DE1445685A1 - Antibiotically active cephaloscopic acid derivatives and processes for their preparation - Google Patents

Description

Dr.-tng.HANS RUSCHKE
Dipl.-Ing. HEiNZ AGULAR

PATENTANWÄLTE Postal checking account: _

, West 74 M Monks 68277 "· *

Bank account: 00

HmcW II. tadwtri · Dresdner Bank _ω Diit »83 some ^ *" ^ Berti «38 Dep.-Kassel LeopotdttraB · LD Taplüzw SbaS · «Account 58615.-f.

"" "^ ZÜIV, Tetegram address: ^.

£ ÜL square monks "**

£ 11 Lilly and Company, Indianapolis, Indiana, P.St.A.

intibiotlsoh effective cephalosporanic acid derivatives and Process for their manufacture

The invention relates to antibiotic substances of the general formula

) ^ ₁

% ² s

I 2 / \

- (CHo) _n -C-IiH-CH-HC CH ₉

Il I

0 »C - Έ ■ C-CH ₀ -OC-OH

C ^u

COOH

in which R denotes a phenyl, chlorophenyl or furyl radical f in which m and η - when R is a phenyl radical - denote 0 or 1 »where the sum of m plus η is not greater than t

is, or - if R is a chlorophenyl radical or furyl radical -

t 2
0 means; and in which R and R are hydrogen atoms or methyl

809811/1090

groups are j and a pharmaceutically suitable salt this connection.

The invention also relates to a process for the preparation of an antibiotic cephalosporin compound of the above general formula, wherein a compound having the bicyclic ring structure of cephalosporin C _{9 is} given the formula

S \

H ₀ F-CH-CH CH ₀

III

O = C-Ii C-CH ₀ -OC-CH,

C.
1
COOH

having, acylated with an acylating agent which min at least one functional radical of the general formula

1*

E ¹

has, wherein R, m and η have the meaning given above

1 * 2. *
and R and R are hydrogen atoms or methyl groups or customary protective groups, such as carbobenzoxy groups,

1 * 2 *
where one of the radicals R and R is such a protective group if the amino group in the product is primary} whereby any protective group present is split off hydrogenolytically

8098T1 / t090

and optionally converting the product obtained into a pharmaceutical converts suitable salt.

The novel compounds of the invention are related to cephalosporin G in that they contain the 5,6-dihydro-2H-1,3-thiazine ring with the fused-on β-lactam ring in the 2,3-position, which is characteristic of cephalosporin C. is. In contrast to the cephalosporin C but which has at the 7-position, the ^5-amino-t IT -aäipamylgruppe ^l, compounds of the invention in the 7-position are characterized by an acylamino group, an amino group in the α- or ß-position, wearing. Moreover, in contrast to cephalosporin C, which has relatively little antibacterial activity, the compounds of the present invention are highly effective antibacterial agents which support the growth of numerous types

st of microorganisms in the various environments too

to inhibit.

You at the penicillins with which the invention Compounds to some extent related can be made pharmaceutically by combining them with non-toxic acceptable cations, anions, alcohol residues, ammonia and amines numerous salts, esters, amides and the like. Derivatives can be used. These derivatives are considered full-fledged Consider equivalents of the compounds described un4 are therefore also within the scope of the invention.

Some types of cationic «SaI-

8098 1 1/1090

called zen, which are made from the compounds that contain the cephalosporin C core: water-soluble Salts such as the sodium, potassium, lithium, ammonium and substituted ammonium salts, as well as less water-soluble ones Salts such as calcium, barium, procaine, quinine and dibenzylethylenediamine salts. Those connections which contain the Cephalosporin-C core, do not form cationic, but anionic salts, i.e. addition salts with strong acids such as hydrochloric acid, hydrobromic acid, Phosphoric acid, sulfuric acid and similar acids.

Cephalosporin C can be obtained by culturing a cephalosporin C producing organism in a suitable Hearing medium can be prepared as described in the British patent 810 196 is described.

The cephalosporin core can easily be made from cephalosporin C obtained by adding the 5 * -amino-] f'-adipamy! Side chain between the amide nitrogen and the amide carbonyl group splits. 7-Aminocephalosporanic acid can thus be prepared according to the process described in Belgian patent specification 593,777 produced by prolonged digestion of clay Cephalosporin C with a mineral acid in the absence of light will.

The compounds according to the invention are obtained by acylation of the 7-amino group of the cephaloeporane ^H nuclei - as indicated above - by means of the customary acylation process and using the various types

ι η Q η

manufactured by known sealing agents which have a Have composition that of the desired side chain is equivalent to.

A convenient acylating agent is the corresponding amino-substituted acyl chloride or bromide in which the amino group (if it is a primary amino group) is protected in the usual manner with a carbobenzoxy, carboallyloxy, tert-butoxycarbonyl, trityl radical or the like is. Me acylation is carried out in water or a suitable organic solvent, preferably under substantially neutral conditions, and preferably at or slightly below tree temperature, that is between the freezing point of the reaction mixture and about 2O ⁰ C. In a typical method, the 7-aminocephalosporanic acid or one of its derivatives dissolved together with a sufficient amount of Hatriumbicarbonat or other suitable alkaline compound to accelerate the solution in an aqueous 50 vol .- ^ acetone, wherein the concentration of 7-aminocephalosporanic acid about 1-4 wt -. $> is. The solution is cooled to about 0 to 5 ° C. and, while stirring and cooling, a solution of the acylating agent with a protected amino group is added in about 20% excess. If the pH value of the Gemiso does not remain constant, it can be adjusted by adding sodium bicarbonate or by introducing carbon dioxide

809811/1090

be kept in the neutral range. After the addition. of the aylating agent, the reaction mixture is stirred further and allowed to warm to room temperature. That . _{:: The} reaction product is then acidified to about pH = 2 with hydrochloric acid and extracted with an organic solvent such as ethyl acetate. The ithylacetate extract is adjusted to about pH 5.5 with sodium, potassium or ammonium hydroxide or another base containing the desired cation and extracted with water. The aqueous solution is separated off and evaporated to dryness. The residue is taken up in the smallest possible amount of warm methanol and the desired product is deposited by cooling and / or evaporation, if necessary with the addition of ethanol as a precipitant. The crystalline product obtained is filtered off, washed with acetone and dried.

Before or after the crystallization, the product or crude product can be treated in the usual way in order to remove the protective group (if used) from the amino group, which is suitably achieved by hydrogenation under mild conditions in the presence of a palladium catalyst or by brief treatment under weakly acidic conditions at elevated temperature (for example 30- to 2-minute treatment with dilute acetic acid at 50 100 ⁰ C) happens. The desired end product is thereby obtained.

You acylation can also be done with the corresponding amino

809811/1090

carboxylic acid itself in connection with an equimolar amount of a carbodiimide, vie ^, N'-diisopropylcarbodiimide, H, N * -dicyclohexylcarbodiimide, N, H ^f -Bis (p-dimethylaminophenyl) ■ carbodiimide, N-ethyl-N ¹ - (4 " -äthylmorpholinyl) -earbodi: ijiid or the like., The acylation takes place in such cases at normal temperature.

Optionally, the aminocarboxylic acid can be converted into the corresponding acid anhydride, the azide or an activated one Esters, any of these derivatives can be used to achieve the desired acylation. Other known means need not be listed here will.

In most cases the acylating agent contains one or more asymmetric carbon atoms and therefore occurs in optically active forms. In the ordinary In chemical synthesis, these compounds are usually in the form of the racemate, i.e. an equimolar mixture of the optical antipode showing no optical rotation. Will be the separated optical isomers if desired, the acylating agent can be cleaved in the usual way, e.g. by reacting the free acid with cinchonine, strychnine, brucine and the like, fractional crystallization of the distereoisomeric salts and separated Acidification of the solid phase and the liquid phase to obtain the to release optical antipodes. The free acids obtained in this way can be used as such for acylation

80981 1/1090.

can be used, preferably in conjunction with a carbodiimide, or can be converted into the corresponding acid halide or a mixed anhydride by conventional methods be converted, whereby of course extreme conditions must be avoided so that no Eacemieierung occurs.

In the literature, many of the acylating agents used in the present invention are described along with procedures their preparation has been described and a number of them are commercially available. All connections can be easily prepared by well-known methods.

The invention is further illustrated by the following examples, which, however, are not intended as a limitation of the Area of the invention are to be understood.

example 1
• T-CD-α-aminophenylacetaainoJ-eephalosporanic acid

_dj 1 -Phenylglycine is used in the usual way Reaction with cinchonine, fractional crystallization of the diastereoisomeric salts obtained and acidification to obtain the To obtain phenylglyein antipodes, cleaved.

D-phenylglycine produced in this way becomes reacted in the usual way with Garbobenzoxychlorid to to obtain the H-öarbobenzoxy-D-phenylglycine.

0.60 g of N-carbobenzoxy-B-phenylglyein are in 10 ecu of dry! Tetrahydrofuran dissolved. The solution will be

Chilled in an ice / salt bath and stirring inside

8 0 9 8 11/109 0

0.29 ecm of triethylamine was added for 10 minutes, followed Clay 0.29 ecm isobutyl oil ester, followed by more 10 minutes "was stirred at -5 ° C. During this time, 0.57 g of 7-aminocephalosporanic acid and 0.29 ecm of triethylamine dissolved in 5 ecm tetrahydrofuran and 5 ecm water, and the The solution is centrifuged to remove a dark colored sludge. The clarified solution is in an ice bath cooled and slowly added to the heating mixture, whereby another half hour in the ice bath and another hour at tree temperature. The reaction mixture is a homogeneous solution with a pH of about 6. The solution turns into a semi-solid residue evaporated * 35 ecm of water and a few drops of triethylamine are added to the residue to adjust the pH increase to 8. The resulting aqueous solution is successively with 50 and 35 ccm portions of Ethyl acetate extracted, the pH being adjusted to 2 with hydrochloric acid for each extraction. The extracts were combined, filtered, dried over sodium sulfate, the solvent stripped off and the residue dried in vacuo, 7- (H-carbobenzoxy-D-α-aminophenylacetamino) -cephalosporic acid is obtained in the form of a yellow-white, amorphous solid mass with a yield of 1.10 g.

1.0 g of this substance are dissolved in 95 ml of ethyl alcohol at 150 ecm. 1.0 g are added to the solution given a 5 igen palladium / carbon catalyst, and that

1/1090

Mixture is carried out at room temperature and normal pressure. Introducing hydrogen over a period of 3 hours, hydrogenated with stirring. The hydrogenation mixture is filtered. The solid phase consisting of the catalyst and the desired product is suspended in ethyl acetate and water and adjusted to pH = 2 with hydrochloric acid. The suspension is filtered to remove the catalyst. The aqueous phase is separated off from the filtrate and evaporated down by evaporation, with the desired 7- (D ~ α-aminophenylaeetamino) -cephalosporanic acid is obtained.

Example 2

7- (d ^ t-a-Amino phenylac et amino) cephalospo ranoic acid

To a solution of 1.4-8 g of N, IST'-dicyclohexylcarbodiimide in 100 ecm of tetrahydrofuran, 2.05 g of d.1- ÜT-carbclsenzoxy-a-aminophenylacetic acid are added, followed by a solution of 2 g of 7-aminocephalosporanic acid in 7 »2 ecm aqueous 1 normal sodium hydroxide solution, and the mixture is stirred overnight at room temperature. The tetrahydrofuran and the water are drawn off in vacuo. The oil obtained is dissolved in water, filtered and extracted with ethyl acetate at pH = 2, hydrochloric acid being used to adjust the pH. The ethyl acetate extract is separated off and the solvent is stripped off in vacuo. The residue is washed with ether, redissolved in ethyl acetate and extracted with water at pH 6.5,

809811/1090

aqueous potassium hydroxide solution being used to adjust the pH. The extract obtained is evaporated to dryness i.V. The oil obtained is redissolved in Methajbl and through Gradual evaporation of the methanol and simultaneous addition of ethanol as amorphous solid hate precipitated · It will be 278 mg of 7- (α-carbobensoxyamino-α-phenylacetamino) -cephalosporanic acid obtained in the form of the potassium salts of the diastereoisomers.

The material thus obtained is subjected to selective hydrogenolysis as in Example 1 in order to remove the carbobenzoxy-Sokutsgruppe from the α-amino group of the side chain jsu, whereby 200 mg of 7- ( i.e. 1 -α-aminophenylaoetamino) -oephalosporanic acid are obtained as the end product.

Example 5

7- ( d.1- g-aminophenylaoetamino) -cephalosporanic acid

To a Gemisoh tone 230 com dioxane and 15 oam acetone are added 2.1 com triethylamine followed by 4.25 gd ^ Ji-li-Oarbobenz-oxy-a-aminophenylacetic acid. The solution obtained is ^{cooled to 0} ° C. and, over the course of 15 minutes, a solution of 2.04 g of isobutyl chloroformate in 15 cm of acetone is added dropwise with stirring. Each addition is complete, the mixture is 10 minutes

80-981 1/1090

left to stand at ⁰ ° C. A solution of 4.1 g of 7-aminocephalosporanic acid and 2.1 com triethylamine in 15 com water is then added. This mixture is left to stand at room temperature for 2 hours. It is then diluted with 300 ecm of water and washed twice with ether. The washed solution is acidified to pH * 2 with 1-noreal hydrochloric acid and extracted with 300 ecm ethyl acetate. The ethyl acetate extract is washed once with water and then extracted at pH 6.5 with 100 com of water, the pH being adjusted with 1 normal potassium hydroxide solution. The aqueous extract is separated off and evaporated to dryness iY. The resulting crystalline residue is recrystallized from methanol, 2.5 g of 7 -d.1- H-carbobenzoxy-a-aminophenylacetamino) -cephaiosporanic acid being obtained in the form of the potassium salt.

The intermediate product thus obtained is selectively hydrogenated as indicated in Example 1, whereby 7- ( i.e. 1- e-iminophenylaoetamino) -cephalosporanic acid is obtained as the desired end product.

Example 4

7- (L-a-imino-fi-phenylprepionamido} -cephalo sporanic acid

To produce K-carbobenzoxy-L-S-phenylalanine, L-ß-phenylalanine is reacted with carbobenzoxyehleride in the usual way.

809811/1090

3.0 g of the product obtained are dissolved in a mixture of 3 coat Diöxan and 15 ecm acetone. The solution MfXtU "stirred, cooled to ⁰ -5 C and treated with 2.1 cc of anhydrous TriäthyTamin. A solution of 2.04 g of isobutyl chloroformate in 15 ee" of acetone is then added over about 15 minutes. The mixture is stirred for another 10 minutes A cold solution of 4.1 g of 7-aminocephaloboranic acid and 2.10 ecm of triethylamine in water is then added all at once, the mixture is warmed to tree temperature and left to stand for 2 hours and then evaporated practically to dryness iV. The residue is taken from methanol and recrystallized ethanol, 1.63 g of 7- (N-carbobenzoxy-La-amino-β-phenylpropionamido) -eephalosporanic acid being obtained The compound has an antibiotic activity corresponding to 33 units of penicillin G per mg.

100 mg of the material obtained in this way are dissolved in 10 cos water. A suspension of 1 g of 5% palladium / barium sulfate catalyst in 10 ecm of water is added to the solution, and the mixture is stirred overnight (about 16 hours) in the presence of hydrogen at an atmospheric pressure. The suspension is filtered. The solid phase, which contains the catalyst and the desired product, is suspended in ethyl acetate and water and acidified to pH = »2 with hydrochloric acid. The mixture will

& 09811/1090

-H-

then filtered and the filtrate separated into the two layers. The aqueous phase is brought to 100 ecm. The biological test reveals an antibiotic Effectiveness corresponding to 21 units of penicillin G per ecm. The paper chromatography of the solution with aqueous 70% n-propanol as the mobile phase provides a heck of a problem. _. , biologically active material that is different from the starting material. This stain is 7- (I-a-amino-ß-phenylpropionamido) -cephalosporanic acid.

809811/1090

Claims (1)

Claims 1. Intibiotically effective cephalosporanic acid derivative of general formula R- (CH ₀ ) -CH - (CH ₀ ) -C-HH-CH-HC CH ₀ cm there ι · ι c. 0 »C-H C-CH ₀ -OC COOH in which R is a phenyl, chlorophenyl or furyl radical} in which m and η when R is a phenyl radical - denote 0 or 1, the sum of m plus η not being greater than 1 is, or - if R is a chlorophenyl radical or furyl radical - 1 2
0 means; and in which R and R are hydrogen atoms or methyl groups} and a pharmaceutically acceptable salt of this compound. 2. A process for the preparation of an antibiotically active cephalosporanic acid derivative of the general formula 1 according to the preceding claim 1, characterized in that 7-aminocephalosporanic acid or one of its salts is acylated in a manner known per se with an acylating agent which contains at least one constitutional radical of the all- 809811/1090 Documents (Art 7, Section I, Paragraph 2, No. I, Clause 3 of the Amendment Act of 4.9. common formula 1* E ¹ 2 *
E- (CH ₂ ) _m -CH- (CH ₂ ) _η - auf ei st, where E, m and η have the meaning given above 1 * 2 *
and E and E denote hydrogen atoms or methyl groups or customary protecting groups such as carbobenzoxy groups, one of E and E being such a protecting group when the amino group in the product is primary; that any protective group present is split off hydrogenolytically and that the product obtained is optionally converted into a pharmaceutically suitable salt. 80.98 1 1/1 090