DK143853B - METHOD OF ANALOGY FOR THE PREPARATION OF CEPHALOSPORINE COMPOUNDS OR UNTOXIC SALTS OR HYDRATES THEREOF - Google Patents

Description

(19) DENMARK

| J | (12) PUBLICATION WRITING (11) 143853 B

DIRECTORATE OF THE PATENT AND TRADEMARKET SYSTEM

(21) Application No. 381/72 (51) Intel · O 07 0 501/22 (22) Filing Date Jan 28 1972 C 07 D 501/24 (24) Race day 28 Jan 1972 (41) Aim. available July 30. 1972 (44) Presented 19 · Oct. 1981 (86) International Application No. - (86) International Incubation Day - (85) Continuation Day - (62) Stock Application No. -

(30) Priority Jan 29 19713 3531/71 »GB

(71) Applicant GLAXO LABORATORIES LIMITED, Greenf ord, GB.

(72) Inventor Michael Gregson, GB: Martin Christopher Cook, GB:

Gordon Ian Gregory, GB.

(74) Clerk of the Office of Industrial Excellence v. Svend Schønnlng.

(54) Analogous process for the preparation of cephalosporin compounds or non-toxic salts or hydrates thereof.

The present invention relates to an analogous process for the preparation of cephalosporin compounds with the i

and Y

Claim 1 shows general formula I, wherein R, R and R have the same meanings stated or non-toxic salts or hydrates thereof. It should be noted that the compounds must have a syn configuration or be present as mixtures of the syn and anti-compound q with a content of at least 75% of the syn isomer. The method according to the invention is characterized by the characterizing part of claim 1.

The compounds of the present invention are characterized in that the acylamido group of the cephalosporin or penicillin antibiotic is an α-hydroxyiminoacylamido group or α-acyloxyimino acylamido group, the compounds being syn isomers or mixtures in which the syn isomeric form dominates.

In this specification, the vision configuration is structurally stated as follows: R.C.CO.NH-VORA and the anti-configuration as follows: R.C.CO.NH-

II

N

These configurations are attributed on the basis of a work by Ahmad and Spencer, Can. J. Chem. 1961, Volume 39, page 1340.

The term non-toxic in relation to derivatives of the subject compounds means such derivatives which are physiologically acceptable at the doses in which they are administered.

Salts which can be formed, where possible, from the compounds of the present invention are (a) inorganic base salts such as alkali metal salts, e.g., sodium and potassium salts, alkaline earth metal salts such as calcium salts and organic base salts such as procain and dibenzylethylene diamine salts; and (b) acid addition salts with, for example, hydrochloric acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, toluene-p-sulfonic acid and methanesulfonic acid. The salts may also be in the form of resinates formed, for example, with polystyrene resin containing amino groups, quaternary amino groups or sulfonic acid groups, or a resin containing carboxylic groups such as a polyacrylic acid resin. The resin may, if desired, be cross-linked, i.e. it may be a copolymer of styrene and divinylbenzene containing appropriate groups. In addition, the derivatives may take the form of chelates with a heavy metal such as iron or copper.

The cephalosporin compounds mentioned herein are named from the substance cepham (J. Am. Chem.

Soc. 1962, 84, 3400). The term cephem refers to the basic cepham structure with one double bond. The penicillin compounds mentioned herein are generally named from the substance penam (J. Am. Chem. Soc. 1953, 75, 3293).

3 143853

Many cephalosporin compounds with high bactericidal activity are known. The compounds of this invention are, as mentioned, syn-isomers or predominantly syn-isomers; it has been found that these syn isomers have higher antibiotic activity than the corresponding anti isomers. Table 1 below shows the in vitro effect of a variety of anti-isomers, determined by a tube dilution method and expressed as MIC (minimum inhibitory concentration) in μ / ml. In this and the following tables, St. for Staphylococcus. E. for Escherichia, S. for Salmonella, Pr. for Proteus, Ps. for Pseudomonas and H. for Haemophilus. The names of the connections are listed under the table; the compounds are known from Danish Patent Application No. 382/72.

Table 2 is a corresponding table showing the MIC of a number of the visual compounds prepared by the process of the invention identified by reference to the example in the present description. It can be seen that the syn compounds generally have much lower MIC values and thus much higher antibacterial activity than the anti compounds. Particular attention is paid to the cases where the sight compounds corresponding to tested anti-compounds have been tested; these cases are identified in Table 2. This is supplemented by Table 3, where as test organisms some strains other than in Tables 1 and 2 have been used and where the compounds are identified by R, Ra and Rx.

The compounds of this invention are related to various known cephalosporins. Two important known related cephalosporins are cephalexin and cephaloglycine, also called 70- (D-2-amino-2-phenylacetamido) -3-methylceph-3-em-4-carboxylic acid and 70- (D-2-amino-1 2-fenylacetamido) -3-acetoxymetylceph-3-em-4-carboxyl acid. The MIC values for these two compounds are compared at the end of Table 2 and show that the compounds prepared by the process of the invention have higher antibacterial activity than the two known. It should be noted that the MIC values for cephalexin and cephaloglycine do not derive from the same test series as those for the process of the invention; and that cephalexin and cephaloglycine are compounds used for oral administration, whereas the compounds of this invention are intended to be used by injection.

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M ° ° ° 00 H. influenza ^ to on m zae 1184E i-· tn ro _ 3 8 143853 In Tables 4 and 5 a comparison of the MIC values of the compounds of the cephalexin and cephaloglycine prepared according to the examples below is made. , in these tables a code is used as follows: 5 + indicates that the compound is one tube better than cephalexin or cephaloglycine.

++ - 2 tubes better +++ - 3 tubes better ++++ - 4 tubes better 10 +++++ - ^ 5 tubes better - indicates that the compound is one tube inferior to cephalexin or cephaloglycine.

- 2 tubes inferior --- - 3 tubes inferior 15 -! · - - 4 tubes inferior ----- -> 5 tubes inferior 0 indicates that the compound has the same effect as cephalexin or cephaloglycine.

9 143853

Table 4

Comparison of the MIC values of the compounds prepared with the examples with cephalexin.

Ex. Gram Positive Gram Negative Ή

I I H I

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6 (b) 44444 14144 41 + 14 - --- - ™ 44444 4444 NT

7 (b) 44444 44444 44444 ---- ---- --- 0 4444 444 444 9 (b) 44444 11-1-1-4 44444 4 4 444 4 1 + 14 + + 10 (b) 444+ 4 +4 +++ I lH I · --- --- - - +++ + 0 11 (b) +444+ 1 41 1+ -1 1 414 --- ----- “- 44444 44444 4 + 444 12 (b) +++++ + 44 + 4 4 + 444 - --- - 0 444 + 4 44 + 4 4444+ 13 4 + 44 + 44-144 II 41 I- 0 + 0 1+ 1 +4 - +4 14 (b) 44 + 4 + 4 + 4 + 4 +4444 - 0 4 + 4 + 4 +++++ ++ 44 + 15 (b) 44444 Mill · II 1-14 - - ---- - - +++ 44 + 4 +444 16 (b) 4114 1 141 + 4 I 141-1- ---- ----- --- - ++++ 44+ 44444 17 -I 14-1 I 1 II 14 4 1 + 14 44 0 4 + 4 + 1 +1 1+ 11 + 4 + 1 III 1 21 4 11-11 4444+ 4 1-414-0 ---- ++ + ++++ 4444+ 444+ 22 4 + 44 + 4 111+ 4 4414 0 ----- 0 0 44 + 44 44+ 444+ 23 (b) 44 + 44 4 + 44 + 44+ - - + 44 44+ 44 24 +44+ 44 + 4 + 4444+ --- - - 44 - 4 25 +4444 Mill · -1444+ ----- ----- ----- --- +4444 - 44 26 (b) + 4 + 44 4 + 4 + 4 4 + 4 + 4 4 0 4+ 0 +++++ 44 + 4 +44+ 27 -i I Η I 1 + 114 44 + 44 0 0 44 + 4444+ +44 4444 28 1114 1- 441-14 +4444 0 4+ 0 4 + 44 + 4444 4444 29 (b) +++++ +++++ + 44 + 4 0 0 ++ 44444 4 + 44 + 44 30 (b) 4-1444- 4444+ 444 + 4 - 0 0 4444+ 44 444 31 44 44 1 + 14+ 4444+ - --- - - 4444+ +4 44 + 4 32 44444 +4444 44 + 44 - - + + 4444+ 4444 444 10 143853

Table 5

Comparison of the MIC values of the compounds prepared with the examples with cephaloglycine.

3ks. Gram Positive Gram Negative

I I I

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5 (b) - 0 ++ NT

6 (b) 1 11-11- I ll I I- -H4 4+ + 0 0 I 1 1 II · I l-Η I- 7 (b) II ll I- I ll 1+ Ή-Ι + 4- - —- ++ H- +44+ 44+ 9 (b) -1 + 4-4 + + 4 + 4 + + 4 + 4 + + 4 ++ 4 + 4 + 4 + 4 + 44411- + 10 ( b) 4 + 4 ++ 4 + 4 1+ 4 + 14 + 0 0 ++ +++ + 11 (b) 4 1 + 1 + -1 + 44 + 4 + 14 + 0 4+ + 4 + 4 + Hl ll- 12 (b) 4 I 1-1 + +++++ 4 + 1 4+ ++ 0 + 4+ I 1 III 4 + 4 + 13 + 4 + 1 + "1 + 44 + 4 + 44 + +++ 4+ 44+ +444+ 14 (b) 4444+ 4444+ 4 + 11 +++ ++ 4444+ 4444+ 15 (b) 44 + 4 ++ 4 + 4 + 4 + 14 + 0 + 44+ 444+ 16 (b) 4 + 44 + 444 I I- 4444+ - - + 4 + 4 + 44+ 17 44 + 4 + 4444+ 44 + 4 + 444+ 444+ 444+ 4444+ 4444+

21 4444+ 4444+ -1-144 + 44+ 44+ 444+ 444+ I I I I I

22 -ll ll I- + 4 + 4 + 4444+ 44+ 4+ 44+ 4444+ 44+ 23 (b) + 1'll 1- I Η II + 4+ 0 44 + 1- 4+ 44+ 24+ 4444+ 44+ 0 4+ 4+ —- 25 -I 14-1 + 1444+ III II- ~ --- 0 444-1 + 26 (b) 44+ 4444+ 4444+ 444+ +4+ +4+ 44 + 4 + 444+ 27 4444+ 4444+ 44+ +4+ 44+ +44+ 4444+ 44+ 28 -II ll I -144-1 + IH ll- 44+ 44+ 44+ 144-14-444+ 29 (b) II II I- 4444+ 44+ 44+ 4+ 444+ 4444+ 4444+ 30 (b) 4444+ -1111 1- 4444+ 4+ + 44+ 4444+ 4+ 31 4+ 4444+ 4444+ 0 0 4+ 4444+ 4+ 32 II l -l I 4444+ 4444+ 4+ 4+ 444+ 4444+ 444+ 11 143853 Table 6 shows a summary of the in vivo efficacy of some selected compounds of the present invention against selected compounds. bacteria in comparison to the in vivo action of cephalexin, listed in parentheses.

Table 6 EDcjg values in vivo of certain compounds, compared to the values of cephalexin (in parentheses).

Ex. ED50 mQ / kg __St. aureus 853__E. coll 851 12 (b) 4 (1) 2 (2) 13 ~ 6 (20) ~ 3 (10) 14 (b) 20 (10) 3 (4) 29 (b) __ 15 (18) __ 0.4 (1) , 3)

The most important property of the compounds of the present invention is that they have very high stability to 3-lactamases produced by a number of gram-negative bacteria, such enzymes rapidly degrading non-stable cephalosporins and thus eliminating their antibiotic action. This stability is of particular importance for cephalosporins for injection, and therefore it is compared in Table 7 below with the stability to 3-lactamases of the best known injection cephalosporin, cephaloridine, Ν- [7β - [(2-thienyl) acetamido] ] ceph-3-em-3-ylmethyl] -pyridinium-4-carboxy-acrylate. The values given in Table 4 are determined spectrophotometrically by monitoring UV absorption for the 8-lactam ring (usually in the range 260-265 nm) after incubation in the presence of the test organism, and the values are expressed relative to the value of cephaloridine arbitrarily is set to 1 against each of the test organisms listed in the table. If a cephalosporin is unstable to a given enzyme, the β-lacquer ring is hydrolyzed and the corresponding UV absorption disappears. The table shows that all the compounds prepared by the process according to the invention are substantially more stable than cephaloridine.

12 143853

Table 7

Compound β-lactamase stability according to eg enzyme preparation or litera_P. 99_E. coli TEM K. 1 1 (b) 48 15.7 9.4 3 (d) oo 11.6 4.0 4 (e) 18.8 9.4 25.3 5 (b) 15.1 33.4 7.1 6 (b) 44 22 11 '7 (b) 49 22 32.5 9 (b). oo 104 26 10 (b) 35.8 13.3 7.1 11 (b) 112 8.0 9.0 12 (b) oo 104 54 13 98 10 7 14 (b) oo 13 12 15 (b) oo 16 4 16 (b) oo 6 1.8 17 oo 16 4.0 21 oo 8 9.0 22 17 7 11.0 23 (b) oo oo 3o 24 33 63 36 25 31 16 4.0 26 (b) oo 48 27 27 22 10 5 28 33 38 20 29 (b) 65 43 36 30 (b) 76.6 60 59 31 104 21 14 32 104 37.9 19.3 i oo oo 6.63 j »9.6 3.5 k> 30> 30 3 1 00 6.8 5.1 m> 30> 30 2.41 13 143853

According to the invention, the method is used with particular advantage for the preparation of the compounds of formula B. defined in claim 2. prepared according to Examples 1b (same compound according to Examples 2b, 18 and 19), 3d (same compound in Example 8b), 15b and 16b. It can be seen from Tables 2 and 4-5 that these compounds have high antibacterial activity and from Table 7 that they have high stability against β-lacta mazes. Another group of compounds of the invention prepared with particular advantage is that defined in claim 3 of formula F. A representative of these compounds is that prepared according to Example 6b, and also its beneficial properties appear from the aforementioned tables. Finally, according to the invention, the particularly valuable compounds of the formula as claimed in claim 4 can be prepared with particular advantage. prepared in Examples 4e and 17. Their high value is also shown in the said tables, but particular attention is paid to the information on these two compounds in Tables 4 and 5; they show that their antibacterial properties are exceedingly large in comparison with the two comparison compounds. Table 2 also shows that the MIC values of these two compounds are the lowest at all.

Thus, common to the compounds of claims 2, 3 and 4 are high and broad spectrum antibiotic activity combined with good stability against β-lactamases.

Other valuable compounds which can be prepared by the process of the invention are those of the general formulas C and D

S

RU.C.CONH ___ y "S, I f N> 9

\ J_ N / Y—CH ^^ O.COR C

^ OH / 2

COOH

RU.C.CONH_ __ / N

ϋ Γ \ o = J-nxJ_ch2ococh3 d

O.CO.R6 ioOH

Wherein R is as defined in claim 2, R represents a C 2 -C 10 alkyl or alkenyl group, and represents a C 1 -C 4 alkyl group, C 1 -C 4 chloroalkyl group, C 1 -C 4 alkoxy group, phenyl. 1 group or nitrophenyl group.

For all compounds B to F, the invention also relates to the preparation of non-toxic salts or hydrates thereof.

Other important antibiotic compounds which can be prepared by the process of the invention are those of the general formula g.

R10.C.CONH-— _y '"X

^ / _N yr, —CH0R "V

^ 0Ra ^

COOH

wherein R 10 represents a phenyl group or thien-2-yl group, Ra is as defined in claim 1 and preferably is a hydrogen atom, R "represents acetoxy, crotonyloxy, isobutyryloxy, methoxy or 5-methyl-1,3,4-thiadiazole-2 Examples of important compounds of these are the following compounds in their syn-isomeric form of 3-acetoxymethyl-73- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4 - carboxylic acid, 3-acetoxymethyl-7β- [2-hydroxyimino-2- (thien-2-yl) -acetamidoj-ceph-3-em-4-carboxylic acid, 73- [2-hydroxyimino-2- (thien-2-yl) yl) -acetamido] -3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylic acid and 7 3- (2-hydroxyimino-2-phenylacetamido) -3 - (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylic acid, especially in the form of their sodium or potassium salts.

These four latter compounds are broad-spectrum antibiotics active against a wide variety of Gram-positive organisms (penicillin-resistant and penicillin-sensitive strains of Staphylococcus aureus) and Gram-negative organisms, as evidenced by in vitro and in vivo experiments. In addition, the compounds are highly resistant to β13853 β-lactamase produced by a variety of Gram-positive and Gram-negative organisms.

manufacturing

The compounds of the present invention are prepared as mentioned in the reactions (a) or (b) and any side reactions mentioned in claim 1.

In practice, it is convenient to condense an acylating agent similar to the acid of general formula VI

a (see claim 1), wherein R and R have the meanings given in claim 1, with an amino compound of formula IX also shown in claim 1, wherein Rx has the meaning given therein.

An acid addition salt of the amino compound can also be used.

Compounds where R is hydrogen can be prepared indirectly and compounds where Ra is not hydrogen can be prepared directly by using as an acylating agent an acid halide, in particular an acid chloride or acid bromide. In the preparation of a compound wherein R represents hydrogen by this technique, an additional reaction step is required, by which the acyl group R is removed to form the hydroxyimino compound. The acylation can be carried out at temperatures between -50 and + 50 ° C, preferably a temperature between -20 and + 20 ° C, e.g. 0 ° C. The acylating agent can be prepared by reacting the acid VI, wherein R is not a hydrogen atom, with a halogenating agent such as phosphorus pentachloride, thionyl chloride or oxalyl chloride. The acylation may be carried out in aqueous or non-aqueous media, and examples of suitable media are an aqueous ketone such as aqueous acetone, an ester such as ethyl acetate, an amide such as dimethylacetamide or a nitrile such as acetonitrile, or mixtures thereof.

Acylation with acid halide can be carried out in the presence of an acid-binding agent such as tertiary amine such as triethylamine or dimethylaniline, an inorganic base such as calcium carbonate or sodium bicarbonate or an oxirane which binds the hydrogen halide released by the acylation reaction. The oxirane is preferably a lower 1,2-alkylene oxide, for example ethylene oxide or propylene oxide.

16 143853

By using the free acid one obtains the compound of the general formula VI and, if desired, where R = H, are suitable condensing agents for use in the preparation of the compounds of the present invention. carbodiimides, e.g., N, N'-diethyl, dipropyl or diisopropylcarbodiimide, Ν, Ν'-di-cyclohexylcarbodiimide or N-ethyl-N'-Y-dimethylaminopropylcarbodiimide; a suitable carbonyl compound such as, for example, carbonyl-diimidazole; or an isoxazolinium salt such as N-ethyl-5-phenyl-isoxazolinium-3'-sulfonate and N-t-butyl-5-methylisoxazolinium perchlorate. The condensation reaction is conveniently carried out in an anhydrous reaction medium, for example methylene chloride, dimethylformamide or acetonitrile, since it is possible to more precisely control the reaction parameters such as temperature.

Optionally, the acylation may also be carried out with other amide-forming derivatives of the free acid, such as a symmetrical anhydride or mixed anhydride, for example with pivalic acid or formed with a halogen formate, for example a lower alkyl halide formate. The mixed symmetrical anhydrides can be formed in situ. For example, a mixed anhydride can be formed using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroguinoline. Mixed anhydrides may also be formed with phosphoric acids (e.g. phosphoric acid or phosphoric acid), sulfuric acid or aliphatic or aromatic sulfonic acids (e.g. p-toluenesulfonic acid). Other suitable acylating agents are activated esters, e.g., compounds of the formula

R.C.CO.Z

1 X

O, where Z is, for example, an azide group, oxysuccinimide group, oxybenztriazole group, pentachlorophenoxy group or p-nitrophenoxy group.

Compounds can be prepared where Ra = H by condensing an acylating agent similar to the acid VI, where R is not hydrogen but a group which can be easily removed to form the desired group -N ~ OH with the amino compound VIII. The group Ra is subsequently removed, possibly together with the removal of the group RX. Examples of such readily removable groups R a are acetyl, if desired with at least one electron withdrawing group on the α-carbon atom, e.g., trichloroacetyl, dichloroacetyl, monochloroacetyl, trifluoroacetyl, difluoroacetyl or monofluoroacetyl; formyl; benzhydryloxycarbonyl, benzyloxycarbonyl, t-butoxycarbonyl or 2,2,2-trichloroethoxycarbonyl. The removal of such groups can be carried out, for example, under mildly basic conditions. Thus, for example, acetyl groups can be removed by treatment with aqueous alkali. Halogenated acetyl groups can be removed by aqueous bicarbonate, and in addition, chloroacetyl can be removed using a nucleophile such as a thiourea. Benzhydryloxycarbonyl and t-butoxycarbonyl groups can be removed using trifluoroacetic acid with or without anisole. 2,2,2-Trichloroethoxycarbonyl can be removed by a reducing agent such as zinc / acetic acid or zinc / formic acid. It is understood that although the preparation of compounds having readily removable R groups represents an appropriate route to hydroxyimino compounds, a compounds containing such R groups are also encompassed by Formula I and may have desirable properties per se.

g

Compounds of general formula I wherein R = acyl can be recovered by side reaction (i) from corresponding compounds a of general formula I where R = H or from esters thereof (i.e., those in the 4-position have a group COOR1 where R · * - has the meaning given in claim 1) by acylation. The acylation may be carried out in any convenient manner, for example using an acid halide, symmetric or mixed anhydride, ketene, acylazide or carbodiimide α (if the 4-carboxy group is protected) corresponding to the acid R OH. Optionally, the acylation can also be carried out by means of a halogen formate, for example a chloroformate such as ethyl chloroformate, whereby you will recover carbonates, or by means of an isocyanate RuNCO, eg 2-chloroethyl isocyanate, whereby you will obtain carbamates where ORa has the formula RDNHCO. O-, where R ° represents a hydrogen atom or an optionally halogenated alkyl group. The acylation can be catalyzed, for example, by a base such as triethylamine, diethylaniline, pyridine, propylene oxide, magnesium oxide, sodium carbonate or calcium carbonate. The acylation can be carried out in an organic solvent. Examples of suitable solvents are halogenated hydrocarbons such as methylene chloride; cyclic ethers such as dioxane or tetrahydrofuran; nitriles such as acetonitrile; nitrocarbons such as nitromethane; esters such as ethyl acetate; or the acylating agent itself. The acylation can be carried out at a temperature of between -10 ° C and + 100 ° C, preferably at 0-50 ° C and particularly conveniently at 0-30 ° C. After the acylation has been performed, the group R 2 is removed if necessary.

If desired, the reaction b) specified in claim 1 can be followed. Particularly in cases where Rx is to be -CH 2 where Y is as defined in claim 1, the group Y may be introduced by methods described in the literature. Thus, starting materials of formula II wherein Y '= halogen can be prepared as described in BE Patent Nos. 719,711, 719,710, 734,532 and 734,533. Compounds of formula I wherein Y is the residue of a nucleophile as defined in claim 1 can be prepared by reacting a 3-acetoxymethylcephalosporin compound with a nucleophile corresponding to the group Y, e.g., pyridine or another tertiary amine as described in GB patent no. 912,541, 1,012,943, 1,059,562, 1,101,423, 1,206,305, 1,030,630, 1,082,943 or 1,082,962.

Compounds wherein Y is an azido group and is reacted with a dipolarophile can be prepared as described in GB Patent Nos. 1,057,883 and 1,211,694. The compound of the invention wherein Y is the residue of a nucleophile as defined in claim 1 can also be prepared by reacting a 3-halomethylcephalosporin with any of the nucleophiles shown in the foregoing references. and such a method is disclosed in BE Patent No. 719,711.

Compounds having a vinyl group or substituted vinyl group as substituent at the 3-position can be obtained by the method described in BE Patent No. 761,897.

If Y is a halogen atom (i.e., chlorine, bromine or iodine), the ceph-3-em compounds used as starting materials can be prepared by halogenating a 7β-acylamido-3-methylceph-3-em-4-carboxylic acid ester oxide followed by reduction of the Ιβ-oxide group later in the reaction sequence as described in BE patent 755,256.

The corresponding ceph-2-em compounds can be prepared as disclosed in NL Publication No. 6902013 by reacting a ceph-2-em-3 methyl compound with N-bromo-succinimide to form ceph-2-em. 3-brommetylforbindelsen.

If Rx is to be a methyl group, the compound can be prepared as described in GB Patent No. 957,569.

Cephalosporin compounds having an acyloxymethyl group as substituent at the 3-position can be prepared in any convenient manner; for example, they can be prepared from a cephalosporin with an S-C 2 OH group formed by side reaction (vi).

The group Y 'of compound II can be a chlorine atom, iodine atom bromine atom / or an acetoxy group, and the nucleophilic replacement reaction for introducing the desired substituent at the 3-position can be carried out as described in the aforementioned GB Patent No. 1,241,657.

If, on the other hand, Y is a hydroxy group (formed by the deacylation (vi)), the desired 3-acyloxymethylcephalosporin may be recovered by side reaction (vii) by acylation as described in GB-A-1,141,293; Here, there is described 3 preparation of an Δ-cephalosporin with a 3-acyloxymethyl substituent from a corresponding 3-hydroxymethyl analog consisting of aralkylating the 4-carboxy group, acylating the 3-hydroxymethyl group of the protected compound and then removing the aralkyl group.

The acylation may be carried out in any convenient manner using, for example, an acid chloride, anhydride anhydride or mixed / as acylating agent, preferably in the presence of an organic base such as pyridine and carrying out the reaction in solution in an inert anhydrous solvent such as methylene chloride. The acylation can also be carried out in aqueous acetone / sodium bicarbonate solution. The preferred acylating agent is the acid chloride.

The acylation reaction should be carried out as soon as possible since, under the acylation conditions, there may be about 2-alloyation to the Δ derivative, especially if an aroyl oxy group is introduced into the exocyclic methylene group at the 3-position.

Compounds of general formula IX can be used as esters. The free amino acid or an acid addition salt of the free amino acid or an ester thereof can also be used. Examples of useful acid addition salts, eg with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, toluene-p-sulfonic acid and methanesulfonic acid.

The ester can be formed with an alcohol, phenol, silanol, or stannanol with up to 20 carbon atoms which can be readily decomposed at a later stage of the overall reaction sequence.

Any esterifying group substituting the 4-carboxyl group of a compound of general formula IX is preferably formed with an alcohol (aliphatic or araliphatic) phenol, silanol, stannanol or acid which can be readily decomposed at a later stage of the reaction.

Suitable esters include compounds which, as an ester group, contain a group selected from the list below which cannot be considered as an exhaustive list of possible ester groups.

(i) -C00CR ° RpRq, wherein at least one of the groups R °, R 1 and R 2 is an electron donor such as, for example, p-methoxyphenyl, 2,4,6-trimethylphenyl, 9-anthryl, methoxy, acetoxy, or fur-2 yl. The remaining of the groups R R, Bp and R ^ may be hydrogen or organic substituent groups. Suitable ester groups of this type include p-methoxybenzyloxycarbonyl and 2,4,6-trimethylbenzyl oxycarbonyl.

(ii) -COOCR ° R ^ R ^, wherein at least one of the groups R °, R? and is an electron-withdrawing group such as, for example, benzoyl, p-nitrophenyl, 4-pyridyl, trichloromethyl, tribromethyl, iodomethyl, cyano-methyl, ethoxycarbonylmethyl, arylsulfonylmethyl, 2-dimethylsulfoniumethyl, o-nitrophenyl or cyano. The remaining groups of R 2, R 3 and R 2 may be hydrogen or organ substituting groups. Suitable esters of this type include: benzoylmethoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and 2,2,2-tribromoethoxycarbonyl.

(iii). -COOCbPbPbP, wherein at least two of the groups R °, R ^ and are hydrocarbon groups such as alkyl groups, eg methyl or ethyl, or aryl groups such as phenyl, while the remaining group thereof, if present, represents hydrogen. Suitable esters of this type are t-butyloxycarbonyl, t-amyloxycarbonyl, diphenylmethoxycarbonyl and triphenylmethoxycarbonyl.

(iv) -COORa wherein Ra represents adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl, tetrahydrofur-2-yl or tetrahydropyran-2-yl.

(v) Silyloxycarbonyl groups obtained by reacting a carboxyl group with a derivative of silanol. The derivative of a silanol is suitably a halogen silane or a silazane of the formula R 4 SiX; R42SiX2; R43Si.NR42; R43Si.NH.SiR43; R 4, Si.NH.COR4; R4 Si.NH.CO.NH.SiR4 ,; R4NH.CO.NR4 .SiR4.,; 3 4 4 3 4 3 3 or RC (OSiR _): NSiR where X represents a halogen atom and 3 4 the different groups R, which may be the same or different, represent hydrogen atoms or alkyl groups, e.g., methyl, ethyl, n-propyl or isopropyl ; aryl groups such as phenyl; or aralkyl such as benzyl groups. Preferred derivatives of silanols are silyl chlorides such as trimethylchlorosilane and dimethyl dichlorosilane.

The carboxyl group can be regenerated from an ester by any of the usual methods; for example, acid and base catalyzed hydrolysis is generally applicable as well as enzymatically catalyzed hydrolysis; however, aqueous mixtures can be poor solvents for these compounds and they can cause isomerizations, rearrangements, side reactions and general destruction so that special methods may be desirable.

Five suitable methods of deesterification are 1) Reaction with Lewis acids.

Suitable Lewis acids for reaction with the esters include trifluoroacetic acid, formic acid, hydrochloric acid in acetic acid, zinc bromide in benzene and aqueous solutions or suspensions of mercuric compounds. The reaction with the Lewis acid can be facilitated by the addition of a nucleophile such as anisole.

2) Reduction.

Suitable systems for performing reductions are zinc / acetic acid, zinc / formic acid, zinc / lower alcohol, zinc / pyridine, palladized charcoal and hydrogen, as well as sodium and liquid ammonia.

3) Attack of nucleophiles.

Suitable nucleophiles are those containing a nucleophilic oxygen atom or sulfur atom such as alcohols, mercaptans and water.

4) Oxidative methods, such as those involving the use of hydrogen peroxide and acetic acid.

5) Radiation.

If at the end of a given sequence of preparations, compounds where B is> S-KD are obtained, the necessary conversion can then be B> S, i.e. conversion to a sulfide is carried out, for example, by reducing the corresponding acyloxysulfonium salt or alkyloxysulfonium salt prepared in with, for example, acetyl chloride in the case of an acetoxysulfonium salt, the reduction being carried out, for example, by sodium dithionite or with iodide ions as in a solution of potassium iodide in a water miscible solvent such as acetic acid, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction can be carried out at a temperature between -20 and + 50 ° C.

Reduction of a 1-sulfinyl group may also be carried out by means of phosphorus trichloride or tribromide in such solvents as methylene chloride, dimethylformamide or tetrahydrofuran, preferably at a temperature between -20 ° C and + 50 ° C.

If the resulting compound is a ceph-2-em-4 ester, the desired ceph-3-em compound can be obtained by treating the former with a base.

The acid VI to which the acylating agent corresponds can be obtained by reacting the glyoxylic acid R.CO.COOH, where R has the meaning given above, or an ester thereof with R O.NH_, where R has the meaning given above.

The resulting acid or ester can then be separated into its syn and anti-isomers, for example, by crystallization, chromatography or distillation, followed if necessary by hydrolysis of the ester.

Acid VI, where Ra = H, can also be prepared by reacting an ester of the acid RC 1 COOH, where R has the aforementioned meaning, with an inorganic or organic nitrosating agent, for example an alkyl nitrite such as isopropyl nitrite, or nitrosyl chloride in the presence of an acid or base. It is preferred that excess nitrosating agent is used, e.g., a molar excess. Then the ester group may be removed if needed.

Vision and anti-isomers can be distinguished by suitable techniques, for example, by their ultraviolet spectra, by thin-layer chromatography or paper chromatography, or by their nuclear magnetic resonance spectra. For example, compounds of general formula I in the DMSO-dg solution extend the doublet of amide-NH at a lower field for the syn isomers than for the anti-isomers. This can be exploited by monitoring reactions.

The antibacterial compounds of the invention can be worked up for administration in any convenient manner by analogy with other antibiotics, and pharmaceutical compositions containing an antibacterial compound of the general formula I or a non-toxic derivative such as a salt thereof (as defined) can therefore manufactured for use in human or veterinary medicine. Such compositions may be reprocessed for use in the conventional manner by any necessary pharmaceutical carriers or excipients.

The present compounds may be reprocessed for injection and may be in unit dosage form in ampoules or in multi-dose containers with preservatives added. The compositions may take the form of suspensions, solutions, emulsions in oily or aqueous liquids, and may contain adjuvants such as suspending agents, stabilizers and / or dispersants. The active ingredient may also be in powder form for reconstitution with a suitable liquid such as sterile, pyrogen-free water prior to use.

The compositions may contain from 0.1% active material upwards, preferably 10-60% thereof, depending on the method of administration. Where the compositions are dosage units, each unit will preferably contain 50-500 mg of active ingredient. The dose used to treat adult humans will preferably be in the range of 10-3000 mg, e.g. 1500 mg per day, depending on the route of administration and frequency.

Some examples serve to elucidate the method of the invention.

24 T438S3 2-Hydroxyimino acetic acids and derivatives thereof

Preparation 1 2-Hydroxyimino- (thien-2-yl) -acetic acid (syn isomer)

A cooled solution of 0.834 g of hydroxylamine hydrochloride and 1.08 g of sodium bicarbonate in 20 ml of water was added to a cooled (0 ° C) solution of 2.0 g of thien-2-yl glyoxylic acid and 1.08 g of sodium bicarbonate in 20 ml of water. After two days at 20 ° C, the solution was extracted with ether, cooled and acidified with concentrated hydrochloric acid. The resulting solid was filtered off (0.564 g) and then the filtrate was thoroughly extracted with ether. The combined extracts were washed with brine, dried and concentrated. The solid residue was triturated with benzene to give 1.05 g (51%) of the oxime (syn isomer), mp 132 ° C, kmax. (ethanol) 284 nm (ε = 9500), (nujol) 2590 and 1706 (COOH) and 1655 cm -1 (C = N). (DMSO-d6) -2.5 to -Q.5 (wide multiplet; N-OH and COOH), 2.36 (multiplet; thienyl C-5H) and 2.82 (multiplet; thienyl C-3H) and C-4H).

Preparation 2 2-Dichloroacetoxyiminophenylacetic acid (syn isomer)

To a mixture of 45 ml of methylene chloride and 10 ml of dichloroacetyl chloride was added portionwise and with stirring 5 g of 2-hydroxy-iminophenylacetic acid (the syn isomer) over approx. 15 min. The reaction mixture was solid and stirred for one hour at room temperature after the addition was complete. The reaction mixture was diluted with petroleum ether (bp 40-60 ° C) and filtered and the solid was sometimes washed with petroleum ether to remove residual acid chloride. The solid was dried under vacuum to give 8.0 g (96%) of 2-dichloroacetoxyiminophenylacetic acid, m.p. 115 ° C, max. (CHBr 3) 3470 and 1750 (COOH), 1765 cm -1 (ester). T (CDCl 3) includes 2.0 -2.7 (multiplet; aromatic protons), 3.85 (singlet; -chCl 2).

Preparation 3 2-Dichloroacetoxyimino (thien-2-yl) -acetic acid (syn-isomer) 38 g of 2-Hydroxyimino- (thien-2-yl) -acetic acid (syn-isomer) was added portionwise to a solution of stirring. 70 ml of 1A38S3 dichloroacetyl chloride in 350 dry methylene chloride at 20 ° C. Stirring was continued and after approx. 20 Mon. over time, white fibrous crystals formed. The suspension was stirred for 10 min. further and then filtered. The solid was washed with methylene chloride to give 38.5 Si (61%) of the title acid, Xmax. (EtOH) 262.5 (e = 9520), 291 nm (ε = 8580).

Preparation 4 2-Dichloroacetoxyimino- (4-chlorophenyl) -acetic acid (syn isomer)

To a solution of 1.8 ml of dichloroacetyl chloride in 15 ml of methylene chloride was added 1.5 g of 2-hydroxyimino (4-chlorophenyl) acetic acid (anti-isomer) and the suspension was stirred for two hours at room temperature.

During this period, most of the material dissolved. The solid was removed, washed with petroleum ether (boiling point 40-60 ° C) and the washing liquids and filtrate were combined, kept at 5 ° C overnight, after which the resulting solid was collected and dried to give 2.0 g (85%) of the solid. the title acid with melting point 80 ° C, Kmax. (EtOH) 258 nm (ε = ΐ228θ) max. (nujol) 1774 (ester), 1750 and 1722 cm -1 (COOH).

Preparation 5 2-Dichloroacetoxyiminonaphth-1'-ylacetyl chloride (syn isomer)

To a solution of 0.375 g of 2-hydroxyiminonaphth-1-yl acetic acid (syn isomer) in 10 ml of ethyl acetate at 0 ° C was added 0.2 ml of dichloroacetyl chloride and the solution was stirred for 30 min. at 0 ° C. After the addition of 0.36 g of phosphorus pentachloride, the mixture was stirred for 90 min. at 0 ° C. The solvent was evaporated at low temperature (about 5 ° C) and toluene was evaporated from the residue at ca. 5 ° C. The residual oil was dissolved in 5 ml of ethyl acetate and used without further purification.

Preparation 6 2-Dichloroacetoxyiminophenylacetyl chloride (syn isomer)

To a suspension of 0.276 g of the syn-isomer of 2-dichloroacetoxyiminophenylacetic acid in 10 ml of methylene chloride was added 0.208 g of phosphorus pentachloride and the mixture was stirred for one hour at room temperature 26 rmts during which period the solution occurred. After removal of the solvent under reduced pressure, benzene was evaporated from the residue and the resulting oil was liberated from the solvent in vacuo to give 0.29 g (100%) of the syn isomer of 2-dichloroacetoxyiminophenylacetyl chloride, * 4 max. (liquid film) 1770 cm ”1 (COC1 and C00R).

Preparation 7 2-Dichloroacetoxyimino- (thien-2-yl) -acetyl chloride (syn isomer) 179 ml of a 0.3M solution of phosphorus pentachloride in methylene chloride was added dropwise over 15 minutes. to a winner stirring and cooled (0 ° c) suspension of 16.7 g of syn-2-hydroxyiminothien-2-yl acetic acid in 340 ml of dry methylene chloride. After 5 min. the solution was concentrated under reduced pressure and at low temperature. The red residual oil was azeotropically treated with benzene, dissolved in ethyl acetate and used without further purification.

Preparation 8 2-Hydroxyiminonaphth-11-ylacetic acid (syn and anti-isomer)

To a solution of 4.62 g of hydroxylamine hydrochloride in 40 ml of methanol was added phenolphthalein and then methanolic sodium methoxide solution (about 5% v / r, i.e. weight / volume, weight units and volume units in the same ratio as g and ml) until a pink end point was obtained. A crystal of hydroxylamine hydrochloride was added to remove the pink color and the solution was filtered to remove sodium chloride. 10.0 g of naphtha-1'-ylglyloxylic acid was added and the mixture was heated to reflux for 30 minutes. Then it was evaporated to dryness to which 50 ml of water was added and the pH was adjusted to 1 with 2N hydrochloric acid. The resulting suspension was extracted with ethyl acetate, the combined extracts washed with water, dried and evaporated to give a pale yellow solid which was recrystallized from benzene / petroleum ether (bp 60-80 ° C) to give 2.7 g (25%) 2 -hydroxyiminonaphth-1'-ylacetic acid (anti-isomer), mp 159 ° C, Xmax. (EtOH) 279 nm (ε = 594θ), (nujol) 3245 (OH), 1701, 1712 cm -1 (ΟΟΟΗ) Λ (DMSO-d6) -3.0 to 2.2 (broad signal; OH), 1.3 -2.7 (multiplet; aromatic protons).

The mother liquor from the recrystallization was evaporated to dryness, the solid (2.2 g) was treated with a slight excess of ethereal diazomethane and the yellow color was removed by the addition of the crude 2-hydroxyiminonaphth-l'-acetic acid. The ethereal solution was washed with saturated sodium bicarbonate solution, dried and evaporated to give 2.3 g of a mixture of syli and anti-methyl-2-hydroxy-iminonaphth-1'-yl acetate. 2.0 g of the mixture was dissolved in ethyl acetate and applied to eight Merck silica gel chromatographic plates (20 cm x 20 cm). The plates were developed with a 1: 2 ether / petroleum ether mixture (boiling point 40-60 ° C) and the main component having the highest R 2 value was eluted with ethyl acetate to give 0.75 g (34%) of the vis. -isomer of methyl 2-hydroxyiminonaphth-1'-yiacetate, m.p. 110-111 ° C, kmax. (EtOH) 293.5 nm (ε = 8150), - max. (CHBr 3) 3550 (OH) and 1737 cm-1 (ester) values (CDCl 3) include 1.9 to 2.85 (multiplet; aromatic protons), 6.16 (singlet; CH 2) · A solution of 1, 10 g of the ester in 15 ml of N sodium hydroxide was left for one hour and then washed with ether, acidified with 2 N hydrochloric acid and extracted with ethyl acetate. The combined extracts were washed with water, dried and evaporated to a solid which was triturated with petroleum ether (bp 40-60 ° C), filtered and dried to yield 0.89 g (86%) of the syn isomer of the title compound. acid, m.p. 111-112 ° C, \ max. (EtOH) 292 nm (ε = 7400), max. (nujol) 2630 (OH), 1673 (COOH).

Preparation 9 Ethyl thien-3-yl glyoxylate

To a solution of n-butyllithium in ether (265 ml) at a temperature of -70 ° C was added, with vigorous stirring, a solution of 41 g of 3-bromothiophene in 50 ml of ether, the addition occurring dropwise over 30 minutes during nitrogen atmosphere.

The resulting slurry was stirred at -70 ° C for five minutes and then vigorously stirred to a solution of 110 g of diethyl oxalate in 220 ml of ether, keeping the temperature below -70 ° C during addition. After the addition, the temperature of the solution was allowed to rise to room temperature and the mixture was stirred for three hours and then poured into 2N hydrochloric acid. The organic phase is separated and combined with two essential extracts of the aqueous phase. The combined extracts were dried and evaporated to give an oil which was fractionated under reduced pressure. After distillation of excess diethyl oxalate at 63 ° C / 0.1 mm HG, the title ester was collected in an amount of 12 g (26%), boiling point 110 ° C / 0.1 mm Hgr> Jmax 1720 cm ”1 (ester ), (CDCl 3) 1.43, 2.28, 2.61 (thienyl protons), 5.56 (quartet, J 8 Hz; CH 2 CH 2), 8.58 (triplet, J 8 Hz; CH 2 CH 3).

Preparation 10

Ten en-3-yl glyoxyl acid

To a mixture of 10 g of ethyl thien-3-yl glyoxylate and 50 ml of 2N sodium hydroxide was added enough methanol to produce a homogeneous solution and then allowed to stand at room temperature for one hour. The solution was poured into water, extracted with ether, acidified with 2N hydrochloric acid and extracted with ether. The combined extracts were dried and evaporated to an oil which was triturated with benzene / petroleum ether, then the resulting solid was filtered and dried to give 7.5 g (88%), mp 63 ° C, max (nujol) 3600 and 1702 cm 1 (COOH), T (DMSO-d6) values include 1.29, 2.21, 2.38 (thien-3-yl protons).

Preparation 11 2-Hydroxyimino-2- (thien-3-yl) acetic acids

To a stirred solution of 1.15 g of hydroxyl amine hydrochloride in 15 ml of methanol containing 2 drops of phenol-phthalene was added a solution of sodium methoxide in methanol until a pink color appeared. After adding one crystal hydroxylamine hydrochloride to remove the pink color, the mixture was added to a solution of 2.0 g of thien-3-yl glyoxyl acid in 10 ml of methanol and refluxed for one hour. The methanol was evaporated to a small volume and the residue was partitioned between ethyl acetate and water. The aqueous layer was acidified to pH 1 with 2N hydrochloric acid and extracted with ethyl acetate. After drying, ethyl acetate was evaporated to give a white solid consisting of isomeric 2-hydroxyimino-2- (thien-3-yl) acetic acids, max (ethanol) 254.5 nm (ε = 9700), Vmax (CHBr 3540 (OH unbound), 3200 (OH bonded), and 1750 cm -1 (COOH), T values (DMSO-d6) include 1.78 (multiplet; thienyl protons (anti-isomer)), 2.2 to 2.7 (complex multiplet; thienyl protons).

Preparation 12 2-Dichloroacetoxyimino-2- (thien-3-yl) -acetyl chloride (syn isomer) 1.03 g of 2-hydroxyimino-2- (thien-3-yl) acetic acid mixture was added portionwise to a stirred mixture. solution of 1.8 ml of dichloroacetyl chloride in 10 ml of dry methylene chloride. After 30 minutes, excess petroleum ether (boiling point 40-60 ° C) was added to the reaction mixture and the precipitated white solid was collected, washed with petroleum ether (boiling point 40-60 ° C) and dried to 1.6 g (94%) of viscosity. -isomer of 2-dichloroacet; toxyimino-2- (thien-3-yl) -acetic acid, which was suspended in dry methylene chloride and ice-cooled.

To the suspension was added dropwise a freshly prepared solution of phosphorus pentachloride (1 equivalent) in dry methylene chloride. When all the components were dissolved, solvents were removed in vacuo at low temperature and the residual oil was azeotropically treated with benzene to give the syn isomer of 2-dichloroacetoxyimino-2- (thien-3-yl) acetyl chloride as a pale yellow oil used directly.

Preparation 13 2-Hydroxyimino-2- (fur-2-yl) acetic acids

To a stirred solution of 1.94 g of hydroxylamine hydrochloride in 30 ml of methanol containing two drops of phenolphthalein was added a solution of sodium methoxide in methanol to give a pink color. After adding one crystal of hydroxylamine hydrochloride to remove the pink color, the mixture was added to a solution of 3.0 g of fur-2-ylglyoxylic acid in methanol and refluxed for one hour. The methanol was evaporated to a small volume and partitioned between ethyl acetate and water. The aqueous layer was acidified to pH 1 with 2N hydrochloric acid and extracted with ethyl acetate. After drying, the solvent was evaporated to give a yellow solid which was collected, washed with petroleum ether (bp 40-60 ° C) and dried to give 2.7 g (81%) of an isomeric mixture of 2-hydroxyimino-2- ( fur-2-yl) acetic acids, Xmax (ethanol) 271.5 nm (ε = 12400), ,max (CHBr3) 3550 (OH), 1740 and 1705 cnf1 (C00H), δ values (DMSO-d6) include 2, 62 (doublet, J 4 Hz; furyl (anti-isomer) C 3 -H), 2.14 and 3.32 (complex signals, residual furyl protons).

Preparation 14 2-Dichloroacetoxyimino-2- (fur-2-yl) -acetyl chloride (syn isomer) 1.38 g of 2-hydroxyimino-2- (fur-2-yl) -acetic acid mixture was added portionwise to an ice-cold solution of 2.8 ml of dichloroacetyl chloride in 14 ml of dry methylene chloride. After 15 minutes of stirring, a white precipitate formed which was collected, washed successively with cold dry methylene chloride and petroleum ether (bp 40-60 ° C) and dried to give 1.5 g (63%) of syn isomer one of 2- dichloroacetoxyimino-2- (fur-2-yl) -acetic acid.

To an ice-cold stirred suspension of 1.5 g of 2-dichloroacetoxy-imino-2- (fur-2-yl) -acetic acid in 70 ml of dry methylene chloride was added dropwise a freshly prepared solution of 1.0 equivalent of phosphorus pentachloride in methylene chloride. When complete dissolution occurred, the solvent was evaporated at low temperature and the residual oil was treated azeotropically with benzene to give the syn isomer of 2-dichloroacetoxyimino-2- (fur-2-yi) acetyl chloride as a pale yellow oil used directly.

Example 1 a) t-Butyl-3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

To a solution 3.28 g of t-butyl 3-acetoxymethyl-7β-amino-ceph-3-em-4-carboxylate and 2.48 g of dicyclohexylcarbodiimide in 60 ml of methylene chloride and 20 ml of dimethylformamide were added dropwise with stirring. room temperature a solution of 1.65 g of the syn isomer of 2-hydroxyiminophenylacetic acid (Ahmad and Spencer, Can J. Chem.

1961, 39, 1340) in 20 ml of dimethylformamide and the mixture was stirred at room temperature for three hours. After filtration to remove di-cyclohexylurea, the solution was evaporated and the residue was taken up in 50 ml of ether, filtered and washed successively with saturated sodium bicarbonate solution, 2N hydrochloric acid and water. The extract was dried and evaporated to give the title ester as a foam which was dried under vacuum. Yield 1.6 g, 33.8% .SI max. (CHBr +) 3570 (OH), 1786 (β-lactam), 1740 and 1230 cm -1 (0Ac), x values (CDCl3) include 2.2 to 2.7 (multiplet; aromatic protons) 7.93 (singlet) ; CHgCO), 8.46 (singlet; t-butyl ester).

b) 3-Acetoxymethyl-7β ”(2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

A solution of 1.45 g of the syn-isomer of t-butyl-3-aceto: xymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate in 25 ml of trifluoroacetic acid was left at room temperature for 10 min. The trifluoroacetic acid was evaporated under reduced pressure and the residue was dissolved in 50 ml of ether and extracted with saturated aqueous sodium bicarbonate. The combined aqueous extracts were acidified with 2N hydrochloric acid and extracted with ethyl acetate. The combined extracts were washed with water, dried and evaporated. The residue was redissolved in 5 ml of ethyl acetate and added dropwise with stirring to 500 ml of pe. troleum ether (boiling point 40-60 ° C). The precipitated solid was filtered and dried to give 0.515 g (42%) of 3-acetoxymethyl-73- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid, the syn isomer, aD = + 56 ° (c = 0.5, DMSO), max. (EtOH) 253 nm (ε = 16600), max. (nujol) 3380 (NH and OH), 1778 (β-lactam), 1720 (COOH and acetate), 1666 and 1540 cm -1 (CONH). values (DMSO-d6) include 0.32 (doublet, J 9 Hz; NH), 2.15 to 2.7 (multiplet; aromatic protons), 7.8 (singlet; CH 2 CO).

c) Sodium 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3β-4-carboxylate (syn isomer)

To a solution of 0.5 g of syn isomer of 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid ethyl 32 143 * 53 acetate solvate in 50 ml ethyl acetate was added sodium 2-ethylhexanoate (LM solution in ethyl acetate, 1 ml). After stirring for 15 min. at room temperature, the solid was collected by filtration, washed with ethyl acetate and ether and dried to yield 0.35 g (82%) of the title sodium salt, (¾ = + 98 ° (c = ii DMSO) Xmax.) pH 6.0 phosphate buffer) 253.5 nm (ε = 18400). max (nujol) 3280 (NH and OH), 1765 (β-lactam), 1740 (OCCOCH 3), 1670 and 1550 cm -1 (CONH) .% values (DMSO-d6) include 0.24 (singlet, J 9 Hz; NH), 2.28 to 2.72 (aromatic protons), 7.99 (singlet; OCOCH 2).

Example 2 a) t-Butyl 3-acetoxymethyl-78- (2-dichloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

To a solution of 3.28 g of t-butyl 3-acetoxymethyl-7β-aminoceph-3'-em-4-carboxylate in 25 ml of ethyl acetate was added a solution of 2-diloacetic toxy imino-2-phenyl 1 ac. e l lk 1 or id (syn-isomer one) in ethyl acetate (25 ml), the addition being added dropwise, stirring. The solution became hot and a precipitate formed. After two hours, the solution was filtered, washed successively with 2N hydrochloric acid, water and saturated sodium bicarbonate solution, dried and evaporated to a yellow foam which solidified on standing and triturated with petroleum ether. There was thus obtained 4.0 g (70%) of t-butyl 3-acetoxymethyl-7β- (2-dichloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate, max. (CHBr3) 3420 (NH), 1796 (β-lactam), 1730 and 1230 (acetate), 1700 and 1512 cm -1 (CONH) S values (DMSO-d6) include 2.1 to 2.5 (multiple; aromatic protons), 2.98 (singlet; CHCl 3), 7.6 (singlet; CH 3 CO), 8.5 (singlet; t-butyl ester).

b) 3-Acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

A solution of 3.0 g of t-butyl 3-acetoxymethyl-7β- (2-dichloro-acetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate in 25 ml of trifluoroacetic acid was allowed to stand at room temperature for 10 minutes. . Excess acid was removed under reduced pressure and benzene was evaporated from the residue which was taken up in ethyl acetate and extracted into aqueous sodium bicarbonate. The aqueous phase was washed with ethyl acetate, acidified and extracted with ethyl acetate. The extracts were combined, dried and concentrated to ca. 5 ml and then added dropwise to 200 ml of petroleum ether. The solid which precipitated was filtered and dried to give 1.9 g (88%) of 3-3σθΐοχγιηθΐγ1-7β ~ (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isome -re) as a white solid, = + 68 ° (c = 0.8, DMSO), tonax. (EtOH) 254 nm (ε = 17600), Si max. (nujol) 3380 (NH and OH), 1773 (β-lactam), 1720 (COOH and acetate), 1666 and 1540 cm -1 (CONH) values (DMSO-d6) include 0.32 (doublet, J 9 Hz; NH from syn isomer), 1.00 (doubled, J 9 Hz; NH from anti-isomer, constituting about $ 10 of the product), 2.15 to 2.7 (multiplet; aromatic protons), 7.8 ( singlet; CH 2 CO).

Example 3

Sodium 3-acetoxymethyl-7,8-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate (syn-isomer) a) 2-Dichloroacetoxyimino-2- (thien 2-yl) acetyl chloride (syn isomer) 27.2 g of phosphorus pentachloride was added portionwise over approx.

20 min to a stirred and cooled (ice bath) suspension of 37.0 g of the syn isomer of 2-dichloroacetoxyimino-2- (thien-2-yl) acetic acid in 370 ml of methylene chloride. The mixture was stirred at 0 ° C for an additional 30 min, during which time all the solid dissolved. After removal of the solvent under reduced pressure, benzene was evaporated from the residue at 20 ° C and the process was repeated, yielding 2-dichloroacetoxyimino-2- (thien-2-yl) acetyl chloride (the syn isomer) as an oil used directly in the next step.

b) t-Butyl 3-acetoxymethyl-78- / 2-dichloroacetoxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate (syn isomer)

A solution of 2-dichloroacetoxyimino- (thien-2-yl) acetyl chloride (syn isomer) in 300 ml of ethyl acetate was added to a stirred solution of 43 g of t-butyl-3-acetoxymethyl-7β-aminoceph-3-em -4 ~ carboxylate and 34 ml of propylene oxide in 400 ml of ethyl acetate at 20 ° C. Initially, a solid precipitated, but it gradually dissolved. The temperature was kept between 20 ° C and 30 ° C by alternately cooling and heating the solution. After four hours, the solution was washed with 2N hydrochloric acid, saturated sodium bicarbonate solution, water and brine and dried and concentrated under reduced pressure to give the title ester as an oil which was used directly in the next step.

c) 3-Acetoxymethyl-7 (3- / 2-dichloroacetoxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

A solution of t-butyl 3-acetoxymethyl-73-, 2-dichloroacetoxy-imino (thien-2-yl) -acetainido7-ceph-3-em-4-carboxylate (syn isomer) in 20 ml of anisole was treated with 100 ml of trifluoroacetic acid. After 5 min. at 20 ° C the solution was concentrated under reduced pressure at 35 ° C. Ethyl acetate was added and the solution was concentrated again, whereupon a solid separated. Ethyl acetate cif was evaporated from the residue three more times and the solid was collected and washed with ether to give 37 g (53%) of the syn isomer of 3-acetoxy-methyl-7β- / 5-dichloroacetoxyimini-2- (thienes). 2-yl) -acetamido7-ceph-3-ene-4-carboxylic acid, max. (ethanol) 263 nm (ε = 14800), * C values (DMSO-d6) include -0.19 (doublet, J 8 Hz; NH), 3.02 (singlet; CHCl2), 7.94 (singlet; OCOCH 3) Acetoxymethyl-78- / S-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid (syn isomer) 36.1 g of syn The isomer of 3-acetoxymethyl-7β-2-dichloro-acetoxyimino-2- (thien-2-yl) -acetamido-7-ceph-3-em-4-carboxylic acid was partitioned between ca. 250 ml of ethyl acetate and 200 ml of saturated sodium bicarbonate solution. The layers were separated and the ethyl acetate solution extracted twice more with sodium bicarbonate solution. The combined extracts were washed with ethyl acetate and then covered with ethyl acetate and gently acidified with concentrated hydrochloric acid. The layers were separated and the aqueous layer was extracted twice more with ethyl acetate. The combined extracts were washed with water and brine, dried and concentrated under reduced pressure to a small volume (about 60 ml) which was added dropwise to ca. 1500 ml stirred petroleum ether (boiling point 40-60 ° C). The resulting brownish solid was collected to give 27.5 g (95%) of the syn isomer of 3-acetoxymethyl-7β-2-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em. 4-carboxylic acid.

e) Sodium 3-acetoxymethyl-78- / 2-hydroxyimino- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate (syn isomer)

A solution of 12.6 g of 2-ethylhexoate in 120 ml of ethyl acetate was added to a stirred solution of 27.3 g of the syn isomer of 3-acetoxymethyl-78- / 2-hydroxyimino-2- (thien-2 yl) -acetarnido7-ceph-3-em-4-carboxylic acid in 270 ml of ethyl acetate at 20 ° C. The mixture was cooled to 0 ° C for one hour and the light brown solid was filtered off, washed with ethyl acetate and ether and dried in vacuo to 25.1 g (87.5%) of the title sodium salt, aD = + 78.5 ° (c = 1.1 in DMSO), Xmax. (pH 6 buffer) 261 nm (ε = 15200), 285 nm (ε = 10200), T (D 2 O) 2.41 (d, 5 Hz; thienyl-C-3 H or C-5 H), 2.64 (d, J 4 Hz; thienyl C-5 H or C-3 H), 2.84 (dd, J 4 and 5 Hz; thienyl C-4 H), 4.12 (d, J 5 Hz; C-7 H), 4.78 (d, J 5 Hz) ; C-6 H), 5.08 and 5.28 (2ds, branches of a quartet, J 12 Hz; C-3 CH2), 6.33 and 6.65 (2ds, branches of a quartet, J 18 Hz ; C-2 CH 2) and 7.91 (s; OCOCH 2).

Example 4 78- (2-hydroxyimino-2-phenylacetamido) -3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylic acid (syn isomer) a) 78-formamido-3- (5-methyl-l, 3,4-thiadiazole-2-yl) -tiometyl-ceph-3-em-4-carboxylic acid

A solution of 24.0 g of 78-formamido-3-acetoxymethylceph-3-em-4-carboxylic acid and 10.56 g of 5-methyl-1,3,4-thiadiazole-2-thiol in 600 ml of M pH 6.4 phosphate buffer was heated to 60 ° C for 4 1/2 hours. The solution was cooled to 20 ° C and the pH was adjusted from 6 to 5 with phosphoric acid. The solution was extracted with ethyl acetate, the pH of the aqueous layer was brought to 2 with phosphoric acid and the product extracted into ethyl acetate. The extract was washed with brine, dried and concentrated to low volume under reduced pressure, then added to stirred petroleum ether. The resulting solid was collected and dried to 9.36 g of the title compound, 0 ° = -98 ° (c = 1 in DMSO), kmax. (pH 6 buffer) 271 nm (ε = 11400), T values (DgO + NaHCO 3) include 1.74 (s; CHO), 4.30 (d, J 4.5 Hz; C-7 H), 4.88 (d, J 4.5 Hz; C-6 H) and 7.25 (s; CH 3).

b) Diphenylmethyl-7β-formamido-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate

A solution of 8.5 g of 7β-formamido-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylic acid in 300 ml of tetrahydrofuran was treated with excess of a solution of diphenyl diazomethane in 300 ml of petroleum ether, and the mixture was kept at 20 ° C for 16 hours. A few drops of acetic acid were added and the solvents removed under reduced pressure. The resulting yellow syrup was dissolved in ethyl acetate and washed with Na triumbikar bona top solution. 7.5 g of a solid precipitated which was dried in vacuo. A sample of 5.77 g of the wet solid was dissolved in methylene chloride and the solution was dried and concentrated under reduced pressure. The residue was crystallized from methanol to give 5.08 g of the title ester, m.p. 108 ° C (dec.), Max. (EtOH) 268 nm (ε = 12400), T values (CDCl3 + slightly DMSO-d6) include 1.50 (d, J 9 Hz; NH), 1.72 (s; HCO), 6.30 (singlet; C-2 CH2) and 7.31 (singlet; CH3).

c) Diphenylmethyl-76-amino-3- (5-methyl-1,3,4-thiadiazol-2-vi) -thiomethylceph-3-em-4-carboxylate hydrochloride 1.8 ml of phosphorus oxychloride was added dropwise over 2 minutes. to a stirred and cooled (0 ° C) suspension cif 4.1 g of diphenylmethyl-7B-formamido-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4 -carboxylate in dry methanol. After 30 min. the yellow solution was concentrated under reduced pressure. Ethyl acetate was added to the residue and the resulting solid was collected and washed with ether to give the title hydrochloride, max. (EtOH) 266 nm (ε = 1130θ), (nujol) include 2590 (NH 3+), 1778 (β-lactam) and 1710 cm -1 (ester). C values (DMSO-d6) include 0.50 (wide m; NHg), 2.3 to 2.7 (m; phenyl protons), 3.02 (s; CHPh 2), 4.66 + 4.76 (m; C-7H and C-6H), 5.39 + 5.69 (2ds, branches of a quartet) , J 13 Hz; C-3 CH 2 S), 6.15 (singlet; C-2 CH 2) and 7.36 (singlet; CH 3 → d) Dif enylmethyl-7B-amino-3- (5-methyl-1) 3,4-Thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate 5 g Diphenylmethyl-7β-amino-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph -3-em-4-carboxylate hydrochloride was shaken with saturated sodium bicarbonate solution and ethyl acetate. The layers were separated and the aqueous layer re-extracted with ethyl acetate. The combined ethyl acetate extracts were washed with sodium bicarbonate solution and brine, dried and concentrated under reduced pressure. The residue was crystallized from ethyl acetate / ether to give 2.75 g of the free base specified in the title, mp 152 ° C (dec.), Kmax. (EtOH) 268 nm (ε = 9000), 4 max. (CHBr 2) include 3400 and 3335 (NH 2), 1772 (β-lactam), 1720 (ester) .f values (CDCl 3) include 2.50 to 2.75 (m; phenyl protons), 3.00 (s; CHPh 2 ), 7.31 (singlet; CH3) and 8.22 (s; NH2).

e) 76- (2-Hydroxyimino-2-phenylacetamido) -3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylic acid (syn isomer)

A 1-molar solution of the syn-isomer of 2-dichloroacetoxy-imino-2-phenylacetyl chloride in 3.8 ml of ethyl acetate was added over 5 minutes. to a stirred suspension of 1.73 g of diphenylmethyl-78-amino-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate in 20 ml of ethyl acetate containing 0 , 9 ml of propylene oxide at 20 ° C. After addition was complete, the solution was allowed to stand for one hour and then diluted with ethyl acetate and washed with 2N hydrochloric acid, sodium bicarbonate solution, water and brine. The dried solution was evaporated to 2.76 g of a red foam. 2.6 g of the foam was dissolved in 5 ml of anisole and treated with 20 ml of trifluoroacetic acid. After 5 min. the solution was concentrated under reduced pressure and the residue dissolved in ethyl acetate. The solution was washed with water and then extracted with sodium bicarbonate solution. The combined aqueous extracts were washed with ethyl acetate, acidified with 2N hydrochloric acid and extracted into ethyl acetate. The combined extracts were washed with brine, dried, concentrated under reduced pressure to a small volume and added dropwise to ca. 500 ml of stirred petroleum ether to give 1.38 g of the title acid as a solid. ocD = _79 ° (c = 1.2 in DMSO), \ max. (pH 6 buffer) 256 nm (ε = 19250), m max. (nujol) includes 3650-2100 (bonded OH), 3250 (NH), 1769 (β-lactam), 1660 + 1525 cm -1 (CONH). ^ (DMSO-d6) 0.33 (d, J 8 Hz; CQNH), 2.2 to 2.7 (m; phenyl protons), 4.12 (dd, J 5 and 8 Hz; C-7 H), 4.78 (d, J 5 Hz; C-6 H), 5.43 + 5.77 (2 ds, branches of a quartet, J 13 Hz; C-3 CH 2), 6.17 + 6.41 (2 ds, branches of a quartet, J 18 Hz; C-2 CH 2 ) and 7.30 (s; CH 3).

38 143853

Examples 5-16

General Process for Preparation of 7β- (2-Hydroxy-imino-2-arylacetamido) -3- (substituted) -methylceph-3-em-4-carboxylic acids i) Preparation of the intermediate 2-dichloroacetoxyimino esters

Method A

To a solution of the t-butyl or diphenylmethyl ester of 7β-amino-3- (substituted) methylceph-3-em-4-carboxylic acid (1 equivalent) in ethyl acetate was added dropwise and with stirring at room temperature a solution of 1-1 , 15 equivalents of the 2-dilchloroacet-oxyimino-2-arylacetyl chloride (syn or anti-isomer) in ethyl acetate. The mixture was stirred for a period of 1-2 hours during which the hydrochloride of 7β-amino-3- (substituted) methylceph-3-em-4-carboxylic acid ester was precipitated and removed by filtration. The filtrate was washed successively with 2N hydrochloric acid and water and briefly with saturated sodium hydrogen carbonate solution and brine. The organic layer was dried and evaporated to the desired 2-dichloroacetoxyimino ester as a foam or an oil.

Method B

To a solution cif 1 equivalent of the t-butyl or diphenylmethyl ester of 7β-amino-3- (substituted) methylceph-3-em-4-carboxylic acid and 3-15 equivalents of propylene oxide in ethyl acetate (however, methylene chloride was used in Example 7 ) a solution of 1.05 -125 equivalents of synis omer of one of the 2-dichloroacetoxyimino-2-arylacetyl chloride in ethyl acetate was added dropwise and with stirring at room temperature. Stirring was continued until the reaction was complete (detected by thin layer chromatography; usually 40 minutes to 2 hours) at room temperature. The reaction solution was then worked up as described under Method A of the desired ester.

Method C

A solution of 1-1.1 equivalents of the syn isomer of the 2-dichloroacetoxyimino-2-arylacetyl chloride in ethyl acetate was added dropwise to a stirred suspension of 1 equivalent diphenylmethyl-7β-amino-3-acyloxy-methylceph-3- em-4-carboxylate-p-toluenesulfonic acid salt and 0-20 equivalents of propylene oxide in ethyl acetate at room temperature. After stirring for 20 min. for 2 hours, the resulting solution was worked up as described under Method A to the desired ester.

Method D

As method C except that the reaction was carried out in a mixture of acetonitrile and dimethylacetamide instead of ethyl acetate / propylene oxide; this only applies to example 12.

ii) Removal of the protecting groups on the intermediate trainers

Method E

The t-butyl or diphenylmethyl-2-dichloroacetoxyimino ester prepared by Method A or B was dissolved in trifluoroacetic acid (7-15 mg / g ester) and left at room temperature for 5-10 minutes, then evaporated under reduced pressure. The crude product was sometimes azeotropically treated with benzene to remove traces of trifluoroacetic acid. The crude product was taken up in ethyl acetate and extracted with a saturated solution of sodium bicarbonate. The aqueous extracts were washed with ethyl acetate and then acidified with 2N hydrochloric acid and extracted into ethyl acetate. The organic layer was separated, dried and evaporated to a small volume and added dropwise to a large, well-stirred volume of petroleum ether (boiling point 40-60 ° C). The resulting amorphous solid was collected and dried in vacuo to give the desired 73- (2-hydroxyimino-2-arylacetamido) -3- (substituted) methylceph-3-em-4-carboxylic acid.

Method F

The esters prepared by methods A-D were dissolved in anisole (1-15 ml / g ester) and treated with trifluoroacetic acid (4-10 ml / g ester) at room temperature. After 5-10 min. the trifluoroacetic acid was removed by evaporation under reduced pressure and the desired acid was isolated as described under Method E.

Iii) Preparation of diphenylmethyl-7g-amino-3-acyloxymethyl-ceph-3-em-4-carboxylate-toluene-p-sulfonate salts, used as starting materials in Examples 11-14 inlcl.

a) Diphenylmethyl-3-pivaloyloxymethyl-7β- (thien-2-yl) -3-acetamidoceph-3-em-4-carboxylate

A solution of 2.4 ml of pyridine in 5 ml of dry tetrahydrofuran was added to a stirred solution of 3.12 g of diphenylmethyl-3-hydroxymethyl-7β- (thien-2-yl) -acetamidoceph-3-em-4-carboxylate and 7 2 g of pivaloyl chloride in 150 ml of dry tetrahydrofuran at 20 ° C. The solution was stirred at 20 ° C for 16 hours, filtered and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed successively with sodium bicarbonate solution, 2N hydrochloric acid, sodium bicarbonate solution, water and brine. The dried solution was concentrated under reduced pressure to a solid residue which was triturated with ether to give 2.7 g (75%) of the title ester, αβ = + 5.1 (c = 1.4, CHClC), λίηίΐ. (ethanol) 235 (ε = 1410θ), 257 (ε = 8300) and 263 nm (ε = 8050) .Vmax. (CHBr3) 1787 (β-lactam), 1722 (ester), 1680 and 1510 cm -1 (COHH) values (CDCl3) include 8.81 (singlet; C (CH3) 3).

Similarly: b) Diphenylmethyl-3-benzoyloxymethyl-78- (thien-2-yl) -acetamidoceph-3-em-4-carboxylate, yield 88%, kmax. (EtOH) 223 nm (ε = 28600), kinfl. 260 (ε = 9800) and 278.5 nm (ε = 7300). (CHBr 3) 1795 (β-lactam), 1729 (ester), 1690 and 1516 cm -1 (CONH). 17 values (CDCl3) include 2.3 to 3.1 (aromatic protons), 6.18 (singlet; thienyl-CH2), 4.62 + 5.02 (2 doublets, J 14 Hz; C-3 CHgOCO).

c) Diphenylmethyl-3-crotonoyloxymethyl-78- (thien-2-yl) -acetamidoceph-3-em-4-carboxylate, yield 52%, αβ = + 7.2 ° (c = 0.8 in CHCl3) , Inflation. (ethanol) 237.5 nm (ε = 14400) and 260 nm (ε = 8000). * >> max. (CHBr3) 1783 (β-lacquer roof), 1719 (ester), 1680 and 1510 cm "1 (CONH) .ΠΓ values (CDCl13) include 6.19 (singlet; thienyl-CHg) and 8.12 (double duplicate , J 2 and 7 Hz; CH = CHCH 3).

D) Ρί £ θην ^ θΐγ1-3-ϊ3ο ^ ΐγΓγ1οχ ^ Θΐγ1 “7β '- (ΐΐβη- · 2-νΐ) - acetamidoceph-3 ~ em-4-carboxylate, yield 75%, αβ = + 4 ° ( c = 0.9 in CHCl 3), kinfl. (EtOH) 235 (e * 13400), 257 (ε = 7900) and 261.5 nm (ε = 7750) .4 max. (CHBr 3) 1767 (β-lactam), 1710 (ester), 1667, and 1500 cm -1 (CONH) S values (CDCl 3) include 6.19 (singlet; thienyl CH 2), 8.89 (doublet, J 6.5 Hz; (CH3) 2CH).

e) Diphenylmethyl-7β-amino-3-pivaloyloxymethylceph-3-emr-4-carboxylate-toluene-β-sulfonate salt

A solution of 6.2 g of phosphorus pentachloride in 80 ml of methylene chloride was added to a cooled (-10 ° C) and stirred solution of 6.0 g of diphenylmethyl-3-pivaloyloxymethyl-7β- (thien-2-yl) -acetamidoceph-3 -em-4-carboxylate and 9.6 ml of pyridine in 80 ml of methylene chloride. After 30 min. 100 ml of methanol were added at such a rate that the temperature remained at -10 ° C. The solution was then allowed to warm to 20 ° C and after three hours at 20 ° C it was cooled to 0 ° C. 140 ml of 2N hydrochloric acid was added to the vigorously stirred solution. After stirring 4 was continued for one hour, the methylene chloride layer was separated and washed successively with 2N hydrochloric acid, sodium bicarbonate solution, water and brine, then dried and concentrated under reduced pressure.

The remaining gum was dissolved in 100 ml of ethyl acetate and treated with a solution of 1.89 g of toluene-p-sulfonic acid monohydrate in 10 ml of ethyl acetate. The cooled solution deposited crystals which were collected and washed with ethyl acetate and ether to give 4.34 g (66%) of the title salt, aD = + 5.9 ° (c = 1, DMSO), Kmax. (ethanol) 261.5 nm (ε = 7700), kinfl. 267.5 (ε = 7400) and 226.5 'nm (ε = 16600) .V max. (CHBr3) 2600-2000 (NH3 +), 1800 (β-lactam) δ: 1731 cm -1 (ester). * 5 values (CDCl3) include 2.20 + 2.98 (2 doublets, J 8 Hz; toluene sulfonate aromatic protons) and 8.89 (singlet; c (oh3) 3).

Similarly: f) Diphenylmethyl-7β-amino-3-benzoyloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonate salt needles, yield 75%, ocD = + 9.5 ° (c = 0.94 in DMSO), kmax. (ethanol) 221 (ε = 32200) and 262 nm (ε = 9100) .4 max. (nujol) 2600-2800 (NH3 +), 1804 (β-lactam) and 1730 cm cm "(ester). T values (DMS0-d6) 1 3853 42 include 4.79 + 4.99 (2 doublets, branches of a quartet, J 14 Hz; C-3 CH 2) and 7.70 (singlet; CH 3).

g) Diphenylmethyl-73-amino-3-crotonoyloxymethylceph-3-em.-4-carboxylate-toluene-p-sulfonate salt yield 54%, αβ = + 7.3 ° (c = 0.89 in DMSO), Xmax. (Ethanol)

261.5 nm (ε = 8000), \ infl. 266 nm (ε = 7700). (CHBr +) 2600-2700 (NH3 +), 1790 (β-lactam), 1720 and 1221 cm -1 (ester). Values (CDCl3) include 2.20 + 2.89 (2 doublets, branches of a quartet, J 8 Hz; toluene-p-sulfonate aromatic protons) and 8.11 (double doubled, J

6.5 and 1.5 Hz; CH-CH-CH 3).

h) Diphenylmethyl-78-amino-3-isobutyryloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonate all yield 65%, Op = + 5.3 ° (c = 1.21 in DMSO), \ max. (EtOH) 262 nm (ε = 8000),>) max. (CHBr3) 1780 (β-lactam), 1713 cm-1 (ester).

values (CDCl3) include 2.21, 2.99 (2 doublets, J 8 Hz; toluene p-sulfonate) and 8.91 (doublet, J 6 Hz; (CH 3) 2 CH).

Example 5 a) t-Butyl-78- (2-dichloroacetoxyimino-2-phenylacetamido) -3-methylceph-3-em-4-carboxylate (syn isomer)

Light yellow foam, yield 75%, ^ max. (CHBr ~) 3400 (NH), 1782-i (β-lactam), 1712 (ester), 1782 and 1693 cm (CONH). <* Values (DMSO-d6) values include 2.15-2.60 (multiplet aromatic protons), 3.03 (singlet; CHCl2), 80.3 (singlet; C-3 methyl), 8.54 (singlet; t-butyl ester) prepared by Method A.

b) 78- (2-hydroxyimino-2-phenylacetamido) -3-methylceph-3-em-4-carboxylic acid (syn isomer containing about 10% of the anti-isomer) 88%, max. (nujol) 3270 (NH), 1760 (β-lactam), 2600 and 1710 (COOH), 1670 and 1530 cm -1 (CONH) X values (DMSO-d6) include 0.36 (doubled, J 9 Hz; NH from syn isomer), 1.15 (doubled, J 9 Hz; NH from anti-isomer), 2.25-2.5 (multiplet; aromatic protons), 7.92 (singlet; CH 3), prepared by Method E.

Example 6 43 143863 el) Diphenylmethyl-78- (2-dichloroacetoxyimino-2-phenylacetamido) -3-methylthiomethylceph-3-em-4-carboxylate (syn isomer) was obtained as an oil, max, (CHBr oh), 1790 (β-1actam), 1726 (ester), 1696 and 1520 cm -1 (CONH) S values (DMSO-d6) 2.1-2.8 (multiplet; aromatic protons), 3.05 (singlet) ; diphenyl methyl ester), 4.05 (quartet, J 9 Hz and 4.5 Hz; C-7 H), 4.85 (doublet, J 4.5 Hz; C-6 H), 6.45 (wide singlet; C-2 and C-3 CH2), 8.2 (singlet; SCHg) were prepared by Method A.

b) 7β- (2-hydroxyimino-2-phenylacetamido) -3-methylthiomethyl-ceph-3-em-4-carboxylic acid (syn-isomer) O 0 = + 47.5 ° (c = 1 in DMSO), Xmax. (pH 6 phosphate buffer) 255 nm (ε = 18400), max. (nujol) 3270 (NH and OH), 1752 (β-lactam), 1660 and 1518 cm -1 (CONH) S (DMSO-d6) 0.35 (doublet, J 9 Hz; NH), 2.3-2 , 7 (multiplet; aromatic protons), 4.15 (quartet, J 9 Hz and 4.5 - 'Hz; C-7 H), 4.7 (doublet, J 4.5 Hz; C-6 H), 6.35 (broad singlet; C-2. And C-3 CH 2), 7.98 (singlet; SCH 3) were prepared by Method E.

Example 7 a) Diphenylmethyl - (2-dichloroacetoxyimino-2-phenylacetamido) -3-vinylceph-3-em-4-carboxylate (syn isomer)

Like a yellow foam, max. (CHBr3) 3500 (OH), 3380 (NH), 1770 (β-lactam), 1720 (ester), 1685 and 1510 cm -1 (CONH) values · (CDCl3)! includes 2.2-2.85 (multiplet; aromatic protons), 4.0 -5.0 '' (multiplet; C-6H, C-7H and vinyl-CH2), 6.40 (broad singlet; C -2 'CH 2) was prepared by Method B.

b) 73- (2-hydroxyimino-2-phenylacetamido) -3-vinylceph-3-em-4-carboxylic acid (syn isomer)

As a white solid in 45% yield = -72 ° (c = 0.85 in DMSO), Xmax. (pH 6 buffer) 285 nm (ε = 16400),

(nujol) 3270 (NH and OH), 1754 (β-lactam), 1710 (COOH), 1660 and 1520 cm-1 (CONH) / t "values (DMSO-d6) include 0.30 (doubled, J 9 Hz) ; NH), 2.3-2.6 (multiplet; aromatic protons), 3.03 (quartet, J 11 44 143853

Hz and 18 Hz; CH = CH 2), 4.37 and 4.66 (two doublets, J 11 Hz and 18 Hz; CH-CH 2), 6.08 and 6.41 (two doublets, branches of a quartet, J 18 Hz; C- 2 CH 2) was prepared by Method E.

Example 8 a) t-Butyl-3-acetoxymethyl-78- / 2- dichloroacetoxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate (syn isomer) was isolated as an oil by method A and the protecting groups were removed by method F to give b) 3-Acetoxymethyl-78- / 2-hydroxyimino-2- (thien-2-yl) -acetamido7- ceph-3 ~ em-4-carboxylic acid (syn -isomer) as an amorphous solid in an amount of 0.36 g, αβ = + 57 ° (c = 0.8 in DMSO), \ max. (ethanol) 263 nm (ε = 13900), max. (nujol) 3280 (NH), 1772 (β-lactam), 1720 (acetate) and 1662 and 1530 cm -1 (CONH). * Values (DMSO-d6) include 0.23 (doublet, J 8 Hz; NH) , 23.6 (double doubled, J 1 and 5 Hz; thienyl C-5 H), 2.82 (multiplet; thienyl C-3 H and C-4 H) and 7.93 (singlet; OCOCH 3).

Example 9 a) Diphenylmethyl-3-azidomethyl-7β- (2-dichloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer) was obtained as a solid, yield 46%, Xmax. (EtOH) 254 nm (e = 18000), max. (CHBr3) 3420 (NH), 2128 (N3), 1800 (β-lactam).

Τ 'values (CDCl₂) include 2.1-2.85 (multiplet; aromatic protons), 3.79 (singlet; CHCl₂) prepared by method B. Removal of protecting groups by method F gave b) 3-Azidomethyl-78 (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

A yield of 85%, ocD = + 70 ° (c = 0.84 in DMSO), \ max.

(pH 6 phosphate buffer) 253.5 nm (ε = 18800), max. (nujol) 3260 (NH), 2096 (n3), 1762 (β-lactam), 1650 and 1520 cm -1 (CONH) S values (DMSO-d6) include 0.30 (NH), 2.3-2, 65 (aromatic protons), 5.57 and 6.06 (two doublets, J 13 Hz; c-3 CH 2).

45 143853

Example 10 a) Diphenylmethyl-78- (2-dichloroacetoxyimino-2-phenylacetamido) -3-methoxymethylceph-3-em-4-carboxylate (syn isomer)

As a pale yellow foam, yield 2.31 g (9490, = + 24 ° (c = 0.8 in DMSO), Xmax. (EtOH) 256 nm (ε = 17800), max. (Nujol) 1790 (8- lactam), 1725 (COOR), 1698 and 1520 cm cm ”(amide) X values (DMSO-d6) include 2.40-2.75 (multiplet; aromatic protons), 3.75 (singlet; CHClCl), 6.80 (singlet; -OCH 3) was prepared by Method B and the protecting group was removed by Method F to give b) 78- (2-hydroxyimino-2-phenylacetamido) -3-methoxymethylceph-3-em-4-carboxylic acid (syn isomer)

As a solid, yield 55%. = + 77 ° (c = 1.02 in DMSO), kmax. (EtOH) 253 nm (ε = 16800), (nujol) 1785 (O-lactam), 1730 (COOH), 1678 and 1580 cm-1 (amide) .ΤΓ values (DMSO-d6) include 0.35 (doublet, J 9 Hz; NH), 2.3 to 2.6 (multiplet; aromatic protons), 4.14 (quartet, J 9 Hz and 4.5 Hz; C-7 H), 4.79 (doubled, J4.5 Hz; C-6 H), 6, 80 (singlet; OCH 3).

Example 11 a) Diphenylmethyl 78 ~ (2-dichloroacetoxyimino-2-phenylacetamido) -3-pivaloyloxymethylceph-3-em-4-carboxylate (syn isomer)

Watered as an oil by Method C and protection immediately removed by Method F to give b) 78- (2-hydroxyimino-2-phenylacetamido) -3-pivaloyloxymethylceph-3-em-4-carboxylic acid (syn isomer)

As a yellow solid, yield 74%. = + 62 ° (c = 1 in DMSO), \ max. (pH 6 phosphate buffer) 253.5 nm (e = 17200), max. (nujol) 3700 - 46 143 * 53 2200 (bonded OH), 1771 (β-lactam), 1710 (ester and carboxylic acid), 1665 and 1526 cm -1 (CONH) values (DMSO-d6) include 0.34 (duplicate , J 8 Hz; NH), 2.3-2.65 (multiplet; aromatic protons) and 8.82 (singlet; C (CH 3) 3).

Example 12 a) Diphenylmethyl-3-benzoyloxymethyl-70- (2-dichloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate was isolated as a mixture of syn and anti-isomers prepared by method D. Removal of the protection of the mixture by method F and separation of the acids by preparative layer chromatography on Merck silica gel 254F plates using chloroform / methanol / acetic acid 90: 7: 5 (volume) gave as the slowest moving component b) 3-Benzoyloxymethyl-7β- (2-hydroxyimino) -2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

Op = + 54.5 ° (c = 1 in DMSO), kmax. (pH 6 phosphate buffer) 236.5 nm (ε = 22200), Xin 250 nm (ε ^ 19600), "4 max. (Nujol) 3680-2200 (bound OH), 1770 (β-lactam), 1710 (carboxylic acid and ester), 1680 and 1515 cm" 1 (CONH), values (DMS0 -d6) includes 0.32 (doublet, J 8 Hz; NH), 2.00-2.50 (multiplet; aromatic protons) and 6.16 and 6.38 (2 doublets, branches of a quartet, J 18 Hz) C-2 CH 2) and the fastest moving component gave the corresponding anti-isomer in an amount of 77 mg, Xmax (pH 6 phosphate buffer) 233 nm (ε = 22500), kinfl 255 nm (ε = ΐ5000), >? max (nujol) 1770 (β-lactam), 1700 (carboxylic acid and ester), 1650 and 1520 cm (CONH). In values £> MS0-d6) include 1.01 (doublet, J 8 Hz; NH) , 1.94-2.50 (multiplet; aromatic protons) and 6.14 and 6.33 (2 doublets, branches of a quartet, J 18 Hz; C-2 CH 2).

Example 13 3-Crotonoyloxymethyl-78- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn-isomer) as an amorphous solid, yield 60%, and = + 54 ° (c = 1.0 in DMSo), kmax. (pH 6 phosphate buffer) 252 nm (ε = 16700), 4max. (nujol) 1774 (β-lactam), 1710 (COOH), 1670 and 1530 cm cm 1 (CONH).% values 47 U38S3 (DMSO-d6) include 0.35 (doublet, J 8 Hz; NH), 2, 3 to 2.7 (multiplet; aromatic protons), 3.03 (double quartet, J 15 and 7 Hz; CH = CH ~ CH 3), 4.09 (doublet, J 15 and 1 Hz; CB = CH-CH 3) , 8.14 (double doubled, J 7 and 1 Hz; CH = CH-CH 3) were prepared by a combination of methods C and F and the intermediate tester was used without characterization.

Example 14 a) Diphenylmethyl-73- (2-dichloroacetoxyimino-2-phenylacetamido) -3-isobutyryloxymethylceph-3-em-4-carboxylate (syn-isomer) was isolated as a rubber, = + 14 ° (c = 1 in DMSO) , Amax. (EtOH) 255 nm (ε = 12300), "3 max. (CHBr3) 3380 (NH), 1790 (β-lactam), 1695 and 1518 cm" 1 (CONH). T values (CDCl3) include 2.0- 2.9 (multiplet; NH and aromatic protons), 3.74 (singlet; CHCl 3), 8.88 (doublet, J 7 Hz; gem dimethyl) using Method C. Removal of the protection by Method F gave b) 7β- (2-hydroxyimino-2-phenylacetamido) -3-isobutyryloxy-methylceph-3-em-4-carboxylic acid (syn isomer) as a solid, yield 87%, α D = + 52 ° (cs 1.0 in DMSO), Amax. (EtOH) 253 nm (ε = 14100), (nujol) 3280 (NH), 1770 (β-lactam), 1660 and 1520 cm -1 (amide). Values (DMSO-d6) include 0.33 (doublet, J 9 Hz; NH), 2.3-2 , 6 (multiplet; aromatic protons) and 8.86 (doublet, J 7 Hz; C (CH 3) 2).

Example 15 a) t-Butyl 3-acetoxymethyl-78- / 2- (4-chlorophenyl) -2-dichloro-acetoxyimino-acetamido7-ceph-3-em-4-carboxylate (syn isomer) was isolated as a yellow foam by method B and the protection was removed by method F to give b) 3-Acetoxymethyl-78- / 2- (4-chlorophenyl) -2-hydroxyimino-acetamido7-ceph-3-em-4-carboxylic acid (syn isomer) a solid (16% overall yield on the 7β-amino ester), αβ = + 62 ° (c = 1.0 in DMSO), Amax. (pH 6 buffer) 258 nm (ε = 20900), max. (nujol) 3280 (NH), 1770 (β-lactam), 1660 and 1524 cm -1 (CONH). Two dyes (DMSO-d6) include 0.33 (doublet, J 9 Hz; NH) 143853 48 and 7, 98 (singlet; OCOCHg).

Example 16 a) t-Butyl 3-acetoxymethyl-7β- (2-dichloroacetoxyimino-2-naphth-11-ylacetamido) -ceph-3-em-4-carboxylate (syn isomer) was isolated as a solid, yield 70¾ , + 32 ° (c = 0.75, DMSO), \ max. (EtOH) 270 nm (ε = 13050),>) max. (CHBr ^) 3540 (OH), 3380 (NH), 1784 (β-lactam), 1738 cm ”1 (OCOCHg) λ values (CDClg) include 3.68 (singlet; CHClg), 7.93 (singlet; OCOCH ^), 8 ^ 46 (singlet; C (CH3) 3) by method B. Protection removal by method F gave b) 3 "Acetoxymethyl ~ 7β- (2-hydroxyimino-2-naphth-11-ylacetamido) -ceph-3 yield of ¾4-carboxylic acid (syn-isomer) 97 ° C = + 44 ° (c = 0.72 in DMSO), max (pH 6 phosphate buffer) 260 nm (ε = 11850), max (nujol) ) 1770 (β-lactam, 1714 cm -1 (OCOCHg) Τ values (DMSO-d6) include 0.22 (doublet, J 9 Hz; NH), 1.3-2.5 (multiplet; aromatic protons) , 7.94 (singlet; OCOCH 3).

Example 17 7- 8- / 5-Hydroxyimino-2- (thien-2-yl) -acetamido7-3- (5-methyl-1,3,4-thiadiazol-2-yl) -thiomethylceph-3-em-4 carboxylic acid (syn-isomer)

A solution of 0.585 g of 2-dichloroacetoxyimino-2- (thien-2-yl) acetyl chloride in 5 ml of ethyl acetate was added to a stirred suspension of 1 g of diphenylmethyl-7β-amino-3- (5-methyl-1,3,4). (thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate in 10 ml of ethyl acetate and 1 ml of propylene oxide at 20 ° C. After 30 min. all the solid was dissolved. The solution was washed with 2N hydrochloric acid, saturated sodium bicarbonate solution, water and brine, dried and concentrated to crude diphenylmethyl-7β- / 2-dichloroacetoxyimino-2- (thien-2-yl) -acetamido7-3- (5-methyl-1,3, 4-thiadiazol-2-yl) -thiomethylceph-3-em-4-carboxylate (the syn isomer) as a solvated foam in an amount of 1.567 g / kmax. (ethanol) 267.5 nm (E10 / ocm = 240), U values (CDCl3) include 3.00 (singlet, CHPh2), 3.75 (sirget; CHCl2) and 7.35 (singlet; CH3).

8 ml of trifluoroacetic acid was added to a solution of 1.387 g of the above ester in 2 ml of anisole at 20 ° C. After 5 min. the solution was concentrated under reduced pressure and the residue dissolved in ethyl acetate. The solvent was evaporated and again the residue 49 U3 & 63 dissolved in ethyl acetate. The solution was washed with water and then extracted with sodium bicarbonate top solution. The combined extracts were washed with ethyl acetate and then brought to pH 1.9 with concentrated hydrochloric acid and extracted with ethyl acetate. The extract was washed with water and brine, dried and concentrated under reduced pressure to a foam. Trituration with ether gave 0.50 g of the title compound as a tan solid, = -84 ° (c = 1 in DMSO), \ max.

(pH 6 buffer) 271.5 nm (8 = 17600), max. (nujol) include 3250 (NH), 1700 (β-lactam), 2600 + 1710 (COOH) and 1674 + 1548 cm -1 (CONH), δ values (DMSO-d6) 0.24 (doublet, J 8 Hz; NH), 2.37 and 2.82 (thienyl protons), 4.12 (double doubled, J 5 and 8 Hz; C-7 H), 4.76 (doubled, J 5 Hz; C-6 H), 5 , 40 and 5.75 (2 doublets, branches of a quartet, J 13 Hz; C-3 CH 2), 6.13 + 6.40 (2 doublets, branches of a quartet, J 18 Hz; C-2 CH 2) and 7.29 (singlet; CH 3).

Example 18 3-Acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

To a stirred solution of 2.72 g of 3-acetoxymethyl-70-amineceph-3-em-4-carboxylic acid and 1.85 g of sodium bicarbonate in 50 ml of water and 50 ml of acetone was added dropwise and with stirring at 5 ° C a solution of the syn isomer of 2-dichloroacetoxyimino-2-phenylacetyl chloride in 5 ml of acetone over a period of 5 min. The reaction mixture was stirred for one hour at room temperature and after this period the pH was 4.0. The acetone was removed under reduced pressure and the mixture extracted with ethyl acetate. The extracts were washed with 2N hydrochloric acid and saturated brine, dried and concentrated to ca. 5 ml. The solution was cooled to -30 ° C for 15 minutes and filtered and the solid washed with cold ethyl acetate and then petroleum ether (bp 40-60 ° C) and dried to give 1.2 g (23/6) of the title compound. indicated acid as an ethyl acetate solvate, αβ = + 64 °, \ max. (EtOH) 253 nm (ε = ΐ84θθ),.

^ max. (nujol) 3260 (NH), 1760 (β-lactam), 1710 (acetate), 1650 and 1550 cm -1 (amide), Two di is (DMSO-d6) include 0.33 (doublet, J 9 Hz; NH ) 2.3-2.65 (multiplet; aromatic protons), 7.96 (singlet; CH 3 CO).

50 U38S3

Example 19 3-Acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3, -en-4-carboxylic acid (syn isomer)

To a suspension of 4.8 g of 3-acetoxymethyl-7-aminoceph-3-em-4-carboxylic acid p-toluenesulfonic acid all in 25 ml of acetonitrile / dimethylacetamide 6: 1 was added dropwise and with stirring at room temperature a 3M solution of 2-dichloroacetoxyimino-2-phenylacetyl chloride (the syn isomer) in 5 ml of acetonitrile. The reaction mixture was stirred at room temperature until the solution was homogeneous and then poured into saturated aqueous sodium bicarbonate solution. The solution washed? with ethyl acetate, acidified to pH 1.0 with 2N hydrochloric acid and extracted with ethyl acetate. The combined extracts were washed with water and dried and concentrated to ca. 10 ml. The concentrate was added dropwise to petroleum ether (boiling point 40-60 ° C) with vigorous stirring. There was thus obtained a gummy solid which was stirred with 50 ml of ethyl acetate and the solution filtered and added dropwise to 300 ml of petroleum ether. The precipitated solid was filtered and dried to give 1.47 g (29%) of the title acid, = + 57 ° (c = 0.7 in DMSO) Kmax. (EtOH) 253.5 nm (ε = 15800).

Example 20 7 P- (2-Hydroxyimino-2-phenylacetamido) -3-methylceph-3-em-4-carboxylic acid (syn isomer)

A solution of 34.8 mg of hydroxylamine hydrochloride in 5 ml of dry methanol was neutralized to phenolphthalein with sodium methoxide in methanol. This solution was added to a solution of 201 mg of t-butyl-7β-phenylglyoxamido-3-methylceph-3-em-4-carboxylate in 10 ml of dry methanol and the mixture was stirred for 2 days at room temperature. The residue obtained by evaporation was dissolved in 20 ml of ethyl acetate and the solution washed with water, dried and evaporated to 220 mg of a yellow foam. Preparative chromatography on silica plates developed with 20% by volume ethyl acetate in benzene led to the isolation of 46 mg of the starting material, 22 mg of syn-isomer of t-butyl-7β- (2-hydroxyimino-2-phenylacetamido) -3-methylceph-3-em -4-carboxylate and 94 mg of a slow-running fraction that did not contain a β-lactam ring. 20 mg of the desired ester was shaken with 1 ml of trifluoroacetic acid at room temperature for 10 min. and then evaporated to 15 mg of the above-identified acid, identified with an authentic sample by thin layer chromatography and nuclear magnetic resonance spectrum: values (DMSO-d6) include 0.38 (doubled, J 8 Hz; NH in syn isomer ), 2.25-2.65 (multiplet; phenyl protons) and 7.95 (singlet; CH

The starting ester was prepared as follows: 0.37 g of phenylglyoxylyl chloride was added to a solution of 0.54 g of t-butyl 78-amino-3-methylceph-3-em-4-carboxylate in 10 ml of ethyl acetate and the solution was stirred at room temperature. for 1 1/2 hours, after which, according to thin-layer chromatographic study, no starting material remained. Reaction mixture was diluted with ethyl acetate and washed successively with saturated sodium bicarbonate solution, water and brine. Evaporation of the dried organic layer gave one; oil which slowly crystallized (0.80 g). Recrystallization from cyclohexane gave 0.49 g (62%) of t-butyl 7β-phenylglyoxamido-3-methylceph-3-em-4-carboxylate, mp 141-143 ° C = + 122 ° (c = 1 in ethanol) , Ό max. (0HBro) 1781 cm -1 (β-lactam), * 7 values (CDCl3) include 1.5-175 (ortho aromatic protons), 2.0-2.7 (aromatic protons),. 7.89 (singlet; CH 3), 8.45 (singlet; t-butyl).

Example 21 N- / 78- (2-Hydroxyimino-2-phenylacetamido) -ceph-3-em-3-yl-methyl7- (4-carbamoylpyridinium) -4-carboxylate (syn isomer)

A solution of 500 mg of the syn isomer of 3-acetoxynethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid and 500 mg of isonicotinamide in 10 ml of water was heated to 70 ° in 1 L / 2 hours. The solution was diluted with water, filtered to remove a small amount of soft insoluble material, and passed through a column of "Bio-rad" AG-1-X8 ion exchange resin (acetate form). The eluate was examined by electrophoresis at pH 1.9, which showed that there were. two components present which both migrated towards the cathode. The... ·. the fastest moving component was similar to isonicotinamide and the slower one had a mobility similar to cephaloridine and which gave the expected purple staining with iodine platinum reagent. Evaporation of the eluate under reduced pressure at 35 ° C gave a residue which was triturated with acetone and filtered. The solid was washed with acetone and dried to give 140 mg (25%) of the title compound. The material was homogeneous by electrophoresis at pH 1.9 and migrated toward the cathode. aD = -44 ° (c = 0.72 in DMSO), kmax. (EtOH) 251 nm (ε = 13000), * 0 max. (nujol) 3330, 3150 143853 52 (NH and OH), 1771 (β-lactam), 1680 and 1560 (CONH) and 1606 cm-1 (C00 -), T values (DMSO-d6) include 0.37, 1 , 50 (2 doublets, branches of a quartet, J 7 Hz; ρ-disubstituted pyridine ring), 1.15 and 1.76 (two broad singlets; C0NH2), 2.3-2.7 (multiplet, aromatic protons) , 4.26 (multiplet; C-7H and C-3 CH2), 4.8 (multiplet; C-6H and C-3 CH2), 6.40 and 6.88 (two doublets, branches of a quartet) , J 18 Hz; C-2 CH 2).

Example 22 β- / 7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-3-yl-methyl-7-pyridine luminal-4-carboxylate (syn-isomer)

A solution of 5 g of syn isomer of 3-acetoxymethyl-78- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid and 4 ml of pyridine in 100 ml of water was heated to 70 ° C for 3 hours. under an atmosphere of nitrogen. The reaction mixture was cooled and passed through a column of AG-1-X8 resin (acetate form) eluting with water. Fractions of 100 ml were collected and those which, by electrophoresis, were found to contain the desired product, were combined and lyophilized to give 0.75 g (14%) of β- / 7β- (2-hydroxy-imine phenylacetamido) -ceph-3-em-3-ylmethyl-7-pyridinium-4-carboxylate (the syn isomer), αβ = -49.5 ° (c = 0.83 in DMSO), Xmax. (pH 6 phosphate buffer) 255 nm (ε = 20500),>) max. (nujol) 1770 (β-lactam), 1660 and 1550 cm ”1 (COΝΗ), * (7 values (DMSO-d6) include 0.46, 1.36 and 1.80 (pyridine ring protons), 2.3-2, 7 (multiplied; aromatic protons).

Example 23 a) Diphenylmethyl-7β- (2-dichloroacetoxyimino-2-phenylacetamido) -3- (4,5-dimethoxycarbonyl-1,2,3-thiazol-1-yl) methylceph-3-em-4-carboxylic acid boxylate (syn isomer)

To a solution of 1.0 g of diphenylmethyl-3-azidornethyl-7β- (2-dichloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate in 40 ml of dioxane was added 2.5 g of dimethylacetylene dicarboxylate and the solution heated under reflux for 4 hours. The reaction mixture was cooled, the solvent was removed under reduced pressure and the residue was triturated with petroleum ether (bp 40-60 ° C).

The resulting solid was collected, washed well with petroleum ether and dried to give 1.2 g (98%) of the title ester, αβ = + 25 ° (c = 1.1 in DMSO), max. (EtOH) 53 143863 250.5 nm (ε = 19900), 3 max. (nujol) 3380 (NH), 1790 (β-lactam), 1690 and 1515 cm -1 (CONH), * Γ values (CDClCl) include 2.4-2.75 (multiplet; aromatic protons), 6.09, 6 , 21 (two singlets; methyl esters).

b) 3- (4,5-Dimethoxycarbonyl-1,2,3-thiazol-1-yl) methyl-73- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

To a solution of 1.0 g of the above ester in 8 ml of anisole was added 4 ml of trifluoroacetic acid. After standing for 10 min. at room temperature, the reaction mixture was evaporated to dryness and benzene was azeotropically removed from the residue, then dissolved in ethyl acetate and extracted with sodium bicarbonate solution, washed with ethyl acetate, acidified and extracted into ethyl acetate. The combined organic extracts were dried and concentrated to ca. 10 ml. The concentrate was added dropwise and with stirring to petroleum ether (boiling point 40-60 ° C) and the resulting precipitate filtered and dried to 0.5 g (75%) of the title acid, ocD = + 68 ° (c = 0.7 in DMSO), kmax.

(pH 6 phosphate buffer) 249 nm (ε * 22200), -3max. (nujol) 1790 (β-lactam), 1680 and 1540 cm-1 (amide). Values (DMSO-d6) include 0.31 (doublet, J 8 Hz; NH), 2.3-2.7 (multiplet aromatic protons), 4.21 and 4.48 (two doublets, J 15 Hz; C-3 CH 2), 6.07 and 6.12 (singlets, methyl esters).

Example 24 7β- (2-hydroxyimino-2-phenylacetamido) -3-hydroxymethylceph-3-em-4-carboxylic acid sodium salt (syn isomer)

To 5.0 g of the syn isomer of 3-acetoxymethyl-78- (2-hydroxy-imino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid was added sufficient saturated sodium bicarbonate solution to bring the pH 7.0, and during these periods dissolution occurred. To this solution was added 300 ml of water and 35 g of defatted wheat germ and the suspension was stirred vigorously while maintaining the pH between 6.7 and 6.9 with frequent additions of 2N sodium hydroxide. After adding approx. 6 ml, the pH remained constant at 6.9. The mixture was poured into 400 ml of acetone, filtered by diatomaceous earth and the filtrate was evaporated under reduced pressure to remove acetone and then re-filtered (diatomaceous earth). The filtrate was washed with ethyl acetate and the pH was adjusted to 2.8.

143853 54

The aqueous phase was extracted with ethyl acetate and the extracts were stirred with ca. 300 ml of water, then raise the pH to 7.0 with 2N sodium hydroxide. The phases were separated and the aqueous phase was washed twice with ethyl acetate, once with ether and evaporated under reduced pressure to remove residual solvents and then lyophilized. This resulted in 1.5 g (31%) of the title acid, = + 93 ° (c = 1.0 in DMSO), \ max. (pH 6 phosphate buffer) 252 nm (ε = 12300), max. (nujol) 1750 (β-lactam), 1650 and 1530 cm -1 (amide),% values (DMSO-d6) include 2.3 to 2.6 (multiplet; aromatic protons), 5.72 and 6.05 ( two doublets, branches of a quartet, J 13 Hz; CH 2 OH), 6.44 and 6.68 (two doublets, branches of a quartet, J 18 Hz; C-2 CH 2).

Example 25

Sodium 78- / 2-hydroxyimino-2- (thien-2-yl) -acetamido7-3-hydroxymethylceph-3-em-4-carboxylate (syn isomer)

A solution of 2.0 g of the syn isomer of sodium 3-acetoxy-methyl-73- / 2-hydroxyimino 2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate in 160 ml of water was heated to 37 ° C and 15 g of defatted wheat germ was added. The mixture was stirred at 37 ° C for 4 hours keeping the pH between 6.5 and 6.9 by the addition of 2N sodium hydroxide solution (2.3 ml total). The mixture was poured into 200 ml of acetone and then filtered through diatomaceous earth. The acetone was removed under reduced pressure and the aqueous solution was again filtered through silica gur, washed with ethyl acetate and then covered with ethyl acetate and brought to pH 2.7 by careful addition of concentrated hydrochloric acid. The layers were separated and the aqueous was re-extracted with ethyl acetate.

The combined extracts were washed with water and the pH was brought to 7.0 by the addition of 2N sodium hydroxide. The organic layer was further extracted with water and the combined aqueous solutions washed with ethyl acetate and ether, degassed and lyophilized to give 1.43 g (79%) of the title oxime as a brown solid, ocD = + 76 ° (c = 0.46, DMSO), \ max. (pH 6 phosphate buffer) 261.5 nm (ε = 10600), kinfl. 284 nm (ε = 6500),> 5 max. (nujol) 1760 (β-lactam), 1560 (CO "), 1650 and 1520 cm" 1 (COHH) / C values (d20) include 2.41, 2.63 and 2.83 (doubled, doubled and triplet, respectively). , J 4 Hz; thienyl protons).

Example 26 55 U3 & S3 a) 7β- (2-Acetoxyimino-2-phenylacetamido) -3-acetoxymethyl-c eph-3-em-4-carboxylic acid

To a solution of 8 ml of acetyl chloride in 8 ml of methylene chloride was added 500 mg of the syn isomer of 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid in portions over 10 min. under vigorous stirring at room temperature. A gum was formed and 8 ml of ethyl acetate was added to effect dissolution. After stirring for two hours at room temperature, the mixture was slowly added to 300 ml of petroleum ether with vigorous stirring. The precipitate was filtered off, washed thoroughly with petroleum ether, dissolved in 10 ml of chloroform, stirred with activated charcoal and filtered. The filtrate was added dropwise to 250 ml of petroleum ether with stirring. The precipitate was filtered off and dried to give 350 mg (62%) of the title acid, * 17 values (DMSO-d6) include -0.11 (doubled, J 8 Hz; NH), 2.2-2 , 6 (multiplet; aromatic protons), 7 »76 (singlet; Cf 2 .CO 2 N =), 7.94 (singlet; CH 2 CO). Signals at * T 0.05 (doublet, J 9 Hz; NH) and 7.78 (singlet; CH 2 CO 2 N 2) showed the presence of ca.

30% of the anti-isomer.

b) 7β "(2-Acetoxyimino-2-phenylacetamido) -3-acetoxymethylceph-3-em-4-carboxylic acid (syn isomer)

To a solution of 1.5 ml of pyridine in 20 ml of ethyl acetate was added 0.5 g of the syn isomer of 3-acetoxymethyl-7β- (2-hydroxy-imino-2-phenylacetamido) -ceph-3-em-4 carboxylic acid, and the stirred mixture was treated dropwise with a solution of 1.0 ml of acetyl chloride in 5 ml of ethyl acetate and the mixture was continued for 10 minutes. The mixture was poured into water, the ethyl acetate phase was separated and washed with 2N hydrochloric acid and water and extracted with saturated aqueous; sodium bicarbonate. The combined aqueous extracts were acidified and extracted with ethyl acetate and the extracts were dried and concentrated to ca.

5 ml. The concentrate was added dropwise with stirring to 250 ml of petroleum ether. The precipitated solid was collected and dried to give 0.32 g (58%) of the title acid, = + 83 ° (c = 0.8 in DMSO), kmax. (pH 6 buffer) 258 nm (e = 20800),> 1 max. (nujol) 3270 (NH), 1780 (β-lactam), 1680 and 1546 cm -1 ((),, values (DMSO-d6) include -0.10 (doublet, J 8 Hz; NH), 2.2 to 2.6 (multiplet; aromatic protons), 7.78 (singlet; ^ NOCOCH ^), 7.94 (singlet; ococHg).

56 143853

Example 27 3-Acetoxymethyl-7β- (2-chloroacetoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

To a solution of 7 ml of chloroacetyl chloride in 7 ml of ethyl acetate was added portionwise and with stirring 0.488 g of the syn isomer of 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid and the resulting solution was left at room temperature for two hours. It was then poured into 300 ml of petroleum ether (boiling point 40-60 ° C) with vigorous stirring. The resulting solid was washed several times with petroleum ether, filtered, washed again with petroleum ether and dried to give the title acid as a yellow solid in an amount of 0.453 g (79%), (¾ = + 52 ° (c = 0.87, DMSO), max (pH 6 phosphate buffer) 254 nm (e = -16700), max (CHBr 3) 3421 (NH), 1791 (β-lactam), 1699 and 1521 cm -1 (CONH) ,% values (DMSO-d6) include -0.13 (doublet, J 8 Hz; NH), 5.36 (singlet; CH2 Cl1), 7.96 (singlet; OCCOCH3).

Example 28 3-Acetoxymethyl-78- (2-ethoxycarbonyloxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

A solution of 0.5 g of the syn isomer of 3-acetoxymethyl-78- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid in 20 ml of saturated sodium bicarbonate solution was briefly shaken with a solution of 1 ml ethyl chloroformate in 10 ml of ethyl acetate. The aqueous layer was separated and shaken with another solution of 1 ml of ethyl chloroformate in 10 ml of ethyl acetate and then separated, washed twice with ethyl acetate, acidified, and also trailered with ethyl acetate. The combined aqueous extracts were dried and concentrated to ca. 5 ml. The concentrate was added dropwise with stirring to 200 ml of petroleum ether. The precipitate was filtered and dried to give 0.45 g (77%) of the title acid, = + 48 ° (c = 0.8 in DMSO), \ max. (pH 6 phosphate buffer) 258 nm (ε = 21800),>} max. (nujol) 3270 (NH), 1776 (β-lactam), 1679 and 1535 cm cm 1 (CONH), T values (DMSO-d6) include -0.13 (doublet, J 8 Hz; NH), 2.2 to 2.5 (multiplet; aromatic protons), 5.68 (quartet, J 8 Hz; OCH 2 CH 3), 7.96 (singlet; O COCH 3), 8.70 (triplet, J 8 Hz; OCH 2 CH 3).

Example 29 57 a) Diphenylmethyl-3-acetoxymethyl-7g- (2-benzoyloxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

To a solution of cif 1.0g diphenylmethyl-3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-quarboxylic acid and 1.5 ml of pyridine in 30 ml of ethyl acetate was added. 1.5 ml of benzoyl chloride and the mixture was allowed to stand for one hour at room temperature. The solution was then washed with 50 ml of 2N hydrochloric acid and saturated brine, dried (sodium sulfate) and concentrated to ca. 5 ml. The concentrate was added dropwise to 350 ml of petroleum ether (boiling point 40-60 ° C) and the precipitated solid was filtered, washed with petroleum ether and dried to give the title ester as a yellow solid in an amount of 1.0 g (8550, (¾ = + 6.4 ° (c = 0.94 in DMSO), kmax. (EtOH) 259 nm (ε = 26700), Smax. (CHBr ^) 1780 (β-lactam), 1640 and 1522 cm "" Values (DMSO-d6) include -0.27 (doublet, J 8 Hz; NH), 1.8 to 2.8 (multiplet; aromatic protons), 8.01 (singlet; OCOCH 3).

b) 3-Acetoxymethyl-7g- (2-benzoyloxyimino-2-phenylacetamido) -ceph-3-em ~ 4-carboxylic acid (syn isomer)

A solution of 0.85 g of the syn isomer of diphenylmethyl-3-acetoxymethyl-78- (2-benzoyloxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate in 10 ml of trifluoroacetic acid and 10 drops of anisole was allowed to stand for 10 minutes. mine. at room temperature and then evaporated to dryness under reduced pressure. The residue was dissolved in ethyl acetate and extracted with sodium bicarbonate. The combined extracts were acidified with 2N hydrochloric acid and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried and concentrated to ca.

10 ml; then they were added dropwise with stirring to petroleum ether.

The precipitated solid was filtered and dried to give 0.39 g (60%) of the title acid, = + 31 ° (c = 0.94 in DMSO), max. (EtOH) 260 nm (ε = 26000),> imax. (nujol) 3270 (NH), 1784 cm -1 (β-lactam), δ values (DMSO-d6) include -0.27 (doublet, J 8 Hz; NH), 1.75-2.6 ( multiplet; aromatic protons), 7.96 (singlet; ococh3).

The starting material used in (a) above was prepared as follows:

To a solution of 2.0 g of the syn isomer of 3-acetoxymethyl-78- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid in 50 ml of tetrahydrofuran maintained at 37 ° C was added portionwise a solution of diphenyl diazomethane in petroleum ether (boiling point 60-80 ° C) until the purple color persisted for one hour. After the addition of acetic acid to destroy excess diphenyl diazomethane, the solution was evaporated to dryness, taken up in ethyl acetate and then washed with sodium bicarbonate and brine, dried and evaporated. The residue was triturated with petroleum ether, and the resulting solid was filtered and dried to 1.93 g (69%) of diphenylmethyl-3-acetoxy-methyl-7β- (2-hydroxyimino-2-enylacetamido) -ceph-3-em-4 carboxylate (syn isomer), = + 24 ° (c = 1 in DMSO), kmax. (EtOH) 253 nm (ε = 16150), "Jmax. (CHBr +) 3610 (OH), 1796 cm -1 (β-lactam)." Values (DMSO-d6) include 0.27 (doublet, J9 Hz; NH), 8.03 (singlet; OCOCH

Example 30 a) Diphenylmethyl-3-acetoxymethyl-7β- [2- (4-nitrobenzoyloxy-imino) -2-phenylacetamido [ceph] 3-em-4-carboxylate (syn isomer)

To a solution 1.5 g of the syn isomer of diphenylmethyl-3-aceto-3-methyl-7β- (2-hydroxyimino-2-enylacetamido) -ceph-3-em-4-carboxylate and 0.5 nil of pyridine in 100 ml of ethyl acetate 0.42 g of 4-nitrobenzoyl chloride was added and the resulting solution was stirred at room temperature for one hour. Additional portions (0.042 g, 0.015 g and 0.20 g) of the acid chloride were added and the reaction mixture was stirred for a total of 4 hours at room temperature. Then the solution was washed with 2N hydrochloric acid and brine, dried, concentrated to ca. 5 ml and added dropwise and with stirring to petroleum ether (boiling point 40-60 ° C). This gave a solid which was filtered and dried to give 1g (58%) of the title ester, kmax. (EtOH) 263 nm (ε = 29000), (CHBrJ 3400 (nh), 1780 (β-lactam), 1395 and 1525 (NO2), 1687 and 1515 cm cm "(CONH). (Values (CDCl13) include 1.70 and 2.71 (two duplicates, J 8 Hz; 2.1 to 2.85 (multiplet; ΐτΗί 2 aromatic protons), 8.03 (singlet; OCOCH 2).

b) 3-Acetoxymethyl-7β- [2- (4-nitrobenzoyloxyimino) -2-phenylacetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

A solution of 1.0 g of the syn isomer of diphenylmethyl-3-acetoxymethyl-7β- [2 ~ (4-nitrobenzoyloxyimino) -2-phenylacetamido7-ceph-3-em-4 ~ k: arboxylate in 10 ml of trifluoroacetic acid at room temperature for 10 min. After evaporation of the trifluoroacetic acid under reduced pressure 14 * 3853 59, the residue was dissolved in ethyl acetate and extracted with saturated aqueous sodium bicarbonate. A solid precipitated during bicarbonate extraction and it was collected, washed with water and ethyl acetate and partitioned between ethyl acetate and 2N hydrochloric acid. The phases were separated and the upper layer was washed with water, dried, concentrated to ca. 10 ml and added dropwise with stirring to 200 ml of petroleum ether (boiling point 40-60 ° C). The precipitated solid was filtered and dried to give 0.150 g (20¾) of 3-acetoxymethyl-78- / 5- (4-nitrobenzoyloxyimino) -2-phenylacetamido7-ceph-3-em-4-carboxylic acid (syn isomer) ), α D = + 17 ° (c = 0.8 in DMSO), λ max. (EtOH) 266.5 nm. (£ = 33000), 4 max. (nujol) 3270 (NH), 1780 (β-lactam) 1520 and 1338 (NO2), 1675 and 1520 cm -1 (CONH), 'values (DM30-d6) include 1.58 and 1.69 (two duplicates) , J 8 Hz; β-substituted phenyl), 2.1 to 2.5 (multiplet; aromatic protons), 7.79 (singlet; OCOCH 2).

Examples 31 - 1 I III 11 ι »» ι · ιι II III - 3-Acetoxymethyl-7β- (2-methylcarbamoyloxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

To 5 ml of ethyl isocyanate was added 0.9 g of syn isomer of 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid ethyl acetate solvate and the mixture was stirred for one hour. at room temperature. Two additional portions of the isocyanate (5 ml) were added over a period of two hours. The mixture was then left for one hour and poured into 250 ml of petroleum ether (bp 40-60 ° C) and the resulting precipitate collected, washed thoroughly with petroleum ether and dried. The solid was dissolved in ethyl acetate and extracted into sodium bicarbonate solution; the bicarbonate solution was then washed with ethyl acetate, acidified with 2N hydrochloric acid and extracted with ethyl acetate. The combined extracts were dried and concentrated and the concentrate (about 5 ml) added dropwise to petroleum ether with vigorous stirring. The precipitate formed was filtered and dried to give 0.4 g (47¾) of the title acid, aD = + 50 ° (c = 0.85 in DMSO), Xmax. (pH 6 buffer) 257.5 nm (e = 212 ° C), -4max. (nujol) 1782 (β-lactam), 1675 and 1520 cm -1 (CONH), T values (DMSO-d6) include 0.04 (doublet, J 8 Hz; NH), 2.0 to 2.6 ( multiplet; aromatic protons), 6.7 to 7.1 (multiplet; NHCH 2 CH 3) and 8.89 (triplet, J 9 Hz; CH 2 CH 3).

60

Example 32 3-Ace toxymethyl-7β- [2- (2-chloroethyl) -carbamoyloxyimino-2-phenylacetamido7-ceph-3-em-4-carboxylic acid (synthetic acid)

To a solution of the 1.0 g cif syn isomer of 3-acetoxy-πιβΐγΙ-Τβ- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid ethyl acetate solvate in 21 ml of dime tylformamide containing 0.425 g of triethylamine was added 0.274 g of 2-chloroethyl isocyanate and the mixture was stirred at room temperature for one hour. The reaction mixture was acidified (2N hydrochloric acid) and extracted with ethyl acetate. The extracts were combined and extracted with saturated aqueous sodium bicarbonate. The combined aqueous extracts were acidified (2N hydrochloric acid) and extracted with ethyl acetate, dried, concentrated and added dropwise to petroleum ether with vigorous stirring. The precipitated solid was filtered, dissolved in 10 ml of ethyl acetate and added dropwise to 300 ml of petroleum ether. The precipitate was filtered and dried to give 0.76 g (74%) of the title acid, = + 28 ° (c = 1.0 in DMSO), \ max. (pH 6 phosphate buffer) 257.5 nm (ε = 20780), max. (nujol) 3270 (nh), 1780 (β-lactam), 1660 and 1520 cm -1 (amide), ^ values (DMSO-d6) include -0.10 (doublet, J 8 Hz; NH), 6.2 to 6.7 (multiplet; C-2 CH2 + -CH2CH2Cl), 7, S5 (singlet; OCCOCH3).

Example 33

Benzyl-2-phenylethylammonium-3-acetoxymethyl-7β- (2-hydroxy-imino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

To a solution of 0.084 g of sodium bicarbonate in 20 ml of water was added 0.507 g of 3-acetoxymethyl-7β- (2-hydroxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid ethyl acetate solvate and the resulting solution was mixed with a solution. of 0.396 g of benzyl-2-phenylethyl ammonium acetate in 10 ml of water. A white solid separated and after stirring for 15 min. filtered, washed with water and dried to 0.2 g (32%) of the title salt, Xmax. (pH 6 phosphate buffer) 254 nm (ε = 18000) max. (nujol) 1773 (β-lactam), 1650 and 1550 cm -1 (CONH), T (DMSO-d6) values include 0.4 (doublet, J 8 Hz; NH), 2.3 to 2.71 (multiplet aromatic protons), 7.99 (singlet; COCOCH 3), 143853 61

Example 34 3-Acetoxymethyl-7β-hydroxyimino-2- (thien-3-yl) -acetamido7-ceph-3-em-4-carboxylic acid, syn isomer

To an ice-cold solution of 1.59 g of t-butyl 1-3-ac ethoxymethyl 1-7β-aminoceph-3-em-4-carboxylate and 3.5 ml of propylene oxide in 15 ml of ethyl acetate was added with stirring a 1M solution of 5 , 3 ml of the syn isomer of 2-dichloroacetoxyaraino-2- (thien-3-yl) -acetyl chloride in ethyl acetate. After 1 1/2 hours at 20 ° C, the solution was washed with 2N HCl, saturated sodium bicarbonate solution and brine. The organic layer was dried and evaporated to give 2.8 g (100%) of the syn-isomer of t-butyl-3-acetoxymethyl-78- / 2-dichloroacetoxyimino-2- (thien-3-yl) -acetamido7-ceph -3-em-4-carboxylic acid as a white foam, ccD = + 47.0 ° (r - 0.9 in DiiSO), kmax (ethanol) 253 nm (ε = 13900), max (CHBr3) 3580 (OH), 3410 (NH), 1790 (β-lactam), 1728 (OCOCH ^) 1720 (C00-t-Bu) and 1700 and 1520 cm ”1 (CONH), C values (CDCl13) include 2.11 to 2.7 ( m; thienyl protons), 7.92 (s; OCOCH 3), 8.48 (s; t-Bu).

A solution of 2.7 g of the t-butyl ester in 25 ml of trifluoroacetic acid was left for 15 minutes, after which it was evaporated and the residue treated azeotropically with benzene. The oil was dissolved in ethyl acetate and extracted with saturated sodium bicarbonate solution. The aqueous solution was acidified to pH 1 with 2N HCl and extracted with ethyl acetate. The combined organic phases were dried and evaporated to a small volume which, upon addition to excess petroleum ether (bp 40-60 ° C) gave a white solid which was filtered and dried; there was obtained 1.5 g (60.3%) of the syn isomer of 3-acetoxymethyl-7β- / 5-hydroxyimino-2- (thien-3-yl) -acetamido7-ceph-3-em-4-carboxylic acid, aD = + 50.5 ° (c = 0.88 in DMSO), Xmax (phosphate buffer pH 6) 256.5 nm (ε = 18350), 0max (nujol) 3280 (NH and OH), 1770 (β-lactam) , 1726 (OCCOCH3), 1662 and 1522 (CONH) cnf1, δ values (DMSO-d6) include 0.36 (d; J 8 Hz; -NH-), 2.3 to 2.7 (m; tie-nyl) protons), 4.12 (dd; J 8 and 5 Hz; 7H), 4.77 (d, J 5 Hz; 6H), 7.96 (s; OCCOCH3).

Example 35 3-Acetoxymethyl-7β- / 2-hydroxyimino-2- (fur-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid, syn isomer

To an ice-cold solution of 1.76 g of t-butyl 3-acetoxymethyl-7β-aminoceph-3-em-4-carboxylate and 4 ml of propylene oxide in 15 ml of ethyl acetate was added with stirring a 1M solution of syn-2-dichloro -acetoxyimino-2- (fur-2-yl) -acetyl chloride in ethyl acetate (5.37 ml).

After one hour at 20 ° C, the solution was washed successively with 2N HCl, saturated sodium bicarbonate solution and brine. The organic layer was dried and evaporated to give the syn-isomer of the t-butyl ester as 3.04 g (100%) of a yellow foam, aD = + 66.5 ° (c = 0.9 in DMSO), Xrnax ( ethanol) 270 nm (ε = 16700); µmax (CHBr3) 3300 (NH), 1780 (β-lactam), 1726 (OCCOCH3 and C00-t-Bu) and 1680 and 1530 cm -1 (CONH), values ( CDCl 3) includes 2.31, 2.88 and 3.39 (multiples, furyl protons), 3.76 (s; CHCl 3), 7.92 (s; OCCOCH 3), 8.48 (s; t-Bu).

A solution of 2.94 g of the above syn isomer of t-butyl-acetoxymethyl-73- / 2-dichloroacetoxyimino-2- (gamma-2-yl) -acetamido7-ceph-3-em-4-carboxylate ml of trifluoroacetic acid was allowed to stand for 15 minutes after which the solvent was evaporated and the residue treated azeotropically with benzene. The oil was dissolved in ethyl acetate and extracted with saturated sodium bicarbonate solution. The aqueous solution was stained to pH 1 with 2N hydrochloric acid and extracted with ethyl acetate.

The combined organic phases were dried and evaporated to a small volume which upon addition to excess petroleum ether (bp 40-60 ° C) afforded a white solid which was filtered and dried; there was obtained 1.68 g of (76/0 3-acetoxymethyl-7β-, 5-hydroxyimino-2- (fur-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid, CCE = + 69 "( c = 1.2 in DMSO) max (pH 6 buffer) 269 nm (ε = 16300), -Omax (nujol) 1782 (β-lactam), 1728 (OCCOCH3), 1720 and 2600 (-C00H) and 1680 and 1536 cm -1 (CONH), <1 values (DMSO-d6) include 0.33 (d, J 8 Hz; NH), 2.19 and 3 »36 (2 s; furyl protons), 6.28 and 6.52 (2 doublets, branches of a quartet, J 18 Hz; C-2CH 2) and 7.95 (s; OCOCH 2).

Example 36 3-Acetylthiomethyl-7β-β-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em ~ 4 ~ carboxylic acid, syn isomer 1.03 g Diphenylme ty 1-3-acetylthiomethyl-7β-aminoceph-3-em-4-carboxylate hydrochloride was partitioned between 30 ml of ethyl acetate and 30 ml of aqueous sodium bicarbonate solution. The ethyl acetate layer was separated, washed with saturated sodium chloride solution, dried and filtered. The resulting solution was treated with a 1M solution of the syn isomer of 2-dichloroacetoxyimino-2- (thien-2-yl) -acetyl chloride in ethyl acetate (2.15 ml) and 0.5 ml of propylene oxide and stirred at 21 °. C for 40 minutes during which period a white precipitate appeared which redissolved. The solution was washed successively with saturated sodium bicarbonate solution, water and saturated sodium chloride solution and dried. Removal of the solvent in vacuo afforded 1.42 g of a dirt-white foam. This was dissolved in 2 ml of anisole and 8 ml of trifluoroacetic acid was added. After five minutes at 10 ° C, the reagents were removed in vacuo and the resulting gum was partitioned between ethyl acetate and aqueous sodium bicarbonate solution. The organic layer was re-extracted with sodium bicarbonate solution and the combined aqueous layers washed with ethyl acetate. The aqueous layer was adjusted to pH 2.0 with 2N hydrochloric acid and extracted with ethyl acetate. The extracts were washed with saturated sodium chloride solution and dried and the solvent removed in vacuo to give 0.97 g of the title compound as a light brown foam, aD = -52 ° (c = 1.04 in DMSO), Kmsac ( pH 6 buffer) 266 nm 64 143853 (ε = 13400), Omax (nujol) 2600, 1720 (COOH), 1770 (β-lactam), 1740 (SCOCHCH), 1660 and 1520 cm ”1 (CONH), C values ( DMSO-d6) includes 0.30 (d, J 9 Hz; NH), 2.40, 2.84 (m, thien-2-yl), 7.66 (s; SCOCH3).

The ester used as the starting material for the above acylation was obtained as follows.

a) Diphenylmethyl-3-acetylthiomethyl-7β-formamidoceph-3-em-4-carboxylate-1-oxide

A solution of 5.0 g of diethylmethyl-3-bromomethyl-7β-form amidoceph-3-em-4-carboxylate-ip-oxide in 30 ml of Ν, Ν-dimethylformamide was treated with 4.0 ml of ethanethiol acid and 1.4 ml of triethylamine and the reaction mixture was stirred for 16 hours at +19 ° C. The yellow solution was poured into 100 ml of water and 100 ml of ethyl acetate and the resulting insoluble solid was collected, washed with ethyl acetate and dried in vacuo over phosphorous toxide to give 2.422 g (49%) of diphenylmethyl-3-acetylthiomethyl-7β-formamidoceph. 3-em-4-carboxylate-1-oxide, m.p. 198-199 ° C (dec.), Otj = -60 ° (c = 1.0 in DMSO), max (ethanol) 273 nm (ε = 10300), Omax (CHBr 3) 3430 (NH), 1803 (β-lactam), 1725 (COOR), 1695, 1505 (CONH), 1042 cm -1 (S-O), <t (DMSO-d6) 1.66 ( d, J 9 Hz; NH), 1.78 (s; CHO), 2.59 (m; phenyl), 3.01 (s; CHPh 2), 7.68 (s; SCH 3).

b) Diphenylmethyl-3-acetylthiomethyl-7β-formamidoceph-3-em-4-carboxylate

A stirred solution of 1.00g diphenylmethyl-3-acetylthiomethyl "-7β-formamidoceph-3-em-4-carboxylate-iP" Oxide in 15 ml of N, N-dimethylformamide was cooled to -5 ° C and treated with 0.67 g of potassium iodide and 0.315 g of acetyl chloride The brown solution was stirred for five minutes at 0 ° C and then for 30 minutes after removal of the ice bath.

A solution of 0.60 g of sodium metabisulphite in 12 ml of water was added and the yellow suspension was stirred for five minutes at 20 ° C and then poured into 200 ml of water and extracted with dichloromethane. The combined extracts were washed with water and dried and the solvent removed in vacuo to give 0.92 g (94%) of diphenylmethyl-3- = acetylthiomethyl-7β-formamidoceph-3-em-4-carboxylate as a pale yellow powder, αβ = -136 ° (c = 1.0 in DMSO), λmax 268 nm (ethanol), (ε = 8950).

C) Di-phenylmethyl-3-acethyl-ethyl-7β-aminoc eph-3-em-4-carboxylate hydrochloride

A stirred suspension of 0.505 g of diphenylmethyl-3-acetyl-thiomethyl-7β- for mami doc eph-3-em-4-carboxy in 3 ml of methanol / ether 1: 1 at 0 ° C was treated for 10 minutes. with 0.468 g of phosphorus oxychloride. The ice bath was removed and stirring was continued for a further 20 minutes to give a clear solution. The reaction mixture was cooled to 0 ° C and a white crystalline precipitate appeared.

15 ml of ether was added and the precipitate was filtered off, washed with ether and dried in vacuo; there was obtained 0.456 g (89 $) of diphenylmethyl-3-acetylthiomethyl-7β-aminoceph-3-em-4-carboxyl at-hydrochloride as white microcrystals, m.p. 155-160 ° C (dec.), aD = -91 ° ( c = 0.83 in DMSO), Xmax (ethanol) 268 nm (e = 8700), Vmax (nujol) 1770 (β-lactam), 1720 (COOB), 1702 cnf1 (SCOCH +), values (DMS0-d6) includes 2.60 (m; Ph), 3.02 (s; CHPh 2), 7.66 (s; SCOCH 3).

Example 37 3-Crotonoyloxymethyl-7β-2-hydroxyimino-2- (thien-2-yl) acetamido7-ceph-3-em-4-carboxylic acid, syn isomer

A molar solution of the syn isomer of 2-dichloroacetoxy-imino-2- (thien-2-yl) -acetyl chloride in 2.15 ml of ethyl acetate was added to a solution of 0.933 g / 7 ethyl c eph-3-em-4-carboxylate in 15 ml of ethyl acetate containing 0.6 ml of propylene oxide.

After 30 minutes, the solution was washed with 2N hydrochloric acid, sodium bicarbonate solution, water and brine and dried and concentrated to 1.575 g of a foam.

A solution of 1.464 g of this foam in 2 ml of anisole was treated with trifluoroacetic acid at 20 ° C. After five minutes, the solution was concentrated. The oily residue was dissolved in ethyl acetate and the solution washed with water. The ethyl acetate solution was then extracted three times with dilute sodium bicarbonate solution. The combined extracts were washed with ethyl acetate and brought to pH 2 with concentrated hydrochloric acid. The solution is extracted with ethyl acetate and the extract washed with water and brine, dried and concentrated to a foam. Addition of ethyl acetate to this foam produced crystals which were filtered and washed with ethyl acetate; thus obtained the title oxime, aD = + 60 ° (c = 1 in DMSO), max (pH 6 phosphate buffer) 261 nm (ε = 16270), max (nujol) includes 1775 (β-lactam), 1733 (unsaturated ester), 1658 and 1545 (CONH) and 960 cm -1 (trans CH = CH), C values (DMSO-d6) include 0.26 (doubled, 8 Hz; NH), 2.38 and 2.83 ( 2 multiples; thienyl protons), 6.29 and 6.49 (doublets, J 18 Hz; C-2 H 2), 8.13 (doublets, J 7 and 1 Hz; -CH = CH-CH 3).

Example 38 7 P-2-Hydroxyimino-2- (thien-2-yl) -acetamido7-3- (1-phenyltetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylic acid, syn isomer

A solution of 2-dichloroacetoxyimino-2- (thien-2-yl) -acetyl chloride syn isomer in methylene chloride (7 ml of a 15% solution, ca.

3.5 mmol) was added over five minutes to a stirred solution of 1.779 g of diphenylmethyl-7β-amino-3- (1-phenylethetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylate hydrochloride and 1.5 ml of propylene oxide in 25 ml of dry methylene chloride. After stirring for an additional 20 minutes at 23 ° C, the solution was washed successively with saturated sodium bicarbonate, water and brine (25 ml of each) and dried and evaporated to 2.62 g of a foam. A solution of this foam in 10 ml of benzene was purified by chromatography on silica gel with benzene / stylet acetate 10: 1 as eluent. Appropriate fractions were combined and evaporated to dryness in vacuo to give 1.27 g of diphenylmethyl-7β-T2-hydroxyimino-2- (thien-2-yl) -acetamido-3- (1-phenyltetrazol-5-ylthiomethyl) -ceph -3-em-4-carboxylate as a foam. A solution of this foam in 5 ml of trifluoroacetic acid and 1 ml of anisole was kept at 23 ° C for five minutes and the solvent was removed in vacuo at 40 ° C. The residue was dissolved in 50 ml of ethyl acetate, the solution was extracted with sodium bicarbonate solution and the alkaline extract was covered with 100 ml of ethyl acetate and acidified to pH 2 with 2N hydrochloric acid. The organic phase was separated and washed with water and brine, then dried and evaporated to 750 mg of a foam. A solution of this foam in ethyl acetate was added to petroleum ether to give 469 mg (29%) of the title acid as a pale yellow solid, ocD = -51.4 ° (c = 0.91 in acetone), max (pH 6 phosphate buffer) 264 nm (ε = 19500), i) max (nujol) 3320 (NH), 1784 (β-lactam), 1722 67 143853 (COOH), and 1674 and 1530 cm -1 (CONH), ( DMSO-d6) 0.26 (NH, doublet, J 8 Hz) 2.31 (s; Ph), 4.13 (C 7 -H, double doublet, J 5 and 8 Hz) 4.82 (C 6 H, doublet, J 5 Hz), 5.39 and 5.73 (quartet, J 13 Hzj C-3 CH 2) and 6.28 (s; C-2 CH 2),

The amino ester used as the starting material for the above-described acylation was prepared in a similar manner to the amino ester described in Example 46, replacing the 5-mercapto-2-phenyl-1,3,4-oxadiazole with 5-mercapto-1 phenyltetrazole to give diphenylmethyl-7β-amino-3- (1-phenyltetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylate hydrochloride, kmax (ethanol) 251.5 nm (ε = 10000), %) max (CHBr3) 1782 (β-lactam) and 1720 cm "1 (C00R), C values (DMSO-d6) include 2.32 (Ph-N)), 4.73 (C-6H and C-7H) ), 5.32 and 5.68 (C-3 CH 2, quartet, J 14 Hz) and 6.10 (C-2 CH 2).

Example 39 7 β-2-Hydroxyimino-2- (thien-2-yl) -acetamido7-3- (1-methyltetrazole-5-ylthiomethyl) -ceph-3-em-4-carboxylic acid, syn isomer

A solution of the syn isomer of 2-dichloroacetoxyimino-2- (thien-2-yl) acetyl chloride in dry methylene dichloride (6 ml of 15% solution, about 3 mmol) was added over a period of five minutes to a solution of 1 24 g of diethylmethyl-7β-amino-3- (1 * -methyltetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylate and 0.8 ml of propylene oxide in 20 ml of methylene dichloride. After stirring for 30 minutes at 23 ° C, aqueous sodium bicarbonate was added and the biphasic mixture was stirred for a further 30 minutes. The organic phase was separated, washed with brine and dried and evaporated to 2.0 g of a foam. A solution of this foam in 20 ml of benzene was purified by chromatography on silica gel with benzene / ethyl acetate 5: 1 as eluent. Appropriate fractions were combined and evaporated in vacuo to give 1.68 g of diphenylmethyl-7β- / 2-hydroxyimino 2- (thien-2-yl) acetamido7-3- (1-methyltetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylate as a foam. A solution of this foam in 6 ml of trifluoroacetic acid and 1.5 ml of anisole was kept at 23 ° C for five minutes and the solvents were removed at 40 ° C / 2 mm Hg. The residue was dissolved in 30 ml of ethyl acetate and the solution was extracted with sodium bicarbonate solution. The alkaline extract was covered with ethyl acetate and acidified to pH 2 with 2N hydrochloric acid. The organic phase was separated and washed with water and brine and the solvent was evaporated in vacuo. The residue was dissolved in acetone and the solution treated with some charcoal and filtered through a diatomaceous earth mat; the filtrate was dried and evaporated to 1.085 g of a foam. A solution of this foam in ethyl acetate was transferred to petroleum ether to give the title acid as a colorless amorphous solid in an amount of 852 mg (71%), and = -90 ° (c = 0.91 in acetone), µmax (pH 6 phosphate buffer) 267 »5 nm (ε = 18000), Omax (nujol) 3290 (NH), 1780 (β-lactam), 1720 (COOH) and 1670 and 1530 cm cm ((CONH), DMSO-d6) values include 0.25 (NH, doublet, J 8 Hz) 4.14 (C-7H, double doublet, J 4.5 and 8 Hz) 4.79 (C-6H, doublet, J 4, 5 Hz), 6.05 (CHg).

Similarly prepared:

Example 40 7β - Hydroxyimino-2- (fur-2-yl) -acetamido7-3- (1-methyltetrazole-5-ylthiomethyl) -ceph-3-em-4-carboxylic acid, syn isomer, aD = -102 ° (c = 0.9 in acetone), kmax (pH 6 phosphate buffer) 273.5 nm (ε = 20700), λ max (nujol) 3280 (NH), 1780 (β-lactam), 1720 (COOH) and 1674 and 1530 cm -1 (CONH), <C values (DMSO-d6) include 0.33 (NH, doublet, J 8 Hz) and 6.07 (CH 3).

The starting ester of the acylations of Examples 39 and 40 was prepared by the following: a) Diphenylmethyl-3- (1-methyletrazol-5-ylthiomethyl) -7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate

A solution of 17.505 g of diphenylmethyl-3-bromomethyl-7β- (2-thienylacetamido) -ceph-3-em-4-carboxylate and 3.486 g of 5-mercapto-1-methyltetrazole in 250 ml of dry tetrahydrofuran was stirred at 10 ° C. over five minutes with a solution of 4.16 ml of triethylamine in 50 ml of dry tetrahydrofuran. The resulting suspension was stirred for an additional 45 minutes during which time the temperature was allowed to rise to ca. To a stirring mixture of 50 ml of 2N hydrochloric acid, 700 ml of water, 700 ml of brine and 1.5 l of ethyl acetate. The organic phase was separated, washed with 2N hydrochloric acid, water and brine and treated with some charcoal and filtered through a diatomaceous earth mat. The filtrate was dried and evaporated to 20.6 g of a foam. This foam was dissolved in 200 ml of warm methanol and the solution cooled to give the ester as colorless plates in an amount of 14.391 g (77.5%), kmax (ethanol) 264 nm (ε = 8700), Omax (CHBr3) 3420 (NH) , 1780 (β-lactam), 1710 (COOR) and 1680 and 1502 cm cm 1 (CONH), δ values (CDCl ^) include 3.40 (NH, doublet, J 9 Hz), (C-7H, double doublet, J 5 and 9 Hz) 5.02 (C-6H, doublet, J 5 Hz), 6.19 (CH 3).

b) Diphenylmethyl-7β-amino-3- (1-methyltetrazol-5-ylthiomethyl) -ceph-3-em-4-carboxylate

A suspension of 416 mg of phosphorus pentachloride in 10 ml of dry methylene dichloride was stirred at 23 ° C and a solution of 0.16 ml of pyridine was added. The hot suspension was stirred for 10 minutes and cooled to 0 ° C, then treated for 10 minutes with a solution of 619 mg of diphenylmethyl-3- (1-methyltetrazol-5-ylthiomethyl) -78- (2-thienylacetamido) -ceph. 3-em-4-4: arboxylate in 15 ml of methyl endichloride. After stirring for a further 45 minutes, during which time the temperature was allowed to rise to 23 ° C, the solution was added over a period of five minutes to a stirred and cooled mixture of 5 ml of methanol and 15 ml of methylene dichloride. The resulting solution was stirred for a further 10 minutes, washed with 20 ml of N hydrochloric acid and stirred for 30 minutes with 50 ml of saturated sodium bicarbonate solution. The organic phase was separated, washed with 25 ml of saturated aqueous sodium bicarbonate and treated with some charcoal. The mixture was filtered through a diatomaceous earth mat and the filtrate was evaporated to 640 mg of a gum triturated with ether (20 ml) to give the amine as 403 mg (81.5%) dirt white solid, Kmax (CHCl3) 268.5 nm (ε = 7700 ), Vmax (CHBr3) 3416 and 3350 (NH2), 1780 (β-lactam) and 1720 cm cm 1 (C00R), 7 values (CDCl13) include 5.00 and 5.15 (C-6H and C-7H, two doublets, J 5 Hz), 6.14 (CH 3 -) and 7.59 (NH 2).

Example 41 Similar to Example 39, acylation of di-phenylmethyl-7β-amino-3- (5-methyl-1,3,4-thiadiazolyl-2) -thiomethylceph-3-em-4-carboxylate with syn -2-dichloroacetoxyimino-2- (fur-2-yl) -acetyl chloride and subsequent removal of the protection syn isomer of 70 U38S3 7P - / - - hydroxyimino ~ 2- (fur-2-yl) -acetamido7-3- (5-methyl-1,3,4-thia-diazol-2-yl) -thiomethylcepli-3-em-4-carboxylic acid, ε C values (DMSO-d6) include 0.36 (d, J 9 Hz, NH), 2.23, 3.39 (25, furyl protons), 4.19 (dd, J 5 and 9 Hz, C-7H), 4.83 (d, J 5 Hz, C-6H), 5 , 45, 5.90 (q, J 14 Hz, C-3 CH 2), 6.21, 6.44 (q, J 18 Hz, C-2 H2), 7.33 (s, CH 2).

Example 42 7 β-2-Hydroxyimino-2- (thien-2-yl) -acetamido [3- (5-phenyl-1,3,4-oxadiazol-2-ylthiomethyl) -ceph-3-em-4 -carboxylic acid, syn-isomer

A solution of the syn isomer of 2-dichloroacetoxyimino-2- (thien-2-yl) -acetyl chloride in methylene dichloride (7 ml 15% s solution) was added over 5 minutes to a stirring solution of 1.779 g diphenylmethyl-78 -amino-3- (5-phenyl-1,3,4-oxadiazol-2-ylthiomethyl) -ceph-3-em-4-carboxylate hydrochloride and 1.5 ml of propylene oxide in 25 ml of dry methylene dichloride. After stirring for a further 30 minutes, the solution was quenched with saturated aqueous sodium bicarbonate, water and brine (25 ml each) and dried and evaporated to 2.34 g of a slum. A solution of this foam in 10 ml of benzene was purified by chromatography on silica gel with benzene / ethyl acetate 10: 1 as eluent. Appropriate fractions were combined and evaporated to dryness in vacuo and the residue was triturated with ether (25 mL) to give 370 mg (41%) of diphenylmethyl-7β-75-hydroxyimino-2- (thien-2-yl) -acetamido-3- (5) -phenyl-1,3,4-oxadiazol-2-ylthiomethyl) -ceph-3-em-4-carboxylate as colorless prisms. A suspension of 810 mg of this ester in a mixture of 3 ml of trifluoroacetic acid and 0.8 ml of anisole was shaken at 23 ° C until all the solid had dissolved, which was about 10 minutes. The reagents were evaporated at 40 ° C / 2 mm Hg and the residue dissolved in ethyl acetate. The solution was extracted with aqueous sodium bicarbonate and the alkaline extract was covered with ethyl acetate and acidified to pH 2 with 2N hydrochloric acid. The organic phase was separated and washed with water and brine, then the solvent was evaporated in vacuo. The residue was dissolved in acetone and then the solution was treated with some charcoal and filtered through a diatomaceous earth mat. The filtrate was dried and evaporated to 470 mg of a foam. A solution of this foam in ethyl 71 was transferred to petroleum ether to give 410 mg (66%) of the title acid as a colorless amorphous solid, = -118 ° (c = 1.08 in acetone), kmax (pH 6 phosphate buffer) 274 nm (ε = 28800), Omax (nujol) 3280 (NH), 1786 (β-lactam), 1720 (COOH) and 1672 and 1530 cm cm ”(CONH), <Tverages (DMS0- d6) includes 0.24 (NH, doublet, J 8 Hz) 4.15 (C-7H, double doublet, J 4.5 and 8 Hz), 4.78 (C-6H, doublet, J 4.5 Hz) ).

The amino ester used for the acylation just described was prepared as follows:

Diphenylmethyl-73-amino-3- (5-phenyl-1,3,4-oxadiazol-2-ylthiomethyl) -ceph-3-em-4-carboxylate hydrochloride

A solution of 10.07 g of diphenylmethyl-3-bromomethyl-7β-formamidoceph-3-ent-4-carboxylate-1-oxide and 3,564 g of 5-mercapto-2-phenyl-1,3,4-oxadiazole in 250 ml of dry Ν, Ν-dimethylformamide was stirred at 0 ° C and treated over 10 minutes with a solution of 3.2 ml of triethylamine in 20 ml of Ν, Ν-dimethylformamide. The resulting solution was stirred for an additional 2 1/2 hours during which time the temperature was allowed to rise to ca. 20 ° C, then the solvent was evaporated at 40 ° C in vacuo. The residue was partitioned between 250 methylene dichloride and 150 ml of water and the organic phase was separated, washed with 2N hydrochloric acid, water and brine and dried, then the solvent was evaporated to yield 12.076 g of diphenylmethyl-7β-formamido-3- (5-phenyl-1,3 (4 (oxadiazo-2-ylthiomethyl) -ceph-3-em-4-carboxylate-1-oxide as an orange solid. A suspension of this material in 250 ml of dry methylene dichloride was stirred at -20 ° C and treated over 10 minutes with a solution of 5.7 ml of phosphorus tribromide in 25 ml of methylene dichloride.

The resulting solution was stirred for an additional hour at -10 to -15 ° C and then added over 10 minutes to a stirred and cooled mixture of methylene dichloride and saturated sodium bicarbonate solution. The two-phase system was stirred for an additional hour at ca. 23 ° C; the organic phase was separated and washed with saturated sodium bicarbonate solution and brine and treated with some charcoal. The suspension was filtered and the filtrate dried and evaporated in vacuo to give 11.231 g of diphenylmethyl-7β-formamido-3- (5-phenyl-1,3,4-oxadiazol-2-ylthiomethyl) -ceph-3-em-4-carboxylate like a foam. A suspension of this foam in a mixture of 100 ml of methanol and 100 ml of ether was stirred at 0 ° C and treated with 3.4 ml of phosphorus oxychloride over 20 minutes. After stirring for an additional three hours, the suspension was poured into 200 ml of ether to give the title hydrochloride as 8.707 g (77%) of a light brown solid. λ max (ethanol) 272 nm (ε = 21200), λ max (nujol) 1790 (β-lactam) and 1720 cm ”1 (C00R), lti values (DMSO-d6) include 4.70 (C-6H and C-7H), 5.43 and 5.65 (C-3 CH2, quartet, J 13 Hz).

Example 43 3-Acetoxymethyl-7β- / 2-ethoxycarbonyloxyimino-2- (thien-2-yl) acetamido7-ceph-3-em-4-carboxylic acid, syn isomer

To a solution of 0.6 g of the syn isomer of sodium 3-acetoxymethyl-7β-7Sydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate in 25 ml of saturated aqueous of sodium bicarbonate solution, a solution of 1.5 ml of ethyl chloroformate in 20 ml of ethyl acetate was added and the whole was shaken vigorously for one minute. The bicarbonate phase was separated and shaken with an additional portion of 1.0 ml of ethyl chloroformate in 20 ml of ethyl acetate. The bicarbonate phase was separated, washed with ethyl acetate, acidified and extracted with ethyl acetate. The combined extracts were dried and concentrated to ca. 5 ml. The concentrate was added dropwise and with stirring to 200 ml of petroleum ether (boiling point 40-60 ° C). The precipitated solid was filtered and dried to give 0.47 g of the title acid, αβ = + 42 ° (c = 0.9 in DMSO), kmax (pH 6 phosphate buffer) 265 nm (ε = 16900), Omax (nujol) 3290 (NH), 1784 (β-lactam), 1740 (acetate), 1690 and 1550 cm cm 1 (CONH), δ values (DMSO-d NH), 2.08, 2.53, 2.74 (three quartets; thienyl protons), 5.70 (q, J 8 Hz; -CHpCH 2, 7.96 (s; OAc), 8.72 (t, J 8 Hz; -CH 2 CH 3).

Example 44 7 β-2-Acetoxyimino-2- (thien-2-yl) -acetamido-3-acetoxymethylceph-3-em-4-carboxylic acid, syn isomer

A solution of 0.7 g of the syn isomer of sodium 3-acetoxymethyl-7β- / 2-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4- 73 1438 * 3 carboxylate in 20 ral water was stirred with 50 ml of ethyl acetate and the pH of the aqueous layer was adjusted to 1 with 2N hydrochloric acid. The ethyl acetate phase was separated, combined with an ethyl acetate extraction of the aqueous phase, dried and evaporated to an oil. The oil was dissolved in 28 ml of ethyl acetate containing 2.1 g of pyridine and treated dropwise with stirring at room temperature with a solution of 1.4 g of acetyl chloride in 14 ml of ethyl acetate. After stirring for 12 minutes, the mixture was poured into 2N hydrochloric acid and the ethyl acetate lid was separated, washed with water and extracted with saturated aqueous sodium bicarbonate solution. The combined extracts were washed with ethyl acetate, acidified with 2N hydrochloric acid and extracted three times with ethyl acetate.

The combined organic extracts were dried, concentrated to 5 ml and added dropwise to 250 ml of petroleum ether (bp 40-60 ° C).

The precipitated solid was filtered and dried to give 0.5 g (68%) of the title acid, max (pH 6 phosphate buffer) 265 nm (ε = 15500), max max (nujol) 3270 (NH) , 1780 (β-lactam), 1736 (acetate), 1680 and 1540 cm cm 1 (CONH), C values (DMSO-d6) include -0.15 (d, J 9 Hz; NH), 2.08, 2 , 54, 2.73 (ten-2-yl protons), 7.79 (s; = NOCOCH3), 7.94 (singlet, OCCOCH3).

Example 45 3-Aceticloxy 7-β - [(2- (2-chloroethylcarbamoyloxyimino) -2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid, syn isomer

A solution of 0.89 g of the syn isomer of sodium 3-acetoxymethyl-7β- / 2-hydroxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylate in 11 ml dimethyl foramide containing 0.22 g of triethylamine was treated with 0.25 g of 2-chloroethyl isocyanate. The solution was allowed to stand at room temperature for 30 minutes and then at 5 ° C for an additional hour. It was poured into cold (5 ° C) 2N hydrochloric acid and extracted three times with ethyl acetate, then the combined extracts were washed with 2N hydrochloric acid and then water and extracted three times with cold (5 ° C) saturated aqueous sodium bicarbonate solution. The combined aqueous extracts were washed with ethyl acetate, acidified with 2N hydrochloric acid and rapidly extracted with ethyl acetate. The combined extracts were dried and concentrated to ca. 5 ml and the concentrate was added dropwise to 250 ml petroleum ether (boiling point 40-60 ° C).

74 163853

The precipitated solid was filtered and dried to give 0.8 g (76%) of the title acid, = + 33 ° (c = 0.7 in DMSO), max (pH 6 phosphate buffer) 264 nm ( ε = 17200), Omax (nujol) 1786 (β-lactam), 1734 (acetate), 1682 and 1508 cm -1 (CONH),

C values (DMSO-d6) include -0.12 (doublet, J 9 Hz; NH), 2.12, 2.56, 2.74 (three quartets; thienyl protons), 7.97 (s; OCOCH 3).

Example 46 β- / 7β- (2-Hydroxyimino-2- (fur-2-yl) -acetamido) -ceph-3-em-3-ylmethyl7-41-carbamoylpyridinium-4-carboxylate, syn isomer

A solution of 2.5 g of 3-acetoxymethyl-7β-T ^-hydroxyimino- (fur-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid and 2.5 g of isonicotinamide in water heated to 65 -70 ° C in nitrogen for 3 1/2 hours. The cooled mixture was passed through a column of AGlx8 resin (acetate form) and eluted with water. Fractions were combined on the basis of electrophoreographs and freeze-dried to a pore which was well washed with acetone and dried to give 0.834 g (23%) of the title compound, αβ = -61 ° (c = 0.84 in DMSO), max (pH 6 buffer) 267.5 nm (ε = 19500), Omax (nujol) 3340 (NH), 1772 (β-lactam), 1680 and 1560 (CONH), 1608 cm -1 (COO ) values (d6-DMSO) include 0.40, 1.26, 1.50 and 1.75 (multiplets, aromatic protons), 1.26 and 1.75 (broad singlets; CONHg), 4.26 ( multiplet; C-7H), 4.28 and 4.77 (C-3 CH2), 4.88 (d, J 5 Hz; 0-6H).

Example 47 3-Acetoxymethyl-7β- / 2-hydroxyimino-2- (pyrid-4-yl) -acetamido7-ceph-3-em-4-carboxylic acid trifluoroacetic acid salt, syn isomer a) t-Butyl-3-acetoxymethyl acid 7β- (pyrid-4-ylacetamido) -ceph-3-em-4-carboxylate

A suspension of 6.0 g of pyrid-4-ylacetyl chloride hydrochloride in 50 ml of ethyl acetate containing 12 ml of propylene oxide was stirred, cooled in ice and treated dropwise with a solution of 5 »0 g of t-butyl-3-acetoxymethyl ether. 7β-Aminoceph-3-em-4-carboxylate in 50 ml of ethyl acetate. The mixture was stirred at 20 ° C for 20 hours and then washed with saturated sodium bicarbonate solution and extracted with 2N hydrochloric acid. The aqueous extract was washed with ethyl acetate, neutralized with saturated sodium bicarbonate solution, and extracted with ethyl acetate. The extract was washed with water, dried over magnesium sulfate and evaporated to a small volume, after which crystallization began.

The crystalline solid was well washed with petroleum ether (boiling point 40-60 ° C) and dried to give 5.4 g (80 µg of the amine) of the title ester as a brownish yellow powder. A 250 mg portion of this was recrystallized from ethyl acetate to 170 mg white needles, α D = + 94 ° (c = 0.8 in DMSO), λ max (ethanol) 256.5 nm (ε = 9900),> max CHBr ^) 3412 (NH), 1784 (β-lactam), 1736 (OAc), 1722 (C00R), 1690 and 1512 cm'1 (CONH), t (d6-DMSO) 0.81 (d, J 8 Hz ; NH), i, 47 and 2.68 (2d, J 5 Hz; aromatic protons), 4.27 (q, J 8 and 5 Hz; C-7H), 4.85 (d, J 5 Hz; C-6H), 5.01 and 5.36 (2d, J 13 Hz; C-3 CH 3), 6.38 (s; CH 2 CONH), 6.39 (s; C-2 CH 3), 7.96 (s; COCOCH 3) and 8.50 (s; C (CH 3) 3).

b) t-Butyl 3-acetoxymethyl-7β- [5-hydroxyimino- (pyrid-4-yl) -acetamido] -ceph-3-em-4-carboxylate

A solution of 3.0 g of t-butyl 3-acetoxymethyl-7β- (pyrid-4-ylacetamido) -ceph-3-em-4-carboxylate in 30 ml of acetic acid was stirred, cooled briefly in ice, and treated dropwise over time. 3-4 minutes with a solution of 1.38 g of sodium nitrite in 10 ml of water. The mixture was stirred at 20 ° C for 30 minutes and then diluted with water and extracted with ethyl acetate. The organic extract was washed with saturated sodium bicarbonate solution and water and then dried over magnesium sulfate. Evaporation almost to dryness gave a residue which was dissolved in the least amount of acetone and added dropwise to stirred petroleum ether (boiling point 40-60 ° C). The resulting solid was collected, washed with petroleum ether and dried, yielding the title oxime as 2.9 g (91%) of a white powder, which was shown to be the syn / anti-mixture 30:70 by nuclear magnetic resonance spectrum.

2.5 g of the oxime mixture was chromatographed on silica gel and eluted with methanol / chloroform 1:49 to give 1.09 g of the anti-isomer of the title compound, corresponding to a yield of 44% from the mixture.

Further elution with methanol / chloroform 1:49 gave material which was dissolved in acetone and added to stirred petroleum ether (bp 40-60 ° C) to give 0.81 g of syn isomer one of t-butyl-3-acetoxymethyl 7 β-2-Hydroxyimino- (pyrid-4-yl) - ac etamido7-ceph-3-em-4-carboxylate, corresponding to a yield of 32% from the mixture, ocD = + 57 ° (c = 1.11 in DMSO), max (ethanol) 253.5 nm (ε = 17600), µmax (nujol) 3220 (NH), 1784 (β-lactam), 1744 (OAc), 1710 (COOR), 1640 and 1526 cm -1 (CONH), δ (d6-DMSO) -2.26 (s; NOH), 0.27 (d, J 8 Hz; NH), 1.33 and 2.47 (2d, aromatic protons ), 4.04 (q, J 5 and 8 Hz; C-7H), 4.71 (d, J 5 Hz; C-6H), 4.98 and 5.33 (2d, J 13 Hz; C- 3 CH 2), 6.20 and 6.53 (2d, J 18 Hz; C-2 CH 2), 7.94 (s, 0 COCH 3), 8.49 (s; C (CH 3) 3).

c) 0.6 g of the syn-t-butyl ester in 15 ml of trifluoroacetic acid was left at 20 ° C for 15 minutes and then evaporated to dryness. The residue was treated with acetone and the solvent removed in vacuo, and this process was repeated. The residue in acetone was slowly added to petroleum ether (boiling point 40-60 ° C).

The precipitate was collected and dried to give 0.66 g (99%) of the syn isomer of 3-acetocymethyl-7β- / 2-hydroxyimino-2- (pyrid-4-yl) -acetamido7-ceph-3-em. 4-carboxylic acid trifluoroacetic acid salt, αβ = + 47.5 ° (c = 0.84 in DMSO), λ max (pH 6 buffer) 248.5 nm (ε = 17000),% max (nujol) 3250 ( NH), 1772 (β-lactam) 1720 (OAc), 1660 (CF3C00 "), 1660 and 1530 cm -1 (COHH), * C (d6-DMSO) 0.16 (d, J 8 Hz; NH), 1.14 and 2.20 (multiples, aromatic protons), 4.06 (q, J 5 and 8 Hz; C-7H), 4.71 (d, J 5 Hz; C-6H), 4.92 and 5.24 (2d, J 13 Hz; C-CH 2), 6.25 and 6.61 (2d, J 18 Hz; C-2 CH 2), 7.93 (s; ococh 3).

Claims (3)

143853 77 1. Analogous Process for the Preparation of Cephalosporin Compounds of the General Formula sv R.C.CONH__S '> in Zj. Wherein R represents phenyl, halogen substituted phenyl, naphthyl or a heterocyclic group having 5 or 6 ring atoms, at least g of which is S, N or O, R is a hydrogen atom, optionally halogenated alkanoyl group having up to 7 carbon atoms, an alkoxycarbonyl group having up to 7 carbon atoms, a benzoyl or nitrobenzoyl group or an alkylcarbamoyl or haloalkylcarbamoyl group having 1-7 carbon atoms in the alkyl moiety and RX a methyl or vinyl group or a group -CH2Y where Y represents (i) , carbamoylpyridinium or the residue of a heterocyclic amine having 1-4 heteroatoms with at least one hetero atom being nitrogen, (ii) azido, CC (iii) a group -SR wherein R is C 1-4 alkyl, C 2-6 alkanoyl or a 5- or 6-membered heterocyclic group containing at least one of the atoms 0, N and S, (iv) a group -0Rt where Rfc is hydrogen or alkyl, or 9 (v) a group -0.C0R where R is C1-4 alkyl, C2_4 alkenyl or phenyl, or non-toxic salts, or hydrates thereof f) characterized in that a) condensing a compound, which may be in the form of an acid addition salt, of the general formula H2N-I-J_N 1 -R * 111 cr ^ COOR1 X where R has the meaning given above B represents> S or> S-> 0, R 2 represents hydrogen or a carboxyl blocking group and the broken line between positions 2, 3 and 4 means that the compound may be a ceph-2-em or ceph-3-em compound , with an acid of the general formula RCCOOH 'T II VI N SVvORa where R and R a have the above meanings, or an acid of the general formula RCCOOH blocking group, or a reactive derivative of one of these acids, or b) reacting a compound of the general formula B RCCO.NH --- S \ II! N \ c / -N \ J-CH2Y '11 X0RC / 2 o In COOR where B, R1 and the broken line have the above meanings C, R is hydrogen or the group R as defined above and Y' is a chlorine, bromine or iodine atom or an acetoxy group, having a nucleophile corresponding to group Y, 79 U3853 and then, in a formed compound, if necessary, one or more of the following reactions (i) convert a group of RCCO 4 in a manner known per se, 2 3 (ii) converts an Δ isomer to the desired Δ isomer in a manner known per se, (iii) removes any blocking groups present, (iv) reduces a compound where B represents > S + 0 to form the desired compound where B represents> S in a manner known per se, (v) reacting a compound wherein Y is azido with a dipole hydrophilic to form a compound of formula I wherein Y is another of the groups defined above Y, (vi) deacylates in a manner known per se a compound wherein Y is e n acyloxy group to form a 3-hydroxymethyl group and / or (vii) acylating in a known manner a compound wherein Y is a hydroxy group to form a 3-acyloxymethyl compound and recovering the desired compound if desired of formula I or a non-toxic salt or hydrate thereof. Process according to claim 1, characterized in that it is used for the preparation of compounds of formula S RU. C. CONH ____y '\ N f B XOH ^ -Nx ^ 1-CH2OCO-CH3 COOH wherein RU represents phenyl, halogen-substituted phenyl, thienyl, naphthyl or furyl, or non-toxic salts or hydrates thereof. Process according to claim 1, characterized in