[go: up one dir, main page]

US3316273A - Penicillin aldehydes - Google Patents

Penicillin aldehydes Download PDF

Info

Publication number
US3316273A
US3316273A US353577A US35357764A US3316273A US 3316273 A US3316273 A US 3316273A US 353577 A US353577 A US 353577A US 35357764 A US35357764 A US 35357764A US 3316273 A US3316273 A US 3316273A
Authority
US
United States
Prior art keywords
acid
thiopenicillanic
penicillanal
acetamido
solution
Prior art date
Legal status (The legal status 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 status listed.)
Expired - Lifetime
Application number
US353577A
Inventor
William J Gottstein
Lee C Cheney
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Bristol Myers Co
Original Assignee
Bristol Myers Co
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 to BE661405D priority Critical patent/BE661405A/xx
Application filed by Bristol Myers Co filed Critical Bristol Myers Co
Priority to US353577A priority patent/US3316273A/en
Priority to DE19651545581 priority patent/DE1545581A1/en
Priority to FR10033A priority patent/FR1479231A/en
Priority to CH385065A priority patent/CH527846A/en
Priority to GB11824/65A priority patent/GB1097338A/en
Priority to NL6503503A priority patent/NL6503503A/xx
Priority to AT261265A priority patent/AT267055B/en
Application granted granted Critical
Publication of US3316273A publication Critical patent/US3316273A/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • 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

Definitions

  • Preferred compounds of the present invention are those of the formulae PENICILLlN ALDEHYDES William J. Gottstein and Lee c. Cheney, Fayetteville,
  • This invention relates to novel antibacterial agents and f chemical reagents and, more particularly, to 6-substituted ArXC-C-NHOHOH C CH3 H penicillanyl aldehydes and especially to penicillin alde- LW L Q hydes.
  • the aldehydes of the present invention are effective antibacterial agents in vivo against Gram-positive bacteria 3 and are also effective agents for use in a search for oXida- 15 Z H tive enzymes.
  • penicillins include those disclosed in US. Patents 2,941,995, 2,951,839, 2,985,648, 2,996,601, 3,007,- 920, 3,025,290, 3,028,379, 3,035,047, 3,040,032, 3,040,033, 3,041,332, 3,041,333, 3,043,831, 3,053,831, 3,071,575, (C H CNHOH-CH 0-011 H 3,071,576, 3,079,305, 3,079,306, 3,080,356, 3,082,204, I, 3,093,547, 3,093,633, 3,117,119, 3,118,877, 3,120,512, 3,120,513, 3,120,514; in British patent specifications 6 874,414, 874,416, 876,516, 876,662, 877,120, 877,323, C 877,531, 878,233, 880
  • R represents hydrogen, amino, carbobenzoxymino, phenyl, fluoro, chloro, bromo, iodo, hydroxy, or lower)alkanoyloxy including especially acetoxy or (lowr)alkoxy;
  • X represents oxygen or sulfur;
  • R and R each epresent hydrogen, phenyl, benzyl, phenet-hyl or (lowr)alkyl;
  • R represents (lower)alkyl;
  • R and R each repesent (lower)alkyl, (lower)alkylthio, benzylthio, cyclo- 1 II Ar-C- and Ar represents the monovalent radical of the formula wherein R R and R are each a member selected from the group consisting of hydrogen, chloro, bromo
  • (lower)alkyl as used herein means both straight and branched chain aliphatic hydrocarbon radicals having from one to ten carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl, 2-ethylhexyl, heptyl, decyl, etc.
  • (l0wer) alkoxy it refers to the alkyl portion of such group which is therefore as described above in connection with (lower)alkyl.
  • Preferred embodiments of the present invention are the restricted series of compounds of the following formulae wherein R represents (lower) alkyl
  • R represents (lower)alkyl and R and R each represent a member selected from the group consisting of hydrogen and chloro;
  • R is (lower)alkyl and R is a member selected I, from the group consisting of hydrogen and chloro; 0
  • the compounds of the present invention are prepared,- according to the present invention, by either of two methods, as follows:
  • the present penicillin aldehydes are prepared by oxidation of the corresponding penicillin alcohols by the addition of about 2-5 moles of a carbodiimide (e.g. dicyclohexylcarbodiimide, diisopropylcarbodiimide) to a solution of about one mole of the alcohol and 0.1-2.0 mole anhydrous phosphoric acid in dry dimethyl sulfoxide.
  • a carbodiimide e.g. dicyclohexylcarbodiimide, diisopropylcarbodiimide
  • Tetramethylene sulfoxide may also be used and the sulfoxide may be diluted with up to nine volumes of an inert solvent.
  • the reaction proceeds readily at room temperature.
  • the phosphoric acid functions as a catalyst and may be replaced with phosphorous acid, cyanoacetic acid or pyridinium phosphate, trifluoroacetate, hydrochloride or sulfate. Further details are given in the
  • the penicillin aldehydes of the present invention are preferably prepared by hydrogenolysis by Raney nickel of a thiopenicillin (also called a penicillin thioacid; e.g. see US. Patent 2,751,378) which is preferably in the form of the free acid or of a salt such as an alkali metal salt or an amine salt.
  • the reaction is carried out at 50 C. and preferably at close to 0 C.
  • anhydrous or nearly anhydrous solvent such as tetrahydrofuran or acetone and to use catalytic amounts of a weak anhydrous acid such as glacial acetic acid as the penicillin aldehydes are very sensitive to bases.
  • a weak anhydrous acid such as glacial acetic acid
  • the proportions are not critical but it is of course desirable to use enough Raney nickel to assure a high yield in the reaction; it is thus preferred to use a weight of Raney nickel (on a Wet basis) equal to at least three times the weight of the thiopenicillin.
  • the activated Raney nickel available commercially is quite suitable for use as is that prepared according to Mozingo et al., J. Amer. Chem. Soc., 65, 1013 (1943).
  • the thiopenicillins can be replaced, if desired, with their esters, e.g. with alkylthio or aralkylthio esters; see J. Amer. Chem. Soc., 75, 3636-3637 (1953), J. Chem. Soc. (London), 3733-3739 (1953) and Chem. Ber., 92, 530-534 (1959).
  • the penicillin aldehyde can be readily regenerated by treatment of these heterocyclic derivatives with an acid, e.g., with p-toluenesulfonic acid monohydrate in a suitable solvent such as acetone-ether.
  • Examples of preferred diamines for use as the aldehyde trapping agents are those of US. Patent 2,739,981 and especially N,N'-dibenzylethylenediamine [W. F. Minor, D. A. Johnson and L. C. Cheney, J. Org. Chem., 21, 528 (1956)], N,N-diphenylethylenediamine [W. Wanzlick and W. Lochel, Chem. Ber., 86, 1463 (1953)].
  • Other suitable diamines include N,N'-dimethy1ethylenediamine [A. J. Birch, J. Cymerman Craig and M. Slaytor, Australian J.
  • the product is easily separated from the Raney nickel by filtration and from unreacted starting thiopenicillin by virtue of the fact that only the latter contains an acidic group and can thus be extracted from an organic solvent, such as ether, into aqueous alkali, e.g. 5% NaI-ICO
  • the penicillin aldehydes may be purified by reaction with 2,4-dinitrophenylhydrazine or N,N-di'benzylethylenediarnine [of J. Org. Chem, 21, 528-529 (1956) and US.
  • the compounds of the present invention exhibit in vitro antibacterial activity.
  • MIC Minimum Inhibitory Concentration
  • heart infusion broth to which 5% pooled human serum had been added
  • t was surprisingly discovered that these compounds are :ffective antibacterial agents in vivo, that is, exhibited a ninirnum curative dose in 50% of the mice tested (CD against an overwhelmingly lethal dose of S. aureus Smith is follows:
  • EXAMPLE 1 O C H3 6-phenoxyacetamidopenicillanal.
  • Commercial, pyrophoric Raney active nickel catalyst 125 g. wet weight, was washed three times with 275 ml. portions of absolute ethanol and then with four 275 ml. portions of tetrahydrofuran (Tl-IF).
  • the nickel was then suspended in 500 ml. of THF, 10.5 ml. (0.175 mole) of glacial acetic acid was added and the mixture was stirred and cooled to 4.
  • a solution of 20.22 g. (0.0500 mole) of the potassium salt of penicillin V thiol acid was prepared by suspending the solid in 200 ml. THF and adding 15 ml. Water.
  • the compounds of the present invention are useful agents for the detection of microorganisms containing oxidative enzymes, e.g. of the type used to oxidize steroids.
  • oxidative enzymes e.g. of the type used to oxidize steroids.
  • cells of the microorganisms being investigated are grown for about 24 hours in a suitable medium (e.g. heart infusion broth with glucose, yeast-malt medium) on a shaker (e.g. at 28 or 37 C.).
  • the cells from 10 ml. are obtained by centrifugation and added to 2 ml. of substrate solution at each of various pHs such as 5 and 7, e.g. to a solution of 500 mcg./ ml.
  • phenoxymethylpenicillin aldehyde (IV) in 20% acetone-80% 0.2 molar pH 5 citrate bufier That mixture of cells and substrate is then returned to the shaker for a short period of time (e.g. four hours). Aliquots are taken, diluted (e.g. ten-, fiftyand one hundred-fold) and assayed by a typical penicillin assay, e.g.
  • DMF Dimethylformamide
  • Ethyl chloroformate 9.38 ml. (0.0985 mole)
  • the mixture was stirred and cooled down to 5 during the next ten minutes forming the mixed anhydride.
  • a solution of 2 2 g. (0.2 mole) of sodium hydrosulfide (hydrated) in 350 ml. of DMF was prepared by stirring for ten minutes at room temperature under a nitrogen atmosphere. The solution was a turbid yellow, but all chunks of the salt had B. subtilis; activity in this assay indicates the presence of disappeared at this time.
  • the solution of sodium hydro- .9 sulfide was poured into the mixed anhydride solution, and the resulting dark mixture was stirred at +l to 0 for ten minutes forming the thio acid.
  • the reaction mixture was immediately poured into 3700 ml. of ice water, acidified to pH 2 with ca. 50 ml. of 6 N H 80 and extracted with two 1500 ml. portions of ice-cold ether.
  • the combined yellow ether extracts were washed twice with 75 ml. portions of ice water, and then extracted with two 200 ml. portions of 4.25% NaHCO solution which carried the yellow color into the aqueous layer.
  • the combined basic extracts were added to a solution of 36 g.
  • the nickel was then suspended in 100 ml. of THF and 2.1 ml. (0.035 mole) of glacial acetic acid and 12 g. (0.0050 mole) of N,N'-dibenzylethylenediamine (DBED) was added and the suspension was stirred and cooled to A cooled (5) solution of 4.86 g. (0.0100 mole) of pen V thiol DBED salt in 40 ml. of THF was added and the mixture was stirred at 2' for hour. The yield and quality of product may possibly be substantially improved by reducing this reaction time to about 15 minutes. The mixture was filtered over diatomaceous earth (Supercel) and the catalyst was washed with four 30 ml. portions of THF.
  • Supercel diatomaceous earth
  • the THF was removed under vacuum at 33 and the dark residue was dissolved in 300 ml. of ether. A considerable amount of gum failed to dissolve. On addition of 50 ml. of water, part of the gum dissolved. In some runs a pink solid separated at this point and was filtered off. The water layer was separated and the ether solution was extracted successively with two 30 ml. portions of 5% acetic acid, 50 ml. of water, three 30 ml. portions of 5% sodium bicarbonate and three 30 ml. portions of Water. The ether solution was dried briefly over sodium sulfate, treated with decolorizing carbon and filtered over Supercel which gave a nearly colorless filtrate.
  • the solvent was flashed ofl at 33, the residue was flashed with two 100 ml. portions of chloroform to remove moisture and then thoroughly dried over phosphorus pentoxide at 0.5 mm.
  • the yield of the titled compound was 2.23 g. of a hard gum.
  • EXAMPLE 3 s CH3 6-(3-phenyl-5-meflzylfsoxazole 4 carboxamido)penicillamal.-One hundred grams of wet No. 28 commercial Raney Ni (Raney Co.) was washed three times with ml. portions of absolute ethanol, three times with 100 ml. portions of acetone and finally a fourth portion of 100 ml. of acetone was added and the slurry cooled and stirred at 0 to 5 C. while 7 g. of N,N-diphenylethylenediamine, 2.4 ml. of glacial acetic acid and a solution of 10 g.
  • Raney Ni Raney Co.
  • Penicillin G aldehyde (6 phenylacetamidopeniciL lanal).-To a precooled, stirred suspension of 13.08 g. (0.03 mole) of penicillin G-triethylammonium salt in 200 ml. of dimethylformamide (DMF) was added 3.24 g. (0.03 mole) of ethyl chloroformate dropwise over a ZOminute period. The resulting nearly clear solution was then treated all at once with 3 g. of NaSH in 50 ml. of DMF which had been previously dissolved by pulverizing in a mortar and stirred and cooled in an ice bath until a cloudy solution had been obtained.
  • DMF dimethylformamide
  • the resulting dark green reaction mixture was stirred one-half hour with the ice bath removed and poured into a mixture of 500 ml. benzene, one liter of crushed ice in water and 30 ml. of 40% H PO
  • the benzene extract was washed three times with cold water and partially dried by filtering through sodium sulfate. The benzene was then removed under reduced pressure on the flash evaporator and when the benzene was nearly all removed (avoiding heating) the resulting oil was dissolved in 50 ml. of dry acetone and added immediately, dropwise, to a previously prepared stirred suspension of 100 g. of No.
  • the ethereal filtrate was washed three times with ml. portions of water, three times with 100 ml. portions of 2% NaHCO solution and three times with water and dried over Na SO
  • a solution of p-toluene sulfonic acid (5 g. in 50 ml. acetone) until the solution was acidic to wet pH paper.
  • the solid p-toluenesulfonic acid salt of N,N-diphenylethylenediamine which precipitated was removed by filtration and the filtrate was treated as before with the acid repeatedly until no more solid formed upon addition of acid and the ethereal solution was still acidic to wet filter paper after standing ten minutes.
  • Benzyloxypenicillin aldehyde (6 N carbobenzyloxyaminopenicillanaL-To 45 g. of previously acetonewashed Raney nickel suspended in 100 ml. of acetone with 3.7 g. (0.0174 mole) N,N'-diphenylethylenediamine and 2.1 m1. of glacial acetic acid, was added 7 g. (0.0174 mole) of benzyloxypenicillin thiol (potassium salt) dissolved in 25 ml. of acetone and 3 ml. of glacial acetic acid. The mixture was stirred for one hour in an icebath and the mixture was filtered.
  • the filtrate was evaporated to an oil under reduced pressure at 30 and residue was sl-urried with 100 ml. of ether.
  • the ether was washed twice with dilute sodium bicarbonate and several times with water.
  • the ether was dried over anhydrous magnesium sulfate.
  • the drying agent was separated and the ether solution treated with a solution of p-toluenesulfonic acid in acetone until there was no more turbidity.
  • the N,N'-diphenylethylenediamine toluenesulfonate was collected and the filtrate was washed with water and dried over anhydrous MgSO
  • the solvent was evaporated under reduced pressure to yield 1.5 g. of neutral benzyloxypenicillin aldehyde.
  • EXAMPLE 7 6 phthalimidopenicfllin aldehyde (6 phthalimidopenicillanal).T0 a solution of 5.0 g. (0.015 mole) of 6-phthalimidopenicillanyl alcohol in 50 ml. of anhydrous dimethylsulfoxide was added 10.0 g. (0.049 mole) of dicyclohexylcarbodiimide and 1.5 g. (0.0075 mole) of pyridinium trifiuoracetate. The mixture was swirled in an Erlenmeyer flask for a few seconds, placed in a desiccator under a nitrogen atmosphere, allowed to stand at 25 C. for 17 hours and then filtered and the filtrate thrown into 500 ml.
  • the nickel was then suspended in 100 ml. THF and cooled to 4 and to this there was added 2.1 ml. glacial acetic acid. Potassium 6-(5-methyl-3-o-chlorophenylisoxazole-4-carboxamido)thiolpenicillanate (4.5 g.) was dissolved completely in 40 m1. THF and added to the Raney nickel. The reaction mixture was stirred below 0 for 15 minutes. The Raney nickel was then filtered through diatomaceous earth and washed five times with 60 ml.
  • the resulting oil was partially dissolved in 350 ml. ether. A pink gum appeared which was carried along with the ether. The ether was then extracted three times with 60 m1. portions of 5% NaHCO and three times with 40 ml. portions of water. The ether layer was then separated from the gum and dried over magnesium sulfate and charcoal. After filtration the ether was evaporated to dryness. The resulting oil could be solidified by dissolving it in methylene chloride or ether and precipitating with n-pentane.
  • EXAMPLE 9 In the procedure of Example 5 the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids 6-[a-(2-ch1orophenoxy)propionamido] thiopenicillanic acid, 6-[a-(4sultamylphenoxy)-n-butyramido] thiopenicillanic acid,
  • each of which is is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system.
  • EXAMPLE 1 1 In the procedure of Example 5 the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
  • each of which is isolated as a solid which s converted to an active antibacterial agent upon exposure an oxidative enzyme system.
  • EXAMPLE 16 In the procedure of Example 5, the potassium thioahenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
  • EXAMPLE 18 6-[a-(4-nitrophenyl) acetamido]thiopenicillanic acid, 6- cc- (4-bromopheny1 acetamido] thiopenicillanic acid, 6- [u-(4-t-butylphenyl) acetamido1thiopenicillanic acid,
  • 6- [o.-(3-diethylaminopheny1) acetamido1penicillanal, and 6- [oc- (2,4-diisoamylphenyl acetamido1pencillanal,
  • EXAMPLE 22 (1) 6-phenoxyacelamidopenicillanic thiol acid potassium salts.-A solution containing 35.04 g. (0.100 mole) of penicillin V (phenoxymethylpenicillin) 14.02 ml. (0.10 mole) of dry triethylamine, and 508 ml. of pure, dry DMF (dimethylformamide) was stirred and cooled to -3 C. Ethyl chloroforrnate (9.51 ml.; 0.10 mole) was added and the resulting solution was stirred at 0 for 10 minutes. A solution of 22.0 g. (0.2 mole) of sodium hydrosulfide (hydrated) in 358 ml. of pure, dry DMF was prepared by stirring at 25 C. under a nitrogen atmosphere for 10 minutes. This turbid yellow solution was added to the mixed anhydride solution all at once and the cooling bath was removed. After stirring for 10 minutes,
  • reaction mixture was poured into 3.0 liters of ice water.
  • the solution was acidified to pH 2.0 with 6 N sulfuric acid and extracted with three one-liter portions of cold ether.
  • the combined ether solutions were washed twice with 500 ml. portions of ice water, dried very briefly over sodium sulfate and filtered.
  • Addition of 20.5 ml. of an by weight solution of potassium-Z-ethylhexanoate (KEH) in dry n-butanol (diluted to 40 ml. with dry ether) precipitated the product, which was collected by filtration, Washed with dry ether and dried, yield 32.7 g. (81%).
  • the product was recrystallized by dissolving in a minimum of cold water (ca.
  • the filtrate was extracted with 20 ml. of 10% phosphoric acid, which caused N,'Ndiphenylethylenediarnine phosphate to crystallize. It was filtered off, the aqueous phase was separated, and the other solution was again extracted with 20 ml. of 10 phosphoric acid and washed with three 100 ml. portions of water. It was dried over sodium sulfate, filtered, flashed dry and flashed with ethyl acetate. The addition of 10 ml. of dry ether to the small residue caused 0.25 g. of product to crystallize, M.P. 164168 C.
  • the infrared absorption spectrum was quite sharp, howing NH 3380 c-m.- fl-lactam 1780, aldehyde 1740, mide 1670 and 1525 cmf All of the bands noted were -f equal intensity.
  • the aldehyde may be recrystallized cut at 33 C. Water (a few ml.) and ordinary ether cetate (three parts per 100 ml.) and flashing off the solent at 33 C. Water (a few ml.) and ordinary ether one part per 100 ml.) are added and the resulting soluion is allowed to stand at 20 C. for 24 hours. The prodict will slowly crystallize.
  • the infra-red spectrum (5% in CCl shows NH at 3400 cmr ,8- lactam 1800, enol acetate carbonyl 176 2, amide 1700 and 1510 and COC bands at 1230, 1190, and 1090 crnf A weak band at 1640 cm.- is ascribed to
  • the NMR spectrum is in full agreement with the enol acetate structure and shows further that the product is a mixture of approximately equal amounts of cis and trans isomers.
  • Another series of starting materials used to prepare the compounds of the present invention comprises the series of compounds of the formula wherein R is amino, acylarnino, ibenzyloxycarbonylamino, phthalimido or tritylamino and, particularly, the so-called penicillin thioacids or thiopenicillins of the formula S CH3 ll in which R represents the side chain of any of the known penicillins other than those containing a group which is altered by reaction with Raney nickel above 0 C.
  • These compounds are usually used in the form of salts, e.g. sodium, potassium.
  • thiopenicillins are prepared from penicillins by the methods described below or in US. Patent 2,751,378.
  • an active acylating derivative of the starting penicillin such as an anhydride or a mixed anhydride (such as the mixed anhydride with a lower alkyl ester of ethoxy-or isobutoxy-carbonic acid) or an acid chloride is prepared and reacted with a source of sulfhydryl groups, e.g. hydrogen sulfide or sodium hydrosulfide or potassium hydrosulfide.
  • the 6 (DL-oc phenoxypropionamido)thiopenicillanic acid is extracted into ether, washed with water, and dried over anhydrous magnesium sulfate. Potassium 2-ethy1- hexanoate (5 gm.) is added and the crystalline precipitate is collected and weighs 4.5 gm. Recrystallization from water and acetone yields 1.1 gm. of the product, potassium 6 (DL a phenoxypropionamido)thiopenicil lanat'e, as colorless plates which are found to contain the B-lactam ring as shown by infrared analysis, to have a melting point of greater than 240 C. with decomposition and the following elemental anaylsis:
  • potassium 6-(D'a-phenoxypropionamido)thiopenicillan-ate which is found to contain the fi-lactam ring as shown by infrared analysis, to have a melting point of greater than 195 C. with decomposition and the following elemental analysis:
  • Preparation 14 Preparation of potassium 6-(L-aphenoxypropionamido)thiopenicillanate
  • Dilute sulfuric acid is added to a solution of potassium 6 (L-u-phenoxypropionamido)penicillanate (5 gm., 0.0125 mole) in water ml.) until a pH of 2 is attained.
  • the penicillin acid is extracted from this solution into ethyl acetate (200 ml.), washed with water, and dried over anhydrous magnesium sulfate.
  • the ethyl acetate is evaporated at 35 C.
  • the residue is dissolved in dimethylformamide (150 ml.) and cooled to 5 C. in an ice bath.
  • 2,6-lutidine (1.33 gm., 0.0125 mole) is added, followed by the dropwlse addition of ethyl chloroformate (1.33 gm., 0.0125 mole).
  • the mixture is stirred for 15 minutes and'a suspension of sodium hydrosulfide (2.5 gm., 0.0447 mole) in dimethyl'formamide (100 ml.) is added all at once.
  • the solution is stirred for 20 minutes and then poured into water (one liter) and acidified to pH 2 with dilute sulfuric acid.
  • the 6-(L-a-phenoxypropionamido)thiopenicillanic acid is extracted into ether, washed with water, and dried over anhydrous magnesium sulfate.
  • Potassium 2-ethylhexan0ate (3 gm.) is added and the crystalline precipitate is collected. Recrystallization from water and n-butan-ol yields 2.5 gm. of the product, potassium 6-(L-ot-phenoxypropionamido)thiopenicillanate which is found to have a melting point of greater than 215 C. with decomposition and the following elemental analysis:
  • Preparation 15--Preparati0n of potassiumo-(aisopropyl-u-phenoxyacetamido)thiopenicillanate Dilute sulfuric acid is added to a solution of potassium 6-(a-isopropyl-a-phenoxyacetamido)thiopenicillanate (5.3 gm., 0.0125 mole) in water (150 ml.) until a pH of 2 is attained. The penicillin acid is extracted from this solution into ethyl acetate (200 m1.) washed with water, and dried over anhydrous magnesium sulfate. The ethyl acetate is evaporated at 35 C.
  • the 6-(asopropyl-a phenoxyacetamido)thiopenicillanic acid is ex- :racted into ether, Washed with water, and dried over anhydrous magnesium sulfate. Potassium Z-ethylhexanoate (3 gm.) is added and the crystalline precipitate is :ollected. Recrystallization from ethyl acetate and Skellysolve yields 2.3 gm. of the product potassium 6-(alsopropyl-a-phenoxyacetamido)thiopenicillanate, which is found to contain the B-lactam ring as shown 'by infrared analysis and to have a melting point of greater than 170 C. with decomposition.
  • Preparation 1 6 --Preparation of potassium 6-[L( -ocphenoxybutyram ido] thio p enici l lanate Potassium 6- [L( )-a-phenoxybutyramido]penicillanate (2.08 gm., 0.005 mole) is dissolved in water (30 ml.) and layered with ethyl acetate. After cooling to 5 C., dilute phosphoric acid (40%) is added until a pH of 2 is attained. The penicillin acid is extracted into the ethyl acetate and a further extraction with fresh ethyl acetate is made.
  • the resulting green solution is stirred for 25 minutes and poured into a precooled (10 C.) mixture of water (150 ml.), acidified to pH 1.5 with dilute phosphoric acid (40%) and benzene (100 ml.) with vigorous stirring.
  • the 6- [L()-a-phenoxybutyramido]thiopenicillanic acid is extracted into benzene and a further extraction with fresh benzene is made.
  • the combined extracts are washed and dried. Potassium 2-ethylhexanoate (0.005 mole) is added as a 50% solution of potassium Z-ethylhexanoate in butanol.
  • Skellysolve B (a petroleum ether fraction having a boiling point range of from about 60 to about 68 C. consisting essentially of n-hexane) is added to the solution with cooling and shaking until the solution becomes cloudy, and an oil separates. The solution is decanted and flashed to dryness. The residue is triturated with ether, and a white solid forms which is removed by filtration, washed with dry ether, and dried in vacuo. The product, potassium 6- [L( )-a-phenoxybutyramido] thiopenicillanate is found to weigh 2 grams, to have a melting point of 145 149 C. with decomposition and the structure is confirmed by infrared analysis.
  • Preparation 18 In the procedure of Preparation 11, the potassium 6- (a-phenoxypropionamido)penicillanic acid is replaced by 0.025 mole of the potassium salt of 6- a-phenylthiopropionamido penicillanic acid, 6- a-p aranitrophenylthiopropionamido penicillanic acid, 6- a-parachlorophenylthiopropionamido penicillanic acid, 6- a-phenylthiobutyrarnido penicillanic acid, 6-( a-phenylthiocaproamido penicillanic acid, 6- a-phenylthioisovaleramido penicillanic acid, 6- a- (4-t-butylphenylthio propionamido] penicillanic acid, 6- wortho-tolylthiopropionamido penicillanic acid, 6- a-orth o-nitrophenylthiopropionamido) pe
  • Preparation 20 Preparati0n of potassium 6- [D -ecplzenoxybulyramido] thiopenicillanale Potassium 6-[D( -0t phenoxybutyramido]penicillanate (3.0 gm.) is dissolved in water (30 ml.) and layered with ethyl acetate. After cooling to 5 C.., dilute phosphoric acid (40%) is added until a pH of 2 is attained. The penicillin acid is extracted into the ethyl acetate and a further extraction with fresh ethyl acetate is made.
  • Skellysolve B is added to the solution with cooling and shaking until the solution becomes cloudy, and an oil separates.
  • the solution is decanted and flashed to dryness.
  • the residue is triturated with ether, and a white solid forms which is removed by filtration, washed with dry ether, dried in vacuo and found to weigh 2.5 gm.
  • the aqueous solution of 6-[D,L-a- Jhenoxypropionamido]thiopenicillanic acid is extracted 1nd re-extracted with trichlorethylene.
  • the combined :richl-orethylene extracts are dried over anhydrous magnesium sulfate in an ice bath and placed under vacuum for 30 minutes to remove hydrogen sulfide.
  • the magnesium sulfate is then filtered off and washed on the filter with trichlorethylene.
  • To the combined filtrates is added 70 ml.
  • Potassium 6- [D,L-ot-phenoxybutyramido] penicillanate (30.0 gm.) is dissolved in a mixture of 100 ml. ice Water and 100 ml. trichlorethylene. After cooling the mixture to 5 C., 6 N hydrochloric acid is added with stirring until a pH of 2 is attained. The trichlorethylene phase is separated and maintained at 5 C. The aqueous phase is extracted again with 50 ml. of trichlorethylene and the trichlorethylene extracts are combined, mixed with 40 gm. of anhydrous magnesium sulfate and stirred in an ice bath for 30 minutes.
  • the combined extracts are then filtered, and the filter cake is washed with ml. trichlorethylene.
  • the filtrates are placed together with 250 ml. dimethylformamide in a one-liter 3-necked flask with a drying tube vent and cooled to -3 C. in an ice-acetone bath.
  • To the chiiled filtrates is then added 10.2 ml. 2,6-lutidine and then ethylchloroformate, 8.7 ml. over a five-minute period after which the resulting solution of mixed anhydride is stirred in the ice bath for 30 minutes.
  • An anhydrous solution of 16.8 gm. of sodium hydrosulfide trihydrate in 150 ml.
  • dimethylformamide is mixed with the mixed anhydride solution over a period of five minutes and the resulting slurry is stirred for one hour at 0 C.
  • the slurry is then slowly decanted into 1500 ml. of ice water having a pH of 1.9.
  • the pH of the mixture. is maintained at about two by adjustment with 6 N; hydrochloric acid.
  • the aqueous solution of 6-[D,L-a-phenoxybutyramido] thiopenicillanic acid is extracted with trichlorethylene.
  • the combined trichlorethylene extracts are dried over anhydrous magnesium sulfate on an ice bath and placed under vacuum for 30 minutes to remove hydrogen sulfide.
  • the magnesium sulfate is then filtered 01f and washed on the filter with trichlorethylene.
  • To the combined filtrates is added ml. of a 22% solution of potassium ethylhexanoate in methyl isobutyl ketone whereupon the product, potassium 6-[D,L-ot-phenoxybutyramido]thiopenicil lanate, crystallizes out of solution.
  • the slurry is stirred for 20 minutes at room temperature and thereafter in an ice bath for minutes While the product crystallizes.
  • the product is collected by filtration, washed with trichlorethylene, vacuum-dried over P 0 for 18 hours, and thereafter the structure is confirmed by nuclear magnetic resonance and infrared absorption data.
  • Preparation 23 Preparati0n of 6-aminothiopenicillanic acid
  • a fermentation broth is prepared by the fermentation of Escherichia coli under submerged aerobic conditions according to conventional procedures and found to contain 5,270 penicillin amidase units per ml.
  • a solution of 2.5 gm. calcium nitrate dihydrate dissolved in 6 ml. of water is added to one liter of such fermentation broth. After mixing, the broth is filtered, and the filtered mat washed with 150 ml. water. The filtered mat is then suspended in 200 ml. of water to which is added 5 ml. of toluene, and the suspension is stirred for three hours. After stirring, the suspension is filtered and the collected solid materials washed with ml. of water. The filtrate is stirred with 1 gm. activated carbon (Darco KB) and 1.2 ml. of Quaternary Ammonium Salt Mixture No. I. Quaternary Ammonium Salt Mixture No.
  • I is commercially available from Armour & Company of Chicago, Ill., under the trademark of Arquad 16-50 and is a liquid quaternary ammonium salt mixture containing, by weight, about 45% hexadecyltrirnethylammonium chloride, about 3% octadecyltrimethylammonium chloride, about 2% octadecenyltrimethylammonium chloride, about 35% isopropanol, about 14% water, and about 1% sodium chloride.
  • Benzylthiopenicillin (8 gm.) is prepared by the proce-' dure described in United States Patent No. 2,751,378 and added to 400 ml. of penicillin amidase solution, prepared according to the procedure described above. This suspension is maintained at pH 8.0 and 35 C. for four hours, during which time the enzyme brings about the enzymatic hydrolysis of the benzylthiopenicillin.
  • the benzylthiopencillin and phenylacetate (produced during the hydrolysis) are removed at pH 2 and 5 C. with methyl isobutyl ketone. After extraction, a portion of the liquor (250 ml.) is adjusted to pH 7 with sodium hydroxide and vacuum concentrated to 20 ml.
  • the concentrate is adjusted to pH 4 with hydrochloric acid (6 N) and cooled to 5 C. and allowed to stand for 20 hours at 5 C. during which time crystallization occurs.
  • the crystals are filtered, washed with water (10 ml.) and then dry acetone.
  • the 6-aminothiopenicillanic acid is recovered, weighs 0.29 gm. and the presence of the o H -CSH group and ,B-lactam ring is confirmed by infrared analysis.
  • Preparation 24 Preparali0n 0 o-(a-aminophenylacetamido) -zhi0penicillanic acid 6 (a carbobenzyloxyaminophenylacetamido)penicillanic acid which is obtained by the reaction of equivalent quantities of 6- a-aminophenylacetamido penicillanic acid and benzyl chlorocarbonate in aqueous sodium hy-

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Description

United States Patent "Ce 3,316,273
Patented Apr. 25, 1967 3 316 273 Preferred compounds of the present invention are those of the formulae PENICILLlN ALDEHYDES William J. Gottstein and Lee c. Cheney, Fayetteville,
N.Y., assignors to Bristol-Myers Company, New York, 5 0 c c -c11 04311, H N.Y., a corporation of Delaware 5 l N0 Drawing. Filed Mar. 20, 1964, Ser. N0. 353,577
13 Claims. (Cl. 260306.7) This invention relates to novel antibacterial agents and f chemical reagents and, more particularly, to 6-substituted ArXC-C-NHOHOH C CH3 H penicillanyl aldehydes and especially to penicillin alde- LW L Q hydes.
The aldehydes of the present invention are effective antibacterial agents in vivo against Gram-positive bacteria 3 and are also effective agents for use in a search for oXida- 15 Z H tive enzymes.
There is thus provided, according to the present invention, the series of compounds having the formula wherein R is amino, acylamino, benzyloxycarbonylamino, ph-thalimido or tritylamino. In these compounds the Ar moiety may exist in tautomeric equilibrium with the enolic structure 0 I R -H--( JNH-(IJHC \CZCHB H -C=(|3OH N i RD O=CN-OHC=O H \O/ The preferred series of compounds is that of the O formula t /s\ /GH3 I 0 I s CH3 R2 Q NH JH 40 %NHCHO ofoH; 11 O=C -r N H7C=0 O=( )-N-CH-C 0 in which R represents the side chain of any of the known X s CH penicillins other than those containing a group which is H 3 altered by reaction with Raney nickel at 0 C. to 50 C. Ar-NHCNH(|3HCH ooH3 H as determined by simple test. It is this series which may be named 6- (substituted formamido)penicillanyl aldehydes or, more simply, penicillin aldehydes. S
In addition to the Well-knovvn, older penicillins pro- E Z H duced by the use of precursors in fermentation processes, other known penicillins include those disclosed in US. Patents 2,941,995, 2,951,839, 2,985,648, 2,996,601, 3,007,- 920, 3,025,290, 3,028,379, 3,035,047, 3,040,032, 3,040,033, 3,041,332, 3,041,333, 3,043,831, 3,053,831, 3,071,575, (C H CNHOH-CH 0-011 H 3,071,576, 3,079,305, 3,079,306, 3,080,356, 3,082,204, I, 3,093,547, 3,093,633, 3,117,119, 3,118,877, 3,120,512, 3,120,513, 3,120,514; in British patent specifications 6 874,414, 874,416, 876,516, 876,662, 877,120, 877,323, C 877,531, 878,233, 880,042, 880,400, 882,335, 888,110, S 1 888,552, 889,066, 889,069, 889,070, 889,168, 889,231, H 890,201, 891,174, 891,279, 891,586, 891,777, 891,938, I I 893,518, 894,247, 894,457, 894,460, 896,072, 899,199, 0 900,666, 902,703, 903,785, 904,576, 905,778, 906,383,
918,169, 920,176, 920,177, 920,300, 921,513, 922,278, 0 s CH3 924,037, 925,281, 931,567, 932,644, 938,066, 938,321, g \CiCH: H 939,708, 940,488, 943,608, 944,417; in numerous pub- 1 II I HAJZO lished Belgian patents, e.g. 593,222, 595,171, 597,859, 602,494, 603,703, 609,039, 616,419, 617,187; and South 1 0 S African patent applications, e.g. 60/2882, 60/3057, 60/ l C i H 3748, 61/1649, R61/2751, 62/54, 62/4920, 63/1612 and 63/2423. Za 0= NOHC=0 therein R represents hydrogen, amino, carbobenzoxymino, phenyl, fluoro, chloro, bromo, iodo, hydroxy, or lower)alkanoyloxy including especially acetoxy or (lowr)alkoxy; X represents oxygen or sulfur; R and R each epresent hydrogen, phenyl, benzyl, phenet-hyl or (lowr)alkyl; R represents (lower)alkyl; R and R each repesent (lower)alkyl, (lower)alkylthio, benzylthio, cyclo- 1 II Ar-C- and Ar represents the monovalent radical of the formula wherein R R and R are each a member selected from the group consisting of hydrogen, chloro, bromo, iodo, trifluoromethyl, phenyl, (lower)alkyl and (lower)alkoxy, but only one R group may represent phenyl.
The term (lower)alkyl as used herein means both straight and branched chain aliphatic hydrocarbon radicals having from one to ten carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl, 2-ethylhexyl, heptyl, decyl, etc. Similarly, where the term (lower) is used as part of the description of another group, e.g., (l0wer) alkoxy, it refers to the alkyl portion of such group which is therefore as described above in connection with (lower)alkyl.
Preferred embodiments of the present invention are the restricted series of compounds of the following formulae wherein R represents (lower) alkyl;
wherein R represents (lower)alkyl and R and R each represent a member selected from the group consisting of hydrogen and chloro;
wherein R is (lower)alkyl and R is a member selected I, from the group consisting of hydrogen and chloro; 0
o=o-N-- crr-o=o wherein R represents (lower)alkyl;
wherein R is (lower)alkyl; and the individual compounds of the following formulae O-G-C'H O=C-N--CI-IC=O The compounds of the present invention are prepared,- according to the present invention, by either of two methods, as follows:
METHOD A.-OXIDATION OF PENICILLIN ALCOHOLS The present penicillin aldehydes are prepared by oxidation of the corresponding penicillin alcohols by the addition of about 2-5 moles of a carbodiimide (e.g. dicyclohexylcarbodiimide, diisopropylcarbodiimide) to a solution of about one mole of the alcohol and 0.1-2.0 mole anhydrous phosphoric acid in dry dimethyl sulfoxide. Tetramethylene sulfoxide may also be used and the sulfoxide may be diluted with up to nine volumes of an inert solvent. The reaction proceeds readily at room temperature. The phosphoric acid functions as a catalyst and may be replaced with phosphorous acid, cyanoacetic acid or pyridinium phosphate, trifluoroacetate, hydrochloride or sulfate. Further details are given in the examples below.
METHOD B.--HYDROGENOLYYSIS BY RA'NEY NICKEL OF PENICILLIN THIOACIDS AND THIOESTERS The penicillin aldehydes of the present invention are preferably prepared by hydrogenolysis by Raney nickel of a thiopenicillin (also called a penicillin thioacid; e.g. see US. Patent 2,751,378) which is preferably in the form of the free acid or of a salt such as an alkali metal salt or an amine salt. The reaction is carried out at 50 C. and preferably at close to 0 C. It is advantageous to use an anhydrous or nearly anhydrous solvent such as tetrahydrofuran or acetone and to use catalytic amounts of a weak anhydrous acid such as glacial acetic acid as the penicillin aldehydes are very sensitive to bases. It is advantageous to add sufficient water to dissolve the thiopenicillin in the reaction mixture in the event that it is not completely soluble in the anhydrous solvent, The proportions are not critical but it is of course desirable to use enough Raney nickel to assure a high yield in the reaction; it is thus preferred to use a weight of Raney nickel (on a Wet basis) equal to at least three times the weight of the thiopenicillin. The activated Raney nickel available commercially is quite suitable for use as is that prepared according to Mozingo et al., J. Amer. Chem. Soc., 65, 1013 (1943). The thiopenicillins can be replaced, if desired, with their esters, e.g. with alkylthio or aralkylthio esters; see J. Amer. Chem. Soc., 75, 3636-3637 (1953), J. Chem. Soc. (London), 3733-3739 (1953) and Chem. Ber., 92, 530-534 (1959).
It is on occasion advantageous, but by no means essential, to conduct the hydrogenolysis in the presence of a trapping agent for the aldehyde being produced. By this is meant an N,N'-disubstituted-ethylenediamine or a 1,3-disubstituted-propane-1,3-diarnine. These reagents condense with the aldehyde formed during the course of the hydrogenolysis to form the corresponding imidazo lidines and hexahydropyrimidines respectively, thus protecting the aldehyde function from further reduction or other reaction, The penicillin aldehyde can be readily regenerated by treatment of these heterocyclic derivatives with an acid, e.g., with p-toluenesulfonic acid monohydrate in a suitable solvent such as acetone-ether.
Examples of preferred diamines for use as the aldehyde trapping agents are those of US. Patent 2,739,981 and especially N,N'-dibenzylethylenediamine [W. F. Minor, D. A. Johnson and L. C. Cheney, J. Org. Chem., 21, 528 (1956)], N,N-diphenylethylenediamine [W. Wanzlick and W. Lochel, Chem. Ber., 86, 1463 (1953)]. Other suitable diamines include N,N'-dimethy1ethylenediamine [A. J. Birch, J. Cymerman Craig and M. Slaytor, Australian J. Chem, 8, 512 (1955)] and N,N-bis- (p-methoxybenzyl)-1,3-diaminopropane [J. H. Billman and J. L. Meisenheimer, J. Med. Chem., 6, 682 (1963)].
The product is easily separated from the Raney nickel by filtration and from unreacted starting thiopenicillin by virtue of the fact that only the latter contains an acidic group and can thus be extracted from an organic solvent, such as ether, into aqueous alkali, e.g. 5% NaI-ICO Where desired, the penicillin aldehydes may be purified by reaction with 2,4-dinitrophenylhydrazine or N,N-di'benzylethylenediarnine [of J. Org. Chem, 21, 528-529 (1956) and US. Patents 2,717,893 and 2,767,168] to form the hydrazone or imidazolidine respectively, which can itself be purified as by recrystallization and then treated to regenerate purified penicillin aldehyde, e.g. by acid hydrolysis or aldehyde interchange.
The compounds of the present invention exhibit in vitro antibacterial activity. Thus when the Minimum Inhibitory Concentration (MIC) in meg/ml. was determined in heart infusion broth (to which 5% pooled human serum had been added) for the compounds having the structures Despite these low levels of in vitro antibacterial activity, t was surprisingly discovered that these compounds are :ffective antibacterial agents in vivo, that is, exhibited a ninirnum curative dose in 50% of the mice tested (CD against an overwhelmingly lethal dose of S. aureus Smith is follows:
Compound: CD in mgm./ kg. I 230 II 500 III 6.0 IV 3.4 Ampicillin 0.6
1 Complete protection (zero deaths) at 500 mgm./kg.
oxidative enzymes in the microorganism being examined.
The following examples are presented to illustrate the present invention but not to limit it. All temperatures are given in degrees centigrade.
EXAMPLE 1 O C H3 6-phenoxyacetamidopenicillanal. Commercial, pyrophoric Raney active nickel catalyst, 125 g. wet weight, was washed three times with 275 ml. portions of absolute ethanol and then with four 275 ml. portions of tetrahydrofuran (Tl-IF). The nickel was then suspended in 500 ml. of THF, 10.5 ml. (0.175 mole) of glacial acetic acid was added and the mixture was stirred and cooled to 4. A solution of 20.22 g. (0.0500 mole) of the potassium salt of penicillin V thiol acid was prepared by suspending the solid in 200 ml. THF and adding 15 ml. Water. This solution was added to the suspension of nickel and the mixture was stirred at 2 for 30 minutes. The catalyst was removed by filtration through diatomaceous earth (Supercel) which was then washed with five 150 ml. portions of THF. From the combined, reddish filtrates the solvent was removed in vacuo at 33. The residue was taken up in 1700 ml. ether which was then extracted with three 250 ml. portions of 5% NaHCO solution to remove unchanged starting material; the aqueous extracts were discarded. The ethereal phase was then washed with three 175 ml. portions of water, dried over sodium sulfate, decolorized with charcoal and filtered through Supercel. Complete removal by distillation of the solvent from the filtrate left the product, 6-phenoxyacetamidopenicillanal, as a stiff gum (5.7 g.). It was converted to a white solid by twice dissolving it in about 15 ml. methylene chloride which was then poured into 800 ml. n-pentane to precipitate 4.0 g. of the product as a white solid.
EXAMPLE 2 O-CHs The compounds of the present invention are useful agents for the detection of microorganisms containing oxidative enzymes, e.g. of the type used to oxidize steroids. Thus, cells of the microorganisms being investigated are grown for about 24 hours in a suitable medium (e.g. heart infusion broth with glucose, yeast-malt medium) on a shaker (e.g. at 28 or 37 C.). The cells from 10 ml. are obtained by centrifugation and added to 2 ml. of substrate solution at each of various pHs such as 5 and 7, e.g. to a solution of 500 mcg./ ml. phenoxymethylpenicillin aldehyde (IV) in 20% acetone-80% 0.2 molar pH 5 citrate bufier. That mixture of cells and substrate is then returned to the shaker for a short period of time (e.g. four hours). Aliquots are taken, diluted (e.g. ten-, fiftyand one hundred-fold) and assayed by a typical penicillin assay, e.g. by the disc method on an agar plate seeded with Penicillin V Aldehyde (G-Phenoxyacetamidopencillanal) (l) 6-phenoxacetamidopenicillanic thiol acid, N,N'-dibenzylethylenediamz'ne (DBED) salt.--A mixture of 38.0 g. (0.985 mole) of pencillin V potassium salt, 20.6 g. of triethylamine hydrochloride (0.15 mole) and 1400 ml. of methylene chloride was stirred for /2 hour at 25. Most of the methylene chloride was then removed under vacuum at 30. Dimethylformamide (DMF), 500 ml., was then added and the mixture was cooled to 4. Ethyl chloroformate, 9.38 ml. (0.0985 mole), was added and the temperature immediately rose to +2.5 The mixture was stirred and cooled down to 5 during the next ten minutes forming the mixed anhydride. A solution of 2 2 g. (0.2 mole) of sodium hydrosulfide (hydrated) in 350 ml. of DMF was prepared by stirring for ten minutes at room temperature under a nitrogen atmosphere. The solution was a turbid yellow, but all chunks of the salt had B. subtilis; activity in this assay indicates the presence of disappeared at this time. The solution of sodium hydro- .9 sulfide was poured into the mixed anhydride solution, and the resulting dark mixture was stirred at +l to 0 for ten minutes forming the thio acid. The reaction mixture was immediately poured into 3700 ml. of ice water, acidified to pH 2 with ca. 50 ml. of 6 N H 80 and extracted with two 1500 ml. portions of ice-cold ether. The combined yellow ether extracts were washed twice with 75 ml. portions of ice water, and then extracted with two 200 ml. portions of 4.25% NaHCO solution which carried the yellow color into the aqueous layer. The combined basic extracts were added to a solution of 36 g. (0.1 mole) of N,N'-dibenzylethylenediamine diacetate in 200 ml. of water forming the bis salt of the thio acid. The title compound crystallized when all of the dissolved ether was removed under vacuum. It was filtered off, dried briefly on the filter, then placed in a 2-liter round-bottom flask. Acetone, 100 mL, was added followed by 200 ml. of ethyl acetate and the mixture was flashed to dryness at 33 thereby removing the remaining traces of water. The yellow solid was partly dissolved in 50 ml. of dry acetone, diluted with one liter of dry ether and filtered off and dried under vacuum. Yield, 39.5 g. Its infrared spectrum showed NH (3350), intense B-lactam (1760), amide (1670 and 1495), -COS (1510) and phenyl absorptions at 1590, 750 and 695 cmr (2) 3-(1,3-dibenzyl 2-imitlazolidinyl)-2,2-dimethyl- 6-phenoxyacetamidopenam.Commercial Raney active nickel catalyst, No. 28, 25 g. (wet weight) was washed three times with 55 ml. portions of absolute ethanol, and then with four 55 ml. portions of tetrahydrofuran (THF). The washing procedure is speeded materailly if a bar magnet is used to pull the nickel to the bottom of the breaker after each wash. The nickel was then suspended in 100 ml. of THF and 2.1 ml. (0.035 mole) of glacial acetic acid and 12 g. (0.0050 mole) of N,N'-dibenzylethylenediamine (DBED) was added and the suspension was stirred and cooled to A cooled (5) solution of 4.86 g. (0.0100 mole) of pen V thiol DBED salt in 40 ml. of THF was added and the mixture was stirred at 2' for hour. The yield and quality of product may possibly be substantially improved by reducing this reaction time to about 15 minutes. The mixture was filtered over diatomaceous earth (Supercel) and the catalyst was washed with four 30 ml. portions of THF. The THF was removed under vacuum at 33 and the dark residue was dissolved in 300 ml. of ether. A considerable amount of gum failed to dissolve. On addition of 50 ml. of water, part of the gum dissolved. In some runs a pink solid separated at this point and was filtered off. The water layer was separated and the ether solution was extracted successively with two 30 ml. portions of 5% acetic acid, 50 ml. of water, three 30 ml. portions of 5% sodium bicarbonate and three 30 ml. portions of Water. The ether solution was dried briefly over sodium sulfate, treated with decolorizing carbon and filtered over Supercel which gave a nearly colorless filtrate. The solvent was flashed ofl at 33, the residue was flashed with two 100 ml. portions of chloroform to remove moisture and then thoroughly dried over phosphorus pentoxide at 0.5 mm. The yield of the titled compound was 2.23 g. of a hard gum.
(A similar procedure yielded the N,N-diphenylimidazolidinyl analog of the title compound which was obtained as a crystalline solid, M.P. 174176 sl. dec.)
Analysis.Calcd. for C H N O S: C, 68.15; H, 6.10. Found: C, 67.90; H, 6.20.
The infrared spectra of the two imidazolidines were very similar.
(3) 6-pherz0xyacetamidopenicillanuI.-A solution of 2.23 g. (4.00 millimole) of the above N,N'-dibenzylimidazolidine in 15 ml. of dry acetone and 45 ml. of ether was treated all at once with a solution of 1.67 g. (8.80 millimole) of p-toluenesulfonic acid monohydrate in 7.5 ml. of acetone and 22.5 ml. of ether. A heavy pre- 10 cipitate formed instantly. The suspension was stirred for 15 minutes at 25 and then filtered. Addition of 1 ml. of the same toluenesulfonic' acid solution to the filtrate produced no further precipitate, so the filtrate was extracted twice with 25 ml. portions of water (the first extract was strongly acidic, the second much less acidic), once with 20 ml. of 2% sodium bicarbonate and again with 20 ml. of water. The ethereal solution was dried over sodium sulfate, treated with decolorizing carbon, and filtered over Supercel. Removal of the solvent and through drying under vacuum yielded 1.10 g. of the product, 6-phenoxyacetamidopenicillanal, as a very hard, almost glassy gum. Its infrared spectrum (in methylene chloride) showed a weak OH (3500), NH (3370), intense B-lactam (1780), medium CHO (1730), intense amide (1690 and 1510), phenyl (1595, 1590 and 1490) and phenyl-OC (1230). Its NMR spectrum (in CDCl showed all of the expected sharp resonance lines.
EXAMPLE 3 s CH3 6-(3-phenyl-5-meflzylfsoxazole 4 carboxamido)penicillamal.-One hundred grams of wet No. 28 commercial Raney Ni (Raney Co.) was washed three times with ml. portions of absolute ethanol, three times with 100 ml. portions of acetone and finally a fourth portion of 100 ml. of acetone was added and the slurry cooled and stirred at 0 to 5 C. while 7 g. of N,N-diphenylethylenediamine, 2.4 ml. of glacial acetic acid and a solution of 10 g. of freshly prepared triethylammonium thiol oxacillin [triethylammonium 6-(5-methyl-3-phenylisoxazole-4-carboxamido)thiopenicillanate] in 50 ml. of acetone was added in that order. These were added at such a rate as not to exceed 5 C. The slurry was stirred /2 hour at 2 C., filtered and the Raney Ni sludge washed with 4 50 ml. portions of acetone. The combined filtrates were evaporated to dryness at 25 C. under reduced pressure and the resulting oil dissolved in 500 ml. of dry ether, filtered and the ether washed with 3 X 100 ml. of 5% H PO 3X 100 ml. H O, 3x 100 ml. of 5% NaHCO and 3 X 100 ml. water. The ether solution was then dried briefly over Na SO filtered and treated with a concentrated solution of p-toluenesulfonic acid in acetone until the solution was strongly acid to moist pH paper after stirring for 5 minutes. The crystalline amine salt was then filtered off and the ether filtrate washed with 3 X 100 ml. water, 3 100 ml. 5% NaHCO and 3X 100 ml. of water. The ether solution was dried over Na SO briefly; filtered and strippel to dryness under high vacuum until the product, 6-(3-phenyl-S-methylisoxazole-4-carboxamido)penicillanal, was obtained as a frothy solid vlgich was scraped out of the flask and found to weigh Analysis.-Calcd. for C I-I N O S: C, 59.25%; H 4.96%; N, 10.9%.
Found: C, 59.70%; H, 5.61%;
Penicillin G aldehyde (6 phenylacetamidopeniciL lanal).-To a precooled, stirred suspension of 13.08 g. (0.03 mole) of penicillin G-triethylammonium salt in 200 ml. of dimethylformamide (DMF) was added 3.24 g. (0.03 mole) of ethyl chloroformate dropwise over a ZOminute period. The resulting nearly clear solution was then treated all at once with 3 g. of NaSH in 50 ml. of DMF which had been previously dissolved by pulverizing in a mortar and stirred and cooled in an ice bath until a cloudy solution had been obtained. The resulting dark green reaction mixture was stirred one-half hour with the ice bath removed and poured into a mixture of 500 ml. benzene, one liter of crushed ice in water and 30 ml. of 40% H PO The benzene extract was washed three times with cold water and partially dried by filtering through sodium sulfate. The benzene was then removed under reduced pressure on the flash evaporator and when the benzene was nearly all removed (avoiding heating) the resulting oil was dissolved in 50 ml. of dry acetone and added immediately, dropwise, to a previously prepared stirred suspension of 100 g. of No. 28 commercial Raney Ni (Raney Co.), [previously washed 3X with absolute ethanol and 4X with dry acetont (200 ml. portions)], 300 ml. of acetone, 10.5 g. of N,N'-di-phenylethylenediamine and 3.6 ml. of glacial acetic acid at 2.0 C. The addition took five minutes and the temperature rose to 4 C. After stirring an additional A hour the Raney Ni was filtered off through Supercel filter aid and the Raney Ni sludge was washed 4 with 100 ml. portions of acetone. The combined filtrates were evaporated to an oil on the flash evaporator and dissolved in one liter of ether; a precipitate was then filtered off and the ether solution was extracted 3 with 5% H PO (100 ml.) 3X with Water (100 ml.), 3X with 5% NaHCO (100 ml.) and 3 X with water. The ether solution was dried briefly over Na SO filtered and evaporated under reduced pressure to an oil. The oil was then dissolved in 100 ml. of acetone and 900 ml. of ether added. To this solution was added 6 g. of p-toluenesulfonic acid trihydrate dissolved in 50 ml. of acetone and the resulting slurry stirred for ten minutes, filtered and the filtrate washed three times with 200 ml. of water. The ether solution was dried briefly over Na SO filtered, stripped to dryness and dissolved in 300 ml. of CH Cl and stripped again followed by pumping at high vacuum (0.1 mm. Hg). The resulting product, penicillin G aldehyde, was thus obtained as a frothy solid which was scraped out, dried over P under vacuum and found to weight 5.6 g.
Alternate procedure.Same as above except the potassium salt of penicillin G-thiol acid was used; yield, 1 g.
Analysis.-Calcd. for C H N O S: C, 60.3%; H, 5.70%; N, 8.79%. Found: C, 59.8%; H, 6.18%; N, 8.03%.
Alternate p r0cedure.-Same as above except N,N'-dibenzylethylenediamine was used in place of N,N-diphenylethylenediamine and dilute (2%) acetic acid was used in place of H PO in the acid washes. Tetrahydrofuran was used as solvent for the reaction and the DBED salt of the thiol-penicillin G was used. The yield was 27 g. (60%) of the crude dibenzylimidazolidine and the aldehyde was liberated as before with p-toluenesulfonic acid. The aldehyde was the same as in previous runs by infrared spectrum. The yield in this experiment was approximately 50%, an improvement over the methods given above.
EXAMPLE 5 R s CH3 0C 1H-o-NHCHC C CHa H CH. 0=( 3-I I-( 1Ho o Phenethicillin aldehyde (6 or. phenoxypropionamidopenicillanal or or-phen0bcyethylpenicillin aldehyde).-Fifty grams of wet Raney nickel was washed three times with 100 ml. portions of absolute ethanol and three times with 100 ml. portions of dry acetone and finally 100 ml. of dry acetone was added and, while cooling to 0 there was added 4.24 g. (0.02 mole) N,N'-diphenylethylenediamine and 2.4 ml. glacial acetic acid. To this cooled and stirred slurry there was added all at once a cooled partial solution of 9.4 g. (0.02 mole of potassium thio-phenethicillin (potassium 6-ot-phenoxypropionamido-thiopenicillanate) in 100 ml. acetone and 2.4 ml. glacial acetic acid. The temperature rose to 6 C. and after stirring 30 minutes at 0 C. the Raney nickel was removed by filtration and washed four times with 50 ml. portions of acetone. The combined filtrates were flash-distilled to dryness under reduced pressure at 20 C. to leave the expected imidazole as a purple oil which was triturated with 500 ml. dry ether and filtered. The ethereal filtrate was washed three times with ml. portions of water, three times with 100 ml. portions of 2% NaHCO solution and three times with water and dried over Na SO To the dry ethereal filtrate there was then added a solution of p-toluene sulfonic acid (5 g. in 50 ml. acetone) until the solution was acidic to wet pH paper. The solid p-toluenesulfonic acid salt of N,N-diphenylethylenediamine which precipitated was removed by filtration and the filtrate was treated as before with the acid repeatedly until no more solid formed upon addition of acid and the ethereal solution was still acidic to wet filter paper after standing ten minutes. The ethereal solution containing the product, phenethicillin aldehyde, was then washed with 2% NaHCO and water, dried, filtered and the solvent was removed by distillation to dryness under high vacuum to leave the product as a frothy, amorphous solid weighing 2.9 g. and having an infrared absorption spectrum consistent with the structure given above. I
Analysis.-Calcd. for C H N O S: C, 58.6; H, 5.80; N, 8.06. Found: C, 58.9; H, 6.13; N, 7.52.
Benzyloxypenicillin aldehyde (6 N carbobenzyloxyaminopenicillanaL-To 45 g. of previously acetonewashed Raney nickel suspended in 100 ml. of acetone with 3.7 g. (0.0174 mole) N,N'-diphenylethylenediamine and 2.1 m1. of glacial acetic acid, was added 7 g. (0.0174 mole) of benzyloxypenicillin thiol (potassium salt) dissolved in 25 ml. of acetone and 3 ml. of glacial acetic acid. The mixture was stirred for one hour in an icebath and the mixture was filtered. The filtrate was evaporated to an oil under reduced pressure at 30 and residue was sl-urried with 100 ml. of ether. The ether was washed twice with dilute sodium bicarbonate and several times with water. The ether was dried over anhydrous magnesium sulfate. The drying agent was separated and the ether solution treated with a solution of p-toluenesulfonic acid in acetone until there was no more turbidity. The N,N'-diphenylethylenediamine toluenesulfonate was collected and the filtrate was washed with water and dried over anhydrous MgSO The solvent was evaporated under reduced pressure to yield 1.5 g. of neutral benzyloxypenicillin aldehyde.
EXAMPLE 7 6 phthalimidopenicfllin aldehyde (6 phthalimidopenicillanal).T0 a solution of 5.0 g. (0.015 mole) of 6-phthalimidopenicillanyl alcohol in 50 ml. of anhydrous dimethylsulfoxide was added 10.0 g. (0.049 mole) of dicyclohexylcarbodiimide and 1.5 g. (0.0075 mole) of pyridinium trifiuoracetate. The mixture was swirled in an Erlenmeyer flask for a few seconds, placed in a desiccator under a nitrogen atmosphere, allowed to stand at 25 C. for 17 hours and then filtered and the filtrate thrown into 500 ml. of 50:50 chloroform-water and shaken. The chloroform layer was separated, washed with water and dried over MgSO for two hours. The chloroform solution was then filtered and evaporated to EXAMPLE 8 6-(5 methyl 3 0-chloropherzylis0xaz0le-4-carb0xamido)penicillanal [S-methyl-3-0-chl0r0phenyl 4 isoxazolylpenicillin rzIzlehyde.Twenty-five g. (wet weight) of Raney nickel, No. 28 was washed three times with 55 ml. portions of 100% ethanol, followed by four washings with 55 ml. portions of tetrahydrofuran (THF). The nickel was then suspended in 100 ml. THF and cooled to 4 and to this there was added 2.1 ml. glacial acetic acid. Potassium 6-(5-methyl-3-o-chlorophenylisoxazole-4-carboxamido)thiolpenicillanate (4.5 g.) was dissolved completely in 40 m1. THF and added to the Raney nickel. The reaction mixture was stirred below 0 for 15 minutes. The Raney nickel was then filtered through diatomaceous earth and washed five times with 60 ml.
portions of THF. The filtrate was evaporated to dryness.
The resulting oil was partially dissolved in 350 ml. ether. A pink gum appeared which was carried along with the ether. The ether was then extracted three times with 60 m1. portions of 5% NaHCO and three times with 40 ml. portions of water. The ether layer was then separated from the gum and dried over magnesium sulfate and charcoal. After filtration the ether was evaporated to dryness. The resulting oil could be solidified by dissolving it in methylene chloride or ether and precipitating with n-pentane. The product, 6-(5-methyl-3-o-chlorophenylisoxazole-4-carboxamido)penicillanal [5 methyl- 3-o-chlorophenyl-4-isoxazolylpenicillin aldehyde], precipitated as an amorphous solid, yield 150 mgm. Both IR and NMR spectra showed all of the expected absorptions.
EXAMPLE 9 In the procedure of Example 5 the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids 6-[a-(2-ch1orophenoxy)propionamido] thiopenicillanic acid, 6-[a-(4sultamylphenoxy)-n-butyramido] thiopenicillanic acid,
6- [a-(3,4-dimethoxyphenoxy) -n-pentanoamido] thiopenicillanic acid,
6-[11-(3 methylphenoxy isovaleramido] thiopenicillanic acid,
6-[ot-( l-methylthiophenoxy)propionamido] thiopenicillanic acid,
6-[u-4dirnethylaminophenoxy)-nhexanoamido] thiopenicillanic acid,
6-[a-(2-methoxyphenoxy)-ndecanoarnido] thiopenicillanic acid,
6- rz- 2,4dichlorophenoxy phenylacetamido] thiopenicillanic acid,
6-[a-(2-nitrophenoxy)-fi-phenylpropionamido] thiopenicillanic acid,
6-[a-(2-acetamidophenoxy)-'y-phenylbutyramido] thiopenicillanic acid,
6 OL- 2,4-dimethylphenoxy) -n-butyramido] thiopenicillanic acid,
6 [0L- 4-isopropylphenoxy propionamido] thiopenicillanic acid,
6- a- 3 -bromophenoxy) -n-butyramido] thiopenicillanic acid,
6-[u- (2-iodophenoxy)phenylacetamido] thiopenicillanic acid,
6-[a-(Z-diethylaminophenoxy)isovaleramido] thiopenicillanic acid,
6[w(3,5-dichlorophenoxy)isohexanoamid-o] thiopenicillanic acid,
6- oz- 4-cyclohexylphenoxy propionamido] thiopenicillanic acid,
6-[ot-phenoxy-isovaleramido] thiopenicillanic acid,
6-[aphenoxy-n-decanoamido] thiopenicillanic acid,
6 [or-phenoxy-v-phenylbutyramido] thiopenicillanic acid,
6- [a- 2-benzylphenoxy) -n-butyramido] thiopenicillanic acid,
6[a-(Z-trifluorornethylphenoxy)propionamido] thiopenicillanic acid, and
6- ot-(4-fluorophenoxy) propionamido] thiopenicillanic acid,
respectively, to produce 6- Ot- 2-chlorophenoxy propionamido] penicillanal,
6- cc- (4-sulfamylphenoxy) -n-butyramid-o penicillanal,
6-[a-(3,4-dirnethoxyphenoxy) -n-pentanoamido] penicillanal,
6- w 3 -methylphenoxy) isovaleramido] penicillanal,
6- [w (4-methylthiophenoxy propionamido] penicillanal,
6- oc- 4-dimethylaminophenoxy) -n-hexano amido] penicillanal,
6- [a- Z-methoxyphenoxy) -n-decanoamido] penicillanal,
6-[a-(2,4-dichlorophenoxy)phenylacetamido] penicillanal,
6- u- Z-nitrophenoxy) -,B-phenylpropionamido] penicillanal,
6- OL- 2-acetamidophenoxy) -'y-phenylbutyramido] penicillanal,
6- 0L- 2,4-dimethylphenoxy) -n-butyran1i-do] penicillanal,
6- a-(4-isopropylphenoxy)propionamido] penicillanal,
6-[a-(3-bromophenoxy)-n-butyramido] penicillanal,
6- a- 2-iodophenoxy phenylacetamido] penicillanal,
6-[a-(Z-diethylaminophenoxy)isovaleramido] penicillanal,
6- [oc- 3,5 -dichlorophenoxy isohexano amido] penicillanal,
6- 0!.- (4-cyclohexylphenoxy) propionamido] penicillanal,
6-[u-phenoxy-isovaleramido] penicillanal,
6-[m-phenoxy-n-decanoamido] penicillanal,
6- [or-phenoxy-'y-phenylbutyramido] penicillanal,
6- a- Z-b enzylphenoxy) -n-hutyramido] penicillanal,
6- OL- Z-trifluoromethylphenoxy) propion amido] penicillanal, and
6-[a-(4-fiuorophenoxy)propionamido] penicillanal,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system.
1 EXAMPLE In the procedure of Example 5 the potassium thio- Jhenethicillin is replaced by an equimolar amount of the aotassium salt of the acids,
5- ot-phenylthiopropionamido thiopenicill anic acid,
5- a-paranitrophenylthiopropionamide thiopenicillanic acid,
5- u-parachlorophenylthiopropionamido thiopenicillanic acid,
5- a-phenylthiobutyramido thiopenicillanic acid,
6- a-phenylthiocapro amide thiopenicillanic acid,
6- a-phenylthioisovaleramido thiopenicillanic acid,
6- a- 4-t-butylphenylthio propionamido] thiopenicillanic acid,
6-[a-ortho-tolylthiopropionamido]thiopenicillanic acid,
6 a-ortho-nitrophenylthiopropionamido thiopenicillanic acid,
6- u-parachlorophenylthiobutyramido thiopenicillanic acid,
6- oc- 3 ,4,5-trichlorophenylthio propionamide] thiopenicillanic acid,
6- a- 3 -trifluoromethylphenylthio butyramido] thiopenicillanic acid,
6- a-p arabromophenylthioisovaleramido) thiopenicillanic acid,
6- a-paraphenylphenylthiopropionamido) thiopenicillanic acid,
6- a- 4-methoxypheny1thio) caproamido 1 thiopenicillanic acid,
6- OL- 4-cyclohexylphenylthio butyramido] thiopenicillanic acid,
6- a-phenylthio-a-cyclohexylacetamido thiopenicillanic acid,
6- a-phenylthio-a-cyclopentylacetamide thiopenicillanic acid,
6- a- 2,4-dichlorophenylthio capro amide] thiopenicillanic acid,
6- oc- 2,4-diisoamylphenylthio propionamido] thiopenicillanic acid 6- w 4-benzy1phenylthio propionamido 1 thiopenicillanic acid,
6- [u-(4-su1famy1phenylthio )butyramido] thiopenicillanic acid,
6- [ot-(2-al1yloxyphenylthi0 )propionamido] thiopenicillanic acid,
6- oc- 4-ally1phenylthio isovaleramido 1 thiopenicillanic acid,
6- u- (4-dimethylaminophenylthio propionamido] thiopenicillanic acid,
6- a- 2,5 -dichloropheny1thio butyramido] thiopenicillanic acid,
6- [a- 2-iodophenylthio propionamido 1 thiopenicillanic acid,
6- oc- 2-acetamidophenylthio propionamido] thiopenicillanic acid,
6- oc- 4-diethylaminophenylthio propionamido] thiopenicillanic acid, and
6- a- 3-fiuorophenylthio butyramido] thiopenicillanic acid,
respectively, to produce 6-( a-phenylthiopropionamido penicillanal,
'6- a-paranitrophenylthiop ropionarnido pencillanal,
6- ot-parachlorophenylthiopropionamido) penicillanal,
6- oc-PhGllYl'thiOblltYlflIIlidO penicillanal,
6- a-phenylthiocaproamido penicillanal 6- a-phenylthioisovaleramido penicillanal,
6- u- 4-t-butylphenylthio propionamido 1 penicillanal,
6- u-ortho-tolylthiopropionamido 1 penicillanal,
6- a-ortho-nitrophenylthiopropionamido penicillan a1,
6- a-parach10rophenylthiobutyramido penicillanal,
6- oc- 3 ,4,5 -trich1orophenylthio propionamido] penicillanal,
6- oc- 3 -trifiuoromethylphenylthio butyramido] penicillanal,
6- u-parabromophenylthioisovaleramido penicillanal,
6- a-paraphenylphenylthiopropionamido penicillanal,
6- oc- 4-methoxyphenylthio caproarnido 1 penicillanal,
6- a-phenylthio-u-cyclohexylacetamido penicillanal,
6- a-phenylthio-u-cyclopentylacetamido penicill anal,
6- zx- (2,4-dichloropheny1thio caproamido1penicillanal,
6- ot-(2,4-disoamylphenylthio propionamido] penicillanal,
6- a-(4-benzylphenylthio propionamido1penicillanal,
6- a- 4-sulfamylphenylthio butyramido 1 penicillanal,
6- [ot-(2-ally1oxyphenylthio)propionamido]penicillanal,
6- [a- (4-allylpheny1thio isovaleramido1penicillanal,
6- a- 4-dimethylaminophenylthio propionamido] penicillanal,
6- [oc- 2,5 -dichlorophenylthio butyramido 1 penicillanal,
6- oc- 2-iodophenylthio propionamido penicillanal,
6- a- Z-acetamidophenylthio propionamido 1 penicillanal,
6- [a-( 4-diethylaminophenylthio) propionamido] penicillanal, and
6- [ca-( 3-fluorophenylthio butyramido1penicillana1,
respectively, each of which is is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system.
EXAMPLE 1 1 In the procedure of Example 5 the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
D,L-6- [ot-amino- 3 -thienyl acetamido] thiopenicillanic acid,
6- a-amino- 5 -ethyl-2-thieny1 acetamido 1 thiopenicillanic acid,
6- [0L-aI'1'lil'10-( S-methyl-Z-thienyl) acetamido] thiopenicillanic acid,
6- a-amino- 5 -t-butyl-2-thienyl) acetamido] thiopenicillanic acid,
6- [a-amino- 2,5 -dimethyl-3-thienyl) acetamido] thiopenicillanic acid,
6-[a-amino- (5 -chloro-2-thienyl) acetamido] thiopenicillanic acid,
6- a-amino- (5 -bromo-2-thienyl) acetamido] thiopenicillanic acid,
6- a-amino- 5 -phenyl-3 -chloro-2-thienyl) acetamido] thiopenicillanic acid,
6- [cc-amino 3 ,5 -dimethyl-2-thienyl acetamido] thiopenicilianic acid 6- a-amino 5 -cyclohexyl-2-tl1ienyl acetamido] thiopenicillanic acid,
6- ot-amino- 5 -diethylamino2-thienyl) acetamido] thiopenicillanic acid,
6- a-amino- 4-methylsulfony1-2-thienyl) acetamido] thiopenicillanic acid,
6- [a-amino- 3 -ethylthi0-2-thienyl acetamido] thiopenicillanic acid, and
6 [a amino (4 cycloheptyloxy 2 thienyl)acetamido]thiopenicillanic acid,
respectively, to produce 23 (N,N-tetramethylene-N'-phthalamido penicillanal, {N-(Z-phenylethyl) -N'-phthalarnido1penicillanal, (N,N-pentamethylene-N'-phthalamido penicillanal, -(N-isopropyl-N-phthalamido penicillanal, (N-u-methylbenzyl-N-phthalamido penicillanal, -(N-hexamethylene-N'-phthala'mido) penicillanal, -(N-tet rahydrofurfuryl-N'-phthalamido penicillanal, -(N-morpholino-N'-phthalamido penicillanal,
- [N (2,6 dimethylmor pholino) -N'-phthalamido penici'llanal, (N,N-di-n-butylphthalamido penicillanal, [o (2-methyl-1,2,3 ,4-tetrahydroquinolyl-carb onyl) benzamido1penicillanal, (N,N-diallyl-N-phtha1amid0 penicillanal, (N,N-diisopropyl-N-phthalamido penicillanal, (N,N-diethyl-N-phthalamido penicillanal, (N,N-dimethyl-N'-phthalamido penicillanal, (N-dodecyl-N-phthalamido penicillanal, (N-t-butyl-N'qahthalamido penicillanal, -N-methyl-N-phthalamido penicillanal, (N-ethyl-N'-phthalamido penicillanal, N-isobutylphthalamido penicillanal, (N-a-methylpropylphthalamido penicillanal, N-a-methylphenethyLN'-phthalaimido penicillanal, N-2-methoxyphenyl-N'-phthaiamido penicillanal, (N-phenyl-N-ethyl-N-phtha1a1nido penicillanal, [N- Z-methyl-S -ethy1piperidino) -N --phthalarnido] penicillanal, N-pyrrolidino-N-phth alamido penicillanal, -[N-(2-methylpyrrolidino)-N-phthalamido1penicillanal, [N- 2,5 -dimethylpyrr-olidino) -N'-phthalamido1 penicillanal, (N,N-heXamethylene-N'-phthalamido penicillanal, i-[N-( 1,1,3 ,3-tetrarnethylbutyl) -N-phthalamido] penicillanal, i- (N-cyclohexyl-N'-phthalamido penicillanal, i- [N- (3 -morpholinopropyl) -N-phthalamido1penicillanal,
and i- [N-(Z-methylpiperidino -N'-phthalamido1penicillanal,
espectively, each of which is isolated as a solid which s converted to an active antibacterial agent upon exposure an oxidative enzyme system.
EXAMPLE 16 In the procedure of Example 5, the potassium thioahenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
- oc- 2-chlorophenoxy) acetamido 1 thiopenicillanic acid,
3-[ot-(4-SU1-f2lII1Y1Ph6IIOXY) acetamido] thiopenicillanic acid,
5- a- 3 ,4-dimethoxyphenoxy) acetamido 1 thiopenicillanic acid 5- a- 3 -methylphenoxy) acetamido 1 thiopenicillanic acid,
5- oc- (4-rnethylthiophenoxy) a-cetamido thiopenicillanic acid,
5- u-4-dimethylaminophenoxy) acetamido 1 thiopenicillanic acid.
5- a-(2-methoxyphenoxy) acetamido] thiopenicillanic acid,
5- [a- 2,4-dichlorophenoxy) phenoxyacetamido] thiopenicillanic acid,
6- oc- 2-nitrophenoxy) acetamido] thiopenicillanic acid,
6- a- Z-acetamidophenoxy) acetamido] thiopenicillanic acid,
6- u- (2,4-dimethylphenoxy) acetamido] thiopenioillanic acid,
6- a- (4-isopropylphenoxy) acetamido thiopenicillanic acid,
6- oc- 3-bromophenoxy) acetamido1thiopenicillanic acid,
6- oc- (2-iodophenoxy phenoxyacetamido 1 thiopeniciilanic acid,
6- oc- Z-diethylaminophenoxy) acetamido 1 thiopenicillanic acid,
6- a- 3 ,5 -dichlorophenoxy) acetamido] thiopenicillanic acid,
6- 0C- 4-cyclohexylphenoxy) acetamido 1 thiopenicillanic acid,
6- a-phenoxyacetamido thiopenicillanic acid,
6- [a-(Z-benzylphenoXy) acetamido] thiopenicillanic acid,
6- oz- (Z-trifiuoromethylphenoxy) acetamido 1 thiopenicillanic acid,
6- [a-(4chlorophenoxy) acetamido] thiopenicillanic acid,
6-[a-(4-nitrophenoxy) aceta mido1thiopenicillanic acid,
6- 1 a- (4-bromophenoxy) acetamido]thiopenicillanic acid,
6- [a-(4-t-butylphenoxy) acetamido] thiopenicillanic acid,
6- [oc- (4-trifiuoromethylphenoxy) acetamido1thiopenicillanic acid,
6-[ot-(3-fluorophenoxy) acetamido]thiopenicillanic acid,
6- u- (4-sulfamylphenoxy) acetamido] thiopenioillanic acid,
6- [oc- 2-benzylphenoxy) acetamido 1 thiopeni'cillanic acid,
6- oc- 3 -methoxyphenoxy) acetamido] thiopenicillanic acid,
6-[a-(2-iodophenoxy)acetamido]thiopenicillanic acid,
6- oc- (3 -diethylaminophenoxy) acetamido 1 thiopenicillanic acid, and
6- oc- (2,4-diisoamylphenoxy) acetamido] thiopenicillanic acid,
respectively, to produce 6- a- 2-chlorophenoxy) acetamido1penicillanal,
6- [a-(4-sulfarnylphenoxy) acetamido1penicillan'al,
6- [oz-( 3 ,4-dimethoxyphenoxy) acetamido1penicillanal,
6- a- 3 -methylphenoxy) acetamido1penicillanal,
6[a-(4-methylthiophenoxy) acetamido1penicillanal,
6- oc- (4-dimethylaminophenoxy) acetamido 1 penicillanal,
6- [w (Z-methoxyphenoxy) acetamido1penicillanal,
6- a- 2,4-dichlorophenoxy) phenoxyacetarnido] penicillanal,
6- a- 2-nitrophenoxy) acetamido] penicillanal,
6- :x- 2-acetamidophenoxy) aceta mido1penicillanal,
6- cc- 2,4-dirnethylphenoxy acetamido 1 penicillanal,
6- [oc- (4-isopropylphenoxy) acetamido1penicillanal,
6- OC- 3 -bromophenoxy) acetamido1penicillanal,
6- a- 2-iodophenoxy) phenoxyacetamido] penicillanal,
6- [a- (Z-diethylaminophenoxy) acetamido1penicillanal,
6- a- 3,5 -dichlorophenoxy) acetamido 1 penicillanal,
6- oc- 4-cyclohexylphenoxy acetamido1penicillanal,
6- a-phenoxyacetamido penicillanal,
6- oc- 2-benzylphen0xy acetamido] penicillanal,
6- ov- (Z-trifluoromethylphenoxy acetamido] penicillanal,
6- oc- (4-chlorophenoxy) acetamido] penicill anal,
6- [a- (4-nitrophenoxy) acetamid01penicillanal,
6- [ot-(4-bromophenoxy acetamido1penicillanal,
6- [a-(4-t-butylphenoxy)acetamido1penicillanal,
6- [a-(4-trifiuor0methylphenoxy) acetamido1penicillanal,
6- oc- 3 -fluorophenoxy) acetamido1penicillanal,
6- [oc- (4-sulfamylphenoxy) acetamid01penicillanal,
6- [or-(l-benzylphenoxy) acetamido1penicillanal,
6- [a-(3-methoxyphenoxy) acetamido1penicillanal,
6- [a- (2-i0dophenoxy) acetamido1penicillanal,
6- a- 3 -diethylaminophenoxy) acetamido1penicillanal,
and
6- [a-(2,4-diisoamylphenoxy) acetamido1penicillanal,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxi-dative enzyme system.
EXAMPLE 17 In the procedure of Example 5, the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
6- a-phenylthioacetamido thiopenicillanic acid,
6- oc-p aranitrophenylthioacetamido thiopenicillanic acid,
6- a-parachlorophenylthioacetamido )thiopenicillanic acid,
6- oc- (4-tertiary-butylphenylthio) acetamido] thiopenicillanic acid,
6- a-ortho-tolylthioacetamido thiopenicillanic acid,
6- a-ortho-nitrophenylthioacetamido) thiopenicillanic acid,
6- a-( 3,4,5 -trichlorophenylthio acetamido] thiopenicillanic acid,
6- a- (3 -trifiuoromethylphenylthio acetamido] thiopenicillanic acid,
6- a-parabromophenylthioacetamido thiopenicillanic acid,
6- a-paraphenylphenylthioacetamido thiopenicillanic acid,
6- [a- (4-methoxyphenylthio acetamido]thiopenicillanic acid,
6- a- (4-cyclohexylpheny1thio acetamido] thiopenicillanic acid,
6- [a-2,4-dichlorophenylthio acetamido thiopenicillanic acid,
6- [oe-(2,4-diisamylphenylthi0) acetamido1thiopenicillanic acid,
6- [a- 4-benzylphenylthio) acetamido] thiopenicillanic acid,
6- [a-(4-sulfamylphenylthio) acetamido] thiopenicillanic acid,
6- [a- (2allyloxyphenylthio) acetamido] thiopenicillanic acid,
6'[a(4-a1lylphenylthio acetamido]thiopenicillanic acid,
6- [u- (4-dimethylaminophenylthio) acetamido1thiopenicillanic acid,
6- [ix- 2,5 -dichloropheny1thio acetamido] thiopenicillanic acid,
6- [a-(Z-iodophenylthio) acetamido1thiopenicillanic acid,
6- [a- (Z-acetamidophenylthio) acetamido] thiopenicillanic acid,
6- a- (4-diethylaminophenylthio acetamido] thiopenicillanic acid, and
6- [cz- (3-fluorophenylthio) acetamido] thiopenicillanic acid,
respectively, to produce 6- u-phenylthioacetamido penicillanal,
6- a-paranitrophenylthioacetamido penicillanal,
6-( a-parachl0rophenylthioacetarnido)penicillan a1,
6- [oc- (4-tertiary-butylphenylthio) acetamido1penicillanal,
6- a-ortho-tolylthioacetamido penicillanal,
6- wortho-nitrophenylthioacetamido) penicillanal,
6-[11- (3 ,4,5 -trichlorophenylthio) acetarnido] penicill anal,
6- [a-(3-trifluoromethylphenylthio) acetamido1penicillanal,
6-( a-parabromophenylthioacetamido penicillanal,
6- a-paraphenylphenylthioacetamido)penicillanal,
6- [oc- (4-methoxyphenylthio) acetamido1penicillanal,
6- [a- (4-cyclohexylpheny1thio) acetamido1penicillanal,
6- [a- 2,4-dichloropheuylthio acetamido] penicillanal,
6- [oc- (2,4-diisoamy1phenylthio acetamido penicillanal,
6- [a- (4-1bSHZY1PhEI1Y1thiO) acetamidoJpenicillanal,
6- [oc- 4-sulfarnylphenylthio) acetamido] penicillanal,
6- a- 2-allyloxyphenylthio) acetamido] penicillanal,
6-[a-(4-al1ylphenylthio) acetamido1penicillanal,
6- oc- 4-dimethylaminophenylthio acetarnido] penicillanal,
6- [oc- 2,5 -dichlorophenylthio acetamido] penicillanal,
6- [oc- (2-iodophenylthio) acetamido1penicillanal,
6- [0t- Z-acetamidophenylthio acetamido] penicillanal,
6- [a- (4-diethylaminophenylthio) acetamido1penicillanal,
and
6- [w 3-fluorophenylthio acetamido] penicillanal,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon eX- posure to an oxidative enzyme system.
EXAMPLE 18 6-[a-(4-nitrophenyl) acetamido]thiopenicillanic acid, 6- cc- (4-bromopheny1 acetamido] thiopenicillanic acid, 6- [u-(4-t-butylphenyl) acetamido1thiopenicillanic acid,
26 6- [a- (4-trifiuoromethylphenyl) acetamido1thiopenicillanic acid, 6- [w 3-fluorophenyl) acetamido] thiopenicillanic acid, 6-[u-(4-sulfamylphenyl) acetamido]thiopenicillanic acid, 6- [a-( 2-benzylphenyl) acetamido] thinopenicillanic acid, 6-[a-(3-methoxypheny1) acetamido]thiopenicillanic acid, 6-[a-(2-iodophenyl) acetamido] thiopenicillanic acid, 6- [oc- S-diethylaminophenyl) acetamido] thiopenicillanic acid, and 6- [a-(2,4-diisoamylpheny1) acetamido]thiopenici1lainc acid,
respectively, to produce 6- a- (4-nitrophenyl) acetamido]penici11anal,
6- [a-(4-bromophenyl) acetamido1penicillana1,
6- 0L- (4-t-butylphenyl) acetamido] penicillanal,
6- 0t- 4-trifluoromethylphenyl) acetamido penicillan a1, 6- [ac-(3-flll0l0pl16I1Y1) acetamido1penici1lanal,
6- oc- 4-sulfamy1pheny1) acetamido] penicillanal,
6- a- (Z-benzylp-henyl) acetamido] penicillanal,
6- a- 3-methoxyphenyl) acetamido] penicillanal,
6- [a- 2-i0 dophenyl acetamido] penicillanal,
6- [o.-(3-diethylaminopheny1) acetamido1penicillanal, and 6- [oc- (2,4-diisoamylphenyl acetamido1pencillanal,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system.
EXAMPLE 19 In the procedure of Example 5, the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids, i
6-( a-phenylacetamido thio-penicillanic acid,
6- a-paranitrophenylacetamido t'hiopenicillanic acid,
6- a-parachlorophenylacetamido thiopenicillanic acid,
6- oz- 4-tertiary-butylphenyl) acetamido] thiopenicillanic acid,
6-( a-ortho-tolylacetamido thiopenicillanic acid,
6- u-ortho-nitrophenylacetamido) thiopenicillanic acid,
6- a- 3,4,5 -trichloropheny1) acetamido] thiopenicillanic acid,
6- a- 3-trifluoromethylphenyl) acetamido] thiopenicillanic acid,
6- a-parabromophenylacetamido) thiopenicillanic acid,
6- (a-paraphenylphenylacetamido thiopenicillanic acid,
6- a- (4-methoxyphenyl) acetamido] thiopenicillanic acid,
6- oc- (4-cyclohexy1phenyl) acetamido] thiopenicillanic acid, I
6- a,- (2,4-dichloropheny1) acetamido] thiopenicillauic acid,
6- [oc-( 2,4-diisoamylphenyl) acetamido] thiopenicillanic acid,
6- [oc- (4-sulfamy1pheny1 acetamido] thiopenicillanic acid,
6-[ot-(2 -allyloxyphenyl) acetamido]thiopenicillanic acid,
6- a-(4-allyl-phenyl) acetamido] thiopenicillanic acid,
6- [a-(4-dimethylaminophenyl) acetamido1thiopenicillanic acid,
6- [a- (2,5dich10ropheny1) acetamido] thiopenicillanic acid,
6- oc- (2-iodopheny1 (acetarnido thiopenicillanic acid,
6- [a-(2-acetamidophenyl) acetamido] thiopenicillanic acid,
6- O6- 4-diethylaminop-henyl) acetamido] thio-penicillanic acid, and
6-[a-(3-fluorophenyl) acetamido]thiopenicillanic acid,
respectively, to produce 6- a-phenylacetamido )penicillanal,
6-( aparanitrophenylacetamido )penicillanal,
6- oc-p arachlorophenylacetamido penicillanal,
6- [a-(4-tertiary-buty1phenyl) acetamido1penicillanal, 6- a-ortho-tolylacetamido penicillanal,
6- (a-ortho-nitrophenylacetamido) penicillanal,
3-isopropyl-5-phenyl-4-isoxazolylpenicillin aldehyde, 3-tert. butyl--methyl-4-is-oxazolylpenicillin aldehyde, 3-methyl-S-p-trifluorornethylphenyla4-isoxazolylpenicillin aldehyde,
3-methyl-5-cyclohexyl-4-isoxazolylpenicillin aldehyde, 3-cyclohexyl-5-methyl-4-isoxazolylpenicillin aldehyde, 3-a-furyl-5-methyl-4-isoxazolylpenicillin aldehyde, and 3-a-thienyl-5-methyl-4-isoxazolylpenicillin aldehyde,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system;
EXAMPLE 21 In the procedure of Example 5, the potassium thiophenethicillin is replaced by an equimolar amount of the potassium salt of the acids,
respectively, to produce,
3,5-diphenyl-4-isoxazolylpenicillin aldehyde, 3-methyl-5-pheny1-4-isoxazolylpenicillin aldehyde, 3,5-dimethyl-4-isoxazolylpenioillin aldehyde, 5-benzyl-3-methyl4-isoxazolylpenicillin aldehyde, 3-methyl-S-styryl-4-isoxazolylpenicillin aldehyde, S-tert. butyl-3-phenyl-4-isoxazolylpenicillin aldehyde, 5-(2-furyl)-3-methyl-4-isoxazolylpenicillin aldehyde, 3-methyl-5-(3',5'-d-imethyl-4'-isoxazolyl)-4-isoxazolylpenicillin aldehyde, B-methyl-S-(Z-thienyl)-4-isoxazolylpenicillin aldehyde, 3-(p-chlorophenyl)-5-methyl-4-isoxazolylpenicillin aldehyde, 3- methyl-S-methylmercapto-4-isoxazolylpenicillin hyde,
alde
S-(p-chlorophenyl)-3-methyl-4-isoxazolylpenicillin aldehyde, 3 methyl S-(o-nitrophenyl)-4-isoxazolylpenicillin aldehyde, 5-isopropyl-3-methyl-4-isoxazolylpenicillin aldehyde and 5-methy1-3-(p-chlorophenyl)-4-isoxazolylpenicillin aldehyde,
respectively, each of which is isolated as a solid which is converted to an active antibacterial agent upon exposure to an oxidative enzyme system.
EXAMPLE 22 (1) 6-phenoxyacelamidopenicillanic thiol acid potassium salts.-A solution containing 35.04 g. (0.100 mole) of penicillin V (phenoxymethylpenicillin) 14.02 ml. (0.10 mole) of dry triethylamine, and 508 ml. of pure, dry DMF (dimethylformamide) was stirred and cooled to -3 C. Ethyl chloroforrnate (9.51 ml.; 0.10 mole) was added and the resulting solution was stirred at 0 for 10 minutes. A solution of 22.0 g. (0.2 mole) of sodium hydrosulfide (hydrated) in 358 ml. of pure, dry DMF was prepared by stirring at 25 C. under a nitrogen atmosphere for 10 minutes. This turbid yellow solution was added to the mixed anhydride solution all at once and the cooling bath was removed. After stirring for 10 minutes,
the reaction mixture was poured into 3.0 liters of ice water. The solution was acidified to pH 2.0 with 6 N sulfuric acid and extracted with three one-liter portions of cold ether. The combined ether solutions were washed twice with 500 ml. portions of ice water, dried very briefly over sodium sulfate and filtered. Addition of 20.5 ml. of an by weight solution of potassium-Z-ethylhexanoate (KEH) in dry n-butanol (diluted to 40 ml. with dry ether) precipitated the product, which was collected by filtration, Washed with dry ether and dried, yield 32.7 g. (81%). The product was recrystallized by dissolving in a minimum of cold water (ca. 40 ml.), adding ca. 500 ml. of n-butanol, and removing the water-butanol azeotrope in vacuo at 33 C. until the solution was entirely dry. The purified yield was 28.2 g. (70%). The infrared absorption spectrum was identical to that of the original sample of this material for which excellent analytical data had already been obtained.
(2) 3 (1,3 diphenyl-2-imidaz0lidinyl)-2,2-dimethyl- 6-phenoxyacetamidopenam.-A suspension of 52.0 g. of Raney active nickel No. 28 (washed by decantation three times with ml. portions of absolute ethanol and then four times with 105 ml. portions of THF) in 160 ml. of tetrahydrofuran (TI-1 F) containing 3.20 ml. of glacial acetic acid, 5.00 g. (0.0235 mole) of N,N'-diphenylethylenediamine, and 15.0 ml. of water was cooled to 0 C. To this vigorously stirred solution was added a solution of 6.35 g. (0.0157 mole) of 6-phenoxyacetamidopenicillanic thiol acid potassium salt in 60 ml. of THF containnig 6.25 ml. of water. An eifervesce was noted, which subsided after about one minute. The mixture was stirred at 60 C. for 15 minutes. The catalyst was then filtered oit over diatomaceous earth (Supercel), and the nickel was washed on the filter with five 50 ml. portions of THF. The solvent was removed in vacuo from the pale yellow filtrate, and the residue was flash-distilled with two 100 ml. portions of dry ethyl acetate at 33 C. to remove traces of moisture. Five hundred ml. of anhydrous ether was added and the turbid solution was refluxed for onehalf hour. During this period of reflux, a white, crystalline precipitate formed. At the end of the reflux period, the warm solution was filtered, and the solids and filtrate were worked up separately.
The solids were boiled briefly with 100 ml. of ethyl acetate and the hot mixture was filtered. On thorough cooling, the filtrate yielded 2.70 g. of white crystals, M.P. 174-175 C. Then the filtrate from this portion of the product was freed of solvent in vacuo and the residue was treated with 15 ml. of dry ether, a further quantity of the product, 1.42 g., was obtained as a highly crystalline but slightly tan solid, M.P. l67l70 C.
The filtrate was extracted with 20 ml. of 10% phosphoric acid, which caused N,'Ndiphenylethylenediarnine phosphate to crystallize. It was filtered off, the aqueous phase was separated, and the other solution was again extracted with 20 ml. of 10 phosphoric acid and washed with three 100 ml. portions of water. It was dried over sodium sulfate, filtered, flashed dry and flashed with ethyl acetate. The addition of 10 ml. of dry ether to the small residue caused 0.25 g. of product to crystallize, M.P. 164168 C.
The total yield was 4.37 g. (52.5%) of the title compound.
(3) 6 phenoxyacetamidopenicillanaL-A solution of 10.5 g. of pure 3(1,3-diphenyl-2-imiclazolidinyl)-2,2-dimethyl-6-phenoxyacetamidopenam (0.189 mole) was prepared in 98.5 ml. dry acetone, 98.5 .ml. of methylene chloride and 298 ml. of anhydrous ether. To this solution was added a solution of 7.11 g. (0.374 mole) of ptoluenesulfonic acid monohydrate in a mixture of 50 ml. of CH Cl 50 ml. of dry acetone, ml. of anhydrous ether, and 1.3 ml. of water. A heavy precipitate formed instantly, and the slurry was stirred mechanically at 25 for /2 hour. -It was then diluted with 1.1 liter of ether and filtered over Supercel. The clear filtrate was ex- 'acted with two 100 ml. portions of water, dried over soium sulfate, filtered and flashed dry at 33 C. Fifty ml. f dry ether was added and the solution cooled in ice for t least 15 minutes. The product crystallized and filtraon yielded 2.08 g. of white needles, M.P. 106108 C. he filtrate was allowed to evaporate with 1.5 ml. of H dded at 20 C. for 24 hours. The crystals were filtered nd the yield was 3.90 g. of white needles, M.P. 105104 2. The total yield was 5.98 g. (94.5%).
The infrared absorption spectrum was quite sharp, howing NH 3380 c-m.- fl-lactam 1780, aldehyde 1740, mide 1670 and 1525 cmf All of the bands noted were -f equal intensity. The aldehyde may be recrystallized cut at 33 C. Water (a few ml.) and ordinary ether cetate (three parts per 100 ml.) and flashing off the solent at 33 C. Water (a few ml.) and ordinary ether one part per 100 ml.) are added and the resulting soluion is allowed to stand at 20 C. for 24 hours. The prodict will slowly crystallize.
Analysis.-Calcd for C H N O S- /2H O: C, 55.96; i, 5.57; N, 8.16; S, 9.34. Found: C, 55.40, 55.25; H, L63, 5.72; N, 8.08; S, 9.12.
(4) 2,2 dimethyl-3-acetoxymethylene-6-phenoxyacefzmid0penam.A mixture of 2.07 g. (0.00604 mole) of )BIllCilllIl V aldehyde hemihydrate, 200 ml. of acetic aniydride, and 3.0 ml. of pyridine was stirred at 25 under 1 nitrogen atmosphere for 16 hours. The acetic anrydride and excess pyridine were removed in vacuo at 50 C. The dark residue was dissolved as far as possible n 150 ml. of carbon tetrachloride and the solution was lined with ca. 4 g. of decolorizing charcoal for 45 minites. The solution was filtered over Supercel and the iltrate was flashed dry at 50 C. In order to codistill ost of the remaining traces of acetic anhydride, 75 ml. of carbon tetrachloride was added and the flashing was repeated. The light orange residue was dissolved in a few ml. of methylene chloride, flashed to an oil and dried to a friable fluff in high vacuum at 25 C. The yield was 1.11 g. (49%) of light orange foam. The infra-red spectrum (5% in CCl shows NH at 3400 cmr ,8- lactam 1800, enol acetate carbonyl 176 2, amide 1700 and 1510 and COC bands at 1230, 1190, and 1090 crnf A weak band at 1640 cm.- is ascribed to The NMR spectrum is in full agreement with the enol acetate structure and shows further that the product is a mixture of approximately equal amounts of cis and trans isomers.
Analysis.Calcd for C H N O S: C, 57.43; H, 5.63. Found: C, 57.20; H, 5.37.
PREPARATION OF STARTING MATERIALS PENICILLIN THI'OACIDS Another series of starting materials used to prepare the compounds of the present invention comprises the series of compounds of the formula wherein R is amino, acylarnino, ibenzyloxycarbonylamino, phthalimido or tritylamino and, particularly, the so-called penicillin thioacids or thiopenicillins of the formula S CH3 ll in which R represents the side chain of any of the known penicillins other than those containing a group which is altered by reaction with Raney nickel above 0 C. These compounds are usually used in the form of salts, e.g. sodium, potassium.
These compounds exist in tantomeric forms, i.e. the -COSH group may be in the thiol form as or in the thione form as Such thiopenicillins are prepared from penicillins by the methods described below or in US. Patent 2,751,378. In general, to prepare a thiopenicillin, an active acylating derivative of the starting penicillin such as an anhydride or a mixed anhydride (such as the mixed anhydride with a lower alkyl ester of ethoxy-or isobutoxy-carbonic acid) or an acid chloride is prepared and reacted with a source of sulfhydryl groups, e.g. hydrogen sulfide or sodium hydrosulfide or potassium hydrosulfide.
Additional examples of the preparation of thiopenicillins are as follows:
Preparation 11.- Preparati0n 0 potassium 6-(DL-uph enoxy pro pionamido) thio p em'cil lanate Dilute sulfuric acid is added to a solution of potassium 6-(DL-a-phenoxypropionamido)penicillanate (10 gm., 0.025 mole), dissolved in water ml.) until a pH of 2 is attained. The penicillin acid is extracted from this solution into ether, washed with water, and dried over anhydrous magnesium sulfate. The ether is evaporated under reduced pressure to leave a residual oil which is azeotroped three times with ethyl acetate to remove all traces of water. The residue is dissolved in dimethylformamide ml.) and cooled to 5 C. in an ice bath. 2,6-lutidine (2.7 gm., 0.0254 mole) is added, followed by the dropwise addition of ethyl chloroformate (2.7 gm., 0.0254 mole). The mixture is stirred for 15 minutes and a suspension of sodium hydrosulfide (5.6 gm., 0.06 mole) in dimethylformamide (100 ml.) is added all at once. The solution is stirred for one hour, and turns dark brown. The solution is poured into water (one liter) and acidified to pH 2 with dilute sulfuric acid. The 6 (DL-oc phenoxypropionamido)thiopenicillanic acid is extracted into ether, washed with water, and dried over anhydrous magnesium sulfate. Potassium 2-ethy1- hexanoate (5 gm.) is added and the crystalline precipitate is collected and weighs 4.5 gm. Recrystallization from water and acetone yields 1.1 gm. of the product, potassium 6 (DL a phenoxypropionamido)thiopenicil lanat'e, as colorless plates which are found to contain the B-lactam ring as shown by infrared analysis, to have a melting point of greater than 240 C. with decomposition and the following elemental anaylsis:
Calculated for C17H19N2O4S2K'1/2H2OZ C, H, 4.74%; S, 14.9%. Found: C, 48.2%; H, 4.52%; S, 13.8%.
Preparation 12 In procedure of Preparation 11, the potassium 6- (u-phenoxypropionamido)pencillanate is replaced by 0.025 mole of the potassium salt of the acids 6- a- 2-chlorophenoxy propionamido] penicillanic acid,
6 [on (4 sulfamylphenoxy) n butyram-ido]penicil lanic acid,
6 [a (3,4 dimethoxyphenoxy) n pentanoamido] penicillanic acid,
6 [a (3 methylphenoxy)isovaleramido]penicillanic acid,
6 [0: (4 methylthiophenoxy)propionamidoJpen-icillanic acid,
6 d m thylaminophenoxy) n hexanoamido] enicillanic acid,
6 [a (2 inethoxyphenoxy) -1 n decanoamido1penici-llanic acid,
6 [a (2,4 dichlorophenoxy)phenylacetamidoJpenicillanic acid,
6 [a (2 nitrophenoxy) ,8 phenylpropionamido] penicillanic acid,
6 [a (2 a-cetamidophenoxy) 'y phenylbutyramido] penicillanic acid,
6 [a (2,4 dimethylphenoxy) n butyramido1penicillanic acid,
6 [a (4 isopropylphenoxy)propionamido]penicillanic acid,
6 [a (3 bromophenoxy) n butyramido]penicillanic acid,
6 (2 iodophenoxy)phenylacetamido]penicillanic aci 6 [c (2 diethylaminophenoxy)isovaleramido]penicillanic acid,
6 [0c (3,5 dichlorophenoxy)isohexanoamido1penicillanic acid,
6 [0c (4 cyclohexyphenoxy)propionamido1penicillanic acid,
6 [a phenoxy isovalerarnido]penicillanic acid,
6-[a-phenoxy-n-deoanoamido] penicillanic acid,
6-[ot-phenoXy-y-phenylb utyramido]penicillanic acid,
5 [a (2 benzylphenoxy) n butyramido]penicillanic acid,
6 [a 4 (Z-trifluoromethylphenoxy)propionamido]penicillanic acid, and
6- a- (4-fluorophenoxy propiona-mido] penicillanic acid,
to produce the potassium salts of 6-[ a-(2-chlorophenoxy) propionaniido] thiopenicillanic acid,
6- [a- (4-sulfamylphenoxy) n-butyramidoJthiopenicillanic acid,
6- [a- 3,4-dimethoxyphenoxy) -n-pentanoamido] thiopenicillanic acid,
6- [a- 3 -met'nylphenoxy isovalerarnido] thiopenicillanic acid,
6-[a-(4-methylthiophenoxy)propionamido] thiopenicillanic acid,
6-[a(4-din1etnylarnmophenoxy) -nhexanoamido] thlopenicillanic acid,
6- ot- (Z-metnoxyphenoxy -n-decanoa-mido] thiopenicillanic acid,
6-[a-(2,4-dichlorophenoxy)phenylacetamido] thiopenicill-anic acid,
6-[tx-(Z-nitr-ophenoxy)p phenylpropionamido] thiopenicillanic acid,
6- a- (Z-acetamidophenoxy) -7-phenylbutyramido] tniopenicillanic acid,
6-[ot-(2,4-dimethylphenoxy)-n-butyramido] thiopenicillanic acid,
6-[ot-(4-isopropylphenoxy)propionarnido1thiopenicillanic acid,
6- a- 3-bromophenoxy) -n-butyramido] thiopenicillanic acid,
6- a- (2-iodophenoxy phenyla-cetamido] thiopenicillanic acid,
6- 0t- (2-diethylaminophenoxy isovaleramido] thiopenicillanic acid,
6-[a-(3,5-dichloro-phenoxy)is-ohexanoamido1 thiopenicillani-c acid,
6- a- (4-cyclohexylphenoxy) propionamido] thiopcnicillanic acid,
6-[a-phenoXy-isovalerarnido1thiopenicillanic acid,
6-[owphenoxy-n-decanoamido1thiopenicillanic acid,
6-[a-phenoxy-v phenylbutyramid-o] thiopenicil'lanic acid,
6- a( Z-benzylphenoxy -n-butyramido] thiopenicillanic acid 6-[a-(2-trifiuoromethylphenoxy)propionamido]thiopencillanic acid, and
6-[w (4-fluorophenoxy)propionamido]thiopenicillanic acid,
.34 respectively, each of which is isolated as its solid, waterso'luble potassium salt.
Preparation 13.-Preparati0n of potassium 6-(D-uphenoxypropionamido)thiopcnicillanate Dilute sulfuric acid is added to a solution of potassium 6-'(D a phenoxypropionamido)penicillanate (5 gm.,
0.0125 mole) in water (150 ml.) until a pH of 2 is attained. The penicillin acid is extracted from this solution into ethyl acetate (200 ml.), washed with water, and dried over anhydrous magnesium sulfate. The ethyl acetate is evaporated at 35 C. The residue is dissolved in dimethylforrnamide (150 ml.) and cooled to 5 C. in an ice bath. 2,6-lutidine (1.33 gm., 0.0125 mole) is added, followed by the dropwise addition of ethyl chloroformate (1.33 gm., 0.0125 mole). The mixture is stirred for 15 minutes and a suspension of sodium hydrosulfide (2.5 gm., 0.0447 mole) in dimethylformamide :ml.) is added all at once. The solution is stirred for 20 minutes and then poured into water (one liter) and acidified to pH 2 with dilute sulfuric acid. The 6-(D-ct-Ph6ll0XY- propionamido)thiopenicillani-c acid that forms is extracted into ether, washed with Water, and dried over anhydrous magnesium sulfate. Potassium Z-ethylhexanoate (3 gm.) is added and the crystalline precipitate is collected. Recrystallization from water and n-butanol yields 1.7 gm. of the product, potassium 6-(D'a-phenoxypropionamido)thiopenicillan-ate, which is found to contain the fi-lactam ring as shown by infrared analysis, to have a melting point of greater than 195 C. with decomposition and the following elemental analysis:
Calculated for C H N O S K: C, 48.78%; H, 4.58%; N, 6.7%; S, 15.4%. Found: C, 49.25%; H, 4.68%; N, 6.84%; S, 14.52%.
Preparation 14.---Preparation of potassium 6-(L-aphenoxypropionamido)thiopenicillanate Dilute sulfuric acid is added to a solution of potassium 6 (L-u-phenoxypropionamido)penicillanate (5 gm., 0.0125 mole) in water ml.) until a pH of 2 is attained. The penicillin acid is extracted from this solution into ethyl acetate (200 ml.), washed with water, and dried over anhydrous magnesium sulfate. The ethyl acetate is evaporated at 35 C. The residue is dissolved in dimethylformamide (150 ml.) and cooled to 5 C. in an ice bath. 2,6-lutidine (1.33 gm., 0.0125 mole) is added, followed by the dropwlse addition of ethyl chloroformate (1.33 gm., 0.0125 mole). The mixture is stirred for 15 minutes and'a suspension of sodium hydrosulfide (2.5 gm., 0.0447 mole) in dimethyl'formamide (100 ml.) is added all at once. The solution is stirred for 20 minutes and then poured into water (one liter) and acidified to pH 2 with dilute sulfuric acid. The 6-(L-a-phenoxypropionamido)thiopenicillanic acid is extracted into ether, washed with water, and dried over anhydrous magnesium sulfate. Potassium 2-ethylhexan0ate (3 gm.) is added and the crystalline precipitate is collected. Recrystallization from water and n-butan-ol yields 2.5 gm. of the product, potassium 6-(L-ot-phenoxypropionamido)thiopenicillanate which is found to have a melting point of greater than 215 C. with decomposition and the following elemental analysis:
Calculated for C17H19N204S2K1 'C, H, N, 6.7%; S, 15.4%. Found: C, 50.22%; H, 4.98%; N, 6.77%; S, 14.18%.
Preparation 15.--Preparati0n of potassiumo-(aisopropyl-u-phenoxyacetamido)thiopenicillanate Dilute sulfuric acid is added to a solution of potassium 6-(a-isopropyl-a-phenoxyacetamido)thiopenicillanate (5.3 gm., 0.0125 mole) in water (150 ml.) until a pH of 2 is attained. The penicillin acid is extracted from this solution into ethyl acetate (200 m1.) washed with water, and dried over anhydrous magnesium sulfate. The ethyl acetate is evaporated at 35 C. The residue is disolved in dimethylformamide (150 ml.) and cooled to i C. in an ice bath. 2,6-lutidine (1.33 gm., 0.0125 mole) s added, followed by the dropwise addition of ethyl chlo- 'oformate (1.33 gm., 0.0125 mole). The mixture is :tirred for 15 minutes and a suspension of sodium hydro- ;ulfide (2.5 gm., 0.0447 mole) in dimethylform-amide 1100 ml.) is added all at once. The solution is stirred for 20 minutes and then poured into water (one liter) and tcidified to pH 2 with dilute sulfuric acid. The 6-(asopropyl-a phenoxyacetamido)thiopenicillanic acid is ex- :racted into ether, Washed with water, and dried over anhydrous magnesium sulfate. Potassium Z-ethylhexanoate (3 gm.) is added and the crystalline precipitate is :ollected. Recrystallization from ethyl acetate and Skellysolve yields 2.3 gm. of the product potassium 6-(alsopropyl-a-phenoxyacetamido)thiopenicillanate, which is found to contain the B-lactam ring as shown 'by infrared analysis and to have a melting point of greater than 170 C. with decomposition.
Preparation 1 6 .--Preparation of potassium 6-[L( -ocphenoxybutyram ido] thio p enici l lanate Potassium 6- [L( )-a-phenoxybutyramido]penicillanate (2.08 gm., 0.005 mole) is dissolved in water (30 ml.) and layered with ethyl acetate. After cooling to 5 C., dilute phosphoric acid (40%) is added until a pH of 2 is attained. The penicillin acid is extracted into the ethyl acetate and a further extraction with fresh ethyl acetate is made. The combined extracts are washed with ice- Water, dried over anhydrous magnesium sulfate, and evaporated to dryness. The residue is dissolved in dimethylformamide (30 ml.) and cooled to C. in an ice bath. 2,6-lutidine (0.53 gm., 0.005 mole) is added, followed by the dropwise addition of ethyl chloroformate (0.52 gm., 0.005 mole) forming a precipitate. The mixture is stirred for l0'mirrutes at 0 C. and a solution of sodium hydrosulfide trihydrate (1.1 gm., 0.01 mole) in dimethylformamide (30 m1.) is added in one portion. The resulting green solution is stirred for 25 minutes and poured into a precooled (10 C.) mixture of water (150 ml.), acidified to pH 1.5 with dilute phosphoric acid (40%) and benzene (100 ml.) with vigorous stirring. The 6- [L()-a-phenoxybutyramido]thiopenicillanic acid is extracted into benzene and a further extraction with fresh benzene is made. The combined extracts are washed and dried. Potassium 2-ethylhexanoate (0.005 mole) is added as a 50% solution of potassium Z-ethylhexanoate in butanol. Skellysolve B (a petroleum ether fraction having a boiling point range of from about 60 to about 68 C. consisting essentially of n-hexane) is added to the solution with cooling and shaking until the solution becomes cloudy, and an oil separates. The solution is decanted and flashed to dryness. The residue is triturated with ether, and a white solid forms which is removed by filtration, washed with dry ether, and dried in vacuo. The product, potassium 6- [L( )-a-phenoxybutyramido] thiopenicillanate is found to weigh 2 grams, to have a melting point of 145 149 C. with decomposition and the structure is confirmed by infrared analysis.
Preparation 17 In the-procedure of Preparation 11, the potassium 6- (a-phenoxypropionarnido)penicillanate is replaced by 0.025 mole of the potassium salt of 6- a-phenoxy-a-benzylpropionamido penicillanic acid,
6- a-phen'ylthio-a-benzylpropionamido penicillanic acid,
6- a-phenoxy-a-methylpropionamido penicillanic acid,
6- a-phenylthio-a-methylpropionamido penicillanic acid,
6- [a- 2,4-dichl orophenoxy) -a-ethylpropionamido] penicillanic acid,
6- a- Z-chlorophenoxy) -a-methylbutyramido] penicillanic acid,
6 a- (4-nitrophenoxy -a-methyl-n'-butyramido] penicillanic acid,
6- a- 4-b romophenoxy) -a-phenyl-n-valeramido] penicillanic acid,
6-[a-(4-t-butylpl1enoxy) -a-benzyl-n-butyramido] penicillanic acid,
6- [oc- (4-trifluoromethylphenoxy -a-phenyl-n-butyramido] penicillanic acid,
6- a- 3 -fluorophenoxy -a-benzyl-.n-valeramido] penicillanic acid,
6- oc- 4-sulfamylphenoxy) -u-methylpr-opionamido] penicillanic acid,
6- a- Z-benzylphenoxy) -a-methyl-n-butyramido] penicillanic acid,
6 [a- 3-methoxyphenoxy) -a-methyldecanoamido] penicillanic acid,
6- [oc- Z-iodophenoxy) -a-phenylpropionamido] penicillanic acid,
6 [a- B-diethylaminophenoxy) -a-methyl-n-butyramido] penicillanic acid, and
6- rx- 2,4-diisoamylphenoxy) -a-methyl-n-butyramido] penicillanic acid,
to produce the potassium salts of 6- a-phenoxy-a-benzylpropionamido thiopenicillanic acid,
6 a-phenylthio-a-benzylpropionamido) thiopenicillanic acid,
6- a-phenoxy-a-methylp ropionamido thiopenicillanic acid,
6- u-phenylthio-a-methylpropionamido thiopenicillanic acid,
6- oc- 2,4-dichlorophen0xy a-ethylpropion-arnido] thiopenicillanic acid,
6- [a- 2-ch1orop-henoxy) -a-methylbutyramido] thiopenicillanic acid,
6- a- 4-nitrophenoxy -a-methyl-n-butyramido] thiopenicillanic acid,
6- a- (4-bromophenoxy) -a-phenyl-n-valeramido] thiopenicillanic acid,
6- a- 4-t-butylphenoxy) -a-benzyl-n-butyramido] thiopenicillanic acid,
6- [a- 4-trifiuoromethylphenoxy) -a-phenyl-n-butyramido] thiopenicillanic acid,
6- [a- 3-fiuorophenoxy) -a-benzyl-n-valeramido] thiopenicillanic acid,
6-[a-4-sulfarnylphenoxy)-a-methylpropionamido] thiopenicillanic acid,
6- [a- Z-benzylphenoxy -a-methyl-n-butyramido] thiopenicillanic acid,
6- a- 3-rnethoxyphenoxy) -a-methyldecanoamido] thiopenicillanic acid,
6- a- 2-iodophenoxy) -a-phenylpropionamido] thiopenicillanic acid,
6- cx- 3-di-ethylaminophenoxy -a-methyl-n-butyramido] thiopenicillanic acid, and
6-[a-(2,4-diisoamylphenoxy)-a-methyl-n-butyramido] thiopenicillanic acid,
respectively, each of which is isolated as its solid, watersoluble potassium salt.
Preparation 18 In the procedure of Preparation 11, the potassium 6- (a-phenoxypropionamido)penicillanic acid is replaced by 0.025 mole of the potassium salt of 6- a-phenylthiopropionamido penicillanic acid, 6- a-p aranitrophenylthiopropionamido penicillanic acid, 6- a-parachlorophenylthiopropionamido penicillanic acid, 6- a-phenylthiobutyrarnido penicillanic acid, 6-( a-phenylthiocaproamido penicillanic acid, 6- a-phenylthioisovaleramido penicillanic acid, 6- a- (4-t-butylphenylthio propionamido] penicillanic acid, 6- wortho-tolylthiopropionamido penicillanic acid, 6- a-orth o-nitrophenylthiopropionamido) penicillanic acid, 6- a-parachlorophenylthiobutyramido) penicillanic acid,
6-[tx-(3,4,S-trichlorophenylthio)propionarnido] penicillanic acid,
6- oc- 3-trifluoromethylphenylthio) butyramido] penicillanic acid,
6 a-p arabromophenylthioisovaleramido) penicillanic acid,
6- a-paraphenylphenylthiopropionamid'o) penicillanic acid,
6- [a- (4-methoxyphenylthio) caproamido] penicillanic acid,
6- oc- 4-cyclohexylphenylthio) butyramido] penicillanic acid,
6-(a-phenylthio-wcyclohexylacetarnido) penicillanic acid,
6-(a-phenylthio-a-cyclopentylacetamido) penicillanic acid,
6-[a-(2,4-dichlorophenylthio)caproamido] penicillanic acid,
6-[u-(2,4-diisoarny1phenylthio)propionamido] penicillanic acid,
6-[a-(4-benzylphenylthio)propionamido] penicillanic acid,
6-[w(4-sulfarnylphenylthio)butyramido] penicillanic acid,
6-[u-(2-allyloxyphenylthio)propionamido] penicillanic acid,
6-[a-(4-allylphenylthio)isovalerarnido] penicillanic acid,
6-[w(4-dimethylarninophenylthio)propionamido] penicillanic acid,
6-[a-(2,5-di-chlorophenylthio)butyramido] penicillanic acid,
6- a- 2-iodopheny1thio propionamido] penicillanic acid,
6-[a-(Z-acetamidophenylthio)propionamido] penicillanic acid,
6- [t- 4-diethylaminophenylthio propionamido] penicillanic acid, and
6- a- 3-fiuorophenylthio) butyramido] penicillanic acid,
to produce the potassium salts of 6-(oephenylthiopropionamido)thiopenicillanic acid,
6-(a-paranitrophenylthiopropionamido) thiopenicillanic acid,
6-( oc-parachlorophenylthiopropionamindo) thiopenicillanic acid,
6-( a-phenylthi obutyramido) tbiopenicillanic acid,
6-(ot-pheny1thiocaproamido)thiopenicillanic acid,
6 (a-phenylthioisovalerami do)thiopenicil1anic acid,
6-[a-(4-t-butylphenylthio)propionamido] thiopenicillanic acid,
6- [a-ortho-tolylthiopropionamido] thiopeni-cillanic acid,
6-(a-ortho-nitrophenylthiopropionarnido) thiopenicillanic acid,
6-( wparachloro-phenylthiobutyramido) thiopenicillanic acid,
6- [a- (3 ,4,5-trichlorophenylthio) propionamido] thiopenicillanic acid,
6-[01-(3 -trifluoromethylphenylthio) butyramido] thiopeni-cillanic acid,
6-(a-parabromophenylthioisovaleramido) thiopenicillanic acid,
6-(a-paraphenylphenylthiopropionarnido) thiopenicillanic acid,
6- [a- (4-methoxyphenylthio cap-roamido] thiopenicillanic acid,
6- O!.- (4-cyclohexylphenylthio butyramido] thiopenicillanic acid,
6 a-phenylthioa-cyclohexylacetamido) thiopenicillanic acid,
6-(oc-phenylthioa-cyc1opentylacetamido) thiopenicillanic acid,
6-[a-(ZA-dichlQrophenyIthio)capro-amido] thiopenicillanic acid,
33 6-[a-(2,4-diisoamylphenylthio)propionamido] thiopenicillanic acid, 6- a- (4-benzylphenylthio propionamido] thiopenicillanic acid, G-[a-(4-sulfalmylphenylthio)butyramido] thiopeni-cillanic acid, 6-[a-(2-allyloxyphenylthio)propionarnido] thiopenicillanic acid, 6-[oz-(4-allylphenylthio)isovaleramido] thiopeni-cillanic acid, 6-[a-(4-dimethylaminophenylthio)propionamido] thiopenicillanic acid, 6-[oz-(2,5-dichlorophenylthio)butyramido] thiopenicillanic acid, 6-[u-(2-iodophenylthio)propionamido] thiopenicillanic acid, 6-[ot-(Z-acetamidophenylthio)propionamido] thiopenicillanic acid, 6-[a-(4-diethylarninophenylthio)propi onamido] thiopenicillanic acid, and 6-[a-(3-fluorophenylthio)butyramidoj] thiopenicillanic acid, respectively, each of which is isolated as its solid water,- soluble potassium salt.
Preparation 19 In the procedure of Preparation 16 the 2.08 gm. of potassium 6-[L() a phenoxybutyramidojpenicillanic acid is replaced by 8.32 grams of the same acid to produce in the first crop 4.0 grams of the desired product, potassium 6 [L( oz phenoxybutyramido]thiopenicillanic acid in crystalline form, having purity of about (as shown by NMR studies), a specific rotation of 177 (C.= /2 H 0) and a melting point range of from 155 to 160 C. with decomposition.
Preparation 20.Preparati0n of potassium 6- [D -ecplzenoxybulyramido] thiopenicillanale Potassium 6-[D( -0t phenoxybutyramido]penicillanate (3.0 gm.) is dissolved in water (30 ml.) and layered with ethyl acetate. After cooling to 5 C.., dilute phosphoric acid (40%) is added until a pH of 2 is attained. The penicillin acid is extracted into the ethyl acetate and a further extraction with fresh ethyl acetate is made. The combined extracts are washed with ice-water, dried over anhydrous magnesium sulfate, and evaporated to dryness. The residue is dissolved in dimethylformanide (30 ml.) and cooled to 0 C. in an ice bath. 2,6-lutidine (0.53 gm., 0.005 mole) is added, followed by the drop Wise addition of ethyl chloroforrnate (0.52 gm., 0.005 mole) forming a precipitate. The mixture is stirred for 10 minutes at 0 C. and a solution of sodium hydrosulfide trihydrate (1.1 gm., 0.01 mole) in dimethylformamide (30 ml.) is added in one portion. The resulting green solution is stirred for 25 minutes and poured into a precooled (5-l0 C.) mixture of water (150 1111.), acidified to pH 1.5 with dilute phosphoric acid (40%) and benzene ml.) with vigorous stirring. The t 6 [D(+) a phenoxybutyrarnido]thiopenicillanic acid is extracted into benzene and a further extraction with fresh benzene is made. The combined extracts are washed and dried. Potassium Z-ethylhexanoate (0.005 mole) is added as a 50% solution of potassium 2- ethylhexanoate in butanol. Skellysolve B is added to the solution with cooling and shaking until the solution becomes cloudy, and an oil separates. The solution is decanted and flashed to dryness. The residue is triturated with ether, and a white solid forms which is removed by filtration, washed with dry ether, dried in vacuo and found to weigh 2.5 gm. The product, potassium 6 [D(+) a phenoxybutyramido]t'hiopenicillanate, is found to have a melting point of -l45 C. with decomposition, and the structure is confirmed by infrared analysis.
39 Preparation 21 .-Preparation f potassium 6- [D,L-OL- phenoxypropionamido] thiopenicillanafe Potassium 6 [D,L-ot-phenoxypropionamido]penicillante (30.0 gm.) is dissolved in a mixture of 100 ml. ice 'ater and 100 ml. trichlorethylene. After cooling the iixture to C., 6 N hydrochloric acid is added with :irring until a pH of 2 is attained. The trichlorethylene base is separated and maintained at 5 C. The aqueous base is extracted again with 50 ml. of trichlorethylene and 1e trichlorethylene extracts are combined, mixed with 40 m. of anhydrous magnesium sulfate and stirred in an ice ath for minutes. The combined extracts are then fil- :red and the filter cake is washed with 50. ml. of trihlorethylene. The filtrates are placed together with 250 31. of dimethylformamide in a one-liter 3-necl ed flask vith a drying tube vent and cooled to 3 C. in an icevcetone bath. To the chilled filtrates is then added 10.2 n1. of 2,6-lutidine and then ethylchloroformate (8.7 ml.) W61 a five-minute period after which the resulting soluion of mixed anhydride is stirred in the ice bath for 30 ninutes. An anhydrous solution of 16.8 gm. of sodium iydrosulfide trihydrate in 150 ml. of dimethylformamide s mixed with the mixed anhydride solution over a period )f five minutes and the resulting slurry is stirred for one mm at 0 C. The slurry is then slowly decanted into 1500 ml. of ice water having a pH of 1.9. The pH of the nixture is maintained at about 2 by adjustment With 6 N iydrochloric acid. The aqueous solution of 6-[D,L-a- Jhenoxypropionamido]thiopenicillanic acid is extracted 1nd re-extracted with trichlorethylene. The combined :richl-orethylene extracts are dried over anhydrous magnesium sulfate in an ice bath and placed under vacuum for 30 minutes to remove hydrogen sulfide. The magnesium sulfate is then filtered off and washed on the filter with trichlorethylene. To the combined filtrates is added 70 ml. of a 22% solution of potassium ethylhexanoate in methyl isobutyl ketone whereupon the product, potassium 6- [D,L-ot-phenoxypropionamido] thiopenicillanate, crystallizes out of solution. The slurry is stirred for 20 minutes at room temperature and thereafter in an ice bath for 75 minutes While the product crystallizes. The product is collected by filtration, washed with trichlorethylene, vacuum dried over P 0 for 18 hours and thereafter found to weigh 23.9 gm.
' Preparation 22.Preparati0n of potassium 6- [D,L-u
phenoxybutyramido] thiopenicillanate Potassium 6- [D,L-ot-phenoxybutyramido] penicillanate (30.0 gm.) is dissolved in a mixture of 100 ml. ice Water and 100 ml. trichlorethylene. After cooling the mixture to 5 C., 6 N hydrochloric acid is added with stirring until a pH of 2 is attained. The trichlorethylene phase is separated and maintained at 5 C. The aqueous phase is extracted again with 50 ml. of trichlorethylene and the trichlorethylene extracts are combined, mixed with 40 gm. of anhydrous magnesium sulfate and stirred in an ice bath for 30 minutes. The combined extracts are then filtered, and the filter cake is washed with ml. trichlorethylene. The filtrates are placed together with 250 ml. dimethylformamide in a one-liter 3-necked flask with a drying tube vent and cooled to -3 C. in an ice-acetone bath. To the chiiled filtrates is then added 10.2 ml. 2,6-lutidine and then ethylchloroformate, 8.7 ml. over a five-minute period after which the resulting solution of mixed anhydride is stirred in the ice bath for 30 minutes. An anhydrous solution of 16.8 gm. of sodium hydrosulfide trihydrate in 150 ml. of dimethylformamide is mixed with the mixed anhydride solution over a period of five minutes and the resulting slurry is stirred for one hour at 0 C. The slurry is then slowly decanted into 1500 ml. of ice water having a pH of 1.9. The pH of the mixture. is maintained at about two by adjustment with 6 N; hydrochloric acid. The aqueous solution of 6-[D,L-a-phenoxybutyramido] thiopenicillanic acid is extracted with trichlorethylene. The combined trichlorethylene extracts are dried over anhydrous magnesium sulfate on an ice bath and placed under vacuum for 30 minutes to remove hydrogen sulfide. The magnesium sulfate is then filtered 01f and washed on the filter with trichlorethylene. To the combined filtrates is added ml. of a 22% solution of potassium ethylhexanoate in methyl isobutyl ketone whereupon the product, potassium 6-[D,L-ot-phenoxybutyramido]thiopenicil lanate, crystallizes out of solution. The slurry is stirred for 20 minutes at room temperature and thereafter in an ice bath for minutes While the product crystallizes. The product is collected by filtration, washed with trichlorethylene, vacuum-dried over P 0 for 18 hours, and thereafter the structure is confirmed by nuclear magnetic resonance and infrared absorption data.
Preparation 23.-Preparati0n of 6-aminothiopenicillanic acid A fermentation broth is prepared by the fermentation of Escherichia coli under submerged aerobic conditions according to conventional procedures and found to contain 5,270 penicillin amidase units per ml.
A solution of 2.5 gm. calcium nitrate dihydrate dissolved in 6 ml. of water is added to one liter of such fermentation broth. After mixing, the broth is filtered, and the filtered mat washed with 150 ml. water. The filtered mat is then suspended in 200 ml. of water to which is added 5 ml. of toluene, and the suspension is stirred for three hours. After stirring, the suspension is filtered and the collected solid materials washed with ml. of water. The filtrate is stirred with 1 gm. activated carbon (Darco KB) and 1.2 ml. of Quaternary Ammonium Salt Mixture No. I. Quaternary Ammonium Salt Mixture No. I is commercially available from Armour & Company of Chicago, Ill., under the trademark of Arquad 16-50 and is a liquid quaternary ammonium salt mixture containing, by weight, about 45% hexadecyltrirnethylammonium chloride, about 3% octadecyltrimethylammonium chloride, about 2% octadecenyltrimethylammonium chloride, about 35% isopropanol, about 14% water, and about 1% sodium chloride.
After thorough mixing, all solid materials are remove from the mixture by filtration, and there is obtained 280 ml. of an essentially pure aqueous solution of penicillin amidase assaying 1950 penicillin amidase units per ml. and containing about 10% of the penicillin amidase originally present in the broth.
Benzylthiopenicillin (8 gm.) is prepared by the proce-' dure described in United States Patent No. 2,751,378 and added to 400 ml. of penicillin amidase solution, prepared according to the procedure described above. This suspension is maintained at pH 8.0 and 35 C. for four hours, during which time the enzyme brings about the enzymatic hydrolysis of the benzylthiopenicillin. The benzylthiopencillin and phenylacetate (produced during the hydrolysis) are removed at pH 2 and 5 C. with methyl isobutyl ketone. After extraction, a portion of the liquor (250 ml.) is adjusted to pH 7 with sodium hydroxide and vacuum concentrated to 20 ml. The concentrate is adjusted to pH 4 with hydrochloric acid (6 N) and cooled to 5 C. and allowed to stand for 20 hours at 5 C. during which time crystallization occurs. The crystals are filtered, washed with water (10 ml.) and then dry acetone. The 6-aminothiopenicillanic acid is recovered, weighs 0.29 gm. and the presence of the o H -CSH group and ,B-lactam ring is confirmed by infrared analysis.
Preparation 24.Preparali0n 0 o-(a-aminophenylacetamido) -zhi0penicillanic acid 6 (a carbobenzyloxyaminophenylacetamido)penicillanic acid which is obtained by the reaction of equivalent quantities of 6- a-aminophenylacetamido penicillanic acid and benzyl chlorocarbonate in aqueous sodium hy-

Claims (1)

1. A COMPOUND OF THE FORMULA
US353577A 1964-03-20 1964-03-20 Penicillin aldehydes Expired - Lifetime US3316273A (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
BE661405D BE661405A (en) 1964-03-20
US353577A US3316273A (en) 1964-03-20 1964-03-20 Penicillin aldehydes
DE19651545581 DE1545581A1 (en) 1964-03-20 1965-03-17 New penicillin aldehydes and processes for their production
CH385065A CH527846A (en) 1964-03-20 1965-03-19 Penicillanylaldehydes
FR10033A FR1479231A (en) 1964-03-20 1965-03-19 Substituted penicillin aldehydes and process for their preparation
GB11824/65A GB1097338A (en) 1964-03-20 1965-03-19 Penicillanyl aldehydes
NL6503503A NL6503503A (en) 1964-03-20 1965-03-19
AT261265A AT267055B (en) 1964-03-20 1965-03-22 Process for the production of new penicillin aldehydes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US353577A US3316273A (en) 1964-03-20 1964-03-20 Penicillin aldehydes

Publications (1)

Publication Number Publication Date
US3316273A true US3316273A (en) 1967-04-25

Family

ID=23389732

Family Applications (1)

Application Number Title Priority Date Filing Date
US353577A Expired - Lifetime US3316273A (en) 1964-03-20 1964-03-20 Penicillin aldehydes

Country Status (7)

Country Link
US (1) US3316273A (en)
AT (1) AT267055B (en)
BE (1) BE661405A (en)
CH (1) CH527846A (en)
DE (1) DE1545581A1 (en)
GB (1) GB1097338A (en)
NL (1) NL6503503A (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3351587A (en) * 1966-11-14 1967-11-07 American Home Prod Dialkyl-oxo-diazaspiro penicillins
US3351586A (en) * 1965-08-13 1967-11-07 American Home Prod Poly-6(amino-acyl-amino) penicillanic acids and method of preparation
US4115385A (en) * 1975-03-24 1978-09-19 Pfizer Inc. Antibacterial 3-(5-tetrazolyl) penam compounds
US4179511A (en) * 1973-10-17 1979-12-18 Pfizer Inc. Antibacterial 3-(5-tetrazolyl) penam compounds

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2717893A (en) * 1953-09-23 1955-09-13 Bristol Lab Inc Purification of streptomycin
US2739981A (en) * 1952-08-26 1956-03-27 American Home Prod Diamines and salts thereof
US2751378A (en) * 1953-03-11 1956-06-19 Bristol Lab Inc Benzylpenicillin thio acid
US2767168A (en) * 1953-02-20 1956-10-16 Bristol Lab Inc Purification of streptomycin

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2739981A (en) * 1952-08-26 1956-03-27 American Home Prod Diamines and salts thereof
US2767168A (en) * 1953-02-20 1956-10-16 Bristol Lab Inc Purification of streptomycin
US2751378A (en) * 1953-03-11 1956-06-19 Bristol Lab Inc Benzylpenicillin thio acid
US2717893A (en) * 1953-09-23 1955-09-13 Bristol Lab Inc Purification of streptomycin

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3351586A (en) * 1965-08-13 1967-11-07 American Home Prod Poly-6(amino-acyl-amino) penicillanic acids and method of preparation
US3351587A (en) * 1966-11-14 1967-11-07 American Home Prod Dialkyl-oxo-diazaspiro penicillins
US4179511A (en) * 1973-10-17 1979-12-18 Pfizer Inc. Antibacterial 3-(5-tetrazolyl) penam compounds
US4115385A (en) * 1975-03-24 1978-09-19 Pfizer Inc. Antibacterial 3-(5-tetrazolyl) penam compounds
US4143039A (en) * 1975-03-24 1979-03-06 Pfizer Inc. Antibacterial 3-(5-tetrazolyl)penam compounds

Also Published As

Publication number Publication date
NL6503503A (en) 1965-09-21
AT267055B (en) 1968-12-10
GB1097338A (en) 1968-01-03
BE661405A (en)
CH527846A (en) 1972-09-15
DE1545581A1 (en) 1969-07-31

Similar Documents

Publication Publication Date Title
US4504658A (en) Epimerization of malonic acid esters
US3939150A (en) Penicillin derivatives
PL96668B1 (en) METHOD OF PREPARATION OF 6- / D / - / ALPHA-AMINOPHENYLACETAMIDO / -PENICYLATIC ESTER
NO140931B (en) ANALOGICAL PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY EFFECTIVE ESTERS OF ALFA-AMINOBENZYLPENICILLINES
EP0002765A1 (en) Cephalosporin derivatives, process for their preparation and pharmaceutical compositions and their preparation
Doyle et al. 272. Derivatives of 6-aminopenicillanic acid. Part I. α-Aminobenzylpenicillin and some related compounds
CH632513A5 (en) METHOD FOR PRODUCING UNSATURATED DERIVATIVES OF 7-ACYLAMIDO-3-CEPHEM-4-CARBONIC ACID.
US3652546A (en) Esters of 6-aminopenicillanic acid
US3951955A (en) Penicillins substituted with heterocyclic groups
US3316273A (en) Penicillin aldehydes
US4122086A (en) Isopenicillins
US4145418A (en) Thienopyridine substituted cephalosporins
US4193918A (en) Process for the preparation of hydroxy-alpha-amino-benzyl penicillin
CH636100A5 (en) DIHYDROTRIAZINYLTHIOOXACEPHALOSPORINE.
DE2056983B2 (en) S-heterocyclylthiomethyl-cephalosporins, processes for their preparation and pharmaceutical preparations containing them
SU886747A3 (en) Method of preparing cephalosporin compounds
US3406185A (en) Antibiotics
DE2600842C2 (en)
DE3901405A1 (en) CEPHALOSPORINE DERIVATIVES AND METHOD FOR THEIR PRODUCTION
US4959495A (en) Process for the preparation of intermediates used to produce aminothiazoloximino cephalosporins
US3492297A (en) Guanidino cephalosporins
CA1175807A (en) 2.beta.-CHLOROMETHYL-2.alpha.-METHYLPENAM-3.alpha.-CARBOXYLIC ACID SULFONE
CH625805A5 (en)
US3296250A (en) 4-thiazolylmethylpenicillins
US3781277A (en) Dioxo-thiazolidinyl derivatives of 6-aminopenicillanic acid