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WO1996038450A1 - Cephalosporin antibiotics - Google Patents

Cephalosporin antibiotics Download PDF

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Publication number
WO1996038450A1
WO1996038450A1 PCT/US1996/008928 US9608928W WO9638450A1 WO 1996038450 A1 WO1996038450 A1 WO 1996038450A1 US 9608928 W US9608928 W US 9608928W WO 9638450 A1 WO9638450 A1 WO 9638450A1
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WO
WIPO (PCT)
Prior art keywords
amino
group
alkyl
compound
cephem
Prior art date
Application number
PCT/US1996/008928
Other languages
French (fr)
Inventor
Burton Christensen
In-Seop Cho
Tomasz Glinka
Scott Hecker
Ving J. Lee
Zhijia J. Zhang
Original Assignee
Microcide Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/455,969 external-priority patent/US5698547A/en
Priority claimed from US08/457,673 external-priority patent/US5688786A/en
Application filed by Microcide Pharmaceuticals, Inc. filed Critical Microcide Pharmaceuticals, Inc.
Priority to AU59855/96A priority Critical patent/AU5985596A/en
Publication of WO1996038450A1 publication Critical patent/WO1996038450A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D477/00—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D477/00—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring
    • C07D477/10—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2
    • C07D477/12—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2 with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached in position 6
    • C07D477/16—Heterocyclic compounds containing 1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbapenicillins, thienamycins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulphur-containing hetero ring with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 4, and with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2 with hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached in position 6 with hetero atoms or carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 3
    • C07D477/20—Sulfur atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D499/00—Heterocyclic compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. penicillins, penems; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D499/88—Compounds with a double bond between positions 2 and 3 and a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D503/00—Heterocyclic compounds containing 4-oxa-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. oxapenicillins, clavulanic acid derivatives; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D505/00—Heterocyclic compounds containing 5-oxa-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. oxacephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • -NR 4 R 5 ' and Z is selected from the group consisting of -CH 2 (X) m -, -C(NOR 6 )-, -CH(OR 7 )-, -C(CHC0 2 R 8 )- and -CH(NR 9 R 10 ) - ;
  • n is selected from the group consisting of 0 and 1;
  • R 2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl;
  • R 11 is selected from the group consisting of H, halogen, alkyl, hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroaryliumalkyl;
  • R 17 is H or alkyl.
  • Preferred compounds include those compounds wherein R 11
  • the present invention includes methods for treating a bacterial infection in a mammal arising from bacteria resistant to ⁇ -lactam antibiotics comprising administering to a mammal suffering from such an infection a therapeutically effective amount of a compound of Structure II.
  • the present invention provides methods for using a thiolate nucleophile of the structure
  • heterocycle denotes a chain of carbon and at least one non-carbon atoms which together form one or more aromatic or non-aromatic rings having preferrably between about 6-14 atoms, such as, e.g. , furyl, thienyl, imidazolyl, indolyl, pyridinyl, thiadiazolyl, thiazolyl, piperazinyl, dibenzfuranyl, dibenzthienyl . These rings may be optionally substituted with one or more functional groups which are attached commonly to such rings, such as, e.g..
  • heterotricycle denotes an aromatic heterocyclic substituent as defined above which comprises three aromatic rings.
  • heterocyclecarbonyl denotes the group
  • ⁇ -lactam resistant bacteria refers to bacteria against which a ⁇ -lactam antibiotic has an minimum inhibitory concentration (MIC) greater than 32 mg/ml.
  • R 6 is selected from the group consisting of hydrogen, methyl, 2-fluoroethyl, cyclopropylmethyl, allyl, dichloroallyl and cyclopentyl
  • R 3 is selected from the group consisting of phenyl, 2-thienyl, 2-furyl, 2-aminothiazol-4-yl, and 2-aminothiadiazol-4-yl;
  • R 13 - R 17 are H or lower alkyl .
  • R 13 -R 17 are hydrogen.
  • a suitable effective dose of the compound of the invention will be in the range of 0.1 to 1000 milligram (mg) per recipient per day, preferably in the range of 1 to 100 mg per day.
  • the desired dosage is preferably presented in one, two, three, four or more subdoses administered at appropriate intervals throughout the day. These subdoses can be administered as unit dosage forms, for example, containing 5 to 1000 mg, preferably 10 to 100 mg of active ingredient per unit dosage form.
  • the compounds of the invention will be administered in amounts of between about 2.0 mg/kg to 250 mg/kg of patient body weight, between about one to four times per day.
  • Disposable sterile l ⁇ L loops were used to inoculate test plates, with each isolate in a designated grid on the agar plate.
  • An alternate method of inoculation involved the use of a replica plater, a device with 48 steel pins allowing the simultaneous inoculation of multiple isolates. After the spots had dried, the plates were incubated at 35-36°C for 16-20 hours. Endpoints were assessed as the minimum inhibitory concentration (MIC) of antimicrobial agent.
  • MIC minimum inhibitory concentration
  • aureus 76 (Meth R )(lac + ) >32 4 4 4 8 2 8
  • mice Groups of 5 female Swiss-Webster mice (Simonsen, Gilroy, CA) each are challenged by the intraperitoneal (IP) route with tenfold increments of a bacterial inoculum. This permits calculation of the mean lethal dose (LD 50 ) and the LD 100 .
  • mice are challenged IP with an LD 100 titer of bacteria.
  • groups of 10 mice each are treated subcutaneously with two-fold increments of the test drug and an antibiotic of known efficacy in mice and humans (i.e., positive control) . Mice are observed for 72h. Those alive at 72h are considered long term survivors.
  • the total drug dose in mg/kg that protects 50% of mice in a group from death is termed the mean protective dose (PD 50 ) .
  • PD 50 s are similarly determined for several pathogens.
  • the quantitative endpoints for the new drug are then compared with those obtained with reference antibiotics.
  • Six ten-fold dilutions of inoculum suspended in 0.5 mL of sterilized 7% hog gastric mucin (Sigma) are injected IP in groups of 5 mice each. A control group of 5 mice receive mucin alone. Mice are observed for 72h. Those alive at 72h are considered long term survivors.
  • the mean lethal dose (LD 50 ) and 100% lethal dose (LD 100 ) are determined by the probit test.
  • mice are challenged IP with bacterial titers that will afford an LD 100 for the test strain.
  • SC subcutaneous route
  • Drug doses can range from 0.01 to 512 mg/kg. If the drug is poorly soluble, Tween 20 or propylene glycol will be employed to solubilize the drug. Animals are observed for 72h.
  • the 50% protective dose (PD 50 ) is calculated in mg/kg by the probit method.
  • the PD 50 is the same as the 50% effective dose (ED 50 ) and the 50% curative dose (CD 50 ) .
  • Samples of blood from the hearts of all animals that die and from half the mice that survive are cultured on brain-heart infusion agar. Animals that received a protective dosage of the test drug will be alive at 72h, although they may appear moderately ill to very ill during the observation period. Infected, placebo-treated control mice and those receiving non-effective i.e. lower dosages of the test drug will demonstrate a high rate of mortality. Most of these mice will die within 6 to 48 h. Those alive at 72h will be considered long term survivors.
  • the mixture was diluted with dichloromethane (20 mL) , and was washed with 5% aqueous sodium bicarbonate solution (20 mL) .
  • the organic layer was dried with sodium sulfate, and the solvent was removed with a rotary evaporator.
  • the residue was subjected to flash chromatography on silica gel (33% to 50% ethyl acetate/hexane) to afford 300 mg (45%) of the title compound.
  • the mixture was diluted with dichloromethane (20 mL) , and was washed with 5% aqueous sodium bicarbonate solution (20 mL) .
  • the organic layer was dried with sodium sulfate, and the solvent was removed with a rotary evaporator.
  • the residue was subjected to flash chromatography on silica gel (33% to 50% ethyl acetate/hexane) to afford 300 mg (50%) of the title compound.
  • Example 33 The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- (4-biphenylthio) -3-cephem-4-c arboxylate, benzhydryl ester is used.
  • Example 33 The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [4- (2-imidazolyl) phenylthio] - 3
  • Example 33 The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [3-bromo-4- (phenyl) phenylthio ] -3-cephem-4-carboxylate, benzhydryl ester is used.
  • Example 52 Using substantially the same procedure as in Example 51, the items of the following Examples 52 - 82 can similarly be prepared.
  • Example 51 The procedure of Example 51 is employed, but the reactants are 2- (iodophenythio) acetic acid and (71?) -7-amino-3- [2-iodophenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
  • Example 51 The procedure of Example 51 is employed, but the reactants are 2- (iodophenythio) acetic acid and (71?) -7-amino-3- (biphenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
  • Example 51 The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
  • Example 51 The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
  • Example 51 The procedure of Example 51 is employed, but the reactants are cyanoacetic acid and
  • EXAMPLE 82 (71?) -7- [(a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl)amino] -3- [dibenzofuran-3-ylthio] -3-cephem-4-carb oxylate, 4-methoxybenzyl ester
  • Example 83 Under substantially the same procedure as in Example 83, the items of the following Examples 84 - 88 are similarly prepared.
  • EXAMPLE 88 (71?) -7- [ (a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl)amino] -3- [4- (chloromethyl)phenylthio] -3-cephem-4 -carboxylate, 4-methoxybenzyl ester

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Cephalosporin Compounds (AREA)

Abstract

The present invention includes novel (7R)-7-(acylamino)-3-(arylthio)-3-cephem-4-carboxylic acids or their pharmacologically acceptable salts which exhibit antibiotic activity against a wide spectrum of organisms including organisms which are resistant to β-lactam antibiotics and are useful as antibacterial agents. The invention also relates to novel intermediates useful for making the novel compounds of the present invention and to novel methods for producing the novel compounds and intermediate compounds.

Description

CEPHA OSPORIN ANTIBIOTICS
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to cephalosporin antibiotics. More particularly, the present invention includes novel
( 7R) -7- (acylamino) -3- (arylthio) -3-cephem-4-carboxylic acids and their pharmacologically acceptable salts and prodrugs, their methods of production and use. These compounds exhibit antibiotic activity against a wide spectrum of organisms, including organisms which are resistant to β-lactam antibiotics.
10
2. Review of the Background Art
The following description of the background of the invention is provided to aid in the understanding of the inventionand is not admitted to be or describe prior
15 art to the invention.
Over the past three decades a large variety of antibiotics has become available for clinical use. One class of antibiotics which has seen remarkable growth
1. are the cephalosporins (shown generically below) , over
20 70 of which have entered clinical use for the treatment of bacterial infections in mammals since 1965. The cephalosporins exhibit their antibacterial activity by have been extremely effective in treating a wide variety of bacterial infections. Cephalosporins that are said to have antibacterial activity are described in U.S. Patent 3,992,377 and U.S. Patent 4,256,739.
Figure imgf000004_0001
I Unfortunately, the wide-spread and indiscriminant use of these antibiotics has led to a rapid increase in the number of bacterial strains which are resistant to these compounds. Most importantly, this resistance has emerged among clinically important microorganisms which threaten to limit the utility of presently available cephalosporin antibiotics. In particular, resistant strains of Salmonella , S. pneumonias, Enterobacteriacess, and Pseudomonas have emerged which threaten to undo many of the strides made in reducing mortality and morbidity from bacterial infections.
Bacterial resistance to cephalosporins follows three major pathways: a) the development of β-lactamases capable of inactivating the β-lactam ring of the cephalosporin; b) decreased cephalosporin penetration into the bacteria due to changes in bacterial cell wall composition; and c) poor binding to penicillin-binding proteins (PBPs) . The latter pathway is especially important, as the binding of β-lactams to PBPs is essential for inhibiting bacterial cell-wall biosynthesis. Certain Gram-positive bacteria, namely methicillin-resistant Staphylococcus aureus ("MRSA") and 5 enterococci are highly resistant to β-lactam antibiotics. Resistance in MRSA is due to the presence of high levels of an unusual PBP, PBP2a, which is insensitive, or binds poorly, to β-lactam antibiotics. The activity of β-lactam antibiotics against
10 PBP2a-containing organisms has been shown to correlate well with the binding affinity of the antibiotic to PBP2a.
Currently, the glycopeptides vancomycin and teicoplanin are primarily used for MRSA bacteremia. The quinolone
15 antibacterials and some carbapenems, such as imipenem, have been reported to be active against a few MRSA strains, but their use is restricted due to emerging resistant MRSA strains.
Experimental compounds which may possess
20 utility as anti-MRSA or anti-enterococcal bactericides include the glycylcyclines (see, e.g.. P.-E. Sum et. al .. J. Med. Chem.. 37, (1994)) , FK-037 (see, e.g.. H. Ohki et al., J. Antibiotics. 46:359-361 (1993)) , RP-59,500
(see, e.g , S.K. Spangler e_t a . , Antimicro. Agents
* 25 Chemother , 36:856-9 (1992)) , the everninomycin complex i (see. e.g , .E. Sanders et, al. , Antimicro. Agents
Chemother , 6: 232-8 (1974)) , the 2- (biaryl) carbapenems
(see, e.α ., U.S. Patent No. 5,025,006) ,
3- (benzothiazolylthio) cephems (see, e.g. , EP Application No. 527686) , 3- (thiazolylthio) carbacephe s (see, e.g.. R.J. Ternansky et. al.. J. Med. Chem.. 36:1971 (1993) and U.S. Patent No. 5,077,287) and arbekacin (S. Kondo, et al. J. Antibiotics 46:531 (1993) .
Recent advances in antiobiotics useful for treating infections caused by resistant bacteria are described in commonly owned and International application No. PCT/US95/03976, filed March 29, 1995 and U.S. applications Serial No. 08/457,643, filed June 1, 1995; Serial No. 08/455,969, filed May 31, 1995; Serial Nos. 08/413,713, 08/413,714, 08/415,065, 08/455,969,
08/413,712, 08/415,064, 08/415,069, all filed March 29, 1995; Serial No. 08/369,798, filed January 6, 1995; and Serial No.08/222, 262, filed April 1, 1994, all of which are incorporated herein by reference in their entirety, including any drawings.
SUMMARY OF THE INVENTION The present invention includes compounds, compositions and methods effective to treat infections in mammals arising from β-lactam antibiotic resistant bacteria.
In one aspect the present invention features a (7R) -7 (acylamino) -3- (arylthio) -3-cephem-4-carboxylic acid cephalosporin antibiotic and has antibiotic activity against an organism resistant to a beta-lactam antibiotic. Examples of particular classes of compounds having such activity are described below and data is presented showing the in vi tro and in vivo effectiveness of such compounds. Such compounds have a minimum inhibitory concentration (MIC) that is less that 50%, more preferably less than 10%, and most preferably less than 1% of the MIC of cefotaxime for a beta-lactam resistant organism, such as those listed in Table 1, preferably a beta-lactam resistant Staphylococcal or Enterococcal organism. Other preferred compounds are able to prevent or reduce mortality in mice infected with the beta-lactam resistant organism to a greater extent that vancomycin or cefotaxime.
In one aspect the invention features compounds of the structure :
Figure imgf000007_0001
II
and its pharmaceutically acceptable salts and prodrugs, wherein:
R1 is selected from the group consisting of -NHC ( 0) ZR3 ,
-NR4R5' and
Figure imgf000007_0002
Z is selected from the group consisting of -CH2(X)m-, -C(NOR6)-, -CH(OR7)-, -C(CHC02R8)- and -CH(NR9R10) - ;
X is selected from the group consisting of O and S;
m is is selected from the group consisting of 0 and 1;
R3 is selected from the group consisting of cyano, alkyl, aryl, heterocycle, heteroaralkyl and (CH2)nT,
n is 1 to 6,
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino;
R4"7 are independently selected from the group consisting of hydrogen, alkyl, aryl and acyl;
R8 is selected from the group of hydrogen, alkyl and aryl;
R9 and R10 are selected independently from the group consisting of hydrogen, alkyl, acyl, and heterocyclecarbonyl;
R2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl; R11 is selected from the group consisting of H, halogen, alkyl, hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroaryliumalkyl;
p is 0, 1, or 2 q is 0 or 1 r is 1, 2 or 3,
Figure imgf000009_0001
R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring; and
R17 is H or alkyl.
Preferred compounds include those compounds wherein R11
is H or halogen, R12 is NR13R14 ,
Figure imgf000009_0002
Figure imgf000009_0003
p i s o or 1 , q is 0 or 1 , r is 1 , 2 or 3, R13-R15 are H or lower alkyl, and R17 is H or lower alkyl. Especially preferred compounds include those compounds wherein R11 is hydrogen, R12 is
Figure imgf000010_0001
1, r is 1, 2, or 3, R13-R16 are hydrogen, and R17 is hydrogen or lower alkyl . There are preferably one or two [ (CH2)p(S)q(CH2)rR12] groups present. There are preferably three or four R11 groups present . Preferred pharmaceutically acceptable salts include (1) inorganic salts such as chloride, bromide, iodide, nitrate, phosphate or sulfate; (2) carboxylate salts such as acetate, propionate, butyrate, maleate, or fumarate; (3) alkylsulfonates such as methanesulfonate, ethanesulfonate, 2-hydroxyethylsulfonate, n-propylsulfonate or iso-propylsulfonat; and (4) hydroxycarboxylates such as lactate, malate, and citrate.
In another embodiment, the present invention provides for compositions comprising an amount of the compound of Structure II effective to treat bacterial infections in mammals arising from bacteria resistant to β-lactam antibiotics.
In still another embodiment, the present invention includes methods for treating a bacterial infection in a mammal arising from bacteria resistant to β-lactam antibiotics comprising administering to a mammal suffering from such an infection a therapeutically effective amount of a compound of Structure II.
In yet another aspect the present invention provides methods for using a thiolate nucleophile of the structure
Figure imgf000011_0001
wherein:
R11 is selected from the group consisting of H, halogen, alkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl;
p is 0, 1 or 2
Y is OH, (S)q(CH2)rOH or (S)q(CH2) rR18 q is 0 or 1 r is 1, 2, or 3 and R18 is a protected version of one of the groups R12,
rf NR15-16 » * o" namely, NR13R14 , .V M Y
Figure imgf000011_0002
, NR14 NlDR1"4 n
Figure imgf000012_0001
as a chemical reagent comprising the step of exposing said thiolate nucleophile to a cephem triflate of structure
Figure imgf000012_0002
wherein
R1 is selected from the group consisting of -NHC(0)ZR3,
Figure imgf000012_0003
Z is selected from the group consisting of -CH2(X)m-, -C(NOR6)-, -CH(OR7)-, -C(CHC02R8)- and -CH(NR9R10) -;
X is selected from the group consisting of 0 and S;
m is is selected from the group consisting of 0 and 1; and
R2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl; Where Y is OH, the invention further provides a method for conversion of this OH to a leaving group, and a method for reaction of this intermediate with a nucleophile of the form H (S) q(CH2) rR18 q is 0 or 1 r is 1, 2, or 3 and R18 is an optionally-protected version of one of the
groups R12 , namely , NR1 133Rr> 14
Figure imgf000013_0001
Figure imgf000013_0002
Where Y is (S)q(CH2)rOH, the invention further provides a method for conversion of the OH to a leavng group, and a method for reaction of this intermediate
with a nucleophile of the form
Figure imgf000013_0003
wherein
R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring. In preferred embodiments the substituents are the same as described herein for the compounds and the reactions are carried out as described in the examples described herein. Preferred conditions for performing such methods including concentrations, solvents, temperatures, etc. are also listed in the examples.
Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof and from the claims.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
I . Definitions
As used herein, the term "alkyl" denotes branched or unbranched hydrocarbon chains containing between one and six, preferably one and four, carbon atoms, such as, e.g.. methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, and 2-methylpentyl . These groups may be optionally substituted with one or more functional groups which are attached commonly to such chains, such as, e,g. , hydroxyl, bromo, fluoro, chloro, iodo, mercapto or thio, cyano, alkylthio, heterocycle, aryl, heteroaryl, carboxyl, alkoxycarbonyl, alkyl, alkenyl, nitro, amino, alkoxyl, amido, and optionally substituted isothioureido, amidino, guanidino, and the like to form alkyl groups such as trifluoromethyl, 3-hydroxyhexyl, 2-carboxypropyl, 2-fluoroethyl, carboxymethyl, 4-cyanobutyl,
2-guanidinoethyl, 3-N,N' -dimethylisothiouroniumpropyl, and the like.
The term "alkenyl" denotes an alkyl group as defined above having at least one double bond, e.g.. allyl, 3-hydroxy-2-buten-l-yl, 1-methyl-2-propen-1-yl and the like. The term "aryl" denotes a chain of carbon atoms an which form an least one aromatic ring having preferably between about 6-14 carbon atoms, such as, e.g.. phenyl, naphthyl, indenyl, and the like, and which may be substituted with one or more functional groups which are attached commonly to such chains, such as, e.g. , hydroxyl, bromo, fluoro, chloro, iodo, mercapto or thio, cyano, cyanoamido, alkylthio, heterocycle, aryl, heteroaryl, carboxyl, alkoxycarbonyl, alkyl, alkenyl, nitro, amino, alkoxyl, amido, and the like to form aryl groups such as biphenyl, iodobiphenyl, methoxybiphenyl, anthryl, bromophenyl, iodophenyl, chlorophenyl, hydroxyphenyl, methoxyphenyl, formylphenyl, acetylphenyl, trifluoromethylthiophenyl, trifluoromethoxyphenyl, alkylthiophenyl, trialkylammoniumphenyl, amidophenyl, thiazolylphenyl, oxazolylphenyl, imidazolylphenyl, imidazolylmethylphenyl, cyanophenyl, pyridylphenyl, pyrrolylphenyl, pyrazolylphenyl, triazolylphenyl, tetrazolylphenyl and the like. The term "heterocycle" denotes a chain of carbon and at least one non-carbon atoms which together form one or more aromatic or non-aromatic rings having preferrably between about 6-14 atoms, such as, e.g. , furyl, thienyl, imidazolyl, indolyl, pyridinyl, thiadiazolyl, thiazolyl, piperazinyl, dibenzfuranyl, dibenzthienyl . These rings may be optionally substituted with one or more functional groups which are attached commonly to such rings, such as, e.g.. hydroxyl, bromo, fluoro, chloro, iodo, mercapto or thio, cyano, cyanoamido, alkylthio, heterocycle, aryl, heteroaryl, carboxyl, oxo, alkoxycarbonyl, alkyl, alkenyl, nitro, amino, alkoxyl, amido, and the like to form rings such as, e.g., 2-aminothiazol-4-yl, 2, 3-dioxopiperazinyl, 4-alkylpiperazinyl, 2-iodo-3-dibenzfuranyl and 3-hydroxy-4-dibenzthienyl and the like. The term "heteroaromatic" or "heteroaryl" (HetAr) denotes an aromatic heterocycle as defined above.
The term "heterotricycle" denotes an aromatic heterocyclic substituent as defined above which comprises three aromatic rings. The term "heterocyclecarbonyl" denotes the group
-C(0)Het, where Het is heterocycle as defined above.
The term "alkoxyl" denotes the group -OR, where R is alkyl as defined above, such as methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, sec-butoxy, iso-butoxy, tert-butoxy, trifluoromethoxy, 3-hydroxyhexyloxy,
2-carboxypropyloxy, 2-fluoroethoxy, carboxymethoxy and cyanobutyloxy and the like.
The term "alkylthio" denotes the group -SR, where R is alkyl as defined above, such as methylthio, ethylthio, n-propylthio, iso-propylthio, n-butylthio, sec-butylthio, iso-butylthio, tert-butylthio, trifluoromethylthio, 3-hydroxyhexylthio, 2-carboxypropylthio, 2-fluoroethylthio, carboxymethylthio and cyanobutylthio and the like.
The term "acyl" denotes groups -C(0)R, where R is alkyl as defined above, such as formyl, acetyl, propionyl, or butyryl .
The term "aryloxy" denotes groups -OAr, where Ar is an aryl group as defined above.
The term "aralkyl" denotes groups -RAr, where R is alkyl and Ar is aryl, both as defined above.
The term "heteroaralkyl" denotes groups -RHetAr where R is alkyl and HetAr is heteroaryl as defined above.
The term "trialkylsilyl" denotes the group RR'R"Si-, where R, R1 and R" are alkyl as defined above. The term "trialkylammonium" denotes the group
[RR'R"N-]+, where R, R' and R" are alkyl as defined above.
The term "amino" denotes the group NRR' , where R and R' may independently be alkyl, aryl or acyl as defined above, or hydrogen. The term "amido" denotes the group -C(0)NRR', where R and R' may independently be alkyl, aryl or acyl as defined above, or hydrogen.
The term "cyanoamido" refers to the group -NH-CjN.
The term "β-lactam resistant bacteria" refers to bacteria against which a β-lactam antibiotic has an minimum inhibitory concentration (MIC) greater than 32 mg/ml.
The term "prodrug" refers to an agent that is converted into the parent drug in vivo. Prodrugs may be easier to administer than the parent drug in some situations. For example, the prodrug may be bioavailable by oral administration but the parent is not, or the prodrug may improve solubility to allow for intravenous administration.
II. Compounds of the Invention
The present invention provides compounds, methods and compositions effective to treat bacterial infections, and, especially, infections arising from bacteria which have developed resistance to conventional β-lactam antibiotics. More importantly, the present invention provides compounds, methods and compositions effective to treat bacterial infections arising from bacteria which have developed resistance to conventional cephalosporin antibiotics.
B. Preferred Embodiments of Structure II
Preferred compounds include those wherein:
(1) R1 is NHC(0)ZR3
Z is -CH2(X)m- X is S m is 1 R3 is (CH2)nT
(2) R1 is NHC(0)ZR3 Z is -C(NOR6) -
R6 is selected from the group consisting of hydrogen, methyl, 2-fluoroethyl, cyclopropylmethyl, allyl, dichloroallyl and cyclopentyl, and R3 is selected from the group consisting of phenyl, 2-thienyl, 2-furyl, 2-aminothiazol-4-yl, and 2-aminothiadiazol-4-yl;
(3) R11 is H or halogen
( 4 ) R12 is NR13RX
Figure imgf000018_0001
Figure imgf000019_0001
( 5 ) p is 0 or 1 q is 0 or 1 r is 1 , 2 or 3 ; and/or
( 6 ) R13 - R17 are H or lower alkyl .
Especially preferred compounds include those compounds wherein R11 is hydrogen, R12 is NR13R14 ,
Figure imgf000019_0002
p is 1, q is 1, r is 1 or 2, and R13-R17 are hydrogen. There are preferably one or two [ (CH2)p(S)q(CH2) rR12] groups present . There are preferably three or four R11 groups present .
Especially preferred compounds combining the above-described substituents include the following:
(71.) -7- [ (Phenylacetyl) amino] -3- (2-amidinomethylthiomethylphe nylthio) -3-cephem-4-carboxylate, inner salt;
( R) -7- [ (Phenylacetyl) amino] -3- (2- (2-amidinomethylthio) ethyl phenylthio) -3-cephem-4-carboxylate, inner salt;
( 7R) -7- [ (Phenylacetyl) amino] -3- (2-guanidinoethylthiomethylph enylthio) -3-cephem-4-carboxylate, inner salt;
( R) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-amidinomet hylthiomethylphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt;
(7i?) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-formamidoy lmethylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt; ( R) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-guanidinoe thylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt;
(71?) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-dimethylam inoethylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt;
(71?) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-isothioure idoethylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt;
(71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- (2-amidinomethylthiomethylphenylthio) -3-cephem-4-carboxyl ate, inner salt;
(71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- (2-formamidoylmethylthiomethylphenylthio) -3-cephem-4-carb oxylate, inner salt; ( IR) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- (2-guanidinoethylthiomethylphenylthio) -3-cephem-4-carboxy late, inner salt;
(71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- (2-dimethylaminoethylthiomethylphenylthio) -3-cephem-4-car boxylate, inner salt;
(7i?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- (2-isothioureidoethylthiomethylphenylthio) -3-cephem-4-car boxylate, inner salt;
(71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- (2-amidinomethylthiomethylphenylthio) -3-cephem-4-c arboxylate, inner salt;
(7J?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- (2-formamidoylmethylthiomethylphenylthio) -3-cephem -4-carboxylate, inner salt; (7J?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- (2-guanidinoethylthiomethylphenylthio) -3-cephem-4- carboxylate, inner salt;
(7i?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- (2-dimethylaminoethylthiomethylphenylthio) -3-cephe m-4-carboxylate, inner salt;
(7J?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- (2-isothioureidoethylthiomethylphenylthio) -3-cephe m-4-carboxylate, inner salt.
D. Synthesis of Compounds of Structure II
The compounds of the present invention may be readily prepared in accordance with the following schemes. However, it will be appreciated that other synthetic pathways for forming the compounds of the invention are available and that the following is offered merely by way of example, and not limitation. It will be further recognized that various protecting and deprotecting strategies will be employed which are standard in the art (see, e.g.. Green and Wuts) . Those of skill in the art will recognize that the selection of any particular protecting group (e.g. , a carboxy protecting group) will depend on the stability of the protected moiety with respect to subsequent reaction conditions .
Generally, the synthesis of the cephalosporins of the present invention may be achieved using well-known methods and readily available materials (see, e.g.. March; Larock, COMPREHENSIVE ORGANIC TRANSFORMATIONS (VCH Publishers, 1989) ; and G.I. Georg, THE ORGANIC CHEMISTRY OF β-LACTAMS, (VCH 1992) , each of which is incorporated herein by reference) . As shown below in Scheme 1, treatment of the cephem triflate 1 with the desired thiolate nucleophile 2, using standard methods such as those described in Farina e_t al. , J. Qrσ. Chem. 54:4962 (1989) and U.S. Patent No. 4,870,168 to Baker, et al. , (both of which are incorporated herein by reference) , provides the 3-thio derivative 3. Activation of the hydroxyl group, followed by displacement with a nucleophile of the general structure HS(CH2)rR12, affords derivative 4. Subsequent deprotection using procedures known to those skilled in the art affords the biologically active 4-carboxycephem 5.
Figure imgf000023_0001
1) Hydroxyl Activation
2) Displacement
Figure imgf000023_0002
Compound 1 is formed readily from commercially available starting materials, such as the reaction of (7R) -7- [ (phenylacetyl) amino] -3-hydroxy-3-cephem-4-carboxylic acid, p-methoxybenzyl or benzhydryl ester (Otsuka Chemical Co., Ltd., Otsuka, Japan) with triflic anhydride (Aldrich, Milwaukee, Wl) , using known procedures (see. e.g.. Farina; and U.S. Patent No. 4,870,168 to Baker, et al. ) Other 3-hydroxy-3-cephems may be formed from the ozonolysis of 3-exomethylene cephems using known procedures (see, e.g.. Farina) . Similarly, the thiolate nucleophile 2 may be formed using known procedures and commercially available starting materials.
The substituent R1 may be any of the groups described above and are either available commercially (e.g.. from Aldrich, Milwaukee, Wl) or can be formed using known techniques and starting materials (see, e.g., March; Larock) . These groups can be substituted for those present on the starting material by variety of well known techniques (see, e.g.. Barrett, J.C.S. Perkin I, 1629 (1979) or Chauvette, J. Org. Chem. 36:1259 (1971) , both of which are incorporated herein by reference) , such as by transamination of an existing substituent for the desired substituent, or hydrolytic removal of the existing substituent followed by reaction with a suitably reactive form of desired substituent, such as an acyl chloride. Again, the appropriate reagents and techniques will be apparent to those of skill in the art.
The carboxyl group R2 may be those protecting groups amenable to reductive cleavage, such as benzyl, p- or o-nitrobenzyl, 2, 2, 2-trichloroethyl, allyl, cinnamyl, benzhydryl, 2-chloroallyl and the like. Alternatively, R2 may be a protecting group amenable to acidic cleavage, such as t-butyl, t-amyl, trityl, 4-methoxytrityl, 4, 4 ' -dimethoxytrityl, trimethylsilyl, t-butyldimethylsilyl, phenacyl, β- (trimethylsilyl) ethyl, benzyl, 4- (or
2-methoxybenzyl, 2,4-dimethoxybenzyl, 3 , 4-dimethoxybenzyl, 2,4, 6-trimethoxybenzyl, methoxymethyl, benzhydryl, or 3, 3-dimethylallyl. Preferred protecting groups are p-methoxybenzyl, p-nitrobenzyl, allyl and benzhydryl. Such groups may be attached to the unprotected carboxyl group of the cephalosporin starting material using known reagents and techniques, such as those described in Green and Wuts.
III. Pharmaceutical Applications and Preparations According to this invention, a therapeutically or pharmaceutically effective amount of a cephalosporin and particularly, a compound of Structure II, is administered to a mammal suffering from a β-lactam resistant bacterial infection, especially resistant S . aureus, E. faecali , s or E. faecium, in an amount effective to at least partially relieve the infection. Especially important are infections resulting from β-lactam resistant strains having β-lactam resistance similar to strains such as S . aureus Col (MethR) (lac) , S. aureus 76 (MethR) (lac+) , E. fseciu ATCC 35667, or E. fascalis ATCC 29212.
The compositions containing the compound(s) of the invention can be administered for prophylactic and/or therapeutic treatments. In therapeutic applications, the compositions are administered to a patient already suffering from an infection, as described above, in an amount sufficient to cure or at least partially arrest the symptoms of the infection. An amount adequate to accomplish this is defined as "therapeutically 'effective amount or dose." Amounts effective for this use will depend on the severity and course of the infection, previous therapy, the patient's health status and response to the drugs, and the judgment of the treating physician. In prophylactic applications, compositions containing the compounds of the invention are administered to a patient susceptible to or otherwise at risk of a particular infection. Such an amount is defined to be a "prophylactically effective amount or dose." In this use, the precise amounts again depend on the patient's state of health, weight, and the like.
Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary.
Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved condition is retained. When the symptoms have been alleviated to the desired level, treatment can cease. Patients can, however, require intermittent treatment on a long-term basis upon any recurrence of the disease symptoms.
In general, a suitable effective dose of the compound of the invention will be in the range of 0.1 to 1000 milligram (mg) per recipient per day, preferably in the range of 1 to 100 mg per day. The desired dosage is preferably presented in one, two, three, four or more subdoses administered at appropriate intervals throughout the day. These subdoses can be administered as unit dosage forms, for example, containing 5 to 1000 mg, preferably 10 to 100 mg of active ingredient per unit dosage form. Preferably, the compounds of the invention will be administered in amounts of between about 2.0 mg/kg to 250 mg/kg of patient body weight, between about one to four times per day.
While it is possible to administer the active ingredient of this invention alone, it is preferable to present it as part of a pharmaceutical formulation. The formulations of the present invention comprise at least one compound or inhibitor of this invention in a therapeutically or pharmaceutically effective dose together with one or more pharmaceutically or therapeutically acceptable carriers. Solid carriers inlcude, e.g., starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, and kaolin, and optionally other therapeutic ingredients. Liquid carriers include, e.g., sterile water, polyethylene glycols, non-ionic surfactants, and edible oils such as corn, peanut and sesame oils. In addition, various adjuvants such as are commonly used in the art may be included. For example: flavoring agents, coloring agents, preservatives, and antioxidants, e.g., vitamin E, ascorbic acid, BHT and BHA. Various other considerations are described, e.g., in Gilman et al . (eds) (1990) Goodman and Gilman' s: The Pharmacological Basis of Therapeutics, 8th Ed., Pergamon Press; and Remington's supra . Methods for administration are discussed therein, e.g., for oral, intravenous, intraperitoneal, or intramuscular administration, and others. Pharmaceutically acceptable carriers will include water, saline, buffers, and other compounds described, e.g., in the MERCK INDEX, Merck & Co., Rahway, NJ. Generally, preferred routes of administration are intravenous and intraperitoneal.
These pharmacological agents can be in a variety of forms. These include, for example, solid, semi-solid and liquid dosage forms, such as tablets, pills, powders, liquid solutions or suspensions, liposomes, injectable and infusible solutions. The preferred form depends on the intended mode of administration and therapeutic application. Generally, a pharmacologically acceptable salt of the compound will be used to simplify preparation of the composition. Preferred salts include sodium, potassium, arginine, glycine, alanine, threonine. These are prepared, preferably, in water suitably mixed with a surfactant such as hydroxypropylcellulose. Depending on the specific conditions being treated, such agents may be formulated and administered ■ systemically or locally. Techniques for formulation and administration may be found in Remington's Pharmaceutical Sciences, 18th ed. , Mack Publishing Co., Easton, PA (1990) . Suitable routes may include oral, rectal, transdermal, vaginal, transmucosal, or intestinal administration; parenteral delivery, including intramuscular subcutaneous, intramedullary injections, as well an intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections, just to name a few.
For injection, the agents of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiological saline buffer. For such transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added.
IV. Biological Activity
In Vi tro Antibacterial Evaluation The compounds of the invention were evaluated against several β-lactam resistant bacteria strains by determining the minimum inhibitory concentration (MIC, μg/ml) of each compound with respect to each strain. The MIC, the lowest concentration of antibiotic which inhibits growth of the test organism, was determined by the agar dilution method.
To determine the MIC for bacterial isolates, the test compound was incorporated in a series of two-fold dilutions into liquified Mueller-Hinton agar. Upon solidification, a number of different bacterial strains were spot inoculated with a replicating device onto the agar surface. After overnight incubation, the MIC breakpoint was determined as the lowest drug concentration that completely inhibited growth, disregarding a single colony or a faint haze. The procedures used in these stuides have been standardized by the National Committee for Clinical Laboratory Standards (NCCLS) , as per the NCCLS publication entitled "Methods for Dilution Antimicrobial Susceptibility Tests" (1991) , which is incorporated herein by reference. Aliquots of antimicrobial agents were prepared in phosphate buffered saline (PBS) at pH 7.2. Tween 20 or DMSO was used as a solubilizing vehicle as needed. Standard methods of vortexing, sonicating and gentle heat were used to facilitate solubilizing the test agent. Typically, the concentration of the stock solution was 10X that of the highest drug concentration tested. A 1.28 mg/mL stock solution was used with a subsequent highest working concentration of 128 μg/mL. Serial two-fold dilutions were done through <0.25μg/mL. Each drug level was tested in duplicate. Two-fold drug dilutions were done in sterile 50 mL tubes with a final drug volume of 5 mL. Upon the addition of 45 mL of molten agar, a 10-fold dilution resulted. Two, 25 mL. plates were then poured into 15x150 mm square Petri plates with grids and allowed to harden. A control plate with a reference drug, either cefotaxime, vancomycin or imipenem, was used as the positive growth control. Stock concentrations of reference antibiotics were prepared and frozen at -80°C. Upon preparation, the control plates were sealed and stored in the refrigerator for up to 1 week prior to use; however, imipenem control plates had to be prepared just prior to use. All test plates were used within 24 hours of preparation.
Satisfactory results were obtained where the inoculum contained about 104 colony forming units (cfu) ±
0.5 logs. Starting with pure cultures of the test isolates on agar plates, a few isolated colonies were transferred to a tube of nutrient broth and allowed to grow 4-6 hours at 35-36°C to reach log-phase growth. Dropwise addition of the broth culture to PBS was done to match a 0.5 McFarland turbidity standard equal to 108 cfu/mL. This was further diluted ten-fold in PBS to reach a working inoculum concentration of 107 cfu/mL. When lμL of the working inoculum was applied to the agar surface a concentration of about 104 cfu per spot was obtained.
Disposable sterile lμL loops were used to inoculate test plates, with each isolate in a designated grid on the agar plate. An alternate method of inoculation involved the use of a replica plater, a device with 48 steel pins allowing the simultaneous inoculation of multiple isolates. After the spots had dried, the plates were incubated at 35-36°C for 16-20 hours. Endpoints were assessed as the minimum inhibitory concentration (MIC) of antimicrobial agent. These results are shown in Table I below. The novel agents of this invention are notable for their enhanced activity against S . aureus Col and Enterococci (E. faecium and E. faecalis) . The S. aureus Col strain is a high-level PBP2a producer, whereas S. aureus Col 8A, its isogenic partner, lacks PBP2a. Certain compounds, such as 6 ( (72?) -7- [ (Phenylacetyl) amino] -3- (o-iodophenylthio) -3-cephem- 4-carboxylic acid) and 9
( (72?) -7- [ (Phenylacetyl) amino] -3- (2-biphenylthio) -3-cephem-4- carboxylic acid) , show broad activity against both S. aureus Col and S . aureus Col 8A, as well as Enterococci. The S . aureus Col 8A strain was highly responsive to all test agents including the Cefotaxime control. Thus, the compounds of the present invention are effective against PBP2a-producing bacteria. Compound 12, while lacking activity against S . aureus Col, shows potent activity against enterococci . Certain other compounds of the present invention, such as 22
( (72?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido ] -3- (o-iodophenylthio) -3-cephem-4-carboxylic acid) , are effective against E. coli in addition to Gram-positive organisms. Table 1. Antimicrobial Properties of 7-f AcvlamidoV3- (arvlthiokephems
Organism Cefotax. 1 2 3 4 5 6
S. aureus ATCC 29213 2 0.03 0.03 0.03 0.125 0.03 <0.015
S. aureus ATCC 25923 <1 <0.015 <0.015 <0.015 <0.06 O.015 <0.015
S. epidermidisAΥCC 31432 2 <0.015 <0.015 <0.015 <0.06 <0.015 <0.015
S. aureus Col (MethR)(lac") >32 16 2 2 1-2 2 1
S. aureus 76 (MethR)(lac+) >32 32 16 16 4 16 1
S. aureus 2 0.03 0.03 0.015 <0.06 0.03 <0.015
Col8A(Meths)(lac-)
5. haemolyticus (MethR) >32 >32 16 16 16 16 4
E.faecium ATCC 35667 >32 8 2 4 2 4 0.5
E. faecalis ATCC 29212 >32 2 2 2 4 2 0.5
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32 . aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 . maltophilia ATCC >32 >32 >32 >32 >32 >32 >32
13637
Table 1 fcont . Antimicrobial Properties of 7-(Acvlamido 3-('arvlthioϊceplιems
Organism Cefotax. 7 8 9 M il 12
S. aureus ATCC 29213 2 <0.06 <0.125 0.25 0.125 0.125 <0.06
S. aureus ATCC 25923 <1 <0.06 <0.125 0.125 O.06 <0.015 <0.06
S. epidermidisAΥCC 31432 2 <0.06 <0.125 0.125 <0.06 0.03 <0.06
S. aureus Col (MethR)(lac-) >32 4 4 4 8 1 >32
5. aureus 76 (MethR)(lac+) >32 16 >32 8 32 4 >32
S. aureus Col8A(Meths)(lac") 2 0.06 <0.125 0.25 0.125 0.06 <0.06
S. haemolyticus (MethR) >32 0.125 4-8
E. faecium ATCC 35667 >32 2 4
E.faecalis ATCC 29212 >32 1 4 4 2 2 4
£. co/ ATCC 25922 <1 >32 >32 >32 >32 >32 >32 . aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 . maltophilia ATCC 13637 >32 >32 >32 >32 >32 >32 >32
Table 1 (confl. Antimicrobial Properties of 7-f AcvlamidoV3-(arvIthι olcephems
Organism Cefotax. 11 14 15 12 18
S. aureus ATCC 29213 2 0.06 0.25 0.06 <0.5 <0.25 <0.25
S. aureus ATCC 25923 <1 O.015 0.06 0.03
S. epidermidisAΥCC 31432 2 0.03 0.06 0.03 <0.5 <0.25 <0.25 S. aureus Col (MethR)(lac") >32 4 4 <0.5 2 4 S. aureus 76 (MethR)(lac+) >32 16 4 8 4 4 16 5. aureus Col8A(Meths)(lac ) 2 0.06 0.25 0.06 <0.5 <0.25 <0.25 5. haemolyticus (MethR) >32 16 8 16 16 8 32 E.faecium ATCC 35667 >32 4 1 2 4 1 4
E.faecalis ATCC 29212 >32 2 2 2 E. coli ATCC 25922 <1 >32 >32 >32 >16 >64 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 maltophilia ATCC 13637 >32 >32 >32 >32
SUBSTITUTE SHEET (RULE 26V Table 1 (confl. Antimicrobial Properties of 7-(AcvlamidoV ■3-(arvlthiokephems
Organism Cefotax. 19 20 21 22 23 24
S. aureus ATCC 29213 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus ATCC 25923 <1
S. epidermidisAΥCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac") >32 4 0.5 1 2 2 1
S. aureus 76 (MethR)(lac+) >32 16 4 4 4 8 4
S. aureus Col8A(Meths)(lac-) 2 <0.25 <0.25 <0.25 0.5 <0.25 <0.25
S. haemolyticus (MethR) >32 32 8 32 4 16 8
E. faecium ATCC 35667 >32 2 2 4 2 2 2
E.faecalis ATCC 29212 >32
E. coli ATCC 25922 <1 >32 >32 >32 4 >32 >32
P. aeruginosa ATCC 27853 8-16
X. maltophilia ATCC 13637 >32
Table 1 fcon l. Antimicrobial Properties of 7-f AcvlamidoV3-('arvIth lo ephems
Organism Cefotax. 25 26 27 28 29 30
S. aureus ATCC 29213 2 0.125 <0.25 <0.25 <0.25 <0.25 <0.25
5. aureus ATCC 25923 <1
5. epidermidisAΥCC 31432 2 <0.06 <0.25 <0.25 <0.25 <0.25 <0.25
5. t/reMj Col (MethR)(lac ) >32 1 0.5 1 1 1 0.5
S. aureus 16 (MethR)(lac+) >32 4 4 4 4 4 2
S. aureus Col8A(Meths)(lac") 2 0.125 <0.25 <0.25 <0.25 <0.25 <0.25
S. haemolyticus (MethR) >32 16 8 16 8 16 4
E.faecium ATCC 35667 >32 2 2 1 2 1 1
E. faecalis ATCC 29212 >32
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa ATCC 27853 8-16
-Y. maltophilia ATCC 13637 >32
Table 1 fconfl. Antimicrobial Properties of 7-fAcvlamidoV3-(arvlthio ephems
Organism Cefotax. 31 32 33 34 35 36
S. aureus AYCC 29213 2 <0.25 <0.25 <0.25
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac ) >32 1 1 1 2 0.5 2
5. aureus 76 (MethR)(lac+) >32 4 4 4 8 2 8
S. aureus Col8A(Meths)(lac") 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. haemolyticus (Meth ) >32 8 16 16 8 8 16
E.faecium ATCC 35667 >32 4 2 1 2 1 4
E. faecalis ATCC 29212 >32 2 1 2
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32
X. maltophilia ATCC 13637 >32 Table 1 ( cont . Antimicrobial Properties of 7-f Acvlamido)-3-(arvlthiokeDhems
Organism Cefotax. 32 38 39 40 41 42
S. aureus ATCC 29213 2
5. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lacv) >32 2 4 0.5 4 1 4
S. aureus 76 (MethR)(lac'f) >32 8 16 4 16 4 8
S. aureus Col8A(Meths)(lac 2 <0.25 <0.25 <0.25 <0.25 <0.25 0.5
S. haemolyticus (MethR) >32 16 32 8 32 16 16
E.faecium ATCC 35667 >32 2 4 2 4 2 2
E.faecalis ATCC 29212 >32 2 4 2 4 4 2
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32
X. maltophilia ATCC 13637 >32
Table 1 (cont). Antimicrobial Properties of 7-(Acvlamido)-3-,arvlth] okephems
Organism Cefotax. 43 44 45 46 47 48
S. aureus ATCC 29213 2
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. αwrewi Col (MethR)(lac") >32 4 2 4 1 1 4
S. aureus 76 (MethR)(lac+) >32 8 4-8 16 8 8 16
S. aureus Col8A(Meths)(lac") 2 <0.25 <0.25 0.5 <0.25 <0.25 0.5
5. haemolyticus (MethR) >32 16 8 32 16 16 16
E.faecium ATCC 35667 >32 4 4 4 2 2 4
E. faecalis ATCC 29212 >32 4 4 4 2 4 4
£. co// ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 maltophilia ATCC 13637 >32
Table 1 (confl. Antimicrobial Properties of 7-fAcvlamidoV3-f arvlthi okenhems
Organism Cefotax. 49 50 51 52 53 54
S. aureus ATCC 29213 2 0.5 0.5 1 <0.25
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25
5. αwrezϋ Col (MethR)(lac") >32 4 2 4 4 4 1
S. αwrewi; 76 (MethR)(lac+) >32 16 8 8 16 8 8
S. aureus Col8A(Meths)(lac-) 2 0.5 <0.25 <0.25 0.5 1 <0.25
S. haemolyticus (MethR) >32 32 8 32 16 8 16
E.faecium ATCC 35667 >32 4 4 4 2 8 4
E.faecalis ATCC 29212 >32 4 4 4 4 8
E. coli ATCC 25922 <1 >32 8 >32 >32 1 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 8 >32 maltophilia ATCC 13637 >32 Table 1 (conf). Antimicrobial Properties of 7-(AcvlamidoV3-(arvlthiokephems
Organism Cefotax. 55 56 57
S. aureus ATCC 29213 2
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25
5. aureus Col (MethR)(lac ) >32 4 1 0.5
S. aureus 76 (MethR)(lac+) >32 16 4 2
S. aureus Col8A(Meths)(lac-) 2 <0.25 <0.25 <0.25
S. haemolyticus (MethR) >32 16 16 4
E.faecium ATCC 35667 >32 8 4 1
E.faecalis ATCC 29212 >32 16 2 2
E. coli ATCC 25922 <1 >32 >32 32
P. aeruginosa ATCC 27853 8-16 >32 >32 32
X. maltophilia ATCC 13637 >32
Table 1. Antimicrobial Properties of 7-(Acvlamido)-3-(arvlthiokeDhems
Organism Cefotax. 58 52 60 61 62 63
S. aureus ATCC 29213 2
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. aureus Col (MethR)(lac) >32 <0.25 2 0.5 4 4 4
S. aureus 76 (MethR)(lac+) >32 4 8 4 16 16 16
S. aureus Col8A(Meths)(lac ) 2 <0.25 <0.25 <0.25 0.5 <0.25 <0.25
S. haemolyticus (MethR) >32 4 16 4-8 >32 32 32
E.faecium ATCC 35667 >32 0.5 4 1 4 4 1
E.faecalis ATCC 29212 >32 1 4 1 4 2 1
£ co// ATCC 25922 <1 32 >32 >32 >32 >32 8
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-(arvlthiokephems
Organism Cefotax. 64 6 66 67 68 69
S. aureus ATCC 29213 2 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac") >32 0.5 8 2 4 8 4
S. aureus 76 (MethR)(lac+) >32 4 32 8 16 16 16
S. aureus Col8A(Meths)(lac") 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. haemolyticus (MethR) >32 8 32 8 16 32 32
E.faecium ATCC 35667 >32 0.5 2 1 2 4 2
E.faecalis ATCC 29212 >32 1 1 4 2 4 2
£. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 Table 1. Antimicrobial Pronerties of 7-( AcvlamidoV3 -farvlthic kephems
Organism Cefotax. 70 71 72 73 74 75
5. aureus ATCC 29213 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. aureus Col (MethR)(la ) >32 4 16 4 4 16 8
5. aureus 76 (MethR)(lac+) >32 16 >32 16 16 32 32
S. aureus Col8A(Meths)(lac") 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
5. haemolyticus (MethR) >32 32 >32 32 32 >32 >32
£ faecium ATCC 35667 >32 2 2 2 1 2 2
E.faecalis ATCC 29212 >32 0.5 1 1 0.5 1 1
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-(arvlthiokephems
Organism Cefotax. 76 77 78 79 80
S. aureus ATCC 29213 2 <0.25 <0.25 0.25 0.06 <0.06
S. aureus ATCC 25923 <1 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25
S. aureus Col (MethR)(lac) >32 2 8 4 4 2
S. aureus 76 (MethR)(lac+) >32 8 32 16 8 8
S. aureus Col8A(Meths)(lac-) 2 <0.25 <0.25 0.25 0.06 <0.06
S. haemolyticus (MethR) >32 32 >32 >32 32 8
E.faecium ATCC 35667 >32 2 2 4 2 2
E.faecalis ATCC 29212 >32 1 1 4 1 1
£ coli ATCC 25922 <1 >32 >32 >32 >32 16
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties < of 7-fAcvlamidoV3- arvlthiokephems
Organism Cefotax. 81 82 83 84 85 86
S. aureus ATCC 29213 2 <0.25 1
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac") >32 2 4 4 2 4 8
S. aureus 76 (MethR)(lac+) >32 4 16 16 8 16 16
S. aureus Col8A(Meths)(lac ) 2 <0.25 <0.25 <0.25 <0.25 <0.25 1
S. haemolyticus (MethR) >32 16 32 32 8 16 16
E. faecium ATCC 35667 >32 1 4 2 1 2 2
E.faecalis ATCC 29212 >32 2 1 1 1 4 4
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 2
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 32 Table 1. Antimicrobial Properties of 7-(Acvlamido)-3-farvlthiokeDhems
Organism Cefotax. 87 89 £0 21 22
5. aureus ATCC 29213 2 0.5 1 <0.25
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25 0.5 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac-) >32 1 4 2 8 4 2
S. aureus 16 (MethR)(lac+) >32 4 16 4 16 16 16
S. aureus Col8A(Meths)(lac ) 2 <0.25 <0.25 0.5 1 <0.25 <0.25
S. haemolyticus (MethR) >32 16 16 8 32 >32 8
E.faecium ATCC 35667 >32 1-2 2 4 16 2 16
E.faecalis ATCC 29212 >32 2 4 2 32 1 16
E. coli ATCC 25922 <1 >32 >32 2 4 >32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 8 16 >32 >32
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-(arvlthiokeDhems
Organism Cefotax. 93 94 95 96 97 98
S. aureus ATCC 29213 2 <0.25 0.25 0.06 0.5
S. aureus ATCC 25923 <1 <0.25 <0.25 <0.25
S. epidermidisAYCC 31432 2 <0.25 <0.25 <0.25
S. aureus Col (MethR)(lac") >32 2 4 4 8 2 2
S. aureus 16 (MethR)(lac+) >32 8 16 8 16 4 8
S. aureus Col8A(Meths)(lac ) 2 <0.25 <0.25 <0.25 0.5 0.125
S. haemolyticus (MethR) >32 8 >32 16 16 8 8
E. faecium ATCC 35667 >32 0.5 2 4 4 1 4
E.faecalis ATCC 29212 >32 <0.25 1 4 1 0.5 1
£ coli ATCC 25922 <1 >32 >32 >32 >32 >32 8
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 16
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-farvlthiokephems
Organism Cefotax. 22 100 101 102 103 104
S. aureus AYCC 29213 2 0.125 <0.06 0.25 0.125 0.25 0.5
S. aureus AYCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lac") >32 4 4 4 4 2 2
S. aureus 16 (MethR)(lac+) >32 16 8 8 16 8 4
S. aureus Col8A(Meths)(la ) 2 0.25 0.125 0.25 0.5
S. haemolyticus (MethR) >32 16 16 16 32 4 4
E.faecium ATCC 35667 >32 1 4 2 4 2 4
E.faecalis ATCC 29212 >32 1 1 1 2 1 2
£ coli ATCC 25922 <1 >32 >32 >32 32 >32 8
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 16 Table 1. Antimicrobial Properties of 7-( AcvlamidoV3 -(arvlthiokephems
Organism Cefotax. 105 106 107 108 109 110
S. aureus AYCC 29213 2 0.25 0.25 0.25 0.5 0.125 0.25
S. aureus AYCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lac") >32 4 1 4 4 4 2
S. aureus 76 (MethR)(lac+) >32 16 8 16 8 16 8
S. aureus Col8A(Meths)(lac-) 2 0.25 0.125 0.25 0.5 0.125 0.25
S. haemolyticus (MethR) >32 32 32 32 16 32 >16
£ faecium ATCC 35667 >32 2 4 8 16 4 2
E.faecalis AYCC 29212 >32 1 4 8 16 4 2
£ co// ATCC 25922 <1 >32 >32 >32 4 >32 >32
P. aeruginosa AYCC 27853 8-16 >32 >32 >32 16 >32 >32
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-farvlthiokeDhems
Organism Cefotax. 111 112 113 114 115 116
S. aureus ATCC 29213 2 0.5 ≤0.06 0.125 0.25 0.25 0.25
S. aureus ATCC 25923 <1
S. epidermidisAYCC 31432 2
S. αwreztf Col (MethR)(lac") >32 8 4 4 4 4 4
5. aureus 76 (MethR)(lac+) >32 16 16 16 16 8 16
5. αwrezw Col8A(Meths)(lac") 2 0.25 ≤0.06 0.125 0.125 0.25 0.125
S. haemolyticus (MethR) >32 32 32 >32 32 32 16
E.faecium AYCC 35667 >32 4 4 4 2 8 2
E.faecalis ATCC 29212 >32 4 4 1 1 4 2
£ co// ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa AYCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-fAcvlamidoV3-ιarvlthiokephems
Organism Cefotax. 117 118 119 120 121 122
S. aureus ATCC 29213 2 0.25 0.25 0.125 0.25 0.5 0.25
S. aureus ATCC 25923 <1
5. epidermidisAYCC 31432 2 5. aureus Col (MethR)(lac >32 8 4 2 8 4 4 S. aureus 76 (MethR)(lac+) >32 16 8 8 16 8 8 5. αt/rettf Col8A(Meths)(lac) 2 0.125 0.125 0.06 0.25 0.5 0.25 5. haemolyticus (MethR) >32 32 32 16 32 16 16 £ faecium ATCC 35667 >32 1 4 1 8 2 4
E.faecalis AYCC 29212 >32 2 4 1 4 0.5-1 4 £ co// ATCC 25922 <1 >32 >32 >32 >32 >32 >32
-°. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 Table 1. Antimicrobial Properties of 7-(AcvlamidoV3 -(arvlthic ikephems
Organism Cefotax. 123 124 125 126 127 128
S. aureus AYCC 29213 2 0.06 0.25 0.5 0.125 ≤0.06 ..0.06
S. aureus ATCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lacr) >32 4 4 4 8 4 8
S. aureus 76 (MethR)(lac+) >32 8 16 8 16 8 16
S. aureus Col8A(Meths)(lac) 2 ≤0.06 0.25 0.5 0.125 ≤0.06 ≤0.06
S. haemolyticus (MethR) >32 16 16 8
E.faecium ATCC 35667 >32 4 1 2 2 ' 1 2
£ faecalis AYCC 29212 >32 2 1 0.25 1 1 2
E. coli ATCC 25922 <1 >32 >32 >32 >32 >32 >32
P. aeruginosa AYCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-(Acvlamido)-3-(arvlthiokephems
Organism Cefotax. 129 130 131 132 133 134
S. aureus AYCC 29213 2 ≤0.06 0.125 ≤0.06 0.125 0.125 0.125
S. aureus AYCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (Meth )(lacr) >32 8 2 1 8 2-4 2
S. aureus 76 ( ethR)(lac+) >32 8 4 4 32 8 8
S. aureus Col8A(Meths)(la ) 2 ≤0.06 0.25 ≤0.06 0.25 0.125 0.125
S. haemolyticus (MethR) >32 16 4 32 >32 16 16
E.faecium ATCC 35667 >32 0.5 1 4 1 1
£ faecalis ATCC 29212 >32 0.25 0.25 0.5 4 1 0.5
E. coli ATCC 25922 <1 2 >32 >32 >32 >32 >32
/>. aeruginosa AYCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-( Acvlamidol-3 -farvlthiokephems
Organism Cefotax. 135 136 137 138 139 140
S. aureus AYCC 29213 2 0.125 0.125 0.125 0.125 0.25 0.125
S. aureus AYCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lac ) >32 8 2 2 2 4 2
S. aureus 76 (MethR)(lac+) >32 16 4 4 8 4 8
S. aureus Col8A(Meths)(lac) 2 ≤0.06 0.06 ≤O.Oό 0.25 ≤0.06
S. haemolyticus (MethR) >32 32 4 16 8 4 8
E.faecium AYCC 35667 >32 2-4 1 0.25 0.5 0.5 1
E.faecalis AYCC 29212 >32 2-4 0.5 0.5 0.5 0.125 1
E. coli ATCC 25922 <1 >32 >32 4 32 1 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32 Table 1. Antimicrobial Properties of 7-(AcvlamidoV3 -farvlthiokephems
Organism Cefotax. 141 142 143 144 145 146
S. aureus AYCC 29213 2 0.25 0.25 0.125 0.125 0.125 ≤0.06
S. aureus AYCC 25923 <1
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lac") >32 2 4 8 4 4 2
S. aureus 76 (MethR)(lac+) >32 8 8 16 8 • 16 8
S. aureus Col8A(Meths)(la ) 2 0.25 0.25 0.125 0.125 0.125 ≤0.06
S. haemolyticus (MethR) >32 8 8 16 8 16 8
£ faecium ATCC 35667 >32 0.5 0.5 0.5 0.5 0.5 1
E.faecalis ATCC 29212 >32 1 1 0.5 0.5 0.5 0.5
E. coli AYCC 25922 <1 4 8 >32 32 >32
P. aeruginosa ATCC 27853 8-16 >32 >32 >32 >32 >32 >32
Table 1. Antimicrobial Properties of 7-(AcvlamidoV3-tarvlthiokeDlιems
Organism Cefotax. 147 148
S. aureus ATCC 29213 2 0.125 0.5
S. aureus ATCC 25923 <ι
S. epidermidisAYCC 31432 2
S. aureus Col (MethR)(lac) >32 2 8
S. aureus 76 (MethR)(lac+) >32 8 16
S. aureus Col8A(Meths)(lac-) 2 0.125 0.5
S. haemolyticus (MethR) >32 8 16
E.faecium ATCC 35667 >32 0.5 1
E.faecalis ATCC 29212 >32 0.25 1
£ coli AYCC 25922 <1 4 32
P. aeruginosa ATCC 27853 8-16 >32 >32
Test Compound Legend
Cefotaxime (IR) -3-Acetoxymethyl-7- [ (Z) -2- (aminothiazol-4-yl) -
2- (methoxyi ino)acetamido] -3-cephem-4-carboxylic acid Cmpd 1 (7i?) -7- [ (Phenylacetyl)amino] -3- (p-fluorophenylthio) -3-ce phem-4-carboxylic acid
Cmpd 2 (71?) -7- [ (Phenylacetyl)amino] -3- (m-iodophenylthio) -3-ceph em-4-carboxylic acid Cmpd 3 (7i?) -7- t (Phenylacetyl)amino] -3- (p-chlorophenylthio) -3-ce phem-4-carboxylic acid Cmpd 4 ( IR) -7- [ (Phenylacetyl) mino] -3- (o-bromophenylthio) -3-cep hem-4-carboxylic acid Cmpd 5 (7i?) -7- [ (Phenylacetyl)amino] -3- (p-bromophenylthio) -3-cep hem-4-carboxylic acid Cmpd 6 ( IR) -7- [ (Phenylacetyl) amino] -3- (o-iodophenylthio) -3-ceph em-4-carboxylic acid Cmpd 7 ( IR) - 1 - [ (Phenylacetyl) amino] -3- (dibenzofuranyl-3-thio) -3
-cephem-4-carboxylic acid Cmpd 8 ( IR) -7- [ (Phenylacetyl)amino] -3- (4-biphenylthio) -3-cephem
-4-carboxylic acid Cmpd 9 ( IR) -7- [ (Phenylacetyl) amino] -3- (2-biphenylthio) -3-cephem
-4-carboxylic acid Cmpd 10 (7R) -7- [ (Phenylacetyl) amino] -3- (3-biphenylthio) -3-cephem -4-carboxylic acid
Cmpd 11 (7-R) - 1 - [ (Phenylacetyl) amino] -3- (2-bromo-4-biphenylthio) -
3-cephem-4-carboxylic acid Cmpd 12 ( IR) -7- [ (Phenylacetyl)amino] -3- (4-biphenylmethylthio) -3- cephem-4-carboxylic acid Cmpd 13 ( IR) -7- [ (Phenylacetyl) amino] -3- (2-fluoro-4-biphenylthio)
-3-cephem-4-carboxylic acid Cmpd 14 ( IR) -7- [ (Phenylacetyl) amino] -3- (4-bromo-2-biphenylthio) -
3-cephem-4-carboxylic acid Cmpd 15 ( IR) -7- [ (Phenylacetyl) amino] -3- (o-chlorophenylthio) -3-ce phem-4-carboxylic acid
Cmpd 16 ( IR) -7- [ (Phenylacetyl)amino] -3- (2, 5-dichlorophenylthio) -
3-cephem-4-carboxylic acid Cmpd 17 ( IR) -7- [ (Cyanoacetyl) amino] -3- (o-iodophenylthio) -3-cephe m-4-carboxylic acid Cmpd 18 ( IR) -7- [ (Phenylacetyl) amino] -3- (3 ,4-dichlorophenylthio) -
3-cephem-4-carboxylic acid Cmpd 19 (7i?) -7- [ (Phenylacetyl) amino] -3- (o-carbomethoxyphenylthio
) -3-cephem-4-carboxylic acid Cmpd 20 ( IR) -7- [ (2-Thienylacetyl) amino] -3- (o-bromophenylthio) -3- cephem-4-carboxylic acid
Cmpd 21 (7-R) -7- [ (Phenylacetyl) amino] -3- (dibenzofuranyl-1-thio) -3 -cephem-4-carboxylic acid Cmpd 22 ( IR) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetam ido] -3- (o-iodophenylthio) -3-cephem-4-carboxylic acid Cmpd 23 ( IR) - 1 - [ (Phenylacetyl) amino] -3- (2- (4-cyanophenyl)phenylt hio) -3-cephem-4-carboxylic acid Cmpd 24 ( IR) - 1 - [ (Phenylacetyl)amino] -3- (2,3-dichlorophenylthio) -
3-cephem-4-carboxylic acid Cmpd 25 ( IR) -7- [ (2-Thienylacetyl) amino] -3- (o-iodophenylthio) -3-c ephem-4-carboxylic acid Cmpd 26 ( IR) -7- [ (2-Thienylacetyl) amino] -3- (2, 5-dichlorophenylthi o) -3-cephem-4-carboxylic acid
Cmpd 27 ( IR) - 1 - [ (Phenylacetyl) amino] -3- (2- (2-furyl)phenylthio) -3
-cephem-4-carboxylic acid Cmpd 28 ( IR) -7- [ (Phenylacetyl) amino] -3- (2,4-dichlorophenylthio) -
3-cephem-4-carboxylic acid Cmpd 29 (IR) -7- [ (p-Methoxyphenylacetyl) amino] -3- (o-iodophenylt i o) -3-cephem-4-carboxylic acid Cmpd 30 ( IR) -7- [ (p-Chlorophenoxyacetyl) amino] -3- (o-iodophenylthi o) -3-cephem-4-carboxylic acid Cmpd 31 (7-R) -7- [ (p-Chlorophenoxyacetyl) amino] -3- (2, 5-dichlorophe nylthio) -3-cephem-4-carboxylic acid
Cmpd 32 (7-R) - 1 - [ (p-Methoxyphenylacetyl) amino] -3- (2, 5-dichlorophe nylthio) -3-cephem-4-carboxylic acid Cmpd 33 (7-R) -7- [ (Phenylacetyl) amino] -3- (2-bromo-5-fluorophenylth io) -3-cephem-4-carboxylic acid Cmpd 34 (7-R) - 1 - [ (Cyanoacetyl) amino] -3- (2-biphenylthio) -3-cephem-
4-carboxylic acid Cmpd 35 (7-R) - 1 - [ (Phenylthioacetyl) amino] -3- (2, 5-dichlorophenylth io) -3-cephem-4-carboxylic acid Cmpd 36 (7-R) - 1 - [ (Phenylacetyl) amino] -3- (2-vinylphenylthio) -3-cep hem-4-carboxylic acid
Cmpd 37 (7-R) -7- [ (Phenylacetyl) amino] -3- (2- (2-thienyl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 38 ( IR) -7- [ (Phenylacetyl) amino] -3- (2-hydroxymethylphenylthi o) -3-cephem-4-carboxylic acid Cmpd 39 ( 7R) -7- [ (Phenylacetyl)amino] -3- (2,4-dibromophenylthio) -3
-cephem-4-carboxylic acid Cmpd 40 (7-R) -7- [ (Phenylacetyl) amino] -3- (4-trifluoromethylphenylt hio) -3-cephem-4-carboxylic acid Cmpd 41 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-cyanophenylthio) -3-cep hem-4-carboxylic acid Cmpd 42 (7-R) - 7 - [ (Z) -2 - (aminothiazol-4-yl) -2- (hydroxyimino) acetam ido] -3- (2-biphenylthio) -3-cephem-4-carboxylic acid
Cmpd 43 (7-R) -7- [ (Phenylacetyl) amino] -3- (2- (2- (4-methoxyphenylmet hyl) thien-5-yl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 44 (7-R) - 7 - [ (Phenylacetyl)amino] -3- (2-methylthiophenylthio) -
3-cephem-4-carboxylic acid Cmpd 45 ( IR) - 7 - [ (Phenylacetyl) amino] -3- (2- (2- (N-methylimidazoliu mmethyl) thien-5-yl) henylthio) -3-cephem-4-carboxylic acid Cmpd 46 (7JR) -7- [ (Phenylacetyl) amino] -3- (2- ert-butylphenylthio) -
3-cephem-4-carboxylic acid Cmpd 47 ( IR) - 1 - [ (Phenylacetyl) amino] -3- (2-iso-propyloxyphenylthi o) -3-cephem-4-carboxylic acid Cmpd 48 ( IR) -7- [ (Phenylacetyl) amino] -3- (2- (3- (N-methylimidazoliu mmethyl) furan-2-yl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 49 ( IR) -7- [ (Phenylacetyl) amino] -3- (4-cyano-2-nitrophenylthi o) -3-cephem-4-carboxylic acid Cmpd 50 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-formylphenylthio) -3-ce phem-4-carboxylic acid Cmpd 51 (7JR) -7- [ (Phenylacetyl) amino] -3- (2,4-dicyanophenylthio) -3
-cephem-4-carboxylic acid Cmpd 52 (7R) -7- [ (Cyanoacetyl) amino] -3- (2-cyanophenylthio) -3-ceph em-4-carboxylic acid
Cmpd 53 (7-R) - 7 - [ (Z) -2 - (aminothiazol-4-yl) -2- (cyclopentyloxyimino
) acetamido] -3- (2-cyanophenylthio) -3-cephem-4-carboxylic acid Cmpd 54 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (1-naphthalenylthio) -3-ce phem-4-carboxylic acid
Cmpd 55 (7-R) -7- [ (Phenylacetyl) amino] -3- (2, 3-dihydro-2, 2-dimethyl benzofuran-7-yl) -3-cephem-4-carboxylic acid Cmpd 56 (7-R) - 7- [ (Phenylacetyl) amino] -3- (5,6,7, 8-tetrahydronaphth alen-1-ylthio) -3-cephem-4-carboxylic acid Cmpd 57 ( 7R) -7- [ (Phenylacetyl)amino] -3- (l-bromo-naphthalen-2-ylt hio) -3-cephem-4-carboxylic acid Cmpd 58 ( 7R) -7- [ (Phenylacetyl) amino] -3- (4-isothiouroniummethyl-2
-iodophenylthio) -3-cephem-4-carboxylic acid Cmpd 59 ( IR) -7- [ (Phenylacetyl)amino] -3- (4- (2-imidazolin-2-ylthio methyl) -2-iodophenylthio) -3-cephem-4-carboxylic acid
Cmpd 60 ( 7R) - 7- [ (Phenylacetyl) amino] -3- (4- (N-amidino) isothiouron iummethyl-2-iodophenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 61 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (4- (N-morpholinoethyl) iso thiouroniummethyl-2-iodophenylthio) -3-cephem-4-carboxyla te, inner salt Cmpd 62 ( 7R) -7- [ (Phenylacetyl) amino] -3- (4-tetramethylisothiouron iummethyl-2-iodophenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 63 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (4-isothiouroniummethylph enylthio) -3-cephem-4-carboxylate, inner salt Cmpd 64 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2-isothiouroniummethylph enylthio) -3-cephem-4-carboxylate, inner salt Cmpd 65 (7-R) -7- [ (Phenylacetyl)amino] -3- (4- (1, 2,4-triazol-5-ylthi omethyl)phenylthio) -3-cephem-4-carboxylic acid
Cmpd 66 (7-R) - 7- [ (Cyanoacetyl) amino] -3- (4-isothiouroniummethyl-2- iodophenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 67 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2- (1, 2,4-triazol-5-ylthi omethyl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 68 (7-R) -7- [ (Phenylacetyl) amino] -3- (2- (2-imidazolin-4-on-2-y
1) thiomethyl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 69 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2-tetramethylisothiouron iummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 70 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2- (pyridin-4-yl) thiometh yl)phenylthio) -3-cephem-4-carboxylic acid
Cmpd 71 (7-R) -7 - [ (Phenylacetyl)amino] -3- (l-N-methyltetrazol-5-yl) thiomethyl) henylthio) -3-cephem-4-carboxylic acid Cmpd 72 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2- (2-imidazolin-2-yl) thi omethyl) henylthio) -3-cephem-4-carboxylic acid Cmpd 73 ( 7R) -7- [ (Phenylacetyl)amino] -3- (2- (N-methylpyridin-4-ium
) thiomethyl)phenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 74 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-N,N' -diethylisothiouro niummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 75 (7-R) -7- [ (Phenylacetyl) amino] -3- (2- (pyrimidine-2-yl) thiom ethyl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 76 ( R) -7- [ (Phenylacetyl) amino] -3- (2-N-methylisothiouronium methylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 77 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2- (2-methyl-l, 3,4-thiadi azol-5-yl) thiomethyl)phenylthio) -3-cephem-4-carboxylic acid Cmpd 78 (7-R) -7- [ (Phenylacetyl)amino] -3- (2-N-acetylamidinothiomet hylphenylthio) -3-cephem-4-carboxylic acid Cmpd 79 ( 7R) - 7 - [ (Phenylacetyl) amino] -3- (2-N-cyclopropylisothiour oniummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 80 (7-R) -7- [ (Phenylacetyl)amino] -3- (3-isothiouroniummethylph enylthio) -3-cephem-4-carboxylate, inner salt Cmpd 81 ( 7R) - 7- [ (Phenylacetyl) amino] -3- (2-phenylpyridin-3-ylthio
) -3-cephem-4-carboxylic acid Cmpd 82 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-phenyl-N-methylpyridin
-3-iumthio) -3-cephem-4-carboxylate Cmpd 83 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-phenyl-N-benzylpyridin -3-iumthio) -3-cephem-4-carboxylate
Cmpd 84 ( 7R) -7- [ (4-Chlorophenylacetyl) amino] -3- (2-phenylpyridin-
3-ylthio) -3-cephem-4-carboxylic acid Cmpd 85 (7-R) - 7 - [ (Cyanoacetyl) amino] -3- (2-phenylpyridin-3-ylthio)
-3-cephem-4-carboxylic acid Cmpd 86 (7-R) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino
) acetamido] -3- (2-phenylpyridin-3-ylthio) -3-cephem-4-carb oxylic acid Cmpd 87 ( 7R) -7- [ (Phenylacetyl) amino] -3- (l-N-phenylpyrazol-5-ylth io) -3-cephem-4-carboxylic acid Cmpd 88 (7-R) -7- [ (Cyanoacetyl) amino] -3- (l-N-phenylpyrazol-5-ylthi o) -3-cephem-4-carboxylic acid Cmpd 89 (7-R) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino
) acetamido] -3- (l-N-phenylpyrazol-5-ylthio) -3-cephem-4-ca rboxylic acid Cmpd 90 ( 7R) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (2-fluoroethyloxyimi no) acetamido] -3- (l-N-phenylpyrazol-5-ylthio) -3-cephem-4- carboxylic acid Cmpd 91 ( 7R) -7- [ (Phenylacetyl) amino] -3- (1-N-phenyl-l,2, 3-triazol
-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 92 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-amino-4-phenyl-thiazol
-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 93 (7-R) -7- [ (Phenylacetyl)amino] -3- (4-phenyl-l, 2, 3-thiadiazo
1-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 94 ( 7R) - 7 - [ (Phenylacetyl) amino] -3- (l-N-phenyltetrazol-5-ylt hio) -3-cephem-4-carboxylic acid
Cmpd 95 ( 7R) - 7- [ (Phenylacetyl)amino] -3- (2-oxazol-5-ylphenylthio)
-3-cephem-4-carboxylic acid Cmpd 96 ( 7R) -7 - [ ( (2-aminomethylphenyl) acetyl) amino] -3- (1-N-pheny lpyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 97 ( 7R) -7- [( (4-isothiouroniummethylphenyl) acetyl) amino] -3- (
2-isothiouroniummethylphenylthio) -3-cephem-4-carboxylate
, trifluoroacetic acid salt Cmpd 98 (7-R) -7- [ (Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (2-fluoro ethyloxyimino) acetamido] -3- (l-N-phenylpyrazol-5-ylthio) - 3-cephem-4-carboxylic acid
Cmpd 99 ( 7R) - 7 - [ ( (4-Hydroxyphenyl) acetyl) amino] -3- (2-isothiouron iummethylphenylthio) -3-cephem-4-carboxylic acid, inner salt Cmpd 100 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2- (2-isothiouronium)ethy lphenylthio) -3-cephem-4-carboxylate, inner salt
Cmpd 101 ( 7R) - 7 - [ ( (2-Aminomethylphenyl) acetyl) amino] -3- (2-isothio uroniummethylphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 102 ( 7R) - 7 - [ (Phenylacetyl)amino] -3- (2-cyanopyridin-3-ylthio) -3-cephem-4-carboxylic acid
Cmpd 103 (7i?) -7- [ (Phenylacetyl) amino] -3- (2- (pyrazol-l-yl)pyridin- 3-ylthio) -3-cephem-4-carboxylic acid Cmpd 104 (7-R) - 7 - [ (Z) -2 - (aminothiazol-4-yl) -2- (3-pentyloxyimino) ac etamido] -3- (l-N-phenylpyrazol-5-ylthio) -3-cephem-4-carbo xylic acid Cmpd 105 (7-R) - 1 - [ (Phenylacetyl) amino] -3- (1-N- (pyridin-2-yl)pyrazo 1-5-ylthio) -3-cephem-4-carboxylic acid
Cmpd 106 ( 7R) -7- [ (methylthioacetyl) amino] -3- (l-N-phenylpyrazol-5- ylthio) -3-cephem-4-carboxylic acid Cmpd 107 ( IR) - 7 - [ (cyclopropylacetyl) amino] -3- (l-N-phenylpyrazol-5
-ylthio) -3-cephem-4-carboxylic acid Cmpd 108 ( IR) -7- [ (Z) -2- (2-aminothiazol-4-yl) -2- (cyclopropylmethyl oxyimino) acetamido] -3- (l-N-phenylpyrazol-5-ylthio) -3-cep hem-4-carboxylic acid Cmpd 109 (7-R) - 1 - [ (hexanoyl)amino] -3- (l-N-phenylpyrazol-5-ylthio) -
3-cephem-4-carboxylic acid Cmpd 110 ( 7R) -7- [ (4-pyridylthioacetyl) amino] -3- (1-N-phenylpyrazol
-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 111 ( IR) -7- [ (2-f rylacetyl) amino] -3- (l-N-phenylpyrazol-5-ylt hio) -3-cephem-4-carboxylic acid Cmpd 112 ( 7R) -7- [ (phenyloxyacetyl)amino] -3- (l-N-phenylpyrazol-5-y lthio) -3-cephem-4-carboxylic acid
Cmpd 113 ( 7R) -7- [ (Phenylacetyl) amino] -3- (1-N- (3-nitrophenyl)pyraz ol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 114 (7-R) - 7 - [ (Phenylacetyl)amino] -3- (1-N- (4-methoxyphenyl)pyr azol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 115 ( IR) - 7- [ (2-dithiolanylcarbonyl) amino] -3- (1-N-phenylpyraz ol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 116 ( IR) - 7 - [ (2-thienylacetyl) amino] -3- (l-N-phenylpyrazol-5-y lthio) -3-cephem-4-carboxylic acid Cmpd 117 (7-R) - 1 - [ (Phenylacetyl) amino] -3- (1-N- (3-methoxyphenyl)pyr azol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 118 (7-R) - 7 - [ (2-methyl-l, 3 , 4-thiadiazol-5-ylthio) acetyl) amino
] -3- (l-N-phenylpyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 119 (7-R) -7- [ (phenylthioacetyl) amino] -3- (l-N-phenylpyrazol-5- ylthio) -3-cephem-4-carboxylic acid Cmpd 120 (7-R) - 7 - [ (2-amino-l, 3 , 4-thiadiazol-5-ylthio) acetyl) amino]
-3- (l-N-phenylpyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 121 ( 7R) -7- [2- (2-furyl) -2- (hydroxyimino) acetamido] -3- (1-N-ph enylpyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 122 (7-R) -7- [ (cyanomethylthioacetyl) amino] -3- (1-N-phenylpyraz ol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 123 ( 7R) -7- [ (Phenylacetyl) amino] -3- (1-N- (4-fluorophenyl)pyra zol-5-ylthio) -3-cephem-4-carboxylic acid
Cmpd 124 ( 7R) -7- [ ( (4-aminomethylphenyl) acetyl) amino] -3- (2-isothio uroniummethylphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 125 ( 7R) -7- [ ( (N,N' -diisopropylisothiouronium) acetyl) amino] -3 - (2-isothiouroniummethylphenylthio) -3-cephem-4-carboxyla te, trifluoroacetic acid salt Cmpd 126 (7-R) - 7 - [ (methylthioacetyl) amino] -3- (2- (N-amidino) isothio uroniummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 127 ( 7R) - 7 - [ (methylthioacetyl) amino] -3- (2-isothiouroniummeth ylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 128 ( 7R) -7- [ (Phenylacetyl) amino] -3- (2-guanidiniummethylpheny lthio) -3-cephem-4-carboxylate, inner salt Cmpd 129 (7-R) - 7 - [ (Z) - 2 - (Aminothiazol-4-yl) -2- (hydroxyimino) acetam ido] -3- (2-isothiouroniummethylphenylthio) -3-cephem-4-car boxylate, inner salt Cmpd 130 (7-R) - 7- [ (Z) -2 - (Phenyl) -2 - (hydroxyimino) acetamido] -3- (1-N
- (phenyl)pyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 131 (7-R) - 7- [( (Z-2-Chlorovinyl) thioacetyl) amino] -3- (1-N-pheny lpyrazol-5-ylthio) -3-cephem-4-carboxylic acid Cmpd 132 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2- (pyrazol-l-yl)phenylth io) -3-cephem-4-carboxylic acid Cmpd 133 ( IR) -7- [ (methylthioacetyl) amino] -3- (2- (N-amino) isothiour oniummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 134 (7-R) - 7 - [ (3-iodoprop-1-ylthioacetyl) amino] -3- (2-isothiour oniummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 135 (7-R) - 7 - [ (Methylthioacetyl) amino] -3 - (2-guanidiniummethylp henylthio) -3-cephem-4-carboxylate, inner salt Cmpd 136 ( IR) - 7- [ (Z) -2- (Phenyl) -2- (hydroxyimino) acetamido] -3- (2-i sothiouroniummethylphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 137 ( IR) -7- [ ( (2-guanidinoethylthio) acetyl) amino] -3- (2-isothi ouroniummethylphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 138 ( IR) -7- [ (phenylacetyl) amino] -3- [2,4- (bis) (isothiouronium methyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
Cmpd 139 (7-R) - 7 - [ (Z) - 2 - (Aminothiazol-4-yl) -2- (hydroxyimino) acetam ido] -3- [2,4- (bis) isothiouroniummethylphenylthio] -3-cephe m-4-carboxylate, trifluoroacetic acid salt Cmpd 140 (7-R) - 7 - [ (methylthioacetyl) amino] -3- [2,4- (bis) isothiouron iummethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 141 (7-R) - 7- [ ( (2-guanidinoethylthio)acetyl) amino] -3- [2,4- (bis
) isothiouroniummethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
Cmpd 142 (7-R) - 7 - [ ( ( 2 - (isothiouronium)ethylthio) acetyl) amino] -3- [2
,4- (bis) isothiouroniummethylphenylthio] -3-cephem-4-carbo xylate, trifluoroacetic acid salt Cmpd 143 ( 7R) -7- [ ( (2- (isothiouronium) ethylthio) acetyl) amino] -3- [2 - (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylat e, trifluoroacetic acid salt Cmpd 144 (7-R) - 7 - [ ( ( 2 - (isothiouronium)propylthio) acetyl) amino] -3- [
2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxyla te, trifluoroacetic acid salt Cmpd 145 (7-R) -7- [( (2- (guanidino) ropylthio) acetyl) amino] -3- [2- (is othiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 146 (7-R) - 7 - [ (Phenylacetyl) amino] -3- (2-amidinomethylthiomethy lphenylthio) -3-cephem-4-carboxylate, inner salt Cmpd 147 (7-R) - 7- [( (2-guanidinoethylthio) acetyl) amino] -3- (2-amidin omethylthiomethylphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt Cmpd 148 (7-R) - 7 - [ ( (2-guanidinoethylthio) acetyl) amino] -3- [2- (2-dim ethylaminoethyl) thiomethylphenylthio] -3-cephem-4- carboxylate, trifluoroacetic acid salt
In Vivo Antibacterial Evaluation
Compounds with superior activity in vi tro when compared to reference antibiotics, are further evaluated in a murine model for lethal bacteremic peritonitis.
Groups of 5 female Swiss-Webster mice (Simonsen, Gilroy, CA) each are challenged by the intraperitoneal (IP) route with tenfold increments of a bacterial inoculum. This permits calculation of the mean lethal dose (LD50) and the LD100. For preliminary evaluation of a new antibiotic, mice are challenged IP with an LD100 titer of bacteria. In two equal doses administered at the time of bacterial challenge and 2 hours later, groups of 10 mice each are treated subcutaneously with two-fold increments of the test drug and an antibiotic of known efficacy in mice and humans (i.e., positive control) . Mice are observed for 72h. Those alive at 72h are considered long term survivors. The total drug dose in mg/kg that protects 50% of mice in a group from death is termed the mean protective dose (PD50) . PD50s are similarly determined for several pathogens. The quantitative endpoints for the new drug are then compared with those obtained with reference antibiotics. Six ten-fold dilutions of inoculum suspended in 0.5 mL of sterilized 7% hog gastric mucin (Sigma) are injected IP in groups of 5 mice each. A control group of 5 mice receive mucin alone. Mice are observed for 72h. Those alive at 72h are considered long term survivors. The mean lethal dose (LD50) and 100% lethal dose (LD100) are determined by the probit test.
For antibiotic efficacy studies, mice are challenged IP with bacterial titers that will afford an LD100 for the test strain. In two equal doses administered at the time of bacterial challenge and 2 hours later, groups of 10 mice each are treated by the subcutaneous route (SC) with twofold increments of the test antibiotic; another group is treated similarly with a reference antibiotic of known efficacy in animals and man. Drug doses can range from 0.01 to 512 mg/kg. If the drug is poorly soluble, Tween 20 or propylene glycol will be employed to solubilize the drug. Animals are observed for 72h. The 50% protective dose (PD50) is calculated in mg/kg by the probit method. The PD50 is the same as the 50% effective dose (ED50) and the 50% curative dose (CD50) . Samples of blood from the hearts of all animals that die and from half the mice that survive are cultured on brain-heart infusion agar. Animals that received a protective dosage of the test drug will be alive at 72h, although they may appear moderately ill to very ill during the observation period. Infected, placebo-treated control mice and those receiving non-effective i.e. lower dosages of the test drug will demonstrate a high rate of mortality. Most of these mice will die within 6 to 48 h. Those alive at 72h will be considered long term survivors.
(Compound numbers correspond to those in MIC tables,- data reported for challenge with methicillin-susceptible S. aureus strain ATCC 13709. )
Mortality
Compound : Vancomycin 17 81 81
Dose
16 mg/kg 0/10 0/10 0/10 2/10 0/10
8 mg/kg 0/10 1/10 0/10 5/10 0/10
4 mg/kg 2/10 5/10 5/10 10/10 3/10
2 mg/kg 8/10 8/10 8/10 8/10 4/10
ED50 (mg/kg) 2.81 3.65 3.41 8.46 1.89
Challenge str MIC 1 < 0.5 0.125 0.125 0.06
(μg/ml)
Mortal. Lty
Compound : Vancomγcin Cefotaxime 6 41 8j3
Dose
40 mg/kg 1/10
20 mg/kg 4/10
10 mg/kg 2/10 3/10 1/10 0/10 0/10
5 mg/kg 5/10 5/10 1/10 0/10 0/10
2.5 mg/kg 5/10 8/10 4/10 0/10 0/10
1.25 mg/kg 6/10 4/10 6/10 1/10
0.625 mg/kg 8/10 9/10 8/10 2/10
0.3125 mg/kg
ED50 (mg/kg) 2.79 7.47 1.57 1.13 0.31
95% conf . interval 0.79- 2.91- 0.83- 0.81- 0.12-
4.80 12.04 2.31 1.45 0.74
Challenge str MIC 1 2 0.03 0.06 0.25
(μg/ml)
Mortal: Lty
Compound : 81 92 101 103 104
Dose
40 mg/kg
20 mg/kg
10 mg/kg 8/10 0/10 0/10 9/10
5 mg/kg 8/10 0/10 0/10 5/10 8/10
2.5 mg/kg 9/10 2/10 0/10 7/10 8/10
1.25 mg/kg 9/10 3/10 2/10 7/10 10/10
0.625 mg/kg 9/10 5/10 5/10 7/10 6/10
0.3125 mg/kg 8/10
ED50 (mg/kg) >10 0.74 0.65 2.67 >10
95% conf . interval -- 0.39- 0.36- 1.13- --
1.08 0.94 4.21
Challenge str MIC 0.5 0.062 0.125 0.125 0.5
(μg/ml)
Mortal: Lty
Compound : 102 106 115 108 110
Dose
40 mg/kg
20 mg/kg
10 mg/kg 0/10 10/10 0/10
5 mg/kg 0/10 4/10 10/10 6/10
2.5 mg/kg 5/10 1/10 7/10 9/10 8/10
1.25 mg/kg 7/10 2/10 9/10 8/10 7/10
0.625 mg/kg 8/10 6/10 8/10 9/10 8/10
0.3125 mg/kg 10/10 9/10
0.1562 mg/kg 9/10
ED50 (mg/kg) 1.75 0.72 3.04 >10 3.45
95% conf. interval 1.12- 0.44- 1.73- -- 1.58-
2.38 1.00 4.34 5.32
Challenge str MIC 0.125 0.125 0.25 0.5 0.25
(μg/ml) Mortality
Compound : 107 118 119 122 125
Dose
40 mg/kg
20 mg/kg
10 mg/kg 0/10 2/10 0/10 7/10
5 mg/kg 0/10 4/10 8/10 0/10 4/9
2.5 mg/kg 5/10 8/10 7/10 0/10 7/10
1.25 mg/kg 6/10 6/10 10/10 6/10 10/10
0.625 mg/kg 8/10 8/10 9/10 8/10 9/10
0.3125 mg/kg 10/10
0.1562 mg/kg
ED50 (mg/kg) 1.58 3.38 >5 1.13 >10
95% conf. interval 0.99- 1.10- -- 0.81- --
2.17 5.66 1.45
Challenge str MIC 0.125 0.06 0.25 0.125 --
(μg/ml)
Mortal ity
Compound : 127 130 131 137
Dose
40 mg/kg
20 mg/kg
10 mg/kg 6/10 0/10
5 mg/kg 1/10 6/10 0/10
2.5 mg/kg 2/10 9/10 4/10
1.25 mg/kg 4/10 7/10 7/10
0.625 mg/kg 5/10 10/10 9/10 6/10
0.3125 mg/kg 6/10 5/10
0.1562 mg/kg 7/10
0.0781 mg/kg 9/10
0.0391 mg/kg 8/10
EDS0 (mg/kg) 0.59 >10 1.73 0.75
95% conf. interval 0.11-1.07 -- 1.18-2 27 0-1.75
Challenge str MIC -- 0.125 0.06 0.06
(μg/ml)
Mortality
Compound: 127 130 131 137 138 Dose 40 mg/kg
20 mg/kg
10 mg/kg 6/10 0/10
5 mg/kg 1/10 6/10 0/10
2.5 mg/kg 2/10 9/10 4/10 0/10
1.25 mg/kg 4/10 7/10 7/10 0/10
0.625 mg/kg 5/10 10/10 9/10 1/10
0.3125 mg/kg 6/10 8/10 0/10
0.1562 mg/kg 9/10 0/10
0.0781 mg/kg 6/10
10 0.0391 mg/kg 10/10
0.0195 mg/kg 9/10
ED50 (mg/kg) 0.59 >10 1.73 0.38 0.075
95% conf. interval 0.11-1.07 -- 1.18-2.27 0.27-0.48 0.053-
0.096
Challenge str MIC -- 0.125 0.06 0.125 --
15 (μg/ml)
Mortali ty
Compound : 139 140 141 142 143
2.0 Dose
40 mg/kg
20 mg/kg
10 mg/kg
5 mg/kg
25 2.5 mg/kg 1/10 0/10 1/10 0/10
1.25 mg/kg 0/10 0/10 1/10 0/10
0.625 mg/kg 4/10 2/10 2/10 0/10 2/10
0.3125 mg/kg 8/10 1/10 3/10 1/10 6/10
0.1562 mg/kg 4/10 0/10 3/10 3/10 9/10
30 0.0781 mg/kg 8/10
0.0391 mg/kg 9/10
ED50 (mg/kg) 0.28 <0.156 <0.156 <0.156 0.34
95% conf. interval 0.084-0.47 -- -- -- 0.15-0.53
Challenge str MIC -- -- -- -- --
35 (μg/ml)
Mortality
Compound : 144 145 147 Dose 40 mg/kg
20 mg/kg
10 mg/kg
5 mg/kg
2.5 mg/kg 0/10 0/10
1.25 mg/kg 0/10 0/10
0.625 mg/kg 3/10 3/10 0/10
0.3125 mg/kg 6/10 7/10 0/10
0.1562 mg/kg 8/10 9/10 0/10
0.0781 mg/kg 7/10
0.0391 mg/kg 9/10
ED50 (mg/kg) 0.35 0.40 0.081
95% conf. interval 0.23-0.47 0.28-0.53 0.06-0.10
Challenge str MIC -- -- --
(μg/ml)
EXAMPLES The present invention will be more fully described in conjunction with the following specific examples which are not to be construed in any way as limiting the scope of the invention.
EXAMPLE 1 (7-R) -7- t (phenylacetyl) amino] -3- (4-biphenylthio) -3-cephem-4-c arboxylate, benzhydryl ester
To a stirred solution of (71?) -7- [ (phenylacetyl) amino] -3-trifluoromethanesulfonyloxy-3-cephem-4-carboxylate, benzhydryl ester (760 mg, 1.2 mmol) in dry DMF (12 mL) at -60° under nitrogen was added a solution of sodium 4-phenylbenzenethiolate in DMF (3 mL) , prepared from 4-phenylbenzenethiol (320 mg, 1.56 mmol) and sodium hexamethyldisilazide (1.2 mL of a 1M solution in THF, 1.2 mmol) . After 25 min. the reaction was quenched by addition of aqueous 10% ammonium chloride solution (1 mL) and water (120 mL) . The resulting milky solution was extracted with dichloro-methane (3 x 50 mL) and 10% EtOAc/hexane (3 x 50 mL) . The organic extracts were dried over sodium sulfate and filtered, and the solvent was removed with a rotary evaporator. The crude product was purified by chromatography on silica gel (chromatotron, 4 mm plate, 5% EtOAc/hexane, then 0.25% MeOH/ dichloromethane) to afford 450 mg (56%). of the title compound. 1H NMR (CDC13) d 3.17 (d, IH, J=18) , 3.31 (d, IH, J=17) , 3.63 (d, IH, J=16) , 3.69 (d, IH, J=16) , 5.01 (d, IH, J=4) , 5.79 (dd, IH, J=4,9) , 6.06 (d, IH, J=9) , 7.00 (s, IH) , 7.2-7.7 (m, 25H) .
EXAMPLE 2
(7JR) -7- [ (phenylacetyl) amino] -3- (4-bromophenylthio) -3-cephem-
4-carboxylate, benzhydryl ester To a well-stirred suspension of (71?) -7-
[ (phenylacetyl) amino] -3-trifluoromethanesulfonyloxy-3- cephem-4-carboxylate, benzyhydryl ester (190 mg, 0.30 mmol) in dry THF (20 mL) at -78° under nitrogen was added a suspension of sodium 4-bromobenzenethiolate in THF (5 mL) , prepared from 4-phenylbenzenethiol (62 mg, 0.33 mmol) and sodium hexamethyldisilazide (0.30 mL of a 1M solution in THF, 0.30 mmol) . After 1 h, the reaction was quenched by addition of aqueous 5% ammonium chloride solution (3 mL) . The resulting mixture was extracted with dichloromethane (3 x 25 mL) , and the extracts were washed with water and saturated sodium chloride solution. The organic phase was dried over sodium sulfate and filtered, and the solvent was removed with a rotary evaporator. The crude product was purified by chromatography on silica gel (chromatotron, 2 mm plate, 0.25% MeOH/dichloromethane) to afford 85 mg (42%) of the title compound. XH NMR (CDC13) d 3.10 (d, IH, J=18) , 3.28 (d, IH, J=17) , 3.63 (d, IH, J=16) , 3.69 (d, IH, J=16) , 5.00 (d, IH, J=4) , 5.81 (dd, IH, J=4,9) , 6.07 (d, IH, J=9) , 6.98 (s, IH) , 7.2-7.5 (m, 20H) .
Substantially following the methods described in Examples 1 - 2, the compounds of the invention listed below in Examples 3 - 14 were obtained.
EXAMPLE 3
(7i?) -7- [ (phenylacetyl)amino] -3- (4-iodophenylthio) -3-cephem-4 -carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium 4-iodo-phenylthiolate was used.
EXAMPLE 4 (71?) -7- [ (phenylacetyl)amino] -3- [2-bromo-4- ( henyl) henylthio] -3-cephem-4-carboxylate, benzhydryl ester This was prepared by a procedure similar to that for Example 1, but sodium 2-bromo-4- (phenyl)phenylthiolate was used.
EXAMPLE 5 (7-R) -7- [ (phenylacetyl)amino] -3- [2-phenyl-4-bromophenylthio] - 3-cephem-4-carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium 2-phenyl-4-bromophenylthiolate was used. EXAMPLE 6
(71?) -7- [ (phenylacetyl) amino] -3- (3-iodophenylthio) -3-cephem-4
-carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium 3-iodo-phenylthiolate was used.
EXAMPLE 7 (71?) -7- [ (phenylacetyl)amino] -3- (2,4-dichlorophenylthio) -3-ce phem-4-carboxylate, benzhydryl ester This was prepared by a procedure similar to that for Example 1, but sodium 2,4-dichlorophenylthiolate was used.
EXAMPLE 8 (71?) -7- [ (phenylacetyl)amino] -3- [4- (2-imidazolyl)phenylthio] - 3-cephem-4-carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium 4- (2-imidazolyl)phenylthiolate was used.
EXAMPLE 9
(71?) -7- [ (phenylacetyl) amino] -3- (dibenzofuran-3-ylthio) -3-cep hem-4-carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium dibenzofuran-3-thiolate was used.
EXAMPLE 10 (71?) - [ (phenylacetyl) amino] -3- (dibenzofuranyl-2-ylthio) -3-cep hem-4-carboxylate, benzhydryl ester This was prepared by a procedure similar to that for Example 1, but sodium dibenzofuran-2-thiolate was used.
EXAMPLE 11
(71?) -7- [ (phenylacetyl) amino] -3- (dibenzofuran-1-ylthio) -3-cep hem-4-carboxylate, benzhydryl ester This was prepared by a procedure similar to that for Example 1, but sodium dibenzofuran-1-thiolate was used.
EXAMPLE 12 (71?) -7- [ (phenylacetyl)amino] -3- (dibenzofuran-4-ylthio) -3-cep hem-4-carboxylate, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium dibenzofuran-4-thiolate was used.
EXAMPLE 13 (71?) -7- [ (phenylacetyl)amino] -3- [6-
(hydroxymethyl)dibenzofuran-3-ylthio] -3-cephem-4-carboxylate
, benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium
SSSD/14615. v01 6- (hydroxymethyl) dibenzofuran-3-thiolate was used.
EXAMPLE 14 (71?) -7- [ (phenylacetyl) amino] -3- [7- (hydroxymethyl)dibenzof ran-3-ylthio] -3-cephem-4-carboxylate , benzhydryl ester
This was prepared by a procedure similar to that for Example 1, but sodium 7- (hydroxymethyl) dibenzofuran-3-thiolate was used.
EXAMPLE 15
(71?) -7- [ (phenylacetyl)amino] -3- (2-biphenylthio) -3-cephem-4-c arboxylate, 4-methoxybenzyl ester
To a stirred solution of (71?) -7- [ (phenylacetyl) amino] -3-trifluoromethanesulfonyloxy-3 -cephem-4-carboxylate, 4-methoxybenzyl ester (700 mg, 1.2 mmol) in dry DMF (12 mL) at -60° under nitrogen was added a solution of sodium 2-phenylbenzenethiolate in DMF (3 mL) , prepared from 2-phenylbenzenethiol (320 mg, 1.56 mmol) and sodium hexamethyldisilazide (1.2 mL of a 1M solution in THF, 1.2 mmol) . After 25 min. the reaction was quenched by addition of aqueous 10% ammonium chloride solution (1 mL) and water (120 mL) . The resulting milky solution was extracted with dichloro-methane (3 x 50 mL) and 10% EtOAc/hexane (3 x 50 mL) . The organic extracts were dried over sodium sulfate and filtered, and the solvent was removed with a rotary evaporator. The crude product was purified by chromatography on silica gel (chroma-totron, 4 mm plate, 5% EtOAc/hexane, then 0.25% MeOH/dichloromethane) to afford 450 mg (56%) of the title compound. Substantially following the methods described in Example 15, the compounds of the invention listed below in Examples 16 - 32 were obtained.
EXAMPLE 16
(71?) -7- [ (phenylacetyl)amino] -3- [2- (4-cyanophenyl)phenylthio]
-3-cephem-4-carboxylate, 4-methoxybenzyl . ester
This was prepared by a procedure similar to that for Example 15, but sodium 2- (4-cyanophenyl)phenylthiolate was used.
EXAMPLE 17
(71?) -7- [ (phenylacetyl)amino] -3- [2,3-dichlorophenylthio] -3-ce phem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 2, 3-dichlorophenylthiolate was used.
EXAMPLE 18 (71?) -7- [ (phenylacetyl) amino] -3- [2,5-dichlorophenylthio] -3-ce phem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 2, 5-dichlorophenylthiolate was used. EXAMPLE 19
(71?) -7- [ (phenylacetyl)amino] -3- (2-bromo-5-fluorophenylthio) -
3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 2-bromo-5-fluorophenylthiolate was used.
EXAMPLE 20
(71?) -7- [ (phenylacetyl)amino] -3- (2-bromophenylthio) -3-cephem-
4-carboxylate, 4-methoxybenzyl ester This was prepared by a procedure similar to that for Example 15, but sodium 2-bromophenylthiolate was used.
EXAMPLE 21 (71?) -7- [ (phenylacetyl)amino] -3- (4-chlorophenylthio) -3-cephem -4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 4-chlorophenylthiolate was used.
EXAMPLE 22 (71?) -7- [ (phenylacetyl)amino] -3- [4- (ethoxycarbonyl)phenylthio ] - 3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 4- (ethoxycarbonyl)phenyl thiolate was used. EXAMPLE 23
(71?) -7- [ (phenylacetyl) amino] -3- [2- (ethoxycarbonyl)phenylthio
]- 3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 2- (ethoxycarbonyl)phenyl thiolate was used.
EXAMPLE 24 (71?) -7- [ (phenylacetyl) amino] -3- [2-iodophenylthio] -3-cephem-4 -carboxylate, 4-methoxybenzyl ester This was prepared by a procedure similar to that for Example 15, but sodium 2-iodophenyl thiolate was used.
EXAMPLE 25
(71?) -7- [ (phenylacetyl) amino] -3- (2-chlorophenylthio) -3-cephem
-4-carboxylate, 4-methoxybenzyl ester This was prepared by a procedure similar to that for Example 15, but sodium 2-chlorophenyl thiolate was used.
EXAMPLE 26 (71?) -7- [ (phenylacetyl)amino] -3- [4-bromo-3- (trifluoromethoxy) -phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 4-bromo-3- (trifluoromethoxy)phenyl thiolate was used.
EXAMPLE 27
(71?) -7- [ (phenylacetyl) amino] -3- [ [4- (2-imidazolyl)methyl]phen yl-thio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but potassium 4- [ (2-imidazolyl)methyl] phenyl thiolate was used.
EXAMPLE 28 (71?) -7- [ (phenylacetyl)amino] -3- [ [4- (1-methyl-2-imidazolyl) e thyl] -phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but potassium 4- [ (1-methyl-2-imidazolyl)methyl] phenyl thiolate was used.
EXAMPLE 29
(71?) -7- [ (phenylacetyl)amino] -3- [2-fluoro-4- ( henyl)phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but potassium
2-fluoro-4- (phenyl)phenylthiolate was used.
EXAMPLE 30 (71?) -7- [ (2-Phenylacetyl)amino] -3- [3- (2-thienyl)phenylthio] -3 -cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 3- (2-thienyl)phenyl thiolate was used. EXAMPLE 31
(71?) -7- [ (2-Phenylacetyl) amino] -3- [3 , 4-dichlorophenylthio] -3- cephe -4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but sodium 3 , 4-dichlorophenyl thiolate was used.
EXAMPLE 32 (71?) -7- [ (2-Thienylacetyl)amino] -3- [5-bromo-4- (1-methyl-2-imidazolyl)phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 15, but
(71?) -7- [ (2-thienylacetyl) amino] -3-trifluoromethanesulfonylox y-3-cephem-4-carboxylate, 4-methoxybenzyl ester and sodium 5-bromo-4- (1-methyl-2-imidazolyl) phenyl thiolate was used.
EXAMPLE 33 (71?) -7-Amino-3- (2-bromophenylthio) -3-cephem-4-carboxylate, benzhydryl ester To a solution of
(71?) -7- (phenylacetyl) amino-3- (2-bromophenylthio) -3-cephem-4- carboxylate, benzhydryl ester (812 mg, 1.21 mmol) and pyridine (195 μL, 2.42 mmol) in dichloromethane (5 mL) at 0° was added slowly a solution of phosphorus pentachloride (403 mg, 1.94 mmol) in dichloromethane (9 mL) . After 15 min at 0°, isobutyl alcohol (1.1 mL, 12.1 mmol) was added in one portion. The mixture was diluted with dichloromethane (20 mL) , and was washed with 5% aqueous sodium bicarbonate solution (20 mL) . The organic layer was dried with sodium sulfate, and the solvent was removed with a rotary evaporator. The residue was subjected to flash chromatography on silica gel (33% to 50% ethyl acetate/hexane) to afford 300 mg (45%) of the title compound.
EXAMPLE 34 (71?) -7-Amino-3- (2-iodophenylthio) -3-cephem-4-carboxylate,
4-methoxybenzyl ester A mixture of (71?) -7- (phenylacetyl) amino-3- (2-iodophenylthio) -3-cephem-4-c arboxylate, 4-methoxybenzyl ester (850 mg, 1.21 mmol) and pyridine (195 μL , 2.42 mmol) in dichloromethane (5 mL) at 0° was treated slowly with a solution of phosphorus penta-chloride (403 mg, 1.94 mmol) in dichloromethane (9 mL) . After 15 min at 0°, isobutyl alcohol (1.1 mL, 12.1 mmol) was added in one portion. The mixture was diluted with dichloromethane (20 mL) , and was washed with 5% aqueous sodium bicarbonate solution (20 mL) . The organic layer was dried with sodium sulfate, and the solvent was removed with a rotary evaporator. The residue was subjected to flash chromatography on silica gel (33% to 50% ethyl acetate/hexane) to afford 300 mg (50%) of the title compound.
Substantially following the methods described in Examples 33 - 34, the compounds of the invention listed below in Examples 35 - 50 can be obtained.
EXAMPLE 35 (71?) -7-Amino-3- (4-biphenylthio) -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- (4-biphenylthio) -3-cephem-4-c arboxylate, benzhydryl ester is used.
EXAMPLE 36
(71?) -7-Amino-3- [4- (2-imidazolyl)phenylthio] -3-cephem-4-carbo xylate, benzhydryl ester
The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [4- (2-imidazolyl) phenylthio] - 3
-cephem-4-carboxylate, benzhydryl ester is used.
EXAMPLE 37 (71?) -7-Amino-3- [3-bromo-4- (phenyl)phenylthio] -3-cephem-4-car boxylate, benzhydryl ester
The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [3-bromo-4- (phenyl) phenylthio ] -3-cephem-4-carboxylate, benzhydryl ester is used.
EXAMPLE 38
(71?) -7-Amino-3- [ (2-trifluoromethoxy)phenylthio] -3-cephem-4-c arboxylate, benzhydryl ester
The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [ (2-trifluoromethoxy) phenylth io] -3-cephem-4-carboxylate, benzhydryl ester is used.
EXAMPLE 39 (71?) -7-Amino-3- (dibenzofuran-2-ylthio) -3-cephem-4-carboxylat e, benzhydryl ester The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- (dibenzofuran-2-ylthio) -3-cep h em-4-carboxylate, benzhydryl ester is used.
EXAMPLE 40
(71?) -7-Amino-3- [4- (2-thienyl)phenylthio] -3-cephem-4-carboxyl ate, benzhydryl ester
The procedure of Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [4- (2-thienyl)phenylthio] -3-c ephem-4-carboxylate, benzhydryl ester is used.
EXAMPLE 41
(71?) -7-Amino-3- [ (6-chloromethyl)dibenzofuran-2-ylthio] -3-cep hem-4-carboxylate, 4-methoxybenzyl ester A procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [6- (hydroxymethyl) dibenzofura n-2-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used in the presence of a second equivalent of phosphorus pentachloride and pyridine.
EXAMPLE 42
(71?) -7-Amino-3- [ (7-chloromethyl)dibenzofuran-3-ylthio] -3-cep hem-4-carboxylate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [7- (hydroxymethyl) dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used in the presence of a second equivalent of phosphorus pentachloride and pyridine.
EXAMPLE 43 (71?) -7-Amino-3- (2,5-dichlorophenylthio) -3-cephem-4-carboxyla te, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- (2, 5-dichlorophenylthio) -3-ce phem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 44
(71?) -7-Amino-3- [4- (2-oxazolylmethyl)phenylthio] -3-cephem-4-c arboxylate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [4- (2-oxazolylmethyl)phenylth io] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 45 (71?) -7-Amino-3- [4- (1-imidazolylmethyl)phenylthio] -3-cephem-4
-carboxylate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [4- (1-imidazolylmethyl)phenyl thio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 46
(71?) -7-Amino-3- [4- (1-imidazolylmethy1)phenylthio] -3-cephem-4
-carboxylate, 4-methoxybenzyl ester Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [4- (1-imidazolylmethyl)phenyl thio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 47
(71?) -7-Amino-3- [2-bromo-4- (ethoxycarbonyl)phenylthio] -3-ceph em-4-carboxylate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [2-bromo-4- (ethoxycarbonyl)ph enylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 48 (71?) -7-Amino-3- [4- (2-thiazolyl)phenylthio] -3-cephem-4-carbox ylate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [4- (thiazolyl)phenylthio] -3-c ephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 49 (71?) -7-Amino-3- [4- (cyanamido)phenylthio] -3-cephem-4-carboxyl ate, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except (71?) -7- [ (phenylacetyl) amino] -3- [4- (cyanamido) phenylthio] -3-c ephem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 50 (71?) -7-Amino-3- [4- (2-furyl)phenylthio] -3-cephem-4-carboxylat e, 4-methoxybenzyl ester
Procedure similar to Example 33 is employed, except
(71?) -7- [ (phenylacetyl) amino] -3- [4- (2-furyl) phenylthio] -3-cep hem-4-carboxylate, 4-methoxybenzyl ester is used.
EXAMPLE 51
(71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino) acetyl]
-amino] -3- (4-bromophenylthio) -3-cephem-4-carboxylate, benzhydryl ester. To solution of
2- (2-amino-4-thiazolyl) -2- (syn-methoxyimino) ] acetic acid (40 mg, 0.20 mmol) and 1-hydroxybenzotriazole (27 mg, 0.20 mmol) in DMF (2 mL) was added dicyclohexylcarbodiimide (45 mg, 0.22 mmol) . After stirring for 15 min, a solution of (71?) -7-amino-3- (2-bromophenylthio) -3-cephem-4-carboxylate, benzyhydryl ester (110 mg, 0.20 mmol) in DMF (1.5 mL) was added. The mixture was stirred at room temperature overnight, and was partitioned between 20% hexane/EtOAc (20 mL) and water (50 mL) . The organic layer was dried over sodium sulfate, and the solvent was removed with a rotary evaporator. The crude product was chromatographed on silica gel (30% EtOAc/hexane) , to afford 74 mg (50%) of the title compound.
Using substantially the same procedure as in Example 51, the items of the following Examples 52 - 82 can similarly be prepared.
EXAMPLE 52 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino)acetyl] -amino] -3- (biphenylthio) -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- (syn-methoxyimino) acetic acid and (71?) -7-amino-3- (biphenylthio) -3-cephem-4-carboxylate, benzyhydryl ester.
EXAMPLE 53 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-hydroxyimino) acetyl] -amino] -3- (biphenylthio) -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- (syn-hydroxyimino) acetic acid and (71?) -7-amino-3- (biphenylthio) -3-cephem-4-carboxylate, benzyhydryl ester.
EXAMPLE 54 (71?) -7- t [2- (2-Amino-4-thiazolyl) -2- (syn-hydroxyimino)acetyl] -amino] -3- (2-iodophenylthio) -3-cephem-4-carboxylate,
4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- (syn-hydroxyimino) acetic acid and (71?) -7-amino-3- (2-iodophenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 55 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-hydroxyimino)acetyl] -amino] -3- [4- (ethoxycarbonyl)phenylthio) ] -
3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- (syn-hydroxyimino) acetic acid and (71?) -7-amino-3- [4- (ethoxycarbonyl)phenylthio] -3-cephem-4-car boxylate, 4-methoxybenzyl ester.
EXAMPLE 56 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- [ [ [syn- (tert-butoxy) carbo nyl-methoxyimino] acetyl] amino] -3- [4- (2-thienyl)phenylthio] -3
-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 was employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- [syn- (tert-butoxy) carbonylmethoxyi mino] acetic acid and
(71?) -7-amino-3- [4- (2-thienyl)phenylthio] -3-cephem- -carboxyl ate, 4-methoxybenzyl ester.
EXAMPLE 57 EXAMPLE 57
(71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- [ [ [syn- (tert-butoxy) carbo nylmethoxy-imino] acetyl] amino] -3- [4- (2-furyl)phenylthio] -3-c ephem-4-carboxylate,4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- [syn- ( tert-butoxy) carbonylmethoxyi mino] acetic acid and
(71?) -7-amino-3- [4- (2-furyl)phenylthio] -3-cephem-4-carboxylat e, 4-methoxybenzyl ester.
EXAMPLE 58 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) - acetyl] amino] -3- (2-iodophenylthio) -3-cephem-4-carboxylate,
4-methoxybenzyl ester The procedure of Example 51 is employed, but the reactants are
2- (2-amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) acetic acid and
(71?) -7-amino-3- (2-iodophenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 59
(71?) - [ (2-Thienylacetyl) amino] -3- [6- (chloromethyl)dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester The procedure of Example 51 is employed, but the reactants are 2-thienylacetyl chloride-pyridine and (71?) -7-amino-3- [6- (chloromethyl) dibenzofuran-3-ylthio] -3-cep hem-4-carboxylate, benzhydryl ester. EXAMPLE 60
(71?) - [ (2-Thienylacetyl)amino] -3- [6- (chloromethyl) dibenzofuran-1-ylthio] -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2-thienylacetyl chloride-pyridine and
(71?) -7-amino-3- [6- (chloromethyl) dibenzofuran-1-ylthio] -3-cep hem-4-carboxylate, benzhydryl ester.
EXAMPLE 60a (71?) - [ (2-Thienylacetyl) amino] -3- [6- (chloromethyl) dibenzofura n-1-ylthio] -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2-thienylacetyl chloride-pyridine and (71?) -7-amino-3- [6- (chloromethyl) dibenzofuran-1-ylthio] -3-cep hem-4-carboxylate, benzhydryl ester.
EXAMPLE 61
(71?)- [[ (2-Amino-4-thiazolyl) acetyl] amino] -3- (2-bromophenylth io) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are (2-amino-4-thiazolyl) acetic acid and (71?) -7-amino-3- (2-bromophenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 62
(71?) - [ [ (2-Amino-4-thiazolyl) acetyl] amino] -3- (2-iodophenylthi o) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are (2-amino-4-thiazolyl) acetic acid and (71?) -7-amino-3- (2-iodophenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 63
(71?) - [ (2-Fury1acetyl)amino] -3- [4- (2-oxazolylmethyl)phenylthi o] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are 2-furylacetic acid and
(71?) -7-amino-3- [4- (2-oxazolylmethyl)phenylthio) -3-cephem-4-c arboxylate, 4-methoxybenzyl ester.
EXAMPLE 63a (71?) - [ (2-Furylacetyl)amino] -3- [4- (2-imidazolylmethyl) phenylthio] -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2-furylacetic acid and
(71?) -7-amino-3- [4- (2-imidazolylmethyl)phenylthio) -3-cephem-4 -carboxylate, benzhydryl ester.
EXAMPLE 64 (71?) - [ (4-Bromophenylacetyl)amino] -3- [dibenzofuran-3-ylthio] -
3-cephem-4-carboxylate, benzyhydryl ester
The procedure of Example 51 is employed, but the reactants are 4-bromophenylacetyl chloride and (71?) -7-amino-3- [dibenzofuran-3-ylthio] -3-cephem-4-carboxylat e, benzhydryl ester.
EXAMPLE 65
(71?) - [ (2-Iodophenylacetyl)amino] -3- [2-bromophenylthio] -3-cep hem-4-carboxylate, benzyhydryl ester The procedure of Example 51 was employed, but the reactants are 2-iodophenylacetyl chloride and (71?) -7-amino-3- [2-bromophenylthio] -3-cephem-4-carboxylate, benzhydryl ester.
EXAMPLE 66
(71?) - [ [ (2-Iodophenylthio)acetyl] amino] -3- [2-iodophenylthio] - 3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are 2- (iodophenythio) acetic acid and (71?) -7-amino-3- [2-iodophenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 67 (71?) - [ [ (2-Iodophenylthio)acetyl] amino] -3- (biphenylthio) -3-ce phem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are 2- (iodophenythio) acetic acid and (71?) -7-amino-3- (biphenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 68
(71?) - [ [ (Trifluoromethylthio)acetyl] amino] -3- [4- (2-oxazolyl)p henyl-thio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are (trifluoromethylthio) acetic acid and (71?) -7-amino-3- [4- (2-oxazolyl)phenylthio] -3-cephem-4-carboxy late, 4-methoxybenzyl ester.
EXAMPLE 69 (71?) - [[ (Trifluoromethylthio)acetyl] amino] -3- [2-bromo-4- (etho xycarbonyl)phenylthio] -3-cephem-4-carboxylate,
4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are (trifluoromethylthio)acetic acid and (71?) -7-amino-3- [2-bromo-4- (ethoxycarbonyl)phenylthio] -3-ceph em-4-carboxylate, 4-methoxybenzyl ester.
EXAMPLE 70
(71?) -7- [ [2- (2-Furyl) -2- (syn-methoxyimino)acetyl] amino] -3- (2- bromophenylthio) -3-cephem-4-carboxylate, benzhydryl ester The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
(71?) -7-amino-3- (2-bromophenylthio) -3-cephem-4-carboxylate, benzhydryl ester. EXAMPLE 71
(71?) -7- [ [2- (2-Furyl) -2- (syn-methoxyimino) acetyl] amino] -3- (di benzo-furan-2-ylthio) -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
(71?) -7-amino-3- (dibenzofuran-2-ylthio) -3-cephem-4-carboxylat e, benzhydryl ester.
EXAMPLE 72
(71?) -7- [ [2- (2-Furyl) -2- (syn-methoxyimino) acetyl] amino] -3- [4- (1-methyl-2-imidazolyl)phenylthio] -3-cephem-4-carboxylate, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
(71?) -7-amino-3- [4- (l-methyl-2-imidazolyl) -3-cephem-4-carboxy late, benzhydryl ester.
EXAMPLE 73
(71?) -7- [ [2- (2-Furyl) -2- (methoxyimino) acetyl] amino] -3- [6- (hydroxy-methyl) dibenzofuran-3-ylthio] -3-cephem-4-carboxy late, benzhydryl ester
The procedure of Example 51 is employed, but the reactants are 2- (2-furyl) -2- (syn-methoxyimino) acetic acid and
(71?) -7-amino-3- [6- (hydroxymethyl) dibenzofuran-3 -ylthio] -3-ce phem-4-carboxylate, benzhydryl ester. EXAMPLE 74
(71?) -7- [ (D-Phenylglycyl)amino] -3- (dibenzofuran-1-ylthio) -cep h-3-em-4-carboxylate, benzhydryl ester
This is prepared by the reaction of D-phenylglycyl chloride hydrochloride and (7R) -7-amino-3- (dibenzofuran-1-ylthio) -3-cephem-4-carboxylat e, benzhydryl ester in the presence of tetrahydrofuran and aqueous sodium bicarbonate.
EXAMPLE 75 (71?) -7- [ (D-Phenylglycyl) amino] -3- (dibenzofuran-2-ylthio) -cep h-3-em-4-carboxylate, benzhydryl ester
This is prepared by the reaction of D-phenylglycyl chloride hydrochloride and
(7R) -7-amino-3- (dibenzofuran-2-ylthio) -3-cephem-4-carboxylat e, benzhydryl ester in the presence of tetrahydrofuran and aqueous sodium bicarbonate.
EXAMPLE 76 (71?) -7- [ (D-Phenylglycyl) amino] -3- (dibenzofuran-3-ylthio) -cep h-3-em-4-carboxylate, benzhydryl ester
This is prepared by the reaction of D-phenylglycyl chloride hydrochloride and
(7R) -7-amino-3- (dibenzofuran-3-ylthio) -3-cephem-4-carboxylat e, benzhydryl ester in the presence of tetrahydrofuran and aqueous sodium bicarbonate. EXAMPLE 77
(71?) -7- [(D- (2-Formyloxy)phenylacetyl) amino] -3- [4- (2-furyl) phenyl-thio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are D- (2-formyloxy)phenylacetic acid and (71?) -7-amino-3- [4- (2-furyl)phenylthio) -3-cephem-4-carboxylat e, 4-methoxybenzyl ester.
EXAMPLE 78 (71?) -7- [ (Cyanoacetyl)amino] -3- [3- (imidazolylmethyl) phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
The procedure of Example 51 is employed, but the reactants are cyanoacetic acid and
(71?) -7-amino-3- [3- (imidazolylmethyl)phenylthio] -3-cephem-4-c arboxylate, 4-methoxybenzyl ester.
EXAMPLE 79 (71?) -7- [ (Cyanoacetyl)amino] -3- [3-dibenzothien-l-ylthio] -3-ce phem-4-carboxylate, 4-methoxybenzyl ester This is prepared by a procedure similar to that for Example 51, but the reactants are cyanoacetic acid and
(71?) -7-amino-3- [3-dibenzothien-l-ylthio] -3-cephem-4-carboxyl ate, 4-methoxybenzyl ester.
EXAMPLE 80 (71?) -7- [ (Cyanoacetyl)amino] -3- [4- (trifluoromethyl) phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This is prepared by a procedure similar to that for Example 51, but the reactants are cyanoacetic acid and (71?) -7-amino-3- [4- (trifluoromethyl)phenylthio] -3-cephem-4-ca rboxylate, 4-methoxybenzyl ester.
EXAMPLE 81 (71?) -7- [ (a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl) amino] -3- [2-iodo-4- (cyanamido)phenylthio] -3-ceph em-4-carboxylate, 4-methoxybenzyl ester
This is prepared by the reaction of a- [D-4-ethyl-2, 3-dioxo-l-piperazine-carboxamido] -a-phenylace tyl chloride and
(71?) -7-amino-3- [2-iodo-4- (cyanamido)phenylthio] -3-cephem-4-c arboxylate, 4-methoxybenzyl ester in the presence of tetrahydrofuran and aqueous sodium bicarbonate.
EXAMPLE 82 (71?) -7- [(a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl)amino] -3- [dibenzofuran-3-ylthio] -3-cephem-4-carb oxylate, 4-methoxybenzyl ester
This is prepared by the reaction of a- [D-4-ethyl-2, 3-dioxo-l-piperazine-carboxamido] -a-phenylace tyl chloride and (71?) -7-amino-3- [dibenzofuran-3-ylthio] -3-cephem-4-carboxylat e, 4-methoxybenzyl ester in the presence of tetrahydrofuran and aqueous sodium bicarbonate. EXAMPLE 83
(71?) -7- [ (phenylacetyl)amino] -3- [6- (chloromethyl) dibenzofuran-3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester
This is prepared by the reaction of (71?) -7- [ (phenylacetyl) amino] -3- [6- (hydroxymethyl) dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
Under substantially the same procedure as in Example 83, the items of the following Examples 84 - 88 are similarly prepared.
EXAMPLE 84 (71?) -7- [ (phenylacetyl)amino] -3- [7- (chloromethyl)dibenzofuran -3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This is prepared by the reaction of (71?) -7- [ (phenylacetyl) amino] -3- [7- (hydroxymethyl) dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
EXAMPLE 85 (71?) -7- [ (2-Furylacetyl)amino] -3- [7- (chloromethyl) dibenzofuran-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This is prepared by the reaction of (71?) -7- [ (2-furylacetyl) amino] -3- [7- (hydroxy-methyl) dibenzofu ran-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
EXAMPLE 86 (71?) -7- [2- (2-Thienylacetyl) -2- (syn-methoxyimino) acetyl] amino ] -3- [4- [41 - (chloromethyl)phenyl]phenylthio] -3-cephem-4-carbo xylate,4-methoxybenzyl ester
This is prepared by the reaction of (71?) -7- [ (2-thienylacetyl) amino] -3- [4- [4 ' - (hydroxymethyl)phen yl]phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
EXAMPLE 87 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (sy -2-fluoroethoxyimino) -acetyl] amino] -3- [4- (chloromethyl) phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This is prepared by the reaction of (71?) -7- [ [2- (2-amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) acetyl) amino] -3- [4- (hydroxymethyl)phenylthio] -3-cephem-4-car boxylate, 4-methoxybenzyl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
EXAMPLE 88 (71?) -7- [ (a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl)amino] -3- [4- (chloromethyl)phenylthio] -3-cephem-4 -carboxylate, 4-methoxybenzyl ester
This is prepared by the reaction of 7- [(a- [D-4-ethyl-2, 3-dioxo-l-piperazine-carboxamido] -a-pheny lacetyl) amino] -3- [4- (hydroxymethyl)phenylthio] -3-cephem-4-ca rboxylate, 4-methoxybenzyl ester with 1.1 equivalents of thionyl chloride and pyridine in methylene chloride at 0°C.
EXAMPLE 89 (71?) -7- [ (phenylacetyl)amino] -3- [6- (pyridin-1-iummethyl)diben zofuran-3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester, chloride salt
This is made by the reaction of (71?) -7- [ (phenylacetyl) amino] -3- [6- (chloromethyl) dibenzofuran -3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 90 (71?) -7- [ (phenylacetyl)amino] -3- [6- (N-methylimidazol-1-iummet hyl) -dibenzofuran-3-ylthio] -ceph-3-em-4-carboxylate, benzhydryl ester, chloride salt
This is made by the reaction of (71?) -7- [ (phenylacetyl) amino] -3- [6- (chloromethyl) dibenzofuran -3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester with 4.0 equivalents of 1-methylimidazole in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 91
(71?) -7- [ (phenylacetyl) amino] -3- [7- (pyridin-1-iummethyl) -dibe nzofuran-3-ylthio] -ceph-3-em-4-carboxylate, benzhydryl ester, chloride salt
This is made by the reaction of (71?) -7- [ (phenylacetyl) amino] -3- [7- (chloromethyl) dibenzofuran -3-ylthio] -3-cephem-4-carboxylate, benzhydryl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 92
(71?) -7- [ (2-Furylacetyl)amino] -3- [7- (N-methylimidazol-1-iumme thyl) -dibenzofuran-3-ylthio] -ceph-3-em-4-carboxylate,
4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ (2-furylacetyl) amino] -3- [7- (chloromethyl) dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 93
(71?) -7- [ (2-Furylacetyl) amino] -3- [6- (pyridin-1-iummethyl) dibe nzo-furan-2-ylthio] -ceph-3-em-4-carboxylate, 4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ (2-furylacetyl) amino] -3- [6- (chloromethyl) dibenzofura n-3-ylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether. EXAMPLE 94
(71?) -7- [ [2- (2-Thienyl) -2- (syn-methoxyimino) acetyl] amino] -3- [
4- (N-pyridin-l-iummethyl)phenylthio] -ceph-3-em-4-carboxylate
, 4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ [2- (2-thienylacetyl) -2- (syn-methoxy-imino) acetyl] ami no] -3- [4- (chloromethyl)phenylthio] -ceph-3-em-4-carboxylate, 4-methoxybenzyl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 95
(71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) acetyl] -amino] -3- [4- (N-methylimidazol-1-iummethyl)phenylthio
] -ceph-3-em-4-carboxylate, 4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ [2- (2-amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) acetyl] amino] -3- [4- (chloromethyl)phenylthio] -ceph-3-em-4-car boxylate, 4-methoxybenzyl ester with 4.0 equivalents of 1-methylimidazole in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 96 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn- (tert-butoxy)carbony 1-methoxyimino)acetyl] -amino] -3- [4-
(N-methylimidazol-1-iummethyl)phenylthio] -ceph-3-em-4-carbox ylate, 4-methoxybenzyl ester, chloride salt This is made by the reaction of (71?) -7- [ [2- (2-amino-4-thiazolyl) -2- ( syn- ( tert-butoxy) carbony Imethoxyimino) acetyl] amino] -3- [4- (chloromethyl) phenylthio] -c eph-3-em-4-carboxylate, 4-methoxybenzyl ester with 4.0 equivalents of 1-methylimidazole in acetonitrile. Desired material was obtained by precipitation with diethyl ether.
EXAMPLE 97
(71?) -7- [ (a- [D-4-Ethyl-2, 3-dioxo-l-piperazinecarboxamido] -a-p henylacetyl) amino] -3- [4- (pyridin-l-iummethyl)phenylthio] -3-c ephem-4-carboxylate, 4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ (a- [D-4-ethyl-2 , 3-dioxo-l-piperazine-carboxamido] -a- phenylacetyl) amino] -3- [4- (chloromethyl)phenylthio] -3-cephem- 4-carboxylate, 4-methoxybenzyl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 98
(71?) -7- [ (2,4-Dibromophenylacetyl) amino] -3- [4 ' -
(pyridin-1-ium- ethyl) iphenylthio] -ceph-3-em-4-carboxylate,
4-methoxy-benzyl ester, chloride salt This is made by the reaction of
(71?) -7- [ (2 , 4-dibromophenylacetyl) amino] -3- [4 ' - (chloromethyl) biphenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether. EXAMPLE 99
(71?) -7- [ [ (2,4-Dichlorophenylthio)acetyl] amino] -3- [2-bromo-4-
(N-methylimidazol-1-iummethyl)phenylthio] -ceph-3-em-4-carbox ylate, 4-methoxybenzyl ester, chloride salt
This is made by the reaction of (71?) -7- [ [ (2, 4-dichlorophenylthio) acetyl] amino] -3- [2-bromo-4- (chloromethyl) phenylthio] -3-cephem-4-carboxylate, 4-methoxy-benzyl ester with 4.0 equivalents of 1-methylimidazole in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 100
(71?) -7- [ [ (2,4-Dichlorophenylthio)acetyl] amino] -3- [2-bromo-4-
(pyridin-1-iummethyl)phenylthio] -ceph-3-em-4-carboxylate,
4-methoxybenzyl ester, chloride salt This is made by the reaction of
(71?) -7- [ [ (2, 4-dichlorophenylthio) acetyl] amino] -3- [2-bromo-4- (chloromethyl) phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester with 4.0 equivalents of pyridine in acetonitrile. Desired material is obtained by precipitation with diethyl ether.
EXAMPLE 101 (71?) -7- [ (phenylacetyl)amino] -3- (4-biphenylthio) -3-cephem-4-c arboxylic acid A solution of
(71?) -7- [ (phenylacetyl) amino] -3- (4-biphenylthio) -3-cephem-4-c arboxylate, benzhydryl ester (450 mg, 0.670 mmol) and anisole (0.4 mL) in dichloromethane (3 mL) at 0° is treated with trifluoroacetic acid (4 mL) . After lh, the volatiles are removed with a rotary evaporator. The residue was subjected to column chromatography on silica gel (dichloromethane, then 10% AcOH/dichloromethane) , affording 232 mg (69%) of the title compound. XH NMR (acetone-d6) d 3.37 (d, IH, J=18) , 3.63 (d, IH, J=18) , 3.84 (d, IH, J=16) , 3.92 (d, IH, J=16), 5.25 (d, IH, J=5), 5.93 (dd, IH, J=5,9), 7.2-7.8 (m, 14H) , 8.15 (d, IH, J=9) ; IR (KBr) 1513, 1619, 1713, 1784, 3377cm"1.
EXAMPLE 102 (71?) -7- [ (phenylacetyl)amino] -3- (4-bromophenylthio) -3-cephem-
4-carboxylic acid
To a solution of (71?) -7- [ (phenylacetyl) amino] -3- (4-bromophenylthio) -3-cephem- 4-carboxylate, benzhydryl ester (80 mg, 0.12 mmol) in phenol (230 mg) at 45° was added trifluoroacetic acid (9 μL) . After 1.5 h, the volatiles are removed with a rotary evaporator, and the residue is triturated with isopropanol/hexane. The resulting white solid was washed with ether and dried in vacuo, affording 48 mg (79%) of the title compound. XH NMR (acetone-d6) d 3.67 (d, IH, J=16) , 3.75 (d, IH, J=16) , 3.64 (d, IH, J=18) , 3.67 (d, IH, J=18) , 5.25 (d, IH, J=5) , 5.92 (d, IH, J=5) , 7.2-7.7 (m, 9H) ; IR (KBr) 1347, 1386, 1606, 1756, 3276 cm"1. EXAMPLE 103
(71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino) acetyl] amino] -3- (4-bromophenylthio) -3-cephem-4-carboxylic acid
To a mixture of (71?) -7- [ [ [2- (2-amino-4-thiazolyl) -2- (syn-methoxyimino) ] acety 1] -amino] -3- (4-bromophenylthio) -3-cephem-4-carboxylate, benzhydryl ester (71 mg, 0.096 mmol) and anisole (0.2 mL) at 0° was added trifluoroacetic acid. After 1 h, the volatiles are removed with a rotary evaporator. The crude product was subjected to reverse-phase chromatography on C18 adsorbent (acetonitrile/ water 3:2) , affording 11.6 mg (21%) of the title compound.
Under substantially the same procedures as in Examples 101 - 103, the items of the following Examples 104 - 141 can similarly be obtained.
EXAMPLE 104
(71?) -7- [ [ (2-Amino-4-thiazolyl)acetyl] amino] -3- (2-bromophenyl thio) -3-cephem-4-carboxylic acid
EXAMPLE 105
(71?) -7- [ [ (2-Amino-4-thiazolyl)acetyl] amino] -3- (2-iodophenylt hio) -3-cephem-4-carboxylic acid
EXAMPLE 106 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-hydroxyimino) acetyl] amino] -3- [4- (ethoxycarbonyl)phenylthio] -3-cephem-4-ca rboxylic acid
EXAMPLE 107 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino)acetyl] amino] -3- (2-iodophenylthio) cephem-4-carboxylic acid
EXAMPLE 108 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-2-fluoroethoxyimino) acetyl] mino] -3- (2-iodophenylthio) -3-ceρhem-4-carboxylic acid
EXAMPLE 109 (71?) -7- [ [2- (2-Amino-4-thiazolyl) -2- (syn-carboxymethoxyimino) acetyl] amino] -3- [4- (2thienyl)phenylthio] -3-cephem-4-carboxyl ic acid
EXAMPLE 110 (71?) -7- [ (a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a- ( hydroxy-phenyl)acetyl)amino] -3- [2-iodo-4- (cyanoamido)phenyl¬ thio] -3-cephem-4-carboxylic acid
EXAMPLE 111 (71?) -7- [(a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido] -a- (hydroxyphenyl)acetyl)amino] -3- [dibenzofuran-3-ylthio] -3-cep hem-4-carboxylic acid
EXAMPLE 112 (71?) -7- [ (2-Iodophenylthioacetyl) amino] -3- (2-iodophenylthio) - 3-cephem-4-carboxylic acid
EXAMPLE 113 (71?) -7- [ (2-Iodophenylthioacetyl)amino] -3- (biphenylthio) -3-ce phem-4-carboxylic acid
EXAMPLE 114 (71?) -7- [ (Cyanoacetyl) amino] -3- [3- (imidazolylmethyl) phenylthio] -3-cephem-4-carboxylic acid
EXAMPLE 115 (71?) -7- [ (Cyanoacetyl) alnino] -3- (dibenzothienyl-1-ylthio) -3-c ephem-4-carboxylic acid
EXAMPLE 116
(71?) -7- [ (Cyanoacetyl) amino] -3- [4- (trifluoromethyl) phenylthio) -3-cephem-4-carboxylic acid
EXAMPLE 117 (71?) -7- [ (2-Thienylacetyl)amino] -3- [6- (chloromethyl) dibenzofuran-3-ylthio] -3-cephem-4-carboxylic acid
EXAMPLE 118 (71?) -7- [ (2-Thienylacetyl) amino] -3- [dibenzofuran-3-ylthio] -3- cephem-4-carboxylic acid
EXAMPLE 119
(71?) -7- [ (2-Furylacetyl) amino] -3- [4- (2-oxazolylmethyl) phenylthio] -3-cephem-4-carboxylic acid EXAMPLE 120
(71?) -7- [ (2-Furylacetyl)amino] -3- [4- (1-imidazolymethyl) phenylthio] -3-cephem-4-carboxylic acid
EXAMPLE 121
(71?) -7- [ ( (Trifluoromethyl) thioacetyl)amino] -3- [4- (2-oxazolyl )phenylthio] -3-cephem-4-carboxylic acid
EXAMPLE 122 (71?) -7- [ ( (Trifluoromethyl) thioacetyl)amino] -3- [2-bromo-4- (et hoxy-carbonyl) henylthio] -3-cephem-4-carboxylic acid
EXAMPLE 123 (7JF?) -7- [ (D- (2-Formyloxy)phenylacetyl)amino] -3- [4- (2-furyl)ph enylthio] -3-cephem-4-carboxylic acid
EXAMPLE 124 (71?) -7- [ (phenylacetyl)amino] -3- (dibenzofuran-1-ylthio) -3-cep hem-4-carboxylic acid
EXAMPLE 125 (71?) -7- [ (phenylacetyl)amino] -3- (dibenzofuran-3-ylthio) -3-cep hem-4-carboxylic acid
EXAMPLE 126
(71?) -7- [ (phenylacetyl)amino] -3- (6-hydroxymethyldibenzofuran- 3-ylthio) -3-cephem-4-carboxylic acid
EXAMPLE 127 (71?) -7- [ (phenylacetyl) amino] -3- (7-hydroxymethyldibenzofuran- 3-ylthio) -3-cephem-4-carboxylic acid
EXAMPLE 128 (71?) -7- [ (D-Phenylglycyl)amino] -3- (dibenzofuran-1-ylthio) -3-c ephem-4-carboxylic acid
EXAMPLE 129 (71?) -7- [ (D-Phenylglycyl)amino] -3- (dibenzofuran-2-ylthio) -3-c ephem-4-carboxylic acid
EXAMPLE 130
(71?) -7-{ [2- (2-Furyl) -2- (syn-methoxyimino)acetyl] amino}-3- (di benzofuran-1-ylthio) -3-cephem-4-carboxylic acid
EXAMPLE 131
(71?) -7-{ [2- (2-Furyl) -2- (syn-methoxyimino)acetyl] amino}-3- (di benzofuran-2-ylthio) -3-cephem-4-carboxylic acid
EXAMPLE 132 (7JF?)-7-[(2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino) acetyl) amino] -3- (6-hydroxymethyldibenzofuran-2-ylthio) -3-cep hem-4-carboxylic acid
EXAMPLE 133 (71?) -7- [ ( (2-Amino-4-thiazolyl)acetyl)amino] -3 (6-hydroxymethy ldibenzo-furan-3-yl-thio) -3-cephem-4-carboxylic acid
EXAMPLE 134 (71?) -7-{ [2- (2-Furyl) -2- (methoxyimino) acetyl] amino}-3- (7-hydr oxymethyl-dibenzofuran-3-ylthio) -3-cephem-4-carboxylic acid
EXAMPLE 135
(71?) -7-{ [2- (2-Furyl) -2- (methoxyimino) acetyl] amino}-3-
(7-hydroxymethyl-dibenzofuran-1-ylthio) -3-cephem-4-carboxyli c acid
EXAMPLE 136
(71?) -7- [ (phenylacetyl)amino] -3- [6- (pyridin-immethyl-1) dibenzofuran-3-ylthio] -3-cephem-4-carboxylate, inner salt
EXAMPLE 137
(71?) -7- [ (phenylacetyl)amino] -3- [6- (N-methylimidazol-1-iummet hyl)dibenzofuran-3-ylthio] -ceph-3-em-4-carboxylic acid, inner salt
EXAMPLE 138 (71?) -7- [ (2-Thienylacetyl)amino] -3- [6- (N-methylimidazol-1-ium methyl) dibenzofuran-3-ylthio] -ceph-3-em-4-carboxylic acid, inner salt
EXAMPLE 139
(71?) -7- [ (2-Thienylacetyl)amino] -3- [6- (pyridiniummethyl)diben zofuran-3-ylthio] ceph-3-em-4-carboxylic acid, inner salt EXAMPLE 140
(71?) -7- [ (2-Thienylacetyl)amino] -3- [6- (pyridiniummethyl)diben zofuran-2-ylthio] -ceph-3-em-4-carboxylic acid, inner salt
EXAMPLE 141 (71?) -7- [(2- (2-Amino-4-thiazolyl) -2- (syn-hydroxyimino)acetyl] amino-3- (2-iodophenylthio) -3-cephem-4-carboxylic acid
EXAMPLE 142 (71?) -7- [ (phenylacetyl)amino] -3- [4- [ (3-hydroxymethyl)phenyl] - phenylthio] -3-cephem-4-carboxylic acid, sodium salt (Pd coupling)
To a stirred solution of (71?) -7- [ (phenylacetyl) amino] -3- (4-iodophenyl-thio) -3-cephem- 4-carboxylic acid (25.0 mg, 0.045 mmol) , 3-hydroxymethyl-1-trimethylstannylbenzene (16.3 mg, 0.060 mmol), tri (2-furyl)phosphine (1.1 mg) , sodium bicarbonate (4.2 mg, 0.045 mmol) and 2, 6-di-t-butyl-4-methylphenol (2 mg) in N-methylpyrrolidinone (0.3 mL) was added tris (dibenzylideneacetone) dipalladium(0) (4.5 mg, 0.0049 mmol) . The mixture was heated at 65° for 2 h, at which time the reaction was quenched by addition of 1% aqueous sodium bicarbonate solution (10 mL) . The resulting solution was subjected to reverse phase chromatography (C18 adsorbent, 30% acetonitrile/water) , affording 9.2 mg (37%) of the title compound. Η NMR (acetone-d6) d 3.35 (d, IH, J=16) , 3.38 (d, IH, J=16) , 3.64 (d, IH, J=18) , 3.67 (d, IH, J=18) , 5.23 (d, IH, J=5) , 5.83 (dd, IH, J=5,9), 7.2-7.8 (m, 13H) , 8.15 (d, IH, J=9) ; IR (KBr) 1387, 1610, 1654, 1762, 3264cm-1. Substantially following the method described in detail hereinabove in Example 142, the compounds of this invention listed below in Examples 143-156 can be prepared.
EXAMPLE 143 (71?) -7- [ (Thienylacetyl) amino] -3- [4- (2-imidazolyl) phenylthio] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 144 (71?) -7- [ (2,4-Dichlorophenylthio)acetylamino] -3- [4- [(1-methyl -imidazol-4-yl)phenylthio] ] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 145 (71?) -7- [ (2,4-Difluorophenylthio) acetylamino] -3- [4- [ (l-methyl-imidazol-4-yl)phenylthio] ] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 146 (71?) -7- [ [2- (2-Furyl) -2- (syn-methoxyimino) acetyl] amino] -3- [4- [ (l-methylimidazol-4-yl)phenylthio] ] -3-cephem-4-carboxyli c acid, sodium salt
EXAMPLE 147 (71?) -7- [a- [D-4-Ethyl-2,3-dioxo-l-piperazinecarboxamido]pheny lacetylamino] -3- [4- [ (1-methylimidazol-4-yl)phenylthio] ] -3-ce phem-4-carboxylic acid, sodium salt
EXAMPLE 148 (71?) -7- [D- [ (2-Formyloxy)phenylacetyl] amino] -3- [4- [(1-methyl- imidazol-4-yl)phenylthio] ] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 149 (71?) -7- [ (2- (2-Amino-4-thiazolyl) -2- (syn-methoxyimino) acetyl] amino-3- [4- [ (1-methylimidazol-4-yl)phenylthio] ] -3-cep hem-4-carboxylic acid, sodium salt
EXAMPLE 150 (71?) -7- [ (2- (2-Amino-4-thiazolyl) -2- (syn-cyclopentoxyimino)ac etyl] amino-3- [4- [ (1-methylimidazol-4-yl) henylthio] ] -3-σephe m-4-carboxylic acid, sodium salt
EXAMPLE 151 (71?) -7- [ (2- (2-Amino-4-thiazolyl) -2- (syn-cyclobutoxyimino)ace tyl] mino-3- [4- [ (l-methyl-imidazol-4-yl)phenylthio] ] -3-cephe m-4-carboxylic acid, sodium salt
EXAMPLE 152 (71?) -7- [ (D- (2-Formyloxy) henylacetyl)amino] -3- [4- (2-furyl)ph enylthio] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 153
(71?) -7- [ (4-Chlorophenylacetyl)amino] -3- [2- (2-furyl) phenylthio] -3-cephem-4-carboxylic acid, sodium salt EXAMPLE 154
(71?) -7- [ [ (4-Methoxyphenylthio)acetyl] amino] -3- [4-
(trifluoro-methoxy) -2- (phenyl)phenylthio] -3-cephem-4-carboxy lie acid, sodium salt
5 EXAMPLE 155
(71?) -7- [ (2-Thienylacetyl)amino] -3- [2- (3,4-dicyanophenyl)phen ylthio] -3-cephem-4-carboxylic acid, sodium salt
EXAMPLE 156 10 (71?) -7- [ (phenylacetyl)amino] -3- [2- (2-furyl)phenylthio] -3-cep hem-4-carboxylic acid, sodium salt
EXAMPLE 157 (71?) -7- [ (phenylacetyl) -amino] -3- (2-hydroxymethylphenylthio) -3 15 -cephem-4-carboxylate, benzhydryl ester
A solution of sodium 2-hydroxymethylbenzenethiolate was prepared by addition of a solution of sodium bis (trimethylsilyl) amide (1.0 M in THF, 1.86 mL, 1.86 mmol) to a solution of 2-mercaptobenzyl 20 alcohol (276 mg, 1.96 mmol) in DMF (8 mL) . After 10 min, the resulting yellow-orange solution was concentrated with a rotary evaporatory in order to remove the THF. This solution was added dropwise to a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy) ' 25 -3-cephem-4-carboxylate, benzhydryl ester (1.00 g, 1.58 mmol) in dry DMF (15 mL) at -45 °C under argon. After 20 min at -45 °C, the mixture was quenched by addition of saturated aqueous ammonium chloride solution, and was poured into stirring water (200 mL) . After 10 min, the resulting white precipitate was filtered and dried, affording 0.93 g (95%) of the title compound. lH NMR (CDC13) d 3.03 (d, 1, J=18) , 3.09 (d, 1, J=18), 3.62 (d, 1, J=16) , 3.66 (d, 1, J=16) , 4.72 (d, 1, J=14) , 4.76 (d, 1 , J---14) , 4.97 (d, 1, J=4) , 5.75 (dd, 1, J=4,9) , 6.05 (d, 1, J=9) , 6.98 (s, 1) , 7.2-7.6 (m, 19) .
EXAMPLE 158
(71?) -7- [ (phenylacetyl)amino] -3- (2-bromomethylphenylthio) -3-c ephem-4-carboxylate, benzhydryl ester To a stirring solution of triphenylphosphine (90.5 mg, 0.34 mmol) in THF (5 mL) at room temperature under argon was added carbon tetrabromide (80 mg, 0.24 mmol) . After 20 min, to this cloudy white mixture was added a solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-hydroxymethylphenylthio) -3 -cephem-4-carboxylate, benzhydryl ester (124 mg, 0.20 mmol) in THF (3 mL) . After stirring overnight at room temperature, the mixture was concentrated with a rotary evaporator, and the residue was subjected to flash chromatography on silica gel (5 g, 2% methanol/dichloromethane) , affording 66 mg (48%) of the title compound. lH NMR (CDC13) d 3.15 (d, 1, J=18) , 3.23 (d, 1, J=18) , 3.60 (d, 1, J=16) , 3.64 (d, 1, J=16) , 4.61 (s, 2) , 5.00 (d, 1, J=4)', 5.78 (dd, 1, J=4,9) , 6.17 (d, 1, J=9) , 6.99 (s, 1) , 7.2-7.6 (m, 19) . EXAMPLE 159 (71?) -7- [ (phenylacetyl)amino] -3- (2-isothiouroniummethylphenyl thio) -3-cephem-4-carboxylate, benzhydryl ester, bromide salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-bromomethylphenylthio) -3-c 5 ephem-4-carboxylate, benzhydryl ester (66 mg, 0.096 mmol) in acetonitrile (1 mL) was added thiourea (6.0 mg, 0.079 mmol.) . After 1.5 h, the mixture was concentrated to a yellow oil and subjected to flash chromatography on silica gel (10% methanol/dichloromethane) , affording 30 mg (50%) of the 10 title compound. Η NMR (CDC13/CD30D) d 2.80 (d, 1, J=18) , 3.31 (d, 1, J=18) , 3.63 (s, 2) , 4.46 (s, 2), 5.05 (d, 1, J=4) , 5.57 (d, 1, J=4) , 6.91 (s, 1) , 7.1-7.5 (m, 19) .
EXAMPLE 160 15 (71?) -7- [ (phenylacetyl)amino] -3- (2-isothiouroniummethylphenyl thio) -3-cephem-4-carboxylic acid, trifluoroacetate salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-isothiouroniummethylphenyl thio) -3-cephem-4-carboxylate, benzhydryl ester, bromide salt 20 (30 mg) in dichloromethane (0.5 mL) at 0 °C was added a solution of anisole (0.2 mL) in trifluoroacetic acid (0.4 mL) . After 1 h at 0 °C, the mixture was concentrated with a rotary evaporator, and the residue was triturated with ether, filtered and dried to afford 11.8 mg of the title -25 compound. lH NMR (CDC13/CD30D) d 3.12 (d, 1, J=18) , 3.21 (d, 1, J=18) , 3.58 (d, 1, J=16) , 3.62 (d, 1, J=16) , 4.60 (d, 1, J=12) , 4.71 (d, 1, J=12) , 5.01 (d, 1, J=4) , 5.58 (d, 1, J=4) , 6.89 (d, 1, J=10) , 7.2-7.5 (m, 7) 7.58 (d, 1, J=10) . EXAMPLE 161
(7JF?) -7- [ (phenylacetyl)amino] -3- (2-hydroxymethylphenylthio) -3
-cephem-4-carboxylate, 4-methoxybenzyl ester
To a mixture of sodium methoxide (969 mg, 17.9 mmol) in dry THF (50 L) at 0 °C was added dropwise a solution of 2-mercaptobenzyl alcohol (2.51 g, 17.9 mmol) in dry THF (25 mL) . The resulting yellow solution was stirred at 0 °C for 30 min, after which the solvent was removed in vacuo . To the resulting thiolate salt was added (71?) -7- [ (phenylacetyl) amino] -3-trifluoromethanesulfonyloxy-3 -cephem-4-carboxylate, 4-methoxybenzyl ester (10.0 g, 17.0 mmol) , and this solid mixture was cooled to -40 °C. Dry DMF (100 mL) at -45 °C was introduced via cannula, and the resulting mixture was stirred at -45 ΛC, forming a yellow solution. After 30 min, the temperature was allowed to rise over 30 min to -10 °C, and the mixture was poured into ice water (1 L) . After stirring for 15 min, the mixture was filtered, and the precipitate was washed thoroughly with water and air-dried. Further washing with ether afforded the title compound (6.90 g, 71%) as an off-white solid. 1H NMR (CDC13) d 2.43 (br s, 1) , 3.06 (d, 1, J=18) , 3.15 (d, 1, J=18) , 3.60 (d, 1, J=16) , 3.63 (d, 1, J=16) , 3.81 (s, 3) , 4.75 (d, 1, J=14) , 4.78 (d, 1, J=14) , 4.93 (d, 1, J=4) , 5.24 (s, 2) , 5.70 (dd, 1, J=4,9) , 6.32 (d, 1, J=9) , 6.88 (d, 2, J=10) , 7.2-7.6 (m, 11) .
EXAMPLE 162
(71?) -7- [ (phenylacetyl)amino] -3- (2-bromomethylphenylthio) -3-c ephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for example 158, but
(71?) -7- [ (phenylacetyl) amino] -3- (2-hydroxymethylphenylthio) -3
-cephem-4-carboxylate, 4-methoxybenzyl ester was used.
EXAMPLE 163 (71?) -7-amino-3- (2-bromomethylphenylthio) -3-cephem-4-carboxyl ate, 4-methoxybenzyl ester
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-bromomethylphenylthio) -3-c ephem-4-carboxylate, 4-methoxybenzyl ester (1.24 g, 1.98 mmol) and pyridine (0.320 mL, 3.96 mmol) in dichloromethane (7 mL) at -10 °C was added slowly a solution of phosphorus pentachloride in dichloromethane (7.32 mL, 3.17 mmol) . After 2.5 h at -10 °C, the mixture was cooled to -40 °C, and isobutanol (1.82 mL, 19.8 mmol) was added. The mixture was allowed to warm to -10 °C, was stirred for Ih, and was partitioned between water and dichloromethane. The organic phase was washed with 10% aqueous HCl and saturated aqueous sodium bicarbonate, dried (sodium sulfate) , and concentrated with a rotary evaporator. the resulding beige solid was triturated with ether and dried, affording 504 mg (48%) of the title compound. lH NMR (CD30D) d 3.26 (d, 1, J=18) , 3.42 (d, 1, J=18) , 3.58 (s, 3) , 4.71 (d, 1, J=8) , 4.77 (d, 1, J=8) , 4.82 (s, 2) , 5.01 (d, 1, J=4) , 5.28 (d, 1, J=4) , 6.86 (d, 2, J=10) , 7.33 (d, 2, J=10) , 7.3-7.6 (m, 4) . EXAMPLE 164
(71?) -7- [ (4-bromomethylphenylacetyl)amino] -3- (2-bromomethylph enylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
To a stirring suspension of (71?) -7-amino-3- (2-bromomethylphenylthio) -3-cephem-4-carboxyl ate, 4-methoxybenzyl ester (200 mg, 0.374 mmol) and
4-bromomethylphenylacetic acid (103 mg, 0.449 mmol) in THF (2 mL) at -40 °C was added phosphorus oxychloride (41.7 μL, 0.449 mmol) followed be diisopropylethylamine (0.26 mL, 1.50 mmol) . The mixture was allowed to stir overnight at -10 °C. Water was added, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated aqueous sodium bicarbonate, 1 M phosphoric acid solution, and brine, dried (sodium sulfate) , filtered, and concentrated with a rotary evaporator. The crude product was purified by column chromatography on silica gel (0.5% methanol / dichloromethane) , affording 168 mg (61%) of the title compound. lH NMR (CDC13) d 3.15 (d, 1, J=18) , 3.27 (d, 1, J=18) , 3.61 (S, 2) , 3.82 (s, 3) , 4.46 (s, 2) , 4.65 (d, 1, J=8) , 4.68 (d, 1, J=8) , 4.95 (d, 1, J=4) , 5.26 (s, 2) , 5.74 (dd, 1, J=4, 9) , 6.20 (d, 1, J=9) , 6.86 (d, 2, J=10) , 7.2-7.5 (m, 10) .
EXAMPLE 165 (71?) -7- [ (4-isothiouroniummethylphenylacetyl)amino] -3- (2-isot hiouroniummethylphenylthio) -3-cephem-4-carboxylate,
4-methoxybenzyl ester, dibromide salt
To a stirring solution of (71?) -7- [ (4-bromomethylphenylacetyl) amino] -3- (2-bromomethylph enylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (168 mg, 0.229 mmol) in acetonitrile (5 mL) was added thiourea (34.9 mg, 0.458 mmol) . After stirring overnight at room temperature, the solvent was removed with a rotary evaporator, affording 203 mg (100%) of the title compound as a white-yellow powdery solid. "-H NMR (CD30D) d 4.02 (d, 1, J=18) , 4.31 (s, 2) , 4.36 (d, 1, J=18) , 4.54 (s, 3) , 5.25 (s, 2) , 5.40 (d, 1, J=10) , 5.43 (d, 1, J=10) , 5.95 (d, 1, J=12) , 5.99 (d, 1, J=12) , 6.02 (d, 1, J=4) , 6.50 (dd, 1, J=4, 9) , 7.65 (d, 2, J=10) , 8.0-8.2 (m, 10) , 8.39 (d, 1, J=9) , 9.97 (m, 8) .
EXAMPLE 166
(71?) -7- [ (4-isothiouroniummethylphenylacetyl)amino] -3- (2-isot hiouroniummethylphenylthio) -3-cephem-4-carboxylic acid, trifluoroacetate salt To a stirring suspension of
(71?) -7- [ (4-isothiouroniummethylphenylacetyl) amino] -3- (2-isot hiouroniummethylphenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester, dibromide salt (203 mg, 0.229 mmol) and anisole (0.1 mL) in dichloromethane (1 mL) at 0 °C was added trifluoroacetic acid (1 mL) . A solution formed, the reaction was stirred for 30 min, and the solvent was removed with a rotary evaporator. The residue was dissolved in water (150 mL, with sonication) , washed with diethyl ether, and filtered. The filtrate was lyophilized to afford 205 mg of the title compound. lH NMR (D20) d 3.20 (d, 1, J=18) , 3.38 (d, 1, J=18) , 3.61 (s, 2) , 4.40 (s, 2) , 4.61 (d, 1, J=8) , 4.64 (d, 1, J=8) , 5.15 (d, 1, J=4) , 5.59 (d, 1, J=4) , 7.2-7.6 (m, 8) .
SSSD/14615. vOl EXAMPLE 167
(71?) -7- [ (phenylacetyl)amino] -3- (2-phenylpyrid-3-ylthio) -3-ce phem-4-carboxylate, benzhydryl ester
A mixture of 2-bromo-3-hydroxypyridine (2.20 g, 12.6 mmol) , phenylboronic acid (2.00 g, 16.3 mmol) and palladium (tetrakis) triphenylphosphine (250 mg) in toluene
(30 mL) , ethanol (15 mL) and 5% aqueous sodium hydroxide (15 mL) was degassed with a stream of nitrogen and stirred vigorously at 100 °C overnight. The mixture was diluted with water (50 mL) and ethyl acetate (100 mL) , and the organic layer was dried over sodium sulfate and concentrated, affording a yellow solid. This material was suspended in water (100 mL) , a few mL of IN HCl was added, and the resulting solution was extracted with ethyl acetate. The aqeuous layer was neutralized with saturated aqueous sodium bicarbonate, and again was extracted with ethyl acetate. The combined organic extracts were dried with sodium sulfate and concentrated to an oily solid, which was triturated with 2:1 hexane/ethylacetate, affording 0.98 g of 3-hydroxy-2-phenylpyridine. To a stirring solution of
3-hydroxy-2-phenylpyridine (425 mg, 2.48 mmol) in DMF (8 mL) at room temperature was added a solution of sodium bis (trimethylsilyl) amide in DMF (2 M, 1.36 mL, 2.72 mmol) . After a few minutes, to this mixture was added a solution of N,N-dimethylthiocarbamoyl chloride (338 mg, 2.72 mmol) in DMF (2 mL) . After stirring overnight, the mixture was partitioned between water and ethyl acetate, and the organic phase was concentrated and subjected to flash chromatography on silica gel, affording 536 mg of * 3-N,N-dimethylthiocarbamoyl-2-phenylpyridine. Η NMR (CDC13) d 3.23 (s, 3) , 3.40 (s, 3) , 7.3-7.7 (m, 7) , 8.60 (d, 1, J=6) . This material was heated in a sealed vial at 250-255 °C for 1.5 h. The product was dissolved in ethanol (10 mL) , 10% aqueous potassium hydroxide was added (4 mL) , and the 5 resulting mixture was heated under nitrogen at 95-100 °C for 2h. The mixture was partitioned between ethyl acetate and aqueous acetic acid, the organic phase was concentrated, and the residue was subjected to chromatography on silica gel, eluting with 6:1 hexane/ethyl acetate, affording 337 mg of
10 3-mercapto-2-phenylpyridine. "-H NMR (CDC13) d 3.47 (s, 1) , 7.11 (m, 1) , 7.45 (m, 3) , 7.64 (m, 3) , 8.47, (d, 1, J=5) .
To a stirring solution of 3-mercapto-2-phenylpyridine (65 mg, 0.35 mmol) in DMF (1 mL) at 0 °C was added a solution of sodium
15 bis (trimethylsilyl) amide in THF (0.175 mL of 2 M solution, 0.35 mmol) . The resulting solution was transferred via cannula into a stirring solution of
(71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy) -3-cephem-4-carboxylate, benzhydryl ester (233 mg, 0.368
20 mmol) in DMF (5 mL) at -40 °C. After 30 min at -40 °C, the reaction was quenched by addition of 10% aqueous ammonium chloride solution, and the mixture was partitioned between water and ethyl acetate. The organic phase was dried over sodium sulfate, and was concentrated with a rotary - 25 evaporator. The crude product was purified by chromatography on silica gel (5% methanol/dichloromethane) to afford 165 mg of the title compound. NMR (CDC13) d 3.02 (d, 1, J=18) , 3.23 (d, 1, J=18) , 3.61 (d, 1, J=16) , 3.64 (d, 1, J=16) . 4.81 (d, 1, J=4) , 5.79 (dd, 1, J=4,9) , 6.20 (d, 1, J=9) , 7.01 (s, 1) , 7.2-7.6 (m, 21) , 7.67 (d, 1, J=10) , 8.61 (d, 1, J=5) .
EXAMPLE 168 (71?) -7- [ (phenylacetyl) amino] -3- (2-phenylpyrid-3-ylthio) -3-ce phem-4-carboxylic acid, trifluoroacetic acid salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-phenylpyrid-3-ylthio) -3-ce phem-4-carboxylate, benzhydryl ester (32 mg) and anisole (0.1 mL) in dichloromethane (1 mL) at 0 °C was added trifluoroacetic acid (1 mL) . After 0.5 h at 0 °C, the mixture was concentrated with a rotary evaporator. The crude product was triturated with ether, filtered, and dried to afford 20.1 mg of the title compound as an off-white solid. Η NMR (CD30D) d 3.22 (d, 1, J=18) , 3.49 (d, 1, J=18) , 3.55 (d, 1, J=16) , 3.60 (d, 1, J=16) , 5.03 (d, 1,
J=5) , 5.71 (d, 1, J=5) , 7.27 (m, 5) , 7.55 (m, 3) , 7.60 (m, 3) , 8.08 (d, 1, J=10) , 8.59 (d, 1, J=5) .
EXAMPLE 169 (71?) -7- [ (phenylacetyl)amino] -3- (2-cyanopyrid-3-ylthio) -3-cep hem-4-carboxylate, 4-methoxybenzyl ester
To a solution of 3-hydroxypicolinamide (2.00 g, 14.5 mmol) in phosphorus oxychloride (10 mL) at room temperature was added phosphorus pentachloride (6.00 g, 28.8 mmol) . The mixture was heated at reflux for 90 min, and the solvent was removed by distillation. The residue was poured into water, the aqueous solution was neutralized by addition of sodium bicarbonate, and was extracted with ethyl acetate. The organic phase was filtered through silica gel, the . Ill filtrate was concentrated, and the residue was subjected to flash chromatography on silica gel, affording 300 mg of 3-chloro-2-cyanopyridine. Η NMR (CDC13) d 7.50 (dd, 1, J=4.8, 8.4) , 7.89 (dd, 1, J=1.6, 8.4), 8.63 (dd, 1, J=1.6, 4.8) . 5 To a stirring solution of 4-methoxy-a-toluenethiol
(268 mg, 1.74 mmol) in DMF (1 mL) at 0°C was added a solution of sodium bis (trimethylsilyl) amide in THF (1.3 mL, 1 M, 1.3 mmol) . The resulting solution was added via cannula to a solution of 2-cyano-3-chloropyridine (200 mg,
10 1.45 mmol) in DMF (1 mL) at 0 °C. After stirring for 2 h at 0 °C, the mixture was poured into water and extracted with ethyl acetate, and the organic extracts were concentrated. The crude product was purified by column chromatography on silica gel, affording
15 2-cyano-3- (4-methoxybenzylthio)pyridine in quantitative yield. Η NMR (CDC13) d 3.80 (s, 3) , 4.20 (s, 2) , 6.83 (d, 2, J=10) , 7.22 (d, 2, J=10) , 7.34 (dd, 1, J=5, 8) , 7.66 (d, 1, J=8) , 8.50 (d, 1, J=5) . To a solution of this compound (245 mg, 0.96 mmol) and anisole (0.52 mL, 4.8 mmol) in
20 trifluoroacetic acid (1.5 mL) at 0 °C was added trifluoromethanesulfonic acid (0.25 mL, 2.8 mmol) . The mixture was allowed to warm to room temperature, and was stirred for 1 h. The solvent was removed with a rotary evaporator, and the residue was purified by flash - 25 chromatography on silica gel, affording 117 mg (90%) of 2-cyano-3-mercaptopyridine. Η NMR (CDC13) d 4.15 (s, 1) , 7.39 (dd, 1, J=5, 8) , 7.78 (d, 1, J=8) , 8.48 (d, 1, J=5) .
To a stirring solution of 2-cyano-3-mercaptopyridine (42 mg, 0.30 mmol) in dichloromethane (3 mL) at 0 °C was added a solution of sodium bis (trimethylsilyl) amide in THF (0.29 mL of 1 M solution, 0.29 mmol) . This solution was added dropwise via cannula to a stirring slurry of
(71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (200 mg, 0.34 mmol) in dichloromethane (1 mL) at -40 °C. The mixture was allowed to warm to room temperature and was stirred for 1 h, whereupon it was filtered through a pad of silica gel, washing with 1:1 dichloromethane/ethyl acetate. After concentrating the filtrate, the product was purified by flash chromatography on silica gel, affording 158 mg (90%) of the title compound. NMR (CDC13) d 3.15 (d, 1, J=18) , 3.60 (d, 1, J=18) , 3.63 (d, 1, J=16) , 3.68 (d, 1, J=16) , 3.80 (s, 3) , 5.04 (d, 1, J=4) , 5.15 (d, 1, J=12) , 5.27 (d, 1, J=12), 5.90 (dd, 1, J=4, 9) , 6.00 (d, 1, J=9) , 6.81 (d, 2, J=10) , 7.2-7.5 (m, 8), 7.70 (d, 1, J=8) , 8.55 (d, 1, J=5) .
EXAMPLE 170 (71?) -7- [ (phenylacetyl)amino] -3- (2-cyanopyrid-3-ylthio) -3-cep hem-4-carboxylic acid
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-cyanopyrid-3-ylthio) -3-cep hem-4-carboxylate, 4-methoxybenzyl ester (158 mg, 0.277 mmol) and anisole (0.4 mL) in dichloromethane (1 mL) at 0 °C was added trifluoroacetic acid (1 mL) . After 1 h at 0 °C, the solvent was removed with a rotary evaporator, and the residue was triturated with ether, filtered, and dried, affording 104 mg (83%) of the title compound. Η NMR (CDC13) d 3.15 (d, 1, J=18) , 3.54 (d, 1, J=18) , 3.65 (d, 1, J=16) , 3.69 (d, 1, J=16) , 5.07 (d, 1, J=4) , 5.89 (dd, 1, J= 4, 9) , 6.18 (d, 1, J=9) , 7.2-7.4 (m, 5) , 7.53 (dd, 1, J=5, 8) , 7.90 (d, 1, J=8) , 8.66 (d, 1, J=5) .
EXAMPLE 171
(71?) -7- [ (phenylacetyl)amino] -3- (1-phenylpyrazol-5-ylthio) r3- cephem-4-carboxylate, 4-methoxybenzyl ester
To a stirring solution of 1-phenylpyrazole (1.44 g, 10.0 mmol) in THF (7 mL) at -30 °C was added a solution of n-butyllithium in hexane (1.6 M, 6.88 mL, 11.0 mmol) . After 30 min at -30 °C, sulfur (352 mg, 11.0 mmol) was added, and the mixture was allowed to warm to 0 °C and stir for 7 h. Saturated aqueous ammonium chloride solution and 10% aqueous HCl were added, and the mixture was extracted with ethyl acetate. The organic layer was extracted with 5% aqueous potassium carbonate solution; the aqueous phase was acidified with 10% aqueous HCl and extracted with ethyl acetate, and the organic phase was dried (sodium sulfate) , filtered, and concentrated, affording 1.03 g of 5-mercapto-1-phenylpyrazole. Η NMR (CDC13) d 3.42 (s, 1) , 6.50 (d, 2) , 7.4-7.6 (m, 5) , 7.67 (d, 2) .
To a stirring solution of 5-mercapto-1-phenylpyrazole (396 mg, 2.25 mmol) in THF (3 mL) at 0 °C was added a solution of sodium bis (trimethylsilyl) amide in THF (2 M, 1.02 mL, 2.04 mmol) . After 5 min, the solvent was removed in vacuo, the residue was dissolved in DMF (3 mL) , and the resulting solution was added via syringe to a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (1.20 g, 2.04 mmol) in DMF (10 mL) at -50 °C. After 10 min at -50 °C, saturated aqueous ammonium chloride solution was added, and the mixture was poured into water at 0 °C and stirred for 1 h. The precipitated product was collected by filtration and was recrystallized from isopropanol, affording 856 mg (68%) of the title compound. lH NMR (CDC13) d 2.93 (d, 1, J=18) , 2.99 (d, 1, J=18) , 3.59 (d, 1, J=16) , 3.63 (d, 1, J=16) , 3.81 (s, 3) , 4.63 (d, 1, J=4) , 5.18 (d, 1, J=12) , 5.22 (d, 1, J=12) , 5.66 (dd, 1, J=4, 9) , 5.95 (d, 1, J=9) , 6.70 (d, 1, J=2) , 6.90 (d, 2, J=10) , 7.2-7.5 (m, 12) , 7.76 (d, 1, J=2) .
EXAMPLE 172 (7JF?) -7- [ (phenylacetyl)amino] -3- (l-phenylpyrazol-5-ylthio) -3- cephem-4-carboxylic acid
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (1-phenylpyrazol-5-ylthio) -3- cephem-4-carboxylate, 4-methoxybenzyl ester (80 mg, 0.12 mmol) and anisole (0.02 mL) in dichloromethane (2 mL) at 0 °C was added trifluoroacetic acid (0.5 mL) . After 2 h at 0 °C, the solvent was removed with a rotary evaporator, and the residue was subjected to chromatography on silica gel (3% acetic acid / ethyl acetate) , affording 49 mg (83%) of the title compound. Η NMR (acetone-d d 3.21 (s, 2) , 3.60 (d, 1, 3=16 ) , 3.65 (d, 1, J=16) , 4.96 (d, 1, J=4) , 5.71 (dd, 1, J= 4, 9) , 6.83 (d, 1, J=2) , 7.2-7.6 (m, 10) , 7.80 (d, 1, J=2) , 8.05 (d, 1, J=9) .
EXAMPLE 173 (71?) -7-amino-3- (l-phenylpyrazol-5-ylthio) -3-cephem-4-carboxy late, 4-methoxybenzyl ester, hydrochloride salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (l-phenylpyrazol-5-ylthio) -3- cephem-4-carboxylate, 4-methoxybenzyl ester (1.60 g, 2.61 5 mmol) and pyridine (0.53 mL, 6.5 mmol) in dry dichloromethane (10 mL) at 0 °C was added a solution of phosphorus pentachloride (1.09 g, 5.2 mmol) in dichloromethane (12 mL) . After 2.5 h at 0 °C, isobutanol (2.4 mL, 26 mmol) was added, and the mixture was stirred for
10 1.5 h. Water was added, the layers were separated, and the organic phase was washed with 5% aqueous sodium bicarbonate and 5% aqueous HCl, dried over sodium sulfate, filtered, and concentrated with a rotary evaporator. The resulting yellow oil was triturated with ether, filtered and dried to afford
15. 1.25 g (90%) of the title compound. Η NMR (CDCl3/CD3OD) d
2.74 (d, 1, J=18) , 3.17 (d, 1, J=18) , 4.65 (d, 1, J=4) , 4.84 (d, 1, J=4) , 6.65 (d, 1, J=2) , 6.75 (d, 2, J=9) , 7.20 (d, 2, J=9) , 7.2-7.4 (m, 5) , 7.64 (d, 1, J=2) .
20 EXAMPLE 174
(71?) -7- [ (cyanoacetyl) amino] -3- (1-phenylpyrazol-5-ylthio) -3-c ephem-4-carboxylate, 4-methoxybenzyl ester
To a stirring solution of (71?) -7-amino-3- (l-phenylpyrazol-5-ylthio) -3-cephem-4-carboxy - 25 late, 4-methoxybenzyl ester, hydrochloride salt (110 mg,
0.222 mmol) and cyanoacetic acid (36 mg, 0.42 mmol) in dry THF (4 mL) at -30 °C was added diisopropylethylamine (0.19 mL, 1.11 mmol) followed by phosphorus oxychloride (39.5 μL, 0.42 mmol) . After 1 h at -20 °C, the mixture was partitioned between water and ethyl acetate. The organic phase was washed with 5% aqueous sodium bicarbonate and 5% aqueous HCl, dried over sodium sulfate, and concentrated. The crude product was purified by chromatography on silica gel (1% methanol / dichloromethane) , affording 105 mg (85%) of the title compound. NMR (CDC13) d 2.93 (d, 1, J=18) , 3.09 (d, 1, J---18) , 3.40 (s, 2) , 3.81 (s, 3) , 4.67 (d, 1, J=4) , 5.20 (d, 1, J=12) , 5.22 (d, 1, J=12) , 5.61 (dd, 1, J=4, 9) , 6.72 (d, 1, J=2) , 6.87 (d, 2, J=10) , 7.10 (d, 1, J=9) , 7.3-7.5 (m, 7) , 7.76 (d, 1, J=2) .
EXAMPLE 175
(71?) -7- [ (cyanoacetyl)amino] -3- (l-phenylpyrazol-5-ylthio) -3-c ephem-4-carboxylic acid, sodium salt
To a stirring solution of (71?) -7- [ (cyanoacetyl) amino] -3- (l-phenylpyrazol-5-ylthio) -3-c ephem-4-carboxylate, 4-methoxybenzyl ester (105 mg, 0.187 mmol) and anisole (0.02 mL) in dichloromethane (2 mL) at 0 °C was added trifluoroacetic acid (0.5 mL) . After lh at 0 °C, the solvent was removed with a rotary evaporator. The residue was taken up in aqueous sodium bicarbonate, and was subjected to reverse-phase chromatography on C(18) , eluting with water followed by 20% acetonitrile / water. The fractions containing the desired product were lyophilized, affording 47.5 mg (50%) of the title compound. A sample was treated with HCl and dried for Η NMR analysis. Η NMR
(acetone-d d 3.24 (d, 1, J=18) , 3.30 (d, 1, J=18) , 3.79 (s, 2) , 5.06 (d, 1, J=4) , 5.76 (d, 1, J= ) , 6.87 (d, 1, J=2) , 7.4-7.6 (m, 5) , 7.82 (d, 1, J=2) . EXAMPLE 176
(71?) -7- [ (phenylacetyl) amino] -3- (1- (pyrid-2-yl)pyrazol-5-ylth io) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
To a stirring solution of 2-pyridylhydrazine (3.30 g, 30.2 mmol) and malonaldehyde bis (dimethylacetal) (4.96 g, 5 30.2 mmol) in absolute ethanol (60 mL) at room temperature was added concentrated HCl until the mixture was acidic (3 mL) . The mixture was heated at reflux for 3h, and was allowed to cool to room temperature. The mixture was concentrated with a rotary evaporator, and the residue was
10 triturated with ether and filtered. The remaining solid was dissolved in water, basified with saturated aqueous sodium bicarbonate, and extracted with ethyl acetate. The organic extracts were washed with water and brine, dried (sodium sulfate) , and concentrated with a rotary evaporator to
15. afford 3.10 g (70%) of 1- (pyrid-2-yl)pyrazole. Η NMR
(CDC1-) d 6.48 (dd, 1, J=l, 2) , 7.20 (dd, 1, J=8, 10) , 7.75 (d, 1, J=l) , 7.82 (dt, 1, J=2, 10) , 8.00 (d, 1, J=10) , 8.42 (dd, 1, J=2, 8) , 8.59 (d, 1, J=2) .
To a stirring solution of 1- (pyrid-2-yl)pyrazole
20 (1.00 g, 6.88 mmol) in THF (20 mL) at -78 °C was added a solution of n-butyllithium in hexanes (4.70 L, 1.6 M, 7.52 mmol) . After 15 min at -78 °C, sulfur (243 mg, 7.59 mmol) was added. The mixture was stirred at -78 °C for 15 min, and was warmed to 0 °C, and stirred for 30 min. Water was
25 added, and the THF was removed with a rotary evaporator.
The residue was washed with ethyl acetate, acidified to pH 6, and extracted with ethyl acetate. The latter organic extract was washed with brine, dried (sodium sulfate) , and concentrated with a rotary evaporator, affording 873 mg (73%) of 5-mercapto-l- (pyrid-2-yl)pyrazole. Η NMR (CDC13) d 6.20 (br s, 1) , 6.30 (d, 1, J=2) , 7.22 (dd, 1, J=8, 10) , 7.63 (d, 1, J=2) , 7.87 (dt, 1, J=2, 10) , 8.01 (d, 1, J=10) , 8.44 (dd, 1, J=2, 8) .
To a stirring solution of 5 5-mercapto-l- (pyrid-2-yl)pyrazole (95 mg, 0.540 mmol) in DMF (2 mL) at 0 °C was added a solution of sodium bis (trimethylsilylamide) in THF (0.40 mL, 1 M, 0.40 mmol) . This solution was added to a solution of (71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy)
10 -3-cephem-4-carboxylate, 4-methoxybenzyl ester (235 mg,
0.400 mmol) in DMF (12 mL) at -40 °C. After 20 min at -40 °C, the reaction was quenched by addition of saturated aqueous ammonium chloride solution (2 mL) and water. The mixture was extracted twice with ethyl acetate, and the
15. organic extracts were washed with water and brine, and were dried over sodium sulfate and concentrated with a rotary evaporator, affording 240 mg (98%) of the title compound, containing -10% of the D2 isomer. Η NMR (acetone-d6) d 3.54 (d, 1, J=18) , 3.63 (d, 1, J=16) , 3.70 (d, 1, J=16) , 3.78 (s,
20 3) , 3.80 (d, 1, J=18) , 5.17 (d, 1, J=10) , 5.21 (d, 1,
J=10) , 5.25 (d, 1, J=4) , 5.90 (dd, 1, J=4, 9) , 6.33 (d, 1, J=2) , 6.80 (d, 2, J=12) , 7.2-7.4 (m, 8) , 7.74 (d, 1, J=2) , 7.94 (d, 1, J=10) , 8.03 (dt, 1, J=2, 10) , 8.20 (d, 1, J=9) , 8.46 (dd, 1, J=2, 8) .
25
EXAMPLE 177
(71?) -7- [ (phenylacetyl)amino] -3- (1- (pyrid-2-yl)pyrazol-5-ylth io) -3-cephem-4-carboxylic acid
To a stirring suspension of (71?) -7- [ (phenylacetyl) amino] -3- (1- (pyrid-2-yl)pyrazol-5-ylth io) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (100 mg, 0.16 mmol) and anisole (0.6 mL) in dichloromethane (1.2 mL) at 0 °C was added trifluoroacetic acid (1.2 mL) . After 30 min at 0 °C, the solvent was removed with a rotary evaporator. The residue was dissolved in ethyl acetate, and was extracted with saturated aqueous sodium bicarbonate. The aqueous extracts were washed with ether, and were then acidified to pH 4-5 using aqueous HCl. This was extracted with ethyl acetate, and the organic extracts were washed with water and brine, and were dried (sodium sulfate) and concentrated. The resulting oil was triturated with hexane to afford the title compound (60 mg, 75%) as a pale yellow solid. Η NMR (acetone-d6) d 3.55 (d, 1, J=18) , 3.63 (d, 1, J=16) , 3.71 (d, 1, J=16) , 3.79 (d, 1, J=18) , 5.25 (d, 1, J=4) , 5.89 (dd, 1, J=4, 9) , 6.44 (d, 1, J=2) , 7.2-7.4 (m, 6) , 7.78 (d, 1, J=2) , 7.92 (d, 1, J=10) , 8.02 (dt, 1, J=2, 10) , 8.18 (d, 1, J=9) , 8.48 (dd, 1, J=2, 8) .
EXAMPLE 178 (71?) -7- [ (phenylacetyl)amino] -3- (2- (pyrazol-l-yl)pyrid-3-ylth io) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
A mixture of 2-chloro-3-nitropyridine (942 mg-, 5.94 mmol) , pyrazole (967 mg, 14.2 mmol) and powdered potassium carbonate (2 g) in DMF (10 mL) was stirred vigorously at room temperature for 4 days. The mixture was diluted with water, causing precipitation of a solid, which was filtered and dried, affording 670 mg (63%) of 3-nitro-2- (pyrazol-1-yl)pyridine. Η NMR (CDC13) d 6.51 (m, 1) , 7.37 (dd, 1 J=5, 8) , 7.76 (m, 1) 8.03 (dd, 1, J=2, 8) ,
SSSD/14615. V01 8 . 40 (m , 1 ) , 8 . 60 (dd , 1 , J=2 , 5 ) .
To a suspension of 3-nitro-2- (pyrazol-1-yl)pyridine (670 mg, 3.72 mmol) in ethanol (10 mL) was added a 1 M solution of SnCl2-H20 in 6 N aqueous HCl (14.5 mL, 14.5 mmol) . The mixture was heated at reflux for 10 min, and the solvent was removed with a rotary evaporator. 10% aqueous sodium hydroxide solution was added until most of the material dissolved, and the mixture was extracted with dichloromethane. The organic extracts were dried over sodium sulfate and concentrated to afford 376 mg (68%) of 3-amino-2- (pyrazol-1-yl)pyridine as a brown oil. Η NMR (CDC1,) d 5.62 (br s, 2) , 6.46 (m, 1) , 7.05 (dd, 1, J=5, 8) , 7.24 (dd, 1, J=2, 8) , 7.74 (m, 1) , 7.85 (dd, 1, J=2, 5) , 8.54 (d, 1, J=3) .
To a stirring solution of - 3-amino-2- (pyrazol-1-yl)pyridine (376 mg, 2.50 mmol) in 3 M aqueous HCl (4.3 mL) at 0 °C was added over 10 min a solution of sodium nitrite (181 mg, 2.63 mmol) in water (1 mL) . After 30 min, this solution was added to a solution of potassium ethyl xanthate (800 mg, 5.0 mmol) in water/saturated aqueous sodium bicarbonate (1:1, 22 mL) at 60 °C. The mixture was heated at 60 °C for 10 min, and was extracted with ethyl acetate, dried over sodium sulfate, and concentrated. The crude product was purified by silica gel chromatography (hexane/ethyl acetate 4:1) , affording 250 mg of the ethyl xanthate derivative. This material was dissolved in ethanol (4 mL) and was treated with 30% aqueous sodium hydroxide (2 mL) After 3 h, the mixture was neutralized by addition of 0.5 M citric acid solution, extracted with ethyl acetate, dried over sodium sulfate, and concentrated. The crude product was purified by silica gel chromatography, affording 166 mg (40%) of
3-mercapto-2- (pyrazol-1-yl)pyridine. *H NMR (CDC13) d 5.92 (br s, 1) , 6.47 (m, 1) , 7.08 (dd, 1, J=5, 8) , 7.72 (dd, 1, J=2, 8) , 7.78 (m, 1) , 8.21 (dd, 1, J=2, 5) , 8.42 (d, 1, J=3) .
To a solution of 3-mercapto-2- (pyrazol-1-yl)pyridine (77 mg, 0.435 mmol) in DMF (1 mL) at room temperature was added a 1 M solution of sodium bis (trimethylsilyl) amide in THF (0.42 mL, 0.42 mmol) . After 10 min, this solution was transferred via cannula into a solution of
(71?) -7- [ (phenylacetyl) amino] -3- (trifluoromethanesulfonyloxy) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (267 mg, 0.457 mmol) in DMF (2 mL) at -40 °C. After 1 h at -40 °C, - the mixture was poured into water and extracted with ethyl acetate. The organic extracts were washed with water, dried over sodium sulfate, and concentrated with a rotary evaporator. The residue was triturated with 1:1 hexane/ethyl acetate and dried, affording 291 mg of the title compound. Η NMR (CDC1,) d 3.10 (d, 1, J=18) , 3.52 (d, 1, J=18), 3.65 (d, 1, J=16) , 3.69 (d, 1, J=16), 3.75 (s, 3) , 5.01 (d, 1, J=4) , 5.26 (d, 1, J=12) , 5.28 (d, 1, J=12) , 5.89 (dd, 1, J=4, 9) , 6.05 (d, 1, J=9) , 6.50 (m, 1) , 6.72 (d, 2, J=10) , 7.13 (dd, 1, J=5, 8) , 7.20 (d, 2, J=10) , 7.2-7.4 (m, 5) , 7.59 (dd, 1, J=2, 8) , 7.78 (m, 1) , 8.28 (dd, 1, J=2, 5) , 8.40 (m, 1) .
EXAMPLE 179 (71?) -7- [(phenylacetyl)amino] -3- (2- (pyrazol-l-yl)pyrid-3-ylth io) -3-cephem-4-carboxylic acid, sodium salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2- (pyrazol-1-yl)pyrid-3-ylth io) -3-cephem-4-carboxylate, 4-methoxybenzyl ester (80 mg) and anisole (0.1 mL) in dichloromethane (1 mL) at 0 °C was added trifluoroacetic acid (1 mL) . After 30 min at 0 °C, the mixture was concentrated to an oil, and was partitioned between ether and saturated aqueous sodium bicarbonate. The aqueous phase was subjected to reverse phase chromatography on C(18) , eluting with water followed by acetonitrile/water. The fractions containing product were lyophilized, affording 43 mg of the title compound. lH NMR (D20) d 3.10 (d, 1, J=18) , 3.50 (d, 1, J=18), 3.64 (d, 1, J=16) , 3.69 (d, 1, J=16) , 5.10 (d, 1, J=4) , 5.62 (d, 1, J=4) , 6.61 (m, 1) , 7.2-7.4 (m, 5) , 7.52 (m, 1) , 7.82 (m, 1) , 7.93 (br d, J=8) , 8.14 (m, 1) , 8.37 (m, 1) .
EXAMPLE 180 (71?) -7- [ (phenylacetyl) amino] -3- [2- (N,N' - (bis) tert-butyloxyca rbonylguanidinomethyl)phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
To a stirring solution of 2-mercaptobenzyl alcohol (1.00 g, 7.10 mmol) in dichloromethane (20 mL) at 0 °C was added methyl acrylate (0.70 mL, 7.70 mmol) followed by triethylamine (50 μL, 0.35 mmol) . The mixture was allowed to warm to room temperature and stir for 4 h, at which point the solvent was removed with a rotary evaporator. The residue was subjected to column chromatography on silica gel (0-10% ethyl acetate in dichloromethane) , affording 1.40 g (86%) of methyl 3- (2-hydroxymethylphenylthio)propionate. H NMR (CDCl,) d 2.63 (t, 2, J=7) , 3.18 (t, 2, J=7) , 3.68 (s, 3) , 4.79 (s, 2) , 7.27 (m, 2) , 7.43 (m, 2) . To a stirring solution of methyl 3- (2-hydroxymethylphenylthio)propionate (1.40 g, 5.10 mmol) , triphenylphosphine (2.00 g, 7.60 mmol) , and N,N' - (bis) tert-butyloxycarbonylguanidine (2.60 g, 10.2 mmol) in THF (30 mL) at 0 °C was slowly added diisopropyl azodicarboxylate (1.50 mL, 7.60 mmol) . The mixture was allowed to warm to room temperature and was stirred for 5 h. The solvent was removed with a rotary evaporator, and the residue was subjected to column chromatography on silica gel, affording 1.62 g (68%) of methyl
3- [2- (N,N' - (bis) ert-butyloxycarbonylguanidinomethyl)phenylt hio]propionate. Η NMR (CDC1 dl.25 (s, 9) , 1.46 (s, 9) , 2.62 (t, 2, J=7) , 3.13 (t, 2, J=7) , 3.69 (s, 3) , 5.39 (s, 2) , 7.0-7.4 (m, 4) , 9.42 (br s, 1) , 9.53 (br s, 1) .
To a stirring solution of methyl 3- [2- (N,N' - (bis) tert-butyloxycarbonylguanidinomethyl)phenylt hio] propionate (63 mg, 0.13 mmol) in DMF (1 mL) at room temperature was added potassium tert-butoxide (15 mg, 0.13 mmol) . After 4 h, this mixture was transferred by cannula into a stirring solution of
(71?) -7- [ (phenylacetyl) amino] -3-trifluoromethanesulfonyloxy-3 -cephem-4-carboxylate, 4-methoxybenzyl ester (79 mg, 0.13 mmol) in DMF (0.5 mL) at -40 °C. The mixture was stirred for 40 min at -40 °C, and was quenched by addition of saturated ammonium chloride solution. The mixture was extracted with ethyl acetate/hexane, and the organic phase was dried over sodium sulfate and concentrated with a rotary evaporator. Column chromatography on silica gel (50% ethyl acetate in hexane) afforded 44 mg (40%) of the title compound. -H NMR (CDCl,) dl.28 (s, 9) , 1.42 (s, 9) , 3.05 (d, 1, J=18) , 3.33 (d, 1, J=18) , 3.59 (d, 1, J=16) , 3.62 (d, 1, J=16) , 3.81 (s, 3) , 5.00 (d, 1, J=4), 5.07 (d, 1, J=18), 5.26 (s, 2) , 5.32 (d, 1, J=18) , 5.78 (dd, 1, J=4,9) , 6.15 (d, 1, J=9) , 6.89 (d, 2, J=12) , 7.03 (d, 1, J=10) , 7.2-7.4 (m, 9) , 7.52 (d, 1, J=10) , 9.39 (br s, 1) , 9.44 (br s, 1) .
EXAMPLE 181
(71?) -7- [ (phenylacetyl)amino] -3- [2- (guanidinomethyl)phenylthi o] -3-cephem-4-carboxylate, inner salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- [2- (N,N' - (bis) tert-butyloxyca rbonylguanidinomethyl)phenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester (44 mg, 0.055 mmol) and anisole (0.2 mL) in dichloromethane (0.4 mL) at 0 °C was added trifluoroacetic acid (1 mL) . The mixture was allowed to warm to room temperature and stir for 40 min. The solvent was removed in vacuo, and the residue was triturated with ether and filtered, affording the title compound (25 mg, 95%) as a light yellow powder. >H NMR (CD3OD) d3.10 (d, 1, J=17) , 3.41 (d, 1, J=17) , 3.54 (d, 1, J=12) , 3.58 (d, 1, J=12) , 4.48 (s, 2) , 5.04 (d, 1, J=5) , 5.59 (d, 1, J=5) , 7.2-7.4 (m, 9) .
EXAMPLE 182
(71?) -7- [ (phenylacetyl)amino] -3- (2-chloromethylphenylthio) -3- cephem-4-carboxylate, 4-methoxybenzyl ester To dry DMF (100 mL) at 0 °C was added thionyl chloride (1.3 mL, 17.8 mmol) . After 30 min at 0 °C, to this solution was added
(71?) -7- [ (phenylacetyl) amino] -3- (2-hydroxymethylphenylthio) -3 -cephem-4-carboxylate, 4-methoxybenzyl ester (6.90 g, 12.0 mmol) . The resulting solution was stirred at 0 °C for 30 min, and was poured into ice water (800 mL) . After 30 min, the mixture was filtered, and the precipitate was washed with water and air-dried. The resulting pale brown solid was taken up in dichloromethane (500 mL) , washed with water and saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated with a rotary evaporator. The residue was triturated with ether, filtered and dried to afford the title compound (4.35 g, 61%) as an off-white solid. Η NMR (CDCl3) d 3.08 (d, 1, J=18) , 3.25 (d, 1, J=18) , 3.60 (d, 1, J=16) , 3.63 (d, 1, J=16) , 3.81 (s, 3) , 4.75 (d, 1, J=14) , 4.79 (d, 1, J=14) , 4.94 (d, 1, J=4) , 5.25 (s, 2) , 5.75 (dd, 1, J=4,9) , 6.04 (d, 1, J=9) , 6.86 (d, 2, J=10) , 7.2-7.6 (m, 11) .
EXAMPLE 183
(71?) -7-amino-3- (2-chloromethylphenylthio) -3-cephem-4-carboxy late, 4-methoxybenzyl ester, trifluoroacetate salt
To a stirring solution of (71?) -7- [ (phenylacetyl) amino] -3- (2-chloromethylphenylthio) -3- cephem-4-carboxylate, 4-methoxybenzyl ester (11.6 g, 19.5 mmol) and pyridine (3.20 mL, 39.6 mmol) in dichloromethane (50 mL) at 0 °C was added slowly a solution of phosphorus pentachloride (6.40 g, 30.7 mmol) in dichloromethane (75 mL) . After 2 h at 0 °C, isobutanol (1.82 mL, 19.8 mmol) was added. The mixture stirred for 2 h at 0 °C, dichloromethane was added (500 mL) , and the resulting solution was washed with saturated aqueous sodium bicarbonate (2 X 400 mL) , water (2 X 400 mL) , IM aqueous HCl (2 X 400 mL) , and brine (2 X 400 mL) . The organic phase was dried (sodium sulfate) , trifluoroacetic acid (1.6 mL, 20.8 mmol) was added, and the mixture was concentrated with a rotary evaporator. The resulding beige solid was triturated with ether and dried, affording the title compound (6.53 g, 57%) as a pale brown solid. lH NMR (CDCl3/CD3OD) d 2.85 (d, 1, J=18) , 3.34 (d, 1, J=18) , 3.72 (s, 3) , 4.62 (d, 1, J=8) , 4.74 (d, 1, J=8) , 4.74 (m, 1) , 5.06 (m, 1) , 5.17 (s, 2) , 6.80 (d, 2, J=10) , 7.2-7.5 (m, 6) .
EXAMPLE 184 (71?) -7- [ ( (2-N,N' - (bis) - ert-butyloxycarbonylguanidinoethylth io) acetyl)amino] -3- [2- (chloromethyl)phenylthio] -3-cephem-4-c arboxylate, 4-methoxybenzyl ester
To a stirring solution of N,N' - (bis) -di-tert-butyloxycarbonyl-S-methylisothiourea (2.37 g, 8.17 mmol) and 2-mercaptoethylamine hydrochloride (0.88 g, 7.78 mmol) at room temperature in THF (40 mL) was added triethylamine (1.2 mL, 8.56 mmol) . The reaction was heated at 45-50 °C for 7 h, after which the solvent was removed with a rotary evaporator. The residue was triturated with hexane/ethyl acetate, triethylamine hydrochloride was removed by filtration. The filtrate was concentrated and subjected to column chromatography on silica gel (2-5% ethyl acetate/hexanes) , affording 1.08 g (44%) of 2- (N,N' -di-tert-butyloxycarbonylguanidino) ethanethiol. Η NMR (CDC13) d 1.43 (t, 1, J=8) , 1.48 (s, 18) , 2.73 (dt, 2, J=8, 6) , 3.63 (q, 2, J=6) , 8.67 (br s, 2) .
To a stirring solution of 2- (N,N' -di- tert-butyloxycarbonylguanidino) ethanethiol (1.08 g, 3.37 mmol) and methyl bromoacetate (516 mg, 3.37 mmol) in THF (10 mL) at 0 °C was added triethylamine (0.56 mL, 4.04 mmol) . The mixture was allowed to warm to room temperature and was stirred overnight. The mixture was partitioned between ethyl acetate and water, and the aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with water and brine, dried over sodium sulfate, and concentrated. The residue was subjected to column chromatography on silica gel (5-10% ethyl acetate/hexane) , affording 1.24 g (95%) of methyl [2- (N,N' -di- tert-butyloxycarbonylguanidino) ethyl] thioacetate . Η NMR (CDC13) d 1.48 (s, 18) , 2.83 (t, 2, J=8) , 3.26 (s,. 2) , 3.65 (q, 2, J=8) , 3.73 (s, 3) , 8.59 (br s, 2) .
To a stirring solution of methyl [2- (N,N' -di- tert-butyloxycarbonylguanidino) ethyl] thioacetate (1.24 g, 3.17 mmol) in ethanol (15 mL) at room temperature (15 mL) was added 10% aqueous sodium hydroxide solution (3 mL) . After 2 h, The mixture was concentrated, and the residue was treated with 10% aqueous HCl solution. The resulting mixture was extracted with ethyl acetate, and the organic extracts were washed with water and brine, dried over sodium sulfate, and concentrated to afford 1.06 g of [2- (N.N' -di- tert-butyloxycarbonylguanidino) ethyl] thioacetic acid as a white solid. Η NMR (CDC13) d 1.50 (s, 18) , 2.89 (t, 2, J=8) , 3.34 (s, 2) , 3.72 (m, 2) , 8.82 (br s, 2) . To a stirring solution of
[2- (N,N' -di- ert-butyloxycarbonylguanidino) ethyl] thioacetic acid (153 mg, 0.406 mmol) and
(71?) -7-amino-3- (2-chloromethylphenylthio) -3-cephem-4-carboxy late, 4-methoxybenzyl ester, trifluoroacetate salt (200 mg, 0.334 mmol) at -20 °C in THF (8 mL) was added diisopropylethylamine (233 μL, 1.34 mmol) followed by phosphorus oxychloride (39 μL, 0.406 mmol) . After 2 h, the mixture was partitioned between ethyl acetate and water, the layers were separated, and the organic layer was washed with saturated aqueous sodium bicarbonate solution, water, 10% aqueous HCl solution, and brine. After drying over sodium sulfate, the mixture was concentrated with a rotary evaporatory, and the residue was subjected to chromatography on silica gel (0-3% MeOH/dichloromethane) , affording 228 mg
(82%) of the title compound. lH NMR (CDC13) d 1.51 (s, 18) , 2.80 (t, 2, J=8) , 3.22 (d, 1, J=18) , 3.30 (d, 1, J=18) , 3.34
(s, 2) , 3.62 (m, 2) , 3.82 (s, 3) , 4.80 (s, 2) , 5.01 (d, 1, J=4) , 5.31 (s, 2) , 5.75 (dd, 1, J-4, 9) , 6.90 (d, 2, J=10) , 7.3-7.6 ( , 7) , 8.60 (br s, 2) .
EXAMPLE 185
(71?) -7- [ ( (2-N,N' - (bis) - ert-butyloxycarbonylguanidinoethylth io)acetyl) amino] -3- [2- (iodomethyl)phenylthio] -3-cephem-4-car boxylate, 4-methoxybenzyl ester To a stirring solution of (71?) -7- [ ( (2-N,N' - (bis) - tert-butyloxycarbonylguanidinoethyl thio) acetyl) amino] -3- [2- (chloromethyl)phenylthio] -3-cephem-4 -carboxylate, 4-methoxybenzyl ester (228 mg, 0.273 mmol) at room temperature in acetonitrile (3 mL) was added sodium iodide (205 mg, 1.34 mmol) . After stirring overnight, the mixture was partitioned between water and ethyl acetate, and the organic layer was washed with water and brine. The solvent was removed with a rotary evaporator, affording 239 mg (94%) of the title compound. Η NMR (CDCl3) d 1.51 (s, 18) , 2.80 (t, 2, J=8) , 3.28 (d, 1, J=18) , 3.35 (s, 2) , 3.37 (d, 1, J=18) , 3.62 (m, 2) , 3.82 (s, 3) , 4.60 (d, 1, J=9) , 4.63 (d, 1, J=9) , 5.06 (d, 1, J=4) , 5.29 (s, 2) , 5.77 (dd, 1, J-4, 9) , 6.89 (d, 2, J=10) , 7.3-7.6 (m, 7) , 8.60 (br s, 2) . EXAMPLE 186
(71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io)acetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-ce phem-4-carboxylate, 4-methoxybenzyl ester
To a stirring solution of (71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io) acetyl) amino] -3- [2- (iodomethyl)phenylthio] -3-cephem-4-car boxylate, 4-methoxybenzyl ester (239 mg, 0.258 mmol) at room temperature in acetonitrile (2 mL) was added thiourea (19 mg, 0.258 mmol) . After 4 h, the solvent was removed with a rotary evaporator, and the residue was triturated with ether, filtered, and dried, affording 240 mg (93%) of the title compound. lH NMR (CDC13) d 1.48 (s, 18) , 2.81 (m, 2) , 3.21 (d, 1, J=18) , 3.40 (s, 2), 3.42 (d, 1, J=18) , 3.60 (m, 2) , 3.80 (s, 3) , 4.70 (d, 1, J=ll) , 4.87 (d, 1, J=ll) , 5.12 (d, 1, J=4) , 5.27 (s, 2) , 5.56 (dd, 1, J-4, 9) , 6.50 (br s, 1) , 6.89 (d, 2, J=10) , 7.3-7.6 (m, 6) , 8.15 (d, 1, J=9) , 8.50 (v br s, 1) , 8.66 (br s, 1) .
EXAMPLE 187
(71?) -7- [ ( (2-guanidinoethylthio)acetyl) amino] -3- [2- (isothiour oniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
To a mixture of (71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io) acetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-ce phem-4-carboxylate, 4-methoxybenzyl ester (240 mg, 0.240 mmol) and anisole (100 μL) at room temperature was added trifluoroacetic acid (1.5 mL) . After 1.5 h, the volatiles were removed in vacuo, and the residue was triturated with ether. The resulting solid was dried, and was suspended in water (50 mL) , filtered, and lyophilized to afford 131 mg (82%) of the title compound. W NMR (D20) d 2.83 (t, 2, J=7) , 3.25 (d, 1, -7=18) , 3.38 (s, 2) , 3.42 (t, 2, J=7) , 3.48 (d, 1, J=18) , 4.59 (s, 2) , 5.20 (d, 1, J=4) , 5.59 (d, 1, J=4) , 7.4-7.6 (m, 4) .
Under substantially the same procedures as in Examples 184-187, the items of the following Examples 188-219 can similarly be obtained.
EXAMPLE 188 (71?) -7- [( (3-guanidinopropylthio)acetyl)amino] -3- [2- (isothiou roniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt EXAMPLE 189
(71?) -7- [ ( (4-guanidinobutylthio)acetyl) amino] -3- [2- (isothiour oniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 190
(71?) -7- [ ( (2-N,N! -dimethylguanidinoethylthio) acetyl)amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 191
(71?) -7- [ ( (3-N,N' -di ethylguanidinopropylthio)acetyl)amino] -3 - [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate
, trifluoroacetic acid salt
EXAMPLE 192
(71?) -7- [( (4-N,N' -dimethylguanidinobutylthio)acetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 193
(71?) -7- [ ( (2-isothiouronium) ethylthioacetyl)amino] -3- [2- (isot hiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 194
(71?) -7- [ ( (3-isothiouroniumpropyl) thioacetyl)amino] -3- [2- (iso thiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt EXAMPLE 195
(71?) -7- [ ( (4-isothiouroniumbutyl) thioacetyl) amino] -3- [2- (isot hiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 196
(71?) -7- [ ( (2-N,N* -dimethylisothiouronium) ethylthioacetyl) amin ] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxy late, trifluoroacetic acid salt
EXAMPLE 197
(71?) -7- [ ( (3-N,N"dimethylisothiouroniumpropyl) thioacetyl) amin ] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxy late, trifluoroacetic acid salt
EXAMPLE 198
(71?) -7- [ ( (4-N,N" -dimethylisothiouroniumbutyl) thioacetyl)amin o] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxy late, trifluoroacetic acid salt
EXAMPLE 199
(71?) -7- [((2- tetramethylisothiouronium) ethylthioacetyl) amino] -3- [2- (isoth iouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt EXAMPLE 200
(71?) -7- [ ( (3-tetramethylisothiouroniumpropyl) thioacetyl) amino
] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxyl ate, trifluoroacetic acid salt
EXAMPLE 201
(71?) -7- [ ( (4-tetramethylisothiouroniumbutyl) thioacetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxyla te, trifluoroacetic acid salt
EXAMPLE 202
(71?) -7- [ (2- ( (iminomethyl) amino) ethylthioacetyl) amino] -3- [2- ( isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 203
(71?) -7- [ (2- ( (N-methyliminomethyl)amino) ethylthioacetyl) amino ] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxyl ate, trifluoroacetic acid salt
EXAMPLE 204
(71?) -7- [ (3- ( (iminomethyl)amino)propylthioacetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 205
(71?) -7- [ (3- ( (N-methyliminomethyl)amino)propylthioacetyl)amin o] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxy late, trifluoroacetic acid salt EXAMPLE 206
(71?) -7- [ (4- ( (iminomethyl) amino)butylthioacetyl) amino] -3- [2- ( isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 207
(71?) -7- [ (4- ( (N-methyliminomethyl) amino)butylthioacetyl) amino ] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxyl ate, trifluoroacetic acid salt
EXAMPLE 208
(71?) -7- [ ( (2-amidinium) ethylthioacetyl) amino] -3- [2- (isothiour oniummethyl)phenylthio] -3-cephem-4-carboxyl te, trifluoroacetic acid salt
EXAMPLE 209
(71?) -7- [ ( (3-amidiniumpropyl) thioacetyl) amino] -3- [2- (isothiou roniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 210
(71?) -7- [ ( (4-amidiniumbutyl) thioacetyl) amino] -3- [2- (isothiour oniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt EXAMPLE 211 (71?) -7- [ (2- (N,N' -dimethylamidiniu ) ethylthioacetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 212
(71?) -7- [ ( (3- (N,N'dimethylamidinium)propyl) thioacetyl) amino] - - [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylat e, trifluoroacetic acid salt
EXAMPLE 213
(71?) -7- [ ( (4- (N,N! -dimethylamidinium)butyl) thioacetyl) amino] - 3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylat e, trifluoroacetic acid salt
EXAMPLE 214
(71?) -7- [ ( (2- (tetramethylamidinium) ethyl) thioacetyl) amino] -3- [2- (isothiouroniummethyl) henylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 215
(71?) -7- [ ( (3- (tetramethylamidinium)propyl) thioacetyl) amino] -3 - [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate
, trifluoroacetic acid salt
EXAMPLE 216
(71?) -7- [( (4- (tetramethylamidinium)butyl) thioacetyl) amino] -3- [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt EXAMPLE 217
(71?) -7- [ ( (2-amidinium)methylthioacetyl)amino] -3- [2- (isothiou roniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 218
(71?) -7- [ ( (2-N,N* -dimethylamidinium)methylthioacetyl)amino] -3 - [2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate
, trifluoroacetic acid salt
EXAMPLE 219
(71?) -7- [ ( (2-tetramethylamidinium) ethylthioacetyl)amino] -3- [ 2- (isothiouroniummethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 220
(71?) -7- [ (phenylacetyl)amino] -3- (2,4- (bis)hydroxymethylphenyl thio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example 161, but sodium 2,4- (bis) hydroxymethylbenzenethiolate was used.
EXAMPLE 221
(71?) -7- [ (phenylacetyl)amino] -3- (2,4- (bis)chloromethylphenylt hio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester This was prepared by a procedure similar to that for Example 182, but using
(71?) -7- [ (phenylacetyl) amino] -3- (2,4- (bis) hydroxymethylphenyl thio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material. EXAMPLE 222
(71?) -7- [ (phenylacetyl)amino] -3- (2,4- (bis) iodomethylphenylthi o) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example
185, but using (71?) -7- [ (phenylacetyl) amino] -3- (2,4- (bis) chloromethylphenylt hio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 223 (71?) -7- [ (phenylacetyl)amino] -3- (2,4- (bis) isothiouroniummethy lphenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example
186, but using
(71?) -7- [ (phenylacetyl) amino] -3- (2,4- (bis) iodomethylphenylthi o) -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material .
EXAMPLE 224 (71?) -7- [ (phenylacetyl)amino] -3- (2,4- (bis) isothiouroniummethy lphenylthio) -3-cephem-4-carboxylate, trifluoroacetic acid salt
This was prepared by a procedure similar to that for Example
187, but using
(71?) -7- [ (phenylacetyl) amino] -3- (2,4- (bis) isothiouroniummethy lphenylthio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 225 (71?) -7-amino-3- (2,4- (bis)chloromethylphenylthio) -3-cephem-4- carboxylate, 4-methoxybenzyl ester, trifluoroacetatic acid salt
This was prepared by a procedure similar to that for Example
183, but using
(71?) -7- [ (phenylacetyl) amino] -3- (2,4- (bis) chloromethylphenylt hio) -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 226 (71?) -7- [ ( (2-N,N' - (bis) - ert-butyloxycarbonylguanidinoethylth io)acetyl)amino] -3- [2,4- (bis)chloromethylphenylthio] -3-cephe m-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example
184, but using
(71?) -7-amino-3- (2,4- (bis) chloromethylphenylthio) -3-cephem-4- carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 227 (71?) -7- [ ( (2-N,N' - (bis) -terfc-butyloxycarbonylguanidinoethylth io) acetyl)amino] -3- [2,4- (bis) iodomethylphenylthio] -3-cephem- 4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example
185, but using
(71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io) acetyl) amino] -3- [2,4- (bis) chloromethylphenylthio] -3-cephe m-4-carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 228 (71?) -7- [ ( (2-N,N' - (bis) - ert-butyloxycarbonylguanidinoethylth io)acetyl)amino] -3- [2,4- (bis) isothiouroniummethylphenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester
This was prepared by a procedure similar to that for Example
186, but using
(71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io) acetyl) amino] -3- [2,4- (bis) iodomethylphenylthio] -3-cephem- 4-carboxylate, 4-methoxybenzyl ester as starting material.
EXAMPLE 229 (71?) -7- [ ( (2-guanidinoethylthio)acetyl)amino] -3- [2,4- (bis) iso thiouroniummethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
This was prepared by a procedure similar to that for Example
187, but using
(71?) -7- [ ( (2-N,N' - (bis) -tert-butyloxycarbonylguanidinoethylth io) acetyl) amino] -3- [2,4- (bis) isothiouroniummethylphenylthio] -3-cephem-4-carboxylate, 4-methoxybenzyl ester as starting material .
Under substantially the same procedures as in Examples 226-229, the items of the following Examples 230-233 were similarly obtained.
EXAMPLE 230
(71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino) ace tamido] -3- [2,4- (bis) isothiouroniummethylphenylthio] -3-cephem
-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 231
(71?) -7- [ (cyanomethylthioacetyl) amino] -3- [2,4- (bis) isothiouro niummethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 232
(71?) -7- [ (methylthioacetyl) amino] -3- [2,4- (bis) isothiouroniumm ethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 233
(71?)-7-[(Z)-2- (Aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- [2,4- (bis) isothiouroniummethylphenylthio] -3-cephem-4-carb oxylate, trifluoroacetic acid salt
EXAMPLE 234
(71?) -7- [ ( (2- (isothiouronium) ethylthio) acetyl) amino] -3- [2,4- ( bis) isothiouroniummethylphenylthio] -3-cephem-4-carboxylate, trifluoroacetic acid salt
EXAMPLE 235
(7R) -7- [ (phenylacetyl) amino] -3- [2- (amidinomethylthiomethyl)p henylthio] -3-cephem-4-carboxylic acid
A solution of (7R) -7- [ (phenylacetyl) amino] -3- (2-chloromethylphenylthio) -3- cephem-4-carboxylate, 4-methoxybenzyl ester (0.50 g) and anisole (0.1 mL) in trifluoroacetic acid (1 mL) was stirred at 0 °C for 45 min. The mixture was concentrated to an oil, and was triturated with diethyl ether/hexane, filtered, and dried, affording 0.40 g of (7R) -7- [ (phenylacetyl) amino] -3- [2- (chloromethyl) henylthio] - 3-cephem-4-carboxylic acid. 1H NMR (CDC13) d 3.20 (d, 1, J=17) , 3.29 (d, 1, J=17) , 3.54 (d, 1, J=15) , 3.58 (d, 1, J=15) , 4.80 (d, 1, J=ll) , 4.86 (d, 1, J=ll) , 5.10 (d, 1, J=5) , 5.61 (d, 1, J=5) , 7.2-7.6 (m, 8) . A mixture of this carboxylic acid intermediate (50 mg,
0.1 mmol) , amidinomethylthiol hydrochloride (24 mg, 0.2 mmol) , sodium bicarbonate (34 mg, 0.4 mmol) and sodium iodide (30 mg, 0.2 mmol) in water (5 mL) was stirred at room temperature for 16 h. The resulting precipitate was filtered, washed with water, and vacuum-dried, affording 30 mg of the title compound. Η NMR (DMSO-d6) d3.07 (d, 1, J=17) , 3.42 (d, 1, J=17) , 3.47 (d, 1, J=14) 3.54 (d, 1, J=14) , 4.07 (d, 1, J=13) , 4.18 (d, 1, J=13) , 5.10 (d, 1, J=5) , 5.54 (dd, 1, J= 5, 8), 7.2-7.5 (m, 8) 9.09 (d, 1, J=8) , 9.40 (br s, 1) .
EXAMPLE 236 (7R) -7-{ [ (2-guanidinoethylthio)acetyl] amino}-3- [2- (amidinium methylthiomethyl)phenylthio] -3-cephem-4-carboxylic acid, acetate salt
A solution of (7R) -7- { [2- [N,N' -bis- (t-butyloxycarbonyl) guadinino] ethylthio ] acetyl] amino}-3- (2-chloromethylphenylthio) -3-cephem-4-carbo xylate, 4-methoxybenzyl ester (315 mg, 0.376 mmol) and sodium iodide (300 mg, 2.0 mmol) in acetone (10 mL) was stirred at 30 °C for 1 h. The mixture was concetrated to a volume of 1 mL and was partitioned between dichloromethane and 0.5% aqueous sodium thiosulfate. The organic layer was dried over sodium sulfate and concentrated to dryness, affording
(7R) -7-{ [2- [N.N' -bis- (t-butyloxycarbonyl) guadinino] ethylthio acetyl] amino} -3- (2-iodomethylphenylthio) -3-cephem-4-carboxyl ate, 4-methoxybenzyl ester. The iodide was dissolved in ethanol/acetonitrile/water
(40 mL/10 mL/2 mL) . Amidinomethylthiol hydrochloride (118 mg, 0.95 mmol) and 2,6-lutidine (0.10 mL, 0.95 mmol) were added and the mixture was stirred at room temperature for 7 h. Trifluoroacetic acid (75 mL, 0.95 mmol) was added, and the mixture was concentrated. The mixture was partitioned between dichloromethane and water. The organic phase was concentrated, and the residue was purified by silica gel chromatography (5% methanol/dichloromethane) to afford 193 mg of (7R) -7- { [2- [N,N' -bis- (t-butyloxycarbonyl) guadinino] ethylthio acetyl] amino}-3- [2- (amidiniummethylthiomethyl)phenylthio] -3- cephem-4-carboxylate, 4-methoxybenzyl ester, hydroiodide salt. Η NMR (CDC13) dl.30 (s, 9) , 1.50 (s, 9) , 2.81 (m, 2) , 3.21 (s, 2) , 3.31 (d, 1, J=15) , 3.38 (d, 1, J=15) , 3.49 (s, 2) , 3.74 (m, 2) , 3.80 (s, 3) , 3.88 (d, 1, J=13) , 4.11 (d, 1, J=13) , 5.01(d, 1, J=5) , 5.23 (d, 1, J=12) , 5.27 (d, 1, J=12) , 5.64 (dd, 1, J=5, 8) 6.88 (d, 2, J=9) , 7.3-7.5 (m, 6)', 8.03 (br s, 2) , 8.25 (d, 1, J=8) , 10.32 (br s, 2) . A mixture of (7R) -7-{ [2- [N,N' -bis- (t-butyloxycarbonyl)guadinino] ethylthio acetyl] amino} -3- [2- (amidiniummethylthiomethyl)phenylthio] -3- cephem-4-carboxylate, 4-methoxybenzyl ester, hydroiodide salt (193 mg) and anisole (0.3 mL) in trifluoroacetic acid (3 mL) was stirred at room temperature for 45 min. The mixture was concentrated under reduced pressure, redissolved in IM aqueous ammonium acetate solution (100 mL) , and filtered. The filtrate was subjected to HP-20 reverse-phase column chromatography, washing with water and then eluting with 10% acetonitrile/water. The fractions containing the desired product were freeze-dried, affording 36 mg of the title compound. Η NMR (D20) d 1.91 (s, 3) , 2.84 (t, 2, J=6) , 3.12 (d, 1, J=17) , 3.43 (t, 2, J=6) , 3.50 (d, 1, J=17) , 3.56 (s, 2) , 4.02 (s, 2) , 5.19 (d, 1, J=5) , 5.62 (d, 1, J=5) , 7.3-7.5 (m, 4) .
Under substantially the same procedures as in Examples 235-236, the items of the following Examples 237-249 were similarly obtained.
EXAMPLE 237
(71?) -7- [ (Phenylacetyl)amino] -3- (2-guanidinoethylthiomethylph enylthio) -3-cephem-4-carboxylate, inner salt
EXAMPLE 238
(71?) -7- [ ( (2-guanidinoethylthio)acetyl) amino] -3- (2-formamidoy lmethylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt EXAMPLE 239 (71?) -7- [( (2-guanidmoethylthio) acetyl)amino] -3- (2-guanidinoe thylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt
EXAMPLE 240 (71?) -7- [( (2-guanidmoethylthio) acetyl)amino] -3- (2-dimethylam inoethylthiomethylphenylthio) -3-cephem-4-carboxylate, inner salt
EXAMPLE 241 (71?)-7-[(Z)-2- (aminothiazol-4-yl) -2- (hydroxyimino)acetamido] -3- (2-amidinomethylthiomethylphenylthio) -3-cephem-4-carboxyl ate, inner salt
EXAMPLE 242 (71?)-7-[(Z)-2- (aminothiazol-4-yl) -2- (hydroxyimino)acetamido] -3- (2-formamidoylmethylthiomethylphenylthio) -3-cephem-4-carb oxylate, inner salt
EXAMPLE 243 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino)acetamido] -3- (2-guanidinoethylthiomethylphenylthio) -3-cephem-4-carboxy late, inner salt
EXAMPLE 244 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- (2-dimethylaminoethylthiomethylphenylthio) -3-cephem-4-car boxylate, inner salt
EXAMPLE 245 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino)ace tamido] -3- (2-amidinomethylthiomethylphenylthio) -3-cephem-4-c arboxylate, inner salt
EXAMPLE 246 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino)ace tamido] -3- (2-formamidoylmethylthiomethylphenylthio) -3-cephem
-4-carboxylate, inner salt
EXAMPLE 247 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino)ace tamido] -3- (2-guanidinoethylthiomethylphenylthio) -3-cephem-4- carboxylate, inner salt
EXAMPLE 248 (71?) -7- [ (Z) -2- (aminothiazol-4-yl) -2- (cyclopentyloxyimino)ace tamido] -3- (2-dimethylammoethylthiomethylphenylthio) -3-cephe m-4-carboxylate, inner salt
EXAMPLE 249 (71?) -7- [(Z) -2- (aminothiazol-4-yl) -2- (hydroxyimino) acetamido]
-3- [2,4-bis (amidinomethylthiomethyl)phenylthio] -3-cephem-4-carboxylate, trifluoroacetate salt
Thus, it will be appreciated that the compounds, methods and compositions of the invention are effective against various β-lactam resistant strains of bacteria which pose an increasing health risk to society. Although certain embodiments and examples have been used to describe the present invention, it will be apparent to those skilled in the art that changes to the embodiments and examples shown may be made without departing from the scope or spirit of the invention. Those references not previously incorporated herein by reference, including both patent and non-patent references, are expressly incorporated herein by reference for all purposes.
Other embodiments are within the following claims.

Claims

WHAT IS CLAIMED
A compound of the formula:
Figure imgf000150_0001
and its pharmaceutically acceptable salts and prodrugs, wherein:
R1 is selected from the group consisting of
-NHC(0)ZR3, -NRR5' and
Figure imgf000150_0002
Z is selected from the group consisting of -CH2(X)ra-, -C(NOR6)-, -CH(OR7)-, -C(CHC02R8)- and -CH(NR9R10) -;
X is selected from the group consisting of O and S;
m is is selected from the group consisting of 0 and 1;
R3 is selected from the group consisting of cyano, alkyl, aryl, heterocycle, heteroaralkyl and (CH2)nT,
n is 1 to 6,
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino;
R4"7 are independently selected from the group consisting of hydrogen, alkyl, aryl and acyl;
R8 is selected from the group of hydrogen, alkyl and aryl;
R9 and R10 are selected independently from the group consisting of hydrogen, alkyl, acyl, and heterocyclecarbonyl;
R2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl;
R11 is selected from the group consisting of H, halogen, alkyl, hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroaryliumalkyl; p is 0 , 1 , or 2 q is 0 or 1 r is 1 , 2 or 3
R12 is NR13R14 ,
Figure imgf000152_0001
Figure imgf000152_0002
R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring; and
R17 is H or alkyl.
2. The compound of claim 1, wherein R1 is
Figure imgf000152_0003
3. The compound of claim 2, wherein R3 is selected from the group consisting of alkyl, cyano, aryl, heterocycle, and (CH2)nT, n is 1 to 6 , and
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino.
4. The compound of claim 2, wherein R3 is selected from the group consisting of methyl, cyano, phenyl, thienyl, furanyl, 2-aminothiazolyl, and 2-aminothiadiazol-4-yl.
5. The compound of claim 2 wherein Z is -CH2(X)m-, and X is S.
6. The compound of claim 5 wherein m is 1 and R3 is alkyl or (CH2)nT.
7. The compound of claim 6, wherein Z is -C(NOR6)-, R6 is selected from the group consisting of hydrogen and alkyl, and R3 is heterocycle.
8. The compound of claim 7, wherein R6 is selected from the group consisting of hydrogen, methyl, 2-fluoroethyl, cyclopropylmethyl, allyl, dichloroallyl and cyclopentyl, and R3 is selected from the group consisting of phenyl, 2-thienyl, 2-furyl, 2-aminothiazol-4-yl, and 2-aminothiadiazol-4-yl .
9. The compound of claim 1, wherein R11 is selected from the group consisting of hydrogen, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl.
10. The compound of claim 1, wherein R11 is selected from the group consisting of hydrogen, trimethylammoniummethyl, N-alkylimidazoliummethyl, and pyridiniummethyl .
11. The compound of claim 1, wherein R11 is hydrogen.
12. The compound of claim 1, wherein there are two (CH2)p(S)q(CH2)rR12 substituents and R13-R17 are independently selected from the group consisting of hydrogen and lower alkyl.
13. The compound of claim 12, wherein p is 0 or 1, q is 0 or 1, r is 1, 2 or 3, R12 is NR13R14,
Figure imgf000154_0001
R13-R17 are hydrogen .
14 . The compound of claim 13 wherein p is 1 , + NR13R14
q is 1 , r is 1 or 2 , R12 is NR13R14 , ■Y NR15R16
Figure imgf000155_0001
Figure imgf000155_0002
, and R13-R17 are hydrogen.
15. The compound of claim 14 wherein there are three R11 substituents and two (CH2)p (S)q(CH2) rR12 substituents .
16. The compound of claim 1 wherein there are one or two (CH2)p(S) q(CH2)rR12 substituents,
R1 is -NHC(0)ZR3,
Z is -CH2(X)m- or -C(N0R6)-,
X is S,
R6 is selected from the group consisting of hydrogen and alkyl; R3 is selected from the group consisting of alkyl, cyano, aryl, heterocycle, and (CH2)nT,
R11 is selected from the group consisting of hydrogen, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl; and
R13-R17 are hydrogen or lower alkyl.
17. The compound of claim 16 wherein,
R11 is selected from the group consisting of hydrogen, trimethylammoniummethyl, N-alkylimidazoliummethyl, and pyridiniummethyl.
18. The compound of claim 17 wherein p is 0 or 1, q is 0 or 1, r is 1, 2 or 3, R12 is NR13R14,
Figure imgf000156_0001
R13-R17 are hydrogen .
19 . The compound of claim 18 wherein p is 1 , q is 1 , r is 1 or 2 , R12 is NR13R14 ,
Figure imgf000156_0002
, and Ri3 _Ri are hydrogen.
20. The compound of claim 17 wherein there are two (CH2)PR12 substituents,
R11 is hydrogen;
NR13R14
p is 1 , q is 1 , r is 1 or 2 , R12 is NR13R14 , ■V NR15R16
Figure imgf000157_0001
, and R13-R17 are hydrogen.
21. The compound of claim 1 wherein said pharmaceutically acceptable salt is an inorganic salt.
22. The compound of claim 21 wherein said inorganic salt is selected from the group consisting of chloride, bromide, iodide, nitrate, phosphate, or sulfate.
23. The compound of claim 1 wherein said pharmaceutically acceptable salt is a carboxylate salt.
24. The compound of claim 23 wherein said carboxylate salt is selected from the group consisting of acetate, propionate, butyrate, maleate, or fumarate.
25. The compound of claim 1 wherein said pharmaceutically acceptable salt is an alkylsulfonate.
26. The compound of claim 25 wherein said alkylsulfonate is selected from the group consisting of . methanesulfonate, ethanesulfonate, 2-hydroxyethylsulfonate, n-propylsulfonate, and iso-propylsulfonate.
27. The compound of claim 1 wherein said pharmaceutically acceptable salt is a hydroxycarboxylate.
28. The compound of claim 27 wherein said hydroxycarboxylate is selected from the group consisting of lactate, amlate, or citrate.
29. A method of treating a mammal suffering from a β-lactam resistant bacterial infection, comprising administering to such mammal a therapeutically effective amount of a compound of claim 1.
30. The method of claim 29, wherein said animal is infected with a β-lactam resistant
Staphylococcal or Enterococcal organism.
31. An antibacterial composition for treating a β-lactam antibiotic resistant bacterial infection, comprising a therapeutically effective amount of a compound of claim 1 in a pharmaceutically acceptable carrier.
32. The composition of claim 33, wherein said bacteria is a β-lactam resistant Staphylococcal or Enterococcal organism.
33. A method for using a thiolate nucleophile of the structure
Figure imgf000159_0001
wherein: R11 is selected from the group consisting of H, halogen, alkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl;
p is 0, 1 or 2
Y is OH, (S)q(CH2)rOH or (S)q(CH2) rR18 q is 0 or 1 r is 1, 2, or 3 and R18 is an optionally-protected version of one of the groups R12 , namely, NR13R14 ,
Figure imgf000160_0001
Figure imgf000160_0002
as a chemical reagent comprising the step of exposing said thiolate nucleophile to a cephem triflate of structure
Figure imgf000160_0003
or functionally equivalent intermediate wherein
R1 is selected from the group consisting of -NHC(0)ZR3,
Figure imgf000160_0004
Z is selected from the group consisting of -CH2(X)m-, -C(NOR6)-, -CH(OR7)-, -C(CHC02R8)- and -CH(NR9R10) -;
X is selected from the group consisting of 0 and S ;
m is is selected from the group consisting of 0 and 1; and R2 is selected from the group consisting of 5 hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl.
34. The method of claim 33 wherein said functional equivalent has a structure wherein the Tf 10 group is replaced with a halogen.
35. A method of using a compound of the structure
15
Figure imgf000161_0001
wherein
R11 is selected from the group consisting of H, r 20 halogen, alkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary ι heteroarylium alkyl;
and p is 1 or 2; as a reagent comprising the step of conversion of the free hydroxyl group to a leaving group, followed by exposing the resulting compound to a second compound of the structure H(S) q(CH2) rR18, wherein q is 0 or 1 r is 1, 2, or 3 and R18 is an optionally-protected version of one of the
groups R12 , namely, NR13R14 ,
Figure imgf000162_0001
R 3 R17 ς NR 5R16
■XX NR14 r or +"!NR13R14
wherein R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring, and
R17 is hydrogen or lower alkyl.
36. A method of using a compound of the structure
Figure imgf000162_0002
wherein
R11 is selected from the group consisting of H, halogen, alkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl;
p is 0, 1 or 2 q is 0 or 1 and r is 1, 2, or 3; as a reagent comprising the step of conversion of the free hydroxyl group to a leaving group, followed by exposing the resulting compound to a second compound of the structure
Figure imgf000163_0001
wherein
R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring.
37. A compound of the formula:
Figure imgf000164_0001
and its pharmaceutically acceptable salts and prodrugs, wherein:
R1 is-NHC(0)ZR3,
Z is selected from the group consisting of -CH2(X)m-, -C(N0R6)-, -CH(0R7)-, -C(CHC02R8)- and -CH(NR9R10) -;
X is selected from the group consisting of 0 and S;
m is is selected from the group consisting of 0 and 1;
R3 is (CH2)nT,
n is 1 to 6,
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino; R6"7 are independently selected from the group consisting of hydrogen, alkyl, aryl and acyl;
R8 is selected from the group of hydrogen, alkyl and aryl;
R9 and R10 are selected independently from the group consisting of hydrogen, alkyl, acyl, and heterocyclecarbonyl;
R2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl;
R11 is selected from the group consisting of H, halogen, alkyl, hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroaryliumalkyl;
p is 0, 1, or 2 q is 0 or 1 r is 1, 2 or 3
R« is NR13R14 ,
Figure imgf000165_0001
,
Figure imgf000166_0001
R13-R15 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring; and
R17 is H or alkyl.
38. The compound of claim 37, wherein R11 is selected from the group consisting of hydrogen, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl.
39. The compound of claim 37, wherein R11 is selected from the group consisting of hydrogen, trimethylammoniummethyl, N-alkylimidazoliummethyl, and pyridiniummethyl .
40. The compound of claim 37, wherein R11 is hydrogen.
41. The compound of claim 37, wherein R13-R17 are independently selected from the group consisting of hydrogen and lower alkyl.
42. The compound of claim 41, wherein p is 0 or 1, q is 0 or 1, r is 1, 2 or 3 , R12 is NR13R14,
Figure imgf000167_0001
R13-R17 are hydrogen.
43. The compound of claim 42 wherein p is 1,
NR13R14
q is 1, r is 1 or 2, R12 is NR13R14, Y NR15R16
Figure imgf000167_0002
Figure imgf000167_0003
, and R13-R17 are hydrogen.
44. A compound of the formula:
Figure imgf000167_0004
and its pharmaceutically acceptable salts and prodrugs, wherein:
R1 is selected from the group consisting of
-NHC(0)ZR3, -NR4R5 ' and
Figure imgf000168_0001
Z is selected from the group consisting of -CH2(X)m-, -C(NOR6)-, -CH(OR7)-, -C(CHC02R8)- and -CH(NR9R10) -;
X is selected from the group consisting of O and S;
m is is selected from the group consisting of 0 and 1;
R3 is selected from the group consisting of cyano, alkyl, aryl, heterocycle, heteroaralkyl and (CH2)nT,
n is 1 to 6,
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino; R4"7 are independently selected from the group consisting of hydrogen, alkyl, aryl and acyl;
R8 is selected from the group of hydrogen, alkyl and aryl;
R9 and R10 are selected independently from the group consisting of hydrogen, alkyl, acyl, and heterocyclecarbonyl;
R2 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl;
R11 is selected from the group consisting of H, halogen, alkyl, hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroaryliumalkyl;
p is 0 , 1 , or 2 q is 1 r is 1 , 2 or 3
R12 is NR13R" ,
Figure imgf000169_0001
Figure imgf000170_0001
R13-R16 are independently selected from the group consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl, and phosphoryl and taken together may form a 5- or 6-membered ring; and
R17 is H or alkyl.
45. The compound of claim 44, wherein R1 is
Figure imgf000170_0002
46. The compound of claim 45, wherein R3 is selected from the group consisting of alkyl, cyano, aryl, heterocycle, and (CH2)nT,
n is 1 to 6, and
T is selected from the group consisting of amino, amidino (C- or N-linked) , guanidino, and isothioureido, optionally substituted by alkyl, aryl, hydroxyl, or amino.
47. The compound of claim 45, wherein R3 is selected from the group consisting of methyl, cyano, phenyl, thienyl, furanyl, 2-aminothiazolyl, and 2-aminothiadiazol-4-yl .
48. The compound of claim 45 wherein Z is -CH2(X)m-, and X is S.
49. The compound of claim 45 wherein m is 1 and R3 is alkyl or (CH2)nT.
50. The compound of claim 49, wherein Z is -C(N0R6)-, R6 is selected from the group consisting of hydrogen and alkyl, and R3 is heterocycle.
51. The compound of claim 50, wherein R6 is selected from the group consisting of hydrogen, methyl, 2-fluoroethyl, cyclopropylmethyl, allyl, dichloroallyl and cyclopentyl, and R3 is selected from the group consisting of phenyl, 2-thienyl, 2-furyl, 2-aminothiazol-4-yl, and 2-aminothiadiazol-4-yl.
52. The compound of claim 44, wherein R11 is selected from the group consisting of hydrogen, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl.
53. The compound of claim 44, wherein R11 is selected from the group consisting of hydrogen, trimethylammoniummethyl, N-alkylimidazoliummethyl, and pyridiniummethyl .
54. The compound of claim 44, wherein R11 is hydroge .
55. The compound of claim 44, wherein R13-R17 are independently selected from the group consisting of hydrogen and lower alkyl .
56. The compound of claim 55, wherein p is 0
+ NR13R14
or 1, q is 1, r is 1, 2 or 3 , R12 is NR13R14, ,
R3 NR15R16 ς R15R16
YY NR14 ,
Figure imgf000172_0001
f or *V +ΪNR13R14 , and R13-R17 are
hydrogen .
57. The compound of claim 56 wherein p is 1,
q is 1, r is 1 or 2, R12 is NR13R14,
Figure imgf000172_0002
,
Figure imgf000172_0003
, or R3 17 N /R"
\.
NR ,14 and R13-R17 are hydrogen.
58. The compound of claim 1 wherein
R1 is -NHC(0)ZR3, Z is -CH2(X)m- or -C(NOR6)-,
X is S,
R6 is selected from the group consisting of hydrogen and alkyl;
R3 is selected from the group consisting of alkyl, cyano, aryl, heterocycle, and (CH2)nT,
R11 is selected from the group consisting of hydrogen, optionally substituted aminoalkyl or quaternary ammonium alkyl, and quaternary heteroarylium alkyl; and
R13-R17 are hydrogen or lower alkyl.
59. The compound of claim 58 wherein,
R11 is selected from the group consisting of hydrogen, trimethylammoniummethyl, N-alkylimidazoliummethyl, and pyridiniummethyl.
60. The compound of claim 59 wherein p is 0 or 1, q is 1, r is 1, 2 or 3 , R12 is NR13R14,
Figure imgf000174_0001
R13-R17 are hydrogen.
61. The compound of claim 60 wherein p is 1, q is 1, r is 1 or 2, R12 is NR13R14,
Figure imgf000174_0002
, and , R .1,3-R .1_7 are
hydrogen.
62. The compound of claim 17 wherein
R11 is hydrogen;
Figure imgf000174_0003
Figure imgf000174_0004
t and R13-R17 are hydrogen.
PCT/US1996/008928 1995-05-31 1996-05-29 Cephalosporin antibiotics WO1996038450A1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001021623A1 (en) * 1999-09-22 2001-03-29 Essential Therapeutics, Inc. 7-acylamino-3-heteroarylthio-3-cephem carboxylic acid antibiotics and prodrugs thereof

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6271222B1 (en) * 1998-05-28 2001-08-07 Merck & Co., Inc. Penem antibacterial compounds, compositions and methods of treatment
HUP0500956A3 (en) * 2000-10-19 2013-01-28 Amura Ltd Oxapenem-3-carboxylic acids and pharmaceutical compositions containing them
EP1199077A1 (en) * 2000-10-19 2002-04-24 Amura Limited Stable compositions of oxapenem-3-carboxylic acids by Co-lyophilisation with pharmaceutical carriers
CN112321606B (en) * 2020-11-04 2022-05-17 福安药业集团重庆博圣制药有限公司 Preparation method of ceftizoxime sodium and intermediate thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3992377A (en) * 1974-12-13 1976-11-16 Eli Lilly And Company 3-Thio-substituted cephalosporin antibiotics
CH605998A5 (en) * 1974-08-30 1978-10-13 Ciba Geigy Ag 3-Thio-7-amino-3- and-2-cephem-4-carboxylic acids
CH605999A5 (en) * 1974-08-30 1978-10-13 Ciba Geigy Ag 3-Thio-7-amino-3- and-2-cephem-4-carboxylic acids
US4123528A (en) * 1975-11-21 1978-10-31 Merck & Co., Inc. 3-(Substituted thio) cephalosporins, derivatives and nuclear analogues thereof
US5162521A (en) * 1991-06-06 1992-11-10 Bristol-Myers Squibb Company Processes for making cephems from allenylazetidinone derivatives
WO1995026966A1 (en) * 1994-04-01 1995-10-12 Microcide Pharmaceuticals, Inc. Cephalosporin antibiotics

Family Cites Families (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS609515B2 (en) * 1976-08-09 1985-03-11 塩野義製薬株式会社 3'-Noroxacephalosporins
US4168314A (en) * 1977-11-17 1979-09-18 Merck & Co., Inc. 6-(1'-Hydroxyethyl)-2-aminoethylthio-pen-2-em-3-carboxylic acid
DK446179A (en) * 1978-10-24 1980-05-29 Merck & Co Inc PROCEDURE FOR PREPARING SUBSTITUTED 1-CARBADETHYAPE-2-EM-3-CARBOXYL ACIDS
DE2965394D1 (en) * 1978-11-01 1983-06-16 Sanraku Ocean Co Process for producing antibiotic beta-lactam compounds
EP0074599A1 (en) * 1981-09-09 1983-03-23 Takeda Chemical Industries, Ltd. 5,6-cis-Carbapenem derivatives, their production and use
US4640915A (en) * 1982-03-29 1987-02-03 Fujisawa Pharmaceutical Co., Ltd. 1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid derivatives
EP0113101A1 (en) * 1982-12-30 1984-07-11 Merck & Co. Inc. 6-(1-Hydroxyethyl)-2-SR8-1-methyl-1-carbadethiapen-2-em-3-carboxylic acid esters
JPS60174787A (en) * 1984-02-21 1985-09-09 Kyowa Hakko Kogyo Co Ltd 3-position-substituted carbacephem compound
JPS60202886A (en) * 1984-03-27 1985-10-14 Sankyo Co Ltd 1-substituted carbapenem-3-carboxylic acid derivative and its preparation
US4806637A (en) * 1986-03-17 1989-02-21 Schering Corporation 6-(Hydroxyethyl)-2-(heterocyclylthio)-penem-3-carboxylates
EP0243686B1 (en) * 1986-03-27 1992-07-15 Sumitomo Pharmaceuticals Company, Limited Beta-lactam compounds, and their production
DE3920743A1 (en) * 1989-06-24 1991-01-10 Bayer Ag 2-ISOCEPHEME AND 2-OXA-ISOCEPHEME, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE AS AND IN MEDICINAL PRODUCTS
AU636913B2 (en) * 1989-10-11 1993-05-13 Takeda Chemical Industries Ltd. Tricyclic carbapenem compounds
US5077287A (en) * 1991-01-18 1991-12-31 Eli Lilly And Company 3-thiazolylthio carbacephem antibacterial agents
GB9104838D0 (en) * 1991-03-07 1991-04-17 Glaxo Spa Heterocyclic compounds
GB9104769D0 (en) * 1991-03-07 1991-04-17 Glaxo Spa Heterocyclic compounds
EP0507313A1 (en) * 1991-04-05 1992-10-07 Takeda Chemical Industries, Ltd. Polycyclic carbapenem compounds, their production and use
US5587474A (en) * 1992-06-18 1996-12-24 Tanabe Seiyaku Co., Ltd. Method for removing the protecting group for carboxyl group

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH605998A5 (en) * 1974-08-30 1978-10-13 Ciba Geigy Ag 3-Thio-7-amino-3- and-2-cephem-4-carboxylic acids
CH605999A5 (en) * 1974-08-30 1978-10-13 Ciba Geigy Ag 3-Thio-7-amino-3- and-2-cephem-4-carboxylic acids
US3992377A (en) * 1974-12-13 1976-11-16 Eli Lilly And Company 3-Thio-substituted cephalosporin antibiotics
US4123528A (en) * 1975-11-21 1978-10-31 Merck & Co., Inc. 3-(Substituted thio) cephalosporins, derivatives and nuclear analogues thereof
US5162521A (en) * 1991-06-06 1992-11-10 Bristol-Myers Squibb Company Processes for making cephems from allenylazetidinone derivatives
WO1995026966A1 (en) * 1994-04-01 1995-10-12 Microcide Pharmaceuticals, Inc. Cephalosporin antibiotics

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001021623A1 (en) * 1999-09-22 2001-03-29 Essential Therapeutics, Inc. 7-acylamino-3-heteroarylthio-3-cephem carboxylic acid antibiotics and prodrugs thereof
RU2225868C2 (en) * 1999-09-22 2004-03-20 Исеншл Терапьютикс, Инк. Derivatives of 7-acylamino-3-heteroarylthio-3-cephem carboxylic acid and antibacterial compositions comprising thereof

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