`Naito et al.
`
`IIIII IIIIIIII III IIIII IIIII IIIII IIIII IIIII M IIIII IIIII IIIIII II IIIII IIII
`5,620,994
`Apr. 15, 1997
`
`[lll Patent Number:
`[45] Date of Patent:
`
`US005620994A
`
`[54] AZOLYLAMINE DERIVATIVE
`
`FOREIGN PATENT DOCUMENTS
`
`[75] Inventors:
`
`Takanobu Naito; Haruhito
`Kobayashi; Hironobu Ogura; Kiyoshi
`Nagai; Tokiko Nishida; Tadashi
`Arika; Mamoru Yokoo; Satoko
`Shusse, all of Kyoto, Japan
`
`[73] Assignee:
`
`Kaken Pharmaceutical Co., Ltd.,
`Tokyo, Japan
`
`[21] Appl. No.: 532,800
`
`[22] Filed:
`
`Nov. 7, 1995
`
`[30]
`
`Foreign Application Priority Data
`
`May 10, 1993 [JP] Japan .................................... 5-132931
`
`[51]
`
`Int. CI.6 ...................... A61K 31/445; C07D 401/06;
`C07D 403/06
`[52] U.S. Cl ........................... 514/326; 514/212; 514/383;
`540/603; 546/210; 548/267.2; 548/267.8
`[58] Field of Search ............................ 540/603; 546/210;
`548/267.2, 267.8; 514/212, 326, 383
`
`[56]
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`4,507,484
`
`3/1985 Gymer et al..
`
`0054974A1 6/1982 European Pat. Off..
`2159148 11/1985 United Kingdom.
`
`Primary Examiner--Patricia L. Morris
`Attorney, Agent, or Firm--Armstrong, Westerman, Hattori,
`McLeland & Naughton
`
`[57].
`
`ABSTRACT
`
`There is disclosed a fungicide containing, effective ingredi-
`ent, a compound having the general formula (I):
`
`[I N (I)
`
`~" N OH CH3 (CH2)m R1
`
`x )
`,,1,, /
`
`)=<
`
`CH2--C--CH--N
`I *2
`\
`At"
`(CH2)n
`or an acid addition salt thereof, particularly the compound
`wherein an absolute configuration of the asymmetric carbon
`atoms is R,R-configuration or an acid addition salt thereof.
`
`R2
`
`12 Claims, No Drawings
`
`ARGENTUM EX1007
`
`Page 1
`
`
`
`5,620,994
`
`1
`AZOLYLAMINE DERIVATIVE
`
`TECHNICAL FIELD
`
`The present invention relates to an azolylamine derivative 5
`which is effective for treatment for mycosis in human and
`animals and useful as fungicides for agricultural and horti-
`cultural use or industrial use.
`
`BACKGROUND ART
`
`10
`
`Azolylamine derivatives having, in the molecule, both of
`an azolyl group such as triazolyl group or imidazolyl group
`and a cyclic amino group such as piperidino group, pyrro-
`lidino group or morpholino group are described in JP- 15
`A(Japanese Unexamined Patent Publicafion)-140768/1982
`and GB-A-2159148. However, it is hard to say in the aspect
`of an antifungal action etc. that each compound has suffi-
`cient efficacy as a medicament. Furthermore, any compound
`having methylene group or a substituted methylene group on 2o
`the cyclic amino group is not disclosed therein.
`
`The present invention provides a novel azolylamine
`derivative showing the potent antifungal activity which is
`characterized by having methylene group or a substituted
`methylene group on the cyclic amino group.
`
`25
`
`DISCLOSURE OF THE INVENTION
`
`The present invention provides a compound having the
`general formula (I):
`
`30
`
`II N (I)
`
`x )
`
`~" N OH CH3 (CH2)m Rt
`
`CH2--C--CH--N
`I *2 \
`Ar
`(CH2)n
`
`R~
`
`wherein
`Ar is non-substituted phenyl group or a phenyl group 40
`substituted with 1 to 3 subsfituents selected from a
`halogen atom and trifluoromethyl,
`Rt and R2 are the same or different and are hydrogen
`atom, a lower alkyl group, a non-substituted aryl group,
`an aryl group substituted with 1 to 3 subsfituents 45
`selected from a halogen atom and a lower alkyl group,
`an alkenyl group, an aikynyl group or an aralkyl group,
`m is 2 or 3,
`n is 1 or 2,
`X is nitrogen atom or CH, and
`*1 and *2 mean an asymmetric carbon atom,
`or an acid addition salt thereof.
`As the above-mentioned compound having the general
`formula (I), there are particularly preferable the compound 55
`wherein an absolute configuration of the asymmetric carbon
`atoms with * 1 and *2 is R,R-configurafion, and the com-
`pound being a mixture containing the compound having the
`general formula (I) wherein the absolute configuration of the
`asymmetric carbon atoms with *1 and *2 is R,R-configu- 60
`ration or an acid addition salt thereof and other optical
`isomer.
`The present invention also provides a fungicide contain-
`ing the above-mentioned compound having the general
`formula (I) or an acid addition salt thereof as an effective 65
`ingredient, and a process for treating mycosis using the
`above-mentioned compound.
`
`5o
`
`2
`BEST MODE FOR CARRYING OUT THE
`INVENTION
`
`In the above-mentioned general formula (I), the substi-
`tuted phenyl group is a phenyl group having 1 to 3 substitu-
`ents selected from a halogen atom and trifluoromethyl, and
`includes, for instance, 2,4-difluorophenyl, 2,4-dichlorophe-
`nyl, 4-fluorophenyl, 4-chlorophenyl, 2-chlorophenyl, 4-trif-
`luoromethylphenyl, 2-chloro-4-fluorophenyl, 4-bromophe-
`nyl or the like.
`The lower alkyl group includes, for instance, a straight
`chain, branched chain or cyclic alkyl group having 1 to 6
`carbon atoms such as methyl, ethyl, n-propyl, isopropyl,
`n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl,
`neopentyl and tert-pentyl.
`The non-substituted aryl group includes, for instance,
`phenyl, naphthyl, hiphenyl, or the like.
`The substituted aryl group includes, for instance, 2,4-
`difluorophenyl, 2,4-dichlorophenyl, 4-fluorophenyl, 4-chlo-
`rophenyl, 2-chlorophenyl, 4-trifluoromethylphenyl,
`2-chloro-4-fluorophenyl, 4-bromophenyl, 4-tert-butylphe-
`nyl, 4-nitrophenyl, or the like.
`The alkenyl group includes, for instance, vinyl, 1-prope-
`nyl, styryl, or the like. The alkynyl group includes, for
`instance, ethynyl, or the like.
`The aralkyl group includes, for instance, benzyl, naphth-
`ylmethyl, 4-nitrobenzyl, or the like.
`The compound of the present invention having the general
`formula (I) contains at least two asymmetric carbon atoms in
`the molecule, and there exsist an optical isomer and a
`diastereomer. With respect to the optical isomer, both enan-
`tiomers can be obtained according to the general procedure
`of optical resolution or asymmetric synthesis. A separation
`of the diastereomer can be carried out according to the usual
`separation procedure such as a fractional recrystallization or
`a chromatography to give each isomer. The compound
`having the general formula (I) includes one of these isomers
`or a mixture thereof.
`Among these, the compound wherein an absolute con-
`fguration of the asymmetric carbon atoms is R,R-configu-
`ration, has particularly potent antifungal action and therefore
`it is preferably used particularly.
`Representative examples of the compound of the present
`invention having the general formula (I) include, for
`instance,
`
`(2R,3R)-2-(2,4-difluorophenyl)-3 - (4-methylenepiperidine-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3S)-2-(2,4-difiuoropbenyl)-3-(4-methylenepiperidine-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difiuorophenyl)-3 - (4-methylenepiperi-
`dine-l-yl)-l-(1H-1,2,4-tfiazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-methylenepiperidine-
`1-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`(2S,3 S)-2- (2,4-difiuorophenyl)-3 -(4-methylenepiperidine-
`1-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-methylenepiperi-
`dine-l-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`(2R,3R)-2-(4-chlorophenyl)-3- (4-methylenepiperidine- 1-
`yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3S)-2-(4-chlorophenyl)-3-(4-methylenepiperidine- 1-
`yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(4-chlorophenyl)-3-(4-methylenepiperidine-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2- (4-ch!orophenyl)-3 -(4-methylenepiperidine- 1 -
`yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`
`Page 2
`
`
`
`5,620,994
`
`20
`
`25
`
`40
`
`45
`
`4
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-cyclohexylmethylene-
`piperdine-l-yl)-l-(1H-1,2,4?triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2- (2,4-difluorophenyl)-3-(4-cyclohexylmethyl-
`enepiperdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-benzylidenepiperdine-
`1-y!)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3S)-2- (2,4-difluorophenyl)-3 -(4-benzylidenepiperdine-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`10 (2RS,3RS)-2-(2,4-difluorophenyl)-3 -(4-benzylidenepiper-
`dine- 1-yl)- 1-(1H- 1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-isopropylidenepiper-
`dine- 1-yl)- 1-(1H- 1,2,4-triazole- 1-yl)butane-2-ol,
`
`3
`(2S,3 S )-2-(4-chlorophenyl)-3- (4-methylenepiperidine- 1-
`yl)-l-(1H-imidazole-1-yl)butane-2-01,
`(2RS,3RS)-2-(4-chlorophenyl)-3-(4-methylenepiperidine-
`1-yl)-l-(1H-imidazole-l-yl)butane-2-01,
`(2R,3R)-2-(4-trifluoromethylphenyl)-3 -(4-methylenepiperi- 5
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3 S)-2-(4-trifluoromethylphenyl)-3- (4-methylenepiperi-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2- (4-trifluoromethylphenyl)-3-(4-methylenepip-
`eridine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(4-trifluoromethylphenyl)-3- (4-methylenepiperi-
`dine-l-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`(2S,3S)-2-(4-trifluoromethylphenyl)-3-(4-methylenepiperi-
`dine-l-yl)- 1-(1H-imidazole- 1-yl)butane-2-ol,
`(2RS,3RS)-2-(4-trifluoromethylphenyl)-3-(4-methylenepip- 15 (2S,3S)-2-(2,4-difluorophenyl)-3-(4-isopropylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`eridine- 1-yl)- 1-(1H-imidazole- 1-yl)butane-2-ol,
`(2R,3R)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidine-
`(2RS,3RS)-2-(2,4-difluorophenyl)-3- (4-isopropylidenepip-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`erdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3S)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidine-
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-diphenylmethylene-
`1-yl)- 1-(1H- 1,2,4-triazole- 1-yl)butane-2-ol,
`piperdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperi-
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-diphenylmethylenepip-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`erdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidine-
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-diphenylmethyl-
`1-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`enepiperdine- 1-yl)- 1-(1 H- 1,2,4-triazole- 1-yl)butane-2-ol,
`(2S,3 S)-2-(2,4-dichlorophenyl)-3 -(4-methylenepiperidine-
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-propenylidenepiper-
`1-yl)- 1-(1H-imidazole- 1-yl)butane-2-ol,
`dine- 1-yl)- 1-(1H-1,2,4-triazole- 1-yl)butane-2-01,
`(2RS,3RS)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperi-
`dine-l-yl)-l-(1H-imidazole-l-yl)butane-2-ol,
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-propenylidenepiper-
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-ethylidenepiperdine-
`dine-l-yl)-l-(1H-1,2,4-triazole-1-yl)butane-2-01,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`30 (2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-propenylidenepip-
`(2S,3 S)-2-(2,4-difluorophenyl)-3-(4-ethylidenepiperdine- 1-
`erdine- 1-yl)- 1-(1H-1,2,4-triazole-l-yl)butane-2-01,
`yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-01,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-propynylidenepiper-
`(2RS,3RS)-2- (2,4-difluorophenyl)-3-(4-ethylidenepiper-
`dine- 1-yl)- 1-(1H- 1,2,4-triazole- 1-yl)butane-2-ol,
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-propylidenepiperdine- 35 (2S,3S)-2-(2,4-difluorophenyl)-3-(4-propynylidenepiper-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol, dine-1-yl)-l-(1H-1,2,4-triazole-1-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-propynylidenepip-
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-propylidenepiperdine-
`erdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3- (4-propylidenepiper-
`(2R,3R)-2-(2,4-difluorophenyl)-3-(3-methylenepiperidine-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-n-butylidenepiper-
`(2S,3 S)-2- (2,4-difluorophenyl)-3-(3 -methylenepiperidine-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3 S)-2-(2,4-difluorophenyl)-3-(4-n-butylidenepiperdine-
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(3-methylenepiperi-
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-n-butylidenepiper-
`(2R,3R)-2-(2,4-difluorophenyl)-3-(3-methylenepyrrolidine-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-n-pentylidenepiper-
`(2S,3 S)-2-(2,4-difluorophenyl)-3 -(3-methylenepyrrolidine-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`1-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-n-pentylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-o!,
`50 (2RS,3RS)-2-(2,4-difluorophenyl)-3-(3 -methylenepyrroli-
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-n-pentylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol, and the
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`like.
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-n-hexylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-tdazole-l-yl)butane-2-ol,
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-n-hexylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-n-hexylidenepiper-
`dine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-cyclopropylmethyl-
`ene-piperdine- 1-yl)- 1-( 1 H- 1,2,4-triazole- 1-yl)butane-2- 60
`ol,
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-cyclopropylmethylene-
`piperdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-cyclopropylmethyl-
`enepiperdine- 1-yl)- 1-(1H- 1,2,4-triazole- 1-yl)butane-2-ol, 65
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-cyclohexylmethylene-
`piperdine-l-yl)- 1-(1H-1,2,4-triazole-l-yl)butane-2-ol,
`
`The compound of the present invention having the general
`55 formula (I) can be prepared according to the process shown
`
`as below:
`
`II
`
`x
`
`>
`
`Page 3
`
`
`
`5,620,994
`
`5
`-continued
`N
`
`)
`
`[I
`
`x
`
`"~ N OH CH3 (CH2)m RI
`
`>=(
`
`CH2--C--CH--N
`
`E *2
`
`\
`
`Ar
`
`(I)
`
`(CH2)n
`
`R~
`
`15
`
`6
`A reaction temperature is room temperature to 200° C.,
`preferably 50° to 150° C. A reaction time is 1 to 72 hours.
`After the end of the reaction, the solvent is removed and
`then purification is carried out according to a procedure such
`5 as a recrystallizafion or a chromatography. Thereby the
`compound of the present invention having the general
`formula (I) is isolated.
`The compound of the present invention having the general
`formula (I) can, if required, form a pharmaceutically accept-
`able salt thereof, for example, a salt thereof with an inor-
`ganic acid such as hydrochloric acid, sulfuric acid, nitric
`acid, phosphoric acid or hydrobromic acid, and a salt thereof
`with an organic acid such as fumaric acid, maleic acid, acetic
`acid, malic acid, tartaric acid, citric acid, methanesulfonic
`acid or toluenesulfonic acid.
`Then, the antifungal activity of the compound of the
`present invention having the above-mentioned general for-
`mula (I) is described. Test compound number used in the
`following tests was referred to the example number
`20 described below.
`1. Determination of the minimum inhibitory concentration
`(MIC)
`MIC of a test compound against Candida albicans ATCC-
`10259 was determined by the both dilution method employ-
`25 ing synthetic amino acid medium (SAAMF medium).
`Namely, to 3 lal of twofold dilution series of solution
`containing the test compound was added 300 # of SAAMF
`medium inoculated with the fungus at the final concentration
`of lxl03 a cells/ml. After thus obtained mixture was incu-
`30 bated at 35° C. for 2 days, MIC was determined by exam-
`ining a minimum concentration of the test compound in
`which concentration the test compound inhibited the growth
`of the fungus. MIC of a test compound against the fungus
`other than the Candida albicans was determined by the agar
`35 dilution method employing Sabouraud’s agar medium. That
`is to say, a test compound was dissolved in dimethyl
`sulfoxide to give a solution containing the test compound in
`the concentration of 10 mg/ml. Further, thus obtained solu-
`tion was diluted with dimethyl sulfoxide according to two-
`40 fold dilution series and 0.1 ml of the diluted solution was
`taken into a sterile shale. After 9.9 ml of Sabouraud’s agar
`medium was added thereto, the mixture was sufficiently
`mixed to give a drug-added plate. The plate was inoculated
`with 5 N of a fungus suspension at 106 cells/ml by
`45 Microplanter (Sakuma Seisakusho Co., Ltd.). As to
`Aspergillus fi~migatus NI-5561 and Cryptococcus neofor-
`mans NI-7496, a plate was incubated at 30° C. for 48 hours.
`As to Trichophyton mentagrophytes KD-01, a plate was
`incubated at 30° C. for 7 days. After incubation, MIC was
`50 determined by examining a minimum concentration of a test
`compound in which concentration the test compound inhib-
`ited the growth of the fungus. The results thereof are shown
`in Table 1. Clotrimazole and fluconazole were used as
`comparative control compounds.
`55 The abbreviated designation of names of the test fungi is
`as follows:
`
`60
`
`65
`
`Name of fungus
`
`Abbreviated designation
`
`Candida albicans ATCC 10259
`Cryptococcus neoformans NI-7496
`Aspergillus furaigatus NI-5561
`Trichophyton mentagrophytes KD-01
`
`C.a.
`Cr.n.
`A.f.
`T.m.
`
`The antifungal activity (the minimum inhibitory concen-
`tration MIC) of the compound of the present invention in the
`Examples against each fungus is shown in Table 1.
`
`(In the above-mentioned formulae, At, R1, R2, X, m and n 10
`have the same meanings as defined above.)
`Namely, the reaction of an epoxy compound having the
`general formula (II) and an amine derivative having the
`general formula (III) can lead to the compound having the
`general formula (I).
`The epoxy compound having the general formula 0I) can
`be obtained according to such process as is described in
`JP-A(Japanese Unexamined Patent Publication)-19 1262/
`1990 etc., for example, a process wherein a compound
`having the general formula (IV);
`
`[I
`x
`
`N
`
`(IV)
`
`J
`
`N
`OH CH3
`I I
`I
`CH2--C--CH--OH
`I
`Ar
`wherein Ar and X have the same meanings as defined above,
`is reacted in the presence of a base with a compound havin~
`the formula R3SOz--O~SO2R3 or R3SO2--Z, wherein R°
`is a lower alkyl group, a halogenated lower alkyl group, or
`a phenyl group which may be substituted, and Z is a leaving
`group such as a halogen atom, to give a compound (V):
`
`(v)
`
`[1 N
`
`x )
`~" N OH CH3
`I I
`CH2--C--CH--OSO2R3
`
`Ar
`
`and then the compound (V) is reacted with a base.
`The amine derivative having the general formula (11I) can
`be obtained according to the known synthetic process
`described in, for example, Chem. Pharm. Bull 41 (11)
`1971-1986 (1993) or processes described in Reference
`Examples of the present invention.
`In case that the amine derivative is in a form of a salt
`thereof with an acid such as a base, the amine derivative is
`used in a form of a free amine by being neutralized previ-
`ously or in a reaction solution with an inorganic base such
`as sodium hydroxide or an organic base such as triethy-
`lamine.
`The reaction is usually carried out using water, an organic
`solvent or a mixed solution of water and an organic solvent,
`or in the absence of any solvent. As the organic solvent, a
`solvent which does not react with a starting compound can
`be used. For example, an alcohol such as methanol, ethanol,
`n-propanol, isopropanol, n-butanol, tert-butanol, ethylene
`glycol, propylene glycol, grycerin or methyl cellosolve, an
`ether such as tetrahydrofuran, dioxane or dimethoxyethane,
`an amide such as N,N-dimethylformamide or N,N-dimethy-
`lacetamide, dimethyl sulfoxide, and the like can be used
`alone or in a mixture thereof.
`In the above-mentioned reaction system, the reaction
`advances more smoothly by adding 1 to 80 v/v % of water
`in the mixed solution to the reaction system in comparison
`with using only an organic solvent.
`With respect to an amount of each material in the reaction
`solution, from 1 to 20 mol of the compound (III) is used per
`mol of the compound (II).
`
`Page 4
`
`
`
`5,620,994
`
`7
`
`TABLE 1
`
`Test
`compound
`
`Minimum inhibitory
`concentration (MIC (lag/ml)
`Test fungus
`
`(Ex. No.)
`
`C.a.
`
`Cr.n.
`
`A.f.
`
`T.m.
`
`1
`2
`3
`4
`5
`6
`7
`8
`10
`12
`13
`14
`Clotrimazole
`Flueonazole
`
`<0.025
`<0.025
`0.39
`<0.025
`<0.025
`<0.0125
`0.025
`<0.025
`<0.025
`<0.025
`0.1
`<0.025
`0.025
`0.39
`
`0.05
`0.1
`0.78
`<0.025
`0.025
`0.2
`0.05
`0.1
`0.025
`0.1
`0.39
`0.39
`0.2
`12.5
`
`0.05
`0. I
`>100
`0.05
`0.05
`6.25
`0.39
`0.2
`0.1
`0.2
`0.78
`0.39
`0.78
`>100
`
`0.39
`0.39
`50
`<0.025
`0.1
`3.13
`0.39
`0.78
`0.39
`0.78
`1.56
`0.78
`0.39
`>100
`
`Clotrimazole
`
`Fluconazole
`
`II N
`
`= )
`x. N
`OH
`I
`I
`
`N
`
`/=
`
`8
`agar medium for 7 days. Inhibitory ratio was calculated
`according to the following formula:
`Inhibitory ratio (%)={1-(number of tissue specimens
`found fungi/total number of tissue specimens)}xl00
`The results are shown in Table 2. Clotrimazole was used
`as a control compound.
`
`TABLE 2
`
`Group
`
`10
`
`Inhibitory ratio (%)
`
`Control (non-treated)
`Control (vehicle)
`Compound of Example 1
`Clotfimazole
`
`0
`0
`98
`20
`
`15
`
`(2) Therapeutic effect on cutaneous candidiasis in guinea
`pigs.
`In the back of male Hartley guinea pig, weighing 400 to
`500 g, a portion of skin was unhalred, to which 0.1 ml of
`spore suspension of Candida albicans KC-36 (5x107 cells/
`20 ml) was dropped and the skin surface was infected by
`
`rubbing it with a glass rod. To facilitate the infection,
`prednisolone was subcutaneously administered at 30 mg/kg
`on one day before the infection, the day of infection and 4
`days after the infection. The test compound was dissolved in
`25 polyethylene glycol 400-ethanol (75:25) so as to give a 1%
`
`solution thereof and 0.2 ml of the resultant solution was
`applied for treatment once a day for 3 days from 2 days after
`the infection. The animal was killed by etherization 2 days
`after the last treatment and 10 tissue specimens of skin were
`30 cut out from the infected portion and incubated on CAN-
`DIDA GS AGAR ’EIKEN’ (EIKEN CHEMICAL CO.,
`LTD.) for 7 days. The inhibitory ratio was calculated accord-
`ing to the above-mentioned formula. The results are shown
`in Table 3. Clotrimazole was employed as a control com-
`35 pound.
`
`TABLE 3
`
`Group
`
`Inhibitory ratio (%)
`
`4O
`
`Control (non-treated)
`Control (vehicle)
`Compound of Example 1
`aotrimazole
`
`4
`8
`98
`96
`
`The above-mentioned results reveal that the compound of 45
`the present invention having the general formula (I), espe-
`cially the compound wherein the absolute configuration is
`R,R-configuration, has extremely high activity in compari-
`son with conventional fungicides.
`Furthermore, compared to Clotrirnazole and fluconazole, 50
`it is found that the compound of the present invention, i.e.
`the compound wherein a cyclic amino group having meth-
`ylene group is bonded, has surprisingly high activity.
`2. Test on treatment for infection
`(1) Effect on trichophytosis in guinea pigs.
`In the back of male Hartley guinea pig, weighing 400 to
`500 g, a portion of skin was unhaired and rubbed lightly with
`sandpaper, to which 0.1 ml of microconidium suspension of
`Trichophyton mentagrophytes KD-04 (107 cells/m1) Was
`dropped and the skin surface was infected by rubbing it with 60
`a glass rod. The test compound was dissolved in polyeth-
`ylene glycol 400-ethanol (75:25) so as to give a 1% solution
`thereof and 0.2 ml of the resultant solution was applied for
`treatment once a day for 10 days from 3 days after the
`infection. The animal was killed by etherization 2 days after
`the last treatment and 10 tissue specimens of skin were cut
`out from the infected portion and incubated on Sabouraud’s
`
`55
`
`Based on the above tests 1 and 2, it was found that the
`compound of the present invention had strong and widely
`efficacious antifungal action.
`3. Acute toxicity test for mice
`The compound of Example 1 was dissolved in polyeth-
`ylene glycol 200 and the resultant solution was applied to a
`male ICR mouse of 5 weeks old by oral or subcutaneous
`administration. The results are shown in Table 4.
`
`TABLE 4
`
`Number of died mice/
`number of tested mice
`
`Dose
`
`subcutaneous
`
`I000 mg/kg
`500 mg/kg
`250 mg/kg
`125 mg/kg
`
`0/3
`0/3
`0/3
`0/3
`
`oral
`
`0/3
`0/3
`--
`--
`
`As shown in the above Table, it is found that the com-
`65 pound of the present invention has low toxicity.
`The compounds of the present invention have strong
`antifungal activity and low toxicity. A fungicide containing
`
`Page 5
`
`
`
`5,620,994
`
`9
`the compound of the present invention having the general
`formula (I) as an effective ingredient can be employed to
`treat local and generalized mycosis in a mammal including
`human, which are caused by a fungus, especially such as
`Candida, Trichophyton, Microspornm, Epidermophyton,
`Malassezia, Cryptococcus neoformans, Aspergillus, Coccid-
`ioides, Paracoccidioides, Histoplasma or Blastomyces. The
`fungicide containing the compound of the present invention
`as an effective ingredient is useful not only for treatment for
`mycosis in human and animals but also as fungicides for
`agricultural and horticultural use, fungicides for industrial
`use and the like.
`The fungicide containing the compound of the present
`invention having the general formula (I) as an effective
`ingredient may comprise the compound alone or may be a
`mixture of the compound and liquid or solid auxiliary
`ingredients in preparing a pharmaceutical preparation such
`as an excipient, a binder and a diluent. The fungicide can be
`externally applied or orally or parenterally administered. If
`required, the fungicide may contain other medicament.
`In the case of administering the compound as an extemal
`preparation, the preparation may be in a dosage form such
`as a cream, a liquid preparation, an ointment, an oculentum,
`a suppository, a vaginal suppository, a powder or an emul-
`sion. In preparing the external preparation, there can be used
`an oily base, an emulsion base or the like. A preferable
`content of the effective ingredient is 0.1 to 10% by weight.
`The dosage may suitably vary with an area of an affected
`part and the symptom.
`In case of oral administration, the fungicide is used as a
`powder, a tablet, a granule, a capsule or a syrup, and further,
`the fungicide is also used as a injection such as a subcuta-
`neous injection, an intramuscular injection or an intravenous
`injection.
`Although the dosage is different according to the age and
`body weight of a patient and an individual condition, the
`dosage is 10 mg to 10 g, preferably about 50 mg to about 5
`g as an effective ingredient per day for an adult. With respect
`to a manner of administration, the compound is administered
`at the above-mentioned dosage per day in one to several
`times.
`The present invention is more specifically explained by
`means of the following Examples and Reference Examples.
`However, it is to be understood that the present invention is
`not limited to those Examples.
`1H-NMR spectra were determined in the solution of
`deuteriochloroform (CDC13) using tetramethylsilane as an
`internal standard by means of JNM-EX270 spectrometer
`(JEOL LTD.), and a value of chemical shift (8) was
`expressed with ppm. The determination by high perfor-
`mance liquid chromatography (hereinafter, referred to as
`HPLC) was carded out using an chiral column, CHIRAL-
`CEL OJ (4.6 minx25 cm, Daicel Chemical Industries, Ltd.)
`by means of LC-6A (HPLC apparatus, Shimadzu Corpora-
`tion).
`
`EXAMPLE 1
`
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-methylenepip-
`erdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-01
`
`10
`was removed therefrom. To the residue there were added 3
`ml of ethanol, 251 mg of (2R,3S)-2-(2,4-difluorophenyl)-3-
`methyl-2-[(1H-1,2,4-triazole-l-yl)methyl]oxirane and 3 ml
`of distilled water in order, and the mixture was refluxed with
`5 heating for 24 hours in the oil bath at 85° C. After the
`reaction, the reaction solution was cooled to room tempera-
`ture, and thereto were added 20 mi of ethyl acetate and 20
`ml of distilled water, and the organic phase was separated.
`The aqueous phase was further extracted with 10 ml of ethyl
`10 acetate, and the organic phase was combined with the
`above-separated organic phase, and the mixture was washed
`with a saturated aqueous solution of sodium chloride, and
`dried over anhydrous magnesium sulfate and then the sol-
`vent was removed. The residue was subjected to HPLC
`15 using 8 g of silica gel and was eluted with a mixed solvent
`of ethyl acetate/hexane (4:1 to 3:1) to obtain 188 mg of the
`titled compound. Yield: 54.0%. Upon recrystallization from
`a mixed solvent of ether/hexane, a pure product having a
`melting point of 860-87° C. was obtained.
`HPLC: The analysis was carried out using hexane/isopro-
`pyl alcohol of 9/1 as a mobile phase, at a flow rate of 1.0
`ml/min at room temperature under the conditions capable of
`detecting with UV (254 rim), and then a single peak
`appeared at a retention time of 6.6 minutes.
`Specific rotation: [a]o2S-93° (C=I.00, CHC13)
`Elemental analysis: For C18H22F2N40 Calculated:
`C,62.15; H, 6.36; N, 16.02 Found: C, 62.05; H, 6.37; N,
`16.08
`
`25
`
`20
`
`30 1H-NMR spectrum (CDC13) ~ ppm: 0.96 (3H,dd), 2.1-2.5
`(6H,m), 2.6-2.8 (2H,m), 2.91 (1H,q), 4.64 (2H,s), 4.80
`(1H,d), 4.89 (1H,d), 5.48 (1H,brs), 6.7-6.8 (2H,m),
`7.47-7.63 (1H,m), 7.79 (1H,s), 8.03 (1H,s)
`
`35
`
`EXAMPLE 2
`
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-methylene-
`piperdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-01
`
`The rifled compound was obtained in the same manner as
`40 in Example 1 except that instead of (2R,3S)-2-(2,4-difluo-
`rophenyl)-3-methyl-2-[(1H- 1,2,4-triazole- 1-yl) methyl]ox-
`irane, (2RS,3 SR)-2-(2,4-difluorophenyl)-3-methyl-2-[(1H-
`1,2,4-triazole- 1-yl)methyl]oxirane being a racemic
`modification thereof was used.
`HPLC: The analysis was carried out using hexane/isopro-
`pyl alcohol of 911 as a mobile phase, at a flow rate of 1.0
`ml/min at room temperature under the conditions capable of
`detecting with UV (254 nm), and then two peaks having an
`area ratio thereof of 1:1 appeared at retention times of 6.6
`50 minute and 5.8 minute, respectively.
`
`45
`
`XH-NMR spectrum (CDC13) 8 ppm: 0.96 (3H,dd, J-3
`Hz,7 Hz), 2.1-2.5 (6H, m), 2.6-2.8 (2H,m), 2.91 (1H,q,J=7
`Hz), 4.64 (2H,s), 4.80 (1H,d,J=15 Hz), 4.89 (1H,d,l=15 Hz),
`5.47 (1H,brs), 6.%6.8 (2H,m), 7.5-7.6 (1H,m), 7.79 (1H,s),
`8.02 (1H,s)
`
`EXAMPLE 3
`
`(2S, 3 S ) -2- (2,4- difluorophenyl)- 3 - (4-methylene-pip-
`erdine-l-yl)-l-(1H-1,2,4-triazole-l-yl)butane-2-ol
`
`55
`
`60
`
`There was added 11.2 ml of 50% aqueous solution of
`potassium hydroxide to 1.336 g of 4-methylenepiperidine
`hydrochloride and, after dissolved under stirring, the result-
`ing solution was extracted with 20 rnl of ethyl ether. Then 65
`the aqueous phase was further extracted with 10 ml of ethyl
`ether, and the organic phases were combined and ethyl ether
`
`The rifled compound was obtained in the same manner as
`in Example 1 except that instead of (2R,3S)-2-( 2,4-difluo-
`rophenyl)-3-methyl-2-[(1H-1,2,4-triazole-l-yl)methyl]ox-
`irane, (2S,3R)-2-(2,4-difluorophenyl)-3-methyl-2-[(1H-1,2,
`4-triazole-l-yl)methyl]oxirane being an enantiomer thereof
`was used.
`
`Page 6
`
`
`
`5,620,994
`
`11
`HPLC: The analysis was carried out using hexane/isopro-
`pyl alcohol of 9/1 as a mobile phase, at a flow rate of 1.0
`ml/min at room temperature under the conditions capable of
`
`detecting with UV (254 rim), and then a single peak 5
`appeared at a retention time of 5.8 minutes.
`
`1H-NMR spectrum (CDC13) 6 ppm: 0.96 (3H,dd,J-3 Hz,7
`Hz), 2.1-2.5 (6H,m), 2.6-2.8 (2H,m), 2.91 (1H,q,J=7 Hz),
`
`4.64 (2H,s), 4.80 (1H,d,J=15 Hz), 4.89 (1H,d,J=15 Hz), 5.48 10
`(1H,brs), 6.7-6.8 (2H,m), 7.5-7.6 (1H,m), 7.78 (1H,s), 8.03
`
`0H,s)
`
`15
`
`EXAMPLES 4 TO 14
`
`The compounds of Examples 4 to 14 shown in Table 6
`were synthesized using starting materials shown in Table 5
`in the same manner as in Example 1.
`
`20
`
`12
`
`TABLE 5
`
`Epoxy compound (II)
`
`Amine derivative (IID
`
`[I
`X
`
`N
`
`"-N "3
`
`o
`
`I "1 / \
`CH2--C
`CH--CH3
`1
`*2
`Ar
`
`/ (CH2)m
`
`\
`
`(CH2).
`
`R1
`
`R2
`
`Ex.
`No.
`
`4
`5
`6
`7
`8
`9
`10
`11
`12
`13
`14.
`
`X
`
`*1 *2
`
`Ar
`
`R1
`
`R2
`
`m n
`
`CH R
`S 2,4-difluorophenyl H
`CH RS SR 2,4-difluorophenyl H
`N
`R
`S 4-chlorophenyl
`H
`N
`R
`S 2,4-dichlorophenyl H
`N
`RS SR 2,4-difluorophenyt H
`N
`R
`S 2,4-difluorophenyl H
`N
`RS SR 2,4-difluorophenyl H
`N
`R
`S 2,4-difluorophenyl Ph*
`R
`S 2,4-