Process for preparing new derivatives of cephalosporine

(54) Description New cephalosporin derivatives, process for their preparation and compositions thereof (56) Published publications: GB 2 040 921, US 4 278 671 (57) Abstract:

The invention relates to compounds of the formula

<img file="SE453092B_D0001.tif" />

<img file="SE453092B_D0002.tif" />

wherein R is hydrogen. R and R are each independently methyl or ethyl and R is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable acid addition salts and solvates thereof as well as processes for their preparation. These compounds are potent antibacterial agents.

D6 847289

453 092

The invention relates to novel cephalosporin derivatives of the formula

<img file="SE453092B_D0003.tif" />

<img file="SE453092B_D0004.tif" />

wherein r! is hydrogen, R² and R³ each is independently methyl or ethyl and R is⁴ are methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable acid addition salts or solvates thereof. The invention also relates to processes for the preparation thereof, and to antibacterial compositions.

British patent 1,399,086 generically discloses a large number of cephalosporins of the formula

<img file="SE453092B_D0005.tif" />

COOH wherein R is hydrogen or an organic group, R^A is an etheric monovalent organic group which is bonded to oxygen via a carbon atom, B is S or SO, and P is an organic group. However, the 2 aminothiazole-4-yl group is not identified as an R substituent and there are no examples where P is a quaternary ammonium methyl group of the type included in the compounds of formula I above. U.S. Patent Nos. 3,971,778, 4,024,133, 4,024,137, 4,064,346, 4,033,950, 4,079,178, 4,091,209, 4,092,477, and 4,093,803 have similar content.

U.S. Patent 4,278,793 discloses

<img file="SE453092B_D0006.tif" />

453 092 generically a large number of cephalosporin derivatives of the formula

<img file="SE453092B_D0007.tif" />

COOR₃ wherein the substituents R ^, R.₂, R 2, R 2, X and A generically comprise corresponding substituents in the compounds of formula I above. However, in the 20-long definition of the various substituent groups, in the 78-page enumeration of structural formulas, and in the 225 embodiments, there is no disclosure that A could be a quaternary ammonium methyl group of the type included in the compounds of formula I above. British patent specification 1,604,971 corresponds to said US patent and has a substantially identical content. The published British patent application 2,028,305 A has the same broad generic content, although it is obviously not formally related to the aforementioned patents, but A is exemplified only as hydrogen.

U.S. Patent 4,278,671 discloses

7- (2- (2-Aminothiazol-4-yl) -2- (syn) -methoxyiminoacetamido) cephalosporin derivative of the formula

<img file="SE453092B_D0008.tif" />

where R₂NH is an optionally protected amino group and R₃ is hydrogen or the residue of a nucleophilic compound. The term residue of a nucleophilic compound encompasses a broad definition and it is thus stated that R₃ alternatively may be a quaternary ammonium group. Only pyridinium, on

453 092 different ways substituted pyridinium, quinolinium, picolinium and lutidinium are revealed as possible quaternary ammonium groups. There is some suggestion that the quaternary ammonium group may be of the type included in the compounds of formula I above. British Patent Specification 1,581,854 corresponds to said US Patent Specification and has substantially identical content. Other patents in the name of the same patent holder, which are not formally related to the aforementioned patent, but which have similar contents, include U.S. Patents 4,098,888, 4,203,899, 4,205,180 and 4,298,606 and British Patent 1,536,281.

Published British Patent Specification 2,040,921 discloses cephalosporin derivatives of the formula

<img file="SE453092B_D0009.tif" />

the carbon atom to which they are attached forms a C1-6 cyclo123 alkylidene ring and R, R and R each are a C1-6 alkyl group.

The present invention relates to cephalosporin derivatives of the formula

R¹ HN ^ S '^ OR² wherein is hydrogen, R² and R²is methyl or ethyl and R⁴

<img file="SE453092B_D0010.tif" />

each is independently methyl, ethyl, allyl,

453 092

2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable acid addition salts thereof, as well as processes for their preparation. Also included within the scope of the present invention are solvates (including hydrates) of the compounds of formula I 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 shown in the structural formula above, the compounds of formula I have a syn or Z configuration with respect to the alkoxyimino group. Since the compounds are geometric isomers, some amount of the anti-isomer may also be present. The invention comprises compounds of formula I containing at least 90% of the syn isomer. Preferably, the compounds of formula I are syn isomers which are substantially free of the corresponding anti isomers.

The non-toxic, pharmaceutically acceptable acid addition salts of the compounds of formula I include the salts of hydrochloric, hydrobromic, formic, hydrochloric, sulfuric, methanesulfonic, phosphoric, acetic and trifluoroacetic and other acids used in the penic acid and penicillic acid.

in

The compounds of formula I wherein R is hydrogen exhibit high antibacterial activity with respect to various Gram-positive and Gram-negative bacteria and are useful for treating bacterial infections in animals including humans. The compounds of formula I may be formulated for parenteral use in the conventional manner using known pharmaceutical carriers and excipients and may be in unit dosage form or in multiple dosage containers. The compositions may be in the form of solutions, suspensions or emulsions in oily or aqueous carriers and may contain conventional dispersing, suspending or stabilizing agents. The compositions may also be in the form of a dry powder for reconstitution prior to use, for example with sterile pyrogen-free water. The compounds of formula I can also be prepared as supositories using conventional supository bases such as cocoa butter or other glycerides. The compounds of the present invention may, if desired, be administered in combination with other antibiotics such as penicillins or other cephalosporins.

When the compositions are in unitary form, they preferably contain 50-1500 mg of the active ingredient of formula I. The dosage of the compounds of formula I is dependent on such factors as the weight and age of the patient as well as on the particular nature and severity of the disease and is ultimately determined by physician. However, the dosage for treatment of adult humans is in the range of 50-5000 mg per day, depending on the frequency of administration and the mode of administration. In intramuscular or intravenous administration to adults, a total dose of 750-3000 mg per day, in divided doses, is sufficient, although higher daily doses of some compounds may be desirable for Pseudomonas infections.

The preferred compounds of formula I are those wherein 12 is 4

R is hydrogen, R is methyl, R is methyl or ethyl and R is methyl, ethyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl, allyl or pyridylmethyl. Particularly preferred compounds are those wherein R is hydrogen, R and R are methyl and R is methyl, 2-hydroxyethyl or allyl. The most preferred compound is R 2 is hydrogen and R, R, R are methyl. In the preliminary evaluation of the preferred compounds of the present invention, the minimum inhibitory concentrations, (MIK) of the compounds were determined by the two-fold agar series dilution method of Mueller-Hinton agar with respect to 32 strains of the test organisms in 6 groups. The geometric mean values ​​of MIK data for this test are presented in Table 1 below.

453 092

Chart

<td></td><td>HH H ~ ¹ s T 0</td><td>O ear OH X cn</td><td>in • vn</td><td> 5,6</td><td>X &gt;</td><td colspan="2">os X r-</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>os</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td> \0</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>hrs</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>H -x</td><td>O</td><td>O</td><td>cn</td><td>os</td><td></td><td></td>

<td></td><td>im</td><td></td><td>tO</td><td>to</td><td>to</td><td></td><td></td>

<td></td><td>X *</td><td>os</td><td>X</td><td>X</td><td>X</td><td>X</td><td></td>

<td></td><td> 1</td><td>t ©</td><td>O</td><td>O</td><td>O</td><td>V "</td><td></td>

<td></td><td>ώ</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>g</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>ζρ</td><td></td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td>a.</td><td></td><td>CN</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>X)</td><td> *</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>HRS --</td><td>O</td><td>CO</td><td> 04</td><td> 04</td><td>O</td><td></td>

<td rowspan="2"></td><td>1 tD</td><td> «*</td><td>r</td><td> 04</td><td> 04</td><td>in</td><td></td>

<td>z¹*· ***</td><td>m</td><td></td><td>X</td><td>X</td><td>X</td><td></td>

<td>g</td><td> 1</td><td>Oh</td><td>O</td><td>O</td><td>O</td><td>O</td><td></td>

<td></td><td> 0</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td rowspan="2">e for</td><td></td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td rowspan="2">Έ</td><td></td><td>os</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td> 04</td><td></td><td></td><td></td><td></td><td></td>

<td>ΣΓΠ</td><td></td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td> &gt;</td><td></td><td>X</td><td>CO</td><td>sr</td><td>r</td><td>r&gt;</td><td></td>

<td>RQ</td><td>t</td><td>O</td><td>n</td><td></td><td>tn</td><td> 00</td><td></td>

<td rowspan="2">Φ</td><td>HRS -</td><td>ear</td><td>Q</td><td>O</td><td>O</td><td>O</td><td></td>

<td>i tn</td><td>m</td><td>X</td><td>X</td><td>X</td><td>X</td><td></td>

<td>m</td><td></td><td>m</td><td>O</td><td>O</td><td>O</td><td>O</td><td></td>

<td>g</td><td> 1</td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>island</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td>4J</td><td></td><td>O</td><td></td><td></td><td></td><td></td><td></td>

<td>λ:</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>tn</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>LI</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>-u</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td> 0</td><td>β-</td><td>u&gt;</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>i tn</td><td>m</td><td>Γ</td><td>to</td><td>· Γ</td><td>m</td><td></td>

<td></td><td>_--_hrs W</td><td>«X</td><td>X</td><td>X</td><td>X</td><td>X</td><td></td>

<td></td><td> +</td><td>to</td><td>sr</td><td>m</td><td>sr</td><td>m</td><td></td>

<td></td><td> 0</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>m</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>Έ</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>ti s</td><td>to</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>H ra</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>i 4-1</td><td></td><td>to</td><td>sr</td><td>T ~</td><td></td><td></td>

<td></td><td>~ ω</td><td>• X</td><td>X</td><td>X</td><td>X</td><td>X</td><td></td>

<td></td><td></td><td>V</td><td></td><td>V "</td><td> 04</td><td>OH</td><td></td>

<td></td><td>ϋ m</td><td>X</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>tPA</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>c</td><td>κ</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>Φ</td><td>t</td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>δ</td><td> •</td><td>v</td><td>OH</td><td>CQ</td><td>sr</td><td> 00</td><td></td>

<td>M</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>:O</td><td>c</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>t.</td><td>Φ</td><td></td><td></td><td></td><td></td><td></td><td></td>

<img file="SE453092B_D0011.tif" />

<td>β 1</td><td>β 1</td><td>Η Η 1</td>

<td></td><td> 1</td><td></td>

<td> 1 0</td><td>1 ο</td><td> 1 0</td>

453 092

The absorption of the most preferred compound Ia (prepared according to Example 1) was determined in mice following a single intramuscular injection of the test compound (dissolved in 0.1M phosphate buffer; pH 7) at a dose of 20 mg / kg. Blood samples were withdrawn from the eyelids and collected in heparinized capillaries and analyzed in MuellerHinton medium using Morganella morganii A9695 as the test organism. Blood levels at different time intervals, half-life values^oc^ Y^plates below the curve (YUK) is shown in Table 2 below.

Table 2

Blood levels of compound Ia in mice

<td>Blood levels pg / ml at minutes after administration</td><td> 10 20 30 40 50 60 90 120</td><td> 14 12 8,8 7,5 4,7 4.4 1.5 0,74</td>

<td>^t1/2 (minutes)</td><td></td><td> 24</td>

<td>YUK</td><td></td><td></td>

<td>pg-hr / ml</td><td> 1</td><td> 10</td>

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The in vitro activity of the most preferred compound Ia with respect to 31 strains of difficult to treat bacteria was determined in large GC agar and the results are presented in Table 3 below.

Table 3

In vitro activity of compound Ia with respect to difficult-to-treat bacteria

<td>test organism</td><td>Geometric mean for MIK (pg / ml)</td>

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

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

<td>N. gonorrhoeae (4)</td><td> 0,013</td>

<td>N. meninqitidis (5)</td><td> 0,016</td>

<td>H. influenzae (7) (Ampicillin-sensitive)</td><td> 0,013</td>

<td>H. influenzae (31 (Ampicillin resistant)</td><td> 0,20</td>

453 092

In another aspect, the invention relates to processes for the preparation of the compounds of formula I. There are two main methods for converting an easily accessible starting cephalosporin to another cephalosporin with various substituents at the 7- and 3-positions. One can first remove the 7-substituent and replace it with the desired 7-substituent and then insert the desired 3-substituent. Alternatively, one may first introduce the desired 3-substituent and then replace the 7-substituent. The compounds of formula I can be prepared by either process and both fall within the scope of the present invention, but it is preferred to introduce the desired 7-substituent first and then introduce the desired 3-substituent. The preferred process is presented in Scheme 1 below, while the alternative procedure is presented in Scheme 2 below. The abbreviation Tr represents the trityl group (triphenylmethyl), which is a preferred amino protecting group. The abbreviation Ph represents the phenyl group. Thus, -CH (Ph) is 2^_the benzhydryl group, which is a preferred carboxyl protecting group.

Scheme 1

<img file="SE453092B_D0012.tif" />

II

<img file="SE453092B_D0013.tif" />

<img file="SE453092B_D0014.tif" />

COOC₂hrs₅

III

OH

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TrHN

<img file="SE453092B_D0015.tif" />

COOH

IV

<img file="SE453092B_D0016.tif" />

<img file="SE453092B_D0017.tif" />

<img file="SE453092B_D0018.tif" />

After I

<img file="SE453092B_D0019.tif" />

<img file="SE453092B_D0020.tif" />

deprotection

VII

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<img file="SE453092B_D0021.tif" />

Scheme 2

<img file="SE453092B_D0022.tif" />

VIII

PC1₅pyridine

<img file="SE453092B_D0023.tif" />

IX

<img file="SE453092B_D0024.tif" />

Nal

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<img file="SE453092B_D0025.tif" />

<img file="SE453092B_D0026.tif" />

<img file="SE453092B_D0027.tif" />

PhC₂CON

<img file="SE453092B_D0028.tif" />

COOCH (Ph)₂^R deacylating

<img file="SE453092B_D0029.tif" />

XI

XII

IV

V

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XIII

<img file="SE453092B_D0030.tif" />

deprotection

<img file="SE453092B_D0031.tif" />

Although the above reaction schemes show preferred multistage processes for preparing the compounds of formula I, it will be appreciated that other starting materials and the process can be used to prepare the intermediates used in the key step of each reaction scheme. Thus, the key step in Scheme 1 is the reaction of compound VII with the tertiary amine. Compound VII may itself be prepared by other processes. Likewise, the key step in Scheme 2 is the acylation of Compound XII with Compound IV. Both compounds XII and IV can be prepared by other processes.

The present invention provides a process for the preparation of compounds of the formula

453 092

<img file="SE453092B_D0032.tif" />

<img file="SE453092B_D0033.tif" />

wherein R² and R² each is independently methyl or ethyl and R is⁴ are methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable salts and solvates thereof, which means reacting a compound with the

<img file="SE453092B_D0034.tif" />

<img file="SE453092B_D0035.tif" />

XIV wherein R is as defined above meaning 2 carboxyl protecting group B term

<img file="SE453092B_D0036.tif" />

<img file="SE453092B_D0037.tif" />

is a conventional is a conventional amino protecting group, with a tertiary amine of the formula

<img file="SE453092B_D0038.tif" />

4 wherein R and R have the same meanings, for the preparation of a compound of formula

<img file="SE453092B_D0039.tif" />

and then removing all protecting groups in a conventional manner.

The reaction is carried out in an anhydrous organic solvent such as methylene chloride, chloroform, ethyl ether, hexane.

453 092 ethyl acetate, tetrahydrofuran, acetonitil and the like or mixtures of such solvents. The reaction is preferably carried out at a temperature of from -10 ° C to + 50 ° C; normally it is preferred to carry out the reaction at room temperature. At least one mole of the tertiary amine should be used per mole of compound XIV; normally it is preferred to utilize 50-100% excess of the tertiary amine.

Carboxyl protecting groups suitable for use as B in the above reaction are well known to those skilled in the art and include aralkyl groups such as benzyl, p-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (benzhydryl); alkyl groups such as p-butyl; haloalkyl groups such as 2,2,2-trichloroethyl and other carboxyl protecting groups described in the literature, for example in British Patent No. 1,399,086. It is preferred to use carboxyl protecting groups which can be easily removed by treatment with acid. Particularly preferred carboxyl protecting groups are the benzhydryl and t-butyl groups.

Amino protecting groups suitable for use as B are also well known in the art and include the trityl group and acyl groups such as chloroacetyl. Amino protecting groups which can be easily removed by treatment with acid, for example the trityl group, are preferred.

The present invention also includes a process for the preparation of compounds of the formula

O

<img file="SE453092B_D0040.tif" />

<img file="SE453092B_D0041.tif" />

3 wherein R and R are each independently methyl or ethyl and R 2 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethyl 453 092 amino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable salts and solvates thereof, which process means that one acylates a compound of the formula

<img file="SE453092B_D0042.tif" />

XVI or an N-silyl derivative thereof, wherein B is hydrogen or

4 a conventional carboxyl protecting group and R and R have the above meanings, with an acylating derivative of an acid of the formula

<img file="SE453092B_D0043.tif" />

"COOH

XVII

2 wherein B is a conventional amino protecting group and R is as defined above for the preparation of a compound of the formula

<img file="SE453092B_D0044.tif" />

and that all protective groups are then removed.

The acylating derivatives of the acid of formula XVII include the acid halides (and especially the acid chloride), mixed acid anhydrides (such as the acid anhydrides formed with pivalic acid or a haloformate such as ethyl chloroformate) and activated esters (such as may be formed with Nhydroxybenzstriazole in the presence of a condensing hydroxide dichloromethoxy). The acylation can also be carried out using the free acid of formula XVII in the presence of a condensing agent such as dicyclohexylcarbodiimide, carbonyldiimidazole or an isoxazolium salt. The term acylating derivative of the acid of formula XVII in the present context includes the free acid itself in the presence of a condensing agent as described above. The preferred acylating derivative of the acid of formula XVII is the acid chloride which is preferably used in the presence of¹ of an acid binding agent (and especially a tertiary amine as an acid binding agent such as triethylamine, dimethylaniline or pyridine).

When the acylation is carried out with an acid halide, it is possible to use an aqueous reaction medium but an anhydrous medium is preferred. When acid anhydrides, activated esters or the free acid in the presence of a condensing agent are used for the acylation, the reaction medium should be anhydrous. Particularly preferred solvents for the acylation reaction are halogenated hydrocarbons such as methylene chloride and chloroform, but tertiary amides such as dimethylacetamide or dimethylformamide as well as other conventional solvents such as tetrahydrofuran, acetonitrile and the like can also be used.

The acylation reaction can be carried out at a temperature of from -50 ° C to + 50 ° C. However, it is preferably performed at or below room temperature and especially at a temperature of -30 ° C to 0 ° C. It is usually preferred to acylate the compound of formula XVI with an approximately stoichiometric amount of the acylating agent of formula XVII, although a small excess (e.g., 5-25%) of the acylating agent may be used.

It is preferred that the compound of formula XVI be acylated in the form of its N-silyl derivative (using an anhydrous reaction medium). This is conveniently done in situ by simply adding a suitable silylating agent (for example, N, O-bistrimethylsilylacetamide) to the solution of compound XVI before the addition of the acylating agent of formula XVII. It is preferred to use about 3 moles of silylating agent per mole of compound XVI, although this is not critical. The Chile compound is removed

<img file="SE453092B_D0045.tif" />

I are commercially available or can be readily prepared by methods known in the art.

The invention is further illustrated by the following embodiment.

Preparation of starting material

Example A

TrHN

<img file="SE453092B_D0046.tif" />

III

Ethyl (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetate (IIla) __________________________________________________________

A mixture of 5.00 g (10.9 mmol) of ethyl (Z) -2-hydroxyimino-2- (2-tritylaminothiazol-4-yl) acetate (II), 2.04 ml (32.8 mmol) CH 2 I and 4.54 g (32.8 mmol) K₂^CO3 * ¹Dry dimethyl sulfoxide (DMSO) was stirred overnight at room temperature and then poured into 250 ml of water. The precipitate formed was collected by filtration, washed with water and dried to give 5.15 g of the title compound (quantitative yield), mp 115 ° C (dec.).

NMR: δ^CDC13 ppm 1.32 (3H, t), 3.98 (3H, s) t 4.30 (2H, q),

6.42 (1H, s), 7.2 (1H, m), 7.25 (15H, s).

453 092

Compound IIIb was prepared by the above general procedure but replacing the methyl iodide with ethyl iodide.

Compound R³ Yield (%) Mp (° C) Literature value (¹

Mp. (° C)

<td>Badly</td><td>methyl</td><td> 100</td><td>115 ° (sand) approx. 120 ° (sand)</td>

<td>IIIb</td><td>ethyl</td><td> 67</td><td> 97-98° *</td>

<td>* The ester</td><td>hydrolyzed</td><td>without</td><td>insulation</td>

⁽¹) Tetrahedron 34, (1978) 2233

Example B

<img file="SE453092B_D0047.tif" />

---- C OOH ^ OR²

IV (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa)

6.00 g (12.7 mmol) of the ethyl ester IIa prepared in Example A in 120 ml of ethanol were treated with 12.7 ml of 2N NaOH overnight at room temperature. The reaction mixture was adjusted to pH 8 by the addition of powdered dry ice and the solvent was evaporated under reduced pressure. The residue was dissolved in 100 ml of water and the solution was acidified with 1N HCl to pH 2 and then extracted with 3x50 ml of ethyl acetate. The combined extracts were washed with a saturated aqueous solution of sodium chloride, dried and evaporated. The residue was crystallized from ethyl acetate-hexane to give 5.56 g (yield 98%) of the title product, mp 138-143 ° C (dec.).

NMR:^CDC13 ppm 3.89 (3H, s), 6.52 (1H, s), 7.2 (15H, s).

Compound IVb was prepared by the above general procedure.

453 092

<td>Association</td><td>R²</td><td>Yield (%)</td><td>Mp (° C, sand)</td><td>literature value mp. (° C, sand)</td>

<td>IVa</td><td>methyl</td><td> 98</td><td> 138-143</td><td>ca. 140</td>

<td>IVb</td><td>ethyl</td><td> 85</td><td> 140-145</td><td>not reported</td>

⁽¹) Tetrahedron 34, (1978) 2233

Example C

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

To a stirred suspension of 162.5 ml of phosphate buffer (pH 7) and 20 g of dry wheat bran at room temperature was added 5 g (12.1 mmol) of 7-phenylacetamidocephalosporanoic acid sodium salt in one portion. The reaction was followed by HPLC until the hydrolysis was complete (5 hours). The suspension was filtered to remove wheat bran and the filtrate was cooled to 5-10 ° C for extractive esterification. To the cooled solution was added 32 ml of methylene chloride, followed by 24 ml of a 0.5 M solution of diphenyldiazomethane in ethylene chloride. The pH20 was then adjusted to 3.0 with 28% phosphoric acid.

After one hour, the reaction mixture temperature was allowed to rise to 20 ° C. 56 ml of heptane was added slowly and the resulting crystalline, title product recovered by filtration. The yield of the title product was 3.0 g (50%).

Example D

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

To a slurry of 8.3 g (40 mmol) of PC1₅ in 100 ml was added 3.2 g (40 mmol) of pyridine and the mixture was stirred for 20 minutes at 20 ° C. To the mixture was added 5.1 g (10 mmol) of the benzhydryl-3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate prepared in Example D in a portion with stirring at -40 ° C. The mixture was stirred at -10 ° C for 15 minutes and then allowed to stand at -10 ° C to -15 ° C for 7 hours. To the cooled solution (-20 ° C) was added 10 ml of propane-1,3-diol and the mixture was allowed to stand at -20 ° C for 16 hours and then 20 minutes at room temperature 453 092 with stirring. The resulting solution was washed with 2x20 ml of ice-water and 10 ml of a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated in vacuo. The gummy residue (12 g) was dissolved in a mixture of CHCl 3 and n-hexane (2; 1) and chromatographed using a silica gel column (200 g) and the same solvent as eluent. Fractions containing the title compound were evaporated in vacuo and the residue was triturated with n-hexane to give the title product (2.1 g; 51%) with a melting point greater than 110 ° C (dec.).

IR: 3400, 2800, 1785, 1725 cm¹.

UV EtOH _{265 (} 1% 150).

max. 1 cm

NMR: ^DMS0^d6 ^{+ CDC1}3 3.69 (2H, s), 4.43 (2H, s), 5.09 (1H, ppm d, J = 4.5Hz), 5.24 (1H, d, J = 4.5Hz),

6.87 (1H, s), 7.3 (10H, m).

Example E

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 the benzhydryl-7-amino-3-chloromethyl-3-cephem-4-carboxylate prepared in Example D in 57 ml of CH₃CN was treated with 4.09 ml (16.6 mmol) of bis (trimethylsilyl) acetamide (BSA) 50 minutes at room temperature to obtain a clear solution. To the solution was added an acid clotide solution prepared from 2.04 g (4.60 mmol) (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa) and 1.15 g (5). , 52 mmol) PC1,. in 20 ml of methylene chloride. The mixture was stirred for 30 minutes at room temperature, poured into 200 ml of cold water and extracted with 3x100 ml of ethyl acetate. The combined extracts were washed with aqueous sodium chloride solution, dried and evaporated. The residual syrup (4 g) was chromatographed on a silica gel column (150 g) by successively eluting with 10: 1 and 3: 1 mixtures of toluene and ethyl acetate. The fractions containing the desired compound were combined and evaporated to obtain

2.61 g (68%) of the compound Via as an amorphous powder.

<td>NMR: δ^CDC13 ppm 3.50</td><td>(2H,</td><td>s),</td><td> 4,02</td><td>(3H,</td><td>s),</td><td> 4,33</td><td>(2H,</td><td>s)</td>

<td> 4,98</td><td>(1H,</td><td>d)</td><td> 5,87</td><td>(1H,</td><td>q) '</td><td> 6,65</td><td>(1H,</td><td>s)</td>

<td> 6,90</td><td>(1H,</td><td>s),</td><td> 7,3</td><td>(25H,</td><td>m).</td><td></td><td></td><td></td>

Example F

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

A mixture of 1.50 g (1.79 mmol) of the 3-chloromethyl derivative (Via) prepared in Example E and 1.34 g (8.93 mmol) NaI in 30 ml of methyl ethyl ketone was stirred for one hour at room temperature. After evaporating the solvent, the residue was dissolved in 100 ml of ethyl acetate and washed with water, aqueous Na₂S₂O₂ and an aqueous solution of sodium chloride, dried and evaporated to give the title compound Vila (1.47 g, 89%) as an amorphous powder.

NMR

CDC1 ppm

3.55 (2H, ABg), 4.00 (3H, s), 4.25 (2H, s),

4.97 (1H, d), 5.80 (1H, q), 6.65 (1H, s), 6.90 (1H, s), 7.3 (25H, m).

Example G

Benzhydryl-3-chloromethyl-7-2 '(Z) -2-ethoxyimino-2- (2tritylaminothiazol-4-yl) acetamido7-3-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. After stirring for 45 hours at room temperature, the mixture was added in one portion to an ice-cooled solution of 1.083 g (2.4 mmol) of Compound V and 1 ml of BSA in 20 ml of dichloromethane. After stirring 0.5 hours, the reaction mixture was poured into 200 ml of a 10% aqueous sodium hydrogen carbonate solution and extracted with 100 ml of CHCl 3. The extract was washed with water, dried over magnesium sulfate and evaporated under reduced pressure. The residue was chromatographed on a silica gel column. Elution CHClg gave compound VIb as an amorphous

<td>powder in a variety</td><td>of 1,</td><td>, 76 g</td><td> (86%).</td><td></td>

<td>.CDC1, ___ NMR: δ 3 ppm</td><td> 1,40</td><td>(3H,</td><td>t, CH₂CH₃), 3.53 (2H,</td><td>ABq, 2-CH₂),</td>

<td></td><td> 4,37</td><td>(2H,</td><td>s, -CH₂C1), 4.60 (2H,</td><td>q, -CH₂CH₃),</td>

<td></td><td> 4,90</td><td>(1H,</td><td>d, 6-H), 5.89 (1H, d,</td><td>7-H), 6.88</td>

<td></td><td>(1H,</td><td colspan="2">s, thiazole-H), 6.91 (1H, s,</td><td>benzhydryl</td>

<td></td><td>CH).</td><td></td><td></td><td></td>

Example H

Diphenylmethyl-T - (C 2) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetamindo-5-iodomethyl-3-cephem-4-carboxylate (IIb) A mixture of 1.07 g ( 1.25 mmol) of the compound VIb prepared in Example G and 562 mg (2.75 mmol) NaI in 20 ml of acetone was 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 successively with a 5% aqueous Na solution₂S₂O₃, water and a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to give 1.04 g (89%) of compound VIIb.

<td> : &lt;5^CDC13 ppm 3.55</td><td>(2H,</td><td>q »</td><td>2-CH₂), 4.27 (2H, s, CH₂IN),</td>

<td> 5,02</td><td>(1H,</td><td>d</td><td>6-H), 5.87 (1H, d, 7-H),</td>

<td> 6,68</td><td>(1H,</td><td>s</td><td>thiazole ring H), 6.93 (1H,</td>

s, benzhydryl-CH).

453 092

Preparation of Compounds of the Invention

Example 1

7 - [(2) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamido] -3- (trimethylammonium) methyl 3-cephem-4-carboxylate (Ia) ml (1 mmol) of a 1M solution of trimethylamine in diethyl ether was added to a stirred suspension of 468 mg (0.5 mmol) benzhydryl-3-iodomethyl-7 - [(Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem -4-carboxylate (Vila) in 30 ml of diethyl ether and the mixture was stirred for 1.5 hours. The precipitated quaternary salt (XVa) was collected by filtration (410 mg; 82% yield) and 3 ml of trifluoroacetic acid (TFA) was added. This mixture was stirred for 1.5 hours at room temperature and then evaporated to dryness under reduced pressure at a temperature below 20 ° C. The residue was triturated with ether and the precipitated TFA salt was collected by filtration (yield 365 mg), dissolved in a small amount of methanol and chromatographed on a 1.8 x 20 cm HP-20 resin column. The column was eluted with about 1 liter of water and then with 0.5 ml of a 30% aqueous methanol solution. The methanolic eluate was evaporated under reduced pressure at a temperature below 40 ° C and the residue was freeze-dried to obtain the title crude product (yield 129 mg). The ratio of the Δ / Δ isomers in the crude product was 1: 2, as determined by HPLC. The product was purified by HPLC (Lichrosorb RP-18, 8x300 mm, elution with 1 / 100M NH₄hrs₂PO₄ (pH 7.2): CH₃OH = 85:15). The HPLC eluate was chromatographed on a column of HP-20 (1.8x15 cm) to remove the inorganic salt. The column was eluted with 0.5 L of water and then with 0.5 L of a 30% aqueous methanol solution. The methanolic eluate was evaporated under reduced pressure at a temperature below 40 ° C and the residue was freeze-dried to give the title compound (Ia) as an amorphous powder. Yield 75 mg (33%, based on Vila). The product was gradually decomposed above 160 ° C. Estimated purity 80%.

453 092

Example 2

7- (Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamido7-3 ~ ZN, N-dimethyl-N- (2-hydroxyethyl) ammonylmethyl-3cephem-4-carboxylate (Ib)

The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of Ν, Ν-dimethylethanolamine. The crude product had a Λ: Δ ratio of 1: 2. After purification, the title compound was obtained in a yield of 17% and decomposed at a temperature above 160 ° C. Estimated purity 90%.

Example 3

7 - [(Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamido-3- (N, N-diroethyl-N-allylamammonium) methyl 3-cephem-4-carboxylate (Ic)

The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of Ν, Ν-dimethylallylamine. The crude product had a Δ: / ratio of 1: 4.5. After purification, the title compound was obtained in 14% yield and decomposed at a temperature above 150 ° C. Estimated purity 80%.

Example 4

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

The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of 3- (dimethylaminomethyl) pyridine. The crude product had a Δ: A ″ ratio of 1: 4.3. After purification, the title compound was obtained in a yield of 17% and decomposed at a temperature above 170 ° C. Estimated purity 75%.

453 092 i

Example 5!

7-Z (Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamidoyl

-3 / 11, N-dimethyl-N- (2-dimethylaminoethyl) ammonium7metyl

-3-cephem-4-carboxylate (le);

The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced;

in an equimolar amount of 1,2-bis (dimethylamino) ethane. The crude product had a Δ: Δ ratio of 1: 1. After purification | The title compound was obtained in a yield of 14% and decomposed at a temperature above 150 ° C. Estimated purity 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 general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of Ν, Ν-dimethylethylamine. The crude product had a Δ: A ratio of 1: 1. After purification, the title compound was obtained in a yield of 15% and decomposed at a temperature above 150 ° C. Estimated purity 77%.

Example 7

1 - / (2) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamido /

-3- (N, N-diethyl-N-methylammonium) methyl 3-cephem-4-carboxylate (Ig)

The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of diethyl methylamine. The crude

3 had an A: Δ ratio of 1: 1. After purification, the title compound was obtained in 10% yield and decomposed at a temperature above 150 ° C. Estimated purity

65%.

453 092

Example 8

7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (ethoxyiminoacetamidoy-3- (triethylammonium) methyl 3-cephem-4-carboxylate (1h) The general procedure of Example 1 was repeated except) by substituting an equimolar amount of the corresponding ethoxyimino compound (VIIb) by substituting an equimolar amount of the corresponding ethoxyimino compound (V11 prepared according to Example H. The crude product had an A: Δ ratio of 3: 1. After purification, the title compound obtained 3% yield and decomposed at a temperature above 150 ° C. Estimated purity 70%.

Spectral data for the compounds of Examples 2-8 (a) Infrared Spectra (KBr)

All products exhibited similar infrared spectra: 1770-1775 cm¹ (β-lactam C = O), 1660 cm^-1 (CONH), 1610 cm¹(COO^-) .

(b) Ultraviolet spectra (1/15 M phosphate buffer; pH 7)

All products except the product of Example 4 exhibiting similar spectra: 235 nM (ε 15700-16400), 257 nM (_e 15400-16000). The compound of Example 4 exhibited the following values: 235 nM (ε 17600), 255 nM (ε 18600, sk), 260 nM (ε 19000), 266 nM (ε 17900, sk).

453 092 (c) NMR spectra (D₂O)

<td>Association</td><td colspan="4">Chemical displacement</td><td colspan="2">W</td>

<td>according to.</td><td></td><td></td><td>6-H</td><td>7-H</td><td></td><td></td>

<td>example</td><td>N⁺-Ql ·</td><td>O · OL</td><td>(1H, d,</td><td>(1H, d,</td><td>thiazole-H</td><td></td>

<td></td><td>(S) ³</td><td>(3H, S)</td><td>4-5Hz)</td><td>4-5ΗΖ)</td><td>(1H, s)</td><td>Other</td>

<td> 2</td><td>3.20 (3H) 3.25 (3H)</td><td> 4,10</td><td> 5,46</td><td> 5,90</td><td> 7,10</td><td></td>

<td> 3</td><td>3.08 (3H)</td><td> 4,10</td><td> 5,46</td><td></td><td> 7,10</td><td> 5,6-6,4</td>

<td></td><td>3.15 (3H)</td><td></td><td></td><td></td><td></td><td>(4H, m, 7-H, ch = ch₂)</td>

<td> 4</td><td>3.04 (3H)</td><td> 4,09</td><td> 5,45</td><td> 5,94</td><td> 7,06</td><td rowspan="2">4.75 (2H, s, PyrCH) 7.66 (TH, dd,</td>

<td></td><td>3.21 (3H)</td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td>8 &amp; 4 Hz, Pyr-H)</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td>8.12 (1H, d, 8 Hz, Pyr-H)</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td>8.74 (2H, s, ryr-H)</td>

<td> 5</td><td>3.18 (3H)</td><td> 4,10</td><td> 5,45</td><td> 5,90</td><td> 7,10</td><td>2.50 (6H, s,</td>

<td></td><td>3.25 (3H)</td><td></td><td></td><td></td><td></td><td>NfCH +)</td>

<td> 6</td><td>3.10 (3H)</td><td> 4,10</td><td> 5,45</td><td> 5,90</td><td> 7,10</td><td>1.48 (3H, t,</td>

<td></td><td>3.15 (3H)</td><td></td><td></td><td></td><td></td><td>7 Hz, CH₂CH₃)</td>

<td> 7</td><td>3.05 (3H)</td><td> 4,10</td><td> 5,44</td><td> 5,95</td><td> 7,10</td><td>1.45 (6H, t, 7 Hz, CH₂ol₃)</td>

<td> 8</td><td>3.25 (9H)</td><td></td><td> 5,50</td><td> 5,97</td><td> 7,10</td><td>1.45 (3H, t, 7 Hz, CH₂q₃)</td>

3

It has been found that both the Δ: Δ ratio and the yield of the product can be improved if the filtrate from the initially precipitated quaternary salt (XV) is allowed to stand for a short period of time and re-filtered through the bed of originally recovered compound (XV), especially if this re-filtration step is repeated. repeatedly. Thus, the yield can be slightly increased if a small amount of the original tertiary amine reactant is added to the filtrate. This is shown in the following example, which is a modification of Example 1.

453 092

Example 9

7 - [(2) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (trimethylammonium) methyl 3-cephem-4-carboxylate (Ia) 10 ml of a 1M solution of trimethyl amine was added one portion to a stirred solution of 4.68 g (5 mmol) of the iodide compound Vila in 500 ml of ether. The mixture was stirred for 10 minutes and the precipitated quaternary salt (XVa) was collected by filtration and washed with a small amount of ether. The filter funnel and the washing tubes were combined and allowed to stand at room temperature for a further 10 minutes and the second precipitate that was separated was collected by filtration on the same funnel where the first precipitate was kept as a bed. The filtrate was filtered three times in the same manner at 10 minute intervals to obtain. of 3.58 g (72% starting from Vila) of the quaternary salt (XVa). To the final filtrate was added 2 ml of a 1M solution of trimethylamine in ether and the same re-filtration operation was repeated 3 more times at 10 minute intervals to obtain 0.56 g (11%) of compound XVa. To the resulting filtrate was added an additional 1 ml of a 1M trimethylamine solution and the re-filtration operation was repeated twice at 10 minute intervals to obtain a third batch of compound XVa (0.134 g; 2.7%). The total yield of compound XVa was 4.27 g (86%).

A mixture of 4.20 g (4.22 mmol) of compound XVa, 1 ml of anisole and 40 ml of TFA was stirred for 1.5 hours at room temperature. The mixture was evaporated under reduced pressure at a temperature below 20 ° C and the dark residue was triturated with 300 ml of isopropyl ether to precipitate 3.50 g of the TFA salt, which was collected by filtration and dried under reduced pressure. The TFA salt was dissolved in 50 ml of methanol, treated with a small amount of charcoal and filtered. The filtrate was concentrated under reduced pressure and 15 ml of a 1M solution of sodium 2-ethyl hexanoate 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 compound as a crude product (estimated purity 50%), which was collected by filtration, washed with a small amount of ethyl acetate and dried. The ratio of δ-isomer to Δ-isomer of the crude product was 1: 4 (HPLC, Lichrosorb RP-18; mobile phase, 1 / 100M ammonium phosphate buffer, pH 7 - CH 2 OH, 90:10; Retention time, Δ²isomer 6 ′ 54, Δ²-isomer 8 '29).

The crude product (2.36 g) was dissolved in a small amount of water and purified using HPLC (Waters Associates, System 500, PrepPAK 500 / C g; mobile phase, 7% methanol). The HPLC eluate containing the product was concentrated under reduced pressure at a temperature below 35 ° C and the concentrate was freeze-dried to obtain the title compound. Yield 959 mg (42%, 15 based on the iodide Vila). Amorphous powder. Estimated purity 80% (by HPLC). The ratio of A 2 isomer to isomer was 1:17. The compound was gradually decomposed at a temperature above 160 ° C.

453 092