DE10064778A1 - New 3-substituted 6-anilino-uracil derivatives, useful as antibacterial agents with high solubility and good activity against Gram positive bacteria - Google Patents

Abstract

3-Substituted 6-anilino-uracil derivatives (I) are new. Amino-uracils of formula (I) and their salts, solvates and hydrates are new. R1 = cycloalkyl, cycloalkenyl, 6-10C aryl or 4-10 membered, optionally fused heteroaryl containing 1-4 of O, S and N as heteroatom(s), all optionally substituted by 1-3 of halo, CF3, OCF3, NO2, NH2, mono- or dialkylamino, acylamino and oxo; R2 = 4-(R2a)-3-(R2b)-phenyl; R2a, R2b = alkyl or halo; or R2a + R2b = group completing a 5- or 6-membered ring (optionally containing 1 or 2 O and optionally substituted by 1-3 halo); A = (CH2)nL(CH2)m; n, m = 0-5; L = S, SO, SO2, CH(OH), CO, C(=NR10) or a direct bond; R10 = H or alkyl. Provided: (1) A is not CH2 if R1 = Ph; and (2) L is not CO or SO2 if R1 = N-bonded ring and n = 0. Independent claims are included for: (1) the preparation of (I); and (2) new chloro-uracil intermediates of formula (II).

Description

The present invention relates to cyclically substituted phenylamino-uracile, Ver proceed to their manufacture, they comprise pharmaceutical compositions as well as their use in the treatment of diseases in humans or Animals.

Gram-positive eubacteria contain three different DNA polymerase exo Nucleases, which are referred to as Pol 1, Pol 2 and Pol 3. Pole 3 is an enzyme which is necessary for the replicative synthesis of the DNA.

WO 96/06614 describes 3-substituted uracils of the following formula:

wherein
R ¹ (CH ₂ ) _n OH, CH ₂ CHOHCH ₂ OH, CH ₂ CHOHCH ₃ , (CH ₂ ) mCO ₂ H, or (CH ₂ ) _n NH ₂ .

WO 00/20556 describes Uracile with a zinc finger active unit as an antibac serial connections.

J. of Med. Chem. 1977, Vol. 20, No. 9, 1186 describes 3-unsubstituted uraciles, also J. of Med. Chem. 1999, Vol. 42. No. 11, 2035.

The suitability of uraciles as antibiotics has long been known (Biochem. et. Biophys. Acta. 281 (1972)).  

Antimicrobial Agents and Chemotherapy, August 2000, 2217-2221, Vol. 44, No. 8 describes 3-alkoxy-alkylidene-substituted uracils.

Antimicrobial Agents and Chemotherapy, August 1999, 1982-1987, Vol. 43, No. 8 describes 6 - ([3,4-trimethylene] anilino) uracil (TMAU), u. a. is substituted benzyl ized TMAU described as practically ineffective against polymerase 3.

One problem with the prior art connections is sufficient To achieve solubility with good effectiveness.

An object of the present invention is to provide new, alternative alkoxycarbonyl alkylidene-uracile with antibacterial effect for the treatment of diseases To make people and animals available.

The present invention therefore relates to compounds of the general formula (I)

wherein
R ¹ is cycloalkyl, cycloalkenyl, C ₆ -C ₁₀ aryl, a 4- to 10-membered, optionally fused heterocycle with up to 4 heteroatoms selected from the group O, S, N, where R ^{1 is} optionally substituted can with 1 to 3 substituents selected from the group halogen, trifluoromethyl, trifluoromethoxy, nitro, amino, alkylamino, dialkylamino, acylamino, oxo,
R ^{2 is} a substituent of the following formula

wherein
R ^2-1 and R ^{2-2 are} independently selected from the group alkyl or halogen, or
R ^2-1 and R ^2-2 together form a 5- to 6-membered ring which can optionally have up to 2 oxygen atoms and which can optionally be substituted by up to 3 halogen,
A a group

wherein
n is a number from 0 to 5,
m is a number from 0 to 5,
L is -O-, -S-, -S (O) -, -S (O) ₂ -, -C (H) OH-, -C (O) -, -C = NR ¹⁰ - or a bond is
where R ¹⁰ is hydrogen, alkyl,
with the proviso that A is not CH ₂ when R ^{1 is} phenyl, and
that L is not equal to -C (O) - or -S (O) ₂ - if R ^{1 is} a ring bonded via a ring nitrogen atom and n is 0,
and their pharmaceutically acceptable salts, solvates and hydrates.

The compounds of general formula (I) according to the invention can in Different stereoisomeric forms occur, which are either like picture and Mirror image (enantiomers), or which are not like image and mirror image (Diastereomers) behave. The invention relates to both the enantiomers and Diastereomers and their respective mixtures. The racemic forms can be as well as the diastereomers in a known manner in the stereoisomerically uniform Separate components.

Furthermore, certain compounds can exist in tautomeric forms. This is known to those skilled in the art, and such compounds are also within the scope of Invention includes.

The substances of the general formula (I) according to the invention can also be used as salts available. In the context of the invention, physiologically acceptable salts are in the offing Trains t.

Physiologically acceptable (pharmaceutically acceptable) salts can be salts of compounds according to the invention with inorganic or organic acids. Salts with inorganic acids such as chlorinated water are preferred acidic acid, hydrobromic acid, phosphoric acid or sulfuric acid, or salts with organic carboxylic or sulfonic acids such as acetic acid, propionic acid, Maleic acid, fumaric acid, malic acid, citric acid, tartaric acid, lactic acid, Benzoic acid, or methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluene sulfonic acid or naphthalene disulfonic acid.  

Physiologically acceptable salts can also be salts of the invention Be compounds with bases, such as metal or ammonium salts. Preferred examples are alkali metal salts (e.g. sodium or potassium salts), earth alkali salts (e.g. magnesium or calcium salts), as well as ammonium salts, the abge are guided by ammonia or organic amines, such as ethylamine, Di- or triethylamine, ethyldiisopropylamine, monoethanolamine, di- or tri ethanolamine, dicyclohexylamine, dimethylaminoethanol, dibenzylamine, N-methyl morpholine, dihydroabietylamine, 1-ephenamine, methylpiperidine, arginine, lysine, Ethylenediamine or 2-phenylethylamine.

The compounds according to the invention can also be in the form of their solvates, in particular which are in the form of their hydrates.

The compounds of the present invention have a broad activity spectrum against Gram-positive bacteria, also multi-resistant Germs can be detected, especially staphylococci, pneumococci and Enterococci, including strains resistant to vancomycin.

The compounds generally have sufficient solubility to To produce solutions for parenteral administration.

Alkyl and the alkyl parts in alkoxy, alkylthio, mono- and dialkylamino, alkyl sulfonyl include z. B. C ₁ -C ₂₄ -, preferably C ₁ -C ₁₂ - and C ₇ -C ₂₄ -, in particular C ₁ - C ₆ - and C ₁ -C ₄ alkyl.

Cycloalkyl comprises polycyclic saturated hydrocarbon radicals with up to 14 carbon atoms, namely monocyclic C ₃ -C ₁₂ -, preferably C ₃ -C ₈ alkyl, in particular special C ₅ -C ₈ alkyl such as. B. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and polycyclic alkyl, ie preferably bicyclic and tricyclic, optionally spirocyclic C ₇ -C ₁₄ alkyl, such as. B. Bicyclo [2.2.1] -hept-1-yl, Bicyclo [2.2.1] -hept-2-yl, Bicyclo [2.2.1] -hept-7-yl, Bicyclo- [2.2.2] -oct -2-yl, bicyclo [3.2.1] oct-2-yl, bicyclo [3.2.2] non-2-yl and adamantyl.

Cycloalkenyl with up to 12 C atoms comprises mono- or polyunsaturated, non-aromatic, mono- or polycyclic alkyl such as cyclopentenyl, cyclo hexenyl, cyclohexadienyl or cyclooctenyl.

A 4- to 10-membered, optionally fused heterocycle with up to 4 hetero atoms selected from the group O, S and N stands for a mono- or bicyc matic heterocycle containing one or more double bonds or aroma can be one to four identical or different heteroatoms from the series N, O and contains S and which ver via a ring carbon atom or a ring nitrogen atom is knotting. Examples include: pyridyl, 1,2-dihydropyridinyl, 1,4-dihydro pyridinyl, furyl, thienyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, triazolyl, pyra zolyl, pyrazinyl, 3,4,5,6-tetrahydropyrimidinyl, pyrimidinyl, pyridazinyl, indolyl, Benzimidazolyl, Benzo [b] thienyl, Benzo [b] furyl, Indazolyl, Quinolinyl, Isochino linyl, naphthyridinyl, quinazolinyl. Pyridyl, pyrimidinyl, pyrazinyl are preferred and quinolinyl.

Halogen represents fluorine, chlorine, bromine or iodine, with fluorine and chlorine being preferred unless otherwise stated.

Preferred in the context of the present inventions are compounds of the general formula (I) in which
R ^{2 is} selected from the group

In the context of the present inventions, preference is also given to compounds of the general formula (I) in which
AC ₁ -C ₃ alkylidene.

In the context of the present inventions, preference is also given to compounds of the general formula (I) in which
R ^{1 is} cyclohexyl, pyridyl, thienyl, morpholinyl, phenyl or 2-pyrrolidinoyl.

The active substance can act systemically and / or locally. For this purpose he can be applied in a suitable manner, such as. B. oral, parenteral, pulmonary, nasal, sublin gual, lingual, buccal, rectal, transdermal, conjunctival, otic or as an implant.

The active ingredient can be administered in suitable administration forms for these administration routes be administered.

Known active ingredients are quick and / or suitable for oral administration modified dispensing application forms, such as. B. tablets (non-coated as well as coated tablets, e.g. (Enteric coatings), capsules, coated tablets, Granules, pellets, powders, emulsions, suspensions and solutions.

Parenteral administration can be done by bypassing an absorption step happen (intravenously, intraarterially, intracardially, intraspinally or intralumbally) or  with absorption (intramuscular, subcutaneous, intracutaneous, percutaneous, or intraperitoneally). For parenteral administration are suitable as application shape u. a. Injection and infusion preparations in the form of solutions, suspensions sions, emulsions, lyophilisates and sterile powders.

For the other application routes, z. B. Inhalation Drugs (et al. Powder inhalers, nebulizers), nose drops / solutions, sprays; lingual, sublingual or buccal tablets or capsules, suppositories, ear and Eye preparations, vaginal capsules, aqueous suspensions (lotions, shakes mixtures), lipophilic suspensions, ointments, creams, milk, pastes, powder or Implants.

The active substances can be added to the applications listed in a manner known per se forms are transferred. This is done using inert non-toxic, pharmaceutically suitable excipients. These include a. Carriers (e.g. micro crystalline cellulose), solvents (e.g. liquid polyethylene glycols), emulsifiers (e.g. sodium dodecyl sulfate), dispersant (e.g. polyvinylpyrrolidone), synthetic table and natural biopolymers (e.g. albumin), stabilizers (e.g. antioxidant such as ascorbic acid), dyes (e.g. inorganic pigments such as iron oxides) or taste and / or smell.

In general, it has proven to be advantageous for parenteral administration Amounts from about 0.001 to 10 mg / kg, preferably about 0.01 to 1 mg / kg body weight to give effective results. With oral application the amount is about 0.01 to 500 mg / kg, preferably about 1 to 10 mg / kg body pergewicht.

Nevertheless, it may be necessary from the amounts mentioned deviate, depending on body weight, application route, indi vidual behavior towards the active ingredient, type of preparation and time or interval at which the application takes place.  

The present invention further relates to a process for the preparation of the compounds of the general formula (I) by compounds of the general formula (II)

with compounds of the general formula (III)

H ₂ NR ² (III)

be converted into compounds of the general formula (I).

The present invention therefore also relates to the compounds of the general formula (II)

wherein R ¹ is cycloalkyl, cycloalkenyl, C ₆ -C ₁₀ aryl, a 4- to 10-membered, optionally fused heterocycle with up to 4 heteroatoms selected from the group O, S, N, where R ^{1 can} be optionally substituted with 1 to 3 substituents selected from the group halogen, trifluoromethyl, trifluoromethoxy, nitro, amino, alkylamino, dialkylamino, acylamino, oxo.

Determination of solubility

The compounds are dissolved in DMSO, and then the solution around the Diluted factor 100, d. H. there is still 1% DMSO in the solution. Then shake on the thermomixer at 25 ° C for 24 h. Then a Ultracentrifugation and the concentration in the supernatant by means of HPLC determined.

A calibration line is recorded for evaluation.

Example 11 shows a solubility of 11 mg / l.

Determination of the minimum inhibitory concentrations (MIC) in microtiter plates

The MIC values were determined using the microdilution method in Iso-Sen sitest broth performed. After serial dilution of the substances, the Inoculated microtiter plates with the test germs. The germ concentration was approx. 5 × 104 germs / ml suspension. After inoculation, the plates were left for approximately Incubated for 20 h at 37 ° C. The lowest concentration was proto as the MIC value collided, in which the visible growth of the bacteria was completely inhibited.  

The following table shows data in which the effect of Examples 3, 7 and 11 against Gram-positive bacteria (Staph. Aureus 133) with Gram-negative Bacteria and Candida are compared as comparative experiments on selectivity.

Table 1

Determination of the minimum inhibitory concentrations (MIC) compared to one Bunch of germs

The MICs of different bacterial strains were determined using the agar dilution method. MH agar + 10% horse blood + IsoVitale X was used as the nutrient medium. The inhibitors were tested in a concentration range from 0.25 to 64 µg / ml. The preparation concentrations were poured into the agar plates in two dilution steps. The bacterial strains were grown overnight in BH broth (staphylococci, enterobacteria) or BH broth + 10% bovine serum (streptococci, enterococci). The overnight cultures were diluted 1: 1000 in buffered, physiological saline. The agar plates were inoculated with these suspensions. This was done with the help of a multiple vaccination device (Denley Inoculator). The plates were incubated at 37 ° C under 8% CO ₂ . It was read after 18-20 hours. The concentration at which no growth could be seen macroscopically was given as the MIC value.

Table 2 below shows the activity of a compound (Example 11) against a number of Gram-positive germs:

Table 2

Systemic infection with S.aureus 133

S.aureus 133 cells were grown overnight in BH broth. The night culture was diluted 1: 100 in fresh bra broth and turned up for 3 hours. The bacteria in the logarithmic growth phase were centrifuged and 2 × with buffered, physiological saline (303) washed. Then a cell suspension was carried out on the photometer (Dr. Lange LP 2W) with an absorbance of 50 units in 303. After a dilution step (1:15) this suspension was 1: 1 with a 10% mucin suspension  mixed. 0.25 ml / 20 g of mouse were administered ip from this infection solution. This corresponds to a cell count of approximately 1 × 10E6 germs / mouse. The ip therapy took place 30 minutes after infection. Female CFW1- Mice used. The survival of the animals was recorded over 6 days.

In a group treated with the compound of Example 11, three of survived five mice by day 6 post infection compared to only one mouse in the untreated group (infection control).  

General synthesis instructions and manufacturing examples

Representation of urea from ammonium hydrochlorides

A mixture of primary amine in water (about 0.5 to 2.0 times / l) is added Ice cooling conc. Hydrochloric acid entered until the reaction is slightly acidic. alternative can also use the hydrochloride of the primary amine directly and in water be dissolved (0.5 to 2.0 mol / l). At room temperature, potassium or sodium cyanate (1.0 to 1.25 eq.) and the solution is for 2.0 to 6.0 h heated to reflux. After cooling, it usually sets directly and or after narrowing Failure of solid one. The solid is obtained by filtration, it will washed with water and dried thoroughly in vacuo. Alternatively, you can after removing the water in vacuo and taking up the residue with ethanol be filtered off in the heat; from the residue of the filtrate can then pass through Chromatography on silica gel (typically with solvent mixtures such as Toluene / methanol or dichloromethane / methanol) receive the product in pure form become.

Example I

N- [3- (Benzyloxy) propyl] urea, yield 21.1 g (83% of theory), melting point: 81 ° C

Representation of 1-substituted 2,4,6 (1H, 3H, 5H) -pyrimidine trions starting from ureas

Method A, basic reaction with ethanolate and malonic acid diester

To a solution freshly prepared from sodium and absolute ethanol (1.0 to 1.8 mol / l) of sodium ethanolate (1.8 to 2.5 eq.) in ethanol are urine under inert gas Substance derivative (1.0 eq.) and diethyl malonate (1.0 to 1.2 eq) added. The Mixture is heated under reflux for about 4 to 8 h and after cooling with salt acid to about pH = 1. All volatile components are removed in a vacuum removed. The residue is taken up in dichloromethane and washed with water rule. The organic phase is dried and evaporated to dryness in vacuo. If necessary, the product can be purified by chromatography on silica gel (eluent: mixtures of dichloromethane and methanol with the addition of vinegar acid).

Method B, acidic reaction with acetic anhydride and malonic acid (J. Hetero cyclic Chem., 1985, 22, 873)

To a solution of the corresponding urea derivative (1.0 eq.) And malonic acid (1.3 to 1.8 eq) in glacial acetic acid (approx. 1 to 3 mol / l) is vinegar under inert gas at approx. 60 ° C acid anhydride (approx. 3.0 eq.) added. At the end of the addition, the temperature increased and the mixture stirred at about 90 ° C for 4-8 h. After cooling, the volatile constituents of the reaction mixture removed in vacuo. From the back can be obtained by recrystallization (if necessary repeated) from alcoholic Solvents (e.g. ethanol) the product can be isolated. Alternatively, the Pro product can be obtained from the evaporation residue by RP-HPLC.  

Example II

1- (2-thienylmethyl) -2,4,6 (1H, 3H, 5H) -pyrimidintrione, melting point: 179 ° C

Preparation of 3-substituted 6-chloro-2,4- (1H, 3H) -pyrimidinediones (6-Chlorura cile)

A mixture of 1-substituted 2,4,6 (1H, 3H, 5H) pyrimidine trione (1.0 eq.), Ben cyltriethylammonium chloride (approx. 2.0 eq.) and phosphorus oxychloride (approx. 8 to 12.0 eq.) is heated to 50-60 ° C under inert gas for 2 to 4 h. After cooling, the reak added to ice water and carefully with concentrated sodium hydroxide solution (alternatively with concentrated ammonia solution) set to neutral (pH paper). The mixture is extracted thoroughly with dichloromethane. The organic phases are combined, dried and concentrated in vacuo. If necessary either purification of the product by chromatography on silica gel (elu mixtures of dichloromethane and methanol or ethyl acetate and ethanol) or by crystallization from dichloromethane.

Alternatively, 3-substituted 6-chloro-2,4- (1H, 3HJ-pyrimidinediones according to Literature regulations also by reaction with phenylphosphorus oxychloride (J. Hetero cyclic Chem., 1985, 22, 873) or by phosphorus oxychloride in the presence of less Amounts of water (J. Am. Chem. Soc., 1980, 102, 5036; tetrahedron asymmetry, 1997, 8, 2319) from the corresponding 1-substituted 2,4,6 (1H, 3H, 5H) - Represent pyrimidine trions.  

Example III

6-Chloro-3- [2- (2-pyridinyl) ethyl] -2,4 (1H, 3H) -pyrimidinedione, melting point: 184 ° C

Preparation of 3-substituted phenylamino-uracilene starting from 3-substituted 6-chloro-2,4- (1H, 3H) -pyrimidinediones

A mixture of 3-substituted 6-chloro-2,4- (1H, 3H) -pyrimidinedione (1.0 eq.) and aniline derivative (1.5 to 2.5 eq), optionally with the addition of a tertiary Amine base (approx. 2.0 eq.), In a high-boiling solvent (e.g. 1,4-dioxane, Diglyme or 1-methylpyrrolidinone) stirred for 1 h to 36 h at 100 to 150 ° C) before after aqueous work-up is concentrated in vacuo. The raw product can by chromatography on silica gel (eluents: mixtures of dichloromethane and Methanol) or by preparative RP-HPLC.

example 1 6 - [(2,2-difluoro-1,3-benzodioxol-5-yl) amino] -3- (3-methoxypropyl) -2,4 (1H, 3H) - pyrimidinedione, melting point: 235 ° C

The following examples were prepared according to the synthesis of Example I:

Preparation Examples

Claims (9)

1. Compounds of the general formula (I)
wherein
R ¹ is cycloalkyl, cycloalkenyl, C ₆ -C ₁₀ aryl, a 4- to 10-smooth, optionally fused heterocycle with up to 4 hetero atoms selected from the group O, S, N, where R ^{1 may} be substituted with 1 to 3 substituents selected from the group halogen, trifluoromethyl, trifluoromethoxy, nitro, amino, alkylamino, dialkylamino, acylamino, oxo,
R ^{2 is} a substituent of the following formula
wherein
R ^2-1 and R ^{2-2 are} independently selected from the group alkyl or halogen, or
R ^2-1 and R ^2-2 together form a 5- to 6-membered ring which can optionally have up to 2 oxygen atoms and which can optionally be substituted by up to 3 halogen,
A a group
wherein
n is a number from 0 to 5,
m is a number from 0 to 5,
L is -O-, -S-, -S (O) -, -S (O) ₂ -, -C (H) OH-, -C (O) -, -C = NR ¹⁰ - or a bond .
where R ¹⁰ is hydrogen, alkyl,
with the proviso that A is not CH ₂ when R ^{1 is} phenyl and that L is not equal to -C (O) - or -S (O) ₂ - when R ^{1 is} a ring bonded via a ring nitrogen and n is 0,
and their pharmaceutically acceptable salts, solvates and hydrates. 2. Compounds of general formula (I) according to claim 1, wherein
R ^{2 is} selected from the group
3. Compounds of general formula (I) according to claim 1 or 2, wherein
AC ₁ -C ₃ alkylidene. 4. Compounds of general formula (I) according to any one of claims 1 to 3, wherein
R ^{1 is} cyclohexyl, pyridyl, thienyl, morpholinyl, phenyl or 2-pyrrole dinoyl. 5. A process for the preparation of the compounds of the general formula (I) according to any one of claims 1 to 4, characterized in that connec tions of the general formula (II)
with compounds of the general formula (III)
H ₂ NR ² (III)
be converted into compounds of the general formula (I). 6. Compounds of general formula (I) according to any one of claims 1 to 4 to fight diseases.   7. Medicament containing compounds of the general formula (I) according to any of claims 1 to 4 and excipients. 8. Use of compounds of general formula (I) according to any of claims 1 to 4 for the manufacture of a medicament for treatment of bacterial diseases. 9. Compounds of the general formula (I)
wherein
R ¹ is cycloalkyl, cycloalkenyl, C ₆ -C ₁₀ aryl, a 4- to 10-membered, optionally fused heterocycle with up to 4 heteroatoms selected from the group O, S, N, where R ^{1 may} optionally be substituted by 1 up to 3 substituents selected from the group halogen, trifluoromethyl, trifluoromethoxy, nitro, amino, alkylamino, dialkylamino, acylamino, oxo.