Process for preparing new derivatives of cephalosporine

The invention relates to a process for the preparation of novel cephalosporin derivatives.

British Patent No. 1,399,086 discloses a number of cephalosporin derivatives of the general formula

<img file="CS264257B2_D0001.tif" />

CS 264 257 B2 where

R represents a hydrogen atom or an organic radical,

R^and means monovalent. an organic group bonded to an oxygen atom, a carbon atom and an ether forming group,

В means group \

s /

or \

S - * · O, and /

P is an organic residue.

However, R is not a 2-amino-4-thiazolyl group, nor is there any example in which P is a quaternary methylammonium group of a similar type to the compounds of Formula I below. U.S. Patent No. 3,971,778 and Excluded Patent Nos. 4,024,133, 4,024,137, 4,064,346, 4,033,950, 4,079,178,

091 209, 4,092,477 and 4,093,803 disclose similar compounds.

U.S. Pat. No. 4,278,793 describes a wide variety of cephalosporin derivatives of the general formula

<img file="CS264257B2_D0002.tif" />

where

R 1, R 2, R 3, R 4, X and A contain the meanings that correspond to the substituents in formula (I).

However, the 20 columns of the meanings of the various substituents on the 78 pages of the tables of the structural formulas and none of the 225 examples show an example in which A is a quaternary methylammonium group as contained in the compounds of formula I. British Patent Specification 1,604,971 a similar procedure is described. In British Patent Specification No. 2,028,305 A, which is related to the above specification, only A is present in the meaning of A.

U.S. Pat. No. 4,278,671 describes a process for the preparation of 7- [2- (2-aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamide] cephalosporin derivatives of the formula

<img file="CS264257B2_D0003.tif" />

where

R 2 NH is an amino group optionally protected by a

R 5 represents a hydrogen atom or a "nucleophilic compound residue".

The term "nucleophilic compound moiety", which has a broad meaning, refers to a variety of moieties, and it is also stated that R 3 may be a quaternary ammonium group. However, only the pyridinium moiety, the substituted, quinodonium, picolinium and lutidinium moieties as possible additions of quaternary amomi groups is exemplified. No mention is made of a quaternary ammonium group of the type comprising a compound of formula I. U.S. Pat. 1 U.S. Pat. No. 581,854 discloses a similar invention, in other patents, for example U.S. Patent No. 4,098,888 and its U.S. Patent Nos. 4,203,899, 4,205,180 and 4,298,606, and in British Patent No. 1,536,281 also similar compounds.

British Patent Application No. 2,040,921 discloses cephalosporin derivatives of the general formula

<img file="CS264257B2_D0004.tif" />

<img file="CS264257B2_D0005.tif" />

where

Rⁿ and R^b are (C 1 -C 4) alkyl radicals or together with the carbon atom to which they are attached a C 3 -C 7 cycloalkylidene ring; and

R¹, R² and R³ denote alkyl radicals having 1 to 4 carbon atoms.

The present invention relates to a process for the preparation of cephaloporin derivatives of the general formula (I) according to the invention.

<img file="CS264257B2_D0006.tif" />

(I) where

R¹ represents a hydrogen atom

R² and R³ are independently methyl or ethyl; and

R⁴ means methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, allyl, 2-butenyl, 3-butenyl-2-hydroxyethyl, 3-hydroxypropyl, 2- (dimethylamino) ethyl, pyridyl, methyl, pyridylethyl, benzyl or phenethyl as well as non-toxic, pharmaceutically acceptable acid addition salts of these compounds. Also included within the scope of the invention are the salts of the compounds of formula I, including hydrates, as well as the tautomeric forms of the compounds of formula I, for example the 2-iminothiazolin-4-yl form of the 2-aminothiazol-4-yl group.

As can be seen from the structural formula, the compounds of formula I have a "syn" or "Z" configuration relative to the alkoxyimino group. Because they are geometric isomers, a small amount of the anti-isomer may be present. The invention relates to compounds of the formula I containing at least 90% of the syn isomer. The compounds of formula I are preferably syn isomers substantially free of the corresponding anti-isomers.

Non-toxic, pharmaceutically acceptable acid addition salts are, for example, acid salts:

hydrochloric, hydrobromic, formic, nitric, sulfuric, methanesulfonic, phosphoric, acetic and trifluoroacetic acids, other acids can also be used, as commonly used with penicillin and cephalosporin derivatives.

Compounds of formula (I) wherein R 1¹means a hydrogen atom having high antibacterial activity against various Gram-positive and Gram-negative bacteria and can be used to treat bacterial infections in animals including humans. The compounds of formula (I) may be formulated for parenteral administration in a conventional manner using conventional pharmaceutical carriers and excipients, usually in the form of a unit dose, in multiple-dose kits.

The compositions may be solutions, suspensions, or emulsions in oily or aqueous media, and may contain conventional dispersing, stabilizing and suspending agents.

The compositions may also take the form of a dry powder to be reconstituted just prior to use, for example in sterile, pyrogen-free water.

The compounds of formula (I) may also be formulated into a suppository using a conventional base such as cocoa butter or other glycerides. The compounds of the present invention may also be administered in combination with other antibiotics, for example, other penicillin or cephalosporin derivatives, if desired.

When formulated into pharmaceutical compositions, a single dose usually contains between 50 and 1500 mg of an active compound of the formula I. The dosage of the compounds of the formula I depends on various factors, for example the weight and age of the patient and also the nature and severity of the disease. identify each doctor. A typical adult dose will usually be in the range of 500-5000 mg per day, depending on the frequency and route of administration. When a compound of formula I is administered intramuscularly or intravenously to an adult human, the total daily dose will be 750 to 3000 mg, usually given in divided doses, but higher doses of some of the compounds may also be administered, especially in cases of Pseudomonas infection.

[Preferred compounds of formula I are those wherein:

R¹ means a hydrogen atom,

R² means methyl,

R³ means methyl or ethyl,

R⁴ represents methyl, ethyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl, allyl or pyridylmethyl.

Particularly preferred compounds are those in which:

R¹ means a hydrogen atom,

R² and R³ are methyl radicals and

R⁴ represents methyl, 2-hydroxyethyl or allyl.

The most preferred compound of the above group is a compound in which

R¹ represents a hydrogen atom and

R², R³ and R⁴ are methyl radicals.

In a basic evaluation of the preferred compounds of the invention, the minimum inhibitory concentrations (MICs) for each compound were determined by conventional dilution series on agar, in which all experiments were performed twice, using Mueller-Hinton agar and 32 strains of the microorganisms studied in six groups. The geometric averages of the MIC values ​​determined in this manner are given in Table 1 below.

Table 1

Compound Geometric mean MIC µg / ml of Example No. (G +) - Ia [GjJ-Ib (G-) - Ia (G-) -Ib (G-) -II (G-J-III (5 strains) (5 (5) (6) (5) (6)

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The in vitro potency of the most preferred compound 1a against 31 strains of selected bacteria was determined on GC agar, the results are shown in Table 3 below.

Table 3

In vitro potency of compound Ia

Explanatory notes to Table 1;

(G ~ H-Ia: Penicillin-sensitive S. aureus (5 strains) (G +) Ib: Penicillin-resistant S. aureus (5 strains) (G-) -Ia: Cephalothin-sensitive E. coli (2 strains) , P. pneumoniae (1 strain) • and P. mirabilis (2 strains) (G -) - Ib: cephalothin resistant E. coli (3 strains) and P. pneumoniae (3 strains) (G-) -II: Pr morganii (1 strain), Ent. cloacae (2 strains) and Ser marcescens (2 strains) (G -) - III: Ps. aeruginosa (6 strains) ^(at) Two different samples of compound Ia.

The absorption of the most preferred compounds of formula Ia, especially the compounds prepared according to Example 1, was determined in mice after a single intramuscular injection of the test substance, which was dissolved in 0.1 M phosphate buffer at pH 7, at a dose of 20 mg / kg. Blood samples were collected from orbital venous plexus into heparinized capillaries, and tests were performed in Mueller-Hinton environment using Morganella morganii A9696 as the test organism. The following table 2 shows the blood concentrations after different time, half - life (ti_/2) and the area under the AUC curve.

Table 2

Blood level of Compound Ia

Minutes after administration Blood level in µg / ml

<td> 10</td><td> 14,0</td>

<td> 20</td><td> 12,0</td>

<td> 30</td><td> 8,8</td>

<td> 40</td><td> 7,5</td>

<td> 50</td><td> 4,7</td>

<td> 60</td><td> 4,4</td>

<td> 90</td><td> 1,5</td>

<td> 120</td><td rowspan="2"> 0,74</td>

<td>'Г—-</td>

<td>ti / n (minutes)</td><td> 24,0</td>

<td>AUC, ug. h / ml</td><td> 10,0</td>

Microorganism

Geometric mean MI (Ug / ml

<td>S. pyogenes (6 strains)</td><td> 0,013</td>

<td>S. pneumoniae (7)</td><td> 0,013</td>

<td>N. meningitidis (6)</td><td> 0,016</td>

<td>N. gonorrhoeae (3)</td><td> 0,013</td>

<td>H. influenzae (6)</td><td> 0,013</td>

<td>ampicillin sensitive</td><td></td>

<td>H. influenzae (2)</td><td> 0,20</td>

<td>ampica-resistant</td><td></td>

linu

The present invention relates to a process for the preparation of compounds of formula (I). There are two basic methods by which an easily obtainable starting cephalosporin can be converted to another cephalosporin having different substituents at the 7 and 3-positions. 7 and replaced with the desired substituent, and then introducing the desired substituent into the form 3. However, it is also possible to first introduce a substituent at the 3-position and then replace the substituent at the 7-position. The compounds of formula I can be prepared by both methods, both of which are within the scope of the invention a substituent at the 7-position and then introduce the substituent at the 3-position.

A more preferred procedure is shown in Reaction Scheme 1, the other of which is shown in Reaction Scheme 2. The abbreviation Tr stands for trityl (triphenylmethyl), which is a preferred amino-protecting group. Ph stands for phenyl. The group —CH (Ph) 2 is thus a benzhydryl group, which is a preferred protecting group on the carboxyl group.

Reaction Scheme 1

<img file="CS264257B2_D0007.tif" />

STEED²·

C-C00C, H₅ (II)

С-СООСЯí 5 '1

<img file="CS264257B2_D0008.tif" />

TrHN '»

N - - C - CONH

AND_with7a _n^WITH \ o0

OR ^

<img file="CS264257B2_D0009.tif" />

COOCH (Ph

Ί

1. CHfN &lt;

^{J 4}R · '

<img file="CS264257B2_D0010.tif" />

-C-CONH

II

N&gt;

STEED²*

·. Protective groups in

<img file="CS264257B2_D0011.tif" />

(AND)

4,

Reaction Scheme 2

<img file="CS264257B2_D0012.tif" />

(Vlil)

COOCH_of (IA)

Nile

<img file="CS264257B2_D0013.tif" />

PhCH2 CONH

<img file="CS264257B2_D0014.tif" />

COOCH (Ph), r 3 ch, -n '

R * (X)

AND^Q R * ^х Ф |

CH-N-CH.

£ I ³

Rb

COOCH_of

<img file="CS264257B2_D0015.tif" />

τθ *

I? 5 ®l (Xll) ^CHfN-CH_dRb

COOCH_of

IV v

C-COONM

II

N

STEED²-

<img file="CS264257B2_D0016.tif" />

~ r

AND

<img file="CS264257B2_D0017.tif" />

<img file="CS264257B2_D0018.tif" />

AND ^x ® ^!· CM - N - C f / o

I. AND

COOCH_and (λ III) f I has removed the ochchann ych sk &lt;tpč n

Θ &lt;,

IR³ ® i

CH-N-Cl-L * I ³

Rb

COOCH (Ph) ^ (All)

While preferred multi-step processes for the preparation of compounds of Formula I are given in the above reaction schemes, it is understood that other starting materials and processes may be used to produce intermediates used in the key step of each reaction scheme. A key step in Reaction Scheme 1 is the reaction of compound VII with a tertiary amine. Compound VII can be prepared by another method. A key step in Reaction Scheme 2 is the acylation of a compound of Formula VII with a compound of Formula XIII. Compounds of formulas XII and IV may be obtained by another method.

The invention relates to a process for the preparation of compounds of the formula I

<img file="CS264257B2_D0019.tif" />

<img file="CS264257B2_D0020.tif" />

No.

C- C-

<img file="CS264257B2_D0021.tif" />

R³And CH-NR3

<img file="CS264257B2_D0022.tif" />

where iR² and R³ are independently methyl or ethyl; and

R⁴ means:

methyl, ethyl, n-propyl, isopropyl, n-butyl, isoibutyl, allyl,

2-Butenyl-3-butenyl

2-hydroxyethyl,

3-hydroxypropyl,

2- (dimethylamino) ethyl, pyridylmethyl, pyridylethyl, benzyl or phenethyl, as well as non-toxic pharmaceutically acceptable salts and solvates thereof, characterized in that the compound of formula XIV is reacted

<img file="CS264257B2_D0023.tif" />

<img file="CS264257B2_D0024.tif" />

<img file="CS264257B2_D0025.tif" />

where _

R² has the above meaning,

(B)¹ represents a conventional carboxyl protecting group and

(B)² represents a conventional amine protecting group with a tertiary amine of formula

R³/ СНз — N where

R³ and R⁴ are as defined above to form a compound of formula XV

<img file="CS264257B2_D0026.tif" />

COOB (XV) *** «k._wi no

where

R², R³, R⁴, В¹ .a B² are as defined above with subsequent removal of all protecting groups in a conventional manner and / or optionally converting the compound of formula I so obtained into a non-toxic, pharmaceutically acceptable salt or solvate.

The reaction is carried out in a non-aqueous organic solvent such as methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile and the like, or mixtures of these solvents. The reaction temperature is usually in the range of -10 ° C to + 50 ° C, but is usually carried out at room temperature. At least 1 mole of tertiary amine must be used per mole of compound of formula (XIV), it is usually preferred to use a 50% to 100% excess tertiary amine.

A carboxyl protecting group suitable for use as substituent B¹in the above reaction, one of the commonly known groups for this use may be, for example, an aralkyl group such as benzyl, p-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (benzhydryl), furthermore alkyl groups such as tert-butyl, haloalkyl groups such as 2,2,2-trichloroethyl or other protecting groups on the carboxyl group as described in the literature, for example British Patent Specification No. 1 399 086. It is preferred to use groups which are readily removed by acid treatment. Particularly preferred protecting groups on the carboxyl group are benzhydryl and tert-butyl.

Amino protecting groups which can be used as substituent B² are also known in the art, for example trityl and acyl groups, for example chloroacetyl. Preferred are protecting groups which are readily removed by acid treatment, for example trityl.

Tertiary amines of general formula

R³ /

СНз-N where

R³ and R⁴ as used hereinbefore in the preparation of the compounds of formula (I) are commercially available or readily prepared by known methods.

The invention will be illustrated by the following preparations and examples.

Preparation 1 f I - ~ C ~ C 00C

H 'и

- ''^_ор2 (III)

Ethyl (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetate (IIIa)

A mixture of 5.00 g (10.9 mmol) of (Z) -2-hydroxyimino-2- (2-tritylaminothiazol-4-yl) acetate (II), 2.04 ml (32.8 mmol) of methyl iodide and

4.54 g (32.8 mmol) of potassium carbonate in 100 ml of anhydrous dimethylsulfoxide (DMSO3) was stirred at room temperature overnight and then poured into 250 ml of water. yield 5.15 g of the title compound, m.p. 115 DEG C. with decomposition.

NMR: d 'CDCl 3 ppm:

1.32 (3 H, t),

3.98 (3 H, s),

4.30 (2H, q), 6.42 (1H, s),

7.2 (1H, m);

7.25 (15 H, s).

Compound IIIb was prepared as described above, but ethyl iodide was used instead of methyl iodide.

<td colspan="3">Table</td>

<td>Compound</td><td>R²</td><td>Yield (%)</td>

<td>lila</td><td>methyl</td><td> 100</td>

<td>Illb</td><td>ethyl</td><td> 67</td>

<td>Explanatory notes:</td><td></td><td></td>

Melting point

Melting point ° C (literature)^ίυ

115 120 decomposition decomposition up to 98% ester was hydrolyzed without isolation ^(J) Tetrahedron, 34, 2233 (1978)

284257

Preparation 2

TrHN

<img file="CS264257B2_D0027.tif" />

C-COOH

II

N '' OR² (IV)

(Z) -2-Methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa)

6.00 g (12.7 mmol) of ethyl ester of formula IIIa prepared according to Preparation 1 in 120 ml of ethanol are treated with 12.7 ml of 2 N sodium hydroxide and left to stand overnight at room temperature. The residue was dissolved in 100 ml of water and the solution was acidified with 1N hydrochloric acid to pH 2 and then extracted with 3 X 50 ml of ethyl acetate. The extracts were combined, washed with a saturated aqueous sodium chloride solution, dried and then evaporated. The residue was crystallized from ethyl acetate / hexane to give the title compound (5.56 g, 98%), mp 138-143 ° C dec.

NMR: With CDCl 3 ppm:

3.89 (3 H, s),

6.52 (1 H, s),

7.2 (15H, s).

The compound of formula IVib was prepared in the same manner.

Merge- 'R² Nina yield (%)

Melting point ° C dec

Melting point ° C with decomposition from literature^{* 11}’

IVa methyl 98

IVb ethyl 85

Explanatory notes:

Tetrahedron, 34, 2233 (1978).

Preparation 3

Benzhydryl-3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate (VIII)

5 g (12.1 mmol) of 7-phenylacetamidocephalosporonic acid sodium salt are added in one portion to a stirred suspension of pH 7 phosphate buffer of 162.5 ml and 20 g of dried wheat bran at room temperature. The progress of the reaction was monitored by high pressure liquid chromatography until hydrolysis was complete (5 hours). Wheat bran is filtered from the suspension and the filtrate is cooled to 5-10 ° C for extractive esterification. To the cooled solution was added 32 mL of methylene chloride and then 24 mL of a 0.5 M diphenyldiazomethane solution in methylene chloride. The pH was then adjusted to 3.0 by adding 28% phosphoric acid. After 1 hour the temperature of the mixture was allowed to rise to 20 ° C. 56 ml of heptane are slowly added and the resulting crystalline product is collected by filtration. 3.0 g of product are obtained in a yield of 50%.

Preparation 4

Benzhydryl-7-amino-3-chloromethyl-3-cephem-4-carboxylate (V)

To a suspension of phosphorus pentachloride (8.3 g, 40 mmol) in methylene chloride (100 mL) was added pyridine (3.2 g, 40 mmol) and the mixture was stirred at 20 ° C for 20 min. Then join

138 to 143 140

140 To the mixture was added 5.1 g (10 mmol) of benzhydryl-3-hydroxymethyl-7-phenylacetamido-3-cephene-4-carboxylate, prepared according to Preparation No. 3, with stirring at -40 degrees Celsius. The mixture was stirred at -10 ° C for a further 15 minutes and then allowed to stand at -10 to -15^andC still 7 hours. To the solution cooled to -20 ° C was added 10 ml of propane 1,3-diol and the mixture was allowed to stand for 16 hours at -20 ° C and then for 20 minutes at room temperature with stirring.

The resulting solution was washed twice with 20 ml of ice-water and 10 ml of saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated in vacuo. 12 g of the thus obtained gummy residue are dissolved in a 2: 1 mixture of chloroform and n-hexane and subjected to chromatography using a 200 g silica gel column and the same solvent as the eluent. The fractions containing the resulting product were evaporated in vacuo and the residue triturated with n-hexane to give 2.1 g (51%) of product, mp &amp; gt; 110 ° C.

Infrared spectrum in potassium bromide pellets has maxima at 3 400, 2 800, 1 785, 1 725 cm¹.

The spectrum in ultraviolet light in ethanol has maxima at 265 nm (Ei _cm^1% 160).

NMR: With DMSO-d 6 + CDCl 3 ppm:

3.69 (2 H, s),

4.43 (2 H, s),

5.09 (1H, d, J = 4.5Hz),

5.24 (1H, d, J = 4.5Hz),

6.87 (1Ή, s),

7.3 (10 H, m).

Preparation 5

Benzhydryl 3-chloromethyl-7 - [(Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (VIa)

2.29 g (5.52 mmol) of benzhydryl-7-amino-3-chloromethyl-3-cephem-4-carboxylate, prepared as described in Preparation 4 in 57 ml of methyl cyanide, are mixed with 4.09 ml (16.6 mmol). bis (trimethylsilyl) acetamide (BSA) at room temperature for 50 minutes, after which time a clear solution was formed. To this solution is added an acid chloride solution prepared from 2.04 g (4.60 mmol) of (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa) and 1 M 15 g (5.52 mmol) of phosphorus pentachloride in 20 ml of methylene chloride. The mixture was stirred at room temperature for 30 minutes, then poured into 200 ml of cold water and extracted three times with 100 ml of ethyl acetate. The extracts were combined, washed with brine, dried and evaporated.

The g of syrup residue is chromatographed on a column containing 150 g of silica gel, eluting successively with a mixture of toluene and ethyl acetate first in a ratio of 10: 1 and then in a ratio of 3: 1. The fractions containing the desired product were combined and evaporated to give 2.61 g of the compound of formula VIa as an amorphous powder in a yield of 68%.

NMR: δ CDCl3 ppm:

3.50 (2 H, s),

4.02 (3 H, s),

4.33 (2 H, s),

4.98 (1 H, d),

5.87 (1 H, q),

6.65 (1 H, s),

6.90 (1 H, s),

7.3 (25 H, m).

Preparation 6

Benzhydryl-3-iodomethyl-7 - [(Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (VIIa)

A mixture of 1.50 g (1.79 mmol) of the 3-chloromethylated derivative prepared according to Preparation 5 (VIa) and 1.34 g (8.93 mmol) of sodium iodide in 30 ml of methyl ethyl ketone was stirred at room temperature for 1 hour. After evaporation of the solvent, the residue was dissolved in 100 ml of ethyl acetate, washed with water, aqueous sodium thiosulphate solution and brine, dried and evaporated to give 1.47 g of the title compound (VIIa) as an amorphous powder in 89% yield.

NMR: δ CDCl3 ppm:

3.55 (2H, ABq);

4.00 (3 H, s),

4.25 (2 H, s),

4.97 (1 H, d),

5.80 (1 H, q),

6.65 (1 H, s),

6.90 (1 H, s),

7.3 (25 H, m).

Preparation 7

Benzhydryl-3-chloromethyl-7 - [(Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) Jacetamido] -3-cephem-4-carboxylate (VIb)

To a solution of 1.095 g (2.4 mmol) of (Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVb) in 20 mL of dichloromethane was added 500 mg of phosphorus pentachloride. The mixture was stirred at room temperature for 1 hour and then added to 1.083 g (2.4 mmol) of a solution of compound V and 1 ml of BSA in 20 ml of dichloromethane all at once, while cooling the solution with ice. The mixture was stirred for 2 hours and then poured into 200 mL of 10% aqueous sodium bicarbonate and extracted with 100 mL of chloroform. The extract was washed with water, dried over magnesium sulphate and evaporated under reduced pressure. The residue is chromatographed on a silica gel column. Elution with chloroform afforded 1.76 g of the product of formula VIb as an amorphous powder in a yield of 86%.

NMR: δ CDCl3 ppm:

1.40 (3H, t, CH 2 CH 3),

3.53 (2H, ABq, 2-NH),

4.37 (2H, s, -CH 2 Cl),

4.60 (2H, q, -N2СN3),

4.90 (1 H, d, 6-H),

5.89 (1 H, d, 7-H),

6.88 (1H, s, thiazole-H),

6.91 (1 H, s, benzhydryl-CH).

Preparation 8

Diphenylmethyl-7 - [(Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate (VIIb)

A mixture of 1.07 g (1.25 mmol) of VIb, prepared according to Preparation 7, and 562 mg (2.75 mmol) of sodium iodide in 20 ml of acetone were stirred for 1 hour. The mixture was filtered and the filtrate was poured into water and extracted with ethyl acetate. The organic layer was washed sequentially with 5% aqueous sodium thiosulphate solution, water and saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 1.04 g (89%) of VIIb.

NMR: δ CDCl 3 ppm:

3.55 (2H, q, 2H);

4.27 (2H, s, CH 2), 5.02 (1H, d, 6-H),

5.87 (1H, d, 7-H),

6.68 (1H, s, thiazole-H),

6.93 (1H, s, benzhydryl-CH).

Example 1

7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (trimethylammonium) methyl-3-cephem-4-carboxylate (Ia) ml (1 mmol) of a solution of trimethylamine in diethyl ether at a concentration of 1M, benzhydryl-3-iodomethyl-7 - [(Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem- Of 4-carboxylate (Fairy) in 30 ml of diethyl ether and stirred for 1.5 hours. 410 mg of precipitated quaternary salt (XVa) was collected by filtration in a yield of 82% and 3 ml of trifluoroacetic acid (TFA) were added. Mixture^:The mixture is stirred at room temperature for 1.5 hours and then evaporated to dryness under reduced pressure at less than 20 ° C. The residue was triturated with ether and the precipitated trifluoroacetic acid salt was collected by filtration in a yield of 365 mg, dissolved in a small amount of methanol and chromatographed on a 1.8 X 20 cm column packed with IHP-20 resin. The column was washed with 1 L of water and then eluted with 0.5 L of 30% aqueous methanol.

The methanol eluate was evaporated under reduced pressure at less than 40 ° C and the residue was lyophilized to give 129 mg of the crude product. Ratio Λ²/ Δ³ of the isomers in the crude product is 1: 2 as shown by high pressure liquid chromatography. In the same way, the product was also purified using Lichrosorb RP-18 on an 8 X X 300 mm column eluting with 1/100 M ammonium dihydrogen phosphate (pH 7.2: methanol = 85:15). The eluate is chromatographed on a 1.8 X 15 cm HP-20 resin column to remove the inorganic salt. The column was washed with 0.5 L of water and then with 0.5 L of 30% aqueous methanol.

The methanol eluate was evaporated under reduced pressure at less than 40% and the residue was lyophilized to give the title compound Ia as an amorphous powder. Yield = 33%, 75 mg based on compound VIIa. The product decomposes gradually at a temperature higher than 160 ° C. Purity is 80%.

The infrared spectrum in potassium bromide pellets has peaks at 3600 to 3000, 1775, 1660, 1610, 1440, 1350, 1030.

The ultraviolet absorption spectrum in 1/15 M phosphate buffer at pH 7 has maxima at 235 (ε 15,700), 257 (ε 15,400).

NMR: δ D2O ppm:

3.25 [9H, s, N + (S)],

4.10 (3H, s, OD),

5.47 (1H, d, 4Hz, 6-H),

5.96 (1H, d, 4Hz, 7-H),

7.10 (1H, s, thiazole-H).

Example 2

7- [(Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- [N, N-dimethyl-N- (2-hydroxyethyl-Jamonium) methyl-3-cephem-4- carboxylate (Ib)

The process of Example 1 was repeated except that an equimolar amount of N, N-dimethylethanolamine was used instead of trimethylamine. The crude product has a ratio of Δ²/ Δ³After purification, the resulting product was obtained in 17% yield and decomposed above 160 ° C. The purity determined was 90%.

Example 3

7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (N-dimethyl-N-allylammonium) methyl 3-cephem-4-carboxylate (Ic)

The process of Example 1 was repeated except that an equimolar amount of N, N-dimethylallylamine was used instead of trimethylamine. The crude product has a ratio of Δ²:: Δ³ isomer 1: 4.5. After purification, the resulting product was obtained in 14% yield and decomposed above 150 ° C. The purity determined is 80%.

Example 4

7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- [N, N -dimethyl-N- (3-pyridylmethyl) ammonium] methyl-3-cephem- 4-carboxylate (Id)

The process of Example 1 was repeated except that an equimolar amount of 3- (dimethylaminomethyl) pyridine was used instead of trimethylamine. The crude product has a ěr²: Δ³ isomer 1: 4.3. After purification, the resulting product was obtained in a yield of 17% and decomposed above 170 ° C. The purity determined was 75%.

Example 5

7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- [N, N-dimethyl-N- (2-dimethylaminoethyl jamonium) methyl-3-cephem-4- carboxylate (Ie)

The process of Example 1 was repeated except that an equimolar amount of 1,2-bis (dimethylamino) ethane was used in place of trimethylamine.² : Δ³ isomer 1: 1. After purification, the resulting product was obtained in 14% yield and decomposed above 150 ° C. The purity determined is 65%.

Example 6

7- [(Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (N, N-dimethyl-N-ethylammonium) methyl 3-cephem-4-carboxylate (If)

The process of Example 1 was repeated except that an equimolar amount of N, N-dimethylethylamine was used instead of trimethylamine. The crude product has a ratio of Δ²:: Δ³ isomer 1: 1. After purification, the resulting product was obtained in 15% yield, and decomposed at a temperature greater than 150 ° C. The purity determined was 77%.

Example 7

7- [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (N, N-diethyl-N-methylammonium] methyl-3-cephem-4-carboxylate (Ig j

The process of Example 1 was repeated except that an equimolar amount of diethylmethylamine was used instead of trimethylamine. The crude product has a ratio of Δ²: Δ³isomer 1: 1. After purification, the resulting product was obtained in 10% yield and decomposed above 150 ° C. The purity determined was 65%.

Example 8

7 - [(Z) -2- (2-aminothiazol-4-yl) -2-ethoxyiminoacetamido] -3- (trimethylammonium) methyl-3-cephem-4-cariboxylate (Ih)

The process of Example 1 was repeated except that benzhydryl-3-iodomethyl-7 - [(Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4- The carboxylate (VIIa) is replaced by an equimolar amount of the corresponding ethoxyimino compound (VIIb) prepared according to Example 8. The crude product has a ratio of Δ²: Δ³ 3: 1 isomer. After purification, the resulting product is obtained in a yield of 3% and decomposes above 150 ° C. The purity determined was 70%.

Spectral data for Examples 2 to 8

(a) Infrared spectrum in pellets with potassium bromide

All resulting products had a similar spectrum in infrared light:

770 up to 1,775 cm '¹ (, β-lactam C = O), 1660 cm-¹ (CONH),

610 cm '¹ (COO-).

b) Spectrum in ultraviolet light (1 / 15M phosphate buffer pH 7)

All products except Example 4 had similar spectra:

235 , nm (ε 15 700 to 16 400),

257 nm (ε 15 400 to 16 000).

The compound of Example 4 had the following values:

235 nm (ε 17 600),

255 nm (ε 18,600, sh)

260 nm (ε 19,000),

266 nm (ε 17,900, sh).

c) NMR spectra (D2O) for the compounds of each example are given in the following table:

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The ratio Δ²: Δ³ isomer and the yield of the resulting product can be improved by allowing the filtrate from the precipitated quaternary salt of formula XV to stand for a short period of time and then filtering again through a layer of compound XV, especially if this repeated filtration is repeated several times. The yield can also be increased somewhat by adding a small amount of the original tertiary amine to the filtrate. This is emphasized in the following example, which is a preferred embodiment of the method of Example 1.

Example 9

7- p (Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (trimethylammonium) methyl 3-cephem-4-carboxylate (1a)

A solution of trimethylamine in ether (10 mL of a 1 M solution) was added all at once with stirring to 4.68 g (5 mmol) of iodide compound VIIa in 500 mL of ether. The mixture was stirred for 10 minutes and the precipitated quaternary salt of formula XVa was collected by filtration and washed with a small amount of ether. The filtrate and wash were combined and allowed to stand at room temperature for an additional 10 minutes, the second precipitate was collected by filtration using the same funnel, leaving the first precipitate as a layer. The filtrate is filtered three times in the same manner at 10 minute intervals to give 3.58 g of the quaternary salt of formula XVa in a yield of 72% relative to compound VIIa. To the resulting filtrate was added 2 ml of a 1 M solution of trimethylamine in ether, and the mixture was filtered again three times at 10 minute intervals, yielding 0.56 g of the compound of formula XVa in 11% yield. 1 ml of 1 M trimethylamine solution was added to the resulting filtrate, and the mixture was filtered again twice at 10 minute intervals to give 0.134 g of compound XVa as a third crop (2.7%). In total, 4.27 g of compound XVa is obtained in a yield of 86%.

A mixture of 4.20 g (4.22 mmol) of XVa, 1 ml of anisole and 40 ml of trifluoroacetic acid was stirred at room temperature for 1.5 hours. The mixture was evaporated under reduced pressure at less than 20 ° C and the dark residue was triturated with 300 ml of isopropyl ether to precipitate 3.50 g of the trifluoroacetic acid salt which was collected by filtration and dried under reduced pressure. The salt obtained is dissolved in 50 ml of methanol, a small amount of activated carbon is added and the mixture is filtered. The filtrate was concentrated under reduced pressure, and 15 ml of a 1 M solution of sodium 2-ethylhexanoate in ethyl acetate was added to the concentrate. The mixture was diluted with 300 ml of ethyl acetate to precipitate 2.36 g of the title product of 50% purity. This product was collected by filtration, washed with a small amount of ethyl acetate and dried. Ratio Δ² isomer Δ³ of the isomer in the crude product was 1: 4 as determined by high pressure liquid chromatography on LiChrosorb RP-18 mobile phase (a mixture of 1/100 M ammonium phosphate buffer pH 7 and methanol 90:10, retention time Δ² isomer 6 minutes 54 seconds, Δ³ isomer 8 minutes and 29 seconds.

2.36 g of the crude product was dissolved in a small amount of water and purified using high pressure liquid chromatography (Waters Associates, System 500, PrepPAK 500/18, mobile phase 7% methanol). The product-containing eluate was concentrated under reduced pressure at less than 35 ° C and the concentrate was lyophilized to give the title compound (959 mg, 42%), based on iodide compound VIIa. The product is an amorphous powder having a purity of 80% as determined by HPLC. Share Δ² isomer Δ³ The isomer is 1: 17. The product decomposes gradually at a temperature higher than 160 ° C.