HK1208454B - Pyrimidine derivatives for the treatment of bacterial diseases - Google Patents

Description

Pyrimidine derivatives for the treatment of bacterial diseases

The present invention relates to compounds useful for the treatment of bacterial diseases, particularly diseases caused by certain nonmycobacterial Staphylococcus aureus (Staphylococcus aureus). The compounds can be used in any mammal (e.g., human or animal). The invention also relates to novel compounds, compositions, methods and uses.

Background

Bacterial infections are prevalent worldwide and there is a high demand for compounds to treat bacterial infections. There are several known types/strains of bacteria, and finding compounds that are selectively active against certain types/strains of bacteria is a particular goal in the medical field.

There are several drugs known to have activity against nonmycobacteria, but there is still a need for such compounds, especially since bacteria can acquire resistance to certain compounds/drugs. Compounds having selective activity against certain types/strains of bacteria are clearly advantageous, for example these compounds may have the advantage that the bacteria are unable to establish resistance to bacteria of other strains.

Indeed, it is an object of the present invention to provide compounds having selective activity against specific non-mycobacteria, in particular staphylococcus aureus.

Certain pyrimidine compounds are publicly available or have been published by the chemical abstracts service, but such compounds have not had any particular use attributed to them. Both international patent application WO 2005/070899 and US patent application US 2005/182073 disclose certain pyrimidines that may be used to control harmful organisms, such as organisms that attack plants. International patent application WO 2003/077656 discloses certain pyrimidines useful as antibacterial agents. These documents disclose only certain types of pyrimidines.

US patent US 6887870B 1 discloses various compounds as sodium/proton exchange inhibitors, but does not disclose such compounds for use in the treatment of bacterial infections. International patent applications WO 2011/073378, WO 2011/060976 and WO 2011/061214 clearly disclose certain compounds for use as antibacterial agents, but these documents disclose only a limited range of compounds.

Summary of The Invention

Provided herein are compounds of formula I for use as a medicament/medicament, wherein formula I represents:

wherein:

y represents:

;

(ii) -CF₃；

(iii) -N(C_1-6alkyl radical)₂(e.g., -N (CH)₃)₂)；Or

(iv) C_3-6Cycloalkyl (e.g., cyclopropyl);

N^v、N^w、N^x、N^yand N^zIndependently represent-N = or-C (h) = (or-C (a)⁴) =), but wherein N is^v、N^w、N^x、N^yAnd N^zOnly a maximum of 3 of them can represent-N =;

n represents 0, 1 or 2 (but preferably represents 0);

X¹and X²Independently represents-N-or-C (H) -;

when X is present¹represents-N-time, Q¹Represents a direct bond, -C (O) -or-S (O)₂-；

When X is present¹When represents-C (H) -, Q¹Represents a direct bond or-N (R)^z)-；

R^zRepresents hydrogen or C_1-6An alkyl group;

R^xis represented by C_1-6Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent(s), aryl or heteroaryl (wherein the latter two groups are each independently optionally substituted with one or more groups selected from A)²And A³Substituted with a substituent of (a);

R^y、R^y1and R^y2Independently represent hydrogen, halogen, -CN, -OR¹⁰、-N(R¹¹)(R¹²) Or C_1-6Alkyl (optionally substituted with one or more halogen (e.g., fluorine) atoms);

A¹、A²、A³and A⁴Independently represent halogen, -CN, -OR¹、-S(O)_0-2C_1-3Alkyl radical, C_1-6Alkyl (optionally substituted with one or more halo substituents), heterocycloalkyl (optionally substituted with one or more substituents selected from C_1-3Alkyl and halogen substituents), aryl or heteroaryl (wherein the latter two groups are each optionally substituted)Is one or more selected from B¹And B²Substituted with a substituent of (a);

each R¹And R¹⁰Independently represent hydrogen, C_1-6Alkyl (optionally substituted with one or more halo substituents), aryl or heteroaryl (wherein the latter two groups are optionally substituted with one or more substituents selected from halo, C_1-3Alkyl and-O-C_1-3Alkyl substituent substitution);

R¹¹and R¹²Independently represent hydrogen or C_1-6An alkyl group;

B¹and B²Independently represent halogen (such as chlorine or fluorine), -CN, C_1-6Alkyl (optionally substituted by one or more halogen (e.g. fluorine) atoms), -OH or-O-C_1-6Alkyl (optionally substituted with one or more halogen (e.g., fluorine) atoms),

or a pharmaceutically acceptable salt thereof.

The compounds of formula I above, which are useful as medicaments, may be referred to herein as "compounds of the invention".

Compounds of the invention that may be mentioned include those as defined previously, but:

(a) with the proviso that the compound is not:

(ii) a Or

(b) Wherein Y represents:

;

(ii) -CF₃(ii) a Or

(iii) C_3-6Cycloalkyl (e.g., cyclopropyl).

Pharmaceutically acceptable salts include acid addition salts and base addition salts. Such salts may be formed in conventional manner, for example by reacting the free acid or free base form of a compound of formula I with one or more equivalents of a suitable acid or base, optionally in a solvent, or in a vehicle in which the salt is insoluble, followed by removal of the solvent or the vehicle using standard techniques (e.g. vacuum, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter ion of a compound of the invention in salt form with another counter ion, for example using a suitable ion exchange resin.

For the purposes of this invention, solvates, prodrugs, N-oxides, and stereoisomers of the compounds of the invention are also included within the scope of the invention.

The term "prodrug" of a related compound of the invention includes any compound that, following oral or parenteral administration, is metabolized in vivo to form an experimentally detectable amount of the compound within a predetermined time period, e.g., within an administration time interval of between 6 and 24 hours (i.e., 1-4 times per day). For the avoidance of doubt, the term "parenteral" administration includes all forms of administration other than oral administration.

Prodrugs of the compounds of the present invention may be prepared by modifying functional groups present on the compounds in such a way that, upon administration of such prodrugs to a mammalian subject, the modifications are cleaved in vivo. Such modifications are typically effected by synthesizing the parent compound with prodrug substituents. Prodrugs include compounds of the present invention wherein a hydroxy, amino, mercapto, carboxyl or carbonyl group in a compound of the present invention is bonded to any group that can be cleaved in vivo to regenerate the free hydroxy, amino, mercapto, carboxyl or carbonyl group, respectively.

Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxyl functional groups, ester groups of carboxyl functional groups, N-acyl derivatives, and N-Mannich bases. General information on Prodrugs can be found, for example, in Bundegaard, H. "Design of Prodrugs" (pages l-92), Elesevier, New York-Oxford (1985).

The compounds of the invention may contain double bonds and may therefore exist as E (opposite side (entgegen)) and Z (on the same side (zusammen)) geometric isomers of each individual double bond. Positional isomers are also encompassed by the compounds of the present invention. All such isomers (e.g., cis-and trans-forms are included if the compounds of the present invention incorporate double or fused rings) and mixtures thereof are included within the scope of the present invention (e.g., single positional isomers and mixtures of positional isomers may be included within the scope of the present invention).

The compounds of the invention may also exhibit tautomerism. All tautomeric forms (or tautomers) and mixtures thereof are included within the scope of the invention. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can be interconverted via a low energy barrier. For example, proton tautomers (also referred to as prototropic tautomers) include interchange phenomena via proton migration, such as isomerization of keto-enols and imine-enamines. Valence tautomers include the interchange phenomenon by recombination of certain bonding electrons.

The compounds of the invention may also contain one or more asymmetric carbon atoms and may therefore exhibit optical and/or diastereoisomerism. The diastereomers may be separated using conventional techniques, such as chromatography or fractional crystallization. The various stereoisomers may be isolated by separation of racemates or other mixtures of the compounds using conventional (e.g. fractional crystallisation or HPLC) techniques. Alternatively, the desired optical isomer may be prepared by: by reacting suitable optically active starting materials under conditions that do not lead to racemization or epimerization (i.e., the "chiral pool" method); by reacting the appropriate starting materials with "chiral auxiliaries", which can subsequently be removed in a suitable stage; by derivatization, e.g., with a homochiral acid (i.e., resolution, including dynamic resolution), followed by separation of the diastereomeric derivatives by conventional means, e.g., chromatography; or by reaction with a suitable chiral reagent or chiral catalyst, all under conditions known to the skilled person.

All stereoisomers (including but not limited to diastereomers, enantiomers and atropisomers) and mixtures thereof (e.g., racemic mixtures) are included within the scope of the present invention.

In the structures shown herein, where no stereochemistry of any particular chiral atom is specified, then all stereoisomers are desired and included as compounds of the present invention. Where stereochemistry is indicated by a solid wedge or dashed line representing a particular configuration, then the stereoisomer is so designated and defined.

The compounds of the present invention may exist in unsolvated forms as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the present invention encompass both solvated and unsolvated forms.

The present invention also encompasses isotopically-labelled compounds of the present invention, which are identical to those recited herein, except for the fact that: one or more atoms are replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (or the most abundant atom found in nature). All isotopes of any particular atom or element as specified herein are included within the scope of the compounds of the present invention. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, for example²H、³H、¹¹C、¹³C、¹⁴C 、¹³N、¹⁵O、¹⁷O、¹⁸O、³²P、³³P、³⁵S、¹⁸F、³⁶Cl、¹²³I and¹²⁵I. certain isotopically-labeled compounds of the present invention (for example,³h and¹⁴c-labeled ones) were used for compound and substrate tissue distribution analysis. Tritium (A)³H) And carbon-l 4 (¹⁴C) Isotopes are useful for their ease of preparation and detectability. In addition, the heavy isotopes such as deuterium (i.e.,²H) alternative provisioning factorsCertain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) may therefore be preferred in certain circumstances. Positron emitting isotopes such as¹⁵O、¹³N、¹¹C and¹⁸f was used in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of the present invention can generally be prepared by following procedures analogous to those disclosed in scheme 1 and/or in the examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

Unless otherwise indicated, C is defined herein_1-qAlkyl (where q is the upper limit of the range) may be straight chain, or may be branched and/or cyclic (so forming C) when a sufficient number (i.e. a minimum of two or three, as appropriate) of carbon atoms are present_3-q-cycloalkyl). Such cycloalkyl groups may be monocyclic or bicyclic, and may further be bridged. Furthermore, such groups may also be partially bicyclic when a sufficient number (i.e. a minimum of 4) of carbon atoms are present. Such alkyl groups may also be saturated or, when a sufficient number (i.e., a minimum of two) of carbon atoms are present, unsaturated (e.g., to form C)_2-qAlkenyl or C_2-qAlkynyl).

C which may be mentioned in particular_3-qCycloalkyl (where q is the upper limit of the range) may be monocyclic or bicyclic alkyl, which may be further bridged (e.g., so as to form a fused ring system such as 3 fused cycloalkyl groups). Such cycloalkyl groups may be saturated or unsaturated (containing one or more double bonds) (forming, for example, cycloalkenyl groups). The substituents may be attached to any point of the cycloalkyl group. In addition, such cycloalkyl groups may also be partially cyclic when a sufficient number (i.e., a minimum of 4) carbon atoms are present.

The term "halogen", as used herein, preferably includes fluorine, chlorine, bromine and iodine.

Heterocycloalkyl radicals which may be mentioned include non-aromatic monocyclic and bicyclic heterocycloalkyl radicals in which at least one is present in the ring systemAnd wherein the total number of atoms in the ring system is between 3 and 20 (e.g. between 3 and 10, such as between 3 and 8, for example 5-to 8-). Such heterocycloalkyl groups may also be bridged. Furthermore, such heterocycloalkyl groups may be saturated or unsaturated (containing one or more double and/or triple bonds), forming, for example, C_2-qHeterocycloalkenyl (where q is the upper limit of the range) group. There may be mentioned C_2-qHeterocycloalkyl radicals include 7-azabicyclo [2.2.1]Heptyl, 6-azabicyclo [3.1.1]Heptyl, 6-azabicyclo [3.2.1]-octyl, 8-azabicyclo- [3.2.1]Octyl, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridinyl, dihydropyrrolyl (including 2, 5-dihydropyrrolyl), dioxolanyl (including 1, 3-dioxolanyl), dioxanyl (including 1, 3-dioxanyl and 1, 4-dioxanyl), dithianyl (including 1, 4-dithianyl), dithiolanyl (including 1, 3-dithiolanyl), imidazolidinyl, imidazolinyl, morpholinyl, 7-oxabicyclo [2.2.1 ] 1]Heptyl, 6-oxabicyclo- [3.2.1]Octyl, oxetanyl, piperazinyl, piperidinyl, non-aromatic pyranyl, pyrazolidinyl, pyrrolidinonyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, sulfolane, 3-dioxythienylenyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydropyridinyl (e.g., 1,2,3, 4-tetrahydropyridinyl and 1,2,3, 6-tetrahydropyridinyl), thietanyl, thiiranyl, thietanyl, thiomorpholinyl, trithianyl (including 1,3, 5-trithianyl), tropanyl, and the like. Where appropriate, substituents on heterocycloalkyl groups may be located at any atom in the ring system, including heteroatoms. The point of attachment of the heterocycloalkyl group may be via any atom in the ring system, including (where appropriate) a heteroatom (e.g., a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heterocycloalkyl groups may also be present in the oxidized form of N-or S-. The heterocycloalkyl groups referred to herein may be specified as a particular monocyclic or bicyclic ring.

Aryl radicals which may be mentioned include C_6-20E.g. C_6-12(e.g. C)_6-10) And (4) an aryl group. Such groups may be monocyclic, bicyclic or tricyclic and haveBetween 6 and 12 (e.g., 6 and 10) ring carbon atoms, at least one of which is aromatic. C_6-10Aryl groups include phenyl, naphthyl, and the like, such as 1,2,3, 4-tetrahydronaphthyl. The point of attachment of the aryl group may be via any atom in the ring system. For example, when the aryl group is polycyclic, the point of attachment can be via an atom that includes an atom of a non-aromatic ring. However, when the aryl groups are polycyclic (e.g. bicyclic or tricyclic), they are preferably attached to the remainder of the molecule via an aromatic ring.

The term "heteroaryl" as used herein, unless otherwise indicated, refers to an aromatic group containing one or more heteroatoms (e.g., 1-4 heteroatoms) preferably selected from N, O and S. Heteroaryl groups include those having from 5 to 20 members (e.g., 5 to 10) and can be monocyclic, bicyclic, or tricyclic, provided that at least one ring is aromatic (thus forming, for example, a mono-, bi-, or tricyclic heteroaromatic group). When the heteroaryl is polycyclic, the point of attachment can be via any atom, including atoms of a non-aromatic ring. However, when the heteroaryl groups are polycyclic (e.g., bicyclic or tricyclic), they are preferably attached to the remainder of the molecule via an aromatic ring. Heteroaryl groups which may be mentioned include 3, 4-dihydro-1H-isoquinolinyl, 1, 3-dihydroisoindolyl (e.g.3, 4-dihydro-1H-isoquinolin-2-yl, 1, 3-dihydroisoindol-2-yl; i.e.heteroaryl groups which are linked via a non-aromatic ring), or preferably acridinyl, benzimidazolyl, benzodioxanyl, benzodioxepinyl, benzodioxolyl (including 1, 3-benzodioxolyl), benzofuranyl, benzofurazanyl, benzothiadiazolyl (including 2,1, 3-benzothiadiazolyl), benzothiazolyl, benzoxadiazolyl (including 2,1, 3-benzoxadiazolyl), benzoxazinyl (including 3, 4-dihydro-2H-1, 4-benzoxazinyl), benzoxazolyl, benzomorpholinyl, benzoselenadiazolyl (including 2,1, 3-benzoselenadiazolyl), benzothienyl, carbazolyl, chromanyl, cinnolinyl, furyl, imidazolyl, imidazo [1,2-a ] pyridyl, indazolyl, indolinyl, indolyl, isobenzofuranyl, isobenzodihydropyranyl, isoindolyl, isoquinolyl, isothiazolyl, isothiochromanyl, isoxazolyl, naphthyridinyl (including 1, 6-naphthyridinyl or, preferably, 1, 5-naphthyridinyl and 1, 8-naphthyridinyl), oxadiazolyl (including 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl and 1,3, 4-oxadiazolyl), oxazolyl, phenazinyl, phenothiazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalyl, tetrahydroisoquinolinyl (including 1,2,3, 4-tetrahydroisoquinolinyl and 5,6,7, 8-tetrahydroisoquinolinyl), tetrahydroquinolinyl (including 1,2,3, 4-tetrahydroquinolinyl and 5,6,7, 8-tetrahydroquinolinyl), tetrazolyl, thiadiazolyl (including 1,2, 3-thiadiazolyl, 1,2, 4-thiadiazolyl and 1,3, 4-thiadiazolyl), thiazolyl, thiochromanyl, thiopheneneyl, thienyl, triazolyl (including 1,2, 3-triazolyl, 1,2, 4-triazolyl, and 1,3, 4-triazolyl), and the like. Where appropriate, substituents on heteroaryl groups may be located on any atom in the ring system, including heteroatoms. The point of attachment of the heteroaryl group may be via any atom in the ring system, including (where appropriate) a heteroatom (e.g., a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heteroaryl groups may also be present in the oxidized form of N-or S-. The heteroaryl groups referred to herein may be specified as particular monocyclic or bicyclic rings. When the heteroaryl is polycyclic, in which non-aromatic rings are present, then the non-aromatic rings may be substituted with one or more = O groups.

It is expressly intended that the heteroaryl group is monocyclic or bicyclic. In the case where a given heteroaryl is bicyclic, then it may consist of a 5-, 6-or 7-membered monocyclic (e.g., monocyclic heteroaryl ring) fused to another 5-, 6-or 7-membered ring (e.g., monocyclic aryl or heteroaryl ring).

Heteroatoms which may be mentioned include phosphorus, silicon, boron and preferably oxygen, nitrogen and sulfur.

For the avoidance of doubt, groups (e.g. C) are specified herein_1-6Alkyl) may be substituted by one or more substituents (e.g. selected from A)¹) When substituted, those substituents (e.g. from A)¹Definitions) are independent of each other. That is toSuch groups may be substituted with the same substituents (e.g. by A)¹Definitions) or different substituents (from A)¹Definition).

All of the individual features mentioned herein (such as the preferred features) may be employed alone or in combination with any of the other features mentioned herein (including the preferred features) (accordingly, the preferred features may be employed in combination with, or independently of, the other preferred features).

The skilled person will appreciate that the compounds of the invention which are the subject of the present invention include those which are stable. That is, the compounds of the present invention include those that are robust enough to survive isolation to a useful degree of purity from, for example, a reaction mixture.

Compounds of the invention that may be mentioned include those in which a compound of formula I as defined herein is provided, but:

provided that when Y represents 2-chloro-phenyl, R^y1represents-OCH₂CH₃，R^yAnd R^y2Both represent hydrogen, X¹And X²Both represent N, Q¹When represents a direct bond, then R^xDoes not represent-C (O) O-tert-butyl.

Preferred compounds of the present invention will now be described.

Preferred compounds of the invention include the following compounds, wherein:

-Q¹-R^xdoes not represent-CH₃；

For example, when X represents-N-, and Q¹Represents a direct bond, and R^xWhen it represents an alkyl group, then it preferably represents C_2-6(e.g. C)_3-6) Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent of (a);

when X represents-N-and Q¹When represents a direct bond, then R^xPreferably represents C_2-6(e.g. C)_3-6) Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent(s), aryl or heteroaryl (wherein the latter two groups are each optionally substituted with one or more groups selected from A)²And A³Substituted with a substituent of (a);

when R is^xWhen it represents an alkyl group, then it preferably represents C_2-6(e.g. C)_3-6) Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent of (a).

Preferred compounds of the invention include the following compounds, wherein:

the substructure of formula I below:

is a substructure in which, preferably:

N^v、N^w、N^x、N^yand N^zNone, one or two (preferably one, N)^xOr N^y) represents-N = and the others represent-c (h) =, or N in case the above ring represents phenyl^v、N^w、N^x、N^yAnd N^zOne of them represents-C (A)⁴)=；

When N is present^v、N^w、N^x、N^yAnd N^zis-N = then it is preferably N^wAnd N^y(so formed 5-pyrimidinyl);

n represents 0, 1 or 2 (but preferably represents 0);

A⁴(which may be present on any carbon atom of the aromatic ring, including on N^x/N^y/N^zWhen represents-C (H) = halogen (e.g. fluorine or bromine), -CN, -OC_1-3Alkyl (e.g. -OCH)₃)、-S(O)₂C_1-3Alkyl or C_1-3Alkyl (optionally containing unsaturation, thus forming, for example, -C.ident.C), although A⁴Preferably absent;

more preferably, the above substructure represents pyrimidinyl or pyridinyl (preferably pyridinyl), optionally substituted by one or more groups selected from a⁴Substituted with the substituent(s);

most preferably, the above substructure represents a pyridyl group (preferably an unsubstituted pyridyl group, such as a 2-pyridyl group, or preferably, a 3-pyridyl or 4-pyridyl group) or a substituted phenyl group.

In one embodiment of the invention:

y represents a group as defined herein containing N^vTo N^zIs preferred (this is most preferred).

In another embodiment of the invention:

y represents-N (C)_1-6Alkyl radical)₂(e.g., -N (CH)₃)₂)。

In another embodiment of the invention:

y represents C_3-6Cycloalkyl (e.g., cyclopropyl).

In another embodiment of the invention:

y represents-CF₃。

More preferred compounds of the invention include the following compounds, wherein:

when X is present¹represents-N-time, Q¹Represents a direct bond;

X²denotes-C (H) -and X¹represents-N- (so forming a 4-piperidinyl group);

X²represents-N-and X¹Represents-c (h) - (thus forming a 1-piperidinyl group);

X¹and X²represents-N-, thus forming a piperazinyl group.

More preferred compounds of the invention include the following compounds, wherein:

A¹represents heterocycloalkyl (e.g. oxetanyl) or, more preferably, A¹Represents halogen (such as fluorine), -CN, C_1-6Alkyl (e.g. C)_3-6Cycloalkyl), aryl (optionally substituted by one or more (e.g. one) selected from B¹Substituted with one or more (e.g. one) substituents selected from B), heteroaryl (optionally substituted with one or more (e.g. one) substituents selected from B)²Substituted with a substituent) OR-OR¹；

When A is¹When aryl is represented, it is preferably optionally selected from B by one or more (e.g. one or two)¹Phenyl substituted with the substituent of (1);

when A is¹The representation being represented by one or more (e.g. one) B¹When the substituent is substituted for aryl, then at least one substituent located meta to the phenyl group (and in general, preferably one or two B's are present)¹A substituent);

when A is¹When represents optionally substituted heteroaryl, it is preferably a 5-or 6-membered heteroaryl, preferably containing one, two or three (e.g. one) heteroatoms preferably selected from nitrogen, sulphur and oxygen (e.g. sulphur and/or oxygen), thus forming for example a thienyl (e.g. 2-thienyl or 3-thienyl) or furyl (e.g. 2-furyl) group;

when A is¹When an optionally substituted heteroaryl group is represented, then it is optionally substituted by one or two (e.g. one) groups selected from B²Substituted with the substituent(s);

when A is¹Is represented by C_3-6When cycloalkyl, then it is preferably cyclohexyl;

B¹represents halogen (such as fluorine or chlorine), -CN, -OH or C_1-3Alkyl (methyl; optionally substituted by one or more halogen, e.g. fluorine atoms, thus forming, for example, -CF₃A group);

B²preferably represents C_1-4Alkyl (e.g. C)_1-2Alkyl groups such as methyl);

A¹denotes halogen (e.g. fluorine), -CN, thienyl (e.g. 2-or 3-thienyl, e.g. 3-methyl, 2-thienyl or unsubstituted 3-thienyl), furyl (e.g. 2-furyl), C_3-6Cycloalkyl (such as cyclohexyl) or-O-phenyl;

R¹represents an aryl group (e.g., unsubstituted phenyl);

when Q is¹represents-N (R)^z) When then R^xIs represented by C_1-6Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent(s) so as to form, for example, -C (O) -C (H) (CH)₃) -O-phenyl;

R^zrepresents hydrogen;

when Q is¹Represents a direct bond or-C (O) -then R^xPreferably represents:

C_1-6alkyl (e.g. acyclic C)_1-6Alkyl or C_3-6Cycloalkyl), optionally substituted with one or more substituents selected from = O and/or a¹And optionally contains one or more (e.g. one) double bonds (thus forming C)_2-6Alkenyl) or triple bond (so formed C)_2-6Alkynyl) so as to form, for example, -CH₂-C(CH₃)₃、-CH₂CH(CH₃)₂Cyclopropyl, -CH₂-CF₃、-CH₂-C(H)F₂、-CH₂C(CH₃)₂-CN、-C(O)-C(CH₃)₃、-CH₂-CF₂CH₃、-CH₂- [ 3-methyl, 2-thienyl ] group]、-CH₂-C(CH₃)=CHCH₃、-CH₂- [ 3-fluorophenyl group]、-CH₂- [ 3-thienyl ] group]、-CH₂- [ 3-chloro-6-OH-phenyl]、-CH₂- [ 3-hydroxyphenyl group]、-CH₂- [ 2-hydroxyphenyl group]、-CH₂- [ 2-hydroxy-4-chloro-phenyl group]、-CH₂- [ 2-hydroxy-5-chlorophenyl ] -carboxylic acid]、-CH₂-phenyl, -CH₂-cyclohexyl, -CH₂- [ 2-thienyl ] group]、-CH₂- [ 2-furyl group]、-C(O)-C(H)(CH₃) -O-phenyl, -CH₂-C(H)(CH₃)₂-cyclopropyl, -CH₂- [ 4-fluorophenyl group]、-C(O)-C(CH₃)₃、-CH₂- (3-trifluoromethyl-phenyl), -CH₂- (3-cyanophenyl), -CH₂- (4-cyanophenyl), -CH₂- (2, 4-difluorophenyl), -CH₂- (3-methylphenyl), -CH₂- (4-methylphenyl), -CH₂- (2-fluorophenyl), -CH₂- (2-cyanophenyl), -CH₂- (3, 4-difluorophenyl), -CH₂- (4-chlorophenyl), -CH₂- (3-chlorophenyl), -CH₂- (2-trifluoromethyl-phenyl), -CH₂- (2, 6-difluorophenyl), -CH₂- (3, 5-difluorophenyl), -CH₂-C ≡ CH or-CH₂-C(CH₂) (3-Oxetanyl) (most preferably, R^xrepresents-CH₂-C(CH₃)₃、-CH₂-CF₃、-CH₂-C(H)F₂、-CH₂C(CH₃)₂-CN、-C(O)-C(CH₃)₃or-CH₂-CF₂CH₃) (ii) a Or

R^xRepresents optionally one or more (e.g. one or two) groups selected from A²Aryl (e.g., phenyl) substituted with the substituents of (a), thus forming, for example, unsubstituted phenyl;

R¹⁰is represented by C_1-4Alkyl (e.g. C)_1-2Alkyl groups such as methyl);

R¹¹and R¹²Independently represent hydrogen, or preferably, C_1-3Alkyl (e.g., methyl);

or R^y、R^y1And R^y2All represent hydrogen, or more preferably R^y、R^y1And R^y2At least one of (preferably R)^y) Represents a substituent other than hydrogen and others (preferably R)^y1And R^y2) Represents hydrogen (i.e., preferably one substituent present on the phenyl ring, preferably in the meta position);

when R is^y、R^y1And R^y2One (such as R)^y) When a substituent is represented, then it is preferably selected from halogen, -OCH₃、-N(CH₃)₂CN or C optionally substituted by one or more fluorine atoms_1-3An alkyl group;

R^yrepresents hydrogen, or preferably, halogen (such as fluorine, or preferably chlorine), -OCH₃、-N(CH₃)₂CN or C optionally substituted by one or more fluorine atoms_1-3Alkyl (e.g. -CH)₃) (e.g., -CF)₃) And most preferably, R^yrepresents-OCH₃or-CN;

R^y1represents hydrogen, or when R^yWhen represents hydrogen, R^y1May represent a group selected from-OCH₃And C_1-3Alkyl (e.g., methyl) substituents;

R^y2represents hydrogen, or when R^yAnd R^y1When represents hydrogen, R^y2May represent a group selected from halogen (e.g. fluorine) and C_1-3Alkyl (e.g., methyl) substituents;

R¹represents hydrogen;

A²and A³Independently represents halogen (e.g. chlorine) OR-OR¹(e.g., -OH).

Certain of the compounds of the invention disclosed herein may be novel per se. A further embodiment of the invention therefore provides a compound of formula I:

but wherein:

y represents:

；

N^v、N^w、N^x、N^yand N^zNone or one of them (preferably one, e.g. N)^xOr N^y) represents-N = and the others represent-c (h) =;

n represents 0 or 1;

X¹and X²Independently represents-N-or-C (H) -;

when X is present¹represents-N-time, Q¹Represents a direct bond;

when X is present¹When represents-C (H) -, Q¹Represents a direct bond or-N (R)^z)-；

R^zRepresents hydrogen or C_1-6An alkyl group;

R^xis represented by C_1-6Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent(s), aryl or heteroaryl (wherein the latter two groups are each independently optionally substituted with one or more groups selected from A)²And A³Substituted with a substituent of (a);

R^y、R^y1and R^y2Independently represent hydrogen, halogen, -CN, -OR¹⁰、-N(R¹¹)(R¹²) Or C_1-6Alkyl (optionally substituted with one or more halogen (e.g., fluorine) atoms);

A¹、A²、A³and A⁴Independently represent halogen, -CN, -OR¹、-S(O)_0-2C_1-3Alkyl radical, C_1-6Alkyl (optionally substituted with one or more halo substituents), heterocycloalkyl (optionally substituted with one or more substituents selected from C_1-3Alkyl and halogen), aryl or heteroaryl (wherein the latter two groups are each optionally substituted by one or more substituents selected from B¹And B²Substituted with a substituent of (a);

each R¹And R¹⁰Independently represent hydrogen, C_1-6Alkyl (optionally substituted with one or more halo substituents), aryl or heteroaryl (wherein the latter two groups are optionally substituted with one or more substituentsFrom halogen, C_1-3Alkyl and-O-C_1-3Alkyl substituent substitution);

R¹¹and R¹²Independently represent hydrogen or C_1-6An alkyl group;

B¹and B²Independently represent halogen (such as chlorine or fluorine), -CN, C_1-6Alkyl (optionally substituted by one or more halogen (e.g. fluorine) atoms), -OH or-O-C_1-6Alkyl (optionally substituted with one or more halogen (e.g., fluorine) atoms),

or a pharmaceutically acceptable salt thereof, with the proviso that the compound is not:

。

according to this further aspect of the invention, preferred novel compounds of the invention may be those mentioned previously herein, but wherein:

X¹represents-N-;

X²represents-C (H) -;

R^xis represented by C_1-6Alkyl (optionally substituted with one or more groups selected from = O and a)¹Substituted with a substituent of (a);

A⁴(which is preferably present on a carbon atom of the phenyl ring and is preferably in the para-position) represents halogen (such as fluorine), -CN or-OC_1-3Alkyl (e.g. -OCH)₃)；

A¹Represents halogen (such as fluorine), -CN, C_1-6Alkyl OR-OR¹；

Or R^y、R^y1And R^y2All represent hydrogen, or more preferably R^y、R^y1And R^y2At least one of (preferably R)^y) Represents a substituent other than hydrogen, and others (preferably R)^y1And R^y2) Represents hydrogen (i.e.,preferably one substituent present on the phenyl ring, preferably in the meta position);

when R is^yWhen not hydrogen, it preferably represents halogen (e.g. chlorine), -OCH₃or-CN; and/or

R¹Represents hydrogen.

In particular, preferred novel compounds of the invention may be the following:

or a pharmaceutically acceptable salt thereof.

Pharmacology of

The compounds according to the invention have surprisingly been shown to be suitable for the treatment of certain non-mycobacterial infections, in particular staphylococcus aureus. They are therefore used as medicaments/pharmaceuticals.

Furthermore, the present invention also relates to the use of a compound of the invention, a pharmaceutically acceptable salt thereof or an N-oxide form thereof, and any pharmaceutical composition thereof as hereinbefore described, for the manufacture of a medicament for the treatment of certain non-mycobacterial (especially s.

Thus, in a further aspect, the present invention provides a method of treating a patient suffering from or at risk of certain nonmycobacterial (especially staphylococcus aureus) infections, comprising administering to said patient a therapeutically effective amount of a compound or pharmaceutical composition according to the invention.

The compounds of the present invention have not only been shown to be suitable for the treatment of certain non-mycobacterial staphylococcus aureus but also show selective activity against them. Thus, where reference is made herein to "treating" a certain non-mycobacterium, it is preferably meant "selective treatment", e.g. that it has activity against that bacterium(s) (s.aureus), but may have no or little (or very little) activity against other bacteria. This may be advantageous because if the compound/drug is selective only against staphylococcus aureus, resistance to other strains cannot be established and the need for unnecessary antibacterial action is prevented.

Bacterial infections treatable by the present compounds include, for example, central nervous system infections; external ear infections; middle ear infections, such as acute otitis media; cranial sinus infection; eye infections; oral infections, such as tooth, gum and mucosal infections; upper respiratory tract infections; lower respiratory tract infections; urogenital infections; gastrointestinal tract infections; gynecological infection; septicemia; bone and joint infections; infections of the skin and skin structures; bacterial endocarditis; burn; antibacterial prevention of surgery; and antibacterial prophylaxis in immunosuppressed patients, such as patients receiving cancer chemotherapy or organ transplant patients.

When used in the upper or lower context, a compound is useful for treating bacterial infections means that the compound is useful for treating certain non-mycobacterial infections, particularly infections with staphylococcus aureus.

The invention also relates to compositions comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound according to the invention. The compounds according to the invention can be formulated in various pharmaceutical forms for administration purposes. As suitable compositions, all compositions which are generally used for systemic administration of drugs can be cited. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions need to be in unit dosage form, which is particularly suitable for administration orally or by parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, and in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions, there may be employed, for example, water, glycols, oils, alcohols and the like; or in the case of powders, pills, capsules and tablets, solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like may be used. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will typically comprise sterile water, at least in large part, although other ingredients may be included, for example to aid solubility. For example, injectable solutions may be prepared in which the carrier comprises saline solution, glucose solution, or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations.

Depending on the mode of administration, the pharmaceutical composition will preferably comprise 0.05 to 99% by weight, more preferably 0.1 to 70% by weight, even more preferably 0.1 to 50% by weight of the active ingredient, and 1 to 99.95% by weight, more preferably 30 to 99.9% by weight, even more preferably 50 to 99.9% by weight of a pharmaceutically acceptable carrier, all percentages being based on the total composition.

The pharmaceutical compositions may additionally contain various other ingredients known in the art, for example, lubricants, stabilizers, buffers, emulsifiers, viscosity modifiers, surfactants, preservatives, flavoring agents, or coloring agents.

It is particularly advantageous to formulate the above pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect, in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.

The daily dosage of a compound according to the invention will of course vary with the compound used, the mode of administration, the treatment desired and the mycosis indicated. In general, however, satisfactory results will be obtained when the compound according to the invention is administered in a daily dose not exceeding 1 gram, such as in the range from 10 to 50 mg/kg body weight.

In view of the fact that the compounds of the present invention have activity against bacterial infections (e.g., of a certain type as defined herein), the present compounds can be combined with other antibacterial agents to be effective against bacterial infections.

Thus, the invention also relates to a combination of: (a) a compound according to the invention, and (b) one or more other antibacterial agents.

The invention also relates to the following combinations for use as a medicament: (a) a compound according to the invention, and (b) one or more other antibacterial agents.

The present invention also relates to the use of a combination or pharmaceutical composition as defined directly above for the treatment (e.g. selective treatment) of a bacterial infection(s) (a certain type of s.

A pharmaceutical composition comprising a pharmaceutically acceptable carrier and as active ingredients a therapeutically effective amount of (a) a compound according to the invention and (b) one or more other antibacterial agents is also encompassed by the present invention, particularly for the treatment of certain bacterial infections as defined herein.

The weight ratio of (a) a compound according to the invention and (b) other antibacterial agents, when administered as a combination, can be determined by one skilled in the art. The ratio and precise dosage and frequency of administration will depend upon the particular compound according to the invention and the other antibacterial agent used, the particular condition being treated, the severity of the condition being treated, the age, weight, sex, diet, number of administrations and general physical condition of the particular patient, the mode of administration and other drugs which the individual may take, as is well known to those skilled in the art. Furthermore, it will be apparent that the effective daily dose may be reduced or increased depending on the response of the subject being treated and/or depending on the evaluation of the clinician prescribing the compounds of the instant invention. The specific weight ratio of the compound of formula (Ia) or (Ib) and the further antibacterial agent of the present invention may range from 1/10 to 10/1, more particularly from 1/5 to 5/1, even more particularly from 1/3 to 3/1.

The compound according to the invention and one or more other antibacterial agents may be combined in a single formulation, or they may be formulated in separate formulations so that they may be administered simultaneously, separately or sequentially. The invention therefore also relates to a product containing (a) a compound according to the invention and (b) one or more other antibacterial agents as a combined preparation for simultaneous, separate or sequential use in the treatment of a bacterial infection.

General preparation method

The compounds according to the invention can generally be prepared by successive steps, each of which is known to the skilled worker and/or described in the following general scheme:

general procedure 1:

general procedure 2:

general procedure 3:

it is within the knowledge of the skilled person to consider to find suitable temperatures, dilutions and reaction times to optimize the above reaction to obtain the desired compound.

The compounds of formula I may be converted to the corresponding N-oxide forms following art-known procedures for converting a trivalent nitrogen to its N-oxide form. The N-oxidation reaction may generally be carried out by reacting the starting material of formula I with a suitable organic or inorganic peroxide. Suitable inorganic peroxides include, for example, hydrogen peroxide, alkali or alkaline earth metal peroxides such as sodium peroxide, potassium peroxide; suitable organic peroxides may comprise peroxy acids such as perbenzoic acid or halogen-substituted perbenzoic acids (e.g. 3-chloroperoxybenzoic acid), peroxy alkanoic acids such as peroxyacetic acid, alkyl hydroperoxides such as t-butyl hydroperoxide. Suitable solvents are, for example, water, lower alcohols such as ethanol and the like, hydrocarbons such as toluene, ketones such as 2-butanone, halogenated hydrocarbons such as dichloromethane, and mixtures of such solvents.

For example, wherein Y represents a group containing N^vTo N^zThe cyclic compounds of formula I can be prepared by the following process:

(i) for X in¹A compound of formula I, representing-N-, reacting a compound of formula II,

wherein N is^v、N^w、N^x、N^y、N^z、X²、R^y、R^y1、R^y2、A⁴And n is as defined hereinbefore, with the following compounds:

(a) a compound of the formula III,

L¹-Q¹-R^xIII

wherein L is¹Represents a suitable leaving group, such as a chloride, bromide, iodide or sulfonate group;

(b) for Q in¹Represents a direct bond and R^xIs represented by-CH₂-R^xxPart of which is connected to Q¹Wherein the groups together represent R^xMoiety) of the formulaA compound of formula I, wherein the compound is shown in the specification,

O=C(H)(R^xx) IV

wherein R is^xxRepresents R^xPart of a moiety (R)^xIs as defined hereinbefore) and the reaction is carried out under reductive amination reaction conditions, for example conditions known to those skilled in the art, for example as a reaction in a "one pot" process, for example in the presence of a selective reducing agent (which reduces the imine intermediate, but not the aldehyde starting material), for example sodium cyanoborohydride or preferably sodium triacetoxyborohydride, for example in the presence of a mild acid (such as acetic acid), in a suitable solvent (such as dichloromethane). Alternative conditions may also be employed, for example, a condensation reaction is first carried out, followed by a reaction in the presence of a reducing agent (which need not be "imine" selective, e.g. sodium borohydride may be used when the reaction is carried out in two steps);

(ii) for pyrimidines in which the desired pyrimidine is attached to X²(wherein X²A compound of formula I, representing-N-), with a compound of formula VI under aromatic nucleophilic substitution reaction conditions, for example as known in the art, for example in the presence of a base (e.g., an organic base, such as a dialkylamine base, for example N, N-diisopropylethylamine),

wherein L is²Represents a suitable leaving group such as halogen (e.g. chloro);

wherein X¹、Q¹And R^xAs defined hereinbefore;

(iii) for the presence of-CH therein₂A partial compound in a suitable reducing agent such as LiAlH₄In the presence of-C(O) -partial reduction of the corresponding compound;

(iv) reacting a compound of formula VII with a compound of formula VIII or a derivative thereof (e.g. a salt, e.g. HCl salt) under reaction conditions promoting cyclisation (e.g. in the presence of a base, e.g. an inorganic base such as tBuOK, and a suitable solvent, e.g. an alcoholic solvent such as ethanol, which reaction may be carried out at elevated temperature),

wherein L is³Represents a suitable leaving group (preferably an amino moiety, e.g. -N (CH)₃)₂) And a whole body (e.g. R)^y、R^y1、R^y2、Q¹、R^x、X¹And X²) As defined hereinbefore;

wherein the whole (e.g. N)^v、N^w、N^x、N^y、N^z、A⁴And n) is as defined hereinbefore;

(v) for compounds containing-C (F)₂Part of the compound, reacting the corresponding compound containing the-C (O) -part by reaction with a suitable "fluoride" reagent (such as diethylaminosulfur trifluoride; e.g. in the presence of a suitable solvent such as dichloromethane).

The compound of formula II may be prepared by reacting a compound of formula IX or a derivative thereof (e.g. a protected derivative, such as protected on the-N (H) -moiety with, for example, a Boc group) with a compound of formula VIII as hereinbefore defined,

wherein the whole body (such as L)³、R^y、R^y1、R^y2、Q¹、R^xAnd X²) As defined hereinbefore.

The compounds of formula V may be prepared according to the procedures described herein.

VII and IX compounds may be prepared by reaction of the corresponding compound of formula X with a compound of formula XI, for example DMF-DMA in reflux in the presence of a suitable solvent such as an aromatic solvent, for example toluene,

wherein X^1arepresents-X¹-Q¹-R^x(in the case of the preparation of a compound of formula VII) or-N (H) - (in the case of the preparation of a compound of formula IX or a protected portion thereof, e.g. -N (Boc) -), and other integers (e.g. X²、R^y、R^y1And R^y2) As defined hereinbefore;

O=C(H)-L³XI

wherein L is³As defined hereinbefore (and especially represents amino, e.g. -N (CH)₃)₂Thus forming, for example, DMF).

Compounds of formula X may be prepared according to the procedures described herein.

It is clear that in the preceding and following reactions, the reaction product can be isolated from the reaction medium and, if necessary, further purified according to methods generally known in the art, such as extraction, crystallization and chromatography. It is also more evident that the reaction products present in more than one enantiomeric form can be separated from their mixtures by known techniques, in particular preparative chromatography, for example preparative HPLC, chiral chromatography. Individual diastereomers or individual enantiomers may also be obtained by supercritical fluid chromatography (SCF).

The starting materials and intermediates are commercially available compounds or compounds which can be prepared according to conventional reaction procedures generally known in the art.

The following examples illustrate the invention without limiting it.

Experimental part

General procedure 1:

1. synthesis of intermediate B-1:

to A-1 (100 g,0.64 mol) and Et at 0 deg.C₃Boc was added to a solution of N (64.37 g,0.64 mol) in THF (1000 mL)₂O (138.82 g,0.64 mol), and the mixture was stirred at room temperature for 16 hours. The reaction mixture is then poured into H₂O (1000 mL) and extracted with EtOAc (500 mL x 3). The combined organic layers were passed over anhydrous Na₂SO₄Dried and concentrated in vacuo to give intermediate B-1 (138.10 g, yield: 84%).

2. Synthesis of intermediate C-1:

to B-1 (138 g, 0.54 mol) 1L THF and 1L H at 0 deg.C₂Adding LiOH.H into O solution₂O (67.51 g, 1.61 mol). After the addition, the mixture was stirred at 25 ℃ for 15 hours. The organic solvent was removed under reduced pressure. The mixture was extracted with EtOAc (500 mL x 3), the aqueous layer was separated and treated with 0.5M aqueous HCl to adjust to pH = 3 and CH₂Cl₂(1L x 3) extracting. The combined organic layers were passed over anhydrous Na₂SO₄Drying and concentration gave intermediate C-1 (80g, 65%) as a white solid.

3. Synthesis of intermediate D-1:

at 0 ℃ and N₂Then, to C-1 (80g, 0.35 mol) in 1L of anhydrous CH₂Cl₂To the stirred solution of (3) was added CDI (62.24 g, 0.38 mol). After the addition, the mixture was stirred at 25 ℃ for 1 hour and gas formation was observed. Et was added₃N (42.37 g, 42 mol), the mixture was stirred at 25 ℃ for 30 min, then O, N-dimethylhydroxylamine hydrochloride (42.54 g, 0.44 mol) was added. After the addition, the mixture was stirred at 25 ℃ for 15 hours. The mixture is treated with water and NaHCO₃The aqueous solution and the aqueous citric acid monohydrate solution. Separating the organic layer over anhydrous Na₂SO₄Drying and concentration gave intermediate D-1 (80g, 95%) as a white solid.

4. Synthesis of intermediate F-1:

at 0 ℃ and N₂Next, to a stirred solution of D-1 (20 g, 73.44 mmol) in 500mL anhydrous THF was added E-1 (350 mL, 88 mmol). After the addition, the mixture was stirred at 0 ℃ for 2 hours and at 15 ℃ for 6 hours. The mixture was then filtered. Dissolving the solid in NH₄Cl (100mL) and extracted with EtOAc (200 mL. times.2). The combined organic layers were washed with brine (200 mL. times.2) over MgSO₄Drying, filtration and concentration gave 16.2 g of intermediate F-1 as a white solid.

5. Synthesis of intermediate G-1:

a stirred solution of F-1 (15 g, 45 mmol) and DMF-DMA (9 mL, 67.48 mol) in 300mL dry toluene was heated at 110 ℃ under N₂Stirred for 4 h. The solvent was then evaporated under reduced pressure to give 12.15G of intermediate G-1.

6. Synthesis of intermediate I-1:

to a stirred solution of G-1 (2.5G, 6.4 mmol) in ethanol (24 mL) at room temperature was added isonicotinamide hydrochloride H-1 (1.5G, 9.65 mmol) followed by potassium tert-butoxide (1.44G, 12.9 mmol).

The reaction mixture was then heated at 80 ℃ for 16 hours. After 100% consumption of G-1 (monitored by LCMS), the reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was then diluted with dichloromethane (150 mL) and treated with water (150 mL). The aqueous crude mixture was extracted with dichloromethane (2X 150 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude compound was then purified using dichloromethane/ethyl acetate: 50/50 to give the desired intermediate I-1 as an off-white solid (2.58 g, 90% yield).

7. Synthesis of intermediate J-1:

to a solution of I-1 (2.8 g, 6.25 mmol) in dichloromethane (31 mL) at room temperature was added trifluoroacetic acid (5.7 mL). The reaction mixture was then stirred at room temperature for 3 hours. After complete consumption of I-1 (monitored by TLC), the reaction mixture was concentrated in vacuo to give a residue, which was dissolved in dichloromethane (100mL) and treated with saturated aqueous potassium carbonate (100 mL). The aqueous crude mixture was extracted with dichloromethane (2X 100 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to afford the desired intermediate J-1 as an off-white solid (2 g, 92%), which was used in the next step without any further purification.

General procedure 2:

1. synthesis of intermediate L-1:

at 0 ℃ and N₂Next, K-1(500 mL, 125 mmol) was added to a stirred solution of D-1 (30 g, 104 mmol) in 500mL anhydrous THF. The mixture was stirred at 15 ℃ for 18 h. Reacting the reaction mixture with NH₄Cl (250 mL) and EtOAc (500 mL). The organic layer was washed with brine, over MgSO₄Dried, filtered and concentrated. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate =20:1) to give 15.12 g of intermediate L-1.

2. Synthesis of intermediate M-1:

mixing L-1 (14.20 g, 37.14 mmol), Zn (CN)₂(6.54 g, 55.72 mmol) and Pd (PPh)₃)₄A mixture of (2.15 g, 1.86 mmol) in DMF (140 mL) was stirred at 100 ℃ for 18 h. After the mixture was cooled to room temperature, NaHCO was added₃Solution (200 mL). The resulting mixture was extracted with EtOAc (200 mL × 2). The combined organic layers were washed with NaHCO₃(100mL), brine (100mL), over MgSO₄Dried, filtered and concentrated. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate =10:1) to give an intermediateForm M-1 (12.04 g) was a white solid.

3. Synthesis of intermediate N-1:

in N₂Next, a stirred solution of M-1 (12.00 g, 36.54 mmol) and DMF-DMA (6.53 g, 58.81 mmol) in 300mL dry toluene was stirred at 110 ℃ for 4 h. The solvent was then evaporated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate = 1:1) to give intermediate N-1 (10.05 g) as a white solid.

4. Synthesis of Compound P-1:

to a stirred solution of N-1 (800 mg, 2.0 mmol) in acetonitrile (8 mL) at room temperature was added isonicotinimidoamide hydrochloride H-1 (657 mg, 4.1 mmol) followed by DBU (0.93 mL, 6.2 mmol.) then the reaction mixture was heated in a sealed tube at 110 ℃ for 16H after complete consumption of N-1 (monitored by LCMS), the reaction mixture was cooled to room temperature and treated with water (30 mL.) the aqueous crude mixture was extracted with dichloromethane (3 × 30 mL). the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo then the crude compound (1.3 g) was purified using dichloromethane/methanol/ammonium hydroxide solution (33% in H)₂O.) 98/2/0.1 on silica gel to give the desired intermediate O-1 as a pale yellow solid (800 mg, 87% yield).

5. Synthesis of intermediate P-1

Trifluoroacetic acid (2.15 mL) was added to a solution of O-1 (800 mg, 1.8 mmol) in dichloromethane (10 mL) at room temperature. The reaction mixture was then stirred at room temperature for 2 hours. After complete consumption of O-1 (monitored by TLC), the reaction mixture was treated with saturated aqueous sodium carbonate (30 mL). The aqueous crude mixture was extracted with dichloromethane (3X 30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to afford the desired intermediate P-1 as a pale yellow solid (695 mg, quantitative yield), which was used in the next step without any further purification.

General procedure 3:

1. synthesis of intermediate R-1

Q-1 (150 g, 1.53 mol) and pyridine tribromide(635 g, 1.99 mol) in CH₂Cl₂The mixture in (2L) was stirred at 20 ℃ for 96 hours. The reaction mixture was then washed with Na₂S₂O₃Aqueous (2X 1L) and brine (1L) over MgSO₄Drying, filtration and concentration gave intermediate R-1 (380 g, 96%) as a yellow solid.

2. Synthesis of intermediate S-1

In N₂Next, R-1 (170 g, 1.08 mol), H-1 (334.00 g, 1.29 mol) and NaHCO were added₃A mixture of (362.45 g, 4.31 mol) in 2.5L anhydrous MeOH was stirred at 80 deg.C for 12 h. Then coolingThe mixture was cooled and filtered, and the filtrate was concentrated. The residue was purified by chromatography on silica gel (CH)₂Cl₂MeOH =10:1) to give intermediate S-1 (170 g) as a brown solid.

3. Synthesis of intermediate U-1

In N₂Next, S-1 (150 g, 595.08 mmol) and T-1 (131.16 g, 892.62 mmol) were placed in 1500mL EtOH and 300mL H₂NaHCO was added to the stirred mixture in O₃(189.21 g, 1.79 mol) and Pd (PPh)₃)₂Cl₂(15 g) In that respect At 60 ℃ and N₂Next, the reaction mixture was stirred for 8 hours. The mixture was then filtered and the solvent was evaporated under reduced pressure. The residue was washed with EtOAc to afford intermediate U-1. The crude compound was used directly in the next step.

4. Synthesis of intermediate V-1

At 0 ℃ and N₂Next, add U-1 (100 g, crude) in 1500mL of anhydrous CH₂Cl₂To the stirred suspension in (1) was added oxalyl chloride (462.77 g, 3.65 mol) dropwise. DMF (53.30 g, 7.29 mol) was then added and the mixture was heated at 15 ℃ under N₂The reaction mixture was stirred for 4 h. The solvent was then evaporated under reduced pressure. The residue was dissolved in EtOAc (1L) and NaHCO₃In aqueous solution (1L). The organic layer was washed with brine, over MgSO₄Dried and concentrated. The residue was purified by chromatography on silica gel (CH)₂Cl₂MeOH =20:1) to give the crude product. The crude product was washed with EtOH to give 9.4 g of intermediate V-1 as a brown solid and used directly in the next step.

Synthesis of final compound 6:

to a solution of J-1 (250 mg, 0.722 mmol) in dichloroethane (10 mL) was added acetic acid (0.124 mL,2.17 mmol) and 2, 2-dimethylpropionaldehyde (0.157 mL, 1.45 mmol). The reaction mixture was then stirred at room temperature for 2 hours. Sodium triacetoxyborohydride (428 mg, 2 mmol) was then added and the mixture was stirred at room temperature for 3 h. To complete the reaction, 2-dimethylpropionaldehyde (0.157 mL, 1.45 mmol) and acetic acid (0.124 mL,2.17 mmol) were added and the mixture was stirred at room temperature for 1h, then sodium triacetoxyborohydride (428 mg, 2 mmol) was added. The reaction mixture was stirred at room temperature for 12 hours, then diluted with dichloromethane and treated with saturated sodium bicarbonate solution. The aqueous layer was extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude compound was then purified using methylene chloride/methanol/ammonium hydroxide solution (33% in H)₂O) 98/2/0.1 on silica gel to give the desired compound 6 as a white solid (156 mg, 52% yield).

Synthesis of final compound 9:

in N₂To a solution of J-1 (0.15 g,0.433 mmol) and 4-chloro-2-hydroxybenzaldehyde (0.068 g,0.433 mmol) in dichloromethane (4 mL) under an atmosphere was added sodium triacetoxyborohydride (0.138 g,0.649 mmol) in one portion. The reaction mixture was stirred at room temperature overnight. The reaction mixture was loaded directly into preparative TLC and washed with [ heptane (1): EtOAc (2)]Elute 4 times. The main band was scraped off and washed with [ EtOAc (9): MeOH (1)]From SiO₂And (4) eluting. Evaporation of the eluent until dryness gave 0.139 g of Compound 9 (66%).

Synthesis of final compound 12:

in N₂To a solution of J-1 (0.1 g, 0.289 mmol) in methanol (ultra dry) (3 mL) and acetic acid (0.1mL) under an atmosphere was added (1-ethoxycyclopropoxy) trimethylsilyl (0.061 mL, 0.303 mmol) in one portion. The reaction mixture was stirred at room temperature for 0.5 h, then sodium cyanoborohydride (0.027 g,0.433 mmol) was added and the reaction mixture was heated to reflux overnight, then allowed to cool to room temperature and stirred for 24 h. The reaction mixture was loaded directly on preparative TLC and treated with [ CH ]₂Cl₂(95) : MeOH (5)]And (4) eluting. The main band was scraped off and washed with [ EtOAc (9): MeOH (1)]From SiO₂And (4) eluting. The eluent was evaporated until dryness to give 0.089 g of final compound 12 (54%).

Synthesis of final compound 20:

j-1 (100 mg, 0.289 mmol), 2-phenoxypropionic acid (62.4 mg, 0.375 mmol), EDCI (83mg, 0.433 mmol), HOBT (58.5 mg, 0.433 mmol) and NEt₃(61 μ L, 0.433 mmol) in CH₂Cl₂The mixture in (5 mL) was stirred at RT overnight. Water was added and the layers decanted. The organic layer was washed with water and MgSO₄Dry, filter and evaporate the solvent. Crude compound is subjected to CH₂Cl₂/MeOH/NH₄Chromatographic purification (15-40 μm, 30g) of OH 97.5/2.5/0.1 on silica gel. Evaporation of the solvent gave final compound 20 (64%).

Synthesis of final compound 21:

intermediate W-1 was synthesized according to the procedure described for intermediate V-1, using (3-methoxyphenyl) boronic acid instead of T-1.

A solution of W-1 (99 mg, 0.333mmol), X-1 (77 mg, 0.399 mmol) and N, N-diisopropylethylamine (0.142 mL, 0.831 mmol) in THF (20 mL) was stirred at reflux overnight. To complete the reaction, X-1 (236 mg, 1.22 mmol) and N, N-diisopropylethylamine (0.63 mL, 3.7 mmol) were added portionwise over two days and the reaction mixture was stirred at reflux. The reaction mixture was returned to RT and the solvent was removed in vacuo. The remaining brown oil (ca. 0.5 g) was dissolved in MeOH/CH₂Cl₂And the solid was filtered off. Preparative TLC (heptane/ether, 4:1 [3X ]], 9:1 [3x]) 80 mg of a colorless oil were obtained. The material was dissolved in DIPE and heptane was added. The solvent was removed in vacuo to give compound 21 as a colorless solid (70 mg, 56%).

Synthesis of final compound 25:

j-1 (100 mg, 0.29 mmol), trimethylacetyl chloride (35.5 μ L, 0.29 mmol), NEt₃(40 μ L, 0.29 mmol) in CH₂Cl₂The solution in (4 mL) was stirred at room temperature overnight. The mixture was poured into NaHCO₃Using CH in combination with aqueous solutions₂Cl₂And (4) extracting. The combined organic layers were dried, filtered and concentrated to give 120 mg. The crude product was purified by column chromatography (normal phase on stable silica (5 μm 150X30.0mm) with mobile phase gradient from 0% NH₄OH, 100% DCM, 0% MeOH to 0.6% NH₄OH, 94% DCM, 6% MeOH). The solid was crystallized from diisopropyl ether and dried under vacuum pressure at 70 ℃ to give compound 25 (81 mg, 65%).

Synthesis of final compound 26:

oxalyl chloride (0.22 mL, 2.55 mmoles) was added under nitrogenTo S-1 in CH₂Cl₂(50 mL). DMF (0.02 mL) was added dropwise (exothermic) and the reaction mixture was stirred at rt for 3 h. The solvent was removed under vacuum. The crude material Y-1 was used directly in the next step.

4-phenylpiperidine (0.089 g, 0.549 mmole) was added to a suspension of Y-1 (0.099 g, 0.366 mmol) in THF (8 mL). Upon addition, the solid dissolved and the color changed from brown-yellow to purple. N, N-diisopropylethylamine (0.188 mL, 1.098 mmol) was added and the reaction mixture was stirred at reflux overnight. Water and EtOAc were added. The aqueous layer was extracted with EtOAc. The combined organic extracts are purified over Na₂SO₄Dried and the solvent removed under vacuum. The crude product was purified by flash Chromatography (CH)₂Cl₂2% MeOH) to give intermediate Z-1 as a yellow oil (66 mg, 46%).

Z-1 (0.066 g, 0.167 mmol), 2-methoxyphenylboronic acid (0.038 g, 0.25 mmol) and sodium carbonate (0.060 g, 0.566 mmol) in DME (8 mL)/H₂The suspension in O (2 mL) was purged with argon for 5 min. trans-BIS (triphenylphosphine) palladium (II) chloride (6 mg, 8.6. mu. mol) was added and the suspension was purged with argon for 5 min. The reaction mixture (suspension) was stirred at 60 ℃ under argon for 2 h. Addition of H₂O and EtOAc. The solid was filtered off. The layers were separated. The aqueous layer was extracted with EtOAc. The combined organic layers were washed with brine and over Na₂SO₄And (5) drying. The solvent was removed under vacuum. Dissolving the raw material in CH₂Cl₂. Water was added and the mixture was stirred vigorously overnight. The layers were separated. CH for aquifer₂Cl₂And (4) extracting. The combined organic extracts are purified over Na₂SO₄And (5) drying. The solvent was removed under vacuum. Reacting the material with CH₂Cl₂Co-evaporated together. Adding Et₂O is added to the yellow oil. The material is cured. The suspension was dissolved in Et₂Stir in O overnight. The solid is filtered off and Et is used₂O and H₂O wash and dry to give final compound 26 (31%).

Synthesis of final compound 49:

mixing J-1 (100 mg, 0.289 mmol) and K₂CO₃(80 mg, 0.57 mmol), propargyl bromide (solution 80% WT in toluene, 39 μ L, 0.35 mmol) in CH₃CN (4 mL) was stirred at room temperature overnight. Addition of H₂O and CH₂Cl₂The organic phase is decanted off over MgSO₄The powder was dried, filtered and the solvent was evaporated. The crude compound is reacted in CH₂Cl₂/MeOH/NH₄Column chromatography purification of a silica gel column (15-40 μm, 30g) in OH 97/3/0.5 in CH₃After crystallization in CN/diisopropyl ether, 20mg of compound 49 (18%) are obtained.

Synthesis of final compound 55:

j-1 (200 mg, 0.58 mmol), trifluoroacetic anhydride (177 μ L, 1.27 mmol), NEt at room temperature₃(642 μ L, 4.62 mmol) in CH₂Cl₂(4 mL) for 12 h. The mixture was poured into NaHCO₃By using CH in combination with the aqueous solution of₂Cl₂And (4) extracting. The combined organic layers were dried, filtered and concentrated to afford 248 mg of intermediate Z-1. The crude compound was used directly in the next step.

In N₂Flowing, Et at-70 ℃₂O (5 mL) was added to AlCl₃(97 mg, 0.73 mmol) and the mixture was then stirred at 0 ℃ for 10 min. Adding LiAlH dropwise at 0 DEG C₄(1.12 mL, 2.24 mmol) and the mixture was stirred at 0 ℃ for 10 min. Z-1 (248 mg, 0.56 mmol) in THF (5 mL) was added dropwise and the mixture stirred at 0 deg.C for 1 h. The reaction was quenched with ice and EtOAc was added. The layers were decanted. The organic layer was washed with water and MgSO₄Dry, filter and evaporate the solvent. The crude product is purified by reaction in CH₂Cl₂/MeOH/NH₄Column chromatography purification on silica gel (15-40 μm, 30g) in OH 98/2/0.1. The compound was then purified by washing with 2-ethylpyridine (ETHYLPYRIDINE) 6 μm 150X21.2mm (mobile phase 92% CO)₂8% MeOH) to yield compound 55 (45 mg, 19%).

Synthesis of final compound 56:

to a solution of J-1 (250 mg, 0.72 mmol) in dichloromethane (3.6 mL) was added 2, 2-difluoroethyl triflate (230 mg, 1.08 mmol) followed by triethylamine (0.36 mL, 2.16 mmol, 3eq) at room temperature. The reaction mixture was then stirred at 50 ℃ for 16 h and treated with water (5 mL). The aqueous crude mixture was extracted with dichloromethane (3X 10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo (300 mg). The crude compound was then purified on silica gel using ethyl acetate (100%) to afford the desired compound 56 as a white solid (200 mg, 67% yield).

Synthesis of final compound 57:

to a solution of J-1 (250 mg, 0.72 mmol) in dichloromethane (3.6 mL) at room temperature was added A-2(247 mg, 1.08 mmol) followed by triethylamine (0.36 mL, 2.16 mmol). The reaction mixture was then stirred at 50 ℃ for 16 h and treated with water (5 mL). The aqueous crude mixture was extracted with dichloromethane (3X 10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo (650 mg). The crude compound was then purified over silica gel using ethyl acetate/dichloromethane: 70/30 to give the desired compound 57 as a white solid (260 mg, 84% yield).

Synthesis of final compound 58:

to a solution of Q-1 (200 mg, 0.58 mmol) in dichloromethane (3 mL) was added 2,2, 2-trifluoroethyl triflate (0.13 mL, 0.88 mmol) followed by triethylamine (0.24 mL, 1.76 mmol) at room temperature. The reaction mixture was then stirred at reflux for 2 hours. After 80% of Q-1 was consumed (monitored by LCMS), the reaction mixture was treated with water (5 mL). The aqueous crude mixture was extracted with dichloromethane (3X 10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo (185 mg). The crude compound was then purified on silica gel using ethyl acetate/petroleum ether 50/50 to give the desired compound 58 as a white solid (100 mg, 40% yield).

Synthesis of final compound 64:

to a stirred solution of N-1 (800 mg, 2.0 mmol) in acetonitrile (8 mL) at room temperature was added 2,2, 2-trifluoroacetimide amide hydrochloride B-2 (620 mg, 4.1 mmol) followed by DBU (0.93 mL, 6.2 mmol). The reaction mixture was then heated in a sealed tube at 110 ℃ for 38 hours. After 54% of N-1 was consumed (monitored by LCMS), the reaction mixture was cooled to room temperature, diluted with dichloromethane (30mL) and treated with water (30 mL). The aqueous crude mixture was extracted with dichloromethane (3X 30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude compound was then purified on silica gel using petroleum ether/ethyl acetate 70/30 to give the desired intermediate C-2 as a light yellow solid (315mg, 35% yield).

To a solution of C-2 (465 mg, 1.08 mmol) in dichloromethane (5 mL) at room temperature was added trifluoroacetic acid (1 mL). The reaction mixture was then stirred at room temperature for 5 hours. After complete consumption of C-2 (monitored by TLC), the reaction mixture was concentrated in vacuo to give a residue, which was dissolved in dichloromethane (30mL) and treated with saturated aqueous potassium carbonate (30 mL). The aqueous crude mixture was extracted with dichloromethane (3X 30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to afford the desired intermediate D-2 as a pale yellow solid (340 mg, 94% yield), which was used in the next step without any further purification.

To a solution of D-2 (340 mg, 1.02 mmol) in dichloroethane (13 mL) was added acetic acid (0.19 mL, 4.59 mmol) followed by 2, 2-dimethylpropionaldehyde (0.33 mL, 3.07 mmol) at room temperature. The reaction mixture was then stirred at room temperature for 5 hours, then sodium triacetoxyborohydride (867 mg, 4.08 mmol) was added. The reaction mixture was stirred at room temperature for 48 hours, then diluted with dichloromethane (30mL) and treated with saturated sodium bicarbonate solution (30 mL). The aqueous layer was extracted with dichloromethane (3 × 40 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo (400 mg). The crude compound was reacted using a dichloromethane/methanol/ammonium hydroxide solution (33% in H)₂O) 99/1/0.1 on silica gel to give the desired compound 64 as a white solid (260 mg, 63% yield).

Synthesis of final compound 70:

to a stirred solution of J-1 (800 mg, 2.3 mmol) in acetonitrile (9.2 mL) and dichloromethane (4.8 mL) at room temperature was added chloroacetone E-2 (0.27 mL, 3.45 mmol) followed by potassium carbonate (0.64 g, 4.6 mmol). The reaction mixture was then heated at reflux for 8 hours. The reaction mixture was cooled to room temperature, diluted with dichloromethane (30mL) and treated with water (30 mL). The aqueous crude mixture was extracted with dichloromethane (2X 30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to afford the desired intermediate F-2 as a red oil (930 mg, 100% yield), which was used in the next step without any further purification.

To a solution of F-2 (930 mg, 2.3 mmol) in dichloromethane (115 mL) at-78 deg.C was added dropwise diethylaminosulfur trifluoride (DAST) (0.57 mL, 6.9 mmol) and the reaction mixture was stirred at room temperature for 16H, the reaction mixture was diluted with dichloromethane (50 mL) and treated with saturated aqueous sodium carbonate (50 mL) at 0 deg.C, the crude aqueous mixture was extracted with dichloromethane (2 × 50 mL), the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo, and the crude compound was first treated with dichloromethane/methanol/ammonium hydroxide solution (33% in H)₂O) 99/1/0.1, followed by another purification using dichloromethane/ethyl acetate 80/20. The residue was finally triturated with pentane to give the desired compound 70 as a brown gummy solid (60 mg, 6%).

Synthesis of final compound 89:

to a mixture of intermediate G-2 (0.15G, 0.38 mmol) in dichloromethane (20 mL) was added triethylamine (0.12G, 1.14 mmol), followed by the addition of compound I-2 (0.053G, 0.38 mmol) at 0 deg.C. At 25 ℃ and N₂Next, the reaction mixture was stirred for 15 hours. The mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate. The aqueous layer was back extracted with dichloromethane. The combined organic layers were washed with brine, dried and evaporated to give intermediate I-2 (0.16 g, 90%).

To a solution of intermediate I-2 (0.16 g, 0.35 mmol) in dichloromethane (15 mL) at 0 deg.C was added m-CPBA (0.066 g, 0.38 mmol) in portions. The mixture was stirred at 15 ℃ for 20 hours. A solid precipitated out and was filtered through a pad of celite, washing with dichloromethane. The filtrate was purified by high performance liquid chromatography to give compound 89 (16 mg, 12%).

Synthesis of Final Compound 91

At room temperature, N₂Next, triethylsilylacetylene (38 mg, 0.27 mmol) was added to compound 90 (0.12 g, 0.18 mmol), Pd (PPh) in a microwave vessel₃)₄(21 mg, 0.018 mmol), triethylamine (0.22 g,2.16 mmol) and copper (I) iodide (3 mg, 0.011 mmol) in DMF (3 mL). The vessel was capped and irradiated at 110 ℃ for 40 minutes. The reaction mixture was concentrated under vacuum and the residue was diluted with ethyl acetate (30mL) and water (10 mL). Separating the organic layer over Na₂SO₄Dried and the solvent removed under reduced pressure. The crude product was dried under vacuum and used directly in the next step. 0.15 g of crude intermediate J-2 was obtained.

Intermediate J2 (crude, 0.18 mmol) in dry THF (35 mL) was added to a solution of tetrabutylammonium fluoride (1M in THF, 7.5 mL). The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and the crude product was directly purified by basic preparative high performance liquid chromatography (column: C18, eluent: CH)₃CN/ H₂O 97/ 3, 0.05%NH₃.H₂O). The desired fractions were collected and the solvent was removed under reduced pressure. The product was dried under vacuum to give compound 91 (10mg, 13%).

Analytical method

All compounds were characterized by LC-MS. The following LC-MS method was used:

general procedure NOVA (for method NOVAx)

HPLC measurements were performed using an HPLC 1100/1200 (Agilent) system containing a quaternary pump (quaternary pump) equipped with a degasser, autosampler, Diode Array Detector (DAD) and a column as specified in the methods below, the column being kept at room temperature. The MS detector (MS-Agilent simple quadrupole) was configured with an electrospray-APCI ionization source. Nitrogen was used as the nebulizer carrier gas. Data acquisition was performed using the Chemstation data system.

Method NOVA1: in addition to the general procedure NOVA: reverse phase HPLC was performed on a Nucleosil C18 column (3 μm, 3X 150 mm) at a flow rate of 0.42 ml/min. Two mobile phases (mobile phase A: Water TFA 0.1%; mobile phase B: 100% acetonitrile) were used to run the following gradient conditions: from 98% A for 3 min, over 12 min to 100% B, 100% B5 min, then back to 98% A in 2 min and rebalance with 98% AAnd (5) balancing for 6 minutes. An injection volume of 2 μ l was used. The capillary voltage was 2 kV, the corona discharge was maintained at 1 muA and the source temperature was maintained at 250 ℃. A variable voltage is used for the fragmentor. Mass spectra were obtained in electrospray ionization and APCI in positive mode, scanning from 100 to 1100 amu.

Method NOVA2: in addition to the general procedure NOVA: reverse phase HPLC was performed on an Agilent Eclipse C18 column (5 μm,4.6 x150 mm) at a flow rate of 1 ml/min. Two mobile phases (mobile phase A: Water TFA 0.1%; mobile phase B: 100% acetonitrile) were used to run the following gradient conditions: from 98% a for 3 minutes, over 12 minutes to 100% B, 100% B for 5 minutes, then back to 98% a over 2 minutes and equilibrate with 98% a for 6 minutes again. An injection volume of 2 μ l was used. The capillary voltage was 2 kV, the corona discharge was maintained at 1 muA and the source temperature was maintained at 250 ℃. A variable voltage is used for the fragmenter. Mass spectra were obtained in electrospray ionization and APCI in positive mode, scanning from 80 to 1000 amu.

Method NOVA3: in addition to the general procedure NOVA: reverse phase HPLC was performed on a Phenomenex Gemini C18 column (3 μm, 3x 30 mm) at a flow rate of 0.7 ml/min. Two mobile phases (mobile phase A: Water TFA 0.1%; mobile phase B: 100% acetonitrile) were used to run the following gradient conditions: from 98% a to 100% B in 2 minutes, 100% B for 0.5 minutes, then back to 98% a in 0.1 minutes and equilibrate with 98% a for an additional 2.4 minutes. An injection volume of 2 μ l was used. The capillary voltage was 2 kV, the corona discharge was maintained at 1 muA and the source temperature was maintained at 250 ℃. A variable voltage is used for the fragmenter. Mass spectra were obtained in electrospray ionization and APCI in positive mode, scanning from 80 to 1000 amu.

General procedure B (for method Bxxxx)

HPLC measurements were performed using an HPLC Alliance 2695 (Waters) system containing a quaternary pump equipped with a degasser, autosampler, Diode Array Detector (DAD), CLND detector (Antek) and a column as specified in the methods below, the column being maintained at 40 ℃. The MS detector (ZQ-Waters simple quadrupole) was configured with an electrospray ionization source. Nitrogen was used as the nebulizer carrier gas. Data acquisition is performed by using a Masslynx-Openlynx data system.

Method B5301: in addition to general procedure B: reverse phase HPLC was performed on an X-terra MS C18 column (3.5 μm, 4.6X 100 mm) at a flow rate of 1.5 ml/min. Two mobile phases (mobile phase a: water with 0.1% formic acid: 95/methanol: 5%; mobile phase B: 100% methanol) were used to run the following gradient conditions: from 100% A to 5% A/95% B in 12 minutes, and back to 100% A in 1 minute. An injection volume of 10 μ l was used. The cone voltage for both positive and negative ionization was 30V. Mass spectra were obtained in electrospray ionization and APCI in positive mode, scanning from 100 to 1500 amu.

Method B5501: in addition to general procedure B: reverse phase HPLC was performed on a BEH C18 column (1.7 μm, 2.1 x50 mm) at a flow rate of 0.7 ml/min. Two mobile phases (mobile phase A: methanol, B: ammonium acetate in water 10mM: 90%/acetonitrile: 10%) were used to run the following gradient conditions: from 5% A/95% B to 95% A/5% B in 1.3 minutes for 0.2 minutes, and back to 5% A/95% B in 0.2 minutes for 0.3 minutes. An injection volume of 0.75 ml was used. The cone voltage for both positive and negative ionization was 30V. Mass spectra were obtained by electrospray ionization, scanning from 160 to 1000 amu.

General procedure VDR2 (for method V300xV30xx)

LC measurements were performed using a UPLC (high performance liquid chromatography) acquity (waters) system containing a binary pump equipped with a degasser, autosampler, Diode Array Detector (DAD) and a column as specified in the following methods, the column being maintained at a temperature of 40 ℃. The flow is directed from the column to the MS detector. The MS detector was configured with an electrospray ionization source. The capillary needle pressure was 3kV and the source temperature was maintained at 130 ℃ on a Quattro (triple quadrupole mass spectrometer from Waters). Nitrogen was used as the nebulizer carrier gas. Data acquisition was performed using a Waters-Micromass Masslynx-Openlynx data system.

Method V3007V3001: in addition to the general procedure VDR 2: reversed-phase UPLC at Waters Acquity BEH (bridging ethyl)Siloxane/silica hybrid) C18 column (1.7 μm, 2.1 x100 mm) at a flow rate of 0.35 ml/min. Two mobile phases (mobile phase a: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were employed to run the following gradient conditions: from 90% a and 10% B (hold 0.5 min) to 8% a and 92% B in 3.5 min, hold for 2 min and return to the initial condition in 0.5 min, hold for 1.5 min. An injection volume of 2 ml was used. The cone voltage for the positive and negative ionization modes is 20V. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 seconds, using an interscan delay of 0.1 seconds.

General procedure Wuxi (for method WUXIx)

HPLC measurements were performed using an HPLC 1100/1200 (Agilent) system containing a quaternary pump equipped with a degasser, autosampler, Diode Array Detector (DAD) and a column as specified in the following methods, the column being maintained at 50 ℃. The MS detector (Agilent G1946C or 6110) was equipped with an electrospray or APCI ionization source. Nitrogen was used as the nebulizer carrier gas. Data acquisition was performed using an Agilent Chemstation data system.

Method WUXI1: in addition to the general program WUXI: reverse phase HPLC was performed on a YMC-PACK ODS-AQ C18 column (5 μm, 2X 50 mm) at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: water with 0.1% trifluoroacetic acid; mobile phase B: acetonitrile with 0.05% trifluoroacetic acid) were used to run the following gradient conditions: starting at 100% A, hold for 1 minute, in 4 minutes to 40% A/60% B, hold for 2.5 min, and then return to 100% in 0.5 min. An injection volume of 2 μ l was used. The capillary voltage was 2.5 kV for the positive ionization mode and 3kV for the negative ionization mode, and the corona discharge was kept at 4 μ Α if APCI and source temperature were maintained at 200 ℃. The fragmentation voltage was 70V. Mass spectra were obtained by scanning from 100 to 1000 amu in electrospray ionization or APCI in positive mode.

Method WUXI2: in addition to the general program WUXI: reverse phase HPLC was performed on a YMC-PACK ODS-AQ C18 column (5 μm, 2X 50 mm) at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: containing 0.1% trifluoroacetic acid) were usedWater of (2); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid) to run the following gradient conditions: starting at 90% A/10% B, hold for 0.8 min, in 3.7 min to 20% A/80% B, hold for 3 min, and then return to the initial conditions in 0.5 min. The capillary voltage was 2.5 kV for the positive ionization mode and 3kV for the negative ionization mode, and the corona discharge was kept at 4 μ Α if APCI and source temperature were maintained at 200 ℃. The fragmentation voltage was 70V. Mass spectra were obtained by electrospray ionization or APCI in positive mode, scanning from 100 to 1000 amu.

Method WUXI3: in addition to the general program WUXI: reverse phase HPLC was performed on a YMC-PACK ODS-AQ C18 column (5 μm, 2X 50 mm) at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: water with 0.1% trifluoroacetic acid; mobile phase B: acetonitrile with 0.05% trifluoroacetic acid) were used to run the following gradient conditions: starting at 70% A/30% B, hold for 0.8 min, in 3.2 min to 10% A/90% B, hold for 3.5 min, and then return to the initial condition in 0.5 min. The capillary voltage was 2.5 kV for the positive ionization mode and 3kV for the negative ionization mode, and the corona discharge was kept at 4 μ Α if APCI and source temperature were maintained at 200 ℃. The fragmentation voltage was 70V. Mass spectra were obtained by scanning from 100 to 1000 amu in electrospray ionization or APCI in positive mode.

Method WUXI4: in addition to the general program WUXI: reverse phase HPLC was performed on an Agilent TC-C18 column (5 μm, 2.1 x50 mm) at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: water with 0.1% trifluoroacetic acid; mobile phase B: acetonitrile with 0.05% trifluoroacetic acid) were used to run the following gradient conditions: starting at 90% A/10% B, hold for 0.8 min, in 3.7 min to 20% A/80% B, hold for 3 min, and then return to the initial conditions in 2 min. The capillary voltage was 2.5 kV for the positive ionization mode and 3kV for the negative ionization mode, and the corona discharge was kept at 4 μ Α if APCI and source temperature were maintained at 200 ℃. The fragmentation voltage was 70V. Mass spectra were obtained by scanning from 100 to 1000 amu in electrospray ionization or APCI in positive mode.

General procedure Mercachem (for method MERCx)

HPLC measurements were performed using HPLC 1100-SL or 1200-SL (agilent) systems equipped with deaerators, autosamplers, Diode Array Detectors (DADs) and columns as specified in the methods below, containing quaternary pumps. The MS detector (Agilent MSD-SL) was configured with an electrospray ionization source. Data acquisition was performed using an Agilent Chemstation data system.

Method MERC20: in addition to the general procedure MERC: reverse phase HPLC was performed on a Waters X-BridgeC18 column (3.5 μm, 2.1X 50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: acetonitrile containing 10mM ammonia; mobile phase B: water containing 10mM ammonia) were used to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 2.5 min. Mass spectra were obtained in electrospray ionization in positive and negative modes, with scans from 220 to 800 amu.

Method MERC22: in addition to the general procedure MERC: reverse phase HPLC was performed on a Waters X-BridgeC18 column (3.5 μm, 2.1X 50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 95% methanol/5% 10mM ammonium bicarbonate in water; mobile phase B: 10mM ammonium bicarbonate in water) were employed to run the following gradient conditions: starting from 10% A to 98% A/2% B in 2.5 min, hold 3.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 220 to 800 amu.

Method MERC25: in addition to the general procedure MERC: reverse phase HPLC was performed on a Gemini C18 column (3 μm, 2.1 x50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 95% acetonitrile/5% 10mM ammonium bicarbonate in water; mobile phase B: 10mM ammonium bicarbonate in water) were employed to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 2.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 100 to 800 amu.

Method MERC26: in addition to the general procedure MERC: reverse phase HPLC on Waters X-BridgeC18 column (3.5 μm, 2.1 x50 mm) at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 0.1% formic acid in acetonitrile; mobile phase B: 0.1% formic acid in water) were employed to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 2.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 100 to 800 amu.

Method MERC27: in addition to the general procedure MERC: reverse phase HPLC was performed on a Waters X-BridgeC18 column (3.5 μm, 2.1X 50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 95% acetonitrile/5% 10mM ammonium bicarbonate in water; mobile phase B: 10mM ammonium bicarbonate in water) were employed to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 4.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 100 to 800 amu.

Method MERC28: in addition to the general procedure MERC: reverse phase HPLC was performed on a Waters X-BridgeC18 column (3.5 μm, 2.1X 50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 95% acetonitrile/5% 10mM ammonium bicarbonate in water; mobile phase B: 10mM ammonium bicarbonate in water) were employed to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 2.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 100 to 800 amu.

Method MERC30: in addition to the general procedure MERC: reverse phase HPLC was performed on a Gemini C18 column (3 μm, 2.1 x50 mm) maintained at 25 ℃ at a flow rate of 0.8 ml/min. Two mobile phases (mobile phase a: 95% acetonitrile/5% 10mM ammonium bicarbonate in water; mobile phase B: 10mM ammonium bicarbonate in water) were employed to run the following gradient conditions: starting from 2% A to 98% A/2% B in 3.5 min, hold 4.5 min. Mass spectra were acquired by electrospray ionization in positive and negative modes, scanning from 100 to 800 amu.

H of the final Compound¹NMR analysis:

compound 6

¹H NMR (500 MHz, DMSO-d₆) d 8.81 (d, J = 5.67 Hz, 2H)，8.77 (s, 1H)，8.31 (d, J = 5.67 Hz, 2H)，7.46 (t, J = 7.88 Hz, 1H)，7.07 (d, J = 7.88 Hz,1H)，6.98 - 7.04 (m, 2H)，3.81 (s, 3H)，2.79 - 2.89 (m, 3H)，1.96 - 2.15 (m, 6H)，1.66 (d, J = 11.98 Hz, 2H)，0.86 (s, 9H)

Compound 55

¹H NMR (500 MHz, DMSO-d₆)8.74 - 8.85 (m, 3H)，8.33 (d, J = 4.73 Hz,2H)，7.46 (t, J = 7.88 Hz, 1H)，6.97 - 7.11 (m, 3H)，3.82 (s, 3H)，3.16 (q, J =9.98 Hz, 2H)，2.99 (d, J = 11.03 Hz, 2H)，2.90 (t, J = 11.03 Hz, 1H)，2.27 (t, J= 11.66 Hz, 2H)，2.02 (q, J = 11.66 Hz, 2H)，1.72 (d, J = 11.66 Hz, 2H)

Compound 58

¹H NMR (400 MHz, CHLOROFORM-d)8.86 (d, J = 5.31 Hz, 2H)，8.67 (s, 1H)，8.44 (d, J = 5.31 Hz, 2H)，7.86 (d, J = 7.83 Hz, 1H)，7.67 - 7.76 (m, 2H)，7.64(d, J = 7.83 Hz, 1H)，3.02 - 3.17 (m, 4H)，2.80 (s, 1H)，2.37 - 2.47 (m, 2H)，2.22 - 2.37 (m, 2H)，1.75 (d, J = 13.39 Hz, 2H)

Compound 70

¹H NMR (400 MHz, CHLOROFORM-d) 8.85 (d, J = 6.06 Hz, 2H)，8.69 (s,1H)，8.41 - 8.46 (m, 2H)，7.48 (t, J = 8.08 Hz, 1H)，7.07 (dd, J = 2.02, 8.08Hz, 1H)，6.96 (d, J = 7.33 Hz, 1H)，6.90 (s, 1H)，3.93 (s, 3H)，3.03 - 3.14 (m,2H)，2.90 - 3.01 (m, 1H)，2.73 (t, J = 13.64 Hz, 2H)，2.18 - 2.32 (m, 4H)，1.74(s, 2H)，1.62 (br. s., 3H)

Compound 67

¹H NMR (400 MHz, CHLOROFORM-d)9.31 (d, J = 2.27 Hz, 1H)，8.60 (dd, J =2.27, 8.59 Hz, 1H)，8.43 (s, 1H)，7.69 (d, J = 7.58 Hz, 1H)，7.53 - 7.60 (m,2H)，7.45 - 7.52 (m, 1H)，6.80 (d, J = 8.59 Hz, 1H)，3.97 (s, 3H)，2.80 (d, J =6.82 Hz, 2H)，2.57 (br. s., 1H)，2.01 - 2.16 (m, 4H)，1.96 (s, 2H)，1.49 - 1.58(m, 2H)，0.81 (s, 9H)

Compound 57

¹H NMR (400 MHz, DMSO-d₆) 8.76 - 8.85 (m, 3H)，8.29 - 8.37 (m, 2H)，7.42- 7.52 (m, 1H)，6.97 - 7.12 (m, 3H)，3.83 (s, 3H)，3.02 (d, J = 11.12 Hz, 2H)，2.89 (t, J = 11.10 Hz, 1H)，2.44 (s, 2H)，2.14 - 2.27 (m, 2H)，1.94 - 2.12 (m,2H)，1.72 (d, J = 12.63 Hz, 2H)，1.29 (s, 6H)

Compound 66

¹H NMR (400 MHz, CHLOROFORM-d)8.62 (dd, J = 5.81, 8.84 Hz, 2H)，8.57(s, 1H)，7.82 (d, J = 7.58 Hz, 1H)，7.66 - 7.73 (m, 2H)，7.61 (s, 1H)，7.25 (t, J= 8.84 Hz, 2H)，2.94 (d, J = 7.58 Hz, 2H)，2.60 - 2.78 (m, 1H)，2.13 - 2.31 (m,4H)，2.09 (s, 2H)，1.64 (d, J = 8.34 Hz, 2H)，0.95 (s, 9H)

Compound 65

¹H NMR (400 MHz, CHLOROFORM-d) 8.73 (d, J = 8.34 Hz, 2H)，8.63 (s,1H)，7.80 - 7.90 (m, 3H)，7.66 - 7.74 (m, 2H)，7.60 - 7.65 (m, 1H)，2.94 (d, J =6.57 Hz, 2H)，2.65 - 2.79 (m, 1H)，2.14 - 2.30 (m, 4H)，2.09 (s, 2H)，1.65 (d, J= 4.80 Hz, 2H)，0.95 (s, 9H)

Compound 71

¹H NMR (400 MHz, CHLOROFORM-d)8.49 - 8.61 (m, 3H)，7.41 (t, J = 8.03Hz, 1H)，7.17 (t, J = 8.03 Hz, 2H)，7.00 (dd, J = 2.01, 8.03 Hz, 1H)，6.90 (d, J= 8.03 Hz, 1H)，6.81 - 6.86 (m, 1H)，3.87 (s, 3H)，2.84 - 3.11 (m, 5H)，2.16 -2.42 (m, 4H)，1.70 (d, J = 13.05 Hz, 2H)

Compound 39

¹H NMR (400 MHz, DMSO-d₆)8.73 - 8.90 (m, 3H)，8.32 (d, J = 6.06 Hz,2H)，7.53 - 7.68 (m, 3H)，7.40 - 7.50 (m, 1H)，2.84 (d, J = 11.12 Hz, 2H)，2.68 -2.79 (m, 1H)，1.92 - 2.17 (m, 6H)，1.66 (d, J = 12.13 Hz, 2H)，0.86 (s, 9H)

Compound 21

¹H NMR (400 MHz, DMSO-d₆)8.69 - 8.79 (m, 2H)，8.36 (s, 1H)，8.19 - 8.28(m, 2H)，7.34 - 7.48 (m, 1H)，7.04 - 7.15 (m, 2H)，6.97 (dd, J = 1.77, 8.34 Hz,1H)，3.81 (s, 3H)，3.36 - 3.40 (m, 3H)，2.46 (t, J = 4.55 Hz, 3H)，2.04 (s, 2H)，1.23 (br. s., 2H)，0.83 (s, 9H)

Compound 40

¹H NMR (400 MHz, DMSO-d₆)8.75 - 8.96 (m, 3H)，8.33 (d, J = 6.06 Hz,2H)，7.89 (br. s., 2H)，7.72 - 7.85 (m, 2H)，2.84 (d, J = 9.35 Hz, 2H)，2.63 -2.77 (m, 1H)，1.93 - 2.17 (m, 6H)，1.56 - 1.78 (m, 2H)，0.85 (s, 9H)

The following 6 compounds/examples were also prepared according to the procedures described herein:

。

biological examples

In vitro method for testing the antibacterial activity of compounds against various bacterial strains

Preparation of bacterial suspensions for susceptibility testing

The following bacteria were used: staphylococcus aureus ATCC 29213, methicillin-resistant staphylococcus aureus (MRSA) ATCC 700788, and Escherichia coli (Escherichia coli) ATCC 35218. The bacteria used in this study were grown overnight at 37 ℃ with shaking in a flask containing 100ml Mueller-Hinton broth (Difco cat. nr. 0757-17) in sterile deionized water. The stock solution was stored at-70 ℃ until use.

Bacteria were incubated on tryptic soy agar plates containing 5% sheep blood (Becton Dickinson cat. nr.254053) for 18-24 hours (first generation) under aerobic conditions at 35 ℃. For the second generation, 5-10 colonies were inoculated in fresh Mueller-Hinton broth and grown overnight at 35 ℃ under aerobic conditions until turbidity was reached (log phase reached). The bacterial suspension was then adjusted to 0.5 McFarland density and further diluted 1:100 in Mueller Hinton broth. It was used as an inoculum.

And (3) antibacterial susceptibility testing: IC90 determination

MIC analysis was performed by broth microdilution method on 96-well format (flat bottom microtiter plate) with a final volume of 0.1ml Mueller Hinton broth containing two-fold serial dilutions of the compound and inoculated with 5x105 CFU/ml bacteria (standard inoculum size according to CLSI guidelines). Inhibitors typically vary in the range of 63-0.49 μ M. The final DMSO concentration in the assay was 1.25% (maximum tolerable DMSO concentration = 6%). In an assay to test the effect of human serum on the activity of compounds against staphylococcus aureus, human serum was added at a final concentration of 10%. The plates were incubated at 35 ℃ for 16-20 hours. At the end of the incubation, the bacterial growth was quantified by fluorometric analysis. For this purpose, resazurin was added to all wells and the plates were incubated again. The incubation time depends on the type of bacteria. The color changed from blue to pink, indicating the growth of bacteria. In a computer-controlled fluorometer (Fluoroskan Ascent FL, Labsystems), the fluorescence is read off at an excitation wavelength of 540 nm and an emission wavelength of 590 nm. The% growth inhibition achieved by the compounds was calculated according to standard methods. IC90 (expressed in μ g/ml) was defined as the 90% inhibitory concentration for bacterial growth. A panel of reference compounds was assayed simultaneously for QC approval.

Cytotoxicity assays

Cytotoxicity of compounds was assessed using MTT assay. Human HelaM cells grown in 96-well plates were exposed to serial dilutions of test compounds (final volume 0.2 ml) and at 37 ℃ and 5% CO₂Incubate for 72 hours. Inhibitors typically vary in the range of 25 to 0.8 μ M. The final DMSO concentration in the assay was 0.5%. Adding MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazole)Bromide, a tetrazole) and reduced to purple formazan only in living cells(formalzan). First of allDissolution of the crystals was obtained by adding 100 μ l of 2-propanol. Cell viability reduced formazan producing purple color as determined by 540 nm and 690nmDetermining the absorbance of (a). The absorbance measured at 690nm was automatically subtracted from the absorbance at 540 nm to eliminate the effect of non-specific absorption. The percent cytotoxicity achieved by the compounds was calculated according to standard methods. Cytotoxicity was reported as CC50, the concentration that caused a 50% reduction in cell viability.

Protocol for MIC determination of Compounds on ECO/PAE/STA on Microplate

Add 4-5 colonies of the overnight growth plate to 5 ml of Mueller Hinton medium

Culturing at 37 ℃ for 3-6 hours in a shaking incubator (300 rpm)

Measuring OD at 600 nm (OD600 = 1- - >109 CFU/ml)

Dilute bacteria until 105 CFU/ml in culture medium

In a microplate, 2-fold dilutions (final concentration from 64 to 0.125 mug/ml) were prepared in 100 mul Mueller Hinton medium

100 μ l of bacterial dilution was added to each well

Culturing at 37 ℃ for 18-20 hours

Visual inspection of growth relative to control

MIC is the lowest concentration without growth (90% growth inhibition)

Biological results

The example compounds/compounds of the invention were tested in the above described antibacterial susceptibility and/or cytotoxicity assays. It was found that in each analysis, example compounds/compounds of the invention showed an IC90 value of less than 50 μ g/mL (e.g. less than 15 μ g/mL), a CC50 value of less than 50 μ g/mL (e.g. less than 15 μ g/mL) and/or a MIC90 of less than 10 μ g/mL (e.g. less than 1 μ g/mL). In various assays, certain compounds exhibited IC90 values of less than 10 μ g/mL (e.g., less than 1 μ g/mL), or CC50 values of less than 10 μ g/mL (e.g., less than 5 μ g/mL) and/or MIC90 values of less than 0.5 μ g/mL.

Certain compounds are available from commercially available sources, such as CHEMBRIDGE.

TABLE 1 Compounds of formula (I)

TABLE 2 Compounds of formula (I)

Claims (8)

1. A compound of the formula I,

wherein:

y represents:

；

N^v、N^w、N^x、N^yand N^z0 or 1 out represents-N = and the others represent-c (h) =;

n represents 0 or 1;

X¹represents-N-;

X²represents-C (H) -;

Q¹represents a direct bond;

R^xis represented by C_1-6Alkyl optionally substituted with one or more groups selected from = O and A¹Substituted with the substituent(s);

R^y、R^y1and R^y2All represent hydrogen, or R^yRepresents halogen, -OCH₃or-CN and R^y1And R^y2Represents hydrogen;

A⁴represents halogen, -CN or-OC_1-3An alkyl group;

A¹represents halogen, -CN, C_1-6Alkyl OR-OR¹(ii) a And

R¹represents hydrogen, and is represented by the formula,

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a⁴Represents fluorine.

3. The compound of claim 1, wherein the compound is:

or a pharmaceutically acceptable salt thereof.

4. An antibacterial pharmaceutical combination comprising (a) a compound as defined in any one of claims 1 to 3, and (b) one or more other antibacterial agents.

5. A combined preparation for simultaneous, separate or sequential use in the treatment of a bacterial infection, which comprises (a) a compound as defined in any one of claims 1 to 3, and (b) one or more other antibacterial agents.

6. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound as defined in any one of claims 1 to 3.

7. Use of a compound as defined in any one of claims 1 to 3 in the manufacture of a medicament for the treatment of a bacterial infection.

8. The use of claim 7 wherein the bacterial infection is caused by Staphylococcus aureus.