DE2114541A1 - Cephalosporin antibiotics - Google Patents

Description

Cephalosporin
134/135/139/140

GLAXO LABORATORIES LIMITED, Greenford, Middlesex / Great Britain

Cephalosporin antibiotics

The present invention relates to improvements in antibiotics affect. More particularly, the present invention relates to a new group of carboxycephalosporin antibiotics. . .

The cephalosporin compounds, referred to in the present specification Reference is made, are generally referred to with regard to Cepham (cf. J.Am.Chem.Soc., 1962, 84, 3400). The term "cephem" denotes the basic cepham structure with a single double bond. If a dotted Line connecting the 2, 3 and 4 positions indicates that the connection is a Ceph-2-em or a Ceph-3-em connection can be.

It is known that ^ -4-carboxycephalosporin antibiotic compounds which are generally represented by the following formula:

1098A3 / 19B4

COOM

1 ζ

wherein R is a carboxylic acyl group and R is the

Is 3-substituent.

The new compounds according to the invention are characterized in that they have a substituent in the 3-position in which a carbonyl group is linked to the cephalosporin nucleus via two carbon atoms. The connections thus have the following structure

ι »-CH ₃ -CC-

in the 3 position. These compounds have activity against a variety of Gram-positive and Gram-negative Organisms.

According to one embodiment of the invention, the following compounds the general formula

q R ²

R NH-J ρ η,

J, —- N ^ J- CH ₅ -C-CO-R ⁴ ι

0 J 'COOH _R J

1 2 3

created, wherein R is a carboxylic acyl group, R and R, which are identical or different, hydrogen, lower-alkyl, phenyl, substituted phenyl, lower-alkoxycarbonyl, mono- or diaryl-lower-alkoxycarbonyl, lower-alkyl carbonyl, aryl-lower-alkyl or C _c - or C ^ cycloalkyl and

4 do

R is hydrogen, lower alkyl, phenyl, substituted phenyl,

Aryl-lower alkyl or C ₅ - or Cg -cycloalkyl, and their non-toxic derivatives, e.g. base salts (if available )

109843/1954

_ 3 -

and acid addition salts (if available) and compounds, obtained by reacting an amino group and / or acylamido group (s) the 7-side chain with a ketone, e.g. acetone or methyl ethyl ketone, or an aldehyde.

The term "non-toxic" as applied to compounds of the present invention Invention used v / ird, denotes those derivatives which, at the dosage in which they are administered, are physiologically compatible.

Such derivatives include salts and esters.

Since optically active centers in the substituents in the 3- and / or 7 positions, includes the present Invention also the various diastereoisomers as well as their mixtures.

The compounds according to the invention have antibacterial properties Activity against a wide variety of gram-positive and gram-negative Organisms and are therefore of value in human and veterinary medicine. You can also help in the manufacture of others 3-substituted cephalosporin compounds are used will.

The terms "lower alkyl" and "lower alkoxy" as used herein indicate that the Groups contain 1 to 8 carbon atoms.

2 3 4

If the groups R, R or R represent lower alkyl groups, so they preferably mean the methyl or ethyl group.

2 3 4 If one of the substituents R, R or R is a substituted Means phenyl group, the substituent of the phenyl group can be one or more halogen atoms (e.g. chlorine or bromine), lower Alkyl groups, lower alkoxy groups, nitro groups, amino groups or lower alkylamino groups.

1 0 BB L 3/1 9 B U

_ 4 -

The group R in the above formula can represent a variety of acyl groups containing 1 to 20 carbon atoms can*. Special acyl groups are listed below, but without representing a limitation:

i) R ^U CH ₂ CO-, where R ^{u is} an aryl (carbocyclic or heterocyclic), cycloalkyl, substituted aryl, substituted cycloalkyl, cyclohexadienyl group or a non-aromatic heterocyclic or mesionic group and η is an integer represents from 1 to 4. Examples of this group include the phenylacetyl, substituted phenylacetyl, e.g., fluorophenylacetyl, nitrophenylacetyl, aminophenylacetyl, acetoxyphenylacetyl, methoxyphenylacetyl, methylphenylacetyl or hydroxyphenylacetyl groups; N, N-bis (2-chloroethyl) aminophenylpropionyl group, the thien-2- and -3-ylacetyl group, the 4-isoxazolyl and substituted 4-isoxazolylacetyl group; the pyridylacetyl group; the tetrazolylacetyl group or a sydnonacetyl group. The substituted 4-isoxazolyl group can be a 3-aryl-5-methyl-isoxazol-4-yl group, the aryl group being a phenyl or halophenyl group, for example a chlorine or bromophenyl group. An acyl group of this type is the So-chlorophenyl-S-methylisoxazol-4-yl-acetyl group.

ii) C Hp .CO-, where η is an integer from 1 to 7. The alkyl group can be straight or branched and, if desired, through an oxygen or sulfur atom interrupted and e.g. by a cyano group, a carboxy group, an alkoxycarbonyl group, a hydroxyl group or a carboxycarbonyl group (-CO-COOH) substituted be. Examples of such groups include the cyanoacetyl, hexanoyl, heptanoyl, octanoyl and butyl thioacetyl group. -

iii) ^c _n ^H 2n_i ^C0 ~ * ^where i ^{nn is} an integer from 2 to 7. The alkenyl group can be straight-chain or branched

109843/19 54

21H541

a
and if desired by / oxygen or sulfur atom

'be interrupted. An example of such a group is the allylthioacetyl group.

iv) R ^V

R ^U OC-CO-, where R ^{u has} the meaning given under i)

R ^w

sits and can also represent the benzyl group, and R ^v and R ^w , which are identical or different, each denote hydrogen, the phenyl, benzyl, phenethyl or lower alkyl group. Examples of such groups include the phenoxyacetyl, 2-phenoxy-2-phenyl- 'acetyl, benzyloxyacetyl, 2-phenoxypropionyl, 2-phenoxybutyryl, methylthiophenoxyacetyl groups.

v) R ^V ι

R ^U SC-CO-, where R ^{u has} the meaning given under i)

R ^w

sits and can also mean the benzyl group and

ν w
R and R have the meaning given under iv).

Examples of such groups include the S-phenylthioacetyl, S-chlorophenylthioacetyl, S-fluorophenylthioacetyl, Pyridylthioacetyl and S-benzylthioacetyl groups.

vi) R ^U Z (CH ₂ ) _m CO-, where R ^{u has} the meaning given under i) and can also mean the benzyl group and Z is an oxygen or sulfur atom and m is an integer from 2 to 5. An example of such a group is the S-benzylthiopropionyl group.

vii) R ^U CO-, where R ^{u has} the meaning given under i). Examples of such groups include the benzoyl, substituted benzoyl (eg aminobenzoyl), 4-isoxazolyl carbonyl and substituted 1-Isoxazolylcarbonyl-, cyclo pentancarbonyl-, Sydnoncarbonyl-, naphthoyl and substituted naphthoyl (eg 2- ithoxynaphthoyl-) , quinoxalinyl-

109843/1954

21H541

carbonyl and substituted quinoxalinylcarbonyl groups (e.g. the 3-carboxy-2-quinoxalinylcarbonyl group). Other possible substituents for the benzoyl group include

Alkyl, alkoxy, phenyl, substituted by carboxy

f. phenyl, alkylamido, cycloalkylamido, allylamido,

Phenyl (lower) alkylamido, morpholinocarbonyl, pyrrolidinocarbonyl, piperidonocarbonyl, tetrahydropyridine, furfurylamido or N-alkyl-N-anilino groups or their derivatives, and such substituents can be in the 2- or 2- and 6 Positions are available. Examples of such substituted benzoyl groups are the 2,6-dimethoxybenzoyl, 2-methylamidobenzoyl and 2-carboxybenzoyl groups. If the group R ^{u represents} a substituted 4-isoxazolyl group, the substituents can have the meanings given under i). Examples of such isoxazolyl groups are the S-phenyl-S-methyl-isoxazol-4-yl-carbonyl group, the So-chlorophenyl-S-methylisoxazol-4-yl-carbonyl group and the 3- (2,6-dichlorophenyD-5-methyl -isoxazol-4-yl-carbonyl group.

viii) R ^U -CH-CO-, where R ^{u is} the meaning given under i)

and X is an amino group, a substituted amino group (e.g. the acylamido group or a group obtained was made by reacting the α-aminoacylamido group of the 7-side chain with an aldehyde or ketone, e.g. acetone, Methyl ethyl ketone or ethyl acetoacetate, the hydroxy, carboxy, esterified carboxy, azido, triazolyl, tetrazolyl-j Cyano-, halogen-, acyloxy-τ (e.g. formyloxy- or low alkanoyloxy) or etherified hydroxyl group. Examples of such acyl groups are the a-aminophenylacetyl and the α-carboxyphenylacetyl group.

ix) R ^x

. R ^Y -C-CO-, where R ^x , R ^y and R ^z , which are identical or different

R ^z
can be different, each with a lower alkyl, phenyl

109843 / 18S4

or substituted phenyl group or R represents a hydrogen atom. An example of such a The acyl group is the triphenylmethylcarbonyl group.

x) R ^U -NH-CO-, where R ^{u has} the meaning given under i) and additionally hydrogen, the lower alkyl or halogen-substituted lower alkyl group. An example of such a group is C1 (CH ₂ ) ₂ NHCO.

(CH ₉ ) ^ C-CO-, in which X is given under viii)

Has meaning and η is an integer from 1 to 4. An example of such an acyl group is the 1-aminocyclohexanecarbonyl group.

xii) Aminoacyl, for example R ^W CH (NH ₃ ) - (CH ₂ J _n CO-, where η is an integer from 1 to 10 or NH ₂ -C _n H _2n2 ^, where m is 0 or an integer of 1 to 10 and η are 0, 1 or 2, R ^{W is} a hydrogen atom or an alkyl, aralkyl or carboxy group or a group as defined above under R ^u , and Ar is an arylene group, for example p-p-phenylene or 1 , 4-naphthylene group. Examples of such groups are described in British Patent 1,054,806. One group of this type is the p-aminophenylacetyl group. Other acyl groups of this type include those derived from naturally occurring amino acids, e.g. -Aminoadipoyl group, or derivatives thereof, for example the N-benzoyl-5-aminoadipoyl group.

xiii) Substituted glyoxylyl groups of the formula R ^Y -CO-CO-, in which R ^y denotes an aliphatic, araliphatic or aromatic group, for example a thienyl group, a phenyl group or a mono-, di- or trisubstituted group

109843/1954

Denotes a phenyl group, the substituents being e.g. or several halogen atoms (F, Cl, Br or J), methoxy groups, methyl groups or amino groups or a fused-on Are benzene ring. The α-carbonyl derivatives of the above-mentioned substituted ones are also included in this group Glyoxylyl groups included.

xiv) formyl groups. .

If the compounds of general formula I as intermediates Are used are important meanings of the group; R the following:

'' \ hydrogenoxy-

xv) Hydrocarbon oxycarbonyl and substituted carbon \ groups (in which the 7-amino group is part of a urethane), e.g. lower alkoxycarbonyl groups (such as the methoxycarbonyl, Ethoxycarbonyl and tert-butoxycarbonyl groups); Halo-lower alkoxycarbonyl groups such as the 2,2,2-trichloroethoxycarbonyl group; Aralkoxycarbonyl groups, such as. B. the benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, Diphenylmethoxycarbonyl and 4-nitrobenzyloxycarbonyl groups; and cycloalkoxycarbonyl groups such as the adamantyloxycarbonyl group.

xvi) haloformyl groups, e.g. the chloroformyl group.

An important group of compounds according to the invention are the compounds of the general formula

R ^D -CHCONHn [

Il JN ^ y ^ - CH ₂ -C-CO-R * II

where R is an aromatic group, for example the phenyl group or phenyl group substituted with halogen atoms, hydroxy, lower alkyl, nitro, amino, lower alkanoyl, lower alkoxy or lower alkyl! mercapto groups} the naphthyl group or a hetero-

109843/1964

cyclic group (especially a 5- or 6-membered heterocyclic group which has at least one heteroatom such as S, N or 0, for example the thien-2-yl or thien-3-y! group) ;; X ¹ the amino, substituted amino, e.g. the acylamido

group, the hydroxyl group, the formyloxy group, low alkano-

2 3 4

yloxy groups, or hydrogen and R, R and R are the Have the meanings given above, as well as their non-toxic derivatives.

The compounds of general formula II in which the corresponding acid R ⁵ CH (X ¹ ) COOH (in which R ⁵ denotes the phenyl or substituted phenyl group and X denotes the amino or hydroxyl group) belongs to the D series are of particular interest Interest.

An important subgroup of compounds of formula II are the compounds of the general formula

III

wherein R _1, R and R have the meanings given above, as well as their non-toxic derivatives.

An example of a compound of the general formula III is 7β- (2-amino-2-phenylacetamido) -3- (2-methyl-3-oxopentyl) -cepJt ** 3-em-4-carboxylic acid the formula

CH-CONH L

₂ CH ₃ IV

109843/1964

21U541

7ß- (D-2-Amino-2-phenylacetamido) -3- (2-methyl-3-oxo-pentyl) -ceph-3-em-4-carboxylic acid is a broad spectrum antibiotic that is active against Gram- positive and gram negative organisms as shown by in vitro tests to be effective. Animal studies have shown that this compound is essentially resistant to degradation in vivo.

Another subgroup of important compounds of the formula II are compounds of the general formula

R ⁵ -CHCONH-j f l

i r ^N > _N ^ ~ CH ₂ CH ₂ -COCH ₃

COOH

in which R and X have the meanings given above, as well as their non-toxic derivatives.

An example of a compound of the formula V is 7β- (2-amino-2-phenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid of formula

CH ₂ -CH ₂ -CO-CH ₃ VI

The 7ß- (D-2-amino-2-phenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid is an antibiotic with a broad spectrum of action, which is effective against gram-positive and gram-negative organisms, as shown in in vitro studies

The compounds of the formulas IV and VI show activity against Staphylococcus aureus, Escherichia coll, Proteus mirabilis, Klebsiella aerogenes, Hafnia _t Aerobacter cloacae and S. typhimurium. . ■

109843/1954

21U541

Another example of a compound of the formula V is 3- (3-oxobutyl) -7β- (2-thienylacetaraido) -ceph-3-em-4-carboxylic acid the formula

CH ₂ CONH

-N _n ^ jJ CHpCHpCOCH,

VII

CO ₂ H

and their non-toxic derivatives. This compound is a broad spectrum antibiotic that works against a number of Gram-positive and Gram-negative organisms is effective, vvj, e was found from in vitro studies.

An important series of "connections" with those of the formulas III, IV and VI are related are those in the 7-position of the group of the formula

ti

-CH ' N

HN

where R and R , which may be identical or different, are lower alkyl groups (C ^ to C.), in particular methyl or ethyl groups, or isomeric forms of the groups and their salts.

Salts which can be formed from the compounds according to the invention include a) inorganic base salts, such as alkali metal salts, for example sodium and potassium salts, alkaline earth metal salts, for example calcium salt, and salts of organic bases, for example procaine and dibenzylethylenediamine salts, and b) acid addition salts, for example with hydrochloric acid, hydrobromic acid . ,,

109843/1954

21U541

acid, sulfuric acid, nitric acid, phosphoric acid, toluene-sulfonic acid and methanesulfonic acid. The salts can also be in the form of resinates, e.g. with a polystyrene resin, the amino, quaternary amino or sulfonic acid groups or with a resin having carboxyl groups such as a polyacrylic acid resin. The resin can, if desired be crosslinked, i.e. it can be a copolymer of styrene and divinylbenzene that contains the corresponding groups.

According to another object of the present invention is a process for the preparation of a compound of the formula

R ² CH - C-CO-R

COOH R

1 2

created, wherein R is a carboxylic acyl group, R and

R, which can be identical or different, hydrogen, lower alkyl, phenyl, substituted phenyl, lower alkoxycarbonyl, lower alkylcarbonyl, mono- or diaryl-lower alkoxycarbonyl, aryl-lower alkyl or C ₁ - -

4 or Cg-cycloalkyl groups and R is hydrogen, a represents lower alkyl, phenyl, substituted phenyl, aryl-lower alkyl or Cc or Cg cycloalkyl group, as well as their non-toxic derivatives, which are characterized is that you A) a compound of formula

, 2

VIII

wherein R is hydrogen or a carboxyl blocking group,

2 3 4? > S or> S - »0 (α- or ß-) mean and R, R and R have the meanings given above, with an acylie

109843/1954

ORIGINAL INSPECTED

-υ-

21H541

agent, that of the acid

R ¹ OH

1
corresponds to, wherein R has the meaning given above, acylated or

B) a compound of the formula

IX

COOR

Ί 8
wherein R, R and Z have the meanings given above and X is chlorine, bromine or iodine, with an enamine of the formula

C-R

2 3 4
wherein R, R and R have the meanings given above

ο
and the R groups, which can be identical or different, represent either lower alkyl groups or a divalent aliphatic group, preferably a C ₄ or C ^ group, which together with the adjacent nitrogen atom forms a heterocyclic ring, and the resulting Bringing reaction mixture with an aqueous medium, for example water, or

C) a compound of the formula

^ix

COOR

1 8 wherein R , R , Z and X have the meanings given above, with an anion which is replaced by a carbanion of the formula

109843/1954

21H541

0c - CO - R ⁴

2 3 4
ivorin R, R and R have the meanings given above, can be represented, reacted, after which, if necessary, the following reactions

D) are carried out:

i) decarboxylation of a compound in which R is a lower alkoxycarbonyl or mono- or diaryl-lower alkoxycarbonyl group, means,

ii) conversion of a ^ -isomer into the desired ^ -isomer,

üi) separation of the amino or carboxyl protective groups and

iv) reduction of a compound in which Z is ^ S - ^ 0 for Formation of the desired compound, in which Z -.> S means.

For the sake of simplicity, the group of the formula

^1νη -γ-Γ)

COOR ⁸ hereinafter referred to as Q-.

The present invention will now be described in more detail with reference to reactions A, B, C and D above.

A. Acylation

The acylation can be carried out with any suitable acylating agent be carried out, for example with an acid halide (e.g. the acid chloride or acid bromide), an anhydride or mixed Anhydride, e.g. with pivalic acid, or with a haloformate, e.g., a lower alkyl haloformate, or an active ester or azide; alternatively, the acid itself can be used together with an esterifying agent, e.g.

108843 / 1ΘΒ4

21 USA 1

-. 15 -

Carbonyldiimidazole, or a carbodiimide, such as N, N'-diethylcarbodiimide, Ν, Ν'-dipropylcarbodiimide or N, N'-diisopropylcarbodiimide or, preferably, NjN'-dicyclohexylcarbodiimide.

The cephalosporin compound can be used as a compound having a free 4-C00H group or can be in the 4-position have a blocking group, for example an ester with an alcohol or phenol, which can be obtained, for example, by hydrolysis or reduction can be split off at a later reaction stage, or can be used as an acid addition salt of the free 4-COOH compound or their esters e.g. with nitric acid or a hydrocarbon sulfonic acid are used, examples of hydrocarbon sulfonic acids include alkylbenzenesulfonic acids, e.g. p-toluenesulfonic acid, and lower alkanesulfonic acids, e.g. Methanesulfonic acid.

The group that protects the 4-carboxyl group can with a Alcohol (aliphatic or araliphatic), a phenol, silanol, stannanol or an acid are formed, which in can easily be split off at a later reaction stage.

Suitable esters therefore include compounds which, as a 4-ester group, are a non-exhaustive group of the following Compilation included:

i) -COOCR ³ RR ^c , in which at least one group R ^a , R ^b and R ^{c is} an electron donor, for example the p-methoxyphenyl, 2,4,6-trimethylphenyl, 9-anthryl, methoxy, acetoxy , Tetrahydrofur-2-yl, tetrahydropyran-2-yl or fur-2-yl group. The remaining groups R ^a , R and R ^c can be hydrogen or organic substituents. Suitable ester groups of this type include the p-methoxybenzyloxycarbonyl and 2 ^ 4,6-trimethylbenzyloxycarbonyl groups.

ii) -COOCR ^a RR ^c , in which at least one of the groups R ^a , R and R ^{c is} an electron-withdrawing group, for example a benzoyl, p-nitrophenyl, 4-pyridyl, trichloromethyl,

109843 / 1-964

Tribromomethyl, iodomethyl, cyanomethyl, ethoxycarbonylmethyl, Arylsulfonylmethyl-, 2-dimethylsulfoniumethylo-nitrophenyl- or cyano group. The remaining groups R, R and R can be hydrogen or organic substituents be. Suitable esters of this type include the benzoylmethoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl-, 2,2,2-trichloroethoxycarbonyl- and 2,2,2-tribromoethoxycarbony1 group.

iii) -COOCR ³ RR ⁰ , in which at least two of the groups R ^a , R

and R ^{c is} a hydrocarbon group such as an alkyl group, for example the methyl or ethyl group, or an aryl group, for example the phenyl group, and the remaining group R ^a , R and R ^c , if any, are hydrogen. Suitable esters of this type include the tertiary butyloxycarbonyl group, the tertiary amyloxycarbonyl group, the phenylmethoxycarbonyl group and the triphenylmethoxycarbonyl group.

iv) -COOR, where R is an adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl or tetrahydropyran-2-yl group mean.

Silyl esters can suitably be made from a halosilane or a silazane of the formula

R ⁴ ₃ SiX R ⁴ SiX R ⁴ SiNR ⁴ R ⁴ SiNHSiR ⁴ R ⁴ SiNHCOR ⁴

₃ SiX, R ₂ SiX ₂ , R ⁴ ₃ Si-NR ⁴ ₂ , R ⁴ ₃ Si-NH-SiR ⁴ ₃ , R ⁴ ₃ Si-NH-COR ⁴ , R ⁴ ₃ Si-NH-CO-NH-SiR ⁴ ₃ , R ⁴ NH-CO-NR ⁴ -SiR ⁴ ₃ or R ⁴ C (OSiR ⁴ ₃ ) = NSiR ⁴ ₃ , in which X is a halogen and the various groups R, which may be identical or different, hydrogen atoms or alkyl groups, for example methyl, ethyl, n-propyl and isopropyl groups, aryl groups, for example the phenyl group, * or aralkyl groups, for example the benzyl group.

Preferred silanol derivatives are silyl chlorides such as trimethylchlorosilane and dimethyldichlorosilane.

109843/1964

21U541

The carboxyl groups can be regenerated from the ester by conventional methods; e.g. the acid and base catalyzed Hydrolysis (especially for the silyl and stannyl esters) generally applicable as well as the enzyme-catalyzed ones Hydrolyses. However, aqueous mixtures can be poor solvents for these compounds, and they can be isomerizations, Rearrangements, side reactions and general destruction cause special processes to be desirable could be. Five suitable deesterification methods are as follows:

i) Reactions with Lewis Acids: Suitable Lewis acids for reacting with the esters include trifluoroacetic acid, Formic acid, hydrochloric acid in acetic acid, zinc bromide in benzene and aqueous solutions or suspensions of mercury-II compounds. The reaction with the Lewis acid can be increased by adding a nucleophilic reagent such as anisole.

ii) Reduction: Suitable systems for carrying out the reduction are zinc / acetic acid, zinc / formic acid, zinc / lower Alcohol, zinc / pyridine, palladium on charcoal and hydrogen, electrolysis and sodium and liquid ammonia.

iii) attack by nucleophilic agents: .Suitable nucleophilic Agents are those that are a nuc ~ eophilic oxygen or Contain sulfur atom, e.g. alcohols, mercaptans, thiocyanates and water.

iv) Oxidative processes such as those involving the use of hydrogen peroxide and acetic acid .

v) irradiation.

The acylation can be carried out in an aqueous medium with an acid halide, for example in an aqueous solution of one with water

1Q98A3 / 1954

21U541 -

ir - 18 -

miscible ketones, such as acetone, or in an aqueous solution of Tetrahydrofuran, preferably also in the presence of an acid-binding agent, can be carried out. Suitable acid-binding Agents include tertiary amines such as triethylamine, dimethylformamide, Dimethylaniline; inorganic bases such as calcium carbonate or sodium bicarbonate, and oxiranes, the hydrogen halide bind, which is released during the acylation reaction will. The oxirane is preferably a lower 1,2-alkylene oxide, e.g. ethylene oxide or propylene oxide.

The p "value in the acylation is during the reaction, the at temperatures from -30 C to +80 C, preferably from 0 to 25 ° C, is maintained at a value of 5 to 7. The acylation can also be carried out in an organic solvent, like ethyl acetate, e.g. under reflux *

Alternatively, the acylation with an acid halide or mixed anhydride under essentially anhydrous conditions in the liquid phase in an inert Lewis base (preferably a base that has a tertiary nitrogen atom) with a dielectric constant above 15 and preferably above 30 and which has a hydrogen halide acceptor contains, are effected. The reaction can be carried out with the free 4-COOH compound or an ester thereof or an acid addition salt, e.g. the hydrogen p-toluenesulfonate of the 4-COOH compound or the ester thereof. An acid addition salt of the above-mentioned ester can be used among those mentioned Use anhydrous conditions, since this derivative is in high yield from the N-deacylation stage described below can be obtained. After that, if desired the ester group can be separated. The acid halide is preferably the chloride or bromide.

Another alternative method of performing the acylation the use of an α-aminoarylacetyl chloride, the free amino group of which is preferred as the acid addition salt the hydrochloride is protected as an acylating agent.

109843/1964

21U5A1

According to a preferred embodiment of the process according to the invention, a process for the preparation of compounds the formula

q create,

2 3 4 5

wherein R, R, R and R have the meanings given above, which is characterized in that one compound the formula

/ S _n R ²

CH - C-COR ⁴ * ■ ι

3 CO ₂ H

N _n Y-

or an ester, a salt or a salt ester thereof with a compound of the formula

R ^ -CH-CO-Cl

NH ₂ -HCl reacts in the presence of an acid-binding agent.

Procedures for carrying out the acylation under essentially anhydrous conditions are detailed in the British patent No. 1,104,937.

In general, acylation procedures known in peptide chemistry can be used. For example, the 7-amino compound (VIII), for example in situ, can be converted into the analogous 7-isocyanate compound
will.

g compound converted with the acid R OH prior to acylation

109843/1954

21U5U

B. Reaction with enamines

As indicated above, the compounds of the formula I can be prepared by reacting a compound of the formula

Q - CH ₂ X IX

wherein Q and X have the meanings given above, with an enamine of the formula

9 9

2
R ^

CR ⁴ X

2 3 4 9

wherein R, R, R and R have the meanings given above, and the resulting reaction mixture with a aqueous medium, suitably an aqueous acidic medium, which also destroys the excess enamine, brings into contact.

The reactions of compounds of the formula IX with the enamines run easily when X = J.

The 3-halomethylcephalosporins IX can be prepared by halogenation of a 7β-acylamido-3-methylceph-3-em-4-carboxylic acid ester-β-oxide, if desired followed by the reduction of the β- oxide group, as described in Belgian patent specification No. 755 256, getting produced. The 3-halomethylceph-3-em compounds can also be prepared by the process described in Belgian Patent Specification No. 719,711. The corresponding Ceph-2-em compounds can be prepared by the _{method described in published Dutch patent application No. e} 69O2O13 by reacting a Ceph-2-egg-3-methyl = compound with N-bromosuccinimide to form the Ceph-2-em-3 -bromomethyl compound can be produced.

109843/1954

9 The enamine can react with an amine R p ^{NH m} $ -t a

Carbonyl compound of the formula

/ CH - CO - R ⁴ XI

R ³

2 3 4
wherein R, R and R have the meanings given above,

getting produced.

The reaction of the 3-Halogenmethylcephalosporinverbindung with the enamine can be conveniently carried out by adding to +100 ⁰ C to +90 ⁰ C, holding the reactants in solution at a temperature of, for example -80, preferably -20 °. The reactions are normally complete at 20 ^° C. in the course of 15 minutes. The reaction is advantageously effected using from 1 molar equivalent to 5 molar equivalents of the enamine. The excess enamine can be quickly destroyed by acid after the initial reaction is complete.

The reaction media used can be low alkylnitriles, for example acetonitrile or propionitrile, halogenated hydrocarbons, for example methylene chloride, carbon tetrachloride, chloroform, ethylene dichloride or perchlorethylene; lower nitroalkanes, for example nitromethane; nitroaromatic compounds, for example nitrobenzene; cyclic ethers, for example dioxane or tetrahydrofuran; aromatic hydrocarbons, for example benzene or toluene; Amides of the general formula R ^C -CO-NR -R ^e , in which R ^{c is} a hydrogen atom or an alkyl group having 1 to 5 carbon atoms

de
and R and R, which may be the same or different, each represent a hydrogen atom or an alkyl group having 1 to 5 carbon atoms or, alternatively, together represent a divalent aliphatic group which together with the adjacent nitrogen atom form a heterocyclic ring. Examples of amides of this type are N, N-dimethylformamide, N ₁ N-diethylformamide and Ν, Ν-dimethylacetamide.

109843/1954

21H541

The course of the reaction can be monitored by thin layer chromatography and by spectroscopic studies of the residue bound in the 3-position, e.g. when a new carbonyl frequency occurs in the infrared spectrum.

Since two reaction stages are run through, there is a certain one Trying out is necessary to ensure that the first reaction is complete before using water or an aqueous medium is admitted. In general, however, this is not a problem as the initial reaction is fairly quick.

The reaction product can be obtained from the reaction mixture, which contains, for example, unchanged cephalosporin and other substances, through a variety of processes including crystallization, ionophoresis, countercurrent separation, paper chromatography, Thin layer chromatography or, for example, chromatography over ion exchange resins or silica gel.

C. Reactions with carbanions

As already indicated above, the compounds of the formula I can be prepared by adding a compound of the formula

Q - CH ₂ X IX

wherein Q and X have the meanings given above, in particular in the form of the free carboxylic acid, with an anion or an equivalent nucleophile, which acts as the carbanion of

formula

«2

C-CO-R ⁴

2 3 4

can be represented in which R, R and R are those given above Have meanings.

109843/1954

21H5A1

The required carbanions can be obtained from the conjugate acid of the formula

CH-CO-R ⁴ • XI

can be obtained by treatment with a base. The carbanion can in situ in the process according to the invention Reaction of the conjugate acid with a base, preferably stronger than the carbanion, are formed. Suitable bases include alkali metal and alkaline earth metal hydroxides, carbonates and hydrogen carbonates, e.g., sodium hydroxide or Sodium bicarbonate. Other bases that can be used to form the carbanions include the conjugated Bases of dimethyl sulfoxide, dimethylacetamide, dimethylformamide and hexamethylphosphoramide; tertiary nitrogen bases, e.g. pyridine or trialkylamines such as triethylamine; the sodium or Lithium derivatives of hexamethyldisilazane; Alkali metal hydrides, e.g., sodium hydride; Alkali metal alcoholates, e.g., tert-butylate; Sodium triphenyl methylate; Alkylene oxides, e.g. Xthylene oxide or propylene oxide, with a halide ion. can be enhanced, or fluoride ion in an aprotic Solvent. Enol derivatives containing thallium-I can can also be used and these can be considered a source of the desired carbanions or equivalent nucleophiles will.

Although the carbanion can be made with 1 equivalent of the base, it is preferred to use about 2 equivalents when R = H since, unless the group at the 4-position of the cephalosporin starting material is present as -COp ", 1 Equivalent of the base is required to form the group -CO ₂ "* and thus a second equivalent is required to produce the carbanion. It is not desirable anymore

O-

to be used as about 3 equivalents of the base when R ■ denotes H, since anions are then present which can react instead of the carbanion in the process according to the invention.

109843/1954

The reaction may be carried out by mixing the ingredients, for example, to +35 ⁰ C, stirred at a temperature of -80 to +80 C, preferably -10 °. If the reaction is carried out at a temperature at which one or more of the reactants will volatilize, a closed system can be used. The reaction can be carried out in an inert or relatively inert solvent, for example a halogenated hydrocarbon, for example methylene chloride; a hydrocarbon, for example benzene or η-hexane; an acyclic or cyclic ether, for example diethyl ether, tetrahydrofuran or dioxane; in dimethyl sulfoxide; an amide, for example dimethylformamide or dimethylacetamide or hexamethylphosphoramide, can be carried out. The course of the reaction can be followed by thin-layer chromatography or paper chromatography and by UV spectrography (in general the X _{m = v} shift to higher wavelengths because the reaction forms chromophoric groups).

Protection of the amino groups

If the 7β-acylamido group contains an amino group, it is necessary to protect this group during the various stages of the reaction. The protecting group is suitably one which can be removed by hydrolysis without affecting the rest of the molecule, especially the lactam and 7β-amido bonds. The amine protecting group and the esterifying group in the 4-COOH position can be removed by the same reagent. One advantageous method is to split off both groups in the last stage of the sequence. Protected amine groups include urethane, arylmethylamino, for example trityamino, arylmethyleneamino, sulfenylamino or enamine groups. Such groups can generally be replaced by one or more reagents such as dilute mineral acids, for example dilute hydrochloric acid, concentrated organic acids, for example concentrated acetic acid, trifluoroacetic acid, and liquid. Hydrogen bromide, be separated at a very low temperature, for example -80 ° C. A suitable protective group is the tertiary ^! Butoxycarbonyl group, which can easily be formed by hydrolysis with dilute '

109843/1954

21U541

ter mineral acid, e.g. dilute hydrochloric acid, or preferably with a strong acid, e.g. B. formic acid,
Trifluoroacetic acid or liquid HF, for example at a temperature of 0 to 40 ° C, preferably at room temperature (15 to
25 ° C), can be split off. Another suitable protecting group is the 2,2,2-trichloroethoxycarbonyl group which can be cleaved by an agent such as zinc / acetic acid, zinc / formic acid, zinc / lower alcohols or zinc / pyridine. The NHp group can also be _{protected as an NH 3} ⁺ group using the amino acid halide as its hydrohalide under conditions in which the amino group remains protonated.

Typical protective groups and processes for their removal are summarized in the following table.

109843/1954

Tabel

ο co 00 4P *

co

Art example Common name and analogs etc. I'm leaving
procedure Urethane HACOCH ₂ Ph
0 Benaoyloxycarbonyl, p-methoxy HBr / AcOH (smooth)
CF ₃ COOH (smooth)
flow.HBr at -8O ° C Urethane HNCOC (CH-.),
Q tert-butoxycarbonyl dilute acid (HCl)
CP ₃ COOH (smooth) Urethane HNCOCHPh ₀
ti C
0 Diphenylmethoxycarbonyl CF ₃ COOH (smooth)
diluted HCl etc. Urethane HNCO- (1-adamantyl) 1-adamantyloxycarbonyl diluted HCl Arylmetlv fl HNCPh ₃ Trityl AcOH + H ₂ O
diluted HCl Sulfenyl NO ₂
HN-S- ^ o-nitrophenylsuIfenyl,
p-nitro diluted HCl
NaJ or
Na ₃ S ₃ O ₃
p _H 2 to 4

cn I

cn

Table (port setting)

ο to OO

co cn

Art example Common name and analogs etc. Usual parting
tion procedure Enatnin i ν
° ** _C _> ^H
I.
R. β-dicarbonyl
ReOEt ethyl acetoacetate
R »CH \ acetylacetone
R * Ph benzoylacetone
R »0Me methyl acetoacetate
R »C _? H ₅ propionyl acetone
and many other ß-diketones acid labile in under
to varying degrees
diluted AcOH or
HCl etc. Aryl
methylene , N-CH
HO-tp ^ Anil (similar to ß-dicarbonyl)
from salicylaldehyde
5-chlorosalicylaldehyde
3,5-dichlorosalicylaldehyde
2-hydroxy-1-naphthaldehyde
3-hydroxy-pyridine-4-aldehyde diluted HCl
Formic acid Omnium NH ₃ ⁺ base Urethane HN-CO-OCH ₂ CCl ₃ ß, ß, ß-Trichloräthoxycarbonyl Reducing agents,
e.g. *. Zn / acetic acid

-4 I.

<n

N-deacylation

The product of stages B or C can lead to the corresponding 7β-amino compound of the formula

I COOH

2 3 4

wherein R, R, R and Z have the meanings given above, or a derivative thereof (e.g. the ester, the salt or the salt of an ester) N-deacylated, and the 7β-amino compound can then with an acylating agent that the acid

R ¹ OH

corresponds, in which R has the meaning given above, are acylated. Acid addition salts, e.g. with nitric acid or a hydrocarbyl sulfonic acid, can with the free 4-COOH compound or the ester thereof. Examples of hydrocarbyl sulfonic acids include alkylbenzenesulfonic acids such as p-toluenesulfonic acid and lower Alkanesulfonic acids, e.g. methanesulfonic acid.

Suitable methods of deacylating cephalosporin derivatives containing 7β-acylamido groups are described in British Patent Nos. 1,041,985 and 1,119,806, in Belgian Patent Nos. 719 712 and in South African Patent Nos. 68/5048 and 68 / 5327. Another N-deacylation method that can be used is acid catalysis. For example, the N-deformylation of a 7.beta.-formamido group with a mineral acid at a temperature of- 15 °, are preferably carried out at a temperature of +15 to +40 ⁰ C to + 100 ⁰ C. The N-deformylation can with the help of a Lewis acid in a lower alkanol, preferably un-. 'under substantially anhydrous conditions.

109843/1954

21U541

D. Other reactions

The method according to the invention creates a connection between the formula

R ²

• 4
Q-CH ₂ -C-CO-R XIII

R ³

If the group R contains an esterified carboxyl group, this group can, if desired, for example by hydrolysis a carboxy group can be saponified. The hydrolysis can be carried out under mild conditions by treatment with a base or a Acid. The course of hydrolysis can be determined by paper chromatography to be tracked.

Compounds of the general formula

R ²
Q-CH ₂ -CH-CO-R ⁴ XIV

2 4
wherein Q, R and R have the meanings given above, can be prepared by adding a compound of the general formula

R ²

ι. Q-CH ₂ -C-CO-R XV

COOR ¹⁰

2 4 wherein Q, R and R have the meanings given above

10

and R is hydrogen or an alcohol or phenol radical, decarboxylated.

The decarboxylation is normally carried out at a temperature of, for example -40 ° to +100 ⁰ C, and may together with the cleavage, such as hydrolysis, in the same reaction medium, preferably at a temperature of 0 to 80 ° C be effected · The progress of the decarboxylation can by thin layer chromatography, paper chromatography, NMR spectroscopy and

109843/1954

Determination of the developed CO _{2 can be} tracked.

[Compounds of the formula XV in which R = H can be selected from the corresponding esters or nitriles, 25.B. Connections of general Formula

R ² Q-CH ₅ -C-CO-R ⁴ XVI

2 4
wherein Q, R and R have the meanings given above

and P is CN or COOR, in which R ^{10 is} an ester radical.

Compounds of the formula XVI can in turn be prepared by the carbanion reaction C described above. In addition can be a compound of the formula

Q-CH ₂ X IX

wherein Q and X have the meanings given above, with a carbanion of the formula

R ²

© C-CO-R ⁴ P

2 4
wherein R, R and P have the meanings given above,

are reacted, the reaction is preferably carried out under such conditions that the group of the compound of the formula IX _{exists in the 4-position as CO 2} "when X = Cl.]

If the resulting compound has a sulfinyl group in the 1-position, this can be done by any suitable method be reduced. This can be done, for example, by reducing the corresponding Acyloxysulfonium or. Alkyloxysulfonium salt found in

109843/1954

21U541

situ, e.g. in the case of the acetoxysulphonium salt with acetylene chloride, the reduction e.g. or iodide ion, for example in a solution of potassium iodide in a water-miscible solvent, e.g. Acetic acid, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction can take place at one temperature from -20 ° to +50 ° C.

Alternatively, the reduction of the 1-sulfinyl group with phosphorus trichloride or tribromide in solvents such as methylene chloride, dimethylformamide or tetrahydrofuran, preferably at a temperature of -20 ° C to +50 ⁰ C, can be effected.

If the resulting connection is a Ceph-2-em connection, the desired Ceph-3-am compound can be obtained by treating the the former with a base, e.g. a base of the type used to form carbanions.

Groups which protect amino or carboxyl groups can be removed as described above.

administration

The compounds of the invention can be administered in any suitable manner analogous to other antibiotics are formulated, and the invention also includes pharmaceutical compositions comprising a compound of the general Formula I or a non-toxic derivative, e.g. a salt, thereof (as defined above) and those for use adapted in human or veterinary medicine. Such compositions can with the help of the necessary pharmaceutical Carriers or binders can be prepared in the usual way.

The present invention therefore provides pharmaceutical compositions, which a compound of formula I or a non-toxic derivative thereof (as described above) together with a pharmaceutical carrier or binder. The-

109843/1954

These compositions can be in a form suitable for absorption in the gastrointestinal tract. Tablets and Capsules for oral administration can be in unit dosage form and conventional auxiliaries such as binders, e.g. Syrup, acacia, gelatine, sorbitol, tragacanth gum or polyvinylpyrrolidone} Fillers, e.g. lactose, sugar, corn starch, calcium phosphate, Sorbitol or glycine; Lubricants, e.g., magnesium stearate, talc, polyethylene glycol, silicon dioxide; Dividing agent, e.g. potato starch or compatible wetting agents such as Sodium lauryl sulfate. The tablets can be good known procedures are covered. Orally to be administered liquid preparations can be in the form of aqueous or oily suspensions, solutions, emulsions, syrups, elixirs etc. or can be present as a dry product that mixed with water or other suitable vehicle prior to use. Such liquid preparations can Usual additives such as suspending agents such as sorbitol syrup Methyl cellulose, glucose / sugar syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats / emulsifiers such as lecithin, sorbitan monooleate or acacia; non-aqueous vehicles, which may include edible oils, e.g., almond oil, fractionated coconut oil, oily esters, propylene glycol, or ethyl alcohol; Preservatives, e.g. Methyl or propyl p-hydroxybenzoate, or sorbic acid. Suppositories contain common suppository bases, e.g. cocoa butter or others Glycerides. . .

Compositions for injection may be in unit dosage forms in ampoules or multiple dose vials with a preservative are offset. The compositions can be in the form of suspensions, solutions, emulsions in oily or *. aqueous carriers and can be formulating agents such as suspending agents, stabilizers and / or dispersants, * Alternatively, the active ingredient can be in powder form so that it can be mixed with a suitable carrier material before use. e.g. sterile, pyrogen-free water mixed with

^Be "ORIOJNAt INSPECTED

109843/1954

The compositions can also be prepared in a suitable form to penetrate through the mucous membranes of the nose and the Pharynx or bronchial tissue too, and can conveniently take the form of a powder or liquid Spray or inhalant, take the form of lozenges, throat brush solutions, etc. For treatment of the eyes or ears, the preparations can be in the form of individual capsules, in liquid or semi-solid form, or can be used as drops etc. For topical application, the compositions can be in hydrophobic or hydrophilic bases can be formulated as ointments, creams, lotions, liniments, powders etc.

For veterinary medicine, the composition can e.g. intramammary preparation can be formulated with either slow-acting or fast-acting bases.

The compositions may contain the active material from 0.1 % upwards, preferably in an amount from 10 to 60%, depending on the route of administration. When the compositions are in unit dosage form, each dose preferably contains 50 to 500 mg of the active ingredient. The dosage for the adult human is preferably in the range of 100 to 3000 mg, for example 1500 mg per day, depending on the route of administration and the frequency of administration.

The compounds according to the invention can be administered in combination with other therapeutic agents, such as antibiotics, for example other cephalosporins, with penicillins or tetracyclines.

The following examples are intended to explain the present invention further without, however, restricting it.

In the examples, unless otherwise stated:

109843/195 A

21U541

1) Ultraviolet spectra (UV) measured on solutions in ethanol.

2) Infrared spectra (IR) were measured in Nujol on KBr plates (mulls).

3) The optical rotation values were determined at 19 to 30 at concentrations in the range from 0.5 to 1.5 % as solutions in dimethyl sulfoxide. When other solvents were used, the same concentration range was used.

4) The solutions were dried over anhydrous magnesium sulfate.

5) All types of silica gel were from Merck A. G., Darir.stadt, Germany, related.

6) The nuclear magnetic resonance (NMR) spectra were determined at 60 or 100 MHz. The signs of the coupling constants (J) are not assigned. The signals are g sharp singlets Cs), doublets (d), double doublets (dd), triplets (t), Quartets (q), double quartets (dq), AB-quartets (AB-q), quintets (qu) and multiplets (m).

System A = descending paper chromatography using n-propanol / water = 7/3 on Whatman Nr.l paper at room temperature.

System B uses n-butanol / ethanol / water = 4 / l / 5, equilibrated at room temperature, with the upper phase in equilibrium with the lower phase in descending order, and Whatman 1 or 3MM paper is used which is mixed with 0.05m sodium dihydrogen phosphate is buffered to a p "value of 6.

System C is used Xthylacetat / n-butanol / o, Im-sodium acetate = 8/1/8, p _H value "5, equilibrated at 38 ° C, the upper phase at 38 ° C in equilibrium with the unte ren phase, is used wherein No.1 Whatman paper, Im-sodium acetate to a _pH value of 5 buffered with 0 serving as the developer in a descending manner.

1Q9843 / 19E4

21U541

The petroleum ether used has a boiling point of 40 to 60 ⁰ C. methylene chloride was dried over basic alumina Woelm I. Thin layer chromatography was performed upwards on Merck silica plates, developing with benzene / ethyl acetate = 4/1 or under the following conditions:

System D Merck GF ~ C _{4 + 3} /: _β plates, the upper phase of solvent mixture B being used for developing.

System E is carried out on the plates of system D with benzene / ethyl acetate = 5/1 as developing agent.

System F like system E, but with benzene / ethyl acetate = l / l as solvent.

The following abbreviations are used for the appearance of the stains used: s = strong, m = medium, f = weak, ν = very weak.

The NMR spectra were determined at 60 and 100 MHz. The characters the coupling constants could not be assigned.

As far as possible, the analytical values for the solvates were carried out Examination of the corresponding characteristics in the spectra confirmed.

R _fc means the R _f value divided by that of 3-acetoxymethyl-7β- {2-thienylacetamido) -ceph-3-em-4-carboxylic acid.

The electrophoresis conditions are those described by Cocker et al., In J. Chem. Soc., 1965, 5015.

The organic solutions were dried over dried magnesium sulfate.

109843/1954

example 1

a) Diphenylmethyl-3- (2 ^ ~ methyl-3-oxopentyl) -7ß- (2-thienylacet-

¹ ir - ι ι 'Jf

amido) ~ ceph-3-em-4-carboxylate

A solution of diphenylmethyl-3-iodomethyl-7ß- (2-thienylacetamido) -ceph-3-em ~ 4 ~ carboxylate (8g, 12.8 mmol) in 150 ml of dry benzene was mixed with 8 ml of freshly distilled N- (Pent- 2-enyl) pyrrolidine (approx. 4 equivalents) and after 2 minutes at room temperature with a mixture of glacial acetic acid (25 ml) in 140 ml of water, followed by 75 ml of acetone. The mixture was shaken until all of the oil had dissolved, then the organic layer was separated and the aqueous layer was extracted with ethyl acetate. The organic extracts were combined, dried by passing over magnesium sulfate and the volume was reduced to 25 ml. This solution was chromatographed on 0.05-0.2 mm silica gel in benzene / ethyl acetate (10/1), and the fractions which contained a substance which gave a spot in thin-layer chromatography (silica gel GF ₃₅₄ in benzene / ethyl acetate, 5 / l), R _f value approx. 0.4, were collected and evaporated to a foam (4.8 g, 60%). The foam was dissolved in ethanol, from which the diphenylmethyl ester η colorless prisms abschied (1.5 g, 20%), m.p. = 147-150 ⁰ C, [a] _D + 26 ° (tetrahydrofuran), UV spectrum λ _{χ f} , 258 nm (6 6200), IR spectrum

\> (CHBr-.) 3680 (H ₀ O). 3380 (NH), 1770 (β-lactam), 1715

max. j ^ λ

(COOR), 1700 (C = O) and 1670 and 1505 cm (CONH), NMR spectrum (CDCl ₃ ) Z 2.68 [(Ph) ₂ ) CH-], 3.08 to 2.73 (3- Proton m, 2-thienyl), 3.09 (Ph ₃ CH-), 3.32 (NH), 4.28 (dd, 7-H), 5.10 (d, J = 4.5 Hz, 6 -H), 6.18 (2-thienylmethyl), 6.68 and 6.93 (AB-q J = 18 Hz., 2-H) 7.2 to 7.8 (5 proton envelope, CH ₂ CH (Ct 9.04 (CH ₂ CH ₃ ) and 9.14 OCH-CH ₃ , C ₃₃ H ₃₂ N ₂ O ₅ S ₂ '-! / ^ H ₃ calculated: C 64.2 H 5.5 N 4, 7 S 10.7% found: 64.2 5.3 4.5 10.7%.

b) Sodium 3- (2 ^ -methyl-3-oxopentyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylate

A solution of diphenylmethyl 3- (2-methyl-3-oxopentyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (2 g, 3.4 mmol)

109843/1954

in 8 ml of trifluoroacetic acid and 3 ml of anisole was allowed to stand for 5 minutes at room temperature, and then the reagents were separated under reduced pressure. The resin was dissolved in ethyl acetate and poured into Petrolättier (Kp. 60 to 80 ⁰ C). The pale yellow solid was dissolved in 5 ml of ethyl acetate and 20 ml of ether and treated with 10% sodium 2-ethylhexanoate in 5 ml of acetone. The sodium salt separated as. Pale yellow amorphous solid from (700 mg, 46%), [ücJ _d + 65 ° (H ₀ O), UV spectrum λ (H ₀ O) 234, 260 nm (£ 13,200 or 7900

IR spectrum V 3450 (H _o 0), 3290 (NH), 1764 (ß-lactam),

^ max. έ

1700 (C = O), 1680 and 1540 (CONH), and 1600 cm (COO * "), NMR spectrum (D ₂ O), T 2.65 to 2.98 (3-proton m, 2-thienyl ), 4.48 (d, J »4.5 Hz., 7-H), 4.98 (d, 6-H) 6.13 (2-thienylmethyl), 6.49 and 6.90 (AB- q, J = 18 Hz, 2-H), 6.91 to 7.82 (5-proton envelope; CH ₂ CH (CH ₃ ) COCH ₂ CH ₃ ), 8.92 O CHCH ₃ ) and 9.05 ( CH ₂ CH ₃ ) ^C 19 ^H 21 ^N 2 ^NaO 5 ^S 2 * ^2H 2 ° ^{calculated: c 47} > ^{5 H 5} » ^{2 N 5} » ^{8 S 13} > ⁴ *

found: 47.2 4.9 5.5 13.3 %

(System C).

Example 2

a) Diphenylmethyl-7β-amino-3- (2 ^ -methyl-3-oxopentyl) -ceph-3-em-4-carboxylate-hydrogen-p-toluenesulfonate

A solution of diphenylmethyl 3- (2 ϊ -methyl-3-oxopentyl) -7ß- (2-thienylacetmido) -ceph-3-em-4-carboxylate (1.6 g, 2.72 mmol) and 2.7 ml of pyridine (33.5 mmol) in methylene chloride (30 ml) was cooled to -10 ° C. and treated with a solution of phosphorus pentachloride (1.7 g, 8.15 mmol) in 35 ml of methylene chloride over 15 minutes. The solution was stirred at -12 ° C. for 45 minutes. Then, methanol was added (27 ml) at such a rate that the temperature did not rise above -10 ° C, and after completion of the addition, the temperature was raised to 20 ⁰ C. After standing for 3 hours, the solution was ^{cooled to 0} ° C. and treated with 30 ml of water with vigorous stirring. The temperature was then increased to 20 ° C. and stirring was continued for a further 30 minutes. The organic layer was separated with

109843/1954

aqueous sodium bicarbonate and then dried and evaporated. A solution of the resin in ethyl acetate (20 ml) and ether (30 ml) was treated with p-toluenesulfonic acid hydrate (0.6 g, 3.2 mmol) in 10 ml of ethyl acetate. The toluenesulfonate separated out in the form of colorless needles (1.21 g, 69.5%),? = 174 to 176 ° C (dec.), [A] _n + 5 ° (CHCl,), UV spectrum 259 nin (£ 6800), JR spectrum ν 1778 (β-lactam), 1718 (COOH) and 1703 cm "(C = O), NMR spectrum (dg-dimethyl sulfoxide)

t 2.60 (Ph _-2 CH-), 2.47 and x 2.86 (£ CH ₃ -CgH ₄ SOp, 3.07

4.68 (dd, 7-H), 4.86 (d, J - 4.5 Hz, 6-H), 6.42 (broad s, 2-H), 7.2 to 7.6 (5th - Proton Envelope _s CH _-2 CHjCH ₃ ) COCH ₂ CH ₃ ), 7.70 (2-CH ₃ C ₆ H ₄ -), 9.10 CCH ₂ CH ₃ ), 9.14 (> CHCH ₃ ), C ₃₃ H ₃₆ N ₂ S ₂ -1 / 2H ₂ O

Calculated: C 61.4 H 5.8 N 4.3 S 9.9% found; 61.8 5.7 4.2 10.2 %

b) 7β- (Da-aminophenvlacetmido) -3- (2 | - methyl-3-oxopentyl) -ceph-3-ero-4-carboxylic acid trifluoroacetate

A solution of N- (tert-butoxycarbonyl) -D-phenylglycine (0.93 g, 3.86 raffol) in tetrahydrofuran was cooled to -6 C and ^0, followed by 0.55 ml triethylamine (3.86 mmol) of 0.53 g of isobutyl chloroformate (3.86 mmol) treated, the temperature being kept below -6 ⁰ C. After 30 minutes the triethylamine hydrochloride was filtered off and the filtrate was added to a solution of diphenyl ethyl-7β-amino-3- (2 ^ -methyl-3-oxopentyl) -ceph-3-em-4-carboxylate-hydrogen-p- toluenesulfonate (1.6 g, 2.5 mmol) in 10 ml of acetonitrile and 5 ml of Ν, Ν-dimethylacetamide. After 30 minutes the acetonitrile was removed under reduced pressure, "The residue was dissolved in ethyl acetate, una washed with aqueous sodium bicarbonate and water, then dried and evaporated. The resin was treated with 4 ml of anisole and 10 ml of trifluoroacetic acid, and after 5 minutes, the reagents were removed under reduced pressure · The resulting oil was suspended in 50 ml of water and ion-exchanged Amber lit LA 1 (OAc ") in ether (10% _e _. 50 ral) foehand 'delt. After shaking, the aqueous layer was separated off and washed with ether and ethyl acetate and freeze-dried

10 9 8 4 3/1954

and gave the title compound in the form of a pale yellow solid (0.92 g, 85%), [a] _D + 75 ° (acetone / water «l / l), UV spectrum X _max (H ₂ O) 261 nm (6 7100), IR spectrum ^ _maXm 1756 (ß-lactam), 1680 (CONH), 1620 cm * ¹ (C00 ~), NMR spectrum (D ₂ O), τ 2.50 (C ₆ H ₅ ), 4 .39 (d, J = 4.5 Hz, 7-H), 4.92 (d, J «4.5 Hz, 6-H), 4.75 [C ₆ H ₅ CH (NH ₃ )], 6.55, 6.92 (AB-q, J = 18 Hz, 2-H), 6.90 to 7.55 (5 proton envelope [CIi ₂ CH (CH ₃ ) COCH _-2 CH ₃ ], 8 , 96 (> CHCH ₃ ) and 9.04 (CH ₂ CH ₃ ). R _f 0.15 (system B), 0.08 (system C). -. ·

c) 7β- (D-2-aminophenylacetamido) -3- (2i-methyl-3-oxopentyl) -ceph-3-em-4-carboxylic acid

A solution of N-itert-butoxycarbonyD-D-phenylglycine (5.83 g, 22 mmol) in 60 ml of dry tetrahydrofuran was cooled to -10 ° C and triethylamine (3.05 ml, 22 mmol) followed by isobutyl chloroformate (3.004 g, 22 mmol) treated in 10 ml of dry tetrahydrofuran, the temperature being kept below -6 ⁰ C. The temperature was then increased to 20 ° C. and the white suspension was stirred for 30 minutes. The precipitated triethylamine hydrochloride was filtered off and the filtrate was added to a stirred solution of diphenylmethyl-7β-amino-3- (2 | -methyl-3-oxopentyl) -ceph-3-em-4 in the course of 5 minutes Carb ~ oxylate hydrogen p-toluenesulfonate (9.11 g, 14.3 mmol) in 60 ml of acetonitrile and 30 ml of Ν, Ν-dimethylacetamide. After 30 minutes, the solvents were removed at 40 ° C. (2 mm Hg) and the residue was dissolved in 250 ml of ethyl acetate. The solution was washed with aqueous sodium bicarbonate (2 x 50 ml) and water (2 × 50 ml) and brine (50 ml), dried and evaporated to give a pale yellow foam (10.68 g). The foam was treated with 20 ml of anisole and 50 ml of trifluoroacetic acid, and after 5 minutes the reactants were ^{distilled off at 30 °} C. (2 mm Hg). The resulting oil was suspended in 800 ml of water and treated with an ion exchanger Amberlit LA 1 (OAc ") in Xther (10%, 500 ml). After shaking, the insoluble material (440 mg) was filtered off and the aqueous layer separated and washed with ether (3 200 ml) and ethyl acetate (3 χ 200 ml)

109843/1954

and then freeze-dried to give a pale yellow solid (5.74 g). This material was triturated with 200 ml of ether and gave the title compound (5.315 g, 86%) in the form of a pale yellow solid, [oc] _D + 71 ° (acetone / water - l / l), UV spectrum λ _v (0, 1 m «-p» 6-phosphate buffer),

262.5 nm ( t 6600), IR spectrum V 1760 (ß-lactam),

- IUcLX *

1700 (ketone) and 1600 cm (-CO ₂ ""), NMR spectrum (D 0 with trifluoroacetic acid) 2.50 (CgH ₅ -), 4.39 (d, J 4.5 Hz, C ₇ -H) , 4.92 (d, J 45 Hz, C ₅ -H), 4.75 (Ph CH-), 6.62 and 6.85 (AB-q, J 18 Hz, C ₂ -H), 6, 90 to 7.60 (CH ₂ CH (CH ₃ ) COCH_ ₂ CH ₃ ), 8.84 to 9.10 (envelope, -COCH ₂ CH _-3 and> CHCH_ ₃ ). ^c ₂ i ^H 25 ^N 3 ° 5 ^S calculated: C 58.5 H 5.8 N 9.7 S. 7.4% found: 56.8. 5.9 6.3 6.3%

Example 3

3- (2r-Ethoxycarbonyl-3-oxobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid

A solution of ethyl acetoacetate (17.5 g, 135 mmol, 5 equivalents) in freshly dried peroxide-free tetrahydrofuran (50 ml) was added dropwise with stirring at a temperature below 35 ° C. to a suspension of sodium hydride (3.81 g of a 45% - igen dispersion in mineral oil, 71.5 mmol, 2.7 equivalents) in 10 ml of tetrahydrofuran. After 30 minutes the addition was complete and the solution was stirred for a further 15 minutes and then added dropwise with stirring over 25 minutes at a temperature below 17 ° C. to a solution of 3-chloromethyl-7β- (2-thienylacetamido) -ceph -3-em-4-carboxylic acid (9.8 g, 26.3 mmot! In 150 ml of freshly dried tetrahydrofuran. The pale orange-red solution was stirred for a further 30 minutes at 17 ° C. and then added to 100 ml of in-hydrochloric acid . the tetrahydrofuran was evaporated, brought the _pH value to 8.0 and the solution was washed with 'ethyl acetate. the aqueous layer was acidified to a _pH value of 2.0 and extracted with ethyl acetate, which was washed, dried, partially was evaporated and added to a large volume of petroleum ether (bp. 60 to 8O ⁰ C),

1098 43/19 54

ORIGINAL INSPECTED

.41-21U541

almost white flakes resulted (8.6 g, 70%), F = up to 80 ⁰ C (decomp.), [<x] _D + 36 ° (CHCl ₃ ), UV spectrum λ 238 nm (£ 12000) , Inflexion at 260 nm (£ 6600); a sample purified over sodium salt had an F = 115 to 125 ° C (decomp.), UV spectrum λ _χ (0, Im phosphate, p _H 6) 236 nm (ε 13800), inflection at 260 nm, (£ 8350 ), IR spectrum V _{m = v} (CHBr.,), 3400 (NH), 1775 (β-lactam), 1730 to 1700 (CO ₂ R, CO ₂ H and COMe), 1680 and 1510 cm " ¹ (CONH ), ~ c (60 MHz; and 100 MHz; D ₂ O-NaHCO ₃ ) 2.65 to 3.03 (about 3-proton m; 2-thienyl)., 4.47 (1-proton d, J 5 Hz; C ₇ -H), 5.0 (1-proton d, J 5 Hz; Cg-H), 5.85 (2-proton m; OCH ₂ -Me), 6.12 (2-proton s; -CH ₂ -CO) 6.49, 6.45 and 6.85 (2 protons, 2 superimposed AB-q, J 17 Hz; C ₃ -CH ₂ ) 6.72, 6.78 and 7.39 ( two superimposed AB-q, J 14 Hz; C ₃ -CH ₃ ), 7.71 and 7.74 (3 protons, two s; COMe), 8.76 (3 protons, 2t, J 7 Hz, um about 0.5 Hz separated); 0-CH ₃ -CH ₃ ) JC ₃₀ H ₃₂ N ₃ O ₇ S ₂ (466.53) calculated: C 51.5 H 4.9 N 6.0 S 13.75%

found: 51.7 5.0 5.6 13.2 and 13.1%, R 0.6 (system C, 0.2 mg, loading), 0.6 (system B, 0.2 mg, Load), 0.45 (system D, 0.1 mg, load). The compound gave with a 1% methanolic iron (III) chloride solution a reddish brown color.

Example 4

a) Diphenylmethyl 3- (3-oxobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate

3- (2 ^ -ethoxycarbonyl) -3-oxobutyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid (6.3 g) was dissolved in 100 ml of water containing 6.5 g Containing potassium carbonate, and kept for 5 hours at 50 ° C, after which the paper chromatography (system C, product R_ 0.25, starting material R _f 0.6) indicated an almost complete consumption of the starting material. The red solution was washed with ethyl acetate, acidified to a pn value of 1.5 and extracted with ethyl acetate which was washed with water and dried and evaporated to give a foam (4.0 g). A solution of the foam in 40 ml of freshly dried tetrahydrofuran was added with 3 g

109843/1964

21U541

Diphenyldiazomethane is esterified in 30 ml of petroleum ether (b.p. 60 to 80 ° C) for 16 hours at 22 to 25 C. The solvents were then evaporated and the red oil was dissolved in ethyl acetate and added to an excess of petroleum ether to give a pale brown powder (4.98 g). This powder was chromatographed on a silica gel column (0.05-0.2 mm) to give the title compound (1.8 g, 24% yield from the product of Example 3).

A sample was recrystallized from ethyl acetate and gave prisms, F = 15 7 to 159 ° C (dec.), [A] _D + 4.3 ° tetrahydrofuran), UV spectrum A. _max 235 nm ( L · 13300) and inflection at 260 nm (67,100), IR spectrum V * (CHBr.,) 3420 (NH), 1780

ΓΠα.Χ · J ^

(lj-lactam), 1715 (CO ₂ R and COMe) and 16 80 and 1510 cm (CONH) V max. 3310 (NH), 1755 (ß-lactam, 1715 and 1705 (CO ₂ R and COMe), 1670 and 1525 cm " ¹ (amide), t (CDCl ₃ ) 2.6 to 3.1 (m, 2-thienyl), 3.14 (1-proton s, CHPh ₃ ), 3.6 (! -Proton d, J 9 Hz, CONH), 4.25 (C ₍₇₎ -H, dd, J 5 and 9 Hz), 5.1 (C ₍₆₎ -H, d, J 5 Hz) 6.18 (2- proton s, CH ₂ CO), 6.56, 6.82 (C ^ ₂ J-CH _2, AB q, J 18 Hz), 7.2 to 7.7 (4-proton m, possibly A ₃ B ₂ pattern; 3-CH ₂ 'CH ₃ = CH), 7.98 (3-proton s, COMe), the deuteration only changed the signals for the amide NH and the proton in the 7-position, the former was washed out and the latter was converted to a doublet, Cj-HpoNpOj-Sp: calculated: C 64.3 H 5.05 N 5.0 S 11.4% found: 64.1 5.1 4.6 11.8 % R _f 0.35 (system E).

b) Diphenylmethyl-7β-amino-3- (3-oxobutyl) -ceph-3-em-4-carboxylate-p-toluenesulfonic acid salt

A solution of the ester (1.33 g from Example 4a) in 25 ml of dry methylene chloride (2.35 g, 29.8 mmol) with 2.4 ml of pyridine and cooled to ⁰ C -2.0. Then 1.46 g of phosphorus pentachloride (7.0 mmol, 2.9 equivalents) were added over a period of 13 minutes in 6 additions, the temperature being kept below -20 ° C. The faint orange

109843/1854

21U5A1

The colored suspension was quickly warmed to 0 C, stirred for 15 minutes, cooled to -35 ° C and treated with 25 ml of cold methanol at below -30 ° C. The solution was warmed over 15 minutes to 0 ⁰ C (ice bath) and then at 20 ° C and stirred for 3 hours and 20 minutes. Then, the solution was cooled to 0 ⁰ C and stirred for 1 hour under heating at 25 ° C with 2 N hydrochloric acid. The organic layer was washed with hydrochloric acid, water, sodium bicarbonate, water and brine, dried and evaporated. The resin was dissolved in 5 ml of ethyl acetate and added to a solution of p-toluenesulfonic acid (420 mg) in 5 ml of ethyl acetate. The solution was cooled and 10 ml of ether were added while stirring. After stirring for 2 hours, a white crystalline solid (513 mg, 35%), F − 131 to 139 ° C. (decomp.), UV spectrum \ _{m = iv} 260 mn (£ .6450), IR spectrum V was obtained "(CHBr-,) 1788 (ß-lactan), 1715 (CO ₂ R (, 1230 cm (CO ₂ R), τ (CDCl ₃ 100 MHz), 2.26 and 3.0 (4 protons, 2 d, J 7 Hz; Me CgH ₄ -), 2.75 (10-proton s, Ph ₃ ), 3.19 (1-proton s, CH-Ph ₃ ), 5.17 (2-proton s, C _{(? )} H and C ₍₆₎ H), 6.61 and 7.35 (C ₍₂₎ -CH ₂ , AB-q, J 17 Hz), 7.2 to 7.8 (m, -CH ₂ -CH ₂ -CO), 7.79 (s, about 3 protons; COMe), 8.09 (3-proton s, C ₅ H ₄ -Me), R _f 0.3 (system F).

c) 7β- (D-2-aminophenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid, trifluoroacetic acid salt

A solution of N-tert-butoxycarbonyl-D-phenylglycine (2.735 g, 10.8 mmol, 3.5 equivalents) in dry tetrahydrofuran (20 ml) was mixed with 1.54 ml of triethylamine (1.11 g, 11 mmol) in 10 ml of dried tetrahydrofuran. Then, 1.45 g of isobutyl chloroformate (10.5 mmol) were added and the suspension stirred for 30 minutes at 20 ⁰ C. The suspension was then filtered and added to a solution of the amino ester (prepared as in Example 4 b, 1.82 g, 3 mmol) in 12 ml of acetonitrile and 4 ml of Ν, Ν-dimethylacetamide, and the mixture was for 30 minutes at 20 ⁰ C stirred. Then excess sodium bicarbonate was added and the emulsion was stirred for 1 hour. Ethyl acetate was then added and the mixture was washed with Na-

109843/1954

.- «4- 21H5A1

trium bicarbonate solution, water and saline solution. The solution was dried, evaporated and the residue (2.65 g) over silica gel (0.02 to 0.5 mm) was chromatographed, so that 1.6 g of a chromatographically homogeneous foam were obtained.

The foam was dissolved in 1.5 ml of anisole and 6 ml of trifluoroacetic acid and evaporated after 4 minutes at 20 ° C. at 20 ° C./2 mm Hg. Then 2 ml of ethyl acetate were added and the mixture was triturated with ether to give the title compound (905 mg, 60% yield based on the 7-amino ester) as a pale yellow solid, mp 115.5-136 ° C (dec.), [AL · + 69 °, UV spectrum λ _{= v} (0.1 m-phosphate, p "6) 261 nm (£ 7800),

IR spectrum V _{m = Y} 1780 (ß-lactam), 1720 to 1680 (CO ₀ H,

F ₃ C-CO ₂ ", CONH), 720 cm (Ph), NMR spectrum T (D ₂ O, containing

F ₃ CCO ₂ H) 2.5 (5-proton s, Ph), 4.38 (C _(?) -H, d, J 4 Hz), 4.79

(1-proton s, Ph ₂ CH (JiH ₃ ) CO), 4.97 (C ₍₆₎ -H, d, J 4 Hz), 6.52 and

6.85 < ^C (2) - ^CH 2> ^AB -q » ^{J 18 Hz)} » ⁷ » ^{2 to 7} » ⁵ (4-proton m, 3-CH ₂ -CH ₂ -CO-), 7.84 ( 3-proton s, COMe); Ethyl acetate (about 0.1 mole) was also present. R, 0.25 (system C, R, 0.5 (system B), R _f 0.1 (system D). The product migrated with a distinct positive charge at p "1.9.

Example 5 3- (3-Oxobutyl) -7ß- (2-thienvlacetamido) -ceph-3-em-4-carboxylic acid

A solution of diphenyl methylacetoacetate (810 mg, 3.1 mmol) in 6 ml of freshly dried peroxide-free tetrahydrofuran was added to 142 mg of sodium hydride (45% suspension, 2.65 mmol), a clear pale yellow oil solution being obtained with stirring in 5 minutes e ner _A solution of 3-chloromethyl-7ß- (2-thienylacetamido) ceph-3-em-4-carboxylic acid (383 mg, 1.01 mmol) was added in 4 ml of tetrahydrofuran. After about half of the reagent was added to the reaction mixture, a gel formed. A further 2 ml of tetrahydrofuran were then added and the addition was ended with vigorous stirring. The color changed from yellow to orange. After 30 minutes at 25 ° C, the red solution became excess

109843/1954

OOPY

21H541

Given hydrochloric acid. [Thin layer chromatography (system D, with a new spot at R _f 0.55) showed almost complete reaction after 5 minutes]. The tetrahydrofuran was then evaporated and partitioned the reaction product between water and ethyl acetate at a p ~ value of _H. 8 The organic layer was washed, dried and concentrated and added dropwise to petroleum ether (b.p. 60 to 80 ° C.), a pale yellow powder A (254 mg) being obtained; the petroleum ether contained essentially unchanged diphenyl methylacetoacetate (shown by NMR and IR spectra). The aqueous phase was acidified and extracted with ethyl acetate, which was dried and evaporated to give a red oil

- (100 mg) which did not contain a strong maximum at around 1780 cm (CHBr-) and in the NMR spectrum did not show any protons with E values between 3.8 and 5.4 (which are usual for those of the ß-lactam- Rings are). Powder A (188 mg) was dissolved in 0.2 ml of anisole and 0.8 ml of trifluoroacetic acid and evaporated after 4 minutes in a rotary evaporator at 40 c / 2 mm Hg. The oil was dispersed at a _pH value of 8 between water and ethyl acetate. The aqueous layer was acidified to a p "value of 2.0 and extracted with ethyl acetate, which was washed, dried and converted to a brown foam, R _f 0.9 (m, streaky), 0.29 (s), 0, 1 (f) (System C) which was purified by preparative paper chromatography (System C) to give the title compound (20 mg), NMR spectrum ZT (D ₂ O, with NaHCO ₃ ) 2.6 to 3.1 (3-proton m, 2-thienyl), 4.5 (1-proton d, J = 5 Hz; 7-H), 5.0 (1-proton d, J = 5 Hz; 6-H), 6 , 15 (2-proton s, CH ₂ CO), 6.5 and 6.90 (2 protons, 2 d, J = 18 Hz, 2-CH ₃ ), about 7.2 to 7.6 (4-proton m, possibly ApBp pattern; 3-CH ₂ CH ₂ CO), 7.81 (3-proton s, COMe), R _f 0.29 (system C).

Example 6

a) 3-Chloromethyl-7ß- (2-thienylacetmido) -ceph-3-em-4-carboxylic acid

2.223 g of diphenylmethyl-3-chloromethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (4.1 mmol) were suspended in 2 ml of anisole and treated with 8 ml of trifluoroacetic acid for 4 minutes

109843/195 A '

001Ύ

21U541

Treated at 20 ° C. The solution was then evaporated at 40 ° C./2 mm Hg to give an oil which was triturated with ethyl acetate and filtered to give a pale yellow solution. The solution was concentrated to 12 ml and added dropwise with stirring to petroleum ether (500 ml), whereby a pale yellow solid was obtained (1.126 g), \ _mav 235 nm (e} ^% 324),

max. learn *

260 nm (Ejf * 169). From the UV and NMR spectra it was found learn

that the material had a purity of about 80%.

A solution of the yellow solid in ethyl acetate was treated with 5 volumes of ether. The suspension was filtered, decolorized with activated charcoal and concentrated to 2 to 3 ml.

Then 1 ml of ether was added and crystallization was initiated by scratching the inside of the vessel with a spatula triggered by white needles (257mg). The filtrate was added dropwise to petroleum ether to give a solid (373 mg).

The crystalline sample had an F = 40.5 to 51.5 ° C (decomp.), [A] _D -4.45 ° (tetrahydrofuran), UV spectrum X _max (tetrahydrofuran) 269 nm ( t 8600), λ 235 nm (£ 9800) with an in-

⁷ max.

flexion at 260 nm (£ 3700), IR spectrum ^ _max 3310 (NH), 1775 (β-lactam), 1725 and 1698 (CO ₂ H), 1760 to 1655 and 1533 cm " ¹ (CONH), NMR spectrum TT (CDCl ₃ with 2 drops of DMSO-d ⁶ ; 60 MHz) 1.3 (about 1-proton s, CO ₂ H) _r 1.8 (2-proton d, J = 9 Hz; CONH), 2.7 to 3.1 (3-proton in, thienyl), 4.22 (1-proton dd, J = 5 and 9 Hz; 7-H) 4.99 (1-proton d, ^% J = 5 Hz; 6- H), 5.45 (2-proton s, CH ₃ - Cl), 6.20 (2-proton s, CH ₂ CO), 6.45 (2-proton s, 2-CH ₂ ). A sample contained anisole (0.5 mol, by NMR spectrum), C ₁₄ H ^ ₃ ClN ₂ O ₄ S ₂ .l / 2 PhOMe calculated: C 49.0 H 4.0 N 6.5 Cl 8.3% found: 48 .7 4.4 6.1 8.8%

The material had the following chronatographic properties: R _f 0.68 (m), 0.45 (m), 0.4 (m), 0.2 (m) (system D);

103843/1954

.47-21U541

0.93 <s), 0.07 (m) (system C); 0.55 to 0.3 (m), 0.16 (m) (System B). The compound is unstable in aqueous and alcoholic solvents.

b) 3- (2-Acetyl-3-oxobutyl) -7β-(2-thlenylacetamido) -ceph-3-etn-4-carboxylic acid

7.223 g of acetylacetone (72.2 mmol, 5 equivalents) were dissolved in 65 ml of freshly dried peroxide-free tetrahydrofuran and slowly added to 1.95 g of sodium hydride (45% dispersion, 36.7 mmol, 2.5 equivalents) with stirring at 30.degree ) given in 60 ml of freshly dried tetrahydrofuran. The white suspension was added dropwise with stirring to 5.4 g of 3-chloromethyl-7β- (2-thienylacetamido) -ceph -3-em-4-carboxylic acid (14.5 mmol) in 125 ml of freshly dried tetrahydrofuran over 10 minutes given at 25 ° C. After 90 minutes the reaction mixture was poured onto 40 ml of 1N hydrochloric acid and evaporated to separate the tetranydrofuran. Then Xthylacetat was added and the solution at a _pH value of 8.0 · washed The aqueous layer was acidified to a _pH value of 2.0 and extracted with ethyl acetate, which was washed, dried and evaporated, so that 4 g of a foam was obtained. The foam was then dissolved in acetone and treated with sodium 2-ethylhexanoate (1 equivalent in acetone, 10 ml). Slow addition of ether gave the title compound in the form of the sodium salt . F = 123 to 130 ° C (decomp.), [A] _n -43.8 ° (water), UV spectrum X _mev (0.1m - p "6 phosphate) 236 nm ( t 14000) and 263 nm ( £ 8000) (The addition of 1 drop of In sodium hydroxide gave a new maximum at 312 nm ( t ο 000), which could be ascribed to an enolate chromophore). IR spectrum V (CHBr-,) 1755 (β-lactam). 1690

max . j »■

(COMe, amide), 1520 (amide) cm, NMR spectrum t: (D ₃ O with NaHCO ₃ ) 2.6 to 3.0 (3-proton m, thienyl), 4.47 (1-proton d, J * 5 Hzj 7-H), 4.99 (1-proton d, .J »5 Hz; 6-H), 6.12 (2-proton s, CH ₂ CO), 6.5 and 6.88 (about 2 protons, 2 d, J - 18 Hz; 2-CH ₂ ), 6.81 and 7.43 (about 2 protons, 2 d, J = 14 Hz; 3-CH ₂ ), 7.71 and 7 , 75 (about 5.5 ptotons, 2 s, possibly 2 magnetically non-equivalent COMe groups), 7.88 (about 0.5 proton,

109843/1954

- * 48 -

possibly -C = C-Me of the enol form); the NMR spectrum in DpO was less clear. In CDCl., The spectrum of the acid was complex, showing 30 to 40 % of the enol form. C ₁₉ H ₁₉ N ₂ NaOgS ₂ * 1/2 H ₃ O: calculated: C 49.0 H 4.3 N 6.02 Na 4.9 S 13.7% found: 48.8 4.2 "5, 6 5.0 '13.4% chromatography R _f 0.43 (system B), 0.13 (f), 0.38 (s) (system C), 0.40 (system D) The compound gave with a 1% solution of anhydrous ferric chloride in methanol turns brown.

Example 7

3- (2 fr - tert-butoxycarbonyl-3-oxobuty1) -7β- (2-thienylacetamido) - ceph-3-em-4-carboxylic acid

A solution of tert-butyl acetoacetate (7.914 g, 50 mmol, 3.4 equivalents) in 25 ml of freshly dried tetrahydrofuran was added with stirring at less than 35 ° C. to a suspension of sodium hydride (1.63 g of a 40% strength dispersion , 2.7 mmol, 1.85 equivalents - erroneously insufficient amount of the material, since 2.3 to 2.7 equivalents of the material must normally be used) in 5 ml of tetrahydrofuran. The solution was stirred for 20 minutes at 20 ⁰ C and then added dropwise at 20 ° C to a stirred solution of 3-chloromethyl-7ß- (2-thienylacetamido) ceph-3-em-4-carboxylic acid (5.454 g, 14.6 mmol) in 85 ml of tetrahydrofuran. The addition was made in 15 minutes and the reaction mixture was stirred for an additional 35 minutes before pouring into 1N hydrochloric acid (50 ml, excess).

The tetrahydrofuran was evaporated and the aqueous layer washed with a p "value of 7.8 with ethyl acetate before it was extracted at a p _H ~ value of 2.5 in ethyl acetate. The organic layer was washed, dried and steam t ₃ so that 4.988 g obtained a foam which was dissolved in ethyl acetate and added to a large volume of petroleum ether (bp. 60 to 8O ⁰ C), so that the keto ester in the form of a almost colorless powder obtained (4.084 g, 57%), F = 80 to 92 ° C (dec.) ,.

109843/1954

© BIGINÄL INSPECTED

- 49 [α] _η + 25 ° (CHCl.,), UV spectrum K _{1n αν} (O, lm-p 6 phosphate)

Lj ό ΓΠ3.Λ · ΓΙ

236.5 nm (£ 13600), inflection at 250 nm (ε 8050), ^ _max 23 7 ηπι (£ 14000), inflection at 260 nm (£ 7200), IR spectrum ^ν max ^35Ο ° ^(Η 2 ⁰⁾ » ³³ ° ^(ΝΗ) > ¹⁷⁷⁶ (ß-lactam), 1728 ₂ 1710 (COMe) 16 70 and 1530 cm " ¹ (CONH), NMR spectrum X> (100 MHz; sSmtl. DO - NaHCO,); 2.7 up to 3.1 (3-proton m; 2-thienyl), 4.41 (1-proton, 2 d, J 5 Hz; C ₇ -H), 4.8 (0.5 proton s; not assigned), 4.98 (! -Proton 2 d, J 5 Hz; C ₅ -H), 6.11 (2-proton broad s; CH ₂ CO), 6.2 to 7.5 (about 4 poorly resolved protons m; C ₂ -CH ₂ and C ₃ -CH ₃ ), 7.70 and 7.73 (2.5 protons, 2 s; 2 types of COMe), 7.85 (0.5 proton s; C = C- Me des enol), 8.53 (9-proton s; CO ₂ Bu-t) [C ₂₃ H ₂₆ N ₂ O ₇ SO, 5 H ₃ O (494.58) calcd: C 52.5 H 5.4 N 5 , 6 S 12.8% found: 52.4 5.5 5.3 13.2 %]; R _f 0.45 (system B, 0.2 mg load), R _t 2.2 (system C, 0 , 2 mg load), R 0.25 (f), 0.5 (s) (system D, 0.1 mg load), 0.0 (Merck plate developed with ethyl acetate).

Example 8

3- (2 i - Diphenylmethoxycarbonyl-3-oxobutyl) -7β- (2-thienylacetaroido) -ceph-3-em-4-carboxylic acid

A solution of 12.149 g of diphenyl methylacetoacetate (45.5 mmol, 2.9 equivalents) in 50 ml of freshly dried peroxide-free tetrahydrofuran was added dropwise to a suspension of sodium hydride with stirring at a temperature of 25 to 30 ° C (2.162 g of a 45% dispersion, 40.6 mmol, 2.6 equivalents) in 10 ml of tetrahydrofuran over the course of. Added 20 minutes. The yellow solution became a stirred solution of 3-chloromethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid at a temperature of 25 ° C (5.849 g, 15.7 mmol) in 150 ml of tetrahydrofuran. The addition took place in 25 minutes, and the pale yellow solution was stirred for an additional 25 minutes before pouring into 2N hydrochloric acid (45 ml).

The solution was evaporated and at a p "value of 7.8 with Ethyl acetate extracted. The organic phase was washed,

109843/1954

; -so- 21H5A1 Ty

dried, partially evaporated and added to a large volume of petroleum ether, so that white flakes were obtained (5.947 g, 63%), F = 95 to 106 ° C (decomp.), [α] _β ⁺⁸ ° (CHCl ₃ ), UV spec -. . ν

strand λ inflexion at 235 nm (£ 13500) and 260 nm (£ 8100), Max.

IR spectrum V (CHBr-.) 3600 to 3400 '(H ₀ O), 3300 (NH), -1760 (β-lactam) 1700 to 1670 (CONH, CO ₃ H, COR), 1590 (CONH) and 760 cm " ¹ (Ph), NMR spectrum T (60 MHz; C ₅ D ₅ N) 6.2 (2-proton s; CH ₂ CO), 7.85 (3-proton s) (all other signals were on weak to be able to be assigned), (60 MHz; D ₃ CSOCD,) 2.61 (10-proton s; Ph-), 2.6 to 3.05 (3-proton m; 2-thienyl), 3 , 15 (1 proton s, CHPh ^_2): 4.40 (1 proton not resolved m, C ₇ -H), 5.12 (1 proton not resolved m, C ^ H), 6.2 (2-proton s; CH ₂ CO), 6.2 to 6.8 (C ₂ -CH ₃ , C ₃ -CH ₃ and H _? 0 in D ₃ CSOCD ₃ ), 7.79 (3-proton s; COMe), C ₃₁ H ₃₈ N ₂ O ₇ S ₂ -O ₁ SH ₃ O (604.68) calculated: C 60.6 H 4.8 N 4.6 S 10.4 % found: 60.65 4, 6 4.4 9.6%, R _f 0.35 (vf), 0.55 (s), 0.75 (f) (system B, 0.2 mg load), 0.9 (system C, 0 , 2 mg load), 0.4 (s), 0.65 (f), 0.75 (mf) (system D, 0.1 mg load), 0.0 (Merck plates, developed with ethyl acetate).

Example 9

Reaction between 3-chloromethyl-> 7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid and thallium -! - salts of ß-keto compounds

A. Salt of ethyl benzoylacetate

The thallium I salt of ethylbenzoylacetate (255 mg, 0.625 mmol, 2.5 equivalents) was added to a stirred solution of 3-chloromethyl-7β- (2-thienylacetamido) -ceph-3-em at a temperature of 25 ° C -4-carboxylic acid (107 mg, 0.27 mmol) in dry tetrahydrofuran (1 ml). A precipitate formed immediately and the suspension was brown after 5 minutes. After 4 hours, the chromatography (system C) showed a spot of medium intensity with an R _f value of 0.65, that of 3- (2 ^ -Benzoyl-2 Ϊ -ethoxycarbonylethyl) -7ß- (2-thienylacetamido) -ceph -3-em-4-carboxylic acid corresponded, and a heavy spot at R _f 0.0.

109843/1954

21H541

B. acetylacetone salt

The experiment was similar to that of A and gave a product which, when chromatographed in system C, gave a spot of medium intensity at an R _f value of 0.6, which corresponds to the 3- (2 ^ - acetyl-3-oxobutyl) - 7β- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid and had an intense spot at R _f 0.0.

C. Ethyl acetoacetate salt

The experiment was similar to that of A and gave a product which, when chromatographed in system C, gave a spot of medium intensity with an R _f value of 0.8, which corresponds to 3- (2fc xthoxycarbonyl-3-oxobutyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid and had an intense spot at R _f 0.0.

Example 10

a) Diphenylmethyl-3- (3-oxobutyl) -7β- (2-thienvlacetamido) -ceph-3-em-4-carboxylic acid

A suspension of 5.47 g of 3- (2i -diphenylmethoxycarbonyl-3-oxobutyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid (9 mmol) in 6 ml of anisole was mixed with 24 ml of trifluoroacetic acid Shaken 22 C until a solution was obtained after 7 minutes. The light brown product was evaporated at 22 ° C./2 mm Hg for 15 minutes in a rotary evaporator, dissolved in sodium bicarbonate (pn value 8) and washed with ethyl acetate. The aqueous layer was acidified to a _pH value of 2 and extracted with ethyl acetate, which was washed, (g 3.7, 100%) and dried to form a foam was evaporated. The crude foam was dissolved in 45 ml of tetrahydrofuran and treated at 20 to 23 ° C. with a solution of 3 g of diphenyldiazomethane in 30 ml of petroleum ether. The purple solution was rotary evaporated, dissolved in ethyl acetate and added to a large volume of petroleum ether to give a light yellow powder of the crude ester (3.8 g,

109843/1954

-.52 - '

72%), R _f 0.6 (mf), 0.35 (f), 0.3 (s) and 0.0 (m) (system E). This solid was chromatographed over silica gel (180 g), a mixture of benzene / ethyl acetate (4/1) eluting the title compound (1.85 g, 36 % of the acid), which crystallized from ethyl acetate in the form of prisms, F = 157 until

159 ° C (dec.), [Α] + 4.3 ° (tetrahydrofuran), UV spectrum λ

m Q.2C *

235 nm (fc 13500), inflection at 260 nm (^ 7300), IR spectrum ι / (CHBr-) 3420 (NH), 1780 (ß-lactam), 1715 (CO "R and COMe)

ΓΠα.Χ # -J si c-

and 1680 and 1510 cm (CONH), V 3310 (NH), 1755 (ß-lactam),

' Max. ' . '

1715 and 1705 (CO ₂ R and COMe) and 1670 and 1525 cm (CONH), NMR spectrum Γ (100 MHz; CDCl ₃ , CDCl ₃ with D ₃ O) 2.6 (10 protons; Ph ₂ ), 2.7 to 3.1 (m; 2-thienyl), 3.14 (1-proton s; CHPh ₃ ),

3.6 (1-proton d, J 9 Hz; CONH), 4.25 (1-proton (j, J 5 Hz and

9 Hz; 'C ₇ -H), 5.1 (1-proton d, J 5 Hz; C ₅ -H), 6.18 (2-proton s; CH ₂ -CO), 6.56 and 6.82 (2 -Proton AB-q, J 18 Hz; C ₂ -CH ₂ ), 7.2 to

7.7 (4-proton m, possibly A ₂ B ₂ system; C ₃ -CH ₂ CH ₂ CO) and 7.98 (3-proton s; COMe); deuteration removed the doublet at 3.6 t and gave the quartet at 4.25 τ a doublet, J 5 Hz. C ₃₀ H ₂₈ N ₃ O ₅ S ₂ (560.67):

calculated: C 64.3 H 5.05 N5.0 S 11.4% found: 64.1.5.1 4.6 11.8%, R _f 0.3 (s) (system E, 0 ,, l mg load).

b) 3- (3-Oxobutyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid

1.407 g of diphenylmethyl 3- (3-oxobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate were dissolved in 2 ml of anisole and 8 ml of trifluoroacetic acid. After 4 minutes the solution was evaporated at 2 mm Hg pressure. Ethyl acetate was added and evaporated and the resin was partitioned between ethyl acetate and water at p "8.0. The aqueous layer was acidified to a pn value of 2 and extracted with ethyl acetate, which was then washed, dried and evaporated to give a foam (887 mg, 90%). The foam was crystallized from 2 ml of ethyl acetate in the form of prisms of the title compound (400 mg, -41%). The filtrate was

109843/1964

ORIGINAL INSPECTED

added to petroleum ether to give a second batch (450 mg, 46%). The crystals had an F = -113 to 117 ° (decomp.). [aJ _D + 45 ° (CHCl ₃ ), UV spectrum \ _max (p _H 6 0, lm-phosphate) 236 nmi (& 12500), inflection at 260 nm (8 7900), IR spectrum V

ΓΠΟ.Λ ·

3500 (H ₂ O), 3295 (NH), 1768 (β-lactam), 1720 and 2600 (CO ₂ H), 1696 (possibly COMe) and 1667 and 1522 cm " ¹ (CONH), NMR spectrum τ (100 MHz 5 D ₃ O with NaHCO ₃ ) -2.69 and "3.01 (3-proton m; 2-thienyl), 4.50 (1-proton d, J 5 Hz; C ₇ -H), 5, 00 (1-proton dm J 5 Hz; C ₅ -H), 6.15 (2-proton s; CH ₂ CO) 6.51 and 6.90 (2-proton AB-q, J 19 Hz; C ₃ -CH ₂ ), 7.2 to 7.7 (4-proton m; C ₃ -CH ₂ CH ₂ CO) and 7.81 (3-proton s; COMe); there were also signals at 5.86, 7.74 and 8.75, which correspond to 0.2 mol of ethyl acetate. C ₁₇ H ₁₈ N ₃ O ₅ SO, 5 H ₂ O-0.2 EtOAc:
calculated: C 50.6 H4.7 N6.6 S 15.3% found: 50.5 4.8 6.1 15.3%, R _f 0.6 (system C, 0.2 mg loading), 0 , 18 (system B, 0.2 mg load), 0.25 (vf), 0.3 (vs) (system D, 0.1 mg load).

Example 11

a) Diphenylmethyl 3- (2 \ - methyl-3-oxopentyl ) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylate, second isomer

Diphenylmethyl-3-iodomethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (8g, 12.8mmol) was treated with N- (pent-2-enyl) -pyrrolidine (8 ml, approx , 4 equivalents) in 160 ml of dry benzene according to the procedure of Example 1 a, so that after purification by chromatography over Merck silica gel (0.05 to 0.2 mm) a foam (1.079 g, 14.4%) received. This foam was dissolved in 10 ml of ethanol and the solution was cooled so that the diphenylmethyl _{ester T} first isomer (310 mg, 4 %) was obtained in the form of colorless needles, mp = 145 ° C. (decomp.), [Tt] ^ ³ + 20 ° (£ 1.06, CHCl ₃ ), UV spectrum X _max inflexion (etOH) 260 nm (& 6800). [The IR and NMR spectra of this material were identical to those described in Example Ia) '. The remaining ethanol solution was evaporated to a foam (700 mg), which was dissolved in ethyl acetate and then added dropwise Solution in petroleum ether

109843/1964

- 54 - 21H541

so that the second I some re »was obtained as a white amorphous solid (495 mg, 6.6 %) . A pure sample of the second isomer was obtained by combining the amorphous products obtained from a variety of such reactions and purifying by further chromatography over Merck silica gel (0.05-0.2 mm) with benzene / ethyl acetate (iio / l) obtained as an eluent. The foam obtained by evaporating these fractions with an R _f value of 0.4 (thin-layer chromatography, silica gel ^GF ₂ 54> developed with benzene / ethyl acetate = 5/1) 'was dissolved in ethyl acetate and the solution was poured into petroleum ether so that an amorphous solid, F = 56 to 70 ° C (decomp.) was obtained, [α. | _β -3.4 ° (c 1.1, CHCl ₃ ), UV spectrum λ _{Ιη £ 1θχ ± οη} (EtOH) 260 nm (£ 6900), UV spectrum \> (CHBr.,) 3380 (NH), 1770 (ß-lactam),

ΓΠ3.Χ · j ^

1715 (CO ₂ R), 1700 (C = O) and 1670 and 1505 cm (-CONH); the NMR spectrum (CDCl ₃ ) showed that this material was essentially the second isomer with signals at t 2.69 [Ph ^ CH-J, 2.73 to 3.08 (3-proton multiplet, 2-thienyl), 3.09 (Ph ₂ CiO, 3.32 (-NH, doublet, J 9 Hz), 4.29 (C ₇ -H, double doublet. J 5 and 9 Hz), 5.10 (Cg-H, doublet , J 5 Hz), 6.18 (2-thienylmethyl), 6.73 (C ₃ -H, two proton singlet), 7.2 to 7.9 (-CH ₂ CH _- (CH ₃ ) COCJi ₂ CH _3, 5-proton envelope), 8.79 (CHCH _3, DubleTtt, J 6 Hz) and 9.02 (-CH ₂ CH _3, triplet, J 7Hz) with a small amount (10 to 15%) of the first isomer was indicated by signals at 6.68 and 6.93 (C ₃ -H, AB-quartet, J

18 Hz) is characterized; ^C ₃₂ ^H 32 ^N 2 ° 5 ^S 2 ⁽⁵⁸⁸ » ^7): calculated: C 65.3 H 5.5 N 4.8 S 10.9% found: 65.1 5.4 4.5 11.2% .

b) Diphenylmethyl 7β-amino-3- (2 f -methyl-3-oxopentyl) -ceph-3-em-4-carboxylate, hydrogen p-toluenesulfonate, second isomer

The product of part a (8.0 g, 13.6 mmol) was mixed with 12.8 ml (160.8 mmol) of pyridine and 8.52 g (40.6 mmol) of phosphorus pentachloride in 320 ml of dry methylene chloride, followed by 140 ml Methanol, treated by the procedure of Example 2a to give an oily resin (10.69 g). A solution of the resin in 30 ml of ethyl acetate and 60 ml of ether was made

3/1854

(dec.) was treated with 2.8 g (14.7 mmol) of p-toluenesulfonic acid hydrate in 30 ml of ethyl acetate, so that colorless needles of p-toluenesulfonate (3.63 g, 42%) of F = 155 to 160 ⁰ C. , [<x] p ³ -4.7 ° (c 0.9, CHCl ₃ ), UV spectrum X ^ _3x (EtOH) 257 nm (6-6400), IR spectrum V (Nujol) 2650 (NH +) , 1788 (β-lactarn), 1722 (CO2R), 1710 (C = O) and 1010 cm (SO ₃ "-), NMR spectrum τ (DHSO-d ⁶ ) 2.48 and 2.91 (^ CH ₃ C ₆ H ₄ SO ₃ -), 2.61 (Ph ₃ CH-), 3.08 (Ph ₂ CI ^ -), 4.76 (C ₇ -H, double doublet), 4.87 (C ₅ - H, doublet, J 5 Hz), 6.44 (C ₂ -H, broad singlet), 7.2 to 7.6 (-CH ₀ CH (CH ₃ ) COCIi ₂ CH ₃ , 5-proton envelope), 7 , 69 (P-CH ₃ CgH ₄ -), 9.12 ~~ (triplet superimposed over a doublet, CHClH ₃ and -CH ₂ CH ₃ ) Jc ₃₃ H ₃₆ N ₂ O ₇ S ₂ (636.8):
calculated: C 62.2 H 5.7 N 4.4 S 10.1% found: 62.5 5.7 4.0 9.9%]:

R 0.3 (thin layer chromatography, silica gel ^GF 254 + 366 ~ ^Pla ^ "" th, developed with benzene / ethyl acetate = 2/1). [The NMR spectrum did not allow a quantitative distinction between the two isomers. J.

c) Diphenyl 3- (2 \ -methyl-3-oxopentyl) -7β- [D-2- (tert-butoxycarbonylamino) -2 -phenyl-cetamido)] -ceph-3-em-4-carboxylate, first and second isomer.

A solution of N- (tert-butoxycarbonyl) -D-phenylglycine (1.86 g, 7.72 mmol) in 20 ml of dry tetrahydrofuran was at -10 ° C with 0.96 ml (7.72 mmol) of triethylamine, followed by 1.02 ml (7.72 mmol) isobutyl chloroformate in 10 ml of dry tetrahydrofuran, while keeping the temperature below -6 ⁰ C. The suspension was stirred for 30 minutes while allowing the temperature to approximately 20 ⁰ C rise. The precipitated triethylamine hydrochloride was filtered off and the filtrate was added to a solution of diphenylmethyl-7β-amino-3- (2t-methyl-3-oxopentyl) -ceph-3-em-4-carboxylate-hydrogen-p in the course of 5 minutes -toluenesulfonate, second isomer (3.198 g, 5.03 mmol) in 20 ml of acetonitrile and 10 ml of Ν, Ν-dimethylacetamide. After stirring for 1 hour, the solvents were removed in vacuo, and a solution of the residue in 100 ml of ethyl acetate was washed with saturated sodium bicarbonate / water for 30 minutes.

109843/1954

21H541

ser (l / l) (100 ml) stirred, and the organic phase was separated, washed with 100 ml of saturated sodium bicarbonate, 2 × 50 ml of water and 50 ml of brine, dried over magnesium sulfate and evaporated to a foam (3.6 g) . This material was purified by chromatography on Merck silica gel (0.05-0.2 mm) (80 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions, R, - 0.4 (thin layer chromatography, silica gel GFpt-. ..,., -, developed with benzene / ethyl acetate = 5/1) ■ resulted in a foam (2.65 g, 76%) -, that from ethyl acetate: petroleum ether. .Precipitated to give 2.13 g (61 %) of a white solid. This material was dissolved in 20 ml of ethanol, the solution was cooled to 0 ^{° C. and the white precipitate of the} title ester was filtered off (240 mg, 6.8 %). [This sample was a mixture of approximately 70% of the first and 30% of the second isomer.] The filtrate was evaporated to a foam which was precipitated from ethyl acetate / petroleum ether to give a white amorphous solid (1.4 g , 40%), F = 110 to 113 ° C (decomp.), [Aj ^ ³ -18.8 ° (c 0.81, CHCl ₃ ), UV spectrum λ (EtOH) 259 nm (£ 7300), IR spectrum>>"(CHBr-.),

ΓΠ3Χ · "ΓΠαΧ · ο

3400 (NH), 1782 (β-lactam), ca.1710 (CHCO ₂ R, CO ₃ R, C = O) and 1690 and 1520 cm " ¹ (CONH); the NMR spectrum (CDCl ₃ ) showed that this material was a mixture of approximately 60% of the first and 40 % of the second isomer with signals at 2.67 (Ph ₂ CH and PhCH), 3.09 (Ph ₃ CH), 4.28 (C ₇ -H, double doublet, J 5 ~ and 9 Hz), 4.36 (tBuOCONH-, doublet, J 8 Hz), 4.80 (PhCH, doublet J 8 Hz), 5.13 (C ₅ -H, duhlet, J 5 Hz), 6.66 and 6.89 (C ₂ -H, AB-quartet, J 18 Hz), Ι, ά to 7.9 (-CH ₂ CH (Ch ₃ ) COCH ₂ CH ₃ 5-proton envelope) , 8.59 [(CH ₃ J ₃ C-], 9.01 (CH ₃ CH ₂ -, triplet, J 7 Hz) and 9.09 (CH ₃ CH, doublet, J 6 Hz) for the second isomer and at 6.63 and 6.91 (C ₃ -H, AB-quartet, J 18 Hz) for the first isomer. [ C ₃ ^ H ₄₃ N ₃ O ₇ S (697.8): calculated: C 6 7, 1 H 6.2 N 6.0 S 4.6% found: 66.4 6.2 5.8 4.6%]

109843 / 1.9 54

© WÖINAi »INSPECTED

21 USA 1

d) 7β- (D-2-amino-2-phenyl acetamido) -3- (2 F -methyl-3-oxopentyl) ■ ceph ~ 3-em-4-ca rb onic acid, trifluoroacetic acid salt;, second isome / eres (with first isomer)

The ester of part c (1.31 g, 1.88 mmol.) Was added with 2.5 ml

Anisole and 10 ml of trifluoroacetic acid, and after 5 minutes the reagents were separated at 30 ° C./2 mm Hg. The residue was suspended in 50 ml of ether and the suspension was shaken with 50 ml of water containing 3 drops of trifluoroacetic acid. The aqueous phase was separated off, washed with 2 × 50 ml of ether and 2 × 50 ml of ethyl acetate and freeze-dried to give a white crystalline solid (800 mg, 78%), mp = 110 to 143 ° C. (decomp.), [Aj ^ ⁶ + 74.1 ° (c 1.11, acetone / water _β l / l) UV spectrum \ _m _ 262.5 nm (6 8400), IR spectrum • _{M v}

lTlcLX # ^ UlOiC *

(Nujol) 1756 (β-lactam) and 1680 cm (C = O and CF ₃ CO ₂ -); the NMR spectrum showed that this material was a mixture of the second and first isomers (ca. 60i40) with signals at -C (D ₂ O) 2.46 (C ₆ H ₅ -), 4.40 (C ₇ -H, doublet), 4.96 (C ₅ -H, doublet, J 4.5 Hz), 4.75 (C ₆ H ₅ CjH (NH ₃ ) -), 6.60 and 6.96 (C ₃ -H, AB-Quartet, J 18 Hz), 6.90 to. 7.55 - (-CH ₃ CH (CH ₃ ) COCH ₂ CH ₃ , 5-proton envelope), 8.96 (CH ₃ CH ₂ - overlaid on CHCH_ ₃ ) for the second isomer and at 4.38 (C ₇ -H, doublet), 4.94 (C ^ -H ₁ doublet »J 4.5 Hz) and 6.58 and 6.90 ^ (AB-quartet, C ^ -H, J. 18 Hz) for the first . isomers [C ₂₃ N ₂₆ F ₃ N ₃ O ₇ S (545.3). Calculated: C 50.6 H 4.8 F 10.45 N 7.7 S 5.9 % found 50.9. 5.1 7.65 7.9 6.2 Ιέ; these figures suggest that this material contained free acid »] The electrophoretic and chromatographic properties of this material were compared with those of an authentic sample,

Example 12

a) Diphenylmethyl-3-benzoylethyl-7ß- (2-'thienvlacetamido) -ceph-3-em-4-carboxylate

A solution of 3-iodomethyl-Diphenylfnethyl-7ß- (2-thienylacetamidö) ceph-3-em-4-carboxylate (12.6 g, 20 mMöl) in dry methylene chloride (250 ml) was stirred vigorously at 25 ⁰ C and

1 0 9 Ö 4 3/1 9 S 4

21U541

treated in the course of 1 minute with 10 ml. (approx. 4 equivalents) 1-pyrrolidine styrene. The solution was stirred for a further 15 minutes and then stirred with 200 ml of 1N hydrochloric acid for 5 minutes * The organic layer was separated, washed with 200 ml of saturated sodium bicarbonate solution, 200 ml of water and 200 ml of brine, dried over magnesium sulfate and evaporated to a resin 16.645 g ) This material was purified by chromatography over silica gel (Merck), 0.05 to 0.2 mm (340 g) with benzene / ethyl acetate (lo / l) as the eluent. A combination of similar fractions (according to the thin layer chromatogram) gave 4.88 g of a foam which was crystallized from ethanol to give the title compound in the form of pale yellow crystals (2.816 g, 22.5 %) , R _f 0, 44 (thin layer chromatogram, benzene / ethyl acetate = 5/1), F = 171.5 to 172.5 ° (decomposition, C «] β ³ ~ 29 ° < ^C 07, chloroform) _y

UV spectrum λ (EtOH) 240 nm (£ 25400), inflexion at ^r max * '

260 nm (£ 10500), IR spectrum \>. (CHBr,) 3403 (NH), 1776 (ß-Laetam), 1720 (CO ₂ R), 16 85 (ketone) and 16 79 and 1510 cm (CONH), NMR spectrum ¹ S (CDCU) 2.15 and 2.62 (0.H ₁ -CO), 4.21 (C ₇ ^ -H, double doublet, J 4.5 and 9 Hz), 5, o5 (C ^ xH), doublet, J 4.5 He ), 6.48 and 6.72 {C ₍₂ j-CH ₂ , AB-quartet, J _AB 18 Hz) and 6.7 to 7.5 (unresolved multiplet, -CHgCH ₂ COC ₆ H ₅ ) · [ C ₃₅ H ₃₀ N ₂ O ₅ S ₂ (622.7): Calculated: C 67.5 H 4.85 N 4.5 S 10.3 % found: 67.3 x 4.8 4.3 10.2 %]

b) 3- (2-Benzoylethyl) -7ß- (2-thienylacetamido) -ceph-3-ero-4-carboxylic acid

A solution of diphenyl 3- (2-ben2; oylethyl) -7ß- (2 »thienylacetamido) -ceph-3-em-4-carboxylate- (2.5 g, 4.02 mmol) in 10 ml of trifluoroacetic acid and 2.5 ml of anisole was 5 minutes stirred at 25 ° C. The solvents were separated off in vacuo and the residue was partitioned between ethyl acetate and 2N sodium bicarbonate solution distributed. The acidification of the bicarbonate solution to a p ^ value of 2 the acid gave (1.848 g, 100%) ..

109843/1954

21H5A1

Trituration of this material with ether gave the acid (900 mg, 49%) in the form of a pale yellow solid, F = 94 to 102 ° C (Zprs.), U] ^ ⁶ + 24.1 ° (c 1.1 , Chloroform), UV spectrum λ (Ο, Ιπι-PhosDhatbuffer, p _u 6) 243 nm (£ 21500), ¹ max. 'Rl

Inflection at 260 nm (t 13300), IR spectrum V _max (CHBr ₃ ) 3440 (NH), 1780 (β ~ I.actarn), 1735 (CO ₂ H), 1690 (ketone) and 1680 and 1512 cm " ¹ (CONH), NMR spectrum r (dimethyl sulfoxide - d ⁶ ) 1.95 and 2.35 (C ₆ H ₅ CO), 4.34 (C _(y) -H, double doublet, J 4.5 and 9 Hz), 4.90 (C ^ ₆ JH, doublet, J 4.5 Hz), 6.37 (C ₍₂₎ -CH ₂ , singlet) and 6.70 and 7.28 (unresolved multiplets, - CH-CH-COC ^ -H, -), Rj-0.53 (Whatman # 1 paper, buffered

—Ve —— et OJI

a p n value of 5, downward development at 37 ° C. with an upper phase of n-butanol / ethyl acetate / O, 1 m-sodium acetate = 1/8/8) and R _f 0.46 (Whatman paper No. 1, buffered to a p "value of 6, downward development with an upper phase of n-butanol / ethanol / water = 4 / l / 5, in equilibrium with the

lower phase at 25 ° C) [ ^c ₂ 2 ^H 2O ^N 2 ° 5 ^S 2 ⁽⁴⁵⁶ > ^5): calculated: C 57.9 H4.4 N6, l S 14.05%

found: 5 7.25 4.65 5.5 13.2%. This sample ent

held ether (0.2 mol as indicated by the NMR spectrum.

Example 13

Diphenylmethyl 3- (2 ^ -benzoxlpropyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylate

A solution of diphenylmethyl-3-iodomethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (1.26 g, 2 mmol) in 25 ml of dry methylene chloride was stirred at 25 ° C. and allowed to continue 1 minute with l-pyrrolidono-2-methylstyrene (1.0 ml, approx. 4 equivalents). The solution was stirred for 10 minutes, 25 ml of 1N hydrochloric acid were added and the mixture was stirred for a further 5 minutes. The organic layer was separated with Washed 25 ml of saturated sodium bicarbonate solution, 25 ml of water and 25 ml of brine, dried over magnesium sulfate and evaporated to a foam (1.41 g). This material was chromatographed on 0.05-0.2 mm silica gel (Merck) (28 g) with benzene ethyl acetate = lo / l as elution

109843/1964

21U5A1

- -60 -

medium cleaned. The combination of the fractions, R _f 0.54 (thin layer chromatography, silica gel GF -, -, - plates, benzene / ethyl acetate (5/1) as eluent) gave a white amorphous solid (62 mg, 4.9 %), F = 75 to 88 ° C (dec.), [A] p ³ -1.9 ° (c 1.1, CHCl ₃ ) UV spectrum \ _max (EtOH) 238.5 nm (.t 19300), A. _Inflexion 260 nm ( t 9200), IL spectrum (CHBr,) 3400 (NH), 1780 (ß-lactam), 1720 (CO, R), 1685 (COPh)

-1
and 1580 and 1515 cm (CONH); The NMR spectrum showed that this material was a mixture of isomers (about l / l) of the Ti tel Getting Connected was with signals T _(CDCl3) 2.18 and 2,60-

(COPh), 4.18 and 4.27 (C ₇ -H, two double doublets, J 5 and 9Hz), 5.15 and 5.22 (C ₅ -H, two doublets, J 5 Hz), 6, 3 to 7.4 (unresolved 5-proton multiplet, C ₃ -CH ₃ overlaid on -CH ₂ CH-) and 8.86 and 8 ₅ 98 [(CH ₃ -), two doublets, J 7 Hz].

Example 14

^a ^ Piphenylmethyl 3- ( 2ν -formyl-n-propyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate

A solution of diphenylmethyl-3-iodomethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (6.30 g, 10 mmol) in dry methylene chloride (130 ml) was stirred at 23 ° C and treated with 1-di-n-butylaminoprop-1-ene (6.75 mL, about 8 equivalents) over 1 minute. The solution was stirred for a further 30 minutes and washed successively with 1 × 80 ml and 1 × 50 ml of IN hydrochloric acid, 50 ml of saturated sodium bicarbonate solution, 50 ml of water and 50 ml of brine, dried over magnesium sulfate and given an oily solid (8.2 g ) evaporated. This material was purified by chromatography on 0.05-0.2 mm silica gel (Merck) (160 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions with an R _f value of 0.35 (thin layer chromatogram, silica gel GF ₂₅₄ plates, benzene / ethyl acetate = 5/1 as eluant) gave an oily solid (2.078 g) which was dissolved in 2 ml of ethyl acetate , and after cooling the solution gave the title compound (163 mg, 3%) as colorless needles, F = 183 to 185 ° C (decomp.), [a] p ³ + 7 °. (c 0.57, CHCl ₃ ) , UV spectrum

. 109843/1954

21U541

(EtOH), 260 run (£ 6800), ^ _Inflexlon 235 nm (S 13600), IR spectrum V (CHBr ,,) 3420 (NH), 2730 (CHO), 1781 (β-lactam), 1720 (CO ₂ R and CHO) and 1690 and 1512 cm (CONH); the NMR spectrum (CDCI,.) showed that this material was a mixture (approx. 3: 2) of diastereoisomers with signals at T 0.48 ■ (CHO), 3.05 (CHPh ₂ ), 4.21 ( C ₇ -H, double doublet, J 5 and 9 Hz), 5.03 (C ₆ -H, doublet, J 5 Hz), 6.55 and 6.83 (C ₃ -H, AB-quartet, J 18 Hz), 7.2 to 7.6 (3-proton envelope, -CHgCH-) and 9.03 (CH _-3 -CH-, doublet, J 7 Hz) for the main component and at 0.52 (CHO), 6.67 (C ₃ -H) and 8.95 (CIi ₃ -CH-, doublet, J 7 Hz) for the smaller component. The NMR spectrum and microanalysis showed that this material was contaminated with water and ethyl acetate. [C ₃₀ Hp ₈ N ₂ O ₅ S ₂ (560.7): Calculated: C 64.3 H4.9 N5.0 S 11.4% found: 64.65 5.3 4.35 10.8 %]

b) 3- (2 ^ -Formyl-n-propyl) -7ß- (2-thienvlacetamidec? -ceph-3-em-4-carboxylic acid

Diphenylmethyl 3- (2-formylpropyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylate (approx. 3: 2 mixture of the diastereoisomers, 123mg, 0.22mmol) was 0.15 ml of anisole and 0.5 ml of trifluoroacetic acid and after 5 minutes the reagents were evaporated at 30 ° C./2 mm Hg. The residue was partitioned between 20 ml of ethyl acetate and 10 ml of saturated sodium bicarbonate solution / water (l / l). The acidification of the bicarbonate solution. a _pH value of 2 gave the acid (78 mg, 90%) as an oil, from which a solution was poured into ethyl acetate in petroleum ether, so that it is in the form (mg 47, 54%), a purer sample of amorphous solid obtained, UV spectrum λ (0.1m phosphate buffer, p _H 6) 236 nm ( t 13000), ^λ _Ιηί1θΧ ^ _οη 260 nm ( t 7850), IR spectrum V (CHBr,) 3420 (NH ) 1782

ΓΠ3Χ j ^ i

(β-lactam), 1724 (CO ₃ H and CHO) and 1688, cm (CONH); the NMR spectrum (DMSO-d _g ) indicated that this material was a mixture (ca. 3: 2) of the diastereoisomers with signals at? TO, 36 (-CHO), 0.87 (NH, doublet, J 9 Hz), 2.60 and 3.02 (thien-2-yl) 4 ^ 8 (C ₇ -H., Double doublet, J 5 and 9 Hz), 4.89 (C ₆ -H, doublet, J. 5 Hz), 6.22. (thien-2-yl-CH_ ₂ ) 6.35 and

109843/1954

21H541

6.59 (C ₂ -H, AB-quartet, J 18 Hz), 7.0 Ms 7.5 (-CHpCH-, 3-proton envelope) and 9.00 (CH ^ -CH-, doublet, - J 7 Hz) for one isomer and at 4.32 (C ₇ -H-, double doublet, J 5

and 9 Hz), 4.90 (Cg-H, doublet, J 5 Hz), "6.49 (CH, singlet) and 8.94 (CH ₃ CH-, doublet, J 7 Hz) for the other isomer. R _f 0.15 (Whatman paper No. 1, buffered on p "5, downward development at 37 ° C. with an upper phase of n-butanol / ethyl acetate / o, lm-sodium acetate = 1/8 / S) and R _f 0 , 46 (Whatman paper No. 1, buffered on p "6, downward development with an upper phase of n-butanol / ethanol / water = 4 / l / 5, in equilibrium with the lower phase, at 25 ° C.).

Example 15

a) Diphenylmethyl 3- (2-formylisobutyl) -7β- (2- thienylacetamido) -ceph-3-em-4-carboxylate

A solution of diphenyln; ethyl-3-3odmethyl-7ß ~ (2-thienylacetamido) -ceph-3-em-4-carboxylate (12 _F 6 g, 20 mmol) in 250 ml of dry methylene chloride was stirred at 23 ° C and with Treated 10 ml (approx. 4 equivalents) of 1-pyrrolidinoisobutene. The solution was stirred for 15 minutes, 200 ml of 1N hydrochloric acid were added over the course of 5 minutes, the organic phase was then separated off and washed successively with 200 ml of saturated sodium bicarbonate solution, 2OO ml of water and 200 ml of brine, treated with approx. Filtered through a bed of kieselguhr, dried over magnesium sulfate and evaporated to a foam (10.98 g). This material was purified by chromatography over 0.05-0.2 mm silica gel (Merck) (400 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions with an R. _p value of 0.48 (thin-layer chromatography, silica gel GFp ₁ ... ", benzene / ethyl acetate = 5/1) gave the ester (2.49 g, 21.7 %) in the form of a foam. A purer sample was obtained by crystallizing this foam from ethyl acetate / ether (1/1) (25 ml) in the form of colorless needles (965 mg, 8.4%). F = 161 to 165 ° C (decomp.), [A] ^ ³ + 5.7 ° Cc 1.0, CHCl ₃ ), UV spectrum *: _{Inflex #} (EtOH), 260 nra C t 680O), * _Inflex 235 nm

109843 / 19Bt

21U5A1

( ε 14000), IR spectrum V (CHBr-.) 3415 (NH), 2710 (CHO), 1780 (ß-lactam), 1722 (CO ₂ R and CHO) and 1680 and 1510 cm (COIv ¹ H), NMR Spectrum T (CDCl ₃ ) 0.64 (CHp), 4.28 (C ₇ -H, double doublet, J 5 and 9 Ha), 5.05 (Cg-H, doublet, J 5 Hz), 6 , 84 and 7.04 (C ₃ -H, AB-quartet, J 17 Hz), 6.99 and 7.53 (C ₃ -CH_ ₂ -, AB-quartet, J 14 Hz) and 9.11 [( CH ₃ ^ C-)]. t ^C 31 ^H 3O ^N 2 ° 5 ^S 2 ⁽⁵⁷⁴ > ^7): calculated: C 64.8 H 5, TN 4.9 S 11.1%

found: 64.5 5.3 4.7 .11.2%]

b) 3- (2-Formylisobutyl) -7β- (2-thienylacetamido) -ceph-3-em ~ 4 ~ carboxylic acid

Diphenymethyl-3- (2-formylisobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (880 mg, 1.53 mmol) was treated with 1 ml of anisole and 4 ml of trifluoroacetic acid, and after 4 minutes the reagents were evaporated at 30 ° C / 2 mm Hg. The residue was partitioned between ethyl acetate (30 ml) and saturated sodium bicarbonate solution / water = l / l (30 ml). The acidification of the bicarbonate layer to a p _H value of 2 gave the acid (380 mg, 61%) as an oily solid. This material was dissolved in ethyl acetate and the solution poured into petroleum ether to give a purer sample as an amorphous solid (280 mg, 45%), mp 73-105 ° C (Zefs.), [A] ^ ³ + 8.3.1 ° (c 0.83, tetrahydrofuran), UV spectrum λ (0, lm ~ Phosphatpüfferj p "6) 235 nm (£ 136ΟΟ), λ _{Σ fl} 260 nm (fc 8300), IR spectrum V _max (CHBr ₃ ) 3430 (NH), 1784 (ß-lactam), 1728 (CO ₂ R and CHO) and 1690 and 1615 on " ¹ (CONH), NMR spectrum T (D ₂ O, NaHCO ₃ ) 0, 44 (CHO), 2.63 and 2.95 (thien-2-yl), 4.44 (C ₇ -H, doublet, J 5 Hz), 4.92 (C ₆ -H, doublet, J. 5 Hz), 6.10 (thien-2-yl-CH_ ₂ -), 6.59 and 6.94 (C ₂ -H, AB-quartet, J 18 Hz), 6.88 and 7.60 (C ₃ -Oi ₂ -, AB-quartet, J 14 Hz) and 8.90 and 8.92 [(CH ₃ ) ₂ ~, two singlets]. [C ₁₈ H ₂₀ N ₂ O ₅ S ₂ (408.5): calculated: C 52.9 H 4.9 N 6.85 S 15.7%

found: 53.1 5, O 6.6 15.2%]

R _f O.73 (Whatman paper No. 1, _{buffered on p H} 5, downward development at 37 C with an upper phase of n-butanol / ethyl acetate / o, Im sodium acetate = ΐ / δ / 8) and R _f 0 , 38 (Whatman paper no.!,. Buffered on p _u 6, downward development with an upper

^H 109843/1954

21U541

- "64 -

Phase of n-butanol / ethanol / water = 4 / l / 5, in equilibrium with the lower phase, at 25 C).

Example 16

^a > Diphenylmethyl-3 - ( 2 % -formyl-2 ^ -phenylethyl) -7ß- (2-thienyl- acetamido) -ceph-3-em-4-carboxylate

A solution of 1.26. g (2 mmol) of DIphenylmethyl-3-iodomethyl-7β- (2-thienylacetamido) -ceph-3 ~ em-4 -carboxylate in 30 ml of dry methylene chloride was treated with 1 ml (about 4 equivalents) of N-styrylpyrrolidine, and the resulting solution was stirred at 23 ° C. for 30 minutes. Then 20 ml of n-hydrochloric acid were added and stirring was continued for a further 5 minutes, then the organic phase was separated, washed with 20 ml of saturated sodium bicarbonate solution, 20 ml of water and 20 ml of brine, dried over magnesium sulphate and added at 1.5 l g of a foam evaporated. This material was purified by chromatography over 0.05 to 0.2 mm silica gel (Merck) (100 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions with an R _f 0.53 (thin layer chromatography, silica gel ^GF 254 + 366 »developed with benzene / ethyl acetate = δ / 1) gave a foam (600 mg) which was precipitated from ethyl acetate / petroleum ether so that the title compound (530 mg, 42.6%) of an amorphous solid in the form of, F = 72-88 ⁰ C (dec.), [a] ^ ³ -38.1 ° (c 1.05, CHCl _3), UV -Spectrum ^X Inflex ^(Et0H) 260 nm (£ 7700), X _Inflex 235 nm (& 14400), IR spectrum V _mB ^ _m (CHBr ₃ ) 341O (NH), 272o (CHO), 1774 (ß-lactam) , 1716 (CO ₂ R, CHO) and 1676 and 1500 cm " ¹ (CONH); the NMR spectrum (CDCl ₃ ) showed that this material was a mixture (approx. 2: 1) of the diastereoisomers with signals at τ 0 , 40 (CHO), 4.23 (CH, double doublet, J 5 and 9 Hz), 5.18 (C ₆ -H, doublet, J 5 Hz)., 6.78 and 7.48 (C ₂ - H, AB quartet, J 18 Hz) and 6.6 to 7.0 and 7.1 to 7.5 (-CH ₂ CH-, two unresolved multiplets) for "· ■ the main component and at τ O, 59 (COH), 5.19 (Cg-H, doublet, J 5 Hz) and 6.69 and 7.45 (C ₂ -H, AB-quartet, J 18 Hz) for the secondary component. [ ^C 35 ^H 30 ^N ₂ ° 5 ^S 2 ^ ⁶² 2.7):. . calculated: C 67.5 H 5.0 N 4.5 S 10.3 %

r S 95 I? '' * ^]

21U541

This material was, as the NMR spectrum showed, with petroleum ether contaminated.

^b ^ 3 ~ (2j ~ Formvl-2 y ~ phenyläthvl) ~ 7ß- (2-thienvlace tamido) -ceph- 3-em-4-carboxylic acid

Diphenylmethyl (3- (2j ^: -formyl-2 l-phenylethyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylate (420 mg, 0.674 mmoles) was mixed with 0.25 ml of anisole and 2 ml of trifluoroacetic acid, and after 4 minutes the reagents were evaporated at 30 c / 2 mm Hg. The residue was partitioned between 20 ml of ethyl acetate and 20 ml of saturated sodium bicarbonate solution / water (l / l). Acidification of the bicarbonate solution on a p "- A value of 2 gave the acid (290 mg, 94%) as an oil. A solution of this oil in ethyl acetate was poured into petroleum ether to give a purer sample (205 mg, 67 %) as an amorphous solid. F = 80 to 95 ° C (dec.), [A] ^ ³ + 38.2 ° (c 0.85, tetrahydrofuran), UV spectrum λ (0, Im-phosphate buffer, p _H 6) 236 nm (& 13 700), * _Inflex 262.5 nm ( t 8450), IR spectrum V (CHBr ,,) 3500 (H ,, 0), 3420 (NH), 2710 (CHO), 1770 (ß-lactam), 1720 ( CO ₂ H), 1710 (CHO) and 1690 and 1510 cm (CONH); the NMR spectrum (DMSO-dg) showed that this material was a mixture ng (approx. 2: 1) the diastereoisomer was with signals at τ 0.31 (CHO), 0.94 (NH), 2.61 (Ph), 2.62 and 3.02 (thien-2-yl), 4.41 (C ₇ -H, double doublet, J 5 and 9 Hz), 5.03 and 5.06 (C ₆ -H, two doublets, J 5 Hz), 6.24 (thien-2-yl-CH_ ₂ ) and 6.2 to 7.1 (C ₂ -H and -Cli ₂ CH-, 5-proton envelope). [C ₂₂ H ₂₀ N ₂ O ₅ S ₂ -H ₂ O (470.0):

calculated: C 56.2 H 4.6 N 5.95 S 13.6% found: 56.2 4.5 5.55 13.7%] R _f 0.61 (Whatman # 1 paper, buffered p "5, downward development at 37 ° C with an upper phase of n-butanol / ethyl acetate / Ojlm-sodium acetate» ΐ / β / 8) and R _f 0.52 (Whatman paper no. 1, _{buffered on p H} 6, Downward development with an upper phase of n-butanol / ethanol / water = 4 / l / 5, in equilibrium with the lower phase, at 25 ° C).

109843/1954

21H541

- if -

Example 17 Diphenylmethyl-3- (2nd ί -ethoxycarbonyl-3 ~ oxo-n ~ bi3tyl) ~ 7ß

a 2 (2-thiienylacetaniido) - ceph-3-em-4- carboxylate and its / | -

Isomer

A solution of 1.26 g (2 mmol) of diphenyl methyl-3-iodomethyl-7β- (2-thienylacetaraido) -ceph-3-em-4-carboxylate in 40 ml of dry methylene chloride was added with 1.48 g (8 mmol ) Treated ethyl 3-pyrrolidinocrotonate in 2O ml of methylene chloride. The solution was stirred for 2 hours at 25 ° C., then 30 ml of 1N hydrochloric acid were added, stirring continued for 5 minutes, the organic phase separated, washed with 30 ml of saturated sodium bicarbonate solution, 30 ml of water and 30 ml of brine, over magnesium sulfate dried and evaporated to a foam (1.265 g). This material was chromatographed on 0.05-0.2 mm silica gel (Merck) (25 g) with benzene / ethyl. acetate = 10 / l purified as eluent. The combination of the fractions with an R ^ value of 0.50 (thin-layer chromatography, silica gel GF _{254 + 36} % plates, developed with benzene / ethyl acetate = 5 / l) resulted in a white amorphous solid (305 mg, 24%), F = 80 to 95 ° C (decomp.), [a] ^ ⁹ + 3.7 ° (c 1.08, CHCl ₃ ), UV spectrum X _Inflex (EtOH) , 260 nm (E / 8000), X _{1 fl} 235 nm (6 13600), IR spectrum Y (CHBr,) 3410 (NH), 1773 "*

til 3.X · «j

(β-lactam), 173Ο (CO ₂ Et), 1716 CCO ₂ CHPh ₂ ), 1706 (COMe) and 1680 and 1500 cm " ¹ (CONH). The NMR spectrum (CDCl ₃ ) showed that this material was essentially the title ester was with signals at 4.20 (C ₇ -H, 1-proton multiplet), 5.07 (C ₅ -H, doublet, J 5 Hz), 5.88, 5, β9 (-CO ₂ CH ₂ CH ₃ , two superimposed quartet, J 7.Hz), 6.2 to 7.2 (CH ₂ CH, superimposed with C ₃ -H ₅ unresolved multiplet), 7.83, 7.94 C-COCH ₃ , two singlets), 8.2 (C = C-Me, about 0.3 proton) and 8.78 (-CO ₂ CH ₂ CH ₃ , triplet, J 7 Hz) with a small amount (about 10%) of diphenylmethyl -3- (2 _i -ethoxycarbonyl-3-oxobutyl) -7ß- (2-thienylacetamido) -ceph-2-em-4-carboxylate (see Example 18), with signals at 4.40 (C ₇ -H, double doublet, J 5 and 9 Hz), 4.88 (C _g -H, doublet, J 5 Hz) and 5.24 (C ₄ -H) [C ₃₃ H ₃₂ N ₂ O ₇ S ₂ -Va H ₃ O (641.7)? Calculated: C 61.8 H 5.2 N 4.4 S 1O ₅ O. %. Found: 61.65 5.0 4.45 9 ₅ 7 %]

109843/1854

21H5A1

Example 18

Diphenylmethyl -3- ( 2f -methoxycarhonyl-3-oxobutyl) -7ß- (2-thienylaceta ^ ido) -ceph ~ 3 ~ em-4 ~ carboxylate

A solution of 1.26 g (2 mmol) of diphenylmethyl-3-iodomethyl-7β «- (2-thienylacetamido) -ceph-3-em-4-carboxylate in 30 ml of dry chloroform was mixed with 1.352 g of C8 mmol) methyl-3 treated pyrrolidonocrotonate in 20 ml of chloroform, and the mixture was refluxed for 1 hour. The solution was then cooled to 25 ° C. and stirred with 25 ml of 1N hydrochloric acid for 10 minutes. The organic layer was separated, washed successively with 25 ml of saturated sodium bicarbonate solution, 25 ml of water and 25 ml of brine, dried over magnesium sulfate and evaporated to a foam (1.275 g). This material was purified by chromatography on 0.05-0.2 mm silica gel (Merck) (25 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions with an R _f value of 0.48 (thin layer chromatography, silica gel ^GF 254 + 366 ~ ^^ ^at * ^en * developed with benzene / ethyl acetate = 5/1) gave an almost white solid. (53O mg, 43%), F x 75 to 87 ° C (dec.), [A] ^ ³ + 41 ° (c 1, O, CHCl ₃ ) J UV spectrum

λ (EtOH), 241 nm (E 16000), IR spectrum >>"(CHBr,) max. ^{J IJT} max. j

3416 (NH), 1775 (β-lactam), 1739 CCO ₂ R), 1700 (COMe) and 1690 and 1515 cm * " ¹ (CONH); the NMR spectrum (CDCl ₃ ) showed that this material was essentially the Ceph-2-em-ester was with signals at "C 4.41 (C ₇ -H ₁ double doublet, J 5 and 9 Hz), 4.90 (C ₆ -H, doublet, J 5 Hz), 5, 26 (C ₄ -H), 6.34 (CO ₂ CH ₃ ), 6.68 and 7.34 (C ₃ -CH _-2 = C, -enol, AB-quartet, J 14 Hz), 7.14 and 7.60 (C ₃ -CH ₂ -CH-, keto-, two superimposed AB-quartets, J 15 Hz) and 7.84, 7.86 (-COCH ₇ , keto- and enol-), together with diphenylmethyl -3- (2-methoxycarbonyl-3-oxobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate (approx. 10%) (see Example 17) with signals at 4.20 (C ₇ -H, double doublet, J 5 and 9Hz), 5, O8 (C ₆ -H, doublet, J 5 Hz) and 6.37 (-CO ₂ CH ₃ ).

₂ O ₇ S ₂ (618.7): calculated * C 62.1 H 4.9 N 4.5 S 10.1%

found: 62.05 5.1 4.15 10.3 %]

109843/1954

21H541

Example 19

Diphenylmethyl-3- ( 2V- tert-butoxycarbonyl-3-oxo-n-butyl ·) - 7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate

Diphenylmethyl-3-iodomethyl-7β- (2-thienylacetamido) -ceph-3-em- 4-carboxylate (1.26 g, 2 nmoles) was dissolved in 20 ml of dry chloroform; then 845 mg (4 mmol) of tert-butyl 3-pyrrolidinocrotonate, melting point 112 to 113 ° C., prepared from pyrrolidine and tert-butyl acetoacetate, in 20 ml of dry chloroform were added and the solution was refluxed for 1 hour. After cooling to 25 ° C., 20 ml of 2N hydrochloric acid were added and the two-phase system was stirred vigorously for 10 minutes. The organic phase was separated, washed successively with 20 ml of saturated sodium bicarbonate solution, 20 ml of water and 20 ml of brine, dried over magnesium sulfate and evaporated to give 1.557 g of a foam. This material was purified by chromatography on 0.05-0.2 mm silica gel (Merck) (60 g) with benzene / ethyl acetate (10/1) as the eluent. The combination of the fractions with an R .. value of 0.5 (thin layer chromatography, silica gel GF _{254 + 366} plates, developed with benzene / ethyl acetate = 5/1) gave a white solid (482 mg, 37%), F = 57 to 67 ° C (decomp.), [A] p + 31.6 ° (c 0.95, tetrahydrofuran), UV spectrum ^λ Ιηί1θχ. ^{(Et0H) 26} ° ^nm ( ^£ 79OO), λ _{Ιη £ 1βχ} 235 nm (& 15100), IR spectrum V “(CHBr-) 3420 (NH), 1780 (ß-lactam), 1730 (CO ₂ R), 1710 (CO ₂ CHPh ₂ and COCH ₃ ) and 1682 and 1512 cm (CONH); the NMR spectrum (CDCl ₃ ) showed that this material was essentially the title compound with signals at τ 4.20 (C ₇ -H, double doublet, J 5 and 9 Hz), 5.06 (Cg-H, doublet , J 5 Hz), 6.4 to 7.3 (C ₃ -H and C ₃ -CH ₂ CH-, unresolved 5-proton envelope), 7.84 and 7.96 (COCH _-3 , keto- and enol tautomers) and 8.59 [(CH ₃ J ₃ C-], together with diphenylmethyl-3- (2 ^ -tert.-butoxycarbonyl-3-oxobutyl) -7ß- (2-thienylacetamido) - ceph-2-em-4-carboxylate (approx. 30 %) with signals at 4.20 (C ₇ -H, double doublet, -J 5 and 9 Hz), 4.90 (Cg-H, doublet, J 5 Hz), 5.24. (C ₄ -H), 7.88 (-COCH ₃ ) and 8.55. [- (CH ₃ ) ₃ C-]; [C ₃₅ H ₃₆ N ₂ O ₆ S ₂ -H ₂ O (658.3) ϊ.

109843/19 5 4

21U541

calculated: C 63.9 H 5.7 N 4.3 S 9.7% found: 63.8 5.7 3.9 9.95%].

Example 20

Diphenyl methyl-3- (2-benzoylethyl) -7ß-aminoceph-3-e m -4- carboxylate, p ~ toluenesulfonic acid salt 2

A stirred solution of 363 mg (0.583 mmol) of diphenylmethyl-3- (2-benzoylethyl) -7β- (2-thienylacetamido) -ceph-S-em-l-carb- • ox. at and 0.56 ml (6.996 mmol) of pyridine in 10 ml of dry methylene chloride was cooled to -15 ° C and treated over the course of minutes with a suspension of 364 mg of phosphorus pentachloride (1.749 mmol) in 10 ml of dry methylene chloride, the temperature rising to -10 to -12 ° was maintained. The solution was then stirred at -10 ° C. for 30 minutes, cooled to -20 ° C. and then 5 ml of methanol were added at such a rate that the temperature did not rise above -12 ° C., and the temperature was then left to 23 ° C, the solution was stirred for 4 hours, 20 ml of 1N hydrochloric acid were added and the two-phase system was stirred for 10 minutes. The organic phase was then separated off, stirred vigorously with 30 ml of saturated sodium bicarbonate solution for 30 minutes, washed with 30 ml of water and 30 ml of brine, dried over magnesium sulfate and evaporated to 455 mg of a resin. This material was purified by chromatography over 0.05-0.2 mm silica gel (Merck) (25 g) with benzene / ethyl acetate (5/1) and then benzene / ethyl acetate (2/1) as the eluent. The combination of those fractions with an R, .- value of 0.35 (thin layer chromatography, silica gel GFP. ₅ plates, benzene / ethyl acetate (2 / l) as eluant) gave an oily solid (144.mg). A solution of this solid in 1 ml of ethyl acetate and 2 ml of ether was treated with 114 mg (0.6 mmol) of p-toluenesulfonic acid in 1 ml of ethyl acetate, then 2 ml of further ether were slowly added and the solution was cooled so that white crystals of the Salt (127 mg, 32.5%) ,. F · - 128 to 132 ° C (dec.), [A] ^ ⁵ -2.3 ° (c 0.95, CHCl ₃ ), UV spectrum \ _max (EtOH) 244 nm (B 18000), IR- spectrum

109843/1954

21H541

(CHBr ₃ ) 2640 (NH ₃ ⁺ ), 1788 (ß-lactam), 1720 (CO ₃ R) and 1684 era " ¹ (COPh), NMR spectrum " i (CDCl ₃ ) 2.18 and 2.97 ( P-CH ₃ C ₅ H ₄ SO ₃ "), 3.10 (Ph ₂ CH), 5.03 (C ₇ -H, double doublet), 5, O8 (Cg-H doublet), 6.50 and 7 , 25 (C ₂ -H, AB-quartet, 'J 16 Hz), 7.10 (4-proton envelope, -CH ₂ CH ₂ COPh) and 7.82 (P-CH ₃ -C ₆ H ₄ -) ; C ₃₆ H ₃₄ N ₂ O ₇ S ₂ -V ₂ H ₂ O:

calculated: C 63.6 H 5.2 N 4.1 S 9.4 % found: 63.6 5.2 3.6 9.5 %]

The biological results obtained are summarized in the following table.

109843/1954

Tube dilution test (v / ml) G η .tiv CvI in Gram-negative ο τ- » Protected 9 VD Grief-posj ID (D ■ Η O ω m (ED ₅₀ / mg / kg / <6 O VD rH CO CO si dose <6 CO U) ε <6 (M w ιη CNJ <6 Φ in ε • Η (Q 50 U • rl - 3 η (D ,-1 (ti υ ε • Η W. S. <u • Η U JC ε * «J (U Φ it •H (TJ • ^ t • Η J3 r-i ■ Η 4Y α • Η -P E. (Ο O Φ Ui φ υ • Η 0.6 <0.5 O « · ' • α 0.16 0.08 <0.5 -μ υ x: • W. 2.5 0.16 2 W. 125 CU α ω • Η 0.6 1.25 <0.5 16 8th 31 m approx. 50 φ 0.8 0.6 0.5 4th ω 25Ο 16 4Y approx. 50 CQ 0.8 <0.05 2 2 125 62 2 250 cn -50 Ib 1.25 0.2 1 4th 62 <4 4th 16 approx. 50 - 2 B 2.5 0.16 8th 8th 2 > 25Ο 62 31 approx. 50 - 3 2.5 0.16 1 8th 8th ) 25Ο 62 . 12th > 100 4th 0.6 0.6 2 ω, 5 16 125 > 25Ο > 50 - 5 1.25 0.08 4th 2 62 > 25Ο 125 - - 6th 1.25 0.6 2 4th 25Ο 250 > 25Ο - 7th 0.4 4, 250 > 25Ο 250 - 8th 4th > 25Ο > 25Ο 125 - 9 <0.5 125 > 25Ο - 12b 4th > 25Ο 15b 4th 250 16b

109843/1954

Pharmaceutical examples

A. Solution for intramuscular injection Composition of a single injection sample

• 7ß- (D-2-amino-2-phenylacetamido) -3-

(3-oxobutyl) -ceph-3-em-4-carboxylic acid 250 mg

Sodium chloride. 0.8 w / v%

Water for injection to 2 ml.

The solids were dissolved in water and the resulting solution filtered through a No. 5/3 sintered glass filter sterilized. The sterilized solution was made in 2.2 ml portions "Distributed in 2 ml ampoules and the ampoules hermetically sealed. The vials were then subjected to sterility testing in the usual manner.

B. Tablet -

a) 7β- (D-2-Amino-2-phenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid 250 mg

b) mannitol 75 mg

c) potato starch. 46 mg

d) corn starch 25 mg

e) Magnesium stearate 4 mg

k The dry ingredients a, b and c were mixed and granulated with a 10% aqueous paste from d. The granules were sieved through 1.405 mm (# 12 mesh BS) sieves, dried to constant weight, and sieved through a 1.003 mm (16 mesh BS) sieve. The granules were then lubricated by mixing in e and compressed into tablets weighing 400 mg with suitable presses. If necessary, the tablets can, for example, be coated with a readily soluble, customary film coating.

109843/1954

ORIGINAL INSPECTED

C. Capsule

7ß_ (D-2-Artiino-2-phenylacetamido) -3- · (3-oxobutyl) -ceph-3-em-4-carboxylic acid 250 mg

Aerosil Compositum 3 mg

The dry powders were mixed homogeneously and filled into well-fitting hard gelatin capsules so that each capsule Contained 250 mg of the active ingredient.

A silica / starch blend available from Bush, Beach & Gent, Marlon House, Lloyd's Avenue, London, E.C. 3.

109843/1954

Claims (73)

- 74. Claims 1 2 3 wherein R is a carboxylic acyl group, R and R, the Can be identical or different, hydrogen, the lower alkyl, phenyl, substituted phenyl, lower Alkoxycarbonyl-, mono- or diary1-lower-alkoxycarbonyl-, lower alkyl, carbonyl, aryl-lower alkyl 4 or C1 or C1 cycloalkyl group and R denotes hydrogen, the lower alkyl, phenyl, substituted phenyl, aryl, lower alkyl or C ₅ or C6 cycloalkyl group, as well as their non-toxic derivatives . 2.) Compounds according to claim 1, characterized in that 2 3 4 that when one of the groups R, R and R is the lower alkyl group means that the lower alkyl group is the methyl or ethyl group. 3.) Compounds according to claim 1, characterized in that 2 3 4 that when one of the groups R, R and R is a substituted phenyl group, the substituent (s) one or more halogen atoms, lower alkyl groups, lower Alkoxy groups, nitro groups, amino groups or lower alky! Amino groups. 4.) Compounds of the general formula R ^- -CHCONH-I Γ Ί · "5-C-CO-R ^ 1Q9843W54 ^r3 21U541 - where R is an aromatic group or a heterocyclic group, X is the amino, substituted amino, hydroxy, Formyloxy, lower alkanoyloxy group or water 2 3 4 Substance and R, R and R have the meanings given in claim 1, as well as their non-toxic derivatives. 5.) Vorbindungen according to claim 4, characterized in that R is the phenyl group, the phenyl group substituted with halogen atoms, hydroxy, low alkyl, nitro, amino, low alkanoyl, low alkoxy or low alkylmercapto groups, a 5 - Or 6-membered heterocyclic group which contains at least one heteroatom, such as S ₉ N or 0, or denotes a naphthyl group. the group R denotes the thien-2-yl or thien-3-yl group 6.) Compounds according to claim 5, characterized in that the
tet. 7.) Compounds of the general formula CH-CONH-T T "^, R ² J- ^^ κ> CH ₉ C-COR (III) ^} T p3 2 3 4 wherein R, R and R have the meanings given in claim 1 and their non-toxic derivatives. 8.) 7β- (D-2-Amino-2-phenylacetamido) -3- {2f-ynethyl-3-oxopentyl) -ceph-3-em-4-carboxylic acid CH ₂ -CH-CO-CH ₂ CH _{3 (IV} ) 1 09843/1954 9.) Compounds of the general formula R ⁵ -CHCONH 5 1 wherein R and X have the meanings given in claim 4, as well as their non-toxic derivatives 10.) 7β- (D-2-amino-2-phenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid CH-CONH- CO, 11.) Compounds according to any one of the preceding claims, characterized in that the non-toxic derivatives Base salts (if available) or acid addition salts (if available) are. 12.) Sodium 3- (2 | -methyl-3-oxopentyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate. 13.) 7β- (D-2-Amino-2-phenylacetamido) -3- (2 £ - methyl-3-oxopentyl) -ceph-3-em-4-carboxylic acid trifluoroacetate. 14.) 3- (2 ^ -ethoxycarbonyl-3-oxobutyl) -7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid. 15 ..) 7β- (D-2-Amino-2-phenylacetamido) -3- (3-oxobutyl) -ceph-3-em-4-carboxylic acid trifluoroacetate. 109843 / 195A ORIGINAL iKSFEGTED 16.) 3- (3-Oxobutyl) ~ 7β- (2 ~ thienylacetamido) -ceph-3-em-4-carboxylic acid. 17.) 3- (2-Acetyl-3-oxobutyl) ~ 7β- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid. 18.) 3 ~ (2 ~ Benzoylethyl) ~ 7ß- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid. 19.) 3 ~ (2t-Formylpropyl) ~ 7ß- (2 ~ thienylacetarnido) -ceph-3-em-4-carboxylic acid. 20.) 3- (2-Formylisobutyl) -7ß- (2-thienylacetamido) -ceph-3-om- 4-carboxylic acid. . 21.) 3- (2 ^ -tert _# -Butoxycarbonyl) -2-oxobutyl) -7β- (2-thienylacet amido) -ceph-3-em-4-carboxylic acid. 22.) 3- (2v-Diphenylmethoxycarbonyl-3-oxobutyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylic acid. 23.) 3- (2 ^ -Benzoyl-2 ^ -ethoxycarbonyläthvl) -7ß- (2-thienylacet ~ amido) -ceph-3-em-4-carboxylic acid. 24.) Compounds of the formula wherein R, R and R have the meaning given in claim 1 f \ * 7 and R and R, which may be identical or different, are low (C ₁ -C ₄ ) -alkyl groups, and their salts. 109843/1954 -78- 21U541 25.) Compounds according to claim 24, characterized in that fi 7 that the groups R and R represent the methyl or ethyl group mean. 26.) Pharmaceutical composition, characterized in that that it contains a compound according to one of the preceding claims and in a for use in human or veterinary medicine appropriate form. 27.) Pharmaceutical composition according to claim 26, characterized in that it contains a compound according to a of claims 1 to 25 together with a pharmaceutical carrier or binder. 28.) Pharmaceutical composition according to claim 26 or 27, characterized in that it is as a solution or suspension is in sterile pyrogen-free water » 29.) Pharmaceutical composition according to one of claims 26 or 27, which is suitable for topical use, thereby characterized in that it contains a compound according to any one of claims 1 to 25 together with a topical base. 30.) Pharmaceutical composition according to claim 26 or 27, which is suitable for use in veterinary medicine, characterized in that it provides a connection according to one of claims 1 to 25 together with a basic • position for the delayed or rapid release of active ingredients. 31.) Pharmaceutical composition according to one of the claims 26 to 30, characterized in that it contains more than 0.1% of a compound according to one of Claims 1 to 25. 109 8 A 37 195 4 _ 79 - ' 32.) Pharmaceutical composition according to claim 31, characterized in that it contains 10 to 99% of a compound according to any one of claims 1 to 25. 33.) Pharmaceutical composition according to one of claims 26 to 32, characterized in that it is in the form of Dose units is present. 34.) Pharmaceutical composition according to claim 33, characterized characterized in that each dose unit contains 50 to 500 mg of a compound according to any one of claims 1 to 25. 35.) Pharmaceutical composition according to one of claims 26 to 34, characterized in that it has a further contains therapeutic agent. 36.) Compounds of the general formula _t 2 , 4 ¹ Z-, CH ₉ -C-CO-R (VIII) COOH 2 3 4 wherein R, R and R have the meanings given in claim 1 and Z> S or ^> S - ^ O (α- or ß-) mean as well as their derivatives. 37.) Compounds according to claim 36, characterized in that the derivatives are esters, salts or salt esters. 38.) Diphenylmethyl-7β-amino-3- (2i-methyl-3-oxopentyl) -ceph-3-em-4-carboxylate-hydrogen-p-toluenesulfonate. 39.) Diphenylmethyl-7β-amino-3- (3-oxobutyl) -ceph-3-em-4-carboxylate-hydrogen-p-toluenesulfonate. 109843/1954 21U5A1 * 80 - 40.) Diphenylmethyl-3- (2-benzoylethyl) -7β-amino-ceph-3-em-4-carcoxylate-hydrogen-ρ-toluenesulfonate. 41.) Process for the preparation of a compound of the formula R "NH COOH R 1 2 3 wherein R is a carboxylic acyl group, R and R, dje can be the same or different, hydrogen, the lower alkyl, phenyl, substituted phenyl, lower alkoxycarbonyl, mono- or diaryl-lower alkoxycarbonyl, lower alkylcarbonyl-, aryl-lower-alkyl- or C ₅ - or Cg -cycloalkyl groups and R is hydrogen, the lower alkyl, phenyl, substituted phenyl, aryl-lower alkyl or C ₅ - or Cg -cycloalkyl group, or a non-toxic group Derivative thereof, characterized in that one A) a compound of the formula R ²
CH ₂ -C-CO-R ⁴ (VIII) cooR ⁸ R ³ where R is hydrogen or a carboxyl blocking group, - Z> S or> S -> 0 (α- or ß-) and R, R and 4th R have the meanings given above, with a Acylation smi ttel that of the acid R ¹ OH corresponds to, wherein R has the meaning given above, ·. acylated or 109843/1954 211A541 B) a compound of the formula (IX) COOR Ί 8 wherein R, R and Z have the meanings given above and X is chlorine, bromine or iodine, with an enamine of the formula R- R ' -Μι = C-R ' (X) 2 3 4
where RR and R have the meanings given above q
sit and the R groups either lower alkyl groups or a divalent aliphatic group which forms a heterocyclic ring together with the adjacent nitrogen atom, converts and brings the resulting reaction mixture into contact with an aqueous medium or C) a compound of the formula (IX) COOR Ί 8 wherein R, R, Z and X have the meanings given above, reacts with an anion which is replaced by a carbanion the formula , 2 R t ¹ C - CO - R " J.3 109843 / 19B4 211454t ? 3 4 * wherein R, R and R have the meanings given above, can be represented, after which, if necessary, carries out one of the following reactions D); i) a compound in which -R is a lower alkoxycar- bonyl or mono- or diaryl-lower alkoxycarbonyl group means, decarboxylated, ii) converts a / \ -isomer into the desired / \ -isomer, iii) groups protecting an amino or carboxyl group, separating and f iv) a compound in which Z> S - ^ 0, to the desired compound, wherein Z = ^ S, reduced. 42.) Process for the preparation of a compound of the formula R ² R ^X NH-1 Τ ^ l,. CK ^ -C-CO-R ⁴ <- r COOH R ³ 1 2 3 wherein R is a carboxylic acyl group, R and R, which may be the same or different, are hydrogen, lower alkyl, phenyl, substituted phenyl, lower. Alkoxycarbonyl-, mono- or diary1-lower-alkoxycarbony1-, lower alkylcarbonyl-, aryl-lower-alkyl- or C _c - or 4 denote Cg-cycloalkyl groups and R denotes hydrogen, the lower alkyl, phenyl, substituted phenyl, aryl-lower alkyl or C ₅ - or Cg-cycloalkyl group, or a non-toxic derivative thereof characterized in that one is a compound of the formula 109843/1954 21U541 (VIII) 8th
wherein R is hydrogen or a carboxyl blocking group, 2 3 Z> S or> S -4-0 (α- or ß-) and R, R and 4th
R have the meanings given above, with a Acylating agent, that of the acid R ¹ OH corresponds, in which R has the meaning given above, acylated, after which, if necessary, one of the following Carries out reactions} i) Decarboxylation of a compound in which R is a lower alkoxycarbonyl or mono- or diary1-lower alkoxycarbonyl group means, ii) Conversion of a / | -Isomers into the desired / \ - isomers, iii) Separation of groups that have an amino or carboxyl group protect, and iv) reduction of a compound in which 2 ^ is S -0 to form a compound in which Z = ^> S. 43.) The method according to claim 42, characterized in that the acylating agent is an acid halide, an acid anhydride or a mixed anhydride, an active ester or an azide or an acid together with an esterifying agent is. 44.) Method according to claims 42 or 43, characterized in that that an acid addition salt of the 7β-amino compound or the derivatives thereof are acylated. 109843/1954 21H5A1 45.) Process according to claim 44, characterized in that the acid addition salt is a hydrogen hydrocarbon sulfonate is. 46.) Method according to one of claims 42 to 45, characterized in that the acylation is carried out in an aqueous medium is carried out with an acid halide. 47.) Method according to claim 46, characterized in that the acylation is carried out in the presence of an acid-binding agent. 48.) Method according to claim 47, characterized in that the acid-binding agent is sodium bicarbonate or calcium carbonate is. 49.) The method according to claim 47, characterized in that the acid binding agent is a tertiary amine. 50.) The method according to claim 47, characterized in that the acid-binding agent is an oxirane. 51.) The method according to claim 50, characterized in that the oxirane is a low-1,2-alkylene oxide. 52.) The method according to claim 42 for producing a connection the formula SR ² R ⁵ -CH-CO-NH-τ S ^ NH CH ₂ -C-COR ⁴ Co ₂ - 2 3 4
wherein R, R and R have the meaning given in claim 1 . lines have and R has the meaning given in claim 4, characterized in that one compound 109843/1954 COPY 21H541 using the formula or an ester, a salt or a salt ester thereof with a compound of the formula R ^ -CH-CO-Cl NH ₂ -HCl acylated in the presence of an acid-binding agent. 53.) Process for the preparation of a compound of the formula R ¹ NH R * CH ₉ -C-CO-R ' COOH Ι 2 3 where R is a carboxylic acyl group, R and R, which can be identical or different, are hydrogen, lower alkyl, phenyl, substituted phenyl, lower alkoxycarbonyl, mono- or diary1-lower alkoxycarbonyl, lower alkylcarbonyl, aryl-lower alkyl or C ^ - or C -Cycloalkyl groups and R are hydrogen, the 6th
is lower alkyl, phenyl, substituted phenyl, aryl-lower alkyl or C ₅ - or Cg -cycloalkyl group, or a non-toxic derivative thereof, characterized in that a compound of the formula 109843/1954 21U541 COOR ¹ (IX) where R is hydrogen or a carboxyl blocking group, Z> S or> S - ^ - 0 (α- or ß-), R denotes those given above Has meaning and X is chlorine, bromine or iodine, with an enamine of the formula R ' (X) 2 3 4 wherein R, R and R have the meanings given above zen and the R groups either lower alkyl groups or a divalent aliphatic group, which together with the adjacent nitrogen atom is a heterocyclic ring forms, mean, converts and the resulting reaction mixture with water or an aqueous medium in contact after which, if necessary, one of the following reactions is carried out: i) Decarboxylation of a compound in which R is the lower alkoxycarbonyl or mono- or diaryl-lower alkoxycarbonyl group means, ii) conversion of a / \ -isomers into the desired / \ isomers, iii) Removal of protective groups, the amino or carboxyl groups protect, and iv) reduction of a compound in which Z is ^ S - ^ 0 to a compound in which Z means> S. 109843/1954 54.) The method according to claim 53, characterized in that the two R groups together form a divalent aliphatic Form a group with 4 carbon atoms. 55.) The method according to claim 53, characterized in that ο
the two R groups together see forming a group with 5 carbon atoms 9
the two R groups together form a divalent aliphatic 56.) The method according to any one of claims 53 to 55, characterized in that that the aqueous medium is an aqueous acidic medium which also destroys excess enamine. 57.) Process according to one of claims 53 to 56, characterized in that it is carried out at -80 to +100 ⁰ C. 58.) The method according to claim 57, characterized in that it is carried out ^{at -20 to +90 0 C.} 59.) The method according to any one of claims 53 to 58, characterized in that that 1 to 5 molar equivalents of the enamine are used. 60.) The method according to any one of claims 53 to 59, characterized in that that the process in a lower alkanecarboxylic acid nitrile, a halogenated hydrocarbon, a lower nitroalkane, a nitro-aromatic compound, a cyclic ether, an aromatic hydrocarbon or an amide. 61.) The method according to any one of claims 53 to 60, characterized in that that the enamine by reaction of an amine 9
R 2 ^NH roit a carbonyl compound of the formula R ² - CO - R ⁴ (XI) 109843 / 19S4 21H541 2 3 4 9
wherein R _f R, R and R are those given in claim 53 Have meanings, is produced. 62.) Process for the preparation of a compound of the formula CH ^ -C-CO-R (I) COOH 1 2 3 wherein R is a carboxylic acyl group, R and R, the can be identical or different, hydrogen, the lower alkyl, phenyl, substituted phenyl, lower Alkoxycarbonyl-, mono- or diaryl-lower-alkoxycarbonyl-, lower-alkylcarbonyl-, aryl-lower-alkyl- or C ₅ - or Cg -cycloalkyl groups and R is hydrogen, the lower alkyl, phenyl, substituted phenyl, aryl-lower alkyl or C ₅ - or Cg-cycloalkyl group, or a non-toxic group Derivative thereof, characterized in that a compound of the formula R ¹ NH (IX) COOR 1 8 wherein R has the meaning given above, R is hydrogen or a carboxyl blocking group, Z ^ S or ^ S - ^ 0 (oc- <or ß-) and X is chlorine, bromine or iodine, with an anion which is replaced by a carbanion the formula R ² c - CO - R "3 1098A3 / 195A 21U541 2 3 4 in which R, R and R have the meanings given above, can be represented, reacted and then if necessary, carry out one of the following reactions: i) Decarboxylation of a compound wherein R is a lower Alkoxycarbonyl or mono- or diaryl-lower alkoxycarbonyl group means, ii) conversion of a / ^ isomer into the desired A ₁ isomer, iii). Removal of protective groups, the amino or carboxyl groups protect, and iv) reduction of a compound in which Z is ^ S - $ ■ 0 to a compound in which Z '^> S means. 63.) The method according to claim 62, characterized in that the carbanion is formed by the fact that the conjugated acid , CH - CO - R ⁴ (XI) R ³ implements with a base. 64.) The method according to claim 63, characterized in that the base is an alkali metal or alkaline earth metal hydroxide, carbonate or hydrogen carbonate. 65.) The method according to claim 62, characterized in that an enolic compound with thallium-I as the carbanion source is used. 66.) A method according to any one of claims 62 to 65, characterized, in that the reaction is carried out with the carbanion at a temperature of -80 to +80 ⁰ C. 21U5A1 67.) The method according to claim 66, characterized in that the reaction is carried out with the carbanion at a temperature of -10 to +35 ⁰ C. 68.) The method according to any one of claims 62 to 6 7, characterized in that it is in an inert solvent is carried out. 69.) The method according to claim 68, characterized in that it is in a halogenated hydrocarbon, a hydrocarbon, a cyclic or acyclic ether, dimethyl sulfoxide or an amide is carried out. 70.) The method according to any one of claims 41 to 69, characterized in that that if a compound is formed in stages A, B or C, the group in the 3-position R ² ι -CH ₂ -C-COR ⁴ 3
wherein R denotes an esterified carboxyl group, a compound is obtained by subsequent hydrolysis of the esterified carboxyl group, optionally at the same time as the decarboylation, which has the group in the 3-position R ² ι -CH ₂ -CH-COR ⁴ contains. 71.) The method according to claim 70, characterized in that the decarboxylation under mild conditions by hydrolysis is carried out with an acid or base. 3/1954 21U541 72.) The method of claim 70 or 71, characterized in that the CarboxyIierung is carried out at a temperature of -40 to +100 ⁰ C. 73.) The method according to claim 72, characterized in that the decarboxy is effected. the decarboxylation at a temperature of 0 to +40 C. 109843/1964