DE1955475A1 - Cephalosporin compounds and their manufacturing process - Google Patents

Description

Dr. F. Zumstein «en. - Dr. E. Assmann Dr. R. Koenigsberger - Dipl. Phy3. R. Holzbauer

Dr. F. Zumstein jun.

Patent attorneys

8 Munich 2, Brauhausstrasse 4 / III

Case 92-604

Glaxo Laboratories Limited, Greenford, Middlesex, UK

Cephalosporin compounds and their manufacturing process.

The present invention relates to improved cephalosporin C analogs and their improved manufacture.

The cephalosporin compounds referred to in the present specification is referred to, v / earth is named very generally with reference to the cepham ring structure. The cepham ring is as follows shown:

(D "

(cf. J.A.C.S., 1962, £ j_ 3400 and J. Chem. Soc. 1965, 5031). The term "cephem" refers to the underlying Cepham ring structure with a single double bond.

In the manufacture of cephalosporin antibiotics and their intermediates it is often necessary to use intermediates such as 3-acetoxymethyl-73-aminoceph-3-em-4-carboxylic acid (commonly known as 7-aminocephalosporanic acid or 7-ACA) a sequence of

009819/1884

Submit transpositions to obtain the desired new connection. It may be undesirable to acylate the 7β-amino group early in the synthesis with the desired acylating agent. This may be the problem because conversions that are carried out anywhere in the molecule, for example nucleophilic displacement reactions at the 3-position, cause undesired conversions at the newly introduced acyl group in the 7ß-position. But letting the 7.beta.-amino group unprotected, it can be gen also subject to undesirable Ums3tzun _T. It has now been found that the 7β-amino group can be protected simply and economically by SOrmylation, while reactions are carried out elsewhere in the molecule and the 7β-pormamido group can then easily be broken down with regeneration of the 7β-amino group. In addition, the formamides do not normally appear to undergo racemization, as is the case with peptides of the Pail in similar systems. The formation of these 7β-pormamido compounds is therefore an extremely useful technique in cephalosporin chemistry.

According to the present invention, therefore, a method for Preparation of a 7β-aminocephem-4-carboxylic acid or a derivative thereof, which includes the following steps: Pormylation of a 7β-aminocephem-4-carboxylic acid or a derivative thereof with a group other than the desired group in the 1-, 2-, 3- or 4-position, with one or more reactions, which relate to the 1-, 2-, 3- or 4-position are carried out;

N-deformylation of the resulting compound to obtain the desired 7β-aminocephem-4-carboxylic acid or its derivative.

The method according to the invention can take place simultaneously Implementation that has more than one of the 1, 2, 3 or 4 positions relates to or involves a pole of two or more conversions affecting one or more of these positions.

009819/1884

The 7ß-Aminocephem-4-carboxylic acid or a derivative thereof with a other than the desired group in the 1-, 2-, 3- or 4-position, can be a Ceph-3-em or a Ceph-2-em compound.

The Pormylierung can be prepared using any suitable Fonnylierungsnittel3 such as formic Susanna with a Niedrigalkanearbonsäureanhydrid such as acetic anhydride, for example at a i'enperatur of up to 70 ⁰ C, preferably of 15, carried out to 40 ⁰ C. Alternatively, the formation can be carried out with a formic acid ester, for example by refluxing the ethyl formate with, if desired, a bifunctional catalyst such as pyrid-2-one or imidazole. Other alternative formylating agents are orthoformates, for example lower alkyl orthoformates, such as methyl or ethyl orthoformate. Other formylating agents include formyl halides, e.g., formyl fluoride or formyl chloride; or formic acid, zE at 100 ⁰ C a.

The step of introducing the desired group in the 1-, 2-, 3- or 4-position can be a.B. an oxidation reaction in the 1-position to be a sulfonyl or sulfoxide compound produce or more generally a nucleophilic displacement reaction at the 3 position. The present invention provides soir.it a convenient method, the 3-acetoxy group of 3-acetoxymethyl-7ß-aminoceph-3-en-4-carboxylic acid to displace nucleophilic. The present invention also provides a convenient method of substitution the '3-hydroxyethyl group of the 7ß-amino-3-hydroxyEie ~ ethyl-ceph-3-en-4-carconic acid.

Nucleophilic displacement reactions at the 3-Stellurg can after any of the usual methods can be used. So can for example, starting from 7-ACA, direct displacement of the 3-substituent-oen by reaction in a polar medium is old the appropriate nucleophilic reagent. Alternatively, one can do an indirect exchange up front by first using the 7-ACA or corresponding 3-hydroxy compound into a compound

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BW ORIGINAL

transferred, which has the group -GH ₂ X "in the 3-position, where X is a substituent which can easily be replaced by a liukleophil. The conditions for the nucleophilic displacement reactions are more detailed in British patents 912 541 and 1 012 943, in the Belgian patent 719 711 and in the German patent specification (German patent application P 19 30 204.6)

described.

Two important Verbijidungsreihen that can be obtained by nucleophilic displacement reaction at the 3-position, are the group-cephem-4-carboxylic acids with a 3-ether or 3-thioether f and their derivatives. Particularly important compounds of this series are those compounds in which the substituent in the 3-position is an I-lower alkylthiomethyl group.

The 4-carboxyl group can also be esterified, while the 7β-amino group is protected, using methods known in cephalosporin chemistry for carrying out such an esterification applies. The esterification can be carried out to give ester residues of any desired alcohol or phenol supply, e.g. of benzyl alcohols such as p-methoxybenzyl alcohol, Di-p-methoxyphenylmethanol, triphenylmethanol, diphenylmethanol, Benzoyloxymethanol or p-nitrobenzyl alcohol; Furfuryl alcohol; t-butanol or 2,2,2-trichloroethanol.

The N-deformylation of the 7β-pormamidocephem-4-carboxylic acid or its derivatives can be achieved by any suitable technique for the N-deacylation of an acylamido group. Thus, the acid-catalyzed hydrolysis or alcoholysis, the 7.beta.-iOrmamidogruppe with a Hineralsäure at a temperature of -15 ° to 100 ⁰ C, preferably +15 to 40 ⁰ C, is reached. A suitable reagent for the H-deformylation is concentrated chloro / aqueous acid in methanol or, preferably, in dioxane or tetrahydrofuran, since this avoids undesired transesterification reactions which can occur in methanol.

The 7ß-amino compound ₁ 2ε & νρ. «Εύ.φ « JUäIösIiches salt, e.g. as hydro- ^: "'"■■■' * "**" BAD ORIGINAL

chloride or hydrogen p-toluenesulfonate, or it can be precipitated by adjusting the pH value (e.g. to the isoelectric point), if necessary with extraction with a suitable solvent.

Certain 7ß-IOrmamidoverbindungen that in the invention Methods obtained are new and an important class of such compounds are those of the formula:

HCOHH

COOH

wherein R represents an alkyl group having 1 to 4 carbon atoms and X = 0-, -S-, -S- or §, and their salts and esters.

Il "" ή
0 0

The following examples explain the invention without, however, restricting it.

To the examples:

at
The NMR spectra were obtained / 60 MIz.

Paper electrophoresis was performed on Whatinan paper MM ITo. 3 at pH 1.9 [98% formic acid (16.7 ml), glacial acetic acid (84 ml), acetone (105 ml) and water (495 ml)] with a potential gradient of 25 volts / cm for 90 minutes.

The solvent systems used in the paper chromatographic investigations were as follows:

System 1: ethyl acetate: n-butanol: 0.1m sodium acetate (pH 5.0) = 8: 1: 8, descending with the upper phase (in equilibrium

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BATH

with the lower phase) as developing solvent, at 38 ⁰ O, on V / Hatman paper No. 1, buffered to pH 5.0.

System 2: n-propanol: water = 7: 3, descending at room temperature on Whatinan paper Ιίο. 1, buffered to pH 5 with 0.1 sodium acetate.

System 3: comprises the upper phase of the thin-layer chromatography below system described, in descending order, on Whatman paper ITo. 3 HM, buffered to pH 6 with 0.05 m-l sodium | dihydrogen phosphate.

Thin-layer chromatography (TLC) was performed using silica gel layers carried out with a solvent that is the upper phase of the system n-butanol: ethanol: water = 4: 1: 5. The chromatography papers and plates were coated with potassium jo & platinum sprayed and examined under UV light for absorbing and fluorescent stains. The presence of the 7ß-amino connections in the otromatograms can be done by spraying with liinhydrin as such compounds usually form a yellow color.

Petroleum ether is the fraction with the boiling range of 40 ^to) 60 °.

The reaction mixtures were dried over dried magnesium sulfate and evaporated at 40 C at approximately 20 mm Hg, unless otherwise stated.

Example 1 - '

Preparation of 7ß-amino-3-methylthiomethylceph-3-em-4-carboxylic acid.

The title compound was prepared via the following reaction scheme (the steps are described in more detail below),

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3-acetoxymethyl-7β-aminoceph-3-em-4-carboxylic acid

(a)

3-acetoxymethyl-7β-foi3namidoeeph-3-em ~ 4-carboxylic acid

Sodium 3-acetoxymethyl-7β-formamidocepli-3-em-4-carboxylate

(c)

7β-Formamido-3-methylthiomethylceph-3-eni-4-carboxylic acid

(d)

7ß-Amino-3-methylthiomethylceph-3-em-4-carl> onic acid,

a) 3-Acetoxymethyl-7ß-forraamidoceph-3-em-4-carboxylic acid.

3-Acetoxymethyl-7ß-aminoceph-3-em-4-carboxylic acid (50.0 g) was dissolved in acetic anhydride (75 ml) and formic acid (400 nil) at 20 ^° C. with stirring. When complete solution was achieved (approximately 5 minutes) the solvents were evaporated (rotary evaporator, oil pump) leaving a resin which was shaken with ethyl acetate (250 ml). The insoluble material was removed by filtration and the piltrate was evaporated to dryness, leaving a resin which was triturated under ether and petroleum ether (b.p. 40-60 °) and 3-acetoxyraethyl-7β-fonaamidoceph-3-em-4-carboxylic acid as a light yellow powder (43.6 g), DSC: R _p = 0.19, O] ^ ⁰ + 113.5 ° (c = 1.00, dioxane), λ max (pH6 buffer) 259 µm. (<f = 8400); V ^ iaax (Nujol): 1772 (ß-Lactaia) 1748 (acetate), 1708 (carboxylic acid), 1654, 1548 cm "* ^1. (Amide) j" T (D ₂ O with UaHCO ₃ ): 1.67 ( 0.9 proton singlet: CHO) ₁ 1.75 (0.1 proton singlet: CHO), 4.20 (1 proton doublet, J - 4.5 Hz .: C-7 H), 4.80 (1 proton

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BATH ORIGINAL

Doublet, J = 4.5 Hz: C-6 H), 5.00, 5.26 [two 1-proton doublets (branches of a quartet), J = 13 Hz: C-3 CH ₂ ], 6.24 , 6.64 [two 1-proton doublets (branches of a quartet), J = 18 Hz .: C-2 H ₂ ] 7.86 (3-proton singlet: CH ₅ CO) (it can be seen that eB "ice" and " trans "forms in the secondary amide part).

b) Sodium ^ -acetoxymethyl ^ ß-foimamidoceph ^ -em ^ -carboxylate.

10 g of 3-acetoxymethyl7β-formamidoceph-3-em-4-carboxylic acid were stirred in acetone (120 ml) for 10 minutes, the mixture was filtered to remove traces of the plague substance and the piltrate was washed with a 10bigen (w / v. ) Solution of sodium 2-ethylhexanoate in acetone (60 ml). After cooling for one hour, the precipitate was isolated by filtration, washed with acetone and dried in vacuo, giving sodium 3-acetoxymethyl-7β-formamidoceph-3-em-4-carboxylate as a gray-white test substance (10.0 g), [oc] ^ ° + 121.3 ° (c = 1.00, water), λ _mQY (V / water) 260 mn (£ = 8,600); V _n , .- (Nujol): 1760 (β-lactarn), 1730 (acetate), 1672, 1544 (amide), 1600 cm "" ¹

c) 7β-IOrmamido-3-methylthiomethylceph-3-a-4-carboxylic acid.

Sodium 3-acetoxymethyl-7β-formamidoceph-3-em-4-carboxylate (3.22 g, 0.01 mol) in Yfasser (50 ml) was treated with methanethiol (1.44 g, 0.03 mol) in a sealed glass tube at 70 ° for 2 hours. The contents of the tube were placed in an open vessel transferred and stirred for 15 minutes at room temperature to allow the excess thiol to evaporate. The solution was covered with ethyl acetate (50 ml) and the pH of the aqueous layer adjusted to 1. The ethyl acetate layer was isolated and washed with further ethyl acetate extracts (2 × 50 ml) of the aqueous layer united. The combined extracts were washed successively with water (50 ml) and brine (50 ml), dried, the solution was evaporated, leaving an impure 7β-JEOrmamido-3-methylthiomethylceph-3-em-4-carboxylic acid as a colorless foam (2.22 g) remained, which was dissolved in 12 ml of acetone and 12 ml of ether and a 1C (w / v) solution of sodium 2-ethylhe was given xanoate in 12 ml of acetone. After cooling for an hour

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the precipitate was isolated by filtration, washed with acetone: ether (1: 1, 2x10 ml) and ether (2 × 20 ml) and dried in vacuo, using sodium 7β-formamido3-methyItMomethylceph-3-em-4-carboxylate obtained as a colorless plague substance (1.69 g), which was found to be practically homogeneous in thin-layer chromatography (a trace of the starting material, cephalosporanic acid), O] p ° + 75.5 ° (c = 1.00, V / water), λ _max (pH 6 buffer) 263 nm (£ = 7760); Y _mav (Nujol): 1760 (β-lactam), 1680, 1540 (amide), 1608 cm "'(GO ₂ "); Ί, 78 (1-proton singlet, CHO), 4.31 (1-proton doublet, J = 4.5 Hz .: C-7H), 4.84 (1-proton doublet, J = 4.5 Hz. : C-6H) 6.20, 6.65 [two 1-proton doublets (branches of a quartet) J = 17 Hz .: C-2 H ₂ ], 6.22, 6.69 [two 1-proton doublets ( Branches of a quartet), J = 14 Hz .: 0-3 CH ₂ ], 7.98 (3-proton singlet; SCH,). *) Τ (D ₂ O):

For the title compound, the following in-yitro biological results were obtained obtain:

Staph. aureus 604 ^ 2.5; Staph. aureus 663 2.5; Staph aureus 3452 4; Staph. aureus 11127 4; E. coli 573 52; S.

at 804 62; and Pr. mirabilis 431 [all by dilution test in tube (Jf / ml)].

Protection of the mouse (ED ™ mg / kg / dose) against Staph. aureus 11127 (administered by subcutaneous injection) was 30. 4.7 # of the compound were extracted from the urine of female rats after oral Administration won.

7ß-Amino-3-methylthiomethylceph-3-em-4-carboxylic acid and its hydrochloride.

d) (i) 7β-Formamido-3-methylthiomethylceph-3-em-4-carboxylic acid (2.88 g) was dissolved in methanol (25 ml) and the solution was stirred with concentrated hydrochloric acid (4.0 ml) treated. After 1.5 hours at room temperature, the solution was diluted with 80 ml of water und.mit ethyl acetate (2 × 30 ml) and Ether (2 × 20 ml) washed, evaporated on a rotary evaporator, to remove the remaining ether and adjust the pH with

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- ίο -

0.88On ammonia adjusted to 3.5. The precipitate was isolated after standing overnight in the refrigerator, washed with water (2 × 10 ml), methanol (2 × 10 ml) and ether 3 × 20 ml), then dried and gave 7β-amino-3- wethylthIomethylceph-3-em-4-carboxylic acid as a colorless powder (1.41 g), Ep. = 222 to 225 ° (decomposition), R _f = 0.60 (system 2), DSC: R _f = 0.15, in electrophoresis at pH 1.9 "the substance migrates 2.0 cm towards the cathode (3-acetoxymethyl-7ß-aminoceph-3-em-4-carboxylic acid migrates 1.85 cm under identical conditions), λ ___ (pH6- Buffer):

266 mn (£ = 9590); V _mQ ^ (ffujol): 1804 (ß-lactam) 1536 cm " ¹ (carboxylate)," f ( ^D 2 ° ^{with NaHCO} 3) ^: 4 »53 ^ 0.5 proton doublet, J = 4.5 Hz. :; 0-7 H), 4.88 (1-proton doublet, J = 4.5 Hz .: C-6 H), 5.21 (/ V 0.5-proton doublet, J = 4.5 Hz .; C-7 H), 6.14, 6.63 [two 1-proton doublets (branches of a quartet), J = 18 Hz »; C-2 H ₂ ], 6.21, 6.68 [two 1-proton doublets (branches of a quartet), J = 14 Hz .; C-3 CH ₂ ], 7.95 (3-proton singlet; SCH ₃ )

Analysis: C. ι
■ 9 ^H 1 ₂ N ₂ O ₃ S (260) 4, 6 * N 1 ο, 8th Calculated: C. 41 , 5 ί 5 H. 4, 6th 1 ο, 2 Found: 41 , 2

S 24.6 <fo 23.6

(The signal in the weak peId for the 7-proton v / is on a Carbonation of the 7ß-amino group).

d) (ii) A solution of 7β-i "onnamido-3-methylthiomethylceph-3-em-4-carboxylic acid (6.4 g) in 50 ml of dioxane was treated with 50% aqueous hydrochloric acid (7 ml) The mixture was lyophilized for 2 hours at room temperature. The resulting material was washed with ethyl acetate and ether and dried to give 7β-amino-3-methylthiomethylceph-3-em-4-carboxylic acid hydrochloride (5.95 g, $ 90) as white> amorphous powder substance, [aj-p + 58.4 ° (c = 1.3 $, BaHCO ₅ ); y \ (0.1 M pH 6 phosphate buffer) 265, Snm ( 6 - 7650); V «" ^ (Nunöl):

* λ ' max

1782 (β-lactam). 1715 and 1682 (carboxyl) cm "¹; X (D ₂ O NaHCO ₅ ): 6.59 ^ / 1-proton doublet, J = 4.5 Hz), 6.91 (C-6 H 1-proton doublet , J = 4.5 Hz), 6.2 and 6.67 [C-2 H ₂ two 1-proton doublets (branches of a quartet), J = 18 Hz], 6.24 and 6.7 # C-7H

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[C-3 CH «two 1-proton doublets (branches of a quartet), δ = 13 Hz], and 7.98 (3-proton singlets, CH ₅ S).

Example 2

■ ■ ■ ■ ■ »^ Wl ■■■■ —— ■■

a) 3- (3-Carbamoylpyridiniummethyl) -7β-formamidoceph-3-eiit-4-carboxylate.

A solution of 3-acetoxymethyl-7β-formamidoceph-3-em-4-carboxylic acid (20 g, 0.06 mol) and nicotinamide (29.3 g, 0.24 mol) in water (300 ml) was added for 15 hours held at 50 ° for a long time. The cooled reaction mixture was filtered and the piltrate was adsorbed on a 60 cm column of a "Deacidite FF" ion exchange resin. The column was eluted with distilled water and 100 ml fractions were collected. The optically active fractions were combined and subjected to freeze-drying, the solid substance obtained was slurried with acetone (3 × 100 ml) and gave the betaine sought as a white powder (7.1 g, 30 Jt), BpjKJA ⁼ ° » ¹ ^ (system 2), R ^ = 0.11 (electrophoresis, pH 1.9, 500 V, 2 hours), [α] _β -5 ° (c = 1.00, water), λ 262 nm (£ = 10 300) ; V _ffieY (liujol): 1770 cm " ¹ (β-lactam), 1675 and 1528 cm" ¹ (Aoid), and 1610 cm " ¹ (CO ₂ "); Z (D ₂ O): 0.31 to 2.0 (4-proton complex; pyridinium), 1.74 (0.9-proton singlet; CHO), 1.88 (0.1-proton singlet; CHO), 4.77 and 4.20 (two 1-J proton doublets, J = 4.5 Hz .; 6- and 7-H), 4.38 and 4.50 [two 1-proton doublets (branches of a quartet, J = 14 Hz, 3-CH ₂ )], 6.36 and 6.79 [two 1-proton doublets (branches of a quartet), J = 18 Hz., 2-H].

Analysis; C j ^ H ^ 4 ^H 45 Calculated: C 49.7 % H 3.9 £ N 15.5 $ S 8.8 %

Found: 48.3 4.5 15.7 6.9

b) 7β-Amino-3- (3-carbamoylpyridiniummethyl) -ceph-3-em-4-carboxylate-bis-hydrochloride.

3- (3-Carbamoylpyridiniummethyl) -7ß-formamidoceph-3-em-4-carboxy-

009819/188 / »

Lat (2 g) was added in part to a mixture of concentrated hydrochloric acid (3 ml) and methanol (40 ml) with stirring to give an orange solution which was stirred at room temperature for 4 hours. The reaction mixture was diluted with ethyl acetate (150 ml) and the precipitated pest substance was isolated by filtration, whereby the title compound was obtained as a gray-white powder (1.13 g, 48 %) , ¹¹ PACA ⁼ ° ^'09 ( ^s y ^{stem 2} ) » ^R p = ° 14 (electrophoresis, pH 1.9, 500 V, 90 minutes), [a] _D -65 ° (c = 1.00, water), λ _max 263 nm (ξ = 8400 ); Y_ '(Hujol): 1780 cm " ¹ (β-lactam), 1687 (COIiH), and 2600 cm" ¹ (NH ₅ ⁺ ), 1687 cm " ¹ (-CO ₂ H). T (D ₂ O): 0.3-1.8 W (pyridinium protons), 4.02 and 4.48 [two i-proton doublets (branches of a quartet)., J = 18 Hz, 3-CH ₂ ], 6.11 and 6.56 [ two 1-proton doublets (branches of a quartet), J = 17 Hz., 2-H], and 4.54 and 4.69 (two 1-proton doublets, J = 4.5 Hz., 6- and 7 -H).

Analysis; C ₁ ^ H ₁ gN ^ SO ^ Cl ₂

Calculated: C 41.2 # H 3.9 f N 13.9 1 » S 7.9 1 ° Cl 17.5 1 <> Found: 36.7 4.3 13.5 6.2 19.6

Example 3

a) 3- (4-Carbamoylpyridiniummethyl) -7β-formamidoceph-3-em-4-carboxylate.

3-Acetoxymethyl-7β-formamidoceph-3-em-4-carboxylic acid (20.0 g) and isonicotinamide (29.3 g) were dissolved in 300 ml of water and kept at 50 ° for 15 hours. The cooled reaction mixture was filtered, passed through a Deacidite IT ion exchange resin column (60 cm) and the column was eluted with distilled water. The combined optically active fractions were subjected to freeze-drying and the resulting powdery substance was slurried with acetone (3 × 100 ml), the title compound being obtained as a white powder (7.2 g, $ 25), Rp _ACA = 0.18 (system 2 ), R _F = 0.15 (electrophoresis,

009819/1884

pH 1.9, 500 T, 2 hours), [α] ^ -5 ° (c = 1.00, water), Λ 262 mn (2 = 13ΟΟΟ); V " _ma " (Nujol): 1770 cm " ¹ (β-lactam), 1675 and 1528 cm" ¹ (COiIH), and 1610 cm " ¹ (CO ₂ "); TT (D ₂ O): 1.73 (0.9 proton singlet, CHO), 1.88 (0.1 proton S / .nglet, CHO), • 0.82 to 1.57 (4 proton complex ; Pyridinium), 4.19 and 4.75 (two 1-proton doublets, J = 4.5 Hz., 6- and 7-H), 4.26 and 4.58 [two 1-proton doublets (branches of a Quartet), J = 14 Hz., 3-CH ₂ ], 6.25 and 6.79 [two 1-proton doublets (branches of a quartet), J = 18 Hz., 2-H].

Analysis: C 15th 14 4 SO ₅ • 2H ₂ ⁰ 6th N 14, 1 ok S. 8.1 Calculated: C. 45, 2 io - H '4, 2 14, 9 7.1 Found: 44, 1 4,

b) 7β-Amino-3- (4-carbamoylpyridiniummethyl) -ceph-3-em-4-carboxylate-bis-hydrochloride.

3- (4-Carbamoylpyridiniummethyl) -7β-formamidoceph-3-em-4-carboxylate (1 g) was added to a mixture of 20 ml of methanol and 2 ml

concentrated hydrochloric acid to give a light yellow solution. After 10 minutes a white solid precipitated and the suspension was stirred for 4 hours at room temperature. The solid was isolated by filtration to give the title compound as a white powder (600 mg). [A second, less pure fraction was isolated by diluting the filtrate with ethyl acetate], R ^ = 0.4 (electrophoresis, pH 1.9, 500 V, 90 minutes), ^ _kQk = 0.08 (system 2), [ a] ^ - 35 ° (c = 1.00, water), Λ _ηΆ * 263 nm ( £ = 9400); ψ _ηαΎ (Nujol): 1780 cm " ¹ (ß-lactam), 1687 cm" ¹ (CONIl), 1687 (CO ₂ H) and 2600 cm "" ¹ . (NH ₅ ⁺ ); X (D ₂ O): 0.74 to 1.48 (pyridinium protons), 4.04, 4.47 [two 1-proton doublets (branches of a quartet), J = 14 Hz., 3-CH ₂ ], 4 , 54 and 4.69 [two 1-proton doublets, J = 4.5 Hz., 6- and 7-H], 6.11 and 6.56 [two 1-proton doublets (branches of a quartet), J = 17 Hz., 2-H] ..

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- H-

Analysis; C ₁₄ H ₁₆ H ₄ SO ₄ Cl ₂

Calculated: C 4-1.3 # H 3 ^.; # S 7.9 # Cl 17.5 $ N 13.9 # Found: 37.2 4 6.2 19.6 13.5

Example 4

a) Diphenylmethyl-7ß-formf ^: .vio-3-methylthiomethylceph-3-em-4-carboxylate.

A solution of sodium 7β-formamido-3-nietliylthiomethylceph-3-em-4-carboxylate (6.20 g, 0.02 mol) in v / water (50 ml) was covered with 50 ml of ethyl acetate and the pH to 2 , 5 adjusted (2N hydrochloric acid). The ethyl acetate layer was isolated, combined with further ethyl acetate extracts (2 × 50 ml), washed with 50 ml of water and 50 ml of brine, dried (magnesium sulfate), the solvent evaporated, leaving a foam which was dissolved in tetrahydrofuran (250 ml) and with diphenydiazomethane (3.88 g, 0.02 mol) in petroleum ether (b.p. 60 to 80 °; 70 ml) was treated. The solution was kept in the dark overnight, a small amount of glacial acetic acid was added to destroy unreacted diphenyldiazomethane and the volume of the solution was reduced to 30 ml, then diluted with 200 ml of ethyl acetate. This diluted solution was washed with 3% sodium bicarbonate (5 × 100 ml), water (2 × 100 ml) and brine (100 ml), dried (magnesium sulfate), and the solvent was evaporated to give an orange-colored oil (7.22 g) remained, TLC (benzene: ethyl acetate = 2: 1 as developing solvent), R ₁ , = 0.62, 0.80 (trace), 0.95. This oil was dissolved in benzene (80 ml), then petroleum ether (b.p. 40 to 60 °; 250 ml) was added, a resin which was triturated under petroleum ether and diphenylmethyl-7ßformamido-3-methylthiomethylceph-3- yielded em-4-carboxylate as a brown ¹ powder (3.02 g). DSC: Rj = 0.62, = 60 to 65 ° Fp, Ea] ₇₁ -24.5 ° (c = 1.00, dioxane), λ _MQV (ethanol) 268 nm (£ =.

J- * 1- * IUcLa.

7000); V " _max (CHBr ₃ ) *. 1780 (β-lactam), 1712 (ester), 1690, 1498, cm" ¹ (COITH), Z (CPGl ₅ ) 1.68 (1-protou singlet; 'CHO ), 2.58 (1-proton singlet; aromatic protons), 2.93 (1-proton singlet, Ph ₂ CH), 3.32 (1-proton double, J = 9 Hz .; CONH), 4.08 (1-proton double doublet, J- = 5.9 Hz; C-7 H),

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SAD

4.92 (1-proton doublet, J = 5 Hz .; C-6 H), 6.29, 6.56 [two 1-proton doublets (branches of a quartet), J = 14 Hz .; C-3 CH ₂ If 6.42 v.2 proton singlet; C-2 H ₂ ), 8.11 (3-protome <singlet j CH ₃ S).

Analysis: C ₂₅ H ₂₂ H ₃ O ₄ S ₂ (454.6)

Calculated: 0 60.9 1 ° H 4.9 # II 6.2 i » S 14.1 ° /> found: 61.6 5.3 5.5 13.5

b) Diphenylmethyl-7β-amino-3-niethylthioniethylceph-3-a-4-carboxylate.

Diphenylmethyl ^ ß-forraamido ^ -nethylthiomethylceph ^ -emH-carboxylate (0.88 g, 0.0019 mol) in 10 ml of methanol vmrde drop-white treated with concentrated hydrochloric acid (1.0 ml). A colorless precipitate separated out immediately, but this dissolved after stirring for 5 minutes. After 15 minutes, however a second precipitate precipitated out and after the mixture had been cooled for 2 hours the solid came through Isolated by filtration, washed with a little water and then dried over phosphorus pentoxide. The dried plague substance was stirred with ethyl acetate (15 ml, 2 hours, χ 2) in order to leach out unchanged starting material, the remaining solid substance was isolated by filtration, washed with ethyl ether, dried and gave diphenylmethyl-7β-amino-3-methylthiomethylceph ^ -em ^ -carboxylate hydrochloride (0.35 g), DSC Rp = 0.83.

Analysis: ^G 22 ^H 23 ^1T 2 ⁰ 3 ^S 2 ^C1 ( ⁴⁶⁵ »°)
Calculated: Cl 7.7 ^C A
Found: Cl (ionic) 7.6 Ji

A portion (0.33 g) of this hydrochloride was dissolved in a mixture of 3 strength sodium bicarbonate (30 ml) and ethyl acetate (30 ml) stirred until dissolved (2 hours). The ethyl acetate layer was isolated and further ethyl acetate extracts (25 ml)

009819/188 /.

of the aqueous layer combined, the combined Ex!. -3-em-4-carboxylate (0.28 g), obtained, DSC: R ™ = 0.83, pp. = 119 to 123 °, [Ot] ₃₅ -81.4 ° (c = 1.00, Dioxane), J _{m & x}

(Ethanol) 270 mn (S = 7130), f _(CHB} ^ ₉₀ ^ _{20 {m)}

λ max yc

_(CHB} ^ ₉₀ ^ _{20 {m} λ max y c.

1768 (ß-lactam), 1712 cm " ¹ (ester); % (CDCl ₅ ): 2.86 (10-proton singlet; aromatic protons), 3.05 (1-proton singlet; Ph ₂ CH), 5, 03 (1-proton doublet, J = 5 Hz .; C-6 H), 5.29 (1-proton ₇ broad doublet, J = 5 Hz; C-7 H), 6.37, 6.67 [ two 1-proton doublets (branches of a quartet), J = 13.5 Hz .; 0-3 clip], 6.45 (2-proton singlet; C-2 H ₂ ), 8.16 (3-proton singlet; SCH ₅ ), 8.22 (2-proton-wide singlet; NH ₂ ). A portion (0.200 g, 0.0005 mol) of this amine was dissolved in 4 ml of ethyl acetate and 2 ml of ether and a solution of p-toluenesulfonic acid monohydrate '(0.089 g, 0.0005 mol) was added. The precipitate was isolated by filtration, washed with ethyl acetate: ether = 1: 1, dried to give diphenylmethyl-7β-amino-3-methylthiomethyl ceppli-3-em-4-carboxylate hydrogen p-toluenesulfonate as a colorless powder (0.126 g), pp. = 159 to 161 ° (decomposition), [a] _D -33.2 (.c = 1.00, dimethylacetamide), λ _max (ethanol) 264 nm (€ = 7560); fmax (CHBr ₅ ): 2600 (HH ₃ ), 1785 (β-lactam), 1720 (ester), 1005 cm "* (SO ₅ "); T (CDCl ₅ with DMSO d-6): 1.6 to 2.3 (3-proton singlet; NH ₅ ), 2.23, 2.90 [two proton doublets (branches of a quartet), J = 9 Hz. ; p-disubstituted aromatic protons], 2.70 (10-proton singlet; aromatic protons), 3.11 (1-proton singlet; Ph ₂ CH), 4.88, 4.98 (two 1-proton doublets, J = 5 Hz; C-6 H, C-7 H), 6.14, 6.47 [two 1-proton doublets (branches of a quartet), J = 13.5 Hz .; C-3 CH ₂ ], 6.30, 6.85 [two 1-proton doublets (branches of a quartet), J = 16 Hz .; C-2 Hg], 7.68 (3-proton singlets; CH-C-), 8.16 (3-proton singlets; CH ₅ S).

Analysis: ^C 29 ^H 30 ^N 2 ° 6 ^S 3 (598.8)

Calculated: C 58.5 5 $ H 5.0 # N 4.7 Io S 16.0 # Found: 58.7 5.0 4.3 15.6

9 8 1 9/1 8 8 4

Example 5

Diphenylmethyl ^ ß-amino ^ -methylthiomethylceph ^ -enW-carboxylate hydrochloride.

* A solution of 7β-i ¹ ormamido-3-methylthiomethylceph-3-em-4-carboxylic acid (4.8 g, 16.7 mmol) in peroxide-free dioxane (50 ml) was added to a solution of diphenydiazomethane (from 8.4 g benzophenone hydrazone: 43 mmol) in 150 ml ether. The solution was left to stand overnight at room temperature, then treated with glacial acetic acid (2.5 ml) and evaporated to give a light yellow oil which, according to the thin-layer chromatogram, consisted of two main components. The oil was taken up in 100 ml of ether, washed with saturated sodium hydrogen carbonate solution (50 ml), 2N hydrochloric acid (50 ml) and saturated sodium chloride solution (50 ml). The solution was dried (sodium sulphate) and evaporated to give an oil which directly was used for the deformylation attempt. The thin-layer chromatogram (benzene: ethyl acetate = 2: 1, on Merck ¹ s silica) showed two main components, which are a decomposition product of excess dipheny / diazomethane and diphenylmethyl-7β-formamido-3-methylthio * - methyleeph-3 ** eni ~ 4 -carbo: xylate corresponded.

The oil was digested in a mixture of peroxide-free tetrahydrofuran (120 ml) and concentrated hydrochloric acid (30 ml) and stirred at room temperature until the thin-layer chromatograph showed that no diphenylmsthyl-7ß-formamido * '3 "-methylthiomethylceph * -3-em «-4-carboxylate was more present» This was after about 5 hours. During this time the hydrochloride salt crystallized. The solution was placed in the refrigerator overnight, filtered, the solid substance dried, and diphenylmethyl-7ß was obtained -amino.-3-methylthiomethyloeph »3 ' ⁱ -em-4 ⁱ -oa £ boxylat-bydrochlorid (2.4 g), I * D. * 185 to 192 ° (decomposition), \ (EtOH) 265 im (j £ s 6850) JC «3 _D -24.9 ° (c« 1 * 0, BMSO) j \> * 2620 (-11H ₅ ), 1770 (ß-lactam), 1711 (ester), and 702 ca- * ayl); T (DMSö-d ₆ ): 8.1 (3-2rotOMiiSiagttlet1;} -s-CS »), 6.25 (2-roton singlet; C-2 protons), 6.05» 6.35 (two 1-iroto-

009819/1884

BATH ORIGINAL

1955A75

nen doublets, J = 18 Hz .; 3-CH ₂ -S), 4.70 ( ¹ OtOn doublet,

J = 5 Hz .; 7-H), 4.5 (1-proton doublet, J _; Iz .; 6-H), 2.9

(1-proton singlet; CHPh ₂ ), 2.45 (10-proton hultiplet; aromatic protons).

Analysis: ^C 22 ^H 23 Olli 2 ° 3 ^S 2 5 , U / O Cl 7, 7th Ii 6, O S. 1 3, 8th Calculated : C 57 , 2 $ H 5 , 0 7, 9 5, 8th 1 3, 1 Found: 57 , 3

Concentration of the mother liquor of the DeformyIierungsversuches and trituration with ether gave a further fraction of the hydrochloride k (2.1 g), which with the material obtained as described above; · Was identical. The total yield of diphenylmethyl-7β-amino-3-methylthiomethylceph-3-em-4-carboxylate hydrochloride was 4.5 g (60 %, based on 7β-pormaiaiclo-3-ethylthiomethylceph-3-em-4-carboxylic acid). The same compound was obtained in a yield of 38% if the deformylation was carried out in Dioxo.n, which contained 6N-hydrochloric acid.

Example 6

a) 3-Acetoxymethyl-7ß-formamidoceph-2-em-4cc-carboxylic acid.

3-acetoxymethyl-7.beta.-forinainidoceph-3-em-4 - carboxylic acid (3.0 g, 1 mmol) was suspended in 12 ml of dry pyridine at Bäumtemperatur). 1.2 oil of acetic anhydride was added to the suspension with stirring and the reaction was allowed to proceed for one hour * The pyridine salt crystallized on cooling with ice, it was filtered off the organic phase with water, drying and evaporation was obtained as a resin. The resin (0.61 g, 20.3 %) could not crystallize. The spectroscopic properties of the product agree with the structure of the compound mentioned in the title: Ψ (film) 3300 (BH), 1770 (3-lactam), 1730 (OGOGH ₃ ), 1675 (HCOM-); T (D ₂ O ₁ ZHaHGO ₃ ): 1.82 (singlet, HCO-K 3.52 (singlet, -aCH-S), 4.51 and 4.71 (double doublet, J - 4 Hz .; 6H and

-.'-..'-. * "0 0-98 18/188 4

BATH ORIGINAL

YH protons), 5.22 (collapsed quartet), 5.82 (singlet, CHCO ₂ H) and 7.89 (singlet, CH ₃ CpO-).

b) 3-Acetoxyinethyl-7ß-arainocepli-2-eEi-4a-carbonic acid.

3-acetoxymethyl-7 ^r <-fomaraidoccpk-2-on / yes ~ carboncäiire (1.5 g, .0.5 mmoles) was added in 12 ml dioxane, the 50 # Kona. Containing hydrochloric acid (1> 7 ml), dissolved at room temperature. After 2 hours, the excess solvent was removed by freeze-drying, giving the solid hydrochloride (1.5 g, $ 100). When the pH was adjusted to the isoelectric point with a dilute ammonia solution, no precipitation of the zwitterion was obtained. The IR and ITl-IR results with the solution from the freeze-drying agreed with the structure of the compound named in the title. Paper electrophoresis was performed on Whatman paper No. 1 at pH 1.9 [90% formic acid (16.7 ml), glacial acetic acid (84 ml), acetone (105 ml) and water (495 ml)] at a potential gradient of 125 volts / cm for 20 minutes. The resulting mobility was 5 cm in comparison with 3-acetoxyethyl-7i? -Aminoceph-3-em-4-carboxylic acid, which had a mobility of 3.5 cm.

c) Sodium 3-aceto> ^ methyl-7ß- (2 _t 6-dichlorobenzamido) -ceph-2-em-4a-carboxylate.

The hydrochloride of 3-acetoxynethyl-7ß-aminoceph-2-em-4a-carboxylic acid (1.5 g, 0.05 tall oil) was dissolved in refluxing dry chloroform (20 ml) containing 3.2 ml (0.015 liol) Hexamethyldisilazane, heated for 1 hour. The excess silylating reagent and solvent were removed under high vacuum. The resulting resin was redissolved in dry ethyl acetate (20 ml) and 2,6-dichlorobenzoyl chloride (1.0 6, 0.45 ml) was added. The reaction was carried out under reflux for 1 hour. Treatment of the organic phase with dilute hydrochloric acid, water, drying and evaporation gave a resin. When adding a solution of 10 μS ITatriuin-2-ethylhexanoate in ethyl acetate, a sodium salt precipitated out, ψ _ _v (HuJöl) 3300 (HH), 1760 (ß-lactam), 1720 (acetate), 1660 (amide) and

009819/1884 BAD ORfGlWAl

1955-75

1605 cm (-CO ₂ Na), Rj, (System 3) 0.53 (with one impurity, Rp = 0.19). These properties are compatible with the structure of the compound mentioned in the title.

0098 19/188 4

BATH

Claims (23)

- 21 claims 1. Process for the preparation of a 7β-aminocephein-4-carboxylic acid 'or a derivative thereof, characterized in that a 7ß-Aminocephem-4-carboxylic acid or one of its derivatives with a group other than the desired group in the 1-, 2-, 3- or 4-position, is formylated, one or more reactions be carried out at the 1-, 2-, 3- or 4-position, and the compound obtained is N-deformylated to give the desired To obtain 7ß-aminocephem-4-carboxylic acid or one of its derivatives. 2. The method according to claim 1, characterized in that the lOrmylation with formic acid together with a medrigalkanecarboxylic anhydride is carried out. 3. The method according to claim 2, characterized in that the lOrmylierung "is performed at up to 70 ⁰ C. 4. The method according to claim 3, characterized in that the 3? Ormy; 3? Ormylation at a temperature of 15 to 40 ⁰ C carried out 5. The method according to any one of claims 2 to 4, characterized in that that the IT-lower alkanecarboxylic anhydride is acetic anhydride. 6. The method according to claim 1, characterized in that the Formation with a formic acid ester is carried out. 7. The method according to claim 6, characterized in that the 3? Ormylation in refluxing ethyl formate is carried out. 8. The method according to claim 1, characterized in that the pormylation is carried out with an orthoformate. 0 0 9 8 19/188 A 9. The method according to claim 1, "characterized in that the PorrnylieruT-g is carried out with formic acid. 10. The method according to claim 1, characterized in that the formylation is carried out with an ITormyl halide. 11. The method according to any one of the preceding claims, characterized characterized in that the reaction to introduce the desired group is a nucleophilic displacement reaction at the 3-5 position is. 12. The method according to any one of the preceding claims, characterized in W, that the reaction is an esterification of the 4-carboxyl group to introduce the desired group. 13. The method according to any one of the preceding claims 4r characterized in that the IT defornylation catalyzed by acid Hydrolysis or alcoholysis "is effected. 14. The method according to claim 13, characterized in that the IT deformylation with a mineral acid at a temperature of -15 ° "to 100 ⁰ C is catalyzed. 15. The method according to claim 14, characterized in that the Nf deformylien to be led. N-deformylation at a temperature of + 15 ° Ms 40 ⁰ G through- 16. The method according to any one of claims 13 to 15, characterized in that the IT deformylation is carried out using hydrochloric acid in methanol, dioxane or tetrahydrofuran. ^; 17. The method according to any one of the preceding claims, characterized characterized in that the desired 7β-aminocephem-4-carboxylic acid or one of its derivatives is separated off as an insoluble salt or is precipitated by adjusting the pH. 0 0 9819 / 188Λ. . BATH ORIGINAL 18. The method according to claim 17, characterized in that the insoluble salt a hydrochloride or a hydrogen p-toluenesulfonate is. 19. The method according to any one of the preceding claims, characterized characterized in that the 7ß-Aminoeephen-4-carboxylic acid or one their derivatives with a group other than the desired group in the 1-, 2-, 3- or 4-position form a Cepli-3-em compound is. 20. Verfaliren according to one of claims 1 "to 17, characterized in that" that the 7ß ~ aininocephen-4-carboxylic acid or a derivative of which with a group other than the desired group in the 1-, 2-, 3- or 4-position is a Ceph-2-em compound. 21. Compounds of the general formula: Hcorai- I I (ID CH ₂ ZR COOH wherein R represents an Allry group with 1 to 4 carbon atoms and Z is -0-, -S-, -S- or _n_ and their salts and Ester. - 0 0 22. 7β-Foraaniäo-3-methylthiomethylceph-3-em-4-carboxylic acid. 23. Diphenylnetllyl-7β-formanido-3-methyllylthiomethylceph-3-en-4-carboxylate. Π09819 / 188 / BAD OBLOLHAt