DE102007037579A1 - New benzimidazol-2-yl-alkylamines and their use as microbicidal agents - Google Patents

Abstract

The present invention relates to novel benzimidazol-2-yl-alkylamines which have antimicrobial properties, to processes for the preparation of these compounds and to their use as microbicidal active substances in pharmaceutical preparations or crop protection agents. Although numerous microbicidal agents are known, due to the adaptability of the microorganisms and the associated formation of resistances there is a constant need for new, highly effective, but also physiologically compatible substances with as novel as possible mechanisms of action and broader spectrum of activity. The benzimidazole derivatives described have a good antimicrobial action, in particular against species of the genus Candida. Toxicity tests showed good compatibility of the new active ingredients. To prepare the compounds, two different methods are described. The compounds are used for the systemic and local treatment of humans, animals and plants infested with harmful microorganisms, in particular fungi, bacteria, viruses, algae, protozoa and other parasites. $ F1

Description

The The present invention relates to novel benzimidazol-2-yl-alkylamines, which have antimicrobial properties, process for the preparation of these compounds, for formulation as pharmaceutical agents and plant protection products and their use as microbicides Active substances or pesticides.

The The invention particularly relates to novel benzimidazol-2-yl-alkylamines with alkylbenzyl substituents in the 1-position and their use as active ingredients for the treatment of humans, animals and plants, those with harmful microorganisms, especially fungi, Bacteria, viruses, algae, protozoa and other parasites are.

State of the art

in the Over the years, many microbicidal agents have been used in agriculture, technology, human and veterinary medicine developed. In addition to inorganic and organometallic agents exists a very heterogeneous composite, difficult to understand Group of organic agents, of different substance classes belong to different impact areas.

The Antimicrobial effect of benzimidazole derivatives is already long known.

In pharmacy they are used as virucides, anthelmintics and antimycotics among others. Numerous pharmaceutically important benzimidazole derivatives are in the collections of Negwer / Scharnow as well as Elks / Ganellin listed.

in the technical field and in crop protection are benzimidazole derivatives used as pesticides, especially as systemic fungicides. An overview of the German-speaking

Perkow / Ploss published approved microbicidal agents that also contain a variety of benzimidazole derivatives for plant protection ,

Also The patent literature contains numerous benzimidazole derivatives for a wide variety of applications and medical indications. These are usually very specific substituted structures, which interfere with the enzyme metabolism of microorganisms. A especially important here are the benzimidazol-2-yl (alkyl) amides and benzimidazol-2-yl (alkyl) carbamates.

For example, SIRTRIS Pharmaceuticals has a group of benzimidazol-2-yl (alkyl) amides as sirtuin modulators for use in numerous medical indications such as diabetes, neurodegenerative diseases, cardiovascular diseases, blood clotting disorders, inflammation, cancer, as well in obesity, stress and aging signs revealed ( WO 2006/094209 . WO 2006/094235 ). In WO 2005/037272 the use of benzimidazol-2-yl (alkyl) amides to combat bacterial and parasitic infections is described. Benzimidazol-2-yl (alkyl) amide-structure microbicidal compounds, which are mainly used in agriculture, are in WO 2006/123145 . WO2007 / 020936 . US 3,920,680 and US 3,920,682 disclosed.

patents to benzimidazol-2-yl (alkyl) amines, very few have been published, wherein in most of the compounds we have become known, the amino group contains a secondary or tertiary nitrogen atom.

Secondary benzimidazol-2-yl-alkylamines as pharmaceutical agents for the treatment of cancer are, inter alia, in WO 2006/060737 and US 2006/0084687 released.

In WO2006 / 110683 . WO2005 / 028447 . WO2007 / 030080 and WO2002 / 32422 have been described primary benzimidazol-2-yl-alkylamines having defined substituents for use as enzyme inhibitors and corresponding pharmaceutical preparations. As a starting material for agricultural microbicides were in JP 08325235 benzothiazol-2-yl primary alkylamines and benzooxazol-2-yl-alkylamines.

The synthesis of primary benzimidazol-2-yl-alkylamines applies, especially in compounds with complex Substituents, generally difficult ( WO 2001/21634 ).

task

By Microbial diseases can be local or generalized occurrence. They occur, inter alia, disorders of the immune system, for example, in antibiotic or cytostatic therapies, in administration of steroids or hormones, according to Bestrah boys, at parenteral Nutrition, malignant diseases, endocrinopathies or immunodeficiencies. For systemic diseases, certain are preferred Organs affected. Skin disorders are often determined by certain Fungi, especially dermatophytes and yeasts. While Dermatophytes almost exclusively skin, hair and nails Mycoses also can be caused by yeasts on mucous membranes and internal organs.

plants are especially in unfavorable growing conditions like Climate, soils, nutrient deficiency / excess or monoculture susceptible to microbial attack.

Of the Mechanism of action of most microbicidal substances engages in the metabolism of microorganisms. Due to the high degree of adaptability Many microbes are a significant resistance. Even though Numerous antimicrobial agents are known, it follows a constant need for new, highly effective, as well Physiologically compatible substances with as possible novel mechanisms of action and broader spectrum of action.

specially We are looking for active ingredients that are resistant to such microorganisms who are already resistant to many known have developed microbicidal agents. In particular, fungal infections a problem in clinical and intensive care settings, which results in a constant need for new, well-tolerated Antifungals results.

Of the The present invention is therefore based on the object, new pharmaceutical active microbicides, in particular fungicidally active compounds, for use as anti-infective agents in humans and animals as well as To provide pesticides in crop protection, the remedy the mentioned disadvantages of conventional means.

It has now surprisingly been found that different primary benzimidazol-2-yl-alkylamine is an outstanding have antimicrobial activity, especially against Organisms that cause mycoses in humans, animals and plants can.

specially towards different fungi of the genus Candida, from some species are a problem especially for having antibiotics treated HIV-positive patients could be a very good fungicidal action of the substances of this new class of substances become.

sowie deren Verwendung als mikrobizide Wirkstoffe gegen Pilze, Bakterien, Viren, Algen, Protozoen und andere Parasiten.The present invention therefore relates to compounds of the formula

and their use as microbicidal agents against fungi, bacteria, viruses, algae, protozoa and other parasites.

Where R1 is a substituent from the following group:
branched or unbranched alkyl groups, in particular substituted or unsubstituted alkyl groups, substituted or unsubstituted alkylaryl groups, substituted or unsubstituted alkylheteroaryl groups, substituted or unsubstituted heteroaryl groups and side chains of natural and non-natural D and L amino acids may occur as side chains.

R2 represents a substituent from the following group:
H, alkyl, alkenyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, alkoxyalkyl, hydroxyalkyl, alkylmercapto, alky lamid, alkylcarboxy, alkylthioalkyl, N-substituted or N-unsubstituted alkylamine, haloalkyl, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, substituted or unsubstituted alkylaryl groups and substituted or unsubstituted alkylheteroaryl groups.

R3 and R4 in positions 4, 5, 6 and 7 are the same or different and are selected from the following substituent group:
H, halogen, trifluoromethyl, trifluoromethoxy, alkyl, alkoxy, thioalkyl, alkylthioalkyl.

A preferred embodiment relates to inventive Benzimidazol-2-yl-alkylamines with alkylbenzyl substituents in the 1-position according to the following formulas II to IX.

The antimicrobial activity of the compounds according to the invention and their physiological compatibility were demonstrated in an antifungal AS (activity-selectivity) assay. For this purpose, the half-maximal antifungal activity (IC ₅₀ ) of the test compounds and a reference compound on cell cultures infected with Candida albicans was determined.

For some particularly preferred variants of the compounds according to the invention, a ver comparable antimycotic activity as found for the highly effective amphotericin B commonly used as a reference standard.

investigations for the effectiveness of the compounds of the invention in three-dimensional cell culture models confirmed the pronounced antimicrobial effect. The compounds showed also good diffusion properties in the penetration of collagen and cell layers, so that an application of the invention Compounds as active ingredients for systemic and also topical Medicinal and plant protection products is possible. Negative influences on the cell cultures used were not observed.

In gene-based in vitro studies on the mode of action of the invention Connections were next to the already known attack points in the ergosterol biosynthesis also influences on the cell wall biogenesis detected by Candida albicans. This, for similar Compounds previously unobserved mechanism of action opens up new applications for the invention Compounds as active ingredients.

toxicity tests gave a good compatibility of the invention Links. It was in the relevant concentration range no observed toxic effect on the investigated cell lines.

The Spectrum of activity of the compounds of the invention includes fungi, bacteria, viruses, algae, protozoa and other parasites, especially yeasts, dermatophytes, molds, Pityrosporum ovale and biphasic mushrooms. The compounds of the invention can therefore be used successfully as active ingredients for treatment numerous local and systemic infections in humans, animals and plants are used. Particularly effective are the inventive Compounds for the treatment of dermatomycoses caused by Trichophyton rubrum, Trichophyton mentagrophytes and other trichophyton species, Microsporum canis, Epidermophyton floccosum and Scopulariopsis brevicaulis, as well as for the treatment of candidoses, Candida tropicalis, Candida albicans, Candida glabrata, Candida parapsilosis and other Candida types are caused.

typical Medical indications of microbial infections are, for example Disorders of the immune system, HIV infections (AIDS), diseases skin, hair and nails, mucous membrane infections, diseases of the respiratory and pharyngeal spaces such as pneumonia, bronchitis and Pharyngitis as well as inflammatory diseases of the inner Organs such as otitis, pyelonephritis, cystitis, endocarditis and arthritis.

at Plants are the compounds of the invention especially as pesticides for controlling mildew, mold fungi and other parasitic mushrooms.

One Another object of the invention is the preparation of the invention Benzimidazol-2-yl-alkylamines according to formula I, which according to the invention as microbicidal agents can be used.

One possible way is the synthesis of the invention Compounds on a solid phase starting from a nitroaniline (analogous to Scheme 1 in Example A). This is the output connection on a polymeric support with activated carbonic ester group immobilized and the resulting carbamate nitrogen in the basic Medium alkylated. The nitro group is then reduced and the resulting amino group with an aminocarboxylic acid derivative acylated. After acidic cleavage of the polymeric carrier resin the cyclization to benzimidazole in acetic acid solution. In a particularly preferred variant, the amino acid previously provided with an amino-protecting group after cyclization is split off.

at Another production variant is the synthesis of the invention Compounds carried out in solution (analogous to Scheme 2 in Example B). For this purpose, a Fluornitrobenzol with a Reacted amine. The nitro group is reduced and the product with an acylated Aminocarbonsäurederivat. This intermediate is then in acetic acid solution to benzimidazole cyclized. In a particularly preferred variant, the amino acid previously provided with an amino-protecting group after cyclization is split off.

Both Manufacturing methods allow both the synthesis of racemic as well as enantiomerically pure compounds.

The Starting materials for the preparation of the invention Connections are known and can be obtained from specialist dealers or manufactured according to known regulations.

One Another object of the invention is the use of the invention Benzimidazol-2-yl-alkylamines according to formula I for the preparation of pharmaceutical Agent for the treatment of human diseases and animals related to microbes. These anti-infectives, containing at least one inventive Active substance according to formula I, furthermore contain at least one or no non-toxic, inert, pharmaceutically acceptable carrier and at least one or no non-toxic, inert, pharmaceutically suitable auxiliary and / or additive.

pharmaceutical suitable materials are those in the field of pharmacy and food technology and in adjacent areas known to be usable Substances, in particular those in relevant pharmacopoeias listed, whose properties of a physiological application do not oppose.

The Pharmaceutical compositions of the invention containing at least one active ingredient of formula I, for example as tablets, dragees, capsules, pills, granules, suppositories, Injectables, Solutions, Suspensions and Emulsions, Pastes, Ointments, gels, creams, lotions, powders or sprays.

The Preparation of the pharmaceutical according to the invention Preparations are carried out in the usual way according to the person skilled in the art known methods.

The The invention also relates to methods for the treatment of diseases of humans and animals related to microbes. For this one gives to a person or an animal, the or the Such a treatment requires an antimicrobial Amount of a preparation according to the invention containing at least one compound according to the invention Formula I. The active ingredient or the invention Pharmaceutical preparation may be local, oral, parenteral, intraperitoneal and / or rectally, preferably orally or locally.

One Another object of the invention is the use of the invention Benzimidazol-2-yl-alkylamines according to formula I for the preparation of crop protection agents for the treatment of diseases with microbes in Related. These pesticides containing at least one active ingredient according to formula I according to the invention furthermore at least one or no non-toxic, inert, for the agriculture suitable carrier as well as at least one or non-toxic, inert, for agriculture suitable auxiliary and / or additive.

suitable Materials are those in the field of agriculture and food technology and in adjacent areas known to be usable Substances, in particular those in the relevant directories listed compounds, whose properties have a Application in agriculture.

The Pesticide according to the invention, containing at least one active ingredient of formula I, for example as granules, solutions, suspensions, powders or sprays be formulated.

The Production of the crop protection agents according to the invention is carried out in the usual manner by those skilled in the known methods.

The The invention also relates to methods for the treatment of diseases of plants or parts of plants associated with microbes stand. For this purpose, the plants or agricultural products, who require such treatment, with an antimicrobial acting amount of a preparation according to the invention, containing at least one compound of the invention according to formula I, pollinated, sprayed or the surrounding Soil enriched with it. The active ingredient or the invention Plant protection products can be applied locally or systemically preferably systemically by spraying the plants and Plant parts.

embodiments

Example A - Synthesis of a benzimidazol-2-yl-alkylamine with chlorinated alkylbenzyl substituent in the 1-position to solid phase

• 
  Scheme 1: Synthesis of a compound of the invention on a solid phase.

Loading of p-nitrophenyl carbonate Wang resin (X) with 2-nitroaniline (XI)

p-Nitrophenyl carbonate Wang resin (2 g, 3 mmol, loading 1.5 mmol / g) and 2.5 equiv. 2-Nitroaniline (7.5 mmol, 1.036 g) was weighed together into a filter frit syringe. A suspension of sodium hydride (3 equiv, 9 mmol, 216 mg of the 60% suspension) in 20 ml of DMF (dry over molecular sieve) added and the batch shaken for 18 h at room temperature.

The reaction solution was filtered off and the resin was washed with DMF until the washing solution was almost colorless. To the resin were added DMF (30 ml) and 25% NH ₃ in H ₂ O (3 ml) and shaken for 1 h at room temperature.

After filtering off the reaction solution, the loaded resin was washed (5 × DMF, 3 × MeOH, THF, DCM, Et ₂ O each time) and dried in vacuo.

Alkylation of o-nitroaniline carbamate Wang resin (XII)

In a syringe with filter frit became the o-nitroaniline carbamate Wang resin (3 mmol) a solution of LiOtBu (12 equiv; 2.88 g) in 20 ml of a mixture of anhydrous THF and anhydrous DMSO (1: 1). 3-Chlorobenzyl bromide (5 equiv., 205.48 g / mol, 1.964 ml) was added and shaken for 18 h at room temperature.

After filtering off the reaction solution, the resin was washed three times each with DMF, MeOH, THF, DCM and Et ₂ O and dried in vacuo.

Reduction of the nitro group of (XIII)

By 20 ml of a 2 M solution of SnCl ₂ x 2H ₂ O in DMF (13.3 equiv .; 40 mmol; 4.5 g / ml) was bubbled argon for several minutes. This solution was added to the resin and the reaction mixture shaken for 18 h at room temperature. The reaction solution was filtered off and the resin was washed (5x DMF, 3x MeOH, THF, DCM, Et ₂ O).

Acylation of (XIV) with Fmoc-L-valine

In a syringe with filter frit became the resin (3 mmol), Fmoc-L-valine (2.5 equiv., 7.5 mmol, 2.55 g) and PyBroP (2.5 equiv., 3.50 g). After addition of 25 ml NMP (dry over molecular sieve) and DIPEA (5 equiv .; 15 mmol; 1.94 g; 2.57 ml), the reaction was allowed to proceed at room temperature for 18 h shaken.

The reaction solution was filtered off and the resin was washed (5x DMF, 3x MeOH, THF, DCM, Et ₂ O).

Cleavage from the resin and cyclization to Benzimidazole (XVII)

The Cleavage from the polymeric support was carried out with 25% TFA in DCM.

To Shaking at room temperature for 1 h became the reaction mixture filtered, the filtrate caught on and the resin twice more 25% TFA washed in DCM. The washing solutions were combined with the cleavage solution.

To evaporation of the solvents and TFA became the crude product mixed with glacial acetic acid and the cyclization over 16 h at 80 ° C performed.

The Acetic acid was evaporated on a rotary evaporator in vacuo. The residue was prepared from tert-butyl alcohol / water (4: 1). lyophilized.

Cleavage of the Fmoc protecting group to (XVIII)

The Fmoc-protected crude product was mixed with a solution 25% piperidine in DCM and 1 h at room temperature touched.

The Reaction solution was on a rotary evaporator in vacuo evaporated to dryness. Purification of the product (XVIII) was carried out by silica gel chromatography.

Example B - Solution Synthesis a benzimidazol-2-yl-alkylamine with alkylbenzyl substituents in 1-position in solution

• 
  Scheme 2: Synthesis of a compound of the invention in solution.

Reaction of fluoronitrobenzene with 3-methylbenzylamine

To a solution of 1-fluoro-2-nitrobenzene (XIX) (20 mmol; 2.11 ml) in 50 ml of anhydrous THF was added 3-methylbenzylamine (XX). (20 mmol, 2.50 ml). The reaction solution was washed with Triethylamine (40 mmol, 5.57 ml) was added and refluxed for 12 h touched. The cooled reaction solution was then concentrated in vacuo and the crude product (HPLC, 214 nm,> 80%) continued without purification.

Reduction of the nitro group of (XXI)

N- (3-methylbenzyl) -2-nitroaniline (XXI) was taken up in 100 ml of ethanol / water (1: 1, v / v). After adding of sodium dithionite (80 mmol, 13.9 g) became the reaction solution Stirred at 70 ° C for 6 h.

The Reaction solution was extracted three times with 100 ml DCM each time. The combined organic phases were saturated with 100 ml NaCl solution, dried over sodium sulfate and freed from solvents in vacuo.

• Ausbeute: 3,10 g (73%, über zwei Stufen), C₁₄H₁₆N₂, M = 212,3 g/mol
• Reinheit (LCMS, 214 nm): 99%

The crude product of N- (3-methylbenzyl) -o-phenylenediamine (XXII) was purified by column chromatography on silica gel (eluent: DCM).

• Yield: 3.10 g (73%, over two steps), C ₁₄ H ₁₆ N ₂ , M = 212.3 g / mol
• Purity (LCMS, 214 nm): 99%

Acylation of phenylenediamine (XXII)

To a solution of N- (3-methylbenzyl) -o-phenylenediamine (XXII) (1.5 mmol, 318 mg) in 20 ml of anhydrous DCM became N-α-Boc-L-valine (1.8 mmol, 391 mg), N- (3-dimethylaminopropyl) -N-ethyl-carbodiimide hydrochloride (3 mmol, 573 mg) and a catalytic amount of 4-dimethylaminopyridine (10 mg) and stirred for 12 h at RT.

To Termination of the reaction became the reaction solution three times extracted with 50 ml of water, the organic phase over Sodium sulfate dried and in vacuo of solvents freed.

• Ausbeute: 570 mg (92,7%), C₁₄H₁₆N₂, M = 411,6 g/mol
• Reinheit (LCMS, 214 nm): 98%

N ¹ - {2 - [(3-Methylbenzyl) amino] phenyl} - (N-Boc) -valinamide (XXIII) was obtained as a tan solid.

• Yield: 570 mg (92.7%), C ₁₄ H ₁₆ N ₂ , M = 411.6 g / mol
• Purity (LCMS, 214 nm): 98%

Cyclization to benzimidazole (XXIV) and Cleavage of the protecting group

N ¹ - {2 - [(3-Methylbenzyl) amino] phenyl} - (N-Boc) -valinamide (XXIII) (1.36 mmol, 560 mg) was taken up in 30 mL of glacial acetic acid and stirred at 80 ° C for 8 h ,

After completion of the reaction, the reaction solution was freed from solvents in vacuo. (1S) gave 2-methyl-1- [1- (3-methylbenzyl) -1H-benzimidazol-2-yl] - (N-Boc) propyl amine (XXIV) (in the form of a brownish solid C ₂₄ H ₃₁ N ₃ O ₂ , M = 393.53 g / mol).

The thus obtained crude product was taken up in 15 ml TFA / water (2: 1) and stirred for 3 h at room temperature.

The Reaction solution became cautious under ice-cooling neutralized with a 2 M NaOH solution (pH 7-8) and extracted three times with 100 ml DCM each time. The combined organic Phases were washed with saturated NaCl solution and Water (3 x 100 ml), washed over sodium sulfate dried and freed from solvents in vacuo.

• Rohausbeute: 379 mg (94,8%), C₁₉H₂₃N₃, M = 293,42 g/mol
• Reinheit des Rohproduktes (LCMS, 214 nm): 81%

(1S) -2-Methyl-1- [1- (3-methylbenzyl) -1H-benzimidazol-2-yl] propylamine (XXV) was obtained as a tan solid.

• Crude yield: 379 mg (94.8%), C ₁₉ H ₂₃ N ₃ , M = 293.42 g / mol
• Purity of the crude product (LCMS, 214 nm): 81%

The Purification of the crude product was carried out by crystallization from EtOH / water.

Example C - Detection of fungicidal Effect on an antifungal AS assay

The antifungal activity and tolerability the compounds of the invention were based on an antifungal activity-selectivity (AS) assay. The results were compared to the values for amphotericin B (a industry-standard comparison standard for fungicides Compounds).

For this purpose, 10,000 HeLa cells per well of a microtiter plate with 5-500 (optimum: 50) CFU (MOI = 0.00025-0.025, optimum: 0.005) of Candida albicans (SC 5314, sequenced strain) in a total volume of 200 μl Medium (eg RPMI 1640, 10% FCS, 1% glutamine) in the presence / absence of the various drugs to be tested and incubated for 3-5 days at 37 ° C and 5% CO ₂ . From the number and vitality of the surviving cells, which were determined with a fluorescent dye, the IC ₅₀ and CC ₅₀ values were determined.

The analysis of the viable cells in the assay was after Kleymann / Werling carried out as follows. After the respective incubation time in the AS assay, the wells of the microtiter plate were washed with phosphate-buffered saline (PBS, 200 μl) and then filled with 200 μl PBS (containing 10 μg / ml FDA). After 20-45 min incubation at room temperature, the fluorescence (relative fluorescence units, RFU) with a wavelength of 485 nm was excited and the emission measured at 538 nm. (These values are for fluorescein. Other fluorophores are considered to have other appropriate wel lenlängen.)

ist ein Maß für die antimikrobielle Aktivität der getesteten Verbindungen. Tabelle 1: Relative antimykotische Aktivität diverser erfindungsgemäßer Verbindungen im Vergleich zur Referenzverbindung Amphothericin B; halbmaximale antimykotische Aktivität in Zellkultur gegenüber Candida albicans. Test-Verbindung Molekulargewicht [g/mol] IC₅₀ [μM] Referenz Amphotericin B 924,09 0,2–0,5 (IX) 313,80 0,25 (VI) 293,42 2 (VII) 313,83 2 (V) 327,86 4 (VIII) 299,81 5 (II) 311,41 30 The relative fluorescence intensities of the samples and an untreated control were compared. This quotient

is a measure of the antimicrobial activity of the tested compounds. Table 1: Relative antifungal activity of various compounds of the invention compared to the reference compound amphotericin B; Semi-maximal antifungal activity in cell culture against Candida albicans. Test compound Molecular weight [g / mol] IC ₅₀ [μM] Reference Amphotericin B 924.09 0.2-0.5 (IX) 313.80 0.25 (VI) 293.42 2 (VII) 313.83 2 (V) 327.86 4 (VIII) 299.81 5 (II) 311.41 30

Example D - Detection of fungicidal Effect in the cell model

The activity of the compound (IX) according to the invention against Candida albicans was investigated in three-dimensional cell culture models of a vaginal model which is composed of a collagen matrix and a cell layer growing thereon ( DE 10 06 2626 B4 ).

The test was carried out according to a publication by Dieterich et al. carried out. The substance was applied in such a way that it had to be diffused through the collagen matrix and the cell layer in order to come into contact with Candida albicans.

In 1 the cell cultures for the control without drug and in 2 the cell cultures for a sample spiked with the test compound (IX) are shown after 20 h of incubation time.

The Effectiveness of the substance could be confirmed. negative Influences on the vaginal model were in the relevant concentration range not to watch.

Example E - Examination the mode of action

The Mode of action of the compound according to the invention (IX) was tested in vitro by genome-wide DNA chips on Candida albicans examined.

The work was carried out as described by Sohn et al. described carried out. The induced genes were identified by an analysis of the transcription profile.

Out These studies concluded that ergosterol biosynthesis an essential target of the invention Represents connections. In addition to the ergosterol biosynthesis are unusually many Cell wall genes induced, what so far in azoles usually was observed only limited.

• Sc5314, YPD, 30°C, 3 h Induktion mit/ohne Verbindung (IX), (IC₅₀)
• Arrays: Fraunhofer IGB
• Datenanalyse: M-Chips

This means that the investigated compound (IX) according to the invention also has effects on the cell wall biogenesis, in particular on the β-1,3-; β-1,6-glucan biosynthesis. This leaves on new, for ä Similar compounds close previously unknown mechanisms of action and application potentials. Table 2: Genes found to be induced in the presence of (IX) using genome-wide microarrays. ERG pathway cell wall membrane +5.87 ERG25 +6.54 PGA23 +7.33 RTA2, keto, caspo +4.88 ERG251 +5.92 PHR2 +3.64 RTA3, keto, caspo +4.30 ERG6 +4.60 PGA45 +3.17 FTH1 +4.29 ERG11 +3.18 CRH11, caspo +2.77 hypothetical, ciclo +3.41 ERG5 +2.87 PGA7 +2.86 PHO87 +2.88 ERG1 +2.55 PGA52, flu +2.36 FRP1 +2.52 ERG24 +2.21 PGA10, keto, ciclo +2.28 CDR1 +2.47 ERG27 +2.20 CSH1, flu +2.44 ERG10 +2.11 HYR1 +2.37 ERG26 +2.05 KRE1 +2.05 UPC2 +1.91 RBT5 +1.93 ERG7 +1.90 ERG3

• Sc5314, YPD, 30 ° C, 3 h induction with / without compound (IX), (IC ₅₀ )
• Arrays: Fraunhofer IGB
• Data analysis: M chips

Example F - Examination of the toxicity

The Toxicity of the compound of the invention (IX) was tested on 3 cell lines.

A gradient screening with the cell lines CHO-K1 ( Chinese hamster ovary cells [ATCC Cat.No. CCL-61] ), A549 ( Human lung carcinoma [ATCC Cat.No. CCL-185] ) and HeLa ( Human cervix carcinoma [ATCC Cat.No. CCL-2] ) carried out.

It no toxic effect was observed on the cell lines used.

The half-maximal toxicity (CC ₅₀ ) for the investigated compound (IX) is approximately 100-150 μM.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• WO 2006/094209 [0009]
• WO 2006/094235 [0009]
• WO 2005/037272 [0009]
• WO 2006/123145 [0009]
• WO 2007/020936 [0009]
• - US 3920680 [0009]
• US 3920682 [0009]
• WO 2006/060737 [0011]
• US 2006/0084687 [0011]
• WO 2006/110683 [0012]
• WO 2005/028447 [0012]
• - WO 2007/030080 [0012]
• WO 2002/32422 [0012]
• - JP 08325235 [0012]
• WO 2001/21634 [0013]
• - DE 10062626 B4 [0080]

Cited non-patent literature

• - Chinese hamster ovary cells [ATCC Cat.No. CCL-61] [0089]
• - Human lung carcinoma [ATCC Cat.No. CCL-185] [0089]
• - Human cervix carcinoma [ATCC Cat.No. CCL-2] [0089]

Claims (23)

Benzimidazol-2-yl-alkylamine compounds of the formula

where R 1 is branched or unbranched alkyl groups, with side chains of substituted or unsubstituted alkyl groups, substituted or unsubstituted alkylaryl groups, substituted or unsubstituted alkylheteroaryl groups, substituted or unsubstituted heteroaryl groups and side chains of natural and non-natural D and L amino acids, R 2 is H, alkyl , Alkenyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, alkoxyalkyl, hydroxyalkyl, alkylmercapto, alkylamide, alkylcarboxy, alkylthioalkyl, N-substituted or N-unsubstituted alkylamine, haloalkyl, substituted or unsubstituted aryl groups, substituted or unsubstituted heteroaryl groups, substituted or unsubstituted alkylaryl groups, and substituted or unsubstituted alkylheteroaryl groups and R3 and R4 in positions 4, 5, 6 and 7 may be the same or different and is H, halogen, trifluoromethyl, trifluoromethoxy, alkyl, alkoxy, thioalkyl, alky lthioalkyl stand. A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A compound according to claim 1 characterized by the formula

A process for the preparation of benzimidazol-2-yl-alkylamine compounds according to any one of claims 1-9, characterized that a nitroaniline is immobilized on a polymeric support, then alkylated or arylated, subsequently reducing the nitro group, with a D or L amino acid or an aminocarboxylic acid derivative acylated and the intermediate after cleavage from the polymeric support is cyclized to benzimidazole. A process for the preparation of the benzimidazol-2-yl-alkylamine compounds according to any one of Ansprü characterized in that a Fluornitrobenzol reacted in solution with an amine, the nitro group is reduced, acylated with a D- or L-amino acid or an aminocarboxylic acid derivative and the intermediate is cyclized to benzimidazole. Benzimidazol-2-yl-alkylamine compounds according to a of claims 1-9, characterized in that they have an antimicrobial effect against fungi, bacteria, Viruses, algae, protozoa and other parasites. Use of benzimidazol-2-yl-alkylamine compounds according to any one of claims 1-9 for controlling of microbes characterized in that they are called microbicides Use active substances for the treatment of humans, animals and plants can. Method of combating fungi, bacteria, Viruses, algae, protozoa and other parasites, characterized that is an antimicrobial agent containing at least one A benzimidazol-2-yl-alkylamine compound according to any one of claims 1-9, directly on the microorganisms and / or their Habitat acts. Antimicrobial agent containing at least one A benzimidazol-2-yl-alkylamine compound according to any one of claims 1-9, characterized in that there is at least one or no vehicle and at least one or none Auxiliary and / or additive contains. An antimicrobial agent according to claim 15, comprising at least one benzimidazol-2-yl-alkylamine compound according to a of claims 1-9, characterized in that as carrier materials, auxiliaries and additives non-toxic, inert and pharmaceutically suitable materials from the field pharmacy or food technology. Process for the preparation of antimicrobial agents according to one of claims 15 and 16, characterized that at least one benzimidazol-2-yl-alkylamine compound according to any one of claims 1-9 with at least one or none of the other ingredients mixed and / or in solution brings. Antimicrobial pesticide containing at least one benzimidazol-2-yl-alkylamine compound according to a of claims 1-9, characterized in that there is at least one or no vehicle and at least contains no or no auxiliary and / or additive. Antimicrobial pesticide according to claim 18, containing at least one benzimidazol-2-yl-alkylamine compound according to any one of claims 1-9, characterized as carrier materials, auxiliaries and additives non-toxic, inert and application-technically suitable materials from the field agriculture or food technology. Process for the preparation of antimicrobial pesticides according to one of claims 18 and 19, characterized that at least one benzimidazol-2-yl-alkylamine compound according to any one of claims 1-9 with at least one or none of the other ingredients mixed and / or in solution brings. Method for the systemic treatment of diseases, associated with microbes, characterized that you give a person or an animal, or one of those Treatment requires a microbicidally effective amount of an antimicrobial Composition according to one of Claims 15 and 16, comprising at least one benzimidazol-2-yl-alkylamine compound according to one of claims 1-9 administered. Method for the topical treatment of diseases, associated with microbes, characterized that in the case of a person or an animal, or one of such Treatment requires a microbicidally effective amount of an antimicrobial Composition according to one of Claims 15 and 16, comprising at least one benzimidazol-2-yl-alkylamine compound according to one of claims 1-9, directly on the infected Applying tissue. Process for the systemic and local treatment of diseases of the plants associated with microbes, characterized in that plants or parts of plants in need of such treatment are contained with a microbicidally effective amount of an antimicrobial agent according to one of claims 18 and 19 at least one benzimidazol-2-yl-alkylamine compound according to any one of Claims 1-9, pollinated, sprayed or enriched the surrounding soil with it.