PH26648A - Acyl derivatives - Google Patents

Description

ACYI. DERIVATIVES

The present invention relates to intermediates and antibacterial compounds of the formula . (Ol

Ho Ba HO li is

Rye N

N Ry i COCR wherein R is hydrogen or a carboxylic acid-protecting group; R, is a substituted piperazinium group of the formula of J or ®

CC 5 | rO a0 H CH 05 Cc 3 3 in which Q represents a substituted quinolinyl or naphthyridinyl group and the piperazine nucleus may optionally be substituted with one or more lower alkyl groups; R, is selected from the group consisting of

- 2 2G x hydrogen, lower alkoxy, lower alkylthio and lower alkanoylamino; R, is hydrogen or an acyl group; and m is 0, 1 or 2, but preferably 0; and A is a pharmaceuti- cally acceptable anion; as well as the corresponding readily hydrolyzable esters, pharmaceutically acceptable salts and hydrates of these compounds. )

As used herein, the terms "lower alkyl" and "alkyl" 40 refer to both straight and branched chain saturated hydrocarbon groups having 1 to 8, and preferably 1 to 4, carbon atoms, for example, methyl, ethyl, propyl, isopropyl, tertiary butyl, and the like.

As used herein, the term "lower alkoxy" refers to a straight or branched chain hydrocarbonoxy group wherein the "alkyl" portion is a lower alkyl group as defined above.

Examples include methoxy, ethoxy, propoxy and the like.

The term "halogen", or "halo", used herein refers to all four forms, that is, chloro, bromo, iodo and fluoro, unless specified otherwise. ' The term "acyl" used in conjunction with Ry herein o5 refers to all organic radicals derived from an organic carboxylic acid by removal of the hydroxyl group. Although the group R, may be any one of many acyl radicals, certain acyl groups are preferred, as described below.

Exemplary acyl groups are those groups which have been used in the past to acylate beta-lactam antibiotics, including 6-aminopenicillanic acid and derivatives and 7- aminocephalosporanic acid and derivatives; see, for example,

Cephalosporins and Penicillins, edited by Flynn, Academic

Press (1972). Belgian patent 866,038, published October 17, 1978, Belgian patent 867,994, published December 11, 1978,

United States patent 4,152,432, issued May 1, 1979, United a A66Y

States patent 3,971,778, issued July 27, 1976, and United

States patent 4,173,199, issued October 23, 1979. The portions of these references describing various acyl groups are incorporated herein by reference. The following list of acyl groups is presented to further exemplify the term "acyl", without intending to limit that term to only those groups set forth: (a) Aliphatic acyl groups having the formula ?

Reem C =~ . wherein Re is hydrogen, alkyl, cycloalkyl: alkoxy: alkenyl: cycloalkenyl: cyclohexadienyl: or alkyl or alkenyl substituted with one or more halogen, cyano, nitro, amino, mercapto, alkylthio, or cyanomethylthio groups. : (b) Aromatic acyl groups having the formula

Ry

Re Re i

Ry oo 30 SH—C—-

Reo

Ry

Re Re 0

CHy== 0 ——C—f—

“J {ol /) ? oq -

Ry 0

Ry h i

Ry

A oO

Re ‘ Il § — CH; — C—+

By

R 0

Pe ol - and '

SO,” M'

Ry

Re 0

Re Il

NH

- : SO,” M wherein n is 0, 1, 2 or 3: Re. R.. and Rg each is independently hydrogen, halogen, hydroxyl. nitro, amino, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or aminomethyl: and Roo is amino, acylamino, hydroxyl, a carboxyl salt, protected carboxy such as benzyloxycarbonyl, formyloxy or azido; and M is a cation.

Preferred aromatic acyl groups include those having the formula i

I

I i ’ { Hs —C—

Reo . . and i : Sg Sr

Roo is preferably an amino group, a hydroxy group. or a carboxyl salt or sulfo salt.

Examples of other aromatic acyl groups suitable for the purposes of the present invention are sulfophenylacetyl, hydroxysulfonyloxyphenylacetyl, sulfamoylphenylacetyl, ‘(phenoxycarbonyl)phenylacetyl, (p-tolyloxycarbonyl)phenyl- acetyl, formyloxyphenylacetyl, carboxyphenylacetytl, formylaminophenylacetyl, benzyloxycarbonylphenylacetyl, 2-(N,N-dimethylsulfamoyl)-2-phenylacetyl, etc. (c) Hetercaromatic acyl groups having the formula 0

Ryo = (CH3)g=—C~ i

Reo

2 (» (v A - 6 -

I

Rypy == Q=——CHy—C 0

I

Rygy=—S = CH; —C or 0 O

Rygt—C—~C— wherein n is 0, 1, 2 or 3; Rog is as defined above; and Rio is a substituted or unsubstituted 5-, 6- or . 7-membered heterocyclic ring containing 1, 2. 3 or 4 (preferably 1 or 2) hetero atoms selected from among nitrogen, oxygen and sulfur. Fxemplary heterocyclic rings are thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl, pyrimidinyl and taelrazolytl.

Exemplary substituents are halogen, hydroxyl, nitro, amino, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms.

Preferred heteroaromatic acyl groups include those groups of the above formulas wherein R101 is 2-amino-4- thiazolyl, 2-amino-5-halo-4-thiazolyl, 4-aminopyridin-2-yl, 2-amino-1,3,4-thiadiazol-5-yl, 2-Lhienyl, 2-furanyl, 4-pyridinyl ocr 2,6-dichloro-4-pyridinyl. (d) ([(4-Substituted-2,3-dioxo-1-piperazinyl)cacbonyl] amino acetyl groups having the formula 0 oO il Il /\ 4-C—CH—NH—C—N N— Riz

Ryn y Q

26(-4§ wherein Ri; is alkyl, hydroxyalkyl or an aromatic heterocyclic or carboxylic group, such as those of the formula

Ry ~~ wherein Re. R, and Rg are as previously defined or heteroaromatics as included within the definition of

Rio1¢ and Riso is alkyl, substituted alkyl (wherein the alkyl group is substituted with one or more halogen, - cyano, nitro, amino or mercapto groups), e.g., 41-lower i5 alkyl (preferably ethyl or methyl)--2,3-dioxo-1- piperazinecarbonyl-D-phenylglycyl. (e) (Substituted oxyimino) arylacetyl groups having the formula

Q

I

Ryo wherein R101 is as defined above and R 30 18 hydrogen, lower alkyl and C,-C, cycloalkyl or substituted lower alkyl wherein the alkyl group is substituted with one or more halogen, cyano, nitro, amino, mercapto, lower alkylthio, aromatic group (as defined by Ryp00 carboxyl (including salts thereof), lower alkanoylamino, carbamoyl, lower alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxycarbonyl, hydroxy- alkoxyphosphinyl, dihydroxyphosphinyl, hydroxy(phenyl- methoxy)phosphinyl, di-lower alkoxyphosphinyl substi- tuents, carboxyl lower alkyl or carboxyl-C,-C,- -cycloalkyl.

i , CG - 8 - Jed 2

Examples of the 0 i]

FO ENO A

Ryo grouping are [2-[(chlorocacetyl)amino]l-4-thiazolyl](methoxy- amino)acetyl, (2-amino-4-thiazolyl)(l-methylethoxyimino)- acetyl, (2-amino-4-thiazolyl)(methoxyimino)acetyl, (2-furyl)- (methoxyimino)acetyl. (4-hydroxyphenyl)(methoxyimino)acetyl, (methoxyimino) (phenyl)acetyl, (hydroxyimino)(phenyl)acetyl,. (hydroxyimino)(2-thienyl)acetyl, [[(dichloroacetyl)oxy)imi- no)J(2-thienyl)acetyl, [5-chloro-2-[(chloroacetyl)amino}-4- thiazolyl](methoxyimino)acetyl, (2-amino-5-chloro-4-thia- : zolyl)(methoxyimino)acetyl, [[[l-(l.,1l-dimethylethoxy)car- bonyl)-1-methylethoxy)iminoj-2-sulfoamino-4-thiazolyl)ace- tyl, [[[1-(1l.l-dimethylethoxycarbonyl]}-l-methylethoxylimino]- {{2-(triphenylmethyl)amino}-4-thiazolyl]acetyl, (methoxyimi- no)(2-sulfoamino-4-thiazolyl)acetyl, [(l-methylethoxy)imino]- {2-[(methylsulfonyl)amino}-4-~thiazolyl]acetyl, [(3-methylsul- fonyl)-2([3H]-thiazolimin-4-yl]-{l-(methylethoxy)iminolacetyl, {[2-(chloracetyl)amino]-4-thiazolyl][{[[(4-nitrophenyl)-methox y]Jcarbonyl]methoxyJiminoJacetyl, (2-amino-4-thiazolyl)[(carc- boxymethoxy)iminolacetyl, (2-amino-4-thiazolyl)[l-carboxy-(1l- methylethoxy)iminoJacetyl., (2-amino-4-thiazolyl)([(amino- carbonyl)methoxy]iminojacetyl. (f) (Acylamino) acetyl groups having the formula 0 0

Il Il fe C= CH= NH=— C= Ry

Ryn wherein Bi is as defined above and R.a0 1s

- 9 - > 0 tg

Ry

Re Ry ’ (CHp p= O =~ (where Re. R,. Rg and n are as previously defined), hydrogen. lower alkyl, substituted lower alkyl, amino, alkylamino, dialkylamino, (cyanocalkyl)- amino, hydrazino, alkyl hydrazino, aryl hydrazino or acyl hydrazino.

Preferred (acylamino)acetyl groups of the above formula . include those groups wherein R a0 ig amino, or acylamino.

Also preferred are those groups wherein Ra is phenyl or 2-thienyl. (g) Substituted oxyimino acetyl groups having the formula 0 2 9 +C—C=N—0—C—C—FRw . : Ry Rn wherein Ria and Ry 40 are as defined above, and “Ry, and R,, are independently selected from the group consisting of hydrogen and lower alkyl, or R,, and R,. taken together with the carbon atom to which they are attached form a C,-C, carboxylic ring, for example, cyclopropyl, cyclobutyl or cyclopentyl.

Preferred substituted oxyimino acetyl groups of the ' above formula include those groups wherein Rj40 is hydroxy or amino. Also preferred are those groups wherein Ria is 2-amino-4-thiazolyl.

~ J Ce ( [¥ - 10 - ' (h) [[({3-Substituted-2-oxo-1-imidazolidinyl]carbonyl]}- amino lacetyl groups having the formula 0 0 o

Cc ll Il / \

FOG NH—C—a N— Rg

I

Ryn CH;— CH, wherein Ry is as defined above and Rc is hydrogen, alkylsulfonyl, arylmethyleneamino (i.e... -N=CHR, , wherein Rit is as defined above), COR, (wherein Rig is hydrogen, alkyl or halogen substituted alkyl), aromatic group (as defined by Ria above), . alkyl or substituted alkyl (wherein the alkyl group is substituted with one or more halogen, cyano, nitro, amino or mercapto groups).

Preferred [[[3-substituted-2-oxo-1l-imidazolidinyl]- carbonyl]aminojlacetyl groups of the above formula include those wherein R,,, is phenyl or 2-thienyl. Also preferred are those groups wherein Rg is hydrogen, methylsulfonyl, phenylmethyleneamino or 2-furylmethyleneamino.

As used herein the substituted piperazinium substituent

Ry includes, among others, compounds of the formulae: 0

F FF COzH a oN N N on +H

CH,

2d 8 0

CO,H

CN N

S07

CH, . 15 Q . oy +H NHCH,

CH, . 25

Q

Try ow N

Hi A

CH, ’ 3s

CL DCHUY

0

F CO,H £7 ’ +h on,

CH, 0

F CO,H on N " +h ror

CH, 0

NN on N N +H

CHa

F

0 . Co] (TN NTN aA

CHj

- C13 ote Ap

O 0

F

NH

I

(ON N S aA

CH,

O 0

Lr

NH

/ (oN NTO i N A

CH, . r COM ( \ +1 ON

CHy

LN

0

F COOH

(Cy \ +r Ao §

CH,

By the term "aryl" is meant a substituted or unsubstituted aromatic moiety, such as phenyl, tolyl, xylyl, mesityl, cumenyl, naphthyl, and the like, wherein said aryl group may have 1 to 3 suitable substituents, such as halo (fluoro, chloro, bromo, etc.), hydroxy, and the like.

By the term "lower alkanoyl" or "alkanoyl" as utilized herein is intended a moiety of the formula aa 0 wherein R,¢ is H or Cy to Ce lower alkanoic acid, e.g., acetyl, formyl, propionyl, butyryl and the like.

By the term "substituted phenyl" is meant phenyl mono- or di-substituted by halo(chloro, bromo, fluoro, etc.). lower alkyl, amino, nitro or trifluoromethyl.

By the term "substituted alkyl" is meant a "lower alkyl" moiety substituted by, for example, halo (chloro, fluoro, bromo, etc.), trifluoremethyl, amino, cyano, etc.

By the term "lower alkenyl" is meant straight or branched chain hydrocarbon groups which contain an olefinic double bond having 2 to 6 carbon atoms, i.e., the radical of compounds of the formula CH, 0 wherein n is 2 to 6, e.g.. allyl, vinyl, etc.

By the term "aralkyl" is meant a hydrocarbon group having both aromatic and aliphatic structures. that is, a hydrocarbon group in which a lower alkyl hydrogen atom is substituted by a monocyclic aryl group, e.g., phenyl, tolyl, etc.

The expression 5- or 6- membered heterocyclic ring containing 1-4 hetero atoms selected from the group consisting of O, N and S is intended to represent the following groups: pyridyl, piperidyl, piperidino,

N-oxido-pyridyl, pyrimidyl, piperazinyl, pyrolidinyl, pyridazinyl, N-oxide-pyridazinyl. etc. a S-membered nitrogen-containing hetero ring, e.g. pyrazolyl, imidazolyl , . thiazolyl, 1,2,3-thiadiazolyl. 1,2,4-thiadiazolyl, 1.3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,2,3-oxadiazolyl, 1.2.4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-o0xadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, lH-tetrazolyl, 2H-tetra- ] zolyl, etc., and others. Each of these hetero rings may be further substituted and, as the substituents, there may be mentioned, for example, lower alkyls such as methyl, ethyl, propyl, etc., lower alkoxys such as methoxy, ethoxy, etc., halogens such as chlorine, bromine, etc., halogen substituted alkyls such as trifluoromethyl, trichloroethyl, etc., amino, mercapto, hydroxyl, carbamoyl, or carboxyl group, etc,

By the term "cycloloweralkyl” is meant a 3-6 membered saturated carboxylic moiety, e.g.. cyclopropyl, cyclobutyl, cyclohexyl, etc.

As readily hydrolyzable esters of the compounds of formula I there are to be understood compounds of formula I, the carboxy group(s) of which (i.e., the 2-carboxy group) is/are present in the form of readily hydrolyzable ester ’ groups. Examples of such esters, which can be of the conventional type, are the lower alkanoyloxyalkyl esters (e.g., the acetoxymethyl, pivaloyloxymethyl, l-acetoxymethyl

BN - 16 - 20604 ¢ and l-pivaloyloxyethyl ester), the lower alkoxycarbonyloxy- alkyl esters (e.g., the methoxycarbonyloxymethyl, l-ethoxy- carbonyloxyethyl and l-isopropoxycarbonyloxyethyl ester), the lactonyl esters (e.g., the phthalidyl and thiophthalidyl ester), the lower alkoxymethyl esters (e.g., the methoxy- methyl ester) and the lower alkanoylaminomethyl esters (e.g., the acetamidomethyl ester). Other esters (e.qg.. the benzyl and cyanomethyl esters) can also be used.

Examples of salts of the compounds of formula I are alkali metal salts such as the sodium and potassium salt, the ammonium salt, alkaline earth metal salts such as the calcium salt, salts with organic bases such as salts with . amines (e.g., salts with N-ethyl-piperidine, procaine, dibenzylamine, N,N'-dibenzylethylenediamine, alkylamines or . } dialkylamines) as well as salts with amino acids such as, " for example, salts with arginine or lysine. ‘ The compounds of formula I, when they contain a basic oo functional group such as an amine, also form addition salts with organic or inorganic acids. Examples of such salts are hydrohalides (e.g. hydrochlorides, hydrobromides and hydro- iodides) as well as other mineral acid salts such as sul- phates, nitrates, phosphates and the like, lower alkylsul- phonates and monoarylsulphonates such as ethanesulfonates, toluenesulphonates, benzenesulphonates and the like and also other organic acid salts such as acetates, tartrates, maleates, citrates, benzoates, salicylates, ascorbates and the like.

The. terms "salts" and "pharmaceutically acceptable salts" as employed throughout this disclosure to refer to compounds of formula I also encompass zwitterions and other types of internal salts in which a carboxy group is negatively charged (i.e., lacks a hydrogen atom) and is neutralized by an accompanying positive charge within the : molecule such as the positive charge on the quarternary

A le (4 - 17 - nitrogen atom.

The compounds of formula 1 as well as their salts and readily hydrolyzable esters can be hydrates. The hydration can be effected in the course of the manufacturing process . or can occur gradually as a result of hygroscopic properties of an initially anhydrous product.

A preferred class of compounds are of the formula

NOR;, Ry

Ld i N Ne” A 11 1

I Na on”

Ss oO 0 2

RaoNH

COOH wherein R, and R, are as above, Ryo is hydrogen or an amino-protecting group, for example, trityl or chloroacetyl, and Ry, is hydrogen, lower alkyl or a group of the formula

Rn

Feo

Rx wherein R,, and R,, are as defined above and V is hydroxy or NHR, o where Rio is hydrogen or lower alkyl, amino, alkyl amino, aryl amino or acyl amino.

Still more preferred are compounds of the formula 11 in : which Ro is hydrogen, and Roy is methyl or a group of the formula

- 18 - HG, dr 12 + oon

Rn wherein R,, and R,, are selected from the group consisting of hydrogen and methyl. ee

Preferably, the Cs 7 grouping is in the i : Oo syn-form, e.g., the Z-form.

Another preferred class of compounds are those of the formula

Ra

H ! S och

I N F PR I11 oO Oo CH; ps : COOH wherein Ry and R, are as above. .

Q is a substituted quinolinyl or naphthyridinyl group. preferably of the formula

- 19 - 0G dp 0 : Lo

ND

Z -

Ra wherein Z represents C or N, Rao represents hydrogen, halogen or an oxymethylene (-CH,0-) bridge to the piperazine nucleus to form a fused six-membered ring. . Ry represents hydrogen, lower alkyl, lower alkenyl, Cy-¢, cycloalkyl, halo lower alkyl or mono-, di- and tri-halophenyl;

Ry and Riy when taken together represents lower alkylene of 3-5 carbon atoms, a lower alkylene mono-oxy group of 2-4 carbon atoms, a lower alkylene dioxy group having 1-2 carbon atoms or a group of the formula -OCH,_ N(CH) -: and Ry, represents hydrogen or halogen.

In a preferred embodiment, Z is CR, wherein Rao is hydrogen, chlorine or fluorine, most preferably hydrogen or flourine:; and

Ry, is lower alkyl, most preferably, ethyl or halogen substituted lower alkyl, most preferably, fluorcethyl. or . 30 alternatively, C,-C,-cycloalkyl, most preferably, cyclopropyl: and R,, is chlorine or fluorine, preferably fluorine.

The compounds of Formula I, their pharmaceutically acceptable salts, and esters and hydrates of those compounds can be used as agents to combat bacterial infections (including urinary tract infections and respiratory

2 6p 4p - 20 - infections) in mammalian species, for example, dogs, cats, horses. etc., and humans. These compounds exhibit activity against a broad range of both Gram-negative and

Gram-positive bacteria.

The in vitro activity of the compounds of the present invention as measured by the Minimum Inhibitory

Concentration (MIC) in micrograms/ml utilizing the Broth

Dilution Method against a variety of Gram-positive and

Gram-negative organisms, is as follows:

Compound A: {6R-[6a,7B(Z)))-1-[7-[[[(2-amino-4- thiazolyl) (methoxyimino)acetylJamino]-2-carboxy- . 8-oxo-5-thia-l-azabicyclo{4.2.0]Joct-2-en-3-yl])- methyl]-4-[3-carboxy-6,8-difluoro-1-(2- fluoroethyl)-1,4-dihydro-4-0x0-7-quinolinyl}-1- methylpiperazinium iodide

Compound B: (6R-trans)-4-[(3-carboxy-1-(2-fluoroethyl)-6,8- difluoro-1,4-dihydro-4-oxoquinolin-7-yl1]-1- ((2-carboxy-8-0x0-7-[(phenoxyacetyl)amino]-5- thia--l-azabicyclo-[4.2.0]oct-2-en-3-yl]lmethyl]- l-methyl-piperazinium iodide

Co o5 Compound C: (6R-[6a,7B(Z)1)-1-[[(7-[([(2~amino-4- thiazolyl)[l-(l-carboxy-l1-methyl)ethoxy]- imino l}acetyl]amino)-8-0oxo-5-thia-1-azabicyclo [4.2.0]oct-2-en-3-yl]methyl)}-4-[3-carboxy-1- (2-fluoroethyl)-6,8-difluoro-1,4-dihydro-4- oxoquinolin-7-ylj-l-methylpiperazinium hydroxide inner salt monosodium salt

Compound D: [6R-[6a,7B(Z)])]1-1-[[7-[[(2-amino-4- thiazolyl)(methoxyimino)acetyllamino]-2-carboxy- 8-0x0-5-thia-l-azabicyclof4.2.0)-oct-2-en-3-yl] methyll-4-(3-cacrboxy-1l-ethyl-6-fluoro-1,4-

206 00¢ - 21 - dihydro-4-oxo-7-quinolinyl)-l-methylpipera- zinium iodide

Compound E: (6R-trans)-4-[3-carboxy-6,8-difluoro-1-(2- fluoroethyl)-1,4-dihydro-7-quinolinyl])-1-[[2- carboxy-7-(formylamino)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-en-3-yl]lmethyl])-1- methylpiperazinium trifluoroacetate salt

Table 1

In Vitro MIC (ug/ml), Broth Dilution Method . Compounds 156

Culture A B C D E

E. coli ATCC 25922 0.25 0.5 0.25 0.5 0.5 og E. coli TEM 1 0.25 0.5 0.5 0.5 0.5 .E. cloacae 5699 0.5 -. 0.5. ..0.5 _ . Ll... n.0.5 ..

E. cloacae P99 0.5 0.5 2 1 0.5

S. marcescens 1071 0.5 0.5 0.5 0.5 L : P. aeruginosa 8710 8 8 8 8 8

P. aeruginosa 18 S/H 8 4 8 32 8

E. faecalis ATCC 29212 64 32 32 32 32

S. pneumoniae 6301 0.063 0.25 1 0.063 1

S. aureus 1059B 2 0.5 8 2 0.5

S. aureus 95 4 1 4 4 1 } 30 S. aureus ATCC 29213 4 0.5 4 1 0.5

For combatting bacterial infections in mammals, a compound of this invention (more precisely, a compound of formula I where R is hydrogen or a corresponding hydrolyzable ester or pharmaceutically acceptable salt or hydrate) can be administered to a mammal in an amount of about 5 mg/kg/day to about 500 mg/kg/day. preferably about

10 mg/kg/day to 100 mg/kg/day, most preferably about 10 mg/kg/day to about 55 mg/kg/day.

Modes of administration which have been used in the past to deliver penicillins and cephalosporins to the site of the infection are also contemplated for use with the compounds of the present invention. By way of illustration, such methods of administration include parenteral, e.g., intravenous or intramuscular, and enteral, e.g., as a suppository. . The following reaction schemes set forth the methods and intermediates useful in producing the end products of - formula I. Unless otherwise noted, R,. R, and Q are as breviously defined.

The compound of formula IV, referred to in the Schemes, . is a substituted piperazine of the formula

Q

J oN

N Vv : cy” | : in which the piperazine nucleus may be substituted with one or more lower alkyl groups of 1-8, preferably 1-4 carbon atoms. It shall be understood that in these reaction schemes substituted piperazines of the structure vs

A

. N 1

CHy 35 . may similarly be used as starting materials in place of IV.

Cae ACG

In the following reaction sequences, where a substituent group is present which may be attacked during the reaction it should be in protected form, utilizing well known protecting groups. For example, amino groups may be

Protected with easily removable protective groups employed in peptide chemistry, such as a triphenylmethyl group.

As ester protecting groups one may utilize an ester form which can be easily converted into a free carboxyl group under mild conditions. The ester protecting group can be, for example, t-butyl, p-nitrobenzyl, allyl, etc. Also suitable are trimethylsilyl esters. . Scheme 1

R Q

RyNH 2 _S RyNH Ps ANT

IT O._ CH, 1) MSTFA pd NS —

CO,H © 2) ISiMe, CO,H CH, I° : in 3) IV + MSTFA IA 4) MeOH

Rj Q

RyNH YS _S 7 oy

NA or:

COs CH,

IB

- 24 - S6od¥

Scheme 1

The compound of formula III is initially protected by : reaction with a trimethylsilating agent such as

N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) in an inert solvent such as chloroform or methylene chloride. The resulting trimethylsilyl ester, in which all potentially reactive sites, such as amino, hydroxy. and carboxylic acid functions, are protected by trimethylsilyl groups, is then subjected to a reaction with iodotrimethylsilane at about 0°C to room temperature, over a period of about twenty minutes to about two hours. The reaction mixture is then concentrated to dryness under reduced pressure, and the . residue is dissolved in a suitable non-hydroxylic solvent such as acetonitrile. A small amount of tetrahydrofuran (THF) is introduced to decompose any residual traces of iodotrimethylsilane. The resulting protected iodo intermediate is then further reacted in situ with a protected form of the piperazine derivative IV which is 50 obtained from IV by treatment with a trimethylsilating agent such as MSTFA in a compatible solvent such as acetonitrile.

The quaternization reaction is conducted at about room temperature over a period of about 30 minutes to 24 hours, ‘preferably about 2 hours. Addition of a hydroxylic solvent such as methanol, with ice-cooling, then causes solvolysis of the trimethylsilyl protecting groups, and the quaternary iodide of formula IA precipitates. Further treatment underc aqueous conditions with a base such as sodium bicarbonate or sodium hydroxide, or with sodium phosphate buffer, converts

IA to the zwitterionic form IB in which, depending upon the amount of base added, or the pH of the buffer, other acidic functions in the R, and Q substituents can be converted to salts.

Scheme 11 oo

XD) Ra Q

N 0. CH; N ‘NM 0 7 hd 2)15iMe, o 7

CO,H © CO,H CH, I’ 3) IV + MSTFA

V , IA : 4) Acylation . 5) MeOH , R Q 1) Deprotection RyNH :2 Ss NT if necessary I . oN NA 2) Aqueous base or ] butfer COs CH,

IB

Scheme 11 *

Starting material of structure V is subjected initially to reaction with a trimethylsilating agent such as MSTFA in an inert solvent such as acetonitrile under anhydrous conditions at room temperature for a period of about 10 minutes to two hours, preferably about 30 minutes.

Iodotrimetylsilane is then added and allowed to react for from 15 minutes to three hours, preferably about 30 minutes, at room temperature. A small amount of THF is then added, to decompose any residual iodotrimethylsilane. The quaternization step is then carried out by adding a protected form of the piperazine derivative of IV which is obtained from a compound of formula IV by treatment with a

- 26 - A 60 4 g trimethylsilating agent such as MSTFA in a suitable solvent such as acetonitrile. The quaternization reaction is allowed to proceed for about 30 minutes to 24 hours, preferably about two to four hours, and most preferably at room temperature. Then, an acylating agent consisting of an activated form of a carboxylic acid, for example, a thio ester such as eos

N | ,

HN—( | ~1J

S oO is added. The reaction mixture is then stirred for a period of from 2 to 24 hours. The mixture is then concentrated to dryness under reduced pressure, and the residue is redissolved in acetonitrile. Addition of methanol then solvolyzes the trimethylsilyl protecting groups and . precipitates the quaternary iodide of formula IA. In the case where other protecting yroups are present, for example, in the substituent R,, those protecting groups are then ’ ‘removed by methods known in the art. For example, if a tert.-butyl ester is present, it is cleaved by treatment with trifluoroacetic acid-anisole. Finally, as in Scheme I, the product of formula IB is obtained after aqueous reaction : with a base such as sodium bicarbonate or sodium phosphate buffer. as

AO PL

- 27 -

Scheme 111

R, R

RINH GS IV + MSTFA RINH GZ _S ~~

I Ce I

CO;R CO,R CH, I° vil lA 1) Deprotection RyNH fe Ss 9 if necessary pd J N

A — Er — i 0 N N 2) Aqueous base or butler CO; CH,

IB

Scheme 111 :

Compounds of formula VII which are starting materials for this scheme are prepared by the procedures described in

U.S. Patent No. 4,266,049 (R. Bonjouklian, May 5, 1981). In this sequence. the piperazine derivative of formula IV is converted to a protected form by reaction with a trimethyl- silating agent such as MSTFA in a suitable solvent, such as acetonitrile, under anhydrous conditions. Alternatively,

CL 2664s conventional protecting groups which are readily removable under mild conditions can be used to protect reactive functionalities in the piperazine derivative of formula IV.

For example, a carboxylic acid can be protected as a tert.butyl or p-nitrobenzyl ester. The reaction of VII with the protected form of IV in an inert solvent such as acetonitrile provides the quaternary iodide IA. Further treatment as necessary removes the protecting groups; for example, when R is tert.butyl, the ester is cleaved with trifluoroacetic acid-anisole. Subsequent treatment with aqueous sodium bicarbonate or sodium phosphate buffer gives a compound of formula I, in which other carboxylic acid functions in R, or Q may be converted to salts, depending . upon the pH.

Compounds of formula I in which m is 1l(sulfoxides) or 2(sulfones) are prepared from compounds of formula 1 in which m is 0, using oxidation procedures known to those skilled in the art, for example, oxidation with op meta-chloroperoxybenzoic acid.

The invention is further illustrated in the following examples, which are not intended to be limiting.

29 - 260d 5

EXAMPLE 1

Preparation of [6R-trans]-4-[3-cacrboxy-1-(2-fluorocethyl)- 6,8-difluoro-1,4-dihydro-4-oxoquinolin-7-yll-1-[[2-carboxy-

B8-oxo-7-[(phenoxyacetyl)amino]-5-thia-1-azabicyclo-[4.2.0]}oct- 2-en-3-yl]lmethyl}l-l-methylpiperazinium iodide 0

F COyH

H H

CL NH T 7 s

A fy or + 0 NF NA F \ : 0

CO,H CHy - F

Under an argon atmosphere, a mixture of 406 mg (1 mmol) of [B6R-trans]-3-(acetyloxy)methyl]-8-oxo-7-[(phenoxy- acetyl)amino]-5-thia-1l-azabicyclo[4.2.0])-oct-2-en-2-cacboxy- lic acid, 2 ml of dry methylene chloride, and 0.60 ml (3 mmol) of N-methyl-N-(trimethylsilyl)trifluoroacetamide ‘(MSTFA) was stirred for one hour; 0.28 ml (2 mmol) of 1iodotrimethylsilane was then added, and the mixture stirred for 2 hours. The solution was then concentrated to dryness under reduced pressure, and the residual oil dissolved in 2 ml of acetonitrile. Five drops of anhydrous THF were added, and the mixture was stirred for 5-10 minutes. A solution prepared from 111 mg (0.3 mmol) of 6,8-difluoro-1-(2- fluoroethyl)-1,4-dihydro-7-(4-methyl-l-piperazinyl)-4-oxo-3- quinolinecarboxylic acid, 0.11 ml (0.6 mmol) of MSTFA and 1 ml of acetonitrile was added, and stirring was continued for 2 hours. The mixture was chilled in ice, and approximately 100 mg of methanol were added. The solid which precipitated was filtered, washed with acetonitrile, and dried under reduced pressure to obtain the title compound: NMR

2601€ : - 30 - (Me, S0-d.) § 3.15 (s, 3H, NCH, ), 3.45-3.85 (m, 9H, 4 Xx NCH, and CH of SCH,). 3.95 (d, 1H, J gem = 16.5 Hz,

CH of SCH,). 4.39 and 4.77 (AB, 2H, J gem = 13Hz, NCH,). 4.61 and 4.64 (AB, 2H, J gem = 15 Hz, OCH, CO). 4.83-5.07 (m, 4H, NCH, CHF), 5.24 (da, 1H, J = 5Hz, CH), 5.82 (44, 1H, J = 5 and 7 Hz, CH), 6.95 (d, 2H, J = 8 Hz, Ar), 6.97 (t, 1H, J = 8 Hz, Ar), 7.29 (t, 2H, J = 8 Hz), 7.96 (d, 1 H,

J = 12 Hz, Ar). 8.91 (s, lH, =:=CH-), 9.18 (d, lH, J = 7 Hz,

NH); IR (KBr) 3400, 1788, 1728, 1700, 1612, em: mass spectrum m/z 716 (cation).

EXAMPLE 2 . . Preparation of [6R-[6a,7B(Z2)11-1-[[7-{[[(2-amino-

A4-thiazolyl)[l-(l-carboxy-l-methyl)ethoxylliminolacetyl]amino] 2-carboxy-8-oxo-5-thia-l-azabicyclof4.2.0]oct-2-en-3-y1]- methyl]-4-[3-carboxy-1-(2-fluorcethyl)-6,8-difluoro-1,4- dihydro-4-oxoquinolin-7-yl]l-l-methylpiperazinium hydroxide inner salt monosodium salt

Xo ;

NG CO, Na F CO,H nt . : N NH HEN oN N

CN YY FLA

Ss 0 = F 0 N co," CH, F ‘ Under an argon atmosphere, a mixture of 5.12 g (8 mmol) of [6R-[6a,7B(Z)]]-3-[(acetyloxy)methyl]-7-[([(2-amino-

A-thiazolyl)[l-(l-carboxy-l-methyl)ethoxyliminoJ]acetyl}ami- no)-8-o0xo-5-thia-l-azabicyclo[4.2.0)oct-2-ene-2-carboxylic acid trifluoroacetic acid salt, 48 ml of dry acetonitrile, and 12 ml (64 mmol) of MSTFA was stirred for 30 minutes; 2.0 ml (14 mmol) of iodotrimethylsilane was added dropwise, and the mixture was stirred for 30 minutes. With momentary , cooling in ice, 1.12 ml (14 mmol) of dry THF was added.

266 ts - 31 -

After 10 minutes, a solution prepared from 2.27 g (6 mmol) of 6,8-difluoro-1-(2-fluoroethyl)-1,4-dihydro-7- (4-methyl-l-piperazinyl)-4-oxo-3-quinolinecarboxylic acid, 24 ml of acetonitrile, and 1.28 ml (7.2 mmol) of MSTFA was added, and the mixture stirred for 1.5 hours. The mixture was concentrated under reduced pressure, and the residual oil was dissolved in 40 ml of acetonitrile. With ice-cooling, 4 ml of methanol were added, resulting in a thick precipitate. After settling for a few minutes, the precipitate was filtered, and washed with four 10 ml portions of acetonitrile. After drying, the solid was triturated with 60 ml of methanol, filtered, and washed with four 10 ml portions of methanol. The solid (5.7 g) thus , . obtained, was suspended in water, and aqueous sodium bicarbonate was added to bring the pH to 7. The solution of crude product was purified by Cis reverse phase HPLC in three steps. First a column of 50 g of Waters C,g-Silica was used, with water followed by 30% acetonitrile in water as eluant. Then, using a 0.025 molar pH 7 buffer-aceto- og nitrile gradient, the product was further purified by HPLC on a Waters Prep SO00A with Cig columns. Finally, the product was desalted on a flash column of 60 g of Cis silica with water and 20% acetonitrile in water as eluants.

After concentrating under reduced pressure to eliminate the organic solvent, and freeze-drying, 1.0 g of the title compound was obtained: NMR (Me, S0-d6-D,0) $ 1.37 (s, 3H, CH,). 1.44 (s, 3H, CH,). 3.10 (s. 3H, NCH, ). 3.39 and 3.88 (AB, 2H, J gem = 16.5 Hz, SCH, ). 3.40-3.70 (m, 8H, 4 x NCH, ). 4.12 and 5.17 (AB, 2H, J gem = 12.5 Hz,

NCH.) 4.62-4.94 (m, 4H, NCH, CHF), 5.15 (4d, 1H, J =

SHz, CH), 5.73 (d, lH, J = 5Hz, CH) 6.74 (s, 1H, Ar), 7.83 (d, 1H, J = 12 Hz, Ar), 8.47 (brs, lH, = CH-):; IR (KBr) 3400, 1772. 1618, 1595 en”; mass spectrum m/z = 859 (M + mt.

a2 > CC Ay

EXAMPLE 3

Preparation of [6R-[6a,78(Z))]1-1-([{7-[[(2-amino-4- thiazolyl)(methoxyimino)acetyllaminol-2-carboxy-8-0x0-5-thia- l-azabicyclof4.2.0]-oct-2-en-3-yl]lmethyl]-4-(3-carboxy-l-ethyl -6-fluoro-1,4-dihydro-4-ox0-7-quinolinyl)-l-methylpiperazinium iodide } : 0)

NOCH, F CO,H

H H

HaN— TY F A

S © NF NS § : 0 co,H CHa -

Under an argon atmosphere, a mixture of 273 mg (0.6 mmol) of [6R-[6a,7B(Z)])])-3-[(acetyloxy)methyl]-7-[[(2- amino-4-thiazolyl)(methoxyimino)acetyl]amino}-8-oxo-5-thia- l-azabicyclo[4.2.0)oct-2-ene-2-carboxylic acid, 2 ml of dry ne ‘methylene chloride, and 0.45 ml (2.4 mmol) of MSTFA was o5 stirred for 2 hours; 0.17 ml (1.2 mmol) of iodotrimethyl- silane was added, and stirring was continued for another 2 hours. The mixture was concentrated under reduced pressure, and the residual oil was dissolved in 2 ml of dry aceto- nitrile. A few drops of dry THF were added, and the mixture stirred for 15 minutes. A solution prepared from 60 mg (0.18 mmol) of l-ethyl-6-fluoro-1,4-dihydro-7-(4-methyl- -1l-piperazinyl)-4-oxo-3-quinolinecarboxylic acid, 1 ml of dry acetonitrile, and 0.07 ml (0.36 mmol) of MSTFA was added, and stirring was continued for 2 hours. Dropwise addition of 125 mg of methanol caused the product to precipitate. The solid was filtered, washed with four 1l-ml portions of acetonitrile, and dried under reduced pressure.

Cas 266 HE

After trituration with methanol and drying under reduced pressure, the title compound was obtained: NMR (Me ,50-d6) 6 1.45 (brt, 3H, CH, of NEt), 3.15 (s, 3H, NCH, ). 3.50-3.95 (m, 9H, 4 x NCH, and CH of SCH, ). 3.85 (s, 3H, . 5 OCH,). 3.99 (d. 1H, J gem = 16.5 Hz, CH of SCH, ). 4.43 and 4.70 (AB, 2H, J gem = 14 Hz, NCH), 4.61 (brq, 2H,

CH, of NEt), 5.31 (4, 1H, J = 5 Hz, CH), 5.94 (dd, lH. J = and 8 Hz, CH), 6.76 (s. lH, Ar), 7.30 (br, 2H, NH), 7.31 (d4, 1H, J = 6.5 Hz, Ar), 8.03 (d, 1H, J = 12.5 Hz, Ar), 9.03 (s, lH, = CH-). 9.68 (d, lH, J = 8 Hz, NH): IR (KBr) 1785, 1720, 1680, 1628, em”; mass spectrum m/z = 729 (cation). . EXAMPLE 4

Preparation of [6R-[6a,78(Z)]1-1-[7-[[[(2-amino-4- thiazolyl) (methoxyimino)acetyllamino]-2-carboxy-8-0x0-5-thia- l-azabicyclo[4.2.0]oct-2-en-3-yl]lmethyl]-4-[{3-carboxy-6,8-di- fluoro-1-(2-fluorcethyl)-1,4-dihydro-4-oxo-7-quinolinyl]-1- methylpiperazinium iodide 0

NOCH, F CO,H

CN NHI Ig ~~ N . Han © A - N 0

CO,H CHa - F

A mixture of 84.6 mg (0.186 mmol) of [6R-[6a,7B(Z)]]- 3-[(acetyloxy)methyl]-7-[[(2-amino-4-thiazolyl)(methoxyimino)- acetyl)amino)-8-o0x0-5-thia-1l-azabicyclo[4.2.0]oct-2-ene-2-car- boxylic acid, 4 ml of dry methylene chloride, and 0.148 ml (0.8 mmol) of MSTFA was stirred for one hour. To the resulting solution was added 0.0625 ml (0.46 mmol) of

, &

Ca ALG © jodotrimethylsilane. The mixture was stirred for 2 hours, and concentrated to dryness under reduced pressure. The residue was dissolved in 2 ml of dry acetonitrile, and a few drops of dry THF were added. The mixture was stirred for 3 minutes, and then a solution prepared from 36.9 mg (0.10 mmol) of 6,8--difluro-1-(2-fluoroethyl)-1,4-dihydro-7-(4- methyl-l-piperazinyl)-4-oxo-3-quinoline carboxylic acid, 0.040 ml (0.215 mmol) of MSTFA, and 2.0 ml of dry acetonitrile was added. The mixture was stirred for 3 hours. After addition of 0.50 ml of methanol, a precipitate formed. The solid was filtered, washed repeatedly with acetonitrile, and dried to obtain 87.5 mg of the title compound: NMR (Me, 50-d6) 6 3.15 (s, 3H, NCH, ). . 3.50-3.90 (m, 9H, 4 x NCH, and CH of SCH,). 3.86 (s, 3H,

OCH, ). 3.96 (4, lH, J gem = 17 Hz, CH of SCH,). 4.42 and 4.73 (AB, 2H, J gem = 14 Hz, NCH). 4.85-5.10 (m, 4H,

NCH, CHF). 5.29 (d, 1H, J = 5 Hz, CH), 5.93 (dd, 1H, J = 5 and 7 Hz, CH), 6.86 (s, lH, Ar), 7.24 (s, 2H, NH, ). 7.98 (d, 1H, J = 12 Hz, Ar), 8.94 (s, lH = CH-), 9.67 (d, 1H, J = op 7Hz, NH); 1R (KBr) 3420, 1775, 1720. 1675, 1618 en” l. : EXAMPLE 5 " Preparation of 6R-[6a,7R(Z)-1-[7-[[[(2-amino-4- thiazolyl) (methoxyimino)acetyllamino}l-2-carboxy-8-oxo- 5-thia-l-azabicyclof4.2.0]oct-2-en-3-yljmethyll]l-4-[3-car- boxy-6,8-difluoro-1-(2-fluoroethyl)-1,4-dihydro-4-0ox0-7-qui- nolinyl]-l-methylpiperazinium monosodium salt

Oo

NOCH, F CO,” Na*

H H

N HE Ts wl TY F AT 1 s o N N F N 0

Co,- Cha F

Cas 20G&E - NA suspension of 900 mg of the compound prepared in

Example 4 in water was neutralized with 0.1 N sodium hydroxide, and the resulting solution was freeze-dried. The residue was purified by Cin teverce phase HPLC on a Waters

Prep 500A, eluting with a water-acetonitrile gradient

Co (0-40%). Evaporation and lyophilization of the appropriate fractions afforded 344 mg of the title compound: NMR (Me, 50-d6) $ 3.10 (s, 3H, NCH.) . 3.40-3.76 (m, 9H, 4 x

NCI, and Cli of sci). 3.84 (d, 1H, J = 16 Hz, CH of

SCIL, ) . 3.04 (ss, 3, oct), 4.10 and 5.19 (AB, 2tl, J gem = 14Hz), 4.70-4.94 (m, 411, NCH, Cl F), 5.13 (d, 1H, J = 5tiz, Cll), 5.65 (dd, 111, J = 5 and 7Hz, CH), 6.74 (8s, 1H.

Ar), 7.23 (s, 211, NH, ) 7.82 (d, 1H, J = 12Hz, Ar), 0.55 . (s. ll, -Ccnl-), 9.50 (d, 1H, J = 7Hz, NH): IR (KBr) 34lo0, “ 1772, 1665, 1618 em” Ll: mass spectrum m/z 787 (M + in”.

EXAMPLE 6 . Preparation of (6R-trans)-4-{3-carboxy-6,8-difluoro-1-(2- : 20 fluoroethyl)-1,4-dihydro-4-oxo-7-quinolinylki=[(13-(1,1~dimethyl- ethoxy) carboriy1-7-(formylamino) -8-oxo-S-thia-1-azabicvclo(4.2.0]- oct-2-en-3-yllmethyl]-1-methylpiperazinium iodide or : ‘ 0, 0 F CO,H oun 5

Lens oxy

Ji NS F \ 40 © Le, I- F

A

A mixture of 0.87 g (2.35 mmol) of 6,8-difluoro-1-(2- fluoroethyl). 1,4-dihydro-7-(4-methyl-l-piperazinyl)-4-oxo-3- quinolinecarboxylic acid, 0.51 ml (2.6 mmol) of MSTFA, and S . BAD oricing

J

3-6GUE ~ 36 - ml of dry acetonitrile was stirred for 30 minutes; 1.00 g of (6R(6a,78]-7-formylamino-3-iodomethyl-B8-0ox0-5-thia-1- azabicyclo[4.2.0)oct-2-ene-2-carboxylic acid l,l1-dimethylethyl ester was added, and stirring was continued for 24 hours. The resulting precipitate was filtered, and discarded. The mother liquor was absorbed onto a column of 5 g of C,g-slilica. ‘After elution with water, 10%- and 20%-aqueous methanol, the appropriate fractions were combined and concentrated under reduced pressure to yield a precipitate. After filtration and drying, 380 mg of the title compound was obtained: IR (KBr) 3440, 1785, 1720, 1610 cmb; mass spectrum m/z = 666 (cation).

EXAMPLE 7

Preparation of (6R-trans)-4-[3-carboxy-6,8-difluoro- 1-(2-fluoroethyl)-1,4-dihydro-4-oxo-7-quinolinyl]-1-[[2- carboxy-7— (formylamino)~-8-oxo-5~-thia-l-azabicyclo[4.2.0] oct-2-en~-3-yl Jmethyl ]-1l-methylpiperazinium trifluoro- "acetate salt . 0 * 0 F CoH . II H H 5 HCNH tS : ON N +

N__~ NA F 0

CO,H CHj F

CF,CO,”

NA mixture of 200 mg of the compound from lixample 6, 0.2 ml anisole, and 2.5 ml of trifluoroacetic acid was stirred for 3 hours at room temperature. After filtration of the insoluble portion. the solution was concentrated under reduced pressure. The residue was dissolved in 10 ml of

LAD ORIGINAL

Ee as

- 37 - AGC AF acetonitrile, and 200 ml of ether was added to precipitate the product. After filtration and drying, 135 mg of the title compound was obtained: NMR (Me ,S0-d6) 5 3.14 (s, 3H, NCH,). 3.50-3.85 (m, 9H, 4 x NCH, and CH of SCH,). 3.95 (d., 1H, J gem = 16.5 Hz, CH of SCH, ). 4.35 (4d, 1H, J gem = 13Hz, CH of NCH). 4.82-5.05 (m, 5H, NCH, CH, F and CH of NCH). 5.23 (d, 1H, J = SHz, CH), 5.84 (dd, 1H,

J = 5 and 7Hz, CH), 7.96 (d, 1H, J = 13Hz, Ar), B.l17 (s., lH,

NCHO). 8.92 (s, 1H, =CH-), 9.11 (d, 1H, J = 7Hz, NH): IR 3400, 1780, 1720, 1685 em 1; mass spectrum m/z = 610 (cation).

Example 8

Preparation of [6R-{6a,78(Z)]]1-1-[[7-[[(2-amino-4- thiazolyl)[fl,1-dimethyl-2-(1,1-dimethylethoxy)-2-oxoethoxy] iminoJlacetyljamino]-2-carboxy-8-0xo-5-thia-1-azabicyclo[4.2.0] oct-2-en-3-yllmethyl]-4-[3-carboxy-(2~-fluorpethyl)-6,8- difluoro-1,4-dihydro-4-oxoquinolin-7-ylj-l-methylpiperazinium lodide

Hoot g ng CO F CO,H

TRE

+ PS FN N w= JT A

Ss 0 NA N F N 0 co,H CHa \- F

Under an argon atmosphere, a mixture of 272 mg (1 mmol) of 7-aminocephalosporanic acid, 0.67 ml (3.6 mmol) of MSTFA and 3 ml of dry acetonitrile was stirred for 30 minutes: 0.25 ml (1.75 mmol) of iodotrimethylsilane was then added and stirring was continued for another 30 minutes. The mixture was cooled momentarily, and 0.14 ml (1.75 mmol) of ~~ BAD ORiginy,

Cag 206042 anhydrous THF was added. After 10 minutes, a solution prepared from 277 mg (0.75 mmol) of 6,B-difluoro-1-(2- fluoroethyl)-1,4-dihydro-7-(4-methyl-1-piperazinyl)-4-oxo0-3- quinolinecarboxylic acid, 0.17 ml (0.9 mmol) of MSTFA and 3 ml of dry acetonitrile was added. Stirring at room temperature was continued for 2.5% hours; 478 mg (1 mmol) of 2-{[[1-(2-amino-4-thiazolyl)-2-[(2-benzothiazolyl)thio]-2- oxoethylidenejamino]oxy]lmethylpropanoic acid 1,1-dimethylethyl ester and 4 ml of dry acetonitrile were : 10 added, and the mixture was stirred overnight. After filtration to remove a small amount of insoluble solid, the mixture was concentrated to dryness under reduced pressure.

The residual oil was redissolved in 4 ml of acetonitrile, . and with ice cooling, 0.16 ml of methanol was added. After stirring for one minute and standing for 3 minutes, the precipitated solid was filtered. After washing with three 3-ml portions of acetonitrile, and drying under reduced pressure, 530 mg of the title compound was obtained: NMR (Me, 50-d6). § 1.36 (s. 12H, t-Bu and CH;). 1.40 (s. o0 3H, CH,). 3.12 (s., 3H, NCH). 3.40-3.86 (m, 9H, 4 x

NCH, and CH of SCH, ). 3.96 (d, 1H, J gem = 1l6Hz, CH of

SCH). 4.40-4.66 (AB, 2H, J gem = 13Hz, NCH, ). 4.62-5.06 (m, 41H, NCH, CHF). 5.26 (d, 1H, J = 5Hz, CH), 5.93 (44, 14, J = 5 and 7Hz, CH), 6.68 (s, 1H, Ar), 7.25 (s., 2H,

NH, ), 7.92 (d, 1H, J = 12Hz, Ar), 8.88 (s, lH, =CH-), 9.44 (d, 1H, J = 7Hz, NH). rd ‘ ‘

Cae AEG

Example 9

Alternate Synthesis of [6R-[6a,78(Z)]1]1-1-{{7-[[[(2- amino-4-thiazolyl)[l-(l-carboxy-l-methyl)ethoxyliminolacetyl]- amino]-2-carboxy-8-oxo-5-thia-l-azabicyclof4.2.0]oct-2-en-3- yl]lmethyl]l-4-[3-carboxy-1-(2-fluoroethyl)-6,8-difluoro-1,4- dihydro-4-oxoquinolin-7-yl]j-l-methylpiperazinium hydroxide inner salt monosodium salt © 5 C0" Net F I CO,H wnt ¢ | :

N PS wld TY F AA s o N NA F N o

A solution of 102 mg of the compound prepared in Example 8 in 0.4 ml of anisole, 1.5 ml of methylene chloride, and o9 1.5 ml of trifluoroacetic acid was kept overnight at 0°C.

After filtering, concentrating to dryness under reduced pressure, adding methylene chloride and again concentrating to dryness, the residue was triturated with ether to obtain a solid. The solid was dissolved in sodium phosphate buffer of pH 7, and purified by reverse phase HPLC, to obtain the title compound having an NMR spectrum similar to that of the product obtained by the previously described route (Example 2).

Claims (1)

1. CLAIMS: “ LA compound of the formula I Ry I LN PN ~~ COOH yw [ I | .

   - . ¥ . 8 TN SN - ! to , 5) . od a - Bye td 1 [ _y] i ~~ Rig Igy “Seu, : 0 1 ony

   2 . CoO hn wherein Ry 1s furmnyl, phenoxyacetyl or a group of the formula 6 0 il “fn C amram | meme Name O evens Plyag Ryo wherein Rey is a substituted 3, 6- or 7-membered heterocyclic ring ‘containing 1, 2, 3 or 4 hetero atoms selected from the group consisting of nitrogen and sulfur wherein the heterocyclie ring, is substituted by amino, and Ryo is lower alkyl or corboxy-lower alkyl, Rag represents hydrogen or halogen; Ray represents lower alkyl or halo lower alkyl; Ray represents halogen; A is a pharmaceutically acceptable anion, and pharmaceutically acceptable salts and hydrates thereof. TOA compuad as an elaim 1, wherein Rag is hydrogen, bromiuve, : chlorine or ftuoiine. Ry is lower aflkyl or hate-lower alkyl, and R33 is chlorine or fluorine. : Joa compound as in claim 2. wherein R306 hydrogen or fluorine, Ray is a5 ethyl or fluotoethyl and Ras is flusrine. LAD ORIGINAL canna

   4. A compound as in claim 1 wherein R3is the formyl group.

   3. A compound ag in claim 1 wherein Ry is the phenoxyacotyl proup.

   6. A compnnd as in claim 1 wherein Rygy is of the formula = : A - H,N S wherein Rg is lower alkyl or a group of the tormula ie NY op COO fa wherein Rpg and Ry3 are lower alkyl.

   7. A compound as in claim 1 of the formula [6R-trans]-4-[3-carboxy- 1-(2- Muoroethy)-6,8-diffuoro IL4-diliy dro-d-oxoquinolin-7-vi [| [2-carboxy-8- ux0o-7-1(phenoxyacety amino )-5-thia- t-azabicyclo{4.2.0Joct-2 en-3-y1] methyll-Tamethylpiperazinium iodide.

   8. A compound as in claim 1 of the formula (6R-160,7MZ)-1-{7-11(2- amino-4-thiazoly DH -(1-carboxy-1-methyDethoxy liminolacety! amino )-2- ! carboxy-8-oxo-5-thia-1-azabicyclol4.2.0loct-2-en-3-yl methyl |-4-[3- carboxy -1- -Afluoroethyl)-6,8-diffuoro-1,4 -dihydro-4-oxogquinolin-7-yl}- I-methyl- piperazinium hydropide inner salt monosodium salt.

   9. A compound an iu claim 1 of the tormula (6R-[6u,78(Z)J)~|-([7-{((Z-amino 4-thinzo Ly l) (methoxy iminv)acety! amino] 2-cacboxy g-oxv-5-thia-1- azabicyclo-

   [4.2.0)Joct-2-en-3-ylimethyl] a. (3 -carboky l-athyl 6-f1uo- rol, 1-dihydro-4-oxo-7 quinnlj nyl) J-methylptporazinium lodiae. : - / rt sl : Co Cry NRININAL {

   AC p -42-

   10. A compound as in claim 1 of the formula (6R-[6u, /R(LY))-L={T7-L{[(2~aminv-4-thiazolyl) (moLthoxy- imino)acetyl Jawino |-2-carboxy -f-oxo-5-thia-l-azabicyclo-

   (4.2.0]oct-2 -en-3-yllmothyl)-4-(3- carboxy-6,8.-difluoro-\- (2-fluoroerhyl)-1,4-dihydro--4.-0xo~7-quinolinyl j-l-mothyl- piporcaziniuwe lodide.

   11. A compound au in claim 1 of the formula (OR-[6a,7B(L)])-1-[7-1Ll(2~amino-4-thiazolyl) (methoxy- imino)acetyl)awino]-2-carboxy-#-oxo-5-thla-l-azabicyclo

   (4.2.0)Joct-2-en-3.ylimethyl)-4 [3-~carboxy-6,8-ditfluoro 1- (2-fluorvathyl)-1,4-dihydro.-4-oxo~7-quinolinylj}-l-wethyl- piperazinlum monosodium galt. oo .

   12. A compound as in claim 1 of the formula (6R-trans)-4-(3-carboxy-6,8~-difluoro-1-(2-£luoroethyl)-

   l.4-dihvdro- 1-oxo-7-quinotinyl]l-1-([2-().l-dimothyl- ethoxy)carbonyl-7.-(formylamino)-48-oxo-5-thia-l-azablcyclo-

   [4.2.0)oct-2-en-3 -yllmothyl}-l-methylpipecrazinium lodide.

   13. A compound as in claim 1 of the formula (6R-trans)--4-[3-carboxy-6,8-difluoro-1-(2- °~ fluoroethyl)-1.4-dihydro-d4-oxo~7-quinolinyl}-?-({2- carboxy~7- (formylamino)-8-oxo-6-thia-l-azabicyclu(d4.2.0] oct -2-on-3-ylJmethyl]-l-methylpiperazinium trifluorcaceatate all,

   14. MN compound as ih claim 1 of the formula (6R- [ 6a, 7B(ZY1)-1-[[7- {ll (Zz~amino-4-vhlazolyl [(1,} dimethyl-2-(L,l-dimethylethoxy)-2-oxoathoxy]imino)- 1] acetyllamino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct- 2-en-3-yl]-methyl]-4-[3-carboxy(2-fluoroethyl)-6,8-difluorc- 1,4-dihydro-4-oxoquinoline-7-yl]-l-methylpiperazinium iodide. HARRY ALLEN ALBRECHT DENNIS DALTCN KEITH , CHUNG-CHEN WEI MANFRED WEIGELRE ROXANA YANG (Inventors) BAD ORIGINAL