GB2161803A - Intermediates for beta -lactam antibiotics - Google Patents

Intermediates for beta -lactam antibiotics Download PDF

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GB2161803A
GB2161803A GB08514519A GB8514519A GB2161803A GB 2161803 A GB2161803 A GB 2161803A GB 08514519 A GB08514519 A GB 08514519A GB 8514519 A GB8514519 A GB 8514519A GB 2161803 A GB2161803 A GB 2161803A
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alpha
amino
acid
ethyl
benzyl ester
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GB2161803B (en
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Peter Henry Milner
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Beecham Group PLC
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Beecham Group PLC
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/06—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members
    • C07D241/08—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members with oxygen atoms directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
    • C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
    • C07D311/12—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 3 and unsubstituted in position 7
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D463/00—Heterocyclic compounds containing 1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbacephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D463/10—Heterocyclic compounds containing 1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbacephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2
    • C07D463/14—Heterocyclic compounds containing 1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. carbacephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring with a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2 with hetero atoms directly attached in position 7
    • C07D463/16—Nitrogen atoms
    • C07D463/18—Nitrogen atoms further acylated by radicals derived from carboxylic acids or by nitrogen or sulfur analogues thereof
    • C07D463/20—Nitrogen atoms further acylated by radicals derived from carboxylic acids or by nitrogen or sulfur analogues thereof with the acylating radicals further substituted by hetero atoms or by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D505/00—Heterocyclic compounds containing 5-oxa-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. oxacephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring

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

Abstract

Novel beta -lactam penicillin or cephalosporin compounds having an alpha -amino or alpha -methylthio group are disclosed which are intermediates in the production of beta -lactam antibiotics having an alpha -formamido substituent on the carbon ring adjacent to the carbonyl group of the beta -lactam ring. These antibiotics are disclosed and claimed in UK Patent Application 8221059 (S.N. 2107307) from which this application is divided.

Description

SPECIFICATION Novel compounds This invention relates to novel p-lactam derivatives which are important intermediates in the production of a class of p-lactam antibiotics which are of value in the treatment of infections in animals especially mammals including man, by a wide range of organisms, particularly Gram. negative organisms.
UK Patent application 8221059, from which this application is divided, provides a class of ss-lactam antibiotic having an a-formamido-substituent on the carbon atom adjacent to the carbonyl group of the t3- lactam ring.
The present invention is based on the discovery of novel intermediates for use in the production of oformimado substituted antibiotics.
According to the present invention there is provided a compound selected from the following; 6a-amino-6p-phenoxyacetamido-penicillan acid, the benzyl ester thereof, 6α-amino-6ss-[2-(p-nitrobenzyloxycarbonyl-2-phenylacetamido]penidillanic-acid, the benzyl ester thereof, 6a-amino-6-2-(p-nitrobenzyloxycarbonyl)-2-(3-thienyl)acetamido penicillanic acid, the benzyl ester thereof, 6α-amino-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6α;-amino-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl) acetamido] penicillanic acid, the benzyl ester thereof, 6e-a mino-6ss-[L-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl) acetamido] penicillanic acid, the benzyl ester thereof, 6α-amino-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6α-amino-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-(4-benzyloxycarbonyloxyphenyl) acetamido] penicillanic acid, the benzyl ester thereof, 6α-amino-6ss-[D-2[(4-benzyloxycarbonyloxy)phenyl]-2-(2,2,2-trichloroethoxycarbonylamino)-acetamido] penicillanic acid, the benzyl ester thereof, 6α;-amino-6ss-[DL-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(thien-2-yl)-acetamido]penicillanic acid, the benzyl ester thereof, 6α-amino-6ss-[DL-2-(4-nitrobenxyloxy carbonylamino)-2-(thien-2-yl)acetamido] penicillanic acid, the benzyl ester thereof, 6a-amino-6-[(2R,3S)-3-benxyloxy-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl)carbonylamino]-butyramido] penicillanic acid, the benzyl ester thereof, 6eC-amino-6ss(2t2t2-trichloroethoxycarbonylamino)-penicillanic acid, the benzyl ester thereof, 7cL-amino-7ss-(trichloroethoxycarbonylamino)cephalosporanic acid, the t-butyl ester thereof, 7a-amino-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl) carbonylamino]-2-phenylacetamido]cephalosporan jc acid, the t-butyl ester thereof, 7α-amino-7ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino] acetamido]cephalosporanic acid, the t-butyl ester thereof, 6α-amino-6ss-phenoxyacetamido-penam-3-carboxylic acid, the benzyl ester thereof, 7α-amino-7ss-[D-2-[(4-ethyl-2,3-dioxopi perazin-1 -yl) carbonyla mino]-2-phenylacetamido]-3-methyl-1 -oxadethia-ceph-3-em-4-carboxylic acid, the t-butyl ester thereof, 7a-ami no-7-trich loroethoxycarbonylamino-3-methyl-1 -oxadethia-ceph-3-em-4-carboxylic acid, the t-butyl ester thereof, 6α-amino-6ss(R-2phenyl-2-sulphoacetamido penicillinic acid, triethylammonium salt, the benzyl ester thereof.
Compounds having the general formula (I)
wherein R is an acyl group, R3 and R4 are the same or different and represent C1.14 alkyl or hydrogen, R2 is hydrogen or a carboxylic ester forming group and Y is
where 0 is a methyl or acetoxy methyl group are disclosed in US patent 3,962,214 and further 7a-aminocephalosporin compounds are disclosed in UK patent 1,348,984. However the novel intermediates of the present invention are not disclosed in either patent specification.
The compounds of the present invention may be prepared by the processes disclosed in UK Patent No.
1348984 and US Patent No. 3,962,214.
A preferred process for the preparation of compounds having the partial structure (II)
wherein R is an acyl group, including the 6a-amino-and 7a-amino-intermediates of the present invention is by reaction of the corresponding compound of partial structure (III)
wherein Ras is C16 alkyl, alkyl, or benzyl with anhydrous ammonia, an ammonium salt or an amine of the formula (IV) R20-NH2 (Iv) wherein R20 is a removable protecting group such as benzyl; in the presence of a metal ion such as mercury, silver, thallium, lead or copper and thereafter if necessary removing any protecting group to form the fia-amino-or 7a-amino compound.
Suitable examples of the alkyl group for Ris include C16 alkyl groups such as methyl, ethyl, n-, or isopropyl, and n-, sec-; iso-, or tert-butyl groups.
A preferred alkyl group for Ras is methyl.
Suitable examples of the aryl group R19 include phenyl, optionally substituted with C,.6 alkyl, C,6 alkoxy, halogen, or nitro. Preferred aryl groups for R19 include phenyl, o-, m-or p-methylphenyl, o-, m-or p-nitrophenyl, in particular p-methylphenyl.
Suitable solvents in which the reaction may be performed include for example, diethylether, tetrahydrofuran, dimethylformamide, methanol and hexamethylphosphoramide. The reactions are generally carried out under an inert atmosphere and at moderate to low temperatures i.e. in the range-100 C to 30"C.
The course of the reaction may be followed by convenventional methods such as thin layer chromatography and terminated when an optimum quantity of product is present in the reaction mixture.
The preferred metal ion for use in the above process is the mercuric ion, aptly in the form of mercuric acetate.
Examples of suitable protecting groups for the group R20 include those known in the art as being cleavable to provide the-NH-. Mention may be made of silyl groups such as trimethylsilyl, tertiarybutyl-dimethylsilyl, and tri-isopropylsilyl. A preferred protecting group is (p-methoxymethoxy)phenyl which is removable by cerium ammonium nitrate. Other protecting groups of interest include those cleavable by methanolysis such as-C(CO2R)=O. This moiety may be derived from groups of the type -C(CO2R) =C(CH3)2). Further suitable protecting groups include 4-nitro-benzyl and 2,4-dimethoxybenzyl which is removable with potassium persulphate.
Certain novel compounds having a 6c/-SR16 or 7a-SR'9 group also form a part of the present invention.
Thus according to an aspect of the invention there is provided a compound selected from the following; 6α-methylthio-6ss-[2-(p-nitrobenzyloxy-carbonyl)-2-phenyl-acetamido]penicillanic acid, the benzyl ester thereof, 6α-methylthio-6ss-[2-(p-nitrobenxyloxy-carbonyl)-2-(3-thienyl)acetamidolpenicillanic acid, the benzyl ester thereof, 6α;-methylthio-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6a methylthio-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(4-benzyloxycarbonyloxy- phenyl)-acetamido]penicillanic acid, the benzyl ester thereof, 6a-methylthio-6p-[L-24[(ethyl-2,3-dioxopiperazin-1 -yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl) acetamido]penicillanic acid, the benzyl ester thereof, 6α-methylthio-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6α-(methylthio)-6ss-[DL-2-(4-nitrobenzyloxycarbonylamino)-2-(thien-2-yl)acctamido]penicillanic acid, the benzyl ester thereof, 6α;-(methylthio)-6ss-[2R,3S)-3-benzyloxy-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-butyramido]- penicillanic acid, the benzyl ester thereof 6Ot-(methylthio)-6ss-(2,2,2-trichloroethoxycarbonylamino)-penicillanic acid, the benzyl ester thereof, 7a-methylthio-7p-(trichlornethoxycarbonylamino)cephalosporanic acid, the t-butyl ester thereof, 7α;-methylthio-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)-carbonylamino]-2-phenylacetam- ido]cephalosporanic acid, the t-butyl ester thereof, 7a-methylthio-7ss-[D-2-[4-benzyloxycarbonyloxy) phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl)-carbonylamino)-acetamido]cephalosporanic acid, the t-butyl ester thereof, 6cs-(methylthio)-6ss-amino-penam-3-carboxylic acid, the benzyl ester thereof, 6a-(methylthio)-Sp-phenoxyacetamido)-penam-3-carboxylic acid the benzyl ester thereof, 7ss-(4-nitrobenzylideneamino)-7Oz-(methylthio)-3-methyl-1 -oxadethia-ceph-3-em-4-carboxylic acid, the benzyl ester thereof, 7ss-amino-7cY-(methylthio)-3-methyl-1-oxadethia-ceph-3-em-4 carboxylic acid, toluene p-sulphonic acid salt, the benzyl ester thereof 6α-methylthio-6ss-[D-2-[4-(benxyloxycarbonyloxy)phenyl]-2-(2,2,2-trich loroethoxyca rbo nylamino)acetamido] penicillanic acid, the benzyl ester thereof 6α-methylthio-6ss-[DL-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)-carbonylamino]-2-(thien-2-yl)acetamido] penicillanic acid, the benzyl ester thereof, 7ss-[D-2-[(4-ethyl,2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido-7a-methylthio-3-methyl-1 - oxadethiaceph-3-em-carboxylic acid, the t-butyl ester thereof, 7α-methylthio-7ss-trichloroethoxycarbonylamino-3-methyl-1-oxodethia-ceph-3-em-4 carboxylic acid, the t-butyl ester thereof, 6a-methylthio-6-(R-2-phenyl-2-sulphoacetamido) penicillanic acid, triethylammonium salt, the benzyl ester thereof.
Alternatively compounds of partial structure (II), including the 6a-amino-and 7a-amino-compounds of the present invention may be prepared by the reaction of a compound of partial structure (V)
wherein R and R16 are as hereinbefore defined, Hal is chloro or bromo, with anhydrous-ammonia, an ammonium salt or an amine of formula (lV) as hereinbefore defined and thereafter if necessary removing any protecting group to form the compound of partial structure (II). Compounds of partial structure (V) may be prepared by reaction of the corresponding compound of partial structure (III) with a halogenating agent such as chlorine or bromine in an inert solvent, for example dichloromethane, at a depressed temperature such as-80 C to-30 C.
A further method of preparation of compounds of partial structure (II) including the 6ct-amino-and 7p- amino-compounds of the present invention is by the reaction of the corresponding compound having the partial structure (Vl)
wherein R and R16 are as hereinbefore defined; with anhydrous ammonia, an ammonium salt or an amine of the formula (XII) as hereinbefore defined and thereafter if necessary removing any protecting group to form the compound of the invention.
Suitably such a reaction is performed at a non-extreme temperature for example 0"C-600C normally 109 40 C and preferably ambient. The reaction is conveniently performed in an aprotic solvent such as tetrahydrofuran or dioxan.
It will be appreciated that the processes for the preparation of compounds of partial structure (II) proceed via an imine intermediate; other processes proceeding via such an intermediate are also included herein.
The compounds of partial structure (Vl) may be prepared by the oxidation of a corresponding compound having the partial structure (III). Such oxidation may conveniently be performed in conventional manner, for example using a per-acid such as peracetic acid or m-chloroperbenzoic acid, suitably at an ambient or depressed temperature. Suitable solvents for such a sulphoxidation include ethylacetate, chloroform, dichlormethane, dioxan and tetrahydrofuran.
It will be appreciated that the oxidation of compounds of partial structure (III) having a 6α-SR19 group or a 7cz-SR19 group on a ss-lactam nucleus containing a sulphur atom such as a pencillin or acephalosporin may also oxidise this additional sulphur atom and accordingly it may be necessary to reduce the thus formed suiphoxide or sulphone to the corresponding sulphide.
The compounds of partial structure (III) including the novel 6c -SRa9 compounds of the present invention may be prepared by methods known hitherto, for example by acylation of a compound of partial structure (VII)
wherein Ris is as previously defined.
Compounds of partial structure (VII) may be prepared from a Schiff's base derivative as outlined in Scheme 1.
Scheme 1
The compound of partial structure (VII) is prepared by reacting the amino compound (A) with an aldehyde of formula Ar-CHO wherein Ar is an aryl group to form the Schiff base (B). The Schiff base (B) is reacted with a base to form an anion which is treated with a thiosulphonate of formula: R16S.SO2R1l or a sulphenyl chloride of formula: R'9SCI wherein Rlg is as hereinbefore defined to give the compound of formula (C).Acidic hydrolysis of the Schiff base gives the amino compound of partial structure (VII) Compounds of partial structure (VII) may also be prepared by reacting a thiooxime compound of fomula (VIII):
wherein R19 is as defined hereinbefore above with a tri(alkyl)phosphine or tri(aryl)phosphine, followed by treatment with an acid catalyst such as silica gel. The process is as described in US Patent No. 4,119,778 and i J Amer Chem Soc, 1980, 102, 1690.
Compounds of partial structure (VII) and (VIII) may also be prepared by the process disclosed in US Patent No. 3,962,214 or an appropriate modification thereof.
The following Examples illustrate the preparation and use of the compounds of the present invention.
Example 1 6a-Formamido-6ss-phenoxyacetamido penicillanic acid sodium salt a) Benzyl 60c-amino-6ss-phenoxyacetamido penicillanate Benzyl 6α-methylthi-6ss-phenoxyacetamido penicillanate (389 mg. 0.8 mmol) in DMF (10 ml) at-40 C under nitrogen, was treated with a solution of mecuric acetate (260 mg, 0.8 mmol) in DMF (1 ml), followed immediately by a solution of anhydrous ammonia (15 mg, 0.88 mmol) in DMF (1 ml). The reaction solution was allowed to warm to 0 C over 0.75 hour before being poured into ether and washed with water and brine.The organic solution was dried over magnesium sulphate, filtered and evaporated to leave the essentially pure product as a pale yellow foam (320 mg, 88%), i'max (CH2CI2) 3385, 3310, 1790, 1748, 1690, 1495 cm-l; 8(CDCI3) 1.37 (6H, s, gem dimethyls), 2.78 (2H, br.s, amino protons), 4.51 (3H, s, PhOC,'2-and C-3 proton), 5.19 (2H, s, ester CH2), 5.48 (IH, s, C-S proton), 6.70-7.60 (10H, m, aromatics) and 7.90 (IH, br.s, amido proton).
b) Benzyl 60s-formamido-6ss-phenoxyacetamido penicillanate A solution of benzyl 6a-amino-6(3-phenoxyacetamido penicillanate (320 mg, 0.7 mmol) in anhydrous dichloro-methane (20 ml) at 0 C, was treated sequentially with pyridine (553 mg, 565 AI, 7 mmol) and formic-acetic anhydride (310 mg, 3.5 mmol). The reaction solution was stirred at 0-5"C for 1.5 hour before being washed successively with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution and brine. It was dried over magnesium sulphate, filtered and evaporated to leave the product as a white foam (300 mg).This was purified on silica gel to afford the title compound (240 mg, 71%), i',,, (CH2CI2) 3395, 3310, 1792, 1748, 1700, 1690 (sh.), 1495 cm-1; 8(CDCI3) 1.36 (6H, s, gem dimethyls), 4.51 (3H, s, PhOCH2-and C-3 proton), 5.19 (2H, s, ester CH2), 5.75 (1H, s, C-s proton), 6.70-7.50 (10H, m, aromatics), 8.19 (1H, s, formyl proton), 8.23 (1H, s, 6ss-amido proton) and 8.46 (1H, s, formamido proton).
c) 6a-formamido-6ss-phenoxyacetamido penicillanic acid, sodium salt A solution of benzyl 6a-formamido-6(3-phenoxy-acetamido penicillanate (210 mg, 0.44 mmol) in THF (10 ml) was added to a suspension of 10-% palladium on charcoal (250 mg) in ethanol (10 ml) and water (1 ml) which had been pre-hydrogenated for 0.5 hour. The mixture was hydrogenated for 2.5 hours and then the catalyst was filtered and washed with dilute sodium bicarbonate solution and THF. The organic solvents were evaporated and the aqueous solution washed with ethyl acetate (3 x 30 ml) before being acidified to pH 1.5 with dilute hydrochloric acid. The product was extracted into ethyl acetate (3 x 30 ml) and the combined extracts were washed with brine, dried over magnesium sulphate and evaporated to dryness.The resultant foam (130 mg) was dissolved in acetone and treated with the theoretical amount of 2N sodium ethyl hexanoate in methyl isobutyl ketone (170 iil, 0.34 mmol). Anhydrous ether was then added and the precipitated sodium salt was filtered and washed with ether. The product was dried in vacuo over phosphorous pentoxide to afford 99 mg (55%). Hple showed one peak, Urn,,, (KBr) 1765, 1675, 1600, 1655 cm1. Free acid: 3 [(CD3)2CO] 1.51 (6H, s, gem dimethyls), 4.50 (1H, s, C-3 proton), 4.68 (2H, s, Ph CK2-1), 5.72 (1 H, s, C-S proton), 6.48 (br.s,-CO2H and H2O), 6.80-7.60 (5H, m, aromatics) and 8.25, 8.52 and 8.60 (3H, 3s, amido and formyl protons).
MIC ( > g/ml) P.mirabilis 889 > 100.
Example 2 6(x-Formamido-6ss-(2-carboxy-2-phenylacetamido)penicillanic acid, di-sodium salt a) benzyl 6α-methylthio-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-phenylacetamido]penicillanate A solution of 2-(p-nitrobenzyloxycarbonyl)-2-phenylacetyl chloride (2 mmol) in dichloromethane (10 ml) was added dropwise with stirring to an ice cooled solution of benzyl 6oe-methylthio-6ss-amino penicillanate (0.70 g, 2 mmol) and pyridine (0.24 g, 3 mmol) in dichloromethane (30 ml). The reaction mixture was stirred at 0-5"C for 1 hour, followed for 2 hours at room temperature. It was then washed sequentially with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution and brine, before being dried over magnesium sulphate filtered and evaporated to dryness.The resultant pale yellow foam (1.06 g) was purified on silica gel to afford the pure product as a white foam (0.83 g, 64%), "rn,,,, (CH2Cl2) 3400, 3325, 1788, 1745, 1690, 1530, 1355, 1320 cm-1; 8(CDCI3) 1.30 (6H, br.s, gem dimethyls), 2.11 and 2.20 (3H, 2s,-SCH3 diastereoisomers), 4.42 (1H, s, C-3 proton), 4.85 (1H, s,a-proton), 5.19 (2H, s, benzyl ester CH2), 5.29 (2H, s, PNB ester (CH2), 5.56 (1H, s, C-S proton), 7.10-7.65 (12H, m, aromatics) and 7.90-8.30 (3H, m, aromatics and amido proton).
b) benzyl 6α-amino-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-phenylacetamido]penicillanate A solution of benzyl 6α-methylthio-6ss-[2-p-nitro-benzyloxycarbonyl)-2-phenylacetamido]penicillanate (390 mg, 0.6 mmol) in DMF (10 ml) at-40'C under nitrogen, was treated with mercuric acetate (190 mg, 0.6 mmol) in dry DMF (1 ml) followed immediately by a solution of anhydrous ammonia (11 mg, 0.66 mmol) in DMF (0.5 ml). The mixture was stirred at-40"C to-10 C over 1 hour before being poured into ether and washed with water and brine.It was dried over magnesium sulphate, filtered and evaporated to afford the virtually pure product as a white foram (310 mg, 84%), Vm,,, (CH2CI2) 3395, 3330, 1787, 1742, 1682, 1530, 1357 cm-1; (CDCI2) 1.31 (6H, br.s, gem dimethyls), 2.63 (2H, s, amino protons), 4.42 (1H, s, C-3 proton), 4.73 (1H, s, a-proton), 5.20 (2H, s, benzyl ester CH2), 5.31 (2H, s, PNB ester CH2), 5.41 (1H, s, C-S protoin), 7.20-7.60 (12H, m, aromatics), 7.86 (1H, s, amido proton) and 8.20 (2H, part AA'BB', PNB aromatics).
c) benzyl 6α-formamido-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-phenylacetamido]penicillanate A solution of benzyl 6α-amino-6ss-[2-(p-nitro-benzyloxycarbonyl)-2-phenylacetamido]penicillanate (310 mg, 0.5 mmol) in dry dichloromethane (20 ml) at 0 C, was treated sequentially with pyridine (400 mg, Smmol) and formicacetic anhydride (220 mg, 2.5 mmol). The reaction solution was stirred at 0-5"C for 1 hour, before being washed with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution and brine. It was dried over magnesium sulphate, filtered and evaporated to leave the crude product (250 mg). This was purified on silica gel to afford the title compound as a white foam (220 mg, 69%), vmax (CH2CI2) 3400, 3315, 1795, 1750, 1700, 1690 (sh), 1530, 1355 cm-1; 8(CDCI3) 1.26 (6H, br.s, gem dimethyls), 4.42 and 4.46 (1H, 2s, C-3 proton diastereoisomers), 4.85 (1H, br.s, a-proton), 5.18 (2H, s, benzyl ester CH2), 5.27 (2H, s, PNB ester CH2), 5.70 (1H, s, C-S proton), 7.10-7.60 (12H, m, aromatics) and 7.90-8.70 (5H, m, aromatics, amido and formyl protons); m/e 557, 486, 377, 270, 249, 212, 136, 114, 91.
d) 6α-formamido-6ss-(2-carboxy-2-phenylacetamido) penicillanic acid, di-sodium salt A solution of benzyl 6a-formamido-6(3-[2-(p-nitro- benzyloxycarbonyl)-2-phenylacetamido]penicillanate (200 mg, 0.31 mmol) in THF (10 ml) was added to a suspension of 10% palladium on charcoal (200 mg) in ethanol (10 ml) and water (1 ml) which had been pre-hydrogenated for 1 hour. The mixture was then hydrogenated for 3 hours and then the catalyst was filtered and washed with dilute sodium bicarbonate solution. The filtrate was washed with ethyl acetate (3 x 30 ml) before being acidified to pH 1.5 with 1 N hydrochloric acid.The product was extracted into ethyl acetate (3 x 30 ml) and the combined extracts were washed with brine, dried over magnesium sulphate, filtered and evaporated to dryness to leave the free acid as a colourless oil (110 mgl. This was dissolved in acetone and treated with 2N sodium ethyl hexanoate in methylisobutyl ketone (260 yl, 0.52 mmol) followed by anydrous ether.The resultant precipitate was filtered, washed with ether and dried in vacuo to afford the product as a white solid (103 mg, 72%). Hple showed one peak, vmax (KBr) 1765, 1665, 1600 cm-'. Free acid: 8[(CD3)2CO] 1.10-1.70 (6H, 4s, gem dimethyl diastereoisomers), 4.41 and 4.48 (1H, 2s, C-3 proton diastereoisomers), 4.89 (1H, s, a-pro- ton), 5.66 (1H, s, C-s proton), 7.20-7.80 (m, aromatics,-CO2H and water) and 8.10-9.10 (3H, m, amido and formyl diastereoisomers).
MIC (Lg/ml) P. mirabilis 889 50.
Example 3 6a-Formamido-6ss-[2-carboxy-2-83-thienyl)acetamidol-penicillanic acid, di-sodium salt a) benzyl 6α-methylthio-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-(3-thienyl)acetamido]penicillanate A solution of benzyl 6ss-amino-6α-(methylthio)-pencillanate (1.76 g) and N,N'-dicyclohexylcarbodi-imide (1.13 g) in tetrahydrofuran (10 ml) was stirred and cooled to 0-5"C and treated dropwise with a solution of 2-(p-nitrobenzyloxycarbonyl)-2-(thien-3-yl) acetic acid in tetrahydrofuran (10 ml). The reaction mixture was stirred at 0-5"C for 0.5h, then kept for 18h at 4 C. It was diluted with ethyl acetate and the precipitated N,N'-dicyclohexylurea removed by filtration.The filtrate was evaporated to dryness in vacuo and the resulting, white foam chromatographed on silica gel 60 (230 mesh ASTM) to give the title compound (2.67 g, 81%). vmax (CH2CI2) 3390, 3320, 1785, 1748, 1695, 1525, 1350, 1318 cm-1; 8(CDCI3) 1.31 (6H, s, gem dimethyls), 2.15 and 2.21 (3H, 2s,-SCH3 diastereoisomers) 4.44 (1H, s, C-3 proton), 4.96 and 4.99 (1H, 2s, a-proton diastereoisomers), 5.20 (2H, s, benzyl ester CH2) 5.32 (2H, s, PNB ester CH2), 5.58 (1H, s, C-s proton), 7.10-7.60 (10H, m, aromatics), 7.85 and 7.91 (1H, 2s, amido diastereoisomers) and 8.20 (2H, part AA'BB', PNB aromatics); m/e 655,607, 550, 456, 303, 276, 250, 136, 114, 91.
b) benzyl 6α-amino-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-(3-thienyl)acetamido]penicillanate A solution of benzyl 6ct-methylthio-6ss-[2-(p-nitro-benzyloxycarbonyl)-2-(3- thienyl)acetamido]penicillanate (1.33 g, 2 mmol) in DMF (30 ml) at-40 C under nitrogen, was treated with mercuric acetate (0.64 g, 2 mmol) in dry DMF (3 ml), followed immediately by a solution of anhydrous ammonia (38 mg, 2.2 mmol) in DMF (1 ml). The mixture was stirred at-40"C to-10 C for 1 hour before being poured into ether and washed with water and brine.It was dried over magnesium sulphate, filtered and evaporated to afford the virtually pure product as a foam (1.18 g), 94%) Urn,,,, (CH2CI2) 3380, 3310, 1785, 1745, 1690, 1525, 1350 cm-'; 8(CDC13) 1.33 (6H, s, gem dimethyls), 2.79 (2H, br.s, amino protons), 4.46 (1H, s, C-3 proton), 4.93 (1H, s, a-proton), 5.20 (2H, s, benzyl ester CH2), 5.31 (2H, s, PNB ester CH2), 5.43 (1H, s, C-s proton), 7.10-7.70 (10H, m, aromatics), 7.87 (1H, s, amido proton) and 8:21 (2H, part AA'BB', PNB aromatics).
c) benzyl 6α-formamido-6ss-[2-(p-nitrobenzyloxycarbonyl)-2-(3-thienyl)acetamido]penicillanate A solution of 6a-amino-6(3-[2-(p-nitrobenzyloxycarbonyl)-2-(3-thienyl) acetarhido]penicillanate (1.15 g, 1.8 mmol) in dry dichloromethane (30 ml) at 0 C was treated sequentially with pyridine (1.42 g, 18 mmol) and formic-acetic anhydride (0.79 g, 9 mmol). The reaction solution was stirred at 0-5"C for 2 hours being being washed with 0.5N hydrochloric acid, dilute sodium bicarbonate solution and brine. It was dried over magnesium sulphate, filtered and evaporated to leave the crude product (0.97 g).This was purified on silica gel to afford the title compound as a white foam (0.16 g, 52%) Vrn,,,, (CH2Ci2) 3390, 3310, 1790, 1745, 1695, 1525, 1350 cm-'; 8(CDCI3) 1.28 (6H, m, gem dimethyl diastereoisomers), 4.00 and 4.02 (1H, 2s, C-3 proton diastereoisomers), 4.83 and 4.86 (1H, 2s, a-proton diastereoisomers), 5.14 (2H, s, benzyl ester CH2), 5.23 (2H, s, PNB ester CH2), 5.60 (1H, s, C-s protons), 700-750 (10H, m, aromatics), 7.79 and 7.82 (1H, 2s, exchangeable with D2O, 6ss-amido diastereo-isomers) and 7.97-8.25 (4H, m, 1H exchangeable with D2O, PNB aromatics, formamido and formyl proton); mle 499, 471, 469, 456, 441, 303, 276, 250, 240, 194, 153, 136, 114,91.
d) 6α-formamido-6ss-[2-carboxy-2-(3-thienyl)acetamido] penicillanic acid, di-sodium salt A solution of benzyl 6ct-formamido-6ss-[2-(p-nitro-benzyloxycarbonyl)-2-(3- thienyl)acetamido]penicillanate (0.55 g, 0.84 mmol) in THF (15 ml) was added to a suspension of 10% palladium on charcoal (0.6 g) in ethanol (10 ml) and water (1 ml) which had been pre-hydrogenated for 1 hour, before the addition of further catalyst (0.6 g). The hydrogenation was continued for 2 hours, and then the catalyst was filtered and washed with dilute sodium bicarbonate solution. The filtrate was washed with ether, its pH was then lowered to 1 and the product extracted into ethyl acetate. The combined extracts were washed with brine, dried over magnesium sulphate and evaporated to leave the free acid as a white foam (0.25 g).This was dissolved in acetone and treated with 2N sodium ethyl hexanoate in methyl isobutyl ketone (0.59 ml, 1.18 mmol) followed by ether. The precipitate was filtered, washed with ether and dried in vacuo to afford the title penicillin 0.16 g (40%). Hple showed one peak vmax (KBr) 1765, 1665, 1600, 1550 cm-1; (D2O) 1.20-1.60 (6H, m, gem dimethyl diastereoisomers), 4.25 (1H, m, C-3 diastereoisomers), 5.59 (1H, s, C-s proton), 7.05-7.50 (3H, m, aromatics) and 8.09 and 8.12 (1H, 2s, formyl diastereoisomers).
MIC ( > g/ml) P.mirabilis 889 25.
Example 4 6α-Formamido-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]penicillanic acid, sodium salt a) benzyl 6ol-amino-6P-ID-2-14-eth yl-2,3-dioxopip erazin- 7-yllcarb on ylamino]-2-p hen ylacetamidol-penicil- lanate A solution of benzyl 6α-methylthio-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenyl- acetamido]penicillanate (978 mg, 1.5 mmol) in dry DMF (15 ml) at-40'C under nitrogen, was treated with mercuric acetate (480 mg, 1.5 mmol) in DMF (2 ml) followed immediately by a solution of anhydrous ammonia (28 mg, 1.65 mmol) in DMF (1 ml). The mixture was stirred at-40 C to-10 C over 1 hour before being poured into ethyl acetate and washed with water and brine.It was dried over magnesium sulphate, filtered and evaporated to afford the essentially pure product as a pale yellow glass (710 mg, 76%), vmax (CH2C12) 3380, 3280, 1780, 1740, 1715, 1690 cm-1; (CDCI3) 0.80-1.50 (9H, m, gem dimethyls and-CH2CH2) 2.82 (2H, br.s, amino protons), 3.20-3.80 (4H, m, piperazine CH2 and CH2CH3), 3.85-4.25 (2H, m, piperazine CH2), 4.34 (1H, s, C-3 proton), 5.18 (2H, s, ester CU2), 5.42 (1H, s, C-s proton), 5.58 (1H, d, J=7Hz, a-proton), 7.20-7.60 (11H, m, aromatics and NH) and 10.05 (1H, d, J=7Hz, amido proton).
b) benzyl 6α-formamido-6ss-[D-2-([(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetamido] penicillanate A solution of benzyl 6e-amino-6ss-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1 -yl)carbonylaminoj-2-phenylace- tamido] penicillanate (0.50 g, 0.8 mmol) in anhydrous dichloromethane (30 ml) at OOC, was treated sequentially with pyridine (0.63 g, 8 mmol) and formic-acetic anhydride (0.35 g, 4 mmol). The reaction solution was allowed to warm to room temperature over 3 hours, before being washed with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution and brine.It was dried over magnesium sulphate, filtered and evaporated to leave the crude product (0.45 g). This was purified on silica gel to afford the title compound 0.37 g (71%), p,,,,, (KBr) 3400, 3300, 1785, 1740, 1710, 1680, 1500 cm-1; ô[(CD3)2SO] 0.07-1.20 (9H, m, gem dimethyls and-CH2CH3), 3.20-3.65 (4H, m, piperazin CH2 and CH2CH3), 3.75-4.00 (2H, m, piperazine CH2), 4.40 (1H, s, C-3 proton), 5.14 (2H, s, ester CH2), 5.45 (1H, s, C-s proton), 5.63 (1H, d, J=7Hz, a-pro- ton), 7.20-7.60 (10H, m, aromatics), 8.03 (1H, s, formyl proton), 9.09 (1H, s, exchangeable in D2O, 6ss- amido proton) and 9.96 (2H, m, exchangeable in D2O, -amido and formamido protons).
c) 6α-formamido-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetam- ina]peniclllanic acid, sodium salt A solution of benzyl 6α-formamido-6ss-[D-2-(4-ethyl-2,3-dioxopiperazin-1yl)carbonylamino]-2-plenyl-ace- tamido]penicillanate (200 mg, 0.31 mmol) in THF (15 ml) was added to a suspension of 10% palladium on charcoal (200 mg) in ethanol (10 ml) and water (1 ml) which had been pre-hydrogenated for 15 minutes.
The mixture was then hydrogenated for 2.5 hours, and then the catalyst was filtered and washed with dilute sodium bicarbonate solution. The filtrate was then washed with ethyl acetate, saturated with sodium chloride and acidified to pH 1.5, before extraction of the product into THF/ethyl acetate (50:50).The combined extracts were washed with brine, dried over magnesium sulphate and evaporated to leave the free acid (0.12 g), ô[(CD3)2CO)] 0.90-1.70 (9H, m, gem dimethyls and-CH2CH3), 3.20-4.20 (6H, m, piperazine CU2,s and-CH2CH3), 4.40 (1H, s, C-3 proton), 5.70 (1H, s, C-s proton), 5.82 (1H, d, J=7Hz, a-proton), 7.307.90 (5H, m, aromatics), 8.30 (1H, s, formyl proton), 8.57 (1H, s, 6ss-amido proton), 9.06 (1H, s, formamido proton). This was dissolved in THF and the sodium salt was formed by addition of 2N sodium ethyl hexanoate in methyl isobutyl ketone followed by ether. The product was filtered, washed with ether and dried in vacuo to afford the title penicillin (130 mg 72%). Hple showed one peak, vmax (KBr) 1765, 1710, 1675, 1600, 1515cm-1; (D2O) 0.90-1.50 (9H, m, gem dimethyls and-CH2CH3), 3.10-3.95 (6H, m, piperazin CH2,s and-CH2CH3), 4.02 (1H, s, C-3 proton), 5.37 (1H,'s, C-s proton), 5.50 (1H, s, a-proton), 7.40 (5H, s, aromatics) and 8.07 (1H, s, formyl proton).
MIC (g/ml) P. mirabilis 889 0.2.
Example 5 6α-Formamdo-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxyphenyl) acetam idoipenicillanic acid, sodium salt a) benzyl 6α-methylthio-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-bezyloxycarbony- loxyphenyl)acetamido]penicillanate A solution of D-2-[(4-ethyl-2,3-dioxopiperazin-:1 -yl )carbonylam in ol-2-(4-benzyloxycarbonyloxyphenyl)- acetylchloride (12.2 mmol) in anhydrous dichloromethane (60 ml) was added dropwise to an ice-cooled solution of benzyl 6x-methylthio-6ss-amino penicillanate (13.42 mmol, 1.1 equivalents) and pyridine (1.45 g, 18.3 mmol) in dichloromethane (100 ml). The reaction solution was stirred at 0 C for 0.5 hours followed by 1 hour at room temperature.It was then evaporated to dryness, dissolved in ethyl acetate, and washed with dilute hydrochloric acid, dilute sodium bicarbonate and brine. It was then dried, evapo-rated and chromatographed on silica gel to afford the title compound in 19% yield, together with a 15% yield of the L-isomer.
D-isom,er vmax (CH2CI2) 3250, 1780, 1765, 1750, 1718, 1715, 1680 (sh), 1500, 1220 cm-1; 8[(CD3)2CO] 1.07 and 1.21 (6H, 2s, gem dimethyls), 1.16 (3H, t, J7Hz,-CH2CH3), 2.28 (3H, s,-SCH3), 3.47 (2H, q, Xi7Hz,-CH2CH3), 3.68 (2H, m, piperazin-CH2), 4.01 (2H, m, piperazin-CH2), 4.37 (1H, s, C-3 proton), 5.19 (2H, s, ester-CH2), 5.25 (2H, s, carbonate-CH2), 5.42 (1H, s, C-S proton), 5.71 (1H, d, J7Hz, collapses to singlet on D2O exchange, a-proton), 7.10-7.70 (14H, m, aromatics), 8.77 (1H, s, exchangeable with D2O, 6ss-amido proton), and 10.01 (1H, d, J7Hz, exchangeable with D20, a-amido proton).
L-isomer v,,,,,, (CH2Cl2) 3280, 1750, 1765, 1755, 1720, 1690, 1500, 1215 cm-l; 8[(CD3)2CO] 1.16 (3H, t, J7Hz, CH2CH3), 1.35 and 1.51 (6H, 2s, gem dimethyls), 1.95 (3H, s,-SCH3), 3.47 (2H, q, J7Hz,-CH2CH3), 3.68 (2H, m, piperazin-CH2), 4.01 (2H,m, piperazin-CH2), 4.45 (1H, s, C-3 proton), 5.22 (2H, s, ester-CH2), 5.27 (2H, s, carbonate-CH2), 5.46 (1H, s, C-s proton), 5.72 (1H, d, J7Hz, collapses to singlet on D2O exchange, a-proton), 7.10-7.70 (14H, m, aromatics), 8.82 (1H, s, exchangeable with D2O, 6ss-amido proton) and 10.02 (1H, d, J7Hz, exchangeable with D2O, amido proton).
b) benzyl 6α-amino-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbonyloxy- phenylJacetamido]penicillanate A solution of benzyl 6α-methylthio-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl)carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl)acetamido]penicillanate (1.75 g, 2.18 mmol) in dry DMF (30 ml) at-500C under nitrogen, was treated with mercuric acetate (0.70 g, 2.18 mmol) in DMF (4 ml) followed immediately by a solution of anhydrous ammonia (41 mg, 2.4 mmol) in DMF (2 ml). The mixture was stirred at-50 C to 30"C for 1 hour before being poured into ethyl acetate and washed with water brine.It was dried over magnesium sulphate, filtered and evaported to afford the essentially pure product (1.56 g, 93%), vmax (CH2CI2) 3280, 1780 (sh), 1765, 1750, 1720, 1695, 1680 (sh), 1500, 1220 cm-1; #[(CD3)2CO] 0.80-1.50 (9H, m, gem dimethyls and-CH2CH3), 3.02 (2H, br.s,-NH2), 3.25-3.80 (4H, m, piperazin-CH2 and-CH2CH3), 3.80-4.20 (2H, m, piperazin-CH2), 4.39 (1H, s, C-3 proton), 5.19 (2H, s, ester-CH2), 5.28 (2H, s, carbonate-CH2), 5.43 (1H, s, C-S proton), 5.75 (1H, d, J7Hz, a-proton), 7.10-7.90 (14H, m, aromatics), 8.87 (1H, s, 6ss-amido proton) and 10.15 (1H, d, J7Hz, a-amido proton).
c) benzyl 6α-formamido-6ss-[D-2-(4-ethyl-2,3-dioxopiperazin-1-yl)canbonylamino]-2-(4-benzyloxycarbony- loxyphenyl)acetamido]penicillanate A solution of benzyl 6α-amino-6ss-[D-2-[(4-ethyl-2, 3-dioxopiperazine-l-yl )carbonylamino]-2-4-benzyloxy- carbonyloxyphenyl)acetamido[penicillanate (1.56 g, 2.0 mmol) in anhydrous dichloromethane (60 ml) at 0 C, was treated with pyridine (1.55 g, 20 mmol) and formic-acetic anhydride (0.88 g, 10 mmol). The reaction solution was stirred at 0 C for 0.25 hours followed by 1 hour at room temperature. It was then washed with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution and brine, before being dried over magnesium sulphate, filtered and evaporated to dryness.The crude product (1.39 g) was purified by chromatography to afford the title compound (0.95 g, 59%), vmax (CH2CI2) 3275, 1790, 1770, 1750, 1725, 1715, 1695, 1682 (sh), 1500, 1210 cm-1; 8[(CD3)2CO] 0.97 and 1.18 (6H, 2s, gem dimethyls), 1.17 (3H, t, J7Hz,-CH2CH3), 3.48 (2H, q, J7Hz,-CH2CH2), 3.65 (2H, m, piperazin-CH2), 4.00 (2H, m, piperazine-CH3), 4.39 (1H, d, C-3 proton), 5.18 (2H, s, ester-CH2), 5.26 (2H, s, carbonate-CH2), 5.58 (1H, s, C-S proton), 5.73 (1H, d, J7Hz, collapses to singlet on D2O exchange, a-proton), 7.10-7.70 (14H, m, aromatics), 8.16 (1H, s, NHCHO], 8.23, (1H, s, exchangeable with D2O,-NHCHO), 8.88 (1H, s, exchangeable with D2O, 6ss-amido proton), and 10.05 (1H, s, J7Hz, exchangeable with D2O,-amido proton).
d)α-formamido-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxyphenyl)acetamido] penicillanic acid, sodium salt A solution of benzyl 6α-formamido-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzy- loxycarbonyloxyphenyl)acetamido]penicillanate (0.50 g, 0.625 mmol in THF (15 ml) was added to a suspensionof 10% palladium on charcoal (0.50 g) in ethanol (15 ml) and water (1 ml) which had been prehydrogenated for 1/2 hour. The mixture was then hydrogenated for 45 minutes, and the catalyst was filtered and washed with dilute sodium bicarbonate solution. The filtrate was then washed with ethyl acetate, saturated with sodium chloride and acidified to pH 1.5, before extraction of the product into THF/ ethyl acetate (50:50).The combined extracts were washed with brine, dried over magnesium sulphate and evaporated to leave the free acid as a white solid (0.25 g). This was suspended in water and the pH carefully adjusted from 2.0 to 7.0 by addition of dilute sodium bicarbonate solution. The resulting solution was filtered and freeze dried to afford the title penicillin 256 mg (69%). Hplc showed one major peak, vmax (KBr) 1770, 1710, 1685, 1670, 1610, 1510 cm (D2O) 0.95 and 1.33 (6H, 2s, gem dimethyls), 1.21 (3H, t, J7Hz,-CH2CH3), 3.50 (2H, q, J7Hz,-CH2CH3), 3.65 (2H, m, piperazin-CH2), 3.98 (2H,m, piperazin-CH2), 4.16 (1H, s, C-3 proton), 5.37 (1H, s, C-s proton), 5.59 (1H, s, a-proton), 6.86 and 7.35 (4H, AA'BB', J9Hz, aromatics) and 8.00 (1H, s,-NHCHO).
MIC ( g/ml) P.mirabilis 889 0.1.
Example 6 6α-Formamido-6ss-[L-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxy- phenyl)acetamidolpenicillanic acid, sodium salt a) benzyl 6α-amino-6ss-[L-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbonyloxy- phenyl)acetamindo]penicillanate This compound was prepared in 84% yield by reaction of benzyl 6α-methylthio-6ss-[L-2-[(4-ethyl-2,3- dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl)acetamido]penicillanate with ammonia in the presence of mercuric acetate as described for the D-isomer. vmax (CH2CI2) 3280, 1780, 1765, 1750, 1715, 1690, 1680 (sh), 1495, 1215 cm-1;; #[CDCl3] 0.95- 1.40 (9H, m, gem dimethyls and-CH2CH3), 2.61 (2H, s,-NH2), 3.10-3.70 (4H, m, piperazine-CH2 and-CH2CH3), 3.75-4.15 (2H, m, piperazine-CH2), 4.31 (1H, s, C-3 proton), 5.11 (2H, s, ester-CU'2), 5.19 (2H; s, carbonate-CH2), 5.33 (1H, s, C-s proton), 5.45 (1H, d, J6Hz, a-proton), 6.90-7.70 (14H, m, aromatics), 8.22 (1H, s, 6ss-amido proton) and 9.81 (1H, d, J6Hz, e-amido proton).
b) benzyl 6α-formamido-6ss-[L-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbony- loxyphenyl)acetamidolpenicillanate This compound was prepared in 61% yield by reaction of benzyl 6α-amino-6ss-[L-2-[(4-ethyl-2,3-dioxopi- perazin-1 -yl)carbonylamino]-2-(4-benzyloxycarbonyloxy-phenyl)acetamido]penicillanate with formic-acetic anhydride in the presence of pyridine, as described forthe D-isomer. vmax (CH2CI2) 3280, 1790, 1765, 1750, 1715, 1690, 1500, 1480, 1215 cm-1;; # [(CD3)2CO] 1.13 (3H, t, J7Hz,-CH2CH3), 1.31 (6H, s, gem dimethyls), 3.45 (2H, q, J7Hz,-CH2CH3), 3.62 (2H, m, piperazine-CH2), 4.00 (2H, m, piperazine-CH2), 4.46 (1H, s, C-3 proton), 5.19 (2H, s, ester-CH2), 5.26 (2H, s, carbonate-CH2), 5.61 (1H, s, C-s proton), 5.75 (1H, d, J7Hz, collapses to singlet on D2O exchange,-proton), 7.10-7.65 (14H, m, aromatics), 8.08 (1H, s,-NHCHO), 8.31 (1H, s, exchangeable with D2O,-NHCHO), 8.60 (1H, s,. exchangeable with D2O, 6ss-amindo proton), and 9.99 (1H, d, J 7Hz, exchangeable with D2O, a-amido proton).
c) 6a-formamido-6ss-[L-2-[64ethyl-2,3-dioxopiperazin- 1-yl)carbonylamino]-2-(4-h ydroxy phenyl)acetamido]-penicillanic acid, sodium salt This compound was prepared in 44% yield by hydrogenolysis of benzyl 6α-formamido-6ss-[L-2-[(4-ethyl- 2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl)acetamido]penicillanate in the manner described for the D-isomer, vmax (KBr) 1770, 1710, 1675, 1610, 1510 cm-1;; #(D2O) 1.13 (3H, s, gem dimethyl), 1.21 (3H, t, J7Hz,-CH2CH3), 1.43 (3H, s, gem dimethyl), 3.52 (2H, q, J7Hz,-CH2CH3), 3.69 (2H, m, piperazine-CH2), 4.01 (2H, m, piperazine-CH2), 4.22 (1H, s, C-3 proton), 5.49 (1H, s, C-s proton), 5.58 (1H, s, a-proton), 6.93 and 7.36 (4H, AA'BB', J9Hz aromatics) and 8.11 (1H, s,-NHCHO).
MIC ( g/ml) P.mirabilis 889 125.
Example 7 6a-Formamido-6(3-(D-2-amina-2-phen ylacetamida)-peniclllanic acid a0 benzyl 6α-amino-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanate Benzyl 6α-methylthio-6ss-[D-2-(4nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanate (1.8 g, 30 mmol) in DMF (25 ml) at-40 C under nitrogen was treated with a solution of mercuric acetate (0.98 g, 31 mmol) in DMF (3 ml). The reaction mixture was allowed to warm to 0 C over 1 hr., poured into ether, washed with water and brine, dried over magnesium sulphate and evaporated to give the title compound (0.97 g, 57%), b(CDCI3), 0.96, 1.19 (6H, 2s, gem dimethyls), 2.74 (2H, m, NH2), 4.38 (1H, s, C-3 proton), 5.15 (4H, s, ArCU2), 5.41 (1H, s, C-S proton), 5.53 (1H, m, a-proton), 6.89 (1H, m, NH), 7.36 (12H, m, aromatic protons), 7.90-8.40 (3H, m, NH aromatic protons); vmax (CH2CI2) 1610, 1680, 1715, 1745, 1780 cm-1.
b) benzyl 6α-formamido-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanate Benzyl 6α-amino-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanate (0.97 g, 1.65 mmol) in dichloromethane (40 ml) at 0 C was treated with pyridine (1.3 ml, 16.5 mmol), then acetic formic anhydride (0.73 g, 8.2 mmol). The reaction mixture was stirred at 0-5 C for 1 hr., washed with 0.5N hydrochloric acid, dilute sodium bicarbonate solution and brine, dried over magnesium sulphate and evaporated to give the crude product.Purification on silica get gave the title compound (0.60 g, 59%), 8(CDCl2) 0.93, 1.17 (6H, 2s, gem dimethyls), 4.41 (1H, s, C-3 proton), 5.16 (4H, s, ArCH2), 5.64 (2H, m, aand C-s protons), 6.89 (1H, s, NH), 7.39 (12H, m, aromatic protons), 8.14 (3H, m, aromatic protons and CHO), 8.85 (1H, m, NH); vmax (CH2CI2) 1605, 1690, 1740, 1785 cm-1, c) 6a-formamido-6(3-(D-2-amino-2-pheny/acetamido)-peniclllanic acid A solution of benzyl 6α-formamido-6ss-[D-2-(4-nitro benzyloxycarbonylamino)-2-phenylacetam ido]penicillanate (0.60 g, 0.9 mmol) in THF (5 ml), ethanol (10 ml), water (2 ml) was added to 10% palladium on charcoal (0.7 g) in THF (2.5 ml), ethanol (5 ml), water (1 ml) which had been prehydrogenated for 15 minutes.The mixture was hydrogenated for 2 hr., further catalyst (0.5 g) was added and hydrogenation continued for 2 hr. The catalyst was filtered and washed sequentially with THF, ethanol, water. The organic solvents were evaporated from the filtrate and the aqueous solution washed with ethyl acetate and freeze dried to give the title compound (0.33 g, 93%), 3(D2O) 0.96, 1.36 (6H, 2s, gem dimethyls), 4.19 (1H, s, C-3 protons), 4.94 (1H, s, a-proton), 5.63 (1H, s, CHO); p,,,, (KBr) 1600, 1675, 1765 cm-1.
MIC ( > g/ml) P. Mirabilis 889 25.
Example 8 6α-Formamido-6ss-[D-2-amino-2-(4-hydroxyphenyl)acetamido]penicillanic acid a ) Benzyl 6a-amin a-6(3-fD-2- (4-nitrob enzyloxycarb on yla min o)-2-(4-b enzyloxycarb an yloxy- phenyl)acetamido]-penicillanate Benzyl 6α-amino-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-(4-benzyloxycarbonyloxphenyl)acetamido] penicillanate (0.84 g, 1.03 mmol) in DMF (20 ml) was cooled to-40 C, treated with mercuric acetate (0.34 g, 1.03 mmol) in DMF (3 ml) followed by anhydrous ammonia (0.019 g, 1.1 mmol) in DMF (1.1 ml).The reaction mixture was allowed to warm to 0 C over 1.5 hours poured into ether, washed with water, brine, dried over magnesium sulphate and evaporated to give the title compound (0.35 g, 43%), 8(CDCI3) 0.96, 1.19 (6H, 2s, gem dimethyls), 2.57 (1H, m, NH2), 4.34 (1H, s, C-3 proton), 5.16 (5H, mArCH2 and-proton), 5.39 (1H, s, C-S proton), 6.63 (1H, m, NH), 7.37, 7.42 (12H, 2s + m, aromatic protons and NH), 8.08 (2H, m, aromatic protons); p,,,,, (CH2CI2) 1610, 1680, 1770 cm-1.
b) Benzyl Ga-formamido-GP-[D-2- (4-nitrob enzylox ycarb on ylamin o)-2-(4-benzyloxycarb on yloxyph en yll- acetamido] penicillanate A solution of benzyl 6α-formamido-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-(4-benzyloxycarbonyl-ox- yphenyl)acetamido]penicillanate (0.35 g, 0.45 mmol) in dichloromethane (10 ml) was cooled to 0 C, treated with pyridine (0.32 ml, 4.0 mmol) and acetic formic anhydride (0.19 g, 2.2 mmol). The reaction mixture was stirred at 0.5 C for 1 hour, washed with 0.5 N hydrochloric acid, dilute sodium bicarbonate solution, brine, dried over magnesium sulphate and evaporated.The crude product was purified on silica gel to give the title compound (0.18g, 50%), 8(CDCI3) 0.94, 1.15 (6H, 2s, gem dimethyls), 4.34 (1H, s, C-3 proton), 5.10 (2H, s, ArCH2), 5.21 (2H, s, ArCH2), 5.55 (2H, m, C-S and a-protons), 7.31, 7.39 (14H, 2s + m, aromatic protons and NH, 7.99 (3H, aromatic protons and CHO), 8.86 (1H, m, NH); vamx (CHCI2) 1610, 1710, 1750, 1770, 1790 cm-'.
c) 6α-formamido-6ss-[D-2-amino2-(4-hydroxyphenyl)-acetamido]penicillanic acid Benzyl 6α-formamido-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-(4-benzyloxycarbonyloxy- phenyl)acetamido] penicillanate (0.29 g, 0.36 mmol) in THF (2 ml), ethanol (5 ml), water (1 ml) was added to a suspension of 10% palladium on charcoal (0.3 g) in THF (2 ml), ethanol (5 ml), water (2 ml) which had been prehydrogenated for 15 minutes. The mixture was hydrogenated for 4 hr. the catalyst was filtered and washed successfully with THF, ethanol, water.The organic solvents were evaporated from the filtrate and the aqueous residue washed with ethyl acetate and freeze dried to give a quantitative recovery of the title compound. 8(D2O) 0.97, 1.28 (6H, 2s, gem dimethyls), 6.14 (1H, s, C-3 proton), 5.25 (1H, s, a-proton), 5.55 (1H, s, C-S proton), 6.83, 7.27 (4H, AA'BB', J10Hz, aromatic protons), 8.18 (1H, s, CHO); vmax (KBr) 1600, 1670, 1765 cm-1.
MIC (llg/ml) P. Mirabilis 889 50.
Example 9 6ss-[D-2-(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(thien-2-yl)acetamido]-6&alpha;-formamidopenicillanic acid, sodium salt a0 Benzyl 6ss-[DL-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(thien-2-yl)acetamido]-6&alpha;-(methyl- thio)penicillanate A solution of benzyl 6p-amino-6a-(methylthio)penicillanate (1.46 g) and N, N'-dicyclohexylcarbodiimide (0.92 gin dichloromethane (25 ml) was stirred and cooled to 0-5 C and treated, dropwise, with a solution of DL-2-[(4-ethyl-2.3-dioxopiperazin-1-yl)carbonylamino]-2-(thien-2-yl)acetic acid (1.35 g) in acetone (5 ml) and dichloromethane (25 ml) over 1 hr. The reaction mixture was allowed to regain room temperature over 1h and stirred at room temperature for 18h.The precipitated N, N'-dicyclohexylures was removed by filtration and washed with dichloromethane (20 ml). The organic filtrate was concentrated in vacuo and the residue chromatographed on silica gel 60 ( < 230 mesh ASTM) eluting with ethyl acetate to give the title compound (1.291 g); v,,,. (KBr) 3450 br, 3280, 1778, 1740, 1710, 1680, 1510 and 1185 cm-1;; 3 [(CD2)2CO] 1.04-1.60-(9H, m, C(CH3)2 and NCH2CH3), 2.05 and 2.28 (3H, 2s, SCH2),3.46 (2H, q, J 8 Hz, NCH2CH2), 3.68 and 4.02 (4H, 2m, NCH2CH2N), 4.39 and 4.44 (1H, 2s, 3-H), 5.19 and 5.21 (2H, 2s, PhCH2,s), 5.45 (1H, s,5-H), 5.97 (1H, d, J7 Hz, CH), 6.88-7.03 (1H, m, thienyl-H), 7.15-7.50 (7H, m, phenyl and thienyl-H2), 8.81 (1H, d,J7 Hz NH), and 9.79-10.02 (1H, bx, CONH).
b) Benzyl 6&alpha;-amino-6ss-[DL-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbon ylamino]-2-(thien-2- yl)acetamido]penicillanate A solution of benzyl 6ss-[DL-2-(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(thien-2-yl)acetamido]- 6(x-(methylthio) pencillanate (1.3 g) in N, N-dimethylformamide (13 ml) was cooled to-40 C under nitrogen and stirred. Mercuric acetate (0.63 g), followed by a solution of ammonia in N, N-dimethylformamide (2.2 ml of 16 mg!ml), was added and the mixture stirred at-20 to-40 C for 0.5 hr. The reaction mixture was allowed to regain room temperature over 1h and diluted with ethyl acetate (100 ml).The mixutre was filtered and the filtrate washed five times with water, saturated brine, and dried over anhydrous magnesium sulphate. The solution was concentrated in vacuo to yield the title compound (1.16 g); 8[(CD3)2CO] 1.00-1.50 (9H, m, C(CH3)2 and NCH2 CH3), 2.67-3.10 (2H,brs,NH2),3.27-3.82 and 3.87-4.19(6H,m, piperazine CH2,s), 4.36 and 4.41 (1 H,2s, 3-H),5.20 (2H, s, CH2Ph), 5.37 and 5.39 (1 H, 2s, 5-H), 5.91(1 H, d, J 7 Hz, CH), 6.78-7.07 and 7.10-7.45 (8H, m, phenyl and thienyl-H's), 8.50 and 8.60 (1H, 2s, CONH), 9.9C(1H, d, J 7Hz, NHCH).
(c) Benzyl 6P-[DL-2-[(4-ethyl-2,3-dioxopiperazin- l-yl)carbon ylaminoj-2-(thien-2-yl)acetamidoi-6a-formam- idopenclllanate Pyridine (1.5 ml) was added to a solution of benzyl 6(x-amino-6ss-[DL-2-[(4-ethyl-2,3-dioxopiperazin-1- yl)carbonylamino]-2-(thien-2-yl )acetamido]pencillanate (1.16 g) in dichloromethane (20 ml) at 0-50C. The solution was treated with acetic formic anhydride (0.81 9) and the mixture stirred at 0-5 C for 1 h, then allowed to regain room temperature over 0.5 h. It was washed with M. hydrochloric acid, dilute aqueous sodium hydrogen carbonate, water, saturated brine, and dried over anhydrous magnesium sulphate.The solution was concentrated in vacuo and chromatographed on silica gel 60 ( < 230 mesh ASTM), eluting with ethyl acetate, to give the separated diastereoisomers.
The first to elute was the diastereoisomer with the side-chain in the L-configuration (0.44 g); [(x12g + 71.5 (c 1 in CHCI3); , max. (KBr), 3280 br, 1782, 1740, 1710, 1680 br, 1505 br, 1460, 1392, 1368, 1324 br, 1265 br, 1200 sh, and 1187 cm-1; 3 (CDCI3) 1.04-1.36 (9H, m, C(CH3)2 and CH2CH3), 3.31-3.65 (4H, m, NCH2CH2N), 3.90- 4.15 (2H, m, CHCH3), 4.39 (1H, s, 3-H), 5.13 (2H, s, CH2Ph), 5.61 (1H, s, 5-H), 5.78 (1H, d, J 8Hz, NHCH), 6.85-700 and 7.11-7.40 (8H, m, phenyl and thienyl-H's), 7.75 t1H, s, CONH), 8.02 (1H, s, NHCHO), 8.11 (1H, s, CHO), 9.82 (1H, d, J 8 Hz, NHCH). The diastereoisomerwith the D-configuration in the side-chain eluted second 0.33 g); [ot]2g + 79.8 (C 1 in CHCI3); vmax. (KBr) 3400 sh, 3280, 1782, 1740, sh, 1710, 1680 br, 1505, 1460, 1391, 1368, 1330, 1280, 1262, and 1185 cm-1; 3 (CDCI2) 0.98 and 1.10-1.37 (9H, m, C(CH3)2 and CH2CH3), 3.25-3.98(6H, m, NCH2 CH2NCH2), 4.38 (1H,s, 3-H), 5.12 (2H, s, CH2Ph), 5.54 (1H, s, 5-H), 5.82 (1H, d, J 8Hz, NHCH), 6.80-7.00 and 7.10- 7.42 (8H, m, phenyl and thienyl-H's), 8.13 (2H, brs, NHCHO), 8.58 (1H, brs,CONH), 10.00 (1H, d, #8 Hz, NHCH).
(d) 6ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(thien-2-yl)acetamido]-6&alpha;-formamidopeni- cillanic acid, sodium salt Benzyl 6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(thien-2-yl)-acetamido]-6&alpha;-formamido- pen-icillanate (280 mg) in tetrahydrofuran (20 ml) containing water j2 ml) was hydrogenated for 4h over 10% palladium on carbon (280 mg). The catalyst was removed by filtration, washed with tetrahydrofuran and the filtrate and washings combined and concentrated in vacua. The aqueous residue was covered with ethyl acetate and the pH adjusted to 8 with M. sodium hydrogen carbonate solution. The phases were separated, the aqueous phase washed with ethyl acetate and ether and acidified to pH2 with 5M.
hydrochloric acid in the presence of ethyl acetate, The phases were separated, the aqueous phase further extract-ed with ethyl acetate, the extracts combined, washed with water at pH2, saturated brine and dried over anhydrous magnesium sulphate. The solution was evaporated to dryness in vacuo to yield 6p-[D-2-[(4-ethyl-2,3-dioxopiperazin-l- yl)carbonylamino]-2-(thien-2-yl)acetam idoJ-6c-formidopencil lanic acid (70 mg). The acid was suspended in water (30 ml), the pH adjusted to 6.1 by addition of M. aqueous sodium hydrogencarbonate and after 15 min the mixture was filtered and the filtrate freeze-dried to yield the title compound (58 mg); v max.
(KBr) 3740-2600, 1770, 1720, 1680 br, 1610, 1510, 1400, 1370, and 1190cm-1; 8 (D2O) 1.08 and 1.32 (6H, 2s, C(CH3)2), 1.18 (3H, t, J 8 Hz, NCH2CH2), 3.50 (2H, q, J 8Hz, NCH2CH2), 3.69 (2H, brt, NCH2CH2N), 4.00 (2H,brt, NCH2CH2N), , 4.20 (1H, s, 3-H), 5.59 (1H, s, 5-H), 5.78 (1H, s, CH), 7.07, 7.28, and 7.49 (3H, m, thienyl-H's), 8.10 (1H, s, CHO).
MIC (llg/ml) P. Mirabilis 889 0.1.
Example 10 6ss-[D-2-Amino-2-(thien-2-yl)acetamido]-6a-formamidopencillanic acid (a) Benzyl-6&alpha;-(methylthio)-6ss-[DL-2-(4-nitrobenzyloxycarbonylamino)-2-(thien-2- yllacetamido]penicillanate A solution of benzyl 6ss-amino-6&alpha;-methylthiopenicillanate (3.52g)and N,N'-dicyclohexylcarbodiimide(2.26g) in acetone (20ml) was stirred and cooled to 0-5"C and treated dropwise, over 1h, with a solution of DL-2-(4-nitrobenzyloxycarbonylamino)-2-(thien-2-yl)acetic acid (3.269) in acetone (20ml). The mixture was allowed to regain room temperature over 1h, then stirred at room temperature for 18h.The precipitated N,N-dicyclohexylurea was removed by filtration, washed with acetone and dried (1.679). The filtrate and washings were combined and evaporated to dryness in vacuo. Chromatography on silica gel 60 ( < 230 mesh ASTM), eluting with 20% ethyl acetate in cyclohexane gave the title compound (5.1g); vmax (KBr) 3400sh, 3050, 1780, 1745, 1720sh, 1680, 1520, 1348, 1230, 1205, and 1183cm-1; 8(CDCI3) 1.10, 1.25, 1.30 and 1.35 (6H,4s,C(CH2)2), 2.02 and 2.19 (3H,2s,SCH2) 4.34 and 4.40 (1H,2s,3-H), 5.15,5.16 and 5.22 (4H,2s and brs, 2CH2,s), 5.53(1H,s,5-H),5.72-5.95 (1H,m,CH), 6.31(1H,d,J8Hz, NHCH), 6.82-7.60 and 8.058.25(12H,m,C6H4, phenyl and thienyl-H's); (found:M 670.1194, C30H29N4O8S3 requires M 670.1178).
(b) Benzyl 6&alpha;-amino-6ss-[DL-2-(4-nitrobenzyloxycarbonylamino)-2-(thien-2-yl)acetamido]penicillanate Benzyl 6a-(methylthio)-6ss-[DL-2-(nitrobenzylOxyCarbonylamino)-2-(thien-2-yl)acetamidolpenicillanate (5.1g) was dissolved in N,N-dimethylformamide (40ml), cooled to 0-5"C and treated with mercuric acetate (2.4g) then ammonia (126mg) in N,N-dimethylformamide (6ml). It was stirred at 0-5"C for 15 min then diluted with ethyl acetate (150ml), filtered and the phases separated. The ethyl acetate layer was washed five times with water, saturated brine, dried over anhydrous magnesium sulphate and evaporated to dryness in vacuo.Chromatography on silica gel6Q( < 230 mesh ASTM), eluting with 50% ethyl acetate in cyclohexane, gave the title compound (3.3g); ",,(KBr) 3370, 3310, 1772, 1738, 1725sh, 1675br, 1520, 1350, 1264, 1240sh, 1210 and 700cm-1.
(c) Benzyl 6&alpha;-formamido-6ss-[DL-2-(4-nitrobenzyloxycarbonylamino]-2-(thien-2- yl)acetamidoapenicillanate.
Treatment of a solution of benzyl 6&alpha;-amino-6ss-[DL-2-(4-nitrobenzyloxycarbonylamino)-2-(thien-2- yl)acetamido]penicill-anate (3.09) in dichloromethane (25ml) at 0.5-C with pyridine (3.7ml), followed by acetic formic anhydride (1.72ml), was carried out under nitrogen. After 0.5h at 0-53C, the mixture was allowed to regain room temperature over 1h, washed with M. hydrochloric acid, M. aqueous sodium hydrogen carbonate, saturated brine, dried over anhydrous magnesium sulphate, and evaporated to dryness in vacuo. Purififcation and separation of the diastereoisomers was accomplished using a Waters prep 500 high performance liquid chromatograph with a PrepPak-500,silica column as the stationary phase and 50% ethyl acetate in cyclohexane as the eluant.The material with the D-configuration in the side-chain eluted first (1.15g); [ce]20 + 168.5 (C 1 in MeOH): vmax (KBr) 3300br 1780, 1740, 1700, 1680, 1520, 1350, 1247, 1210, 1185 and 700cm-1; 8(CDCI3) 1.07 and 1.26 (6H,2s,C(CH2)2), 4.38 (1H,s,3-H), 5.13 (4H,brs,2CH2,s), 5.60 (1H,s,S-H), 5.68 (1H,d,J8Hz, CH), 6.81-7.49 (10H, m, phenyl, thienyl and 2 C6H4 protons), 7.65(1H,brs,CONH), 7.70-8.00(1H, NHCHO0, 8.01-8.20 (3H,m,CHO and 2 C6H4 protons).The title compound with the L-configuration in the side-chain eluted second (0.92g); [&alpha;]D20 + 112.80 (C 1 in MeOH); p,,,,, (KBr) 3300br,1780, 1735, 1680br, 1605, 1520, 1350, 1250br, 1210,and 700cm-1; 8(CDCI3) 1.15 and 1.21 (6H,2s, C(CH3)2), 4.39 (1H,s,3-H), 5.12(4H,brs,2CH2,s), 5.60 (1H,s,S-H), 5.79(1H,d,J8Hz,CH), 6.30-6.70 (1H,br,NHCH), 6.81-6.96(2H,m, thienyl-H2), 7.08-7.49 (8H,m,thienyl-H, phenyl and 2 C6H4 protons), 7.82 (1H,brs, CONH), 8.00-8.20 (2H,m,CHO and 2 CH4 protons), 8.30-8.80 (1H,br,NHCHO.
(d) 6i8-JD-2-A mino-2-(thien-2-yl)acetamidoj-6a-farmamidop enicillanic acid Benzyl 6a-formamido-6(3-[D-2-(4-nitrobenzyloycarbonylamino)-2-(thien-2-yI)acetamidojpenicillanate (645mg) was dissolved in tetrahydrofuran (10ml) and the solution diluted with water (2ml), before hydrogenation over 10% palladium on carbon (650mg) for 4h. The catalyst was removed by filtration, washed with tetrahydrofuran and water, the filtrate and washings combined, concentrated, washed well with ethyl acetate and ether and freeze dried to yield the title compound (175mg); vmax (KBr) 3740-2200, 1770, 1675br, 1604, 1500, 1390, 1340 and 1250 cm-1.
Example 11 6ss-[(2R, 3S)-3-Benzyloxy-2-[4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]butyramido]-6&alpha;-formamidopeni- cillanic acid, sodium salt (a) 62R, 3S)-3-Benzyloxy-2-[(4-ethyl-2,3-dioxopiperazin- l-yl)carb on ylamin o]b utyric acid O-Benzyl-D-threonine (0.92 g, 4.4 mmole) (prepared by the method of T. Mizoguchi, G. Levin, D.W.
Woolley, and J.M. Stewart, J. Org. Chem., 1968, 33, 903, who described the L-isomer) was suspended in water (20 ml) and the pH adjusted to 9.5, giving a virtually complete solution. This was cooled to OOC and stirred while 4-ethyl-2,3-dioxopiperazine-1-carbonyl chloride (1.00 g, 4.9 m mole) in AR acetone (10 ml) was added dropwise. The pH was kept at 9.5 by the addition of further base while the solution was allowed to regain room temperature. After 2h the solution was acidified to pH 2 and the bulk of the acetone removed by evaporation. The oily product was extracted into ethyl acetate (3 x 50 ml), then the total organic phase was washed once with brine and dried over sodium sulphate.Evaporation gave the title acid (1.36 g, 82%) as a colourless non-crystalline foam which retained solvent traces tanaciously; R0.35 in chloroform: methanol:acetic acid, 17:2:1; 8 (CDCI2) 0.75-1.35 (5H, m, CH3CH and NCH2CH3), 3.1-3.7 (4H, m, 2 NCH2), 3.9-4.25 (2H, m, NCH2), 4.50 (2H, m, CH-CH), 6.25 (1 H, brs, D2O exch, OH), 7.25 (SH,s,aryl), 9.50 (1H, brd),J7 Hz, D2O exch, NH), 9.70 (1H, brs, D2O exch, CO2H).
(b) Benzyl 6ss-[(2R,3S)-3-benzyloxy-2-[(4-ehyl-2,3-dioxopiperazin-1-yl)carbonylamino]butyramido]-6&alpha;- (methylthio)pencillanate The preceding acid benzyl ether (1.29 g, 3.42m mole) was dissolved in dry dichloromethane (7 ml) and added dropwise with stirring at 0 C over 0.5 hr to a solution of benzyl 6ss-amino-6&alpha;-(methyl- thio)penicillanate (1.20 g, 3.4 m mole) and dicyclohexylcarbodi-imide (0.70 g, 3.4m mole) in dichloromethane (5 ml). The mixture was allowed to regain room temperature and stirring was continued overnight.
After 16h the precipated dicyclohexylurea was filtered off and the filtrate evaporated to dryness. The residue was dissolved in ethyl acetate and the filtered solution washed sequentially with water, 0.5M. hydrochloric acid, water, saturated aqueous sodium hydrogen carbonate, water and brine, then dried over sodium sulphate.Evaporation gave crude product which was chromatographed on silica gel (120 g), eluting with 2% methanol in chloroform, to give the title 6ct-(methylthio)penicillin ester as a colourless, crisp foam (0.59g, 24%); R, 0.60 in 10% methanol-chloroform; 8 (CDCI3) 1.17 (3H, t,J7 Hz, NCH2CH2), 1.26 (3H, d, J 7Hz, CH3CH), 1.32 and 1.45 (6H, 2s, (CH2)2C), 2.12 (3H, s, CH2S) , 3.50 (4H, m, 2 NCH2), 3.90-4.30 (3H, m, NCH2+CH(CH3)), 4.42 (1H, s, 3-H), 4.45-4.65 (3H, m, Ph CH2O (ether) + CHNH), 5.15 (2H, s, Ph CH2O), 5.51 (1H, s, 5-H), 7.27 and 7.32 (10H, 2s,phenyls), 7.62 (1H,brs,D2O exch, 6-NH), 9.50(1H,brd, J 7Hz, D2O exch, CH-NH).
(c) Benzyl 6ss-[(2R,3S)-3-benzyloxy-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]butyramido]-6&alpha;- aminopenclllana te The preceding 6a-methylthio derivative (0.59g, 0.83 m mole) was dissolved in dry dimethylformamide (6.4 ml) and stirred under nitrogen at-40 C.Mercuric acetate (0.26 g, I eg) and ammonia (0.016 g, 1 eq) in dimethylformamide (1 ml) were added sequentially, then the mixture was allowed to regain room temperature and stirred for 1.5 h. Workup was effected by partition of the reaction mixture between ethyl acetate and water; the organic phase was separated, washed further with water (4 x) and brine, and dried over sodium sulphate.Evaporation gave the 6a-amino penicillin as a crisp yellow foam (0.449), which was sufficiently pure to be used directly; R4O.4 in 10% methanol-chloroform; 3 (CDCI3) 1.14 (3H, t, J7 Hz, NCH2CH3), 1.26 (3H, d, J7 Hz, CH3 CHO), 1.30 and 1.42 (6H, 2s, (CH2) 2C), 2.87 (2H, brs, D2O exch, NH2), 3.30-3.75 (4H, m, 2 NCh2), 3.90-4.3Q (3H, m, NCH2 & CHNH), 4.41(1H, s, 3-H), 4.50 (1H, m, CHO), 4.59 (2H, s, PhCH2O of ether), 5.20 (2H,s, Ph CK2O of ester),5.33 (1H, s, 5-H), 7.2-7.5 (10H m, phenyls), 8.07 (1H, brs, D2O exch, 6-NH), 9.55 (1H, brd. #7 Hz, D2O exch, CHNH).
(d) Benzyl 6ss-[(2R,3S)-3-benzyloxy-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]butyramido]-6&alpha;-for- mamidopenicillanate The preceding 6&alpha;-amino derivative (0.42 g, 0.62 m mole) was dissolved in dichloromethane (5.5 ml) and stirred at 0 C while dry pyridine (0.49 ml, 10 eq) and acetic formic anhydride (0.25 ml, 5 eq) were added. The solution was allowed to regain room temperature and stirred for 1h. It was then partitioned between 0.5M, hydrochloric acid and sufficient ethyl acetate to make the organic layer the upper. The organic phase was separated and washed further with 0.5 M hydrochloric acid, water, saturated aqueous sodium hydrogen carbonate, water and brine, then dried over sodium sulphate.Evaporation gave a yellow solid which was subjected to chromatography on silica gel (40 g), eluting with 2% methanol-chloroform. The product-rich fractions, on evaporation and trituration with ether, afforded the title 6a- formamido penicillin as a white semi-solid (0.25 g, 57%); R0.35 in 10% methanol-chloro-form;; 3 [(CD2)2CO] 1.13(3H, t, J 7Hz, NCH2CH2), 1.25 (3H, d, #7 Hz, CH3CH), 1.31 and 1.46 (6H, 2s, (CH2)2C), 3.303.75 (4H, m, 2NCH2), 3.90-4.30 (3H, m, NCH, - CH NH), 4.48 (1H, s, 3-H), 4.55 (1H, m, CHO), 4.60 (2H, s, Ph CH2O of ether), 5.20 (2H, s, PhCH2O of ester), 5.60 (1H, s, 5-H), 7.20-7.50 (10H, m, phenyls), 8.12 (1H, s, NHCHO), 8.22 and 8.35 (2H, 2 brs, D2O exch, 6-NH's), 9.55 (1 H, brd, J 7 Hz, D2O exch, CHNH).
(e) 6(3-j(2h, 3S)-3-Benzyloxy-2-J(4-ethyl-2, 3-dioxopiperazin- 1-yl)carbonylamino]butyramido]-6&alpha;-formami- dopencillanic acid, sodium salt The preceding 6a-formamido benzyl ester (0.23 g; 0.32m mole) was dissolved in tetrahydrofuran: water (10 ml, 4:1). 10% Palladium on charcoal catalyst (0.15 g) was added and the mixture was hydrogenated for a total of 6h, with one filtration and change of catalyst after 3h. After this time the catalyst was filtered off and washed well with both solvents then the filtrate was concentrated to remove tetrahydrofuran and partitioned between ethyl acetate and M. sodium hydrogen carbonate solution (two portions).
The aqueous phase was acidified to pH2 with 2M hydrochloric acid and the product extracted into tetahydrofuran: ethyl acetate (3 portions, 1:1). The organic extract was dried over sodium sulphate and evaporated to dryness. The residue was dissolved in acetone and converted to its sodium salt by addition of 2M sodium 2-ethylhexanoate in methyl isobutyl ketone.This procedure afforded the title penicillin (90 mg, 55%), R, 0.45 in n-butanol:acetic acid: water, 4:1:1, p,,, (KBr) 1765,1710 cm 3 (D2O) 1.18 (3H,t,,'7 Hz, NCH2 CH3), 1,2-1.5 (9H, m, CH3CH & (CH2)2C), 3.30-3.75 (4H, m, 2 NCH2), 3.90-4.10 (2H, m, NCU2), 5.21 (2H, s, PhCH2O), 5.51(1K, s,5-H), 7.30 (5H, s, phenyls), 8.05(1K, s, NHCHO), 8.15 (1H, brd, J 7Hz, each, NH). Other signals are obscured by the water peak.
MIC ( g/ml) P. Mirabilis 889 > 100.
Example 12 Benzyl 6ss-amino-6&alpha;-formamidopenicillanate (a) Benzyl 6ss-(2,2,3-trichlorethoxycarbonylamino)-6&alpha;-(methylthio)penicillanate Benzyl 6ss-amino-6&alpha;-(methylthio)penicillanate (3.52 g, 10 mmole) was stirred at 0 C with dry pyridine (1.22 ml, 1.5 eq) in dry dichloromethane (20 ml). A solution of 2,2,2-trichloroethyl chloroformate (2.20 g, 1.5 eq) in the same solvent was added dropwise over about 0.25 h. The resulting mixture was allowed to regain room temperature, then poured into a mixture of ethyl acetate (50 ml) and 0.5 M hydrochloric acid (30 ml). The organic phase was separated and washed further with 0.5 M hydrochloric acid (2 x), water, saturated sodium hydrogen carbonate solution, water and brine, then dried over sodium sulphate.Evaporation gave the protected 6&alpha;-(methylthio) derivative as a crisp, near-colourless foam (5.17 g, 98%); R,0.80 in 10% methanol-chloroform; 3 (CDCI3) 1.45 and 1.60 (6H, 2s, C(CH3)2), 2.40 (3H, s, CH2S), 4.60(1H, s, 3-H), 4.85 (2H, narrow d, Cl3CCH2O), 5.30 (2H, s, PhCH2O), 5.60 (1H, s, 5-H), 6.40 (1H, brs, D2O exch, NH), 7.50 (5H, s, phenyls); (Found: MF, 525.9943. C19H21C13N2O5S2 requires M, 525.9940).
(b) Benzyl 6(3-(2,2,24richloroethoxycarbonylamina)-6a-aminopeniclllanate Benzyl 6ss-(2,2,2-trichloroethoxycarbonylamino)-6&alpha;-(methylthio)penicillanate (5.15 g, 9.76 mmole) was dissolved in dry dimethylformamide (20 ml) and cooled to -40 C with stirring. To this solution were added sequentially, mercuric acetate (3.15 g, 1 eq) and a solution of anhydrous ammonia (0.17 g, 1 eq) in dry dimethylformamide (6 ml). The resulting mixture was allowed to warm to room temperature over 1 h, then poured into a mixture of ethyl acetate (100 ml) and water (50 ml).The organic phase was separated and washed further with water (4 x), then with brine, and dried over sodium sulphate.Evaporation gave the 6amino penicillin as a pale yellow foam (4.44 g, 89%) which was used without further purification; R0.65 in 10% methanol-chloroform; 8 (CDCI3) 1.40 and 1.55 (6H, 2s, (CH2)2C), 2.65 (2H, brs, D2O exch, NH2), 4.55 (1H, s, 3-H), 4.80 (2H, s, Cl2CCH2O), 5.25 (2H, s, PhCH2O), 5.45 (1H, s, 5-H), 6.45 (1H, brs, D2O exch, NH), 7.45 (5H, s, phenyls).
(c) Benzyl 6ss-(2,2,2-trichlorethoxycarbonylamino0-6&alpha;-formamidopenicillanate Benzyl 6ss-(2,2,2-trichloroethoxycarbonylamino)-6a-aminopenicillanate (4.40 g, 8.86 mmole) was stirred at 0 C in dry dichloromethane (30 ml). To this solution were added dry pyridine (6.95 ml, 10 eq) and acetic formic anhydride (3.51 ml, 10 eq). The resulting pale yellow solution was allowed to warm to room temperature over 1 h, then washed sequentially with 0.5M hydrochloric acid (2 x 100 ml), water, saturated sodium hydrogen carbonate solution, water and brine and finally dried over sodium sulphate.
Evaporation gave the crude product which was subjected to chromatography on silica gel (350 g), eluting with 2% methanol-chloroform, to give the desired 6&alpha;-formamido penicillin (3.03 g 65%). This was subsequently found to crystallise on trituration with ether; m.p. 132-4"C; R,0.45 in 10% methanol-chloroform; vmax. (KBr) 3340, 3160, 1790, 1745 and 1670 cm-1; 8 (CDCI3) 1.37 and 1.53 (6H, 2s, (CH3)2C), 4.52 (1H, s, 3 H), 4.72 (2H, s, Cl2CCH2O), 5.17 (2H, s, PhCH2O), 5.66 (1H, s, 5-H), 6.75 (1H, brs, D2O exch, NH), 7.34 (5H, s, phenyls), 7.70 (1H, brs, D2O exch, NH), 8.19 (1H, s, NHCHO); Found: C, 43.6; H, 4.1; N, 7.85%.
C18K20Cl2N2O6S requires C, 43.5; H, 3.8; N, 8.0%).
(d) Benzyl 6ss-amino-6a-formamidopenicillanate Benzyl 6ss-(2,2,2-trichloroethoxycarbonylamino)-6&alpha;-formamido-penicillanate (1.44 g, 2.75 mmole) was dissolved in tetahydrofuran (45 ml) and M. potassium dihydrogen phosphate (9 ml). The resulting mixture was stirred at room temperature and powdered zinc (2 g), which had been freshly activated by washing with hydrochloric acid, then with water, was added. The pH, of the mixture was kept at 4.5 by the addition of 2M. hydrochloric acid. The progress of the reaction was monitored by t.l.c., and after about 4 h no starting material was visible. For workup the zinc was filtered off and the filtrate partitioned between ethyl acetate and water. The organic phase was separated and further washed with water and brine, then dried over sodium sulphate.Evaporation gave the crude product as a pale yellow gum (0.66 g, 84%); R0.30 in 10% methanol-chloroform. The material was not stable in this form. It could, however, be converted into its crystalline toluene-p-sulphonate salt by mixing an ethyl acetate solution with the stoichiometric quantity of toluene-p-sulphonic acid as a slurry in ethyl acetate. In this way 0.44 g (1.26 mmole) of impure free base gave 0.44 g (67%) of the 6ss-amino derivative as its toluene-p-sulphonate.
The salt had to be dissolved in d4-methanol or d6-dimethyl sulphoxide for n.m.r. purposes and rather facile decomposition ensued. However, recovery of the free base form after the salt had been standing in a desiccator for a week gave material of good purity; R, as above; 8 (CDCI2) 1.45 and 1.70 (6H, 2s, (CH2)2C); 2.60 (2H, brs, D2O exch, NHJ; 4.60 (1H, s, 3-H); 5.30 (2H, s, PhCH2O); 5.70 (1H, s, 5-H), 7.05 (1H, brs, D2O) exch, NHCHO), 7.50 (5H, s, phenyls), 8.35 (1H, s, NHCHO).
Example 13 7ss-amino-7&alpha;-formamidocephalosporanic acid, trifluoroacetic acid salt (a) t-Butyl 7a-methythio-7(3-(trichloroethoxycarbonylamino)cephalosparanate t-Butyl 7ss-amino-7&alpha;-(methylthio)cephalosporanate (6.92 g, 18.5 mmol) in dichloromethane (50 ml) with pyridine (2.2 ml, 27.2 mmol) was cooled to 0 C and treated dropwise over 0.5 h with a solution of 2,2,2trichloroethyl chloroformate (2.5 ml, 18.5 mmol) in dichloromethane (10 ml).Once addition was complete the reaction was stirred for 5 min., washed with N. hydrochloric acid, brine, dried over magnesium sulphate, and evaporated to give the almost pure product (9.86 g, 97%), 8 (CDCI3) 1.54 (9H, s, C(CH3)3), 2.06 (3H, s, OCOCH3), 2.38 (3H, s, SCH2), 3.35 & 3.49 (2H, ABq, J 18Hz, 2-H2) 4.6-5.2 (5H, m, 6-H, CH2OAc, CH2CCl2), 6.08 (1H,s, NH); vmax. (CH2C12) 3380, 2930, 1780, 1740, 1620 cm-1; (Found: M-, 548.0010.
C,6H23CI3N207S2 requires M, 548.0010).
(b) t-Butyl 7a-amino-7(3-(trichloroethaxycarbonylamino)-cephalosporanate t-Butyl 7a-methyithio-7ss-trichloroethoxycarbonylaminocephalosporanate (9.86 g, 17.9 mmol) in dimethylformamide (60 ml) at -40 C was treated with a solution of mercuric acetate (5.72 g, 17.9 mmol) in dimethylformamide (10 ml), followed by ammonia (0.30 g, 17.9 mmol) in dimethylformamide (15 ml). The reaction mixture was allowed to warm to 0 C over 1.5 h., then poured into ethyl acetate, washed well with water, brine, dried and evaporated to give the title compound (8.0 g, 86%); 8 (CDCI3) 1.56 (9H, s, C(CH3)3), 2.10 (3H, s, OCOCH2), 2.75 (2H, brs, NH2), 3.32 & 3.52 (2H, ABq, J18Hz, 2-H2), 4.4-5.2 (5H, m, 6-H, CH2OAc, CH2CCI3), 6.53 (1H, s, NH). vmax. (THF) 3200, 1790, 1740, 1730 cm--1.
(c) t-Butyl 7&alpha;-formamido-7ss-(trichloroethoxycarbonylamino)-cephalosporanate t-Butyl 7a-amino-7(3-(trichloroethoxycarbonylamino)-cephalosporanate (8.0 g, 0.015 mol) in dichloromethane (60 ml) was cooled to 0 C and treated with pyridine (13 ml, 0.16 mol) and acetic formic anhydride (6.5 ml 0.082 mol).The reaction mixture was stirred at 0 C for 1.5 h., washed with N, hydrochloric acid, saturated sodium hydrogen carbonate solution, brine, dried and evaporated to give the title compound (8.13 g, 96%); # (CDCI3) 1.53 (9H, s, C(CH3)3), 2.07 (3H, s, OCOCH2), 3.28 & 3.46 (2H, ABq, J17Hz, 2-H2), 4.7 5.3 (5H, m, 6-H, CH2OAc, CH2CCI3), 6.66 (1H, s, NH), 7.63 (1H, br s, NH), 8.22 (1H, s, CHO); v! @ (CH2C12), 3380, 1790, 1735, 1700 cm-1.
(d)-t-Butyl 7ss-amino-7&alpha;-formamidocephalosporanate t-Butyl 7&alpha;-formamdio-7ss-(trichloroethoxycarbonylamino)-cephalosporanate (8.13 g, 14.9 mmol) in tetrahydrofuran (100 ml) and M. potassium dihydrogen phosphate solution (20 ml) was stirred with zinc powder (15 g) which had been freshly activated by washing with 5N. hydrochloric acid followed by water. After 6 h. the reaction mixture was filtered and the tetrahydrofuran evaporated. The residue was diluted with ethyl acetate, washed with water, brine, dried and evaporated.The crude material was crystallized from ethyl acetate/hexane to give the product (2.5 g, 45%); m.p. 166-170 C (dec.); 8(CDCl2) 1.58 (9H, s, C(CH3)3), 2.10 (3H, s, OCOCH3), 2.43 (2H, s, NH2), 3.35 & 3.55 (2H, ABq, J18Hz, 2-Hz), 4.81 & 4.99 (2H, ABq, J13Hz, CH2OCO), 5.11 (1H, s, 6-H), 6.92 (1H, s, NH), 8.25 & 8.26 (1H, 2s, CHO); v,,,. (CH2CI2) 3410, 1790, 1740, cm-1; (Found: C, 48.5; H, 5.9; N, 11.4%. CH15H21N3O6S requires C, 48.5; H, 5.7; N, 11.3%).
(e)7ss-Amino-7&alpha;-formamidocephalosporanic acid, trifluoroacetic acid salt t-Butyl 7ss-amino-7&alpha;-amino-7&alpha;-formamidocephalosporanate (0.10 g, 0.23 mmol) in trifluoroacetic acid (5 ml) was stirred at room temperature for 0.5 h.The solution was evaporated to dryness and the residue triturated with ether and the solid product filtered and dried, (0.083 g, 72%); 8 (CF3COOH), 2.23 (3H, s, COCK2), 3.74 (2H, s, 2-H2), 5.27 and 5.41 (2H, ABq, J15Hz, CH2OCO); 5.41 (1H, s, 6-H), 8.52 (1H, s, CHO); vmax. (KBr) 3320, 2980, 2960, 1795, 1780, 1725 and 1665 cm-1.
Example 14 7ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetamido]-7&alpha;-Formamidocephalospor- anic acid, sodium salt.
(a) t-Butyl 7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7&alpha;-(methyl- thiosscephalosporanate.
D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetic acid (0.48g, 1.Smmol) in dichloromethane (30ml) was converted to its acid chloride by treatment with oxalyl chloride (0.38g, 3mmol). After stirring at room temperature for 1h, the reaction solution was evaporated to dryness.The resulting acid chloride in dichloromethane (20ml) was then added dropwise to a stirred mixture of t-butyl 7&alpha;-amino-7ss- (methylthio)cephalosporanate (0.56g, 1.Smmol) and ground 4A molecular sieves (3.0g) in dichloromethane (25ml) at 0 C. The mixture was stirred at 0-5 C for 0.5h followed by 2h at room temperature, before being filtered and evaporated to dryness.The crude product was chromatographed on silica gel 60 ( < 230 mesh ASTM) eluting with ethyl acetatelhexane 2:1 through to ethyl acetate, to afford the desired product 0.61g(60%)p,,,,,(CH2Cl2) 3390, 3280, 1790, 1740sh, 1725, 1698cm-1; 8 (CDCI2) 1.22(3H,t,J7Hz, CH2CH3), 1.53 (9H,s,C(CH3)3), 2.09(3H,s, OCOCH3), 2.30(3H,s,SCH3), 3.20 and 3.40 (2H, ABq, J 18 Hz, 2-H2), 3.54(4H,m, piperazine CH2 and CH2CH3), 3.90-4.20 (2H,m,piperazine CH2), 4.77 and 5.03 (2H,ABq, J 13Hz,CH2OCOCH3), 4.91(1H,s, 6-H) 5.67 (1H, d, J7 Hz,ot proton), 7.16 (1H,s,7ss-NHCO-), 7.30-7.55 (SH,m,aromatics)and 10.01 (1H,d,J7 Hz, a-NHCO-).
(b) t-Butyl 7&alpha;-amino-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetam- idojcephalosporanate t-Butyl 7ss-[D-2-[4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7&alpha;-(methyl- thio)cephalosporanate (0.52g, 0.77mmol) in dimethylformamide (15ml) was cooled to -50 C under nitrogen.A solution of mercuic acetate (0.25g, 0.77mmol) in dimethylformamide (0.77ml) was then added, followed immediately by ammonia (0.014g, 0.85 mmol) in dimethylformamide (1 ml). The mixture was stirred from -50 C to -20 C over a period of 1h, before being poured into ethyl acetate and washed with water and brine. The organic solution was dried over magnesium sulphate, filtered and evaporated to afford the crude product.Chromatography on silica gel 60( < 230 mesh ASTM) eluting with ethyl acetate gave the title compound (0.20g,41%); vmax. (CH2CI2) 3490, 3290, 1790, 1740sh, 1725, 1695cm-1; 8(CDCl2) 1.23(3H,t,J 6Hz, CH2 CH3), 1.54(9H,s, C(CH3)3), 2.08 (3H,s,OCOCH2), 2.95(2H, brs, NH2), 3.09 and 3.37 (2H,ABq, J 18 Hz, 2-H2), 3.54 (4H,m,piperazine CH2 and-CH2CH3), 3.88-4.20(2H,m,piperazine CH2), 4.72 and 4.97 (2H, ABq, J 13 Hz, CH20COCH3), 4.88 (1 H,s,6-H), 5.50 (1H,d#JHZ, a-proton), 7.307.50(5H,m,aromatics), 7.65 (1H, brs, 7ss-NH) and 10.00 (1H,d, J7Hz, alpha;-NHCO).
(c) t-Butyl 7ss-(D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7&alpha;-formamido- cephalosporanate.
A solution of t-Butyl 7&alpha;-amino-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetam- idol celphalosporanate (0.182g,0.29mmol) in dichloromethane (20ml) at 0 C, was treated with pyridine (0.095g, 1.2mmol) and acetic formic anhydride (0.053g, 0.6mmol).The solution was stirred at 0-5"C for 0.25h, followed by 0.75h at room temperature, It was then washed with 0.5 N. hydrochloric acid, dilute aqueous sodium hydrogen carbonate, brine, and dried over magnesium sulphate.The solution was evaporated and chromatographed on silica gel 60 ( < 230 mesh ASTM) eluting with ethyl acetate to afford the title compound (0.0599,37%);; vmax.(CHCl2) 3270, 1790, 1740sh, 1720, 1690 cm-1 8(CDCI3) 1.25(3H,t, J 7Hz, CH2CH3), 1.55(9H,s, C(CH3)3), 2.08(3H,s, OCOCH2), 2.96 and 3.28 (2H,ABq, J 18 Hz, 2-H2) 3.52(4H,m,piperazine CH2 amd-CH2CH3), 3.80-4.20 (2H, m, piperazine CH2), 4.77 and 4.99 (2H, ABq, J 13 Hz, CH2OCOH3), 5.17(1H,s,6-H), 5.53(1H,d(#7Hz, &alpha;-proton), 7.30-7.50 (5H,m,aromatics), 7.90(1H,s,7ss-NHCO), 8.16 (1H,s,7a-NHCHO), 8.19 (1H,s, NHCHO) and 10.02 (1H,d,J7Hz, a-NHCO-).
(d) 7ss-[D-2-[4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetamido]-7&alpha;-formamidocephalos- poranic acid, sodium salt t-Butyl 7ss-[D-2-[(4-ethyl-2,3-di-dioxopiperazin-1-yl) carbonylaminol-2-phenylacetamido]-7&alpha;-formamido- cephalosporanate (0.0579, 0.085 mmol) was dissolved in ice cooled 88% formic acid (5ml), and the resulting solution was stirred at room temperature for 5h. The solution was then evaporated to dryness, and toluene (5ml) was added and evaporated to ensure complete removal of the formic acid. The residue was dissolved in dilute aqueous sodium hydrogen carbonate and the resulting aqueous solution was washed with ethyl acetate.It was then saturated with sodium chloride, covered with ethyl acetateltetrahydrofuran (1:1), and acidified to pH1.5 with N. hydrochloric acid. The organic phase was separated and the aqueous phase was extracted with further ethyl acetate/tetrahydrofuran. The combined organic extracts were washed with brine, dried over magnesium sulphate, and evaporated to dryness to afford the free acid.
This was suspended in water and the Ph was adjusted to 6.5 with dilute aqueous sodium hydrogen carbonate. The resulting solution was filtered and freeze dried to afford the title compound (0.0179, 32%); vmax. (KBr0 1770, 1710, 1680, 1610, 1510cm-1; #(D2O) 1.20(3H,t,#7Hz,CH2CH3), 2.09(3H,s,OCOCO3), 3.05(1H,d,J 18 Hz, 2-H), 3.52(5H,m,piperazine CH2, CH2CH3 and 2-H), 3.80-4.10 (2H,m,piperazine CH2) 4.62 and 4.84 (2H,ABq,J13 Hz, CH20COCH3), 5.28 (lH,s,6-H), 5.52 (lH,s,a-proton),7.35-7.60 (SH,m,aromatics) and 8.15 (1H,s,NHCHO).
MIC ( g!ml) P. Mirabilis 889 0.2.
Example 15 7ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;-formamidoce- phalosporanic acid, sodium salt (a) t-Butyl 7ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino] acetamido]-7&alpha;-(methylthio)cephalosporanate D-2-[4-(Benzyloxyca rbonyloxy)phenyl]-2-[(4-ethyl-2, 3-dioxopiperazin-1 -yl)carbonylaminojacetic acid (1.88 g, 4 mmol) in tetrahydrofuran (40 ml) containing dimethylformamide (1 drop), was treated with oxalyl chloride (1.02 g, 8 mmol). After stirring at room temperature for 1.5 h the solution was evaporated to dryness, treated with toluene, and re-evaporated.The resulting acid chloride in dichloromethane (50 ml) was then added dropwise with stirring to a mixture of t-butyl 7ss-amino-7&alpha;-(methylthio)cephalosporanate (1.50 g, 4 mmol) and 4A molecular sieves (9.0 g) at 0 C. The mixture was then stirred at 0-S'C-for 0.75 h, followed by 1.75 h at room temperature, before being filtered and the filtrate evaporated to dryness.The crude product was chromatographed on silica gel 60 ( < 230 mesh ASTM) to afford an inseperable mixture of the title compound and t-butyl 7ss-[L-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxypiperazin- 1 yI)carbonylaminojacetamido]-7a-(methylthio)cephalosporante (1.07 g, 32%).
vmax (CH2C12) 3380, 3275, 1785, 1765 sh, 1740 sh, 1720, 1695, 1500, 1220 cm-1; 3 (CDCI3) 1.21 (3H, t, J 7Hz, CH2CH3), 1.54 (9H, s, C(CH3)3), 2.00 and 2.29 (3H, 2s, SCH2), 2.08 (3H, s, OCOCH,), 3.19 and 3.38, and 3.27 and 3.45 (2H, 2ABq, J 17Hz, 2-H2), 3.55 (4H, m, piperazine CH2 and -CH2CH3), 4.05 (2H, m, piperazine CH2), 4.77 and 5.05 (2H, ABq, J 13Hz, CHOCOCH3), 4.88 and 4.90 (1H, 2s, 6-H's), 5.26 (2H, s, CH2Ph), 5.64 (1H, d, J 9Hz, a-proton), 7.00-7.60 (10H, m, aromatics and 7ss-NH), and 10.02 (1H, d, #7Hz, &alpha;-NHCO).
(b) t-Butyl 7&alpha;-amino-7ss-[D-2-[4-(benzyloxycarbonyloxy)-phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]acetamido]cephalosphoranate t-Butyl 7ss-[DL-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazn-1yl)carbonylamino]acetamido-7a-(methylthio)cephalosporanate (1.07 g, 1.3 mmol) in dry dimethylformamide (25 ml) at -50 C under nitrogen, was treated with mercuric acetate (0.42 g, 1.3 mmol) in dimethylformamide (3 ml) followed immediately by ammonia (0.024 g, 1.43 mmol) in dimethylformamide (1 ml). The reaction mixture was stirred at -50 C to-20 C for 1 h, before being poured into ethyl acetate and washed well with water and brine.The organic solution was dried over magnesium sulphate, evaporated to dryness, and chromatographed on silica gel 60 ( < 230 mesh ASTM) to afford the title compound (0.29 g, 28%).
vmax (CHCl2) 3380, 3275, 1785, 1765, 1742, 1720, 1695, 1502, 1220 cm-'; 3 (CDCl2) 1.21 (3H, t, J 7Hz,-CH2CH3), 1.53 (9H, s, C(CH3)3), 2.08 (3H, s, OCOCH3), 2.60-3.20 (2H, br s, NH2), 3.04 and 3.33 (2H, ABq, J 18Hz, 2-H2), 3.65 (4H, m, piperazine CH2 and CH2CH3), 4.00 (2H, m, piperazine CH2), 4.72 and 4.98 (2H, ABq, J 13Hz, CH2OCOCH3), 4.84 (1H, s, 6-H), 5.26 (2H, s, CH2Ph), 5.57(1H, d, J 7Hz, a-proton), 7.10-7.57 (9H, m, aromatics), 7.83 (1H, s, 7ss-NHCO) and 10.05 (1H, d, J7Hz, a-NHCO-).
Also isolated from the reaction mixture was t-butyl 7a-amino-7ss-[L-2-[4-(benzyloxycarbonyloxy)phenyl]- 2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]acetamido]-cephalosporanate. (0.36 g, 35%).
vmax (CH2C12) 3380, 3275, 1785, 1765, 1740 sh, 1720, 1692,1500, 1220 cm-1; 3 (CDCl2) 1.22 (3H, t, J 7Hz, CH2CH3), 1.54 (9H, s, C(CH3)3), 2.10 (3H, s, OCOCH3), 2.50-2.80 (2H, br s, NH2), 3.19 and 3.45 (2H, ABq, J 18Hz, 2-K2), 3.55 (4H, m, piperazine CH2 and-CH2CH3), 4.02 (2H, m, piperazine CH2), 4.77 and 5.00 (2H, ABq, J 13Hz, CH2 OCOCH2), 4.88 (1H, s, 6-H), 5.27 (2H, s, CH2Ph), 5.53 (1H, d, J 7Hz, a-proton), 7.15-7.50 (10H, m, aromatics and 7ss-NHCO) and 9.94 (1H, d,J7Hz, a-NHCO).
(c) t-Butyl 7ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino] acetamido]-7&alpha;-formamidocephalosporanate A solution of t-butyl 7&alpha;-amino-7ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin 1-yl)-carbonylamino]acetamido]cephalosporanate (0.272 g, 0.343 mmol) and pyridine (0.171 g, 2.17 mmol) in dichloromethane (20 ml) was cooled in an ice bath and treated with acetic formic anhydride (0.096 g, 1.09 mmol).The reaction solution was stirred at 0-5 C for 0.5 h, and for 1 h at room temperature, before being washed with 0.5N. hydrochloric acid, dilute aqueous sodium hydrogen carbonate and brine.lt was then dried over magnesium sulphate, evaporated to dryness, and the crude product chromatographed on silica gel 60 ( < 230 mesh ASTM) to afford the title compound (0.200 g, 71%).
vmax (CH2CI2) 3375 sh, 3270, 1790, 1765, 1740 sh, 1720, 1692, 1500, 1220 cm-1; 3 (CDCl2) 1.25 (3H, t, J 7Hz, CH2 CH3), 1.54 (9H, s, C(CH3)3), 2.07 (3H, s, OCOCH3), 2.88 and 3.22 (2H, ABq, # 18Hz, 2-H2), 3.54 (3H, m,piperazine-H and-CH2CH3), 3.68-4.20 (3H, m, piperazine-H's), 4.74 and 5.00 (2H, ABq, #13Hz, CH2OCOCH3), 5.13(1H, s, 6-H), 5.26(2H, s, CH2Ph), 5.61(1H, d, # 7Hz, &alpha;-proton), 7.10-7.60 (9H, m, aromatics), 7.99 and 8.35 (2H, 2 br s, 7ss-NHCO and 7a-NHCHO), 8.16 (1H, s, NHCHO), and 10.07 (1H, d, J7Hz, &alpha;-NHCO).
(d) t-Butyl 7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)-carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;- formamidocephalosporanate A mixture of t-butyl 7P-[D-2-[4-(benzyloxyca rbonyloxy) phenyl)-2-[4-ethyl-2,3-dioxopiperazin-1- yl)carbonylamino] acetamido]-7&alpha;-fomamidocephalosporanate (0.096 g, 0.117 mmol) and 10% palladium on charcoal (0.100 9) in tetrahydrofuran (12 ml) and water (2 ml), was shaken under an atmosphere of hydrogen for 2 h. The catalyst was then filtered off, the filtrate evaporated to near dryness, and the residue dissolved in a mixture of ethyl acetateltetrahydrofuran (1:1 ).This solutidn was then washed with brine, dried over magnesium sulphate, and evaporated td dryness to leave the title compound (0.068 g, 85%).
p,,,,, (CH2C12) 3280, 1788, 1740 sh, 1718, 1690, 1515 cm-1; 3 (CDCl2) 1.20 (3H, t, J 7Hz, CH2CH3), 1.52 (9H, s, C(CH3)3), 2.08 (3H, s, OCOCH3), 3.01 and 3.30 (2H, ABq, J 18Hz, 2-H2), 3.50 (3H, m. piperazine-H and-CH2CH3), 3.60-4.20 (3H, m, piperazine-H'S), 4.74 and 4.98 (2H, ABq, J 13Hz, CH20COCH3), 5.16 (1H, s, 6-H), 5.49 (1H, d, J 13Hz, a-proton), 6.75 and 7.25 (4H, 2d, J 8Hz, aromatics), 8.10 and 8.33 (3H, 2 br s, NHCHO and 7ss-NHCO-) and 9.86 (1H, d, J 7Hz, &alpha;-NHCO).
(e) 7ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;-formam- idocephalosporanic acid, sodium salt t-Butyl 7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)-carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;-for- mamidocephalosporanate (0.047 g, 0.068 mmol) was dissolved in ice cold 98% formic acid (5 ml) and the resulting solution was stirred at room temperature for 4.5 h.The reaction solution was then evaporated to dryness, treated with toluene, and re-evaporated. The residue was dissolved in dilute aqueous sodium hydrogen carbonate and washed with ethyl acetate. The aqueous solution was saturated with sodium chloride, covered with ethyl acetateltetrahydrofurna (1:1), and acidified to pH 1.5 with N.hydrochloric acid.The organic layer was separated and the aqueous phase extracted with further ethyl acetate/tetrahydrofuran. The combined extracts were washed with brine, dried over magnesium sulphate, and evaporated to dryness to leave the free acid.This was suspended in water and the pH adjusted to 6.5 with dilute aqueous sodium hydrogen carbonate. The resulting solution was filtered and freeze dried to afford the title compound (0.018 g, 40%).
p,,,,, (KBr) 1770, 1715, 1675, 1615 cm I; # (D2O) 1.18 (3H, t, J 7Hz, CH2CH3), 2.07 (3H, s, OCOCH2), 3.08 (1H, d, J 18Hz, 2-H), 3.50 (3H, m, 2-H and CH2CH3), 3.69 (2H, m, piperazine-H2), 4.00 (2H, m, piperazine-H2), 5.27 (1H, s, 6-H), 5.40 (1H, s, a-proton), 6.90 and 7.39 (4H, 2d,J8Hz, aromatics) and 8.12 (1H, s, NHCHO).
(f) Alternative route to t-Butyl 7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxy- phenyl) acetamido]-7&alpha;-formamidocephalosporanate.
D-2-[(4-Ethyl-2,3,dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxyphenyl)acetic acid (0.335g, 1 mmol) in tetrahydrofuran (30ml), was added dropwise over 2h to a stirred solution of t-butyl 7ss-amino-7&alpha;-forman- idocephalos-poranate (0.371 g, lmmol) and N,N'-dicyclohexylcarbodiimide (0.2209, 1.1 mmol) in tetrahydrofuran (10ml). The reaction mixture was stirred for 24h at room temperature, and then evaporated to dryness.The residue was dissolved in ethyl acetate, filtered, and the filtrate evaporated to dryness. The crude product was chromatographed on silica gel 60 ( < 230 mesh ASTM) eluting with ethyl acetate through 5% ethanol/ethyl acetate, to afford the title compound (0.100g, 15%).
MIC ( g/ml) P. Mirabilis 889 0.2.
Example 16 7ss-[L-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;-formamidoce- phalosporanic acid > sodium salt.
(a) t-Butyl 7ss-[L-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1 yl)carbonylamino]acetamido]-7&alpha;-formamidocephalosporanate.
A solution of t-butyl 7&alpha;-amino-7ss-[L-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(ethyl-2,3-dioxopiperazin-1- yl)carbonylamino]acetamido]cephalosporanate (0.3509, 0.44mmol) and pyridine (0.350g, 4.4mmol) in dichloromethane (20ml) was cooled in an ice bath and treated with acetic formic anhydride (0.194g, 2.2mmol). The reaction solution was stirred at 0-5 C for 0.5h, followed by 1h at room temperature.It was then washed with 0.5N. hydrochloric acid, dilute aqueous sodium hydrogen carbonate, brine, and dried over magnesium sulphate.The solution was evaporated to dryness, and the residue chromoatographed on silica gel 60 ( < 230 mesh) to afford the title compound (0.2509, 69%); vamx(CH2Cl2), 3380, 3260, 1790, 1765, 1740sh, 1720 1695, 1500, 1220 cm-1;
; b(CDCI3) 1.24(3H,t,J7 Hz, CH2CH2), 1.53(9H,s, C(CH3)3), 2.10(3H,s, OCOCH2), 3.03(2H,s,2-H2), 3.57(3H, m, piperazine-H and-CH2CH3), 3.71, 3.89 and 4.36 (3H,3m,piperazine-H's), 4.83 and 4.16(2H, ABq J 13Hz, CH2OCOCH3), 4.98(1H,s, 6-H), 5.29(2H,s,CH2Ph), 5.91(1H,d, J 7 Hz, a-proton), 7.21 and 7.60 (4H,2d,J 8 Hz, phenyl), 7.43(5H,m,phenyl), 8.01(1H,s,NHCHO), 8.08(1H,s, NHCHO), 9.22(1H,brs, 7ss-NHCO), and 10.06(1H, d,#7 Hz, a-NHCO.
(b)-t-Butyl 7ss-[L-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamiNO]-2-(4-hydroxyphenyl)acetamido]-7&alpha;- formamidocephalosporanate.
A mixture of t-butyl 7p-[L-2-[4-(benzyloxycarbonyloxy) phenyl]-2-[(4-ethyl-2,3-dioxpiperazin-1 yl)carbonylamino] acetamido]-7&alpha;-formamidocephalosporanate(0.113g, 0.138mmol) and 10% palladium on charcoal (0.120g) in tetrahydrofuran (12ml) and water (2ml) was shaken under an atmosphere of hydro gen for 1.75h.The catalyst was then filtered off, the filtrate evaporated to near dryness, and the residue dissolved in a mixture of ethyl acetateltetrahydrofuran (1 :1).This solution was then washed with brine, dried over magnesium sulphate, and evaporated to dryness to leave the title compound (0.092g, 97%); vmax. (CH2Cl2) 3375, 3290, 1790, 1740sh, 1715, 1695, 1515cm-1; 8[(CD3)2CO] 1.18(3H, t,J7 Hz, CH2CH3), 1.52(9H,s, C(CH3)3), 2.03 (3H,s, OCOCH3), 3.20-3.87(6H,m, piperazine-H2, 2-H2, and-CH2CH3), 3.904.30(2H,m,piperazine-H2), 4.75 and 5.05 (2H,ABq, J 13 Hz, CH21OCOCH2), 5.26(1H,s,6-H), 5.75 (1H,d,J7 Hz, a-proton), 6.89 and 7.40 (4H, 2d, J 8 Hz, aromatics). 8.20 (1H,s, NHCHO) 8.38 (1H,s, NHCHO), 8.95(1H,brs, 7ss-NHCO) and 10.00 (1H,d,#7 Hz, &alpha;-NHCO).
(c) 7ss-[L-2-[(4-Ethyl-2,3-dioxopiperazin- l-yl) carbon ylamino]-2-/4-hydroxyphen Vl)acetamido]- 7a-for- mamidocephalosporanic acid, sodium salt t-Butyl, 7ss-[L-2-[(4-ethyl-2,3,dioxopiperazin-1-yl) carbonylamino]-2-(4-hydroxyphenyl)acetamido]-7&alpha;-for- mamidocephalos-poranate (0.0899, 0.129 mmol) was dissolved in ice cooled 98% formic acid (10ml), and the resulting solution was stirred at room temperature for 5h. The reaction solution was then evaporated to dryness, treated with toluene, and re-evaporated.The residue was dissolved in dilute aqueous sodium hydrogen carbonate and washed with ethyl acetate.The aqueous solution was saturated with sodium chloride, covered with ethyl acetate/tetrahydrofuran (1:1), and acidified to pH 1.5 with N. hydrochloric acid. The organic layer was separated and the aqueous phase extracted with further ethyl acetate/tetrahydrofuran. The combined extracts were washed with brine, dried over magnesium sulphate, and evaporated to dryness to leave the free acid. This was suspended in water and pH adjusted to 6.5 with dilute aqueous sodium hydrogen carbonate.The resulting solution was filtered and freeze dried to afford the title compound (0.020g, 24%); v,,,. (KBr) 1770, 1710, 1675, 1615, 1515 cm (D2O) 1.16(3H,t, J7 Hz, CH2CH3), 2.07 (3H,s, OCOCH2), 3.04(1H,d, J 18 Hz, 2-H), 3.47 (3H,m, 2-H and CH2CH3), 3.66(2H,m, piperazine-H2), 3.98(2K, m, piperazine-H2), 4.64(1H,d,J 13 Hz, CHOCOCH3), 5.19(1H,s, 6-H, 5.42(1H,s, a-proton), 6.89 and 7.33(4H,2d,J 8 Hz, aromatics) and 8.06 (1 K,s, NHCHO).
MIC (pg/ml) P. Mirabilis 889 100.
Example 17 7&alpha;-Formamido-7ss-(thien-2-ylacetamido)cephalosporanic acid, sodium salt.
(a) t-Butyl 7ss-(thien-2-ylacetamido)-7&alpha;-(methylthio)cephalosporanate Thiophene-2-acetic acid (0.85g , 6.0mmol) was refluxed in thionyl chloride (5ml) for lh.Excess thionyl chloride was evaporated, and the residual acid chloride in dichloromethane (10ml) added to an ice cooled solution of t-butyl 7ss-amino-7&alpha;-(methylthio)cephalosporanate (1.809, 4.8 mmol) in dichloromethane (25ml) with pyridine (0.57ml), 7.0mmol). The reaction mixture was stirred at room temperature for 2h.
The solvent was evaporated and the residue taken up in ethylacetate, washed with N. hydro chloricacid,saturated sodium hydrogen carbonate solution, brine, dried over magnesium sulphate, and evaporated. Chromatography (silica gel, 3:1 hexane/ethyl acetate) afforded the title compound (1.759, 73%); #(CDCl3) 1.51(9H,s, C(CH3)3, 2.04(3H,s, OCOCH3), 2.24(3H,s,SCH3), 3.30, 3.42 (2H, ABq, # 18Hz, 2-H2), 3.84 (2H, s, ArCH2), 4.84, 5.05 (2H, ABq, J 13.5 Hz, CH2O), 4.89(1H,s, 6-H), 6.46 (1H,s, NH), 6.92-7.04 (2H, m, thiophene-H's), 7.16-7.30 (1H,m, thiophene-H); vmax (CK2CI2), 3280, 2970, 2920, 1775, 1735 sh, 1720, 1670, 1510 cm-1; (Found: M+, 498.0930. C21H25N2O6S3 requires M, 498.0952).
(b) t-Butyl 701-amino-7P-lthien-2-ylacetamidoj cephalosporanate.
t-Butyl 7(3-(thien-2-ylacetamido)-7a-(methylthio)cephalosporanate (1.00g, 2.0mmol) in dimethylformamide (20ml) at -40 C,was treated with a solution of mercuric acetate (0.649, 2.0mmol) in dimethylformamide (5ml) followed by a solution of ammonia (0.0349, 2.0mmol) in dimethylformamide (1.5ml). The reaction mixture was allowed to warm over 1.5h, poured into ethyl acetate, washed well with water, brine,dried over magnesium sulphate and evaporated to give the desired product (0.90g, 96%);; b(CDCI3) 1.54(9H, s, C(CH3)3), 2.09(3H,s, OCOCH3) 2.81(2H,brs, NH2) 3.25, 3.55 (2H,ABq, J 18 Hz, 2-H2), 3.90(2H,s, ArCH2), 4.5-5.3 (3H,m, 6-H and CH2O), 6.87-7.4(3H,m, aromatic); vmax. (tetrahydrofuran) 1790, 1745, 1725 & BR< 1680cm-1.
(c) t-Butyl 7ct-formamido-7ss-{thien-2-ylacetamido)cephalosporanate.
t-Butyl 7&alpha;-amino-7ss-(thien-2-ylacetamido)cephalosporanate (0.90g, 1 .9mmol) in dichloromethane(20ml) at 0 C was treated with pyridine (1.51 ml, 19mmol) and acetic formic anhydride (0.76, 9.5mmol). The reaction mixture was stirred at 0 C for 1h, washed with N. hydrochloric acid, saturated sodium hydrogen carbonate solution, brine, dried over magnesium sulphate and evaporated.The product was recrystallised from dichloromethane/hexane m.p. 160-4 C (0.629,66%); b((CD3)2CO) 1.54(9H,s,C(CH3) 2.00 (3H,s, OCOCH2), 3.33, 3.61(2H,ABq,J 18 Hz, 2-H2), 3.92(2H,s,ArCH2), 4.73, 4.99(2H,ABq,J 13 Hz, CH2O), 5.21 (1H,6, 6-H) 6.8-7.1 (2H,m,thienyl-H'S)7.2-7.4 (1H,m, thienyl-H), 8.19(1H,s,CHO); vmax (KBr) 3330,2980, 1770, 1740, 1720, 1695, 1660 & 1530 cm-1, (Found: C,50.7; H,5.1; N,8.4%. C21H25N2O7S2 requres C, 50.9; H,5.1; N,8.5%) (d) 7et-Formamido-7ss-{thien-2-ylacetamido)cephalosporanic acid, sodium salt.
t-Butyl 7a-formamido-7(3-(thien-2-ylacetamido)cephalosporanate (0.1009, 0.2mmol) was added to ice cold 98% formac acid (8ml) and water (5 drops). The reaction mixture was stirred at room temperature for 4h. Formic acid was evaporated and the residue taken up in dilute sodium hydrogen carbonate solution and washed with ethyl acetate. The aqueous phase was acidified to pH 1.5 and extracted with ethylacetate. The extracts were washed with brine, dried and evaporated.The residue was taken up in water, which was adjusted to pH 6.5 with sodium hydrogen carbonate solution. The solution was filtered and freeze dried to give the title compound (0.054g 58%); 3(D2O) 2.09(3H,s,OCH3), 3.23,3.57 (2H,ABq, J 17 Hz, 2-H2), 3.92(2H,s, ArCH2), 5.28 (2H,s, CH2O), 5.60(1H,s,6-H), 6.9-7.2(2H,m,thiophene-H's), 7.37.5(1H,m,thiophene-H), 8.14(1H,s,CHO); vmax (KBr) 1770, i740, 1720, 1695, & 1660 cm--1.
MIC ( g/ml) P. Mirabilis 889 10.
Example 18 7ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenyl-acetamido]-7&alpha;-formamido-3-methyl-1- oxadethia-ceph-3-em-4-carboxylic acid.
(a) t-Butyl 7ss-(4-nitrobenzylideneamino)-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate A solution of t-butyl 7p-amino-3-methyl-l -oxa-dethia-ceph-3-em-4-ca rboxylate (C.L. Branch and M.J.
Pearson; J. Chem. Soc. Perkin Trans 1. 1979; 2268) (186mg; 0.73 m mol) and 4-nitrobenzaldehyde (110 mg; 0.73 m mol) in an anhydrous mixture of toluene (25 ml) and ethyl acetate (5 ml) was stirred for 6 h at room temperature over 4A molecular sieves. The reaction mixture was then filtered and the filtrate evaporated to dryness.The residue was redissolved in dry toluene and the solution evaporated; this was repeated. Trituration of the residue with ether gave the title compound as a white solid (227 mg; 80%); vmax. (CHCl3) 1780, 1715, 1640, 1525, 1350 cm-1; b(CDCI3) 1.56 (9H, s), 2.01 (3H, s), 4.36 (1H, s), 5.31 (2H, s), 7.98 (2H, d, J 8Hz), 8.29 (2H, d, J 8Hz), 8.78 (1H, s).
(b) t-Butyl 7ss-(4-nitrobenzylideneamino)-7&alpha;-methylthio 3-methyl-1-oxadethia-ceph-3-em-4-carboxylate t-Butyl 7p-(4-nitrobenzylideneamino)-3-methyl-l -oxadethia-ceph-3-em-4-carboxylate (257 mg; 0.66 m mol) was dissolved in anhydreus methylene chloride (5 ml) containing methyl methanethiol sulphonate (92 mg; 0.73 m mol) and cooled to 0 C under argon. To the vigorously stirred solution was added dropwise 1,8-diazabicycle [5.4.0] undec-7-ene (100 mg; 0.66 m mol) in methylene dichloride (2 ml).After 1 h at 0-5 c, the solution was diluted with methylene chloride and washed successively with saturated aqueous ammonia chloride (x2), brine, dried (MgSO4), and evaporated. Chromatography of the residue on silica gel afforded the title compound as a pale yellow solid (242 mg; 84%); vmax. (Nujol) 1740, 1620 cm-1; 8(CDCl2) 1.57 (9H, s), 1.99 (3H, s), 2.30 (3H, s), 4.33 (2H, s), 5.14 (1H, s), 8.02 (2H, d, J 8Hz), 8.29 (2H, d, J 8Hz), 8.87 (1 H, s).
(c) t-Butyl 7ss-amino-7&alpha;-methylthio-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate, toluene-ss-sulphonic acid salt t-Butyl 7(3-(4-nitrebenzylideneamino)-7a-methylthie-3-methyl-1 -oxadeth ia-ceph-3-em-4-ca rboxylate (238 mg; 0.54 m mol) was dissolved in a mixture of ethyl acetate (8 ml) and methylene dichloride (4 ml) and a solution of toluene-p-sulphonic acid monohydrate (105 mg; 0.55 m mol) in a little ethyl acetate added.
Precipitation occurred almost immediately and after stirring at room temperature for 2h, the product was filtered off, washed well with ethyl acetate, ether, and dried in vacuo to afford the title product as a white solid (193 mg; 75%); uma. (Nujol) 3150, 1780, 1715, 1705, 1640 cm-1; #[(CD3)2SO] 1.47 (9H, s), 1.94 (3H, s), 2.30 (3H, s), 2.39 (3H, s), 4.49 (2H, AA'), 5.35 (1H, s), 7.10 (2H, d, J 9Hz), 7.47 (2H, d, J 9Hz), 8.0-9.7 (3H, br, exch.).
(d) t-Butyl 7(3-fD-2-(4-ethVl-2, 3-dioxopiperazin- 1-yl) carbonylamino]-2-phenylacetamido-7&alpha;-methylthio- 3-methyl-1-oxadethia-ceph-3-em-4-carboxylate D-2-[(4-Ethyl-2-, 3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetic acid (200 mg; 0.63 m mol) in anhydrous methylene chloride (5 ml) containing a catalytic amount of dimethylformamide, was treated with oxalyl chloride (174mg; 1.34 m mol). After stirring at room temperature for 1.5h the solution was evaporated to dryness, treated with toluene and re-evaporated.The resulting acid chloride was taken up in dry methylene chloride (5 ml) and added dropwise with stirring to a mixture of t-butyl 7ss-amino-7&alpha;-methyl- thio-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate, toluene-p-sulphonic acid salt (193 mg; 0.41 m mol), and pyridine (84 mg; 0.94 m mol) in methylene chloride (5 ml) at 0 C. After stirring at 0-5 C for 0.5h and 1h at 5'C-rnom4emperature, the reaction mixture was washed with dilute hydrochloric acid, saturated aqueous sodium hydrogencarbonate, brine, dried (MgSO4), and evaporated.Chromatography of the residue on silica gel gave the title compound as an amorphous solid (169 mg; 67%); v,,,. (CHCI2) 3400, 3275, 1785, 1720 sh, 1710, 1690, 1640 sh cm-1; ô(CDCI3) 1.20 (3H, t, J 8Hz), 1.51 (9H, s), 1.93 (3H, s), 2.28 (3H,s), 3.4-3.7 (4H, m), 3.9-4.3 (2H, m) overlapping 4.03 and 4.19 (2H, ABq, J 17Hz), 4.93 (1H, s), 5.55 (1H, d, J 6Hz), 6.52 (1H, s), 7.3-7.5 (5H, m), 9.89 (1H, d, J 6Hz).
(e) t-Butyl 7c-Amino-7ss-[D-2-[{4ethyl-2-, 3-dioxopiperazin-1-yl carbonylamino]-2-phenylacetamido] 3methyl-1-oxadethia-ceph-3-em-4-carboxylate To a vigorously stirred solution of t-Butyl 7ss-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1-yl) carbonylamino]-2 phenylacetamido]-7&alpha;-methylthio-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate (30 mg; 0.05 m mol) in dioxan (2 ml)at room temperature was added peracetic acid (75 ml of 5.07% solution in acetic acid; 0.05 m mol). After 15 min. the solution was evaporated to dryness. The residue was taken up in dry toluene and the solution evaporated; this was repeated twice.Chromatography of the residue on silica gel gave t-Butyl 7(3-{D-2-[(4-ethyl-2, 3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7es-methylsulphinyl- 3-methyl-1-oxadethia-ceph-3-em-4-carboxylate as a white solid (29 mg; 94%), which was dissolved in anhydrous tetrahydrofuran (2 ml) and ammonia (2.3 ml; 0.10 m mol) added.The reaction mixture was left overnight at room temperature, evaporated and the residue chromatographed on silica gel to afford an inseparable mixture of the title compound; vmax (CHCI3) 3400, 3275, 1780, 1715, 1690, 1640 sh cm-1; 5 (CDCI3) inter alia 1.22 (3H, t, J 6Hz), 1.52 (9H, s), 1.94 (3H, s), 2.5-2.9 (2H, br s, exch.) 3.4-3.7 (4H, m), 3.9-4.2 (2H, m), 4.02 and 4.22 (2H, ABq, J 18Hz), 4.87 (1H, s), 5.42 (1H, d, J 6.5 Hz), 6.57 (1H, s exch.), 7.37.5 (5H, s), 9.92 (1H, d, J 6.5Hz); and t-butyl 7ss-amino-7&alpha;-[D-2-[4-ethyl-2, 3-dioxopiperazin-1-yi) carbony lamino]-2-phenylacetamido] 3-methyl-l -oxadethia-ceph-3-em-4-ca rboxylate (20 mg; 75%); 5 (CDCI3) inter alia 1.22 (3H, t, J 6Hz), 1.52 (9H, s), 1.95 (3H, s), 2.5-2.9 (2H, br s, exch.); 3.4-3.7 (4H, m), 3.9-4.2 (2H, m), 4.40 (2H, s), 5.03 (1H, s), 5.42 (1H, d, J 6.5 Hz), 6.59 (1H, s, exch.), 7.3-7.5 (5K, s), 9.92 (1H, d, 6.5 Hz).
(f) Alternative route to t-Butyl 7&alpha;-amino-7ss-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1-yl) carbonylamino]-2phenylacetamido]-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate t-Butyl 7ss-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1 -yl) carbo nylamino]-2-phenylacetamido]-7ssmethylthio-3- methyl-1-oxadethia-ceph-3-em-4-carboxylate (30 mg; 0.05 m mol) in dry dimethylformamide (0.5 ml) at -50 C under argon, was treated with mercuric acetate (16 mg; 0.05 m mol) in dimethylformamide (0.1 ml) followed immediately by ammonia (17 mg; 0.08 m mol) in dimethylformamide (0.1 ml).The reaction mixture was stirred at -50dg to -20dgC for 1h, before being poured into ethyl acetate and washed well with water and brine.The organic solution was dried over magnesium sulphate, evaporated, and the residue chromatographed on silica gel to afford the title product and t-butyl 7ss-amino-7&alpha;-[D-2-[(4-ethyl-2, 3dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-3-methyl-1-oxadithiaceph-3-em-4-carboxylate as an inseparable mixture (18 mg; 63%).
(g) t-Butyl 7ss[D-2-[(4-ethyl-2-, 3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7&alpha;-formamido 3-methyl- 1-oxadethia-ceph-3-em-4-carboxylate A solution of the mixed C-7 isomers of t-Butyl 7-amino-7-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-3-methyl-1-oxadethia-ceph-3-em-3-carboxylate (80 mg; 0.14 m mol) and pyridine (109 mg; 1.4 m mol) in methylene dichlpride (3 ml) was cooled to 0 C and treated with acetic formic anhydride (62 mg; 0.7 m mol). The reaction was stirred at 0-5 C for 0.5h and for 1h at room temperature before being diluted with dry toluene and evaporated. The residue was redissolved in dry toluene and the solution evaporated; this was repeated twice.Chromatography of the residue on silica gel gave the title product as an amorphous solid (45 mg; 54%); Amax.(EtOH) 257 n m (@12211), vmax. (CHCI3) 3400, 3275, 1790, 1715, 1690, 1640 sh cm-1; 5 (CDCI3) 1.20 (3H, t, J 7Hz), 1.50 (9H, s), 1.92 (3H, s), 3.4-3.7 (4H, m), 3.9-4.3 (2H, m) overlaps 4.08 and 4.23 (2H, ABq, J 17Hz), 5.14 (1H, s), 5.49 (1H, d, J 6Hz), 7.2-7.5 (6H, m), 7.57 (1H, s, exch.), 8.15 and 8.49 (s and d, J 13 Hz, together 1H, collapses to 2 s at 8.15 and 8.49 on exch.), 9.90 (1 H, d, J 6Hz).Also isolated from the reaction mixture was t-butyl 7&alpha;-[D-2-[(4-ethyl-2, 3dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7ss-formamido-3-methyl-1-oxadethia-ceph-3-em4-carboxylate (6 mg; 7%); vmax, (CHCI2) 3400, 3275, 1790, 1715, 1690, 1640 sh cm-'; b(CDCI3) inter alia 1.20 (3H, t, J 7Hz), 1.50 (9H, s), 1.94 (3H, s), 3.4-3.8 (4H, m), 3.9-4.2 (2H, m), 4.31 (2H, s), 5.08(1H, s), 5.5(1H, d, J 6Hz), 7.2-7.5 (6H, m), 7.88 (1H, exch.), 8.05 and 8.27 (s and d, J 13Hz, together 1H; collapses to 2s, at 8.05 and 8.27 on exch.), 9.92 (1H, d, J 6Hz).
(h) 7ss-[D-2-[(4-Ethyl-2, 3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]-7&alpha;-formamido-3- methyl-1-oxadethia-ceph-3-em-4-carbonylic acid t-Butyl 7ss-[D-2-[(4-Ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylactamido]-7&alpha;-formamido-3- methyl-1-oxadethia-ceph-3-em-4-carboxylate was briefly treated with trifluoroacetic acid to afford the title compound as an off-white solid; vmax. (KBr) 3400-3300 br, 1785, 1710, 1680 cm-1.
Example 19 6&alpha;-Formamido-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate, disodium salt (i) Benzyl 6&alpha;-amino-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate, triethylammonium salt Benzyl 6&alpha;-methylthio-6ss-(R-2-phenyl-2-sulphoacetamdio) penicillanate triethylammonium salt (3.259) in dimethylformamide (15ml) at room temperature was treated with mercuric acetate (1.60g) followed after 1 minute by a solution of ammonium acetate (0.779) and triethylamine (1.05ml) in DMF (10ml). After 10 minutes chloroform (250ml) and 0.5M aqueous triethylammonium sulphide (15ml, pH7.3) were added, the mixture filtered, the aqueous layer discarded and the chloroform solution dried over anhydreus magnesium sulphate then evaporated to an oil in vacuo. The oil was triturated with ether (2 x 100ml) then acetone (20ml) added and set aside to crystallise. The crystals (1.679) were collected, washed with acetone, then ether and dried in vacuo.; vmax (KBr) 1780, 1740, 1670, 1535, 1325, 1245, 1205, 1175 and 1040cm-1; #[(CD3)2SO] 1.15 (9H, t, J7Hz, HN(CH2CH3)3) 1.27, 1.40(6H, 2s, 2-(CH3)2), 3.06(6H,q,#7Hz, HN(CH2CH3)3,), 3.23 (2H, brs, NH2), 4.45 (1H,s3-H), 4.51(1H,s,CHCONH), 5.18 (2H,s,OCH2Ph0, 5.21(1H,s,S-K), 7.1-7.5 (10H,m,phenyls), 9.27 (1H,s,CONH).
(ii) Benzyl 6&alpha;-formamido-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate, triethylammonium salt Benzyl 6&alpha;-amino-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate, triethylammonium salt (0.62g) in dichloromethane (5ml) was treated with acetic formic anhydride (0.2ml) followed by pyridine (1 ml).After 5 minutes at room temperature the solution was poured into ether (100ml) and the precipitated solid collected, washed with ether and dried in vacuo to give the title compound (0.49g, 75.6%); vmax (KBr) 1785, 1742, 1675, 1320, 1245, 1205, 1180, and 1040cm-1; #[(CD3)2CD] 1.20 (9H,t,J7Hz, N+H(CH2CH3)3), 1.30, 1.48 (6H, 2s, 2-(CH2)2), 3.08 (6H, q, J 7Hz, N+H(CH2CH3)3), 4.49 (1H,s,3-H), 4.98 (1H,s,CHCONH), 5.21 (2H,s,OCH2Ph), 5.68 (1H,s,5-H), 7.27-7.8(10H,m,phenyls), 8.04 (1H,s,CONH), 8.17(1H,s,NHCHO), 8.4-9.2 (1H,brs,N+H), 9.38 (1H,s,CONH).
(iii) 6&alpha;-Formamido-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate, disadium salt Benzyl 6&alpha;-formamido-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanate triethylammonium salt (0.309) in water (10ml) containing N sodium hydrogen carbonate (0.93ml) was hydrogenated in the presence of 10% palladium on carbon catalyst (0.39) for 0.5h. The catalyst was filtered off and the filtrate passed through an Amberlite IR120 (Na) column, then freeze dried (0.22g, 94%); vmax (KBr) 1772, 1675, 1610, 1210 and 1042cm-1; 8 (D2O) 1.09, 1.28 (6H,2s,2-(CH2)2), 4.15 (1H,s,3-H), 5.05 (1H,s,CHCONH), 5.56 (1H,s,S-H), 7.3-7.7 (5H,m,Ph), 8.10(1 K,s,NKCHO).
Example 20 Benzyl 6ss-[D-2-[4-benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1 yl)carbonylamino]acetamidol-6a-formamidopenicillanate (a) Benzyl 6&alpha;-amino-6ss-[D-2-[4-(benzyloxycarbonyloxy) phenyl]-2-(2,2,2-trichloroethoxycarbonylam ino)acetamido] penicillanate A solution of benzyl 6ss-[D-2-[4-(benzyloxycarbonyloxy) phenyl]-2-(2,2,2-trichloroethoxycarbonylam ino)acetamido]-6a-(methylthio)pencillan (0.460 g) in dry N, N-dimethyl-formamide (7 ml) was stirred under nitrogen and cooled to -40 C. A solution of mercuric acetate (0.81 g) in N, N-dimethylformamide (1 ml) was added, followed by ammonia (0.011 g) in N, N-dimethylformamide (0.58 ml).The reaction mixture was stirred at -50' to -10 C for 1h and then partitioned between ethyl acetate and water. The organic phase was washed four times with water, once with brine, dried dver sodium sulphate and concentrated.Chromatography of the residue on silica gel 60 (230 mesh ASTM), eluting with ethyl acetate/hexane 1:1 gave benzyl 6&alpha;-amino-6ss-D-2-[4-benzyloxycarbonyloxy)phenyl]-2-(2,2,2-trichloro-ethoxy- carbonylamino)acetamido penicillanate (0.285 g); v max. (CHCl3) 3400, 3310, 3020, 2950, 1765, 1745, 1685, 1605, and 1500 cm-l; b(CDCI3) 1.00 and 1.20 (6H, 2s, 2-CH3's), 2.60(2H, s, NH2): 4.30 (1H, s, 3-H), 4.64 (2H, s,CH2CCI3), 5.11 and 5.20 (4H, 2s, PhCK2,s), 5.24(1K, d, J 7Hz, NCHCO), 5.36 (1H, s,5-H), 6.41(1H, d, J 7 Hz, NH), 7.10 and 7.44 (4H, 2d,J8 Hz, phenyl), 7.31 and 7.36 (11H, 2s, phenyl and NH).
(b) Benzyl 6ss-fD-2-[4-(benzyloxycarbon yloxy)phenyl]-2-(2,2,2-trichloroethoxycarbonylamino)acetamido]- 6a-formamidopencillanate A solution of benzyl 6&alpha;-amino-6ss-[D-2-4-benzyloxycarbonyloxy0phenyl]-2-(2,2,2-trichloroethoxycarbon- ylamino) acetamido]pencillanate (0.256 g) in dichloromethane (10 ml) was stirred under argon and cooled to 0 C.
It was treated with pyridine (0.26 ml) and acetic formic anhydride (0.13 ml), and then allowed to warm to room temperature over a period of 3h. The resulting solution was washed with 0.5N hydrochloric acid, dilute aqueous sodium < hydrogen carbonate, brine and dried over sodium sulphate. The solution was concentrated and chromatographed on silica gel 60 ( < 230 mesh ASTM) eluting with ethyl acetate/hexane 1:1 to give benzyl 6ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-(2,2,2-trichloroethoxycabonylam.
ino)acetamido]-6a-form- amidopencillanate (0.230 g) as a colourless foam; v max (CHCI3), 34200, 3300, 3000 br, 1780 sh, 1765,1745, 1695, 1605 and 1500 cm-1; 8 (CDCI3) 1.00 and 1.20 (6H, 2s, C(CH3)2), 4.36 (1H, s, 3-H), 4.64 (2H, s, CH2 CCI2), 5.12 and 5.20 (4H, 2s, PhCH2,s), 5.48 (1H, d,J7 Hz, NCHCO), 5.56 (1H, s, 5-H), 6.56 (1H,brd, J 7 Hz, NH), 7.09 and 7.47 (4H, 2d, J 9 Hz, phenyl), 7.30 and 7.37 (11H, 2s, phenyls and NH), 8.09 (1H, s, NCHO), and 8.50 (1H, b, NH).
(c) Benzyl 6ss-[D-2-amino-2-[4-(benzyloxycarbonyloxy)phenyl]-acetamido]-6&alpha;-formidopencillanate A solution of benzyl 6ss-[D-2-[4-(benzyloxycarbonyloxy)-phenyl]-2-(2,2,2-trichloroethoxycarbonylam- ino)acetamido]-6&alpha;-formamidopencillanate (0.200 g) in tetrahydrofuran (10 ml) was stirred at room temperature and treated with M. aqueous potassium hydrogen phosphate (2 ml) followed by freshly acid washed zinc powder (0.400 g). Stirring was continued until deprotection was complete (ca 4 h) and the mixture was then filtered through celite. The filtrate was concentrated and partitioned between ethyl acetate and brine.The ethyl acetate solution was dried over sodium sulphate and concentrated to give benzyl 6ss-[D-2-amino-2-[4-(benzyloxycarbonyloxy)-phenyl]acetamido]-6&alpha;-formamidopencillanate.
(d) Benzyl 6ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopiperazin-1 yl)carbonylamino]acetamido]-6&alpha;-formamidopencillanate.
Benzyl 6ss-[D-2-amino-2-[4-(benzyloxycarbonyloxy)phenyl] acetamido]-6a-formamidopencillanate was prepared from the corresponding N-trichloroethoxycarbonyl derivative (0.200 g) and then dissolved in dry tetrahydrofuran (5 ml). It was stirred at 0 C under nitrogen and treated with 4-ethyl-2,3-dioxopiperazine-1- carbonyl chloride (0.050 g).The reaction mixture was allowed to warm to room temperature over a period of 1h and then concentrated. The residue was taken up in ethyl acetate, washed with brine, dried over sodium sulphate and concentrated.The residue was chromatographed on silica gel 60 ( < 230 mesh ASTM) to give benzyl 6ss-[D-2-[4-(benzyloxycarbonyloxy)phenyl]-2-[(4-ethyl-2,3-dioxopipefrazin-1- yl)carbonylamino] acetamido]-6ol-formamidopenicillanate (0.055 g); p,,,, (CHCI3) 3280, 2980, 2875, 1780, 1765, 1745, 1710, 1685, 1610 and 1505cm-1; 8 (CDCI3) 0.85 and 1.12 (6H, 2s, 2-CK2,s), 1.26 (3H, t,#7Hz, CH3 of NEt), 3.4-3.8 (6H, m, NCH2CH2NCH2), 4.36 (1H, s, 3-H), 5.07 and 5.16 (2H, ABq, J 12 Hz, benzylic CH2), 5.21 (2H, s, benzylic CH2), 5.50 (1H, s, 5-H), 5.65 (1H, d, J 7Hz, NCHCO), 7.11 and 7.53 (4H, 2d, J 9 Hz, phenyl), 7.2-7.5(10H, m, phenyls), 8.05 and 8.69 (2H, 2 br s, NH's), 8.14 (1H, d,,l7 Hz, NH).
Example 21 GP-ph en ox yaceta mido-6oc-fo rmamidop enam3-carboxylic acid (a) Benzyl 6ss-(4-nitrobenzylideneamino)penam-3-carboxyate Benzyl 6ss-aminopenam-3-carboxylate (1.11g, 4 m mol) in dry dichloromethane (4ml) with trimethylorthoformate (0.42g, 0.44 ml, 4 m mol) was treated with 4-nitrobenzaldehyde (0.61g, 0.4 m mol) in dry methanol (10 ml) at room temperature for 1.5h. The mixture was then evaporated to dryness, triturated with ethanol and the solid material collected by filtration.Recystallisation from ethanol/ethylacetate gave almost colourless needles (1.2g 74%); m.p. 95-96C; vmax (CH2CI2) 1795, 1750, 1530, 1360 cm-1; 8(CDCI3) 8.68 (1H, d, J 2Hz, CH=N), 8.27 and 7.94 (4H, 2d, J 9Hz, nitrephenyl), 7.38 (5H, s, phenyl), 5,51(1K, d, J 4Hz, 5-H), 5.45 (1H, dd, J 2Hz and 4 Hz, 6-H), 5.23 (2H, s, CHPh), 4.93 (1H, t, J 5.5Hz, 3-H) and 3.45 (2H, d, J 5.5Hz, 2-H).
(b) Benzyl 6ss-amino-6&alpha;-(methylthio)penam-3-carboxylate Benzyl 6ss-(4-nitrobenzylidenamino) penam-3-carbexylate (1.05 g, 2.55 m mol) in dry ethylacetate (30 ml) was cooled to -10 C and methyl methanethiolsulphonate (0.32 g, 0.26 ml, 2.55 m mol) followed by potassium hydroxide (0.16g, 2.55 m mol) in ethanol (5 ml) were added. After the addition the mixture was stirred for 0.5h, poured into water (25 ml) and the organic phase separated, washed with water (2 x 25 ml), brine (2 x 25 ml) then dried (Mg SO4) and finally concentrated to ca 10 ml. This solution was treated with toluene-p-sulphonic acid monohydrate (0.48 g, 2.55 m mol) and stirred at room temperature for 3 h. The solution was then washed with dilute sodium hydrogen carbonate solution, brine, dried and evaporated to give an orange gum which was chromatographed on silica.gel 60 (230 mesh ASTM) to give the title compound as a yellow gum (0.41g, 49%); vma. (CH2Cl2) 1795, 1750 cm-1; (CDCI3) 7.45 (5H, s, phenyl), 5.27 (3H, s, 5-H and CHPh), 5.15 (1H, m, 3-H), 3.45-3.25 (2H, m, 2-H2), 2.31 (3H, s, SCH30 and 2.18 (2H, rs, NH2).
(c) Benzyl 6(3-phenoxyacetamido-6a-(i eth ylthio)penam-3-carboxyla te Benzyl 6ss-amino-6&alpha;-(methylthio) penam-3-carboxylate (0.11g, 0.34 m mol) in dry dichloromethane (5 ml) at 0 C was treated with pyridine (0.035g, 0.036 ml, 1.3 eq.) and then with phenoxyacetyl chloride (0.058g, 0.038 ml, 0.34 m mol).The mixture was allowed to warm to room temperature and stirred for 2h, when it was washed with dilute hydrochloric acid (10 ml), dilute sodium hydrogen carbonate solution (10 ml) water (10 ml), brine (10 ml), dried (MgSO4), and evaporated to give an orange gum.This crude material was chromatographed on silica gel 60 ( < 230 mesh ASTM) to give the title compound as a pale yellow foarn (0.026g, 54%); vmax. (CH2Cl2) 1800, 1755, 1700, 1500 cm-1; #(CDCl3) 7.41-6.92 (1H, m, aryl-H's and NH), 5.32 (1H, s, 5-H), 5.20 (2H, ABq, CH2Ph), 5.08 (1H, dd, J 6 and 2 Hz, 3-H), 4.57 (2H, s, PhOCH2), 3.42 (2H, m, 2-H2) and 2.32 (3H, s, SCK2).
(d) Benzyl 6ss-phenoxyacetamido-6&alpha;-aminopenam-3-carboxylate Benzyl 6ss-phenoxyacetamido-6&alpha;(methylthio) penam-3-carbexylate (0.048g, 0.105 m mol) in dry dimethylformamide (1 ml) at -50 C under nitrogen was treated with mercuric acetate (0.033g, 0.105 m mol) in dimethylformamide (1 ml) followed by a saturated solution of ammonia in dimethylformamide (0.002g in 0.1 ml). The mixture was stirred for 1h during which time the temperature was allowed to rise to -10 C.
The mixture was then diluted with ethyl acetate (30 ml), washed with water (3 x 10 ml), brine (20 ml), dried (Mg SO4), and evaporated to give the title compound as a semi-solid material (0.044g, 98%); v,,,.
(CH2CI2) 1795, 1750 1650 cm-1; #[CDCl3 + (CD2)2CO] 7.42-6.85 (11H, m, aryl-H's and NH), 5.18 (4H, m, 3-H, 5-h and CH2Ph), 4.58(2H, s, OCH2), 3.33(2H, m, 2-H2) and 2.60 (2H, brs, NH2).
(e) Benzyl 6ss-phenoxyacetamido-6&alpha;-formamidopenam-3-carboxylate Benzyl 6ss-phenoxyacetamido-6&alpha;-aminopenam-3-carboxylate (0.042g, 0.103 m mol) in dry dichloromethane (5 ml) at OOC under nitrogen was treated with pyridine (0.08 g, 0.084 ml, 1.03 m mol) and then acetic formic anhydride (0.045 g, 0.52 m mol). The mixture was allowed to warm to room temperature and stirred for 3h. When it was washed with dilute hydrochloric acid, dilute sodium hydrogen carbonate, brine, dried (MgSO4) and evaporated.The crude product was chromatographed on silica gel 60 ( < 230 mesh ASTM) to give the title compound as a colourless gum (0.01g, 20%); vmax. (CH2CI2) 1795, 1745, 1690 and 1260 cm--1; â(CDCI3) 8.21(1K, d,J 1Hz, NH CHO), 7.89 (1H, d, J 1Hz NH CHO), 7.45-6.86 (11H, m, aryl; H's and NH), 5.48(1H, s, 5-H), 5.17 (2H, m, CH2Ph), 5.04 (1H, d, J 7Hz, 3-H), 4.52 (2H, m, PhOCH2), 3.27 (1H, d, J 11Hz, 2-H), and 2.84 (1H, dd, J11 and 7Hz, 2-H).
The benzyl group is removed by hydrogenolysis under conventional conditions to give 6ss-phenoxyace- tamido-60L-formamidopenam-3-carboxylic acid.
Example 22 7&alpha;-Formamido-7ss-[DL-2-phenoxycarbonyl-2-(thien-3-yl)acetamido]-3-methyl-1-oxadethia-ceph-3-em-4-car boxylic acid (a) t-Butyl 7&alpha;-methylthio-7ss-trichlorethoxycarbonylamino-3-methyl- 1-oxadethia-ceph-3-em-4-carboxylate Trichloroethylchloroformate (64mg; 0.3 mmol) in anhydreus methylene dichloride (1 ml) was added dropwise to a well stirred mixture of t-butyl 7ss-amino-7&alpha;-methylthio-3-methyl-1-oxadethia-ceph-3-em-4- carboxylate, toluene-p-sulphonic acid salt (129mg; 0.27 mmol) and pyridine (56mg; 0.7 mmol) in anhydrous methylene dichloride (5ml0 at 0 C.After 0.25h the reaction mixture was washed with dilute hydrochloric acid, dilute aqueous sodium hydrogen carbonate, brine dried (MgSO4), and evaporated.
Chromatography of the residue on silica gel afforded the title product (112mg; 93%); p,,,,, 3400, 1790, 1745, 1730, 1640 cm-l; # (CDCl3) 1.55 (9H,s), 1.99 (3H,s), 2.40 (3H,s), 4.29 (2H,s), 4.73 and 4.80 (2H, ABq, J 11 Hz), 4.93 (1H,s), 5.7 (1H, br s, exch.).
(b) t-Butyl 7&alpha;-amino-7ss-trichloroethoxycarbonylamino-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate t-Butyl 7a-methylthio-7P-trichloroethoxyca rbonylamino-3-methyl-1 -oxadethia-ceph-3-em-4-carboxylate (100mg; 0.225 mmol) in dry dimethylformamide (2ml) at -50 C under argon was treated with mercuric acetate (72mg; 0.225 mmol) in dimethylformamide [0.4ml). followed immediately by ammonia (6.5mg; 0.38mmol) in dry dimethylformamide (0.3ml). After stirring at -50 C for 0.75h, the reaction mixture was poured into ethyl acetate and washed well with each water and brine.The organic solution was dried over magnesium sulphate, evaporated and the residue chromatographed on silica gel to afford the title product (74mg; 79%); vmax (CHCI3) 3400, 1790, 1735, 1715, 1640cm-1; 3 (CDCl2) 1.55 (9H,s), 1.99 (3H,s), 2.2-2.9 (2H, br s, exch.), 4.33 (2H,s), 4.77 (2H,s), 4.89 (1H,s), 6.01 (1H, br s, exch.).
Also isolated from the reaction mixture was t-butyl 7ss-amino-7&alpha;-trichloroethoxycarbonylamino-3- methyl-l -oxadethia-ceph-3-em-4-ca rboxylate (6mg; 6%); vmax (CHCl3) 3400, 1790, 1740, 1718, 1640, cm; 3 (CDCI3) inter asia 1.52 (9H,s), 1.96 (3H,s), 1.9-2.6(2H,s, exch.), 4.37 (2H,s), 4,77 (2H,s), 5.13 (1H,s), 5.94 (1H,s, exch.).
(c) t-Butyl 7a-formamido-7(3-trichloroethoxycarbonylamino-3-methyl- i-oxadethla-ceph-3-em-4-carboxy- late t-Butyl 7cc-amino-7P-trich loroethoxycarbonyla mino-3-methyl-l -oxadethia-ceph-3-em-4-carboxylate (60mg; 0.145 mmol) was dissolved in dry methylene chloride (2ml) containing pyridine (115mg; 1.45 mmol), cooled to 0 C under argon and treated with acetic formic anhydride (64mg; 0.725 mmol). The reaction was stirred at 0 to 10 C for 1h., diluted with dry toluene and evaporated to dryness.The residue was redissolved in dry toluene and the solution evaporated; this was repeated twice.Chromatography of the residue on silica gel afforded the title product (63mg; 100%); vmax (CHCI2) 3400, 1795, 1740, 1700, 1690 sh cm -; 3 (CDCl3) 1.54 (9H,s), 2.01 and 2.03 (together 3H), 4.36 and 4.41 (together 2H), 4.74 and 4.78 (together 2H), 5.06 and 5.28 (together 1H), 6.21 and 6.45 (1H, exch.), 8.29 (d, J 1Hz) and 8.61 (d, J 11.6Hz) (together 1 HI.
(d) t-Butyl 7P-amino-701-formamido-3-methyl- 1-oxadethia-ceph-3-em-4-carboxylate t-Butyl 7a-fo rmam ido-7P-trich loroethoxyca rbonylamino-3-methyl-1 -oxadethia-ceph-3-em-4-carboxylate (33mg; 0.075 mmol) was dissolved in tetrahydrofuran (2ml) and potassium dihydrogen phosphate (1M; 0.4mi) and vigorously stirred with activated zinc (60mg; 0.92 mmol) at room temperature for 3h. The mixture was diluted with ethyl acetate, filtered through Kielseguhr, and the organic phase separated, washed with brine, dried (MgSO4), and evaporated.Chromatography of the residue on silica gel gave the title compound (14mg; 70%), which showed a 3:1 ratio of the two preferred conformations of the formamido group; x,,,, (EtOH) 272nm (E 6916); vmax (CHCl3) 3400, 3275, 1780, 1700, 1640 sh cm 3 (CDCI2) trans 1.56 (9H,s) 1.98 (3H,s) 1.9-2.5 (2H,s, exch.), 4.35 (2H,s), 5.14 (1H,s), 6.29 (1H, broad s, exch.), 8.24 (1H,d,J1.1Hz); cis 1.54 (9H,s), 1.9-2.5 (2H,s,exch.), 2.02(3H,s) 4.35 (2H,s), 4.87 (1H,s), 6.05 (1H,d,#11.6 Hz exch.), 8.42(1H,d,#11.6 Hz).
(e0 t-Butyl 7&alpha;-formamido-7ss-[DL-2-phenoxycarbonyl-2-(thien-3-yl)acetamido]-3-methyl-1-oxadethia-ceph- S-em-4-carboxylate DL-Phenoxycarbonyl-2-(thien-3-yl)acetic acid (27mg; 0.11 mmol) in an hydrous methylene chloride (1 ml) containing a catalytic amount of dimethylformamide, was treated with oxalyl chloride (21mg; 0.16 mmol). After stirring at room temperature for 1.5h., the solution was evaporated to dryness, treated with methylene chloride and re-evaporated. The process was repeated twice.The resulting acid chloride was taken up in dry methylene dichloride (0.5ml) and added dropwise with stirring to a mixture of t-butyl 7ss- amino-7&alpha;-formamido-3-methyl-1-oxadethia-ceph-3-em-4-carboxylate (20mg; 0.075 mmol) and pyridine (9mg; 0.11 mmol) methylene chloride (2ml) at -10 C. After 15 min. the reaction mixture was diluted with ethyl acetate, washed successively with dilute hydrochloric acid, dilute aqueous sodium hydrogencarbonate, brine, dried (MgSO4), and evaporated. Chromatography of the residue on silica gel gave the title compound (23mg, 57%); #max (EtOH) 263nm (# 13545), 314 nm (5545); vmax (CHCl3) 3400, 1790, 1740, 1720 sh., 1690 cm '; 3 (CDCI3) 1.52 (9H,s), 1.97 and 2.00 (together 3H,s), 4.15-4.40 (2H,m), 4.91 and 4.95 (together 1H,s), 5.21 and 5.23 (together 1H,s), 7.0-7.5 (10H,m), 7.80(-0.5H,s), 8.16, 8.19 and 8.52 (together 1H0, 8.22 (-0.5H,s).
The t-butyl protecting group is removed under conventional conditions to afford 7&alpha;-formamido-7ss-[DL- 2-phenoxycarbonyl-2-(thien-3-yl)acetamido]-3-methyl-1-oxadethia-ceph-3-em-4-carboxylic acid.

Claims (2)

1. A compound selected from the following; Ga-am ino-6p-phenoxyaceta m ido penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[2-(p-nitrobenzyloxycarbonyl-2-phenylacetamido]penicillanic acid, the benzyl ester thereof 6&alpha;-amino-6ss-[2-(p-nitrobenzyloxycarbonyl-2-(3-thienyl)acetamido penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl) acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;;-amino-6ss-[L-2-[(4-ethyl-2,3,-dioxopiperazin-1-yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl) acetamido] penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[D-2-(4-nitrobenxyloxycarbonylamino)-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[D-2-(4-nitrobenxyloxycarbonylamino)-2-(4-benzyloxycarbonyloxyphenyl)acetamido] penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[D-2-[(4-benzyloxycarbonyloxy) phenyl]-2-(2,2,2-trichloroethoxycarbonylamino)-acetamido] penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[DL-2-[4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino]-2-(thien-2-yl)-acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;;-amino-6ss-[DL-2-(4-nitrobenxyloxycarbonylamino)-2-(thien-2-yl)acetamido] penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-[(2R,3S)-3-benxyloxy-2-[(4-ethyl-2,3-dioxopiperazn-1-yl)carbonylamino]-butyramido] penicillanic acid, the benzyl ester thereof, 6&alpha;-amino-6ss-(2,2,2-trichloroethoxycarbonylamino)-penicillanic acid, the benzyl ester thereof, 7&alpha;-amino-7ss-(trichloroethoxycarbonylamino)cephalosporanic acid, the t-butyl ester thereof, 7&alpha;-amino-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1-yl) carbonylamino] 2-phenylacetamido]cephalosporanic acid, the t-butyl ester thereof, 7&alpha;;-amino-7ss-[D-2-[4-(benzyloxycarbony)phenyl-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino] acetamido]cephalosporanic acid, the t-butyl ester thereof, 6&alpha;-amino-6ss-phenoxyacetamido-penam-3-carboxylic acid, the benzyl ester thereof, 7O-am ino-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl) carbonylamino]-2-phenylaceamido]-3-methyl-1 -ox adethia-ceph-3-em-4-carbexylic acid, the t-butyl ester thereof, 7e-amino-7ss-trich loroethoxyca rbonylam ino-3-methyl-1 -oxadethia-ceph-3-em-4-ca rboxylic acid, the t-bu tyl ester thereof, 6&alpha;-amino-6ss(R-2-phenyl-2-sulphoacetamido penicillinic acid, triethylammonium salt, the benzyl ester thereof
2.A compound selected from the following; 6&alpha;-methylthio-6ss-[2-(p-nitrobenzyloxy-carbonyl)-2-phenyl-acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-methylthio-6ss-[-2-(p-nitrobenxyloxy-carbonyl)-2-(3-thienyl)acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-methylthio-6ss-[D-2-[(4-ethyl-2, 3 dioxopiperazin-1-yl) carbonylamino]-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, Ga-methylthio-GP-[D-2-[(4-ethyl-2, 3-dioxopiperazin-1 -yl) carbonylamino]-2-(4-benzyloxycarbonyloxyphenyl)-acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;;-methylthio-6ss-[L-2-4[(ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-(4-benzyloxycarbonyloxy- phenyl)acetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-methylthio-6ss-[D-2-(4-nitrobenzyloxycarbonylamino)-2-phenylacetamido]penicillanic acid, the benzyl ester thereof, 6&alpha;-(methylthio)-6ss-[DL-2-(4-nitrobenzyloxycarbonyl- amino)-2-(thien-2-yl)acetamide]penicillanic acid, the benzyl ester thereof, 6&alpha;-(methylthio)-6ss-[2R,3S)-3-benzyloxy-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-butyramido]- penicillanic acid, the benzyl ester thereof, 6&alpha;;-(methylthio-6ss-(2,2,2-trichloroethoxycarbonylamino-penicillanic acid, the benzyl ester thereof, 7a-methylthio-7(3-(trichlereethexycarbenylamine)cephalesporanic acid, the t-butyl ester thereof, 7(x-methylthio-7ss-[D-2-[(4-ethyl-2,3-dioxopiperazin-1 -yl)carbonylamino]-2-phenylacetam- ido]cephalosporanic acid, the t-butyl ester thereof, 7&alpha;-methylthio-7ss-[D-2-[4-benzyloxycarbonyloxy)-phenyl-2-[(4-ethyl-2,3-dioxopiperazin-1-yl)-carbonylam- ino)-acetamido]cephalosporanic acid, the t-butyl ester thereof, 6&alpha;;-(methylthio)-6ss-amino-penam-3-carboxylic acid, the benzyl ester thereof, 6ce-(methytthio)-6ss-phenoxyacetamido)-penam-3-carboxylic acid, the benzyl ester thereof, 7ss-(4-nitrobenzylideneamino)-7a-(methylthio)-3-methyl-1-oxadethia-ceph-3-em-4-carboxylic acid, the benzyl ester thereof, 7ss-amino-7(x-(methylthio)-3-methyl-1-oxadethia-ceph-3-em-4 carboxylic acid, toluene p-sulphonic acid salt, the benzyl ester thereof Ga-methylthio-GP-[D-2-[4-(benxyloxyca rbonyl oxy)phenyl]-2-2(2,2,2-trichl oroethoxycarbonylam- ino)acetamido] penicillanic acid, the benzyl ester thereof 6&alpha; ;-methylthio-6ss-[DL-2-[(4-ethyl-2, 3-dioxopiperazin-1 -yl )]-2-(thien-2-yl)acetamido] acetamido] penicillanic acid, the benzyl ester thereof, 7ss-[D-2-[(4-ethyl, 2, 3-dioxopiperazin-1-yl)carbonyi amino] -2-phenylacetamido-7&alpha;-methylthio-3-methyl-1-oxadethia-ceph-3-em-carboxylic acid, the t-butyl ester thereof, 7&alpha;-methylthio-7ss-trichloroethoxycarbonylamino-3-methyl-1-oxodethia-cepha-3-em-4 carboxylic acid, the t-butyl ester thereof, 6&alpha;-methylthio-6ss-(R-2-phenyl-2-sulphoacetamido) penicillanic acid,triethylammenium salt, the benzyl ester thereof,
GB08514519A 1981-07-25 1985-06-07 Intermediates for b-lactam antibiotics Expired GB2161803B (en)

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GB8209954 1982-04-03
GB8215007 1982-05-22
GB08221059A GB2107307B (en) 1981-07-25 1982-07-21 B-lactum antibacterial agents
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1348984A (en) * 1970-06-16 1974-03-27 Merck & Co Inc Antibiotics and processes for their production
US3962214A (en) * 1974-07-11 1976-06-08 Eli Lilly And Company Process for preparing amino substituted β-lactam antibiotics
GB1445446A (en) * 1972-12-06 1976-08-11 Squibb & Sons Inc Preparation of 7-substituted cephalosporins

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1348984A (en) * 1970-06-16 1974-03-27 Merck & Co Inc Antibiotics and processes for their production
GB1445446A (en) * 1972-12-06 1976-08-11 Squibb & Sons Inc Preparation of 7-substituted cephalosporins
US3962214A (en) * 1974-07-11 1976-06-08 Eli Lilly And Company Process for preparing amino substituted β-lactam antibiotics

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