AU2001234095B2 - Cyclic hexapeptide derivatives - Google Patents

Cyclic hexapeptide derivatives Download PDF

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AU2001234095B2
AU2001234095B2 AU2001234095A AU2001234095A AU2001234095B2 AU 2001234095 B2 AU2001234095 B2 AU 2001234095B2 AU 2001234095 A AU2001234095 A AU 2001234095A AU 2001234095 A AU2001234095 A AU 2001234095A AU 2001234095 B2 AU2001234095 B2 AU 2001234095B2
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Prior art keywords
substituted
phenyl
compound
alkyl
alkoxy
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AU2001234095A1 (en
Inventor
David Barrett
Hiroshi Matsuda
Keiji Matsuda
Takahiro Matsuya
Hiroaki Mizuno
Kenji Murano
Takashi Ogino
Ayako Toda
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Astellas Pharma Inc
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Astellas Pharma Inc
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Priority claimed from AUPQ5752A external-priority patent/AUPQ575200A0/en
Priority claimed from AUPQ9552A external-priority patent/AUPQ955200A0/en
Priority claimed from AUPR2344A external-priority patent/AUPR234400A0/en
Application filed by Astellas Pharma Inc filed Critical Astellas Pharma Inc
Priority to AU2001234095A priority Critical patent/AU2001234095B2/en
Priority claimed from PCT/JP2001/001204 external-priority patent/WO2001060846A1/en
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Description

WO 01/60846 PCT/JP01/01204
DESCRIPTION
CYCLIC HEXAPEPTIDE DERIVATIVES TECHNICAL FIELD The present invention relates to new polypeptide compounds and salts thereof which are useful as a medicament.
BACKGROUND ART In U.S. Pat. No. 5,376,634, 5,569,646, WO 96/11210 and WO 99/40108, there are disclosed the polypeptide compound and a pharmaceutically acceptable salt thereof, which have antimicrobial activities (especially antifungal activity).
DISCLOSURE OF INVENTION The present invention relates to new polypeptide compound and a salt thereof.
More particularly, it relates to new polypeptide compound and a salt thereof, which have antimicrobial activities [especially, antifungal activities, in which the fungi may include Aspergillus, Cryptococcus, Candida, Mucor, Actinomyces, Histoplasma, Dermatophyte, Malassezia, Fusarium and the like.], inhibitory activity on P-1,3-glucan synthase, and further which are expected to be useful for the prophylactic and/or therapeutic treatment of Pneumocystis carinii infection Pneumocystis carinii pneumonia) in a human being or an animal, to a process for preparation thereof, to a pharmaceutical composition comprising the same, and to a method for the prophylactic and/or therapeutic treatment of infectious disease including Pneumocystis carinii infection Pneumocystis carinii pneumonia) in a human being or an animal.
The object polypeptide compounds of the present invention are new and can be represented by the following general formula WO 01/60846 PCT/JP01/01204 2
OH
OH O
NH-R
0 R2 HO 0 H OH \N H (I) R3/ H 0 CH 3 NH OH 0 OH
R
6 wherein
R
1 is hydrogen or acyl group,
R
2 is hydrogen or acyl group,
R
3 is lower alkyl which has one or more hydroxy or protected hydroxy, R4 is hydrogen or hydroxy, is hydrogen, hydroxy, lower alkoxy or hydroxysulfonyloxy, and
R
6 is hydroxy or acyloxy, or a salt thereof.
The new polypeptide compound or a salt thereof can be prepared by the process as illustrated in the following reaction schemes.
WO 01/60846 PCT/JP01/01204 Process 1
R
3 =0 (III) or its reactive derivative or a salt thereof Rb
(II)
or its reactive derivative at the amino group or a salt thereof or a salt thereof WO 01/60846 PCT/JP01/01204 Process 2
R
2 -OH (IV) a or its reactive derivative at the carboxy group or a salt thereof or its reactive derivative at the amino group or a salt thereof R2 a*--IN (Ib) or a salt thereof WO 01160846 WO 0160846PCT/JPOI/01204 Process 3 elimination reaction
R
6 (1b) or a salt thereof or a salt thereof WO 01160846 WO 0160846PCT/IPOI/01204 Process 4 Rl-OH a or its reactive derivative at the carboxy group or a salt thereof
R,
6 (To) or its reactive derivative at the amino group or a salt thereof (ITd) or a salt thereof WO 01/60846 PCT/JP01/01204 7 wherein R 1
R
2
R
3
R
4
R
5 and R 6 are defined above, Ra is acyl group, and Ra is acyl group.
Suitable salt of the new polypeptide compound is a pharmaceutically acceptable and conventional non-toxic salt, and may include a salt with a base or an acid addition.salt such as a salt with an inorganic base, for example, an alkali metal salt sodium salt, potassium salt, etc.), an alkaline earth metal salt calcium salt, magnesium salt, etc.), an ammonium salt; a salt with an organic base, for example, an organic amine salt triethylamine salt, diisopropylethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, 4-dimethylaminopyridine salt, etc.); an inorganic acid addition salt hydrochloride hydrobromide, sulfate, phosphate, etc.); an organic carboxylic sulfonic acid addition salt formate, acetate, trifluoroacetate, maleate, tartrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.); a salt with a basic or acidic amino acid arginine, aspartic acid, glutamic acid, etc.).
Suitable examples and illustration of the various definitions in the above and subsequent descriptions of the present specification, which the present invention intends to include within the scope thereof, are explained in detail as follows: The term "lower" is used to intend a group having 1 to 6 carbon atom(s), unless otherwise provided.
Suitable example of "one or more" may be the number of 1 to 6, in which the preferred one may be the number of 1 to 3, and WO 01/60846 PCT/JP01/01204 8 the most preferred one may be the number of 1 or 2.
Suitable example of "halogen" may be fluorine, chlorine, bromine, iodine and the like.
Suitable example of "lower alkoxy" may include straight or branched one such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy, tert-pentyloxy, neopentyloxy, hexyloxy, isohexyloxy and the like.
Suitable example of "higher alkoxy" may include straight or branched one such as heptyloxy, octyloxy, 3,5-dimethyloctyloxy, 3,7-dimethyloctyloxy, nonyloxy, decyloxy, undecyloxy, dodecyloxy, tridecyloxy, tetradecyloxy, hexadecyloxy, heptadecyloxy, octadecyloxy, nonadecyloxy, icosyloxy, and the like.
Suitable example of "lower alkyl" may include straight or branched one having 1 to 6 carbon atom(s), such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neo-pentyl, hexyl, isohexyl and the like.
Suitable example of "higher alkyl" may include straight or branched one such as heptyl, octyl, 3,5-dimethyloctyl, 3,7dimethyloctyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, and the like.
Suitable example of "aryl" and "ar" moiety may include phenyl which may have lower alkyl phenyl, mesityl, xylyl, tolyl, etc.), naphthyl, anthryl, indanyl, fluorenyl, and the like, and this "aryl" and "ar" moiety may have one or more halogen.
Suitable example of "aroyl" may include benzoyl, toluoyl, naphthoyl, anthrylcarbonyl, and the like.
Suitable example of "heterocyclic group" may include unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, dihydropyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl 4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, etc.), tetrazolyl (e.g.
WO 01/60846 PCT/JP01/01204 9 1H-tetrazolyl, 2H-tetrazolyl, etc.), etc.; saturated 3 to 8-membered (more preferably 5 or 6-membered) heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, pyrrolidinyl, imidazolidinyl, piperidyl, piperazinyl, azetidinyl, etc.; unsaturated condensed heterocyclic group containing 1 to 4 nitrogen atom(s), for example, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, oxazolyl, isoxazolyl, oxadiazolyl 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.), etc.; saturated 3 to 8-membered (more preferably 5 or 6-membered) heteromonocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, morpholinyl, sydnonyl, morpholino, etc.; unsaturated condensed heterocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, benzoxazolyl, benzoxadiazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example, thiazolyl, isothiazolyl, thiadiazolyl 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.), dihydrothiazinyl, etc.; saturated 3 to 8-membered (more preferably 5 or 6-membered) heteromonocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example thiazolidinyl, thiomorpholinyl, thiomorpholino, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 sulfur atom(s), for example, thienyl, dihydrodithiinyl, dihydrodithionyl, etc.; unsaturated condensed heterocyclic group containing 1 or 2 WO 01/60846 PCT/JP01/01204 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example, benzothiazolyl, benzothiadiazolyl, imidazothiadiazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing an oxygen atom, for example, furyl etc.; saturated 3 to 8-membered (more preferably 5 or 6-membered) heteromonocyclic group containing 1 or 2 oxygen atom(s), for example, tetrahydrofuran, tetrahydropyran, dioxacyclopentane, dioxacyclohexane, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing an oxygen atom and 1 or 2 sulfur atom(s), for example, dihydrooxathiinyl, etc.; unsaturated condensed heterocyclic group containing 1 or 2 sulfur atom(s), for example benzothienyl, benzodithiinyl, etc.; unsaturated condensed heterocyclic group containing an oxygen atom and 1 or 2 sulfur atom(s), for example, benzoxathiinyl, etc.; and the like, and this "heterocyclic group" may have one or more suitable substituent(s) selected from the group consisting of lower alkyl, oxo, cyclo(lower)alkyl, hydroxy(lower)alkyl, carboxy(lower)alkanoyl which may have amino and heterocycliccarbonyl.
Suitable example of "cyclo(lower)alkyl" may include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like, and this "cyclo(lower)alkyl" may have one or more lower alkyl.
Suitable example of "cyclo(lower)alkyloxy" may include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.
Suitable example of "acyl group" may include aliphatic acyl, aromatic acyl, arylaliphatic acyl and heterocyclic-aliphatic acyl derived from carboxylic acid, carbonic acid, carbamic acid, sulfonic acid, and the like.
Suitable example of said "acyl group" may be illustrated as follows.
WO 01160846 PCT/JPOI/01204 Carboxy; carbarnoyl; mono or di(lower)alkylcarbamoyl rethylcarbamoyl, dimethylcarbamoy-, ethylcarbamoyl, diethylcarbamoyl, etc.) Aliphatic acyl such as lower or higher alkanoyl formyl, acetyl, propanoyl, butanoyl, 2-methyipropanoyl, pentanoyl, 2, 2-dimethyipropanoyl, hexanoyl, heptanoyl, octanoy-, nonanoyl, decanoyl, undecanoyl, clocecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, icosanoyl, etc.); lower or higher alkoxycarbonyl methoxycarbolyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbor'yl, heptyloxycarbonyl, etc.); lower alkenyloxycarbonyl vinyloxycarbonyl, propenyloxycarbonyl, allyloxycarbonyl, butenyloxycarbonyl, but edienyl oxyca rbolyl, pent en yloxyca rbonyl, hexenyloxycarbonly, etc.); lower or higher alkylsulfolyl rethylsulfonyl, ethylsulfonyl, etc.); lower or higher alkoxysulfonyl methoxysulfonyl, ethoxysulfonyl, etc.); or the like; Aromatic acyl such as aroyl benzoyl, toluoyl, naphthoyl, etc.); ar(lower)alkanoyl phenyl(CI-C 6 )alkaloyl phenylacetyl, phenyipropanoyl, phenylbutafloyl, phenylisobutanoyl, phenylpentanoyl, phenyih-exanoyl, etc.), naphthyl (Cl-C 6 )alkanoyl naphthylacetyl, naphthylpropanoyl, naphthylbutaioyl, etc.), etc.]; ar(lower)alkenoyl phenyl(C 3
-C
6 )alkenoyl phenyipropenoyl, phenylbutenoyl, phenyirnethacryloyl, phenylpentanoyl, phenylhexenoyl, etc.), naphthyl (0 3
-C
6 )alkenoyl naphthylpropenoyl, naphthylbutenoyl, etc.), etc.]; ar(lower)alkoxycarboflyl phenyl (Cl-C 6 )alkoxycarbonyl benzyloxycarboflyl, etc.), fluorenyl (Cl-C 6 )alkoxycarbonyl fluorenylmethyloxYcrbolyl, etc.), etc.]; WO 01/60846 PCT/JP01/01204 12 aryloxycarbonyl phenoxycarbonyl, naphthyloxycarbonyl, etc.); aryloxy(lower)alkanoyl phenoxyacetyl, phenoxypropionyl, etc.); arylcarbamoyl phenylcarbamoyl, etc.); arylthiocarbamoyl phenylthiocarbamoyl, etc.); arylglyoxyloyl phenylglyoxyloyl, naphthylglyoxyloyl, etc.); arylsulfonyl which may have 1 to 4 lower alkyl phenylsulfonyl, p-tolylsulfonyl, etc.); aroyl benzoyl) substituted with one or more suitable substituent(s); or the like; Heterocyclic acyl such as heterocycliccarbonyl; heterocyclic(lower)alkanoyl heterocyclicacetyl, heterocyclicpropanoyl, heterocyclicbutanoyl, heterocyclicpentanoyl, heterocyclichexanoyl, etc.); heterocyclic(lower)alkenoyl heterocyclicpropenoyl, heterocyclicbutenoyl, heterocyclicpentenoyl, heterocyclichexenoyl, etc.); heterocyclicglyoxyloyl; or the like; in which suitable "heterocyclic" moiety in the terms "heterocycliccarbonyl", "heterocyclic(lower)alkanoyl", "heterocyclic(lower)alkenoyl" and "heterocyclicglyoxyloyl" can be referred to aforementioned "heterocyclic" moiety.
Suitable example of "acyl group" of R 1 can be referred to aforementioned "acyl group", in which the preferred one may be lower alkoxycarbonyl, higher alkanoyl and benzoyl substituted with one or more suitable substituent(s).
Suitable example of "suitable substituent(s)" in the term of "benzoyl substituted with one or more suitable substituent may be thiadiazolyl substituted with phenyl having phenyl substituted with morphlino having lower alkyl, WO 01/60846 PCT/JP01/01204 13 thiadiazolyl substituted with phenyl having a suitable substituent selected from the group consisting of lower alkoxy(lower)alkoxy and lower alkoxy(higher)alkoxy, piperazinyl substituted with phenyl having piperidyl substituted with a suitable substituent selected from the group consisting of phenyl having lower alkoxy(lower)alkoxy, cyclo(lower)alkyloxy and lower alkoxy(lower)alkylthio, piperazinyl substituted with phenyl having phenyl substituted with morpholino having lower alkyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with a suitable substituent selected from the group consisting of lower alkoxy(lower)alkoxy and lower alkoxy(lower)alkylthio, imidazothiadiazolyl substituted with phenyl having lower alkoxy(lower)alkoxy, phenyl subsutituted with piperazinyl having phenyl substituted with morpholino having lower alkyl, isoxazolyl substituted with phenyl having lower alkoxy(lower)alkoxy, isoxazolyl substituted with phenyl having higher alkoxy substituted with morpholino having lower alkyl, thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo (lower) alkyl which has one or more suitable substituent(s) selected from the group consisting of lower alkyl, lower alkenyl, lower alkoxy(higher)alkoxy and phenyl, thiadiazolyl substituted with phenyl having piperazinyl substituted with lower alkyl having cyclo(lower)alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with one or more suitable substituent(s) selected from the group consisting of cyclo(lower)alkyloxy, lower alkoxy(lower)alkoxy and lower alkoxy(lower)alkoxy(lower)alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower)alkyl and lower alkoxy, thiadiazolyl substituted with pyridyl having piperazinyl substituted with cyclo(lower)alkyl having lower alkyl, WO 01/60846 PCT/JP01/01204 14 imidazothiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower) alkyl, imidazothiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo(lower)alkyl having lower alkyl, phenyl substituted with piperazinyl having cyclo(lower)alkyl substituted with one or more suitable substituent(s) selected from the group consisting of cyclo(lower)alkyl which may have lower alkoxy, lower alkyl, lower alkoxy and phenyl which may have lower alkoxy, in which the preferred one may be thiadiazolyl substituted with phenyl having phenyl substituted with morpholino having dimethyl, thiadiazolyl substituted with phenyl having a substituent selected from the group consisting of methoxyhexyloxy and methoxyheptyloxy, piperazinyl substituted with phenyl having piperidyl substituted with a substituent selected from the group consisting of phenyl having methoxybutoxy, cyclohexyloxy and methoxyhexylthio, piperazinyl substituted with phenyl having phenyl substituted with morpholino having dimethyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with a substituent selected from the group consisting of methoxypropoxy, methoxybutoxy, methoxypentyloxy and methoxyhexylthio, imidazothiadiazolyl substituted with phenyl having methoxybutoxy, phenyl subsutituted with piperazinyl having phenyl substituted with morpholino having dimethyl, isoxazolyl substituted with phenyl having methoxyhexyloxy, isoxazolyl substituted with phenyl having heptyloxy substituted with morphlino having dimethyl, WO 01/60846 PCT/JP01/01204 thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclohexyl which has one or two substituent(s) selected from the group consisting of methyl, methylene, methoxyheptyloxy, methoxyoctyloxy and phenyl, thiadiazolyl substituted with phenyl having piperazinyl substituted with methyl which has a substituent selected from the group consisting of cyclopentyl and cyclohexyl, thiadiazolyl substituted with phenyl having piperidyl substituted with one or two substituent(s) selected from the group consisting of cyclohexyl, methoxy, cyclohexyloxy, methoxypentyloxy, methoxyhexyloxy, methoxybutoxymethyl and methoxypentyloxymethyl, thiadiazolyl substituted with pyridyl having piperazinyl substituted with cyclohexyl which has a substituent selected from the group consisting of methyl and ethyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with cyclohexyl, imidazothiadiazolyl substituted with phenyl having piperazinyl substituted with cyclohexyl having methyl, phenyl substituted with piperazinyl having cyclohexyl substituted with one or two substituent(s) selected from the group consisting of ethyl, t-butyl, methoxy, cyclopentyl, cyclohexyl which may have methoxy or dimethyl, and phenyl which may have methoxy.
The more suitable example of "acyl group" may be benzoyl which has thiadiazolyl substituted with phenyl having phenyl substituted with morpholino having dimethyl, benzoyl which has thiadiazolyl substituted with phenyl having a substituent selected from the group consisting of methoxyhexyloxy and methoxyheptyloxy,benzoyl which has piperazinyl substituted with phenyl having piperidyl substituted with a substituent selected from the group consisting of phenyl having methoxybutoxy, cyclohexyloxy and methoxyhexylthio, WO 01/60846 PCT/JP01/01204 16 benzoyl which has piperazinyl substituted with phenyl having phenyl substituted with morpholino having dimethyl, benzoyl which has imidazothiadiazolyl substituted with phenyl having piperidyl substituted with a substituent selected from the group consisting of methoxypropoxy, methoxybutoxy, methoxypentyloxy and methoxyhexylthio, benzoyl which has imidazothiadiazolyl substituted with phenyl having methoxybutoxy, benzoyl which has phenyl subsutituted with piperazinyl having phenyl substituted with morpholino having dimethyl, benzoyl which has isoxazolyl substituted with phenyl having methoxyhexyloxy, benzoyl which has isoxazolyl substituted with phenyl having heptyloxy substituted with morphlino having dimethyl, benzoyl which has thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclohexyl which has one or two substituent(s) selected from the group consisting of methyl, methylene, methoxyheptyloxy, methoxyoctyloxy and phenyl, benzoyl which has thiadiazolyl substituted with phenyl having piperazinyl substituted with methyl which has a substituent selected from the group consisting of cyclopentyl and cyclohexyl, benzoyl which has thiadiazolyl substituted with phenyl having piperidyl substituted with one or two substituent(s) selected from the group consisting of cyclohexyl, methoxy, cyclohexyloxy, methoxypentyloxy, methoxyhexyloxy, methoxybutoxymethyl and methoxypentyloxymethyl, benzoyl which has thiadiazolyl substituted with pyridyl having piperazinyl substituted with cyclohexyl which has a substituent selected from the group consisting of methyl and ethyl, benzoyl which has imidazothiadiazolyl substituted with phenyl having piperidyl substituted with cyclohexyl, benzoyl which has imidazothiadiazolyl substituted with phenyl having piperazinyl substituted with cyclohexyl having WO 01/60846 PCT/JP01/01204 17 methyl, benzoyl which has phenyl substituted with piperazinyl having cyclohexyl substituted with one or two substituent(s) selected from the group consisting of ethyl, t-butyl, methoxy, cyclopentyl, cyclohexyl which may have methoxy or dimethyl, and phenyl which may have methoxy.
Suitable example of "lower alkyl" in the term of "lower alkyl which has one or more hydroxy or protected hydroxy" can be referred to aforementioned "lower alkyl", in which the preferred one may be methyl, ethyl, propyl, isopropyl, butyl, pentyl and hexyl.
Suitable example of "hydroxy protective group" in the term of "protected hydroxy" may include acyl lower alkanoyl, etc.) as mentioned above, phenyl(lower)alkyl which may have one or more suitable substituent(s) benzyl, 4methoxybenzyl, trityl, etc.), tri-substituted silyl tri(lower)alkylsilyl(e.g., trimethylsilyl, tbutyldimethylsilyl, etc.), etc.], tetrahydropyranyl and the like.
Suitable example of "lower alkyl which has one or more hydroxy or protected hydroxy" may be dihydroxypropyl, dihydroxyisopropyl, trihydroxybutyl, tetrahydroxypentyl, pentahydroxyhexyl and diacetyloxyisopropyl.
Suitable example of "acyl group" of R 2 can be referred to aforementioned "acyl group", in which the preferred one may be "amino protective group" mentioned below, and the most preferred one may be acetyl, 2-acetyloxypropionyl, methylsulfonyl, diaminopentanoyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl and tert-butoxycarbonyl.
Suitable example of "amino protective group" may be included in aforementioned "acyl group", a conventional protective group such as ar(lower)alkoxycarbonyl and lower alkoxycarbonyl, in WO 01/60846 PCT/JP01/01204 18 which the preferred one may be phenyl-
(C
1
-C
4 )alkoxycarbonyl and fluorenyl (C 1
-C
4 alkoxycarbonyl and (C 1
-C
4 )alkoxycarbonyl, and the most preferred one may be benzyloxycarbonyl, fluorenylmethoxycarbonyl and tertbutoxycarbonyl.
Suitable example of "acyl" moiety of "acyloxy" can be referred to aforementioned "acyl group", in which the preferred one may be lower alkenyloxycarbonyl, and the most preferred one may be allyloxycarbonyl.
Suitable example of "acyloxy" may be lower alkenyloxycarbonyloxy, and the more preferred one may be allyloxycarbonyloxy.
Particularly, the preferred examples of the cyclic polypeptide compound of the present invention are as follows: the compound wherein
R
1 is hydrogen, lower alkoxycarbonyl, higher alkanoyl or benzoyl substituted with one or more suitable substituent(s),
R
2 is hydrogen,
R
3 is lower alkyl which has one or more hydroxy,
R
4 is hydrogen or hydroxy;
R
5 is hydroxy or hydroxysulfonyloxy; and
R
6 is hydroxy.
And, more preferred one may be the compound (I) wherein
R
1 is hydrogen, lower alkoxycarbonyl, higher alkanoyl or benzoyl substituted with one or more suitable substituent(s),
R
2 is hydrogen,
R
3 is lower alkyl which has two hydroxy,
R
4 is hydrogen or hydroxy; WO 01/60846 PCT/JP01/01204 19
R
5 is hydroxy or hydroxysulfonyloxy; and
R
6 is hydroxy.
And, still more preferred one may be the compound (I) wherein
R
1 is benzoyl substituted with a suitable substituent selected from the group consisting of thiadiazolyl substituted with phenyl having phenyl substituted with morphlino having lower alkyl, thiadiazolyl substituted with phenyl having a suitable substituent selected from the group consisting of lower alkoxy(lower)alkoxy and lower alkoxy(higher)alkoxy, piperazinyl substituted with phenyl having piperidyl substituted with a suitable substituent selected from the group consisting of phenyl having lower alkoxy(lower)alkoxy, cyclo(lower)alkyloxy and lower alkoxy(lower)alkylthio, piperazinyl substituted with phenyl having phenyl substituted with morpholino having lower alkyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with a suitable substituent selected from the group consisting of lower alkoxy(lower)alkoxy and lower alkoxy(lower)alkylthio, imidazothiadiazolyl substituted with phenyl having lower alkoxy(lower)alkoxy, phenyl subsutituted with piperazinyl having phenyl substituted with morpholino having lower alkyl, isoxazolyl substituted with phenyl having lower alkoxy(lower)alkoxy, isoxazolyl substituted with phenyl having higher alkoxy substituted with morpholino having lower alkyl, thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo (lower) alkyl which has one or more suitable substituent selected from the group consisting of lower alkyl, lower alkenyl, lower alkoxy(higher)alkoxy and phenyl, thiadiazolyl substituted with phenyl having piperazinyl WO 01/60846 PCT/JP01/01204 substituted with lower alkyl having cyclo(lower)alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with one or more suitable substituent(s) selected from the group consisting of cyclo(lower)alkyloxy, lower alkoxy(lower)alkoxy and lower alkoxy(lower)alkoxy(lower)alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower)alkyl and lower alkoxy, thiadiazolyl substituted with pyridyl having piperazinyl substituted with cyclo(lower)alkyl having lower alkyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower) alkyl, imidazothiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo(lower)alkyl having lower alkyl, and phenyl substituted with piperazinyl having cyclo(lower)alkyl substituted with one or more suitable substituent(s) selected from the group consisting of cyclo(lower)alkyl which may have lower alkoxy, lower alkyl, lower alkoxy and phenyl which may have lower alkoxy,
R
2 is hydrogen,
R
3 is lower-alkyl which has two hydroxy,
R
4 is hydrogen or hydroxy;
R
5 is hydroxy or hydroxysulfonyloxy; and
R
6 is hydroxy.
And, the most preferred one may be the compound (I) wherein
R
1 is benzoyl which has thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo(lower)alkyl which has lower alkyl, benzoyl which has thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower)alkyloxy, benzoyl which has phenyl substituted with piperazinyl having cyclo(lower)alkyl substituted with WO 01/60846 PCT/JP01/01204 21 cyclo(lower)alkyl and lower alkoxy, benzoyl which has thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo(lower)alkyl,
R
2 is hydrogen,
R
3 is lower alkyl which has two hydroxy, R4 is hydrogen or hydroxy;
R
5 is hydroxy or hydroxysulfonyloxy; and
R
6 is hydroxy.
The processes for preparing the polypeptide compound of the present invention are explained in detail in the following.
Process 1 The object compound (Ia) or a salt thereof can be prepared by reacting the compound (II) or its reactive derivative at the amino group or a salt thereof with the compound (III) of the formula:
(III)
or its reactive derivative, or a salt thereof.
Suitable reactive derivative of the compound (III) may include an acid halide, an acid anhydride, an activated ester, and the like. The suitable example may be an acid chloride; acid azide; a mixed acid anhydride with an acid such as substituted phosphoric acid dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, etc.), dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, alkanesulfonic acid methanesulfonic acid, ethanesulfonic acid, etc.), sulfuric acid, alkylcarbonic acid, aliphatic carboxylic acid pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, WO 01/60846 PCT/JP01/01204 22 trichloroacetic acid, etc.); aromatic carboxylic acid benzoic acid, etc.); a symmetrical acid anydride; an activated amide with imidazole, 4-substitutd imidazole, dimethylpyrazole, triazole or tetrazole; an activated ester cyanomethyl, ester methoxymethyl ester, dimethyliminomethyl [(CH 3 2
N=CH-]
ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenylthioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, etc.); an ester with a N-hydroxy compound N,N-dimethylhydroxylamine, l-hydroxy-2-(1H)-pyridone, N-hydroxysuccinimide, N-hydroxybenzotriazole, N-hydroxyphthalimide, l-hydroxy-6-chloro-lH-benzotriazole, etc.); and the like. These reactive derivatives can optionally be selected from them according to the kind of the compound (III) to be used.
The reaction is usually carried out in a conventional solvent such as water, acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or any other organic solvent which do not adversely affect the reaction, or the mixture thereof.
When the compound (III) is used in free acid form or its salt form in the reaction, the reaction is preferably carried out in the presence of a conventional condensing agent such as N,N'dicyclohexylcarbodiimide; N-cyclohexyl-N'morpholinoethylcarbodiimide); N-cyclohexyl-N'-(4diethylaminocyclohexyl)carbodiimide; N,N'-diisopropylcarboxiimide; N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide; N,N-carbonyl-bis(2-methylimidazole); pentamethyleneketene-Ncyclohexylimine; diphenylketene-N-cyclohexylimine, ethoxyacetylene; 1-alkoxy-l-chloroethylene; trialkyl phosphite; isopropyl polyphosphate; phosphorous WO 01/60846 PCT/JP01/01204 23 oxychloride (phosphoryl chloride); phosphorous trichloride; thionyl chloride; oxalyl chloride; triphenylphosphite; 2-ethyl-7-hydroxybenzisoxazolium salt; sulfophenyl)isoxazolium hydroxide intra-molecular salt; 1-(p-chlorobenzenesulfonyloxy)-6-chloro-lH-benzotriazole; so-called Vilsmeier reagent prepared by the reaction of N,N-dimethylformamide with thionyl chloride, phosgene, phosphorous oxychloride, etc.; or the like.
The reaction may also be carried out in the presence of an organic or inorganic base such as an alkali metal bicarbonate, tri(lower)alkylamine triethylamine, diisopropylethylamine, etc.), pyridine, di(lower)alkylaminopyridine 4-dimethylaminopyridine, etc.) N-(lower)alkylmorphorine, N,N-di(lower)alkylbenzylamine, or the like.
The reaction temperature is not critical, and the reaction is usually carried out under cooling to heating.
Process 2 The object compound (Ib) or a salt thereof can be prepared by reacting the compound (Ia) or its reactive derivative at the amino group or a salt thereof with the compound (IV) of the formula: R OH (IV) (wherein R2 is acyl group) or its reactive derivative at the carboxy group or a salt thereof.
Suitable reactive derivative of the compound (IV) may include an acid halide, an acid anhydride, an activated ester, and the like. The suitable example may be an acid chloride; acid azide; a mixed acid anhydride with an acid such as substituted phosphoric acid dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, WO 01/60846 PCT/JP01/01204 24 halogenated phosphoric acid, etc.), dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, alkanesulfonic acid methanesulfonic acid, ethanesulfonic acid, etc.), sulfuric acid, alkylcarbonic acid, aliphatic carboxylic acid pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.); aromatic carboxylic acid benzoic acid, etc.); a symmetrical acid anydride; an activated amide with imidazole, 4-substitutd imidazole, dimethylpyrazole, triazole or tetrazole; an activated ester cyanomethyl, ester methoxymethyl ester, dimethyliminomethyl [(CH 3 2
N=CH-]
ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenylthioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, etc.); an ester with a N-hydroxy compound N,N-dimethylhydroxylamine, l-hydroxy-2-(1H)-pyridone, N-hydroxysuccinimide, N-hydroxybenzotriazole, N-hydroxyphthalimide, l-hydroxy-6-chloro-lH-benzotriazole, etc.); and the like. These reactive derivatives can optionally be selected from them according to the kind of the compound (IV) to be used.
The reaction is usually carried out in a conventional solvent such as water, acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or any other organic solvent which do not adversely affect the reaction, or the mixture thereof.
When the compound (IV) is used in free acid form or its salt form in the reaction, the reaction is preferably carried out in the presence of a conventional condensing agent such as N,N'dicyclohexylcarbodiimide; N-cyclohexyl-N'morpholinoethylcarbodiimide); N-cyclohexyl-N'-(4diethylaminocyclohexyl)carbodiimide; N,N'-diisopropylcarboxi- WO 01/60846 PCT/JP01/01204 imide; N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide; N,N-carbonyl-bis(2-methylimidazole); pentamethyleneketene-Ncyclohexylimine; diphenylketene-N-cyclohexylimine, ethoxyacetylene; l-alkoxy-l-chloroethylene; trialkyl phosphite; isopropyl polyphosphate; phosphorous oxychloride (phosphoryl chloride); phosphorous trichloride; thionyl chloride; oxalyl chloride; triphenylphosphite; 2-ethyl-7-hydroxybenzisoxazolium salt; sulfophenyl)isoxazolium hydroxide intra-molecular salt; 1-(p-chlorobenzenesulfonyloxy)-6-chloro-lH-benzotriazole; so-called Vilsmeier reagent prepared by the reaction of N,N-dimethylformamide with thionyl chloride, phosgene, phosphorous oxychloride, etc.; or the like.
The reaction may also be carried out in the presence of an organic or inorganic base such as an alkali metal bicarbonate, tri(lower)alkylamine triethylamine, diisopropylethylamine, etc.), pyridine, di(lower)alkylaminopyridine 4-dimethylaminopyridine, etc.) N-(lower)alkylmorphorine, N,N-di(lower)alkylbenzylamine, or the like.
The reaction temperature is not critical, and the reaction is usually carried out under cooling to heating.
Process 3 The object compound (la) or a salt thereof can be prepared by subjecting a compound (Ib) or a salt thereof to elimination reaction of the acyl group.
This reaction is carried out in accordance with a conventional method such as hydrolysis, reduction or the like.
The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid. Suitable base may include an inorganic base and an organic base such as an alkali metal [e.g.
sodium, potassium, etc.], an alkaline earth metal magnesium, calcium, etc.], the hydroxide or carbonate or bicarbonate thereof, trialkylamine trimethylamine, triethylamine, etc.], WO 01/60846 PCT/JP01/01204 26 picoline, 1,5-diazabicyclo[4.3.0]non-5-ene, 1,4diazabicyclo[2.2.2]octane, 1,8-diazabicyclo[5.4.0]undec-7-ene, or the like.
Suitable acid may include an organic acid formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.] and an inorganic acid [e.g.
hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.]. The elimination using Lewis acid such as trihaloacetic acid trichloroacetic acid, trifluoroacetic acid, etc.] or the like is preferably carried out in the presence of cation trapping agents anisole, phenol, etc.].
The -reaction is usually carried out in a solvent such as water, an alcohol methanol, ethanol, etc.], methylene chloride, tetrahydrofuran, a mixture thereof or any other solvent which does not adversely influence the reaction. A liquid base or acid can be also used as the solvent. The reaction temperature is not critical and the reaction is usually carried out under cooling to warming.
The reduction method applicable for the elimination reaction may include chemical reduction and catalytic reduction.
Suitable reducing agents to be used in chemical reduction are a combination of metal tin, zinc, iron, etc.] or metallic compound chromium chloride, chromium acetate, etc.] and an organic or inorganic acid formic acid, acetic acid, propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, etc.].
Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts platinum plate, spongy platinum, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.], palladium catalysts spongy palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, palladium on barium, sulfate, palladium on WO 01/60846 PCT/JP01/01204 27 barium carbonate, etc.], nickel catalysts reduced nickel, nickel oxide, Raneynickel, etc.], cobalt catalysts reduced cobalt, Raney cobalt, etc], iron catalysts reduced iron, Raney iron, etc], copper catalysts reduced copper, Raney copper, Ullman copper, etc.] and the like.
The reduction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, N,N-dimethylformamide, or a mixture thereof. Additionally, in case that the above-mentioned acids to be used in chemical reduction are in liquid, they can also be used as a solvent. Further, a suitable solvent to be used in catalytic reduction may be the above-mentioned solvent, and other conventional solvent such as diethyl ether, dioxane, tetrahydrofuran, etc., or a mixture thereof.
The reaction temperature of this reduction is not critical and the reaction is usually carried out under cooling to warming.
Process 4 The object compound (Id) or a salt thereof can be prepared by reacting the compound (Ic) or its reactive derivative at the amino group or a salt thereof with the compound of the formula: R- OH (V) (wherein R4 is acyl group) or its reactive derivative at the carboxy group or a salt thereof.
Suitable reactive derivative at the carboxy group of the compound may include an acid halide, an acid anhydride, an activated amide, an activated ester, and the like. Suitable examples of the reactive derivatives may be an acid chloride; an acid azide; a mixed acid anhydride with an acid such as substituted phosphoric acid dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, etc.], WO 01/60846 PCT/JP01/01204 28 dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, sulfonic acid methanesulfonic acid, etc.], aliphatic carboxylic acid acetic acid, propionic acid, butyric acid, isobutyric acid, pivaric acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid trichloroacetic acid, etc.]; or aromatic carboxylic acid benzoic acid, etc.]; a symmetrical acid, anhydride; an activated amide with imidazole, 4-substituted imidazole, dimethylpyrazole, triazole, tetrazole or l-hydroxy-iHbenzotriazole; or an activated ester cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [(CH 3 2 N=CH-] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4dinitrophenyl ester, trichlorophenyl ester, pentachloropentyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, etc.], or an ester with a N-hydroxy compound [e.g.
N,N-dimethylhydroxylamine, l-hydroxy-2-(1H)-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, l-hydroxy-lHbenzotriazole, etc.], and the like. These reactive derivatives can optionally be selected from them according to the kind of the compound to be used.
Suitable salts of the compound and its reactive derivative can be referred to the ones as exemplified for the polypeptide compound The reaction is usually carried out in a conventional solvent such as water, alcohol methanol, ethanol, etc.], acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,Ndimethylformamide, pyridine or any other organic solvent which does not adversely influence the reaction. These conventional solvent may also be used in a mixture with water.
In this reaction, when the compound is used in a free acid form or its salt form, the reaction is preferably carried WO 01/60846 PCT/JP01/01204 29 out in the presence of a conventional condensing agent such as N,N'-dicyclohexylcarbodiimide; N-cyclohexyl-N'-morpholinoethylcarbodiimide; N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide; N,N'-diethylcarbodiimide; N,N'-diisopropylcarbodiimide; N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide; N,N-carbonylbis-(2-methylimidazole); pentamethyleneketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine, ethoxyacetylene; l-alkoxy-2-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride); phosphorus trichloride; thionyl chloride; oxalyl chloride; lower alkyl haloformate ethyl chloroformate, isopropyl chloroformate, etc.]; triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt; (m-sulfophenyl)isoxazolium hydroxide intramolecular salt; 1-(p-chlorobenzenesulfonyloxy)-6-chloro-lH-benzotriazole; so-called Vilsmeier reagent prepared by the reaction of N,N-dimethylformamide with thionyl chloride, phosgene, trichloromethyl chloroformate, phosphorous oxychloride, methanesulfonyl chloride, etc.; or the like.
The reaction may also be carried out in the presence of an inorganic or organic base such as an alkali metal carbonate, alkali metal bicarbonate, tri(lower)alkylamine triethylamine, diisopropylethylamine, etc.), pyridine, di(lower)alkylaminopyridine 4-dimethylaminopyridine, etc.), N-(lower)alkylmorpholine, N,N-di(lower)alkylbenzylamine, or the like.
The reaction temperature is not critical, and the reaction is usually carried out under cooling to warming.
The compounds obtained by the above Processes 1 to 4 can be isolated and purified by a conventional method such as pulverization, recrystallization, column-chromatography, WO 01/60846 PCT/JP01/01204 high-performance liquid chromatography (HPLC), reprecipitation, desalting resin column chromatography, or the like.
The compounds obtained by the above Processes 1 to 4 may be obtained as its solvate hydrate, ethanolate, etc.), and its solvate hydrate, ethanolate, etc.) is included within the scope of the present invention.
It is to be noted that each of the polypeptide compound (I) may include one or more stereoisomer such as optical isomer(s) and geometrical isomer(s) due to asymmetric carbon atom(s) and double bond(s) and all such isomers and the mixture thereof are included within the scope of the present invention.
The polypeptide compound or a salt thereof may include solvated compound hydrate, ethanolate, etc.].
The polypeptide compound or a salt thereof may include both its crystal form and non-crystal form.
It should be understood that the polypeptide compound (I) of the present invention may include the prodrug form.
The patent applications and publications cited herein are incorporated by reference.
In order to show the usefulness of the polypeptide compound of the present invention, the biological data of the representative compound is explained in the following.
Biological property of the polypeptide compound of the present invention Test (Antimicrobial activity): SIn vitro antimicrobial activity of the object compound of Examples 4, 9, 25 and 30 disclosed later was determined by MIC Sin mouse serum as described below.
Test Method: The MIC s in mouse serum were determined by the microdilution method using ICR mouse serum buffered with 20 mM HEPES buffer (pH WO 01/60846 PCT/JP01/01204 31 7.3) as a test medium. Inoculum suspension of 106 cells/ml were prepared by a hemocytometric procedure and diluted to obtain an inoculum size of approximately 1.0 x 10 3 cells/ml. Microplates were incubated at 37°C for 24 hours in 5% CO 2 The MIC s were defined as the lowest concentrations at which no visible growth was observed.
Test Result: MIC (Rg/ml) Test organism Candida albicans FP-633 Test compound The object compound of 0.3 Example 4 The object compound of 0.3 Example 9 The object compound of 0.3 Example The object compound of 0.3 Example From the test result, it is realized that the polypeptide compound of the present invention has an antimicrobial activity (especially, antifungal activity).
In more details, the polypeptide compound of the present invention have an antifungal activity, particularly against the following fungi.
Acremonium; Absidia Absidia corymbifera, etc); Aspergillus Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus, Aspergillus versicolor, etc); Blastomyces WO 01160846 PCT/JPOI/01204 32 Blastomyces derrnatiticlis, etc); Candida Candifda albicans, Candida glabrata, Candida guillierrnondii, Candida kefyr, Candida krusei, Candida pa-rapsilosis, Candida stellatoidea, Candida tropi calls, candida utilis, etc.); Cladosporiun Ciadosporiun triahlides, etc); Coacidioldes Coccidioides immitis, etc); Cryptocoacus Cryptococcus neoformans, etc); Cunningharnelia Cunningharnella elegans, etc); Dermnatophyte; Exophiala Exophiala dermatitidis, Exophiala spinifera, etc); Epidermophyton Epidexrmophyton fioccosum, etc); Fonsecaea Fonsecaea pedrosci, eta); Fusariun Fusariun solani, etc); Geotrichurn Geotrichun candiddun, etc); Histoplasina Histopiasma capsulatun var. capsulat ur, etc) Malassezia Malassezia furfur, etc); Microsporun Microsporun canis, Miarosporun gypseun, etc); Mucor; Paracocaidioldes Paracoacidicides brasilieisis, etc); Penialiliun Penicilliun marneffei, etc); Phialophora; Pneurnocystis Pneurnocystis carinii, etc); Pseudallescheria Pseudallesaheria boydii, etc); Rhizopus Rhizopus microsporus var. rhizopodiformis, Rhizopus oryzae, etc); Sacaharornyces Saccharornyces cerevisiae, etc)/ Scopulariopsis; Sporothrix Sporothrix sahenckii, etc); Trichophyton Triahophyton mentagrophytes, Triahophyton ru-brua, etc); Triahosporon Triahosporon asahii, Triahosporon cutaneun, etc).
WO 01/60846 PCT/JP01/01204 33 The above fungi are well-known to cause various infection diseases in skin, eye, hair, nail, oral mucosa, gastrointestinal tract, bronchus, lung, endocardium, brain, meninges, urinary organ, vaginal protion, oral cavity, ophthalmus, systemic, kidney, bronchus, heart, external auditory canal, bone, nasal cavity, paranasal cavity, spleen, liver, hypodermal tissue,.
lymph doct, gastrointestine, articulation, muscle, tendon, interstitial plasma cell in lung, blood, and so on.
Therefore, the polypeptide compound of the present invention are useful for preventing and treating various infectious diseases, such as dermatophytosis trichophytosis, etc), pityriasis versicolor, candidiasis, cryptococcosis, geotrichosis, trichosporosis, aspergillosis, penicilliosis, fusariosis, zygomycosis, sporotrichosis, chromomycosis, coccidioidomycosis, histoplasmosis, blastomycosis, paracoccidioidomycosis, pseudallescheriosis, mycetoma, mycotic keratitis, otomycosis, pneumocystosis, fungemia, and so on.
The combination use of azoles such as fluconazole, voriconazole, itraconazole, ketoconazole, miconazole, ER 30346 and SCH 56592; polyenes such as amphotericin B, nystatin, liposamal and lipid forms thereof such as Abelcet, AmBisome, and Amphocil; purine or pyrimidine nucleotide inhibitors such as flucytosine; or polyxins such as nikkomycines, in particular nikkomycine Z or nikkomycine X; other chitin inhibitors; elongation factor inhibitors such as sordarin and analogs thereof; mannan inhibitors such as predamycin, bactericidal/permeability-inducing (BPI) protein products such as XMP.97 or XMP.127; or complex carbohydrate antifungal agents such as CAN-296; or the combination use of immunosuppressant such as tacrolimus with the polypeptide compound or a salt thereof is effective against above infectious diseases.
WO 01/60846 PCT/JP01/01204 34 The pharmaceutical composition of the present invention can be used in the form of a pharmaceutical preparation, for example, in solid, semisolid or liquid form, whi.ch contains the polypeptide compound or a pharmaceutically acceptable salt thereof, as an active ingredient in admixture with an organic or inorganic carrier or excipient which is suitable for rectal; pulmonary (nasal or buccal inhalation); ocular; external (topical); oral administration; parenteral (including subcutaneous, intravenous and intramuscular) administrations; insufflation (including aerosols from metered dose inhalator); nebulizer; or dry powder inhalator.
The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable carriers in a solid form such as granules, tablets, dragees, pellets, troches, capsules, or suppositories; creams; ointments; aerosols; powders for insufflation; in a liquid form such as solutions, emulsions, or suspensions for injection; ingestion; eye drops; and any other form suitable for use. And, if necessary, there may be included in the above preparation auxiliary substance such as stabilizing, thickening, wetting, emulsifying and coloring agents; perfumes or buffer; or any other commonly may be used as additives.
The polypeptide compound or a pharmaceutically acceptable salt thereof is/are included in the pharmaceutical composition in an amount sufficient to produce the desired antimicrobial. effect upon the process or condition of diseases.
For applying the composition to humans, it is preferable to apply it by intravenous, intramuscular, pulmonary, oral administration, eye drop administration or insufflation. While the dosage of therapeutically effective amount of the polypeptide compound varies from and also depends upon the age and condition of each individual patient to be treated, in the case of intravenous administration, a daily dose of 0.01-400 mg of the polypeptide compound per kg weight of human being in the case of intramuscular administration, a daily dose of 0.1-20 mg of the WO 01/60846 PCT/JP01/01204 polypeptide compound per kg weight of human being, in case of oral administration, a daily dose of 0.5-50 mg of the polypeptide compound per kg weight of human being is generally given for treating or preventing infectious diseases.
Especially in case of the treatment of prevention of Pneumocystis carinii infection, the followings are to be noted.
For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of an aerosol spray presentation form pressurized as powders which may be formulated and the powder compositions may be inhaled with the aid of an insufflation powder inhaler device. The preferred delivery system for inhalation is a metered dose inhalation aerosol, which may be formulated as a suspension or solution of compound in suitable propellants such as fluorocarbons or hydrocarbons.
Because of desirability to directly treat lung and bronchi, aerosol administration is a preferred method of administration.
Insufflation is also a desirable method, especially where infection may have spread to ears and other body cavities.
Alternatively, parenteral administration may be employed using drip intravenous administration.
For administration by intravenous administration, the preferred pharmaceutical composition is the lyophilized form containing the polypeptide compound or its pharmaceutically acceptable salt.
The amount of the polypeptide compound or its pharmaceutically acceptable salt contained in the composition for a single unit dosage of the present invention is 0.1 to 400 mg, more preferably 1 to 200 mg, still more preferably 5 to 100 mg, specifically 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 75, 80, 85, 90, 95 and 100 mg.
The present invention further provides the following ones.
An article of manufacture, comprising packaging material WO 01/60846 PCT/JP01/01204 36 and the compound identified in the above contained within said packaging material, wherein said the compound is therapeutically effective for preventing or treating infectious diseases caused by pathogenic microorganism, and wherein said packaging material comprises a label or a written material which indicates that said compound can or should be used for preventing or treating infectious diseases caused by pathogenic microorganism.
A commercial package comprising the pharmaceutical composition containing the compound identified in the above and a written matter associated therewith, wherein the written matter states that the compound can or should be used for preventing or treating infectious diseases caused by pathogenic microorganism.
The following Preparations and Examples are given for the purpose of illustrating the present invention in more detail.
WO 01/60846 PCT/JP01/01204 37 Preparation 1 To a solution of l-N-t-butyloxycarbonyl-4hydroxypiperidine (5.0 g) in dimethylformamide (DMF) (25 ml) was portionwise added sodium hydride (60% in oil) (1.29 g) with stirring under ice-cooling. The mixture was successively stirred at ambient temperature for 30 minutes, stirred at 60 0
C
for 1 hour and cooled with an ice bath. To the reaction mixture was added 1,5-dibromopentane (6.72 ml), and the mixture was stirred at ambient temperature for 3 hours. The reaction solution was poured into water (100 ml) and extracted twice with a mixture of ethyl acetate (80 ml) and n-hexane (30 ml). The extract was washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (5:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give butoxycarbonylpiperidine (2.44 g).
NMR (CDC1 3 1.46 (9H, 1.50-1.70 (6H, 1.70-1.96 (4H, 3.00-3.15 (2H, 3.35-3.50 (5H, m), 3.70-3.90 (2H, m) APCI MASS 250 (M+-101) Preparation 2 To a solution of butoxycarbonylpiperidine (2.44 g) in methanol (13 ml) was added 28% sodium methoxide methanol solution (14.2 ml), and the mixture was stirred under reflux for 4 hours. The reaction mixture was evaporated in vacuo. The resulting residue was chromatographed on silica gel (250 ml) eluting with a mixture of n-hexane and ethyl acetate (5:1 The fractions containing the object compound were collected and evaporated under reduced pressure to give 4-(5-methoxypentyloxy)-1-N-t-butoxycarbonylpiperidine (1.97 g).
NMR (CDC1 3 1.45 (9H, 1.45-1.95 (10H, 3.03 (1H, WO 01/60846 PCT/JP01/01204 38 dd, J=3.47 and 9.20Hz), 3.10 (1H, dd, J=3.47 and 9.20Hz), 3.44 (3H, 3.34-3.50 (5H, 3.70-3.85 (2H, m) APCI MASS 202 (M+-l01) Preparation 3 To a solution of 4-(5-methoxypentyloxy)-1-N-tbutoxycarbonylpiperidine (1.97 g) in ethyl acetate (20 ml) was added 4N-hydrogen chloride ethyl acetate solution (16.3 ml), and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was evaporated in vacuo. The resulting residue was dissolved in a mixture of dichloromethane and methanol (10:1; ml:5 ml). To this solution was added 1N-sodium hydroxide ml) with stirring. The organic layer was separated and evaporated under reduced pressure to give methoxypentyloxy)-piperidine (0.62 g).
NMR (CDC13, 1.25-1.50 (2H, 1.50-1.75 (6H, m), 1.9-2.10 (2H, 2.70-2.90 (2H, 2.95-3.20 (2H, 3.33 (3H, 3.35-3.50 (5H, m) APCI MASS 202 (M Preparation 4 A solution of 4-fluorobenzonitrile (0.3B methoxypentyloxy)piperidine (0.62 g) and potassium carbonate (0.87 g) in DMF (8 ml) was stirred at 90-95 0 C for 6 hours. The reaction mixture was poured into water (50 ml) and extracted twice with a mixture of ethyl acetate and n-hexane (50ml:20 ml) The extracts were combined, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo.
The resulting residue was chromatographed on silica gel (100 ml) eluting with a mixture of n-hexane and ethyl acetate (5:1 v/v 2:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give 4- (5-methoxypentyloxy)-N-(4-cyanophenyl)piperidine (294 mg).
NMR (CDC1 3 1.35-1.55 (2H, 1.55-1.75 (5H, m), WO 01/60846 PCT/JP01/01204 39 1.85-2.05 (2H, 3.13 (1H, dd, J=3.47 and 9.20Hz), 3.17 (1H, dd, J=3.47 and 9.20Hz), 3.33 (3H, s), 3.35-3.75 (8H, 6.85 (2H, d, J=9.01Hz), 7.47 (2H, d, J=8.96Hz) APCI MASS 303 (M Preparation A solution of 4-(5-methoxypentyloxy)-N-(4cyanophenyl)piperidine (294 mg) and thiosemicarbazide (0.68 g) in toluene (20 ml) and trifluoroacetic acid (10 ml) was stirred at 60-65 C for 7 hours. After cooling, the reaction mixture was poured into a mixture of water (100 ml) and ethyl acetate (200 ml) and adjusted to pH 10 with iN-sodium hydroxide. The mixture was dissolved in a mixture of THF (50 ml) and methanol (10 ml).
The organic layer was separated, washed with saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and evaporated in vacuo. The resulting precipitate was washed with isopropyl ether and dried in vacuo to give 2-amino-5-[4-[4piperidin-l-yl]phenyl]-l,3,4-thiadiazole (1.29 g).
NMR (CDC13+CD 3 0D, 1.30-1.50 (2H, 1.50-1.80 (6H, m), 1.90-2.10 (2H, 2.9-3.10 (2H, 3.34 (3H, s), 3.35-3.70 (7H, 6.93 (2H, d, J=8.91Hz), 7.63 (2H, d, J=8.83Hz) APCI MASS 377 (M Preparation 6 To a suspension of 2-amino-5-[4-[4-(5methoxypentyloxy)piperidin-l-yl]phenyl]-1,3,4-thiadiazole (1.29 g) in ethanol (20 ml) was added ethyl 4-bromoacetylbenzoate (1.39 g) and stirred at reflux for 5 hours. The reaction mixture was cooled and poured into diisopropyl ether (IPE) (60 ml) The resulting precipitate was collected by filtration and dried. To a suspension of the precipitate in xylene (40 ml) was added trifluoroacetic acid (4 ml), and the mixture was stirred at WO 01/60846 PCT/JP01/01204 reflux (130°C) for 5 hours. The reaction mixture was cooled and poured into IPE (300 ml) The resulting precipitate was filtered and dried to give 4-[2-[4-[4-(5-methoxypentyloxy)piperidin- 1-yl]phenyl]imidazo[2,1-b][1,3,4]thiadiazol-6-yl]benzoic acid ethyl ester trifluoroacetic acid salt (2.01 g).
NMR (CDC1 3 1.42 (3H, t, J=7.12Hz), 1.45-1.75 (6H, m), 1.85-2.10 (2H, 2.30-2.50 (2H, 3.36 (3H, s), 3.35-3.55 (5H, 3.60-3.80 (2H, 4.40 (2H, q, J=7.14Hz), 7.57 (2H, d, J=8.78 Hz), 7.84 (2H, d, J=8.40Hz), 7.91 (2H, d, J=8.79Hz), 8.13 (1H, s) APCI MASS 549 Preparation 7 To a solution of 4-[2-[4-[4-(5-methoxypentyloxy)piperidin-l-yl]phenyl]imidazo[2,l-b] [1,3,4]thiadiazol-6yl]benzoic acid ethyl ester trifluoroacetic acid salt (2.01 g) in a mixture of methanol (40 ml) and tetrahydrofuran (20 ml) was added 4N-NaOH (20 ml), and the mixture was refluxed for 6 hours.
The reaction mixture was cooled, poured into water (200 ml) and adjusted to pH 2 with conc. HC1. The resulting precipitate was collected by filtration, washed in turn with water, isopropyl alcohol (30 ml) and IPE (50 ml) to give methoxypentyloxy)piperidin-1-yl]phenyl]imidazo[2,1-b] [1,3,4]thiadiazol-6-yl]benzoic acid (1.28 g).
ESI MASS (Negative): 519.2 Preparation 8 To a solution of 4-[2-[4-[4-(5-methoxypentyloxy)piperidin-1-yl]phenyl]imidazo[2,1-b][1,3,4]thiadiazol-6yl]benzoic acid (1.28 g) and l-hydroxybenzotriazole (465 mg) in dichloromethane (50 ml) was added 1-ethyl-3-(3'dimethylaminopropyl)carbodiimide hydrochloride (WSCD-HC1) (943 mg), and the mixture was stirred overnight at ambient temperature.
The reaction mixture was evaporated in vacuo. To the resulting precipitate was added water (50 ml) and filtered. The precipitate was washed with water and IPE (50 ml) and dried under WO 01160846 PCT/JPOI/01204 41 reduced pressure for 3 hours to give methoxypentyloxy) piperidin-1-yll phenyl] imidazo- [1,3,4]thiadiazol-6--yl]benzoic acid benzotriazol-1-yl ester (1.26 g).
IR 1774.2, 1708,6, 1604.5, 1471.4, 1365.4, 1230.4 cm.J' NMR (CDC1 3 6) 1.30-1.80 (8H1, in), 1.85-2.10 (2H, mn), 3.05-3.30 (2H, mn), 3.33 (3H, 3.35-3.55 (4H, mn), 3.55-3.75 (2H, mn), 6. 94 (2H, cd, J=8.94Hz), 7.30-7.60 (3H1, in), 7.73 (2H, d, J=8.79Hz), 8.00-8.20 mn), 8.30 (2H, d, J=8 .46Hz) ESI MASS (Positive): 660.1 (M++Na) The Starting Compounds used and the Object Compounds obtained in the following Preparation 9 is given in the table as below, in which the formula of the starting compound is in the upper column and the formula of the object compound are in the lower column, respectively.
WO 01/60846 PCT/JP01/01204 Preparation No. Formula
OH
HO O H3C.., N A NHBoc H2N O 0 HN OH HO H 0 CH3 ?D
'OH
NaO3SO b
HC
9
OH
HO 0 H3C,,N. 1 N NHBoc No- o HDOOH HO H 0 CH3
SOHOH
Na03SO k
HO
Preparation 9 To a solution of a mixture of the starting compound (5.4 2-oxo-1,3-diacetoxypropane (4.85 g) and acetic acid (0.78 ml) in a mixture of methanol (80 ml) and dimethylformamide ml) was added sodium cyanoborohydride (1.71 g) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight. The reaction mixture was concentrated in vacuo. To the resulting residue was added pH 6.86 standard buffer solution (100 ml) and acetonitrile (20 ml), and the solution was adjusted to pH 8.5 with IN sodium hydroxide. The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (400 ml) eluting in turn with water, 20% acetonitrile in water and 25% acetonitrile in water. The fractions containing WO 01/60846 PCT/JP01/01204 43 the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (4.44 g).
IR (KBr): 1632, 1516, 1452, 1273, 1248 cm-1 NMR (DMSO-d 6 0.98 (3H, d, J=6.88Hz), 1.11 (3H, d, J=5.64Hz), 1.36 (9H, 1.40-2.00 (6H, 2.50-2.95 (4H, 3.30-3.55 (2H, 3.65-4.45 (16H, m), 4.70-4.85 (2H, 5.36 (1H, 6.71 (1H, d, J=8.05Hz), 6.77 (1H, d, J=8.29Hz), 6.99 (1H, 7.30-7.45 m) APCI MASS (m/z)(Positive): 1175.4 (M++Na) Elemental Analysis Calcd. for C 50
H
72
N
8 0 21 S-5H 2 0: C 46.80, H 6.52, N 8.73 Found: C 47.06, H 6.44, N 8.54 Preparation To a solution of trans-4-methylcyclohexanol (4.55 g) in ethyl acetate (50 ml) were added successively triethylamine (7.22 ml) and methanesulfonyl chloride (3.38 ml) with stirring under ice-water bath. The mixture was stirred at the same temperature for 1 hour. To the reaction mixture were added ethyl acetate (50 ml), water (50 ml) and 1N hydrochloric acid (20 ml) with stirring. The organic layer was separated, washed successively with water, saturated aqueous sodium hydrogen carbonate, water and saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo to give trans-4-methylcyclohexyl methanesulfonate (8.36 g).
NMR (CDC1 3 0.90 (3H, d, J=6.46Hz), 0.95-1.70 (5H, m), 1.70-1.85 (2H, 2.00-2.20 (2H, 3.00 (3H, s), 4.50-4.70 (1H, m) Preparation 11 Piperazine (54.9 g) and methanol (5 ml) was stirred at 120 0
C
to melt for 15 minutes. To the solution was dropwise added trans-4-methylcyclohexyl methanesulfonate (33.0 g) and the WO 01/60846 PCT/JP01/01204 44 mixture was stirred at the same temperature for 2 hours. After cooling, to the reaction mixture was added water (150 ml) and extracted three times with a mixture of ethyl acetate (100 ml) and THF (100 ml). The extracts were collected, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (600 ml) eluting with a mixture of dichloromethane, methanol and conc. ammonium hydroxide (4:1:0.1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give cis-l-(4-methylcyclohexyl)piperazine (17.76 g).
NMR (CDC1 3 0.92 (3H, d, J=6.96Hz), 1.40-1.65 (BH, m), 1.65-1.85 (1H, 2.05-2.25 (1H, 2.45-2.60 (4H, 2.85-2.95 (4H, m) APCI MASS (m/z)(Positive): 183.2 Preparation 12 A solution of ethyl 4-[4-(4-methylenecyclohexyl)-lpiperazinyl]benzoate (100 mg) and Iridium black (30 mg) in a mixture of t-butanol (1 ml) and methanol (2 ml) was stirred under atmospheric pressure of hydrogen for 4 hours. The catalyst was filtrated off and the filtrates were evaporated in vacuo. Ethyl 4-[cis-4-(4-methylcyclohexyl)-l-piperazinyl]benzoate and ethyl 4-[trans-4-(4-methylcyclohexyl)-1-piperazinyl]benzoate were obtained in the ratio 5:1-6:1 by thin-layer chromatography.
Preparation 13 To a suspension of l-[4-[5-(4-iodophenyl)-1,3,4thiadiazol-2-yl]phenyl]-4-(4-methylcyclohexyl)piperazine (2 g) in DMF (40 ml) was successively added ethyl formate (0.56 ml), dichlorobis(triphenylphosphine)palladium(II) (0.52 g) and sodium ethoxide ethanol solution (4.43 ml) with stirring and the mixture was stirred at 40 0 C for 2 hours. The reaction mixture was added diisopropyl ether (600 ml) The resulting precipitate was collected by filtration. The precipitate was dissolved THF (200 ml), insoluble materials were filtered off and solution was WO 01/60846 PCT/JP01/01204 concentrated in vacuo. The resulting residue was washed with acetonitrile, and dried to give ethyl methylcyclohexyl)-l-piperazinyl]phenyl]-1,3,4-thiadiazol-2yl]benzoate (1.07 g).
NMR (CDC13, 0.93-0.96 (3H, 1.39-3.37 (18H, 4.42 (2H, q, J=7.1Hz), 6.96 (2H, d, J=8.9Hz), 7.87-8.17 (6H, m) MASS 491.4 Preparation 14 To a solution of methyl triphenylphosphonium bromide (13.7 g) in DMSO (140 ml) was added potassium tert-butoxide (4.31 g) under ice-cooling and the mixture was stirred for 1.5 hours at ambient temperature. After cooling, 1,4dioxaspiro[4,5]decan-8-one (5.0 g) was dropwise added to the solution under ice-cooling and then stirred for 1 hour at room temperature. The reaction mixture was poured into water (300 ml) and extracted twice with ethyl acetate (150 ml). The extracts were washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo to give a residue. The residue was washed with a mixture of hexane and ethyl acetate (5:1 v/v) (300 ml). The resulting precipitates were collected and were chromatographed on silica gel (500 ml) eluting with a mixture of n-hexane and ethyl acetate (5:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give 8-methylene 1,4- (5.56 g).
NMR (CDC1 3 1.70 (4H, t, J=6.46Hz), 2.29 (4H, t, J=6.84Hz), 3.97 (4H, 4.67 (2H, s) Preparation A solution of 8-methylene 1,4-dioxaspiro[4,5]decane (5.55 g) in a mixture of acetone (60 ml) and water (4 ml) and ptoluenesulfonic acid monohydrate (1.37g) was stirred at ambient temperature overnight. To a solution was added p- WO 01/60846 PCT/JP01/01204 46 toluenesulfonic acid monohydrate (1.37 g) and the mixture was stirred at ambient temperature for 8 hours. Ethyl acetate (150 ml) was added to the reaction mixture and the solution was washed in turn with water, saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (150 ml) eluting with a mixture of n-hexane and-ethyl acetate (9:1 The fraction containing the desired compound were collected and evaporated under reduced pressure to give 4-methylenecyclohexan-l-one (0.98 This compound was immediately used as the starting compound for the next step.
Preparation 16 To a solution of 1,4-dioxaspiro[4,5]decan-8-one (96.9 g) in methanol (1 1) was added portionwise sodium borohydride (46.9 g) under ice-cooling. After stirring for 3.5 hours under ice cooling, the reaction mixture was successively stirred for 3 hours at room temperature. Then the solvent was evaporated in vacuo and the residue was purified by column chromatography on silica gel eluting with a mixture of hexane and ethyl acetate The eluted fractions containing the desired product were collected and evaporated in vacuo to give 1,4-dioxaspiro- [4,5]decan-8-ol (97.9 g).
NMR (CDC13, 1.4-2.0 (9H, 3.7-3.9 (1H, 3.95 (4H, s) Preparation 17 To a solution of tert-butyl 4-(5-bromophenyloxy)-1piperidinecarboxylate (12.98 g) in methanol (70 ml) was added 28% sodiummethoxide methanol solution (37.8 ml) and the mixture was stirred under refluxing for 4 hours. After cooling, the reaction mixture was evaporated in vacuo. The resulting residue was chromatographed on silica gel (400 ml) eluting with a mixture of n-hexane and ethyl acetate (5:1 The fractions WO 01/60846 PCT/JP01/01204 47 containing the object compound were collected and evaporated under reduced pressure to give tert-butyl methoxypentyloxy)-1-piperidinecarboxylate (16.31 This compound was immediately used as the starting compound for the next step.
Preparation 18 A solution of ethyl 4-fluorobenzoate (2.30 4-4- (methoxybutyloxymethyl)piperidine trifluroacetate (3.6 g) and potassium carbonate (4.73 g) in DMSO (40 ml) was stirred at 140-150 0 C for 4 hours. The reaction mixture was poured into water (150 ml) and extracted twice with ethyl acetate (80 ml). The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo.
The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (2:1 v/v).
The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl methoxybutyloxymethyl)-l-piperidyl]benzoate (2.76 g).
NMR (CDC13, 1.20-1.45 (5H, 1.50-1.70 (3H, m), 1.70-1.90 (3H, 2.84 (2H, dt, J=2.49 and J=12.8Hz), 3.28 (2H, d, J=6.01Hz), 3.33 (3H, 3.35-3.50 (4H, 3.75-3.90 (2H, 4.32 (2H, q, J=7.11Hz), 6.85 (2H, d, J-9.06Hz), 7.92 (2H, d, J=9.01 Hz) APCI MASS (Positive): 350.4 Preparation 19 To a solution of tert-butyl (methoxypentyloxymethyl)-1-piperidinecarboxylate (1.75 g) in dichloromethane (50 ml) and anisole (4.22 ml) was added trifluoroacetic acid (8.55 ml) under ice-cooling and the mixture was stirred at ambient temperature for 1 hour. The reaction mixture was evaporated in vacuo and azeotropically distilled three times with toluene (30 ml) and dried in vacuo to give 4-5-(methoxypentyloxymethyl)piperidine trifluoroacetate (7.30 WO 01/60846 PCT/JP01/01204 48 g, crude oil). A solution of this compound (1.89 ethyl 4-fluorobenzoate (1.21 and potassium carbonate (2.30 g) in DMSO (20 ml) was stirred at 150°C for 4 hours. The reaction mixture was poured into water (100 ml) and extracted twice with ethyl acetate (80 ml). The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (2:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl methoxypentyloxymethyl)-l-piperidyl]benzoate (1.21 g).
NMR (CDC13, 1.20-1.50 (6H, 1.50-1.70 (5H, m), 1.75-1.90 (3H, 2.84 (2H, dt, J=2.49 and 12.7Hz), 3.28 (2H, d, J=6.02Hz), 3.33 (3H, 3.39 (4H, q, J=6.57Hz), 3.75-3.95 (2H, 4.32 (2H, q, J=7.10Hz), 6.86 (2H, d, J=9.06Hz), 7.90 (2H, d, J-9.00Hz) Preparation A solution of methyl 6-chloronicotinate (25.4 g) and piperazine (38.3 g) in dimethylsulfoxide (125 ml) was heated at 100°C for 2 hours then cooled and diluted with water, followed by extraction with ethyl acetate The combined organic layers were washed with water then dried over magnesium sulfate, filtered and evaporated to give a crude product that was triturated with isopropyl ether-hexane to yield methyl 6-(1piperazinyl)nicotinate (25 g) as a light yellow powder.
NMR (CDC13, 1.81 (1H, 2.94-3.01 (4H, 3.63-3.68 (4H, 3.87 (3H, 6.58 (1H, d, J=9Hz), 8.01 (1H, dd, J=2.4 and 9Hz), 8.79 (1H, d, J=2.4Hz) ESI MASS 222 Preparation 21 A solution of tert-butyl-4-[4-(methoxycarbonyl)phenyl]- 1-piperidinecarboxylate (3.95 g) in a mixture of methanol WO 01/60846 PCT/JP01/01204 49 ml), THF (40 ml) and 4N sodium hydroxide (30 ml) was stirred at 0 C for 2 hours. The reaction mixture was concentrated in vacuo.
To a residue was added water (100 ml) and adjusted to pH 3 with 1N hydrochloric acid. The solution was extracted twice with a mixture of ethyl acetate (100 ml) and THF (50 ml). The extracts was dried over magnesium sulfate and evaporated in vacuo to give 4-[1-(tert-butoxycarbonyl)-4-piperidyl]benzoic acid (3.64g).
NMR (CDC1 3 1.49 (9H, 1.50-1.90 (4H, 2.60-2.95 (3H, 4.10-4.30 (2H, 7.30 (2H, d, J=8.33Hz), 8.05 (2H, d, J=8.27Hz) ESI MASS (m/z)(Negative): 304.1 (M -1) Preparation 22 To a solution of 4-[l-(tert-butoxycarbonyl)-4piperidyl]benzoic acid (3.64 g) and 1-hydroxybenzotriazole (2.73 g) in dichloromethane (40 ml) was added l-ethyl-3-(3'dimethylaminopropyl)carbodiimide hydrochloride (WSCD HC1) (4.56 g) and the mixture was stirred at ambient temperature for 3 hours. To the reaction mixture was added water (20 ml) and an organic layer was separated and washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. A solution of the resulting residue in THF (30 ml) was added to a solution of conc. ammonium hydroxide (10 ml) in THF ml) at ambient temperature and the mixture was stirred at the same temperature for 1 hour. To a reaction mixture was added ethyl acetate (200 ml) and the organic layer was separated, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The precipitate was chromatographed on silica gel (200 ml) eluting with a mixture of dichloromethane and methanol (9:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give tert-hutyl 4-[4- (aminocarbonyl)phenyl]-l-piperidinecarboxylate (3.34 g).
NMR (CDC1 3 1.48 (9H, 1.50-1.90 (4H, 2.60-2.95 (3H, 4.15-4.30 (2H, 5.75-6.25 (1H, broad m), WO 01/60846 PCT/JP01/01204 7.33 (2H, d, J=9.14Hz), 7.76 (2H, d, J=8.28Hz) ESI MASS (m/z)(Positive): 327.3 (M++Na) Preparation 23 To a solution of tert-butyl 4-[4- (aminocarbonyl)phenyl]-l-piperidinecarboxylate (3.34 g) in DMF (18 ml) was dropwise added phosphoryloxychloride (3.07 ml) keeping under 100C with stirring and the mixture was stirred at ambient temperature for 10 minutes. The reaction mixture was poured into a mixture of saturated aqueous sodium carbonate ml) and ice-water (250 ml) with stirring and extracted twice with a mixture of ethyl acetate (200 ml) and hexane (80 ml). The extract was washed twice with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (300 ml) eluting with a mixture of n-hexane and ethyl acetate (4:1 v/v).
The fractions containing the desired compound were collected and evaporated under reduced pressure to give tert-butyl 4-(4cyanophenyl)-1-piperidine carboxylate (3.13 g).
IR (KBr): 2227.4, 1699.0, 1677.8, 1608.3, 1504.2, 1423.2, 1369.2 cm-1 NMR (CDC13, 1.48 1.50-1.90 (4H, 2.60-2.95 (3H, 4.15-4.30 (2H, 7.30 (2H, d, J=8.26Hz), 7.60 (2H, d, J=8.33Hz) APCI MASS (m/z)(Positive): 309.3 (M++Na) Preparation 24 Amixture of cis-1-(4-methylcyclohexyl)piperazine (2.15 g), 4-fluorobenzonitrile (1.72 g) and potassium carbonate (4.89 g) in DMSO (25 ml) was stirred at 140 0 C for 4 hours. The reaction mixture was poured into water (150 ml) and extracted twice with ethyl acetate (150 ml) The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture WO 01/60846 PCT/JP01/01204 51 of n-hexane and ethyl acetate (2:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl 4-[cis-4-(4methylcyclohexyl)-1-piperazinyl]benzonitrile (2.81 g).
NMR (CDC1 3 0.93 (3H, d, J=6.87Hz), 1.40-1.80 (9H, m), 2.15-2.30 (1H, 2.60-2.70 (4H, 3.25-3.35 (4H, 6.86 (2H, d, J=9.06Hz), 7.48 (2H, d, J-9.04Hz) APCI MASS (m/z)(Positive): 284.3 Preparation To a solution of 4-hydroxycyclohexylcyclohexane (25 g) in acetone (250 ml) was added dropwise with stirring 2.67N Jone's regend (77 ml) at 0°C. The mixture was then stirred for 1 hour at 0°C. The organic layer was collected and evaporated. The reaction mixture was added to a mixture of water and diethyl ether.
The organic layer was washed with water, sodium hydrogen carbonate solution and brine. The organic layer was taken and dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to give 4-cyclohexylcyclohexanone (19.57 g).
NMR (CDC1 3 0.8-1.4 (6H, 1.4-1.9 (8H, 1.9-2.15 (2H, 2.15-2.5 (4H, m) MASS 181 Preparation 26 A solution of oxalyl chloride (2.14 ml) in dichloromethane ml) was cooled to -78°C in nitrogen atmosphere, and a solution of dimethylsulfoxide (6 ml) in dichloromethane (6 ml) was added slowly and stirred for 10 minutes at -78 0 C. To the reaction mixture was added a solution of 4'-methoxy-l,l'bi(cyclohexyl)-4-ol (2.6 g) in dichloromethane (26 ml) slowly to maintain the reaction temperature and stirred for 2.5 hours at -40°C. To the reaction mixture was added triethylamine (12.4 ml) slowly. Then the reaction mixture allowed to warm to room temperature. To the reaction mixture was added ammonium chloride WO 01/60846 PCT/JP01/01204 52 solution and ethyl acetate. The organic layer was taken and dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to give 4'-methoxy-l,l'-bi(cyclohexyl)-4-one (1.62 g).
NMR (CDC13, 0.9-2.5 (18H, 3.0-3.2 (1H, 3.35 (3H, s) MASS 233 (M +1) Preparation 27 A solution of 4-(4-hydroxyphenyl)cyclohexanone (5 g) and iodomethane (0.828 ml) in N,N-dimethylformamide (50 ml) was treated with potassium carbonate (4.36 g) at room temperature for 28 hours. Water was poured into the reaction mixture. And the resulting precipitate was collected by filtration and washed with isopropanol and diisopropyl ether to give 4-(4methoxyphenyl)cyclohexanone (6.815 g).
NMR (CDC1 3 1.75-2.35 (4H, 2.4-2.6 (4H, 2.9- 3.1 (1H, 3.80 (3H, 6.87 (2H, d, J=8.7Hz), 7.17 (2H, d, J=8.7Hz) MASS 227 (M++23) Preparation 28 To a solution of 4,4-dimethyl-2-cyclohexen-l-one (10 g) in ethanol (100 ml) was added 10% palladium on carbon (1 and hydrogen gas at atmosphere pressure for 4 hours. To the reaction mixture was filtered. The filtrate was concentrated by evaporation under reduced pressure to give 4,4dimethylcyclohexanone (10.03 g).
NMR (CDC1 3 0.92 (3H, 1.10 (3H, 1.25-1.4 (2H, 1.5-1.75 (4H, 2.3-2.45 (2H, m) MASS 149 (M++23) Preparation 29 Amixture of cis-l-(4-methylcyclohexyl)piperazine (29.6 g), ethyl 4-fluorobenzoate (41.0 g) and potassium carbonate (67.3 WO 01160846 PCT/JPOI/01204 53 g) in DMSO (300 ml) was stirred at 140 0 C for 9 hours. The reaction mixture was poured into water (1.2 1) and extracted twice with ethyl acetate (400 ml) The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (1 1) eluting with a mixture of n-hexane and ethyl acetate (2:1 v/v) The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl 4-[cis-4-(4-methylcyclohexyl)-lpiperazinyllbenzoate (37.64 g).
NMR (CDCl 3 0.93 (3H, d, J=6.87Hz), 1.36 (3H, t, J=7.llHz), 1-40-1.80 (9H, in), 2.15-2.25 (1H, in), 2.60-2.70 (4H, mn), 3.25-3.50 (4H, mn), 4.32 (2H, q, J=7.llHz), 6.86 (2H, d, Jzr8.94Hz), 7.92 (2H, d, J=8.87Hz) ESI MASS (in/z) (Positive) 683.4 (2M++Na) 331.3 The following compounds [Preparations 30 and 31] were obtained according to a similar manner to that of Preparation 29.
Preparation Ethyl 4- (5-methoxypentyloxy) -l-piperidylilbenzoate NMR (CDCl 3 8) 36 (3H, t, J=-7.l2Hz) 1.37-1.75 (8H, in), 1.85-2.05 (2H, in), 3.00-3.18 (2H, in), 3.33, (3H, s), 3.34-3.60 (5H, mn), 3.60-3.75 (2H, mn), 4.32 (2H, q, J=7.llHz), 6.86 (2H, d, J=9.OOHz), 7.90 (2H, d, J=8.93Hz) ESI MASS (in/z) (Positive): 372.3 (M++Na) Preparation 31 Ethyl 4- (6-methoxyhexyloxy) -1-piperidyllhenzoate NMR (CDCl 3 1. 3-1. 8 (3H, m) 1.08-2. 1 (12H, in), 3. 0-3. 2 (2H, in), 3.33 (3H, 3.3-3.5 (5H, mn), 3.6-3.8 (2H, mn), 4.32 (2H, q, J=7.lHz), 6.86 (2H, d, J=9.lHz), WO 01/60846 PCT/JP01/01204 54 7.8-8.0 (2H, m) ESI MASS (m/z)(Positive): 364.33 (M++Na) Preparation 32 To a suspension of 4-[4-(4-methylcyclohexyl)-1piperazinyl]benzohydrazide (12.48 g) and pyridine (11.7 ml) in THF (374 ml) was added 4-iodobenzoyl chloride (11 g) under ice-cooling and the mixture was stirred at the same temperature for 1 hour. The reaction mixture was added water (3700 ml) The resulting precipitate was collected by filtration and dried to give N'-(4-iodobenzoyl)-4-[4-(4-methylcyclohexyl)-1piperazinyl]benzohydrazide (22.45 g).
NMR (DMSO-d 6 0.88-0.92 (3H, 1.42-3.37 (18H, m), 6.99 (2H, d, J=8.9Hz), 7.69 (2H, d, J=8.4Hz), 7.80 (2H, d, J=8.80Hz), 7.92 (2H, d, J=8.44Hz), 10.21 (1H, s), 10.45 (1H, s) MASS 547 The following compounds [Preparations 33 to 46] were obtained according to a similar manner to that of Preparation 32.
Preparation 33 Methyl 4-[2-[4-(4-methylenecyclohexyl)-lpiperazinylbenzoyl]hydrazinocarbonyl]benzoate IR (KBr) 3458, 3253, 2943, 2837, 1722, 1678, 1645, 1608, 1510 cm 1 NMR (DSMO-d 6 1.10-1.50 (2H, 1.60-2.10 (4H, m), 2.15-2.40 (2H, 2.50-2.70 (4H, 3.10-3.40 (4H, 3.80-4.00 (2H, 4.61 (1H, br 6.98 (2H, d, J=8.9Hz), 7.81 (2H, d, J=8.8Hz), 7.95 (4H, q, J=8.6Hz), 10.26 (1H, 10.57 (1H, s) MASS 479 (M 478 477 Preparation 34 WO 01/60846 PCT/JPO/01204 Methyl 4-[2-[4-[4-(4-phenylcyclohexyl)-lpiperazinyllbenzoyl]hydrazinocarbonyl]benzoate APCI MASS 541.4 (M+1) Preparation Methyl 4-[5-[4-(4-phenylcyclohexyl)-lpiperazinyl]phenyl]-1,3,4-thiadiazol- 2 -ylbezoate APCI MASS 539.4 (MS+1) Preparation 36 Methyl 4-[2-[4-[4-(cyclohexylethyl)-lpiperazinyl]benzoyl hydrazinocarbonyl]benzoate NMR (DMSO-d 6 6) 0.7-1.0 (2H, 1.1-1.8 (9H, mn), 2.12 (2H, d, J=7.lHz), 2.4-2.5 (4H, 3.2-3.4 (4H, 3.90 (3H, 6.98 (2H, d, J=9Hz), 7.81 (2H, d, J=9Hz), 8.02 (2H, d, J=8.7Hz), 8.09 (2H, d, J=8.7Hz), 10.26 (1H, 10.58 (1H, s) APCT MASS (m/z)(Positive): 479.4 (MS+l) Preparation 37 4-[2-[4-[4-[4-(7-Methoxyheptyloxy)cyclohexyl]-1piperazinyl benzoyllhydrazinocarbonyl]benzoate IR (Neat): 2933, 2858, 1724, 1682, 1645, 1608, 1279, 1242, 1113 cirC 1 NMR (DSMO-d 6 1.0-2.1 (18H, 2.2-2.4 (1H, 2.52 (4H, 3.0-3.5 (9H, 3.20 (3H, 3.90 (3H, s), 6.9-7.1 (2H, 7.7-7.9 (2H, 8.0-8.2 (4H, m), 10.26 (1W, 10.58 (1H, s) ESI MASS (Positive): 609.5 Preparation 38 4-[2-[4-[4-[4-(8-Methoxyoctyloxy)cyclohexyl]-1piperazinyl]benzoyl hycrazinocarbonyl benzoate IR (Neat) 2931, 2856, 1724, 1680, 1647, 1608, 1520, 1281, 1240, 1113 cm- 1 WO 01160846 PCT/JPO/01204 56 NMR (DSMO-d 6 1.0-2.1 (20H, 2.2-2.4 (1H, 2.5-2.7 (4H, 3.0-3.6 (9H, 3.21 (3H, 3.90 (3H, s), 6.9-7.1 (2H, 7.7-7.9 (2H, 8.0-8.2 (4H, i), 10.26 (1H, 10.58 (1H, s) ESI MASS (n/z)(Positive): 623.5 Preparation 39 Methyl 4-[2-[4-[4-(5-iethoxypentyloxy)-lpiperidyl]benzoyllhydrazinocarbonyl]benzoate NMR (DSMO-d 6 1.20-1.60 (8H, 1.80-1.95 (2H, m), 2.95-3.20 (2H, 3.21 (3H, 3.25-3.80 (7H, m), 3.90 (3H, 6.99 (2H, d, J=8.92Hz), 7.80 (2H, d, J=8.76Hz), 8.03 (2H, d, J=8.56Hz), 8.09 (2H, d, J=0.54Hz), 10.24 (1H, 10.57 (1H, s) ESI MASS (n/z)(Positive): 520.3 (M++Na) Preparation Methyl 4-[2-[4-[4-(6-methoxyhexyloxy)-lpiperidyl]benzoyl]hydrazinocarbonyllbenzoate NMR (CDC1 3 1.3-2.1 (12H, 3.0-3.2 (2H, 3.33 (3H, 3.3-3.5 (5H, 3.5-3.7 (2H, in), 3.94 (31, s), 6.83 (2H, d, J=9.OHz), 7.74 (2H, d, J=8.9Hz), 7.90 (2H, d, J=8.5Hz), 8.04 (2H, d, J=8.5Jz), 9.52 (1H, d, 10.11 (1H, d, J=5.2Hz) APCI MASS (Positive): 512.40 Preparation 41 Methyl 4- (4-methoxybutoxyethyl) -1piperidyl]benzoyl]hydrazinocarbonyl benzoate NMR (CDCl 3 1.2-1.9 (9H, 2.7-2.9 (2H, 3.2-3.5 (6H, 3.33 3.8-4.0 (2H, 3.94 (3H, s), 6.84 (2H, d, J=9.OHz), 7.74 (21, d, J=8.9Hz), 791 (2H, d, J=8.4Hz), 8.06 (2H, d, J=8.5Hz), 9.40 (1H, d, 9.96 (1H, d, J 5.6Hz) APOT MASS (Positive): 497.93 WO 01160846 PCT/JPO/01204 57 Preparation 42 Methyl 4-[2-[4-[4-(5-methoxypentyoxymethyl)-lpiperidyl]benzoyllhydrazinocarbonyl]benzoate NMR (CDCl 3 1.2-1.9 (11H, 2.7-2.9 (2H, 3.2-3.5 (6H, 3.33 (3H, 3.8-4.0 (2H, 3.94 (3H, s), 6.83 (2H, d, J=9.0Hz), 7.74 (21, d, J=8.8Hz), 7.90 (2H, d, J=8.4Hz), 8.05 (2H, d, J=8.4Hz), 9.47 (lH, d, J=4.9Hz), 10.06 (1H, d, J=5.3Hz) ESI MASS (Positive): 534.4 (M++Na) Preparation 43 Methyl 4-[2-[6-[4-(4-methylcyclohexyl)-1-piperazinyl-3pyridyl]carbonyl]hydrazinocarbonyl]benzoate NMR (CDCl 3 0.94 (3H, d, J=6.9Hz), 1.4-2.0 (9H, m), 2.1-2.3 (1H, 2.6-2.7 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.62 (1H, d, J-9.1Hz), 7.8-8.0 (3H, 8.13 (2H, d, J=8.4Hz), 8.69 (1H, d, J=2.3Hz) APCI MASS (i/z)(Positive): 480.27 (M+1) Preparation 44 Methyl 4-[2-[6-[4-(4-methylcyclohexyl)-1-piperazinyl-3pyridyl]carbonyl]hydrazinocarbonyl]benzoate NMR (CDCl 3 0.8-2.0 (12H, 2.2-2.4 (1H, 2.6-2.8 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.62 (1H, d, J=9.2Hz), 7.8-8.0 (3H, 8.14 (2H, d, J=8.5Hz), 8.69 (1H, d, J=2.3Hz) APCT MASS (i/z)(Positive): 480.20 (M+l) PrepQaration Methyl 4-[2-[6-[4-(4-ethylcyclohexyl)-1-piperazinyl-3pyridyl]carbonyl]hydrazinocarbonyl]benzoate NMR (CDCl 3 0.8-1.0 (5H, 1.2-2.0 (9H, 2.1-2.3 (1H, 2.5-2.7 (4K, 3.6-3.8 (4H, 3.96 (3H, 6.62 (1H, d, J=9-lHz), 7.8-8.0 (3H, 8.13 (2H, WO 01/60846 PCT/JP01/01204 58 d, J=8.3Hz), 8.6-8.7 (1H, m) APCI MASS (m/z)(Positive): 494.20 (M +1) Preparation 46 Methyl 4-[2-[6-[4-(4-ethylcyclohexyl)-l-piperazinyl-3pyridyl]carbonyl]hydrazinocarbonyl]benzoate NMR (CDC1 3 0.8-2.0 (14H, 2.2-2.4 (1H, 2.6-2.8 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.63 (1H, d, J=9.2Hz), 7.8-8.0 (3H, 8.0-8.2 (2H, 8.6-8.7 (1H, m) APCI MASS (m/z)(Positive): 494.20 Preparation 47 A suspension of N'-(4-iodobenzoyl)-4-[4-(4methylcyclohexyl)-1-piperazinyl]benzohydrazide (22.95 g) in pyridine (459 ml) was treated with phosphorus pentasulfide (11.2 g) and stirred at 120 0 C for 2.5 hours. The reaction mixture was added a solution of sodium hydroxide (510 g) in water (9200 ml) The resulting precipitate was collected, washed with acetone.
The powder was recrystallized from THF (800 ml) and dried to give 1-[4-[5-(4-iodophenyl)-1,3,4-thiadiazol-2-yl]phenyl]-4-(4methylcyclohexyl)piperazine (16.42 g).
NMR (CDC13, 0.93-0.96 (3H, 1.47-3.36 (18H, 6.95 (2H, d, J=9.0Hz), 7.68-7.90 (6H, m) MASS 545 (M +l) The following compounds [Preparations 48 to 55] were obtained according to a similar manner to that of Preparation 47.
Preparation 48 Methyl 4-[5-[4-[4-(cyclohexylmethyl)-lpiperazinyl]phenyl]-1,3,4-thiadiazol-2-yl]benzoate NMR (CDC1 3 0.8-1.05 (2H, 1.1-2.0 (9H, 2.22 (2H, d, J=7Hz), 2.58 (4H, br 3.33-3.38 (4H, 3.96 WO 01160846 PCT/JPO/01204 59 (3H, 6.96 (2H, d, J=9Hz), 7.89 (2H, d, J=9Hz), 8.06 (2H, d, J 8.6Hz), 8.15 (2H, d, J=8.6Hz) APCI MASS (i/z)(Positive): 477.47 Preparation 49 Methyl 4-15- [4-4-(6-methoxyhexyloxy)-lpiperidyllphenyl]-l,3,4-thiadiazol-2-yl]befzoate NMR (CDCl 3 1.3-2.1 (12H, 3.0-3.2 (2H, 3.33 (3H, 3.3-3.6 (5H, 3.6-3.8 (2H, 3.96 (3H, s), 6.96 (2H, d, J=8.9Hz), 7.88 (2H, d, J=8.6Hz), 8.06 (2H, d, J=8.6Hz), 8.15 (2H, d, J8.6Hz) APCI MASS (m/z)(Positive): 510.47 Preparation Methyl 4-[5-[4-[4-(4-methoxybutoxyiethyl)-lpiperidyl]phenyl -1,3,4-thiadiazol-2-yl]benzoate NMR (CDCl 3 1.2-2.0 (9H, 2.8-3.0 (2H, 3.3-3.5 (6H, 3.34 (3H, 3.8-4.0 (2H, 3.96 (3H, s), 6.95 (2H, d, J=9.0Hz), 7.88 (2H, d, J=8.9Hz), 8.2 (4H, .m) APCI MASS (i/z)(Positive): 496.27 (M+Il) Preparation 51 Methyl 4-[5-[4-[4-(5-methoxypentyloxymethyl)-lpiperidyllphenyl]-1,3,4-thiadiazol-2-yl]belZoate NMR (CDCl 3 1.2-2.0 (11H, 2.7-2.9 (2H, 3.2-3.5 (6H, 3.34 (3H, 3.8-4.0 (2H, 3.96 (3H, s), 6.95 (2H, d, J 9.0Kz), 7.88 (2H, d, J=8.9Hz), 8.2 (4H, m) APCI MASS (Positive) 510.40 Preparation 52 Methyl 4-[5-[6-[4-(4-methylcyclohexyl)-l-piperazifylV 3-pyridyll-1,3,4-thiadiazol-2-y1]benzoate NMR (CDC1 3 0.95 (3H, d, J=6.9Hz), 1.4-1.8 (9H, i), WO 01160846 PCT/JPO/01204 2.1-2.3 (1H, 2.6-2.8 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.72 (1H, d, J=9.lHz), 8.0-8.2 (5H, 8.71 (11, d, J=2.4Hz) APCI MASS (m/z)(Positive): 478.13 (M+1) Preparation 53 Methyl 4-[5-[6-[4-(4-ethylcyclohexy1)-l-piperazifl1- 3-pyridyl]-1,3,4-thiadiazol-2-yl]benzoate NMR (CDC1 3 0.8-2.0 (12H, 2.2-2.4 (11, 2.6-2.3 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.72 (11, d, J=9.OHz), 8.0-8.2 (5H, 8.71 (1H, d, J=2.3Hz) ESI MASS (m/z)(Positive): 478.3 Preparation 54 Methyl 4-[5-[6-[4-(4-ethylcyclohexyl)-l-piperazinyl]-3pyridyl]-1,3,4-thiadiazol-2-yllbenzoate NMR (CDC1 3 0.8-1.0 (5H, 1.2-1.9 (9H, 2.2-2.4 (1H, 2.6-2.8 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.72 (1H, d, J=9.lHz), 8.0-8.2 (5H, 8.71 (1H, d, J=2.3Hz) APCI MASS (i/z)(Positive): 492.13 Preparation Methyl 4-[5-[6-[4-(4-ethylcyclohexyl)-l-piperazifl-l- 3 pyridylil-1, 3, 4-thiadiazol-2-yllbenzoate NMR (CDC1 3 0.8-2.0 (14H, in), 2.2-2.4 (1H, 2.6-2.8 (4H, 3.6-3.8 (4H, 3.96 (3H, 6.72 (1H, d, J=8.9Hz), 8.0-8.2 (5H, 8.71 (11H, d, J 2.4Hz) (+)APCI MASS (m/z)(Positive): 492.13 Preparation 56 To a mixture of ethyl 4-(piperazinyl)benzoate (2.00 g) and 4-methylenecyclohexan-l-oe (0.98 g) in a mixture of methanol ml) and acetic acid (1.47 ml) was portionwise added sodium cyanoborohydride (644 mg) with stirring under ice-cooling and WO 01160846 PCT/JPOI/01204 61 the mixture was stirred at ambient temperature overnight. To the reaction mixture was added water (200 ml) and the mixture was adjusted to pH 8-9 with saturated aqueous sodium hydrogen carbonate. The resulting precipitates were collected and chromatographed on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (2:1 v/v) The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl methylenecyclohexyl) -l-piperazinyllbenzoate (1.39 g).
NMR (CDCl 3 5) 1. 36 (3H, t, J=7.l12Hz) 1. 39-1.50 (2H, mn) 1.05-2.15 (4H, in), 2.30-2.60 (3H, mn), 2.65-2.75 (4H, in), 3. 25-3. 35 (4H, mn), 4. 32 (2H, q, J=7 .l2Hz), 4. 63 (2H, s) 6. 85 (2H, d, 08Hz) 7. 92 (2H, d, J=9 04Hz) EST MASS (Positive): 329.4 The following compounds [Preparations 57 to 63] were obtained according to a similar manner to that of Preparation 56.
Pre-paration 57 Ethyl 4- (4-phenylcyclohexyl) -l-piperazinyljbenzoate NMR (CDCl 3 8) 1. 36 (3H, t, J7.lHz) 1. 40-2.12 m) 2.31 (1H, br), 2.62-2.67 (4H, mn), 3.32-3.37 (4H, mn), 3.6-3.8 (1H, mn), 4.33 (2H, q, J=7.lHz), 6.87 (2H, di, J=r9Hz), 7.1-7.35 (5H, mn), 7.92 (2H, d, J=9Hz) APCI MASS (Positive) 393.33 Preparation 58 Ethyl 4- (cyclohexylinethyl) -1-piperazinyilbenzoate NMR (CDCl 3 5) 0. 75-1. 00 (2H, mn), 1. 36 (3H, t, J=7.l1Hz) 1-10-1.82 in), 2.17 (2H, di, J=7.lHz), 2.50-2.55 (4H, in), 3.29-3.34 (4H, mn), 4. 32 (2H, q, J=7.l1Hz) 6.85 (2H, di, J=9Hz), 7.92 (2H, di, JTh9Hz) APCI MASS (in/z) (Positive) 331.4 WO 01160846 PCT/JPO/01204 62 Preparation 59 Ethyl 4-[4-[4-(7-methoxyheptyoxy)cyclohexyl]-lpiperazinyl]benzoate IR (Neat): 1707, 1606, 1518, 1452, 1389, 1367, 1282, 1236, 1188, 1119, 1107 cnC' NMR (CDC1 3 1.2-2.2 (21H, 2.2-2.4 (1H, 2.7-2.8 (4H, mn), 3.33 (3H, 3.1-3.8 (9H, 4.32 (2H, q, J=7.lHz), 6.86 (2H, d, J=9.OHz), 7.92 (2H, d, J=9.OHz) AOCU MASS (m/z)(Positive): 461.53 Preparation Ethyl 4-[4-[4-(8-methoxyoctyloxy)cyclohexyl]-lpiperazinyl]benzoate IR (Neat): 2933, 2856, 1705, 1608, 1516, 1454, 1282, 1238, 1111 cin 1 NMR (CDC1 3 1.2-2.2 (23H, 2.2-2.4 (1H, 2.6-2.8 (4H, 3.1-3.5 (91H, 3.33 (3H, 4.32 (2H, q, J=7.11Hz), 6.86 (2H, d, J=9.0Hz), 7.92 (2H, d, ESI MASS (m/z)(Positive): 475.5 (M 1 +1) Preparation 61 Methyl 6-[4-(cis-4-methylcyclohexyl)-lpiperazinyl]nicotinate NMR (CDC1 3 0.94 (3H, d, J=6.9Hz), 1.40-1.82 (9H, 2.16-2.26 (lH, 2.59-2.65 (41, 3.65-3.71 (4H, 3.86 (3H, 6.58 (1H, d, J=9Hz), 8.00 (1H, dd, J=2.4 and 9Hz), 8.79 (1H, d, J=2.4Hz) APT-ES MASS (i/z)(Positive): 318.3 (Ml-1) Methyl 6-[4-(trans-4-iethylcyclohexyl)-lpiperazinyl]nicotinate NMR (CDC1 3 0.88 (3H, d, J=6.4Hz), 0.90-1.40 (4H, n), 1.70-1.95 (5H, 2.20-2.35 (1H, 2.63-2.68 (4H, 3.65-3.70 (4H, 3.86 (3H, 6.57 (1H, d, J=9Hz), 7.99 (1H, dd, J=2.4 and 9Hz), 8.78 (11, d, WO 01160846 PCT/JPOI/01204 63 J=2. .4Hz) API-ES MASS (rn/z) (Positive): 318.3 Pre-paration 62 Methyl 6-[4-(cis-4-ethylcYClohexyl)-l piperazinyll nicotinate PMR (CDCl 3 6) 0. 87 (3H, t, J=7. 3Hz) 1. 25-1. 67 (11H, in), 2.20-2.30 (1H, mn), 2.59-2.64 (4H, in), 3.65-3.70 (4H, in),r 3. 86 (3H, s) 6. 58 (1H, d, c=9.l1Hz) 8. 00 (iH, cid, J=2.4 and 9.1Hz), 8.79 d, J=2.4Hz) APT-ES MASS (Positive) 332.4 (M 4 +1) Methyl 6- (trans-4-ethylcyclohexyl) -1piperazinyllnicotinate NMR (CDC' 3 6) 0.087 (3H, t, J=7. 5Hz) 0. 91-1. 40 (5H, m) 1. 6-2. 0 (6H, mn), 2. 64-2. 69 (4H, m) 3. 66-3.71 (4H, m) 3. 86 (3H, s) 6. 57 (1H, d, J 9Hz) 3. 00 (TH, dci, J=2. 3 and 9Hz), 8.79 (1I, d, J=23Hz) API-ES MASS (rn/z) (Positive) 332.4 Prepoaration 63 4- (l-Cyclohexyl-4-piperidyl)beflzonitrile IP. (KBr) 2927, 2852, 2222, 1605, 1504, 1450 cmf
T
NMR (DMSO-d 6 8) 1. 0-2. 0 (14H, in), 2.2-2. 7 (4H, mn), 2.08-3. 1 (2H, in), 7. 45 (2H, ci, J=8. 3Hz) 7. 75 (2H, d, J=8. 3Hz) APCI MASS 269.33 Prepoaration 64 A mixture of 7-bromo-1-heptanol (25 g) and sodium inethoxide, 28% solution in methanol (37 ml) in methanol (250 ml) was stirred for 7 hours at 90'C. After being cooled to room temperature, the solvent was evaporated in vacun. The residue was purified by column chromatography on silica gel eluting with a mixture of dichlcromethane and methanol (100:1-,25:1). The eluted fractions containing the desired product were collected and WO 01/60846 PCT/JP01/01204 64 evaporated in vacuo to give 7-methoxy-l-heptanol (18.2 g).
NMR (CDC1 3 1.2-1.7 (10H, 3.33 (3H, 3.3-3.4 (2H, 3.5-3.7 (2H, m) The following compound was obtained according to a similar manner to that of Preparation 64.
Preparation 8-Methoxy-l-octanol NMR (CDC1 3 1.2-1.7 (12H, 3.33 (3H, 3.3-3.4 (2H, 3.5-3.7 (2H, m) Preparation 66 To a mixture of 7-methoxy-l-heptanol (18.1 g) and ptoluenesulfonyl chloride (28.4 g) in dichloromethane (180 ml) was added triethylamine. After stirring for 27.5 hours at room temperature, the solvent was evaporated in vacuo. Then the residue was poured into a mixture of ethyl acetate and water.
The organic layer was successively washed with water and brine and dried over magnesium sulfate. The solvent was evaporated in vacuo and the residue was purified by column chromatography on silica gel eluting with a mixture of hexane and ethyl acetate The eluted fractions containing the desired product were collected and evaporated in vacuo to give 7-methoxyheptyl 4-methylbenzenesulfonate (28.5 g).
NMR (CDC1 3 1.1-1.7 (10H, 2.45 (3H, 3.3-3.4 4.02 (2H, t, J=6.5Hz), 7.3-7.4 (2H, 7.7-7.8 (2H, m) ESI MASS (m/z)(Positive): 323.3 (M++Na) The following compound was obtained according to a similar manner to that of Preparation 66.
Preparation 67 8-Methoxyoctyl 4-methylbenzenesulfonate WO 01/60846 PCT/JP01/01204 NMR (CDC1 3 1.1-1.7 (12H, 2.45 (3H, 3.3-3.4 4.01 (2H, d, J=6.5Hz), 7.3-7.4 (2H, 7.7-7.9 (2H, m) ESI MASS (m/z)(Positive): 337.2 (M++Na) Preparation 68 A mixture of 8-(7-methoxyheptyloxy)-l, 4 (9.8 g) and 3N aqueous hydrochloric acid (34 ml) in tetrahydrofuran (68 ml) was stirred for 25.5 hours at room temperature. The solvent was evaporated in vacuo and the residue was poured into a mixture of ethyl acetate and water.
Then the solution was adjusted to pH 9 with potassium carbonate.
The organic layer was successively washed with water and brine and dried over magnesium sulfate. The solvent was evaporated in vacuo to give 4-(7-methoxyheptyloxy)cyclohexanone (8.34 g).
NMR (CDC1 3 1.2-2.4 (16H, m) 2.5-2.7 (2H, m) 3.33 (3H, 3.3-3.5 (4H, 3.6-3.8 (1H, m) ESI MASS (m/z)(Positive): 265.4 (M++Na) The following compound was obtained according to a similar manner to that of Preparation 68.
Preparation 69 4-(8-Methoxyoctyloxy)cyclohexanone NMR (CDC1 3 1.2-2.4 (18H, 2.5-2.7 (2H, 3.33 (3H, 3.3-3.5 (4H, 3.6-3.8 (1H, m) ESI MASS (m/z)(Positive): 279.3 (M++Na) Preparation To a solution of tert-butyl 4-[4'-(methoxycarbonyl)- 1,1'-biphenyl-4-yl]-l-piperazinecarboxylate (4.5 g) and anisole (6.17 ml) in dichloromethane (45 ml) was added dropwise with stirring trifluoroacetic acid (22.5 ml) at 0°C. The mixture was then stirred for 2 hours at room temperature. To the reaction mixture was added water. The resulting precipitate was collected WO 01160846 PCT/JPOI/01204 66 by filtration and washed with isopropanol and diisopropyl ether to give methyl (1-piperazinyl) -biphenyl-4-carboxylate trifluoroacetate (4.13 g).
NMR (CDC1 3 3.15-3. 55 (8H, mn), 3. 87 (3H, s) 7. 11 (2H, d, J=8.8Hz), 7.69 (2H, d, J=8.7Hz), 7.79 d, J=8.4Hlz), 8.00 (2H, d, J=8.4Hz), 8.84 (2H, br s) MASS (Positivc): 297 The following compounds [Preparation 71 to 731 were obtained according to a similar manner to that of Preparation Preparation 71 4- (5-Methoxypentyloxy) -1-piperidine trifluoroacetate This compound was immediately used as the starting compound for the next step.
Preparation 72 4- (6-Methoxyhexyloxy) piper idine This compound was immediately used as the starting compound for the next step- Preparation 73 4- (4-Piperidy1)benzonitrile IR (KBr) :2937, 2227, 1684, 1608, 1541, 1504, 1450, 1419, 1201, 1134, 1014, 835 cm'1 NMR (DMS0-cl 6 1.4-1.8 (4H, in), 2.5-2.8 (3H, in), 3.1 (2H, mn), 7.4-7.5 (2H, in), 7.7-7.8 (2H, mn) +)APCT MASS 187.27 Preparation 74 To an ice cooled solution of 1-methoxy-bicyc1ohexyli4-one (0.9 g) and methyl 4r-(l-piperazinyl)-l,1'hiphelyli 4 WO 01/60846 PCT/JP01/01204 67 carboxylate trifluoroacetate (2.11 g) in a mixed solvent of methanol (18 ml), tetrahydrofuran (14 ml) and acetic acid (0.735 ml) was added sodium cyanoborohydride (296 mg) in a stream of nitrogen. The mixture was stirred at room temperature for 5 hours.
The reaction mixture was quenched with saturated aqueous sodium hydrogen carbonate solution. The resulting precipitate collected by filtration, washed with water, isopropyl alcohol and diisopropyl ether, then dried to give methyl 4'-[4-[cisl-methoxy-l,1'-bi(cyclohexyl)-4-yl]-1-piperazinyl]-1,1'biphenyl-4-carboxylate and 4'-[4-[trans-l-methoxy-1,1'bi(cyclohexyl)-4-yl]-1-piperazinyl]-1,1'-biphenyl-4carboxylate (444 mg).
4'-[4-[Cis-l-methoxy-l,1'-bi(cyclohexyl)-4-yl]-1piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 0.8-1.9 (19H, 2.15-2.4 (1H, 2.7- 2.9 (4H, 3.11 (3H, 3.2-3.4 (4H, 3.93 (3H, 6.99 (2H, d, J=8.8Hz), 7.5-7.7 (4H, 8.06 (2H, d, MASS 491 (M +1) 4'-[4-[Trans-l-methoxy-1,1'-bi(cyclohexyl)-4-yl] -1piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 0.8-2.3 (20H, m) 2.6-2.75 (4H, m) 3.15 (3H, 3.2-3.4 (4H, 3.93 (3H, 7.00 (2H, d, J=8.8Hz), 7.5-7.7 (4H, 8.06 (2H, d, J=8.4Hz) MASS 491 The following compounds [Preparation 75 to 81] were obtained according to a similar manner to that of Preparation 74.
Preparation Benzyl 4-(trans-4-cyclohexylcyclohexyl)-1piperazinecarboxylate WO 01/60846 PCT/JPO/01204 68 IR (KBr): 1682, 1466, 1429, 1240 crn 1 NMR (CDC1 3 0.75-1.35 (12H, 1.5-1.95 (6H, n), 2.1-2.3 (1H, m) 2.4-2.6 (4H, 3.50 (4H, L, J 5.0Hz) 5.13 (2H, 7.25-7.4 (5H, m) EST MASS (r/z)(Positive): 385 (M+l) Preparation 76 Benzyl 4-(trans-4-tert-butylcyclohexyl)-lpiperazinecarboxylate IR (KBr): 1684, 1468, 1525, 1242 cm NMR (CDC1 3 6) 0.83 (9H, s) 0.9-1.58 (5H, 1.7-2.35 2.45-2.6 (4H, 3.51 (4H, t, J=5.lHz), 5.13 (2H, 7.35 (5H, s) MASS 359 (M+1I) Preparation 77 Methyl 4'-[4-(trans-4-ethylcyclohexyi)-l-piperazinyl]- 1,1'-biphenyl-4-carboxyiate NMR (CDC1 3 0.8-2.05 (14H, 2.2-2.4 (1H, 2.65-2.8 (4H, 3.2-3.35 (4H, 3.92 (3H, 6.99 (2H, d, J=8.8Hz), 7.55 (2H, d, J=8.8Hz), 7.62 (2H, d, J=8.4Hz), 8.06 (2H, d, J=8.4Hz) MASS 407 (M+1) Methyl 4'-[4-(cis-4-ethylcyclohexyl)-l-piperazill- 1,1'-biphenyl-4-carboxylate NMR (CDC1 3 0.88 (3H, t, J=7.2Hz), 1.2-1.7 (11H, m), 2.2-2.4 (1H, 2.65-2.8 (4H, 3.2-3.35 (4H, n), 3.93 7.00 (2H, d, J=8.8Hz), 7.56 (2H, d, J=8.8Hz), 7.62 (2H, d, J=8.4Hz), 8.06 (2H, d, J=8-AHz) MASS 407 Preparation 78 Methyl 4'-[4-[trans-4-(trans-4'-methoxycyclohexyl-l'- VI]cyclohexyl-1-yl -l-piperazinyl]-1,11-biphenyl-4- WO 01/60846 PCT/JPO/01204 69 carboxylate NMR (CDC1 3 0.8-1.4 (10H, mn), 1.65-2.4 (9H, in), 2.65-2.8 (4H, 2.95-3.15 (1H, 3.2-3.35 (4H, 3.35 (3H, 3.93 (3H, 6.99 (2H, d, J=8.8Hz), 7.55 (2H, d, Jh8.8Hz), 7.62 (2H, d, J=8.4Hz), 8.06 (2H, d, J=8.4Hz) MASS 491 (M+l) Methyl 4'-[4-[cis-4-(trans-4'-iethoxycyclohexyl-l' yl)cyclohexyl-1-yll-l-piperazinyll-1,1'-biphenyl-4carboxylate NMR (CDCl 3 0.8-2.3 (19H, 2.6-2.75 (4H, 2.95-3.2 (1H, in), 3.2-3.35 (4H, 3.35 (3H, 3.93 (3H, s), 7.00 (2H, d, J=8.SHz), 7.56 (2H, d, J=8.8Hz), 7.62 (2H, d, J=8.4Hz), 8.06 (2H, d, J8.4Hz) MASS 491 (M+l) Preparation 79 Methyl 4'-[4-[4-(trans-4-iethoxyphenyi)cyclohexY1]-1piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 1.4-1.7 (4H, 1.9-2.15 (4H, 2.3-2.6 (2H, n, 2.75-2.85 (4H, 3.2-3.4 (4H, 3.79 (3H, 3.93 (3H, 6.84 (2H, d, J=8.6Hz), 7.01 (2H, d, J=8.8Hz), 7.14 (2H, d, J=8.6Hz), 7.56 (2H, d, J=8.8Hz), 7.63 (2H, d, J=8.4Hz), 7.06 (2H, d, J=8.4Hz) MASS 485 (Ms+1) Methyl 4'-[4-[4-(cis-4-iethoxyphenyl)cyclohexyl-lpiperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 1.5-2.6 (10H,m), 2.6-2.75 (4H, 3.2-3.4 (4H, 3.79 (3H, 3.93 (3H, 6.8-6.9 (2H, m), 7.01 (2H, d, J=8.8Hz), 7.19 (2H, d, J=8.6Hz), 7.65 (4H, 8.06 (2H, d, MASS 485 (M+1) Preparation WO 01/60846 PCT/JP01/01204 Methyl 4'-[4-[cis-4-methoxy-(4-cyclopentyl)cyclohexyll-yl]-1-piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 1.2-1.9 (16H, 2.05-2.4 (2H, 2.7- 2.85 (4H, 3.16 (3H, 3.25-3.35 (4H, 3.93 (3H, 7.00 (2H, d, J=8.8Hz), 7.5-7.7 (4H, m), 8.0-8.1 (2H, m) MASS 477 Methyl 4'-[4-[trans-4-methoxy-(4-cyclopentyl)cyclohexyl-l-yl]-1-piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC1 3 1.35-1.9 (16H, 2.2-2.35 (2H, m), 2.6-3.75 (4H, 3.19 (3H, 3.2-3.35 (4H, 3.93 (3H, 7.00 (2H, d, J=8.9Hz), 7.56 (2H, d, J=8.9Hz), 7.62 (2H, d, J=8.6Hz), 8.06 (2H, d, J=8.6Hz) MASS 477 Preparation 81 Methyl 4'-[4-(cis-4-methoxy-4-phenylcyclohexyl)-1piperazinyl]-1,1'-biphenyl-4-carboxylate NMR (CDC13, 1.5-2.0 (6H, 2.1-2.6 (3H, 2.75-2.9 (4H, 2.99 (3H, 3.25-3.4 (4H, 3.93 (3H, s), 7.01 (2H, d, J=8.8Hz), 7.2-7.5 (5H, 7.57 (2H, d, J=8.8Hz), 7.63 (2H, d, J=8.5Hz), 8.06 (2H, d, MASS 485 Preparation 82 A mixture of methyl 4'-[4-[l-methoxy-l,1'bi(cyclohexyl)-4-yl]-1-piperazinyl]-1,1'-biphenyl- 4 carboxylate (440 mg) and 10% sodium hydroxide solution (1.4 ml) in a mixed solvent of methanol (8 ml) and tetrahydrofuran (24 ml) was refluxed for 4 hours. After cooling to ambient temperature, the reaction mixture was poured into cold water and the mixture was adjusted to pH 7 with 1.0 mol/1 hydrochloric acid.
The resulting precipitates were filtered, washed with water, isopropyl alcohol land diisopropyl ether, then dried to give WO 01160846 PCT/JPO/01204 71 4'-[4-[cis-l-methoxy-1,1'-bi(cyclohexyl) 4 -yll-lpiperazinyll-1,1'-biphenyl-4-carboxylic acid (371 mg).
MASS 477 (M+1) The following compounds [Preparations 83 to 1111 were obtained according to a similar manner to that of Preparation 82.
Preparation 83 4-[5-[4-[4-(Cyclohexylmethyl)-l-piperazinyllphenyl]- 1,3,4-thiadiazol-2-yllbenzoic acid NMR (DMSO-d 6 0.8-1.8 (11H, 2.5-3.5 (10H, m), 7.1-7.2 (2H, 7.92 (2H, d, 8.12 (4H, s) API-ES MASS 463.4 (M+1) Preparation 84 4-[5-[4-[4-[4-(7-Methoxyheptyloxy)cyclohexyl]-1piperazinyl]phenylhl-,3,4-thiadiazol-2-yl]benzoic acid NM (DMSO-d 6 6) 1.00-2.2 (19H, 2.8-3.6 (13H), 3.20 (3H, 7.14-7.18 (2H, 7.90-7.93 (2H, 8.03-8.22 (4H, m) ESI MASS (m/z)(Positive): 593.4 (M+1) Preparation 4-[5-[4-[4-[4-(8-Methoxyoctyl)cyclohexyll-lpiperazinyl]phenyl]-1,3,4-thiadiazol-2-yllbenzoic acid NMR (DMSO-d 6 1.1-2.2 (21, 3.0-3.6 (13H, 3.20 (3H, 7.14-7.18 (2H, 7.78-8.21 (6H, m) ESI MASS (m/z)(Negative): 607 (Mt+l) Preparation 86 4-[5-[4-[4-(5-MethoxypentyloXy)-l-piperidyllphenyl]- 1,3,4-thiadiazol-2-yl]benzoic acid hydrochloride NMR (DMSO-d 6 1.25-1.60 (8H, 1.80-1.95 (2H, m), 3.00-3.20 (2H, 3.21 (3H, 3.25-3.55 (5H, m), WO 01160846 PCT/JPOI/01204 72 3.60-3. 80 (2H, mn), 7. 08 (2H, di, J=8. 96Hz) 7. 84 (2H, d, J=8.48Hz), 8.10 (4H, s) ESI MASS (in/z) (Negative) 480.2 Preparati n 37 4- (6-Methoxyhexyloxy) -1-piperidyllphenyll 1,3, 4-thiadiazolL-2-ylilbenzoic acid NMR (DMSO-d 6 8) 1. 2-1. 6 (10H, mn), 1. 8-2. 0 (2H, in) 8 (12H, mn), 7. 08 (2H, d, J=9 -OHz) 7. 84 (2H, d, J=8 .8Hz), 8.11 (4H, br s) ()APCI MASS (Positive): 496.27 Preparation 88 4- (4-MethoxybutoxynethyJ2 -l-pip eridyliphenyl] 1,3, 4-thiadiazol-2-ylllbenzoic acid NMR (DMSO-d 6 1.2-1.9 (9H, in), 2.8-3.0 (2H, in), 3.2- (9H, in), 3.8-4.0 (2H, in), 7.07 (2H, d, J=8.9Hz), 7.84 (2H, d, J=8.7Hz), 8.10 (4H, br s) ()APCI MASS (in/z) (Positive): 482.20 Preparation 89 4- (5-Methoxypentyloxynethyl) -1piperidyllphenyll-1,3,4-thiadiazol-2-ylbelzoic acid NMR (DMSO-d 6 1.1-1.8 (11H,mi), 2.7-2.9 (2H,mi), 3.2-3.5 (9H, mn), 3.8-4.0 (2H, in), 7.07 (2H, d, J=9.OHz), 7.84 (2H, d, J=8.8Hz), 8.0-8.2 (4H, mn) ESI MASS (Negative) 494.3 Pregaration 4-[5-[6-[Cis-4-(4-rnethylcyclohexyl)-l-piperaziflyl-3pyridyl]-i, 3,4-thiadiazol-2-yllbelzoic acid APCT MASS (Positive) 464.13 Preparation 91 4-[5-[6-[Trans-4-(4-methylcyclohexyl)-l-piperazinll- 3 WO 01160846 PCT/JPOI/01204 73 pyridyl] 4-thiadiazol-2-yl]benzoic acid APCI MASS Positive): 464.20 (M Preparation 92 4-[5-[6-[Cis-4-(4-ethylcyclohexyl)-1-piperazill-3pyridyl] 4-thiadiazol-2-ylilbenzoic acid ESI MASS (Positive)- 478.3 Preparation 93 4-[5-[6-[Trans-4-(4-ethylcyclohexyl)-l-piPerazil]-3pyridyll 4-thiadiazol-2-yl] beuzoic acid APCI MASS (1n/z) (Positive): 4-78.3 Preparation 94 4-2[-ICcoey--ieiy~hnliiao21 b] [1,3,41]thiadiazo1-6-yl1benzoic acid hydrochloride IR (KBr): 2937, 1699, 1608, 1471, 1414, 1373, 1255, 1174 cm- 1 NMR (DIASO-d 6 1.0-2.1 (14H, mn), 2.8-4.0 7.4- (8H, in), 8.86 (1H, s) (IARCI MASS 487.33 Preparation 4- [2-114- [Cis-4- (4-methylcyclohexyl) -1piperazinyllphenyl]imidazo[2,1-b1[1,3,4]thiadiazol-6yllbenzoic acid hydrochloride ESI MASS (Positive): 502.3 Preiparation 96 [4-(Trans-4-cyclohexylcyclohexyl) -1-piperazinyl] 1,1' -biphenyl-4-carboxylic acid ESI MASS 447 Preparation 97 WO 01/60846 PCT/JPOI/01204 74 (Trans-4-tert-butylcyclohexyl) -1-piperazinyl- 1,1' -biphenyl-4-carboxylic acid ESI MASS 421 Preparation 98 (Trans-4-ethylcyclohexyl) -1-piperazinyll biphenyl-4--carboxylic acid TR (KBr): 1699, 1602, 1525, 1377 cmC 1 MASS :393 Preparation 99 (Cis-4-ethylcyciohexyl) -1-piperazinyl] biphenyl-4-carboxylic acid IR (KBr): 1691, 1603, 1529, 1452, 1381 cm7 1 MASS 393 Preparation 100 4'-[4-[Tran3-1-rnethoxy-1,1'-bi(cyclohexyl)- 4 -yllpiperazinyl] -biphenyl--4-carboxylic acid MASS 477 Preparation 101 4' (Cis-4- (trans-4-methoxycyclohexyl-1-yl) cyclohexyl-1-yl) -'-piperazinyl] -biphenyl-4-carboxylic acid MASS 477 Preparation 102 [Trans-4- (cis-4-methoxycyclohexyl-ly1)cyclohexyl-1-yll-lpiperazilyl]1lrbiphel- 4 carboxylic acid MASS :477 Preparation 103 4'-[4-[Cis-4-(4-methoxyphenyl)cyclohexyll WO 01160846 PCT/JPOI/01204 piperazinyl] -biphenyl-4-carboxylic acid MASS (mhz): 471 Preparation 104 (4-Methoxyphenyl)-1-piperazil]-1,l'-bipheli- 4 carboxylic acid MASS 491 Preparation 105 4'-[4-[Trans-4-(4-methoxyphenlycYCohexyl1-lpiperazinyll-1,1' -biphenyl--4--carboxylic acid MASS 471 Preparation 106 4'[-44Dmtyccoey)lpprzni-,' biphenyl-4-carboxylic acid MASS 393 Pre-paration 107 41-[4[Cis4methoxy-(4-cyclopeltyl)cyclohexyl-livll piperazinyl] -biphenyl-4-carboxyiic acid MASS 463 Preparation 108 41[-Tas4mtoy(-ylpny~ylhxl1yl I-piperaziflyll1,1'-biphenyl-4-carboxylic acid MASS :461 Prenaration 109 (Cis-4-methoxy-4-phelylcyclohexyl) -1piperazinyl] -biphenyl-4-carboxyJ-ic acid' MASS 471 Preparation 110 4-5[-4(-ehlncclhxl--ieaiyl WO 01/60846 PCT/JP01/01204 76 phenyl]-1,3,4-thiadiazol-2-yl]benzoic acid dihydrochloride IR (KBr): 3400, 2939, 2852, 2592, 2455, 1705, 1603, 1522 cm- 1 NMR (DMSO-d 6 1.20-1.80 (4H, 1.80-2.40 (4H, m), 2.50-2.60 (4H, 3.10-3.30 (4H, 4.70 (1H, br s), 7.16 (2H, d, J=8.6hz), 7.92 (2H, d, J=9.2Hz), 8.00-8.30 (4H, m) API-ES MASS (m/z)(Positive): 463 (M+-2HC1+1) Preparation 111 4-[5-[4-[4-(4-Phenylcyclohexyl)-1-piperazinyl]phenyl]-1,3,4-thiadiazol-2-yl]benzoic acid API-ES MASS 525.3 (Ml+1) Preparation 112 A mixture of 4'-[cis-4-[l-methoxy-l,1'-bi(cyclohexyl)- 4-yl]-1-piperazinyl]-1,1'-biphenyl-4-carboxylic acid (367 mg), l-hydroxybenzotriazole (208 mg), 1-ethyl-3-(3'dimethylaminopropyl)carbodiimide hydrochloride (443 mg) and triethylamine (0.216 ml) in methylene chloride (37 ml) was stirred for 23.5 hours at room temperature then evaporated under reduced pressure. Water was added to the residue and the resulting precipitate collected by filtration, washed with water, isopropyl alchol and diisopropyl ether, then dried to give 1-[[4'-[cis-4-[1-methoxy-l,1'-bi(cyclohexyl)-4-yl]-lpiperazinyl]-1,1'-biphenyl-4-yl]carbonyloxy]-IH-1,2,3benzotriazole (399 mg).
NMR (CDC1 3 0.8-2.2 (19H, 2.3-2.5 (1H, 2.75- 2.95 (4H, 3.11 (3H, 3.25-3.5 (4H, 7.03 (2H, d, J=8.8Hz), 7.4-7.7 (5H, 7.79 (2H, d, 8.12 (1H, d, J=8.1Hz), 8.30 (2H, d, MASS 594 (M+1l) The following compounds [Preparations 113 to 138] were obtained according to a similar manner to that of Preparation WO 01/60846 PCT/JPO/01204 77 112.
Preparation 113 1-[4-[5-[4-[4-(Cyclohexylethyl)-l-piperazinyliphenyll- 1,3,4-thiadiazol-2-yl benzoyoxy--1H-1,2,3-benz otriazole TR (KBr): 2922, 2845, 1780, 1603, 1441, 1416, 1232, 984, 822 cm-1 NMR (CDC1 3 0.8-1.9 (11H, 2.22 (2H, d, J=7.1Hz), 2.5-2.7 (4H, 3.3-3.5 (4H, 6.9-8.5 (12H, m) APCI MASS 580.13 (M+1) Preparation 114 1-[4-[5-[4-[4-[4-(7-Methoxyheptyloxy)cyclohexyl]-1piperazinyl]phenyl]-1,3,4-thiadiazol-2-yllbenzoyloxy]-1H- 1,2,3-benzotriazole IR 2931, 2856, 1778, 1603, 1441, 1416, 1234, 1093, 984 cm 1 NMR (CDC1 3 6) 1-0-2.2 (19H, 2.4-3.7 (13H, 3.33 (3H, 6.8-8.5 (12H, m) Preparation 115 1-[4-[5-[4-[4-[4-(8-Methoxyoctyloxy)cyclohexyll-1piperazinyl]phenyl]-1,3,4-thiadiazol-2-ylibenzoyloxy]-1H- 1,2,3-benzotriazole IR CKBr): 2931, 2856, 1778, 1605, 1441, 1416, 1234, 1093, 984 cmf 1 NMR (CDC1 3 6) 1.0-2.2 (21H, 2.7-3.7 (13H, mn), 3.33 (3H, 6.9-8.5 (12, m) Preparation 116 1-[4-[5-[4-[4-(5-Methoxypentyloxy)-l-pipericyllphenylj- 1,3,4-thiadiazol-2-ylbenzoyloxyl-1H-1,2,3-benzotriazole NMR (DMSO-d 6 1.30-1.80 (81, 1.85-2.1C (2H, m), 3.00-3.25 (2H, 3.34 (3H, 3.35-3.55 (5H, m), 3.60-3.80 (2H, 6.97 (2H, d, J=8.95Hz), 7.35-7.65 WO 01/60846 PCT/JPO/01204 78 (3H, 7.90 (2H, d, J=8.81Hz), 8.13 (2H, d, J=8.19Hz) 8.23 (2H, d, J=8.46Hz), 8.39 (2H, d, J=8.41Hz) Preparation 117 1-[4-[5-[4-[4-(-Methoxyhexyloxy)-l-pipe 1,3,4-thiadiazol-2-yl]benzoyloxyl-TH-1,2,3-benzotriazole TR (KBr): 2931, 2856, 1778, 1603, 1439, 1416, 1230, 1109, 982 cnJ' NMR (CDC1 3 1.3-2.1 (12H, 3.0-3.3 (2H, 3.33 (3H, 3.3-3.6 (5H, 3.6-3.8 (2H, 6.97 (2H, d, J=8.9Hz), 7.4-7.7 (3H, 7.90 (2H, d, J=8.8Hz), 8.1-8.3 (3H, 8.3-8.5 (2H, m) APCI MASS (m/z)(Positive): 612.93 (M +1) Preparation 118 1-[4-[5-[4-[4-(4-Methoxybutoxyethyl)-lpiperidyl]phenyll-1,3,4-thiaciazol-2-yl]benzoylY-1H- 1,2,3-benzotriazolc IR (KBr): 1778, 1603, 1439, 1412, 1248, 1230, 1115, 1090, 984 cirf NMR (DMSO-d 6 1.0-2.1 (9H, 2.8-3..0 (2H, 3.1- (6H, 3.34 (3H, 3.8-4.0 (2H, 6.96 (2H, d, J=9.OHz), 7.3-7.6 (3H, 7.90 (2H, d, J=8.9z), 8.12 (1H, d, J=7.2Hz), 8.2-8.3 (21, 8.3-8.5 (2H, n) Preparation 119 1-[4-[5-[4-[4-(5-Methoxypentyloxymethyl)ll piperidyl]phenyl]-1,3,4-thiadiazol-2-ylbenzoyloxy]lH 1!2,3-benzotriazole NMR (CDC1 3 1.1-2.0 (1111, in), 2.7-2.9 (2H, 3.2-3.5 (6H, 3.33 (3H, 3.7-3.9 (2H, 6.90 (2H, m, J=9.OHz), 7.3-7.6 (3H, 7.83 (2H, d, J=8.8Hz), 8.06 (1H, d, J=8.21z), 8.16 (2H, d, J=8.5Hz), 8.33 (2H, d, WO 01160846 PCT/JPO/01204 79 H) APCI MASS (i/z)(Positive): 613.13 (M+l) Preparation 120 1-[4-[5-[6-[4-(4-Methylcyclohexyi)-l-piperazifll- 3 pyridyl]-1,3,4-thiadiazol-2-yl]benzoyloxy]-1H-1,2, 3 benzotriazole IR (KBr): 2947, 2922, 1778, 1601, 1429, 1400, 1236, 987 cm- 1 NMR (CDC1 3 0.8-1.5 (8H, 1.7-1.9 (2H, 1.9-2.1 (2H, 2.6-3.0 (1H, 3.0-3.2 (4H, br 4.2 (4H, br s) 6.6-6.8 (1H, 7.4-8.5 (9H, mi), 8.75 (1H, d, J=2.3Hz) ESI MASS (m/z)(Positive): 581.3 Preparation 121 1-[4-[5-[6-[4-(4-Ethylcyclohexyl)-l-piperazifll-3pyridyl]-1,3,4-thiadiazol-2-yllbenzoyloxyl--1,2,3benzotriazole IR (KBr) 2926, 1780, 1703, 1601, 1508, 1429, 1402, 1379, 1242, 984 crn 1 NMR (CDC1 3 0.8-1.0 (5H, 1.2-1.9 (8H, 1.9-2.5 (2H, 2.6-2.8 (4H, 3.6-3.8 (4H, 6.73 (1H, d, J=9.lHz), 7.4-7.7 (3H, 8.0-8.5 (6H, 8.74 (1H, d, J=2.3Hz) EST MASS (n/z)(Positive): 595.3 (Mf+1) Preparation 122 1-[4-[5-[6-[Trans-4-(4-ethylcyclohexyl)-1-piperazinyll- 3-pyridyll-1,3,4-thiadiazol-2-yl]benzoyloxY]-TH-1,2,3benzotriazole IR (KBr) 2924, 2850, 1778, 1601, 1429, 1402, 1362, 1244, 984 cmr 1 NMR (CDC1 3 0.8-1.5 (10H, 1.7-2.1 (4H, mi), 2.3-2.6 (1H, 2.7-2.9 (4H, br 3.6-3.9 (4H, br s), 6.6-6.8 (1H, 74-7.7 7.9-8.5 (6H, 8.74 (1H, d, J=2.3Hz) WO 01/60846 PCT/JPOI/01204 ESI MASS (Positive): 595.3 (M Preparation 123 1-[4-[2-[4-[Cis-4-(4-methylcyclohexyi)-1piperazinyl]phenyl]imidazo[2,1-b] [1,3,4jthiadiazol-6ylllbenzoyloxyl -1H-1,2, 3-benzotriaz'ole TR (KBr) :1795, 1697, 1649, 1605, 1539, 1473 1383, 1234, 1095, 1018 cm1 Preparation 124 1- (Trans-4-cyclohexyicyclohexyl) -1-piperazinyl] 1,1' -bipheny1-4-y1l carbonyloxy-lH-1, 2, 3-benzotriazoie NMR (CDC 1 3 8) 0. 8 5 (2 1H, mn), 2.08- 3. 0 (4 H, mr) 3. 3-3.4 (4KH, in), 7. 03 (2 H, d, J= 8- 7 Hz) 7. 4 7 (5 H, nm), 7.7 9 (2H, d, J=8.4H-z), 8.12 (1H, d, J=8.4Hz),8.30 (2H, d, J=8 .3Hz) MASS :447 Preparation 125 1-[4--(5-[4-[4-(4-Methylenecyclohexyl)-1piperazinyl] phenyl] 4-thiadiazol-2-yl] benzoyloxy] -1H- 1,2, 3-benzotriazole IR 3425, 3404, 2929, 2831, 1780, 1600 cm- 1 NMR (DMSO-d 6 1.10-1.50 (4H, mn), 1.60-2.20 (4H, in), 2. 20-2. 40 (3H, in), 2. 50-2. 80 (4H, mn), 4. 62 (1H, br s) 7-17 (2H, d, J=9.lHz), 7.70-7.60 (2H, in), 7.65 (1K, d, 7.75-8.00 (3H, mn), 8.10-8.30 (4H, in) API-ES MASS (Positive) 584 Preparation 126 1-[4'-[4-(Trans-4-tert-butylcyclohexyl)-1-piperaziflyl]- 1,1'-biphenyl-4-yllcarbonyioxy-lH-1;'2,3-benzotriazole IR (KBr): 1770, 1570, 1236 c- NMR (CDC1 3 8) 0. 86 (9H, s) 0. 9-1. 4 (5H, mn), 1. 7-2. 5 mn), 2.7-2.85 (4H, mn), 3.2-3.4 (4H, mn), 6.9-7.1 (2H, WO 01/60846 PCT/JPOI/01204 81 in), 7.3-7.7 (5H, mn), 7.79 (2H, di, J=8.5Hz) 8.11 (1H-, d, J=8.4Hz), 8.30 (2H, di, J=8.4Hz) MASS (in/z) 538 Preparation 127 1- (Trans-4-ethylcyclohexyl) -1-piperazinyl] biphenyl-4-yl] carbonyloxy-1H-1, 2, 3-benzotriazole Preparation 128 1-[4'-[4-(Cis-4-ethylcyclohexyl)-1.-piperazinyll-1,1'biphenyl-4-yl] carbonyloxy-1H-1, 2, 3-benzotriazole Pre-paration 129 1- [Trans-l-methoxy-1, 1' -bi (cyclohexyl) -4-yl] -1piperazinyl] -biphenyl-4-yl] carbonyloxy-TE-1,2, 3benzotriazole NMR (CDC1 3 8-2. 6 (20R, mn), 2. 7-3. 0 (4H, in), 3. 15 (3H, 3.3-3.6 in), 7.03 (2H, d, J=8.7Hz), 7-3-7.7 mn), 7. 79 (2H, di, J=8. 4H-z) 8. 12 (1H, di, J=8. lHz), 8.31 (2H, d, J~=8.4Hz) MASS 594 Preparation 130 1- [CiLs-4- (cis-4' -iethoxycyclohexy1-1' yl)cyclohexyl-1-yl]-1-piperazinyl]-1,1'-biphenyl-4yl] carbonyloxy-1H-1, 2, 3-benzotriazole NMR (CDC1 3 6) 0. 7-2. 6 (19H, mn), 2. 7-3. 2 (5H, in), 3. 35 (3H, 3.3-3.5 (4H, in), 7.03 (2H, d, J=8.8Hz), 7.35-7.7 in), 7. 79 d, J=8. SHz) 0. 12 (lE, d, J=8. 2Hz) 8.30 (2H, di, MASS 594 Preiparation 131 1- [Trans-4- (cis-4' -iethoxycyclohexyl-1'y1)cyclohexy1-1-y1i-l-piperaziny1]-1,1'-bipheny1-4- WO 01/60846 PCT/JPO/01204 82 yllcarbonyloxy-1H- ,2,3-benzotriazole NMR (CDC1 3 0.8-1.5 (10H, 1.6-2.6 (9H, 2.8-3.2 3.35 (3H, 3.3-3.5 (4H, 7.03 (2B, d, J=8.8Hz), 7.4-7.7 (5H, 7.78 (2H, d, J=8.5Hz), 8.12 (1H, d, J=8.2Hz), 8.30 (2H, d, MASS 594 (MS+1) Preparation 132 1-[4'-[4-[Cis-4-(4-rethoxyphenyl)cyclohexyl]-1piperazinyll-1,1'-biphenyl-4-yl carbonyloxy-T-1,2,3benzotriazole NMR (CDC1 3 1.5-2.9 (14H, 3.3-3.5 (4H, mn), 3.79 (3H, 6.85 (2H, d, J=8.7Hz), 7.03 (2H, d, J=8.BHz), 7.21 (2H, d, J=8.7Hz), 7.4-7.7 (5H, 7.79 (2H, d, J=8.5Hz), 8.12 (1H, d, J=8.2Hz), 8.30 (2H, d, MASS 588 Preoaration 133 1-[4'-[4-(4-Methoxyphenyl)-1-piperazinyl]-1,1'biphenyl-4-yi] carbonyloxy-TH-1, 2, 3-benzotriazole NMR (CDC1 3 3.2-3.55 (811, 3.79 (311, 6.8-7.2 (6H, 7.4-7.9 (7H, 8.0-8.15 8.31 (1H, d, J=8.2Hz) MASS 506 (M+1) Preparation 134 1-[4'-[4-[4-(Trans-4-methoxyphenyl)cyclohexyl] piperazinyll-1,1'-biphenyl-4-yllcarbonyloxy-1H-1,2,3benzotriazole NMR (CDC1 3 1.4-2.35 (8H, 2.3-2.5 (2H, 2.8-3.0 (4H, 3.3-3.5 (4H, 3.79 (3H, 6.85 (2H, d, J=8.7 7z), 7.04 (2H, d, J=8.8Hz), 7.13 (2H, d, J8.7Hz), 7.4-7.7 7.79 (2H, d, J=0.5Hz), 0.12 (11, d, J=8.2Hz), 8.30 (2H, d, MASS 588 (Mt+1) WO 01/60846 PCT/JPOI/01204 83 Preparation 135 J-[4'-[4-(4,4-Dimethylcyclohexyl)-1-piperazinyl]-1,1'biphenyi-4-yl] carbonyloxy-1H-1, 2, 3-benzotriazole NMR (CDC1 3 8) 0. 92 s) 1.12-1. 9 (8H,m) 2.2-2.5 (1H, in), 2.75-2-95 (4H, mn), 3.3-3.45 (4H, mn), 7.03 (2H, d, J=8.8Hz), 7.4-7.7 (5H, in), 7.79 (2H, d, J=8.5Hz), 8.12 (1H, d, J=8.2Hz), 8.30 (2H, d, MASS 510 (M+1) Preparation 136 1- [Cis-4-inethoxy- (4-cyclopentyl) cyclohexyl-1ylj -1-piperazinyl] -biphenyl-4-yl] carbonyloxy-TH-1, 2,3berizotriazole IR (KBr): 1776, 1597 cmn' NMR (CDC1 3 1.2-2.0 (iGH,mi), 2.1-2.45 (2H,mi), 2.75-2.9 (4H, mn), 3.16 (3H, 3.25-3-4 (4H, mn), 7.03 (2H, d, J=8.9Hz), 7.4-7.7 (5H, in), 7.79 (2H, d, J=8.6Hz), 8.11 (111, d, J=8.2Hz), 8.30 (2H, d, J=8.6Hz) MASS 579 Preparation 137 1- [Trans-4-inethoxy- (4-cyclopentyl) cyclohexyl-1yl] -1-piperazinyll carbonyloxy-1H-2,2, 3-benzotriazole IR (KBr): 1772, 1597 c- NMR (CDC1 3 1.35-2.4 (28H, in), 2.65-2.8 (4H, rn), 3.20 (3H, 3.25-3.4 (4H, in), 7.03 (2H, d, J=8.9Hz), 7.4-7.7 in), 7.79 (2H, d, J=8.6Hz), 8.05-8.15 (1H, in), 8.25-8.35 (2H, in) MASS 580 Preparation 138 1- (Cis-4-methoxy-4-phenylcyclohexyl) -1piperazinyl] -biphenyl-4-yl] carbonyloxy-1H-1, 2,3benzotriazole WO 01/60846 PCT/JP01/01204 84 NMR (CDC1 3 1.5-2.6 2.8-2.95 (4H, 2.99 (3H, 3.3-3.45 (4H, 7.05 (2H, d, J=8.9Hz), 7.2-7.7 7.79 (2H, d, J=8.6Hz), 8.11 (1H, d, J=8.1Hz), 8.30 (2H, d, J=8.6Hz) MASS 588 Preparation 139 Amixture of cesium trichloride (24.9 g) in tetrahydrofuran ml) was stirred at room temperature for 5 hours. 1,4- Dioxa-spiro[4.5]decan-8-one (1.4 g) was added to the solution and stirred at room temperature for 1 hour. To the solution was added dropwise with stirring phenylmagnesium chloride solution in dimethyl ether) (33.7 ml) at 0°C. The reaction mixture was quenched with 10% acetic acid aqueous solution.
Dimethyl ether was added to the solution. The organic layer was washed with brine, sodium hydrogen carbonate solution and brine and dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (3:1 hexane-ethyl acetate elution) to give 8-phenyl-l,4dioxaspiro[4.5]decan-8-ol (5.94 g).
NMR (CDC1 3 1.65-2.3 (8H, 3.99 (4H, 4.03 (1H, 7.2-7.6 (5H, m) MASS 257 (M+23) The following compound was obtained according to a similar manner to that of Preparation 139.
Preparation 140 8-Cyclopentyl-l,4-dioxaspiro[4.5]decan-8-ol NMR (CDC13, 1.2-2.1 (17H, 3.9-4.05 (4H, 4.03 (1H, s) MASS 249 (M++23) Preparation 141 WO 01/60846 PCT/JP01/01204 To a solution of 8-phenyl-1,4-dioxaspiro[4.5]decan-8-ol (5.76 g) and iodomethane (4.59 ml) in N,N-dimethylformamide (58 ml) was added sodium hydride (60% dispersion in mineral oil) (1.97 g) at 0 C. The solution was stirred for 2 hours at 0°C and at room temperature for 7.5 hours. The reaction mixture was added to a mixture of water and ether. The organic layer was washed with brine and dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (5:1 hexane-ethyl acetate elution) to give 8-methoxy-8-phenyl-l,4-dioxaspiro[4.5]decane (5.968 g).
NMR (CDC1 3 1.6-2.15 3.00 (3H, 3.9-4.05 (4H, 7.2-7.5 (5H, m) The following compounds [Preparations 142 and 143] were obtained according to a similar manner to that of Preparation 141.
Preparation 142 4'-Methoxy-1,1'-bi(cyclohexyl)-4-ol NMR (CDC13, 0.8-2.2 (18H, 2.9-3.6 (6H, m) MASS 235 (M++23) Preparation 143 8-Cyclopentyl-8-methoxy-l,4-dioxaspiro[4.5]decane NMR (CDC1 3 1.25-2.35 (17H, 3.16 (3H, 3.9-4.0 (4H, m) Preparation 144 A solution of 8-methoxy-8-phenyl-l,4- (5.96 g) and 3N-hydrrochloric acid (24 ml) in tetrahydrofuran was stirred at room temperature for 24 hours.
The reaction mixture was added to a mixture of sodium hydrogen carbonate solution and dimethyl ether. The organic layer was WO 01/60846 PCT/JPOI/01204 86 washed with sodium hydrogen carbonate solution and dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (1:0-150:1 dichioromethane-methanol elution) to give 4-methoxy-4phenylcyclohexanone (3.6 g).
NMR (CDCl 3 0-2. 9 (8H, mn), 3.-09 (3H, s) 7.25-7. 5
M)
MASS 227 (M++23) The following compound was obtained according to a similar manner to that of Preparation 144.
Preparation 145 4-Cyclopentyl-4-methoxycyciohexanone NMR (CDC1 3 8) 1.2-2.4 (15H, mn), 2.45-2.7 (2H, in), 3.27 (3H,, s) MASS 219 (M++l23) Preparation 146 A mixture of 4-[5-[4-[4-(4-phenylcyclohexyl)-lpiperazinyllphenyl]-l,3,4-thiadiazol-2-yl]benzoic acid (0.81 0-benzotriazol-1-yl-N,N,N' ,N'-tetramethyiuroniun hexafluorophosphate (0.66 g) and N,N-diisopropylethylamine (0.51 ml) in l-methyl-2-pyrrolidinone (16 ml) was stirred for 2 hours at 5000. The reaction mixture was poured into water.
Then the resulting precipitate was collected by filtration and washed with water to give 1-4-[5-[4-[4-(4-phenylcyclohexyl)- 1-piperazinyllphenyl]-1,3,4-thiadiazol-2-yl]benzoyloxy-lH- 1,2,3-benzotriazole (0.97 g).
IR (K<Br) :1780, 1603, 1444, 1414, 1234i 1100, 900, 043 cm 1 NMR (CDC1 3 1.6-2.2 (8H, mn), 2.3-2.5 (1H, mn), 2.7-2.9 mn), 3.4-3.6 (4H, mn), 6.9-8.5 (17H, m) AECI MASS 642.07 (MH-+l) WO 01/60846 PCT/JPOI/01204 87 The following compounds [Preparations 147 and 148] were obtained according to a similar manner to that of Preparation 146.
Preparation 147 l-[4-[5-[6--[4-(4-Methyl cyclohexyl)-1-piperazinyll-3pyridyl]-1,3,4-thiadiazol-2-yl]benzoyloxy]-lH-1,2,3benzotriazole IR (KBr) 2943, 2918, 1782, 1601, 1427, 1402, 987, 845 cm'1 NMR (CDCd 3 5) 0. 94 d, J=6. 9Hz) 1. 4-1. 8 (9H, in), 2. 3-2. 5 (lHr in), 2.5-3. 9 (4H, mn), 3. 7-3. 9 (4H, mn), 6. 74 (1H, d, J=9.QHz), 7.4-7.7 in), 8.1-8.5 (6H, mn), 8-74 (1H, d, J=2.311z) MASS (in/z) (Positive): 581.3 Preparation 140 1-4- (1-Cyc-lohe-xyl-1-piperidyl) phenyll imidazo [2,1bj [1,3,4]thiadiazol-6-yllbenzoyloxy-1H-1,2,3-benzotriazole IR (KBr) 2926, 1776, 1608, 1471, 1230, 1176, 980, 845 cm- 1 NMR (ODd 3 8) 1. 0-2. 7 (18H, in), 3. 1-3. 3 (2H, in), 7. 2-8. 4 (13H, m) ()APCT MASS 604.13 Preparation 149 To a solution of 1,4-dioxaspiro[4.5]decan-8-ol (9.5 g) in N,N-dimethylformamide (200 ml) was added portionwise sodium hydride (aft. 60% oil suspension) (2.6 g) under ice-cooling and nitrogen atmosphere. After stirring for 2 hours at room temperature, the reaction mixture was stirred for 1 hour at 6000.
To the reaction mixture was added a solution of 7-iethoxyheptyl 4 -met hylbenzenesulf onate (15.0 g) in N,N-dimethylfornamide ml) at 60 0 C and then the -reaction mixture was stirred for 2 hours at 600C. After being cooled to room temperature, the reaction mixture was poured into a mixture of ethyl acetate and water.
The organic layer was successively washed with water and brine WO 01/60846 PCT/JP01/01204 88 and dried over magnesium sulfate. The solvent was evaporated in vacuo and the residue was purified by column chromatography on silica gel eluting with a mixture of hexane and ethyl acetate The eluted fractions containing the desired product were collected and evaporated in vacuo to give 8-(7methoxyheptyloxy)-1,4-dioxaspiro[4.5]decane (9.77 g).
NMR (CDC1 3 1.3-1.9 (18H, 3.33 (3H, 3.3-3.5 3.94 (4H, s) ESI MASS (m/z)(Positive): 309.3 (M++Na) The following compounds [Preparations 150 and 151] were obtained according to a similar manner to that of Preparation 149.
Precaration 150 8-(8-Methoxyoctyloxy)-1,4-dioxaspiro[4.5]decane NMR (CDC1 3 1.2-1.9 (20H, 3.33 (3H, 3.3-3.5 3.94 (4H, s) ESI MASS (m/z)(Positive): 323.3 (M++Na) Preparation 151 tert-Butyl 4-(6-methoxyhexyloxy)-1-piperidinecarboxylate NMR (CDC13, 1.3-1.7 (12H, 1.45 (9H, 1.7-1.9 (1H, 3.0-3.2 (2H, 3.33 (3H, 3.3-3.5 (4H, m), 3.7-3.9 (2H, m) APCI MASS (m/z)(Positive): 216.07 (M++l-Boc) Preparation 152 A mixture of ethyl 4-[4-4-(7-methoxyheptyloxy)cyclohexyl-1-piperazinyl]benzoate (2.8 g) and hydrazine monohydrate (26 ml) in ethanol (56 ml) and tetrahydrofuran (22 ml) was stirred for 15 hours at 10 0 After being cooled to room temperature, the reaction mixture was poured into water.
The solvent was evaporated in vacuo and the residue was extracted with ethyl acetate. The extract was washed with water and dried WO 01/60846 PCT/JP01/01204 89 over magnesium sulfate. The solvent was evaporated in vacuo to give 4-[4-4-(7-methoxyheptyloxy)cyclohexyl-1piperazinyl]benzohydrazide (2.58 g).
IR (Neat): 2933, 2858, 1608, 1512, 1454, 1240, 1113 cm 1 NMR (CDCl 3 1.2-2.2 (18H, 2.2-2.4 (1H, m 2.62 (4H, 3.33 (3H, 3.2-3.5 (9H, 4.06 (2H, br s), 6.8-6.9 (2H, 7.30 (1H, 7.6-7.7 (2H,m) APCI MASS 447.47 The following compounds [Preparations 153 to 164] were obtained according to a similar manner to that of Preparation 152.
Preparation 153 4-[4-(4-Phenylcyclohexyl)-1-piperazinyl]benzohydrazide APCI MASS (m/z)(Positive): 379.4 Preparation 154 4-[4-(4-Methylenecyclohexyl)-1piperazinyl]benzohydrazine IR (KBr): 3429, 3402, 3307, 3280, 2933, 2837,1608, 1504 cm- 1 NMR (DMSO-d 6 1.20-1.50 (2H, 1.60-2.20 (4H, m), 2.20-2.30 (2H, 2.50-2.70 (4H, 3.10-3.30 (4H, 4.36 (2H, br 4.61 (1H, 6.91 (2H, d, J=8.9Hz), 7.69 (2H, d, J=8.8Hz), 9.45 (1H, br s) MASS 317 Preparation 155 4-[4-(Cyclohexylmethyl)-l-piperazinyl]benzohydrazide NMR (DMSO-d 6 0.70-1.00 (2H, m) 1.10-1.80 (9H, m) 2.11 (2H, d, J=7.2Hz), 2.41-2.46 (4H, 3.19-3.24 (4H, 4.36 (2H, 6.92 (2H, d, J=8.9Hz), 7.69 (2H, d, J=8.9Hz), 9.46 (1H, s) APCI MASS (m/z)(Positive): 317 (M +1) WO 01/60846 PCT/JPO/01204 Preparation 156 4-[4-[4-(8-Methoxyoctyloxy)cyclohexyl]-lpiperazinyllbenzohydrazide IR (Neat): 2931, 2856, 1703, 1608, 1512, 1454, 1240, 1113 cmJ 1 NMR (CDC1 3 1.2-2.2 (20H, 2.2-2.4 (1H, 2.6-2.8 (4H, 3.1-3.5 (9H, 3.33 (3H, 4.05 (2H, br 6.8-6.9 (2H, 7.27 (1H, 7.6-7.7 (2H, m) MASS 461.53 Preparation 157 4-[4-(5-Methoxypentyloxy)-l-piperidylbenzohydrazide NMR (CDC1 3 1.30-1.80 (OH, 1.85-2.05 (2H, i), 3.00-3.18 3.33 (3H, 3.34-3.60 (51, n), 3.60-3.70 (2H, 3.95-4.15 6.88 (2H, d, J=8.95Hz), 7.53 (1H, 7.65 (2H, d, J=8.9lHz) ESI MASS (i/z)(Positive): 358.4 (M++Na) Preparation 158 4-[4-(6-Methoxyhexyloxy)-l-piperidyl]benzohydrazide NMR (CDC1 3 6) 1.3-2.1 (12H, 3.0-3.2 (2H, 3.33 (3H, 3.3-3.5 (5H, 3.5-3.8 (2H, 4.07 (2H, br s), 6.8-6.9 (2H, 7.36 (1H, br 7.6-7.7 (2H, m) APCI MASS (i/z)(Positive): 350.07 Preparation 159 4-[4-(4-Methoxybutoxyethyl)-l-piperidyllbenzohydrazide NMR (CDC1 3 1.2-1.9 (9H, 2.7-2.9 (2H, 3.2-3.5 (6H, 3.33 (3H, 3.7-3.9 (2H, 4.06 (2H, br 6.8-7.0 (2H, 7.30 (1H, br 7.6-7.7 (2H, m) APCI MASS (i/z)(Positive): 335.93 Preparation 160 4-[4-(5-Methoxypentyloxynethyl)-l-piperidyl]benzohydrazide WO 01/60846 PCT/JPO/01204 91 NMR (CDC1 3 1.2-2.0 (11H, 2.7-2.9 (2H, 3.2-3.5 (6A, 3.33 (3H, 3.8-4.2 (4H, 6.38 (2H, d, J-=9.QHz), 7.34 (1H, br 7.6-7.7 (211, m) APCI MASS (m/z)(Positive): 349.93 (M+I) Preparation 161 6-[4-(4-Methylcyclohexyl)-l-piperazinyl]nicotinohydrazide NMR (CDCJ 3 0.94 (3H, d, J=6.9Hz), 1.4-2.0 (9H, m), 2.1-2.3 (1H, 2.5-2.7 (4H, 3.6-3.8 (4H, m), 3.9-4.3 (2H, 6.61 (1H, d, J=9.OHz), 7.43 (1H, br 7.86 (1H, d, J=9.0 and 2.5Hz), 8.54 (11, d, J=2.3Hz) APCI MASS (Positive): 318.00 (Ms+1) Preparation 162 6-[4-(Trans-4-methylcyclohexyl)-l-piperazinyl nicotinohydrazide NMR (CDC1 3 0.88 (3H, d, J=6.4Hz), 0.9-2.0 (9H, rn), 2.2-2.4 (11, 2.6-2.8 (4H, 3.5-3.7 (41, i), 3.9-4.3 (2H, 6.61 (1H, d, J=9.lHz), 7.33 (1H, br 7.85 (1H, d, J=9.0 and 2.5Hz), 8.53 (1H, d, J=2.3Hz) AECI MASS (i/z)(Positive): 318.00 Preparation 163 6-[4-(4-Ethylcyclohexyl)-l-piperazinyl]nicotinohydrazide NMR (CDC1 3 0.8-1.0 (51, 1.2-2.0 (9H, 2.2-2.4 (11, 2.5-2.7 (4H, 3.5-3.7 (4H, 3.8-4.2 (2H, 6.61 (1H, d, J=9.OHz), 7.38 (1H, br 7.86 (1H, dd, J=9.0 and 2.5Hz), 8.54 (1H, d, J=2.4Hz) APCI MASS (Positive) 332.00 (M-l1) PrepQaration 164 WO 01/60846 PCT/JP01/01204 92 6-[4-(Trans-4-ethylcyclohexyl)-1-piperazinyl]nicotinohydrazide NMR (CDC1 3 0.8-2.0 (14H, 2.2-2.4 (1H, 2.5-2.7 (4H, 3.5-3.7 (4H, 3.8-4.2 (2H, 6.61 (1H, d, J=9.0Hz), 7.32 (1H, br 7.86 (1H, dd, J=9.0 and 8.53 (1H, d, J=2.4Hz) APCI MASS (m/z)(Positive): 331.93 (M +1) Preparation 165 A mixture of methyl 4-[2-[4-[4-4-(7-methoxyheptyloxy)cyclohexyl-l-piperazinyl]benzoyl]hydrazinocarbonyl]benzoate (1.9 g) and phosphorus pentasulfide (1.1 g) in ethylene glycol dimethyl ether (40 ml) was refluxed for 1.5 hours. After being added triethylamine, the reaction mixture was successively refluxed for 1.5 hours. After being cooled to room temperature, the reaction mixture was poured into ice-water. Then the solution was adjusted to pH 8 with 1N aqueous sodium hydroxide.
The resulting precipitate was collected by filtration and washed with water to give methyl methoxyheptyloxy)cyclohexyl-l-piperazinyl]phenyl]-1,3,4thiadiazol-2-yl]benzoate (2.13 g).
IR (KBr) 2931, 2856, 1718, 1606, 1439, 1281, 1111, 953 cm-1 NMR (DMSO-d 6 1.0-2.2 (19H, 2.8-3.6 (13H, 3.21 (3H, 3.90 (3H, 7.0-8.3 (8H, m) ESI MASS (m/z)(Positive): 607.4 The following compounds [Preparations 166 to 168] were obtained according to a similar manner to that of Preparation 165.
Preparation 166 Methyl 4-[5-[4-[4-(4-methylenecyclohexyl)-lpiperazinyl]phenyl]-1,3,4-thiadiazol-2-yl]benzoate IR (KBr): 3423, 2939, 2829, 1719, 1603, 1439 cm 1 NMR (DMSO-d 6 1.20-2.00 (6H, 2.10-2.40 (2H, m), WO 01/60846 PCT/JP01/01204 93 2.50-2.70 (4H, 3.15-3.30 (4H, 3.90 (7H, s), 4.62 (1H, br 7.08 (2H, d, J=8.4Hz), 7.85 (2H, d, J=9.3Hz), 7.80-8.30 (4H, m) MASS 477 476 475 (M Preparation 167 4-[5-[4-[4-4-(8-Methoxyoctyloxy)cyclohexyl-1piperazinyl]phenyl]-1,3,4-thiadiazol-2-yl]benzoate IR (KBr) 2929, 2854, 1724, 1606, 1439, 1281, 1111, 955 cm-1 NMR (DMSO-d 6 1.1-2.2 (21H,m), 2.8-3.6 (13H, 3.20 (3H, 3.8-3.9 (3H, 7.0-8.3 (8H, m) ESI MASS (m/z)(Positive): 621.5 Preparation 168 Methyl 4-[5-[4-[4-(5-methoxypentyloxy)-1piperidyl]phenyl]-l,3,4-thiadiazol-2-yl]benzoate NMR (DMSO-d 6 1.00-1.60 (8H, 1.80-1.95 (2H, m), 2.75-2.95 (2H, 3.21 (3H, 3.55-3.75 (7H, m), 3.90 (3H, 7.09 (2H, d, J=8.80Hz), 7.84 (2H, d, J=8.78Hz), 8.13 (4H, s) ESI MASS (m/z)(Positive): 518.2 (M++Na) Preparation 169 A mixture of 4-(l-cyclohexyl-4-piperidyl)benzonitrile (0.68 thiosemicarbazide (0.58g) and trifluoroacetic acid ml) in toluene (7 ml) was stirred for 7 hours at 70°C. After being cooled to room temperature, the solvent was evaporated in vacuo. Then the residue was dissolved in tetrahydrofuran and poured into water. The solution was adjusted to pH 8 with 1N aqueous sodium hydroxide. The resulting precipitate was collected by filtration and washed with water and isopropyl ether to give 5-[4-(l-cyclohexyl-4-piperidyl)phenyl]-1,3,4thiadiazol-2-amine trifluroacetate (0.80 g).
IR (KBr): 3296, 2926, 1632, 1514, 1462 cm 1 NMR (DMSO-d 6 1.0-1.9 (14H, 2.2-2.6 (4H, 2.8-3.0 WO 01/60846 PCT/JP01/01204 94 (2H, 7.2-7.4 (4H, 7.66 (2H, d, J=8.2Hz) APCI MASS 343.20 (M 4 +1) The following compound was obtained according to a similar manner to that of Preparation 169.
Preparation 170 5-[4-[Cis-4-(4-methylcyclohexyl)piperazin-1-yl]phenyl]- [1,3,4]thiadiazol-2-ylamine NMR (CDC1 3 +CDO3D 0.95 (3H, d, J=7.01Hz), 1.45-1.70 (8H, 1.70-1.85 (1H, 2.15-2.30 (1H, 2.65-2.80 (4H, 3.25-3.35 (4H, 6.92 (2H, d, J=8.94Hz), 7.64 (2H, d, J=8.85Hz) ESI MASS (Positive): 358.4 Preparation 171 A mixture of 5-[4-(l-cyclohexyl-4-piperidyl)phenyl]- 1,3,4-thiadiazol-2-amine trifluoroacetate (0.78 g) and ethyl 4-(bromoacetyl)benzoate (0.6 g) in ethanol (15 ml) was stirred for 5 hours at 80'C. After being cooled to room temperature, the reaction mixture was poured into isopropyl ether. The resulting precipitate was collected by filtration, washed with isopropyl ether and added to a solution of trifluoroacetic acid ml) in xylene (15 ml). Then a mixture was stirred for 3 hours at 130 0 C. After being cooled to room temperature, the reaction mixture was poured into isopropyl ether. The resulting precipitate was collected by filtration and washed with isopropyl ether to give ethyl 4-[2-[4-(l-cyclohexyl-4piperidyl)phenyl]imidazo[2,1-b][1,3,4]thiadiazol-6yl]benzoate trifluroacetate (0.45 g).
IR (KBr): 2941, 1701, 1676, 1610, 1471, 1279, 1200, 1180, 1132 cm- 1 NMR (DMSO-d 6 1.0-2.2 (17H, 2.8-3.4 (4H, 3.4-3.6 (2H, 3.8-4.5 (2H, 7.2-8.1 (8H, 8.94 (1H, 9.16 (1H, br s) WO 01/60846 PCT/JPOI/01204 ESI MASS (Positive): 515.3 The following compound was obtained according to a similar manner to that of Preparation 171.
Preparation 172 Ethyl 4-[2-[4-[cis--4-(4-methylcyclohexyl)-1piperazinyllphenyl]imidazo[2,1-b][1i,3,4thiadiazol-6yl]benzoate bis (trifluoroacetate) NMR (CDCl 3
+CD
3 OD, 8) 1. 00 (3H, d, J=7.l13Hz) 1. 42 (3H, t, J=7.llHz), 1.50-2.10 (10H, mn), 2.90-3.20 (4H, mn), 3. 55-3. 80 mn), 4. 39 (2H, q, J=7. 22Hz) 6. 97 (2H, d, J=8.89Hz), 7.78 (2H, d, J=8.78Hz), 8.10 (2H, d, J=8.42Hz), 8.11 (1H, s) ESI MASS (Positive): 529.7 Preparation 173 To a solution of benzyl 4-(trans-4cyclohexylcyclohexyl) -1-piperazinecarboxylate (4 g) in ethanol (40 ml) and dioxane (40 ml) was added 10% palladium on carbon (0.8 and hydrogen gas at atmosphere pressure for 7 hours.
To the reaction mixture was added dichloroinethane (40 ml) The reaction mixture was filtered, and the filtrate was concentrated by evaporation under reduced pressure to give l-(trans-4cyclohexylcyclohexyil)piperazine (1.56 g).
IR (KBr): 1446, 1140, 835 crrJ 1 NMR (CDCl 3 6) 0.75-1. 35 (l2H, mn), 1. 5-2. 25 (9H, in), 2. 54 (4H, t, J=4.8Hz), 2.B9 (4H, t, J=4.BHz) MASS 251 The following compound was obtained according to a similar manner En that of Preparation 173.
Preparation 174 1- (Trans-4-tert-butyicyciohexyl) piperazine WO 01/60846 PCT/JPOI/01204 96 IR (KBr): 1450, 1365, 1140 cm'1 NMR (CDC1 3 6) 0.-8 4 (9H, s) 0. 8-1. 35 m) 1.7 25 in), 2.54 (6H, t, J=4.8Hz), 2.89 (4H, t, JTh4.9Hz) MASS 225 Preparation 175 To a mixture of cesium carbonate (2.54 palladium(TI) acetate (62 mng) and 2,2'-bis(diphenylphosphino)-1,1'binaphthyl (260 mg) in toluene (6 ml) was successively added methyl 4'-trifluoromethylsulfonyloxy-1, 1'-biphenyl-4carboxylate (1 g) and 1-(trans-4-cyclohexylcyclohexyl)piperazine (835 mg) in stream of nitrogen. The mixture was stirred at ambient temperature for 45 minutes and at 110 0 C for further 23 hours. After cooling to room temperature, water and dichioromethane was added to the reaction mixture. The resulting precipitate was collected by filtration and washed with water and dried to give methyl 4'-[4-(trans-4cyclohexylcyclohexyl) -1-piperazinyl] -biphenyl-4carboxylate (604.0 g).
NMR (CDCl 3 0.8-2.4 (21H, mn), 2.65-2-8 (4H, in), 3.2- 3. 4 (4H, in), 3. 93 (3H, s) 6. 99 (2H, d, J=8.8Hz) 7. (2H, d, J=8.9Hz), 7.62 (2H, d, J=8.4Hz), 8.06 (2H, d, J=8. 3Hz) MASS 461 The following compounds [Preparations 176 to 179] were obtained according to a similar manner to that of Preparation 175.
Preparation 176 Methyl 4'-[4-(Trans-4-Lert-butylcyclohexyl)-lpiperazinyll -biphenyl-4-carboxylate NMR (CDCl 3 0.86 (9h, 0.9-1.4 1.7b-2.4 in), 2.75 (4H, t, J=4.9Hz), 3.28 (4H, t, J=4.9Hz), 3.93 (3H, 7.00 (2H, d, J=8.8Hz), 7.56 (2H, d, J=8.7Hz-), WO 01/60846 PCT/JPOI/01204 97 7.62 (2H, d, J=8.4Hz), 8.06 d, J=8.3Hz) MASS :435 Prep)aration 177 tert-Butyl 4-(4'-methoxycarbonyl-1,1'-biphenyl-4-y)piperazinecarboxylate NMR (CDC1 3 1.49 (9H, 3.15-3.25 (4H1, mn), 3.55-3.65 (411, in), 3.93 (31H, 6.99 (2H1, d, J=6.8Hz), 7.65 (4H, mn), 8.06 (211, d, J=6.8Hz) MASS 396 (M++23) Prelparation 178 Methyl 4'-[4-(4-methoxyphenyl)-l-piperazinyl]-1,1'biphenyl-4-carboxylate NMR (CDC1 3 3.2-3.35 (4H, mn), 3.4-3.5 (4H, mn), 3.79 (3H, 3.93 (3H, 6.8-7.1 in), 7.5-7.7 (411, in), 8.07 (2H, 6, JTh8.3Hz) MASS 403 Preparation 179 Methyl 4'-[4-(4,4--dimethylcyclohexyl)-1--piperazinyl]- 1,1' -biphenyl-4-carboxylate NMR (ODC1 3 0. 92 (6H1, s) 1. 1-1. 85 (811, in), 2.1-2. 3 (111, 2.7-2.85 (4H1, in), 3.2-3.4 (4H, mn), 3.93 (3H1, s), 7.00 d, J=8. 8Hz) 7. 56 (2H, d, J= 8.8Hz) 7. 62 (2H, d, J=8.411z), 8.06 (2H, d, J=8.4H2l) MASS (in/z) 407 Preparation 180 To a solution of 2- (hydroxyinethyl) -1,3-propanediol 0 g) and dimethoxymethyl) ben zoec (6.36 ml) in DMF (50 ml) was added acid (1.31 g) and the mixture was stirred at amnbient temperature overnigjht. To a -reaction mixture were added triethylainine 18 ml) and water (150 ml) and the solution was extracted twice with ethyl acetate (150 ml). The extracts WO 01/60846 PCT/JP01/01204 98 were washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture of hexane and ethyl acetate (1:1 The fractions containing the object compound were collected and evaporated under reduced pressure to give (2-phenyl-1,3-dioxan-5-yl)methanol (4.51 g).
NMR (CDC1 3 3.43, 3.46 (1H, each 3.68-3.79 (2H, m), 4.00, 4.04 (1H, each 4.10-4.27 (3H, 5.41, 5.51 (1H, each 7.30-7.50 (5H, m) Preparation 181 To a solution of (2-phenyl-l,3-dioxan-5-yl)methanol g) in dichloromethane (40 ml) were added pyridinium chlorochromate (11.6 g) and molecular seaves 4A powder (5.0 g) with stirring and the mixture was stirred at ambient temperature for 5 hours. To a reaction mixture was added dichloromethane (100 ml) and the insoluble material was filtered off with celite and the filtrates were washed in turn with water, saturated aqueous sodium hydrogen carbonate and saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo to give 2-phenyl-l,3-dioxane-5-carbaldehyde (0.52 This compound was immediately used as the starting compound for the next step.
Preparation 182 Anhydrous cerium(III) chloride (10.0 g) was added to THF (100 ml) with stirring under ice-cooling and a mixture was stirred at ambient temperature overnight and then cooled in an ice bath. A solution of cyclohexyl magnesium chloride (2M solution in diethyl ether) (20.3 ml) was dropwise added to the mixture with stirring on ice bath (keeping the temperature below To the mixture was added dropwise a solution of tertbutyl 4-oxo-l-piperidinecarboxylate (5.38 g) in THF (25 ml) and the mixture was stirred at 0-6°C for 1 hour. 10% aqueous acetic acid (100 ml) was added to the reaction mixture and extracted WO 01/60846 PCT/JP01/01204 99 twice with ethyl acetate (100 ml). The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (400 ml) eluting with a mixture of n-hexane and ethyl acetate (2:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give tert-butyl 4-cyclohexyl-4hydroxy-1-piperidinecarboxylate (7.27 g).
NMR (CDC1 3 0.90-1.30 (5H, 1.46 (10H, 1.49-1.90 (9H, 2.90-3.15 (2H, 3.50-3.70 (1H, m), 3.80-4.00 (2H, m) ESI MASS (m/z)(Positive): 306.3 (M++Na) Preoaration 183 To a solution of tert-butyl 4-cyclohexyl-4-hydroxy-lpiperidinecarboxylate (7.26 g) in DMF (70 ml) was added sodium hydride (60% in oil) (2.05 g) with stirring under ice-cooling.
The mixture was stirred at ambient temperature for 1 hour. To the suspension was added methyl iodide (4.79 ml) and the mixture was stirred at ambient temperature overnight. The reaction mixture was poured into ice-water (300 ml) and extracted three times with ethyl acetate (200 ml). The extracts were collected, washed twice with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (400 ml) eluting with a mixture of n-hexane and ethyl acetate (4:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give tert-butyl 4-cyclohexyl-4methoxy-1-piperidinecarboxylate (6.77 g).
NMR (CDC1 3 0.85-1.30 (5H, 1.45 (10H, 1.46-1.85 (9H, 2.85-3.10 (2H, 3.12 (3H, 3.80-3.95 (2H, m) ESI MASS (m/z)(Positive): 320.3 (M-+Na) Precaration 184 WO 01/60846 PCT/JP01/01204 100 To a solution of tert-butyl 4-cyclohexyl-4-methoxy-lpiperidinecarboxylate (2.04 g) in a mixture of dichloromethane ml) and anisole (5.2 ml) was dropwise added trifluoroacetic acid (10.6 ml) with stirring under ice-cooling. The mixture was stirred at ambient temperature for 1 hour and then concentrated in vacuo. The resulting residue was azeotropically distilled three times with toluene (20 ml) and dried in vacuo. The obtained residue was dissolved in DMSO (20 ml) To the solution were added ethyl 4-fluorobenzoate (2.60 g) and potassium carbonate (2.84 g) and the mixture was stirred at 140"C overnight. The reaction mixture was poured into water (100 ml) and extracted twice with ethyl acetate (80 ml) The extracts were collected, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated in vacuo. The resulting residue was chromatographed on silica gel (200 ml) eluting with a mixture of n-hexane and ethyl acetate (9:1 The fractions containing the desired compound were collected and evaporated under reduced pressure to give ethyl 4-(4-cyclohexyl-4methoxy-1-piperidyl)benzoate (1.81 g).
NMR (CDC13, 0.85-1.32 (5H, 1.36 (3H, t, J=6.80Hz), 1.50-1.90 (10H, 2.95-3.15 (2H, 3.16 (3H, s), 3.55-3.70 (2H, 4.32 (2H, q, J=7.12Hz), 6.86 (2H, d, J=9.12Hz), 7.90 (2H, d, J=9.08Hz) ESI MASS (m/z)(Positive): 368.3 (M++Na) The following compound was obtained according to a similar manner to that of Preparation 152.
Preparation 185 4-(4-Cyclohexyl-4-methoxy-l-piperidyl)benzohydrazide NNR (CDC1 3 0.9-1.9 (15H, 3.0-3.2 (2H, 3.16 (3H, 3.5-3.7 (2H, 4.06 (2H, br 6.8-7.0 (2H, m), 7.33 (1H, br 7.6-7.7 (2H, m) APCI MASS (m/z)(Positive): 332.40 WO 01/60846 PCT/JPOI/01204 101 The following compound was obtained according to a similar manner to that of Preparation 32.
Preparation 186 M'~ethyl 4- (4-cyclohexyl-4-methoxy-lpiperidyl) benzoyl] hydrazinocarbonyll benzoate NMR (CDCl 3 6) 0. 9-2. 0 (15K, in), 3. 0-3.2 (2H, in), 3.17 (3H, 3.5-3.8 (2K, in), 3.95 (3H, 6.8-7.0 (2K, mn), 7.6-7.8 (2H, mn), 7.8-8.0 (2H, in), 8.0-8.2 (2H, mn), 9.1-9.2 (1H, mn), 9.5-9.7 (1H, m) ()APCI MASS (Positive) 494.47 The following compound was obtained according to a similar manner to that of Preparation 47.
Preparation 107 Methyl 4-[5-[4-(4-cyclohexyl-4-inethoxy-lpiperidyl) phenyll 4-thiadiazol-2-yl] benz-oate ESI MASS (m/z (Positive): 492.3 (M++Il) The following compound was obtained according to a similar manner to that of Pre-paration 82.
Preparation 188 4-[5-[4--(4-Cyclohexyl-4-rnethoxy-l-piperidyl)phenyll- 1,3, 4-thiadiazol-2-yllbenzoic acid NMR (DMSO-d 6 6) 8 (15K, in), 2. 9-3.2 (5K, in), 3. 6-3.8 (2H, in), 6.9-7.2 (2H, mn), 7.7-8.3 (6K, m) EST MASS (in/z) (Negative): 476.1 (M-l1) The following compound was obtained according to a similar manner to that of Preparation 112.
Prep2aration 189 (4-Cyclohexyl-4-methoxy-l-piperidyl)phenyl1- WO 01/60846 PCT/JP01/01204 102 1,3,4-thiadiazol-2-yl]benzoyloxy-1H-1,2,3-benzotriazole IR (KBr): 2927, 1784, 1603, 1441, 1412, 1234, 1192, 1080, 987 cm-1 NMR (CDC1 3 0.9-1.9 (15H, m) 3.0-3.3 (5H, m) 3.5-3.8 (2H, 6.9-7.1 (2H, 7.4-7.7 (3H, 7.90 (2H, d, J=8.9Hz), 8.1-8.3 (3H, 8.3-8.5 (2H, m) ESI MASS (m/z)(Negative): 476.1 (M--HOBT-1) The Starting Compounds used and the Object Compounds obtained in the following Examples 1 to 95 are given in the table as below, in which the formulas of the starting compounds are in the upper column, and the formulas of the object compounds are in the lower column, respectively.
to be continued on the next page WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HO ()IF bo HOH HO' IH 0 CH3 O H E o -a3O
'OH
N aeS
HO
major HO 0 OH minor
G
HO13,.6 H..HC.H 0
OHB
3 N f 0 H O o
HN)-
HO' NH oc HO OH J C3 HO' H H 3H 0 M &OH 0 OHH Na 03S0 HO 0
.P
HO H3C,. Nhlq NHBoc 0bHO' H HOH3 0 0 H 2
O
OH
HOH
0 'O %J&OH HO~O 0 0 WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HO B-1 0 HO NC) NH 2 0 0O H T O) OH 3 0 OH oH HO HO 0 ,P HO 3 N O(CH 2 5OCH 3 0 HO H 0 7"H3 0H HO3SO- HO HO 0 OH 0
N
HO J_ NZ (CHi2) 0 H3
OO
'DH
H03SO
O/
HO OH o
HOC
HO HD 0 0 HP'H3 0
H
OHH
H3SO
HO
WO 01/60846 PCT/JPOI/01204 105 Example No. Formula
HOH
H 3 OH~o NC-\ OH HO, 1H 0 H
OHH
NaO3SO
H
OH
HO 0 f o H3C,> NHBoc -NH
HIOH
HO H ~7H3 ~OH ~QOH NaO 3 SO
HO
HO 0 9 0 NHBoc oO H O=?JOH3
~OH
NaO3SO
OH
HO
ninor Maj or WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HO
H
H03S0 O
HO
V
OH 0 HO 0 HN o~ 1~H[ HH oH
H
3 N HO H 0 "b>O H03SO /R INK QCH 3
HO
OH 0 HO 0
H
\N
OH
HO01 0 O N' OH
Q
H03SO OR /CH3
HO
WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula OH] 0 H3C,&lqlq o OH H2N O ~0 H3 N O 'OH
HD
9 OH 0 HO 0 H$O
NN
H H3C-
H
HO O H 'r
H
3 SO OH
HO
OH 0 HO 0 ~~K~)(CH)CH3
H
3 C-JN711N H2N 0 H 0 OH HH 0 CH 3
*OHH
H03SO
OH
.HO
OH 0 HOO. K C Ji-(CH2) 1 4 CE3 HO H *3C 0 q~H HO 0 0 >O ODH X0 HO3SO
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula 0- 0 1,(H2 4CH3- CH2) 2 4 CH3 OH 0 14OH 3 WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. I Formula
HO
HD
H03
I
(CH2) 70CH3 H03SI WO 01/60846 WO 0160846PCT/JPO1!0120S Formul a CI]2) 70CH3 HO3SO.
(CH-2) 70CE3 (CH2) 6 0CE 3 WO 01/60846 PCT/JPO1!01204 Formula /7N~s6 NaO3: 70CH3 WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
HO HO0 H H3S 3 ,OHH)71 H pOH 0 HO HO0 H-
HO
3 SON HN< OH H0 S OH C) H 3 N CH2) 40 CH3 HO: o HPO H
B-O
3 HO H 0 C H 3 ]!MOCO P
-OH
0 OH H03SO
HO
0 OHo H3C,. e\Nd-V\
O(CH
2 40CH3 HO a H19 HO- H HH 0= 2 c H 3
~OH
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula OH 0 HO 0 N1 N' (CH 2 6OMe HO o HP On H I HO" O=0 H3
O
21 HO 0 0 H 3C, N4Cj N /f T'4-C-S (CH2) H 0 HPO
OH
HO3SO Ho OHl 0 HO H 3 C, Hno 0 /c NL& -c HO H 000
OH
HOH
Na03S0
~OH
H HOS 0 f/ H HOC WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula HO 0
'-P
H C2 60CH3 HO j HO< 0 0A&QO(HH 0H' H O7CH 3 l- O H PO NaO3SO
HO
23
OH
HO 0 HO N'r'H '-IF HO: H c HNOH H0 H O= 7'CH 3
OH
HO3SO k/
HQ
HO 0 O H 0 s-i HO H HO' H 0 CH3 W-0o
'.OH
H
H03SO
HO
24 HO C~ 2 0 rt 9H i-
HOH
HOR o HP O H H
H
N 'O
OHH
H0330
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No.
Formula
I
N-0 H
O(CH
2 6 0CH 3
H
OH
'CH j -Nr (CH2) 6 0CH 3 (CH2) 26 N-0 -l WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula HO 0
OH
N1 0 J OH 0 'OH RN
O
OHH
HO 0 eH
Q
HO OHNI OH
HO
JfH 0 -H3
OHH
HO3SO-
HO
HO PH0 HO 0 oH OH H03i HO" 0 H3 0[4 0 OH bH H03S0 28 HO 0 iO HO H3 -d0 0G
OHL=O
H~ '0 7CH3 0 7
OH
H0330\\/
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Formula HO 0
.P
,_&Ng-\N'NI HOC 0
P
oHbf OH (CH2 400113 CH2) 40CH3 H-0 3 S0.
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
H-1 0
F
HO H Nd7\ NH2 Na HI OH H0 0H 4 H 0 CR3,(
OH
HO
3 SO ~e
OC
31 HO 0 HO'X H 0 R 0 HXO N "OH a (H603 NaO3SO/
THO
LN CH2) 4 0CH3 WO 01/60846 PCT/JPOI/01204 119 Example No. Formula
OH
HO~ 0f
H
3 C. /NH2 HO oHI OH HO) IfH O 0H H 1C- OF H03SO
HO
33
OH
HO 0 0NN%/\(H
OH
H
3 N 2 4
H
OHH
HOS
,O
HO'O
HODHO o HO3 oO
HO
HO
OH
HO 0 NH2 -1
.OH
HO OSO 3
H
34 S3 WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HO 0 H3 C-y d NH2 ~HO 0 H DO HC H H 0 CH3 '0
C~H
HO
3 SO
OH
O
P
HO 0 HO 6 H09
HNOH
HH H 0 CH3 H03SO H80.O
H
HO
OE
HO 0 HH3C-,N 0H2 HOJ HO'H f OHk C H3 OH
'OH
H0 3
SO
36
HO
HO 0 OH 0 3% <-Nfi\ N' C H HO o H OH IJ HO H 0 C H3 0?
OH
H(O3SO
OH/
HO
WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HO 0
H
3 c2 6 NH2 HO 0H
O
HO fH O HO' 0 H3
UH
H03SO OH )0 37 HO 0 *OH o HO0HO
OO
HO
HO, HO" 0~ H3 O Ib)H OH H03SO
OH
HO C HO 0. HI 0OH HO..H."HO= H3
O'
H03SO 38 OHo HO 0 s HO H3C oH 0 Ho HO, H 0 CH3 -4 OH X-
OH
H03SOK 0\-)Z WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formaul a HOJV~ 0 HO H r N/H? -o OHP1)OH OHH 3 1-'OH 0 1 HO3S H OH -O D H HH 0
HO
"_H3C, 0N
H
HOH
HO
0 HH0O=OCH3
OHH
HO3SO- D D 0 400 ~~HO 0OH NNDG~(H2 )OH H H$OH o 0O 0 \-OH j H0 H3 -1O 01- 400 WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
HO 0 NH2 HO H 0o O H3 Fmoc
"'OH
OH
H03SO
H
41 H~C.2'-J N \N N(~jNC:--O (CH 2 4OCH 3 HO oHc..
YH
H9.HO" 0 H3 0 Fmoc N N)H H035_
ID/
HO
HO 0
OH
HO 3-d 0 1) -O H OH WO 01/60846 124 Example No. Formula
OH
HO 0 H3C_6NIV
HO
43 PCT/JPO1!0120S WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formnula- NE~oc H2N o B OH N
"OH
D H H03SO
H
HO G 9 HO-N o HN1 0OH
OO
HO
3 SO_ _e/ Ho
OH
HO 0 0 N-N H2N N~Oh-skKD (CH 2) -7 OCH3
'OH
HO-H
460
OH
HO
HO46O7'H
OH
OHH
HO
WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH-
HO 0 f0 N-N .lOH
OR
H0 3 SO Ho 47
OH
HO 0 N-N H C. 6N4 -s&G (CH2)7OCH3 cH$OH 0
-OH
H03SO
HO
HO 0 0 N- H~c2N O (CH2) 700H3 H2N 0 HP OH H 0 Ho7'H3
'OH
OR
HO3SO
HO
48
OH
HO 0 0 N-N OH N1 (CH2) 70CH 3 6H OH o H OHl H 0 H 0 A
OHH
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
HO 0 F 0 N-N H2N o§ H OH
OH
HO 0H p- H3O /Y5~4--N-OCH)O O NHO 0\1 H0 H 61 HNO HO o H O 7H3 0 H3
OHH
HO
H OH HO 0 H0 H 3C-y -H I HjJ N OH N-" HO H 0 O CH3 H03SO
HO
HO 0 HO HO HO 7 O(CH2)7CH3 HO- BOHN O H.C OH
H
OHH
O
H
H03SO
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No.
Formula f HO 0
H
3 C
N
H
H0 3
SO
51
HO
HO3.
HC
H0 3 S0 -r\
I
1-1 0 H 3NJ OHo N)1' N C(CH 2) 50 H 3 CH2) 50CH3
I
WO 01/60846 WO 0160846PCT/JPO1!0120S Formula 70CH3 H 3 S 0- CH2) 70CH3 S-0 2N WO 01/60846 PCT/JPOI/01204 130 Example No. Formula
OH
HO 0 N4- NH 2 HO 0 HN OH
HO
3 0 yN'OH 0O 3 SO
HO
OH o HO O(
HOH
HO o HO 0 NH 2 s
H
a NOH HO NHO~ H
HO
3 S 0
HO
H
3 C WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula HO 0 HO d\
H
0 1 HO o H N
OH
HO
Y
H0 3 SO _e 57 HO I OH 0
C
HO 0 ~Q N-N~ HO H HO3 o HN OH
OHH
HD
3 SO HO0
HO
HO
0(CH 2 7 Ome 0 (CH 2 7 OMe
HO
3 Major
HO,
HO.
HO
3 Minor WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
I
(CH
2 8 O~e
HO
HOa Maj or
HO
3
SC
Minor
I
(OH
2 BOMe HOC 0~\N HO tN HNCOH N HO' H o~7
OHOH
HO
HOC
OH
0 a ~NO~NZ-oN
I
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No.
Formula
OH
HO0 HO 0 OH N- DO(H)O0 HO0 HO o HN OH 0 H 3 SO O H"O
HO
CR
2 6 0CH 3 WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
HO 0
H
2
HO
HOa
(OH
2 4 0CH 3 (CH 2 )5 5 CH 3 WO 01/60846 PCT/JPOI/01204 135 Example No. Formula
OH
HO 0 z HO LNd- NH 2 HO o HN OH 0 r <VHO' H0 COO
O
HO OH0 HO 0 N-N HO V- oHN OH 0- HO C H
HO
3
SO_?
HO
OH
HO 0 HO 0HN OH 0~ HO 0 66
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S 136 Example No. Formula HO 0 HO a )H 0 0
HO
3 SO 67
HO
HO d H HO Q HN OH H- HO H O 0
HO
3 SO
H
HO
HO0 HO
YH
HOa a 0
HOH
68
N-N
WO 01/60846 PCT/JPOI/01204 137 Example No. Formula
OH
HO 0 HO (yNdF NH 2 HO 2H O 0 HO HNO
HO
3
SO
69
HO
1-10 0 'OH o HO H N1P HO 'A0HN 7H 11H HO 0
H
N
'OH
-OH
HO
OH
HO NdrC\
NH
2
HO
HO 'Ao HN OH N 1 10H H0 3 S0
HO
HO
HOa WO 01/60846 WO 0160846PCT/JPO1!0120S Example No.
Formula {~O.0
I
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula
OH
HO 0 x HO H 3 0 6 QNJF\ NH 2 HO 'A 3 0H OH
.HO
3 SO
OH/
73
HO
HO 0 OH
N
HO H 3 C, HOa 0 H O H liHO 0
CH
COH
HO
3 SO_
?O
HO
OH
HO 0 HO H 3 ~YLN NH 2 HO oA H N OH Fm C H 0
H
H
"OH
HO
3
SOO
74 RO HO
-H
HO H3 DHI*-Q)\ HO 'A0 HN OHi H HO' H0 CH
-OH
HOH
HO
3 SO
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula I rn HO 0) O~e q N HO H.
HOa
HO
3
SO.
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula OMe
-KD-KD
q N OOe HO Ii WO 01/60846 PCT/JPO1!0120S Example No.
79 Formula HO 0 I I N OMe WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula O~e
N-,
HO
3
SO.
WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula HO 0
HO
HO
OH
HO 0 HO Y "Nr~ NH 2 HN OH
OHN
HOSO
HO
4 N
N
HO1
HO,
WO 01/60846 WO 0160846PCT/JPO1!01204 Example No. Formula
OH
HOO
HO HN OH HOOa OH Oe ?bH
N
HOO
HO 0 HN, OH HO HHO k H OH
OH
H0 3 S0 ~e
HO
HO0
H
3
C-.UNI
HO 0 HO Fmoc' O
NH
2 HO S0- WO 01/60846 WO 0160846PCT/JPO1!0120S Example No. Formula HO HO 00Ome HO~a H 3 C..y -oj 1 N H OHN OH H HU~ Hi 0 CH 3 O JOH H0 3
S(
I
OH
2 14
CH
3 H 2
N
HO
3
SO-
OH
2 14
CH
3 WO 01/60846 PCT/JPO1!01204 Example No. Formula OH 0 HOO/~ I HO> N oH HOH H O1*C-H HO H
NH***OH
OH 6i 0 H03SO--J bHO
HOO
89
HO
OH 0
HO
H
3 C. 0 N 2 1
>CH
2 1 4
CH
3 HO Fmoc HOii N H N OH H 0 H 3~CH
HOO
HN
OH
HO
OH 0 H HO OC,. H -00 N.CH CH 3 HOX NF a HN 0 H H' H
OCH
3 O
H
N
-OH
H0 3 SO O
HO
OH 0 H HO ONd-- NA1CH C HO H HO Boc dNH HO>- N HN 0OH HO H 0 CH 3 0O H
OHO
NaO
HO
WO 01/60846 WO 0160846PCT/JPO1!0120S Formula H 3 s 0-
H
0
N,
:xj
HO
3
SO.
WO 01/60846 PCT/JPO1!0120S 149 Example No. Formula
OH
HO 0 H ~NHBoc; HO>0 HOH c) H CH HO 3 So 93 110 WO 01/60846 PCT/JP01/01204 150 Example No. Formula
OH
HO H O NH NH 2
HO
H 0 3 S O
HO
HO -OH 0 N-N OMe HO Nd\ N-'KQ,-4= HO l O 0) HO3 S0_, HN OH H 0 -T N
OH
HO
3 SO- 0
HO
Example 1 A solution of the starting compound (4.42 g) and palladium on carbon (50% including water) (3.0 g) in a mixture of methanol (90 ml) and water (80 ml) was hydrogenated under an atmospheric pressure of hydrogen with stirring at ambient temperature for 8 hours. To the reaction mixture was added palladium hydroxide on carbon (50% including water) (4.0 and the mixture was hydrogenated under an atmospheric pressure of hydrogen with stirring at ambient temperature for 16 hours. The catalyst was filtered off and washed with a mixture of methanol and water (1:1 v/v) (50 ml), and the filtrate' and washes were combined. To the solution was dropwise added allyloxycarbonyl chloride (1.72 ml) in tetrahydrofuran (4 ml) adjusting to pH 8.5-10.0 with IN sodium hydroxide with stirring on an ice-bath.
The mixture was stirred at the same temperature for 2 hours and WO 01/60846 PCT/JP01/01204 151 adjusted to pH 8.0 with IN hydrochloric acid. The solution was subjected to column chromatography on ODS (Daiso-gel, SP- 120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (400 ml) eluting with 10% acetonitrile in water and then with acetonitrile in water. The first fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the major object compound (0.47 The second fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the minor object compound (2.91 g).
major object compound (1) IR (KBr): 1761, 1672, 1635, 1512, 1450 cm- 1 NMR (DMSO-d 6
D
2 0, 0.96 (3H, d, J=6.79Hz), 1.00-1.15 (3H, 1.35 (9H, 1.45-2.50 (9H, 2.80-3.40 (6H, 3.70-4.60 (16H, 4.65-4.90 (4H, m), 5.10-5.45 (4H, 5.80-6.10 (2H, 6.71 (1H, d, J=8.23Hz), 6.77 (1H, d, J=9.01Hz), 6.98 (1H, s) ESI MASS (m/z)(Positive): 1277.2 (M++Na) minor object compound (1) NMR (DMSO-d 6
D
2 0, 0.96 (3H, d, J=6.57Hz), 1.06 (3H, d, J=4.94Hz), 1.36 (9H, 1.45-2.45 (8H, m), 2.75-3.70 (9H, 3.75-4.60 (12H, 4.69 (2H, d, J=5.19Hz), 4.70-4.90 (2H, 5.05-5.50 (3H, m), 5.80-6.10 (1H, 6.91 (1H, d, J=8.29Hz), 7.10 (1H, d, J=8.31Hz), 7.43 (1H, s) ESI MASS (m/z)(Positive): 1193.3 (M++Na) Example 2 A suspension of the object compound (1.73 g) in dichloromethane (40 ml) was stirred with cooling at 5 0 C and treated with triethylsilane (1.1 ml), followed by trifluoroacetic acid (3.19 ml) dropwise over 30 minutes. After WO 01/60846 PCT/JP01/01204 152 warming to room temperature, the clear solution was stirred for 2 hours, and then poured into a mixture of saturated aqueous sodium hydrogen carbonate (100 ml) and pH 6.86 standard buffer (100 ml). Organic solvent was removed by evaporation, and the remaining aqueous solution purified by ODS (Daiso-gel, SP- 120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting with aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (1.10 g).
IR (KBr): 1761, 1668, 1647, 1539, 1512, 1437 cm- 1 NMR (DMSO-d 6
D
2 0, 0.95 (3H, d, J=6.77Hz), 1.18 (3H, d, J=4.94Hz), 1.40-2.40 (7H, 2.70-3.40 (4H, m), 3.60-4.60 (17H, 4.69 (2H, d, J=5.37Hz), 4.70-4.90 (2H, 5.10-5.50 (4H, 5.80-6.20 (2H, 6.89 (1H, 7.08 (1H, d, J=8.21Hz) ESI MASS (m/z)(Positive): 1155.4 (M++Na) Elemental Analysis Calcd. for C 46
H
68
N
8 0 23 S-4H 2 0: C 45.84, H 6.36, N 9.30 Found C 45.85, H 6.33, N 9.16 Example 3 A solution of the starting compound (0.43 g) in dimethylformamide (4 ml) was treated with methoxypentyloxy)piperidin-1-yl]phenyl]imidazo[2,1thiadiazol-6-yl]benzoic acid benzotirazol-1-yl ester (194 mg) and diisopropylethylamine (78.4 R1) and stirred for 5 hours at room temperature. Ethyl acetate (50 ml) was added, and the resulting precipitate collected, washed with isopropyl ether, and dried to give the object compound (610.6 mg) as a crude powder, that was used directly in the next reaction without purification.
WO 01/60846 PCT/JP01/01204 153 Example 4 To a solution of the starting compound (610.6 mg) in a mixture of methanol (10 ml) and tetrahydrofuran (25 ml) were successively added triphenylphosphine (32 mg), tetrakis(triphenylphosphine)palladium(0) (35 mg) and morpholine (106 1l) with stirring, and the mixture was stirred at ambient temperature for 3.5 hours. Ethyl acetate (100 ml) was added, and the resulting precipitate collected, washed with isopropyl ether, and dried to give a crude pale yellow powder (535 mg). The crude powder was dissolved sodium hydroxide aqueous solution and subjected to column chromatography on ODS (YMC-gel ODS-AM-S-50 (Trademark: prepared by YMC Co., Ltd.)) (37% acetonitrile aqueous solution). The fractions containing the object compound were combined, and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (293.7 mg).
IR (KBr): 3355.5, 1633.4, 1608.3, 1529.3, 1517.7, 1463.7, 1444.4, 1267.0, 1230.4 cm 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.10 (3H, d, J=5.6Hz), 1.2-5.6 (65H, 6.71 (1H, d, J=8.1Hz), 6.78 (1H, d, J=9.7Hz), 7.00 (1H, 7.09 (2H, d, J=9.1Hz), 7.75 (2H, d, J=8.7Hz), 7.95 (4H, 7.3-8.7 (7H, 8.79 (1H, s) MASS 1465.5 Elemental Analysis Calcd. for C 66
H
90
N
12 0 22
S
2 -7H 2 0: C 49.74, H 6.58, N 10.55 Found C 49.72, H 6.43, N 10.40 Example A solution of the starting compound (10 g) in a mixture of methanol (500 ml) and water (100 ml) was treated with cobalt (II) chloride hexahydrate (9.43 g) and then stirred to give a pink solution. Sodium borohydride (7.5 g) was then added portionwise and stirred for 1 hour at ambient temperature. The reaction mixture was filtered through a bed of celite, washing WO 01/60846 PCT/JP01/01204 154 with a mixture of methanol (100 ml) and water (20 ml). The ice-cooled filtrate was then treated dropwise with a solution of allyloxycarbonyl chloride (1.46 ml) in tetrahydrofuran ml) keeping pH 8.0-9.5 with IN sodium hydroxide and then stirred for 1 hour at the same temperature. The reaction mixture was evaporated in vacuo (about 200 ml) and added IN sodium hydroxide ml), and then the mixture was stayed in the refrigerator overnight. To the solution was added water (200 ml), and the mixture was purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting with aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (8.58 g).
IR (KBr): 1670, 1633, 1516, 1443, 1269 cm 1 NMR (DMSO-d 6 D20, 0.97 (3H, d, J=6.75Hz), 1.08 (3H, d, J=5.52Hz), 1.35 (9H, 1.40-2.00 (6H, m), 2.10-2.50 (3H, 2.80-3.40 (4H, 3.65-4.50 (14H, 4.65-4.85 (2H, 5.05-5.35 (2H, 5.70-6.00 (1H, 6.72 (1H, d, J=8.12Hz), 6.78 (1H, d, J=10.1Hz) ESI MASS (m/z)(Positive): 1119.3 (M++Na) Elemental Analysis Calcd. for C 4 5
H
67
N
8
O
2 1 SNa-5H 2 0: C 44.52, H 6.37, N 9.44 Found C 44.59, H 6.43, N 9.47 Example 6 A suspension of the starting compound (8.5 g) in dichloromethane (180 ml) was stirred with cooling at 5°C and treated with triethylsilane (6.2 ml), followed by trifluoroacetic acid (17.9 ml) dropwise over 30 minutes. After warming to room temperature, the clear solution was stirred for 2 hours, then poured into a mixture of saturated aqueous sodium hydrogen carbonate (200 ml) andpH 6.86 standard buffer (200 ml).
Organic solvent was removed by evaporation, and the remaining WO 01/60846 PCT/JP01/01204 155 aqueous solution purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting with aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (5.53 g).
NMR (DMSO-d 6
D
2 0, 0.97 (3H, d, J=6.64Hz), 1.15 (3H, d, J=5.52Hz), 1.30-1.70 (3H, 1.80-2.50 (6H, m), 2.70-4.00 (14H, 4.20-4.60 (8H, 4.70-4.90 (2H, 5.10-5.40 (2H, 5.70-6.10 (1H, 6.70-6.90 (2H, 7.06 (1H, s) ESI MASS (m/z)(Positive): 997.3 (M++Na) Example 7 A solution of the starting compound (0.5 g) in dimethylformamide (10 ml) was treated with 4-methylcyclohexyl)piperazinyl]phenyl]-1,3,4-thiadiazol-2yl]benzoic acid benzotriazol-1-yl ester (0.3 g) and diisopropylethylamine (0.13 ml) and stirred for 20 hours at room temperature. Ethyl acetate (100 ml) was added and the resulting precipitate collected, washed with ethyl acetate, and dried to give the object compound (0.5 g).
NMR (DMSO-d 6 0.90 (3H, d, J=6.8Hz), 0.97 (3H, d, J=6.6Hz), 1.13 (3H, d, J=5.0Hz), 1.43-6.10 (78H, m), 6.69-8.72 (18H, m) ESI MASS (m/z)(Negative): 1418.4 (M Example 8 To a suspension of the starting compound (0.38 g) in a mixture of methanol (7.6 ml) and tetrahydrofuran (1.9 ml) were successively added triphenylphosphine (0.04 g), tetrakis(triphenylphosphine)palladium(0) (0.088 g) and morpholine (0.14 ml) with stirring and the mixture was stirred at ambient temperature for 15 hours. To the reaction mixture WO 01/60846 PCT/JP01/01204 156 was added ethyl acetate (100 ml) The resulting precipitate was collected by filtration and dried in vacuo. The precipitate was dissolved in a mixture of water and 1N sodium hydroxide and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (100 ml) eluting with 40% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (0.25 g).
NMR (DMSO-d 6 0.90 (3H, d, J=6.7Hz), 0.98 (3H, d, J=6.7Hz), 1.11 (3H, d, J=5.7Hz), 1.42-5.23 (56H, m), 6.69-8.92 (17H, m) ESI MASS (m/z)(Negative): 1334.4 (M Elemental Analysis Calcd. for C 61
H
82
N
1 2 0 18
S
2 -8H 2 0: C 49.52, H 6.68, N 11.36 Found C 49.25, H 6.41, N 11.20 Example 9 The suspension of a mixture of the starting compound (9) (100 mg), 1,3-dihydroxyacetate (13.5 mg) and acetic acid (0.13 ml) in a mixture of methanol (1.5 ml) and dimethylformamide (0.7 ml) was added sodium cyanoborohydride (9.4 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight. To the reaction mixture was added ethyl acetate (20 ml). The resulting precipitate was collected by filtration and dried in vacuo. The precipitate was dissolved in a mixture of water and lN sodium hydroxide and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) ml) eluting with 40% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (55 mg).
NMR (DMSO-d 6 0.90 (3H, d, J=6.8Hz), 0.98 (3H, d, WO 01/60846 PCT/JP01/01204 157 J=6.7Hz), 1.11 (3H, d, J=5.5Hz), 1.43-5.24 (62H, m), 6.69-8.85 (17H, m) ESI MASS (m/z)(Negative): 1408.3(M Example To a solution of a mixture of the starting compound 1,3-dihydroxyacetone (1.19 g) and acetic acid (1.14 ml) in a mixture of methanol (120 ml) and dimethylformamide (55 ml) was added sodium cyanoborohydride (835 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight. To a reaction mixture was poured into ethyl acetate (700 ml). The resulting precipitates were collected by filtration, washed with ethyl acetate (100 ml) and dried in vacuo. The precipitates were dissolved in a mixture of 30% aqueous acetonitrile (800 ml) and 1N sodium hydroxide ml) The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (440 ml) eluting in turn with water and aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (10) (5.22 g).
IR (KBr): 1632, 1535, 1518, 1443, 1269, 1082, 1047 cm 1 NMR (DMSO-d 6
D
2 0, 0.82 (3H, d, J=6.83Hz),0.97 (3H, d, J=6.81Hz), 1.02 (3H, d, J=6.18Hz), 1.24 (26H, s), 1.35-2.45 (14H, 2.75-3.40 (5H, 3.60-4.50 4.7-4.90 (2H, 6.65-6.80 (2H, 7.01 (1H, s) ESI MASS (m/z)(Positive): 1088.4 (M4+Na) Example 11 To a solution of the starting compound (11) (4.0 g) in dimethylformamide (40 ml) were successively added diisopropylethylamine (1.45 ml) and 9-fluorenylmethyl chloroformate (1.03 and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was poured into WO 01/60846 PCT/JP01/01204 158 water (200 ml). The solution was purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting in turn with a mixture of saturated aqueous sodium chloride (400 ml), saturated aqueous sodium hydrogen carbonate (50 ml) and water (400 ml), and aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (11) (2.82 g).
IR (KBr) 1666, 1632, 1518, 1446, 1273, 1246, 1082, 1047 cm-1 cm NMR (DMSO-d 6
D
2 0, 0.80-1.10 (9H, 1.23 (26H, s), 1.35-2.45 (12H, 2.60-3.40 (6H, 3.60-4.55 (18H, 4.65-4.90 (2H, 6.65-6.85 (2H, 6.97 (1H, 7.30-7.50 (4H, 7.60-7.95 (4H, m) ESI MASS (m/z)(Negative): 1423.7 (M+-Na) Elemental Analysis Calcd. for C 69
H
99 NgO 22 SNa-6H 2 0: C 53.27, H 7.19, N 7.20 Found C 53.45, H 7.21, N 7.10 Example 12 To a solution of the object compound (12) (1.21 g) in dimethylformamide (15 ml) were successively added diisopropylethylamine (0.26 ml) and di-tert-butyl dicarbonate (285 mg), and the mixture was stirred at ambient temperature overnight. The reaction mixture was poured into a mixture of pH 6.86 standard buffer solution (150 ml), saturated aqueous sodium chloride (50 ml) and saturated aqueous sodium hydrogen carbonate (20 ml). The mixture was purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting with aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (12) (1.19 g).
WO 01/60846 PCT/JP01/01204 159 IR (KBr): 1662, 1632, 1535, 1518, 1444, 1367, 1272, -i 1250 cm 1 NMR (DMSO-d 6
D
2 0, 0.85 (3H, d, J=6.76Hz), 0.96 (3H, d, J=6.77Hz), 1.04 (3H, d, J=5.50Hz), 1.23 (26H, 1.37 (9H, 1.40-1.50 (2H, 1.55-2.50 (10H, m), 2.80-3.40 (6H, 3.50-4.45 (14H, 6.65-6.80 (2H, 6.96 (1H, s) ESI MASS (m/z)(Negative): 1301.6 (M+-Na) Example 13 To a solution of a mixture of starting compound (13) (1.62 g) and diisopropylethylamine (0.58 ml) in DMF (16 ml) was added 9-fluorenylmethyloxycarbonyl chloride (493 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature for 3 hours. The reaction mixture was poured into ethyl acetate (250 ml). To the mixtures was added pH 6.86 standard buffer solution (100 ml) and 5% aqueous sodium chloride ml), and the aqueous layer was separated. The organic layer was extracted with 5% aqueous sodium chloride (100 ml), and these aqueous layers were collected and evaporated in vacuo to remove organic solvent. The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) eluting with acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (13) (1.38 g).
NMR (DMSO-d 6
D
2 0, 0.89 (3H, d, J=6.26Hz), 1.09 (3H, broad 1.33 (9H, 1.40-2.10 (5H, m), 2.10-2.35 (2H, 2.75-3.40 (5H, 3.50-4.50 (16H, 4.60-4.90 (2H, 6.65-6.80 (2H, 6.97 (1H, 7.25-7.50 (4H, 7.70 (2H, d, J=6.82Hz), 7.88 (2H, d, J=6.77Hz) ESI MASS (m/z)(Positive): 1331.3 (M++Na) Elemental Analysis Calcd. for C 58
H
77
N
8 0 23 SNa-4H 2 0: WO 01/60846 PCT/JP01/01204 160 C 50.43, H 6.20, N 8.11 Found: C 50.14, H 6.28, N 8.12 Example 14 To a solution of a mixture of starting compound (14) (300 mg), 2-oxo-1,3-diacetoxypropane (121 mg) and acetic acid (40 ~1) in a mixture of methanol (4.0 ml) and DMF (4.0ml) was added sodium cyanoborohydride (29 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight.
The reaction mixture was concentrated in vacuo. To the resulting residue was added pH 6.86 standard buffer solution (10 ml) and acetonitrile (2 ml), and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (80 ml) eluting with acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (14) (60 mg).
NMR (DMSO-d 6
D
2 0, 0.98 (3H, d, J=6.83Hz), 1.07 (3H, d, J=5.34Hz), 1.20-1.60 (10H, 1.60-1.90 (5H, m), 1.96 (3H, 2.01 (3H, 3.20 (3H, 3.31 (4H, t, J=6.33Hz), 3.80-4.55 (16H, 4.75-4.90 (2H, m), 6.65-6.80 (2H, 7.03 (1H, 7.14 (2H, d, J=8.84Hz), 7.90-8.15 (6H, m) ESI MASS (m/z)(Negative): 1455.3 Example To a solution of starting compound (15) (58 mg) in a mixture of methanol (3 ml) and water (3 ml) were added morpholine and saturated aqueous sodium carbonate (1 ml), and the mixture was stirred at ambient temperature for 1 hour. The reaction mixture was poured into pH 6.86 standard buffer solution (60 ml), and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso WO 01/60846 PCT/JP01/01204 161 Co., Ltd.)) (50 ml) eluting with 30% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (15) (35 mg).
NMR (DMSO-d 6
D
2 0, 0.97 (3H, d, J=6.78Hz), 1.12 (3H, broad 1.25-1.65 (8H, 1.65-2.00 (4H, m), 2.01 (3H, d, 3.21 (3H, 3.31 (4H, t, J=6.34Hz), 3.70-4.50 (14H, 4.85-4.90 (2H, 6.60-6.95 (2H, 7.00 (1H, 7.14 (2H, d, J=8.74Hz), 8.00 (2H, d, J=8.77Hz), 8.03 (2H, d, J=7.63Hz), 8.12 (2H, d, J=8.42Hz) ESI MASS (m/z)(Negative): 1413.4 (M+-l-Na) Example 16 To a solution of starting compound (16) (100 mg) in DMF (3 ml) were added 4-[5-[4-(6-methoxyhexyl)phenyl][1,3,4]thiadiazol-2-yl]benzoic acid benzotriazol-1-yl ester (71 mg) and diisopropylethylamine (23 il) with stirring, and the mixture was stirred at ambient temperature overnight. To the reaction mixture was added ethyl acetate (30 ml). The resulting precipitates were collected by filtration, washed with ethyl acetate (10 ml) and dried in vacuo. The resulting residue was dissolved in a mixture of pH 6.86 standard buffer solution and IN sodium hydroxide, and insoluble materials were filtered off and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (50 ml) eluting with 30% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (16) (86.5 mg).
NMR (DMSO-d 6
D
2 0, 0.97 (3H, d, J-6.53Hz), 1.08 (3H, d, J=8.66Hz), 1.30-2.00 (14H, 2.80-3.10 (4H, 3.22 (3H, 3.90-4.55 (16H, 4.65-4.90 (2H, 5.10-5.40 (2H, 6.82 (2H, br 7.00 (1H, s), 7.14 (2H, d, J=9.17Hz), 7.90-8.20 (6H, m) WO 01/60846 PCT/JP01/01204 162 ESI MASS (m/z)(Negative): 1441.4 (M+-1-Na) Example 17 To a solution of starting compound (17) (200 mg) in N,N-dimethylformamide (DMF) (3 ml) were added 4'-[4-4-(cis- 2,6-dimethylmorpholin-4-yl)phenyl]piperazin-l-yl]-1,1'biphenyl-4-carboxylic acid benzotriazol-1-yl ester (57 mg) and diisopropylethylamine (22 il) with stirring, and the mixture was stirred at ambient temperature overnight. To the reaction mixture was added ethyl acetate (30 ml). The resulting precipitates were collected by filtration, washed with ethyl acetate (10 ml) and dried in vacuo. The resulting residue was dissolved in a mixture of pH 6.86 standard buffer solution and lN sodium hydroxide, and insoluble material were filtered off and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (100 ml) eluting with 40% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (17) (230 mg).
NMR (DMSO-d 6
D
2 0, 0.97 (3H; d, J=6.B2Hz), 1.14 (6H, d, J=6.16Hz), 1.25 (3H, d, J=6.34Hz), 1.30-2.40 (6H, 3.00-3.40 (10H, 3.60-4.10 (10H, m), 4.10-4.55 (6H, 4.60-4.80 (4H, 5.05-5.50 (4H, 5.80-6.10 (2H, 6.80-7.00 (4H, 7.08 (2H, d, J=8.10Hz), 7.11 (2H, d, J=8.88Hz), 7.42 (1H, s), 7.66 (2H, d, J=8.64Hz), 7.72 (2H, d, J=8.46Hz), 7.93 (2H, d, J=8.38Hz) ESI MASS (m/z)(Negative): 1584.6 (M+-Na) Example 18 A mixture of 4-[5-[4-(6-methoxyhexyloxy)phenyl]isoxazol-3-yl]benzoic acid (70 mg), l-hydroxybenzotriazole (35.8 mg), l-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (40.6 mg) and N,N-diisopropylethylamine (46.1 l) WO 01/60846 PCT/JP01/01204 163 in N,N-dimethylformamide (2 ml) was stirred for 3 hours. To the reaction mixture was added starting compound (18) (200 mg) and the resulting mixture was stirred for 19 hours. To the reaction mixture was added ethyl acetate (100 ml). The resulting precipitate was collected by filtration and washed with diisopropyl ether to give object compound (18) as a crude white powder (294.4 mg), that was used crude in the next reaction.
The following compound was obtained according to a similar manner to that of Example 18.
Example 19 The object compound (19) was used directly in the next reaction without purification.
Example To a solution of starting compound (20) (287.9 mg) in N,N-dimethylformamide (3 ml) was added piperidine (0.17 ml) at room temperature. The solution was stirred for 1 hour at the same temperature. Ethyl acetate was added to the reaction mixture. The powder was collected by filtration to give crude material (203.8 mg). The crude material was purified by column chromatography on ODS to give object compound (20) (85.6 mg).
IR (KBr): 1632, 1512, 1446, 1230 cm 1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.08 (3H, d, J=5.2Hz), 1.5-3.0 (23H, 3.0-4.5 (39H, 4.6- 5.4 (10H, 6.6-7.1 (11H, 7.17 (2H, d, J=8.7Hz), 7.3-7.6 (2H, 7.81 (2H, d, J=8.6Hz), 8.0-8.5 (2H, 8.71 (1H, s) MASS 1488 The following compound was obtained according to a similar manner to that of Example Example 21 WO 01/60846 PCT/JP01/01204 164 IR (KBr): 1632, 1512, 1444, 1232 cm-1 NMR (DMSO-d 6 0.97 (3H, d, J=6.8Hz), 1.08 (3H, d, 1.2-3.0 (28H, 3.0-4.5 (38H, 4.6- 5.4 (10H, 6.6-7.1 (9H, 7.3-7.7 (2H, 7.7-8.0 (3H, 8.0-8.5 (5H, 8.71 (1H, s) MASS 1456 Example 22 To a solution of starting compound (22) (0.22 g) in a mixture of methanol (4 ml) and THF (1 ml) were successively added triphenylphosphine (14 mg), tetrakis(triphenylphosphine)palladium(0) (8 mg) and morpholine gl) with stirring and the mixture was stirred at ambient temperature for 3 hours. The reaction mixture was concentrated in vacuo. The resulting residue was dissolved in a mixture of pH 6.86 standard buffer solution and IN sodium hydroxide, insoluble materials were filtered off and the solution was subjected to column chromatography on ODS (Daiso-gel, SP- 120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (100 ml) eluting with 30% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (22) (85 mg).
IR (KBr): 1633, 1537, 1516, 1450, 1234 cm-1 NMR (DMSO-d 6
D
2 0, 0.98 (3H, d, J=7.09Hz), 1.05 (3H, d, J=7.00Hz), 1.15 (6H, d, J=6.21Hz)i 1.60-2.30 (8H, 2.75-3.45 (14H, 3.80-4.50 (10H, 4.81 (1H, br 6.65-7.20 (8H, 7.50-7.80 (5H, 7.94 (2H, d, J=8.49Hz) ESI MASS (m/z)(Negative): 1416.4 (M +1) Elemental Analysis Calcd. for C 6 7
H
9 1
N
1 1 0 2 1 S7H 2 0: C 52.10, H 6.85, N 9.97 Found: C 52.29, H 6.60, N 9.61 The following compounds [Examples 23 to 32] were obtained WO 01/60846 PCT/JP01/01204 165 according to a similar manner to that of Example 22.
Example 23 NMR (DMSO-d 6
D
2 0, 0.97 (3H, d, J=6.84Hz), 1.11 (3H, d, J=5.43Hz), 1.30-1.90 (14H, 2.80-3.20 (5H, m), 3.22 (3H, 3.31 (2H, t, J=6.16Hz), 3.80-4.20 (6H, 4.26 (2H, broad 4.30-4.50 (3H, 4.70-4.90 (1H, 6.72 (1H, d, J=8.14Hz), 6.78 (1H, d, J=10.5Hz), 7.01 (1H, 7.14 (2H, d, J=8.70Hz), 7.98 (2H, d, J=8.90Hz), 8.05 (2H, d, J=8.68Hz), 8.12 (2H, d, J=8.68Hz) MASS (m/z)(Negative): 1357.5 (M -1) Example 24 IR (KBr): 2933, 1633, 1531, 1518, 1444, 1419, 1385, 1346 cm 1 NMR (DMSO-d 6 0.90 (3H, d, J-6.7Hz), 0.98 (3H, d, J=6.7Hz), 1.12 (3H, d, J=5.5Hz), 1.32-2.68 (23H, m), 2.82-2.98 (2H, 3.07-4.54 (25H, 4.74-5.50 6.70 (1H, d, J=8.1Hz), 6.78 (1H, d, J=8.1Hz), 7.00 (1H, 7.09 (2H, d, J=9.0Hz), 7.36-7.70 (2H, 7.86 (2H, d, J=8.8Hz), 8.00-8.50 (6H, 8.71 (1H, s), 8.82-8.97 (1H, m) ESI MASS 1407.5 (M +1) Elemental Analysis Calcd. for C 6 4
H
88
N
1 2 0 2 0
S
2 -7H 2 0: C 50.06, H 6.69, N 10.94 Found: C 49.99, H 6.76, N 10.73 Example IR (KBr): 3353.6, 1666.2, 1648.8, 1631.5, 1540.8, 1508.1, 1452.1, 1436.7, 1257.4 cm-1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.11 (3H, d, 1.2-5.6 (59H, 6.71 (1H, d, J=8.2Hz), 6.78 (1H, d, J=9.6Hz), 7.00 (1H, 7.12 (2H, d, J=8.8Hz), 7.44 (1H, d, J=8.5Hz), 7.55 (1H, 7.85 WO 01/60846 PCT/JP01/01204 166 (2H, d, J=8.6Hz), 7.99 (2H, d, J=8.5Hz), 8.05 (2H, d, J=8.6Hz), 7.3-8.5 (3H, 8.71 (1H, 8.7-9.0 (1H, m) MASS 1340.4 (M--Na) Elemental Analysis Calcd. for C 6 1
H
83
N
9
O
2 3 S-6H 2 0: C 50.51, H 6.60, N 8.69 Found: C 50.67, H 6.60, N 8.62 Example 26 IR (KBr): 3380.6, 1675.8, 1648.8, 1621.8, 1540.8, 1506.1, 1454.1, 1434.8, 1257.4 cm-1 NMR (DMSO-d 6 0.98 (3H, d, J=6.8Hz), 1.03 (6H, d, J=6.3Hz), 1.12 (3H, d, J=5.5Hz), 1.2-5.6 (64H, m), 6.71 (1H, d, J=8.1Hz), 6.77 (1H, d, J=9.4Hz), 7.00 (1H, 7.12 (2H, d, J=8.9Hz), 7.43 (1H, d, J=7.7Hz), 7.55 (1H, 7.85 (2H, d, J=8.6Hz), 7.99 (2H, d, J=8.8Hz), 8.05 (2H, d, J-8.8Hz), 7.3-8.5 (3H, 8.71 (1H, s), 8.82 (1H, d, J=5.7Hz) MASS 1437.4 Elemental Analysis Calcd. for C 6 7
H
94
N
1 0 0 2 3 S-6H 2 0: C 52.00, H 6.90, N 9.05 Found: C 51.91, H 6.91, N 8.77 Example 27 IR (KBr): 2931, 2854, 1632, 1510, 1446, 1385, 1325 cm 1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.12 (3H, d, 1.08-2.62 (23H, 2.62-4.50 (37H, m), 4.66-5.45 (10H, 6.70 (1H, d, J=8.1Hz), 6.78 (1H, d, J=8.1Hz), 6.83-7.09 (7H, 7.34-8.00 (3H, m), 7.80 (2H, d, J=8.7Hz), 8.00-8.49 (2H, 8.71 (1H, s) MASS 1408.4 (M +1) Elemental Analysis Calcd. for C 6 6
H
95
N
11
O
21 S-7H 2 0: C 51.59, H 7.15, N 10.03 Found: C 51.77, H 7.05, N 9.82 WO 01/60846 PCT/JP01/01204 167 Example 28 IR (KBr): 2974, 2937, 1633, 1533, 1512, 1444, 1383, 1327 cmr 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.11 (3H, d, J=5.2Hz), 1.18 (6H, d, J=6.1Hz), 1.59-2.65 (11H, m), 2.65-4.56 (27H, 4.70-5.36 (10H, 6.71 (1H, d, J=8.1Hz), 6.78 (1H, d, J=8.1Hz), 7.00 (1H, 7.08 (2H, d, J=8.8Hz), 7.38-7.99 (3H, 7.68 (2H, d, J=8.7Hz), 7.86 (2H, d, J=8.5Hz), 8.00-8.46 (7H, m), 8.71 (1H, 8.80-8.95 (1H, m) MASS 1440.3 (M++Na) Elemental Analysis Calcd. for C 6 5
H
85
N
11
O
21
S
2 -8H20: C 49.96, H 6.39, N 9.86 Found: C 50.03, H 6.17, N 9.47 Example 29 IR (KBr): 3386.4, 1633.4, 1502.3, 1446.4, 1232.3 cm 1 NMR (DMSO-d 6 0.96 (3H, d, J=6.6Hz), 1.0-1.3 (9H, 1.3-5.6 (57H, 6.70 (1H, d, J=8.1Hz), 6.77 (1H, d, J=9.7Hz), 6.9-7.2 (7H, 7.3-9.0 (13H, m) MASS 1416.4 (M--Na) Example IR (KBr): 3365.2, 1631.5, 1517.7, 1465.6, 1444.4, 1257.4 cm 1 MASS 1368.3 Elemental Analysis Calcd. for C 60
H
79
N
1 1 0 2 2
S
2 '7H 2 0: C 48.15, H 6.26, N 10.30 Found: C 48.26, H 6.17, N 10.35 Example 31 IR (KBr): 3458, 3425, 3398, 3386, 3363, 2935, 1635, 1523, 1462, 1244 cm- 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.12 (3H, d, WO 01/60846 PCT/JPOI/01204 168 6Hz) 1. 20-1. 60 (12H, in) 1.70-2. 45 (12H-, in), 2.80-3.20 (9H, mn), 3.21 (3H, 3.40-4.60 (24H, in), 4.70-5.40 (12H, in), 6.71 (1H, di, J=8.lHz), 6.60-6.80 (1H, mn), 7.00 (lH, di, J=l.4Hz), 7.08 (2H, di, J=9Hz), 7.35-7.65 (2H, in), 7.75 (2H1, d, J=8.8Hz), 7.80-8.10 in), 8.20-8.40 (TH, mn), 8.60-8.80 (2H, mn), 8.80 (1H, s) MASS (AEC-ES-Negative): 1497 (M+-llNa) Elemental Analysis Calcd. for C 6 7
H
9 lNl 2
O
2 lS 3 -E-1/2H 2
O:
C 48.75, M 6.55, N 10.18 Found: C 48.52, H 6.47, N 9.74 Example 32 IR (KBr) 3464, 3425, 3398, 3386, 3363, 2940, 1635, 1523, 1461 cm-1 NMR (DMSO-d 6 0.97 (3H, di, J=6.8Hz), 1.12 (3H, d, J=5.6Hz), 1.40-1-60 (6H, in), 1.65-2.45 (9H, in), 2.60-3.20 in), 3-21 (3H1, 3.40-3.80 (15H1, m), 3.80-4.60 (14H-, mn), 4.65-5.50 (9H1, mn), 6.71 (1H1, d, J=r.lHz), 6.75 (1H1, dd, J=1.6 and 8.3Hz), 7.03 (1H, d, c=l.6Hz), 7.09 (2H, d, J =9Hz), 7.40-7.65 (2H, mn), 7.75 (2H, d, J=8.8Hz), 7.80-8.00 (4H, in), 8.18-8.30 (1H, in), 8.55-8.70 (1H, in), 8.75 (2H, di, J=8.7Hz) MASS (in/z) (API-ES-Negative) 1453 Elemental Analysis Caled. for C 6 5
H
38 Nl 2
O
2 2
S
2 -6H 2
O:
C 49.27, H 6.25, N 10.61 Found: C 49.03, H 6.33, N 10.30 Example 33 To a solution of starting compound (33) (12.50 g) and diisopropylethylanine (3.67 ml) in N,N-dimethylfornamide (250 ml) was added 4-[2-L4-(4-inethoxybutoxy)phenyllimidazor2,1-bl [1,3,4lthiadiazol-6-yllbenizoic acid benzotriazol-l-yl ester at room temperature. The solution was stirred for 4 hours at the same temperature, during which period additional 4- WO 01/60846 PCT/JP01/01204 169 [2-[4-(4-methoxybutoxy)phenyl]imidazo[2,1-b][1,3,4]thiadiazol-6-yl]benzoic acid benzotriazol-l-yl ester was added to the mixture. The reaction mixture was then filtered. To the filtrate was added piperidine (9.33 ml) at room temperature. The solution was stirred for 1 hour at the same temperature. Ethyl acetate was added to the reaction mixture. The powder was collected by filtration to give crude material (16.12 The crude material was purified by column chromatography on ODS to give object compound (11.10 g).
IR (KBr): 1659, 1633, 1529, 1518, 1466, 1444, 1255 cm 1 NMR (DMSO-d 6 6) 0.98 (3H, d, J=6.7Hz), 1.00 (3H, d, J=5.8Hz), 1.5-2.6 (12H, 2.8-3.6 (33H, 4.7- 5.4 (10H, 6.65-6.85 (2H, 7.00 (1H, 7.15 (2H, d, J=8.9Hz), 7.3-7.7 (2H, 7.90 (2H, d, J=8.8Hz), 7.96 (4H, 8.0-8.5 (2H, 8.71 (1H, s), 8.85 (1H, s) MASS 1392 (M++23) Elemental Analysis Calcd. for C 6 0
H
79
N
11 0 2 2
S
2 -5H 2 0: C 49.34, H 6.14, N 10.55 Found: C 49.30, H 6.23, N 10.53 The following compounds [Examples 34 and 44] were obtained according to a similar manner to that of Example 33.
Example 34 IR (KBr): 3463, 3423, 3359, 2941, 2883, 1633, 1614, 1523, 1462 cm 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.10 (3H, d, J=5.6Hz), 1.35-2.20 (10H, 2.80-3.20 (2H, 3.22 (3H, 3.30-3.80 (10H, 3.80-4.60 (10H, m), 4.70-5.35 (9H, 6.71 (1H, d, J=8.1Hz), 6.65-6.90 (1H, 7.00 (1H, br 7.09 (2H, d, J=9Hz), 7.40-7.70 (2H, 7.43 (2H, d, J=8.6Hz), 7.80-8.00 (4H, 8.10-8.50 (2H, 8.60-8.80 (3H, m) MASS (API-ES-Negative): 1440 WO 01/60846 PCT/JP01/01204 170 Elemental Analysis Calcd. for C 64 H8 6
N
1 2 0 2 2
S
2 -6-1/2H 2 0: C 49.36, H 6.36, N 10.80 Found: C 49.20, H 6.50, N 10.66 Example NMR (DMSO-d 6 0.90 (3H, d, J=6.7Hz), 0.98 (3H, d, J=6.8Hz), 1.11 (3H, d, J=5.7Hz), 1.43-5.24 (62H, m), 6.69-8.85 (17H, m) MASS 1408.5 Example 36 MASS 1491.4 (M+-HN+Et(iPr) 2 Example 37 MASS 1576.5 (M+-HN+Et(iPr) 2 Example 38 MASS 1584.4 (M+-HN Et(iPr) 2 Example 39 The object compound (39) reaction without purification.
Example The object compound (40) reaction without purification.
Example 41 The object compound (41) reaction without purification.
was used directly in the next was used directly in the next was used directly in the next Example 42 The object compound (42) was used directly in the next reaction without purification.
WO 01/60846 PCT/JP01/01204 171 The following compounds [Examples 43 and 44] were obtained according to a similar manner to that of Example Example 43 NMR (DMSO-d 6
D
2 0, 0.89 (3H, d, J=6.22Hz), 1.14 (3H, br 1.35-2.40 (6H, 2.65-3.00 (1H, m), 3.60-4.50 (14H, 4.55-4.80 (2H, 5.28 (1H, s), 6.65-6.80 (2H, 6.98 (1H, 7.20-7.50 (4H, m), 7.69 (2H, d, J=7.08Hz), 7.84 (2H, d, J=7.27Hz) ESI MASS (m/z)(Negative): 1185.4 Example 44 NMR (DMSO-d 6
D
2 0, 0.95 (3H, d, J=6.77Hz), 1.12 (3H, d, J=4.94Hz), 1.20-1.75 (4H, 1.80-2.50 (4H, m), 2.65-2.90 (1H, 3.00-3.40 (4H, 3.60-4.05 (6H, 4.17 (2H, J=7.17Hz), 4.25-4.90 (7H, 5.05-5.35 (2H, 5.75-6.10 (1H, 6.65-6.85 (2H, 6.97 (1H, s) ESI MASS (m/z)(Positive): 1048.3 (M Example To a solution of a mixture of starting compound (45) 1,3-dihydroxyacetone (364 mg) and acetic acid (0.58 ml) in a mixture of methanol (30 ml) and DMF (14 ml) was added sodium cyanoborohydride (258 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight.
To the reaction mixture was added ethyl acetate (200 ml). The resulting precipitates were collected by filtration and dried in vacuo. The precipitates were dissolved in a mixture of pH 6.86 standard buffer solution (100 ml) and acetonitrile (20 ml), and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) eluting with 15% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The WO 01/60846 PCT/JP01/01204 172 residue was lyophilized to give object compound (45) (1.63 g).
NMR (DMSO-d 6
D
2 0, 0.98 (3H, d, J=6.75Hz), 1.08 (3H, d, J=5.69Hz), 1.35 (9H, 1.45-2.05 (5H, m), 2.15-2.50 (4H, 2.70-3.35 (7H, 3.50-4.50 (16H, 4.70-4.90 (2H, 6.71 (1H, d, J=8.13Hz), 6.78 (1H, d, J=9.91Hz), 7.01 (1H, s) ESI MASS (m/z)(Positive): 1088.4(M++Na) The following compounds [Examples 46 to 52] were obtained according to a similar manner to that of Example Example 46 IR (KBr): 3353.6, 1635.3, 1444.4, 1257.4, 1085.7, 1047.2 cm 1 NMR (DMSO-d 6 0.97 (3H, d, J=6.6Hz), 1.14 (3H, d, J=5.4Hz), 1.2-5.6 (61H, 6.71 (1H, d, 6.77 (1H, d, J=10.3Hz), 6.96 7.13 (2H, d, J=8.8Hz), 7.97 (2H, d, J=8.7Hz), 8.08 (4H, 7.4-8.9 (6H, m) MASS 1371.4 (M-1) Example 47 IR (KBr): 3353.6, 1635.3, 1531.2, 1517.7, 1444.4, 1257.4, 1087.7, 1045.2 cm-1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.0-5.6 (64H, 6.6-6.8 (2H, 6.99 (1H, 7.14 (2H, d, J=8.9Hz), 7.97 (2H, d, J=8.8Hz), 8.08 (4H, 7.3-9.0 (6H, m) MASS 1371.3 Elemental Analysis Calcd. for C 61
H
8 4
N
1 00 2 2
S
2 -7H20: C 48.86, H 6.59, N 9.34 Found: C 49.00, H 6.39, N 9.24 Example 48 IR (KBr): 3384.5, 1658.5, 1635.3, 1529.3, 1517.7, WO 01/60846 PCT/JPO/01204 173 1446.4, 1257.4, 1085.7, 1045.2 cm 1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.13 (3H, d, 1.2-5.3 (65H, in), 6.91 (1H, d, J=8.1Hz), 6.77 (1H, d, J=9.9Hz), 6.97 (1H, 7.13 (2H, d, J=8.9Hz), 7.97 (2H, d, J=8.8Hz), 8.09 (4H, 7.4-8.9 (6H, m) MASS 1431.3 (M-1I) Elemental Analysis Calcd. for C 6 3
H
8 6
N
10
O
24
S
2 '8H 2 0: C 47.96, H 6.64, N 8.88 Found: C 48.21, H 6.35, N 8.87 Example 49 IR (KBr): 3371.0, 1648.8, 1631.5, 1538.9, 1513.8, 1442.5, 1257.4, 1083.8, 1045.2 cm- 1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.10 (3H, d, 1.2-5.4 (65H, 6.71 (11, d, J=8.2Hz), 6.77 (1H, cl, J=10.2Hz), 6.99 (1H, 7.14 (2H, d, J=8.7Hz), 7.97 (2H, d, J=87Hz), 8.09 (4H, 7.3-9.0 (6H, m) MASS 1401.3 Elemental Analysis Calcd. for C 6 2
H
8 6
N
10 0 23
S
2 -71 2 0: C 48.68, H 6.59, N 9.16 Found: C 48.83, H 6.39, N 9.13 Exam]le IR (KBr): 3350, 2933, 2862, 1658.5, 1635, 1516, 1444, 1257, 1084, 1043 cm- 1 NNR (DMSO-d 6
D
2 0, 0.98 (3H, d, J=6.7Hz), 1.09 (3H, d, J=5.8Hz), 1.2-4.8 (49H, complex 3.21 (3H, s), 3.31 (2H, t, J=6.4Hz), 6.8-6.9 (2H, 7.02 (1H, br 7.15 (2H, d, J=8.9Hz), 7.98 (2H, d, J=8.9Hz), 8.10 (4H, s) MASS 1485.4 (M++Na) Elemental Analysis Calod. for C 64
H
90
N
10 0 25
S
2 -6H 2 0: C 48.91, H 6.54, N 8.91 WO 01/60846 PCT/JPOI/01204 174 Found: C 49.18, H 6.55, N 8.90 Example 51 NMR (DMSO-d 6 0.86 (3H, d, J=6.3Hz), 0.98 (3H, d, J=6.8Hz), 1.11 (3H, d, J=5.7Hz), 1.21-5.24 (62H, mn), 6-69-8.89 (17H, m) MASS (in/z) 1408.5, 1407.4 Elemental Analysis Calod. for C 6 4
H
8 8
N
1 2 0 2 0
S
2 -7H 2 0: C 50.06, H 6.69, N 10.94 Found: C 49.96, H 6.86, N 10.82 Examiple 52 IR 1633, 1606, 1529, 1516, 1466 crrJ' NMR (DMSO-d 6 0.96 (3H, d, J=6.7Hz), 1.11 (3H, d, J=5.7Hz), 1.2-2.6 (18H, mn), 2.8-4.6 (39H, mn), 4.7- 5.4 (9H, in), 6.7-6.9 (2H, mn), 7.0-7.2 (3H, in), 7.3-7.6 (2H, in), 7.75 (2H, d, J=8.7z), 7.7-8.0 (5H, in), 8.2-8.5 (lH, in), 8.6-8.75 (1H, in), 8.80 8.85 (lH, s) MASS (in/z) 1481 Elemental Analysis Calcd. for C 6 6
H
9 0
N
1 2 0 2 3
S
2 *7H 2 0: C 49.25, H 6.51, N 10.44 Found: C 49.30, h 6.34, N 10.40 The following compound was obtained according to a similar manner to that of Example 1.
Example 53 IR (KBr): 2937.1, 1651, 1631.5, 1539, 1523.5 cm- 1 MASS (in/z) 1293.3 Examp2le 54 To asolution of starting compound (54) (300 itt) in methanol (12 ml) was added 10% hydrochiolic acid in methanol (6 ml) at room temperature. The solution was stirred for 3 hours at the WO 01/60846 PCT/JP01/01204 175 same temperature. The solvent was evaporated under reduced pressure to remove hydrochloric acid and methanol. To the residue was added water and the mixture was lyophilized. The residue was purified by column chromatography on ODS to give object compound (54) (119 mg).
IR (KBr): 1649, 1633, 1608, 1539, 1525 cm 1 MASS 1351 (M++23) Elemental Analysis Calcd. for C 6 4
H
88
N
12 0 17 S8H 2 0: C 52.16, H 7.11, N 11.41 Found: C 52.13, H 7.05, N 11.36 The following compounds [Example 55 to 71] were obtained according to a similar manner to that of Example 33.
Example IR (KBr): 3358, 1633, 1608, 1535, 1516, 1443, 1419, 1271, 1238 cm 1 NNR (DMSO-d 6
+D
2 0, 0.89 (6H, 0.98 (3H, d, J=6.7Hz), 1.10 (3H, d, J=5.8Hz), 1.1-2.6 (20H,m), 2.6-4.5 (29H, 4.7-4.9 (2H, 6.7-6.9 (2H, 7.0-7.1 (1H, m), 7.08 (2H, d, J=9.2Hz), 7.86 (2H, d, J=8.5Hz), 8.2 (4H, m) ESI MASS (Negative): 1422.3 Elemental Analysis Calcd. for C 65
H
90
N
1 2
O
2 0
S
2 7.5H 2 0: C 50.09, H 6.79, N 10.78 Found: C 49.94, H 6.59, N 10.52 Example 56 IR (KBr): 3462, 3458, 3425, 3399, 3367, 1633, 1578, 1440 cm 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.7Hz), 1.11 (3H, d, J=5.6Hz), 1.20-1.50 (3H, 1.60-2.40 (18H, m), 2.50-2.70 (4H, 2.75-3.20 (7H, 3.4C-3.60 (6H, 3.70-4.50 (14H, 4.62 (2H, br 4.65-4.80 (3H, 4.80-5.40 (8H, 6.60-6.80 (2H, 7.00 (1H, WO 01/60846 PCT/JPOI/01204 176 br s) 7.07 (2H, d, 9Hz) 7. 40-7. 60 (2H, in), '7.85 (2H, d, J=8.7Hz), 7.90-8.20 (4H, mn), 8.2D-8.40 (1H, mn), 8.71 (1H, 8.85 (1H, d, J=6.9Hz) APT-ES MASS (Negative) 1408 Elemental Analysis Calcd. for C 6 4 H8 6
N
1 2
O
20
S
2 -7H 2
O:
C 50-10, H 6.52, N 10.96 Found: C 50.29, M 6.48, N 10.77 Example 57 IR (KBr) 1666, 1649, 1632, 1554, 1541, 1514, 1450, 1443, 1419, 1240 crrJ 1 NMR (DMSO-d 6 fD 2 0, 0.7-1.3 (17H, in), 1.3-2.6 (7H1, in), 2.7-4.5 (35H, mn), 4.7-4.9 (2H, in), 6.7-6.9 (2H, in), 7.0-7.2 (3H, mn), 7.87 (2H, d, J=8.6Hz), 8.0-8.2 (4H, mn) EST MASS (Positive): 1453.4 (M+2Na) 2 Elemental Analysis Calcd. for C 6 4
H
8 8
N
1 2 0 2 0 2 6H 2 0: C 50.65, H 6.64, N 11-07 Found: C 50.28, H 6.61, N 10.80 Example 58 Major compound: ESI MASS (Negative): 1538.6 (M-1i) Minor compound: IR (KBr) 3352, 1659,1635, 1606, 1529, 1444, 1417, 1274, 1238 cm-1 ESI MASS (Negative): 1338.6 (M-1l) Elemental Analysis Calcd. for C 7
,H
1 0 2
N
1 2
O
2 2
S
2 -7H 2
O:
C 51.19, H 7.02, N 10.09 Found: C 51.19, H 6.95, N 9.73 Example 59 Major compound: EST MASS (in/z) (Positive) 1598.3 (M+2Na) 2 WO 01/60846 PCT/JPOI/01204 177 Minor compound: ESI MASS (in/z) (Negative): 1551.6 (M-2H) 2 Example IR 1664, 1635, 1605, 1446, 1410, 1350, 1329, 1281 c- NMR (DMSO-d 6 D320, 6) 0. 98 (3H, d, J=6. 71z) 1. 10 (3H, di, J=5.9Hz), 1.1-2.6 (21H, mu), 2.8-4.5 (31H, mn), 4.7- 4.9 (2H, mn), 6.7-6.9 (2H, in), 7.0-7.2 (3H, in), 7.85 (2H, di, J=8.9Hz), 8.0-8.2 (4H, m) ESI MASS (in/z) (Negative) 1409.4 (M-1l) Elemental Analysis Caicd. for C 6 4
H
8 7
N
1 1 0 2 1
S
2 -6H 2 0: C 50.62, H 6.57, N 10.15 Found: C 50.40, H 6.61, N 9.92 Example 61 IR (KBr): 2937, 1676, 1651, 1556, 1541, 1514, 1452, 1441, 1419 cin' NMR (DMSO-d 6
+D
2 0, 0.90 (3H, d, J=6.8Hz), 1.10 (3H, d, J=5.7Hz), 1.2-2.6 (17H, in), 2.8-4.5 (37H, mn), 4.7- 4.9 (2H, mn), 6.7-6.9 (2H, in), 7.0-7.2 (3H, mn), 7.85 d, J=8.6Hz), 8.0-8.2 (4H, mn) ESI MASS (Negative): 1427.5 (M-lI) Elemental Analysis Calcd. for C 6 4
H
8 9
N
1 1 0 2 2
S
2 '5.5H 2 0: C 50.32, H 6.60, N 10.09 Found: C 50.31, H 6.72, N 10.04 Example 62 IR 1633, 1606, 1529, 1518, 1444, 1419, 1279, 1252 cm-l NMR (DMSO-d 6
+D
2 0, i) 0.-98 (3H, di, J 6. 7Hz) 1. 10 (3H, d, J=5.8Hz), 1.2-2.6 (19H, mn), 2.8-4.6 (37H, mn), 4.7- 4.9 (2H, in), 6.7-6.9 (2H, mn), 7.0-7.2 (3H, mn), 7.85 (2H, di, J=8.8Hz), 8.0-8.2 (4H, mn) WO 01/60846 PCT/JP01/01204 178 ESI MASS (m/z)(Negative): 1441.5 Elemental Analysis Calcd. for C6 5
H
9 1
N
11 0 2 2
S
2 "7H 2 0: C 49.77, H 6.75, N 9.82 Found: C 49.80, H 6.68, N 9.80 Example 63 IR (KBr): 2935, 1633, 1606, 1529, 1518, 1444, 1419, 1273, 1232 cm 1 NMR (DMSO-d 6
+D
2 0, 0.98 (3H, d, J=6.8Hz), 1.10 (3H, d, J=6.3Hz), 1.2-2.6 (16H, 2.7-4.5 (38H, 4.7- 4.9 (2H, 6.7-6.9 (2H, 7.0-7.2 (3H, 7.85 (2H, d, J=7.8Hz), 8.0-8.2 (4H, m) ESI MASS (m/z)(Negative): 1427.4 Elemental Analysis Calcd. for C 64
H
8 9
N
11 0 2 2
S
2 -6H 2 0: C 50.02, H 6.62, N 10.03 Found: C 49.99, H 6.73, N 9.67 Example 64 IR (KBr): 1659, 1633, 1605, 1547, 1529, 1518, 1444, 1419 cm 1 NMR (DMSO-d 6
+D
2 0, 0.98 (3H, d, J=6.8Hz), 1.11 (3H, d, J=5.7Hz), 1.2-2.6 (18H, 2.8-4.5 (38H, 4.7- 4.9 (2H, 6.7-6.9 (2H, 7.0-7.2 (3H, 7.84 (2H, d, J=8.7Hz), 8.0-8.2 (4H, m) ESI MASS (m/z)(Negative): 1441.5 Elemental Analysis Calcd. for C 65
H
91
N
1 1 0 2 2
S
2 -6H 2 0: C 50.34, H 6.69, N 9.94 Found: C 50.12, H 6.78, N 9.87 Example IR (KBr): 1664, 1628, 1605, 1529, 1444, 1408, 1281, 1252 cm 1 NMR (DMSO-d 6
-+D
2 0, 0.91 (3H, d, J=6.8Hz), 0.98 (3H, d, J=6.8Hz), 1.10 (3H, d, J=5.9Hz), 1.3-2.7 (16H, m), 2.8-4.5 (34H, 4.7-4.9 (2H, 6.7-6.9 (2H, m), 6.9-7.1 (3H, 8.0-8.2 (4H, 8.73 (1H, d, J=2.6Hz) WO 01/60846 PCT/JP01/01204 179 ESI MASS (m/z)(Negative): 1408.5 (M-2H) 2 Elemental Analysis Calcd. for C 6 3
H
87
N
1 3 0 2 0 S2-8H 2 0: C 48.67, H 6.68, N 11.71 Found: C 48.86, H 6.64, N 11.44 Example 66 IR (KBr): 1664, 1635, 1628, 1605, 1444, 1408, 1281, 1259 cm 1 NMR (DMSO-d 6
+D
2 0, 0.86 (3H, d, J=6.3Hz), 0.98 (3H, d, J=6.9Hz), 1.10 (3H, d, J=5.7Hz), 1.1-1.4 (5H, m), 1.6-2.7 (11H, 2.8-4.5 (34H, 4.7-4.9 (2H, m), 6.7-6.9 (2H, 6.9-7.1 (3H, 8.0-8.2 (4H, 8.72 (1H, d, ESI MASS (m/z)(Negative): 1408.6 (M-2H)2- Elemental Analysis Calcd. for C 6 3
H
87
N
1 3 0 20
S
2 -7H 2 0: C 49.24, H 6.62, N 11.85 Found: C 49.05, H 6.73, N 11.48 Example 67 IR (KBr): 3352, 1664, 1635, 1603, 1444, 1408, 1281, 1250 cm- 1 NMR (DMSO-d 6
+D
2 0, 0.85 (3H, t, J=7.4Hz), 0.98 (3H, d, J=6.8Hz), 1.10 (3H, d, J=5.9Hz), 1.3-2.6 (18H, m), 2.8-4.5 (34H, 4.7-4.9 (2H, 6.7-6.9 (2H, m), 6.9-7.1 (3H, 8.0-8.2 (4H, 8.73 (1H, d, J=2.6Hz) ESI MASS (m/z)(Negative): 1423.5 Elemental Analysis Calcd. for C 64
H
8 9 NI30 2 0
S
2 -6H 2 0: C 50.15, H 6.64, N 11.88 Found: C 49.99, H 6.74, N 11.61 Example 68 IR (KBr): 1664, 1628, 1603, 1529, 1444, 1408, 1281, 1248 cm-1 NMR (DMSO-d 6
+D
2 0, 0.7-1.3 (12H, 1.6-2.6 (15H, m), 2.7-4.4 (34H, 4.7-4.9 (2H, 6.6-6.8 (2H, m), WO 01/60846 PCT/JP01/01204 180 6.8-7.0 (3H, 7.9-8.1 (4H, 8.66 (1H, d, ESI MASS (m/z)(Negative): 1423.5 Elemental Analysis Calcd. for C 6 4
H
8 9
N
13 0 2 0
S
2 -6H 2 0: C 50.15, H 6.64, N 11.88 Found: C 49.95, H 6.74, N 11.47 Example 69 IR (KBr): 1658, 1635, 1549, 1529, 1518, 1468, 1446, 1277, 1043 cm 1 NMR (DMSO-d6+D 2 0, 0.97 (3H, d, J=6.8Hz), 1.0-1.4 (9H, 1.5-2.6 (15H, 2.7-4.5 (31H, 4.7-4.9 (2H, 6.7-6.9 (2H, 7.0-7.1 (1H, 7.49 (2H, d, J=8.6Hz), 7.8-8.1 (6H, 8.86 (1H, s) ESI MASS (m/z)(Negative): 1432.4 Elemental Analysis Calcd. for C66H8gN12020S2-6H20 C 51.42, H 6.54, N 10.90 Found: C 51.36, H 6.65, N 10.50 Example IR (KBr): 3493, 3462, 3433, 3350, 1659, 1635, 1613, 1529, 1518, 1466, 1446 cm 1 NMR (DMSO-d 6
+D
2 0, 0.90 (3H, d, J=6.7Hz), 0.98 (3H, d, J=6.6Hz), 1.11 (3H, d, J=5.3Hz), 1.3-2.7 (16H, m), 2.8-4.5 (34H, 4.7-4.9 (2H, 6.7-6.9 (2H, m), 7.0-7.2 (3H, 7.78 (2H, d, J=8.7Hz), 7.9-8.1 (4H, 8.78 (1H, s) ESI MASS (m/z)(Negative): 1447.5 Elemental Analysis Calcd. for C 6 6
H
8 9
N
1 3 0 20
S
2 8H 2 0: C 49.77, H 6.64, N 11.43 Found: C 50.09, H 6.68, N 11.14 Example 71 NMR (DMSO-d 6 0.8-2.8 (40H, 2.8-4.6 (28H, m), 4.7-5.4 (9H, 6.6-6.85 (2H, 6.9-7.1 (3H, m), 7.3-8.5 (12H, 8.6-8.8 (2H, m) WO 01/60846 PCT/JP01/01204 181 MASS 1391 Elemental Analysis Calcd. for C 6 7
H
9 6
N
1 0 0 2 0 S7H 2 0: C 52.95, H 7.30, N 9.22 Found: C 52.88, H 7.33, N 9.22 The following compound was obtained according to a similar manner to that of Example 54.
Example 72 NMR (DMSO-d 6
+D
2 0, 0.7-4.5 (67H, 4.65-4.85 (2H, m), 6.3-6.45 (1H, 6.5-6.7 (2H, 7.12 (2H, d, J=8.8Hz), 7.6-7.8 (4H, 7.95 (2H, d, J=8.4Hz) ESI MASS 1311 Elemental Analysis Calcd. for C 67
H
96
N
1 0 0 17 -3HCl-10H 2 0: C 50.20, H 7.48, N 8.74 Found: C 50.28, H 7.15, N 8.67 The following compounds [Examples 73 to 87] were obtained according to a similar manner to that of Example 33.
Example 73 NMR (DMSO-d 6 0.84 (9H, 0.97 (3H, d, 1.0-1.4 (8H, 1.6-2.8 (18H, 2.8-4.6 (28H, m), 4.7-5.4 (9H, 6.6-6.8 (2H, 6.9-7.1 (3H, m), 7.3-8.5 (12H, 8.6-8.8 (2H, m) MASS 1365 Elemental Analysis Calcd. for C 65
H
9 4
N
1 00 2 0 S-7H 2 0: C 52.27, H 7.29, N 9.38 Found: C 52.15, H 7.30, N 9.30 Example 74 IR (KBr): 1649, 1539, 1514, 1454, 1439, 1045 cm 1 NMR (DMSO-d 6 0.7-1.4 (16H, 1.6-2.8 (18H, m), 2.8-4.6 (28H, 4.7-5.5 (9H, 6.6-6.8 (2H, m), 6.9-7.1 (3H, 7.3-8.5 (12H, 8.5-8.8 (2H, m) WO 01/60846 PCT/JPOI/01204 182 MASS 1337 Elemental Analysis Calcd. for C 6 3 Hg 0
N
1 0 0 2 0 5-9H 2 0: C 50.39, H 7.25, N 9.33 Found: C 50.64, H 6.96, N 9.24 Example IR (KBr): 1666, 1649, 1632, 1539, 1514, 1454, 1238 cm'1 NMR (DMSO-d 6 0.85 (3H, t, J=7.0Hz), 0.97 (3H, d, J=6.6Hz), 1.09 (3H, d, Jrr5.5Hz), 1.2-2.75 (25H, mn), 2.8-4.6 (28H, in), 4.7-5.4 (9H, in), 6.6-6.8 (2H, in), 6.9-7.1 (3H, in), 7.3-8.5 (12H, mn), 8.6-8.8 (2H, in) MASS 1337 Elemental Analysis Calcd. for C 6 3
H
9 0
N
1 0
O
2 0 S-7H 2
O:
C 51.63, H 7.15, N 9.56 Found: C 51.74, H 7.07, N 9.52 Examp le 76 TR (KBr) 1666, 1649, 1632, 1539, 1514, 1236 cmJ 1 NMR (DMSO-d 6 0.98 (3H, d, J=6o.6Hz), 1.10 (3H, d, J=5.7H4z), 2.8-4.6 (64H-, in), 4.7-5-4 (9H, in), 6.6-6.85 (2H, in), 6.9-7.15 (3H, in), 7.3-8.5 (12H, mn), 8.6-8.8 (2 H, in) MASS 1421 Elemental Analysis Calcd. for C 6 8
H
9
BN
1 0
O
2 1 S-8H 2
O:
C 52.10, H 7.33, N 8.93 Found: C 52.18, H 7.22, N 8.85 Example 77 Examp~le 78 IR (KBr) 1666, 1632, 1539, 1514, 1452, 1236 c-- NMR (DMSQ-d 6 0.7-2.7 (38H, in), 2.8-4.6 (32H, in), 4.7-5.4 in), 6.6-6.85 (3H, in), 6.9-7.1 (2H, mn), 7.3-8.5 (12H, in), 8.6-8.8 (2H, in) MASS 1421 WO 01/60846 PCT/JP01/01204 183 Elemental Analysis Calcd. for C 6 8
H
98
N
1 0 0 2 1 S-8H 2 0: C 52.10, H 7.33, N 8.94 Found: C 52.10, H 7.17, N 9.33 Example 79 IR (KBr): 1632, 1539, 1516, 1452, 1238 cm 1 NMR (DMSO-d 6 0.8-1.4 (16H, 1.6-2.8 (22H, m), 2.8-5.55 (32H, 4.7-5.4 (9H, 6.65-6.85 (2H, m), 6.9-7.1 (3H, 7.3-8.5 (12H, 8.5-8.8 (2H, m) MASS 1421 Elemental Analysis Calcd. for C 6 8
H
98
N
1 0 0 2 1 S.8H 2 0: C 52.10, H 7.33, N 8.94 Found: C 51.82, H 7.17, N 9.23 Example IR (KBr): 1666, 1645, 1632, 1539, 1514, 1452, 1240 cm 1 NMR (DMSO-d 6 0.98 (3H, d, J=6.8Hz), 1.10 (3H, d, 1.4-2.75 (23H, 2.8-4.5 (31H, 4.7-5.4 (9H, 6.65-6.9 (4H, 6.9-7.1 (3H, 7.15 (2H, d, J=8.7Hz), 7.3-8.5 (12H, 8.6-8.8 (2H, m) MASS 1415 Elemental Analysis Calcd. for C 68
H
92
N
1 00 2 1 S-12H 2 0: C 49.99, H 7.16, N 8.57 Found: C 49.86, H 6.81, N 8.96 Example 81 IR (KBr): 1632, 1539, 1514, 1452, 1275 cm-1 NMR (DMSO-d 6 0.97 (3H, d, J=6.7Hz), 1.10 (3H, d, J=5.7Hz), 1.2-2.8 (23H, 2.8-4.55 (31H, 4.7-5.4 (9H, 6.6-6.9 (4H, 6.9-7.1 (3H, 7.14 (2H, d, J=8.7Hz), 7.3-8.5 (12H, m) 8.6-8.8 (2H, m) MASS 1415 Elemental Analysis Calcd. for C 6 8
H
92
N
10 0 2 1
S-H
2 0: C 52.30, H 6.97, N 8.97 Found: C 52.48, H 6.79, N 9.44 WO 01/60846 PCT/JP01/01204 184 Example 82 IR (KBr): 1676, 1649, 1632, 1539, 1514, 1456, 1236 cm- 1 NMR (DMSO-d 6 0.88 (6H, 0.97 (3H, d, J=6.6Hz), 1.10 (3H, d, J=5.4Hz), 1.1-2.8 (22H, 2.8-4.6 (28H, m), 4.7-5.5 (9H, 6.6-6.8 (2H, 6.9-7.1 (3H, m), 7.3-8.8 (14H, m) MASS 1339 Elemental Analysis Calcd. for C 63
H
9 0
N
1 0 0 2 0 SS8H 2 0: C 51.00, H 7.20, N 9.44 Found: C 51.31, H 7.16, N 9.44 Example 83 IR (KBr): 1664, 1635, 1626, 1605, 1446, 1408, 1350, 1329 cm 1 NMR (DMSO-d 6
+D
2 0, 0.98 (3H, d, J=6.8Hz), 1.09 (3H, d, J=5.7Hz), 1.2-2.8 (24H, 2.8-4.5 (37H, 4.7- 4.9 (2H, 6.7-6.9 (2H, 7.0-7.1 (3H, 7.62 (2H, d, J=8.7Hz), 7.70 (2H, d, J=8.4Hz), 7.93 (2H, d, J=8.4Hz) ESI MASS (m/z)(Negative): 1407.6 (M-2H) 2 Elemental Analysis Calcd. for C 6 7
H
9 6
N
1 0 0 21 S-6H 2 0: C 53.02, H 7.17, N 9.23 Found: C 52.98, H 7.28, N 9.13 Example 84 IR (KBr): 1664, 1628, 1606, 1531, 1497, 1446, 1281, 1238 cm- 1 NMR (DMSO-d 6
+D
2 0, 0.97 (3H, d, J=6.7Hz), 1.10 (3H, d, J=5.4Hz), 1.3-2.7 (24H, 2.8-4.5 (37H, 4.7- 4.9 (2H, 6.7-6.9 (2H, 7.0-7.2 (3H, 7.62 (2H, d, J-8.6Hz), 7.70 (2H, d, J-8.5Hz), 7.93 (2H, d, J= ESI MASS (m/z)(Negative): 1408.4 (M-l) Elemental Analysis Calcd. for C 67
H
9 6
N
1 0 0 2 1 S-5H 2 0: WO 01/60846 PCT/JP01/01204 185 C 53.66, H 7.12, N 9.34 Found: C 53.58, H 7.34, N 9.15 Example IR (KBr): 1664, 1628, 1606, 1529, 1497, 1446, 1408, 1281, 1238 cm 1 NMR (DMSO-d 6
+D
2 0, 0.97 (3H, d, J=6.9Hz), 1.10 (3H, d, 1.5-4.5 (52H, 4.7-4.9 (2H, 6.7-6.9 (2H, 6.9-7.1 (3H, 7.2-7.5 (5H, 7.62 (2H, d, J=8.6Hz), 7.71 (2H, d, J=8.6Hz), 7.93 (2H, d, J=8.3Hz) ESI MASS (m/z)(Negative): 1415.4 (M-2H) 2 Elemental Analysis Calcd. for C 6 8
H
92
N
1 0 0 2 1 S-6H 2 0: C 53.53, H 6.87, N 9.18 Found: C 53.55, H 6.91, N 9.00 Example 86 NMR (DMSO-d 6 0.9 (3H, d, J=6.BHz), 0.98 (3H, d, J=6.8Hz), 1.10 (3H, d, J=6.1Hz), 1.3-2.7 (24H, 2.8-4.6 (29H, 4.7-5.3 (9H, 6.6-6.8 (2H, 6.9-7.2 (3H, m), 7.3-8.2 (11H, 8.4-8.6 (1H, 8.7 (1H, s), 8.8-8.95 (1H, m) MASS 1421 Elemental Analysis Calcd. for C 65
H
9 0 N1 2 0 20
S
2 8' 2 0: C 49.80, H 6.81, N 10.72 Found: C 50.07, H 6.74, N 10.73 Example 87 IR (KBr): 3351.7, 2931.3, 2854.1, 1658.5, 1635.3, 1546.6, 1531.2, 1496.5 cm-1 NMR (DMSO-d6+D 2 0, 0.97 (3H, d, J=7Hz), 0.8-4.5 complex 3.01 (3H, 4.79-4.81 (2H, 6.72 (1H, d, J=8Hz), 6.75-6.80 (1H, 7.01 (1H, 7.03 (2H, d, J=8Hz), 7.61 (2H, d, J=8Hz), 7.69 (2H, d, J=8.4Hz), 7.94 (2H, d, J=8.4Hz) WO 01/60846 PCT/JP01/01204 186 ESI MASS (Negative): 1435.7 Example 88 To a solution of a mixture of starting compound (88) 1,3-dihydroxyacetone (1.19 g) and acetic acid (1.14 ml) in a mixture of methanol (120 ml) and DMF (55 ml) was added sodium cyanoborohydride (835 mg) with stirring at ambient temperature, and the mixture was stirred at the same temperature overnight.
To a reaction mixture was poured into ethyl acetate (700 ml).
The resulting precipitates were collected by filtration, washed with ethyl acetate (100 ml) and dried in vacuo. The precipitates were dissolved in a mixture of 30% aqueous acetonitrile (800 ml) and 1N sodium hydroxide (5 ml). The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (440 ml) eluting in turn with water and aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (88) (5.22 g).
IR (KBr): 1632, 1535, 1518, 1443, 1269, 1082, 1047 cm 1 NMR (DMSO-d 6
+D
2 0, 0.82 (3H, d, J=6.83Hz), 0.97 (3H, d, J=6.81Hz), 1.02 (3H, d, J=6.18Hz), 1.24 (26H, s), 1.35-2.45 (14H, 2.75-3.40 (5H, 3.60-4.50 4.70-4.90 (2H, 6.65-6.80 (2H, 7.01 (1H, s) ESI MASS (m/z)(Positive): 1088.4 (M++Na) Example 89 To a solution of starting compound (89) (4.0 g) in DMF ml) were successively added diisopropylethylamine (1.45 ml) and 9-fluorenylmethyl chloroformate (1.03 and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was poured into water (200 ml). The solution was purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting in turn WO 01/60846 PCT/JP01/01204 187 with a mixture of saturated aqueous sodium chloride (400 ml), saturated aqueous sodium hydrogen carbonate (50 ml) and water (400 ml), and aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (89) (2.82 g).
IR (KBr): 1666, 1632, 1518, 1446, 1273, 1246, 1082, 1047 cm 1 NMR (DMSO-d 6
+D
2 0, 0.80-1.10 (9H, 1.23 (26H, s), 1.35-2.45 (12H, 2.60-3.40 (6H, 3.60-4.55 (18H, 4.65-4.90 (2H, 6.65-6.85 (2H, 6.97 (1H, 7.30-7.50 (4H, 7.60-7.95 (4H, m) ESI MASS (m/z)(Negative): 1423.7 (M+-Na) Elemental Analysis Calcd. for C 69
H
99
N
6 0 22 SNa-6H 2 0: C 53.27, H 7.19, N 7.20 Found: C 53.45, H 7.21, N 7.10 Example To a solution of starting compound (90) (1.21 g) in DMF ml) were successively added diisopropylethylamine (0.26 ml) and di-tert-butyl dicarbonate (285 mg) and the mixture was stirred at ambient temperature overnight. The reaction mixture was poured into a mixture of pH 6.86 standard buffer solution (150 ml), saturated aqueous sodium chloride (50 ml) and saturated aqueous sodium hydrogen carbonate (20 ml). The mixture was purified by ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (200 ml) column chromatography, eluting with aqueous acetonitrile The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (90) (1.19 g).
IR (KBr): 1662, 1632, 1535, 1518, 1444, 1367, 1272, 1250 cm 1 NMR (DMSO-d 6
+D
2 0, 0.85 (3H, d, J=6.76Hz), 0.96 (3H, d, J=6.77Hz), 1.04 (3H, d, J=5.50Hz), 1.23 (26H, 1.37 WO 01/60846 PCT/JP01/01204 188 (9H, 1.40-1.50 (2H, 1.55-2.50 (10H, m), 2.80-3.40 (6H, 3.50-4.45 (14H, 6.65-6.80 (2H, 6.96 (1H, s) ESI MASS (m/z)(Negative): 1301.6 (M+-Na) Example 91 To a solution of a mixture of starting compound (91) 2-phenyl-l,3-dioxane-5-carbaldehyde (0.52 g) and acetic acid (0.35 ml) in a mixture of methanol (30 ml) and DMF (14 ml) was added sodium cyanoborohydride (254 mg) with stirring at ambient temperature and the mixture was stirred at the same temperature for 6 hours. The reaction mixture was poured into ethyl acetate (300 ml). The resulting precipitates were collected by filtration, washed with ethyl acetate (50 ml) and dried in vacuo. The precipitates were dissolved with pH 6.86 standard buffer solution (100 ml) and acetonitrile (200 ml) and the solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (220 ml) eluting in turn with water (1 acetonitrile in water and 30% acetonitrile in water. The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (91) (2.27 g).
NMR (DMSO-d 6
+D
2 0, 6) 0.96 (3H, d, J=6.56Hz), 1.07 (3H, d, J=5.42Hz), 1.33, 1.37 (9H, broad 1.50-2.05 (6H, 2.10-2.45 (2H, 2.60-3.50 (6H, 3.75-4.50 (16H, m) 4.75-4.85 (2H, m) 5.44, 5.55 (1H, broad s), 6.75 (2H, 7.38 (5H, br s) ESI MASS (m/z)(Negative): 1189.3 (M++Na) Example 92 A solution of starting compound (92) (2.26 10% palladium on carbon (50% including water) (2.0 g) and 10% palladium hydroxide on carbon (2.0 g) in a mixture of methanol (45 ml) and water (23 ml) was hydrogenated under an atmospheric pressure of WO 01/60846 PCT/JP01/01204 189 hydrogen with stirring at ambient temperature for 6 hours. The catalyst was filtered off and washed with a mixture of methanol and water (1:1 v/v) (50 ml), and the filtrates were combined.
The mixture was evaporated in vacuo and dissolved in water (200 ml). The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (220 ml) eluting with water and 30% acetonitrile in water. The first fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give the object compound (92) (1.84 g).
NMR (DMSO-d 6
+D
2 0, 0.97 (3H, d, J=6.75Hz), 1.07 (3H, d, J=5.76Hz), 1.35 (9H, 1.45-2.00 (6H, 2.10-2.45 (3H, 2.70-3.45 (9H, 3.55-4.55 (17H, m), 4.75-4.85 (2H, 6.65-6.80 (2H, 7.02 (1H, s) ESI MASS (Positive): 1123.3 1101.3 (M++2Na), Example 93 To a solution of a mixture of starting compound (93) (1.83 g) and diisopropylethylamine (0.65 ml) in DMF (20 ml) was added 9-fluorenylmethyloxycarbonyl chloride (483 mg) with stirring at ambient temperature and the mixture was stirred at the same temperature for 2 hours. The reaction mixture was poured into water (300 ml). The mixture was adjusted to pH 7.5 with IN HC1 and washed with ethyl acetate (100 ml). The aqueous layer was evaporated to remove organic solvent. To a concentrated solution were added saturated aqueous sodium hydrogen carbonate (50 ml) and 5% aqueous sodium chloride (20 ml). The solution was subjected to column chromatography on ODS (Daiso-gel, SP- 120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (220 ml) eluting in turn with water (1 20% acetonitrile in water (1 L) and 30% acetonitrile in water (1 The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (93) (2.106 g) WO 01/60846 PCT/JP01/01204 190 NMR (DMSO-d6+D 2 0, 0.89 (3H, br 1.07 (3H, br 1.34 (9H, 1.45-2.50 (10H, 2.60-3.40 (13H, m), 3.70-4.50 (14H, 4.65-4.90 (2H, 6.65-6.80 (2H, 6.99 (1H, 6.95-7.48 (4H, 7.60-7.70 (2H, 7.85-7.95 (2H, m) ESI MASS (m/z)(Positive): 1345.3 (M +Na) Example 94 To a solution of a mixture of starting compound (94) (2.10 g) and triethylsilane (2.03 ml) in dichloromethane (35 ml) was dropwise added trifluoroacetic acid (3.70 ml) with stirring under ice-cooling and the mixture was stirred at ambient temperature for 1 hour. The reaction mixture was poured into a mixture of pH 6.86 standard buffer solution (150 ml) and saturated aqueous sodium hydrogen carbonate (20 ml) The mixture was adjusted to pH 8 with saturated aqueous sodium carbonate.
The organic layer was separated and concentrated in vacuo to remove organic solvent. The solution was subjected to column chromatography on ODS (Daiso-gel, SP-120-40/60-ODS-B (Trademark: prepared by Daiso Co., Ltd.)) (220 ml) eluting in turn with water (1 10% acetonitrile in water (800 ml), acetonitrile in water (1 L) and then 30% acetonitrile in water (1 The fractions containing the object compound were collected and evaporated under reduced pressure to remove acetonitrile. The residue was lyophilized to give object compound (94) (1.704 g).
IR (KBr): 1668, 1633, 1539, 1516, 1440, 1273, 1082, 1045 cm 1 NMR (DMSO-d 6
+D
2 0, 0.89 (3H, br 1.05-1.20 (3H, m), 1.30-2.40 (8H, 2.60-3.40 (10H,m), 3.50-4.45 (16H, 4.60-4.85 (2H, 6.73 (2H, br 6.97 (1H, s), 7.25-7.48 (4H, 7.66 (2H, d, J=7.12Hz), 7.88 (2H, d, J=7.24Hz) ESI MASS (m/z)(Positive): 1199.4 1200.4 (M Elemental Analysis Calcd. for C 5 4
H
8 4
N
8 0 2 7 S-6H 2 0: P:\OPER\JgcD2001234095 claims doc.23/07/04 -191- C 49.53, H 6.47, N 8.56 Found: C 49.30, H 6.26, N 8.49 The following compound was obtained according to a similar manner to that of Example 33.
Example IR (KBr): 1664, 1628, 1605, 1446, 1417, 1279, 1084, 1047 cm-1 NMR (DMSO-d 6
+D
2 0, 0.8-1.3 (12H, 1.5-2.6 (16H, m), 2.8-4.5 (32H, 4.7-4.9 (2H, 6.7-6.9 (2H, m), 7.0-7.2 (3H, 7.85 (2H, d, J=8.6Hz), 8.0-8.2 (4H, m) ESI MASS (m/z)(Negative): 1423.5 (M-l) Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.
The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form or suggestion that that prior art forms part of the common general knowledge in Australia.

Claims (7)

1. A polypeptide compound of the following general formula OH NH H 3 C NH-R 1 \N 0== NH 0 CH 3 0- CN R 4 NH OH S OH R s 5 R6 (I) wherein R 1 is benzoyl substituted with a suitable substituent selected from the group consisting of thiadiazolyl substituted with phenyl having phenyl substituted with morphlino having CI-C6 alkyl, thiadiazolyl substituted with phenyl having a suitable substituent selected from the group consisting of Ci-C 6 alkoxy CI-C6 alkoxy and Ci-C6 alkoxy C7-C 20 alkoxy, piperazinyl substituted with phenyl having piperidyl substituted with a suitable substituent selected from the group consisting of phenyl having Ci-C 6 alkoxy Ci-C 6 alkoxy, cyclo C3-C6 alkyloxy and CI-C6 alkoxy C 1 -C 6 alkylthio, piperazinyl substituted with phenyl having phenyl substituted with morpholino having Ci-C6 alkyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with a suitable substituent selected P:\OPERUgc\2001234095 claims doc21/07/04 -193- from the group consisting of Ci-C6 alkoxy Ci-C6 alkoxy and C1-C6 alkoxy Ci-C6 alkylthio, imidazothiadiazolyl substituted with phenyl having Ci-C6 alkoxy Ci-C6 alkoxy, phenyl substituted with piperazinyl having phenyl substituted with morpholino having CI-C6 alkyl, isoxazolyl substituted with phenyl having Ci-C6 alkoxy C1-C6 alkoxy, isoxazolyl substituted with phenyl having C17-C20 alkoxy substituted with morpholino having Ci-C6 alkyl, thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo C3-C6 alkyl which has one or more suitable substituent(s) selected from the group consisting of Ci-C6 alkyl, C2-C6 alkenyl, Ci-C6 alkoxy C17-C20 alkoxy and phenyl, thiadoazolyl substituted with phenyl having piperazinyl substituted with Ci-C6 alkyl having cyclo C3-C6 alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with one or more suitable substituent(s) selected from the group consisting of cyclo C3-C6 alkyloxy, Ci-C6 alkoxy Ci-C6 alkoxy and C1-C6 alkoxy Ci-CE alkoxy Ci-C6 alkyl, thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo C3-C6 alkyl and Ci-C6 alkoxy, thiadiazolyl substituted with pyridyl having piperazinyl substituted with cyclo C3-C6 alkyl having CI-C6 alkyl, imidazothiadiazolyl substituted with phenyl having piperidyl substituted with cyclo C3-C6 alkyl, imidazothiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo C3-C6 alkyl having Ci-C6 alkyl, and P:\OPERUgc\2001234095 claimsdoc-2307/04 -194- phenyl substituted with piperazinyl having cyclo C3-C6 alkyl substituted with one or more suitable substitutent(s) selected from the group consisting of cyclo C3-C6 alkyl which may have C1-C6 alkoxy, Ci-C6 alkyl, Ci-C6 alkoxy and phenyl which may have Cl-C 6 alkoxy, R 2 is hydrogen, R 3 is C1-C6 alkyl which has two hydroxy, R 4 is hydrogen or hydroxy; R 5 is hydroxy or hydroxysulfonyloxy; and R 6 is hydroxy, or a salt thereof.
2. A compound of the claim 1, wherein R 1 is benzoyl which has thiadiazolyl substituted with phenyl having piperazinyl substituted with cyclo C3-C6 alkyl which has C1-C6 alkyl, benzoyl which has thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo C3-C6 alkyloxy, benzoyl which has phenyl substituted with piperazinyl having cyclo C3-C6 alkyl substituted with cyclo C3-C6 alkyl and Ci-C6 alkoxy, or benzoyl which has thiadiazolyl substituted with phenyl having piperidyl substituted with cyclo C3-C6 alkyl, R 2 is hydrogen, R 3 is Ci-C6 alkyl which has two hydroxy, R 4 is hydrogen or hydroxy; R 5 is hydroxy or hydroxysulfonyloxy; and R 6 is hydroxy.
3. A compound of the claim 1, wherein R 1 is benzoyl which has thiadiazolyl substituted with phenyl having a suitable substituent selected from the group consisting of P:IOPERUgU\00I 234095 cIims.doc.2307/04 -195- piperazinyl substituted with cyclo (C 3 -C 6 )alkyl which has (Ci-C) alkyl, and piperidyl substituted with cyclo (C 3 -C 6 )alkyl and (C 1 -C 6 )alkoxy, R 2 is hydrogen, R 3 is (C 1 -C 6 )alkyl which has two hydroxy, R 4 is hydrogen, R 5 is hydroxysulfonyloxy, and R 6 is hydroxy.
4. A compound of the claim 3, wherein R 1 is 4-[5-[4-[4-(4-methylcyclohexyl)-piperazin-1-yl)phenyl]- 1,3,4-thiadiazol-2-yl]benzoyl, R 2 is hydrogen, R 3 is 2-hydroxy-1-(hydroxymethyl)ethyl, R 4 is hydrogen, R 5 is hydroxysulfonyloxy, and R 6 is hydroxy.
5. A compound of the claim 3, wherein R 1 is 4-[5-[4-(4-methoxy-4-cyclohexyl-piperidin-1-yl)phenyl]- 1,3,4-thiadiazol-2-yl]benzoyl, R 2 is hydrogen, R 3 is 2-hydroxy-1-(hydroxymethyl)ethyl, R 4 is hydrogen, R 5 is hydroxysulfonyloxy, and R 6 is hydroxy.
6. A process for preparing a polypeptide compound of claim 1, or a salt thereof, which comprises, reacting a compound (II) of the formula: P:\OPERUgc\20123409S claim.do-26/07/04 -196- Re' (II) wherein R 1 R 4 R 5 and R 6 are defined in claim 1, or its reactive derivative at the amino group or a salt thereof, with a compound (III) of the formula: R 3 =0 (III) wherein R 3 is defined in claim 1, or its reactive derivative or a salt thereof, to give a compound (Ia) of the formula: OH O NH I NNH NH OH N OH thereof, or thereof, or P:\OPER~gv\2001234095 claimsdoc26/07/04 197 (ii) subjecting a compound (Ib) of the formula: OH OH 0 NH H 3 NH-R 1 N 0 HO 0 HN OH /NJ NH 0 CH 3 R 4 0NH O 0 OH R 6 (Ib) wherein R, R R, R 5 and R 6 are defined in claim 1, R2is acyl group, or a salt thereof, to elimination reaction of the acyl group, to give a compound (Ta) of the formula: (ITa) P\OPERUgc\U001234095 dams.dmc.26/07/4
198- wherein R R 3 R 4 R 5 and R 6 are defined above, or a salt thereof, or (iii) reacting a compound (Ic) of the formula: Re (Ic) wherein R 2 R 3 R 4 R 5 and R 6 are defined in claim 1, or its reactive derivative at the amino group or a salt thereof, with a compound of the formula: R 1 -OH (V) wherein R 1 is defined in claim 1 or its reactive derivative at the carboxy group or a salt thereof, to give a compound (Id) of the formula: P:\OPERUgcU\00 234095 claims doc.2607/04 -199- NH H 3 C NH-R' 2 HO 0 HN OH R3 NH 0 CH 3 0 N R NH OH R 6 (Id) wherein R, R 2 R 3 R, R 5 and R 6 are defined in claim 1, or a salt thereof. 7. A pharmaceutical composition which comprises, as an active ingredient, a compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with pharmaceutically acceptable carriers or excipients. 8. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament. 9. A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as a medicament. A method for the prophylactic and/or therapeutic treatment of infectious diseases caused by pathogenic microorganisms, which comprises administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to a human being or an animal. PAOPERUg\2001234095 claimsdOc-2607104 -200- 11. A commercial package comprising the pharmaceutical composition of claim 7 and a written matter associated therewith, wherein the written matter states that the pharmaceutical composition can or should be used for preventing or treating infectious disease. 12. An article of manufacture, comprising packaging material and the compound identified in claim 1 contained within said packaging material, wherein the said compound is therapeutically effective for preventing or treating infectious diseases, and wherein said packaging material comprises a label or a written material which indicates that said compound can or should be used for preventing or treating infectious diseases. 13. Use of a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in the treatment of infectious diseases caused by pathogenic microorganisms. 14. A compound of claim 1 or a pharmaceutically acceptable salt thereof substantially as hereinbefore described with reference to the Examples. DATED this 2 6 th day of July, 2004 Fujisawa Pharmaceutical Co., Ltd. by DAVIES COLLISON CAVE Patent Attorneys for the Applicant
AU2001234095A 2000-02-21 2001-02-20 Cyclic hexapeptide derivatives Ceased AU2001234095B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001234095A AU2001234095B2 (en) 2000-02-21 2001-02-20 Cyclic hexapeptide derivatives

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
AUPQ5752A AUPQ575200A0 (en) 2000-02-21 2000-02-21 New compound
AUPQ5752 2000-02-21
AUPQ9552 2000-08-21
AUPQ9552A AUPQ955200A0 (en) 2000-08-21 2000-08-21 New compound
AUPR2344 2000-12-28
AUPR2344A AUPR234400A0 (en) 2000-12-28 2000-12-28 New compound
PCT/JP2001/001204 WO2001060846A1 (en) 2000-02-21 2001-02-20 Cyclic hexapeptide derivatives
AU2001234095A AU2001234095B2 (en) 2000-02-21 2001-02-20 Cyclic hexapeptide derivatives

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AU2001234095A1 AU2001234095A1 (en) 2001-11-08
AU2001234095B2 true AU2001234095B2 (en) 2004-09-02

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