GB1578128A

GB1578128A - Amidinopenicillanoyloxyalkyl amoxycillinates

Info

Publication number: GB1578128A
Application number: GB12849/76A
Authority: GB
Original assignee: Leo Pharmaceutical Products Ltd AS
Current assignee: Leo Pharma AS
Priority date: 1976-03-30
Filing date: 1976-03-30
Publication date: 1980-11-05
Also published as: SE7703634L; FR2346355A1; NZ183549A; AU2370677A; FR2346355B1; DK134577A; AU503971B2; DE2713683A1; JPS52118492A

Description

(54) AMIDINOPENICILLANOYLOXYALKYL AMOXYCILLINATES (71) We, LEO PHARMACEUTICAL PRODUCTS LTD. A/S (LIVENS KEMISKE FABRIK PRODUKTIONSAKTIESELSKAB), a Company organized under the Laws of Denmark, of DK-2750 Ballerup, Denmark, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement: The present invention relates to hitherto unknown esters of formula I below, to salts of the esters of formula I with pharmaceutically acceptable acids, to methods for producing said new compounds, to pharmaceutical preparations containing said new compounds, and to methods of treating patients suffering from infectious diseases using said new compounds.

The compounds of the invention, which are valuable in human and veterinary practice, have the formula I

in which Rl represents hydrogen or methyl, and n is an integer from 5 to 8, i.e. the grouping

represents 1-piperidyl, hexahydro-1H-azepin-1-yl, hexahydro-1(2H)-azocin-1-yl, or octahydro- tH-azonin- 1-yl.

The invention comprises all possible isomeric forms of the compounds of formula I, in which the asterisk in the side chain moiety indicates an asymmetric carbon atom, and where the "+" in the ester moiety indicates an asymmetric carbon atom in the case where Rl is methyl, whereas the 6-aminopenicillanic acid moiety has the configuration indicated in formula I. The presence of asymmetric carbon atoms gives rise to several diastereomeric forms, and the invention covers all such forms as well as mixtures thereof.

As stated above, the invention also relates to salts of the esters of formula I with pharmaceutically acceptable non-toxic acids, e.g. hydrochloric and hydrobromic acid, phosphoric acid, sulphuric acid, nitric acid, p-toluene-sulphonic acid, methanesulphonic acid, formic acid, acetic acid, propionic acid, citric acid, tartaric acid and maleic acid.

However, salts with other, in themselves therapeutically active acids, such as penicillins, cephalosporins, and fusidic acid, are also within the scope of the invention.

The compounds according to the invention are valuable in the treatment of bacterial infections and will, after administration, be hydrolyzed by enzymes present in blood and many tissues to p-hydroxyampicillin and a 6-amidinopenicillanic acid. It is known that the simultaneous presence of these two types of antibiotics in the organism leads to a broad spectrum effect, and that furthermore a synergistic effect is obtained.

It is known that salts of p-hydroxy-ampicillin which otherwise would be suitable for parenteral administration are unstable, and until now this penicillin therefore has not been available for parenteral administration.

It has now surprisingly been shown that the compounds of formula I are stable and thus suitable for parenteral administration. The compounds are furthermore astonishingly non-toxic even when given parenterally. For instance the compound of Example 1 has an LD50 (i.p.) exceeding 1000 mg/kg.

The new compounds of formula I are thus intended for use in the parenteral treatment of patients suffering from infections where a quick response to the treatment is desirable. As they are absorbed from the gastro-intestinal tract, they may also be used enterally.

The parenteral administration of the compounds of the invention leads to immediate, high concentrations of p-hydroxyampicillin and the 6-amidinopenicillanic acid, a combination showing a pronounced synergy towards a number of pathogenic microorganisms, e.g.

Salmonella, Proteus, E. coli, Klebsiella, and Haemophilus influenzae, this selection not to be considered limiting to the present invention. A further advantage of such combination lies in the fact that bacterial organisms develop resistance less readily to such combination than to either of the compounds alone.

The invention also comprises a method for the preparation of the above described compounds. In a first embodiment, an ester of the formula II

in which formula Rl and n are as defined above, and X is a leaving group, e.g. chlorine, bromine, iodine, or p-toluenesulphonyloxy, is reacted with a salt of the formula III

in which R2 represents hydrogen or a protective group, such as benzyloxycarbonyl, triphenylmethyl, ss,ss,ss-trichloroethoxycarbonyl or 1-methyl-2-carbethoxyvinyl; and M+ stands for a cation, e.g. Na+, K+ or (C2H5)3N+H, with eliminating MX, to form - if R2 is hydrogen - the desired compound of formula I. If R2 is a protective group, said group can be removed by methods described in the literature to form the desired compound of formula I.

The reaction is performed in a suitable solvent, e.g. N,N-dimethylformamide, acetone, or hexamethylphosphoric acid triamide, usually at a temperature from about 0 C to about 60"C.

In a second embodiment of the method of the invention, an ester of the formula IV

in which Rl, R2 and X have the above meanings, is reacted with a salt of the formula V

in which M+ and n have the above meanings, to form - if R2 is hydrogen - the desired compound of formula I. If R2 is a protective group said group is removed by methods described in the literature to yield the desired compound of formula I.

The reaction is performed under conditions corresponding to those defined above in the first embodiment of the method according to the invention.

In a third embodiment, a compound of formula VI

in which R1 and X are as defined above, and R2' stands for hydrogen or a protective group, e.g. benzyloxycarbonyl, triphenylmethyl, or ss,ss,ss-trichloroethoxycarbonyl, is reacted with a compound of formula III to yield a compound of formula VII

in which R1, R2 and R2' have the above defined meanings.

By selective removal of the protective group R2,, if present, a compound of formula VIII is obtained:

in which R1 and R2 are as defined above, the said compound is thereafter reacted with a reactive derivative of an amide or thioamide of the formula IX

in which n has the above meaning, and Z stands for oxygen or sulphur, to yield the desired compound of formula I if R2 is hydrogen. If R2 is a protective group, this group is removed by methods described in the literature to yield the desired compound of formula I.

As examples of reactive derivatives of a compound of formula IX, the following non-limiting types may be given:

imrisn chlorides

inn ether

thioamide acetals in which n has the above meaning.

The reactions with the said reactive derivatives are well-known to the man skilled in the art for preparing amidinopenicillanic acid derivatives, confer, e.g., British Patent No.

1,293,590 which also describes in detail the meaning of "a reactive derivative of a compound of formula IX".

In a fourth embodiment, a compound of formula VI is reacted with a compound of formula V to yield a compound of formula X:

in which Rl and R2, have the above defined meanings. If R2, is a protective group, removal of this group yields a compound of formula XI:

in which R1 and n have the above meanings. If R2 is hydrogen, formulae X and XI are identical. The compound of formula X (R2'=H) or XI is thereafter reacted with a reactive derivative of p-hydroxyphenylglycine to yield the desired compound of formula I.

The starting materials of formulae II, III, IV, V and IX are known or may be prepared by methods analogous to those used for the preparation of the known compounds.

The starting material of formula VI can, e.g., be prepared as follows: a salt of 6-amino-penicillanic acid or an N-protected 6-amino-penicillanic acid is reacted with a compound of formula XII:

where R1 is as previously defined and Y is a leaving group, e.g. bromine, iodine or a p-toluenesulphonyloxy group. The reaction is performed in a suitable solvent, e.g.

dimethylformamide or acetone, and usually at a temperature between 0 C and 60"C.

The reaction products of formula I can be purified and isolated in usual manner and may be obtained either in the free state or in the form of a salt.

It is a further object of the present invention to provide pharmaceutical preparations which are useful in the parenteral treatment of infectious diseases in the human and veterinary practice.

With this object in view, the preparations of the invention contain as an active component at least one member selected from compounds of the formula I and salts thereof as defined above, alone or together with an atoxic pharmaceutically acceptable carrier, which may be a solid or liquid and act as a diluent, or solvent and/or together with one or more auxiliary agents.

The preparations can be worked up to various pharmaceutical forms of presentation, suitable for enteral or parenteral administration.

Any known pharmaceutically acceptable, non-toxic organic or inorganic, solid or liquid carrier, diluent, solvent and/or auxiliary agent suitable for parenteral administration can be used to make up preparations containing the present compounds.

The preparations may, if appropriate, contain other therapeutically active components which can be administered together with the present compounds in the treatment of the infectious diseases, such as other antibacterials.

The preparations of the invention can be worked up to any pharmaceutical form suitable for parenteral administration, such as for intravenous, intramuscular or subcutaneous injections. Also the preparation of the invention may be in the form of a pharmaceutical preparation suitable for infusion. Thus the preparation may comprise the active ingredient dissolved or suspended in a non-toxic, pharmaceutically acceptable vehicle. A preparation for injection may typically contain from 5 to 200to by weight of the compounds of the invention, calculated as a compound of formula I. For infusion the preferred content will be in the range of 1% by weight.

Preparations for intramuscular injection may be formulated to produce a sustained release effect, e.g. by giving injections of the active components in the form of suspensions in oil or in other well-known formulations, suitable for such administration.

The compounds of formula I are dibasic and may form salts with one as well as both of the basic moieties. The compounds of formula I are only slightly soluble in water. For use in injection or infusion medicine it is preferred to use a salt of the compound of formula I, e.g.

the dihydrochloride.

Another object of the invention resides in the selection of a dose of the compounds of the invention, which dose can be administered so that the desired activity is achieved without simultaneous secondary effects. In human therapy, the present compounds are conveniently administered (to adults) in dosage units containing not less than 100 mg and up to 2500 mg, preferably from 250 to 1000 mg, calculated as the compound of formula I.Thus a parenteral pharmaceutical preparation in dosage unit form may contain from 100 mg to 2500 mg of at least one compound of formula I or a salt thereof as dry matter in an ampoule, vial or other suitable receptacle for reconstitution, while a pharmaceutical preparation for oral treatment may be in the form of a sustained release preparation in dosage unit form containing at least one compound of formula I in which the dose of the active compound is from 100 mg to 2500 mg.

By the term 11dosage unit" is meant a unitary, i.e. a single, dose which is capable of being administered to a patient, and which may be readily handled and packed, remaining as a physically stable unit dose comprising either the active material as such or a mixture of it with solid or liquid pharmaceutical diluents, carriers, solvents and/or auxiliary agents.

In the form of a dosage unit which may be in the form of a tablet or capsule and which may additionally contain other antibiotics, the compound may be administered one or more times a day at appropriate intervals, always depending, however, on the condition of the patient, and in accordance with the prescription made by the medical practitioner.

Thus a daily dose for a human will preferably be an amount of from 0.5 to 5 g of a compound of formula I or an equivalent amount of a salt thereof as defined before, administered 2 - 4 times daily, which dose is substantially equivalent to 10 to 100 mg per kg body weight.

The preparations of the invention can, as mentioned above, be administered enterally, especially by the oral route.

Still another object of the invention is to provide a method of treating patients suffering from infectious diseases, the method comprising administering to adult non-human patients from 10 to 100 mg per kg body weight per day of at least one compound of the formula I or an equivalent amount of a salt as defined before of a compound of the formula I.

Preferably, the compound is given in the form of the dosage units aforesaid.

The invention will be further described in the following Examples which are not to be construed as limiting the invention.

EXAMPLE 1 6-[(hexahydro-I H-azepin -1 -yl) -methyleneamino]-penicillanoyloxymethyl 6' - (D (-) -p- hydroxy-ct-amino-phenylacetamida) o) -penicillanate and its dihydrochloride To a solution of chloromethyl 6-[(hexahydro-1H-azepin-1-yl)-methyleneamino]- penicillanate (3.73 g, 0.01 M) in dimethylformamide (25 ml), potassium 6-(D(-)-a benzyloxycarbonylamino-p-hydroxy-phenylacetamido)-penicillanate (5.91 g, 0.01 M) was added. The mixture was stirred for one hour and left overnight at 250C. Water (100 ml) and ethyl acetate (100 ml) were added and the organic layer was separated. The aqueous phase was extracted once with ethyl acetate (25 ml) and the combined ethyl acetate extracts were washed ten times with 25 ml portions of McIlvaine buffer (pH 4.0) in order to remove unreacted chloromethyl ester. To the organic phase, water (25 ml) was added and the pH was adjusted to 7.5 with 1 M potassium bicarbonate solution. The organic phase was separated and washed with water (2 x 10 ml). Fresh water was added (50 ml) and pH adjusted to 2.6 with 1 N hydrochloric acid. Palladium catalyst (10% on carbon, 3 g) was added and the mixture was hydrogenated at atmospheric pressure with simultaneous addition of 0.2 N hydrochloric acid to maintain a pH of 2.6. When the consumption of acid stopped (after about 2 h), the catalyst was filtered off. The aqueous layer was separated and residual ethyl acetate was removed in vacuum.After a final filtration the aqueous solution was freeze-dried to yield the dihydrochloride of the title compound as a light yellow powder.

The NMR-spectrum of the product which is indicated in the table is in agreement with the structure.

NMR - SPECTRUM OF: 6-[(hexahydro-1H-azepin-1-yl)-methyleneamino]-penicillanoyl-oxymethyl-6-(D(-)-Phydroxy-α-amino-phenylacetamido)penicillanate, dihydrochloride.

Solvent: CD3OD, TMS ref., # scale p.p.m.

IN EE Ov, EZ V) 3 0S 5,6H) -O-CH2-O- cCH3 {H2CH2 -CH2 N- -CR = N 3' H) 5',6'H) CH2 -CH2 NH H H z II D 4.62, s,1R N S 1.58, s,3R z E = x O | 0 nt fS N g es, g mene / ca c.e V) V) . m 00N00e VR tumb \ H~HH O V) X D O X -sX X vD v) mls t.X SX ~n câ 0 se te The dihydrochloride was dissolved in water, ethyl acetate was added, and a sodium bicarbonate solution was added while stirring until the pH in the aqueous solution was about 7. The organic phase was separated, dried, and evaporated in vacuum leaving the title compound as a yellow oil.

EXAMPLE 2 6-[(1-piperidyl)-methyleneamino]-penicillanoyloxymethyl 6'-(D (-)-p-hydroxy-o- aminophenylacetamido)-penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting chloromethyl 6-[(1-piperidyl)- methyleneamino]-penicillanate for chloromethyl6-(hexahydro 1H-azepin- 1-yl)- methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 3 6-[(hexahydro-1 (2H) -azocin-1 -yl) -methyleneaminol-penicillanoyloxymethyl 6' - (D (-) -p hydroxy-α-aminophenyl-acetamido)-penicillanate and its dihydrochloride.

By following the procedure of Example 1, but substituting chloromethyl 6-[(hexahydro 1(2H)-azocin-1-yl)-rnethyleneamino]-penicillanate for chloromethyl 6-[(hexahydro-1H- azepin-1-yl)-methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 4 6-[(octahydro-1H-azonin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6-(D(-)-phydroxy-a-aminophenyl-acetamido) -penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting chloromethyl 6-[(octahydro1H-azonin-1-yl)-methyleneamino]-penicillanate for chloromethyl 6-(hexahydro-1H- azepin-1-yl)-methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 5 1"- (6-[(hexahydro-1 H-azepin-1 -yl) -methyleneamino]-penicillanoyloxy) -ethyl 6 ' - (D (-) -p- hydroxy-cc-aminophenyl-acetamido) -penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting 1'-chloroethyl 6-[(hexahydro1H-azepin-1-6l)-methyleneamino]-penicillanate for chloromethyl 7-[(hexahydro-1Hazepin-1-yl)-methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 6 1"- (6- [(1 -piperidyl) -methy leneamino]- penicillanoyloxy) -ethyl 6'- (D (-)-p-hydroxy-a- aminophenylacetamido) -penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting 1'-chloroethyl 6-[(1-piperidyl)methyleneamino]-penicillanate for chloromethyl 6-[(hexahydro-1H-azepin-l-yl)- methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 7 1"- (6-J(hexahydro-1 (2H) -azocin-1 -yl) -methyleneamino]-penicillanoyloxy)-ethyl 6'- (D(-)-phydroxy-a-aminophenyl-acetamido)-penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting 1'-chloroethyl 6-[(hexahydro 1 (-2H)-azocin- 1-yl)-methyleneamino]-penicillanate for chloromethyl 6-(hexahydro-1H- azepin-1-yl)-methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 8 1"-(6-[(octahydro-1 H-azonin-I-yl)-methyleneamino]-penicillanoyloxy)-ethyl 6'-(D(-)-p hydroxy-α-aminophenyl-acetamido)-penicillanate and its dihydrochloride By following the procedure of Example 1, but substituting 1'-chloroethyl 6-[(octahydro 1H-azonin-1-yl)-methyleneamino]-penicillanate for chloromethyl 6-[(hexahydro-1H- azepin-1-yl)-methyleneamino]-penicillanate, the desired compound is obtained.

EXAMPLE 9 Preparation for intravenous use 500 g of the (sterilely produced) dihydrochloride of the compound of Example 1 was passed through a vibratlon sieve with 0.10 mm meshes. The resulting product was aseptically dosed into 1000 vials, each thus containing 500 mg of the compound of Example 1.

For injection purposes, the contents of one vial is dissolved in 5 ml of sterilely filtered pyrogen-free water.

For infusion purposes, the contents of one vial is dissolved in 50 ml of an isotonic, sterile saline solution.

EXAMPLE 10 Tablet Ingredients for 1000 tablets: 6-[(hexahydro-1H-azepin-1-yl)-methylene-amino]-penicillanoyloxymethyl 6'-(D(-)-p hydroxy-α-amino-phenylacetamiddo)-penicillanate, dihydrochloride (A) .. .. 250 g Polyvinylpyrrolidone . ...... ....... . . ..... .................... ......... .... 10 g Isopropanol .. . ....... . . ....... ....... . ....................... 100 ml Microcrystalline cellulose . ....... ....... . 165 g Corn starch ... .. . . . ...... . . 71 g Magnessium stearate . .................................. . . 4 g Sieve A through a screen with 1 mm openings and wet the powder with a solution of polyvinylpyrrolidone in isopropanol. Dry the moist mass at 300C and pass it through a sieve with 0.7 mm openings.

Mix the granules with microcrystalline cellulose, corn starch and magnesium stearate.

Press tablets of 0.500 g weight using 12 mm punches and dies to yield tablets each containing 250 mg of A.

EXAMPLE 11 6-[(Hexahydro-1H-azepin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6'-(D(-)-p hydroxy-α-amino-phenylacetamido)-penicillanate, dihydrochloride A. 6-[(Hexahydro-1H-azepin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6'benzyloxycarbonylamino-penicillanate, hydrochloride To a solution of chloromethyl 6-[(hexahydro-1H-azepin-1-yl)-methyleneamino]penicillanate (5.75 g) in N,N-dimethylformamide (60 ml), potassium 6 benzyloxycarbonylamino-penicillanate (7.8 g) was added. The mixture was stirred for 40 hours at room temperature. Then, ethyl acetate (250 ml) was added and the mixture extracted with water (3 x 50 ml), saturated aqueous sodium bicarbonate (50 ml) and water (50 ml). To the organic phase, water (200 ml) was added and the pH-value of the aqueous phase was adjusted to 2.5 by addition of 4 N hydrochloric acid.The aqueous phase was separated, filtered and freeze-dried to yield the desired compound as an amorphous powder.

The NMR-spectrum (CD3OD) showed signals at 5 = 1.53(s); 1.57(s); 1.65(s); 1.73(s); 1.4-2.1(m); 3.4-4.0(m); 4.49(s); 4.60(s); 5.13(s); 5.3-5.7(m); 5.95(s); 7.34(s); 8.18(s) ppm.

TMS was used as internal reference.

B. 6-[(Hexahydro-1H-azepin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6'aminopenicillanate, dihydrochloroide To a stirred solution of the compound prepared in step A (2.0 g) in water (100 ml), ethyl acetate (50 ml) and hydrochloric acid to pH=3 was added. Catalyst (10% Pd on carbon, 2.0 g) was added and hydrogen was bubbled through the stirred mixture, a pH-value of 3 being maintained by the addition of 1 N hydrochloric acid. When the consumption of acid ceased, the catalyst was filtered off, and the aqueous phase was separated and freeze-dried to yield the desired compound as an amorphous powder.

The NMR-spectrum (CD3OD) showed signals at 5 = 1.58(s); 1.74(s); 1.5-2.2(m); 3.5-4.0(m); 4.62(s); 4.67(s); 5.08(m); 5.5-5.8(m); 5.97(s); 8.23(s) ppm. TMS was used as internal reference.

C. 6-[(Hexahydro-1H-azepin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6'-(D(-) p-hydroxy-α-amino-phenylacetamido)-penicillanate, dihydrochloride To a mixture of D(-)-N-benzyloxycarbonyl-p-hydroxyphenylglycine (3.01 g), triethylamine (1.4 ml) and acetone (100 ml), cooled to --200C, isobutyl chloroformate (1.7 ml) was added and the mixture was stirred for 30 minutes at -20 C to form the mixed anhydride.

An ice-cooled solution of 6-[(hexahydro-1H-azepin-1-yl)-methyleneamino]penicillanoyloxymethyl 6'-amino-penicillanate (liberated from the dihydrochloride prepared in step B, 5.5 g) in acetone (50 ml) was added to the mixed anhydride. The reaction mixture was stirred for 1 hour at room temperature and evaporated in vacuo to half the initial volume. Ethyl acetate (200 ml) was added and the mixture was extracted with water (2 x 50 ml), saturated aqueous sodium bicarbonate (50 ml), and water (50 ml). Then water (200 ml) was added and the pH adjusted to 2.6 with 1 N hydrochloric acid. The aqueous phase was isolated, 10% palladium on carbon catalyst (3 g) was added, and the mixture was hydrogenated as described in Example 12, step B to yield a product the NMR-spectrum of which was identical with that described in Example 1.

EXAMPLE 12 6-(Hexahydro-1H-azepin-1-yl)-methyleneamino]-penidllanoyloxymethyl 6'-(D(-)-p- hydroxy-a-amino-phenylacetamido)-penicillanate, dihydrochloride A. Chloromethyl 6- (D (-) -a-benzyloxycarbonylamino-p-hydroxy-phenylacetamido) penicillanate To a suspension of potassium 6-(D(-)-a-benzyloxycarbonylamino-p-hydroxyphenylacetamido)-penicillanate (7 g) in N,N-dimethylformamide (25 ml), chloroiodomethane (8 ml) was added, and the mixture was stirred for 4 hours at room temperature. Then, the mixture was diluted with ethyl acetate (100 ml) and washed with water (3 x 25 ml), saturated aqueous sodium bicarbonate (10 ml) and water (2 x 10 ml).

The organic phase was dried and evaporated in vacuo to yield the crude ester as a brownish oil, which could be used in the next step without further purification.

B. 6-[(Hexahydro-1 H-azepin -1 -yl) -methyleneamino]-penicillanoyloxymethyl 6' - (D (-) p-hydroxy-a-aminoph enylacetamido) -p en icillanate, dihydrochloride To a solution of chloromethyl 6-(D(-)-a-benzyloxycarbonylamino-p-hydroxyphenylacetamido)-penicillanate (6.01 g) in N,N-dimethylformamide (25 ml), sodium 6-[(hexahydro-1R-azepin-1-yl)-methyleneaminoj-penicillanate (3.47 g) was added, and the mixture was stirred for 40 hours at room temperature. The mixture was diluted with ethyl acetate (100 ml) and extracted with water (3 x 25 ml). To the organic phase, water (200 ml) was added and the pH adjusted to 2.5 with 1 N hydrochloric acid. The aqueous phase was isolated and washed with diethyl ether (25 ml).Then catalyst (10% palladium on carbon, 3 g) was added and hydrogen bubbled through the solution, a pH-value of 2.6 being maintained by addition of 1 N hydrochloric acid. When the consumption of acid stopped, the catalyst was filtered off and the filtrate was freeze-dried to give a product the NMR-spectrum of which was identical with that described in Example 1.

EXAMPLE 13 6-[(Hexahydro-lH-azepin-1-yl)-methyleneamino]-penicillanoyloxymethyl 6'-(D(-)-phydroxy-a-amino-phenylacetamido)-penicillanate, dihydrochlorode A. 6-(D(-)-q-Benzyloxycarbonylamino-p-hydroxy-phenyl-acetamido)- penicillanoyloxymethyl 6"-aminopenicillanate To a mixture of D(-)-N-benzyloxycarbonyl-p-hydroxyphenylglycine (3.01 g), triethylamine (1.4 ml) and acetone (100 ml), cooled to -200C, isobutyl chloroformate (1.7 ml) was added and the mixture was stirred for 30 minutes at -200C to form the mixed anhydride.

A solution of bis(6-aminopenicillanoyloxy)-methane (4.44 g) in acetone (50 ml) was cooled to -200C and added to the mixed anhydride in one portion. The reaction mixture was stirred for 2 hours while the temperature was allowed to rise to room temperature.

Ethyl acetate (300 ml) was added and the mixture was extracted with water (2 x 50 ml), saturated aqueous sodium bicarbonate (50 ml) and water (50 ml). The organic phase was dried and evaporated in vacuo to leave an oil, which after purification by dry column chromotagraphy on silica gel, yielded the desired compound as a yellow gum.

B. 6-[(Hexahydro-lH-azepin-1 -yl) -methyThneamino]-penicillanoyloxymethyl 6' - (D (-) - p-hydroxy-a-amino-phenylacetamido)-penicillanate, dihydrochloride To a solution of the compound prepared in step A, (6.3 g) and N,Ndiisopropylethylamine (1.7 ml) in dry chloroform (70 ml) at OOC, Nformylhexamethyleneimine dimethyl sulphate complex (2.5 g) was added, and the reaction mixture was stirred for 20 hours at room temperature. The solution was evaporated, the residue was taken up in diethyl ether (300 ml) and the precipitate was filtered off. Water (200 ml) was added and the apparent pH-value was adjusted to 2.5 by addition of 1 N hydrochloric acid. The aqueous phase was separated, 10% palladium on carbon catalyst (5.0 g) was added, and the mixture was hydrogenated as described in Example 12 to yield a product the NMR-spectrum of which was identical with that described in Example 1.

Claims

WHAT WE CLAIM IS:

1. A compound of the formula I:

in which the asterisk indicates an asymmetric carbon atom and "+" the possibility of an asymmetric carbon atom, and R, represents hydrogen or methyl, and n is an integer from 5 to 8, and salts thereof with pharmaceutically acceptable non-toxic acids.

2. A compound according to claim 1, in which the grouping

represents 1-piperidyl, hexahydro-lH-azepin-1-yl, hexahydro-1(2H)-azocin-1-yl, or octahydro-lH-azonin-1-yl, and R1 represents hydrogen.

3. 6-[(hexahydro-1H-azepin- 1-yl)-methyleneamino]-penicillanoyloxymethyl 6' -(D(-)- p-hydroxy-a-amino-phenylacetamido)-penicillanate and its dihydrochloride.

4. Method for producing a compound of formula I of claim 1 in which a) an ester of the formula II:

in which formula R1 and n are as defined above, and X is a leaving group is reacted with a salt of the formula III:

in which R2 represents hydrogen or a protective group, and M+ stands for a cation, with the elimination of MX, to form, if R2 is hydrogen, the desired compound of formula I, and, if R2 is a protective group, a compound, from which said group is removed by methods described in the literature to form the desired compound of formula I; or b) - an ester of the formula IV:

in which R1, R2 and X have the above meanings, is reacted with a salt of the formula V:

in which M+ and n have the above meanings, to form, if R2 is hydrogen, the desired compound of formula I, and, if R2 is a protective group, a compound, from which said group is removed by methods described in the literature to yield the desired compound of formula I; or c) a compound of formula VI:

in which R1 and X are as defined above, and R2, stands for hydrogen or a protective group, is reacted with a compound of formula III to yield a compound of formula VII:

in which R1, R2 and R2, have the above defined meanings, whereafter by selective removal of the protective group R2,, if present, a compound of formula VIII is obtained:

in which R1 and R2 are as defined above, and said compound is thereafter reacted with a reactive derivative of an amide or thioamide of the formula IX:

in which n has the above meaning, and Z stands for oxygen or sulphur, to yield the desired compound of formula I if R2 is hydrogen, and, if R2 is a protective group, this group is removed by methods described in the literature to yield the desired compound of formula I; or d) a compound of formula VI is reacted with a compound of formula V to yield a compound of formula X:

in which Rl and R2, have the above defined meanings, and, if R2' is a protective group, removal of this group yields a compound of formula XI:

in which Rl and n have the above meanings, and whereafter the compound of formula X (R2, = H) or XI is reacted with a reactive derivative of p-hydroxyphenylglycine to yield the desired compound of formula I.

5. A method as claimed in claim 4(a) wherein X is chlorine, bromine, iodine or p-toluenesulphonyloxy, M is a sodium or potassium ion or (C2H5)3 N+H and the protective group, if present, is benzyloxycarbonyl, triphenylmethyl ss,ss,ss-trichloroethoxycarbonyl, or 1-methyl-2-carbethoxyvinyl.

6. A pharmaceutical preparation in dosage unit form for the enteral or parenteral treatment of patients suffering from infectious diseases, which comprises as an active ingredient at least one compound of formula I, or a pharmaceutically acceptable, non-toxic salt thereof, and an atoxic pharmaceutically acceptable carrier, the quantity of the said active compound being between 100 mg and 2500 mg.

7. A preparation as claimed in claim 6 wherein the dosage unit contains from 250 mg to 1000 mg of at least one compound of formula I or a salt thereof as defined in claim 1.

8. A preparation as claimed in claim 6 or 7 wherein the dosage unit is in the form of a tablet.

9. A preparation as claimed in claim 6 or 7 wherein the dosage unit is in the form of a capsule.

10. A parenteral pharmaceutical preparation in dosage unit form, containing from 100 mg to 2500 mg of at least one compound of formula I of claim 1 or a salt thereof as defined in claim 1 as dry matter, in an ampoule, vial or other suitable receptacle, for reconstruction.

11. A preparation according to claim 6 wherein the active inredient is dissolved or suspended in a non-toxic, pharmaceutically acceptable vehicle.

12. A pharmaceutical preparation for oral treatment in form of a sustained-release preparation in dosage unit form of at least one compound of claim 1, in which the dose of the active compound is from 100 mg to 2500 mg.

13. A preparation as claimed in any one of claims 6 to 12 in which the dosage unit additionally contains other antibiotics.

14. A method of treating non-human patients suffering from infectious diseases, comprising administering to (adult) patients from 10 to 100 mg per kg body weight per day of at least one compound of formula I of claim 1 or an equivalent amount of a salt thereof as defined in claim 1.

15. A method according to claim 14, in which the compound of formula I or salt thereof is administered in the form of a preparation according to any one of the claims 6 to 13.

16. A compound of the formula I defined in claim 1 substantially as hereinbefore described in any one of the foregoing Examples.

17. A method for producing a compound of formula I defined in claim 1 substantially as hereinbefore described in any one of Examples 1 to 8 and 11 to 13 of the foregoing Examples.