 117.69, 117.90, 119.37, 119.50, 127.29, 128.59, 129.04, 131.50, 131.77, 132.46, 134.83, 134.93, 135.02, 135.06, 138.96, 146.26, 146.30, 159.78, 162.34.
Example 2. Compound MIC Data
Minimal-inhibitory concentration (MIC) determination. Determination of MICs was carried out by the microdilution method cation-adjusted in Mueller-Hinton II Broth (CAMHB II, BBL) in accordance with the protocols of CLSI {Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Second Informational Supplement. CLSI document M100-S22. Clinical and Laboratory Standards Institute. Wayne, PA). A final bacterial inoculum of 5 χ 105 CFU/rnL was used, and the results were recorded after incubation for 16-20 h at 37 °C.
The MIC values of 100 compounds were evaluated. Numerous compounds were evaluated at three or more different concentrations, typically: 0 (control), 2, and 20 μΜ. If the compound exhibited inherent antibacterial activity at 20 μΜ against the S. aureus strain
(NRS70), the compound was reevaluated at lower concentrations for the particular strain (e.g., 0, 0.1, and 1 μΜ, or 0, 5, and 10 μΜ) to eliminate interference with the assay. Several compounds showed a greater than 2-fold decrease relative to the control.
Table 1. Minimal-inhibitory concentration (MIC) for oxacillin (OXA) in the presence of 20 μΜ of potentiator (compounds 1-52). The symbol asterisk indicates no growth under the defined condition.
Figure imgf000040_0001
Figure imgf000040_0002
22-a Ri = CN, R3 = 3-Cl, 5-Cl 0.06
23 -a Ri = CN, R3 = 4-OMe
24-a Ri = CO(4-MePh), R3 = 3-Cl, 4-Cl ; i ¾
25 -a Ri = 2-Thiophene, R3 = 3-Cl, 4-Cl
26-a Ri = 2-Benzothiazole, R3 = 3-Cl, 4-Cl
27-a Ri = 4-Pyridine, R3 = 3-Cl, 4-Cl 4
28-a Ri = 2-Pyridine, R3 = 3-Cl, 4-Cl
29-a Ri = CN, R2 = 6-OMe, R3 = 3-Cl, 4-Cl 4
30-b Ri = CN R3 = 2-F, 4-F, 5-F
31-b Ri = CN, R3 = 3-F, 4-Cl
32-b Ri = CN, R3 = 3-Cl, 4-F - _ <i
33-b Ri = CN, R3 = 2-OMe, 5-OMe
34-b Ri = CN, R3 = 3-Cl
35-b Ri = CO-Cyclopropyl, R2 = 4-F, R3 = 3-Cl, 4-Cl
36-b Ri = S02-(2-Pyridinyl), R2 = 4-F, R3 = 3-Cl, 4-Cl (; 00*
37-b Ri = S02CH3, R2 = 4-F, R3 = 4-CF3
38-b Ri = S02CH3, R2 = 4-F, R3 = 3-CF3 16
39-b Ri = S02CH3, R2 = 4-F, R3 = 3-F, 4-F
40-b Ri = S02CH3, R2 = 4-F, R3 = 3-CF3, 4-OMe 0 '? 'S
41 -b Ri = S02CH3, R2 = 4-F, R3 = 3-CF3, 4-F 4
42-b Ri = S02CH3, R2 = 4-F, R3 = 3-F, 4-CF3 0. ό
43 -b Ri = S02CH3, R2 = 4-F, R3 = 4-OCF3 4
44-b Ri = S02CH3, R2 = 4-F, R3 = 3-OCF3 1 6
45 -b Ri = S02CH3, R2 = 4-F, R3 = 4-CONHCH(CH3)2
46-b Ri = S02CH3, R2 = 4-F, R3 = 4-SCF3 8
47-b Ri = S02CH3, R2 = 4-OEt, R3 = 3-Cl, 4-Cl
48-b Ri = S02CH3, R2 = 2-OMe, 4-OMe, R3 = 3-Cl, 4-Cl 16
49-b Ri = S02CH3, R2 = 4-OCF3, R3 = 3-OMe, 4-OMe 0 1 '7
50-b Ri = S02CH3, R2 = 4-OCF3, R3 = 3,4-Methylenedioxy
51-b Ri = S02CH3, R2 = 4-N-Morpholine, R3 = 3-Cl, 4-Cl v.!. Vf
52-b Ri = S02CH3, R2 = 4-OH, R3 = 3-Cl, 4-Cl 16
ESP388 Ri = S02CH3, C02Et (in place of CN; i.e., a compound
of Formula A), R2 = 2-F, 4-F, R3 = 3-Cl, 4-Cl Table 2. Additional structures of the same class that do not conform to the general template structure shown at the top of Table 1. MIC for oxacillin in the presence of 20 μΜ of potentiator.
Figure imgf000042_0001
Compounds 60-62:
Figure imgf000042_0002
Figure imgf000043_0001
Table 3. Additional compounds of a distinct molecular template. MIC for oxacillin i presence of 20 μΜ of potentiator.
Figure imgf000043_0002
Figure imgf000044_0001
Table 4. Individual potentiator compounds shown with their molecular structures, along with the MIC for the compound against strain NRS70 and MIC of oxacillin in the presence of 20 μΜ of the potentiator against NRS70. The symbol asterisk indicates no growth under the defined condition.
Figure imgf000045_0001
40 >256 0.25
41 >256 4
42 32 0.06
43 64 4
44 64 16
45 >256 >32
46 >256 8
47 >256 0.06
48 16 16
49 64 0.12
50 32 0.06
51 >256 0.06
52 16 16
53 >256 >32
54 64 >32
55 64 16
56 128 >32
57 >256 >32
58 >256 >32
59 64 >32
125
60 32 16
61 64 >32
62 >256 >32
63 >256 >32
64 >256 >32
65 >256 >32
66 >256 >32
67 64 >32
68 >256 >32
69 >256 >32
70 >256 >32
71 >256 >32
72 >256 >32
73 16 0.03
74 32 0.03
75 >256 >32
76 >256 >32
77 >256 >32
78 >256 >32
79 32 4
80 32 16
81 32 >32
82 32 16
83 64 >32
84 >256 >32
85 32 >32 86 >256 >32
87 >256 >32
88 >256 >32
89 >256 >32
90 >256 >32
91 >256 >32
92 >256 >32
93 >256 >32
94 >256 >32
95 >256 >32
96 >256 >32
97 >256 >32
98 >256 >32
99 >256 >32
Table 5. Potentiators 1 to 11 and ESP388 were evaluated in greater depth. The compounds at 20, 16, 12 and 8 μΜ were evaluated as potentiators and as antibiotics. MRSA activity was tested against 7 different MRSA strains: NRS70, MRSA252 (USA200), NRS 123 (USA400), NRS382 (USA100), NRS384 (USA300), NRS1 (Vancomycin-Intermediate Staphylococcus aureus - VISA) and NRS 119 (a linezolid-Resistant Staphylococcus aureus).
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000048_0002
Compound number and MIC Oxacillin MIC with 20 molecular structure ug/mL μΜ of compound
4 MRSA252 20 μΜ = 6.6 μ /ml
8 MRSA252 NRS70
NRS1 0.03 0.03
8 NRS123 MRSA119
NRS70 0.06 0.015
8 NRS1 NRS382
CI NRS123
0.03 0.12 8 NRS384
NRS119
0.03
8
NRS382
8
NRS384
8
Figure imgf000049_0002
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000050_0002
Figure imgf000050_0004
Figure imgf000050_0003
Figure imgf000050_0005
>252
NRS382
>252
NRS384
>252
Figure imgf000051_0001
Figure imgf000051_0002
Compound number and MIC ug/mL Oxacillin MIC with 16 Oxacillin MIC with 12 molecular structure μΜ of compound μΜ of compound
ESP388 MRSA252 16 μΜ = 7.2 vg/m L 12 μΜ = 5.4 μ /ml
8 MRSA252 NRS70 MRSA252 NRS70
NRS1 64 0.06 128 1
1 c 8 NRS123 MRSA119 NRS123 MRSA119
NRS70 0.12 0.03 0.50 0.25 8 NRS1 NRS382 NRS1 NRS382
NRS123
0.03 0.12 4 0.25 8 NRS384 NRS384
CI NRS119
0.12 1
16
NRS382
16
NRS384
16
Table 6. Potentiation activity of compounds 7 and 8 with two additional antibiotics, meropenem (MER) and ceftazidime (CEF). We also calculated the MBC (minimal bactericidal concentration).
Figure imgf000052_0001
Figure imgf000053_0001
Example 3. Pharmaceutical Dosage Forms
The following formulations illustrate representative pharmaceutical dosage forms that may be used for the therapeutic or prophylactic administration of a compound of a formula described herein, a compound specifically disclosed herein, or a pharmaceutically acceptable salt or solvate thereof (hereinafter referred to as 'Compound X'):
(!) Tablet 1 mg/tablet
'Compound X' 100.0
Lactose 77.5
Povidone 15.0
Croscarmellose sodium 12.0
Microcrystalline cellulose 92.5
Magnesium stearate 3.0
300.0
(ii) Tablet 2 mg/tablet
'Compound X' 20.0
Microcrystalline cellulose 410.0
Starch 50.0
Sodium starch glycolate 15.0
Magnesium stearate 5.0
500.0
(iii) Capsule mg/capsule
'Compound X' 10.0
Colloidal silicon dioxide 1.5
Lactose 465.5
Pregelatinized starch 120.0 Magnesium stearate 3.0
600.0
(iv) Injection 1 (1 mg/mL) mg/mL 'Compound X' (free acid form) 1.0
Dibasic sodium phosphate 12.0
Monobasic sodium phosphate 0.7
Sodium chloride 4.5 1.0 N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5)
Water for injection q.s. ad 1 mL
(v) Injection 2 (10 mg/mL) mg/mL
'Compound X' (free acid form) 10.0 Monobasic sodium phosphate 0.3 Dibasic sodium phosphate 1.1
Polyethylene glycol 400 200.0 0.1 N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5)
Water for injection q.s. ad 1 mL
(vi) Aerosol mg/can
'Compound X' 20 Oleic acid 10 Trichloromonofluoromethane 5,000 Dichlorodifluoromethane 10,000 Dichlorotetrafluoroethane 5,000
(vii) Topical Gel 1 wt.%
'Compound X 5%
Carbomer 934 1.25%
Triethanolamine q.s.
(pH adjustment to 5-7)
Methyl paraben 0.2% Purified water q.s. to lOOg
(viii) Topical Gel 2 wt.%
'Compound X 5%
Methylcellulose 2%
Methyl paraben 0.2%
Propyl paraben 0.02%
Purified water q.s. to lOOg
(ix) Topical Ointment wt.%
'Compound X 5%
Propylene glycol 1%
Anhydrous ointment base 40%
Polysorbate 80 2% Methyl paraben 0.2%
Purified water q.s. to lOOg
(x) Topical Cream 1 wt.%
'Compound X' 5%
White bees wax 10%
Liquid paraffin 30%
Benzyl alcohol 5%
Purified water q.s. to lOOg
(xi) Topical Cream 2 wt.%
'Compound X' 5%
Stearic acid 10%
Glyceryl monostearate 3%
Polyoxy ethylene stearyl 3%
Sorbitol 5%
Isopropyl palmitate 2 %
Methyl Paraben 0.2%
Purified water q.s. to lOOg
These formulations may be prepared by conventional procedures well known in the pharmaceutical art. It will be appreciated that the above pharmaceutical compositions may be varied according to well-known pharmaceutical techniques to accommodate differing amounts and types of active ingredient 'Compound X'. Aerosol formulation (vi) may be used in conjunction with a standard, metered dose aerosol dispenser. Additionally, the specific ingredients and proportions are for illustrative purposes. Ingredients may be exchanged for suitable equivalents and proportions may be varied, according to the desired properties of the dosage form of interest. While specific embodiments have been described above with reference to the disclosed embodiments and examples, such embodiments are only illustrative and do not limit the scope of the invention. Changes and modifications can be made in accordance with ordinary skill in the art without departing from the invention in its broader aspects as defined in the following claims.
All publications, patents, and patent documents are incorporated by reference herein, as though individually incorporated by reference. No limitations inconsistent with this disclosure are to be understood therefrom. The invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

Claims

What is claimed is:
1. A compound of Formula I:
Figure imgf000056_0001
wherein
X is O, CH2, S, SO2, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO2H, CC Me, CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or SC -(pyridinyl);
m is 1 , 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
each R2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R2 groups form an ortho-fused methylenedioxy; and
each R3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
2. The compound of claim 1 wherein X is O.
3. The compound of claim 1 wherein X is CH2.
4. The compound of claim 1 wherein X is S or a direct bond.
5. The compound of any one of claims 1 -4 wherein R1 is CN or SO2CH3.
6. The compound of claim 1 wherein R2 is H, F, OMe, OEt, OCF3, SCF3, N- morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy.
7. The compound of claim 1 wherein R3 is H, F, OMe, OEt, OCF3, SCF3, N- morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy.
8. The compound of claim 1 wherein X is O, CH2, S, or a direct bond, R1 is CN or SO2CH3, m is 1 or 2, and n is 1 or 2.
9. The compound of claim 8 wherein R2 is H, F, OMe, OEt, OCF3, SCF3, N- morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy; and R3 is CI, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy.
10. The compound of claim 1 wherein the compound is a compound of Formula II:
Figure imgf000057_0001
(II)
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or SO2- (pyridinyl);
m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
R2 is H, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, N-morpholinyl, or pyridinyl, or two R3 groups form an ortho-fused methylenedioxy;
or a salt or solvate thereof.
11. The compound of cl d is a compound of Formula II-A:
Figure imgf000057_0002
(Π-Α)
wherein
is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or SO2-
(pyridinyl);
m is 1 or 2; n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy;
or a salt or solvate thereof.
12. The compound of cl d is a compound of Formula II-B:
Figure imgf000058_0001
(II-B)
wherein
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or SO2- (pyridinyl);
m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy;
or a salt or solvate thereof.
13. The compound of cl d is a compound of Formula II-C:
Figure imgf000058_0002
(II-C)
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or SO2- (pyridinyl);
m is 1 or 2;
n is 1 or 2; R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy;
or a salt or solvate thereof.
14. The compound of cl d is a compound of Formula II-D:
Figure imgf000059_0001
(II-D)
wherein
R1 is CN, SO2CH3, CONH2, CO(MePh), pyridinyl, CO-cyclopropyl, or SO2- (pyridinyl);
m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, or N-morpholinyl, or two R2 groups form an ortho-fused methylenedioxy; and
R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy;
or a salt or solvate thereof.
15. The compound of claim 1 wherein the compound is a compound of Formula III:
Figure imgf000059_0002
(III)
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S02-(2- pyridinyl);
m is 1 or 2;
n is 1 or 2;
R2 is H, F, OMe, OEt, OCF3, SCF3, or N-morpholinyl; and R3 is CI, F, OMe, OEt, OCF3, SCF3, or pyridinyl, or two R3 groups form an ortho- fused methylenedioxy;
or a salt or solvate thereof.
16. The compound of cl d is a compound of Formula III-A:
Figure imgf000060_0001
(III-A)
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(4-MePh), 2-pyridinyl, CO-cyclopropyl, or S02-(2- pyridinyl);
m is 1 or 2; and
R2 is H, F, OMe, OEt, OCF3, SCF3, or N-morpholinyl;
or a salt or solvate thereof.
17. The compound of cl und is a compound of Formula III-B:
Figure imgf000060_0002
(III-B)
wherein
X is O, CH2, S, or a direct bond;
R1 is CN, SO2CH3, CONH2, CO(aryl), pyridinyl, CO-cyclopropyl, or S02-(pyridinyl); n is 2;
R2 is F; and
R3 is CI;
or a salt or solvate thereof.
18. A compound illustrated in Figure 3, or a salt or solvate thereof.
19. A composition comprising a compound of any one of claims 1-18 in combination with a pharmaceutically acceptable diluent, excipient, or carrier.
20. A composition comprising a compound of any one of claims 1-18 in combination with a β-lactam antibiotic.
21. The composition of claim 20 wherein the β-lactam antibiotic is ceftadizim, ceftaroline, ceftazidime, meropenem, oxacillin, or penicillin.
22. A method to reverse the methicillin-resistant phenotype in a methicillin-resistant bacterium comprising contacting methicillin-resistant Staphylococcus aureus (MRSA) with an effective amount of a compound of any one of claims 1-17, thereby rendering the MRSA susceptible to β-lactam antibiotics.
23. A method to inhibit or kill methicillin-resistant Staphylococcus aureus (MRSA) comprising contacting the MRSA with an amount of a compound of any one of claims 1-17 effective to reverse the methicillin-resistant phenotype, and contacting the MRSA with an effective antibacterial amount of a β-lactam antibiotic.
24. A method to attenuate or reduce the minimum inhibitory concentration (MIC) of a β- lactam antibiotic comprising contacting a bacterium with an effective amount of a compound of any one of claims 1-17, in combination with contacting the bacterium with a β-lactam antibiotic.
25. A method to treat a patient infected with a bacterium resistant to a β-lactam antibiotic comprising administering to the patient an effective amount of a compound of any one of claims 1-17, in combination with administering to the patient, concurrently or sequentially, an effective antibacterial amount of a β-lactam antibiotic.
26. compound of Formula I:
Figure imgf000061_0001
(I)
wherein
X is O, CH2, S, SO2, or a direct bond; R1 is CN, SO2CH3, CONH2, CO2H, C02Me, CO(aryl), pyridinyl, thiophenyl, benzothiazolyl, CO-cycloalkyl, or SC -(pyridinyl);
m is 1, 2, 3, or 4;
n is 1, 2, 3, 4, or 5;
each R2 is independently H, halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R2 groups form an ortho-fused methylenedioxy; and
each R3 is independently halo, alkyl, alkoxy, heteroaryl, heterocycle, NRaRb wherein Ra and Rb are each independently H or alkyl, C≡CH, OH, CO2H, C02Me, NO2, OCF3, or SCF3, or two R3 groups form an ortho-fused methylenedioxy;
or a pharmaceutically acceptable salt or solvate thereof;
for preparing a medicament to treat a bacterial infection.
27. The use of claim 26 wherein the medicament further comprises a β-lactam antibiotic.
28. The use of claim 26 or 27 wherein the bacterial infection is a methicillin-resistant Staphylococcus aureus (MRSA) infection.
29. The use of any one of claims 26 or 27 wherein the bacterial infection is a methicillin- resistant Staphylococcus aureus (MRSA) infection and the compound of Formula I is a compound of any one of claims 10-17.
PCT/US2016/067015 2015-12-15 2016-12-15 Potentiators of beta-lactam antibiotics and combination therapy WO2017106552A1 (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4540827A (en) * 1980-11-28 1985-09-10 Bayer Aktiengesellschaft Phenoxybenzene derivatives
US20110077250A1 (en) * 2009-06-26 2011-03-31 Ryder Sean Compounds for modulating rna binding proteins and uses therefor
US20130085162A1 (en) * 2010-03-11 2013-04-04 Dan Littman AMIDO COMPOUNDS AS RORyT MODULATORS AND USES THEREOF

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4540827A (en) * 1980-11-28 1985-09-10 Bayer Aktiengesellschaft Phenoxybenzene derivatives
US20110077250A1 (en) * 2009-06-26 2011-03-31 Ryder Sean Compounds for modulating rna binding proteins and uses therefor
US20130085162A1 (en) * 2010-03-11 2013-04-04 Dan Littman AMIDO COMPOUNDS AS RORyT MODULATORS AND USES THEREOF

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