DE2144457C3 - α-Aminobenzylpenicillin esters, processes for their preparation and pharmaceuticals containing them - Google Patents

Description

reacted, where A, R and R ^{1 have} the above meaning and -CO-Y ¹ and X ¹ -NH- groups which can react with one another to form a - CO - NH - group, and then optionally the group A in converts an amino group and / or reacts the compound obtained with a pharmaceutically acceptable acid to form the acid addition salt.

5. Medicaments consisting of one or more compounds according to claim 1 and usual Carriers.

iX-Amionobenzylpenicillins are known to be antibiotics with a broad spectrum of activity. However, their clinical significance is not limited to their pharmacological significance Properties, but also their absorbability when administered orally, their distribution in the Tissues, their transformation in the body and their speed of excretion are important. Numerous, for their good pharmacological properties (X-aminobenzylpenicillins, such as ampicillin, are known for oral administration only incompletely absorbed.

The object on which the invention is based

consisted in getting «-aminobenzylpenicillins, the higher concentrations of the antibiotic Substance in body fluids and tissues make possible as 6- (D - «- aminophenylacetamido) -penicillanic acid (Ampicillin) and also enable the fastest possible release of the antibiotic substance.

The oc-aminobenzylpenicillin esters according to the invention have the general formula

A CH ₃

-CH - CO - NH - CH - CH

NH ₂

CO-N-

CH ₃

or are pharmaceutically acceptable acid addition salts thereof, in which R is a hydrogen atom or a fluorine atom, and R ^{1 is} one of the groups

CH ₃

Il

- CH- O - CR ⁴
R ⁵ O

I Il

- CH - O - C - O - R ⁴

wherein R ^{4 is} an alkyl group having 1 to 4 carbon atoms and R ^{5 is} a hydrogen atom, a methyl group or an ethyl group.

Acid addition salts of the -Aminobenzylpenicllinester can with inorganic acids, such as with hydrochloric acid, hydrobromic acid, hydriodic acid or sulfuric acid, or with organic acids, such as citric acid, tartaric acid or maleic acid. With regard to that Asymmetric carbon atoms in the side chain, the compounds according to the invention exist in two epimeric forms.

The Λ-aminobenzylpenicillin esters according to the invention are well absorbed orally and lead to higher blood concentrations of the corresponding aminobenzylpenicillin than if the latter as such correspond to -CH-COOR ¹

the amount used. The esters are only slightly hydrolyzed in water, but much more when they are are subjected to the influence of hydrolytic enzymes, such as those found in blood serum and in other human body fluids are present. This property of the esters is very important as they leads to a rapid release of the antibacterially active aminobenzylpenicillins from the esters, if this is absorbed from the alimentary canal or in some other way into the bloodstream or tissue fluids to be introduced.

The -Aminobenzylpenicillinester according to the invention are preferably administered orally, either as such or in the form of their salts, and they can also with solid support materials and / or auxiliaries be administered mixed.

The compounds of the invention are used in the treatment of bacterial infections in humans for example in amounts corresponding to 5 to 200 mg / kg / day, preferably in amounts of 10 to 100 mg / kg / day in single doses, for example 2 times, 3 times or 4 times per day, administered.

The manufacture of the oc-aminobenzylpenicillin ester according to the invention either takes place in a manner known per se

a) by reaction of an (X-substituted penicillin the general formula

<C- \ - V \\ - CONHCH- eil

CH,

CH,

or a salt thereof, in which A is an amino group or one which can be converted into an amino group Group, for example an azido group or

CO N CH-COOR "

Represents a nitro group or a halogen atom such as a chlorine atom or a bromine atom, and R ″ represents Means hydrogen atom or a cation with

a compound of the general formula XR ¹ , where X is a halogen atom or a group functionally equivalent thereto, such as an alkylsulfonyloxy or an arylsultonyloxy group and R has the above meaning or
b) by reacting a compound of the general formula

> L ^V

CH-CO-Y ¹
A.

nil an ester of 6-aminopenicillanic acid of alllemcincn formula

CH ₃

NH-CH-CH

O = C-

-N-

CH ₃
CH-COOR ¹

wherein R, A and R ^{1 have} the above meaning and the radicals -CO-Y ¹ and X'-NH- are those radicals which can react with one another to form the group - CO —NH-.
For example, Y ¹ can be hydrogen or an alkali metal atom or a tertiary ammonium group and X ^{1 can be} a halogen atom, preferably a chlorine atom or bromine, or a group functionally equivalent thereto, such as an alkylsulfonyloxy or arylsulfonyloxy radical.

The substituent A can be a group which can be converted to an amino group by methods which are sufficiently mild to avoid destruction of the molecule on the ester group or on the lactam ring. In particular, all those groups A which are commonly used in the synthesis of aminopenicillins can be used. The substituent A preferably has the general formula Z — NH—, in which Z is a benzyloxycarbonyl group, a para-halogen, a para-nitro or para-methoxybenzyloxycarbonyl group, a 0, /?, J3-trichloroethyloxy- ₄ s carbonyl group or may be a sulfur-containing radical such as a triphenymethylsulfenyl group or an arylsulfonyl group such as an ortho-nitrophenylsulfenyl group. Z can also be the triphenylmethyl radical or a group obtained by reacting the free amino group with a α-dicarbonyl compound, such as acetylacetone, an acetoacetic acid ester or benzoylacetone, with the formation of enamines or Schiff's bases. In general, it can be said that any Z group which can be removed by reduction, hydrolysis with weak acids or by other mild reactions known per se is suitable. The starting materials for the compounds in which A is different from NH ₂ are known as intermediates in the synthesis of Λ-aminobenzylpenicillin. They exist in two epimeric forms. If the starting materials are prepared in the form of the D- or L-epimer, the corresponding epimeric form of the compounds of the invention is obtained. On the other hand, if a mixture of the epimeric forms of the starting materials is used, a mixture will also be obtained in the end products.

The above reaction 3) can be carried out at or below room temperature or by careful heating to the boiling point of the solvent used, depending on the meaning of R ¹¹ and X. It is possible to use various organic solvents or mixtures thereof with water, such as Acetone, dioxane, tetrahydrofuran, methyl chloride or dimethylformamide. The reaction products are crystalline or oily products that can be used in the next step without further purification. Repeated reprecipitation can convert the oily products into crystalline or amorphous powders.

The optionally subsequent conversion of the group A into an amino group can, depending on the Meaning of A to be carried out according to different methods, from the peptide synthesis and Aminobenzylpenicillin synthesis are known to the person skilled in the art. Catalytic hydrogenation is preferred when A has the formula Z — NH - and Z has the benzyloxycarbonyl radical or a closely related radical or the triphenylmethyl radical. The hydrogenation takes place preferably at room temperature and either at atmospheric pressure or at a slightly elevated pressure Pressure in a solvent that is a non-reducing organic solvent or a mixture of one such solvent can be with water. The preferred catalysts are noble metal catalysts, such as palladium or platinum catalysts or Raney nickel, but others can also Catalysts are used. Electrolytic reduction can also be used in these cases. When Z is a ß.jS.jS-trichloroethyloxycarbonyl group means, reduction with zinc in acetic acid is preferred. A hydrolysis with a weak acid is preferred when Z is a sulfur-containing radical, the radical of an enamine or a Schiff base means such as hydrolysis at a pH of about 2 in a dilute solution of Hydrochloric acid in aqueous acetone. As is known from the literature, an o-nitrophenylsulfenyl radical can also be removed by a nucleophilic attack on the sulfur atom of the sulfenamide group, and in this case the best results are obtained with potassium iodide or sodium iodide, sodium thiosulfate, Sodium hydrogen sulfide, sodium hydrogen sulfite or potassium thiocyanate. Instead, the sulfenamide compound also reacted with a thiophenol in an organic solvent such as dimethylformamide will. Other sulfenamide residues are removed in the same way. When A is an azido or nitro group is, such groups can be converted into the free amino group in a manner known per se by the azido and nitro groups are catalytically hydrogenated with a noble metal catalyst or with Raney nickel or is subjected to electrolytic reduction. When A is a halogen atom such as a Is bromine, it is by amination, such as with hexamethylenetetramine, into a Amino group converted.

In reaction b), for example —CO — Y ^1, the radical of an acid halide, such as an acid chloride or acid bromide, an anhydride or a mixed anhydride with an alkylcarboxylic acid, such as isobutylcarboxylic acid, a carboxylic acid, an inorganic acid or a sulfonic acid or else a The remainder that can be obtained by reacting the α-substituted phenylacetic acid and

of a carbodiimide or Ν, Ν'-carbonyldiimidazole. X ¹ can be a hydrogen atom or a trialkylsilyl group in which the alkyl group has no more than 5 carbon atoms. The reaction can be carried out in an organic solvent or in a mixture of such a solvent with water either at a low temperature or at a slightly elevated temperature. Suitable solvents are methylene chloride, chloroform, ethyl acetate, acetone, dimethylformamide or diethylacetamide, ether, tetrahydrofuran or dioxane. The reaction products are isolated in a conventional manner, such as by repeated reprecipitation or by removal of the solvent and subsequent recrystallization from a solvent.
The compounds of the general formula

S CH,

X ¹ -NH-CH-CH C

cn.,

O- C N- -CH-- COO -R ¹

are new connections. They can be obtained by reacting 6-aminopenicillic acid in the form of a salt, such as an alkali metal salt or a triethylammonium salt, with a halogen alkyl ester of the general formula XR ¹ , in which X is a halogen atom, preferably a chlorine atom or bromine atom, or a functionally equivalent group, like a sulfonyloxy radical, means. The 6-aminopenicillic acid can be used as such, or the 6-amino group in it can also be protected by a protective group during the esterification. In this case, however, only those protective groups are suitable which can easily be removed without weakening the lactam ring or the ester group, such as triphenylmethyl radicals or trimethylsilyl radicals. The reaction takes place in an inert organic solvent, such as for example. wise acetone, dimethylformamide or methylene chloride and at or below room temperature or at a slightly elevated temperature. When the amino group is protected by a protecting group, the protecting group can be removed in various ways, such as by hydrogenolysis or hydrolysis under neutral or acidic conditions in which the lactam ring and the ester group are not attacked. The reaction products are expediently isolated in the form of their acid addition salts, for example with paratoluenesulfonic acid or other inorganic or organic acids such as sulfuric acid, phosphoric acid, hydrochloric acid, acetic acid, maleic acid or tartaric acid.
The compounds of the general formula

S CH,

/ \ /
X ¹ NH-CH-Cl! C.

cn,

OC --N-- CH-CUO R ¹
can also be prepared by esterification of commercially available penicillins or, preferably, their salts with a compound of the above formula XR ¹ under similar conditions as described, after which the side chain of the penicillin ester obtained ^ is split off. Benzyl penicillin or phenoxymethyl penicillin is preferably used. The amide group can be split off with the aid of a modification of the process described in Belgian patent 6 98 596, in which 6-acylaminopenicillanic acid ester is reacted with an acid halide in the presence of an acid-binding agent such as quinoline or pyridine. However, the preferred acid halide is phosphorus pentachloride since the reaction in this case is at a low temperature

ι «, what the stability of the increased intermediate formed, which is likely an iminohalide. The response can be in various solvents, but the preferred solvents are chloroform

ίο and methylene chloride.

The intermediate is not isolated, but with an excess of primary alcohol to form treated with an iminoether. The reaction temperature and reaction time depend on the one used

is alcohol off. In most cases, temperatures between -20 and 20 ^° C. are suitable.

The imino ether is also not isolated, but subjected to acidic alcoholysis and hydrolysis, which causes a cleavage of the C-N bond, wherein the

ίο corresponding 6-aminopenicillanic acid esters formed will. Instead, the side chain can also be cleaved enzymatically in a manner known per se, such as is described, for example, in French patent specification 15 76 027. In the case of the generally used

.15 th methods, the esters of 6-aminopenicillanic acid from the reaction mixture as such or in the In the form of salts with inorganic or organic acids, for example in the form of the hydrochloride or tosylates.

Preferred compounds according to the invention are

the compounds of Examples 1, 6, 9, 13, 14, 15, 16 and

Ethoxycarbonyloxymethyl-6- (D- «- amino-m-fluoro-4S phenylacetamidoj-penicillanate as well

Ethoxycarbonyloxymethyl 6- (D - «- amino-p-fluorophenylacetamido) penicillanate.

example 1

so ^v

l'-acetoxyethyl 6- (D-a-aminophenylacetamido) penicillanate

<; _ <; a) The sodium salt of 6- (D-oc-Azidophcny! acetamido) -penicillanic acid (96%, 12.4 g, 0.03 mol) was dispersed in dry dimethylformamide (30 ml) and while stirring and cooling with ice with -chlorophyl acetate (3.7 g, 0.03 mol) dissolved in dry dimethylformamide

( _{1 (J} (15 ml), added. After 30 minutes the stirrer was continued without cooling overnight. The reaction mixture was poured into saturated sodium bicarbonate solution (500 ml) to give a white emulsion. This was washed with water ( 400 ml) diluted unc

(, <; extracted three times with ether. The combined etherex tracts were washed with water and saturated sodium chloride solution and dried. Evaporation de Ether solution in vacuo gave the a-acetoxyethyl ester

709 650/12

of e-iD-Ä-azidophenylacetamido-penicillanic acid (3.1 g) with a purity of 58% (hydroxylamino sample). It had a strong IR absorption at 2100cm- ' and at 1760 cm- 'indicating the presence of an azido group and an O-lactam ring. s

b) α, - Acetoxyethyl-6- (D - «- azidophenylacetamido) penicillanate (58%, 2.8 g), dissolved in ethyl acetate (40 ml), was mixed with 0.2 M NaH ₂ PO ₄ (40 ml ) treated, acidified to pH 2.2 with 2-M hydrochloric acid and hydrogenated over a palladium-on-charcoal catalyst under ambient conditions for 2 hours. The catalyst was removed by filtration and washed with ethyl acetate and buffer. The organic phase of the combined filtrates was separated off and extracted with water at pH 2.1. The combined aqueous extracts were washed with ether, covered with a layer of ethyl acetate and neutralized by adding 2N sodium hydroxide with stirring. The aqueous phase was separated off and extracted once with ethyl acetate. The combined ethyl acetate extracts of the neutralized aqueous solution were washed with brine and water and extracted twice with water at pH 2.5 by adding 2M hydrochloric acid. The combined aqueous extracts were washed with ether and concentrated in vacuo at 25 ° C., an oily residue being obtained which was dissolved in isopropanol and repeatedly evaporated in vacuo and the hydrochloride of l'-acetoxy-

ethyl 6- (D - «- aminophenylacetamido) penicillanate
(1.1 g) slightly alj colored powder having a purity _of! Gives ₀ 100% (Hydroxylaminprobe). It inhibited the growth of staph. aureus Oxford at a concentration of 0.32 μg / ml, of E. coli and P. mirabilis at 50 μg / ml.

c) l'-Acetoxyethyl-6- (D - «- azidophenylacetamido) penicillanate [strong IR absorption at 2100 cm- '(azido group) and at 1780 to 1760 cm-' (/? - lactam, ester)] has been eliminated Sodium 6- (Da-azidophenylacetamido) penicillanate (397 g, 1 mole), & chloroethylacelate (368 g, 3 moles), and sodium bicarbonate (504 g, 6 moles) _4C . The azido ester in ethyl acetate (800 ml) was hydrogenated at ambient conditions over a catalyst (25 g) of palladium (5%) on charcoal. The catalyst was removed by filtration and washed with ethyl acetate. The combined filtrates were extracted with water at pH 2.5 by adding dilute hydrochloric acid. Lyophilization of the aqueous phase gave the hydrochloride of 1'-acetoxyethyl 6- (D - «- aminophenylacetamido) penicillanate (110 g), mp 160 to 164 ° C (decomposition), [«] · ,; + 184 ° C (C = I ₁ CHCl ₃ ); so + 179.9 (C = 1, 50% isopropanol). It showed strong IR absorption at 1780 to 1755 cm- ', indicating the presence of j3-lactam and ester carbonyl, and inhibited the growth of Staph. aureus Oxford at a concentration of 0.25 μg / ml. *, <,

d) A suspension of dicyclohexylammonium-a-ionitrophenylsulfenyl) aminophenyl acetate (4.9 g, 0.01 mol) and the hydrochloride of r-acetoxyethyl-6-aminopenicillanate (3.4 g, 0.01 mol) in chloroform (60 ml) was stirred overnight. Dicyclohexylcarbodiimide (2.2 g, 0.01 <, <> mole) was added and stirring was continued for 6 hours. The reaction mixture was filtered and washed with water, dilute sulfuric acid, 1N potassium bicarbonate and water, dried and evaporated to dryness in vacuo. The yellow back, _'s stand was treated with ethyl acetate, filtered and evaporated again. The residue was recrystallized from ethyl acetate-petroleum ether and gave l'-acetoxyethyl 6- [D - «- (o-nitrophenylsulfenyl) aminophenylacetamido] penicillanate (1.5 g, mp = 122 bl · 125 ° C. This possessed a strong IR absorption at 178 (to 1750 cm- ', indicating the presence of ^ -lactam and ester carbonyl.

This penicillanate (1.2 g, 0.002 mol) was dissolved in 75% dioxane (17 ml) and brought to pH 3 Addition of 2 η-hydrochloric acid stopped. Sodium iodic (1.2 g, 0.008 mol) was added and the Gemiscr was stirred and adjusted to pH; held. After 20 minutes the iodine formed was mi 2 N sodium thiosulphate reduced and the mixture neutralized and treated with ethyl acetate after addition before Water extracted. The combined organic extracts were washed with water and added by adding of 2η hydrochloric acid to the stirred mixture up to it Extraction in water reached pH 2. The aqueous phase was washed with ethyl acetate and lyophilized and gave the hydrochloride before l'-acetoxyethyl-e ^ D-Ä-aminophenylacetatamidoJ-penicillanate (0.15 g). The product was made with the help of de; IR spectrum as that in Example Ib) produced « identified.

Example 2

By repeating the procedure of Example Ib, the following ester of 6- (D - «- aminophe was obtained nylacetamidoj-penicillanic acid hydrochloride from the ent speaking azido compounds.

'■■ slcr (ichall smallest Hcmmkon / cnlralion

(ug / mm)

Slaph. H. coli P. miraaurcus bilis

Oxford

Mcthoxycarhnnyl- 69 0.16 25 6.25

oxymelhyl ester

(Determination of the concentration according to the Hydroxamatme method, antibacterial activity, determined according to the Tube dilution method in broth)

All esters of Examples 1 and 2 were hydrolyzed with the formation of .delta.-iD-a-AminophenylacetamidoJ-penicillansäun by incubation (37 ⁰ C) with human Serurr.

Example 3

r-Propionyloxyethyl-6- (D-ix-aminophenylacctamido) penicillanate

Nalium 6- (D-α-azidophenylacetamido) penicillanate (6.0 g, 0.015 mol) (X-chloroethyl propionate (5.7 g, 0.04; Mol) and sodium carbonate (7.1 g, 0.084 mol) were at room temperature for 65 hours in 70% igcm dioxar (100 ml) stirred. The solid phase of the reaction mixture was filtered off and washed with dioxane. the combined filtrates were concentrated in vacuo and with a mixture of benzene (50 ml) unc saturated sodium bicarbonate solution (25 ml) The organic phase was separated, washed with brine, dried and evaporated in vacuo and gave l'-propionyloxyethyl-6- (D - «- azidophenyl ·

acetamido) penicillanate (1.3 g) as a yellow oil. This showed strong IR absorption at 2110 and 1780 bis 1750cm- ', indicating the presence of an azido group, the /? -Lactam ring and ester groups.

The ester was dissolved in ethyl acetate (50 ml) for 30 s at ambient conditions above 1 min Catalyst (1.3 g) of palladium (5%) hydrogenated on carbon. The catalyst was filtered off and with Washed ethyl acetate and water. The pH of the combined filtrates was determined by adding diluent ι ο ter hydrochloric acid adjusted to 2.2. The aqueous phase was separated off and the organic phase once with Extracted water at pH 2.2. The combined aqueous extracts were lyophilized to give the hydrochloride of l'-propionyloxyethyl-6- (D- «- aminophenylacetamidoj-penicillanate (0.6 g) as an amorphous solid.

The product showed strong j3-lactam and ester carbonyl bands at 1760 cm- ¹ in its IR spectrum. It inhibited the growth of staph. aureus Oxford ia at a concentration of 0.63 ug / ml.

Example 4

l'-Isobutyroxyethyl and l'-Vaieryloxyethyl-6- (D-a-aminophenylacetamido) penicillanate

Repetition of the procedure according to Example la), but replacing "-Chloräthylacetat by ct -chloräthylisobutyrat and" -Chloräthylvalerat gave _ ₁₀ the intermediate product Γ-Isobutyroxyäthyl- and 1'-VaIeryloxyäthyl-e ^ da-azidophenylacetamidoJ-peniciiianat.

The compounds were characterized by their IR spectra, the strong absorption bands at 2100 cm- 'and 1780 to 1730cm-', which of the (s Azido group, the ß-lactam ring and the ester groups are to be ascribed. In the hydrogenation according to the example Ib) they gave l'-Isobutyroxyäthyl- and l'-Valeryloxyäthyl-6- (D - «- aminophenylacetamido) -penicilIanat, which were isolated as hydrochlorides. The amino esters had strong α-lactam and ester absorption in theirs IR spectra around 1760 cm - '.

45

Example 5

Methoxycarbonyloxymethyl 6- (D-α-aminophenylacetamido) penicillanate

a) By replacing the a-chloroethyl acetate with monochlorodimethyl carbonate (3.7 g, 0.03 mol) in example la), but otherwise under the same working conditions as in this example, the methoxycarbonyloxymethyl ester of 6- (da-azidophenylacetamido) - was obtained penicillanic acid (6.6 g) with a purity of 76.5% ^ s (hydroxylamine sample). The product showed IR absorption at 2100 cm- '(Nj group) and 1765 cm ¹ (β-lactam ring).

b) Meihoxycarbonyloxymethyl-e-iD-iX-aminophenylacet- r «, amido) -penicillanate was prepared according to the method of Example 2 by converting sodium e-iD-Ä-azidophenylacctamidoJ-penicillanate (24.2 g, 0, 06 mol) and chloromethyl methyl carbonate (6.2 g, 0.06 mol) ran out. The compound was isolated as the hydrochloride (5.6 g), m.p. = 145 to 150 ⁰ C (decomposition), ₍ , _s [tx] V = + 182.5 (CI, CHCIj), +191.2 (C = , 50% isopropanol). It showed strong IR absorption at 1780 to 1760 cm - ', which was due to the presence of j3-lactam. The product inhibited the growth of Staph. Aureus Oxford at a concentration of 0, 13 µg / ml.

Example 6

l'-Äthoxycarbonyloxyäthyl-6- (D - «- aminophenylacetamido) -penicillanate

a) The sodium salt of 6- (D - «- Azidophenylacetamido) penicillanic acid (4.0 g, 0.01 mol) was dispersed in dry dimethylformamide (10 ml) and with stirring at 50 ⁰ C with« -chlorodiethyl carbonate (1.5 g , 0.01 mol) dissolved in dry dimethylformamide (6 ml). After stirring at 50 ° C. for 20 hours, the reaction mixture was cooled and poured into an ice-cold saturated sodium bicarbonate solution and extracted three times with ether. The combined ether phases were washed with water and brine and dried. Evaporation in vacuo gave the ethoxycarbonyloxyethyl ester of 6- (D - - azidophenylacetatamido) penicillanic acid (2.6 g) in the form of a yellow-brown glass. It had strong IR absorption at 2100 cm- 'and at 1750 cm-', indicating the presence of an azido group and a jU-lactam ring and an ester group.

b) The Λ-ethocycarbonyloxyethyl ester of 6- (D-oc-azidophenylacetamido) penicillanic acid (2.5 g) was dissolved in ethyl acetate (40 ml), and 0.4 mol of NaH ₂ PO ₄ , acidified with 2 M hydrochloric acid ( 40 ml) to pH 2.2, and 4 g of a catalyst of 10% palladium on carbon were added, and then hydrogenation was carried out at normal pressure for 0.5 hour. The catalyst was filtered off and washed with ethyl acetate and buffer solution. The filtrate was separated and the organic phase was extracted with water at pH 2.1. The combined acidic water phases were washed with ether, ethyl acetate was added and the mixture was neutralized with 2M NaOH while stirring. The phases were separated and the water solution was extracted again with ethyl acetate. The two ethyl acetate extracts were combined, washed with brine and water and extracted with water at pH 2.5 by adding 2M hydrochloric acid. The extraction of the ethyl acetate phase was repeated once. The combined water extracts were washed with ether and concentrated in vacuo at 25 ° C. The semi-solid residue was dissolved in isopropanol and concentrated again, repeating the procedure once. The residue was stirred with ether, filtered and washed with ether and gave-ethoxycarbonyloxyethyl ester of 6- (Da-aminophenylacetamido) penicillanic acid hydrochloride (0.8 g) as a yellow-white powder with a purity of 86.4% (hydroxylamine sample).

The product inhibited the growth of staph. aureus Oxford at a concentration of 0.13μg / ml and E. coli at a concentration of 125μg / ml (on agar) and contained in its IR spectrum a strong absorption band at 1750 cm- ¹ , indicating the presence of a j3-Lactamririges and an ester group.

Example 7

r-Äthoxycarbonyloxyäthyl-e-iD-oc-aminophenylacetamido) penicillanate

r-Äthoxycarbonyloxyäthyl-G-iD-a-u / .idophenylacetamido) -penidllanal (98 g) was made from Nalrium-6- (D- <x-

azidophenylacetamido) penicillanate (397 g, 1 mol), Λ-chlorodiethyl carbonate (458 g), 3 mol) and sodium bicarbonate (504 g, 6 moles). It showed strong IR absorption at 2090 cm- 'and at 1780 bis 1750 cm- ', indicating the presence of an azido group, a 9-lactam ring and ester carbonyl groups indicated.

The compound was dissolved in ethyl acetate (700 ml) and at ambient conditions over 18 g of catalyst hydrogenated with 5% palladium on charcoal. The catalyst ro was filtered off and washed with ethyl acetate. The combined filtrates were washed with water at pH 2.5 Extracted by addition of dilute hydrochloric acid. Lyophilization of the aqueous phase gave the hydrochloride of

1 '-Athoxycarbonyloxyäthyl-ö ^ Da-aminophenylacet- ι s amido) penicillanate (94 g). F. = 171 to 176 ° C (decomposition), [«]» = +161.5 (C = 1, CHCl ₃ ), +171.8 (C = 1, 50% isopropanol). The product showed strong IR absorption at 1780 to 1750 cm- 'due to the j3-lactam ring and the ester carbonyl groups. It ^ o inhibits the growth of staph. aureus Oxford at a concentration of 0.13 μg / ml.

Example 8

l'-Äthoxycarbonyloxyäthyl-6- (D - «- aminophenylacetamido) -penicillanate

a) To a suspension of the hydrochloride of a-aminophenylacetyl chloride (2.6 g, 0.0125 mol), stirred yo in dry nitrogen, were sodium carbonate (1.1 g, 0.0125 mol) and hydrochloride of l'-ethoxycarbonyloxyethyl 6-aminopenicillanate (3.3 g, 0.01 mol) added. After stirring for 90 minutes, the mixture was filtered and isopropanol (15 ml) was added to the filtrate. Evaporation in vacuo at room temperature gave an oily residue which, on dilution with petroleum ether, resulted in precipitation of the hydrochloride of l'-ethoxycarbonyloxyethyl 6- (D - «- aminophenylacetamido) penicillanate (1.5 g). The product was purified by precipitation from acetone-petroleum ether.

b) Sodium N- (1-methoxycarbonylpropen-2-yl) - «- aminophenyl acetate (2.5 g, 0.0092 mol) and N-methylmorpholine (0.05 ml) in dry acetylacetate (40 ml) was stirred and washed at -15 ° C with isobutyl chlorofomate (1.4g, 0.01 mol) treated. After 6 minutes it was

1 '-ethoxycarbonyloxyethyl-e-aminopenicillanate (0.01 Mol) in ethyl acetate (20 ml) was added dropwise while stirring continued and the temperature increased - 15 ° C was kept. Ten minutes after the addition was complete, the cooling bath was removed and stirring was started continued for another 45 minutes. The reaction mixture was washed with water, 0.5-M sodium bicarbonate solution and washed again with water, dried and concentrated in vacuo to give an oily residue, which was treated with tetrahydrofuran (20 ml) and water (20 ml). The mixture was through Addition of 2η hydrochloric acid acidified to pH 2.5. In 30 minutes, most of the tetrahydrofuran was do in vacuo removed and the acidic aqueous phase washed with ethyl acetate and evaporated to give the hydrochloride of l'-Äthoxycarbonyloxyäthyl-e-iD-ix-aminophcnylacetamido) penicillanate (0.35 g) as an amorphous solid which can be identified as the product obtained above with the aid of (> s its IR spectrum has been identified.

The solution used of I'-Äthoxycarbonyloxyäthyl-6-aminopenicilInnat was obtained by neutralizing an aqueous solution of its hydrochloride and extracting it with ethyl acetate.

Example 9

Ethoxycarbonyloxymethyl 6- (D - «- aminophenylacetamido) penicillanate

A stirred suspension of sodium 6- (D - «- azidophenylacetamido) penicillanate (12.1 g, 0.03 mol) in dry dimethylformamide (30 ml) was treated with chloromethyl ethyl carbonate (4.2 g, 0.03 mol) in dry dimethylformamide (15 ml) in an ice bath. After 45 minutes the cooling bath was removed and stirring continued overnight. The dark one The reaction mixture was poured into twice its volume of a saturated sodium bicarbonate solution, and the solid phase formed was filtered off and washed with water and ether. The United Filtrates were extracted with ether. The ether extracts were washed with water, dried and im Evaporated in vacuo at room temperature and gave

Ethoxycarbonyloxymethyl-e-iD-a-aminophenylacetamido) penicillanate (4.7 g) as a brownish oil. The product showed strong absorption bands in the IR spectrum at 2120 cm- 'and at 1780 to 1750 cm-', referring to the azido group and the jS-lactam ring and ester carbonyl groups were due.

The azidoester (2.6 g) was dissolved in 60% ethanol (50 ml) and dried at 4 atm (60 psi) for 30 minutes hydrogenated a prehydrogenated Raney nickel catalyst. The catalyst was filtered off and washed with ethanol washed. The filtrate was diluted with water and extracted with ethyl acetate. The united organic Extracts were washed with water and then with water with the addition of dilute hydrochloric acid until Extracted reaching a pH value of 2.5. The aqueous phase was separated off and lyophilized and gave the crystalline hydrochloride of l'-ethoxycarbo-

nyloxymethyl-e-p-a-aminophenylacetamidoj-penicillanate (0.7 g). It showed strong IR absorption at 1760 cm- 'and inhibited the wax staph. aureus Oxford at 0.05 μg / ml.

Example 10

Propoxycarbonyloxymethyl-6- (D - «- aminophenyl · acetamidoj-penicillanate

Following the procedure of Example 9, propoxycarbonyloxymethyl-e-iD-a-azidophenylacetamido) penicillanate (5.2 g) from sodium 6- (D- <x -azidophenylacetamido) penicillanate (12.1 g, 0.03 mol) and chloromethyl n-propyl carbonate (4.6 g, 0.03 mol). It showed strong IR absorption at 2120cm- 'and at 1780-1740cm-', suggesting the presence of a Azido group and from j3-lactam and ester carbonyl groups was due.

^ When hydrogenated over Raney nickel, this ester gave (2.9 g) propoxycarbonyloxymethyl-e-iD-a-aminophenylacetamido) penicillanate, which was isolated as the hydrochloride (1.0 g). It showed strong ^ -lactam- and Ester absorption bands in the IR spectrum at 1775 cm- ' and inhibited the growth of staph. aureus Oxford at a Knn7pmriitii-> n ,, "" nm - / _ i

Example 11

Isopropoxycarbonyloxymethyl-ö-iD-a-aminophenylacetamido) penicillanate

In the manner described in Example 17, isopropoxycarbonyloxymethyl-e-iD-a-azidophenylacetamido) penicillanate (7.2 g) was prepared from sodium 6- (D- «- azidophenylacetamido) penicillanate (9.9 g, 0.025 mol ) and chloromethyl _{isopropyl carbonate (3.8 g s} 0.025 mol). Its IR spectrum contained strong absorption bands at 2110 cm- '(azido group) and 1780 to 1740 cm-' (jJ-lactam and ester carbonyl).

Hydrogenation over Raney nickel converted the ester (4.4 g) into isopropoxycarbonyloxymethyl-e-fD-a-aminophenylacetamido) penicillanate which was isolated as the hydrochloride (1.3 g). It showed strong absorption in the infrared spectrum at 1760 cm- ', which is characteristic for the / J-lactam ring and ester carbonyl groups. The product inhibited the growth of staph. aureus Oxford at a concentration of 0.05 μg / ml.

Example 12

Butoxycarbonyloxymethyl 6- (D - «- aminophenylacetamido) penicillanate

Following the procedure of Example 9, butoxycarbonyloxymethyl 6- (D - «- azidophenylamido) penicillanate was made (6.0 g) from sodium 6- (D - «- azidophenylacetamido) penicillanate (9.9 g, 0.025 mol) and chloromethyl-n-butyl carbonate (4.2 g, 0.025 mol). the Compound showed strong IR absorption at 2120 cm- ' (Azido group) and at 1780 to 1750 cm- '(0-lactam and ester carbonyl).

It (2.9 g) was hydrogenated over Raney nickel and gave butoxycarbonyloxymethyl-e ^ D-a-aminophenylacetamido) penicillanate, which was isolated as the hydrochloride (1.0 g). It showed strong IR absorption at 1780 bis 1770 cm- '(j3-lactam and ester carbonyl) and inhibited the growth of Staph. aureus Oxford at one Concentration of 0.05 μg / ml.

Example 13

r-Acetoxyethyl 6- (D - «- amino-m-fluorophenylacetamido) penicillanate

The hydrochloride of r-acetoxyethyl-6-aminopenicillanate (3.4 g, 0.01 mol) in dry dimethylformamide (50 ml) was stirred in an ice bath and treated with N, N-dimethyl aniline (2.4 g, 0.1 mol). 02 mol) and then treated with a solution of oc- azido-m-fluorophenylacetyl chloride (2.1 g, 0.01 mol) in dry ether (5 ml). After stirring for 1 hour, water (100 ml) was added and the mixture was adjusted to pH 6.5 and extracted three times with ethyl acetate. The combined organic extracts were washed with water and brine, dried and evaporated in vacuo and gave r-acetoxyethyl-6-

(D - «- azido-m-fluorophenyiacetamido) penicillanate
(1.5 g) as an oily residue. This product showed strong IR absorption at 2110 and 1780 to 1750 cm 'indicating the presence of the azido group and of ^ -lactam and ester carbonyl groups.

This compound (0.7 g) was dissolved in 60% ethanol (7 ml) and added to Rancy nickel catalyst (0.7 g) in 80% ethanol (5 ml) was added and the mixture was hydrogenated at ambient temperature for 30 minutes. The catalyst was filtered off and washed with ethanol. The combined filtrates were added in vacuo at 35 ° C concentrated to an oily residue which was dissolved in ethyl acetate. Water (20 ml) was added and the pH was adjusted to 1.6 by adding 2η hydrochloric acid to the stirred mixture. The aqueous phase was separated off and evaporated in vacuo at 35 ° C. and gave the hydrochloride of l'-acetoxyethyl-e-iD-a-amino-m-fluorophenylacetamidoJ-penicillanate (0.4 g) as a crystalline residue.

The product inhibited the growth of staph. aureus Oxford at a concentration of 0.25 μg / ml and contained a strong β-lactam absorption band at 1760 cm- 'in its IR spectrum.

Example 14

l'-Ethyloxycarbonyloxyethyl-6- (D - «- aminom-fluorophenyl? .cetamido) -penicillanate

To sodium 6- (D-a-azido-m-fluorophenylacetamido) penicillanate (4.2 g, 0.01 mol) and sodium bicarbonate (5.1 g, 0.06 mol) in 70% dioxane (15 ml) «-Chlorodiethyl carbonate (4.6 g, 0.03 mol) was added, and the mixture was 2 days at room temperature touched. The reaction mixture was filtered, the residue washed with dioxane and the combined Filtrates were evaporated to dryness in vacuo. The residue was in a mixture of benzene (40 ml) and saturated aqueous sodium bicarbonate solution (20 ml). The organic phase was separated, washed with brine, dried and evaporated and gave 1'-Äthyloxycarbonyloxyäthyl-6-

(Da-azido-m-fluorophenylacetamidoJ-penicillanate
(1.0 g). The compound showed strong IR absorption at 2100 and 1750 cm- ', which was due to the azido group and ^ -lactam and ester groups.

It was dissolved in ethyl acetate (10 ml) and made into a Hydrogenation catalyst (0.5 g) composed of 5% palladium on charcoal, which had been pre-hydrogenated in ethyl acetate (5 ml), added and hydrogenated for 2 hours under ambient conditions. The catalyst was filtered off and with Washed ethyl acetate. Water (10 ml) was added to the combined filtrates and the pH was lowered adjusted to 2.2 by adding 2η hydrochloric acid to the stirred mixture. The aqueous phase was

separated, washed with ether and lyophilized and gave the hydrochloride of l'-Äthyloxycarbonyloxyäthyl-e-iD-a-amino-m-fluorophenylacetamidoJ-penicillanate (0.2 g) as a white crystalline residue.

The product contained a strong absorption band at 1780 cm- 'in its IR spectrum, correspondingly a /? - lactam ring. It inhibited the growth of Staph. aureus Oxford at a concentration of

Example 15

l'-Acetoxyethyl 6- (D - «- amino-p-fluorophenylacetamido) penicillanate

Using the procedure of Example 14, 1'-acetoxyethyl 6- (D-a-azido-p-fluorophenylacetamido) penicillanate was obtained (2.2 g) from sodium 6- (D - «- azi-

do-p-fluorophenyiacetamidoj-penicillanate (4.2 g, 0.01 mol), α-chloroethyl acetate (3.7 g, 0.03 mol) and sodium bicarbonate (5 g, 0.06 mol). The IR spectrum of the product showed strong absorption bands at 2110 cm- '(azido group) and at 1775 to 1750 cm ^-1 (/? - lactam and ester carbonyl).

The ester was hydrogenated over Raney nickel catalyst and gave the hydrochloride of l'-acetoxyethyl

e-iD-Ä-amino-p-fluorophenylacetamidoJ-penicillanate (1.5 g) as light yellow crystals. The product had strong ^ -lactam and ester absorption bands in its IR spectrum at 1765cm- 'and inhibited the growth of Staph. aureus Oxford at one Concentration of 0.25 μg / ml.

Hydrogenated ambient conditions. The catalyst was filtered off and washed with ethyl acetate. Water (10 ml) was added to the combined filtrates and the pH was adjusted by adding 2N hydrochloric acid adjusted to 2.2 to the stirred mixture. The aqueous phase was separated off with Washed ether and evaporated to dryness in vacuo and gave the hydrochloride of l'-Äthyloxycarbonyloxyäthyl-e-fD-a-amino-p-fluorophenylacetamido) penicillanate (0.2 g) as white crystals. The product had a strong IR spectrum β-lactam absorption band at 1760 cm- 'and inhibited the growth of Staph. aureus Oxford at one Concentration of 0.25 μg / ml.

Example 16

1'-Ethyloxycarbonyloxyethyl-ö-fD-a-aminop-fluorophenylacetamido) penicillanate

To sodium 6- (D - «- azido-p-fIuorphenylacetamido) -penicillanat (4.2 g, 0.01 mol) and sodium bicarbonate (5.1 g, ₁ OS O mole) in 70% dioxane (15 ml ) «-Chlorodiethyl carbonate (4.6 g, 0.03 mol) was added, and the mixture was stirred at room temperature for 2 days. The reaction mixture was filtered, the residue washed with dioxane, and the combined filtrates were evaporated to dryness in vacuo. ^ ₀ The residue was dissolved in a mixture of benzene (40 ml) and saturated aqueous sodium bicarbonate solution (20 ml). The organic phase was separated, washed with brine, dried and evaporated in vacuo and gave Γ-Äthyloxycar-

bonyloxethyl 6- (D-a-azido-p-fluorophenylacetamido) penicillanate (1.9g).

The product showed strong IR absorption at 2110 and 1750 cm - ', corresponding to the azido group and the j9-lactam ring and the ester groups. It was dissolved in ethyl acetate (10 ml) and prehydrated Catalyst (0.5 g) of palladium (5%) on charcoal in ethyl acetate (5 ml) was added and taken for 2 hours Comparative examples

a) Oral absorption of the λ-aminobenzylpenicillin esters in humans

In clinical trials, the oral absorption of the following three compounds was tested in six healthy male patients.
I. 1 '-Athoxycarbonyloxyäthyl-e ^ Da-aminophenyl-

acetamido) penicillanate,
11. 1 '-Acetoxyäthyl-ö-iD-a-aminophenylacetamido) penicillanate,

III. Pivampicillin [pivaloyloxymethyl-6- (D-α-aminophenylacetamido) penicillanate].

Compounds I and II were in tablets, compound III in capsules in the form of a commercially available preparation. The amounts of the three esters corresponded to 250 mg of free ampicillin each. The tablets were given on an empty stomach. Blood samples were taken at predetermined time intervals removed and with regard to their content of ö-iD-a-aminophenylacetamidoJ-penicillanic acid below Analyzed using a microbiological test method. The following middle Serum level ^ g / ml) with standard deviation from Mean found:

Conclusion 0.5 1 6.05 1.5 2 1.96 1.04 4th 0.62 6th 0.22 X 0.063 7.65 ■ 1 0.63 3.10 to, 18 ■ to, 11 + 0.10 1 0.057 + 0.016 I 1.42 6.57 ■ t 0.28 2.65 1.13 0.68 0.20 0.063 I. 9.52 I 1.31 4.33 + 0.35 tO, 15 to, 11 l · 0.043 J 0.011 i 2.02 4.55 h0.50 2.70 1.26 0.73 0.20 0.068 11th 3.58 i 0.56 4.50 1 0.38 I 0.26 to, 16 I 0.050 I 0.018 I 0.80 1 0.70

b) Hydrolysis of the Λ-aminobenzylpenicillins to ( ₁₀₎ the corresponding aminobenzylpenicillins in buffer and in the presence of human serum

The hydrolysis of compounds according to the invention was carried out in buffer solution with and without human Serum determined. They were heated for 30 minutes at 37 ° C at, ·, ·, a concentration of 10μg / ml in Sörensen phosphate buffer (pH 7.4) and in another experiment in the same buffer with a content of 10% incubated with human serum. The test solutions were extracted with ethyl acetate to be unhydrolyzed To remove esters and were microbiologically analyzed for their aminobenzylpenicillin contents. Blank values were obtained by dissolving the esters in phosphate buffer at the same concentration and carrying out the extraction immediately. The values in the table below are determined by the Corrected blank values.

2! 44 457

S CH,

K-CH CO NH CH CH

-CH.,

CO - N CH-COOR ¹

Example no.

"Ai hydrolyzes after 30 minutes at 37 ° C

PhosphalpuCfer

I phosphate buffer 1! () "/" meal. serum

Pivampicillin CH ₃ 'H-OCOCH ₃ Ib 5.4 10.2 "CH-OCOCfJ, CH ₃ Q ₁ HY CH ₃ 3 7.0 17.0 -CH-OCOC ₂ H ₅ ΊI OCOOC ₂ H ₅ C ₆ H ₅ CH ₃ 4th 6.8 17.6 -CH-OCOCH (CH ₃ J ₂ C ₆ H ₅ CH ₃ 4th 5.2 12.1 -CH-OCOC ₄ H ₉ 2 Q ₁ H ₅ -CH ₂ OCOOCH ₃ 4.9 14.3 C ₆ H ₅ CH ₃ 6th 24.8 77.9 -CHOCOOC ₂ H ₅ 9 C ₆ H ₅ -CH ₂ OCOOC ₂ H ₅ 10 3.8 57.1 C ₆ H ₅ -CH ₂ OCOOC ₃ H ₇ 11th 13.3 64.7 C ₆ H ₅ -CH ₂ OCOOCH (CH ₃ J ₂ 12th 14.5 53.7 C ₆ H ₅ "CH ₂ OCOOC ₄ H ₉ 15.2 68.4 Q ₁ H ₅ CH ₃ 13th 13.0 41.1 "CH-OCOCH ₃ mF-C ₆ H ₄ cn., 14th 3.1 16.4 CH-OCOOC ₂ II ₅ 15th mF-C ₆ H ₄ C.
- ( 2.6 37.7 PFC ₆ H ₄ 16 9 14.3 PF C ₆ H ₄ 1.4 20.2

c) The acute toxicity of compounds after the physiological saline solution, the animals in a

Invention and injected by pivampicillin at intravenous (, $ lateral coccyx vein, and the mice were

Administration to mice was determined. Groups observed for 7 days after administration. the

Ten animals were used for each dose and deaths occurred within 2 hours of each

Compounds were given as hydrochlorides, dissolved in the administration.

21

R-CH-CO-NH-CH-CH C

NH ₂ · HCl

R '

CO-N

Pivampicillin

CH ₃

CH ₃

CH, CH —COOR '

LH ₅₁ , (mg'k iv, Mii use

80 100 *)

C ₆ H ₅ -QH ₅

C _n H,

-CH-OCOCH ₃ 200 400 **)

*) 50% Lclulilül at both concentrations.
**) The DL ₅₀ -WeI "! Lies between these limits.

LD ₅₁ , (mg / kg) iv. Miiusc

400 500 **) 760

-CH ₂ OCOOCH ₃
-CH ₂ OCOOC ₂ H ₅

CH ₃
-CH-OCOOC ₂ H ₅ 200 400 **)

-CH ₂ OCOOC ₂ H ₅ 700

*) The DLj "-Wcrl Vicgl between these limit values.

Claims (4)

Claims: I. A-aminobenzylpenicillin ester of the general formula S CH, ^ H-CO-NH-CH-CH C NH, CH, = CN CH-COO -R ¹ and their pharmaceutically acceptable acid addition salts, in which R is a hydrogen atom or fluorine atom, and R ^{1 is} one of the groups CH, - CH-Ο —C —R ⁴ or —CH-Ο —C — Ο —R ⁴ wherein R ^{4 is} an alkyl group having 1 to 4 carbon atoms and R ^{5 is} a hydrogen atom, a methyl group or an ethyl group. 2. A compound according to claim I of the formula S CH ₃ CH-CO-NH-CH-CH C CH ₃ CH ₃ NH ₂ O = CN CH-CO -CH-OCCH ₃ O and its pharmaceutically acceptable acid addition salts c 3. A compound according to claim 1 of the formula S CH ₃ CH-CO-NH-CH-CH C \
CH, CH ₃ NH, O = -CN CH —C — O —CH — OCOC ₂ H, Il II ο ο and its pharmaceutically acceptable acid addition salts c. 4. Process for the preparation of compounds according to Claims I to 3, characterized in that that in a known manner a compound of the general formula S CH, - C H - C O - N H - C H - C H C X: O C N CH - COO ■ - R " in which A denotes an amino group or a group which can be converted into a group, R "denotes a hydrogen atom or a cation and R i has the above meaning with a compound of the illustrative funnel X-R ^s , in which X is a halogen atom or a functionally equivalent thereto Means group and R ^{1 has} the above meaning, converts or a compound of the general formula with a compound of the general formula S CH, / \
X '- NH-CH-CH C = C N- CH,
CH-CO-O-R ¹