United States Patent [19J
`Naito et al.
`
`111111111111111111111111111111111111111111111111111111111111111111111111111
`US005962476A
`[11] Patent Number:
`[45] Date of Patent:
`
`5,962,476
`*Oct. 5, 1999
`
`[54] AZOLYLAMINE DERIVATIVE
`
`5,620,994
`
`4/1997 Naito eta!. ............................. 514/326
`
`[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
`
`[ *] Notice:
`
`This patent is subject to a terminal dis(cid:173)
`claimer.
`
`[21] Appl. No.: 08/966,527
`
`[22] Filed:
`
`Nov. 10, 1997
`
`Related U.S. Application Data
`
`[62] Division of application No. 08/781,204, Jan. 9, 1997, Pat.
`No. 5,716,969, which is a division of application No.
`08/532,800, filed as application No. PCT/JP94/00737, May
`2, 1994, Pat. No. 5,620,994.
`
`[30]
`
`Foreign Application Priority Data
`
`May 10, 1993
`
`[JP]
`
`Japan .................................... 5-132931
`
`Int. Cl. 6
`....................... A61K 31/445; C07D 401/06
`[51]
`[52] U.S. Cl. ............................................. 514/326; 546/210
`[58] Field of Search .............................. 514/326; 546/210
`
`[56]
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`4,507,484
`
`3/1985 Gymer et a!. .
`
`FOREIGN PATENT DOCUMENTS
`
`0 054 974
`6/1982 European Pat. Off ..
`2 159 148 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, as an effective
`ingredient, a compound having the general formula (I):
`
`(I)
`
`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.
`
`10 Claims, No Drawings
`
`Page 1 of 11
`
`ACRUX DDS PTY LTD. et al.
`
`EXHIBIT 1505
`
`IPR Petition for
`
`U.S. Patent No. 5,962,476
`
`

`

`5,962,476
`
`1
`AZOLYLAMINE DERIVATIVE
`
`This is a division of application Ser. No. 08/781,204,
`now U.S. Pat. No. 5,716,969 filed Jan. 9, 1997, which is a
`division of application Ser. No. 08/532,800 filed Nov. 7,
`1995, now U.S. Pat. No. 5,620,994, issued Apr. 15, 1997,
`which was a §371 national phase of international application
`PCT/JP94/00737 filed May 2, 1994, claiming priority from
`Japanese patent application Ser. No. 132,931 filed May 10,
`1993.
`
`TECHNICAL FIELD
`
`2
`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(cid:173)
`configuration or an acid addition salt thereof and other
`optical isomer.
`The present invention also provides a fungicide contain(cid:173)
`ing the above-mentioned compound having the general
`formula (I) or an acid addition salt thereof as an effective
`ingredient, and a process for treating mycosis using the
`10 above-mentioned compound.
`
`BEST MODE FOR CARRYING OUT THE
`INVENTION
`
`'lbe present invention relates to an azolylamine derivative
`which is effective for treatment for mycosis in human and 15
`animals and useful as fungicides for agricultural and horti(cid:173)
`cultural use or industrial use.
`
`BACKGROUND ART
`
`In the above-mentioned general formula (I), the substi-
`tuted phenyl group is a phenyl group having 1 to 3 substitu(cid:173)
`ents selected from a halogen atom and trifiuoromethyl, and
`includes, for instance, 2,4-difiuorophenyl, 2,4-
`dichlorophenyl, 4-fiuorophenyl, 4-chlorophenyl,
`2-chlorophenyl, 4-trifiuoromethylphenyl, 2-chloro-4-
`Azolylamine derivatives having, in the molecule, both of 20 fiuorophenyl, 4-bromophenyl or the like.
`an azolyl group such as triazolyl group or imidazolyl group
`The lower alkyl group includes, for instance, a straight
`and a cyclic amino group such as piperidino group, pyrro(cid:173)
`chain, branched chain or cyclic alkyl group having 1 to 6
`lidino group or morpholino group are described in JP-A
`carbon atoms such as methyl, ethyl, n-propyl, isopropyl,
`(Japanese Unexamined Patent Publication)-140768/1982
`n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl,
`and GB-A-2159148. However, it is hard to say in the aspect 25
`neopentyl and tert-pentyl.
`of an antifungal action etc. that each compound has suffi(cid:173)
`The non-substituted aryl group includes, for instance,
`cient efficacy as a medicament. Furthermore, any compound
`phenyl, naphthyl, biphenyl, or the like.
`having methylene group or a substituted methylene group on
`The substituted aryl group includes, for instance, 2,4-
`the cyclic amino group is not disclosed therein.
`30 difiuorophenyl, 2,4-dichlorophenyl, 4-fiuorophenyl,
`The present invention provides a novel azolylamine
`4-chlorophenyl, 2-chlorophenyl, 4-trifiuoromethylphenyl,
`derivative showing the potent antifungal activity which is
`2-chloro-4-fiuorophenyl, 4-bromophenyl, 4-tert(cid:173)
`characterized by having methylene group or a substituted
`butylphenyl, 4-ntrophenyl, or the like.
`methylene group on the cyclic amino group.
`The alkenyl group includes, for instance, vinyl,
`35 1-propenyl, styryl, or the like.
`The alkynyl group includes, for instance, ethynyl, or the
`like.
`The aralkyl group includes, for instance, benzyl,
`naphthylmethyl, 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-
`45 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
`50 having the general formula (I) includes one of these isomers
`or a mixture thereof.
`Among these, the compound wherein an absolute con(cid:173)
`figuration of the asymmetric carbon atoms is R,R(cid:173)
`configuration, has particularly potent antifungal action and
`55 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-difiuorophenyl)-3-( 4-methylenepiperidino )-
`1-(1H -1,2,4-triazol-1-yl)butane-2-ol,
`(2S,3S)-2-(2,4-difiuoropheny 1)-3-( 4-me thy lenepiperidino )-
`1-(1H -1,2,4-triazol-1-yl)butan-2-ol,
`(2RS, 3 R S)- 2 -(2, 4- difi uo rop hen yl)- 3- ( 4-
`methylenepiperidino )-1-(1H -1,2,4-triazole-1-yl)butane-
`2-ol,
`(2R,3R)-2-(2,4-difiuorophenyl)-3-( 4-methylenepiperidino )-
`1-(1H -imidazol-1-yl)butan-2-ol,
`
`DISCLOSURE OF THE INVENTION
`
`The present invention provides a compound having the
`general formula (I):
`
`(I)
`
`40
`
`wherein
`Ar is non-substituted phenyl group or a phenyl group
`substituted with 1 to 3 substituents selected from a
`halogen atom and trifiuoromethyl,
`R1 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 substituents
`selected from a halogen atom and a lower alkyl group,
`an alkenyl group, an alkynyl 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 65
`wherein an absolute configuration of the asymmetric carbon
`atoms with *1 and *2 is R,R-configuration, and the com-
`
`60
`
`Page 2 of 11
`
`

`

`5,962,476
`
`5
`
`10
`
`15
`
`3
`(2S,3S)-2-(2,4-difiuoropheny1)-3-( 4-methy1enepiperidino )-
`1-(1H -imidazo1-1-y1)butan-2-o1,
`(2RS ,3RS) -2-(2 ,4-difi uo rop heny1) -3 -( 4-
`methy1enepiperidino )-1-(1H-imidazo1-1-y1)butan-2-o1,
`(2R,3R)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-1-
`(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2S,3S)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-1-
`(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2RS,3RS)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-
`1-(1H-1,2,4-triazo1-1-y1)butan-2-o1,
`(2R,3R)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-1-
`(1H -imidazo1-1-y1)butan-2-o1,
`(2S,3S)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-1-
`(1H -imidazo1-1-y1)butan-2-o1,
`(2RS,3 RS)-2-( 4-ch1oropheny1)-3-( 4-methy1enepiperidino )-
`1-(1H-imidazo1-1-y1)butan-2-o1,
`(2R, 3 R)- 2- ( 4- trifi u oro me th y1p hen y1)- 3 -( 4-
`methy1enepiperidino )-1-(1H -1,2,4-triazo1e-1-y1)butan-2-
`o1,
`(2S, 3 S)- 2- ( 4- trifi uo rome th y1p hen y1)- 3- ( 4-
`methy1enepiperidino )-1-(1H-1,2,4-triazo1-1-y1)butan-2- 20
`o1,
`( 2RS, 3 RS)- 2- ( 4- trifi uo rome th y1p hen y 1)- 3- ( 4-
`methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)butan-2-
`o1,
`(2R,3R)-2-( 4-trifiuoromethy1pheny1)-3-( 4-
`methy1enepiperidino )-1-(1H-imidazo1-1-y1)butan-2-o1,
`(2S, 3 S)- 2- ( 4- trifi uo rome th y1p hen y1)- 3- ( 4-
`methy1enepiperidino )-1-(1H-imidazo1-1-y1)butane-2-o1,
`( 2RS, 3 RS)- 2- ( 4- trifi uo rome th y1p hen y 1)- 3- ( 4-
`methy1enepiperidino )-1-y1)butan-2-o1,
`(2R,3R )-2-(2,4-dich1oropheny 1)-3-( 4-methy 1enepiperidino )-
`1-(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2S,3S)-2-(2, 4-dich1oropheny1)-3-( 4-methy1enepiperidino )-
`1-(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2RS ,3 R S 3-2- (2, 4- dich1o rop hen y1)- 3- ( 4-
`methy1enepiperidino )-1-(1H-1,2,4-triazo1-1-y1)butan-2- 35
`o1,
`(2R,3R )-2-(2,4-dich1oropheny 1)-3-( 4-methy 1enepiperidino )-
`1H-imidazo1-i -y1)butan-2-o1,
`(2S,3S)-2-(2,4-dich1oropheny1)-3-( 4-methy1enepiperidino )-
`1-(1H -imidazo1-1-y1)butan-2-o1,
`(2RS ,3 R S)- 2- (2 ,4- dich1o rop hen y1)- 3- ( 4-
`methy1enepiperidino )-1-(1H-imidazo1-1-y1)butan-2-o1,
`(2R,3R)-2-(2,4-difiuoropheny1)-3-( 4-ethy1idenepiperidino )-
`1-(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2S,3S)-2-(2,4-difiuoropheny1)-3-( 4-ethy1idenepiperidino )- 45
`1-(1H -1,2,4-lriazol-1-y 1)bulan-2-o1,
`(2RS ,3RS) -2-(2 ,4-difi uo rop heny1) -3 -( 4-
`ethy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)butan-2-
`o1,
`(2R, 3 R) -2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`propy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2S, 3 S)- 2- (2, 4 -difi uo rop hen y1)- 3-3- ( 4-
`propy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2RS ,3RS) -2-(2 ,4-difi uo rop heny1) -3 -( 4-
`propy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4- n(cid:173)
`buty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan -2- 60
`o1,
`(2S, 3 S)- 2- (2, 4 -difi uo rop hen y1)- 3- ( 4 -n(cid:173)
`buty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan -2-
`o1,
`( 2 R S , 3 R S ) - 2 - ( 2 , 4 - d i fi u o r o p h e n y 1) - 3 - ( 4 - n -
`buty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan -2-
`o1,
`
`4
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4- n(cid:173)
`penty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2S, 3 S)- 2- (2 ,4 -d ifi u o rop hen y1)- 3 -( 4- n(cid:173)
`penty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2RS, 3 R S) -2- (2, 4- difi uo rop hen y1)- 3- ( 4- n(cid:173)
`penty1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`-2-(2,4-difiuoropheny1)-3-3-( 4-n(cid:173)
`(2R,3R)
`hexy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2S, 3 S)- 2- (2 ,4 -d ifi u o rop hen y1)- 3 -( 4- n(cid:173)
`hexy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2RS, 3 R S) -2- (2, 4- difi uo rop hen y1)- 3- ( 4- n(cid:173)
`hexy1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-2-
`o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclopropy1methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2S, 3 S)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclopropy1methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2RS, 3 R S)- 2 -(2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclopropy1methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclohexy1methy 1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2S, 3 S)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclohexy1methy 1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2RS, 3 R S)- 2 -(2, 4- difi uo rop hen y1)- 3- ( 4-
`cyclohexy1methy 1enepiperidino )-1-(1H -1,2,4-triazo1-1-
`y1)butan-2-o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`benzy 1idenepiperidino )-1-( 1H -1,2,4-triazo1-1-y 1)butan-2-
`o1,
`(2S,3S)-2-(2,4-difiuoropheny1)-3-( 4-benzy1idenepiperidin)-
`1-(1H -1,2,4-triazo1-1-y1)butan-2-o1,
`(2RS, 3 R S)- 2 -(2, 4- difi uo rop hen y1)- 3- ( 4-
`benzy 1idenepiperidino )-1-( 1H -1,2,4-triazo1-1-y 1)butan-2-
`o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`isopropy 1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)
`bulan-2-o1,
`(2S, 3 S)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`isopropy 1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)
`butan-2-o1,
`50 ( 2 R S , 3 R S) - 2- ( 2, 4- d i fi u oro ph en y 1)- 3 - ( 4-
`isopropy 1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)
`butan-2-o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`dipheny1methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)
`butan-2-o1,
`(2S, 3 S)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`dipheny1methy1enepiperidino )-1-( 1H -1,2,4triazo1-1-y1)
`butan-2-o1,
`(2RS, 3 R S)- 2 -(2, 4- difi uo rop hen y1)- 3- ( 4-
`dipheny1methy1enepiperidino )-1-(1H -1,2,4-triazo1-1-y 1)
`butan-2-o1,
`(2R, 3 R)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`propeny1idenepiperidino )-1-(1H -1,2,4-triazo1-1-y1)butan-
`2-o1,
`(2S, 3 S)- 2- (2, 4- difi uo rop hen y1)- 3- ( 4-
`propeny 1idenepiperidino )-1-(1H -1 ,2,4-triazo1-1-y 1)
`butan--2-o1,
`
`25
`
`30
`
`40
`
`55
`
`65
`
`Page 3 of 11
`
`

`

`5,962,476
`
`6
`
`r-N
`X
`)
`'N
`I
`I
`CH2-C-CH-OH
`I
`Ar
`
`OH CH
`
`I 3
`
`(IV)
`
`10
`
`wherein Ar and X have the same meanings as defined above,
`is reacted in the presence of a base with a compound having
`the formula R3 S02-0-S02R3 or R3S02-Z, wherein R3
`is a lower alkyl group, a halogenated lower alkyl group, or
`15 a phenyl group which may be substituted, and Z is a leaving
`group such as a halogen atom, to give a compound (V):
`
`(V)
`
`5
`(2RS ,3RS) -2-(2 ,4-difi uo rop henyl) -3 -( 4-
`propenylidenepiperidino )-1-(1H -1,2,4-triazol-1-yl)butan-
`2-ol,
`(2R, 3 R) -2- (2, 4- difi uo rop hen yl)- 3- ( 4-
`propynylidenepiperidino )-1-(1H -1,2,4-triazol-1-yl)butan-
`2-ol,
`(2S, 3 S)- 2- (2, 4 -difi uo rop hen y 1)- 3- ( 4-
`propynylidenepiperidino )-1-(1H -1,2,4-triazol-1-yl)butan-
`2-ol,
`(2RS ,3RS) -2-(2 ,4-difi uo rop henyl) -3 -( 4-
`propynylidenepiperidino )-1-(1H -1,2,4-triazol-1-yl)butan-
`2-ol,
`(2R,3R)-2-(2,4-difiuorophenyl)-3-(3-methylenepiperidino )-
`1-(1H -1,2,4-triazol-1-yl)butan-2-ol,
`(2S,3S)-2-(2,4-difiuorophenyl)-3-(3-methylenepiperidino )-
`1-(1H -1,2,4-triazol-1-yl)butan-2-ol,
`( 2 R S, 3 R S)- 2- ( 2, 4- d i fi u oro ph en y 1)- 3- ( 3-
`methylenepiperidino )-1-(1H-1,2,4-triazol-1-yl)butan-2- 20
`ol,
`(2R, 3 R) -2- (2, 4- difi uo rop hen yl)- 3- ( 3-
`methylenepyrrolidino )-1-(1H -1,2,4-triazol-1-yl)butan-2-
`ol,
`(2S, 3 S)- 2- (2, 4 -difi uo rop hen y 1)- 3- ( 3-
`methylenepyrrolidino )-1-(1H -1,2,4-triazol-1-yl)butan-2-
`ol,
`( 2 R S, 3 R S)- 2- ( 2, 4- d i fi u oro ph en y 1)- 3- ( 3-
`methylenepyrrolidino )-1-(1H -1,2,4-triazol-1-yl)butan-2- 30
`ol,
`
`25
`
`and the like.
`
`The compound of the present invention having the general
`formula (I) can be prepared according to the process shown 35
`as below:
`
`0 N
`
`0
`'1 1\
`I
`CH2-C-CH-CH3
`I
`'2
`Ar
`
`(II) r-N
`X
`)
`CH2-C-CH-N K
`'N
`I '1 I
`I 3
`'2
`(I)
`
`OH CH
`
`I
`Ar
`
`/(CH2)m
`
`"(CH)
`2 n
`
`R1
`
`R1
`
`, R2
`(In the above-mentioned formulae, Ar, R1
`have the same meanings as defined above.)
`
`, X, m and n
`
`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 (II) can
`be obtained according to such process as is described in
`JP-A(Japanese Unexamined Patent Publication)-191262/
`1990 etc., for example, a process wherein a compound
`having the general formula (IV):
`
`50
`
`55
`
`and then the compound (V) is reacted with a base.
`The amine derivative having the general formula (III) can
`be obtained according to the known synthetic process
`described in, for example, Chern. 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(cid:173)
`ously or in a reaction solution with an inorganic base such
`as sodium hydroxide or an organic base such as triethy(cid:173)
`lamine.
`The reaction is usually carried out using water, an organic
`40 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
`45 glycol, propylene glycol, grycerin or methyl cellosolve, an
`ether such as lelrahydrofuran, dioxane or dimelhoxyelhane,
`an amide such as N,N-dimethylformamide or N,N(cid:173)
`dimethylacetamide, 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).
`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
`60 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
`65 formula (I) can, if required, form a pharmaceutically accept(cid:173)
`able salt thereof, for example, a salt thereof with an inor(cid:173)
`ganic acid such as hydrochloric acid, sulfuric acid, nitric
`
`Page 4 of 11
`
`

`

`5,962,476
`
`7
`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 it the
`present invention having the above-mentioned general for(cid:173)
`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 10
`(MIC)
`MIC of a test compound against Candida albicans ATCC-
`10259 was determined by the both dilution method employ(cid:173)
`ing synthetic amino acid medium (SAAMF medium).
`Namely, to 3 ,ul of twofold dilution series of solution 15
`containing the test compound was added 300 ,ul of SAAMF
`medium inoculated with the fungus at the final concentration
`of 1x103 cells/ml. After thus obtained mixture was incubated
`at 35° C. for 2 days, MIC was determined by examining a
`minimum concentration of the test compound in which 20
`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 Sabouraud's agar medium. That
`is to say, a test compound was dissolved in dimethyl 25
`sulfoxide to give a solution containing the test compound in
`the concentration of 10 mg/ml. Further, thus obtained solu(cid:173)
`tion was diluted with dimethyl sulfoxide according to two(cid:173)
`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 30
`medium was added thereto, the mixture was sufficiently
`mixed to give a drug-added plate. The plate was inoculated
`with 5 ,ul of a fungus suspension at 106 cells/ml by
`Microplanter (Sakuma Seisakusho Co., Ltd.). As to
`Aspergillus fumigatus NI-5561 and Cryptococcus neofor- 35
`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
`determined by examining a minimum concentration of a test
`compound in which concentration the test compound inhib- 40
`ited the growth of the fungus. The results thereof are shown
`in Table 1. Clotrimazole and fluconazole were used as
`comparative control compounds.
`The abbreviated designation of names of the test fungi is
`as follows:
`
`45
`
`Name of fungus
`
`Candida albicans ATCC 10259
`Cryptococcus neofonnans NI-7496
`Aspergillus fumigatus NI-5561
`Trichophyton mentagrophytes KD-01
`
`Abbreviated
`designation
`
`C. a.
`Cr. n
`A.f
`T. m.
`
`The antifungal activity (the minimum inhibitory concen-
`tration MI C) of the compound of the present invention in the
`Examples against each fungus is shown in Table 1.
`
`TABLE 1
`
`Test
`compound
`
`Minimum inhibitory concentration (MIC(ug/ml)
`Test fun us
`
`(Ex. No.)
`
`C. a.
`
`2
`
`<0.025
`<0.025
`
`Cr.n.
`
`0.05
`0.1
`
`A. f.
`
`0.05
`0.1
`
`T.m.
`
`0.39
`0.39
`
`50
`
`55
`
`60
`
`65
`
`8
`
`TABLE 1-continued
`
`Test
`compound
`
`Minimum inhibitory concentration (MIC(ug/ml)
`Test fun us
`
`(Ex. No.)
`
`C. a.
`
`Cr.n.
`
`4
`5
`
`7
`
`10
`12
`13
`14
`Clotrimazole
`Fluconazole
`
`0.39
`<0.025
`<0.025
`<0.0125
`0.025
`<0.025
`<0.025
`<0.025
`0.1
`<0.025
`0.025
`0.39
`
`0.78
`<0.025
`0.025
`0.2
`0.05
`0.1
`0.025
`0.1
`0.39
`0.39
`0.2
`12.5
`
`A. f.
`
`>100
`0.05
`0.05
`6.25
`0.39
`0.2
`0.1
`0.2
`0.78
`0.39
`0.78
`>100
`
`T.m.
`
`50
`<0.025
`0.1
`3.13
`0.39
`0.78
`0.39
`0.78
`1.56
`0.78
`0.39
`>100
`
`Clotrimazole
`
`~ o-y fN
`C-Nu
`~~­l)o
`
`Fluconazole
`
`N
`
`OH
`
`fN
`
`0
`!H2 c7! CH2-N\J
`I#
`
`~F
`
`F
`
`The above-mentioned results reveal that the compound of
`the present invention having the general formula (I), espe(cid:173)
`cially the compound wherein the absolute configuration is
`R,R-configuration, has extremely high activity in compari(cid:173)
`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(cid:173)
`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/ml) was
`dropped and the skin surface was infected by rubbing it with
`a glass rod. The test compound was dissolved in polyeth(cid:173)
`ylene glycol400-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
`
`Page 5 of 11
`
`

`

`5,962,476
`
`9
`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)}x100
`
`The results are shown in Table 2. Clotrimazole was used
`as a control compound.
`
`TABLE 2
`
`Group
`
`Control (non-treated)
`Control (vehicle)
`Compound of Example 1
`Clotrimazole
`
`Inhibitory ratio
`(%)
`
`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 no of
`spore suspension of Candida albicans KC-36 (5x107 cells/
`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
`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(cid:173)
`DIDAGSAGAR 'EIKEN'(EIKENCHEMICALCO.,LTD.
`) for 7 days. The inhibitory ratio was calculated according to
`the above-mentioned formula. The results are shown in
`Table 3. Clotrimazole was employed as a control compound.
`
`TABLE 3
`
`Group
`
`Control (non-treated)
`Control (vehicle)
`Compound of Example 1
`Clotrimazole
`
`Inhibitory ratio
`(%)
`
`4
`
`98
`96
`
`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(cid:173)
`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
`
`15
`
`10
`The compounds of the present invention have strong
`antifungal 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, Trichophyton, Microsporum, Epidermophyton,
`Malassezia, Cryptococcus neoformans, Aspergillus,
`Coccidioides, Paracoccidioides, Histoplasma or Blastomy-
`10 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
`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
`20 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 external
`preparation, the preparation may be in a dosage form such
`25 as a cream, a liquid preparation, an ointment, an oculentum,
`a suppository, a vaginal suppository, a powder or an emul(cid:173)
`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.
`30 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-
`35 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
`40 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
`45 means of the following Examples and Reference Examples.
`However, it is to be understood that the present invention is
`not limited to those Examples.
`1 H-NMR spectra were determined in the solution of
`deulteriochloroform (CDC13 ) using tetramethylsilane as an
`so internal standard by means of JNM-EX270 spectrometer
`(JEOL LTD.), and a value of chemical shift (o) was
`expressed with ppm. The determination by high perfor(cid:173)
`mance liquid chromatography (hereinafter, referred to as
`HPLC) was carried out using an chiral column, CHIRAL-
`ss CEL OJ ( 4.6 mmx25 em, Daicel Chemical Industries, Ltd.)
`by means of LC-6A (HPLC apparatus, Shimadzu
`Corporation).
`
`EXAMPLE 1
`
`Dose
`
`subcutaneous
`
`1000 mg!kg
`500 mg!kg
`250 mg!kg
`125 mg!kg
`
`0/3
`0/3
`0/3
`0/3
`
`oral
`
`0/3
`0/3
`
`60
`
`(2R,3R)-2-(2,4-difiuorophenyl)-3-( 4-
`methylenepiperidino )-1-(1H -1,2,4-triazol-1-yl)
`butan-2-ol
`
`As shown in the above Table, it is found that the com(cid:173)
`pound of the present invention has low toxicity.
`
`There was added 11.2 ml of 50% aqueous solution of
`65 potassium hydroxide to 1.336 g of 4-methylenepiperidine
`hydrochloride and, after dissolved under stirring, the result(cid:173)
`ing solution was extracted with 20 ml of ethyl ether. Then
`
`Page 6 of 11
`
`

`

`5,962,476
`
`11
`the aqueous phase was further extracted with 10 ml of ethyl
`ether, and the organic phases were combined and ethyl ether
`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-triazol-1-yl)methyl]oxirane and 3 ml 5
`of distilled water in order, and the mixture was refluxed with
`heating for 24 hours in the oil bath at 85° C. After the
`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 10
`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 15
`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 mg
`of the titled compound. Yield: 54.0%. Upon recrystallization
`from a mixed solvent of ether/hexane, a pure product having 20
`a melting point of 86°-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 ml/min at room temperature under the
`conditions capable of detecting with UV (254 nm), and 25
`then a single peak appeared at a retention time of 6.6
`minutes.
`Specific rotation: [ab28-93° (C=1.99, CHC13 )
`Elemental analysis: For C18H 22F 2 N 4 0 Calculated: 30
`C,62.15; H,6.36; N,16.02 Found: C,62.05; H,6.37;
`N,16.08
`1H-NMR spectrum (CDC13 ) o ppm: 0.96 (3H,dd), 2.1-2.5
`(6H,m), 2.6-2.8 (2H,m), 2.91 (lH,q), 4.64 (2H,s), 4.80
`(lH,d), 4.89 (lH,d), 5.48 (lH,brs), 6.7-6.8 (2H,m), 35
`7.47-7.63 (lH,m), 7.79 (lH,s), 8.03 (lH,s)
`
`EXAMPLE 2
`
`(2RS,3RS)-2-(2,4-difluorophenyl)-3-( 4-
`methylenepiperidino )-1-(1H -1,2,4-triazol-1-yl)
`butan-2-ol
`
`40
`
`12
`oxirane, (2RS,3SR)-2-(2,4-difluorophenyl)-3-methyl-2-
`[(1H-1,2,4-triazol-1-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 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 minute and 5.8
`minute, respectively.
`1 H-NMR spectrum (CDC13 ) o ppm: 0.96 (3H,dd,J=3 Hz,7
`Hz), 2.1-2.5 (6H,m), 2.6-2.8 (2H,m), 2.91 (lH,q,J=
`7Hz), 4.64 (2H,s), 4.80 (1H,d,J=15Hz), 4.89 (lH,d,J=
`15Hz), 5.47 (lH,brs), 6.7-6.8 (2H,m), 7.5-7.6 (lH,m),
`7.79 (lH,s), 8.02 (lH,s)
`
`EXAMPLE 3
`
`(2S,3S)-2-(2,4-difluorophenyl)-3-( 4-
`methylenepiperidino )-1-(1H-l, 2,4-triazol-1-yl)
`butan-2-ol
`
`The titled compound was obtained in the same manner as
`in Example 1 except that instead of (2R,3S)-2-(2,4-
`difluorophenyl)-3-methyl-2-[ (1H -1,2,4-triazol-1-yl)methyl]
`oxirane, (2S,3R)-2-(2,4-difluorophenyl)-3-methyl-2-[(1H-
`1,2,4-triazol-1-yl)methyl]oxirane being an enantiomer
`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 ml/min at room temperature under the conditions
`capable of detecting with UV (254 nm), and then a
`single peak appeared at a ret