WO1997004770A1 - 2-(fluorenonyl)-carbapenems, compositions and methods of use - Google Patents

Abstract

Carbapenems are disclosed in which the 2-position sidechain contains a fluorenone moiety that is substituted by a bis cationic group. The compounds are of general formula (I). The compounds are effective for treating methicillin resistant staphylococcus aureus (MRSA) and methicillin resistant coagulase negative staphylococcus (MRCNS). Pharmaceutical compositions and methods of treatment are also included.

Description

TITLE OF THE INVENTION

2-(FLUORENONYL)-CARBAPENEMS, COMPOSITIONS AND METHODS OF USE The present invention relates to antibacterial agents of the carbapenem class, in which the 2-position sidechain contains a fluorenone moiety which is substituted by a bis cationic group.

The bis cationic group is substituted with a variety of substituents.

U. S. Patent Number 5,034,384 issued on July 23, 1991 discloses 2- and 3-fluoren-9-onyl-2-carbapenems as having anti-MRSA/MRCNS activity.

The carbapenems of the present invention are characterized by an antibacterial spectrum which is largely focused on gram positive microorganisms, especially methicillin resistant Staphylococcus aureus (MRSA), methicillin resistant Staphylococcus epidermidis (MRSE), and methicillin resistant coagulase negative Staphylococci (MRCNS).

There is an increasing need for agents which are effective against these pathogens (MRSA/MRCNS) and which are considered safe, i.e., relatively free from undesirable side effects. The current agent of choice for the treatment of infections caused by MRSA, vancomycin, a glycopeptide antibacterial, is known to cause adverse reactions in many patients. The antibacterial compounds of the present invention thus comprise an important contribution to therapy of these difficult to control pathogens.

SUMMARY OF THE INVENTION

A compound represented by the formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R represents H or methyl;

A' and A" independently represent -(CH₂)_n-, wherein n is an integer of from 1 to 4;

X- represents a pharmaceutically acceptable counterion;

M represents a negative charge, H or an ester forming group;

Q' is selected from the group consisting of: a direct bond, -O-, -S-, -C(O)-, -CO(O)-, -OC(O)-, -C(O)N(R^z')-, -N(R^z')C(O)-, -SO₂-, -SO-, -SO₂N(R^z')-, -N(R^z')SO₂-, -OC(O)N(R^z')- and

-N(R^z')C(O)O-;

(a) when Q' represents -S-, -C(O)-, -OC(O)-, -C(O)N(R^z')-, -N(R^z')C(O)-, -N(R^z')SO₂- or -N(R^z')C(O)O-, R^e' represents a member selected from the group consisting of:

C₁ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₆ to C₁₄ aryl unsubstituted or substituted with 1 -3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5-10 atoms, 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups;

(b) when Q' represents -C(O)O-, R^e' represents a member selected from the group consisting of:

C₂ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10}) cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₈ to C₁₄ aryl, unsubstituted or substituted with 1 -3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1 -3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5 - 10 atoms, 1-3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups; (c) when 0' represents -O-, -SO-, -SO₂-, -SO₂N(R^z')- or -OC(O)N(R^z')-, R^e' is selected from the group consisting of:

C₂ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₆ to C₁₄ aryl, unsubstituted or substituted with 1-3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1 -3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5 - 10 atoms, 1-3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups; and (d) when 0' represents a direct bond, R^e' represents a member selected from the group consisting of:

C₄ to C₁₀ alkyl (including C_{4- 10} branched and C_{4- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups; C₆ to C₁₄ aryl, unsubstituted or substituted with 1-3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5-10 atoms, 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups;

R^z' is selected from the group consisting of: H, OCH₃, C₁ to C₄ alkyl and C₁ to C₄ alkyl substituted with 1 -3 R^q' groups, and each R^q' is independently selected from the group

consisting of: "OH, -OCH₃, -CN, -C(O)NH₂, -C(O)NHCH₃,

-C(O)N(CH₃)2, -OC(O)NH₂, -CHO, -SO₂NH₂, -SOCH₃, -SO₂CH₃, -F, -CF₃, -C(O)CH₃, -COOCH₃ and -COOM^a (where M^a is H, a negative charge or an alkali metal). Included in the invention is a pharmaceutical composition, which is comprised of a compound of formula I in combination with a pharmaceutically acceptable carrier.

Also included in the invention is a method of treating a bacterial infection comprising administering to a mammalian patient in need of such treatment an anti-bacterial effective amount of a compound of formula I.

DETAILED DESCRIPTION OF THE INVENTION

The invention is described herein in detail using the terms defined below unless otherwise specified.

The term "alkyl" and the alkyl portion of alkaryl and aralkyl refer to a monovalent alkane (hydrocarbon) derived radical containing from 1 to 10 carbon atoms unless otherwise specified. It may be straight, branched or cyclic. Preferred alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, t-butyl, cyclopentyl and cyclohexyl. When substituted, alkyl groups may be substituted with up to three substituents R^q', as defined, at any available point of attachment. When the alkyl group is said to be substituted with an alkyl group, this is used interchangeably with "branched alkyl

group".

Cycloalkyl is a specie of alkyl containing from 3

to 15 carbon atoms, without alternating or resonating double bonds between carbon atoms. It may contain from 1 to 4 rings which are fused.

Aryl and the aryl portion of aralkyl and alkaryl refer to aromatic rings e.g., phenyl, substituted phenyl and the like, as well as rings which are fused, e.g., naphthyl, phenanthrenyl and the like.

Aryl thus contains at least one ring having at least 6 atoms, with up to five such rings being present, containing up to 22 atoms therein, with alternating (resonating) double bonds between adjacent atoms. The preferred aryl groups are phenyl, naphthyl and phenanthrenyl.

Aralkyl is a specie of alkyl substituted with an aryl group. Alkaryl is a specie of aryl substituted with an alkyl group.

The aryl and alkyl portions of aralkyl and alkaryl may be substituted as described throughout the specification.

The term "heteroaryl" refers to a monocyclic aromatic hydrocarbon group having 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing at least one heteroatom, O, S or N, in which a carbon or nitrogen atom is the point of attachment, and in which one additional carbon atom is optionally replaced by a heteroatom selected from O or S, and in which from 1 to 3

additional carbon atoms are optionally replaced by nitrogen heteroatoms, said heteroaryl group being optionally substituted with up to two R^q' groups.

Heteroaryl thus includes aromatic and partially aromatic groups which contain one or more heteroatoms. Additional nitrogen atoms may be present together with the first nitrogen and oxygen or sulfur, giving, e.g., thiadiazole. The preferred heteroaryls are those where only nitrogen heteroatoms are present when there is more than one. Examples include pyrazole, tetrazole, imidazole, pyridine, pyrimidine and pyrazine and triazine.

The term "quaternary nitrogen" refers to the tetravalent positively charged nitrogen atoms present in

diazoniabicyclo[2.2.2]octanyl moiety.

The term "heteroatom" means N, S, or O, selected on an independent basis.

Alkylene (alkylidene or alkanediyl) and arylene refer to the groups noted above with divalent points of attachment. For example, phenylene is an arylene group, attached at any of the 1 , 2- 1 , 3- or 1 , 4- positions. Examples of alkylene include -CH₂-,

-CH₂CH₂-, -CH₂CH₂CH₂-,

Dialkylene and trialkylene refer to two and three of the above alkylene groups, respectively. With respect to the preferred compounds described herein, the preferred alkylene groups are methylene, ethylene, diethylene and triethylene.

Similarly, alkanetriyl and tertiary alkylidene refer to an alkane-derived group with three points of attachment. Alkanetriyl groups contain from five to fifteen carbon atoms, which may be straight, branched, cyclic or multicyclic. Aralkyl is a specie of substituted alkyl, containing up to three aryl groups substituted on a straight, branched or cycloalkyl group. The most preferred aralkyl group is benzyl.

Halogen, or "halo" refers to bromine, chlorine, fluorine and iodine.

A subset of compounds of the present invention is represented by formula la:

wherein all variables are as previously defined.

A more preferred subset of compounds of the present invention is represented by formula lb:

wherein all variables are as previously defined.

Another more preferred subset of compounds of the present invention is represented by formula Ic:

wherein all variables are as previously defined.

The preferred values of A' and A" are selected from -(CH₂)_n- wherein n represents 1 or 2, and most preferably, n equals 1.

The preferred value of R is methyl.

The preferred values of Q' are a direct bond, -C(O)- and -C(O)N(R^z')- .

The preferred value of R^e' are C₁ to C₃ alkyl, substituted with 1 R^q' group, and phenyl substituted with 1 R^q' group. When R^z' is present, the preferred values of R^z' are selected from the group consisting of: H, C₁-4 alkyl and C₁-4 alkyl substituted with 1 -3 R^q' groups.

The preferred value of R^q' is selected from the group consisting of: -OH, -CN, -C(O)NH₂, -CHO, -SO₂NH₂, -SOCH₃,

-SO₂CH₃, -F, -CF₃ and -C(O)CH₃. Most preferably, the value of R^q' is selected from the group consisting of: "OH, -CN and -C(O)NH₂.

Preferred species falling within the scope of the present invention are as follows:

As will be recognized in the tables, R^a which appears in the tables represents the group:

The starting marterials for the syntheses described herein can be made according to the scientific and patent literature. For example, the steps for preparing the 2-oxocarbapenem A2 where R=H are well known in the art and are explained in ample detail by D. G. Melillo et al., Tetrahedron Letters, 1980. 21 , 2783; T. Salzmann et al., J. Am. Chem. Soc., 1980, 102, 6161 ; and L. M. Fuentes, I. Shinkai and T. N. Salzmann, J. Am. Chem. Soc., 1986, 108, 4675. The syntheses are also disclosed in U. S. Patent No. 4,269,772, U. S. Patent No.

4,350,631 , U. S. Patent No. 4,383,946 and U. S. Patent No. 4,414,155 all assigned to Merck and Company, Inc. Details pertaining to the formation of the 1-β-methyl intermediates A1 and A2 (R=CH₃) and precursors thereto can be found in Shih, D. H., et al. Heterocycles 21 : 29 (1984) and in Fuentes, L. M. et al. J. Am. Chem. Soc., 108, 4675 (1986). The syntheses are also disclosed in U.S. Pat. No. 4,269,772, U.S. Pat. No. 4,350,631 , U.S. Pat. No. 4,383,946 and U.S. Pat. No. 4,414,155 and U. S. Pat. No. 4,994,568, all assigned to Merck and Co., Inc.

FLOW SHEET A

Briefly, as shown in Flow Sheet A, the diazo-β-ketoester A1 is cyclized to form the 2-oxocarbapenam A2 , which is thereafter activated at the 2-position by conversion to the enol trifluoromethanesulfonate derivative and protected in the hydroxyethyl side-chain to form A3. Intermediate A3, is then coupled with an appropriate fluorenone synthon A4 to produce the carbapenem intermediate A5.

The diazo-β-ketoester A1 is cyclized by heating at from about 15°C to about 50°C from about one to four hours in a suitable inert solvent such as dichloromethane tetrahydrofuran or chloroform in the presence of a suitable transition metal catalyst such as rhodium(II) octanoate [Rh₂(Oct)₄] or rhodium(II) acetate [Rh₂(OAc)₄] to provide the 2-oxocarbapenam A2. The intermediate A2 can be reacted in situ with a suitable organic nitrogen base such as triethylamine, diisopropylethylamine, diisopropylamine and the like, followed by a suitable trifluoromethanesulfonylating agent such as trifluoromethanesulfonic anhydride, trifluoromethanesulfonyl chloride and the like, at reduced temperature such as from about -78°C to -20°C for about five to forty-five minutes. The hydroxyethyl side-chain of the resulting enol trifluoromethansulfonate intermediate can then be protected by introduction of an appropriate protecting group. For example, a suitable organic nitrogen base such as triethylamine, diisopropylethylamine, diisopropylamine or the like is then added to the reaction solution followed by a silylating agent such as triethylsilyl or trimethylsilyl trifluoromethanesulfonate to provide, after a reaction period of about five minutes to about two hours, the trifluoromethanesulfonate intermediate A3. The activated 2-(trifluoromethanesulfonyloxy)carbapenem intermediate A3 is thereafter coupled with an appropriately substituted fluoren-9-one A4 as described further below.

The synthesis of the fluoren-9-one A4 can be varied depending upon the value of Met. When Met represents a trialkyltin moiety and n equals 1 , the fluoren-9-one is synthesized according to the process described in U.S. Pat. Appl. 485,096 filed February 26, 1990, and in U. S. Pat. No. 5,034,384 issued on July 23, 1991. Briefly, 6-bromo-2-hydroxymethylfluoren-9-one is reacted with hexamethylditin, tetrakis(triphenyl-phosphine) palladium(0) and triphenylphosphine in toluene at about 110°C to produce 6-trimethylstannyl-2-hydroxymethylfluoren-9-one. An aprotic polar coordinating solvent, such as N,N- dimethylformamide, 1-methyl-2-pyrrolidinone and the like, can also be added. Compounds wherein n equals 2 - 4 can be made with reference to the literature without resort to undue experimentation.

When Met represents a boronic acid moiety and n equals 1, the fluoren-9-one is synthesized according to U. S. Pat. Application No. 978,598 filed on November 19, 1992 and copending herewith. Briefly, 3-bromo-9,9-dimethoxy-7-methoxymethylfluorene is reacted with n-butyllithium and the metalated fluorene is boronated with triisopropyl borate [B(Oi-Pr)3]. After hydrolysis and removal of protecting groups, 2-hydroxymethylfluoren-9-one-6-boronic acid is obtained.

The conditions of the coupling reaction between the activated carbapenem A3 and the fluorenone A4 vary depending upon the value of Met. When Met is a trialkyltin moiety, a solution of 2-(trifluoromethanesulfonyloxy)carbapenem A3, in a suitable solvent such as tetrahydrofuran, 1-methyl-2-pyrrolidinone, N,N-dimethylformamide or dichloromethane is combined with a palladium compound, e.g., tris(dibenzylideneacetone)dipalladium-chloroform, bis(dibenzylideneacetone)palladium, palladium acetate, bis(acetonitrile)palladium(Il) chloride and the like, optionally a suitably substituted phenylphosphine, such as tris(4-methoxyphenyl)phosphine, tris(2,4,6-trimethoxyphenyl) phosphine and the like, and the trialkylstannyl-fluorenone A4 . A metal halide, such as lithium chloride, zinc chloride and the like or an ammonium halide such as tetraethylammonium chloride, diisopropylammonium hydrochloride and the like, is added and the reaction solution is maintained at a suitable temperature, such as from about 0°C to 50°C, and allowed to stir for a suitable amount of time such as from a few minutes to about 48 hours. The carbapenem A5 is thereafter obtained by conventional isolation/purification methodology known in the art.

When Met is a boronic acid moiety, the carbapenem A3 and boronic acid A4 are combined in a coupling solvent with a coupling base and a transition metal catalyst as described in U. S. Application No. 978,598.

Coupling bases for purposes of this reaction include metal hydroxides, metal C_1-4 alkoxides and metal carbonates. Examples of metal hydroxides include barium, potassium, sodium, lithium and thallium hydroxide. Examples of metal alkoxides include sodium, potassium and lithium t-butoxide. Examples of metal carbonates include potassium and sodium carbonate.

Coupling solvents for purposes of this reaction include di-C_{1 -3} alkyl formamides, di-C_{1 -3} alkyl sulfoxides , N-alkylpyrrolidinones, halocarbons, ethers, aromatic and aliphatic solvents. An example of a di-C_{1 -3} alkyl formamide is N,N-dimethylformamide. An example of a di-C_{1 -3} alkyl sulfoxide is dimethylsulfoxide. Examples of N-alkylpyrrolidones include N-methylpyrrolidone and N-ethylpyrrolidone. An example of a halocarbon is dichloromethane. Examples of ethereal solvents include diethyl ether, di-n-butyl ether, tetrahydropyran and tetrahydrofuran. Examples of aromatic solvents include benzene, toluene and xylene. Examples of aliphatic solvents include n-hexane and cyclohexane.

Transition metal catalysts for purposes of this reaction include palladium and nickel catalysts. Examples of palladium

catalysts include Pd(0) and Pd(II) catalysts. The Pd(0) catalysts include tris(dibenzylideneacetone)dipalladium-chloroform,

tris(dibenzylideneacetone)dipalladium and bis(dibenzylideneacetone)-palladium. Examples of Pd(II) catalysts include Pd(OAc)₂ and PdCl₂.

The boronic acid derivative A4 is added as a solution in a suitable polar solvent such as 1 -methyl-2-pyrrolidinone, N,N-dimethylformamide or tetrahydrofuran to a solution of 2-(trifluoromethanesulfonyloxy)carbapenem A3 in a suitable solvent such as 1 -methyl-2-pyrrolidinone, N,N-dimethylformamide, tetrahydrofuran or dichloromethane followed by the addition of a palladium compound such as tris(dibenzylideneacetone)dipalladium-chloroform,

bis(dibenzylideneacetone)palladium, palladium acetate, bis(acetonitrile) palladium(II) chloride and the like, optionally a phosphine such as triphenylphosphine, and an inorganic base such as aqueous potassium hydroxide, aqueous potassium carbonate, aqueous cesium hydroxide, or solid potassium carbonate. The reaction solution is maintained at a suitable temperature, such as from about 0 °C to 60 ºC, and allowed to stir for a suitable amount of time such as from a few minutes to 48 hours. The carbapenem A5 is then obtained by conventional

isolation/purification methodology known in the art.

While the intermediates A2 and A3 may be isolated by conventional means, it is preferable in the case of R=H to carry-out the sequence A2 to A5 in situ and in the case of R=CH₃ to carry-out the entire sequence of Flow Sheet A in situ . Conversion of the carbapenem intermediate A5 into the final compound I may be accomplished as shown in Flow Sheet B. Briefly, the hydroxyl group of A5 is converted into a suitable leaving group, Z, which is thereafter displaced with a 1 -substituted-4-aza-1 -azoniabicyclo(2.2.2)octane salt B2 to provide B3. The protecting groups are removed from B3 in conventional fashion and then the desired counterion X- is introduced to provide compound I.

The following are examples of suitable leaving groups Z: alkyl and substituted alkylsulfonates, aryl and substituted arylsulfonates and halides. The common sulfonate leaving groups are: methanesulfonyloxy, trifluoromethanesulfonyloxy, fluorosulfonyloxy, p-toluenesulfonyloxy, 2,4,6-triisopropylbenzenesulfonyloxy,

p-bromobenzenesulfonyloxy and p-nitrobenzenesulfonyloxy.

The preferred halogen leaving groups are bromide and iodide.

Referring to Flow Sheet B, the hydroxyl group of A5 may be converted into a suitable alkyl- or arylsulfonate leaving group by treating with an appropriate agent such as an alkyl- or arylsulfonyl chloride or an alkyl- or arylsulfonic anhydride in the presence of a hindered organic base such as triethylamine. A suitable solvent such as dichloromethane is employed and the reaction is carried out at reduced temperature, such as from about -70°C to 0°C.

FLOW SHEET B

The preferred halogen leaving groups may be introduced by displacing an alkyl- or arylsulfonate leaving group with an appropriate metal halide. Thus, compound B1, where Z is an alkyl- or arylsulfonate group, is reacted with a suitable metal halide such as sodium iodide or potassium bromide in a suitable solvent such as acetone, acetonitrile, tetrahydrofuran, 1-methyl-2-pyrrolidinone and the like, at from about 0 °C to 50 °C. Alternatively, the hydroxyl group of A5 may be directly converted into an iodide group by reaction with an appropriate reagent, eg. by treatment of A5 with methyl triphenoxyphosphonium iodide in a suitable solvent, such as N,N-dimethylformamide, at reduced or ambient temperatures.

Introduction of the cationic substituent is accomplished by reacting B1 with a 1-substituted-4-aza-1-azoniabicyclo(2.2.2)octane salt B2 in a suitable solvent, such as acetonitrile, tetrahydrofuran, 1-methyl-2-pyrrolidinone and the like, at about 0 °C to 50 °C to provide B3.

When the leaving group, Z, is iodide or bromide, this displacement reaction may also be facilitated by the addition of silver

trifluoromethanesulfonate to the reaction mixture.

When the reactive trifluoromethanesulfonate group is employed as the leaving group Z in B1 and n equals 1 , the activation and displacement steps must be carried-out in situ, since in this case B1 cannot be isolated by conventional techniques due to its instability.

Thus, treatment of A5 with a slight excess of trifluoromethanesulfonic anhydride in the presence of a hindered, non-nucleophilic base such as 2,6-lutidine, 2,4,6-collidine, or 2,6-di-tert-butyl-4-methyl-pyridine in a suitable solvent, such as dichloromethane or acetonitrile, at from about -78 °C to -20 °C provides for the generation of the trifluoromethanesulfonate activating group. Introduction of the bis-quaternary

ammonium group is then accomplished by reacting the above

trifluoromethanesulfonate intermediate in situ with B2 at reduced temperature.

The amine B2 is prepared by reaction of 1 ,4-diazabicyclo[2.2.2]octane with one equivalent of an appropriate alkylating agent, Z'-A"-Q'-Re', where Z' is a leaving group, in an appropriate solvent such as acetonitrile, tetrahydrofuran or 1-methyl-2-pyrrolidinone. The following are examples of suitable leaving groups Z': alkyl and substituted alkylsulfonates, aryl and substituted arylsulfonates and halides. The common sulfonate leaving groups are: methanesulfonyloxy, trifluoromethanesulfonyloxy, fluorosulfonyloxy, p-toluenesulfonyloxy, 2,4,6-triisopropylbenzenesulfonyloxy, p-bromo benzenesulfonyloxy and p-nitrobenzenesulfonyloxy. The preferred halogen leaving groups are bromide and iodide. When Z' is halide, it is sometimes desirable to replace the halide counterion in B2 with a sulfonate counterion to improve solubility in organic solvents. For example a bromide counterion in B2 may be replaced with a trifluoromethanesulfonate counterion by reaction with a trifluoromethanesulfonate salt such as silver or sodium trifluoromethanesulfonate in an appropriate solvent such as acetonitrile, methanol or water. The alkylating agents Z'-A"-Q'-Re' are commercially available or may be prepared from readily available starting materials by methods well known to those skilled in the art.

In the preparation methods described herein, the carboxyl group at the 3-position and the hydroxyl group at the 8-position of the carbapenem typically remain blocked until the final product is prepared. These blocking groups are readily removable, i.e., they can be

removed, if desired, by procedures which will not cause cleavage or other disruption of the remaining portions of the molecule. Such procedures include chemical and enzymatic hydrolysis, treatment with chemical reducing or oxidizing agents under mild conditions, treatment with fluoride ion, treatment with a transition metal catalyst and a nucleophile, and catalytic hydrogenation.

Examples of suitable hydroxyl protecting groups are:

t-butylmethoxyphenylsilyl, t-butoxydiphenylsilyl, trimethylsilyl, triethylsilyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, benzyloxycarbonyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl and allyloxycarbonyl. Preferred hydroxyl protecting groups are trimethylsilyl and triethylsilyl.

Examples of suitable carboxyl protecting groups are:

benzhydryl, o-nitrobenzyl, p-nitrobenzyl, 2-naphthylmethyl, allyl, 2-chloroallyl, benzyl, 2,2,2-trichloroethyl, trimethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, 2-(trimethylsilyl)ethyl, phenacyl, p-methoxybenzyl, acetonyl, p-methoxyphenyl, 4-pyridylmethyl and t-butyl. A preferred carboxyl protecting group is p-nitrobenzyl. Many other suitable hydroxyl and carboxyl protecting groups are known in the art. See, e.g., Greene, T. W., et al. Protective Groups in Organic Synthesis. John Wiley & Sons, Inc., 1991.

Removal of the protecting groups of B3 where P is triethylsilyl and M is p-nitrobenzyl is accomplished by exposing B3 to aqueous acidic conditions, such as dilute hydrochloric acid, in an organic solvent such as tetrahydrofuran at from 0 °C to 30 °C for a few minutes to several hours. The resulting desilylated carbapenem may be isolated by conventional techniques, but is more conveniently taken directly into the final deprotection process. Thus, the reaction mixture is neutralized by addition of an inorganic base such as sodium

bicarbonate or sodium hydroxide and optionally a pH 6.5 to pH 7.0 aqueous buffer such as 4-morpholinepropanesulfonic acid/NaOH

(MOPS) or NaH₂PO₄/Na₂HPO₄. The reaction mixture is then hydrogenated at or slightly above atmospheric pressure over a heterogeneous catalyst such as rhodium on carbon, rhodium on alumina, palladium on carbon or the like at from 0 °C to 30 °C for from 30 minutes to 6 hours to remove the p-nitrobenzyl ester protecting group.

After the protecting groups are removed from B3, the desired counterion X- may be introduced by standard techniques, e.g. by employing an anion exchange resin or by utilizing the principle of mass action, i.e. exposure of compound I to a large excess of the desired anion. For example, introduction of the chloride counterion may be accomplished by dissolving compound I in a solution containing a large excess of sodium chloride. The final compound I where X- = Cl- is then isolated by conventional techniques.

Flow Sheet C illustrates an alternative method for introduction of the 1 ,4-diazoniabicyclo[2.2.2]octanyl group. Briefly, the hydroxyl group of A5 is converted into a suitable leaving group, Z, as described in Flow Sheet B giving B1. Reaction of B1 with with 1 ,4-diazabicyclo[2.2.2]octane provides C1 which is alkylated with an appropriate reagent Z'-A"-Q'-R^e' to give C2. It is recognized that depending on the identity of the counterions Z- and Z'^_, C2 may be the same as B3. Removal of the protecting groups from C2 and introduction of the desired counterion X- as described above for Flow Sheet B provides the final compound I.

FLOW SHEET C

Referring to Flow Sheet C, introduction of the cationic substituent is accomplished by reacting B1 with 1 ,4-diazabicyclo[2.2.2] octane in a suitable solvent, such as acetonitrile, tetrahydrofuran, 1 - methyl-2-pyrrolidinone dichloromethane and the like, at about 0 °C to 50 °C to provide C1. When the leaving group, Z, is iodide or bromide, this displacement reaction may also be facilitated by the addition of silver trifluoromethanesulfonate to the reaction mixture.

When the leaving group, Z, in B1 is trifluoromethanesulfonate and n equals 1 , the activation and displacement steps must be carried-out in situ, since in this case B1 cannot be isolated by conventional techniques due to its instability. Thus, treatment of A5 with a slight excess of trifluoromethanesulfonic anhydride in the presence of a hindered, non-nucleophilic base such as 2,6-lutidine, 2,4,6-collidine, or 2,6-di-tert-butyl-4-methyl-pyridine in a suitable solvent, such as dichloromethane or acetonitrile, at from about -78°C to -20 °C provides for the generation of the trifluoromethanesulfonate activating group. Introduction of the 4-aza-1 -azoniabicyclo(2.2.2) octanyl group is then accomplished by reacting the trifluoromethanesulfonate intermediate in situ with 1 ,4-diazabicyclo[2.2.2]octane at reduced temperature such as from about -78 °C to 0 °C to provide intermediate C1.

It is also possible to use the reacting amine as the base for the formation of the trifluoromethanesulfonate activating group. In this case, treatment of A5 with trifluoromethanesulfonic anhydride in the presence of at least two equivalents of 1 ,4-diazabicyclo[2.2.2]octane at reduced temperature such as from -78°C to 0°C provides intermediate C1.

Compound C1 is reacted with one equivalent of an appropriate alkylating agent, Z'-A"-Q'-R^e', where Z' is a leaving group, in an appropriate solvent such as acetonitrile, tetrahydrofuran or 1-methyl-2-pyrrolidinone and the like at from about 0 °C to 50 °C, to provide C2. The following are examples of suitable leaving groups Z': alkyl and substituted alkylsulfonates, aryl and substituted arylsulfonates and halides. The common sulfonate leaving groups are: methanesulfonyloxy, trifluoromethanesulfonyloxy, fluorosulfonyloxy, p-toluenesulfonyloxy, 2,4,6-triisopropylbenzenesulfonyloxy, p-bromobenzenesulfonyloxy and p-nitrobenzenesulfonyloxy. The preferred halogen leaving groups are bromide and iodide. When the leaving group, Z', is iodide or bromide, the alkylation reaction may be facilitated by the addition of silver trifluoromethanesulfonate to the reaction mixture. The alkylating agents Z'-A"-Q'-R^e' are commercially available or may be prepared from readily available starting materials by methods well known to those skilled in the art.

Removal of the protecting groups from C2 and introduction of the desired counterion X- as described above for Flow Sheet B provides the final compound I.

The compounds shown in the Flow Sheets and the final compounds are electronically balanced. Since a bis-quaternary

ammonium group is present in B3, and in the final product, for example, when M is H or an ester forming group, two negatively charged counterions are also present to provide overall electronic balance. One of these counterions is Z'- which derives from B2. The other counterion, Z-, varies depending on which activating group Z is employed. When a halogen activating group Z is used in B1 and the displacement with B2 is carried out in the presence of silver

trifluoromethanesulfonate, then Z- is normally trifluoromethanesulfonate. Likewise, the counterion Z- in C1 and C2 varies depending on which activating group Z is employed in BL The counterion Z'- in C2 varies according to the alkylating agent Z'-A"-Q'-R^e' that is used.

In the final compound I, when M represents a negative charge on the carboxylate, the charge of the bis-quaternary ammonium group is balanced by a negatively charged counterion, X-, in conjunction with the negatively charged carboxylate, CO₂-, which is contained in the molecule. Counterion X- is a pharmaceutically acceptable anionic species and may differ from Z- and Z'-. The desired counterion X- may be introduced by standard techniques as described above. It is understood that when the counterion X- is an anionic species possessing more than one negative charge, then an appropriate amount of X- is present to result in overall electronic balance in the final compound I. For example, when X- is a dianionic species, then one-half of a molar equivalent of X- is present relative to the

carbapenem moiety. Suitable negatively charged counterions, X-, are listed below under the description of pharmaceutically acceptable salts.

The Flow Sheets depicted above illustrate a particular isomeric attachment of the carbapenem and the cationic substituent to the fluorenone moiety. Other isomeric arrangements about the fluorenone ring system may be obtained by starting with the appropriate isomer of A4 in Flow Sheet A to produce the desired isomeric A5.

The carbapenem compounds of the present invention are useful in various pharmaceutically acceptable salt forms for the treatment of bacterial infections in animal and human subjects. The term "pharmaceutically acceptable salt" refers to those salt forms which would be apparent to the pharmaceutical chemist, i.e., those which are substantially non-toxic and which provide the desired pharmacokinetic properties, palatability, absorption, distribution, metabolism or excretion. Other factors, more practical in nature, which are also important in the selection, are cost of the raw materials, ease of crystallization, yield, stability, hygroscopicity and flowability of the resulting bulk drug. Conveniently, pharmaceutical compositions may be prepared from the active ingredients in combination with pharmaceutically acceptable carriers. Thus, the present invention includes pharmaceutical compositions and methods of treating bacterial infections utilizing the carbapenem compound of formula I.

The pharmaceutically acceptable salt forms of the carbapenem compounds of formula I mentioned above refer to the various possibilities for the charge balancing counterion X-. Anions derived from inorganic or organic acids are suitable. Representative examples of such counterions are the following: acetate, adipate, aminosalicylate, anhydromethylenecitrate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, bromide, citrate, camphorate, camphorsulfonate, chloride, digluconate, edetate, edisylate, estolate, ethanesulfonate, fumarate, glucoheptanoate, gluconate, glutamate, glycerophosphate, glycolate, hydroxynaphthoate, 2-hydroxyethanesulfonate, iodide, lactate, lactobionate, malate, maleate, mandelate, methylenebis(salicylate), mucate, methanesulfonate, napadisylate, napsylate, pamoate, pantothenate, pectinate, phosphate/diphosphate, polygalacturonate, propionate, salicylate, stearate, succinate, sulfate, tartrate, tosylate and undecanoate. Other anionic species will be apparent to the ordinarily skilled chemist.

The compounds of the invention can be included in a variety of pharmaceutical preparations. Compositions for injection, the preferred route of delivery, may be prepared in unit dosage form in ampoules or in multidose containers. The compositions may take such forms as suspensions, solutions or emulsions, oily or aqueous in nature, and may contain various formulating agents, such as diluents, buffers, preservatives and the like. Hence, the compound is present in the pharmaceutical composition in combination with these pharmaceutically acceptable carriers.

Alternatively, the active ingredient may be in the form of a powder, which can be reconstituted with a suitable carrier such as sterile water, normal saline and the like at the time of administration. The powder can be in lyophillized or non-lyophillized form.

Oral compositions are typically in the form of tablets, capsules, solutions or suspensions. Such compositions may likewise be packaged in unit dose or multidose containers. In these oral compositions, the pharmaceutically acceptable carriers may be comprised of diluents, tabletting and granulating aids, lubricants, disintegrants, buffers, sweeteners, preservatives and the like.

Topical applications may be formulated with a pharmaceutically acceptable carrier in the form of hydrophobic or hydrophilic ointments, creams, lotions, solutions, paints or powders.

The dosage to be administered depends to a large extent upon the condition and size of the subject being treated as well as the route and frequency of administration. The parenteral route (by injection) is preferred for generalized infections. Such matters, however, are typically left to the discretion of the clinician according to principles of treatment well known in the antibacterial arts.

The compositions for human delivery per unit dosage, whether liquid or solid, may contain from about 0.01 % to about 99% of active material, the preferred range being from about 10-60%. The composition will generally contain from about 15 mg to about 2000 mg of the active ingredient; however, in general, it is preferable to employ a dosage amount in the range of from about 250 mg to 1000 mg. In parenteral administration, the unit dosage is usually the compound I in a sterile water or saline solution or in the form of a soluble powder intended for solution.

The preferred method of administration of the compound of formula I is parenterally by intravenous (i.v.) infusion. Alternatively, the compound may be administered intramuscularly (i.m.). For adults, a dose of about 5 to about 50 mg of the formula I antibacterial compound per kg of body weight is administered from 1 to 6 times per day. The preferred dosage ranges from about 250 mg to 1000 mg of the compound given one (qd), two (b.i.d.) three (t.i.d.) or four (q.i.d.) times per day.

More specifically, for mild infections a dose of about 250 mg one to four times daily is preferred. For moderate infections against highly susceptible gram positive organisms a dose of about 500 mg qd. to q.i.d. is preferred. For severe, life-threatening infections against organisms at the upper limits of sensitivity to the antibiotic, a dose of about 1000-2000 mg three to six times daily is preferred.

For children, a dose of 5-25 mg/kg of body weight given 2,3, or 4 times per day is preferred.

The compound of formula I is of the broad class known as carbapenems. Naturally occuring carbapenems are susceptible to attack by a renal enzyme known as dehydropeptidase (DHP). This attack or degradation may reduce the efficacy of the carbapenem antibacterial agent. The compound of the present invention is significantly less subject to such attack, and therefore may not require the use of a DHP inhibitor. However, such use is optional and contemplated to be part of the present invention. Inhibitors of DHP and their use with carbapenem antibacterial agents are disclosed in European Patent Applications No. 79102616.4, filed July 24, 1979 (Patent No. 0 007 614); and No.

82107174.3, filed August 9, 1982 (Publication No. 0 072 014)].

The compound of the present invention may, where DHP inhibition is desired or necessary, be combined or used with the appropriate DHP inhibitor as described in the aforesaid patents and published application. The cited European Patent Application defines the procedure for determining DHP susceptibility of the present carbapenems and disclose suitable inhibitors, combination compositions and methods of treatment.

A preferred weight ratio of Formula I compound: DHP inhibitor in the combination compositions is about 1 :1. A preferred DHP inhibitor is 7-(L-2-amino-2-carboxyethylthio)-2-(2,2-dimethyl cyclopropanecarboxamide)-2-heptenoic acid or a useful salt thereof.

The compounds of the present invention are active against various gram-positive and to a lesser extent gram-negative bacteria, and accordingly find utility in human and veterinary medicine. The compounds are potent anti-MRSA/MRCNS agents.

The invention is further described in connection with the following non-limiting examples.

EXAMPLE 1

To a stirred solution of 1 ,4-diazabicyclo[2.2.2]octane ( 17.93 g, 0.1599 mol) in 200 ml of acetonitrile at 0 ºC was added dropwise a solution of 2-bromoacetamide (20.05 g, 0.1453 mol) in 200 ml of acetonitrile during 25 min. A precipitate began depositing several minutes into the addition. The milky-white reaction mixture was stirred and allowed to warm to room temperature overnight.

After 20 h, the white solid was isolated by filtration, washing with ~200 ml of acetonitrile. Drying in vacuo gave 33.37 g of white solid. This material was combined with product from a previous preparation giving a total of 37.93 g for recrystallization. The solid was dissolved in ~230 ml of boiling ethanol, filtered while hot, and allowed to cool slowly overnight. The solution was seeded while still very warm. The mother liquors were decanted from the dense crystalline mass and the crystals were washed three times with cold ethanol (~50 ml total). Drying in vacuo gave 30.47 g of 1 as large colorless hygroscopic prisms, mp 194- 198ºC. ¹H-NMR (400 Mz, 2:1 D₂O/CD₃CN): δ 3.48 (t, J = 7.5 Hz, 6H), 3.90 (t, J = 7.5 Hz, 6H), 4.28 (s, 2H).

EXAMPLE 2

To a stirred solution of the bromide salt 1 (30.43 g, 0.1217 mmol) in methanol at room temperature was added dropwise a solution of silver trifluoromethanesulfonate (29.69 g, 0.1 156 mol, 0.95 eq.) in 100 ml of acetonitrile during 20 min. The reaction mixture was protected from light. A precipitate formed immediately and the resulting yellow reaction mixture was stirred for 30 min and then filtered, washing with acetonitrile. The filtrate was evaporated in vacuo to give a white solid which was taken-up in 600 ml of acetonitrile, filtered to remove a small amount of insoluble material and the filtrate evaporated to give 37.1 g of a white solid. The crude solid was recrystallized from ~550 ml of boiling ethanol, allowing the solution to cool to room temperature slowly overnight and seeding while still hot. The mother liquors were decanted from the crystalline mass and the crystals were washed twice with 75 ml of cold ethanol. Drying in vacuo yielded 32.93 g of 2 as white flakes, mp 174-176 ºC.

¹H-NMR (400 Mz, d₆-acetone): δ 3.29 (t, J = 7.6 Hz, 6H), 3.77

(t, J = 7.6 Hz, 6H), 4.20 (s, 2H), 7.1 (bs, 1H), 7.6 (bs, 1H). EXAMPLE 3

To a stirred solution of p-nitrobenzyl (5R,6S)-2-[7-hydroxymethyl-fluoren-9-on-3-yl]-6-(1R-triethylsilyloxyethyl)-carbapen-2-em-3-carboxylate (25.878 g; 39.52 mmol) in anhydrous dichloromethane (395 mL) cooled to 0 °C under an atmosphere of N₂ was added triethylamine (9.36 mL; 67.18 mmol; 1.7 equiv). Upon completion of the addition, methanesulfonyl chloride (3.67 mL; 47.42 mmol; 1.2 equiv) was added dropwise. Stirring was continued for 10 min at 0 °C at which time TLC (SiO₂, LI EA/Hexane) indicated complete conversion of alcohol to mesylate. The reaction mixture was poured into Et₂O/EtOAc (2:1 ) and washed with water. The aqueous layer was removed and the organic layer was washed with saturated NaHCO₃ solution (2x), water (1x), and brine (2x). The organic layer was dried over granular Na₂SO₄, filtered and the solvent removed in vacuo. The resulting foam was dissolved in acetone (395 mL). To this stirred solution was added NaI (7.1 g; 47.42 mmol; 1.2 equiv) and the reaction mixture was protected from light. Stirring was continued for 45 min at ambient temperature at which time TLC ( SiO₂, 1 :1 EA/Hexane) indicated complete conversion of mesylate to iodide. The reaction mixture was poured into Et₂O/EtOAc (1 : 1 ) and washed with Na₂S₂O₃ solution (2x), water (lx), Na₂S₂O₃ solution (1x) and brine (3x). The organic layer was dried over granular Na₂SO₄, filtered and the solvent removed in vacuo. The residual was lyophilized from benzene to provide iodide 1 as a bright yellow dense lyophilizate (28.71 g;).

¹H-NMR (400 MHz, CDCl₃): δ 0.60 (q, J = 7.9 Hz, 6H), 0.94 (t, J = 7.9 Hz, 9H), 1.29 (d, J = 6.2 Hz, 3H), 3.21 (dd, J = 18.3, 10.0 Hz, 1H), 3.27 (dd, J = 6.0, 2.9 Hz, 1 H), 3.35 (dd, J = 18.3, 8.9 Hz, 1H), 4.27 (m, 1H), 4.33 (ddd, J = 10.0, 8.9, 2.9 Hz, 1H), 4.44 (s, 2H), 5.17 (d, J = 13.6 Hz, 1H), 5.34 (d, J = 13.6 Hz, 1H), 7.22 (dd, J = 7.7, 1.4 Hz, 1H), 7.29 (d, J = 7.7 Hz, 1H), 7.42 (s, 1 H), 7.45 (dd, partially obscured, J = 7.6, 1.7 Hz, 1H), 7.47 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 7.7 Hz, 1H), 7.63 (d, J = 1.5 Hz. 1H), 8.07 (d, J = 8.7 Hz, 2H).

EXAMPLE 4

To a stirred solution of iodide 3 (23.335 g; 30.52 mmol) in anhydrous acetonitrile (280 mL) under an atmosphere of N₂ and protected from light was added the DABCO salt 36 (9.45 g; 29.60 mmol; 0.97 equiv). An addition funnel was charged with a solution of silver trifluoromethanesulfonate (7.53 g; 29.30 mmol; 0.96 equiv) in acetonitrile (20 mL). Upon dissolution of 36 (~2-3 min), the silver trifluoromethanesulfonate solution was added dropwise over a 10 min period. The reaction mixture was stirred for 50 min at room

temperature during which time silver iodide precipitated from the solution. TLC (SiO₂, 1 :1 EtOAc/Hexane) indicated complete

consumption of iodide 37. The solids were removed by filtration through a pad of Celite^®, followed by washing the pad with acetonitrile (20 mL). After removing the acetonitrile in vacuo, the resulting oil was dissolved in acetone (225 mL) and again filtered through a pad of Celite^®. The volume of acetone was adjusted to 192 mL, and hexane (1 13 mL) was added. The solution was placed in an ice bath for one hour and the resulting precipitate was collected by suction filtration. The precipitate was washed well with a cold solution of 2:1 acetone/hexane, followed by Et₂O. The yellow solid was dried under high vacuum overnight providing 23.0 g of 4. The mother liquor was reprocessed to provide an additional 3.9 g of 4 .

¹H-NMR (400 MHz, d₆-acetone): δ 0.65 (q, J = 7.9 Hz, 6H), 0.98 (t, J = 7.9 Hz, 9H), 1.28 (d, J = 6.2 Hz, 3H), 3.33 (dd, J = 18.2, 10.2 Hz, 1H), 3.54 (dd, J = 4.8, 3.1 Hz, 1H), 3.67 (dd, J = 18.2, 8.7 Hz, 1 H), 4.35 (m, 1H), 4.42 (ddd, partially obscured, J = 10.2, 8.7, 1.5 Hz, 1H), 4.4-4.5 (m, 6H), 4.55-4.65 (m, 6H), 4.66 (s, 2H), 5.19 (s, 2H), 5.23 (d, J = 13.7 Hz, 1H), 5.40 (d, J = 13.7 Hz, 1H), 7.27 (bs, 1H), 7.50 (dd, J =7.7, 1.4 Hz, 1H), 7.60 (d, J = 8.8 Hz, 2H), 7.63 (d, J = 7.7 Hz, 1H), 7.68 (bs, 1H), 7.87 (d, partially obscured, 1H), 7.89 (s, 2H), 7.95 (dd, J =7.7, 1.6 Hz, 1H), 8.06 (d, J = 8.8 Hz, 2H).

EXAMPLE 4A

A solution of p-nitrobenzyl (5R,6S)-2-[7-hydroxymethyl-fluoren-9-on-3-yl]-6-(1R-triethylsilyloxyethyl)-carbapen-2-em-3-carboxylate (1.507 g, 2.302 mmol) in 20 ml of dichloromethane was cooled to -72 °C and 2,4,6-collidine (0.37 ml, 2.8 mmol, 1.2 eq.) was added followed by trifluoromethanesulfonic anhydride (0.425 ml, 2.5 mmol, 1.1 eq.). After 20 min, a solution of 1,4-diazabicyclo[2.2.2] octane (0.568 g, 5.06 mmol, 2.2 eq.) in 2.0 ml of dichloromethane was added dropwise. The reaction was allowed to proceed for 35 min at -72°C, at which point TLC (SiO₂) showed that no starting material remained. The reaction mixture was diluted into ethyl acetate, washed twice with water, and dried over sodium sulfate. Evaporation in vacuo gave 2.358 g of a yellow foam. The crude product was dissolved in ~8 ml of dichloromethane, transferred to a centrifuge tube, and precipitated by the addition of ~30 ml of diethyl ether with stirring. The solid was isolated by centrifugation and the precipitation process was repeated once. The solid was then transferred to a flask by dissolving in dichloromethane and evaporating in vacuo to provide 1.796 g of 4A as a dry red-orange foam.

¹H-NMR (400 MHz, CDCl₃): δ 0.61 (q, J = 7.8 Hz, 6H), 0.95 (t,

J = 7.8 Hz, 9H), 1.29 (d, J = 6.2 Hz, 3H), 3.10-3.25 (m, 7H), 3.34 (dd, J = 5.6, 2.9 Hz, 1H), 3.39 (dd, partially obscured, 1H), 3.45-3.55 (m, 6H), 4.28 (m. 1H), 4.36 (ddd, J = 9.5, 9.2, 2.9 Hz, 1H), 4.63 (s, 2H), 5.12 (d, J = 13.5 Hz, 1H), 5.3 (d, J = 13.5 Hz, 1H), 7.18 (dd, J = 7.7, 1.2 Hz, 1H), 7.3 (s, 1H), 7.34 (d, J = 7.9 Hz, 2H), 7.34 (d, J = 7.5 Hz, 1H), 7.46 (d, J = 7.7 Hz, 1H), 7.50 (s, 1H), 7.68 (d, J = 7.5Hz, 1H), 7.92 (d, J = 7.9 Hz. 2H).

EXAMPLE 4B

To a stirred solution of 4A (106.4 mg; 0.1 19 mmol) in CH₃CN (1.2 mL) was added as a solid 2-bromoacetamide (16.4 mg; 0.1 19 mmol; 1.0 equiv). After 15 min a 1.0 M solution of silver trifluoromethanesulfonate in CH₃CN (0.119 mL; 0.1 19 mmol; 1.0 equiv) was added. After 48 h, the reaction mixture was filtered through a 0.45 μm filter disc to remove the dark green solid that had formed and evaporated in vacuo to give a yellow foam. The residue was dissolved in a minimum amount of acetone and transferred to a centrifuge tube. Ethyl ether was added to precipitate the products and the suspension was centrifuged. After removal of the supernatant, the yellow precipitate was redissolved in acetone and the process was repeated. The precipitate was again dissolved in acetone and filtered through a 0.45 μm filter disc. Evaporation in vacuo gave 103 mg of product as a yellow solid. RP HPLC (LiChrospher 100, RP-18; 90: 10 CH₃CN/0.10 M NH₄CI) showed the product to be approximately a 1 :1 mixture of 4 and des-silyl 4.

EXAMPLE 5

To a stirred solution of the carbapenem 4 (11.965 g; 10.82 mmol) in 2:1 THF/H₂O (309 mL) cooled to 0 °C (initial pH=5.6) was added 1N HCl (8.0 mL; 0.73 equiv) by dropwise addition. The pH of the resulting solution was 2.25. The TES group was completely removed after 50 min at 0 °C as judged by RP HPLC (LiChrospher 100, RP-18; 86:14 MeCN/0.05N NH₄CI). The pH of the solution was adjusted to 6.8 by addition of 1M NaHCO₃ solution (11.0 mL; 1.01 equiv) followed by 0.5M pH 6.8 MOPS buffer (103 mL). Decolorizing charcoal (1.2 g) was added, the ice bath was removed, and the reaction mixture was rapidly stirred for 15 min. After addition of 5% Rh/C catalyst (600 mg), the reaction vessel was evacuated and purged with H₂ (10x) and the reaction was stirred at top speed under an atmosphere of H₂. After 15 min, additional 5% Rh/C catalyst (600 mg) was added. The reaction was judged to be complete after 36 min by RP HPLC (LiChrospher 100, RP-18; 44:56 MeCN/0.02N NH₄CI). The reaction vessel was purged with nitrogen and the catalyst was filtered off through a pad of Celite^®, washing the pad well with water. The

filtrate was extracted thoroughly with EtOAc, filtered through a pad of Celite^®, and then frozen and lyophilized overnight (approximately 20 h). The crude lyophilizate (19.8 g) was taken-up in aqueous NaCl solution and purified by MPLC through a column packed with Amberchrom^® CG-1000sd resin to give, after lyophilization, 5.75 g of 5 as a fluffy orange, amorphous solid.

UV (H₂O): λ_max=306, 374 nm.

¹H-NMR (400 MHz, 2:1 D₂O/CD₃CN): δ 1.60 (d, J = 6.5 Hz, 3H), 3.35 (dd, J = 16.5, 9.8 Hz, 1H), 3.74 (dd, J = 16.5, 8.3 Hz, 1H), 3.80 (dd, J = 5.7, 2.9 Hz, 1H), 4.25-4.35 (m, 6H), 4.55-4.65 (m, 6H), 4.55-4.65 (2H, partially obscured), 4.63 (s, 2H), 5.06 (AB_q, J_AB = 12 Hz, Δʋ_AB = 15 Hz, 2H), 7.58 (dd, J = 7.8 Hz, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.93 (s, 1H), 7.98 (s, 1H), 8.02 (dd, J = 7.8, 1.1 Hz, 1H), 8.10 (d, J = 7.8 Hz, 1H). EXAMPLE 6

Compound 6A (10.0 g, 25.6 mmol) was dissolved in

CH₂Cl₂ (51 mL) and rhodium(II) octanoate (50 mg) was added. The reaction mixture was heated at reflux for 3 h. Additional CH₂Cl₂ (77 mL) was added and the reaction was cooled to -78 °C. Triethylamine (3.75 mL, 26.9 mmol, 1.05 equiv) was then added dropwise over several minutes and the reaction was stirred for 15 min before trifluoromethanesulfonic anhydride (4.52 mL, 26.9 mmol, 1.05 equiv) was added slowly dropwise. The reaction was stirred an additional 15 min before adding triethylamine (3.93 mL, 28.2 mmol, 1.1 equiv) followed by the addition of triethylsilyl trifluoromethanesulfonate (6.36 mL, 28.2 mmol, 1.1 equiv), both added slowly dropwise. This mixture was then stirred for 75 min at -78 °C. The fluorenone boronic acid 6C (7.15 g, 28.2 mmol, 1.1 equiv) was dissolved in 64 mL of 1-methyl-2-pyrrolidinone and added to the reaction vessel via cannula. Tris(dibenzylideneacetone)

dipalladium-chloroform catalyst (531 mg, 0.51 mmol, 0.02 equiv) was then added as a solid. A 6N aqueous KOH solution (12.8 mL, 76.8 mmol) was added last. The ice bath was removed and the reaction vessel warmed briefly using a warm water bath before being placed in an oil bath set at 50 °C. After approximately 45-50 min the enol triflate intermediate 40B was completely consumed according to TLC (SiO₂, 1 :1 EtOAc/Hexanes). The contents of the reaction vessel were poured into Et₂O and washed with saturated NaHCO₃ solution (2x), water/brine mixture [3: 1 ] (5x), and finally brine (2x). The organic layer was treated with decolorizing charcoal for approximately 5 min before MgSO₄ was added. Filtration and removal of the solvent in vacuo afforded a wine-red colored foam which was purified via SiO₂ Flash Chromatography (EtOAc/Hexanes) to provide 10.36 g of 6.

¹H NMR (400 MHz , CDCl₃) δ 0.61 (q, J = 8.05 Hz, 6H), 0.95 (t, J = 7.9 Hz, 9H), 1.1 1 (d, J = 7.3 Hz, 6H), 1.28 (d, J = 6.2 Hz, 6H), 1.92 (t, J = 6.0 Hz, 1H), 3.36 (dd, J = 5.9, 3.1 Hz, 1H), 3.40-3.47 (m, 1H), 4.25-4.32 (m, 1H), 4.39 (dd, J = 10.2, 3.1 Hz, 1H), 4.70 (d, J = 5.5 Hz, 2H), 5.17 (AB_q, J_AB = 13.5 Hz, Δʋ_AB = 33.5 Hz, 2H), 7.16 (dd, J = 7.6, 1.4 Hz, 1H), 7.28-7.35 (complex m, 4H), 7.45 (dd, J = 7.7, 1.5 Hz, 1H), 7.6-7.64 (m, 2H), 7.97 (d, J = 8.8 Hz, 2H).

EXAMPLE 7

A solution of the fluorenone-carbinol 6 (20.070 g, 30.008 mmol) in 300 ml of dichloromethane was cooled to -55 °C and triethylamine (6.0 ml, 43 mmol, 1.4 eq.) was added followed by the dropwise addition of methanesulfonyl chloride (2.8 ml, 36 mmol, 1.2 eq.) during several minutes. The reaction temperature was allowed to rise to -35 °C during 55 min at which point TLC (SiO₂, 1:4

EtOAc/CH₂Cl₂) showed no remaining starting material. The reaction mixture was diluted into 1 :1 ethyl acetate-diethyl ether and washed successively with water, sat. aqueous ammonium chloride solution, water, and brine. Drying over sodium sulfate, filtration, and evaporation gave the crude mesylate intermediate as a yellow foam which was used immediately without purification. The crude mesylate was dissolved in 300 ml of acetone, cooled to 0 °C, and sodium iodide (9.00 g, 60.0 mmol, 2.0 eq.) was added in one portion with stirring. The reaction mixture was stirred in the dark at 0 °C for 1 h and then the ice bath was replaced with a 10 °C bath and the stirring was continued for an additional 1 h. At this point TLC (1 :1 EtOAc/hexane) showed the reaction to be essentially complete with only a trace of mesylate remaining. The cooling bath was removed and the reaction mixture was stirred for 15 min more and then diluted into 1 :1 ethyl acetate-diethyl ether. The organic solution was washed with dilute sodium chloride solution (2x), 5% sodium thiosulfate solution/brine (3:1), dilute sodium chloride solution, and brine. The organic layer was dried over sodium sulfate and evaporated in vacuo leaving a slightly gummy yellow solid. The crude iodide was dissolved in 150 ml of benzene and lyophilized to yield 22.720 g of 7 as a fluffy yellow solid which was used without purification.

¹H-NMR (400 MHz, CDCl₃): δ 0.61 (q, J = 7.8 Hz, 6H), 0.95 (t, J = 7.8 Hz, 9H), 1.10 (d, J = 7.3 Hz, 3H), 1.28, (d, J = 6.2 Hz, 3H), 3.36 (dd, J = 5.8, 3.2 Hz, 1H), 3.44 (m, 1H), 4.30 (m, 1H), 4.40 (dd, J = 10.3, 3.2 Hz, 1H), 4.45 (s, 3H), 5.13 (d, J = 13.5 Hz, 1H), 5.27 (d, J = 13.5 Hz, 1H), 7.20 (dd, J = 7.6, 1.4 Hz, 1H), 7.32 (d, J = 7.5 Hz, 1H), 7.40 (s, 1H), 7.43 (d, J = 8.8 Hz, 2H), 7.47 (dd, J = 7.7, 1.8 Hz, 1H), 7.63 (d, J = 7.7 Hz, 1H), 7.65 (d, J = 1.4 Hz, 1H), 8.06 (d, J = 8.8 Hz, 2H).

EXAMPLE 8

The iodide 7 (12.547 g, 16.112 mmol) was dissolved in 290 ml of acetonitrile and 30 ml of tetrahydrofuran with stirring at room temperature. The ammonium trifluoromethanesulfonate salt 2 (5.145 g, 16.11 mmol) was added in one portion as a solid and dissolved within several minutes. A solution of silver trifluoromethanesulfonate in acetonitrile (0.964 M, 16.0 ml, 15.4 mmol, 0.957 eq.) was added slowly dropwise with stirring in the dark during 40 min. A precipitate began depositing immediately, and after the addition was complete the reaction mixture was stirred for an additional 35 min. Methanol (30 ml) was added and the reaction mixture was filtered through a pad of Celite^®, washing with acetonitrile. The solution was rotary evaporated to low volume and the resulting slurry was dissolved in ~175 ml of 9: 1 acetone-methanol. The slightly hazy solution was filtered through a pad of Celite^® washing with additional 9:1 acetone-methanol. The total volume of the filtrate was ~250 ml of clear orange solution. The solution was vigorously stirred while 250 ml of diethyl ether was added relatively rapidly from an addition funnel (addition time ~10 min).

Precipitation of the product began after ~70 ml of the diethyl ether had been added. After the addition was complete, the mixture was stirred for 5 min more, and then suction filtered through a coarse filter funnel, washing with 300 ml of 2: 1 diethyl ether-acetone followed by 300 ml of diethyl ether. The solid was air-dried on the funnel and then in a vacuum desiccator overnight to yield 15.590 g of 8 as a yellow solid. ¹ H-NMR (400 MHz, d₆-acetone): δ 0.66 (q, J = 8.0 Hz, 6H), 0.99 (t, J = 8.0 Hz, 9H), 1.15 (d, J = 7.4 Hz, 3H), 1.28 (d, J = 6.2 Hz, 3H), 3.54 (dd, J = 4.7, 3.4 Hz, 1H), 3.68 (m, 1H), 4.37 (m, 1H), 4.44-4.51 (m, 7H), 4.55-4.62 (m, 6H), 4.67 (s, 2H), 5.18 (d, J = 13.6 Hz, 1H), 5.20 (s, 2H), 5.29 (d, J = 13.6 Hz, 1H), 7.26 (bs, 1H), 7.50 (dd, J = 7.6, 1.4 Hz, 1H), 7.55 (d, J = 8.8 Hz, 2H), 7.66 (d, J = 7.7 Hz, 1H), 7.68 (bs, 1H), 7.86 (s, 1H), 7.89 (d, J = 7.3 Hz, 1H), 7.89 (d, J = 1.3 Hz, 1H), 7.96 (dd, J = 7.9, 1.5 Hz, 1H). 8.02 (d, J = 8.8 Hz, 2H).

EXAMPLE 9

Carbapenem 8 (4.50g; 4.02 mmol) was dissolved in 120 mL of 2:1 THF/H₂O and cooled to 0 ºC. The pH of this solution, which began at 4.86, was adjusted to 2.30 using 1N HCl (3.40 mL; 3.40 mmol; 0.85 equiv). The disappearance of the TES-group was monitored by RP HPLC (LiChrospher 100, RP-18; 85:15 CH₃CN/0.10 M NH₄CI) and the hydrolysis was judged to be complete after 80 min. The reaction mixture was neutralized to pH 7.0 using 1 M NaHCO₃ (3.60 mL; 3.60 mmol; 0.90 equiv), and 40 mL of 0.5 M pH 6.75 MOPS buffer was added. Ten percent w/w of 5% Rh/C (450 mg) was added and the flask was purged 10 times with H₂. The solution was stirred vigorously under balloon pressure of H₂ for 80 min at which time the reaction was judged to be complete by RP HPLC (LiChrospher 100, RP-18; 35:65 CH₃CN/0.10 M NH₄CI). Following removal of the H₂, the reaction mixture was filtered through a pad of Celite^® rinsing with 2:1

H₂O/CH₃CN. The solution was frozen and lyophilized overnight. The crude product was taken-up in 20% aqueous NaCl, filtered through Celite® and purified by MPLC through a column packed with

Amberchrom^® CG-1000sd resin to yield 1.63 l g of 9 as a fluffy orange solid

UV (H₂O): λmax = 368 nm.

¹H-NMR (400 MHz, 2: 1 D₂O/CD₃CN): δ 1.42 (d, J = 7.2Hz, 3H), 1.60 (d, J = 6.2 Hz, 3H), 3.77 (dd, J = 5.9, 2.7 Hz, 1H), 3.85 (m, 1H), 4.23-4.35 (m, 6H), 4.43-4.55 (m, 6H), 4.54 (m, 1H), 4.62 (s, 2H), 4.64 (dd, partially obscured, 1H), 5.05 (s, 2H), 7.79 (d, J = 8.0 Hz, 1H), 7.94 (d, J = 7.7 Hz, 1H), 7.98 (s, 1H), 8.01 (d, J = 7.7 Hz, 1H), 8.1 1 (d, J = 8.0 Hz, 1H), 8.13 (s, 1 H).

A stirring solution of carbinol 6 (381.5 mg) in CH₂Cl₂

(5.7 ml) was cooled to -70°C under N₂. Collidine (90.4 μl) was added dropwise folowed by trifluoromethanesulfonic anhydride (105.5 μl). The solution was stirred for 25 min, and 1,4-diazabicyclo[2.2.2]octane (140.7 mg) was added as a solid. After an additional 35 min of stirring, the reaction mixture was diluted with EtOAc and washed 2 times with H₂O. The organic layer was dried over Na₂SO₄, filtered and

evaporated in vacuo. The residue was dissolved in minimum CH₂Cl₂ and precipitated with Et₂O. The supernatant was removed and the precipitation was repeated. The resulting precipitate was redissolved in CH₂Cl₂ and evaporated in vacuo to give 395.9 mg of 10. ¹ H-NMR (400 MHz, CDCl₃): δ 0.63 (q, J = 8.0 Hz, 6H), 0.96 (t, J = 7.9 Hz, 9H), 1.11 (d, J = 7.3 Hz, 3H), 1.28 (d, J = 6.2 Hz, 3H), 3.20- 3.30 (m, 6H), 3.35-3.45 (m, 2H), 3.45-3.60 (m, 6H), 4.25-4.35 (m, 1H), 4.42 (dd, J = 10.5, 3.2 Hz, 1H), 4.65 (m, 2H), 5.06 (d, J = 13 Hz, 1H), 5.18 (d, J = 13 Hz, 1H), 7.18 (dd, J = 7.7, 1.2 Hz, 1H), 7.22 (d, J = 8.8 Hz, 2H), 7.28 (s,l H), 7.41 (d, J = 7.6 Hz, 1H), 7.56 (d, J = 1.2 Hz, 1H), 7.58 (d, J = 7.8 Hz, 1H), 7.76 (dd, J = 7.7, 1.5 Hz, 1H), 7.84 (d, J = 8.8 Hz, 2H).

EXAMPLE 11

A stirring solution of carbapenem 10 (389.8 mg) in

CH₂Cl₂ (4.3 ml) was cooled to 0°C under N₂. Methyl trifluoro methanesulfonate (48 μl) was added dropwise and the cooling bath was removed after 5 min. The resulting thick slurry was stirred 15 min, dissolved in CH₃CN and evaporated in vacuo to give 434 mg of 11.

¹H-NMR (400 MHz, d₆-acetone): δ 0.66 (q, J = 8 Hz, 6H), 0.99 (t, J = 8 Hz, 9H), 1.15 (d, J = 7.4 Hz, 3H), 1.28 (d, J = 6.2 Hz, 3H), 3.54 (dd, J =

4.7, 3.4 Hz, 1H), 3.66 (s, 3H), 3.64-3.74 (m, 1H), 4.35-4.45 (m, 13H), 4.49 (dd, J = 10.5, 3.3 Hz, 1H), 5.18 (d, J = 13.5 Hz, 1H), 5.18 (s, 2H), 5.29 (d, J = 13.5 Hz, 1H), 7.50 (dd, J = 7.7, 1.4 Hz, 1H), 7.55 (d, J = 8.8 Hz, 2H), 7.65 (d, J = 7.6 Hz, 1H), 7.85-7.90 (m, 3H), 7.94 (dd, J =

7.8, 1.6 Hz, 1H), 8.02 (d, J = 8.8 Hz, 2H).

EXAMPLE 12

Carbapenem 11 (434 mg) was dissolved in a mixture of THF (8 ml) and H₂O (4 ml) and cooled to 0°C under N₂. A 1 N aqueous solution of HCl (330 μl) was added dropwise to the cloudy reaction mixture. After 70 min of stirring, RP HPLC analysis

(LiChrospher 100, RP-18) showed that the hydrolysis was complete, and the solution was neutralized with 1 M NaHCO₃ (350 μl). A solution of 0.5 M MOPS buffer (pH 6.75, 4 ml) was added followed by 5% Rh/C (43 mg). The suspension was purged 10 times with H₂ and allowed to stir under balloon pressure of H₂ for 30 min at which point additional 5% Rh/C (22 mg) was added. After stirring under H₂ for an additional 70 min the reaction was judged to be complete by RP HPLC (LiChrospher 100, RP-18), and activated charcoal (434 mg) was added. The suspension was purged 10 times with N₂, stirred under N₂ for 15 min, and then filtered through a pad of Celite^® which was rinsed with 2: 1 H₂O/CH₃CN. The filtrate was frozen and lyophilized overnight. The crude product was taken-up in 10% aqueous NaCl, filtered through Celite® and purified by MPLC through a column packed with

Amberchrom^® CG-1000sd resin, to give after lyophilization 107 mg of product 12.

UV (H₂O): λmax = 370 nm.

¹H-NMR (400 MHz, 2:1 D₂O/CD₃CN): δ 1.37 (d, J = 7.0 Hz, 3H), 1.55 (d, J = 6.2 Hz, 3H), 3.56 (s, 3H), 3.73 (dd, J = 5.9, 2.8 Hz, 1H), 3.75-3.85 (m, 1H), 4.20 (bs, 12H), 4.47-4.54 (m, 1H), 4.59 (dd, J = 9.5, 2.6 Hz, 1H), 4.99 (s, 2H), 7.73 (dd, J = 7.8, 1.3 Hz, 1H), 7.89 (d, J = 7.8 Hz, 1H), 7.91 (s, 1H), 7.95 (d, J = 8 Hz, 1H), 8.05 (d, J = 7.6 Hz, 1H), 8.06 (s, 1H). EXAMPLE 13

Triethylamine (918 μl) was added to a stirring solution of aniline (600 μl) in methylene chloride (30 ml) at -50 °C. The solution was stirred for 5 minutes, and bromoacetyl chloride (548 μl) was added to give a precipitate. The resulting suspension was allowed to warm to 0 °C resulting in dissolution of the precipitate. The reaction mixture was diluted with methylene chloride and washed two times with water. Drying over sodium sulfate and evaporation under reduced pressure gave 1.32 g of the title compound.

¹H-NMR (400 MHz, CDCl₃): δ 4.02 (s, 2H), 7.15 (t, J= 7.4 Hz, 1H), 7.35 (t, J= 7.5 Hz, 2H), 7.51 (d, J= 7.5 Hz, 2H), 8.10 (bs, 1H).

EXAMPLE 14

To a stirred solution of 1 ,4-diazabicyclo[2.2.2]octane (246 mg) was added solid N-phenylbromoacetamide 13 (426 mg) in one portion to give a pale yellow solution. A 1.0 M solution of silver trifluoromethanesulfonate in acetonitrile (1.89 ml) was added yielding an off-white precipitate which turned grayish brown as it stirred for 1 hour at room temperature. The reaction mixture was filtered through a pad of Celite^® rinsing the filtercake with acetonitrile. Evaporation of the filtrate under reduced pressure gave a yellow foam. The filtration and evaporation were repeated and the resultant foam was dissolved in acetone and ethyl ether was added to give an oil. Centrifugation of this mixture allowed removal of the supernatant. The remaining oil was dissolved in acetone and evaporated under reduced pressure to give 698 mg of the title compound as a foam.

¹H-NMR (400 MHz, acetone-d6): δ 3.32 (t, J= 7.5 Hz, 6H), 3.82 (t, J= 7.5 Hz, 6H), 4.37 (s, 2H), 7.14 (t, J= 7.4 Hz, 1H) 7.34 (t, J= 7.9 Hz, 2H), 7.66 (d, J= 7.6 Hz, 2H). EXAMPLE 15

Iodide 3 (408 mg) was dissolved in acetonitrile (10 ml), and compound 14 (232 mg) was added as a solid which immediately went into solution. A 1.0 M solution of silver trifluoromethanesulfonate in acetonitrile (507 μl) was added to give a precipitate. The heterogeneous mixture was allowed to stir for 1.75 hours. The green - gray precipitate was removed by filtration through a pad of Celite^® which was rinsed with additional acetonitrile. The filtrate was evaporated under reduced pressure and the residue was taken up in minimum methylene chloride and precipitated with ethyl ether. The mixture was centrifuged and the supernatant was removed. The remaining gummy solid was redissolved in methylene chloride and the process was repeated. The gummy solid was redissolved and evaporated under reduced pressure to give 630 mg of the title compound.

¹H-NMR (400 MHz, acetone-d6): δ 0.65 (q, J= 7.9 Hz, 6H), 0.98 (t, J= 7.9 Hz, 9H), 1.28 (d, J= 6.2 Hz, 3H), 3.33 (dd, J= 18.1 Hz, 10.2 Hz, 1H), 3.54 (m, 1H), 3.67 (dd, J= 18.2 Hz, 8.4 Hz, 1H), 4.33-4.36 (m, 1H), 4.37-4.45 (m, 1H), 4.45-4.55 (m, 6H), 4.60-4.70 (m, 6H), 5.22 (s, 2H), 5.32 (AB_q, J_AB= 13.7 Hz, Δʋ_AB= 55.3 Hz, 4H), 7.15 (t, J= 7.4 Hz, 1H), 7.35 (t, J= 8.0 Hz, 2H), 7.50 (d, J= 7.8 Hz, 1H), 7.58-7.67 (m, 5H), 7.85-7.93 (m, 3H), 7.96 (d, J= 7.7 Hz, 1H), 8.06 (d, J= 8.9 Hz, 2H).

EXAMPLE 16

Carbapenem 15 (614 mg) was dissolved in a mixture of THF (10 ml) and H₂O (5 ml) and cooled to 0 °C. A 1 N aqueous solution of HCl (415 μl) was added dropwise to the reaction mixture. After 70 minutes of stirring, the hydrolysis was complete by RP-HPLC (85: 15 acetonitrile/0.10 M ammonium chloride), and the solution was neutralized with 1 M NaHCO₃ (440 μl). A solution of 0.5 M MOPS buffer at pH = 6.75 (5 ml) was added followed by 5% Rh/C (62 mg). The suspension was purged 10 times with H₂ and allowed to stir vigorously under balloon pressure of H₂ for 120 minutes at which point the reaction was judged complete by RP-HPLC (60:40 0.10 M ammonium chloride/acetonitrile), and activated charcoal (615 mg) was added. The suspension was purged 10 times with N₂ and stirred for 10 minutes. It was filtered through a pad of Celite^® which was rinsed with 2: 1 H₂O/CH₃CN. The filtrate was frozen and lyophilized overnight. The crude lyophilizate was taken up in 70:30 H₂O/CH₃CN, filtered through Celite^® and chromatographed by RP-MPLC on RP-18 silica gel (acetonitrile/0.10 M ammonium chloride). The resulting solution was frozen and lyophilized. The lyophilizate was desalted by MPLC using Amberchrom^® CG-1000sd resin and lyophilized to give 48 mg the title compound.

¹H-NMR (400 MHz, 2:1 D₂O/CD₃CN): δ 1.60 (d, J= 6.5 Hz, 3H), 3.37 (dd, J= 16.5 Hz, 10.0 Hz, 1H), 3.70-3.82 (m, 2H), 4.25-4.40 (m, 6H), 4.50-4.65 (m, 8H), 5.08 (m, 2H), 7.58 (m, 2H), 7.74 (t, J= 8.0 Hz, 2H), 7.81 (d, J= 7.6 Hz, 2H), 7.87 (d, J= 8.0 Hz, 1H), 7.95 (s, 1H), 7.99 (s, 1H), 8.04 (d, J= 7.7 Hz, 1H), 8.12 (d, J= 7.7 Hz, 1H). EXAMPLES 17-63

Employing the procedures described herein, additional compounds of the present invention were prepared. These are described in Table I, which additionally includes characterizing data.

EXAMPLES 64-93

Employing the procedures described herein, additional compounds of the present invention may be prepared as described in Tables II and III.

Claims

WHAT IS CLAIMED IS:

A compound represented by the formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R represents H or methyl;

A' and A" independently represent -(CH₂)_n-, wherein n is an integer of from 1 to 4;

X- represents a pharmaceutically acceptable counterion;

M represents a negative charge, H or an ester forming group;

Q' is selected from the group consisting of: a direct bond, -O-, -S-, -C(O)-, -CO(O)-, -OC(O)-, -C(O)N(R^z')-, -N(R^z')C(O)-, -SO₂-, -SO-, -SO₂N(R^z')-, -N(R^z')SO₂-, -OC(O)N(R^z')- and

-N(R^z')C(O)O-;

(a) when Q' represents -S-, -C(O)-, -OC(O)-, -C(O)N(R^z')-, -N(R^z')C(O)-, -N(R^z')SO₂- or -N(R^z')C(O)O-, R^e' represents a member selected from the group consisting of:

C₁ to C₁₀ alkyl (including C_3-10 branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₆ to C₁₄ aryl, unsubstituted or substituted with 1-3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1-3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5-10 atoms, 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups;

(b) when Q' represents -C(O)O-, R^e' represents a member selected from the group consisting of:

C₂ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups; C₈ to C₁₄ aryl, unsubstituted or substituted with 1-3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1-3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5 - 10 atoms, 1 -3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups;

(c) when Q' represents -O-, -SO-, -SO₂-, -SO₂N(R^z')- or -OC(O)N(R^z')-, R^e' is selected from the group consisting of:

C₂ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₆ to C₁₄ aryl, unsubstituted or substituted with 1 -3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1-3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5 - 10 atoms, 1-3 of which are heteroatoms, 0-3 of which are N, and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups; and

(d) when Q' represents a direct bond, R^e' represents a member selected from the group consisting of: C₄ to C₁₀ alkyl (including C_{3- 10} branched and C_{3- 10} cyclic), unsubstituted or substituted with 1 to 3 R^q' groups;

C₆ to C₁₄ aryl, unsubstituted or substituted with 1 -3 R^q' groups; alkaryl or aralkyl, having from 1 to 6 carbon atoms in the alkyl portion thereof, and from 6 to 10 carbon atoms in the aryl portion thereof, said alkyl and aryl portions being unsubstituted or substituted with 1 to 3 R^q' groups; heteroaryl, containing from 5 to 10 atoms, from 1 -3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said heteroaryl group being unsubstituted or substituted with 1 -3 R^q' groups, and alkyl-heteroaryl or heteroaryl-alkyl, the alkyl portion of which having from 1 to 6 carbon atoms, and the heteroaryl portion of which having from 5-10 atoms, 1-3 of which are heteroatoms, 0-3 of which heteroatoms are N and 0-1 of which are O or S, said alkyl and heteroaryl portions thereof being unsubstituted or substituted with 1 to 3 R^q' groups;

R^z is selected from the group consisting of: H, OCH₃, C₁ to C₄ alkyl and C₁ to C₄ alkyl substituted with 1-3 R^q' groups, and each R^q' is independently selected from the group

consisting of: "OH, -OCH₃, -CN, -C(O)NH₂, -C(O)NHCH₃,

-C(O)N(CH₃)₂, -OC(O)NH₂, -CHO, -SO₂NH₂, -SOCH₃, -SO₂CH₃, -F, -CF₃, -C(O)CH₃, -COOCH₃ and -COOM^a (where M^a is H, a negative charge or an alkali metal).

2. A compound in accordance with claim 1 wherein M represents a negative charge.

3. A compound in accordance with claim 1 wherein A' and A" are selected from -(CH₂)_n- wherein n represents 1 or 2.

4. A compound in accordance with claim 2 wherein n equals 1.

5. A compound in accordance with claim 1 wherein R is methyl.

6. A compound in accordance with claim 1 wherein R is hydrogen.

7. A compound in accordance with claim 1 wherein Q' is selected from the group consisting of: a direct bond, -C(O)- and -C(O)N(R^z')- .

8. A compound in accordance with claim 1 wherein R^e' is selected from the group consisting of: C₁ to C₃ alkyl, substituted with 1 R^q' group, and phenyl substituted with 1 R^q' group.

9. A compound in accordance with claim 1 wherein R^z' is selected from the group consisting of: H, C_{1 -4} alkyl and C_{1 -4} alkyl substituted with 1 R^q' group.

10. A compound in accordance with claim 1 wherein R^q' is selected from the group consisting of: OH, -CN, -C(O)NH₂, -CHO, -SO₂NH₂, -SOCH₃, -SO₂CH₃, -F, -CF₃ and -C(O)CH₃.

1 1. A compound in accordance with claim 9 wherein R^q' is selected from the group consisting of: OH, -CN and -C(O)NH₂.

12. A compound in accordance with claim 1 represented by formula la:

wherein R, M, A', A", Q, X and R^e' are as previously defined.

13. A compound in accordance with claim 1 represented by formula Ib:

wherein R, M, A', A", Q, X and R^e' are as previously defined.

14. A compound in accordance with claim 1 represented by formula Ic:

wherein R, M, A', A", Q, X and R^e' are as previously defined.

15. A compound in accordance with claim 13 wherein R and R^a are as set forth in Table I:

and X- represents a pharmaceutically acceptable counterion.

16. A compound in accordance with claim 1 wherein R and R^a are as set forth in Table II:

and X- represents a pharmaceutically acceptable counterion.

17. A compound in accordance with claim 14 wherein R and R^a are as set forth in Table III:

and X- represents a pharmaceutically acceptable counterion.

18. A compound in accordance with claim 12 wherein R and R^a are as set forth in Table IV:

19. A compound in accordance with claim 12 wherein R and R^a are as set forth in Table V:

20. A compound in accordance with claim 1 in the form of a pharmaceutically acceptable salt, wherein X- represents a member selected from the group consisting of: acetate, adipate, aminosalicylate, anhydromethylenecitrate, ascorbate, aspartate, benzoate, benzene-sulfonate, bisulfate, bromide, citrate, camphorate, camphorsulfonate, chloride, digluconate, edetate, edisylate, estolate, ethanesulfonate, fumarate, glucoheptanoate, gluconate, glutamate, glycerophosphate, glycolate, hydroxynaphthoate, 2-hydroxyethanesulfonate, iodide, lactate, lactobionate, malate, maleate, mandelate, methylenebis(salicylate), mucate, methanesulfonate, napadisylate, napsylate, pamoate,

pantothenate, pectinate, phosphate, diphosphate, polygalacturonate, propionate, salicylate, stearate, succinate, sulfate, tartrate, tosylate and undecanoate.

21. A compound in accordance with claim 20 wherein

X- represents chloride.

22. A pharmaceutical composition comprising a

compound in accordance with claim 1 in combination with a

pharmaceutically acceptable carrier.

23. A method of treating a bacterial infection in a mammalian patient, comprising administering to a mammalian patient in need of such treatment an antibacterially effective amount of a

compound as defined in claim 1.