JPH0233691B2 - SHINKIBISUUTORIAZOORUJUDOTAIOYOBISONOSEIHOOYOBISOREKARANARUKOSHINKINZAI - Google Patents

Description

[Detailed description of the invention]

The present invention relates to antifungal agents comprising novel bis-triazole derivatives that have antifungal activity and are useful in the treatment of fungal infections in humans and other animals. The present invention provides a compound represented by the following formula and a pharmaceutically acceptable salt thereof. British Patent No. 2078719A issued on January 13, 1982 and
European Patent No. 0044605 issued on January 27, 1982 describes a compound represented by the following formula, its salt, a metal complex,
Ethers and esters are disclosed. [R ¹ is optionally substituted alkyl, cycloalkyl (e.g., cyclopentyl, cyclohexyl), aryl (e.g., phenyl), or araltyl (e.g., benzyl); Y ¹ and Y ² are CH- or =N- ] These compounds are said to be useful as fungicides and plant growth regulators. It is also stated to be active against fungal diseases in humans. In particular, Example 2 and Claim 7 of these applications contain 1,3-bis(1,2,4-triazole-1-
yl)-2-(2,4-dichlorophenyl)-propan-2-ol is disclosed. This compound is represented by the following formula. The 2,4-difluorophenyl compound of the present invention is not specifically disclosed in these patent specifications, and "2,4-difluorophenyl" is particularly named as a good example of "R ¹ ". It hasn't even been raised. Although this 2,4-dichlorophenyl compound is teratogenic, the 2,4-difluorophenyl compound of the present invention is quite unexpectedly not teratogenic. Furthermore, the corresponding 2-,3-,4-chlorophenyl,
4-bromophenyl is teratogenic. The above was confirmed in the following teratogenicity study. Teratogenicity research Female rats after conception [Cr1: COBS-CD (SD)
BR, Chales River Breeding Colony (France)
were randomly divided into groups of 5 animals each. 6~ after conception
Test compounds were administered via gastric intubation for 10 consecutive days on day 15 as a suspension in 0.1% aqueous methylcellulose. Rats were sacrificed 20 days after conception, and the number of dead fetuses was recorded along with the number and weight of live fetuses. All fetuses were examined for abnormalities in appearance, mouth, and internal organs. The test compound is represented by the formula: (R is 2,4-dichlorophenyl, 2-, 3-,
4-chlorophenyl, 4-bromophenyl or 2,4-difluorophenyl) All fetuses from animals treated with a compound in which R is 2,4-dichlorophenyl [20 mg/Kg (body weight)] It showed malformations (particularly ruptured palate). Examination of visceral and skeletal characteristics revealed that it was teratogenic at doses as low as 1 mg/Kg (e.g. microphthalmia), increased the frequency of ureteral and renal meninges, delayed ossification of some bones,
It also increased the incidence of the 14th rib pair. For compounds where R is 4-chlorophenyl, 20 mg/
Kg is extremely embryotoxic, while the compound in which R is 2-chlorophenyl exhibited dysmorphic appearance (cleft palate) at this dose. These compounds are listed in Table 1 of the above-mentioned British Patent No. 2078719A as "Compound 1," respectively.
Furthermore, compounds in which R is 3-chlorophenyl, 4-bromophenyl (specifically disclosed in the specification but not included in the claims) are also disclosed as 20 mg/Kg. The latter was also embryotoxic at this dose. When the compound of the invention (R = 2,4-difluorophenyl) was administered to pregnant rats under the same conditions,
Surprisingly, no external malformations occurred. The internal organs of the fetuses from these animals were examined, but no significant abnormalities were found in the internal organs or skeleton. The embryotoxicity (embryo-teratogenicity and/or embryotoxicity) of compounds in which R is other than 2,4-difluorophenyl represents one of the greatest drawbacks when their use in human mycoses is desired. The compounds of this invention are administered with pharmaceutically acceptable diluents and carriers, if necessary. Preferably, it is used in tablets, capsules, injections, and ointments. Compounds of formula () can be obtained in a number of different ways. In one method of the invention, the formula: The oxirane represented by is obtained by reacting with 1,2,4-triazole, preferably in the presence of a base. Preferred bases are potassium carbonate and NaH. One typical method involves reacting (generally within 8 hours) 1,2,4-triazole, oxirane (), and anhydrous potassium carbonate in a suitable solvent (e.g., dry dimethylformamide) preferably heated to 50-120°C. completion) to promote the reaction. The product can be isolated and purified using conventional methods. The starting triazole can also be used as an oxidized salt, such as the methanesulfonate salt, but this requires the use of an excess of base. Oxirane () is a conventional method, typically corresponding ketone (): is obtained by reacting trimethylsulfoxonium iodide with dimethyloxosulfonium methylide prepared from cetrimide/NaOH. This reaction is typically carried out as follows. The ketone (), trimethylsulfoxonium iodide, and cetrimide are vigorously stirred in a mixture of toluene and aqueous NaOH for 2-3 minutes at a temperature up to about 100°C. The product oxirane can then be isolated in conventional manner. Ketone () can be produced by a conventional method, for example, as follows. The compound of formula () can also be produced as follows. Intermediate oxirane () (can be isolated if desired)
is thought to be formed in this reaction. In one typical reaction, the compound () and 1,2,4-
The triazole in a suitable solvent (e.g. N,N-dimethylformamide or tetrahydrofuran)
Heat together at a temperature up to 120° C. for up to about 24 hours, preferably in the presence of a base (eg potassium carbonate). Generally an excess of triazole and base is used. The product ( ) can then be isolated and purified using conventional methods. In both of the above processes, the product ( ) is generally mixed with isomers in which one of the triazole rings is attached to the adjacent CH ₂ in the 4-position. However, this unwanted isomer can be removed by chromatography (eg on silica gel) or recrystallization. The starting material () is obtained in a conventional manner. for example, Preferred pharmaceutically acceptable salts are acid addition salts. Pharmaceutically acceptable acid addition salts of compounds of formula () are prepared from strong acids to form non-toxic acid addition salts containing pharmaceutically acceptable anions, such as hydrochloride, hydrobromide, sulfate, etc. It is something that is formed. The salt is prepared in a conventional manner, eg, by mixing a solution containing equimolar amounts of the free base and the desired acid, and collecting the desired salt by filtration (if insoluble) or by solvent evaporation. The compounds of formula () and their pharmaceutically acceptable salts are highly active antibacterial agents and are useful in the treatment of fungal infections in animals, including humans. For example, among others
Candida, Trichophyton, Microsporum,
Localized fungal infections in humans caused by Epidermophyton species, mucosal infections caused by Candida albicans (e.g.
Useful in treating thrush and vaginal candidiasis). For example, Candida, albicans, Cryptococcus,
Paracoccidioides, Histoplasma, Blastomyces
It can also be used to treat systemic fungal infections caused by. In-hydro evaluation of the antifungal activity of a compound () can measure the minimum inhibitory concentration (mic), which is the concentration of the test compound in a suitable medium at which no microbial growth occurs. In practice, for example, Candida
albicans, and then at 37°C.
Incubate for 48 hours. Next, check for the presence or absence of fungal growth and record the appropriate mic value. Other microorganisms used in such tests are Cryptococcus
neoformans, Aspergillus fumigatus,
Trichophyton spp，Microsporum spp，
Epidermophyton floccosum, Coccidioides
immitis, Torulopsis glabrata, etc. In vivo evaluation of compounds can be performed by administering a series of doses intravaginally, intravenously, or orally to mice inoculated with a Candida albicans strain. Untreated mice die within 48 hours and the compound dose that protects 50% from the lethal effects of infection is recorded. Compound () provides at least 50% protection at 0.5 mg/Kg (po. or iv). The antifungal activity of the compounds of the invention against (a) acute systemic candidiasis and (b) vaginal candidiasis was compared in vivo with the compound of the aforementioned GB 2078719A. (1) In testing the activity against (a) above, first, normal mice were inoculated with Candida, a species preserved by Pfizer.
The animals were infected with C. albicans by tail vein injection.
All untreated (control) mice died of infection within 48 hours. Appropriate doses of compounds were administered orally or intravenously to groups of infected mice at 1, 4 and 24 hours post-infection. From the results of the number of surviving mice after 48 hours at each dose of the compound,
Regression analysis was used to calculate the dose required to prevent half of the deaths in the treatment group. Next, regarding the activity against similar infections,
"Immunosuppressed" mice, ie, mice treated with cyclophosphamide to reduce neutrophils and thereby reduce their resistance to infection, were also tested. Furthermore, in order to reduce the variation from test to test,
Compounds were tested against the same infection of both normal and "immunosuppressed" mice simultaneously in parallel to provide a direct comparison. (2) In the test for activity against (b) above, mice maintained in estrus were intravaginally infected with Candida albicans, the same species as in (a), and immediately after infection, the test compound was administered to a group of infected mice. 20~0.625
mg/Kg was administered orally. The effect is % reduction of Candida albicans present in vaginal smear after 10 days.
Mycologically evaluated and measured. (3) The following compounds were used in the test.

【table】 (4) The test results are shown below: (a) Acute systemic candidiasis

【table】

【table】

【table】 (b) Vaginal candidiasis

[Table] Acute toxicity value (LD ₅₀ ) of antifungal compound of formula ()
was 1524 mg/Kg by oral administration in mice. For human use, the antifungal compound of formula (or a salt thereof) can be administered alone, but will generally be administered in admixture with a pharmaceutical carrier selected having regard to the route of administration and standard pharmaceutical practice. Ru. For example, it can be administered orally in the form of tablets containing excipients such as starch or lactose, capsules or oval preparations alone or in combination with excipients, and elixirs or suspensions containing flavorings and colorants. . For example, it can be injected parenterally intravenously, intramuscularly, or subcutaneously. For parenteral administration, it is best used in the form of a sterile aqueous solution which may contain other substances, such as salts and glucose in sufficient quantities to render the solution isotonic. For oral or parenteral administration to humans, the daily dose of the antifungal compound of formula (), whether oral or parenteral, is expected to be 0.1 to 5 mg/Kg (administered in divided doses).
Therefore, tablets and capsules of the compound may contain 1
It may contain 5 mg/0.5 g of the active compound, given once or more than once. In each case, the physician will determine the actual amount that is most appropriate for each patient, and it will vary depending on the age, weight, and responsiveness of each patient. The above doses are examples of the average case. There may, of course, be cases where greater or lesser amounts are beneficial, and these cases are also within the scope of the present invention. Alternatively, antifungal compounds of formula () can be administered in the form of suppositories or petals, and can also be used topically in the form of lotions, solutions, ointments, dusting powders. For example, it can be blended into an ointment made of an aqueous emulsion of polyethylene glycol or liquid paraffin, or it can be blended into an ointment made of a white wax or white soft paraffin base together with necessary stabilizers and preservatives at a concentration of 1 to 10%. The following examples are illustrative of the invention. Example 1 2-(2,4-difluorophenyl)-1,3
-Bis(1H-1,2,4-triazole-1
-il) propane-2-ol (i) 1-bromo-2,4-difluorobenzene (0.96g, 5mM) in diethyl ether (10ml)
The mixture was stirred at -78°C, and a hexane solution of n-butyllithium (1.55M, 3.23ml; 5mM) was added over 3 minutes. After the addition was complete, the mixture was stirred for an additional 10 minutes, and then a solution of 1,3-dichloroacetone (0.63 g, 5 mM) in diethyl ether (10 ml) was added dropwise over 3 minutes. After stirring for an additional 30 minutes at −78°C, acetic acid (0.33 g) in diethyl ether (5
ml) solution at 0°C, then water (10ml) was added. The organic layer was separated and the aqueous layer was washed once with diethyl ether. The combined ethereal extracts were dried (MgSO ₄ ) and evaporated to give a pale yellow oil which was dissolved in dimethylformamide (20ml). This DMF solution contained intermediate (IVB). (ii) Add 1,2,4-triazole (1.72g, 25mM) and anhydrous potassium carbonate (2.07mM) to the solution prepared in (i).
g, 15mM) and heated at 70°C for 18 hours.
Cooled and poured into water (100ml). Extracted twice with ethyl acetate. The organic extract was dried (MgSO ₄ ) and evaporated to a gum. This gum-like substance was subjected to chromatography [Silica (270-400
methylene chloride) to give the title compound as a white solid, 0.40 g (26% based on dichloroacetone), mp (after crystallization from ethyl acetate/hexane) 138-140°C. Analysis (%): Measured value: C, 51.33; H, 4.05; N, 27.08 Calculated value C ₁₃ H ₁₂ F ₆ N ₆ O:
C, 50.98; H, 3.95; N, 27.44 ¹ HNMR, IR, and mass spectral data were consistent with the required structure. The desired product is isolated by chromatography from impurities present in the reaction mixture.
[2-(2,4-difluorophenyl)-2-hydroxy-3-(4H-1,2,4-triazol-4-yl)-propyl]-1H-1,2,4
-Separated from triazole. Example 2 (A) 2-chloro-2',4'-difluoroacetophenone Chloroacetyl chloride (113g, 1.0M) 1,
The mixture was added dropwise to a stirred mixture (room temperature = 20°C) of 3-difluorobenzene (114g, 10M) and anhydrous aluminum chloride (146.6g, 1.1M). 50~55℃
The mixture was further stirred for 5 hours. Methylene chloride (48.5 ml) was slowly added while allowing the mixture to cool to room temperature. Separate the methylene chloride layer, wash with water (2 x 320 ml), and remove the solvent under reduced pressure to give a pale yellow solid (180 g).
I got it. A portion of this crude product (145 g) was crystallized from n-hexane (435 ml) to give the title compound (113
g, 73%), mp47-49℃ [Reference D. Ehlers, H.
Bercher and A. Grisk, J. Prakt, chem.,
315, 1169 (1973) value of 46.5℃] was obtained. IR
(KBR) and NMR (CDCl ₃ ) were consistent with the desired structure. (B) 2′,4′-difluoro-2-(1H-1,2,4
-triazol-1-yl)acetophenone
HCl 1,2,4- in ethyl acetate (186 ml)
A mixture of triazole (30.4 g, 0.44 M) and triethylamine (15.1 g, 0.15 M) was treated with 2-chloro-2',4'-difluoroacetophenone (38.1 g, 0.2 M) in ethyl acetate (80 ml). solution was added. The mixture was refluxed for 6 hours and then cooled to room temperature to remove insoluble matter. Pour the liquid into water (2 x 200ml)
The solvent was then distilled off under reduced pressure. The crude product was dissolved in ethyl acetate (150ml) and then 25% W/V HCl gas in isopropanol was added.
The mixture was granulated at 0° C. for 1 hour, then the solid was removed and dried to give the title compound (21.6 g, 40%),
mp167~170℃, obtained. IR (KBr), NMR
(DMSO) spectrum was consistent with the desired structure. This intermediate was characterized as the free base (prepared as follows). Bicarbonate of soda (16.8g, 0.2M) and 1,2,
A solution of 2-chloro 2',4'-difluoroacetophenone (38.1 g, 0.2 M) in toluene (45 ml) to a stirred slurry in refluxing toluene (180 ml) with 4-triazole (27.6 g, 0.4 M). added. The mixture was stirred at reflux for 3 hours and the water formed during the reaction was removed using a Dean-Stark trap.
The reaction mixture was cooled to room temperature and then diluted with water (180
ml) was added. The toluene layer was separated and the solvent was removed under reduced pressure. The resulting light brown solid was crystallized from 1:1 ethyl acetate:n-hexane (70ml) to give the title compound (3.9g), mp 103-105°C. IR
(KBr) and NMR (CDCl ₃ ) spectra were consistent with the desired structure. Analysis (%): Calculated value C ₁₀ H ₇ F ₂ N ₃ O:
C, 53.8; H, 3.16; N, 18.82. Measured value: C, 56.62; H, 3.15; N, 18.68. (C) 1-[2-(2,4-difluorophenyl)
-2,3-epoxypropyl]-1H-1,2,
4-triazole methanesulfonate 2′,4′-difluoro-2-(1H-1,2,4
-triazol-1-yl)acetophenone
HCl (59.6g, 0.23M), trimethylsulfoxonium iodide (50.6g, 0.23M), and cetrimide (2.1g) were mixed with toluene (370ml) at 60°.
The mixture was stirred for 3 hours in a mixture with a 20 w/v % NaOH aqueous solution. The toluene layer was separated and concentrated to 110ml, then diluted with ethyl acetate (150ml). A solution of methanesulfonic acid (16.6g, 0.172M) in ethyl acetate (20ml) was added. Ethyl acid acid (100 ml) was added and stirred at 0°C for 1 hour, and the title compound (43
g, 56%). 20 g of this crude product was dissolved in industrial methylated spirits (140 ml) and charcoal (2 g) was added. filtrate, concentrate the liquid to 100ml, and then
Stirred at ℃ for 1 hour. The title compound (7.8 g, 39%) mp 128-129°C was obtained by filtration. IR
(KBr) and NMR (DMSO) spectra were consistent with the desired structure. Analysis (%): Calculated value C ₁₂ H ₁₃ F ₂ N ₃ O ₄ S:
C, 43.2; H, 3.9; N, 12.6. Measured value: C, 42.83; H, 3.92; N, 12.96. (D) 2-(2,4-difluorophenyl)-1,
3-bis(1H-1,2,4-triazole-
1-yl)-propan-2-ol 1-[2-(2,4-difluorophenyl)
-2,3-epoxypropyl]-1H-1,2,
4-triazole methanesulfonate (6.7g,
0.02M), 1,2,4-triazole (2.8g,
0.04M), anhydrous potassium carbonate (9.1g, 0.066M)
in dimethylformamide (35 ml) at 90°C.
Stirred for 1/2 hour. After cooling to room temperature, cool with water (170
ml). It was extracted with chloroform (2 x 60 ml) and the extracts were combined and washed with water (2 x 100 ml). Dry the chloroform solution (MgSO ₄ ),
The solvent was distilled off under reduced pressure to obtain a crude product (5.3 g). This crude product was dissolved in isopropanol (50ml) and charcoal (0.5g) was added. The solution was concentrated to 25 ml. The precipitate was collected and dried to give the title compound (12.6 g, 44%); mp 139-140°C. The IR (KBr) and NMR (DMSO) spectra were consistent with the desired structure. Analysis (%) Calculated value C ₁₃ H ₁₂ F ₂ N ₆ O:
C, 51.0; H, 3.92; N, 27.5. Measured value: C, 50.85; H, 3.92; N, 27.74. Formulation example 1 Capsule Ingredients Amount (mg/capsule) Compound of the present invention 150.00 Lactose 146.15 Cornstarch 50.00 Colloidal Silicon dioxide 0.35 9:1 mixture of magnesium stearate/sodium lauryl sulfate 3.50 Total 350 Formulation example 2 Capsule Ingredients Amount (mg/capsule) Compound of the present invention 50.00 Lactose 48.96 Corn starch 16.75 Colloidal silicon dioxide 0.117 Magnesium stearate/lauryl 9:1 mixture of sodium sulfate 1.173 Total 117.00 Formulation example 3 Oral syrup Ingredients Amount (mg/ml) Inventive compound 5.00 Propylene glycol 103.80 Ethanol 39.68 Glycerol 378.00 Sorbitol 70% (w/w) aqueous solution 387.00 Cherry flavor 10.35 Hydrochloric acid Pure water adjusted to PH3.5 1000ml Formulation example 4 Tablet Ingredients Amount (mg/tablet) Compound of the present invention 150.0 Microcrystalline cellulose 132.0 Dicalcium phosphate (anhydrous) 132.0 Sodium croscarmellose
Croscarmellose ^* 22.5 Povidone K90 ^** 9.0 Magnesium Stearate 4.5 Total 450.0 * Cross-linked polymer of sodium carboxymethyl cellulose (commercially available) ** Consists essentially of linear 1-vinyl-2-pyrrolidone Synthetic polymer with a viscosity of 90 in aqueous solution expressed as K value (commercially available) Production Example 5 Tablet Ingredients Amount (mg/tablet) Compound of the invention 50.0 Microcrystalline cellulose 44.0 Dibasic calcium phosphate (anhydrous) 44.0 Sodium croscarmellose 7.5 Povidone K90 3.0 Magnesium stearate 1.5 Total 150.0 Formulation example 6 Intravenous injection Ingredients Amount (mg/ml) Compound of the present invention 50.0 Absolute ethanol 594.0 Distilled water for injection Adjusted to 1000.0 ml

Claims (1)

[Claims] 1 2-(2,4-difluorophenyl)-1,
3-bis(1H-1,2,4-triazole-1
-yl)propan-2-ol and/or a pharmaceutically acceptable salt thereof.