DE2344451C2 - 7 - [β-Aminoacylamino] -3-substituted-3-cephem-4-carboxylic acid derivatives, processes for their preparation and pharmaceutical compositions containing them - Google Patents

Description

(a) a 7-amino-3-substituted-3-cephem-4-carboxylic acid the general formula

H ₂ N-, (

yy-

CH ₂ -SR ₃
COOM

(Π)

wherein R3 and M are those given in claim Have meanings

with a jJ-aminocarboxylic acid derivative of general formula

R ₁

HC-CH ₂ -COOH
X

(HI)

wherein Ri has the meaning given in claim 1 and X is a protected amino group means or

acylated with a reactive derivative thereof and then if necessary the Amino protecting group removed; or

a 7-acylated aminocephalosporanic acid der general formula

I J-CH ₂ -OCOCHj

COOM

(IV)

in which R- and M have the meanings given above and X 'denotes an amino group or a protected amino group,
with a nucleophile of general form!

R, SH (V)

wherein Rj has the meaning given in claim 1. implements and then if necessary the amino protection group removed.

The invention relates to new 7 [/? - Aminoacylamino] -3-substituted-S-cephem ^ -carboxylic acid derivatives. which have valuable antibacterial activity. Process / u of their production as well as containing them pharmaceutical agents.

Various cephalosporins or cephemcarboxylic acid derivatives are already known which have an antibacterial Have activity, for example in "the German Publenlegungsscnrifteri 16 70 600 And 18600 antibiotically effective cephalospofin compounds with an Aminöäcylamidögruppe in the 7-position of the cephalosporin nucleus or, antibiotisdh effective 3'ThiadiazOlyllhio · and 3-Telfäzolylthiöcephälösporine known that the growth of both gram-neutral and gram-positive bacteria inhibit.

But on the one hand the anti-bacterial effectiveness of the previously known cephalosporins is certain Is subject to restrictions, on the other hand, the occurrence of mutation, compared to the known cephalosporins-resistant microorganisms

ω human strains cannot be excluded, was The task of the invention is the search for new antibacterials * rially effective compounds with an even higher antibaktefieilett activity, further the possibility offer, the development of resistant microorganisms ^ to counteract men.

It has now been found that this object can be achieved according to the invention by new ones [SAili

Acid derivatives of the general given below Formula (I), which have a significantly higher antibacterial activity than relevant commercial products (See the comparative example below) and by the method for their preparation.

The process according to the invention for the preparation of the compounds I is characterized in that one in a manner known per se

(a) a Z-amino-S-substituted O-cephem ^ carboxylic acid the general formula

m

COOM

wherein R ₃ and M have the meanings given,

with a / 9-aminocarboxylic acid derivative of the general formula

HC-CH ₂ -COOH

(IFl

wherein R, has the meaning given and X is a protected amino group, or acylated with a reactive derivative thereof and then if necessary the amino protecting group removed; or

a 7-acylated aminocephalosporanic acid of the general formula

H -C-CH ₂ CONH-, / '

X '

"J-CH ₂ -OCOCH ₃ (IV)

COOM

wherein Ri and M have the meanings given and X 'denotes an amino group or a protected amino group, with a nucleus of the general formula

R ₃ SH V

wherein R _{3 has} the specified Bedeatunr, reacts and then if necessary removed the amino protective group.

The 7 - [/? - Aminoacylamino] -3-substituted-3-cephem-4-carboxylic acid derivatives according to the invention the general formula

R ₁

HC-CH ₂ CONH-, 1 ^

NH,

where mean:

CH ₂ -SR ₃

(D

45

50

Ri is a phenyl or thienyl group, R _{3 is} a 3-methyl-1,2,4-thiadiazol-5-yl-, 5-methyl-1,3,4-thiadiazolyl-, 1-methyl-1H-tetrazolyl- or

2-methyl-2Htetrazolyl group and M is a hydrogen atom or a non-toxic, pharmaceutically acceptable cation,

60

Have a high antibacterial activity and effectively prevent: that; Growth of a large number of microorganisms, including the gram-positive and gram-negative bacteria, with regard to their antibacterial effectiveness, they are clearly superior to the relevant hand products, as the results of comparative experiments given in the comparative example below demonstrate. According to a further aspect, the invention relates in that they contain further active compound ^c al pharmaceutical agents which are characterized by a compound of the above general formula (I) in admixture with a pharmaceutically acceptable carrier or diluent suitable for oral or parenteral administration. The carriers or diluents are organic or inorganic in nature.

The pharmaceutical agents according to the invention can be in solid form, e.g. B. in the form of capsules, Tablets or dragees, or in liquid form, e.g. B. in the form of solutions, suspensions or emulsions, are present. If desired, the pharmaceutical agents according to the invention can additionally contain further ones Contain auxiliary substances, such as. B. stabilizers, wetting agents or emulsifiers, buffers or others on additives commonly used in this field.

Even if the dosage of the antibacterially active compounds according to the invention is dependent depends on the age and health of the patient, has become an average Single dose of about 100 mg. 250 mg and 500 mg of compound as effective for treating disease proven to be caused by a bacterial infection be evoked. In general, amounts can be between 10 mg and about 1000 mg or even more higher amounts are administered.

The term "pharmaceutically acceptable cation" as used herein means essentially non-toxic cations, such as those of the alkali metals, z. B. Sodium and Potassium.

The term "protected amino group" represented by the symbol X in formula (MI) denotes one Amino solid with a protecting group, such as benzyloxycarbohyl,

4 ^j nitroben2yloxycarbonyl,

4-bromobenzyloxycarbonyI,

4 ⁱ MethoxybenzyIoxycarbonyi, 3,4-Dimethoxybenzyloxycarbonyl, 4- (Phenyläzo) benzyloxycäfbönyl,

4- (4-methoxyphenylazo) benzyIoxycarbonyl,

t-butoxycarbonyl, t-pentyloxyearbonyl,

Isopropoxycarbonyl,

Diphenylmethoxycarbonyl,

2-pyridyimethoxycarbonyl,

^^ - Trichlorethoxycarbonyl,

2,2,2-tribromoethoxycarbonyl,

1 -cyclopropylethoxycarbonyl,

3-iodo-propoxycarbonyl,

2-Fun'uryIoxycarbonyl,

1 - adamantyloxycarbonyl,

S-quinolyloxycarbonyl.trifluoroacetyl,

Trityl, 2-nitrophenylthio,

2,4-DinitrophenyIthio, 2-Hydroxybenzylidene,

2-hydroxy-5-chlorobenzylidene,

2-hydroxy-1-napthylmethyien,

S-Hydroxy-'t-pyridylmethylene,

l-lVtethoxycarbonyI-2-propylidene,

1-ethoxy-carbonyl-2-propylene,

S-ethoxycarbonyl ^ -butylidene,

1 - acetyl-2-propylidene,

1-propionyl-2-propylidene,

1 -Benzoyl-2-propylidene,

13-bis (ethoxycarbonyl) -2-propylides,

1- (N-M ': thylcarbamüyI) -2-propyIiden,

1 - (N ₁ N-dimethylcarbamoyl) -2-propylidene,

1- [N- (2 methoxyphenyl) carbamoyl] -2-picpyüder.,

1 [N- (4 methoxyphenyl) carbamoyl] -2-propylidene,

] - (N-PbenylcarbamoyI) -2-propyIiden,

2-Äthoj-ycarbonylcyclopentyliden,

2-EthoxycarbonylcycIohexyIiden,

2-acetylcyciohexylidene and

SJ-dimethyl-S-oxocyclohexylidene.
Further examples of protected amino are the acid salts of the free amino group such as hydrochloride, hypobromide and hydroiodide.

The acylation reaction can be carried out in a customary manner be carried out, e.g. B. in a solvent such as acetone, dioxane, acetonitrile, chloroform, ethylene chloride, Tetrahydrofuran. Ethyl acetate. Dimethylformamide, pyridine or water, or any other suitable organic solvents. The reaction can preferably be carried out with cooling or at room temperature be performed.

When the / J-amino carboxylic acid derivative of the formula (III) is used in the form of the free acid or the salt, the acylation reaction in the presence of a Condensation agents such as N.N'-dicyclohexylcarbodiimide or thionyl chloride. Suitable reactive derivatives of the aminocarboxylic acid derivatives of the formula (111) are carboxylic acid halides, active amides, active esters. Acid anhydrides or mixed anhydrides with other carboxylic acids.

The acylation reaction can also be carried out after the 7-amino-3-substituted-3-cephem-4-carboxylic acid of formula (II) with a silylating agent. such as chlorotnmethylsilane. Llis- [N, N-trimethylilyljacetamide or N trimethylsilylacetamide.

_r "; After the acylation, the protective group from the amino radical and the silyl group are removed, if necessary.

The reaction for removing the protective group from the amino radical can be carried out in a known manner Will, e.g. B. by treating the acylation product with acids such as formic acid or trifluoroacetic acid ^ acid, or by catalytic reduction, depending on the protective group to be removed.

The split-off reaction using Acids can be used, for example, for those compounds that are used as a protecting group, for. B. benzyloxycarbonyl, substituted benzyloxycarbonyl, alkoxycarbonyl, substituted alkoxycarbonyl, aralkoxy carbonyl, Adamantyloxycarbonyl, trityi, substituted phenylthio, substituted aralkylidene, substituted alkylidene or ίο have substituted cycloalkylidene. The catalytic Reduction as a cleavage reaction can be applied to compounds that act as protecting groups z. B. Benzyloxycarbonyl, substituted benzyloxycarbonyl or 2-Pyndylmethoxycarbijnyl, π If the protecting group is a trifluoroacetyl group, it can be removed by treating the acylation product removed with water. When the protecting group is haloalkoxycarbonyl- or 8-quinolyl-oxycarbonyl it can be removed by treating the acylation product mk copper or zinc.

Removal of the Silylgr. ^ Pe, which is usually

easily through the aftertreatment of the acylation reaction product Removed in the acylation reaction can be done by treating the acylation reaction product done with water.

LMe reactions to remove the protective groups on the amino radical and the silyl group can be carried out without Isolate the acylation reaction product from the reaction medium.

The implementation of a 7-acylated aminocephalosporanic acid of formula (IV) with a nucleophile of formula (V) can in a solvent such as water, Acetone, chloroform. Nitrobenzene, dimethylformamide. Methanol, ethanol. Dimethyl sulfoxide or any other suitable organic solvents. The reaction can take place at room temperature or with heating. When the nucleophile of formula (V) is in the form of the free thiol is used, the reaction is preferably carried out in the presence of a base, e.g. B. of an Aikalime tallhydroxids, an alkali metal carbonate, an alkali metal bicarbonate or a trialkylamine. If the 7-acyiated aminocephalosporanic acid of formula (IV). in which X 'is a protected amino group. is used, the desired Ve-compounds of the formula (I) can be obtained by the Protective group on the amino group removed in the usual manner as set out above.

It is generally known that the majority of the compounds of formula (I) has an asymmetric carbon atom and therefore optically active in two isomeric forms, the D and L-Fo ^r men, is present. The invention includes both the D and du. "L, forms and the DL mixtures of these compounds

gene.

The reaction components used for the process according to the invention are commercially available obtainable or can by customary, generally known processes or by analogy processes, the are applicable to the preparation of such reaction components.

According to -dem. The method of the invention can be a precipitate formed during the reaction are _t enifcrnt from the reaction mixture by methods such as are commonly used for this purpose, and the reaction products obtained can the purification methods routinely used are subjected z, B, recrystallization from

a suitable solvent or a mixture of solvents.

The compounds of the formula (I) can by customary methods of salt formation from the acids in their pharmaceutically acceptable, essentially non-toxic salts are converted, e.g. B. by Reaction with an alkali metal hydroxide, an alkali metal bicarbonate, an alkali metal carbonate, or an organic base. The sodium salts are preferred. The preferred method of manufacture of salts consists in dissolving the acid in a solvent in which the salt is insoluble, and that a solution of the salt-forming compound is then added to the base. The salt falls as Precipitation from the reaction mixture.

The invention is illustrated by the following examples.

example 1

Pivaloyl chloride (4.9 g) was added all at once to a dry acetone solution (150 ml) of 3- (N-tert-butoxycarbonylamino) -3-phenyl-DL-propionic acid (10.8 g) and triethylamine (4.1 g) with stirring - 10 to -15 ° C given. The resulting mixture was for 1 hour at -10 to - 15 ^r stirred C. Then at -30 to -40 ° C with stirring a mixture of

7-Amino-3- (5-methyl-1 3,4-thiadiazoI-2-yl) thiomethyl-3-cephem-4-carboxylic acid (14.0 g), triethylamine (12.2 g), water (40 ml) and acetone (160 ml). The mixture was left at -30 to -40 ° C for 1.5 hours and then at room temperature for 1.5 hours touched. After the acetone had been removed from the reaction mixture under reduced pressure, UuHe the residue obtained with ethyl acetate gev, a, then The water layer was separated, with JO% hydrochloric acid adjusted to about pH 1 and with Ethyl acetate (1000 ml) extracted. The organic layer was washed with water and an aqueous solution of sodium chloride, dried over magnesium sulfate and treated with activated charcoal. The organic solution was then reduced to a reduced pressure Volume of about 100 ml concentrated, and the concentrate was filtered, whereby

7- [3- (N-tert-ButoxycarbonyIamino) -3-phenyl-DL-propionamido] -3- (5-methyl-13.4-thiadiazol 2-yl) thiomethyl-3-cephem-4-carboxylic acid (1.48 g) ,
Mp 150-155 ° C (dec.).

The mother liquor was also concentrated under reduced pressure, and the deposited precipitate was filtered when only a little ethyl acetate remained. This gave 13 g of the same product obtain.

IR (Nujol)

3350.1775,1710,1685.1660.1525 cm- '
UV (phosphate buffer, pH 6.4)

A _rat 272mu, E = 208

The above-mentioned 7- [3- (N-tert-ButoxycarbonyIamino) -3-phenyl-DL-propionamido] -3- (5-methyl-1 _r 3,4-thiadiazoI-2-yI) -thiomethyl-3-cephem -4-carboxylic acid (1.0 g) was added to Seiger formic acid, and the mixture was ^{stirred at 40 °} C. for 3 hours. After the reaction mixture had been concentrated to a volume of about 5 ml under reduced pressure, water (about 2 ml ) and methyl isobutyl ketone: containing Amberlite LA-I (about 5 ml) was added to the residue, and the mixture was stirred at room temperature overnight. The precipitate: was filtered to give a pale-yellow powder from 7- (3-Airiino-3-phenyl-DL-propioriamido) -3- (5-methyl ^l l, J, 4-thiadiazo -2-y!) tniomelhyI-3-cephem-4-earbunic acid (170 mg) was obtained.

IR (Nujol)

3300.1760.1650.1590.1530 cm- ' to UV (phosphate pepper, pH 6.4)

Example 2

3- (N-lert.-Butoxycarbonylamino) -3- (2-thienyl) -DL-propionic acid (10.8 g) and triethylamine (4.04 g) were dissolved in dry acetone (200 ml) with stirring at room temperature. After cooling the solution to -10 to -15 ° C, pivaloyl chloride (4.82 g) was added to the solution all at once with stirring. The mixture was 1 hour at - 15 ° C and stirred then cooled to -40 to -5o C ⁰ - 10 to. A mixture of 7-amino-3- (5-methyl-1,3,4-lhiodiazol-2-yl) thiomethyl-3-cephem-4-carboxylic acid (13.76 g),

Triethylamine (12.0 g), water (40 ml) and acetone (160 ml) were added all at once. The resulting mixture was 1.5 hours at -40 to -40 ⁰ C and then 2 hours be '. Room temperature stirred. The acetone was removed from the reaction mixture under reduced pressure and the residue was washed with ethyl acetate. The aqueous layer was separated, adjusted to pH 1 with 10% hydrochloric acid and extracted with ethyl acetate. The extract was washed with water and an aqueous solution of sodium chloride, dried over magnesium sulfate, treated with activated charcoal, and then concentrated under reduced pressure. Ether was added to the concentrate and the mixture was stirred for 3 hours. The precipitate was filtered off, whereby 7- [3- (N-tert-

■ Ό Butoxycarbonylamino) -3- (2-thienyl) -DL-propionamido] -3- (5-methyl-13,4-thiadiazoF-2-yi) thiomethyl-3-cephem-4-carboxylic acid (15.1 g) mp 130-135 ° C (dec).

IR (Nujol)

3350,1780,1710,1690.1650 cm - ' UV (phosphate buffer, pH 6.4)

A _ma * 235πιμ, £ = 255

λ _5Α 272mu, £ = 199

The 7- [3- (N-tert-butoxycarbonylamine;.) ⁴ -3- (2-thienyl) -DL-propionamido] -3- (5-methyl-13,4-thiadiazoI-2-yl) - thiomethyl-3-cephem-4-carboxylic acid (10 g) was mixed with 99% formic acid (100 ml) under

κ ice cooling stirred for 2 hours The reaction mixture was then concentrated under reduced pressure to leave a pale brown pasty residue was obtained. The concentrate was successively washed with ether (100 ml), ethyl acetate (100 ml) and acetonitrile (100 ml). The precipitate was filtered a pale yellow powder (5.6 g) was obtained. The product thus obtained (43 g) was dissolved in water (approx 10 ml) dissolved Amberlite® LA-I (approx. 60 ml) was added to this. The mixture was at overnight Stirred room temperature The precipitate was filtered, digested with acetone and with acetone and with Ether washed, a pale yellow powder (133 g) 7- [3-Amino-3- (2-thienyI) -DL-propionamido] -3-

(5-methyI-1,3,4-thiadiazol-2-yl) thiomethy ['3-cephem4-ciarboxylic acid, 185-186 ° G (decomp.). After the mother liquor thus obtained was also concentrated under reduced pressure, was the concentrate washed with acetone and ether, whereby the same product (1.1 g) was obtained » Total yield; 2.43 g,

IR (Nujol)

3300,1780,1660,1630,1580,1520 crh- ' UV (phosphate buffer, pH 6.4)

A _m "232 πιμ, E = 286

λ, Α 273 mn, E = 237

Example 3

7- [3- (N-tert-ButoxycarbonyIamino) -3-phenyl-DL-propionamido] -3- (3-methyl-1,2,4-thiadiazol-5-yl) thio- methyiö-cepnem ^ -carboxylic acid (2, ög) was with

Formic acid reacted, with 7- (3-amino-3-phenyl-DL-propionamido) -3- (3-methyl-1,2,4-thiadiazoI-5-yl) -

thiomethyl-3-cephem-4-carboxylic acid (1.8 g), ^{mp 135 to 140 0} C (dec.) was obtained.

UV (phosphate buffer, pH 6.4)

Example 4

To a mixture of 7- [3- (N-tert-butoxycarbonyl _t äiiiino) -3-phenyl-DL-propionamido] cephalosporanic acid (2 g) and 5-methyl-1,3,4-thiadiazole 2-thiol (0.5 g) in; phosphate buffer (pH 6.4, 60 ml) was gradually added to sodium hydrogen carbonate (0.5 g). The J.'erhaltene solution was stirred for 6 hours at 60 ⁰ C, cooled and shaken with ether. The aqueous layer was separated, adjusted to pH 2 with 10% hydrochloric acid with cooling, and extracted with ethyl acetate. The organic layer was washed successively with water and a saturated aqueous common salt solution, dried over magnesium sulfate, and concentrated under reduced pressure. The residue obtained was digested with ether, a powder _Λ (0.85 g) of 7- [3- (N-tert-butoxycarbonylamino) -3-phenyl-DL-propionamido] -3- (5-methyl-13, 4-thiadiazoI-2-yl) -thiomethyl-3-cephem-4-carboxylic acid was obtained. This powder was dissolved in ethyl acetate. Then most of the ethyl acetate was removed under reduced pressure. After cooling, the precipitate was filtered off and dried to give the desired compound (0.55 g), mp 148-154 ° C.

IR (NuJoI)

3350.1775.1710.1685.1660.1525 cm- · UV (phosphate buffer pH 6.4)

propionamido] -3- [1 -methyl-1-H -letrazol-5-yl] thiomethyl-3-cepheffi-4-carbofic acid Was received.

Example 6

S-fN-tert'Butoxycarbonylaminoj-S - ^ - thieny ^ DL-propionyl chloride and 7-Ariiino-3- (1 -methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid Were treated as starting materials in a manner analogous to that described in the above examples, with 7- [3-Αιτίίηο ^ (2-thienyl) -DL-propionarriido] -3- (1-methyl-IH-tetfazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid was obtained.

Comparative example

To demonstrate the superior antibacterial effectiveness of the compounds according to the invention, the comparative tests described below were carried out, the results of which are summarized in the tables below.
It became antibacterial effectiveness

The further processing of the desired end product takes place analogously to Example 1.

Example 5

3- [N-tert -Butoxycarbonylamino] -3-phenyl-DL-propionyl chloride and 7-amino-3- [1-methyl-1H-tetrazoI-5-yl] thiomethyl-3-cephem-4-carboxylic acid the starting materials were treated in a manner analogous to the above examples, with 7- [3-amino-3-phenyI-DL-der

compared the following compounds:

Connection A:

7- (3-Amino-3-phenyl-DL-propionamido) -3- (3-methyll, 2,4-thiadiazol-5-yl) thiomethyl-3-

cephem-4-carboxylic acid (described in the above

Example 3);
Connection B:

7- [3-Amino-3- (2-thienyl) -DL-propionamido] -3- (1 -methyl-1H-tetrazol-5-yl) thiomethyl-3-

cephem-4-carboxylic acid (described in the above

Example 6);
Connection C:

7-D-et-PhenylglycylcephaIosporanic acid (trade name "Cefaloglycine", described in Example 1 of

DE-OS 16 70 600);
Connection D:

7-D-Mande! Amido-3- (l-methyl-1,2,3,4-tetra-

zol-5-thiomethyl) -d ³ -cephem-4-carboxylic acid
(Trade name »Cefamandol«, described in Bp · -

game 4 of DE-OS 20 18 600);
Connection E:

Cefalexin.

45 Test Procedures

In vitro bacterial activity was determined using the two-fold agar plate dilution method.

A loop filling of an overnight culture of each of the microorganism test strains given in the tables below in trypticase soy broth (10 ⁸ viable cells per ml) was streaked on a heart infusion agar (HI agar) * the graded one

Concentrations of each compound tested and it was the minimum inhibitory concentration (MIC), expressed in μg / ml, after 20 hours Incubation at 37 ° C determined

Test results
Table (A)

Microorganism test strain

Investigated connection
ACDE

Staphylococcus
aureus 213

0.78 25 1.56 6.25

12th

continuation

Microphanism Test Strain

Investigated connection ACDE

Stäphylococcüs aureus 226

Table (B)

0.78 12.5 1.56 6, ^ 5

Microorganism test samples

Investigated connection BCDE

E. GoIi NIHJ JG-2 6.25 25 12.5

Klebsieila 3.13 200 50 6.25

pneumoniae 418

From the above tables it can be seen that the 20 had bacterial activity similar to that of the According to the invention, compounds A and B are used in the case of the known comparative products G, D and E The aforementioned microorganism test strains were superior to at least 100%.

Claims (5)

Patent claims: 1. 7-fJ? -Aminoacyiamino] -3-substituted-3-cephem-4-carboxylic acid derivatives, characterized by the general formula HC-CH ₂ CONH COOM (I) where mean: Ri is a phenyl or thienyl group, S-methyl-l ^ -thiadiazolyi-, 25th 30th 2-MethyI-2H-tetrazoIylgruppe and
M is a hydrogen atom or a non-toxic, pharmaceutically acceptable cation. 2. 7- [3-Amino-3-phenyI-DL-propionamido] -3- (3-methyl-1,2,4-thiadiazoI-5-yl) thiomethyl-3-cephem-4-carboxylic acid. 3. 7- [3-Amino-3- (2-thienyl) -DL-propionamido] -3- (1 -methyl-1H-tetrazoI-5-yl) thiomethyl-3-cephem-4-carboxylic acid. 4. Pharmaceutical agent, characterized in that it is a compound according to the active ingredient any one of claims 1 to 3 in admixture with a pharmaceutically acceptable carrier or diluent contains R, H -C-CH ₃ CONH I.
χ 5. Process for the preparation of 7 - [/? - Aminoacylamino] -3-substituted-3-cephem-4-carboxylic acid derivatives according to claims 1 to 3, characterized in that one in a conventional manner