illlllllllfllllgilllllllllllllllillllllUlllllllllllllllllllll
`
`US005716969A
`
`United States Patent
`Naito et al.
`
`[19]
`
`[11] Patent Number:
`
`[45] Date of Patent:
`
`5,716,969
`Feb. 10, 1998
`
`[54]
`
`[75]
`
`AZOLYLAMINE DERIVATIVE
`
`[56]
`
`References CRed
`
`Inventors: Takanobu Naito; Haruldto
`Kobayashi; Hironobu Ogura; Kiyoshi
`Nagai; Tokiko N’mhida; Tadashi
`Arika; Mamoru Yokoo; Satoko
`Shusse, all of Kyoto, Japan
`
`U.S. PATENT DOCUMENTS
`
`4,507,484 3/1985 Gymex et al.
`
`FOREIGN PATENT DOCUMENTS
`
`0 054 974 AI 6/1982 European Pat. Off..
`2 159 148 11/1985 United Kingdom
`
`[73] Assignee: Kaken Pharmaceutical Co., Ltd.,
`Tokyo, Japan
`
`[21] AppL No.: 781,204
`
`[22] Filed:
`
`Jan. 9, 1997
`
`Related U.S. Application Data
`
`[62] Division of Set. No. 532,800, filed as PCT/JP94/00737, May
`2, 1994, Pat. No. 5,620,994.
`
`[30] Foreign Application Priority Data
`
`May 10, 1993 [JP] Japan ....................................... 132931
`
`[51]
`
`Int. 0.6 ...................... A61K 31/445; C07D 401/06;
`C07D 403/06
`[52] U.S. CI ........................... 514/326; 540/603; 546/210;
`548/314.7; 514/212; 514/397
`[58] Field of Search ......................... 546/210; 548/314.7;
`514/326, 397, 212; 540/603
`
`Primary Examiner--Patricia L. Morris
`Attorney, Agent, or Firm---Armstrong, Westerman, Hattori,
`McLeland & Naughton
`
`[57]
`
`ABSTRACT
`
`There is disclosed a fungicide containing, as an effective
`ingredient, a compound having the general formula (I):
`
`[I
`
`X
`
`CH3
`N ~ OH
`/(CH2).
`I
`I
`I
`CH=--’IC--CII--N
`>===~
`
`R1
`
`Ar
`
`(OH2).
`
`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.
`
`9 Claims, No Drawings
`
`ARGENTUM EX1008
`
`Page 1
`
`

`

`1
`AZOLYLAMINE DERIVATIVE
`
`5,716,969
`
`This is a division of application Ser. No. 08/532,800 filed
`Nov. 7, 1995, which is a U.S, national stage under §371 of
`application No. PCT/JtXM/00737 filed May 2. 1994, claim- 5
`ing priority from Japanese patent application No. 132931
`filed May 10, 1993.
`
`2
`ingredient, and a process for treating mycosis using the
`above-mentioned compound.
`
`BEST MODE FOR CARRYING OUT THE
`INVENTION
`
`In the above-mentioned general formula (I), the substi-
`tuted phenyl group is a phenyl group having I to 3 substitu-
`ents selected from a halogen atom and trifluoromethyl, and
`includes, for instance, 2,4-difluorophenyl, 2,4-
`10 dichlorophenyl, 4-fluorophenyl, 4-chlorophenyl,
`
`3O
`
`2-chlorophenyl, 4-trifluoromethylphenyl, 2-chloro-4-
`fluorophenyl, 4-bromophenyl or the like.
`The lower alkyl group includes, for instance, a straight
`15 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,
`20 phenyl, naphthyl, biphenyl, or the like.
`The substituted aryl group includes, for instance, 2,4-
`difluorophenyl, 2,4-dichlorophenyl, 4-fluorophenyl,
`4-chiorophenyl, 2-chlorophenyl, 4-trifluoromethylphenyl,
`2-chloro-4-fluorophenyl, 4-bromophenyl, 4-tert-
`25 butylphenyl, 4-nitrophenyl, or the like.
`The alkenyl group includes, for instance, vinyl,
`1-propenyl, styryl, or the like.
`The alkynyl group includes, for instance, ethynyl, or the
`llke.
`The aralkyl group includes, for instance, benzyl,
`naphthylmethyl, 4-nitrobenzyl, or the like.
`The compound of the present invention having the general
`formula (1) contains at least two asymmetric carbon atoms in
`35 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
`4o separation procedure such as a fractional recrystalllzation 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-
`45 figuration of the asymmetric carbon atoms is R,R-
`configuration, has particularly potent antifungai action and
`therefore it is preferably used particularly.
`Representative examples of the compound of the present
`invention having the genera formula (I) include, for
`5o instance,
`(2R.3R)-2-(2,4-difluorophenyl)-3-(4-methylenepiperidino-
`1-(1H- 1,2.4-trlazol- 1 -yl)butane-2-ol,
`(2S.3 S)-2-(2.4-dittuorophenyl)-3-(4-methylenepiperidino)-
`1-(1H- 1,2.4-triazol-l-yl)butane-2-ol,
`55 (2RS,3RS)-2-(2,4-difl uorophenyl)-3-(4-
`methylen epiperidino)- 1-( 1 H- 1.2,4-triazol- 1-yl)butan-2-
`ol,
`(2R.3R)-2- (2.4-diflu orophenyl)-3 -(4-methylenepiperidino)
`-1-(1H-imidazol- 1-yl)butan-2-ol.
`6o (2S,3S)-2-(2A-difluorophenyl)-3-(4-methylenepiperidino)-
`1-yl)-l-(1H-imidazol- 1-yl)butan-2ool,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-
`methylenepiperidino)- 1 -( 1H-imidazol- 1-yl)butan-2-ol,
`(2R,3R)-2-(4-chlorophenyl)-3-(4-methylenepiperidino)- 1-
`65 ( 1 H- 1,2,4-triazol- 1 -yl)butan-2-ol.
`(2S,3 S)-2-(4-chlorophenyl)-3-(4-methylenepiperidino)- 1-
`( 1 H- 1,2,4-triazol- 1 -yl)butan-2-ol.
`
`TECHNICAL FIEI D
`
`The present invention relates to an azolylamine which is
`effective for treatment for mycosis in human and animals
`and useful as fungicides for agricultural and horticultural use
`or industrial use.
`
`BACKGROUND ART
`
`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-A
`(Japanese Unexamined Patent Publication)-140788t1982
`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
`the cyclic amino group is not disclosed therein.
`The present invention provides a novel azolylarnine
`derivative showing the potent anfifungal activity which is
`characterized by having methylene group or a substituted
`methylene group on the cyclic amino group.
`
`DISCLOSURE OF THE INVENTION
`
`The present invention provides a compound having the
`general formula (1):
`
`II
`
`N
`
`(1)
`
`, ,, / >==<
`
`N OH CH3 (CH2)m
`
`CH2--*IC
`I
`
`Ar
`
`CH--N
`"2
`\
`
`(CH2).
`
`Rt
`
`R2
`
`wherein Ar is non-substituted phenyl group or a phenyl
`group substituted with 1 to 3 substituents selected from a
`halogen atom and trifluoromethyl,
`Ra and R2 are the same or different and are hydrogen atom,
`a lower al.kyl group, a non-substituted aryl group, an aryl
`group substituted with 1 to 3 substituents selected from a
`halogen atom and a lower aikyl group, an alkenyl group,
`an alkynyl group or an aralkyl group.
`mis 2 or 3,
`his 1 or2,
`X is nitrogen atom or CH, and
`*1 and *2 mean an asymmetric carbon atom,
`or an acid addition sat thereof.
`As the above-mentioned compound having the genera
`formula (1), there are particularly preferable the compound
`wherein absolute configuration of the asymmetric carbon
`atoms with *1 and *2 is R,R-configuration, and the com-
`pound being a mixture containing the compound having the
`genera formula (1) wherein the absolute configuration of the
`asymmetric carbon atoms with *1 and *2 is R.R-
`configuration 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
`
`Page 2
`
`

`

`5,716,969
`
`3
`(2RS.3RS)-2-(4-chlorophenyl)-3-(4-methylenepiperidino)-
`1-(1H- 1 2.4-triazol- 1 -yl)butan-2-ol,
`(2R.3R)-2-(4-chlorophenyl) -3 -(4-methylenepiperidino)- 1 -
`(1H-imidazol- 1-yl)butan-2-ol,
`(2S.3 S)-2-(4-chlorophenyl)-3-(4-methylenepiperidino) - 1-
`(iH-imidazol- 1-yl)butan-2-ol.
`(2RS.3RS)-2-(4-chlorophenyl)-3-(4-methylenepiperidino)-
`1-(1H-imidazol-l-yl)butan-2-ol,
`(2R,3R)-2-(4-trifluoromethylphenyl)-3-(4-
`methylenepiperidino)- 1-(1H- 1.2,4-triazol- 1-yl)butan-2-
`ol,
`(2S.3S)-2-(4-trifl uoromethylphenyl)-3-(4-
`methylenepiperidino)- 1-(1H- 1.2.4-triazol- 1-yl)butan-2-
`ol.
`(2RS,3RS)-2- (4-trifluoromethylphenyl)-3-(4-
`methylenepiperidino)-l-(1H-1.2,4-triazol-l-yl)butan-2-
`ol,
`(2R.3R)-2-(4-trifluoromethylphenyl)-3-(4-
`methylenepiperidino)- 1-(1H-imidazol- 1-yl)butan-2-ol,
`(2 S,3S)-2- (4-trifluoromethylphenyl)-3-(4-
`methylenepiperidino)- 1-(1H-imidazol- 1 -yl)butan-2-ol.
`(2RS.3RS)-2-(4-trifluoromethylphenyl)-3- (4-
`methylenepiperidin o)- 1-( 1H-imidazol- 1 -yl)butan-2-ol,
`(2R.3R)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidino)-
`1-(1H- 12.4-triazol- 1-yl)butan-2-ol.
`(2S.3S)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidino)-
`1-(1H-1.2.4-txiazol-l-yl)butan-2-ol.
`(2RS.3 RS)-2- (2.4- dichtorophenyl)-3-(4-
`methyle n epiperidino )- 1-( 1H- 1,2.4-triazol- 1-yl)butan-2-
`ol.
`(2R3R)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidino)-
`1-(1H-imidazol- l-yl)butan-2-ol.
`(2S.3S)-2-(2,4-dichlorophenyl)-3-(4-methylenepiperidino)-
`1-( 1H-imidazol- 1-yl)butan-2-ol.
`(2RS.3RS)-2-(2,4-dichlorophenyl)-3-(4-
`methylenepiperidino)- 1-(1H-imidazol- 1-yl) butan-2-ol,
`(2R,3R)-2-(2.4-difluorophenyl)-3-(4-ethylidenepiperidino)-
`1-(1H- 1.2,4-triazol-l-yl)butan-2-ol,
`(2S.3S)-2-(2,4-difluorophenyl)-3-(4-ethylidenepiperidino)-
`1-(1H- 1.2,4-triazol-l-yl)butan-2-ol.
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-
`ethylidenepiperidino)- 1-( 1H- 1.2.4-triazol- 1-yl)butan-2-
`ol.
`(2R,3 R)- 2-(2.4- difluorophenyl)-3- (4-
`propyHdenepiperidino)- I-(1H- 1.2.4-triazol- 1-yl)butan-2-
`ol.
`(2S.3S)-2-(2.4Mifluorophenyl)-3-(4-propylidenepiperidino)
`- 1-(1H-1.2.4-triazol- 1-yl)butan-2-ol,
`(2RS.3RS)-2-(2,4-difluorophenyl)-3-(4-
`propylidenepiperidino)- 1 -( 1 H- 1.2.4-triazol- 1 -yl)butan-2 -
`ol,
`(2R.3 R)- 2- (2,4-difluorophenyl)-3-(4- n-
`butylidenepiperidino)- 1-(1H- 1,2.4-triazol- 1-yl)butan-2-
`ol,
`(2S,3 S)-2-(2,4- cliff uorophenyl)-3-(4-n-
`butylidenepiperidino)- 1-(1H- 1,2.4-triazol- 1-yl)butan-2-
`ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-n-
`butylidenepiperidino)- 1-(1H- 1.2.4-triazol- 1-yl)butan-2-
`ol.
`(2R,3R)-2-(2,4-difluorophenyl)-3-(4-n-
`pe ntylidenepiperidino)- 1-( 1H- 1.2.4-triazol- 1-yl)butan-2-
`ol,
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-n-
`pentylidenepiperidino)- 1-(1H- 1,2,4-triazol- 1-yl)butan-2-
`ol,
`(2RS.3RS)-2-(2,4-difluorophenyl)-3-(4-n-
`pentylidenepiperidino)- 1-(1H- 1,2,4-triazol- 1-yl)butan-2-
`ol,
`
`5
`
`20
`
`4
`(2R.3 R)-2- (2.4- diflu orophenyl)-3- (4- n-
`hexylidenepiperidin o)- 1 -( 1H- 1.2,4-triazol- 1 -yl)butan-2-
`ol.
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-n-
`h exylidenepiperidino)- 1 -( 1H- 1.2,4-triazol- 1 -yl)butan-2-
`ol,
`(2RS,3 RS)-2-(2,4-difl uorophenyl)-3- (4-n-
`hexylidenepiperidino)- 1-(1H- 1,2,4-triazol- 1-yl)butan-2-
`ol,
`10 (2R.3R)- 2-(2.4-difl uor ophenyl)-3-(4-
`cyclopropylmethylenepiperidino)- 1-(1H-1.2,4-triazol- 1-
`yl)butan-2-ol,
`(2 S,3 S)-2- (2,4-difluorophenyl)-3-(4-
`cyclopropylmethylenepiperidino)- 1-(1H-1.2,4-triazol- 1-
`is yl)butan-2-ol,
`(2RS,3 RS)-2-(2.4-difl u orophe nyl)-3-(4-
`cy el opropylmethylenepiperidino ) - 1-( 1H-1.2,4-triazol- 1-
`yl)butan-2-ol,
`(2R.3R)-2- (2.4-difluorophenyl)-3-(4-
`cyclohexylmethylenepiperidino)- 1-(1H- 1,2,4-triazol- 1-
`yl)butan-2-ol,
`(2 S.3S)-2-(2,4-diflu orophenyl)-3-(4-
`cyclohexylmethylenepiperidino)-l-(1H-1.2,4-triazol-1-
`yl)butan-2-ol.
`2s (2RS.3R S)-2-(2,4-difluorophenyl)-3- (4-
`cyclohexylmethylenepiperidino)-l-(1H-1,2.4-triazol-1-
`yl)butan-2-ol,
`(2R.3 R)- 2- (2,4- dill uor oph e nyl)-3- (4-
`benzylidenepiperictino)- 1-(1H- 1.2.4-triazol-l-yl)butan-2-
`30 oL
`(2S 3S)-2-(2,4-difluorophenyl)-3-(4-benzylidenepiperidino)
`-1-(1H-1.2.4-triazol-l-yl)butan-2-oL
`(2RS,3RS),2-(2,4-difl u orophenyl)-3-(4-
`benzylidenepiperidino)- 1-(1H- 1,2,4-triazol-l-yl)butan-2-
`35 ol,
`(2R,3R)-2,-(2,4-difl uorophenyl)-3-(4-
`isopropylidenepiperidino)- 1-( 1 H- 1.2,4-triazol- l-yl)
`butan-2-ol,
`(2S,3 S)-2- (2.4-difluorophenyl)-3-(4-
`isopropylidenepiperidino)- 1-(1H-1.2,4-triazol- 1-yl)
`butan-2-ol,
`(2RS,3RS)-2-(2,4-diflu orophenyl)-3-(4-
`isopropylidenepiperidino)- 1-( 1H-1.2,4-triazol- 1-yl)
`butan-2-ol,
`45 (2R.3R)-2- (2,4-difluorophenyl)-3-(4-
`diphenylmethylen epiperidino)- 1-( 1H-1,2,4-triazol- 1-yl)
`butan-2-ol,
`(2S,3S)-2- (2.4-difluorophenyl)-3-(4-
`diphenylmethylenepiperidino)- 1-(1H- 1,2,4-triazol- 1-yl)
`5o butan-2-ol,
`(2RS,3RS)- 2-(2,4-diflu oroph enyl)- 3-(4-
`diphenylmethylenepiperidino)- 1-(1H- 1,2,4-triazol- 1 -yl)
`butan-2-ol,
`(2R,3R)-2- (2,4-difluorophenyl)-3-(4-
`propenylidenepiperidino )- 1-(1H- 1,2,4-triazol- l-yl)butan-
`2 -oL
`(2S.3S)-2-(2,4-difluorophenyl)-3-(4-
`propen ylidenepiperidino)- 1-( 1 H- 1.2,4-triazol- 1 -yl)butan-
`2 -ol.
`60 (2RS,3RS),2-(2,4-difl uorophenyl)-3- (4-
`propenylidenepiperidino)- 1-(1H- 1,2,4-triazol-l-yl)butan-
`2-ol.
`(2R,3R)-2- (2,4-difluorophenyl)-3-(4-
`propenylidenepiperidino)- IH- 1,2.4-triazol-1 -yl)butan-2-
`65 ol,
`(2S .3 S)-2-(2,4-diltuorophenyl) -3 - (4-propylidenepiperidin o)
`- 1 -(1H- 1,2.4-triazol-1 -yl)butan-2-ol.
`
`4o
`
`Page 3
`
`

`

`6
`
`II
`
`N
`
`N OH CH3
`I I I
`CHz--C--CH-- OSO2R3
`I
`Ar
`
`(v)
`
`5,716,969
`
`5
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(4-
`propynylidenepiperidinol) - 1-( 1 H- 1.2.4-triazol- 1-yl)
`butan-2-ol,
`(2R.3R)-2-(2,4-difluorophenyl)-3-(3-methylenepiperidino)-
`1-(1H- 1,2,4-triazol- 1-yl)butan-2-ol, 5
`(2S,3S)-2-(2,4-difluorophenyl)-3-(3 -methylenepiperidino)-
`1-(1H- 1,2,4-triazol- 1-yl)butan-2-ol,
`(2RS,3 R S)-2-(2,4-difl uorophenyl)-3- (3-
`methylenepiperidino)- 1-(1H- 1,2,4-triazol- 1-yl)butan-2- 10
`
`oL
`(2R3R)-2-(2A-difluorophenyl)-3-(3-methylenepyrrolidino)
`-1-(1H-2.4-triazol-l-yl)butan-2-oL
`(2S.3S)-2-(2.4-difluorophenyl)-3-(3-methylenepyrrolidino) t5
`- 1-(1 H- 1.2.4-triazol- 1-yl)butan-2-ol,
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-(3-
`methylenepyrr olidino)- 1-( 1H- 1,2,4-triazol- 1-yl)butan-2-
`ol, and the like.
`The compound of the present invention having the general 20
`formula (I) can be prepared according to the process shown
`as below:
`
`(c~2),,
`X~N "~ 0 HN
`I *’ / \ rm~
`CH2--C I CJ’I ICH3
`[ ,z
`Ar
`(m
`
`II
`
`N
`
`)=<
`
`Rt 35
`
`R2
`
`and then the compound (V) is reacted with a base.
`The amine derivative having the general formula (B!) 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 fore 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,
`25 n-propanol, isopropanol, n-butanol, tert-butanol, ethylene
`glycol, propylene glycol, glycerin or methyl cellosolve, an
`ether such as tetrahydrofuran, dioxane or dimethoxyethane,
`an amide such as N,N-dimethylformamide or N.N-
`dimethylacetamide, dimethyl sulfoxide, and the like can be
`30 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 (HI) is used per
`tool of the compound (ID.
`A reaction temperature is room temperature to 200° C.,
`preferably 50° to 150° C. A reaction time is 1 to 72 hours.
`40 After the end of the reaction, the solvent is removed and
`then purification is carried out according to a procedure such
`as a recrystallization 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 sait thereof
`50 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 antifungai activity of the compound of the
`present invention having the above-mentioned general for-
`55 mula (I) is described. Test compound number used in the
`following tests was referred to the example number
`described below.
`1. Determination of the minimum inhibitory concentration
`(MIC)
`60 MIC of a test compound against Candida albicans ATCC-
`10259 was determined by the both dilution method employ-
`wherein Ar and X have the same meanings as defined above,
`ing synthetic amino acid medium (SAAMF medium).
`is reacted in the presence of a base with a compound having
`Namely, to 3 I11 of twofold dilution series of solution
`the formula R3SO2----O------SO2R3 or R3SO2--Z, wherein R3
`containing the test compound was added 300 pl of SAAMF
`is a lower aikyl group, a halogenated lower alkyl group, or 65 medium inoculated with the fungus at the final concentration
`3
`of 1 xl0 cells/ml. After thus obtained mixture was incubated
`a phenyl group which may be substituted, and Z is a leaving
`group such as a halogen atom, to give a compound (V):
`at 35° C. for 2 days, MIC was determined by examining a
`
`N
`
`OH CH3 i(CH2)m
`
`Crh--c--CH--N
`[ *z
`\
`Ar
`(CHA,
`(I)
`
`(In the above-mentioned formulae, At, R1, R2, X, m and n
`have the same meanings as defined above.)
`
`Namely. the reaction of an epoxy compound having the
`general formula (11) and an amine derivative having the
`general formula (HI) can lead to the compound having the
`general formula (I).
`
`45
`
`The epoxy compound having the general formula (11) can
`be obtained according to such process as is described in
`JP-A(Japanese Unexamined Patent Publication)-191262I
`1990 etc., for example, a process wherein a compound
`having the general formula (IV):
`
`(IV)
`
`N
`
`IJ
`x~ ~1
`N OH CH3
`I I I
`CH2 --C --CH-- OH
`I
`Ar
`
`Page 4
`
`

`

`5,716,969
`
`7
`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
`dilution method employing Sabourand’s agar medium. That 5
`is to say, a test compound was dissolved in dimethyl
`sulfoxide to give a solution containing the test compound in
`the concentration of 10 rng/rni. Further, thus obtained solu-
`tion was diluted with dimethyl suifoxide according to two- 10
`fold dilution series and 0.1 rnl of the diluted solution was
`taken into a sterile shale. After 9.9 rnl of Sabouraud’s agar
`medium was added thereto, the mixture was sut~ciently
`mixed to give a drug-added plate. The plate was inoculated
`with 5 lal of a fungus suspension at 10%elis/ml by 15
`Microplanter (Sakuma Seisakusho Co., Ltd.). As to
`Aspergillus fumigams NI-5561 and Cryptococcus neofor-
`marts NI-7496, a plate was incubated at 30° C. for 48 hours.
`As to Trichophyton mentagrophytes KD-01, a plate was 20
`incubated at 30° C. for 7 days. After incubation, MIC was
`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 25
`comparative control compounds.
`The abbreviated designation of names of the test fungi is
`as follows:
`
`8
`
`TABLE 1-continued
`
`Test
`compound
`
`Minlmttm inhibitory concentration (MIC (og/ml)
`Test fungus
`
`(Ex. No.)
`
`Clotrimazok~
`F1uconazole
`
`C .a.
`
`0.025
`0.39
`
`Cr, n,
`
`0.2
`12,5
`
`A.f.
`
`T.m.
`
`0.78
`>100
`
`0.39
`>100
`
`C lotrimazole
`
`Fluconazol¢
`N
`
`II Nx ,,21
`N OH
`
`CH2--C--CH2--N
`
`Name of fimgus
`
`Abbreviated designation
`
`3O
`
`F
`
`Candida a/b/cans ATCC 10259
`
`Cryplococcus neofommns NI-7496
`
`Aspergillusfumigatus NI-5561
`
`Trichop~ ratmagrophy~s KI)-O1
`
`C.a.
`
`Cr.n.
`
`A.f.
`
`Tan.
`
`The above-mentioned results reveal that the compound of
`35 the present invention having the general f~nnula (I), espe-
`
`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.
`
`TABLE 1
`
`40
`
`45
`
`Test
`
`compotmd
`
`Minlmmn inhibitory concentration (MIC (IagJml)
`
`Test fimgus
`
`(Ex, No.)
`
`C.a.
`
`Cry.
`
`A.f.
`
`1
`
`2
`
`<0.025
`
`<0.025
`
`0.05
`
`0.1
`
`0.05
`
`0.1
`
`Tan,
`
`0.39
`
`0.39
`
`cially the compound wherein the absolute configuration is
`R,R--configuration, has extremely high activity in compari-
`son with conventional fungicides.
`Furthermore, compared to Clotrimazole and fluconazole,
`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 triehophytosis in guinea pigs.
`In the back of male Hattley guinea pig, weigifing 400 to
`500 g, a portion of skin was unhaired and rubbed lightly with
`sandpaper, to which 0.1 ml of microconidium suspension of
`50 Trichophyton mentagrophytes KD-04 (107 cells/ml) was
`dropped and the skin surface was infected by rubbing it with
`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 rnl 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
`6o agar medium for 7 days. Inhibitory ratio was calculated
`according to the following formula:
`
`55
`
`Inhibitory ratio (%)={ 1-(number of tissue specimens found
`fungi/total number of tissue specimens)}xl00
`
`65
`
`The results are shown in Table 2. Clotrimazole was used
`as a control compound.
`
`3
`
`4
`
`5
`
`6
`
`7
`
`8
`
`10
`
`12
`
`13
`
`14
`
`0.39
`
`<0.025
`
`<0.025
`
`<0.0125
`
`0.025
`
`<0.025
`
`<0.025
`
`<0.025
`
`0.1
`
`<0.025
`
`0.78
`
`>100
`
`<0.025
`
`0.025
`
`0.2
`
`0.05
`
`0.1
`
`0.025
`
`0.1
`
`0.39
`
`0.39
`
`0.05
`
`0.05
`
`6.25
`
`0.39
`
`0,2
`
`0.1
`
`0.2
`
`0.78
`
`0.39
`
`50
`
`<0.025
`
`O. 1
`
`3.13
`
`0.39
`
`0.78
`
`0.39
`
`0.78
`
`1.56
`
`0.78
`
`Page 5
`
`

`

`5,716,969
`
`9
`
`TABLE 2
`
`Group
`
`Inhibitory ratio (%)
`
`Control (non-treated)
`Control (vehicle)
`Compound of Example I
`CloWimazol¢
`
`0
`0
`98
`20
`
`(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 unhaired, to which 0.1 ml of
`spore suspension of Candida albicans KC-36 (5x107 cells/
`nil) 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
`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
`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-
`pound.
`
`TABLE 3
`
`Group
`
`Inhibitory ratio (%)
`
`Control (non-treated)
`Control (vehic kQ
`Compotmd of Example I
`Clotrimazole
`
`4
`8
`98
`96
`
`Based on the above tests 1 and 2, it was found that the
`compound of the present invention had strong and widely 40
`etficacious anfifungal 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 45
`administration. The results are shown in Table 4.
`
`10
`Coccidioides, Paracocddioides, Histoplasma or Blastomy-
`ces. 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
`5 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
`10 mixture of the compound and Hquid 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.
`15 In the case of administering the compound as an external
`preparation, the preparation may be in a dosage form such
`as a cream, a liquid preparation, an ointment, an oculenturm
`a suppository, a vaginal suppository, a powder or an emul-
`sion. In preparing the external preparation, there can be used
`20 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
`25 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
`30 body weight of a patient and an individual condition, the
`dosage is 10 nag 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-mentioued dosage per day in one to several
`35 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.
`IH-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 (5) was
`expressed with ppm. The determination by high perfor-
`mance liquid chromatography (hereinafter. referred to as
`ItPLC) was carried out using an chiral column, CI-IIRAL-
`CEL OJ (4.6 mm~5 cm, Daicel Chemical Industries, Ltd.)
`by means of LC-6A (HPLC apparatus, Shimadzu
`Corporation).
`
`50
`
`EXAMPLE 1
`(2R,3R)-2-(2,4-difluorephenyl)-3 - (4- methylenepiperidino)-
`1-(1H- 1,2,4-triazol- 1-yl)butan-2-ol
`There was added 11.2 ml of 50% aqueous solution of
`55 potassium hydroxide to 1.336 g of 4-methylenepiperidine
`hydrochloride and, after dissolved under stirring, the result-
`ing solution was extracted with 20 mi of ethyl ether. Then
`the aqueous phase was fuaher extracted with 10 mi of ethyl
`ether, and the organic phases were combined and ethyl ether
`6o was removed therefrom. To the residue there were added 3
`ml of ethanol, 251 mg of (2R3S)-2-(2.4-difluorophenyl)-3-
`methyl-2-[(1H-1.2.4-triazol-l-yl)methyl]oxirane and 3 ml
`of distilled water in order, and the mixture was refluxed with
`heating for 24 hours in the oil bath at 85° C. After the
`65 reaction, the reaction solution was cooled to room
`temperature+ and thereto were added 20 ml of ethyl acetate
`and 20 ml of distilled water, and the organic phase was
`
`TABLE 4
`
`Number of died mice/number of tested mice
`
`Dose
`
`subcutaneous
`
`1000 mg/kg
`5OO m,g&g
`250 mg/kg
`125 l:ng/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-
`pound of the present invention has low toxicity.
`The compounds of the present invention have strong
`anfifungal activity and low toxicity. A fungicide containing
`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, Trichopbyton, Microsporurn, Epidermophyton,
`Malassezia, Cryptococcus neoformans, Aspergillus,
`
`Page 6
`
`

`

`5,716,969
`
`11
`separated. The aqueous phase was further extracted with 10
`ml of ethyl 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 solvent was removed. The residue was subjected to
`HPLC 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 rag
`of the rifled compound. Yield: 54.0%. Upon recrystMllzation
`from a mixed solvent of etherihexane, a pure product having
`a melting point of 860-87° C. was obtained.
`HPLC: The analysis was carried out using hexane/isopropyl
`alcohol of 9/1 as a mobile phase, at a flow rate of 1.0
`mi/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: [cx]D2s-93° (C=I.00, CHC13)
`Elemental analysis: For ClsH22F2N40 Calculated: C, 62.15;
`H, 6.36; N, 16.02 Found: C, 62.05; H, 6.37; N, 16.08
`XH-NMR spectrum (CDCI3) ~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)
`
`12
`difluorophenyl)-8-methyl-2-[(1H- 12A-triazol- 1 -yl)methyl]
`oxirane, (2S3R)-2-(2,4-difluorophenyl)-3 -methyl-2- [(1H-
`1,2,4-triazol-l-yl)methyl]oxirane being an enantiomer
`thereof was used.
`5 HPLC: The analysis was carried out using hexane/isopropyl
`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
`10 appeared at a retention time of 5.8 minutes.
`1H-NMR spectrum (CDCI3) ~3ppm: 0.96 (3H,dd, J=3 Hz,7
`Hz), 2.1-2.5 (6H,m), 2.6--2.8 (2Hart), 2.91 (1H,q.J=7 Hz),
`4.64 (2H,s), 4.80 (1H,dJ=15 Hz), 4.89 (1H,dJ=15 Hz),
`5.48 (1H,brs), 6.7--6.8 (2H,m), 7.5-7.6 (1Hart), 7.78
`15 (1H,s), 8.03 (1H,s)
`
`2O
`
`EXAMPLES 4 TO 14
`
`EXAMPLE 2
`
`The compounds of Examples 4 to 14 shown in Table 6
`25 were synthesized using starting materials shown in Table 5
`
`in the same manner as in Example 1.
`
`TABLE 5
`
`Epoxy compound (ID
`
`Amine derivative (I~)
`
`(2RS.3RS)-2-(2.4-difluorophenyl)-3-(4-
`methylenepiperidino)- 1-(1H- 1.2,4-triazol- 1-yl)butan-2-ol
`The rifled compound was obtained in the same manner as
`in Example 1 except that instead of (2R3S)-2-(2,4-
`difluorophenyl)-3-methyl-2-I(1H- 1.2.4-triazol- 1-yl)methyl] 30
`oxirane. (2RS3R)-2-(2,4-difluorophenyl)-3-methyl-2-[(1H-
`1.2.4-triazol-l-yl)methyl]oxirane being a racemic
`modification thereof was used.
`HPLC: The analysis was carried out using hexane/isopropyl
`alcohol of 9/1 as a mobile phase, at a flow rate of 1.0 35
`ml/min at room temperature under the conditions capable
`of detecting with UV (254 rim), and then two peaks
`having an area ratio thereof of 1:1 appeared at retention
`times of 6.6 minute and 5.8 minute, respectively.
`tH-NMR spectrum (CDC13) ~ppm: 0.96 (3H,dd, J=3 Hz, 7 40
`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).
`5.47 (1H,brs), 6.7-6.8 (2Ha’n), 7.5-7.6 (1Hart), 7.79
`(1H,s), 8.02 (1H,s)
`
`45
`
`EXAMPLE 3
`
`(2S,3S)-2-(2,4-difluorophenyl)-3-(4-methylenepiperidino)-
`1-(1H- 1,2,4-triazol-l-yl)butan-2-o1
`The rifled compound was obtained in the same manner as
`in Example 1 except that instead of (2R3S)-2 -(2,4-
`
`N
`
`N
`
`O
`
`II x :
`t
`,~/ \
`I
`.2
`
`CH2--C
`
`CH--CH3
`
`A.r
`
`/CH2)m
`
`\
`
`\
`
`(Cll2)n
`
`Rl
`
`/
`R~
`
`X
`
`*1 *2
`
`Ar
`
`Rt
`
`R2
`
`m
`
`n
`
`4
`5
`6
`7
`8
`9
`10
`11
`12
`13
`14
`
`CH R
`S
`CH RS SR
`N R