DE2726208A1 - Protected di:O-amino:glycosyl-di:amino:cyclitol derivs. - which are selectively acylated or sulphenylated and their use in antibiotic prodn. - Google Patents

Abstract

Selectively acylated for sulphenylated 4,6-di-O-(aminoglycosyl)-1,3-diaminocyclitols of formula (I) are new with substits. as follows: (a) E is CH2NHR1 or CHMeNHR1; F, G and J are OH; (b) E is CH2NHR1, CHMeNHR1, CHMeNMeR1 or CH2NMeR1 and J is NHR2; (c) E is CH2OH, CH2NHR1, CHMeNHR2 or CHMeOH; F and G are OH and J is NHR2; (d) E is CH2NHR1; F is OH and J is NHR2; (e) K is CH2NHR1, CHMeNHR1 or CH2NMeR1; (f) M is OH; Q is Me and T is NMER5; (g) L is CH2OH; Q is OH and T is NHR5; (h) M is OH and T is NMeR5; (i) Q is OH and T is NMeR5; substits. not specified are H; U, V and W are H or OH; Z is H, OH or amino; R1-5 are H, -SR1 or -CO-A, 1-4 of the R1-5 gps. being H; R1 is opt. substd. phenyl, di- or tri-phenylmethyl; A is -(CH2)n-B or -O-C (CH2)nD 3; B is H or opt. substd. phenyl; D is H or one D can be opt. substd. phenyl; n = 0-5 independently for each n. (I) are intermediates for derivs. of 4,6-di-O-(aminoglycosyl)-1,3-diaminocyclitols, esp. derivs. of the antibiotics Gentamicins A, B, B1, C1, C1a, C2, C2a, C2b and X2, Siso-micin, Verdamicin, Tobramycin, Antibiotics G-418, 66-40B, 66-40D, JI-20A, JI-20B and G-52 and Mutamicins 1, 2, 4, 5 and 6. These antibiotics are effective against bacterial infections but frequently have relatively high nephro- and ototoxicity and can lead to the development of resistant bacteria. These problems can be overcome by the use of derivs., e.g. 1-N-acetyl- and 1-N-ethyl-sisomicin, which can be produced more easily from (I) than from unprotected sisomicin.

Description

1-N-formylsisomicin, process for its preparation as well as

Its use as a medicament The present invention relates to 1-N-formylsisomicin, process for its preparation and its use as a medicinal product, especially as an antimicrobial agent.

Aminoglycoside antibiotics are important substances to be effective Fight bacterial infections. However, the occurrence of resistant germs is reduced in many cases their broad applicability; furthermore, side effects such as about ototoxicity and nephrotoxicity occur. Derivatisation works in some of them Cases to avoid these disadvantages.

Such derivatives of aminoglycoside antibiotics are already known are e.g.

1-N- (4-amino-3-hydroxy) -butyrylkanamycin A, 1 -N-acetylsisomicin and 1-N-ethylsisomicin (DT-OS 2,437,160).

It has now been found that 1-N-formylsisomicin (1) and its pharmaceutically acceptable salts have strong antibacterial properties against a large number of germs.

The pharmaceutically acceptable salts are derived from inorganic or organic acids such as sulfuric, phosphoric, nitric, hydrochloric, hydrobromic, acetic, propionic, ascorbic, citric acids, etc. from. To prepare 1-N-formylsisomicin, the general procedure is that a compound of the formula II in which R1, R2, R3, R4 denote hydrogen or easily removable amine protective groups, or their salts, which contain an inorganic or organic acid radical, are converted into the IN-formyl compound in a manner known per se by reaction with a formylating agent, the protective groups optionally in an splits off in a known manner and works up the approach to 1-N-formylsisomicin.

The 1-N-formyl compound can be used during work-up or after it then by reaction with suitable acids into their pharmaceutically acceptable salts convict.

Easily split off protecting groups are to be understood as meaning those which are used in peptide syntheses to protect the amino groups and through alkaline or acidic hydrolysis, gentle hydrogenolysis or nucleophilic displacement reactions with retention of the introduced l-N-formyl group and without influencing the sisomicin structure can be removed again. A large number of such protective groups are known (Houben-Weyl, Methods of Organic Chemistry, Volume XV 1, Pages 46-305, Georg Thieme Verlag, Stuttgart, 1974). According to formula II, one or all of the N atoms 3, 2 ', 6 ', 3 "; it is also possible to start from free sisomicin.

The following radicals may be mentioned by way of example as N-protecting groups: o-tlitrophenylsulfonyl, carbobenzoxy, phthaloyl, t-butoxycarbonyl, trifluoroacetyl, Tosyl, p-nitrocarbobenzoxy and cyclopentyloxycarbonyl.

A compound of the formula II is expediently used for the formylation in the presence of a diluent, organic solvents are suitable as such such as.

Dichloromethane, toluene, dimethylformamide, dimethyl sulfoxide, pyridine as well as their mixtures with water and / or methanol - with expediently at least 1 mole of a formylating agent, with appropriate blocking of the amine functions an up to 10-fold excess Ve, enduna 'can be found, if necessary in presence an organic or inorganic base as an acid scavenger to.

Particularly suitable formylating agents are p-nitrophenyl formates as well as the mixed anhydride of formic and acetic acid. In principle, however any other reactive formyl derivative can also be used.

The reaction temperatures can vary within a wide range will. In general, temperatures of about -300 ° C. to + 500 ° C. are used, preferably between about OOC and about + 250C.

The production of 1-N-formylsisomicin is particularly advantageous, if you have 2 ', 3, 3 ", 6'-Tetra-N- (6-nitrophenylsulfenyl) -sisomicin, which after a new process (Le A 18 047 - German patent application p by sulfenylation of Sisomicin or its SE.ureadditionsalzen is accessible, with about one to about 5 moles of formylating agent converts the o-nitrophenylsulfenyl groups with sulfur-containing, nucleophilic reagents such as H2S or thiophenol are split off and the reaction mixture worked up in the usual way on the free-N-formyl derivative and this optionally converted into the pharmaceutically acceptable salts.

The sulfenyl derivative is preferably formylated in pyridine as the diluent. If p-nitrophenyl formate is used as the formylating agent in the reaction described above, the course of the reaction can be represented by the following equation: Instead of the o-nitrophenylsulfenyl derivatives, other sulfenyl derivatives which can be obtained in an analogous manner by reacting sisomicin with corresponding sulfonic acid chlorides or the sulfonic acid p-nitrophenyl esters can also be used with good success. Examples are: tritylsulfenyl, pentachlorophenylsulfenyl, 2,4-dinitrophenylsulfenyl-sisomicin derivatives.

Another display option is the direct implementation of Sisomicin or its salts of inorganic or organic acids with one mole Formylating agent in a manner known per se and subsequent chromatographic Separation of the desired 1-N-formyl derivative.

Surprisingly, show the 1-N-formylsisomicin according to the invention as well as its pharmaceutically acceptable salts with good tolerability a higher one Effect against resistant bacterial strains than those known from the prior art Verbirdllngon.

The substances according to the invention are thus an enrichment for pharmacy represent.

The compounds of the invention are antimicrobial agents with a broad spectrum of activity and particular effectiveness against gram-negative bacteria. These properties allow them to be used as drugs in combating of bacterial diseases in humans and animals.

They are good for prophylaxis and chemotherapy of local and systemic Infections, especially infections of the genitourinary system in human and veterinary medicine suitable that are caused by gram-negative bacteria, e.g. E. coli, Proteus, Klebsiella and Pseudomonas are suitable. Inhibition depths in the A gar hole test were e.g. against the following bacterial strains at a concentration of 100 micrograms / 1 ml found; Pseudomonas aerug. 5737 Pseudomonas aerug. F 41 Klebsiella pneum. 2 Munich Klebsiella pneum. 1 Düsseldorf E. coli Munster E. coli Neumann In general, depends the dose to be administered of the compounds of the invention on the age and weight of the living being, the method of administration, the type and severity of the bacterial Infection. The dosage of the compounds of the invention is usually that Dosage of the 1-N-unsubstituted compounds similar. The dosage range is from 20 mg / day / human to 2000 / mg / day / human, preferably 100 mg - 500 mg / day.

The compounds of the invention can be administered orally.

The administration can be divided into single doses or in several doses take place. They can also be administered topically in the form of ointments, creams or lotions. Pharmaceutical carriers for these formulations include water, oils, fats, polyesters and polyols.

For oral administration of the compound of this invention, tablets, Capsules or elixirs are used, the connections can but can also be mixed with animal feed.

In general, topical preparations contain from about 0.1 to about 3.0 g of the compounds of the invention per 100 g of ointment, cream or lotion. The topical Administration takes place approx.

2 to 5 times a day.

The antibacterial agents of the invention can be in liquid form as solutions or suspensions for use on the ears and eyes or for parenteral use Administration in the form of intramuscular injections. Injection solutions or suspensions are usually administered so that approx.

1 to 15 mg of active ingredient per kilogram of body weight in 2 to 4 doses get into the infected organism per day.

The exact dose depends on the type of infection and sensitivity the infecting germ and the individual characteristics of the person to be treated away.

Formulation 1 tablet 10 mg tab. 25 mg tab. 100 mg tab.

1-N-formyl- + + 26.2 '+ + sisomicin 10.50 mg 26.25 mg 105.00 mg lactose 197.50 mg 171.25 mg 126.00 mg corn starch 25.00 mg 25.00 mg 35.00 mg polyvinylpyrrolidone 7.50 mg 7.50 mg 7.50 mg Magnesium stearate 2.50 mg 2.50 mg 3.50 mg + 5% excess Manufacturing It becomes a slurry of 1-N-formylsisomicin, lactose and polyvinylpyrrolidone and these are spray-dried. You give the corn starch and the magnesium stearate to, mixes and compresses into tablets.

Formulation 2 Ointment 1-N-Formylsisomicin 1.0 g methyl paraben U.S.P. 0.5 g propyl paraben U.S.P. 0.1 g petrolatum per 1000 g Preparation (1) Melt the petrolatum; (2) Mix 1-N-formylsisomicin, methyl paraben and propyl paraben with about 10% of the melted petrolatum; (3) Place the mixture in a colloid mill; (4) Add the remaining petrolatum while stirring and cool until it becomes semi-solid. The product is filled into suitable containers.

Formulation 3 solution for injection Per 2.0 ml vial per 50 liters of 1-N-formylsisomicin 84.0 mg 2100.0 gm methyl paraben, U.S.P. 3.6 mg 90.0 gm propyl paraben, U.S.P. 0.4 10.0 gm sodium bisulfite, U.S.P. 6.4 mg 160.0 gm disodium ethylenediamine 0.2 mg 5.0 gm tetraacetate dihydrate water, U.S.P. q.s. 2.0 mg 50.0 liters 5% excess example 1 a) Penta-N- (o-nitrophenslsulfenyl) -sisomicin 13.84 g (20 m M) sisomicin sulfate in 100 ml of 1 N NaOH and 450 ml of freshly distilled dioxane are mixed with 38 g (0.2 M) o-Nitrophenylsulfenyl chloride in 200 ml of dioxane and added with 260 ml of 1 N NaOH so that that a pH of 12-14 is established. The precipitate is filtered off, in dissolved in a CH2Cl2-water mixture and the dichloromethane phase dried over Na2S04.

Dichloromethane is added to the filtrate and the aqueous phase is discarded and the dichloromethane extract dried over Na2S04. The united organic Phases are evaporated to dryness and passed through a column with a diameter of 8 cm, which contains 250 g of silica gel, first with dichloromethane, then with a Dichloromethane-Methal mixture (97.5 / 2.5) filtered. The filtrate yields after evaporation of the solvent 22 g (91%) of penta-N- (o-nitrophenylsulfenyl) sisomicin as an orange Foam.

b) 3 "-N- (o-nitrophenylsulfonyl) lsisomicin 16.0 g (13.2 m M) des under a) produced penta-N- (O-nitrophenylsulfenyl) -sisomicin in 80 ml absol. 160 ml of thiophenol are added to pyridine, after 1 hour to 500 ml of diethyl ether poured, the precipitate in dichloromethane / methanol (8/2) and taken over Filtered silica gel (column 5.5 x 12 cm, mobile phase dichloromethane / methanol = 8/2, increasing addition of the solvent mixture methanol / dichloromethane / 20 percent. ammonia = 4/2/1). After evaporation of the solvent, the red zone yields 6.6 g (83 %) 3 "-N-o-nitrophenylsulfenylsisomicin as a deep red foam.

c) 2'e3t3 ||| 6 | -Tetra-N- (O-nitrophenylsulfenyl) -sisomicin 3.0 g (5.0 m M) of the 3 "-N-o-nitrophenylsulfenyl-sisomicin prepared under b) in 5 ml of methanol and 45 ml of dichloromethane are mixed with 4.4 g (15.0 m M) of o-nitrophenylsulfenic acid p-nitrophenyl ester added in 85 ml of dichloromethane, the reaction mixture immediately evaporated to dryness, taken up in dichloromethane and on silica gel (column 5.5 x 30 cm) with 200 ml of dichloromethane, then chromatographed with dichloromethane / methanol (98/2). There will be 500 parliamentary groups collected, whereby from the combined fractions 270 to 500 the 2 ', 3.3 ", 6'-Tetra-N- (o-nitro-phenylsulfenyl) -sisomicin is obtained as an orange foam.

d) 1-N-Formylsisomicin 265 mg of the 2 ', 3.3 ", 6'-tetra- (N-o-nitrophenylsulfenyl) -sisomicin obtained according to c) and 44 mg of p-nitrophenyl formate are dissolved in 1 ml of pyridine. One lets Stand at room temperature for 1 hour, evaporate the batch in vacuo and take in 2 ml of dichloromethane. The solution is 7.5 ml of a methanolic solution which is saturated at 0 ° C H2S solution and 0.75 ml of a methanolic hydrogen chloride solution saturated at 200C added and after 1 minute with conc.

Ammonia solution neutralized. The batch is evaporated in a vacuum, digested the residue with 10 ml of water and filtered. The filtrate is made twice Washed with ether, the aqueous phase in 12 ml of basic ion exchanger in the OH form filtered and evaporated.

Yield: 102 mg of 1-N-formylsisomicin IR (KBr pellet) -1 band at 1663 cm

Claims (6)

Claims l.l-N-formylsisomicin and its pharmaceutical usable salts. i2. Process for the preparation of 1-N-formylsisomicin and its pharmaceutically acceptable salts, characterized in that a compound of the formula II in which R1, R2, R3, R3 are hydrogen or easily removable amine protective groups, or their salts, which contain an inorganic or organic acid radical, are converted into the 1-N-formyl compound in a manner known per se by reaction with a formylating agent, the protective groups optionally split off in a manner known per se, worked up the batch to 1-N-formylsisomicin and optionally converted into a pharmaceutically usable salt. 3. The method according to claim 1, characterized in that 2 ' , 3,3 ", 6-tetra- <o-nitro-phenylsulfenyl) -sisomicin with about 1 to about 5 mol a formylating agent, preferably Formic acid p-nitrophenyl ester, or formic acid-acetic acid-anhydride, in an organic solvent, after the reaction has ended, the o-nitrophenylsulfenyl protective groups with a nucleophilic Reaquenz, preferably hydrogen sulfide solution, or thiophenol split off and then the reaction mixture for the formyl compound or one of its pharmaceutically acceptable compounds Salts worked up. 4. Medicinal products characterized by a content of a compound according to claim 1. 5. Process for the production of medicaments according to claim 4, characterized in that characterized in that a compound according to claim 1 with pharmaceutically suitable Mixed carriers. 6. Use of compounds according to Claim 1 in combating of diseases in humans and animals.