Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
  • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
  • C07D495/04—Ortho-condensed systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
  • A61K31/4365—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system having sulfur as a ring hetero atom, e.g. ticlopidine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P33/00—Antiparasitic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P33/00—Antiparasitic agents
  • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
  • C07D498/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
  • C07D513/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains three hetero rings
  • C07D513/14—Ortho-condensed systems

Definitions

• the present invention provides 8-methoxy-9H-isothiazolo[5,4-b]quinoline-3,4- diones and related compounds, in which the 7-position substituent is generally an N-linked heterocycloalkyl or heterocycloalkenyl substituent, which possess antimicrobial activity.
• Certain compounds provided herein possess potent antibacterial, antiprotozoal, or antifungal activity.
• Particular compounds provided herein are also potent and/or selective inhibitors of prokaryotic DNA synthesis and prokaryotic reproduction.
• the invention provides anti-microbial compositions, including pharmaceutical compositions, containing one or more carrier, diluents, or excipients.
• the invention provides compounds of Formula I and Formula II (shown below) and includes 8-methoxy-9H-isothiazolo[5,4-b]quinoline-3,4-diones and related compounds of Formula I and II, which possess antimicrobial activity.
• the invention provides compounds of Formula I and Formula EL, which possess potent and/ or selective antibacterial, antiprotozoal, or antifungal activity.
• the invention also provides compositions containing one or more compounds of Formula I or Formula ⁇ , or a salt, solvate, or prodrug, such as an acylated prodrug of such a compound, and one or more carriers, excipients, or diluents.
• the invention further comprises methods of treating and preventing microbial infections, particularly bacterial and protozoal infections by administering an effective amount of a compound of Formula I or Formula H to an animal suffering from or susceptible to a microbial infection.
• microbial infections include bacterial infections, for example E. coli infections, Staphylococcus infections, including Methicillin Resistant Staphylococcus Aureus infections, Salmonella infections and protozoal infections, for example Chlamydia infections.
• the invention is particularly includes methods of preventing or treating microbial infections in mammals, including humans, but also encompasses methods of preventing or treating microbial infections in other animals, including fish, birds, reptiles, and amphibians.
• R 3 is hydrogen, Q-C ⁇ alkyl, C 2 -C 6 alkanoyl, mono- or di-CrC ⁇ alkylcarbamate, or Q-C ⁇ alkylsulfonate; each of which is substituted with O to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, CrQalkoxy, mono- and di-C 1 -C 4 alkylamino, C 1 - C 2 haloalkyl, and C 1 -C 2 haloalkoxy.
• R 6 is hydrogen, halogen, hydroxy, amino, cyano, Q ⁇ alkyl, Q ⁇ alkoxy, mono- or di-(C 1 -C 4 alkyl)amino, -SO 3 R 10 , -SO 2 R 10 , Or -SO 2 NR 10 R 11 .
• R 7 is a nitrogen-linked 6-membered heterocycloalkyl group, 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S, and bridged with a methylene or ethylene bridge.
• (a) is chosen from halogen, hydroxy, amino, nitro, CrC 4 alkyl, Q ⁇ alkoxy, C 1 - C 2 haloalkyl, and C 1 -C 2 haloalkoxy, and
• the compounds of Formula I and Formula II may contain one or more asymmetric elements such as stereogenic centers, stereogenic axes and the like, e.g. asymmetric carbon atoms, so that the compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms. For compounds with two or more asymmetric elements, these compounds can additionally be mixtures of diastereomers.
• the present invention is intended to include all isotopes of atoms occurring in the present compounds.
• Isotopes include those atoms having the same atomic number but different mass numbers.
• isotopes of hydrogen include tritium and deuterium and isotopes of carbon include * C, 13 C, and 14 C.
• an "oxo" substituent on an aromatic group or heteroaromatic group destroys the aromatic character of that group, e.g. a pyridyl substituted with oxo is a pyridone.
• a stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and subsequent formulation into an effective therapeutic agent.
• substituents are named into the core structure. For example, it is to be understood that when (cycloalkyl)alkyl is listed as a possible substituent the point of attachment of this substituent to the core structure is in the alkyl portion.
• alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups, having the specified number of carbon atoms.
• C 1 -C 6 alkyl as used herein includes alkyl groups having from 1 to about 6 carbon atoms.
• Co-C n alkyl is used herein in conjunction with another group, for example, (aryl)C 0 -C 4 alkyl, the indicated group, in this case aryl, is either directly bound by a single covalent bond (C 0 ), or attached by an alkyl chain having the specified number of carbon atoms, in this case from 1 to about 4 carbon atoms.
• alkyl examples include, but are not limited to, methyl, ethyl, n- propyl, isopropyl, n-butyl, t-butyl, n-pentyl, and sec-pentyl.
• alkenyl indicates a hydrocarbon chain of either a straight or branched configuration having one or more carbon-carbon double bond bonds, which may occur at any stable point along the chain.
• alkenyl groups include ethenyl and propenyl.
• Alkynyl indicates a hydrocarbon chain of either a straight or branched configuration having one or more triple carbon-carbon bonds that may occur in any stable point along the chain, such as ethynyl and propynyl.
• Alkoxy represents an alkyl group as defined above with the indicated number of carbon atoms attached through an oxygen bridge.
• alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2-butoxy, t-butoxy, n-pentoxy, 2- pentoxy, 3-pentoxy, isopentoxy, neopentoxy, n-hexoxy, 2-hexoxy, 3-hexoxy, and 3- methylpentoxy.
• An "(alkoxy)alkyl group is an alkoxy group as defined herein attached through its oxygen atom to an alkyl bridge where the point of attachment to the substituted group is in the alkyl group.
• the terms "mono- or di-alkylamino” and “mono- and di- alkylamino” indicate secondary or tertiary alkyl amino groups, wherein the alkyl groups are as defined above and have the indicated number of carbon atoms. The point of attachment of the alkylamino group is on the nitrogen. Examples of mono- and di-alkylamino groups include ethylamino, dimethylamino, and methyl-propyl-amino.
• a mono- or di-(C 3 -C 6 alkyl)(Co- C 4 alkyl)amino group is an alkyl amino substituent in which a first alkyl group is chosen from C 3 - C 6 alkyl and an second alkyl group is chosen from C 0 -C 4 alkyl, wherein C 0 indicates the absence of a second alkyl group, i.e. a mono-Cs-C ⁇ alkylamino.
• alkylester indicates an alkyl group as define above attached through an ester linkage.
• the term "mono-, di-, or tri-alkylhydrazinyl” indicates from 1 to 3 independently chosen alkyl groups as defined above attached through a single-bonded nitrogen-nitrogen linkage. At least one of the alkyl groups is attached to the terminal nitrogen (the nitrogen not bound to the core structure). When the term mono- or di-alkylhydrazinyl is used only the terminal nitrogen is alkyl substituted.
• alkylhydrazinyl groups include 2-butyl-l- hydrazinyl, 2-butyl-2-methyl-l-hydrazinyl, and l,2-dimethyl-2-propyl-l-hydrazinyl.
• alkylthio indicates an alkyl group as defined above attached through a sulfur linkage, i.e. a group of the formula alkyl-S-. Examples include ethylthio and pentylthio.
• aminoalkyl indicates an alkyl group as defined above substituted with at least one amino substituent.
• hydroxyalkyl indicates an alkyl group as defined above, substituted with at least one hydroxyl substituent. In certain instances the alkyl group of the aminoalkyl or hydroxyalkyl group maybe further substituted.
• aryl indicates aromatic groups containing only carbon in the aromatic ring or rings. Typical aryl groups contain 1 to 3 separate, fused, or pendant rings and from 6 to about 18 ring atoms, without heteroatoms as ring members. When indicated, such aryl groups may be further substituted with carbon or non-carbon atoms or groups. Such substitution may include fusion to a 5 to 7-membered saturated cyclic group that optionally contains 1 or 2 heteroatoms independently chosen from N, O, and S, to form, for example, a 3,4- methylenedioxy-phenyl group.
• Aryl groups include, for example, phenyl, naphthyl, including 1- naphthyl and 2-naphthyl, and bi-phenyl.
• (aryl)alkyl aryl and alkyl are as defined above, and the point of attachment is on the alkyl group. This term encompasses, but is not limited to, benzyl, phenylethyl, and piperonyl.
• (aryl)carbohydryl aryl and carbohydryl are as defined above and the point of attachment is on the carbohydryl group, for example a phenylpropen-1-yl group.
• aryl and alkoxy are as defined above and the point of attachment is through the oxygen atom of the alkoxy group; if the alkoxy is a C 0 alkoxy the aryl is attached through an oxygen bridge.
• Carbohydryl as used herein, includes both branched and straight-chain hydrocarbon groups, which are saturated or unsaturated, having the specified number of carbon atoms.
• C 0 -C n carbohydryl is used herein in conjunction with another group, for example, (cycloalkyl)C 0 -C 4 carbohydryl, the indicated group, in this case cycloalkyl, is either directly bound by a single covalent bond (C 0 ), or attached by a carbohydryl chain, such as an alkyl chain, having the specified number of carbon atoms, in this case from 1 to about 4 carbon atoms.
• carbohydryl groups include C 1 -QaIlCyI, such as methyl, or 5-butyl, C 2 -C 6 alkynyl such and hexynyl, and C 2 -C 6 alkenyl, such as 1-propenyl.
• Cycloalkyl indicates a saturated hydrocarbon ring group, having only carbon ring atoms and having the specified number of carbon atoms, usually from 3 to about 8 ring carbon atoms, or from 3 to about 7 carbon atoms.
• Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl as well as bridged or caged saturated ring groups such as norborane or adamantane.
• Cycloalkenyl indicates an unsaturated, but not aromatic, hydrocarbon ring having at least one carbon-carbon double bond. Cycloalkenyl groups contain from 4 to about 8 carbon atoms, usually from 4 to about 7 carbon atoms. Examples include cyclohexenyl and cyclobutenyl.
• (cycloalkyl)alkyl "(cycloalkyl)carbohydryl)," and "(cycloalkyl)alkoxy”
• cycloalkyl, alkyl, carbohydryl, and alkoxy are as defined above, and the point of attachment is on the alkyl, carbohydryl, or alkoxy group respectively.
• These terms include examples such as cyclopropylmethyl, cyclohexylmethyl, cyclohexylpropenyl, and cyclopentylethyoxy.
• (cycloalkenyl)alkyl and “(cycloalkenyl)carbohydryP' the terms cycloalkenyl, alkyl, and carbohydryl are as defined above, and the point of attachment is on the alkyl or carbohydryl group respectively. These terms include examples such as cyclobutenylmethyl, cyclohexenylmethyl, and cyclohexylpropenyl.
• Haloalkyl indicates both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with 1 or more halogen atoms, generally up to the maximum allowable number of halogen atoms.
• haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl.
• Haloalkoxy indicates a haloalkyl group as defined above attached through an oxygen bridge.
• heteroaryl indicates a stable 5- to 7-membered monocyclic or 7- to 10- membered bicyclic heterocyclic ring which contains at least 1 aromatic ring that contains from 1 to 4, or preferably from 1 to 3, heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon. When the total number of S and O atoms in the heteroaryl group exceeds 1, these heteroatoms are not adjacent to one another. It is preferred that the total number of S and O atoms in the heteroaryl group is not more than 2.
• the total number of S and O atoms in the heteroaryl group is not more than 1.
• a nitrogen atom in a heteroaryl group may optionally be quaternized.
• such heteroaryl groups may be further substituted with carbon or non-carbon atoms or groups.
• Such substitution may include fusion to a 5 to 7-membered saturated cyclic group that optionally contains 1 or 2 heteroatoms independently chosen from N, O, and S, to form, for example, a [l,3]dioxolo[4,5-c]pyridyl group.
• heteroaryl groups include, but are not limited to, pyridyl, indolyl, pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl, and 5,6,7,8-tetrahydroisoquinoline.
• heteroaryl, alkyl, and carbohydryl are as defined above, and the point of attachment is on the alkyl or carbohydryl group respectively.
• heteroaryl, alkyl, and carbohydryl include such examples as pyridylmethyl, thiophenylmethyl, and (pyrrolyl) 1 -ethyl.
• heterocycloalkyl indicates a saturated cyclic group containing from 1 to about 3 heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon. Heterocycloalkyl groups have from 3 to about 8 ring atoms, and more typically have from 5 to 7 ring atoms. Examples of heterocycloalkyl groups include morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl groups. A nitrogen in a heterocycloalkyl group may optionally be quaternized.
• heterocyclic group indicates a 5-6 membered saturated, partially unsaturated, or aromatic ring containing from 1 to about 4 heteroatoms chosen from N, O, and S, with remaining ring atoms being carbon or a 7-10 membered bicyclic saturated, partially unsaturated, or aromatic heterocylic ring system containing at least 1 heteroatom in the two ring system chosen from N, O, and S and containing up to about 4 heteroatoms independently chosen from N, O, and S in each ring of the two ring system.
• the heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure.
• heterocyclic rings described herein maybe substituted on carbon or on a nitrogen atom if the resulting compound is stable.
• a nitrogen atom in the heterocycle may optionally be quaternized. It is preferred that the total number of heteroatoms in a heterocyclic groups is not more than 4 and that the total number of S and O atoms in a heterocyclic group is not more than 2, more preferably not more than 1.
• heterocyclic groups include, pyridyl, indolyl, pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl, isoindolyl, dihydroisoindolyl, 5,6,7,8-tetrahydroisoquinoline, pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, and pyrrolidin
• heterocyclic groups include, but are not limited to, phthalazinyl, oxazolyl, indolizinyl, indazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzoisoxolyl, dihydro-benzodioxinyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl, imidazopyridinyl, isothiazolyl, naphthyridinyl, cinnolinyl, carbazolyl, beta- carbolinyl, isochromanyl, chromanonyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazoly
• Active agents are compounds that have pharmaceutical utility, e.g. may be used to treat a patient suffering from a disease or condition, or may be used prophylactically to prevent the onset of a disease or condition in a patient , or that may be used to enhance the pharmaceutical activity of other compounds.
• compositions are compositions comprising at least one active agent, such as a compound or salt of Formula I or Formula II, and at least one other substance, such as a carrier, excipient, or diluent.
• Pharmaceutical compositions meet the U.S. FDA's GMP (good manufacturing practice) standards for human or non-human drugs.
• Salts of the compounds of the present invention include inorganic and organic acid and base addition salts.
• the salts of the present compounds can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two.
• the appropriate base such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or the like
• salts of the present compounds further include solvates of the compounds and of the compound salts.
• “Pharmaceutically acceptable salts” include derivatives of the disclosed compounds wherein the parent compound is modified by making non-toxic acid or base salts thereof, and further refers to pharmaceutically acceptable solvates of such compounds and such salts.
• pharmaceutically acceptable salts include, but are not limited to, 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 the conventional non-toxic salts and the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
• conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2- acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, HOOC-(CH 2 ) n -COOH where n is 0-4, and the like. Lists of additional suitable salts may be found, e.g., in Remington's Pharmaceutical Sciences, 17
• prodrugs includes any compounds that become compounds of Formula I when administered to a mammalian subject, e.g., upon metabolic processing of the prodrug.
• prodrugs include, but are not limited to, acetate, formate and benzoate and like derivatives of functional groups (such as alcohol or amine groups) in the compounds of Formula I and Formula II.
• a therapeutically effective amount of a compound of this invention means an amount effective, when administered to a human or non-human patient, to provide a therapeutic benefit such as an amelioration of symptoms, e.g., an amount effective to decrease the symptoms of a microbial infection, and or an amount sufficient to reduce the symptoms of a bacterial, fungal, or protozoal infection. In certain circumstances a patient suffering from a microbial infection may not present symptoms of being infected. Thus a therapeutically effective amount of a compound is also an amount sufficient to prevent a significant increase or significantly reduce the detectable level of microorganism or antibodies against the microorganism in the patient's blood, serum, other bodily fluids, or tissues.
• the invention also includes using compounds of Formula I and Formula II in prophylactic therapies.
• a "therapeutically effective amount” is an amount sufficient to significantly decrease the treated animal's risk of contracting a microorganism infection.
• a significant reduction is any detectable negative change that is statistically significant, for example statistical significance can be measured in a standard parametric test of statistical significance such as Student's T-test, where p ⁇ 0.05.
• the numbers 1 through 9 refer specifically to positions within the tricyclic ring system whereas the letters A, B and C refer to the specific six (rings A and B) or five (ring C) member rings as shown below.
• the invention also includes compounds of Formula I and Formula II and the pharmaceutically acceptable salts thereof, in which the 8-position of the tricyclic structure is not required to be an aromatic carbon atom substituted with a methoxy, but is A 8 .
• R 3 is hydrogen, Ci-C 6 alkyl, C 2 -C 6 alkanoyl, mono- or di-Ci ⁇ C 6 alkylcarbamate, or Ci-C 6 alkylsulfonate; each of which is substituted with O to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, Q-C 4 alkoxy, mono- and di-Ci-C 4 alkylamino, Q- C 2 haloalkyl, and Ci-C 2 haloalkoxy.
• R 7 is a nitrogen-linked heterocycloalkyl or heterocycloalkenyl group, each of which has 4 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S, each of which heterocycloalkyl or heterocycloalkenyl group is substituted with at least one group (ii) and is substituted with 0 or 1 or more of (i) and (iii).
• R 7 is a nitrogen-linked Ci-C 4 alkylamino substituted with a 5 or 6-membered heteroaryl group having 1 or 2 heteroatoms independently chosen from N, O, and S, or substituted with a heterocycloalkyl group or heterocycloalkenyl group, each of which has 4 to 8 ring members, including 1 or 2 ring heteroatoms independently chosen from N, O, and S, each of which heteroaryl, heterocycloalkyl, or heterocycloalkenyl group is substituted with 0 or 1 or more substituents independently chosen from (i), (ii), and (iii).
• R 7 is a nitrogen-linked heterocycloalkyl group, having 5 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S, and bridged with a methylene or ethylene bridge, with the proviso that when R 7 is a 2,5 methylene bridged piperazine it is substituted with at least one group (ii) or (iii).
• a 8 is N or CR 8 .
• R 8 is hydrogen, halogen, hydroxy, amino, cyano, nitro, or -NHNH 2 .
• R 9 is CrC 8 alkyl, (C 3 -C 7 cycloalkyl)C 0 -C 4 alkyl, or phenyl, each of which is substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, -COOH, -CONH 2 , C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C r C 4 alkoxy, (C 3 - C 7 cycloalkyl)C 0 -C 4 alkyl, (C 3 -C 7 cycloalkyl)Co-C 4 alkoxy, mono- and di-(C 1 -C 4 alkyl)amino, C 1 - C 2 haloalkyl, Q-C ⁇ aloalkoxy, and C 2 -C 4 alkanoyl.
• the invention further provides compounds and salts of Formula I and Formula II in which the variables (e.g. A 1 , R 2 , R 3 , R 4 , etc.) carry definitions other than those set forth above.
• the variables e.g. A 1 , R 2 , R 3 , R 4 , etc.
• Embodiments in which one or more of the following conditions is met are included in the invention:
• a 1 is S; e.g. compounds and salts of Formula III and Formula IV are included herein.
• a 1 is SO 2 ; e.g. compounds and salts of Formula VII and VIII are included herein.
• the R. 2 variable (compounds and salts of Formula I):
• R 3 is Ci-C 2 alkyl.
• R 5 is hydrogen, amino, mono- or di-(C 1 -C 2 )alkylamino, or mono- or di-Ci-C 2 alkylhydrazinyl.
• R 6 is fluoro or hydrogen.
• R 6 is fluoro
• R 7 is a nitrogen-linked Ci-C 4 alkylamino substituted with a 5 or 6-membered heteroaryl group having 1 or 2 heteroatoms independently chosen from N, O, and S, or substituted with a heterocycloalkyl group, heterocycloalkenyl group, each of which has 4 to 8 ring members, including 1 or 2 ring heteroatoms independently chosen from N, O, and S, each of which heteroaryl, heterocycloalkyl, or heterocycloalkenyl group is substituted with 0 or 1 or more substituents independently chosen from (i), (ii), and (iii); or
• R 7 is a nitrogen-linked heterocycloalkyl or heterocycloalkenyl group, each of which has 4 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N,
• R 7 is a nitrogen-linked heterocycloalkyl group, having 5 to 8 ring members, including 0,
• (i) is chosen from halogen, hydroxy, amino, nitro, Ci-C 2 alkyl, mono- and di-(Cr C 2 )alkylamino, and -CH 2 CH 2 F;
• (ii) is chosen from cyano, oxo, C 3 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, (C 1 -C 4 alkoxy)C 0 - C 4 alkyl, mono- and di-(C 3 -C 6 alkyl)(C 0 -C 4 alkyl)amino, di-(C 1 -C 4 alkyl)aminoC 1 -C 4 alkyl, mono-or di-alkylamino(branched C 2 -C 4 alkyl), (C 3 -C 7 cycloalkylamino)C 1 -C 4 alkyl, Q-Qhaloalkyl other than -CH 2 CH 2 F, C r C 2 haloalkoxy, (C 3 -C 7 cycloalkyl)C 0 -C 2 alkyl, (C 3 -C 7 cycloalkyl)C 0 -C 2
• each of (ii) and (iii) is substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, oxo, -COOH 5 -CONH 2 , C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Q ⁇ alkoxy, (C 3 -C 7 cycloalkyl)C 0 - C 2 alkyl, (C 3 -C 7 cycloalkyl)CQ-C 2 alkylamino, (C 3 -C 7 cycloalkyl)C 0 -C 4 alkoxy, mono- and di-(Cr C 4 alkyl)amino, Q-Qhaloalkyl, C 1 -C 2 haloalkoxy, C 2 -C 4 alkanoyl, and phenyl.
• substituents independently chosen from halogen, hydroxy, amino, cyano, nitro
• R 7 is a nitrogen-linked heterocycloalkyl or heterocycloalkenyl group, each of which has 4- to 8- ring members, including 0, 1, or 2 additional ring heteroatoms independently chosen from N, O, and S, each of which is substituted with at least one group (ii) and is substituted with 0 or 1 or more of (i) and (iii).
• (i) is chosen from halogen, hydroxy, amino, nitro, Q-Qalkyl, mono- and di-(Q- C 2 )alkylamino, and -CH 2 CH 2 F;
• (ii) is chosen from oxo and cyano, C 3 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, (C 1 - C 4 alkoxy)Co-C 4 alkyl, mono- and di-(C 3 -C 6 alkyl)(C 0 -C 4 alkyl)amino, di-(C 1 -C 4 alkyl)aminoC 1 - C 4 alkyl, mono-and di-alkylamino(branched C 2 -C 4 alkyl), (C 3 -C 7 cycloalkylamino)C 1 -C 4 alkyl, C 1 - C 2 haloalkyl other than -CH 2 CH 2 F, Q-C ⁇ aloalkoxy, (C 3 -C 7 cycloalkyl)C 0 -C 2 alkyl, (C 3 - C 7 cycloalkyl)C 0 -C 2 al
• each of (ii) other than oxo and cyano and each of (iii) is substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, oxo, -COOH, -CONH 2 , Q-Qalkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 alkoxy, (C 3 - C 7 cycloalkyl)C 0 -C 2 alkyl, (C 3 -C 7 cycloalkyl)C 0 -C 2 alkylamino, (C 3 -C 7 cycloalkyl)C 0 -C 4 alkoxy, mono- and di-(C 1 -C 4 alkyl)amino, Q-Qjhaloalkyl, Q-C ⁇ aloalkoxy, C 2 -C 4 alkanoyl, and phenyl.
• substituents independently chosen from
• R 7 is a 4-, 5-, or 6-membered nitrogen-linked heterocycloalkyl, which has 0 or 1 additional nitrogen atoms, which 4-, 5-, or 6-membered nitrogen-linked heterocycloalkyl is substituted with 0 to 2 substituents independently chosen from (i) and one substituent (ii).
• (i) is chosen from halogen, hydroxy, amino, C 1 -C 2 alkyl, hydroxyC 1 -C 2 aUcyl, aminoCr C 2 alkyl, and cyano, and
• each of (ii) other than oxo and cyano is substituted with substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, oxo, -C 1 -QaIlCyI, C 2 -C 4 alkenyl, Ci-C 4 alkoxy, (C 3 -C 7 cycloalkyl)C 0 -C 2 alkyl, (C 3 - C 7 cycloalkyl)C 0 -C 2 alkylamino, mono- and di-(C 1 -C 4 alkyl)amino, CrQhaloalkyl, and C 1 - C 2 haloalkoxy.
• substituents independently chosen from halogen, hydroxy, amino, cyano, oxo, -C 1 -QaIlCyI, C 2 -C 4 alkenyl, Ci-C 4 alkoxy, (C 3 -C 7 cycloalkyl)C 0
• R 7 is a pyrrolidinyl, piperidinyl, piperazinyl, mo ⁇ holinyl, thiomorpholinyl, or azepanyl group substituted with at least one group (ii) and substituted with 0 or one or more of (i) and (iii).
• (i) and (iii) carry any of the definitions set forth above.
• R 7 is a pyrrolidinyl or piperazinyl group substituted with one group (ii) and 0, 1, or 2 substituents independently chosen from halogen, hydroxy, trifluoromethyl, or trifluoromethoxy, wherein (ii) is oxo, cyano, C 2 -C 4 alkanoyl or (ii) C 3 -C 7 cycloalkyl substituted with 0 or 1 C 1 -C 2 alkyl or amino substituents, or (ii) is C 3 -C 6 alkyl, di-(C 1 -C 4 alkyl)aminoC 1 - C 4 alkyl, mono-or di-alkylamino(branched C 2 -C 4 alkyl), or (C 3 -C 7 cycloalkylamino)C 1 -C 4 alkyl, each of which is substituted with 0, 1 or 2 substituents independently chosen from amino, hydroxy, (C 3 -C 7 cycl
• R 7 is a pyrrolidinyl or piperazinyl group substituted with one group (ii) and optionally substituted with 1 methyl or halogen substituent wherein (ii) is C 3 -C 7 cycloalkyl which is unsubstituted or substituted with one amino substituent, or (ii) is C 3 -C 6 alkyl substituted with 1 amino, hydroxy, C 3 -C 7 cycloalkyl, or (C 3 -C 7 cycloalkyl)Co-C 2 alkylamino substituent, or (ii) is or di-(C 1 -C 4 alkyl)aminoC 1 -C 4 alkyl, mono-or di-alkylamino(branched C 2 -C 4 alkyl) or (C 3 - C 7 cycloalkylamino)C 1 -C 4 alkyl, each of which is substituted with 0 or 1 C 3 -C 7 cycloalkyl
• R 7 is a group of formula (a) - (e)
• R 15 is (ii); and R 16 is 0 or 1 or more substituents independently chosen from chloro, fluoro, methyl, methoxy, aminomethyl, aminoethyl, trifluoromethyl, and trifluoromethoxy.
• R 7 is a group of formula (a) - (e) in which
• R 15 is oxo or cyano
• R 15 is C 3 -C 7 cycloalkyl substituted with 0 or 1 C 1 -C 2 alkyl or amino substituent, or
• R 15 is C 3 -C 6 alkyl, di-(C 1 -C 4 alkyl)aminoC 1 -C 4 alkyl, mono-or di-alkylamino(branched C 2 - C 4 alkyl), or (C 3 -C 7 cycloalkylamino)C 1 -C 4 alkyl, each of which is substituted with 0, 1, or 2 substituents independently chosen from amino, hydroxy, and (C 3 -C 7 cycloalkyl)Co-C 2 alkyl; or
• R 15 is mono- or di-(C 3 -C 6 alkyl)(Co-C 4 alkyl)amino, or
• R 7 is a group of formula (a) - (e) in which
• R 15 is C 3 -C 7 cycloalkyl which is unsubstituted or substituted with one amino substituent, or R 15 is C 3 -C O alkyl, di-(C 1 -C 4 alkyl)aminoC 1 -C 4 alkyl, mono-or di-alkylamino(branched C 2 - Qalkyl) or (C 3 -C 7 cycloalkylamino)C 1 -C 4 alkyl, each of which is substituted with 0, 1, or 2 substituents independently chosen from amino, hydroxy, and (C 3 -C 7 cycloalkyl)C 0 -C 2 alkyl.
• R 16 is 0 or 1 substituent chosen from chloro, fluoro, and methyl.
• R 7 is a group of formula (a) - (e) in which
• R 15 is cyclopropyl substituted with amino, or R 15 is C 3 -C 6 alkyl substituted with amino or C 3 -C 5 cycloalkylamino, or R 15 is mono- or di-(C 1 -C 4 alkyl)aminoC 1 -C 4 alkyl.
• R 16 is 0 or 1 substituent chosen from chloro, fluoro, and methyl.
• R 7 is a group of the formula
• R 7 is a nitrogen-linked C 1 -C 4 alkylamino substituted with a 5 or 6-membered heteroaryl group having 1 or 2 heteroatoms independently chosen from N, O, and S, or substituted with a heterocycloalkyl group, heterocycloalkenyl group, each of which has 4 to 8 ring members, and 1 or 2 ring heteroatoms independently chosen from N, O, and S, each of which heteroaryl, heterocycloalkyl, or heterocycloalkenyl group is substituted with 0 or 1 or more substituents independently chosen from (i), (ii), and (iii).
• substituents independently chosen from (i), (ii), and (iii).
• (i), (ii), and (iii) carry any of the definitions set forth above.
• R 7 is an N-linked Q-Qalkylamino substituted with a 5- or 6-membered heteroaryl group having 1 or 2 heteroatoms independently chosen from N, O, and S, or substituted with a 5- or 6- membered heterocycloalkyl having 1 or 2 ring heteroatoms independently chosen from N, O, and S, each of which heteroaryl or heterocycloalkyl is substituted with 0, 1, or 2 substituents independently chosen from halogen, hydroxy, amino, oxo, cyano, Ci-C 2 alkyl, C 1 -C 2 alkoxy, Q-C ⁇ aloalkyl, and Q-Qhaloalkoxy.
• R 7 is Q-Qalkylamino substituted with a pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, thienyl, pyrrolidinyl, furanyl, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, or pyrrolidinyl, each of which is substituted with 0, 1, or 2 substituents independently chosen from halogen, hydroxy, amino, oxo, cyano, Ci-C 2 alkyl, C 1 -C 2 alkoxy, Q-C ⁇ aloalkyl, and C 1 - C 2 haloalkoxy.
• R 7 is C 1 -C 2 alkylamino substituted with pyridyl, piperazinyl, piperidinyl, or morpholinyl, each of which is substituted with 0, 1, or 2 substituents independently chosen from halogen, methyl, and methoxy.
• R 7 is a nitrogen-linked heterocycloalkyl or heterocycloalkenyl group, each of which has 4 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S, forming part of a bicyclic system with a 3- to 7-membered carbocyclic or heterocyclic ring in fused or spiro orientation, and is substituted with 0 or 1 or more substituents independently chosen from (i), (ii), and (iii).
• substituents independently chosen from (i), (ii), and (iii).
• (i), (ii), and (iii) carry any of the definitions set forth above.
• R 7 is a group of the formula
• R 7 is a 5- or 6-membered nitrogen-linked heterocycloalkyl, which has 0 or 1 additional nitrogen atoms, which 5- or 6-membered nitrogen-linked heterocycloalkyl is part of a bicyclic ring system having a fused Cs-C ⁇ cycloalkyl or a fused 4- to 6- membered heterocycloalkyl containing 1 nitrogen atom, which bicyclic ring system is substituted with 0 to 3 substituents independently chosen from (i) and (ii).
• (i) is chosen from halogen, hydroxy, and amino, and
• Each of (ii) is substituted with substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, oxo, - C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 1 -C 4 alkoxy, (C 3 -C 7 cycloalkyl)Co-C 2 alkyl, mono- and di-(C 1 -C 4 alkyl)amino, Q-Qhaloalkyl, and Q-Qhaloalkoxy.
• substituents independently chosen from halogen, hydroxy, amino, cyano, oxo, - C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 1 -C 4 alkoxy, (C 3 -C 7 cycloalkyl)Co-C 2 alkyl, mono- and di-(C 1 -C 4 alkyl)amino, Q-Qhaloalkyl, and Q-Qhaloalkoxy
• the 5- or 6-membered nitrogen-linked heterocycloalkyl which is part of a bicyclic ring system is a pyrrolidinyl or piperidinyl and is fused to a C 3 - C 6 cycloalkyl, pyrrolidinyl, or piperidinyl; in which the bicyclic ring is substituted with 0, 1, or 2 substituents independently chosen from halogen, methyl, or methoxy.
• the bicyclic ring system is a 3-aza-bicyclo[3.1.0]hexanyl or a octahydro-lH-pyrrolo[3,4-b]pyridinyl ring system.
• R 7 is a nitrogen-linked heterocycloalkyl group, having 5 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S, and bridged with a methylene or ethylene bridge, with the proviso that when R 7 is a 2,5 methylene bridged piperazine it is substituted with at least one group (ii) or (iii) (other R 7 groups of this embodiment are optionally substituted with on ore more of (i), (ii), and (iii)). Wherein (i), (ii), and (iii) carry any of the definitions set forth above.
• R 7 is a bridged piperidinyl or bridged piperazinyl, each of which is substituted with 0 to 3 substituents independently chosen from (i) and (ii).
• Each of (ii) other than oxo and cyano is substituted with 0 to 3 substituents independently chosen from halogen, hydroxy, amino, cyano, oxo, C 1 -C 4 aUcyl, C 2 -C 4 alkenyl, Q-Qalkoxy, (C 3 -C 7 cycloalkyl)Co-C 2 cycloalkyl, mono- and di- ⁇ Q-QalkyOamino, Q-Qhaloalkyl, and C 1 -C 2 ImIOaIkOXy.
• R 7 is a bridged piperidinyl or bridged piperazinyl, each of which is substituted with 0 to 2 substituents independently chosen from halogen, hydroxy, amino, cyano, C 1 -C 2 alkyl, and Q-Caalkoxy.
• R 8 is methoxy
• R 7 is a nitrogen-linked heterocycloalkyl group, which has 4 to 8 ring members, including 0, 1 or 2 additional ring heteroatoms independently chosen from N, O, and S which heterocycloalkyl, which R 7 is substituted with 0 or 1 or more substituents independently chosen from (a) and 0 or 1 substituents chosen from (b), wherein
• (a) is chosen from halogen, hydroxy, amino, nitro, C 1 -QaI]CyI, C 1 -C 4 alkoxy, C 1 - C 2 haloalkyl, Q-C ⁇ aloalkoxy,
• (b) is oxo, amino, cyano, hydroxyC 1 -C 4 alkyl, aminoC 1 -C 4 alkyl, Ci-C ⁇ alkylthio, C 2 - Cgalkanoyl, (mono- or di-C 1 -C 4 alkyl)aminoC 0 -C 4 alkyl, (C 3 -C 7 cycloalkyl)C 0 -C 4 alkyl, (C 3 - C 7 cycloalkylamino)C 0 -C 4 alkyl, (C 3 -C 7 cycloaUcyl)(C 1 -C 4 alkyl)aminoCo-C 4 alkyl, (heterocycloalkyl)Co-C 4 alkyl, or (aryl)C 0 -C 4 alkyl.
• Each of (b) other than oxo and cyano is substituted with 0 to 2 substituents independently chosen from halogen, hydroxy, amino, cyano, nitro, oxo, -COOH, -CONH 2 , C 1 -C 4 alkyl, C 2 - C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 alkoxy, mono- and di-(C 1 -C 4 alkyl)amino, d-Qhaloalkyl, and Q-Cyialoalkoxy.
• R 8 is methoxy
• R 7 is a A-, 5-, or 6-membered nitrogen-linked heterocycloalkyl, which has 0 or 1 additional N, S, or O atoms, which 4-, 5-, or 6-membered nitrogen-linked heterocycloalkyl is substituted with 0 to 2 substituents independently chosen from (a) and 0 or 1 substituent (b), wherein (a) and (b) carry the definitions set forth above.
• R 8 is methoxy
• R 7 is a pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, or azepanyl group substituted with substituted with 0 to 2 substituents independently chosen from or more of (a) and 0 or 1 substituents (b), wherein (a) and (b) carry the definitions set forth above.
• R 8 is methoxy
• R 7 is a piperazinyl or thiomorpholinyl group, each of which is substituted with 0 to 2 substituents independently chosen from or more of (a) and 0 or 1 substituents (b); wherein
• (a) is chosen from halogen, hydroxy, amino, Q-Otalkyl, C 1 -C 2 alkoxy, Ci-Qhaloalkyl, and CrQhaloalkoxy, and
• (b) is oxo, amino, cyano, hydroxyC 1 -C 4 alkyl, aminoC 1 -C 4 alkyl, C 2 -C 4 alkanoyl (mono- and di-C 1 -C 4 alkyl)aminoC 0 -C 4 alkyl, (C 3 -C 7 cycloalkyl)Co-C 4 alkyl substituted with amino, (C 3 - C 7 cycloalkylamino)Co-C 4 alkyl, or (C 3 -C 7 cycloalkyl)(C 1 -C 4 alkyl)aminoCo-C 4 alkyl.
• R 8 is methoxy
• R 7 is a pyrrolidinyl group, which is substituted with 0 to 2 substituents independently chosen from or more of (a) and 0 or 1 substituents (b), wherein (a) and (b) may carry any of the definitions set forth above for these variables.
• R 8 is methoxy
• R 7 is pyrrolidinyl group substituted with one group (b) and optionally substituted with 1 methyl or halogen substituent wherein (b) is oxo, amino, cyano, hydroxyC 1 -C 4 alkyl, aminoCi- C 4 alkyl, C 2 -C 4 alkanoyl (mono- and di-Q ⁇ alky ⁇ aminoCo-Qalkyl, (C 3 -C 7 cycloalkyl)C 0 - C 2 alkyl substituted with amino, (C 3 -C 7 cycloalkylamino)C 0 -C 4 alkyl, or (C 3 -C 7 CyClOaIlCyI)(C 1 - C4alkyl)aminoCo-C 4 alkyl.
• R 15 is (b); and R 16 is 0 or 1 or more substituents chosen from amino, hydroxy, chloro, fluoro, methyl, methoxy, trifluoromethyl, and trifluoromethoxy; (b) may carry any of the definitions set forth above for these variables.
• R 15 is oxo, amino, cyano, hydroxyC 1 -C 4 alkyl, aminoC 1 -C 4 alkyl, C 2 -C 4 alkanoyl, (mono- and di-Cr C 4 alkylamino)C 0 -C 4 alkyl 5 (C 3 -C 7 cycloalkyl)C 0 -C 2 alkyl substituted with amino, (C 3 - C 7 cycloalkylamino)Co-C 4 alkyl, or (C 3 -C 7 cycloalkyl)(C 1 -C 4 alkyl)aminoC 0 -C 4 alkyl.
• R 15 is oxo, cyano, hydroxyQ-G t alkyl, aminoC 1 -C 4 alkyl, acetyl, (mono- and di- C 1 -C 2 alkylamino)C 1 -C 4 alkyl, cyclopropyl substituted with amino, or (C 3 -C 7 cycloalkylamino)Co- C 4 alkyl; and R 16 is 0 or 1 substituent chosen from hydroxy, amino, chloro, and methyl.
• R 7 is a group of the formula
• R 8 is methoxy
• R 7 is a nitrogen-linked C t -Qalkylamino substituted with a 5 or 6-membered heteroaryl group having 1 or 2 heteroatoms independently chosen from N, O, and S, or substituted with a heterocycloalkyl group, which has 4 to 8 ring members, including 1 or 2 ring heteroatoms independently chosen from N, O, and S;
• R 7 is substituted with 0 or 1 or more substituents independently chosen from (a) and 0 or 1 substituents chosen from (b), wherein (a) and (b) may carry any of the definitions set forth above for these variables.
• R 8 is methoxy
• R 7 is substituted with 0 or 1 or more substituents independently chosen from (a) and 0 or 1 substituents chosen from (b), wherein (a) and (b) carry any of the definitions set forth above for these variables.
• R 7 is octahydropyrrolo[3,4-b]pyridin-6-yl ring system which is substituted with 0, 1, or 2 substituents independently chosen from halogen, methyl, or methoxy.
• a 8 is CR 8 , and R 8 is hydrogen, halogen, C 1 -C 2 alkyl, C 1 -C 2 alkoxy, trifluoromethyl, or trifluoromethoxy.
• R 9 is ethyl, t-butyl, cyclopropyl, or 2,4-difluorophenyl.
• R 5 is hydrogen (3).
• R 15 is mono- or di-(C 3 -C 6 alkyl)(C 0 -C 4 alkyl)amino, or
• Certain compounds of Formula I and Formula II are selective antimicrobial agents; having the ability to kill or inhibit the growth or reproduction of microbial organisms, while having little or no effect on the cells of fish, amphibians, reptiles, birds, or mammals.
• the selectivity of compounds of Formula I and Formula II may be assessed by determining the CC 50 (the concentration at which 50% of the cells are killed) for cultured cells of a higher animal, such as a fish, reptiles, amphibian, bird, or mammal.
• Certain compounds of the invention exhibit a CC 50 of greater that 100 micromolar for mammalian cells.
• the MRSA strain used in this study was ATCC 700699, from ATTC, Manassas VA.
• Hep2 cells are ATCC catalog number CCL-23, Manassas VA.
• the invention provides pharmaceutical compositions comprising a compound or salt of salt thereof of Formula I or Formula II, together with a pharmaceutically acceptable carrier, diluent, or excipient.
• the pharmaceutical composition may be formulated as any pharmaceutically useful form, e.g., as an aerosol, a cream, a gel, a pill, a capsule, a tablet, a syrup, a transdermal patch, or an ophthalmic solution.
• Optional active agents maybe included in a pharmaceutical composition, which do not substantially interfere with the activity of the compound of the present invention.
• compositions containing compounds of general Formula I and/ or Formula II may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs.
• Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents, such as sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide pharmaceutically elegant and palatable preparations.
• Oral formulations contain between 0.1 and 99% of a compound of the invention and usually at least about 5% (weight %) of a compound of the present invention. Some embodiments contain from about 25% to about 50% or from 5% to 75 % of a compound of invention. Liquids formulations
• styrene foams include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water.
• Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent.
• Suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water.
• Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent.
• Compounds of the invention may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intracisternal or intraspinal application.
• Topical compositions of the present invention may be in any form including, for example, solutions, creams, ointments, gels, lotions, milks, cleansers, moisturizers, sprays, skin patches, and the like.
• Such solutions may be formulated as 0.01% -10% isotonic solutions, pH about 5-7, with appropriate salts.
• Compounds of the invention may also be formulated for transdermal administration as a transdermal patch.
• compositions suitable for use in topical carriers include, for example, emollients, solvents, humectants, thickeners and powders. Examples of each of these types of materials, which can be used singly or as mixtures of one or more materials, are as follows:
• Emollients such as stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane- 1,2-diol, butane- 1,3-diol, mink oil, cetyl alcohol, iso-propyl isostearate, stearic acid, iso-butyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, dimethylpolysiloxane, di-n-butyl sebacate, iso-propyl myristate, iso-propyl palmitate, iso-propyl stearate, butyl stearate, polyethylene glycol, Methylene glycol, lanolin, sesame oil, coconut oil, arachis oil, castor oil, ace
• compositions of the present invention may also optionally comprise an activity enhancer.
• the activity enhancer can be chosen from a wide variety of molecules that function in different ways to enhance antimicrobial effects of compounds of the present invention.
• Particular classes of activity enhancers include skin penetration enhancers and absorbtion enhancers.
• the invention includes packaged pharmaceutical formulations.
• packaged formulations include a pharmaceutical composition containing one or more compounds or salts of Formula I or Formula II in a container and instructions for using the composition to treat an animal (typically a human patient) suffering from a microorganism infection) or prevent a microorganism infection in an animal.
• compositions can be administered alone or as mixtures, and the compositions may further include additional drugs or excipients as appropriate for the indication.
• the invention includes methods of preventing and treating microorganism infections, particularly bacterial and protozoal infections, by administering an effective amount of one or more compounds of Formula I and of Formula II to an animal at risk for a microorganism infection or suffering from a microorganism infection.
• the animal may be a fish, amphibian, reptile or bird, but is preferably a mammal.
• microorganism infections in livestock animals, companion animals, and human patients are particularly preferred.
• the compounds disclosed herein are useful for preventing and treating bacterial infections in animals. Furthermore compounds of the invention may be used to treat a variety of conditions not attributed to bacterial infections. These include diseases and disorders caused fungal infections, mycoplasma infections, protozoal infections, or other conditions involving infectious organisms.
• Methods of treatment also include inhibiting microorganism replication in vivo, in an animal at risk for a microorganism infection or suffering from such an infection, by administering a sufficient concentration of a compound of Formula I or Formula II to inhibit bacterial survival in vitro.
• a sufficient concentration of a compound administered to the patient is meant the concentration of the compound available in the animal's system to prevent or combat the infection.
• concentration by be ascertained experimentally, for example by assaying blood concentration of the compound, or theoretically, by calculating bioavailability.
• the amount of a compound sufficient to inhibit bacterial survival in vitro may be determined with a conventional assay for bacterial survival such as the Minimum Inhibitory Concentration (MIC) Assay disclosed in Example 9, which follows.
• MIC Minimum Inhibitory Concentration
• the invention also includes using compounds of Formula I and Formula I in prophylactic therapies.
• an effective amount of a compound of the invention is an amount sufficient to significantly decrease the treated animal's risk of contracting a microorganism infection.
• Compounds of the invention are particularly useful for treating and preventing infectious disorders. These include for example: ocular infections such as conjunctivitis; urinary tract and genital infections, such as complicated urinary tract infections, acute urinary tract and 29302
• genital infections such as pyelonephritis, cervical gonococcal infections, cystitis, urethral chlamydial infections, cervical chlamydial infections, urethral gonococcal infections, and prostatitis
• respiratory infections such as lower respiratory tract infections, acute sinusitis, acute exacerbations of chronic bronchitis, community-acquired pneumonia, and nosocomial pneumonia
• skin infections such as skin-structure infections, impetigo, folliculitis, boils, scalded skin syndrome, and cellulites
• other infections such as bone infections, joint infections, infectious diarrhea, typhoid fever, intra-abdominal infections, gynecologic infections, including toxic shock syndrome, pelvic infections, and post-surgical infections.
• the disclosed compounds are useful for treating infections caused by the following microorganisms:
• Aerobic Gram-positive Microorganisms Including but not limited to Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus (including methicillan S, aureus), Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus haemolyticus, and Staphylococcus hominis.
• Aerobic Gram-negative Microorganisms Including but not limited to Campylobacter jejuni, Citrobacter diversus, Citrobacter freundii, Enterobacter cloacae, Escherichia coli, Haemophilus influenzae, Haemophilus parainfluenzae, Klebsiella pneumoniae, Moraxella catarrhalis, Morganella morganii, Neisseria gonorrhoeae, Proteus mirabilis, Proteus vulgaris, Providencia rettgeri, Providencia stuartii, Pseudomonas aeruginosa, Stenotrophomonas maltophila, Salmonella typhi, Serratia marcescens, Shigella boydii, Shigella dysenteriae, Shigella flexneri, Shigella sonnei.
• Dosage levels of the order of from about 0.1 mg to about 140 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions (about 0.5 mg to about 7 g per patient per day).
• the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Dosage unit forms will generally contain between from about 1 mg to about 500 mg of an active ingredient.
• Frequency of dosage may also vary depending on the compound used and the particular disease treated. However, for treatment of most infectious disorders, a dosage regimen of 4 times daily or less is preferred and a dosage regimen of.l or 2 times daily is particularly preferred.
• the compounds of the invention may also be useful in combination with other pharmaceutically active agents such as antibacterial agents, antiviral agents, antifungal agents, antiinflammatories, interferon, efflux-pump inhibitors, and beta-lactamase inhibitors.
• Antibiotic agents include any molecule that tends to prevent, inhibit or destroy life and as such, includes anti-bacterial agents, anti-fungicides, anti-viral agents, and anti-parasitic agents.
• compositions of the invention include single dosage forms containing of a compound of Formula I and/or Formula II and one or more other active agent, dosage forms containing more than one compound of Formula I and/ or Formula ⁇ , and separate administration of a compound of Formula I and/or Formula II with another active agent.
• active agents which are useful in combinations of the invention, may be isolated from an organism that produces the agent or synthesized by methods known to those of ordinary skill in the art of medicinal chemistry or purchased from a commercial source.
• Anti-bacterial antibiotic agents include, but are not limited to, penicillins, cephalosporins, carbacephems, cephamycins, carbapenems, monobactams, aminoglycosides, glycopeptides, quinolones, tetracyclines, macrolides, and fluoroquinolones (see Table below).
• antibiotic agents include, but are not limited to, Penicillin G (CAS Registry No.: 61- 02
• Methicillin (CAS Registry No.: 61-32-5); Nafcillin (CAS Registry No.: 147-52-4); Oxacillin (CAS Registry No.: 66- 79-5); Cloxacillin (CAS Registry No.: 61-72-3); Dicloxacillin (CAS Registry No.: 3116-76-5); Ampicillin (CAS Registry No.: 69-53-4); Amoxicillin (CAS Registry No.: 26787-78-0); Ticarcillin (CAS Registry No.
• Anti-fungals agents include but are not limited to Amphotericin B, Candicidin, Dermostatin, Filipin, Fungichromin, Hachimycin, Hamycin, Lucensomycin, Mepartricin, Natamycin, Nystatin, Pecilocin, Perimycin, Azaserine, Griseofulvin, Oligomycins, Neomycin, Pyrrolnitrin, Siccanin, Tubercidin, Viridin, Butenafine, Naftifine, Terbinafine, Bifonazole, Butoconazole, Chlordantoin, Chlormidazole, Cloconazole, Clotrimazole, Econazole, Enilconazole, Fenticonazole, Flutrimazole, Isoconazole, Ketoconazole, Lanoconazole, Miconazole, Omoconazole, Oxiconazole, Sertaconazole, Sulconazole, Tioconazole, Tolciclate, To
• Antiviral agents include, but are not limited to, Acyclovir, Cidofovir, Cytarabine, Dideoxyadenosine, Didanosine, Edoxudine, Famciclovir, Floxuridine, Ganciclovir, Idoxuridine, Inosine Pranobex, Lamivudine, MADU, Penciclovir, Sorivudine, Stavudine, Trifluridine, Valacyclovir, Vidarabine, Zalcitabine, Zidovudine, Acemannan, Acetylleucine, Amantadine, Amidinomycin, Delavirdine, Foscarnet, Indinavir, Interferon- ⁇ , Interferon- ⁇ , Interferon- ⁇ , Kethoxal, Lysozyme, Methisazone, Moroxydine, Nevirapine, Podophyllotoxin, Ribavirin, Rimantadine, Ritonavir2, Saquinavir
• Antiinflammatory agents include, but are not limited to, Enfenamic Acid, Etofenamate, FIufenamic Acid, Isonixin, Meclofenamic Acid, Mefenamic Acid, Niflumic Acid, Talniflumate, Terofenamate, Tolfenamic Acid, Aceclofenac, Acemetacin, Alclofenac, Amfenac, Amtolmetin Guacil, Bromfenac, Bufexamac, Cinmetacin, Clopirac, Diclofenac, Etodolac, Felbinac, Fenclozic Acid, Fentiazac, Glucametacin, Ibufenac, Indomethacin, Isofezolac, Isoxepac, Lonazolac, Metiazinic Acid, Mofezolac, Oxametacine, Pirazolac, Proglumetacin, Sulindac, Tiaramide, Tolmetin, Tropesin, Zomepir
• Beta lactamase inhibitors include, but are not limited to Clavulanic acid, Sulbactam, Sultamacillin, and Tazobactam.
• Compounds of the invention may also be combined with one or more efflux pump inhibitor, such as a quinazolinone efflux pump inhibitors, d-ornithine-d-homophenylalanine-3- aminoquinoline, Phe-Arg-b-naphthylamide, propafenone, a phenothiazine or thioxanthene efflux pump inhibitor, l-aza-9-oxafluorenes, N-[4-[2-(3,4-dihydro-6,7-dimethoxy-2(lH)- isoquinolinyl)ethyl]phenyl]-9, 10-dihydro-5-methoxy-9-oxo-4-Acridinecarboxamide, reserpine, Milbemycin, Cinchonine, Verapamil, L-phenylalanyl-N-2-naphthalenyl-L-Argininamide (and analogs), 5'-methoxyhydnoc
• the compounds of the invention are prepared according to methods well-known to those skilled in the art of organic chemical synthesis.
• the starting materials used in preparing the compounds of the invention are known, made by known methods, or are commercially available.
• the compounds of the invention may have one or more chiral center.
• one optical isomer including diastereomers and enantiomers
• another optical starting materials for example by chiral starting materials, catalysts or solvents
• both stereoisomers or both optical isomers including diastereomers and enantiomers at once (a racemic mixture).
• the compounds of the invention may exist as racemic mixtures, mixtures of optical isomers, including diastereomers and enantiomers, or stereoisomers maybe separated using known methods, such as through the use of, for example, chiral salts and chiral chromatography.
• one optical isomer including a diastereomer and enantiomer, or a stereoisomer
• a racemic mixture is discussed herein, it is clearly contemplated to include both optical isomers, including diastereomers and enantiomers, or one stereoisomer substantially free of the other.
• the invention also includes also includes all energetically accessible conformational and torsional isomers of the compounds disclosed.
• the chemical shifts for 1 H and 13 C are reported in parts per million ( ⁇ ) relative to external tetramethylsilane and are referenced to signals of residual protons in the deuterated solvent.
• the chemical shifts for 19 F are reported in parts per million ( ⁇ ) relative to external fluorotrichloromethane.
• Assignment of NMR data is based on two-dimensional correlation experiments ( 1 H- 1 H COSY, 1 H- 13 C HMQC, 1 H- 13 C HMBC, and 1 H- 1 H NOESY) and the usual principles of NMR spectroscopy (the magnitudes of coupling constants and chemical shifts).
• Analytical HPLC is performed using a YMC Pack Pro C18 50 x 4.6 mm 5 ⁇ m column with an isocratic elution of 0.24 min at 90:10 H 2 O:CH 3 CN containing 0.1% TFA followed by a 4-min linear gradient elution from 90:10 to 10:90 at a flow rate of 2.5 mL/min with UV detection at 254 nm.
• Compound B is prepared using the general method of Wierenga and Skulnick (Wierenga, W.; Skulnick, H. I. J. Org. Chein. (1979) 44: 310-311).
• H-Butyllithium 1.6 M in hexanes
• tetrahydrofliran 10 mL
• ethyl hydrogen malonate 180 juL, 1.50 mmol
• 2,2'-bipyridyl ⁇ 1 mg as indicator.
• the temperature of the reaction mixture is allowed to rise to ca. -5 0 C during the addition of n- butyllithium.
• n-butyllithium (2.8 mL, 4.48 mmol) is added until a pink color persists at -5 0 C for 5-10 min.
• a solution of 2,4,5-trifluoro-3-methoxybenzoyl chloride (0.75 mmol, vide supra) in tetrahydrofuran ( ⁇ 3 mL) is added in one portion to the reaction mixture that had been recooled to -78 0 C.
• the resulting mixture is allowed to warm to room temperature, diluted with ethyl acetate (50 mL), and quenched with a 1 M aqueous solution of hydrochloric acid.
• reaction mixture is diluted with ethyl acetate (100 mL) and quenched by addition of a saturated aqueous solution of ammonium chloride (30 mL).
• the organic layer is washed with brine (4 x 30 mL), dried over sodium sulfate, and evaporated under reduced pressure to give the crude product.
• This material is purified by flash column chromatography (eluting with 40% v/v ethyl acetate in hexanes) to give C as a yellow oil.
• m-Chloroperoxybenzoic acid ( ⁇ 77%, 34 mg, 0.15 mmol) is added in one portion to a solution of D (50 mg, 0.14 mmol) in methylene chloride (3 mL) at room temperature.
• the reaction mixture is stirred for 1 h, diluted with ethyl acetate (20 mL), and washed with a 5% aqueous solution of sodium bicarbonate (2 x 10 mL).
• the organic layer is dried over sodium sulfate and evaporated under reduced pressure to give the crude product.
• Step 6 Synthesis of ethyl l-cyclopropyl- ⁇ J-difluoro-l-mercaptoS-methoxy- ⁇ oxo-lA- dihydroquinoline-3-carboxylate (F).
• Anhydrous sodium hydrogen sulfide (Alfa Aesar, 20 mg, 0.36 mmol) is added in one portion to a solution of DMF (6 mL) containing E (93 mg, 0.24 mmol) at room temperature.
• the resulting solution is heated at 40 0 C for 2-3 h (until TLC indicated complete consumption of E) and allowed to cool to room temperature.
• the reaction mixture is quenched by addition of a 5% aqueous solution of hydrochloric acid (20 mL) and extracted with ethyl acetate (2 x 25 mL).
• a solution of sodium bicarbonate (820 mg, 9.8 mmol) in water (14 mL) is added to a solution of F (348 mg, 0.98 mmol) in tetrahydrofuran (10 mL) at room temperature.
• Hydroxylamine-O-sulfonic acid (465 mg, 4.1 mmol) is added in one portion to this mixture.
• the reaction mixture is stirred at room temperature for ⁇ 3 h and quenched by addition of an aqueous solution of 5% hydrochloric acid (100 mL).
• the precipitate that formed is collected by filtration, washed with water (3 x 5 mL), and dried in vacuo to give 3 as a white solid.
• Compound I is prepared using the general method of Wierenga and Skulnick (Wierenga, W.; Skulnick, H. I. J. Org. Chem. 1979, 44, 310-311).
• n-Butyllithium 1.6 M in hexanes
• tetrahydrofuran 50 mL
• ethyl hydrogen malonate 2.6 mL, 22 mmol
• 2,2'-bipyridyl ⁇ 1 mg as indicator.
• the temperature of the reaction mixture is allowed to rise to ca. -5 °C during the addition of ⁇ -butyllithium.
• Step 6 Synthesis of ethyl l-cyclopropyl-7-fluoro-8 ⁇ methoxy-2 ⁇ (methylsulfinyl)-4-oxo-l,4- dihydroquinoline-3-carboxylate (L).
• Step 7 Synthesis of ethyl l-cyclopropyl-l-fluoro-l-mercaptoS-meihoxy- ⁇ oxo-lA- dihydroquinoline-3-carboxylate (M).
• Anhydrous sodium hydrogen sulfide (Alfa Aesar, 137 mg, 2.45 mmol) is added in one portion to a solution of dimethylformamide (10 mL) containing L (600 mg, 1.63 mmol) at -5 0 C. The resulting mixture is stirred for 15 min (until TLC indicated complete consumption of L) and allowed to warm to room temperature. The reaction mixture is quenched by addition of a 5% aqueous solution of hydrochloric acid (75 mL) and extracted with ethyl acetate (2 x 100 mL).
• a solution of sodium bicarbonate (1.3 g, 15.47 mmol) in water (22 mL) is added to a solution of M (540 mg, 1.60 mmol) in tetrahydrofuran (16 mL) at room temperature.
• Hydroxylamine-O-sulfonic acid (761 mg, 6.73 mmol) is added in one portion to this mixture.
• the reaction mixture is stirred at room temperature for ⁇ 3 h and quenched by addition of an aqueous solution of 5% hydrochloric acid (150 mL).
• the precipitate that formed is collected by filtration, washed with water (3 x 10 mL), and dried in vacuo to give 4 as a white solid.
• Methanesulfonyl chloride (15 mL, 0.19 mol) is added to a cooled (0 0 C) solution of toluene (300 mL) containing (5)-l-benzylpyrrolidin-3-ol (24.5 g, 0.14 mol) and triethylamine (80 mL, 0.57 mol). The resulting mixture is stirred at 0 °C for 15 min, and allowed to warm to room temperature with stirring for 2h. The mixture is quenched with a 5% aqueous solution of sodium bicarbonate (250 mL).
• a mixture containing P (7.4 g), 20% palladium hydroxide on carbon (7.5 g), and ethanol (75 niL) is stirred under an atmosphere of hydrogen gas (50 psi) at 45 °C for 24 h.
• the mixture is filtered and the filtrate is concentrated under reduced pressure to give 5 (4.1 g, 95 %) as a yellow oil.
• This material is stored under an atmosphere of argon gas.
• a reaction vessel is charged with 5 (206.0 mg, 1.6 mmol), 3 (328.6 mg, 1.0 mmol), dimethyl sulfoxide (4.5 mL), and ⁇ N-diisopropylethylamine (750 ⁇ L, 4.3 mmol).
• the resulting mixture is irradiated with microwaves (CEM Discover) at 125 0 C for 1 h (conventional heating may also be used — 115 °C in an oil bath for 14 h), allowed to cool, and evaporated to dryness under reduced pressure (-70 °C/2-3 mm Hg).
• Preparative HPLC is performed using a YMC Pack Pro C18 150 x 30.0 mm 5//m column coupled to a YMC Pack Pro 50 x 20 mm 5/an column with an isocratic elution of 0.37 min at 95:5 H 2 OiCHsCN containing 0.1% TFA followed by a 15.94 min linear gradient elution from 95:5 to 25:75, followed by a 0.69 min linear gradient from 25:75 to 5:95 at a flow rate of 30.0 mL/min with UV detection at 254.
• the crude material is loaded as a solution containing acetic acid ( ⁇ 2 mL), methanol ( ⁇ 1 mL), and water ( ⁇ 1 mL).
• the purified product is isolated as the TFA salt and is converted to the corresponding hydrochloride salt by addition of a solution of hydrogen chloride ( ⁇ 1.25 M in methanol) followed by evaporation; this process is repeated twice to give a yellow solid.
• Purity by HPLC: >99%; t R 10.08 min.
• l-((R)-Pyrrolidin-3-yl)cyclopropanamine (10) is prepared using the method of Inagaki et al. (Inagaki, H.; Miyauchi, S.; Miyauchi, R. N.; Kawato, H. C; Ohki, H.; Matsuhashi, N.; Kawakami, K.; Takahashi, H.; Takemura, M. J. Med. Chem. 2003, 46, 1005-1015).
• the antimicrobial activity of the compounds of the inyention may be evaluated by a number of methods, including the following visual minimum inhibitory concentration (MIC) assay. This assay determines the minimum concentration of compound required to inhibit growth of a bacterial strain.
• MIC visual minimum inhibitory concentration
• test organism ⁇ 1 x 10 6 cfu/mL
• the final test concentrations ranges from 0.125-128 ⁇ g/mL.
• Inoculated plates are incubated in ambient air at 37°C for 18 to 24 hours.
• the organisms selected for testing included laboratory strains S. aureus ATCC 29213 and E. coli ATCC 25922 (strains purchased from American Type Culture Collection, Manassas, VA), S. aureus FQR700699, and Paeruginosa 27853.
• the minimum inhibitory concentration (MIC) is determined as the lowest concentration of compound that inhibited visible growth of the test organism.
• Compounds 17-46, 49, and 52-113 were tested in this assay for anti-microbial activity against S. Aureus.
• Compounds 17-42, 44-46, 49, 52-83, 85-103, and 105-113 exhibited MIC values of 1 microgram/ ml or less against S. Aureus. Certain of these compounds exhibited an MIC value of 0.1 microgram/ ml or less against S. Aureus, and certain of these compounds also exhibited an MIC value of 0.01 microgram/ ml against S. Aureus when tested in this assay.
• Compounds 17, 20, 22, 38-46, 49, and 52-113 were tested in this assay for anti-microbial activity against Methicillin Resistant S. Aureus.
• Optimal cell density is first determined by plating cells in a 96-well plate standard sterile tissue culture plates in 100 ⁇ l media, 10%FBS at six cell densities from 500 cells/ well to 15,000 cells/ well. A cell free well containing only media is used as a control. Cells are incubated at 37 oC in a 5% CO 2 incubator for 24 hours. 10% culture volume (lOul) of Alamar Blue (Biosource, DALl 100, 10OmL) is then added. Cells are incubated at 37 oC in a 5% CO2 incubator and read in a Victor V plate reader, 544nm excitation, 590nm emission, at 3, 4, and 24 hours after the addition of Alamar Blue. The cell number vs.
• Cells are plated at optimal cell density in a standard sterile tissue culture 96 well plate, and incubated at 37 0 C O/N in a 5% CO 2 incubator. 12 to 48 hours post-plating media is removed. The cells are washed 1 or 2 times with IX PBS and replaced with fresh media containing the test compound in 1% DMSO. 24 to 72 hours after addition of compound, the media is removed, and the cells washed 1 to 2 times with IX PBS. Fresh media containing 1/10 volume of Alamar Blue is then added. Plates are incubated 4 hours at 37 oC in a 5% CO2 incubator and read in a Victor V plate reader, 544 nm excitation, 590 nm emission.
• Compounds are diluted to 20 micromolar in 1% DMSO and media and screened in duplicate to obtain single concentration cytotoxicity data. Eight concentration points from 0.78 micromolar to 100 micromolar, run in duplicate, are used to determine cytotoxicity CC50 values. Cells with 1% DMSO and media are used as a negative control, compounds having a known CC50 against a particular cell type are used as positive controls.
• Examples 1 to 8 exhibit CC50 values greater than 10 uM against each of the cell lines listed below.
• Other cell types include but are not limited to Balb/3TC, CEM-SS, HeLa, Hep2, HepG2, HT-29, MRC-5, SK-N-SH, U-87 MG, 293T, and Huh-7. More preferred are compounds with a CC50 value greater than 50 uM. Most preferred are compounds with a CC50 value greater than 100 uM.

Landscapes

• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Health & Medical Sciences (AREA)
• Veterinary Medicine (AREA)
• Life Sciences & Earth Sciences (AREA)
• Public Health (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Oncology (AREA)
• Communicable Diseases (AREA)
• Tropical Medicine & Parasitology (AREA)
• Virology (AREA)
• Epidemiology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
• Medicinal Preparation (AREA)
• Agricultural Chemicals And Associated Chemicals (AREA)

Priority Applications (16)

Applications Claiming Priority (2)

Publications (1)

Family

ID=37106335

Family Applications (1)

Country Status (19)

Cited By (9)

Families Citing this family (10)

Citations (2)

Family Cites Families (23)

• 2006
  • 2006-07-27 KR KR1020087004769A patent/KR101272263B1/ko active Active
  • 2006-07-27 CN CN200680033548XA patent/CN101263146B/zh active Active
  • 2006-07-27 AT AT06800428T patent/ATE482219T1/de active
  • 2006-07-27 BR BRPI0614456A patent/BRPI0614456B8/pt active IP Right Grant
  • 2006-07-27 WO PCT/US2006/029302 patent/WO2007014308A1/en not_active Ceased
  • 2006-07-27 US US11/494,205 patent/US8044204B2/en active Active
  • 2006-07-27 CA CA2616768A patent/CA2616768C/en not_active Expired - Fee Related
  • 2006-07-27 AU AU2006272571A patent/AU2006272571B2/en not_active Ceased
  • 2006-07-27 PL PL06800428T patent/PL1913004T3/pl unknown
  • 2006-07-27 EA EA200800444A patent/EA013244B1/ru unknown
  • 2006-07-27 NZ NZ566209A patent/NZ566209A/en unknown
  • 2006-07-27 DK DK06800428.2T patent/DK1913004T3/da active
  • 2006-07-27 PT PT06800428T patent/PT1913004E/pt unknown
  • 2006-07-27 EP EP06800428A patent/EP1913004B1/en active Active
  • 2006-07-27 ES ES06800428T patent/ES2352799T3/es active Active
  • 2006-07-27 DE DE602006017082T patent/DE602006017082D1/de active Active
  • 2006-07-27 JP JP2008524168A patent/JP5143000B2/ja not_active Expired - Fee Related
• 2008
  • 2008-01-24 IL IL189017A patent/IL189017A/en active IP Right Grant
  • 2008-02-27 NO NO20081005A patent/NO343376B1/no unknown
• 2011
  • 2011-10-24 US US13/279,852 patent/US8946422B2/en active Active

Patent Citations (2)

Also Published As

Similar Documents

Legal Events