CS241098B2 - Method of new bis-triazole derivatives production - Google Patents

Description

The present invention provides a process for the production of novel bis-triazole derivatives which have antifungal activity and are useful in the treatment of fungal infections in animals, including humans, and as agricultural fungicides.

. Accordingly, the present invention provides a process for the preparation of compounds of formula (I)

in which

R is phenyl optionally substituted with 1 to 3 substituents independently selected from fluoro, chloro, bromo, iodo and trifluoromethyl, or is 5-chloropyrid-2-yl and their pharmaceutically and agriculturally useful salts.

The invention further provides an antifungal composition for use in agriculture (including horticulture) comprising a compound of formula I or an agriculturally acceptable salt thereof, together with an agriculturally useful diluent or carrier.

The invention also relates to a method of treating fungal-infected plants or seeds, characterized in that the plant or seed, or the place where the plant grows, is treated with an effective amount of a compound of the general formula. or of an agriculturally useful salt thereof.

Optionally substituted phenyl in R is preferably phenyl substituted with 1 or 2 substituents independently selected from the group consisting of fluorine, chlorine, bromine and iodine, and trifluoromethyl.

Preferred individual groups within the meaning of R are:

4-fluorophenyl,

4-chlorophenyl,

4-bromophenyl,

4-iodophenyl,

4-trifluoromethylphenyl,

- 2-chlorophenyl,

2,4-dichlorophenyl,

2.4- DifluorophenylOiva,

2-chloro-4-fluorophenyl,

Z-fluoro-4-chlorophenyl,

2.5-difluorophenyl,

2,4,6-trifluorophenyl.

4-bromo-2,5-difluorophenyl a

5-chloropyrid-2-yl.

The most preferred R groups are:

2,4-difluorophenyl,

2,4-dichlorophenyl,

4-fluorophenyl,

4-chlorophenyl a

5-chloropyrid-2-yl.

The most preferred individual compound of the invention is that wherein R ¹ is methyl and R 4 is chlorophenyl.

In accordance with the invention, the compounds of formula (I) above are prepared by producing a compound of formula (II)

OH

X - C 1 H 2 - C - CH - X ¹

I 1

R CH 3 (II) wherein

X and X ¹ are leaving groups, preferably chlorine, bromine or iodine atoms and

R is as defined in formula (I), reacted either with 1,2,4-triazole, preferably in the presence of a base, for example potassium carbonate, or with a salt of 1,2,4-triazole with a base, preferably a salt of 1,2, Of 4-triazole with an alkali metal.

Radicals represented by X and X ¹ are preferably the same and preferably represent a bromine atom.

Typically, the compound of formula (II), 1,2,4-triazole and potassium carbonate are heated in a suitable organic solvent, for example dimethylformamide, to a temperature of about 40 to 120 ° C until completion of the reaction. The resulting product can be isolated and purified in a conventional manner.

The starting materials of formula (II) may be prepared by a conventional method, for example according to the following reaction scheme:

O

II

R. MgBr + Br — CH2 — C — CH — Br CH5

OH

-> Br. CH 2 -, C-CH-Br

I I

R CH3. (ΠΑ)

The resulting intermediate of formula (II) does not need to be isolated, but can be used directly for the next reaction.

Pharmaceutically useful addition salts of the compounds of formula I are salts with strong acids forming nontoxic addition salts such as hydrochloric acid, hydrobromic acid, sulfuric acid, oxalic acid and methanesulfonic acid.

These salts can be prepared by conventional methods, for example by mixing solutions containing roughly equimolar amounts of the free base and the desired acid. The resulting salt is then isolated by filtration if insoluble or by evaporation of the solvent.

The compounds of the invention have been found to be unexpectedly well effective against infections caused by clinically relevant Aspergillus fungi.

The compounds of the formula I and their salts are antifungal agents useful for combating fungal infections in animals, including humans. For example, the compounds may be used in human medicine to treat local fungal infections caused, inter alia, by Candida, Trichophyton, Microsporum or Epidermophyton, or to treat mucosal infections caused by Candida albicans (e.g. oral candidiasis and vaginal candidiasis).

These compounds can also be used to treat systemic fungal infections caused by, for example, Candida albicans, Cryptococcus neoformans, Aspergillus flavus, Aspergillus fumigatus, Coccidioides, Paracoccidioides, Histoplasma or Blastomyces.

The in vitro evaluation of the antifungal activity of the disclosed compounds can be carried out by determining the minimum inhibitory concentrations of the test compounds in a suitable environment without growth of the respective microorganism.

In practice, a series of agar plates, each containing the test compound at the appropriate concentration, are inoculated with a standard culture, such as Candida albicans, and all plates are then incubated at 37 ° C for 48 hours. All plates are then found to grow, respectively. in the absence of fungal growth and the appropriate value of the minimum inhibitory concentration is calculated.

other microorganisms useful for these tests include Cryptococcus neoformans, Aspergillus fumigatus, Trichophyton spp., Microsporum spp., Epidermophyton floccosum, Coccidioides immitis and Torulopsis glabrata.

In vivo evaluations of the efficacy of the compounds of the invention can be performed by injecting mice inoculated with Candida albicans or Aspergillus flavus by intraperltoneal or intravenous injection, or orally by administering a series of different doses of the test compound.

Efficacy is then calculated based on the number of surviving specimens of the test animals in the treated group after the death of the untreated mice. The dose at which the test compound provides 50% protection against the lethal effect of infection (PD 50) is determined.

For use in human medicine, the antifungal compounds of formula I and their salts can be administered alone, but are generally administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice.

For example, they may be administered orally in the form of tablets containing such carriers as starch or lactose, or in the form of capsules or ovules in which they are contained either alone or in admixture with carriers, or in the form of elixirs or suspensions containing aromatic additives or dyes.

The disclosed compounds may be administered by parenteral injection, for example, intravenously, intramuscularly or subcutaneously. For parenteral administration, the disclosed compounds are best used in the form of a sterile aqueous solution which may contain other ingredients, for example, salts or glucose to render the solution isotonic.

When administered orally or parenterally in human medicine, the daily doses of the antifungal compounds of the formula I and their salts will range from 0.1 to 10 mg per kilogram, the total dose being administered in divided doses.

Thus, tablets or capsules will contain from 5 mg to; 0.5 g of active ingredient and will be administered once or more daily, as appropriate in each case.

The appropriate dose, which will be most suitable for ¹ patient, and will vary depending on the patient's age, weight and response, will be determined by the attending physician in each case.

The above dosage range is merely an example of an average case, and of course there will be individual cases where higher or lower doses are required. These cases are also within the scope of the invention.

Alternatively, the antifungal compounds of formula (I) may be administered in the form of suppositories or pessaries, or administered topically, in the form of a lotion, solution, cream, ointment or powder.

For example, these compounds may be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycols or liquid paraffin, or may be incorporated at a concentration of between 1 and 10% into an ointment base consisting of white wax or white soft paraffin, optionally with stabilizers and preservatives.

The compounds of formula I and their salts are also effective against a number of phytopathogenic fungi, including, for example, various rusts, mildews and molds, and the compounds described herein can therefore be used to treat plants and seeds for treating or preventing such diseases.

The in vitro activity evaluation of the compounds of the invention against phytopathogenic fungi is carried out by determining their minimum inhibitory concentrations using the same procedure as above, except that plates are incubated at about 30 ° C for 48 ° C prior to assessing fungal growth. hours or more.

Micro-organisms used in these tests include Cochliobolus carbonum,

Pyricularia -oryzae, Glomerella cingulata, Penicillium digitatum, Botrytis cinerea, and Rhizoctonia solani. In this test, the following minimum inhibitory concentration (MIC) values in µg / ml (six days incubation at 26 ° C) were found for the compound of Example 1 below. Experimental mushroom BALL (µg / ml) product from Example 1 Glomerella cingulata 200.0 Cochliobolus (Helminthosporium) carbonum 50.0 Penicillium digitatum 6.2 Pyricularia oryzae 100.0

For agricultural and horticultural applications, the compounds of the invention and their agriculturally useful salts are preferably used in the form of a composition adapted to the mode of application and the intended use. Thus, the compounds may be applied in the form of dusts, granules, seed dressings, aqueous solutions, dispersions or emulsions, baths, sprays, aerosols or fumes.

The disclosed compounds can also be supplied in the form of a dispersible powder, granulate or grains, or concentrates to be diluted prior to application. Such compositions may also contain conventional carriers, diluents or excipients as known and used in agriculture and horticulture, and may be manufactured by conventional techniques.

The disclosed compositions may also contain other active ingredients, for example compounds exhibiting herbicidal or insecticidal activity, or other fungicides.

The compounds of the invention and compositions containing them can be applied directly to the leaves, stems, branches, seeds or roots of plants, or to soil or other growth media, and can be used not only to treat disease but also prophylactically to protect plants or seeds from fungal infestation. .

The invention is illustrated by the following examples for guidance but are not intended to limit the invention in any way.

Example l. _t "

Preparation of 1,3-bis (1H-1,2,4-triazol-1-yl) -2- (4-chlorophenyl) baptap-27-yl

Mg8r

OH

N, N-CH 2 -CH (CH, Hf

N X Ν '

Cl ml of 4-chlorophenylmagnesium bromide iv ether (prepared from 15.2 g of 4-chlorobromobenzene and: 2.8 g of magnesium shavings) was added via syringe to a solution of 9.2 g of 1,3-dibromobutane-2 over 30 minutes under nitrogen. -one (Org. Synthesis, 53, 123) in 50 mL dry ether cooled to -78 ° C.

After stirring at -78 ° C for 1 hour, a saturated aqueous ammonium chloride solution was added to the resulting mixture, and the mixture was allowed to warm to room temperature. The ether layer was separated, the aqueous layer was extracted three times with 2: 0 ml of ether each time, the combined ether extracts were washed with brine and dried over magnesium sulfate. The ether evaporation residue was added to a mixture of 8 g of 1,2,4-triazole, 20 g of potassium carbonate and 50 ml of dimethylformamide under a nitrogen atmosphere, and the mixture was heated at 70 ° C overnight.

After cooling, the reaction solution is filtered, the solid residue on the filter is washed with 50 ml of xylene and the combined filtrates are evaporated in vacuo. The last dimethylformamide residues were removed by double azaotropic distillation in 30 ml xylene each.

The crude residue was partitioned between 200 ml of methylene chloride and 100 ml of water, the methylene chloride extract was washed with water and dried over magnesium sulfate. The methylene chloride was evaporated in vacuo and the residue subjected to flash chromatography on 150 g of silica gel using methylene chloride containing 8% methanol by volume.

Fractions of 50 ml are collected. The appropriate fractions (i.e. fractions 29-39) were combined and evaporated to give 1.1 g of a mixture of diastereomeric pairs. The isomeric vapors were separated by flash chromatography on 100 g of silica gel, eluting with ethyl acetate / diethylamine / methanol (80: 20: 2 by volume). Fractions 13-16 (each containing 25 mL) were combined and evaporated to give 114 mg of isomeric pair 1 which melted at 112-113 ° C after crystallization from ethyl acetate-hexane. Mass Spectrum: Calcd for Cm H 18 ClN 6 O m / e 318 (M ⁺ ) found m / e 318 (M ⁺ ).

Evaporation of fractions 31-49 (each again containing 25 ml) followed by crystallization from ethyl acetate / hexene gave 248 mg of isomeric pair 2, m.p. 50-51 ° C.

Mass spectrum: calcd. For C 11 H 15 ClN 5 O m / e 318 (M ¹ ) found m / e 318 (M ⁺ ).

Isomeric Pair 1:

NMR (CDCl3):

1.25 (doublet, J = 7Ηζ, 3H, CH 3),

3.76 (doublet, J = 13 Hz, 1H, N - CH3), i

4.32 (doublet, J = 13 Hz, 1H, N-CH2), 4.92 [quartet, J = 7 Hz, 1H, N-CH (CH3)], 5.48 (singlet, —OH, exchangeable with D2O) 7.12 (multiplet, 4H, C6H4), [7.55 (singlet, 1H), 7.70 (singlet, 1H), 7.92 (singlet, 1H), 8.30 (singlet, 1H), protons of the triazole residue].

found:

% C, 52.5.5;% H, 4.5%; N. 26.1%.

The following overview sets forth the PD50 values (mg / kg; oral administration) found for some of the compounds of the invention in the above-described Candida albicans-infected mouse test.

Isomeric Couple 2:

The product from Example #

PD50 (mg / kg)

NMR (deuterochloroform, <S):

1.53 (doublet, J = 7Hz, 3H, CH 3),

4.56 (singlet, 2H, N - CH2),

4.72 (doublet, J = 7 Hz, 1H, CH),

CH3

5.52 (singlet, OH, exchangeable with D2O),

6.95 (multiplet, 4H, C6H4), (7.65 (singlet, 1H), 7.78 (singlet, 2, H).

7.88 (singlet, 1H), protons of the triazole residue).

Example 2

Analogous to the previous example, the appropriate starting materials were prepared and separated into two diastereomeric pairs of 1,3-bis (1H-1,2,4-triazol-1-yl) -2- (2,4-difluorophenyl) butan-2-ol. Isomeric pair 1 melts at 186-188 ° C and has the following microanalysis:

Analysis calculated for: CHmHzFďNďO:

52.5% C, 4.4% H, 26.2% N;

(diastereomeric pair 1) 0.1 (diastereomeric pair 2) 0.3 (diastereomeric pair 2) 0.5

To test for activity against systemic aspergillosis in mice, mice are infected with an Aspergillus flavus strain by intravenous injection into the tail vein. Untreated (control) mice normally die within 5-10 days after infection with Aspergillus flavus.

Each test compound is administered orally at a dose of 20 mg / kg to a group of infected mice, 1 hour and 4 hours after infection, and then twice daily for 4 consecutive days. An increase in the average survival time (PDP) of the treated mice compared to a control group of mice infected at the same time with the same strain is observed.

The compounds of formula I have been found to exhibit unexpected activity against an important strain of Aspergillus flavus. The following is a comparison of the results obtained with analogous compounds containing the grouping "CH 2" in the chain (see British Patent Application No. 2,078,719A) instead of the grouping.

R ¹ found:

H, 4.5; N, 26.5.

Isomeric pair 2 melts at 123 to 125 ° C and has the following microanalysis:

Calcd for CuHuF2NeO:

52.5% C, 4.4% H, 26.2% N;

N N

CI-13

OH _R 1 ri p

'N

(diastereomer 1 of Example 1)

Cl ·

H + 2.4

Claims (6)

SUBJECT A process for the preparation of novel bis-triazole derivatives of the general formula I in which R represents a phenyl group optionally substituted by one to three substituents independently selected from the group consisting of fluorine, chlorine, bromine, iodine and trifluoromethyl groups or represents a 5-chloropyrid-2-yl group, and pharmaceutically or agriculturally acceptable salts thereof, characterized by The method of claim 1, wherein the compound of formula II OH X — CH2 — C — CH — X ¹ I I R CH 3 (II) wherein X and X ¹ are each a leaving group and R is as defined in formula I, reacted with 1,2,4-triazole or a salt thereof with a base, and optionally converted to a pharmaceutically or agriculturally useful salt. 2. A method according to claim 1, characterized by using starting materials of formula II, wherein both X and X ¹ represent a bromine atom and R is as defined above. ' 3. A process according to any one of Claims 1 and 2 wherein the reaction is carried out using 1,2,4-triazole in the presence of potassium carbonate. 4. A process according to any one of Claims 1 and 2 wherein the reaction is carried out using an alkali metal salt of 1,2,4-triazole. 5. A process according to any one of Claims 1 to 4, wherein the corresponding starting materials are used to form compounds of formula I wherein R is 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-iodophenyl, -trifluoromethylphenyl, 2-chlorophenyl, 2,4-dichlorophenyl, 2,4-difluorophenyl, 2-chloro-4-fluorophenyl, 2-fluoro-4-chlorophenyl, 2,5-difluorophenyl, 2,4,6-trifluorophenyl 4 is first a conversion of erythromycin A (I) to a rid-2-yl group. 6. A process according to claim 5, wherein the corresponding starting materials are used to form compounds of formula I wherein R is 2,4-difluorophenyl, 2,4-dichlorophenyl, 4-fluorophenyl, 4-chlorophenyl or 5-chlorophenyl. a chloropyrid-2-yl group.