DK147683B - METHOD OF ANALOGY FOR THE PREPARATION OF CEPHALOSPORINE COMPOUNDS OR TOXIC SALTS, BIOLOGICAL ACCEPTABLE ESTERS, 1-OXYDS OR SOLVATES THEREOF - Google Patents

Description

147683

The present invention relates to an analogous process for the preparation of cephalosporin compounds, in particular to certain 7- (2-oxyiminoethers) having vision configuration or wholly dominant (at least 90%) vision configuration and the general formula I of claim 1, wherein R 1 and R have the same meanings stated, or non-toxic salts, biologically acceptable esters, 1-oxides or solvates thereof. The process according to the invention is characterized by the characterizing part of claim 1.

The cephalosporin compounds of this specification are named from the substance cepham in accordance with J. Am.Chem. Soc. 1962, 84, 3400, and the name cephem refers to the basic cepham structure with one double bond.

Many cephalosporin compounds with some degree of antibacterial activity are known and these compounds have 3 Δ unsaturation and are usually substituted at the 3 position with a methyl group or substituted methyl group and at the 73 position with an acylamido group. It is now generally agreed that the antibiotic properties of a given ceph-3-em-4-carboxylic acid are predominantly controlled by the nature of both the 7S-acylamido group therein and the substituent at the 3-position; Comprehensive studies have been conducted to find combinations of such groups that will provide antibiotics with special properties.

Cephalosporin antibiotics are extensively used to treat human and animal diseases caused by pathogenic bacteria, for example, to treat diseases caused by bacteria that are resistant to other antibiotics such as penicillin compounds, and to treat penicillin-sensitive patients. For many applications, it is desirable to use a cephalosporin antibiotic that exhibits activity both against Gram-positive and Gram-negative microorganisms, and a major research effort is directed towards the development of improved broad-spectrum cephalosporin antibiotics.

The practical utility of a considerable number of known commercial and experimental cephalosporin antibiotics is limited by their relatively high sensitivity to 3-lactamases formed by many bacteria. Therefore, a desirable property of a broad-spectrum cephalosporin antibiotic is that it exhibits substantial resistance to β-lactamases, including those produced by gram-negative bacteria.

A further difficulty of many cephalosporin antibiotics for therapeutic use is that they can undergo degradation in vivo. Thus, it has been found that a significant number of known cephalosporin antibiotics have the disadvantage of being deactivated upon administration, often rapidly, of enzymes such as esterases present in the body.

Among known related cephalosporin compounds, in particular, there are three groups known from DK patent applications Nos. 2373/73, 381/72 and 382/72.

Those from DK ans. 2373/72 known compounds have the general formula R.C.CO.NH-- 1 I] "-OR * / -

Wherein R represents phenyl, naphthyl, thienyl, furyl, benzothienyl, benzofuryl, pyridyl or N-methylpyrrolyl, Ra represents a C 1-4 alkyl, C 2-8 alkenyl, propinyl, propargyl, C 3-6 cycloalkyl, phenyl, phenyl-C 1-4 alkyl, triphenylmethyl, furylmethyl or pyridylmethyl group or any of these groups substituted with one substituent in the form of hydroxy, methoxy, ethoxy, bromo, nitro, amino, carboxy or benzoyloxy, X represents methyl, vinyl or the group -CH 2 Y where Y can have a wide range of meanings, the compounds being syn-isomers.

Those from DK ans. 381/72 known compounds have the general formula

Ry. ^. CONH ---

N JT "X

N. w Å N · 1 R X

0Rb /

COOH

wherein R 1 represents phenyl, halogen substituted phenyl, naphthyl, pyridyl, thienyl or furyl, R a hydrogen atom, optionally halogenated alkanoyl group of up to 7 carbon atoms, an alkoxycarbonyl group of up to 7 carbon atoms, a benzoyl or nitrobenzoyl group or an alkylcarbamoyl group or an alkylcarbamoyl group 1-7 carbon atoms in the alkyl moiety and Rx a methyl or vinyl group or a group -CH ~ Y where Y can have a wide variety of

^ X

meanings, but not so that R may be a carbamoyloxy methyl group; also these compounds are available as syn isomers or predominantly syn isomers. Those from DK ans. 382/72 are the anti-isomers corresponding to the compounds of formula IX.

As a result of prolonged studies on numerous cephalosporin compounds, we have now succeeded in finding a group of cephalosporin antibiotics with a particular combination of 7β-acylamido group and substituent at the 3-position, which gives the compound good broad-spectrum activity coupled with the above desirable properties with respect to the above. high stability to β-lactamase and good in vivo stability. These compounds are characterized in that the 7β-acylamido group is a 2-aryl-2- (etherified oxyimino) -acetamido group which is substantially in syn configuration (as defined later in this specification) and that the 3-substituent is a carbamoyloxymethyl group. As mentioned, the compounds have the formula I shown in the preamble of claim 1, and the meanings of R and R are defined together.

Thus, the compounds must have the isomeric syn form 2 (cis form) with respect to the configuration of the group OR relative to the carboxamido group. In this specification, the vision configuration is structurally specified as follows: R1.C.CONH- ft

N

"^ OR2

The syn form is attributed to these compounds on the basis of a work by Ahmad and Spenser reported in Can. J. Chem. 1961, 39, 1340.

The term non-toxic about the derivatives of the compounds of the invention is intended for such derivatives which are physiologically acceptable at the doses in which they are administered. Thus, such derivatives are salts, biologically acceptable esters, 1-oxides or solvates (especially hydrates) of the compounds of the invention.

147683 4

The biological effect of a number of the compounds prepared by the process according to the invention is in various ways compared to known compounds, both those known from the aforementioned three Danish patent applications and others.

First, reference is made to Table A, showing the in vitro effect of the majority of the compounds listed in the examples below against some representative test organisms measured by the agar tube dilution technique. The table indicates MIC (least inhibitory concentration) in µg / ml, using the test organisms at a concentration of 10 per ml. ml. In comparison, Table A contains results of testing the main cephalosporin antibiotics for injection (cephalotine, cephaloridine and cefazoline, 87/1, 87/4 and 541/296, an important example of a known compound with a carbamoyloxymethyl group at position 3 (87 / 359), two comparative compounds of formula IX, i.e. known from DK No. 2373/72 (586/1 and 640/1) of which the latter is the closest analogue of said application to the most preferred among the present compounds of formula I, namely cefuroxime (640/359) and finally the anti-isomer (750/359) similar to cefuroxime.

The code names for the tested compounds are shown in the following table:

Code Compound Note 87/1 (6R, 7R) -3-acetoxymethyl-7- (thiencephalotin 2-ylacetamido) ceph-3-em-4-carboxylic acid 87/4 (6R, 7R) -3-pyridinium methyl 7- (Thien-cephaloridin 2-ylacetamido) ceph-3-em-4-carboxylate 541/296 (6R, 7R) -3- (5-methyl-1,3,4-thiadiazole-cefazolin 2-ylthiomethyl) -7- (tetrazol-1-yl-acetamido) ceph-3-em-4-carboxylic acid 87/359 (6R, 7R) -3-carbamoyloxymethyl-7- (thien-2-ylacetamido) ceph-3-em-4 -carboxylic acid

Code Compound_ Note 640/359 (6R, 7R) -3-carbamoyloxymethyl-7-cefuroxime - Exs 1-3 [2- (fur-2-yl) -2-methoxyimino-acetate and 5-10 amido] ceph 3-em-4-carboxylic acid (syn isomer) 554/359 (6R, 7R) -3-carbamoyloxymethyl-7- Example (2-methoxyimino-2-phenyl-acetamido-ceph-3-em-4-carboxylic acid (syn-isomer) 576/359 (6R, 7R) -3-carbamoyloxymethyl-7- [2- Example 4 methoxyimino-2- (thien-2-yl) acetamido] -ceph-3-em-4 carboxylic acid (syn-isomer) 617/359 (6R, 7R) -3-carbamoyloxymethyl-7- (2- ex. 18-ethoxyimino-2-phenyl-acetamido) ceph-3-em-4-carboxylic acid (syn-isomer) ) 678/359 (6R, 7R) -3-carbamoyloxymethyl-7- (2- Ex. 16 phenoxyimino-2-phenyl-acetamido) ceph-3-em-4-carboxylic acid (synisorner) 695/359 (6R, 7R) -3-carbamoyloxymethyl-7- [2- Example 14 (fur-2-yl) -2-phenoxyimino-acetamido] τ ceph-3-em-4-carboxylic acid (syn-isomer) 586/359 ( 6R, 7R) -3-carbamoyloxymethyl-7- [2- Example 19 t-butoxyimino-2- (thien-2-yl) -acetamido] -ceph-3-em-4-carboxylic acid (syn isomer) ________ 750 / 359 (6R, 7R) -3-carbamoyloxymethyl-7- [2- anti-analog to cefuri (fur-2-yl) -2 -methoxyiminoacetamido] oxime (640/359) ceph-3-em-4-carboxylic acid (anti-isomer) 640/1 (6R, 7R) -3-acetoxymethyl-7- [2- (fur. 35) in 2- yl) -2-methoxyimino-acetamido] ceph-DK ans.2373 / 72 3- em-4-carboxylic acid (syn isomer) 586/1 (6R, 7R) -3-acetoxymethyl-7- [2- t-bux 14 in toxyimino-2- (thien-2-yl) acetamido] - DK ans.2373 / 72 ceph-3-em-4-carboxylic acid (syn-isomer) ______

The test organisms in Table A are selected taking into account the wide spread of cephalosporin antibiotic sensitivity. Thus, Staphylococcus aureus 663E is highly sensitive to all microbiologically active cephalosporins, while strains 1414E and 853E are penicillinase producing and generally more resistant and strain 1613E is methicillin resistant and not very sensitive to cephalosporins. Escherichia coli strains 573E, 851E, and 1507E are representative of those strains normally sensitive to cephalosporins, whereas strain 1193E has the transmissible factor of transmission which mediates a class III type of β-lactamase (M.H. Richmond and R.B.

Sykes, 1973 / Advances in Microbial Physiology / 31). Salmonella typhimurium 804E, Proteus mirabilis 431E and Haemophilus influenzae 1184E are sensitive strains representative of the respective species. Enterobacter cloacae 1321E and Klebsiella aerogenes 1522E are β-lactamase non-producing mutants of normally resistant / enzyme-producing strains. Serratia marcescens 1324E, Providence sp. 1497E and Haemophilus influenzae 1788E are ampicillin-resistant, β-lactamase-producing bacteria. All new cephalosporins must normally have an effect on the normal sensitive organisms; the greater the effect they have on the more difficult β-lactamase-producing bacteria, the greater their potential therapeutic value.

From Table A it can be deduced: a) Most of the compounds of formula I and the comparative compounds are active against the three normal strains of Staphylococcus aureus, but less against the methicillin resistant 1631E. Anti-compound 750/359 generally works poorly against staphylococci.

b) All the compounds except the anti-compound have good effect against the strains of Escherichia coli which do not form β-lactamase. Commercial Comparative Compounds and 87/359 exhibit high activity difference to Escherichia coli 1193E and 1507, respectively, having and lacking the R factor, while the compounds of Formulas I and IX have similar activity to these two strains, suggesting that they are resistant against the ΤΕΜ-β-lactamase, c) The commercial comparison compounds and 87/359 have little or no effect against Serratia, the β-lactamase-producing gram-negative strains used by Providence sp. and, in particular, Serratia marcescens, while compounds I and IX have good activity against them, confirming their β-lactamase stability. Also against the two test strains of Haemophilus influenzae, compounds I and IX are active in contrast to the commercial cephalosporins and 87/359.

d) The 1-oxides are generally not at par with the other compounds of formula I, but are valuable in comparison with known compounds against Proteus mirabilis, Providence sp. and Haemophilus.

Thus, in summary, all the tested compounds except the anti-compound have good effect against the test organisms that do not form β-lactamase, but only compounds I and IX against those that do.

Table B shows further results of in vitro testing of the compounds of the invention (formula I) and the commercial comparison compounds as well as 87/359.

It can be said about Table B: a) The strains of Staphylococcus aureus were penicillin sensitive and selected to reveal compounds with poor efficacy. All the compounds tested were highly active.

b) The β-lactamase producing strains of Klebsiella were resistant to 87/359 and the commercial cephalosporins. Cefuroxime had good activity, better than the other compounds of formula I.

c) The compounds of formula I generally had a better effect against Proteus (all β-lactamase producing) than the comparison compounds.

d) Cefuroxime was the most effective compound against the β-lactamase-producing strains of Escherichia coli, although the differences are not very large at the organism concentrations of 5 10 ml. If the bacterial concentration is increased to 100-fold, the MIC of the commercial cephalosporins and 87/359 increases significantly, but is essentially unaffected for the compounds I.

e) The test organisms of Salmonella and Shigella are β-lactamase resistant due to the R factors they carry. The same situation occurs with the Salmonella strain as with the strains of E. coli discussed above; The Shigella strains are more sensitive to the compounds I than the comparison compounds, even at the dose of 10 organisms per ml. ml.

f) The Citrobacter and Enterobacter strains tend to be very resistant to the comparator compounds, while the compounds I have good activity against these organisms.

g) The 1-oxides are particularly effective in relation to the comparative compounds against species of Proteus and Enterobacter.

In summary, Tables A and B show that the compounds of formula I and in particular cefuroxime (640/359) prepared by the process of the invention have far wider range of activity than the comparison compounds, which is mainly due to their very high β-lactamase stability.

147683 8 3 æ H) Cll U «Η WH ω o O PJH (DHHPP) B 1 + ¾ {DOHO (D t + HO _ _ ii g <; ϋ ri- tr ro 3 = SS5? U goh &> ro bho ( D tf p H · s ^ aCbr + CH-OO H · H · * tf ro hb ro tr ro h · „g tf H * 3 P> h $ n HO PJ 3

ID H O 3 Η Ω H tf C (D

^ C (II S H · pi ft I »H · .5

tf tn pi ti {D (D = - = fD

(D b ti P> P> ti c + P

w Η- Ό ο σ ffl * <Q m n- 3. (DF · li O tø = = = o (D Η) B Η OH tf I-1 3 Η Ω F · iQ O F- F- n- = 3 ® in in p 3 ro 3 3 O 3 ~ ~ 3 B (D p) H Hj H3 NB (DF · SO P> gs, s ro p ^ _ + I—> I—> I — il—> i— 1 Ϊ— * HH Η H 2 sJHiP'UlfcUlUCOOOUlUlHOliiOOm ti COCOt £> tOWtOtUOUl ~ J © lO <f'HUlH · 1-3 CO ^^ JJ ^ HtOH ^ HCO ^ UIWiUWOJ p ΜΗΜΜΜΜΜΜΗΜΗΗΜΜΜΜ σ - --- ~ = ΓΤΓ1- = - = - Π)

V OO | _I

| —I Cl tO ΗΛΛΛ ~ J J * j WOlWiwNit-WWi'it'O'OOOH \ fD> ° - - - H 3 - Η Η H p, --—---- zr £ s

to oo (D H

u u Ul U) AAAO O

^ .1_ιΗΟι ^. * »Λ. ^ 10Λ '· 6> · ΗΟΟΟΗ 1 \ htl - --- iCk o __Η Η H__Η -p

........ "V" "tf (Q

to to Ol p- \ | —i U1 Λ Λ © Λ »3 g! ^ <3iCOOtOHHOOHH> fc» OOOOOC ^ H pj I- ·

- - * - »\ (D

Ul U1 Η H fo H

-------- 7 ------ “B

H to ω oo ro ω to pi ηλλλ ui -j ti J ^ HUlO HfOtOtOi & iJ ^ if ^ UlOOOtO VD \ ^ ^

_ _ Η Η H

J_1 | __I HH H W «O l" h ίΡ 2wZ3u) aitotoui <Titoto touiui 03 OHOOHtouiuiHtouiUfOstoOoui iso g, <j_ __- _—— --—— I- Λ λ Η Λ Η ΟΙ

OOOOl ^ tOOOifc-tO ^^^ OOO ^^! ^ Ί m S

. "--- ΟωΦ0 wm ___ Η 01 01> ωρ, σ u> hhh η η tn \ ι + · toHi ^ HOiOiOlOOOOOlOOOOHHHiC» 00 "Ο m ΓΟ πι \ * 3 U7683 9 S æ * t> w» ti;> $ M co td cno

O 0 H (D Η H 3 0 w <+ H

(DOhOfDftH O O ua II 3 <hctO- (D3s = = ^ = = = V o O Η O Φ to H O (D tra H s Ό ft rt C Η · 0 3 H · W-

* · - 3 * ® h w <d o * <d η · „S

Λ * Η · 3 O Η O Η O 03 (D Η Ω 3 Η O H1 3 * ø Φ P <D 3 HO {+ O H- ll cr οι oi-i ro o = = = cd (D to Η O 0 H <+ 0 w η Ό oo * ro »<ow

rt »• (DP-HO '^ sssO

Φ Hi to Η O H 3 * H

3 HOF · iP O Η · F- (+ sp (DM) DO 3 (D 3 3 OP ~ · ~ · 3 to) DOK 1-3 1-3 N to (DF · td td OC gg (D 3 ^ + H Η Η H HH HH H Hg

sjH.c.u (> uiu <] oaiuiuiHoiit> a! ii H

ΟΟΟΟίχιΙΟΙΟΙΟΜΟΙπΌΟιΟσιΗυΐΗ · COifc.O.fe.HtOHit »HlOOlO (jOiC» lOU> tdtdtdtdtdtdbdtdtdtdtdtdtdtdtdtd 1-3

—-------: ---------- .— ~ ------. "·": ...... = 'Γ ----- O

λ λ ιο ιο σι θ '

ΟΟΟΗΗΜ> Ρ.> & · Ι0ιΡ.> Ρ> .ιΡ> ΟΟΟ00 ΙΠ ιΡ »(D

^^ kO 0 i-1

w w oi to υι in X

____> Λ Λ LO LO Λ Λ Λ LO ln Hi

OOHHtOHHCOtOCOOO OOOOOiP * lnln O

»- - - - 10 ιΡ- H

Ol Ul H H H \ et two

- - -------------- Oh

lo H lo Η H HHH H LO ln Γ + ΗΗΐοΗσιΗσισιοοσισισιοοοσιΐπ-ο - - - - »ιο σι οι οι w \ 2

H

--o AALOH HHH ΙΟ σι ,? -

οοιοΗΟοσιοοοοοοσισισισοοοοιπΗ H

- - - ιο —j Q, τ:

W ln Μ Μ M \ ° lQ

--3 g

Λ LO LO Η Η ΗΗΗΛ Λ LO σι g1 H

ΟΗΜΗΟοΗσισιοοσισισίοοοα ^ οπΌ> - - - LD 00 rr

Ln H Ol H \ $

__H

LO HHH LO A LO σι a ΗΗιΡ »Ηθοσισισιθοσισισιοοοοοίπνο 3 - 10 in *“ Η ΙΟ H \ F1 LOHLOHH HHH H LO ΙΠ !?

ΗΗΜΗσιΗσισιοοσισισιοοοσιιποο H

lo σι 3 ΙΠ Ln U1 \ g

___________ H

σι pj h

v> P> Hl W H

to Η O I

ΙΠ H tO \ o

tOHtOOOtOtOtOOtOtOCTlitiOOOOLnlO X

'in FJ

Ln Ln vo Qi 1

v σι o H

two to two «> Hi CO

LO Ln Ln HH HHH UlOI

HOOOit »CO (JiCJlOO (TiCricritOJ5-tOO \ 0

'LO X

to in K

l £> p 147683 10

Table B, MIC, µg / ml

Conn. of formula I

Known Compounds____

Organism no. 87/1 87/4 541/87/640/554 / __________ 296 359 _ 359 359_ S. aureus' 3972 <0.1 <0.1 <0.1 <0.1 <0.2 0.2 <0.1 748 < 0.1 <0.1 <0.1 <0.1 <0.1 <0 »1 864 <0.1 <0.1 <0.1 <0.1 <0.1 <0/1 K. Aerogenes 2972 125 62 31 62 8 31 "47876 125 62 31 31 8 16" 47877 250 250 62 62 8 16

Klebsiella sp. 1260E 250 62 62 250 4 8 P. Vulgaris Poole> 250 250 62> 250 16 16 "45343 31 62 16 62 4 4" 45780> 250 250 62> 250 31 31 P. Morganii 1376E> 250 250 62> 250 16 16 P ftiirabilis 1315E 16 31 4 8 4 8 P. Straightener 1484E 4 2 <0.1 16 <0.1 0.2 "1353E 31 8 4 125 <0.1 0.2 E. coli J53 16 16 4 8 2 4 R825 "SM239 884124" SM209 31 31 4 82 4

Salmonella sp. 1347E 2 2 0.5 2 4 8

Shigella flexneri 209A 250 125 31> 250 4 4

Shigella dysenteriae 1667E 250 62 250 250 31 31

Citrobacter freundii 159 250 125 125 250 2 8 "" 1506E 125 125 125.> 250 8 8 "" 367> 250 250> 250> 250 62 31

Enterobacter hafniae 1333E> 250> 250 125> 250 4 2 "6372 62 62 62 31 8 16

Enterobacter cloacae 1686E 16 16 2 62 8 31 "" 1015E 250 250 250> 250 8 16 "1683E> 250> 250 250> 250 8 31 147683 11

Table B continued, MIC, µg / ml__

Compounds of Formula I

ls ~ oxide oxide of 640/640 /

Organism no. 576/617/678/695/586/359 359 ___359 359 359 359 359___ S. aureus 3972 <0.1 <0.1 <0.1 <0.1 0.5 4 4 "748 <0.1 <0 , 1 <0.1 <0.1 <0.1 2 2 "864 <0.1 <0.1 <0.1 <0.1 0.2 2 1 K. Aerogenes 2972 31 62 62 62 31 125 62 "47876 31 31 62 62 125 125 31

„47877 31 62 62 62 62 ND ND

Klebsiella sp. 1260E 8 8 8 8 8 16 62 P. Vulgaris Poole 31 16 16 16 62 4 250 "45343 16 16 16 31 62 62 16" 45780 125 62 16 62 125> 250> 250 vo P. Morganii 1376E 31 16 31 16 16 4 250 P. rttirabilis 1315E 16 16 31 4 16 4 31 P. Straightening 1484E 0.5 0.5 1 1 1 <0.1 <0/1 "1353E 0.5 1 2 2 2 0.2 0.2 E. coli J53 8 8 16 8 16 16 16 R825 "SM239 8 16 16 8 16 2 16" SM209 8 4 8 4 4 4 31

Salmonella sp. 1347E 8 8 8 8 8 4 16

Shigella flexneri 209A 8 8 8 4 4 16 8

Shigella dysenteriae 1667E 31 31 16 16 16 31 62

Citrobacter freundii 159 8 8 16 8 8 62 125 "" 1506E 16 8 16 8 8 31 62 "" 367 62 16 31 31 31> 250 250

Enterobacter hafniae 1333E 4 4 8 8 8> 250 125 "" 6372 16 1 8 8 8 16 31

Enterobacter cloacae 1686E31 31 31 31 31 8 16 "" 1015E 16 8 16 16 16 16 62 "" 1683E 31 16 31 16 16 31 62 147683 12

One area where cefuroxime (640/359) seems particularly promising is its activity against strains of Haemophilus influenzae, and Table C shows a detailed overview of the action of cefuroxime against strains of this microorganism in comparison with commercial β-lacquer antibiotics. Since ampicillin has a good effect against strains of H. influenzae, a commercial sample of this antibiotic was used instead of cephaloridine. It is seen from Table C that cefuroxime is always more active than the comparison compounds.

Table C

----—---——- H. influenzae __MIC, ua / ml __ strain 640/359. AMP 87/1 541/296 1184 E 2 0.5 4 16 LH 141 0.5 0.25 0.25 8 LH 147 0.5 0.25 1 8 LH 150 0.5 0.25 2 8 LH 151 0 , 5 0.25 2 8 SM 137 0.5 0.25 2 8 LH 143 0.5 <0.125 0.25 8 LH 144 0.5 <0.125 0.5 8 LH 145 0.5 <0.125 0.5 8 LH 146 0.5 <0.125 2 8 LH 148 0.5 <0.125 0.5 8 LH 152 0.5 <0.125 1 8

248 6X 2 250 31 ND

4347X 2 250 31 ND

12214X 1 250 31 ND

12256 * 2 250 31 ND

23141X 2 250 31 ND

23142X 0.5 250 16 ND

23143X 2 250 31 ND

23144 0.5 62 31 ND

23145X 2> 250 31 ND

23366X 2 250 16 ND

23367X 0.5 250 16 ND

23368X 0.5 250 62 ND

23369X 0.5 250 31 ND

ChistyX 0.5 250 31 ND

| Birkert Q'5__250__31_i ^ 147683 13 x β-Lactamase Formers AMP = ampicillin ND - not determined

Cefazoline was not tested against these β-lactamase-forming strains because it had a poor effect on the sensitive strains and is sensitive to the β-lactamase that the ampicillin-resistant strains of H. influenzae form.

A further important site for cefuroxime has been tested against strains of Neisseria. Table D provides an overview of a number of representative strains and also provides results for the three commercial comparison compounds. It is clearly seen that cefuroxime always has high activity against strains of Neisseria, generally at a much higher level than the comparison compounds.

Table D

Art strain _MIC, yg / ml _ 640/359 87/1 87/4 [541/296

Neisseria meningitidis 891E 0.02 0.2 3.1 0.3

Neisseria catarrhalis 668E 0.2 0.1 0.8 0.1

Neisseria gonorrhoeae 35 0.01 0.16 2.5 1.25 * 157 0.005 0.31 2.5 "0.6

„27 0.005 0.31 ND ND

"33 0.02 ND 2.5 0.6" 28 0.02 ND 2.5 0.3 "13 0.02 ND 2.5 0.6" 48 0.02 ND 0.62 0.08 "1130 0.08 ND 2.5 1.25 "110 0.01 0.62 0.62 1.25" 163 0.01 0.62 0.62 1.25 "19 0.08 ND 2.5 1.25 "125 0.04 ND 2.5 1.25" 40 0.04 ND 2.5 1.25 "15 0.02 0.5 2.5 0.6 1 0.001 0.5 0.8 0.1" 2 0.002 0.5 0.8 0.1 »__3__0.002 0.5 __ <Μ ___ 0.05 j

Tables E and P show effect on MIC of cefuroxime and commercial comparison compounds of increasing the rate of infection with the test organism from 10 to 10 organisms / ml. It can be seen from this that in most cases the MIC for cefuroxime is only slightly altered using a 100 times the amount of microorganisms in the assay, whereas the comparator compounds generally have significantly lower activity against these high seed concentrations.

The comparative compounds of formula IX, i.e. representing DK-PA 2373/72, are not included in Table BF because these compounds will generally exhibit somewhat similar activity in vitro to the compounds I prepared by the process of the invention, having similar stability to β-lactamase. (a function of the nature of the 7 β-acylamido group).

However, the present compounds have a significant advantage over the compounds of formula IX, namely the advantageous property of having good in vivo stability, especially against esterases. This is particularly advantageous because a number of known cephalosporin compounds have the disadvantage of undergoing degradation in vivo and thereby deactivated before they can exhibit their full antibiotic effect. Table G shows the degree of stability of three representative compounds of the present Formula I and of the two Comparative Compounds of Formula X against a rat liver homogenate, a practical source of animal esterase. It is immediately apparent from the table that the three compounds of formula I are stable in the presence of such an esterase, whereas the comparative compounds are degraded to a considerable extent until ca. 50% in 4 hours.

The test was performed by incubating the test compound at a concentration of 100 µg / ml at 37 ° C in the presence of 2% rat liver homogenate. Control samples were incubated in the presence of physiological saline. Samples were taken from the test and control material after 0, 1, 2 and 4 hours and determined for antibiotic activity. The results are given as the percentage of the compound remaining after the specified time, calculated on the basis of control = 100%.

147683 15 s? in? »R> * s .H« Γ ϊ ΙΤΐΤΥϊΤΐΤΓ ^ Υ g

/? ffi ri · D * H -J Ui g> CT

Ω Ω P tf Ω O (5 i ^ toH ^ wMtoto § ®

3 g H H ® »O O HH-JWW ^ O iH

O H 0 O ri Η Η H M ^ hrt ri ri fi fi O φ Η- P- ril ra

i £ t P Ω O iQ H

tu tr p p> π> h. __———, is H · Φ CD O pj <s.

Η · Η Φ K 'ri H * m W | § XOOOOOOHOH 5 "e} 3 ^ ^ ^ - o p.

p · H Ul Ul to to Ul Ul Ul 2 ------ ri to Ui Ui <ΰ 4a.tO'X3td * .Ji.HCn 0) VQU1 O} nj to ^ aa-vioows t +.,; r hi lomsjoi ^ woto SS. v «OD σι to 03 Μ O 3H1 prx η o ω ω ud 3η_ ~ x (D Ό Ui & O OOOOOOtOOHUD ^

- ♦ I ------ pj ^ ^% "* O S

j ~ * (DW Ui Ln (O to Ui (ji'vl p ODHOOtOOOOO ^ tOO 3ui U1U1> ΰ UI g φ σι Φ _ Ό rfi. & tu »o Φ v OOHtOiUit»> &> OOH β W Ό - - O £ U, p ° Ul Ul Ul Η H «5 ~ m §

to OJ H o g fi X

υιΝοοοιυοοσιΟΟΦ. ^! pj p ^

U tD

Q i

3 V V V V V H

O toto to to to H P, • - ........................... = = === p, VUlUlVVUlUlUlO s 3.

rf ^ OOd ^ it ^ OOO Ό H S [v '• Q O s

two H ι-i -S

Ul Η σι H W O g τ-Φ ο ^ οο ^ σιίοσιΐσ ui g -—--- h P H | -i oo fi> 8 -s] g lQ ΛΛ ΛΛΛ AH 3 Pj

P OOOOOOOOO H G µ Π ft -------- Ul U

·, Η- HHHHHHHH 00 g

VV V V tOOfU

NDtOHH tOtOtOH ~ G ® U1 9 u-iuitorocnuiuiuioiQ M X1 OOU1U1MOOO —J \ ° 3 ffi. 52nd

Hi r +

ri- Λ Η P

fD OOOOOHHHO 3 --- ri - '. «. ·. ·. -jav P-HtOOltOtO g tO H to H 2 01 utoui w σι σι on ----—-—-— ~ - --- ffi OUlOit * tOif »tOtOUl 'ΛΛ Λ Λ Λ Λ Λ Ηι ¢ 0 ΟΟΟΟΟΟΟΟΜ 1 * J rf p.

Οι ^ *> ΙΟι ^ *. Η * Η Q

- ο G

-—--- to ~ J m Η H cn H o fi σιοοιοΝίοοσιοοοο υι --- c

Out of m ΛΛΛ ΛΗ ρ Ι_ι οοοοοοοοο ω Ν ·. (jn · two HHtOHHHtOH Ul Vø Ul ^

v σι H

tO (O to H> · ΗΗυισισυιυισο 10 cnoototoooto · -] J®

---- OOHOOtOtOtOO

** ^ «* · * H Ui cn Ul 147683 16

Table G

Stability to rat liver homogenate.

For example, _% left after_ ____0_t__l_t__2_t__4 t 586/1 96 65 60 45 640/1 92 59 54 47 640/359 100 100 100 100 695/359 100 100 100 100 554 / 359__100__3 £ 0__100__100

The compounds of formula X, i. those known from DK-PA 381/72 differ in two essential respects from the present. First, they are hydroxyoxymers or acyloxyoxymers, i.e. esters, while the present invention relates to oximeters. Second, the present compounds have a carbamoyloxymethyl group at position 3, while such group is not shown in the older application.

As is well known, the chemical properties of ethers and esters are quite different from the chemical properties of esters and alcohols in many respects and it is therefore not obvious to assume that one can add from the properties of esters to those of ethers, in the present case so much the less the 3-position is otherwise occupied.

It also turns out that there are biological differences between the known esters and the present ethers. The latter are more stable to esterase than the known compounds and have decreased serum binding.

Table H shows the stability to rat liver homogenate, determined in the same manner as described in Table G. It is seen as before that the present compounds are not attacked by the esterase while the known compounds do.

Table I shows the binding of the compounds to serum. The present compounds bind to a much lower degree to serum than those known in formula X. The practical utility of a compound as an antibiotic depends not only on its antibiotic activity, but also on its resistance to, among other things. esterases and lactamases, and of being available in the organism, an availability that is diminished when it is tightly bound to serum. Therefore, it is seen that the present compounds of formula I are advantageous over those of formula X

17 147683 in this regard.

The anti-compounds known from DK-PA 382/72 make the present compounds more distant than those already discussed. The compounds I prepared by the process of the invention have substantially higher antibiotic activity than the known anti-compounds as shown in Table J. In this, the listed anti-compounds from ans. 382/72 those which are structurally closest to the present compounds. It should be noted that the results given in Table J do not derive from parallel experiments, but are compiled from different series of experiments. However, they clearly show that the known anti-compounds have a significantly poorer effect, especially against gram-negative bacteria, than the present ones.

Table H

RA-C CONH__

I-o- * B J— «i rC

COOH

RA RB R ^% compound remaining after _0_1_2_4 t O -H -CH2OCOCH3 known 80 70 60 50 ^ -COCH3 -CH2OCOCH3 known 80 75 60 50 ^ -CH3 -CH2OCONH2 new 100 100 100 100 “CH3 -CH2OCONH2 new 100 100 100 100 ^ -ch2oconh2 new 100 100 100 100 147683 18

Table I

ABC

RRR% serum binding -H -CH2OCOCH3 known 90 -COC2H5 -CH2OCOCH3 known 80 "H -CH2OCOCH3 known 98 -COCH3 -CH2OCOCH3 known 95 -H -CH2OCOCH3 known 92 ^ Υ -CH3 -CH2OCONH2 new 49 O -C2H5 -CH2OCONH2 new 38 "0 -CH2CONH2 51 ^ ~ y ~ -CH3 -CH2OCONH2 new 30 19 147683 DK-ans. 382/72 ^ -p

N H

-— ------ Η O. _ I

-j σι oi to tv ^ 2 = 0

in I

• Oh

- § o σι σι σι σι σι σι σι td Β w ω ιο Η οι w w Ο Q. ι σι σι ω ο Ηι u> Η Ν \ ρ, Η Η Η Η Μ Η Η · '1- m Ρ

Di Ζ ω Η οο ® _ Μ (D Ζ— ~ λ ^ \ λ <λ ο π ο ί π ι Λ_ / “Μ ^ -ο Α * ή Υ * Γ» - · Μ ^ Β Τ · Νο - ο _____ ο to a β β β β a wi te to CO w

(D

H

H

ro H £

Dl ---—--- tr V H d OT ® tontooiotototn · i-1 »» ^ P> _ ui co σι σι d '-i

o H 4! * ro I

V H d OT

JOH Η C7l O H H 01 ·> »^ X N P) σι to co to σι d σι οι οι σι ti w (D · I>

---------- H

v v σι H _ i two to two --J, 2 = 0

oj f— 'σι σι ω σι ui oj o / I

toto o o Η o H 0 _ O O

h g b o

^ _____ M I

v V V V 1-3 M

to co co to W · O

σι σι σι σι σι σι 3 0 5 ο to ο ο to ο 40 40 γ1 \\ Ο Η s 1ΰ

μ. 3 (D

__ I-1 Μ ν οο 3 w w μη tO tO tO Od · * 5 tot- · σι σι w οι w H t + _ Ό

σι σι o o Η o h Η -> <.2— (D

is' / & H- O -i \ OT fl> ____! _ 8 Λ / ov in * tf bb> - 'rø toto H UH ·· / Hl σι η σι σι H to σι η Η 3' Η · to σι oo σι σι to μ- μ- ogtn Η B θ '

pi co (D

I O H

_----- O 01 v toifl BO® 2 to w pj · 2 ui i-3 σι σι w οι p 3 Β

to O OiUHif »H * 0 tO

H- ti 1! "" "" ""

1-1 HID B

W to Η H HS · to co σι σι oo σι oo n η-

Λ · d 0 (D Hi H

d

_____________ I

207 147683

The present invention

_— --— - “M

I — 1 i — 1 l — 1 Ϊ — 1 Ϊ — 1 I — 1 fT

two æ> - ~ J <r \ tn υισ \ σ> σιυισιυισι ^ ooHoo-'Jtnio ^ JiJ ^ y cn <! to oo £> tn σι o 2, \\\\\ >>> £ oj to ui. two to two to two years tntntntntntntnlntn »LDtOLDVDtOVOtOVO jg1 I- ·

Dl (0 0 g o g g o o o7 1 I I i O I O O ^ * o o JL> 4 ^ tt tt, tt otti I £

ffi tn I I H

^ ® w 11 Λ A M ^^

H o O O O O O ° WQJ £ L

*>> "- * ^ _ j: er ro to h to lh h in 25 <&

tn u, m SS H

1 C4 Λ Λ Λ ^ Hl

H O O O O O O O «O

two to M H Ol H to 2 S d * ^ ^ ss s __L_ £ '"~~ tn tt Η« H «8 σιοοοοσιοοιοσι * ^ r, 2

μ. H

____o h3 tt Η · p

2 0 ΤΗ Η Η Η H * 40 iQ

σισισισιοοοοσιοο ™ \ __ £ _ & oo 3 w w o C · | _j> 6 * H ft CTiCOOOOlCOOOOOifc * - 3 tt

H

_IN

'"*>. tr1 tt w H · · H H H 3 σ» oo co oo> t> oo 1-1 m'h

P

tOiQ tt. W p ·

A ιη Μ H

Η o O tO O O O O p. O

· »> '' '' Μ. H

totn to tn h to i tn tn tn -—- "Η Φ ΠΓ M 3 · ΛΛ ΛΛΛ 00ΝΗ * Η O O Μ O O O H Æ. p 3 '' '"' (D hh tn tn to tn in P «tn c 147683 21

Salts that can be formed from compounds of general formula I include inorganic base salts such as alkali metal salts, e.g., sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and organic base salts, e.g. The salts may also be resinates formed with, for example, a polystyrene resin or cross-linked polystyrene-divinyl-benzene copolymer resin containing amino groups or quaternary amino groups.

When R * in general formula I is a furyl group, it can be a fur-2-yl group or fur-3-yl group and when it is a thienyl group it can be a thien-2-yl group or thien-3-yl group.

Preferably, the group R1 is a fur-2-yl group.

2

As mentioned, the group R of formula I is an alkyl group of 1-4 carbon atoms, for example methyl, ethyl or t-butyl; a cycloalkyl group having 3-7 carbon atoms such as a cyclopentyl group; or a phenyl group.

The properties of the compounds of the invention make them useful for the treatment of various diseases caused by pathogenic bacteria in humans and animals. A particularly important compound is, as seen in Tables A and B, cefuroxime (640/359) or the syn isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2 - methoxyiminoacetamido] -ceph-3-em-4-carboxylic acid of the formula

Η H

O - " '00" - II

V J-1 J — CHpO.CO.NHp

Ndch3 cr

COOH

and suitably taking the form of an alkali metal salt, especially the sodium salt. The compound is resistant to the action of mammalian esterases and is therefore stable in human and animal bodies, as evidenced by high degrees of recovery of unchanged compound in the urine. In addition, the compound provides high serum concentrations after parenteral administration, both in humans and animals, and exhibits low serum binding. According to the invention, therefore, it is particularly convenient to prepare this compound, optionally in the form of a non-toxic salt, biologically acceptable ester, 1-oxide or solvate thereof.

Use of easily soluble base salts (e.g., alkali metal salts such as the sodium salt) of the compound is advantageous in therapeutic applications because of their rapid distribution in the body after administration by injection. According to the invention, therefore, the Na salt of ce furoxim is particularly conveniently recovered.

It has been found that the sodium salt of cefuroxime occurs in a number of different crystalline forms, including solvates, the preparation of which are all encompassed by the present invention.

The sodium salt is most conveniently prepared by bringing a solution of compound II into a polar organic solvent, e.g., dimethylacetamide, a mixture of such solvents, e.g., dimethylacetamide / acetone or dimethylformamide / methylalcohol-denatured ethanol, or an aqueous polar organic solvent, contact with a small molar excess of sodium 2-ethyl hexanoate dissolved in a suitable organic solvent (e.g., an alkanol such as ethanol, a ketone such as acetone or a chlorinated hydrocarbon such as methylene chloride, an ester such as ethyl acetate or an ether such as dioxane), conveniently at room temperature , after which the precipitated salt is collected, possibly after cooling the solution, for example to 4 ° C.

Where substantially anhydrous solvents are used in this process, Form I syn isomers of sodium (6R, 7R) -3-carbamoyloxymethyl-7- / 2- (fur-2-yl) -2-methoxyiminoacetamido-7 ceph-3-em-4-carboxylate, and this material contains approx. 1.5% water. If the solvent system contains more than approx. However, 2% water is obtained from Form II salt, which usually contains approx. 2% water. If the solvent system contains more than approx. 60% dioxane is usually obtained from Form III salt, and this material comprises a dioxane solvate containing ca. 1 mole of dioxane, although it is possible that salt of Form II may be formed if a water-wet solvent system is used at elevated temperature (e.g. 60-80 ° C). Crystallization of amorphous freeze-dried sodium (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carb oxylate (syn -isomer) from suitably dry, aqueous or dioxane-rich solvent systems similarly yields salt of Form I, Form II or Form III, respectively.

Exposing the salt of Form I to water vapor (e.g., at a relative humidity of 75%) causes the salt to absorb additional water and undergoes a change in crystal shape which generally leads to a salt of Form IV. The resulting material contains approx. 4% water (i.e., about 1 mole) and is believed to be a hydrate. This change is reversible so that the salt of Form IV can be converted to salt of Form I for example by drying in a vacuum over a desiccant such as phosphorus pentoxide. Salt of Form II does not absorb additional water when exposed to steam, but can be converted to salt of Form I by heating (e.g., for about five minutes) a suspension of Form II material in near-boiling methanol.

Form III salt obtained by reacting compound II and sodium 2-ethylhexanoate in dioxane-rich solvent systems as described above is usually precipitated as a gel which can be dried in vacuo to give a very low bulk weight solid with little or no crystallinity. However, the crystalline salt of Form III can be obtained by treating an aqueous solution of the sodium salt with a substantial excess (e.g., about 3 volumes) of dioxane, optionally with a small amount of ethanol, appropriately collecting the resulting white needle-shaped crystals. after cooling to a reduced temperature (e.g., 4 ° C) and washing the product with dioxane and then drying the crystals, e.g., in vacuo at 20 ° C.

The salt of Form III is hygroscopic and loses by exposure to water vapor (e.g. at a relative humidity of 75%) all dioxane present and forms materials of Form IV which can then be dried (e.g. over phosphorus pentoxide) to give salt of Form I. crystalline material of Form III is treated in this way, the crystal behavior of the product appears to be maintained during the conversion process. Salt of Form III can also convert salt of Form I by heating a suspension of Form III material in near-boiling methanol; this conversion leads to loss of crystallinity if crystalline material of Form III is used.

24

The four forms of the syn-isomer of sodium (6R, 7K) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylate described above is characterized by the following X-ray powder patterns (d distances and intensities) and IR spectroscopic data: X-ray powder patterns

Camera - Debye-Scherrer, radius 114.6 mm Radiation - copper K ^ = 1.5418 k Intensity (I) for visual comparison with a calibrated standard

Form I

dl d I d I

8.33 80 3.82 24 2.40 10 7.44 4 3.62 28 2.35 10 6.85 45 3.47 28 2.26 4 6.38 5 3.32 10 2.20 3 5.86 4 3.19 10 2.11 8 (wide) 5.36 4 3.05 4 2.04 3 4.82 100 2.93 14 (wide) 1.94 4 4.56 35 2.72 8) poor 1.89 5 4.36 6 2, 69 10) dissolved 1.82 6 (wide) 4.19 40 2.57 9 1.77 2 3.95 26 2.47 6

Form II

dl 4 1 d I

8.78 60 4.20 10 2.77 6 7.81 9 3.76 20 (wide) 2.32 3 (wide) 6.65 25 3.49. 14 2.19 2 4.68 100 (wide) 3.07 6 (wide) 2.08 2 4.45 10 2.91 8 147683 25

Form III

dl dl d I

14.98 60 5.57 20 3.47 5 12.95 40 4.98 40 3.32 10 10.16 20 4.73 60 3.26 30 8.23 45 4.29 20 3.13 17 (wide ) 7.52 5 4.16 100 2.43 10 6.61 65 3.81 25 (wide) 2.15 15 6.08 3 3.60 20

Form IV

dl d I d I

8.85 70 4.65 100 2.93 4 7.80 6 4.30 25 2.76 12 7.15 25 4.01 25 2.62 1 6.01 20 3.75 35 2.41 2 5, 06 18 3.10 1 2.30 3 IR Spectra

Spectrometer - Perkin-Elmer 521, range 4000-650 cm 2

Spectra listed for nujol slurries, as bands attributable to nujol are deleted.

Form I

3520 w 1620 s 1328 s 1112 m 882 s 3460 mx 1590 s 1284 m 1076 m 878 sh 3370 m 1556 m 1262 m 1054 sh 838 w 3265 s 1534 s 1248 mx 1042 s 814 w 1770 sh 1480 m 1170 m 1004 m 790 w 1752 s 1410 s 1152 m 978 m 778 w 1706 s 1400 s 1134 w 918 w 754 m 1660 s 1338 s 26 167683

Form II

3526 m 1642 sh 1284 m 1080 m 882 m 3492 w 1624 s 1268 m 1058 m 878 wx 3364 m 1544 s 1240 mx 1045 m 840 w 3250 m 1478 m 1172 m 1005 m 818 w 1758 s 1412 s 1152 m 980 m 792 w 1695 s 1398 s 1142 m 954 w 752 m 1665 s 1332 s 1112 m 920 w

Form III

3465 m 1632 m 1285 m 1078 m 878 s 3415 m 1618 s 1260 mx 1058 s 836 w 3345 m 1552 m 1230 m 1048 s 820 w 3275 m 1532 s 1225 m 1014 m 800 w 3200 m 1482 m 1196 w 985 m 790 w 1780 s 1412 s 1180 m 938 w 768 m 1702 s 1395 s 1155 m 888 m 748 m 1660 s 1326 sx 1124 s

Form IV

3585 w 1620 s 1330 s 1078 m 882 m 3520 w 1594 s 1285 m 1058 m 878 w 3370 m 1555 m 1264 m 1042 m 838 wx 3260 s 1540 m 1240 m 1008 m 818 w 1758 s 1478 m 1172 w 980 m 788 w 1712 s 1410 s 1152 m 956 w 752 m 1664 s 1400 s 1114 w 920 w

Short explanation: s = strong, sh = shoulder, m = medium, w = weak.

x means bands characteristic of each of the crystal forms.

Particularly valuable is the salt of Form II of cefuroxime, and according to the invention, it is therefore convenient to proceed as set forth in claim 4.

In cases where insoluble salts of compound I are desired for a given application, for example, for use in depot preparations, such salts may be formed in a conventional manner, for example, with appropriate organic amines.

For example, when using the process (a) of claim 1, R 11 in the compound of formula III may be the residue of an ester-forming aliphatic or araliphatic alcohol or an ester-forming phenol, silanol or stannanol, or a symmetrical or mixed anhydride group derived from a suitable - = 12 the acid. When R is an N-protecting group, it may be, for example, an acyl group, in particular a lower alkanoyl group such as acetyl, a halogen substituted lower alkanoyl group such as mono-, di- or trichloroacetyl, or a chlorosulfonyl group. For example, an acid addition salt of compound III may be formed with a mineral acid such as hydrochloric, hydrobromic, sulfuric, nitric or phosphoric or an organic acid such as methanesulfonic or toluene-p-sulfonic acid.

Non-toxic salts of compounds of general formula I can be formed in any convenient manner, for example, by methods well known in the art. Thus, for example, base salts can be formed by reacting the cephalosporic acid with sodium or potassium 2-ethylhexanoate. Biologically acceptable ester derivatives can be formed by conventional esterification agents. 1-Oxides can be formed by treating the corresponding cephalosporin sulfide with a suitable oxidizing agent, for example, with a peracid such as metaperiodic acid, peracetic acid, monoperphthalic acid or n-chloroperbenzoic acid or with t-butyl hypochlorite suitably in the presence of a weak base such as pyridine.

Compounds of general formula I may conveniently be prepared by condensing a compound of formula III with an acylating agent in the form of an acid halide, in particular an acid chloride or an acid bromide corresponding to the acid IV. Such acylation can be carried out at a temperature between -50 and + 50 ° C, preferably at a temperature between -20 and + 30 ° C. The acylation can be carried out in aqueous or non-aqueous media.

Acylation with an acid halide can be carried out in the presence of an acid-binding agent (e.g., a tertiary amine such as triethylamine or dimethylaniline, an inorganic base such as calcium carbonate or sodium carbonate or an oxirane, preferably a lower 1,2-alkylene oxide such as ethylene oxide or propylene oxide). to bind hydrogen halide released by the acylation reaction.

147683 28

The free acid form of the compound of general formula IV can itself be used as an acylating agent. Such acylations are conveniently carried out in the presence of, for example, a carbodiimide such as N, Ν'-diethyl, dipropyl or diisopropylcarbodiimide, N a carbonyl compound such as carbonyl diimidazole; or an isoxazolinium salt such as N-ethyl-5-phenylisoxazolinium 3'-sulfonate or N-t-butyl methylisoxazolinium perchlorate. The condensation reaction is suitably re-introduced in an anhydrous reaction medium, for example methylene chloride, dimethylformamide or acetonitrile.

The acylation can also be carried out with other amide-forming derivatives of the free acid IV, for example, a symmetrical anhydride or a mixed anhydride, for example with pivalic acid or formed with a haloformate such as a lower alkyl haloformate. The mixed or symmetrical anhydrides can be developed in situ. Thus, for example, a mixed anhydride can be prepared using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. Mixed anhydrides can also be formed with phosphoric acids (e.g., phosphoric acid or phosphoric acid), sulfuric acid, or aliphatic or aromatic sulfonic acids such as p-toluenesulfonic acid.

When a starting material of the general formula V is used, suitable carbamoylating agents include isocyanates 13 13 of the general formula R .NCO (VIII), wherein R is a labile substituent group; such carbamoylating agents serve to form at the 3-position an N-protected carbamoyloxymethyl group of the formula -CH2O.CO.NHR where R has the meaning given above, which group can then be converted to the desired unsubstituted 3-carbamoyloxymethyl group by the following: 13 following the cleavage of the group R, for example by hydrolysis. Labile groups R13 that can be readily cleaved by such subsequent treatment are i.a. chlorosulfonyl and bromosulfonyl; aralkyl groups such as benzyl, p-methoxybenzyl and diphenylmethyl; t-butyl; halogenated lower alkanoyl groups such as dichloroacetyl and trichloroacetyl; and halogenated lower alkoxycarbonyl groups such as 2,2,2-trichloroethoxycarbonyl. Groups R of this type, with the exception of aralkyl groups such as diphenylmethyl, can generally be cleaved by acid-catalyzed or base-catalyzed hydrolysis, for example by means of sodium bicarbonate. Halogenated groups such as chlorosulfonyl, chloroacetyl and 2,2,2-trichloroethoxycarbonyl can also be reductively cleaved, while such groups as chloroacetyl can also be cleaved by treatment with thioamides such as thiourea. Aralkyl groups such as diphenylmethyl are conveniently cleaved by treatment with acid, for example a strong organic acid such as trifluoroacetic acid.

The carbamoylating agent of general formula VIII is conveniently used in excess, e.g., at least 1.1 mole relative to the compound of general formula V. The carbamoylation may be supported by the presence of base, e.g., a tertiary organic base such as a tri- (lower alkyl) amine , e.g., triethylamine, or using the acid V in the form of an alkali metal salt, e.g., the sodium salt, although such support may not be necessary in the case of more active isocyanates, e.g., compounds of general formula VIII

Wherein R is a highly electron withdrawing group such as chlorosulfonyl or trichloroacetyl. Thus, carbamoylations involving the reaction of a free acid V with excess of isocyanate VIII, wherein R is such a group as chlorosulfonyl or trichloroacetyl, are of particular practical advantage due to the simplicity of the reaction conditions, as there is no need for temporary blocking. and subsequent unblocking of the carboxy group at the 4-position of the cephalosporin and since the electron withdrawing group R of the resulting N-protective 3-carbamoyloxymethylcephalosporin product is readily removed by, for example, hydrolysis with aqueous sodium bicarbonate.

It should be noted that it may be appropriate to maintain or even introduce an N-substituting group R during conversions of intermediates produced 3-carbamoyloxymethyl compounds to minimize undesired side reactions involving the carbamoyloxymethyl group.

Another practically useful carbamoylating agent is cyanoic acid, which can conveniently be formed in situ from, for example, an alkali metal cyanate such as sodium cyanate, the reaction being facilitated by the presence of an acid, for example a strong organic acid such as trifluoroacetic acid. Cyanic acid corresponds in effect to the compound of 13. the general formula VIII wherein E represents hydrogen and therefore causes the conversion of compounds of the general formula VIII directly to their 3-carbamoyloxymethyl analogues.

3-Hydroxymethyl starting material for use in this embodiment of the process of the invention can be prepared, for example, by the methods described in British Patent No. 1,121,308 and Belgian Patent No. 783,449.

Any blocking group which is substituted on the 4-carboxy group in compounds of general formulas III, V or VI is suitably a group which can be readily decomposed at a later stage of a reaction sequence and is advantageously a group containing 1 -20 carbon atoms. Suitable blocked carboxyl groups are well known in the art of chemistry, and a list of representative groups is included in the aforementioned Belgian Patent Specification No. 783,449. Examples of preferred blocked carboxyl groups are aryl lower alkoxycarbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl; lower alkoxycarbonyl groups such as t-butoxycarbonyl; and lower haloalkoxycarbonyl groups such as 2,2,2-trichloroethoxycarbonyl. The carboxyl blocking group may later be removed by any of the appropriate methods described in the literature; thus, for example, acid-catalyzed or base-catalyzed hydrolysis is useful in many cases as well as enzymatically catalyzed hydrolyses.

Where at the end of a given preparation sequence a sulfoxide analogue is obtained for a compound of general formula I, conversion to the corresponding sulfide can be carried out, for example, by reduction of the corresponding acyloxy sulfonium salt or alkyloxy sulfonium salt prepared in situ, for example by reaction with acetyl chloride in the case of a acetoxysulfonium salt, the reduction being carried out, for example, by sodium dithionite or by iodide ions (e.g. as 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.

When the reaction product is a ceph-2-em-4-carboxylic acid ester, the desired ceph-3-em compound can be obtained by treating the former with a base.

The antibiotic-active compounds produced by the method of the invention can be reprocessed for administration in any convenient manner by analogy with other antibiotics.

For veterinary use, the compositions may, for example, be worked up as intramammary preparations on either a long-acting or rapidly delivering basis.

Generally, the compositions may contain from 0.1% of the active substance upwards, for example 0.1-99% and preferably 10-60% of the active material, depending on the method of administration. If the preparation is in dosage units, each unit preferably contains 50-1500 mg of the active substance. The dose used to treat adult humans will preferably be in the range of 500-4000 mg per day, depending on the route of administration and the frequency of administration.

The present compounds may be administered in combination with other compatible drugs such as antibiotics, for example, penicillins, tetracyclines or other cephalosporins.

147683 32

The following compounds are valuable as intermediates in the preparation of antibiotic-active compounds of general formula I: diphenylmethyl- (6R, 7R) -7-amino-3-trichloroacetylcarbamoyl-oxymethylceph-3-em-4-carboxylate and toluene-β the sulfonic acid salt thereof; diphenylmethyl (6R, 7R) -7-amino-3-carbamoyloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt; t-butyl (6R, 7R) -7- [2- ([uro-2-yl) -2-methoxyiminoacetamido] -3-hydroxymethylceph-3-em-4-carboxylate (syn isomer); (6R, 7R) -7-amino-3-kloracetylkarbamoyloxymetylceph-3-em-4-carboxylic acid; and (6R, 7R) -7-amino-3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid.

Some examples serve to elucidate the method of the invention. In the examples, melting points are determined on a Kofler block.

A) Preparation of starting materials

Starting material 1 a) Diphenylmethyl (6R, 7R) -7- (thien-2-ylacetamido) -3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylate 13.2 g (70 mmol) of trichloroacetyl isocyanate was added under stirring. suspension of 26.0 g (50 mmol) of diphenylmethyl (6R, 7R) -3-hydroxymethyl-7- (thien-2-ylacetamido) -ceph-3-em-4-carboxylate in 600 ml of acetone at 20 ° C. The solid dissolved soon and after stirring at 20 ° C for one hour it was cooled for one hour and the resulting solid was filtered off and washed with ether to give 33.1 g (93/0 of the title compound having a melting point of 183-184 ° C. o ^ 1 = + 24 ° (c = 0.95 in DMSO) λ infl (ethanol) 235 nm (i = 14,500) and λ infl (ethanol) 256 nm (δ 8.820).

IR, NMR and microanalytical data confirmed the structure as being the title compound.

B) Diphenylmethyl- (6R, 7R) -7-amino-3-trichloroacetylcarbamoyl-oxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt 31al (0.384 mol) anhydrous pyridine was added to a solution of 20 g (96 mmol) of phosphorus pentachloride in 300 ml of dry dichloromethane at 3 ° C. The suspension was stirred for 10 minutes at 3 ° C and then added 22.5 g (32 mmol) of diphenylmethyl- (6R, 7R) -7- (thien-2-ylacetamido) -3-trichloroacetylcarbamoyloxymethylceph-3-em-4- carboxylate; the reaction mixture was stirred at ca. 2 ° C for one hour. The dark solution was slowly poured into a cold (0 ° C) anhydrous mixture of 80 ml of methanol and 200 ml of dichloromethane keeping the temperature below 5 ° C. The temperature of the solution was then allowed to rise to 23 ° C and after stirring the solution at this temperature for one hour 200 ml of water was added. The organic layer was separated and washed with 2N sulfuric acid, water, sodium bicarbonate solution and water, dried over magnesium sulfate and evaporated in vacuo. The resulting oil was dissolved in ethyl acetate and a solution of 6.0 g (31.5 mmol) of toluene-p-sulfonic acid monohydrate in ethyl acetate was added. The combined solution (about 350 ml) was poured into ca. One liter of diethyl ether and the resulting solid were filtered off and dried in vacuo to give 17.2 g (72%) of the title compound, mp 150-153 ° C; α1 = + 7.5 ° (c = 0.82 in DMSO). λ max (ethanol 263 nm (£ = 7600) and K infl (ethanol) 267 nm (.t = 7350).

IR, NMR and microanalytical data confirmed that the structure of the product was the title compound.

Evaporation of the filtrate and trituration of the residue with ethanol gave 3.2 g (14.2%) of crude starting material.

c) Diphenylmethyl (6R, 7R) -7-amino-3-carbamoyloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt 17.2 g (22.7 mmol) of the toluene-p-sulfonic acid salt of the Nylmethyl (6R, 7k) -7-amino-3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylate was dissolved in a mixture of 900 ml of anhydrous methanol and 45 ml of acetyl chloride and left at 20 ° C for five hours. Removal of the solvent under reduced pressure gave an oil which was dissolved in dichloromethane. This solution was shaken with aqueous sodium bicarbonate solution and then washed with water. 4 * 3 g (22.7 mmol) of toluene-p-sulfonic acid monohydrate was added and the solvent was evaporated in vacuo. The residue was dissolved in ca. 150 ml of warm isopropanol and the solution was poured into ca. 600 ml of diisopropyl ether. The precipitated solid was filtered off and dried in vacuo to give 8.9 g (64%) of the title compound, mp 110-112 ° C; = -14 ° (c = 1.0 in CHCl3). Amax (ethanol) 259 nm (é = 6120) and λ infl. (ethanol) 227 nm (e = 15-800).

IR, NMR and microanalytical data confirmed that the structure of the product was the title compound.

Starting material 2

Diphenylmethyl (6R, 7R) -7-amino-3-carbamoyl oxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt

A stirred suspension of 156 g (0.75 mole) of phosphorus pentachloride in 1.5 liters of dry dichloromethane was cooled in an ice bath and treated with 60.5 ml (0.75 mole) of pyridine at such a rate that the temperature of the mixture remained at ca. 20-25 ° C. The mixture was stirred and cooled to 8 ° C and 354.5 g (0.5 mol) of diphenylmethyl- (6R, 7R) -7- (thien-2-yl) -acetamido-3-trichloroacetylcarbamoyloxymethyl was added portionwise over ten minutes. ceph-3-em-4-carboxylate. The mixture was stirred at ca. 8 ° C for 3/4 hour and then added over ten minutes to a stirred mixture of 225 ml (2.5 moles) of butane-1,3-diol and 500 ml of dichloromethane, pre-cooled to -20 ° C so that the temperature of the mixture is kept in the range between -15 and -20 ° C. The cooling bath was removed and the mixture was stirred at about - 10 ° C for twenty minutes. 1 liter of water was added and the biphasic mixture was stirred for 30 minutes. The aqueous phase was extracted with 2 x 500 ml of dichloromethane and the organic phases were washed successively with 1 liter of 2N hydrochloric acid, then combined and evaporated to a brown gum. The gum was dissolved in 3.6 liters of methanol and this solution was stirred and treated with 1.2 liters of saturated aqueous sodium hydrogen carbonate solution over a period of ten minutes. The mixture was stirred at ca. 20 ° C for 1/2 hour and a small amount of brown solid was removed by file. The yellow filtrate was concentrated in vacuo (bath temperature not above 40 ° C) to ca. 1.5 liters and 15 liters of water were added. The resulting suspension was cooled in the refrigerator for one hour and the yellow solid was filtered off, washed well with water, sucked as dry as possible and dried in vacuo at 40 ° C for 24 hours. The fatty solid thus obtained, followed by 81 g (0.425 mole) of toluene-p-sulfonic acid monohydrate, was added to 2 liters of stirred chloroform. After a few minutes, the toluene-β-sulfonic acid salt began to crystallize. Stirring was continued for another 30 minutes after which the water was azeotropically removed in vacuo with continuous replacement of the chloroform to maintain a volume of 2 liters. The suspension was refrigerated overnight and the product filtered off, slurry washed with 2 x 250 ml of chloroform, filtered again, washed by evaporation with 250 ml of chloroform and dried in vacuo at 40 ° C to give the title compound as a pale white crystalline solid in an amount of 237 »8 g (74.1%). kmax (ethanol) 262 nm (f = 7250). The NMR spectrum (Me 2 SO-d 6) indicated the presence of 0.25 mole of chloroform.

Starting material 3 (6R, 7R) -7-Amino-3-carbamoyloxymethylceph-3-em-4-carboxylic acid.

300.0 g (0.44 mole) of diphenylmethyl (6R, 7R) -7-amino-3-carbamoylloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt, solvated with ca. 0.6 mole of chloroform was added portionwise over 30 minutes to a stirring mixture of 300 ml of trifluoroacetic acid and 300 ml of anisole which was in a water bath at 20 ° C. The temperature of the mixture increased from 23 ° C to 28 ° C during the first 20 minutes but decreased to 26 ° C at the end of the addition. The golden yellow solution was stirred for one hour and the temperature dropped to 21 ° C, then added to a stirring mixture of 1.5 liters of ethyl acetate and 1.5 liters of water, placed in an ice bath. The pH of the stirred mixture was adjusted to 3.8 over ten minutes using ammonia water (density 0.880) and the temperature rose to 38 ° C. The suspension was stirred and cooled to 10 ° C over 1/4 hour, then filtered. The cream colored solid was washed with 750 ml of water and 4 x 200 ml of ethyl acetate and dried in vacuo to give 115.6 g (96.2%) of the title compound. -Vmax (pH 6 phosphate) 265 nm (λ = 7750). The purity was evaluated by 99.7% by high pressure liquid chromatography (HPLC).

Microanalytical data confirmed that the structure was the title compound.

Starting material 4 a) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-hydroxymethylceph-3-em-4-carboxylic acid

A stirred solution of 62.00 g (about 100 mmol) of ca. 67% pure (6R, 7R) -7- (R-5-amino-5-carboxypentanamido) -3-hydroxymethylceph-3-em-4-carboxylic acid mono-potassium salt in 300 ml of water was cooled to + 5 ° C and treated with a solution of 17.4 ml (150 mmol) of benzoyl chloride in 200 ml of acetone over 25 minutes. The pH of the reaction mixture was maintained at 8.2-8.5 by controlled addition of 30% w / v aqueous tricalcium orthophosphate. The mixture was stirred for a further ten minutes and then covered with 140 ml of ethyl acetate, and the pH was reduced to 5.6 with orthophosphoric acid. The layers were separated and the aqueous portion was washed with more ethyl acetate (2 x 400 ml). The aqueous portion was diluted with 2 liters of water, covered with 2 liters of ethyl acetate and the pH of the stirred mixture was brought to 2.0 with orthophosphoric acid. The layers were separated and the aqueous layer was extracted with an additional 3 x 1500 ml of ethyl acetate. The combined extracts were washed with 800 ml of saturated brine, dried and concentrated in vacuo to a volume of 300-400 ml. The resulting slurry was stirred with 2 liters of ether for 20 minutes and then filtered.

The collected solid was washed with ether (2 x 250 ml) and dried in vacuo (1 mm Hg) to give 54.95 g (88.6%) of the title compound as a white solid, aD = +74 ° (c = 1.00 in dioxane); Amax (pH 6 buffer) 231 nm (E ^ = 275), 266 nm (inflection, E ^ cm = 145) · The NMR spectrum (Me₂SO-dg) showed the presence of ca. 20% lactone impurity and ethyl acetate (about 0.4 mole).

B76 (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-chloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid monosodium salt 25.46 g of the product of (a) was treated with 9.00 g (75 mmol) of chloroacetyl isocyanate in 92 ml of dry acetone. The resulting solution was stirred for 25 minutes at ca. 20 ° C, then cooled to ca. 5 ° C over five minutes and treated with a solution of 8.47 g (51 mmol) of sodium 2-ethyl hexanoate in 51 ml of acetone. The crystalline suspension was stirred at ca. 5 ° C for five minutes and the solid was collected by filtration, washed with 80 ml of acetone and 250 ml of ether and then dried in vacuo (1 mm Hg) to give 27.23 g (107%) of the title compound ; = + 72.0 ° (c = 1.00 in 3% aqueous NaHCO 3); kmax (pH 6 buffer) 227 nm (E ^ e = 249), 261 nm (inflection, E ^ = 105).

The nuclear magnetic resonance spectrum (MegSO-dg) showed the presence of ca. 35% lactone impurity and approx. 1.0 mole of chloroacetamide.

C) (6R, 7R) -7-Amino-3-chloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid

A suspension of 24.77 g of the product of (b) in 320 ml of dry methylene chloride under nitrogen was cooled to ca. 10 ° C with stirring. 17.60 ml (218.0 mmol) of pyridine was added then 16.80 ml (139.2 mmol) of dichlorodimethylsilane and the light brown suspension was stirred at ca. 20 ° C for twenty minutes and then cooled to -17 ° C. Phosphorus pentachloride (10.84 g (52.0 mmol)) was added and the mixture was stirred at a temperature between -17 and -23 ° C for two hours. 6.48 ml (80.4 mmol) of pyridine were added and the mixture was added to methanol (104 ml + 20 ml wash liquids, pre-cooled to -35 ° C) at a rate such that the temperature of the stirred solution did not exceed -10 ° C. The stirred solution was allowed to rise to a temperature of + 2 ° C over 25 minutes and then 88 ml of water was added and the pH of the mixture was adjusted from 0.6 to 3.8 with ammonia water (density 0.880 ). The resulting biphasic mixture containing a precipitated solid was cooled for one hour and then filtered. The solid was washed successively with 100 ml of 50% aqueous methanol, 80 ml of methanol and 40 ml of methylene chloride and then dried in vacuo (1 mm Hg) to give 6.86 g (27.7%) of the title compound. as a cream powder, = + 48 ° (c = 1.04 in MeoSO); Λ max (pH 6 buffer) 237.5 nm (E + = 149), 261.5 nm (EI = = 145).

b) Examples

Example 1 a) Diylenylmethyl (6R, 7R) -3-carbamoyloxymethyl-7- [2- (iodo-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate (syn isomer) ) 147683 39

Method I

Crude diphenylmethyl (6R, 7R) -7-amino-3-carbamoyloxymethyl-ceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt, obtained from the corresponding 3-trichloroacetylcarbamoyloxymethyl compound (25.0 g, 0.33 mol) was dissolved in a mixture of ethyl acetate and aqueous sodium bicarbonate solution. The organic layer was separated, washed with water, dried over magnesium sulfate and evaporated on a rotary evaporator to give 11.5 g (0.262 mol, 77%) of diphenylmethyl- (6R, 7R) -7-amino-3-carbamoyloxymethylceph-3- em-4-carboxylate as a foam.

5.32 g (0.312 mol) of the syn isomer of 2- (fur-2-yl) -2-methoxyiminoacetic acid in 100 ml of dry dichloromethane were added to a solution of this amine in 50 ml of dichloromethane, cooled to 3 ° C. followed ten minutes later by a solution of 6.5 g (0.312 mole) of DL-dicyclohexylcarbodiimide in 30 ml of dichloromethane. The reaction mixture was stirred in an ice bath for 45 minutes during which time the solid was crystallized (presumably Ν, Ν'-dicyclohexylurea). This solid was filtered off and discarded, and the filtrate was washed with aqueous sodium bicarbonate solution and water, dried over magnesium sulfate and evaporated to dryness. The residue was triturated with ethanol to give 10.6 g of a crude product which was purified by chromatography on 1 kg of silica gel. Elution with 10% acetone in dichloromethane removed nonpolar impurities and fractions eluted with 20% acetone in dichloromethane gave 4.8 g (31%) of the title compound, mp 199-202 ° C; = + 14 ° (c = 1.0 in DMSO); Xmax (ethanol) 277 nm (£ = 18,600) and \ infl. (ethanol) 270 nm (e = 17,900).

IR, NMR and microanalytical data confirmed the structure as being the title compound.

Method II

1.86 g (18.4 mmol) of triethylamine was added to a solution in 35 ml of dichloromethane of 3.1 g (18.4 mmol) of the syn isomer of 2- (fur-2-yl) -2-methoxyiminoacetic acid. After cooling this solution in an ice bath for five minutes, 1.57 ml (18.4 mmol) of oxalyl chloride and a drop of N, N-dimethylformamide were added. After 1/2 hour, solvents were removed under reduced pressure and the solid residue dried for one hour in vacuo. 150 ml of anhydrous ether was added to dissolve the acid chloride that had been formed and the insoluble triethylamine hydrochloride (2.5 g) was filtered off. The ether was evaporated on a rotary evaporator and the oily residue redissolved in dichloromethane.

3.9 g (14.7 mmol) of diphenylmethyl (6R, 7K) -7-amino-3-carbamoylloxymethylceph-3-em-4-carboxylate-toluene-p-sulfonic acid salt was dissolved in anhydrous dichloromethane. This solution was shaken with aqueous sodium bicarbonate solution, washed with water and dried over magnesium sulfate. To this solution of the free amine was added the dichloromethane solution of the syn isomer of 2- (fur-2-yl) -2-methoxyiminoacetyl chloride and 5 ml of propylene oxide. After ten minutes, 1.1 g of crystalline solid was filtered off, which was later identified as diphenylmethyl (6R, 7R) -7-amino-3-carbamoyloxymethylceph-3-em-4-carboxylate hydrochloride. The filtrate was washed with 2N sulfuric acid, water, aqueous sodium bicarbonate solution and water and dried over magnesium sulfate and evaporated to dryness to give 2.5 g (30.5%) of the title compound, the physical properties of which were as in the product obtained. by the method described above.

b) Sodium (6K, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate (syn isomer) 20 ml of trifluoroacetic acid was added slowly to a mixture of 5 ml of anisole and 4.7 g (8 mmol) of the syn isomer of diphenylmethyl- (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] ceph-3-em-4-carboxylate which had been cooled in an ice bath. The flask was occasionally shaken over the next ten minutes to ensure complete dissolution of the solid. It was then removed from the ice bath and excess trifluoroacetic acid removed on a rotary evaporator. Trituration of the residue with 5 ml of ethyl acetate gave 3.3 g (94%) of the syn isomer of (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido ceph-3-em-4-carboxylic acid as a solid which was filtered off and washed with diethyl ether.

147683 41

The free acid was dissolved in acetone and a small excess of sodium 2-ethyl hexanoate in acetone (8.0 ml of a molar solution) was added. After the reaction mixture was stirred at 0 ° C for two hours, 2.3 g (73%) of the title salt was filtered off. This was combined with a second portion of 0.8 g of the title salt and purified by washing an aqueous solution (250 ml) with 2 x 100 ml and 1 x 50 ml ether.

The aqueous solution was freeze-dried to give 2.66 g of the syn isomer of sodium (6R, 7R) -3-carbamoyloxymethyl-7- / 2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3 em-4-carboxylate. = + 73.5 ° (c = 1.06 in Me 2 SO), Amax (pH 6 buffer) 274 nm (£ = 16,500). max (nujol) 3450, 3330, 3250 (NH, NH2 and HgO), 1752 (azetidin-2-one) and 1652 and 1600 cm-1 (carboxylate); (MegSO-d 2) 0.24 (d, J 8 Hz, CONH), 2.12 (d, J 2 Hz, furyl C 5 -H), 3.25 and 3.30 (m, furyl C 3 -H and C4-H), 3.44 (broad s, COHH2), 4.34 (dd, J 5 and 8 Hz, C? -H), 4.92 (d, J 5 Hz, C6-H), 5, 15 (q, J 13 Hz, C₂-CHg), 6.07 (s, NOCH₂) and 6.58 (q, J 18 Hz, C₂-H2).

Calcd. For U 2 H 3 N 3 NaO 2 S.0.5H 2 O (455.37): C 42.2 H 3.5 N 12.3 S 7.0 Found: C 42.0 H 3.8 N 12.1 S 7 , 2%.

Example 2 (6R, 7R) -3-Carbamoyloxymethyl-7 (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

A stirring mixture of 75 ml of N, N-dimethylacetamide, 75 ml of acetonitrile, 42 ml (0.3 mol) of triethylamine and 16.40 g (0.06 mol) of (6R, 7R) -7-amino 3-carbamoyloxymethylceph-3-em-4-carboxylic acid was immersed in an ice bath and 10 ml of water was added. The mixture was stirred at 0-2 ° C for 45 minutes, slowly dissolving the solid to give a yellow solution.

Meanwhile, a stirred suspension of 14.99 g (0.072 mol) of phosphorus pentachloride in 150 ml of dry dichloromethane was cooled to 0 ° C and 27.5 ml of N, N-dimethylacetamide was added.

The resulting solution was again cooled to -10 ° C and 12.17 g (0.072 mol) of 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn isomer) was added. The mixture was stirred at -10 ° C for 15 minutes and 35 g of crushed ice was added. The mixture was stirred at 0 ° C for ten minutes and then the lower dichloromethane phase was added over a period of ten minutes to the cephalosporin solution cooled to -10 ° C in accordance with the above, so that the reaction temperature slowly increased to 0 ° C. The mixture was stirred at 0-2 ° C for one hour after which the cooling bath was removed and the reaction temperature was allowed to rise to 20 ° C over one hour. The reaction mixture was then slowly added to 100 ml of 2N hydrochloric acid diluted with cold water (1.15 liters) at 5 ° C. The pH of the biphasic mixture was adjusted to below 2 with 10 ml of 2N hydrochloric acid and the mixture was stirred and cooled again to 5 ° C. The solid which precipitated was filtered, washed with 100 ml of dichloromethane and 250 ml of water and dried in vacuo at 40 ° C overnight to give 22.04 g (86.6%) of the title compound. aD = + 58 ° (c = 1.08 in Me 2 SO); Λmax (pH 6 phosphate buffer) 274 nm (ε = 17,500); Vmax (nujol) 3480, 344o, 3367, 3255 and 3133 (linked NH and NH 2), 2725 and 2590 (CO 2 H), 17.60 (azetidin-2-one) -, 1728, 1712 and 1698 (OCONHg and COOH), 1655 and 1530 cm "1 (CONH); (Me₂SO-d6) 0.25 (d, J 8 Hz; CONH), 2.18 (s, furyl C5-H), 3.28 and 3.4 (m, furyl C4-H and C3-H), 3.42 (s, COHH2), 4.19 (dd, J 8 and 5 Hz; C? -H), 4.80 (d, J 5 Hz; Cg-H ), 5.06 and 5.39 (q, J 13 Hz; Cg-CHg), 6.09 (s; NOCH ^), 6.44 (collapsed ABq; Cg-H;?) 7.99 (0, 03 mol CH3CON (CH3) 2).

Example 3 a) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-hydroxymethylceph-3-em-4-carboxylic acid monoquinolinium salt monohydrate

A stirred solution of 18.45 g (30 mmol) (6R, 7K) -7- (R-5-amino-5-carboxypentanamido) -3-hydroxymethylceph-3-em-4-carboxylic acid monocalcium salt in 93 ml water was cooled to 0-5 ° C (ice / water bath) and treated with a solution of 5.19 ml (45 mmol) of benzoyl chloride in 63 ml of acetone over 25 minutes. The pH of the reaction mixture was maintained at 8.5 µl 0.1 by controlled addition of ca.

100 ml of 30% s / v aqueous tricalcium orthophosphate. The mixture was stirred for a further five minutes and covered with 150 ml of ethyl acetate, then the pH was reduced to 5.6 with orthophosphoric acid. The layers were separated and the aqueous portion was washed with an additional 2 x 300 ml of ethyl acetate. The combined washings were extracted with 200 ml of water. The aqueous phase was combined with the washing liquids and the liquids thus combined were diluted with 600 ml of water and covered with 600 ml of ethyl acetate and the pH of the stirred mixture was adjusted to 2.0 with orthophosphoric acid. The organic layer was separated and 10.64 ml (45 mmol) of quinoline added in 25 ml of ethyl acetate with stirring to give a white precipitate. The aqueous portion was extracted with an additional 3 x 300 ml of ethyl acetate and these were added to the quinoline-containing suspension. The mixture was stirred for one hour at ca. 18 ° C and then concentrated in vacuo to ca. 500 ml. 900 ml of ether were added with stirring and after 30 minutes the solid was collected by filtration, washed with 5 x 100 ml of ether and dried in vacuo (1 mm Hg) to give the title compound as a white powder in an amount of 19.20 g (104.1%), aj8 = + 78 ° (c = 1.00 in dioxane). kmax (pH 6 buffer) 258 nm (inflection, E ^ cjn = 185). IR and NMR data confirmed the structure as the title compound structure containing ca. 15% lactone impurity and trace amounts of ether and ethyl acetate.

b) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid monoquinoline salt 4.24 g of the product of Example 3 a), corresponding to 7 mmol, treated with 100 ml of dry dioxane in which the product partially dissolved. To the stirred mixture was added 2.90 ml (24.5 mmol) of trichloroacetyl isocyanate. The resulting solution was stirred for 30 minutes and then clarified by filtration and evaporated in vacuo to a yellow foam. This dissolved in ca. 10 ml of acetone and poured into ca. 100 ml stirred isopropyl ether. The resulting white precipitate was collected by filtration and dried in vacuo (1 mm) to give 6.26 g (147.8%) of the title compound as a white powder. NMR data confirmed that the structure was the structure of the title compound and also showed the presence of lactone (about 22%), isopropyl ether (0.75 mol), dioxane (0.2 mol) and a small amount of acetone.

c) (6R, 7E) -7-Amino-3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid

A solution of 4.77 g of the product of Example 3b), corresponding to 6 mmol, in 40 ml of dry methylene chloride under nitrogen was cooled to ca. 10 ° C with stirring. 2.20 ml (27.3 mmol) of pyridine and then 2.10 ml (17.4 mmol) of dichlorodimethylsilane were added and the brown solution was stirred at ca. 17 ° C for twenty minutes and then cooled to -17 ° C. Phosphorus pentachloride (1.355 g (6.5 mmol)) was added and the mixture was stirred at ca. -16 ° C for two hours. 0.81 ml (10 mmol) of pyridine was added and the mixture was added to methanol (13 ml + 2.5 ml washing liquids, pre-cooled to -35 ° C) at such a rate that the temperature of the stirred solution did not exceed -10 ° c. The stirring solution temperature was allowed to reach + 9 ° C over 25 minutes, then 11 ml of water was added and the pH of the mixture was adjusted from 0.3 to 3.8 with ammonia water (density 0.880). The resulting two-phase mixture containing a precipitated solid was refrigerated in the refrigerator for one hour and then filtered. The solid was washed successively with 12 ml of 50% aqueous methanol, 10 ml of methanol and 5 ml of methylene chloride and then dried in vacuo (1 mm) to give 1.22 g (25.6%) of the title compound. as a cream colored powder, α D = + 44 ° (c = 1.02 in MeoSO); \ max (pH 6 buffer) 240 nm (b} + = 133), 263 nm (B + = 140). NMR data confirmed that the structure of the product was the title compound.

d) (6R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxy-iminoacetamido7-ceph-3-em-4-carboxylic acid (syn isomer) 4.5 g Phosphorus pentachloride (21.5 mmol) was dissolved in 90 ml of dry methylene chloride and cooled to -15 ° C with stirring. 9 ml of Ν, Ν-dimethylacetamide was slowly added keeping the temperature below -7 ° C -10 ° C and the mixture stirred for ten minutes. Then, 3.66 g corresponding to 21.5 mmol of the syn isomer of 2- (fur-2-yl) -2-methoxyiminoacetic acid was added and the mixture was stirred at -15 ° C for 15 minutes. Carefully add 18 g of crushed ice so that the temperature of the mixture did not exceed -7 ° C. Then the mixture was stirred for ten minutes and the organic part was separated and added dropwise to a solution of 7.52 g corresponding to 18 mmol, (6R, 7R) -7-amino-3-trichloroacetylcarbamoyloxymethylceph-3-em-4-carboxylic acid in 90 ml of dry methylene chloride and 5.5 ml (40 mmol) of triethylamine, pre-cooled to -10 ° C. The acid chloride solution was added over 20 minutes keeping the temperature of the reaction mixture between -10 and -8 ° C. Then the mixture was stirred for 80 minutes during which time the temperature was allowed to rise to + 3 ° C and 6 ml of methanol was added. After a further five minutes of stirring, the solution was extracted with 2 x 120 ml of 3% aqueous sodium hydrogen carbonate and 250 ml of water. The combined extracts were allowed to stand at ca. 20 ° C for 3 1/2 hours and then washed with 100 ml of ethyl acetate and acidified to pH 1.5 with concentrated hydrochloric acid. The resulting deposited oil was extracted with 2 x 300 ml of ethyl acetate. The combined organic parts were washed with 2 x 100 ml of water, dried over MgSO 4 and evaporated in vacuo to 7.1 g of yellow solid which was stirred with 150 ml of ether, filtered and dried in vacuo at 1 mm Hg to give the title the compound of 5.20 g (68.2%) of a light yellow solid with λ max (pH 6 buffer) 275 nm (E + = 385).

IR and NMR data confirmed that the structure of the product was the structure of the title compound containing traces of ether.

Example 4 (6R, 7R) -3-Carbamoyloxymethyl-7- / 2-methoxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

By the same procedure as described in Example 2, 888 mg (50 $) of the title compound was prepared from 1.09 g (4 mmol) (6R, 7R) -7-amino-3-carbamoyloxymethylceph-3-em. 4-carboxylic acid and 923 mg (4.8 mmol) of the sodium isomer of sodium 2-methoxyimino-2- (thien-2-yl) acetate. The properties of the compound are: mp 157-163 ° C, αβ = 57.3 ° (c = 1.0 in dioxane), EpAC 0.8, solvent system A (see notes following Table 1), Xmax (pH 6 buffer) 262 , 5 nm (ε = 15,550) and inflection at 235 nm (e = 10,350), δ ir (DMSO-dg) 0.20 (d, J 8 Hz, NH), 2.29 (dd, J 2 and 5 Hz) , thienyl C ^-H), 2.7-2.9 (m, thienyl Cg-H and C4-H), 3.40 (s, CONH₂), 4.13 (dd, J 5 and 8 Hz, C7 -H), 4.75 (d, J 5 Hz, Cg-H), 5.01 and 5.34 (AB-q, J 13 Hz, Cg-CHg), 6.08 (s, NOCH and 6.42 (collapsed AB-q, C2-H2), Vmax (nujol) 3700 to 2100 (COOH), 3480, 3440, 3365 and 3255 (NH and NH2), 1760 (azetidin-2-one), 1722 ( COOH), 1709 (OCNH 2), 1652 and 1530 cm -1 (amide).

Example 5 (6R, 7R) -3-Carbamoyloxymethyl-7- [5- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid (syn isomer) 1.5 ml ml Ν-Dimethylacetamide was added to a solution of 750 mg (3.6 mmol) of phosphorus pentachloride in 15 ml of anhydrous dichloromethane cooled to -10 ° C. After ten minutes, 612 mg, 3.6 mmol, of the syn-isomer of 2- (fur-2-yl) -2-methoxyiminoacetic acid was added to the resulting suspension, which quickly became a clear solution and stirred at 10 ° C for 10 min. 15 minutes. 3 g of ice was added. After ten minutes, the layers were allowed to separate into a dropping funnel. The organic phase was slowly transferred, over five minutes, to a solution cooled to -10 ° C of 1.05 g (3 mmol) (6R, 7R) -7-amino-3-chloroacetylcarbamoyloxymethylceph-3-em-4- carboxylic acid in 15 ml of dichloromethane containing 0.9 ml, 6.5 mmol, triethylamine. After 40 minutes, 1 ml of methanol was added and five minutes later the reaction mixture was extracted twice with 150 ml of 3% aqueous sodium bicarbonate solution. The aqueous extract was washed with 25 ml of ethyl acetate and left at 20 ° C for four hours. The solution was washed twice with ethyl acetate, acidified with 2N hydrochloric acid and extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate, decolorized with activated charcoal and evaporated in vacuo to 1.15 g, 87%, of a light yellow solid. This was washed with diethyl ether and filtered off to give 0.91 g, 71% of the title compound. Amax (pH 6 buffer) 274 nm (e = 17 * 300). The product's IR and NMR spectra matched the spectra for an authentic sample of the compound.

Example 6 (6R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

Method I

750 ml of acetone was cooled to 0 ° C and treated with 28.8 ml, 240 mmol of trichloroacetyl isocyanate and the solution was again cooled to 0 ° C. 45.6 g, 120 mmol, (6R, 7R) -7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-3-hydroxymethylceph-3-em-4-carboxylic acid (the syn isomer) were added to the stirred isocyanate solution in portions over five minutes in such a way that the reaction temperature did not exceed 6 ° C. The yellow solution was stirred for a further 15 minutes and 4.5 ml of methanol was added. The solution was concentrated to 60 ml and the concentrate dissolved in 750 ml of methanol. 45.3 g (540 mmol) of sodium bicarbonate was added in 600 ml of water followed by 4.5 g of activated charcoal and the resulting suspension was stirred at room temperature for two hours. The charcoal was removed by filtration through diatomaceous earth and the pale yellow filtrate was adjusted to pH

4.5 by the addition of dilute hydrochloric acid. The solution was concentrated to half the volume under reduced pressure and an equal amount of water was added. The pH was adjusted to 2.0 with dilute hydrochloric acid and the product isolated by filtration, washed with 3 x 150 ml of water and dried at 40 ° C for 16 hours in vacuo to give 37.46 g corresponding to 73.5% of the title compound, ap® = + 63.7 ° (c = 1.0 in 0.2 M pH 7 phosphate buffer). Amax (pH 6 phosphate buffer) 274 nm (e - 17,600).

IR, NMR and microanalytical data confirmed that the structure was the title compound.

Method II

48 167683

A slurry of 3.81 g, 9.55 mmol, of the syn isomer of (6R, 7R) -7- / 2- (fur-2-yl) -2-methoxyiminoacetamido7-3-hydroxymethylceph-3-em-4 -carboxylic acid in a mixture of 70 ml of dichloromethane and 25 ml of tetrahydrofuran at 5 ° C was treated with 2.5 ml, 25 mmol, dichloroacetyl isocyanate. The reaction mixture was then treated in the same manner as described above under Method I to yield 3.36 g, corresponding to 83.0% of the title compound. = + 63®, λmax 273.5 nm (e = 17,800). IR and NMR spectra were like the spectra of the product produced by Method I.

Method III

A slurry of 19.05 g, 50 mmol, of the syn isomer of (6R, 7R) -7- / 2- (fur-2-yl) -2-methoxyiminoacetamido7-3-hydroxymethylceph-3-em-4 carboxylic acid in 250 ml dry acetonitrile was treated at a temperature between 5 and 10 ° C with 6.33 ml, 75 mmol, chlorosulfonylisocyanate in 80 ml acetonitrile.

The reaction mixture was stirred at temperatures between 0 and 5 ° C for ten minutes and 50 ml of water was added. The mixture was stirred at ca. 20 ° C and after twenty minutes a white crystalline solid separated. Evaporation and filtration gave 18.17 g (85.7%) of the title compound, ctD = + 62.5®; ν max 273.5 nm (ε = 17.820). IR and NMR spectra were similar to those found for the product of Method I. A next yield of 1.88 g, 8.6%, of the product of similar constants was obtained by evaporation of the parent liquids.

Example 7

Form I of the syn isomer of sodium (6R, 7R) -3-carbamoyloxymethyl-7-72- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate

Method I

49 g of the syn isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid in a mixture of 400 ml of Ν, Ν-dimethylacetamide and 1 liter of acetone were treated with 40 g of sodium 2-ethyl hexanoate in 200 ml of acetone. The mixture was seeded and stirred at room temperature for 1 1/4 hour. The product was filtered off and washed with 500 ml of acetone, then slurried with 3 x 300 ml of acetone and finally slurried with ether to give 101.4 g (92.5%) of the title compound which, after reaching equilibrium, in the atmosphere contained 0.65 molar equivalents of water. The product has α D = + 61 ° (c = 0.5 in pH 4.5 phosphate buffer) and λ max 273 nm (E + cm = 412) (HgO).

IR and NMR data confirmed that the structure was the titled compound structure and the IR spectrum indicated that the compound was salt of Form I.

Method li

The method of method I was repeated with the difference that the cephalosporic acid was initially dissolved in a mixture of N, N ~ dimethylformamide and industrial methylated ethanol instead of a mixture of Ν, Ν-dimethylacetamide and acetone to give the title compound in a yield of 80% with similar properties to the product of Method I. The infrared spectrum showed that the compound was the salt of Form I.

Method III

4.24 g, 10 mmol, of the syn isomer of (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4 Carboxylic acid was dissolved in 20 ml of Ν, Ν-dimethylacetamide which had been dried over a molecular sieve material ("Linde 4A") for 24 hours. To this was added a solution of 2.0 g, 12 mmol, sodium 2-ethyl hexanoate, was recrystallized from dioxane and dried over phosphorus pentoxide, in 80 ml of ethyl acetate which had been dried over a molecular sieve material ("Linde 4A") for 24 hours. The solution was stirred in a closed vessel for about 15 minutes until crystallization began, then cooled to The product was filtered, washed with about 100 ml of dry ethyl acetate and transferred, in a state still wet by the latter solvent, to an oven where it was dried at 20 ° C in vacuo over phosphorus pentoxide overnight; thus obtained 3 »84 g * 87% of the title compound.

The product's IR and NMR spectra matched the corresponding spectra for an authentic sample of the fabric.

Example 8

Form II of the syn isomer of sodium (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate

Method I

0.2 g of charcoal was added to a solution of 4.00 g, 9.42 mmol, of the syn isomer of (6R, 7R) -3-carbamoyloxymethyl-7- / 2- (fur-2-yl) -2- methoxyiminoacetamido / ceph-3-em-4-carboxylic acid in a mixture of 132 ml of acetone and 1.33 ml of water. The suspension was stirred for 30 minutes and filtered through a diatomaceous earth bed which was washed with 10 ml of acetone. A filtered solution of 1.66 g, 10 mmol, 2-ethylhexanoate in 20 ml of acetone was added over one hour to the stirred filtrate. The resulting suspension was stirred for a further ten minutes and the white solid was filtered off, washed with 2 x 25 ml of acetone and dried in vacuo to 4.06 g, 93.0%, of the title compound at = + 60 ° ( c = 0.91 in H2 O). λ max (H 2 O) 274 nm (ε = 17,400).

Calcd for C16 H15 N4 Na08S, 0.7 HgO (459.0): G 41.8, H 3.6, N 12.2, Na 5.0, S 7.0, H2 O 2.7%; found C 41.0, 41.2; H, 3.45; 3.6; N, 12.3, 12.4; Na 5.2; S, 6.6, 6.85; H2 O 2.7, 2.7. Purity by HPLC: 99.4%. The NMR spectrum of the product was similar to the spectrum of an authentic sample and the IR spectrum showed that the product was salt of Form II.

147683 51!

Method II

16 »98 g, 40 nunol, of the syn isomer of (6R, 7R) -3-carbamoyl-o: xymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em. 4-carboxylic acid was added to a stirring mixture of 333 ml of acetone and 8.5 ml of water. After treatment with charcoal and filtration of this solution, 7.32 g, 44 mmol sodium 2-ethyl hexanoate in 85 ml of acetone was slowly added over one hour. The reaction mixture was stirred for 15 minutes and filtered and the product washed with 2 x 65 ml of acetone and dried in vacuo at 20 ° C overnight to give 17 »95 g, 98.5% of the title compound containing 0.5 mole H ^ O. The NMR spectrum of the product matched an authentic sample, and the IR spectrum indicated that the product was salt of Form II.

Example 9

Form III of the syn isomer of sodium (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate 40 g of the syn isomer of sodium (6R, 7R) -3-carbamoyloxy-methyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate was dissolved in 20 ml of water. 20 ml of industrial methylated ethanol and 160 ml of dioxane were added and the solution was filtered and then set aside at 4 ° C to crystallize. The very white needle-shaped crystals were filtered off, washed with 100 ml of dioxane and transferred to a state where they were still wet with dioxane in an oven and dried at 20 ° C in vacuo overnight; 13.96 g, 78.5% of the title compound were obtained. The product's IR and NMR spectra matched those of an authentic sample.

Example 10 52 147683

Form IV of the syn-isomeric sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate

Samples of Form I and Form III of the syn isomer of sodium 3-carbaoyloxymethyl-7- / 2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate prepared by Method III in Example 7 and Example 9, respectively, were exposed to humidity (75% relative humidity) for three days to give the title compound. The product's IR and NMR spectra matched those of an authentic sample. Water analysis by Karl Fischer yielded 4.0 and 3.85%, respectively (1 mole of H 2 O is equivalent to 3.9%).

Example 11 a) t-Butyl (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate (synisomer)

A suspension of 4.4 g of the syn isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid in 200 ml of dry methylene chloride was treated with 6.6 ml of Ot-butyl-N, N * -dicyclohexylisourea to give a pale yellow solution. After 24 hours at 23 ° C, starting material was left and an additional 3.3 ml of the isour substance was added. After 48 hours, the mixture was filtered and the filtrate was evaporated under reduced pressure. The resulting material was slurried with ether / ethyl acetate to remove residual dicyclohexylurea. The filtrate was washed with saturated aqueous sodium bicarbonate and water, then dried and evaporated to a foam in an amount of 5.2 g. Chromatography on silica gel with toluene / ethyl acetate 2: 1 as solvent gave 3.9 g of the title compound pale yellow foam, \ max (ethanol) 275.5 nm (ε = 18,400).

IR and NMR data confirmed that the structure was the title compound.

B) t-Butyl (IR, 6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate-1-oxide (syn isomer) 0.3 ml of t-butyl hypochlorite was added to a solution with vigorous stirring of 0.98 g of t-butyl (6R, 7R) -3-carbamoyloxy-methyl-7- / 2- (fur-2 -yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylate in 25 ml of pyridine and 1 ml of water at -45 ° C. After stirring for two minutes, 1 ml of 2N sulfuric acid was added to the solution and the mixture was immediately poured into 100 ml of 20% aqueous orthophosphoric acid. The solution was extracted with 2 x 100 ml of ethyl acetate and the combined organic extracts washed with 100 ml of aqueous NaHCO 3 and 100 ml of water and then dried over MgSO 4 and concentrated in vacuo.

The crude product was chromatographed on silica gel preparative plates with ethyl acetate as the eluent, and the minor component of the S-oxide preceded the R-oxide. Extraction of the slowest component with ethyl acetate gave 0.27 g (27 $) of the title compound.

IR and NMR data confirmed that the structure was the title compound.

c) (IR, 6R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em-4-carboxylic acid 1-oxide (syn isomer ) 0.42 g of the syn isomer of t-butyl (1R, 6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em -4-Carboxylate-1-oxide was dissolved in 5 ml of trifluoroacetic acid and stirred at room temperature for eight minutes. The solution was evaporated to a red oil in vacuo, dissolved in 5 ml of ethyl acetate / acetone 1: 1 and added dropwise with stirring to 50 ml of petroleum ether (bp 6o-80 ° C). The deposited solid was collected and dried in a desiccator. The crude product was slurried with ethyl acetate and the liquid phase decanted and added dropwise to 50 ml of petroleum ether (bp 60-80 ° C) to give 150 mg, $ 40, of the title compound as a colorless solid. \ max (0.25N NaHCO3) 263.5 nm (ε = 15,000) and 281 nm (ε = 13,700). max (nujol) 1799 (β-lactam), 1727 and 1716 147683 54 (COOH and OCONH2) f 1684 and 1538 (CONH) and 1060 and 1050 cm -1 (s -> 0). C (Me2SO-cl6) values include 0.02 (d, J 8 Hz, CONH), 4.17 (dd, J 4 and 8 Hz, 7-H), 4.99 (d, J 4 Hz, 6-H) and 6.09 (s , N-OCH 3).

Example 12 (1S, 6R, 7E) -3-Carbamoyloxymethyl-7- {2- (fur-2-yl) -2-methoxy-iminoacetamido7-cepi-3-em-4-carboxylic acid 1-oxide (syn isomer )

To a stirred solution of 2.59 g of the syn isomer of sodium (6R, 7R) -3-carbamoyloxymethyl-7- / 2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em -4-carboxylate in 25 ml of water was added 1.93 g of sodium metaperiodate. The solution was stirred for 30 minutes at room temperature and then acidified by dropwise addition of 2N aqueous hydrochloric acid. The resulting precipitate was collected, washed successively with water, ethanol and ether and then dried in vacuo to give 1.63 g of the title compound as a white powder, = + 113 ° (c = 0.86 in dimethyl sulfoxide). ). Xmax (pH 6 buffer) 264.5 nm (ε = 17,200) and 279 nm (ε = 15,600).

> 7max (nujol) 1770 (β-lactam), 1740 and 1716 (COOH), 1688, 1654, 1589 and 1530 (CONH and OCNH2) and 1030 cm -1 (S-O). ^ (Me2SO-d6) 0 , 60 (d, J 8 Hz, NH), 2.11, 3.19, 3.31 (multiplets, furyl protons), 4.08 (q, J 5 and 8 Hz, C-7H), 4, 87 and 5.45 (ABq, J 13 Hz, CH 2 CONH 2), 4.96 (d, J 5 Hz, C-6H), 6.08 (s, OCH 3), 6.10 and 6.42 (ABq, J 18 Hz, C-2CH 2).

EXAMPLE 13 (1R, 6R, 7R) - and (1S, 6R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido7-ceph-3-em] 4-carboxylic acid 1-oxide (syn isomers)

A solution of 1.93 g of sodium periodate in 10 ml of water was added to a stirred solution of 2.59 g of sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2 -methoxyiminoacetamido7-ceph-3 ~ em-4-carboxylate in 25 ml of water. Stirring was continued for 30 minutes at room temperature and the solution was then acidified with 2 ml of 2N HCl. The precipitated 1S oxide was filtered off, washed with ethanol (5 ml) and ether (20 ml) and dried in a desiccator to give 1.5 g of colorless solid, aD = + 110 ° (c = 1e Me 2 SO). described in Example 12.

The mother liquors were saturated with sodium chloride and filtered and the filtrate was extracted with 2 x 100 ml of ethyl acetate. The organic extracts were combined, dried over MgSO 4 and concentrated in vacuo to give a yellow solid. The crude product was washed with acetone and the insoluble material removed by filtration. The filtrate was evaporated to dryness and the acetone washed repeatedly to give 380 mg of the 1R oxide with aD = -88 ° (c = 1 in Me2S0), ΙΓ values (Me2S0-dg) similar to those listed in Example II.

Example 14 a) Diphenylmethyl (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-phenoxyiminoacetamido7-ceph-3-em-4-carboxylate (syn isomer)

A solution of 7 »75 g (0.382 mol) of DL-dicyclohexylcarbodiimide in 50 ml of dry dichloromethane was added over 10 minutes to a solution of 13» 7 g »0.312 mol, diphenylmethyl (6R, 7R) -7-amino -3-carbamoyloxymethylceph-3-em-4-carboxylate and 8.8 g, 0.382 mol, of the syn isomer of 2- (fur-2-yl) -2-phenoxyiminoacetic acid in 200 ml of dry dichloromethane at 0 ° C. After 45 minutes, a solid (presumably Ν, Ν'-dicyclohexylurea) was filtered off and the filtrate washed with aqueous sodium bicarbonate solution and water, dried over magnesium sulfate and evaporated on a rotary evaporator. The residue was chromatographed on a column of 1 kg of silica gel. Impurities less polar than the desired product were eluted with 1 liter of dichloromethane and 1 liter of acetone / dichloromethane 2:98 and 4 liters of acetone / dichloromethane 5:95. Fractions eluted with acetone / dichloromethane 10:90 and acetone / dichloromethane 15:85 were evaporated to a gum in an amount of 11 g and triturated with diethyl ether to give 8.35 g ($ 41) of a solid. This was filtered off and further purified by crystallization from aqueous ethanol to give 7.6 g of the title compound, mp 143-146 ° C, α D = + 48 ° (c = 1.0 in Me₂SO). Xmax (ethanol) 273 nm (ε = 18.700). Xinfl. (ethanol) 271 nm (ε = 17,600) and Xmax (ethanol) 254 nm (ε = 16,500).

IR, NMR and microanalytical data confirmed that the structure was the title compound.

b) Sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-phenoxyiminoacetamido7-ceph-3-em-4-carboxylate (syn isomer) 30 ml trifluoroacetic acid was added over ten minutes to an ice-cooled mixture of 8 ml of anisole and 7.4 g, 11.4 mmol, of the syn isomer of diphenylmethyl- (6R, 7R) -3-carbamoyloxymethyl-7- / 2- (fur-2 yl) -2-fenoxyiminoacetamidoT-ceph-3-em-4-carboxylate. After another five minutes at 0 ° C, the dark solution was gently poured into a mixture of a saturated aqueous solution of sodium bicarbonate and ether acetate. The aqueous solution was separated and treated with activated charcoal. The ethyl acetate layer was washed with water and the aqueous wash liquid combined with the sodium bicarbonate extract and acidified with concentrated hydrochloric acid. This acidic solution was extracted with a mixture of ethyl acetate and diethyl ether which was then washed five times with water, dried over magnesium sulfate and evaporated in vacuo. The residue was washed with diethyl ether and diisopropyl ether to give 4.5 g (82 $) of the cephalosporic acid as a solid.

147683 57

This acid was dissolved in 150 ml of ethyl acetate and a solution of sodium 2-hexylhexanoate in ethyl acetate (10 ml containing 10 mmol) was added. The solution was cooled in an ice bath and stirred for 1 1/2 hours during which time 2.84 g of the product crystallized while leaving unchanged acid in solution (1.1 g recovered by precipitation with petroleum ether, bp 60-80 ° C). The solid was filtered to give the title compound.

Physical constants for the title compound are given in Table 1 below.

Example 15 a) Diphenylmethyl (6R, 7R) -3-carbamoyloxymethyl 7- (2-methoxy-imino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn-isomer) 1.83 g, 3 mmol, diphenylmethyl (6R, 7R) -7-amino-3-harbamoyl-oxymethylceph-3-em-4-carboxylate-p-toluenesulfonic acid salt was added to a mixture of 50 ml of saturated aqueous sodium bicarbonate solution and 100 ml of dichloromethane. The mixture was shaken to dissolve the solid and the organic layer was separated, washed twice with water, dried over sodium sulfate and concentrated under reduced pressure to ca. 15 ml. This solution was cooled to 0 ° C and solutions of 824 mg, 4 mmol, DL-dicyclohexylcarbodiimide in 10 ml of dry dichloromethane and 716 mg, 4 mmol of the syn-isomer of 2-methoxy-mino-2-phenylacetic acid were added in 10 ml. dry dichloromethane. The reaction mixture was stirred at 0 ° C for 70 minutes, then filtered, washed with 2N sulfuric acid, water, saturated aqueous sodium bicarbonate solution, water and brine, dried over sodium sulfate and evaporated to 2.05 g of a yellow solid. This was dissolved in 25 ml of hot methyl acetate which was cooled and the dicyclohexylurea was filtered off. Addition of diisopropyl ether to the filtrate caused the precipitation of a solid which was filtered, stirred and washed with diisopropyl ether, filtered and finally washed with ethyl ether; There was thus obtained 1.10 g (61.5%) of the title compound, mp 178-182 ° C. aD = + 22.5 ° (c = 1 in CHCl 3). \ max (ethanol) 258.5 nm (ε = 18,500), Xinfl. (ethanol) 295 nm (ε = 3120).

IR, NMR and microanalytical data confirmed that the structure was the title compound.

b) (6R, 7R) -3-Carbamoyloxymethyl-7- (2-methoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

Treatment of the product of Example 15 a) in accordance with the method described in Example 1b) provided 75% of the title compound and was purified by trituration of the crude product with 10 ml of ethyl acetate followed by washing twice with stirring with diethyl ether. , 25 ml and 10 ml respectively.

Physical constants for the title compound are listed in Table 1 below.

Example 16 a) Diphenylmethyl (6 R, 7 R) -3-carbamoyloxymethyl-7- (2-phenylimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

The procedure of Example 15 a) was repeated with the difference that the syn isomer of 2-phenoxyimino-2-phenylacetic acid was used instead of the syn isomer of 2-methoxyimino-2-phenylacetic acid and a red sticky foam was obtained. This crude product was crystallized from aqueous acetone to give 1.3 g of a red solid which was washed twice with stirring with diethyl ether; there was obtained 675 mg (34%) of the title compound, mp 138-140 ° C. aD = + 44.5 ° (c = 1 in CHCl3). Xmax (ethanol) 264 nm (ε = 12,400), \ infl. (ethanol) 268 and 281 nm (ε = 16,850 and 14,000); the product was a white solid. The residue obtained after evaporation of the mother liquors and washings was crystallized from ethanol to give an additional 431 mg (17%) of the title compound in two yields which were washed with diethyl ether with stirring.

IR, NMR and microanalytical data confirmed that the structure of the product was the title compound.

B) Sodium (6R, 7R) -3-carbamoyloxymethyl-7- (2-phenoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn-isomer)

Treatment of the product of Example 16 a) by the method described in Example 1 b) gave the crude cephalosporic acid and it was stirred with ethyl acetate and saturated aqueous sodium bicarbonate solution. The resulting precipitate was filtered off and washed with acetone and ether to give the title compound (66%) having the physical properties shown in Table 1 below.

Example 17 a) Diphenylmethyl (6R, 7R) -3-carbamoyloxymethyl-7- / 2-cyclo-pentyloxyimino-2- (fur-2-yl) -acetamido7-ceph-3-em-4-carboxylate (syn-isomer) )

The procedure of Example 15 a) was repeated with the difference that the syn isomer of 2-cyclopentyloxyimino-2- (fur-2-yl) acetic acid was used instead of the syn isomer of 2-methoxyimino-2-phenyl acetic acid. and 1.77 g of a foam was obtained which was triturated with ethyl acetate to give 1.30 g (67%) of the title compound as a pale yellow solid, mp 102-108 ° C. <* D = + 12.5 ° (c = 1 in CHCl 3). Xmax (ethanol) 278 nm (e = 16,650).

b) (6R, 7R) -3-Carbamoyloxymethyl-7- / 2-cyclopentyloxyimino-2- (fur-2-yl) -acetamido7-ceph-3-em-4-carboxylic acid (syn isomer)

Treatment of the product of Example 17 a) in accordance with the method described in Example 1 b) gave the crude cephalosporic acid which precipitated from ethyl acetate with diisopropyl ether; thereby winning the title compound (55%) with physical constants as shown in Table 1 below.

Example 18 60 147683 a) Diphenylmethyl (6R, 7R) -3-carbamoyloxymethyl-7- (2-ethoxy-imino-2-phenylacetamido) -ceph-3-em-4-carboxylate (syn isomer)

The procedure of Example 15 a) was repeated except that the syn isomer of 2-ethoxyimino-2-phenylacetic acid was used instead of the syn isomer of 2-methoxyimino-2-enylacetic acid. Crystallization of the crude product from methanol afforded 1.30 g (53%) of the title compound in three yields, mp 199-202 ° G, α D = + 9.7 ° (c = 1 in dioxane). Xmax (ethanol) 259 nm (ε = 20,000). \ Infl. (ethanol) 295 nm (ε = 3700).

b) (6R, 7R) -3-Carbamoyloxymethyl-7- (2-ethoxyimino-2-phenylacetamido) -ceph-3-em-4-carboxylic acid (syn isomer)

Treatment of the product of Example 18 a) in the manner described in Example 1b) gave the crude cephalosporic acid and it was triturated with 3 ml of ethyl acetate, filtered and washed with stirring with 5 ml of ethyl acetate and then with 2 x 10 ml of diethyl ether. ; 413 mg (64%) of the title compound was obtained as a white solid. An additional amount of 180 mg (27%) crystallized from the filtrates and filtered off and washed with diethyl ether. Physical constants for the title compound are shown in Table 1 below.

Example 19 a) Diphenylmethyl (6R, 7R) -3-carbamoyloxymethyl-7- [2-t-butoxyimino-2- (thien-2-yl) -acetamido7-ceph-3-em-4-carb oxylate (syn isomer)

The procedure of Example 15 a) was repeated except that the syn isomer of 2-t-butoxyimino-2- (thien-2-yl) acetic acid was used instead of the syn isomer of 2-methoxyimino-2-phenylacetic acid. 61 acid. The crude product was purified by stirring a suspension in 2 x 25 ml of diisopropyl ether and the title compound was watered as a quite pink solid in an amount of 1.90 g (73¾), mp 148-152 ° C. aD = + 8.5 ° (c = 1 in CHCl 3). kmax (ethanol) 262 and 282 nm (ε * 14,500 and 13,200).

b) Sodium (6R, 7R) -3-carbamoyloxymethyl-7- / 2-t-butoxyimino-2- (thien-2-yl) -acetajnido7-ceph-3-em-4-carboxylate (syn isomer) )

Treatment of the product of Example 19 a) in the manner described in Example 1b) gave the cephalosporic acid as a gum which solidified upon trituration with diisopropyl ether; 1.20 g (94¾) of a crude product were obtained. A portion of 811 mg (1.68 mmol) of this acid and 282 mg (1.68 mmol) of 2-ethyl hexanoate was stirred in 5 ml of n-butanol at 20 ° C for ten minutes and at 0 ° C for twenty minutes. The resulting yellow solid was filtered off and washed with cold n-butanol (3 ml) and diisopropyl ether (7 ml) to give 495 mg (58¾) of the title compound with the physical constants shown in Table 1.

147083 62 i o tn v o O co o <-l o ©! © I I ©

Q iH r — t iH

►TJ O O O Γν CO CO CM O CM -fr 00 O

g tom tncM mm n- to m cm m cm g © in © in votn © in © m © m

ø i — li — I i — I »-) Η H r-I r-l r-l iH r — I ι — I

* £ m O O O

O 'I r-l O I iH O I

_, q γν i— pn r-

© I O r-l.-I ® ri H

2 κ: o <f © £ <o M -ia r-ι cm σ cm σ »I oo 'r *" p * · o © I g rU rl rl r-ι rl © β n -H + j H r - ^ in o O mmo, <, _} © vo <t ro © in V, 7 7 ^ n. Rr ^ r «-r- pi, g CO. Rl rl i — liHr-li- | II w K OO OO o ®oo SO pn P-. © r-ι O °° p ^ r> SOO CO CM CM CM rl «O fa * 2 ** g r-. Ro ro ro m cm ro 00 ns 2ί ~ Λ o tn il ^ § lei i 1 3., <2 OjO O oooo H | ^ Wj LT \ C * iA o 00 in is ro <f $ £ <f <f Η! Φ j —I --- 57¾ «« ft! * A'Å ^ -g «| 2 i / -s rs o Ο Ο O / - \ Xv ^ | ti ooo oooo o oo 5 oj ooo o © cm cm cm mo wj ^ j - o Γ" (Ti CM · > ·>> ·· O © cjlni ni ω ► * - · n- ro p- σ> ·> - p 1 9 wO · m © ro cor-ir-i r-ι r-ι tn o O rl rl rl rl ^ ι-l il r1 * I v / in in tn

Ti f o s oo o o m © o t n σ «cm σ υ i μ p E σ> γν © © © ro p- tn © ro ^ I nn n cm cm cm cm cm cm cm QJ I, _____ © I 1" _ _

(ti i t n I OO

VJ I O!> 1 "i ffl XS S _J Opj ·. I-l / -V 0 <rv ri N o n O r-N I — I / —s

m I N_r v [<l) r-N · <J · PQ * · · <1 · <1 · CQ

u I O S Ό s r-l N »r r-l v— 'I — I I — I i — I' 1 — i ^ i — 1 v— '

φ | <ΗΤ3 (U

| Cu Ai-H + J

φ I & ^ Q. S t n

Ai I —j —------

.¾ in CO t3 O

55 s-h o ^ -sod o / -s o s. {HE σ o © tti CM o <t o ro ro (C Q.S roto tny h w © w © a mr>

1 O tor-v CM O CM CM o. ISLAND

CHOJ SOS SUS

ΰΑΌ ^ 'r' 'r' WWW

/ n '/ -s rs

. © © © © O O

cn. w w w w © ©

C, No. V

«H <j m © γν. οοσ URr-lr-lr-l r — l r-l r-l 147683 63 Μ cη - • η Ο Ν

-Μ Ν W

(tf OfS γ ~ · Οι Γν • Η <J · {301 ® Ο I-} ΟΟ Η ο / -ν CM θ 'ΓντΊΝ .P * ·' ti Α · · νΟ <J 3 ιή οό5 α ^ Τ οό ^ ΦΝ ^ Χ) · Η Μ -Μ Ιυ || ωε 00 ΙΑ CM CM II Ν I '_ ^ 2? Ti 09 σχη ο <ν η σ' σ 'ih ro <too <ro • H ^ fi, i «i · T · · · ···

W <--- ΗΝΠ Γ0 N N H CM (N Π Π CMCH CM CM

CM N

K | li O ro o r * - sf co Γ ”CMCM ONCO Ό

• o cm O CO in 00 OCO © CO rM

co CQ ,, ··. , ·· ·· ·

O <! '-J in LA inir \ Mt vf ΙΛ LA IALA A

ffi! K la <f r-i r- ro * σ I A co l '»VD Γ * · σ' CO. · · · · · O · σ * “5 Mf st <j- <t -sf-

N

W

• file ® CO iH CM <f 00 <f I 1 p CM rH 00 r-t O 'ti * "' rrt A · · · · · · O S I-J CO <t <t ^

In peace OO

O <J- <t <f Mf CO cvi 9% i # * ·

CM Kl ro co CO CO CO

+ j i <u s

• ti i S o ro cm o cm '-J

«I r — I CM rsN CM CM <f + jj-H S ·« ro. · · · ΟίΓ «Ι? ° ^ ° ° ° <H I ly> Zi tih H! Λ / -s ^

ni S λ g £ £ S

(Dl · ^ ^ ^ got I {fl · al I X Sh viinvo r * oo σ>

Em I H f3. H r- (i-i · - <^ 147883 64

Table 1 continued

Ex. No. Microanalysis (%) Empirical formula found / calculated (solvate)

C Η N S

14 (b) 48.2 3.8 10.5 5.9 C H J SO.

48.2 3.6 10.7 6.1 ^ 15 (b) 50.1 4.3 12.4 7.2 C10H. "N.S07 49.8 4.2 12.9 7.4 18 18 4 7 16 (b) 49.5 3.65 9.6 5.8 C" _H pN SO? After 49.9 4.15 10.0 5.8 (fm ^ H '0) 17 (b) 50.6 5-4 10.5 C "nH" N.SOfl 51.8 5.2 10.8 1 2 3.22 4 8 LO.4 mol (i-Pr) 20] (b) 52.6 4.3 11.9 7.1 C1QHonN.S0 , 52.2 4.3 11.9 6.8,} 9.20 4 7. ,.

(0.2 mol of anisole) 2 (b) C19H21N4S2 ° 7Na 3

Footnotes (1) for the compound * D = _ £ _

PAC

R 1 of (6R, 7R) -3-acetoxymethyl-7-phenylacetamido-ceph-3-em-4-carboxylic acid Solvent system A is n-butanol / ethanol / water 4: 1: 5. Solvent system B is n-propanol / water 7: 3.

(2) inf. indicates an inflection.

(3) Trifluoroacetic acid spectrum.

Claims (2)

147683 1. Analogous process for the preparation of cephalosporin compounds of the general formula Η H s R1. C. CONH - \ 1 J-N J_CH2O.CO.NH2 I 'n' or 20 COOH where R4 represents a furyl group, thienyl group or phenyl group 2 R is an alkyl group having 1-4 carbon atoms, a cycloalkyl group having 3-7 carbon atoms, or a phenyl group which compound is a syn isomer or is present as a mixture of syn and anti isomers containing at least 90% of the syn isomer or non-toxic salts, biologically acceptable esters, 1-oxides or solvates thereof, characterized in that: a) condensing a compound of the general formula H2N1! III JN Λ-CHoO.C0.NHR12 stZ COOR where B represents> S or> S— ^ 0, where R 1 represents hydrogen or 12 a carboxyl blocking group, R represents hydrogen or an N-protecting group and where the broken line between the positions 2, 3 and 4 of the molecule indicate that the compound may be a ceph-2-em or ceph-3-em compound, or an acid addition salt or N-silyl derivative thereof, with an acylating agent corresponding to the acid R1.C.COOH k Iv Wherein R and R have the above meanings, or