AU764109B2 - Novel antibacterial compounds - Google Patents

Novel antibacterial compounds Download PDF

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Publication number
AU764109B2
AU764109B2 AU48885/02A AU4888502A AU764109B2 AU 764109 B2 AU764109 B2 AU 764109B2 AU 48885/02 A AU48885/02 A AU 48885/02A AU 4888502 A AU4888502 A AU 4888502A AU 764109 B2 AU764109 B2 AU 764109B2
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group
compound
doc
methyl
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AU4888502A (en
Inventor
Masatoshi Arai
Hitoshi Hotoda
Masatoshi Inukai
Masakatsu Kaneko
Masaaki Kizuka
Shunichi Miyakoshi
Yasumasa Ogawa
Toshio Takatsu
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Sequella Inc
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Sankyo Co Ltd
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Description

AUSTRALIA
PATENTS ACT 1990 DIVISIONAL APPLICATION NAME OF APPLICANT: Sankyo Company, Limited ADDRESS FOR SERVICE: DAVIES COLLISON CAVE Patent Attorneys 1 Little Collins Street Melbourne, 3000.
INVENTION TITLE: "Novel antibacterial compounds" The following statement is a full description of this invention, including the best method of performing it known to us: New antimicrobial compounds [Technical field] The present invention relates to a compound of formula (XII). (XIII).
(XIV). (XV) or (XVI) and a derivative of a compound of formula (Ia) which have excellent antibiotic activity or a pharmaceutically acceptable salt thereof.
The present invention is also a pharmaceutical composition comprising a compound described above as an active ingredient effective to treat or prevent infectious diseases.
The present invention includes a use of a compound described above in order to prepare a medicament effective to treat or prevent infectious diseases.
The present invention is concerned with a method effective to treat or prevent infectious diseases in warm-blooded animals comprising administering a pharmacologically effective amount of a compound described above to them.
The present invention includes a microorganism capable of producing a compound of formula (XII). (XIV). (XV) or (XVI).
The present invention also includes a process for preparing a compound of formula (XII). (XIV). (XV) or (XVI) using the said microorganism.
[Background of the invention] A P-lactam antibiotic, an amino-glycoside. isoniazid or rifampicin has been conventionally used in treatment or prophylaxis of microbial infections including tubercule bacillus. Recently there have been a lot of bacteria resistant to these antibiotics. It is desirable to develop new compounds which are different type antimicrobial agents from conventional ones.
On the other hand it has been known that capuramycin having a formula shown below exhibits anti-tubercule bacillus activity Antibiotics. 29. 1047-1053 (1986)).
Doc FP9907sl.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 1-32)/0812 00
OH
S ,\OH O H OHCONH 2 HN N 0 N NH HN N o o"rr" H'7 0 0
CH
3 bOH capuramycin We found new compounds of formula (XII). (XIV). (XV) or (XVI).
which do not show any cross resistance to conventional medicaments. in the cultivation products of a microorganism. We prepared the derivatives of compounds described above and capuramycin. We studied the physiological activity of these derivatives for several years and found that these derivatives exhibit excellent antibiotic activity.
The compounds of the present invention can provide a method effective to treat and prevent infection diseases including ones arising from bacteria resistant to the conventional antibiotics. Compounds of formula (XII). (XIV). (XV) or (XVI) are also useful starting materials for preparation of the compounds of the present invention having excellent antibiotic activity.
[Disclosure of the invention] The present invention includes a compound of formula (I)
OH
R0 SH
ONH
HN N 0 N NH X 2RO OH (wherein R' is a methyl group. R 2 is a methyl group. R 4 is a hydroxy group. and X is a methylene group; R' is a methyl group, R 2 is a hydrogen atom. R 4 is a hydroxy group. and X is a methylene group; R' is a methyl group. R 2 is a methyl group, R 4 is a hydrogen atom. and X is a methylene group; Doc FP9907sl.doc P81485/FP-9907(PCTu/tsa-gad-sh/English translation of spec (pages 1-32)/08.12.00 R' is a hydrogen atom, R 2 is a hydrogen atom. R 4 is a hydroxy group, and X is a methylene group; or R' is a methyl group. R 2 is a methyl group. R 4 is a hydroxy group. and X is a sulfur atom) or a pharmaceutically acceptable salt thereof: or a pharmaceutically acceptable ester. ether or N-alkylcarbamoyl derivative of a compound of formula (Ia)
OR
5
R
4 0 2 O H CONH2 Y HN N 0 N NH (Ia) R N 0 0 R OO OR3 X R 2 aO OR 3 (wherein R is a hydrogen atom or a methyl group. Ra is a hydrogen atom. a protecting group for a hydroxy group. or a methyl group, R' is a hvdrogen atom or a protecting group for a hydroxy group. R 4 a is a hydrogen atom. a hydroxy group or a protected hydroxy group, R" is a hydrogen atom or a protecting group for a hydroxy group. and X is a methylene group or a sulfur atom.
with the proviso that when X is a sulfur atom.
R is a methyl group. R 2 is a methyl group. and R 4 is a hydroxy group or a protected hydroxy group: when X is a methylene group. R' is a methyl group. and R 2 a is a hydrogen atom.
R
4 a is a hydroxy group or a protected hydroxy group: or when X is a methylene group and R' is a hydrogen atom.
R
2 a is a methyl group and R 4 a is a hydroxy group or protected hydroxy group): or a phamaceutically acceptable salt thereof.
The present invention is also a pharmaceutical composition comprising a compound described above as an active ingredient effective to treat or prevent infectious diseases.
The present invention includes the use of a compound described above in order to prepare a medicament effective to treat or prevent infectious diseases.
Doc. FP9907sl doc P81485/FP-9907(PCT)/tsa-ead-sh/English translation of spec (pages 1-32)/08.12.00 The present invention is concerned with a method effective to treat or prevent infectious diseases in warm-blooded animals comprising administering a pharmacologically effective amount of a compound described above to them.
The present invention includes a microorganism capable of producing a compound of formula The present invention also includes a process for preparing a compound of formula (I) using the said microorganism.
In the above formulae, the protecting group of "protecting group for a hydroxy group" and "protected hydroxy group" of R 2 a and the like can be removed by a chemical procedure such as hydrogenolysis, hydrolysis, electrolysis or photolysis (hereinafter referred to as a general protecting group) or can be removed by biological method such as hydrolysis in vivo (with the proviso that it is not an ester residue group such as an acyl group). "The protecting group which can be removed by biological method such as hydrolysis in vivo" can be cleaved by biologically method such as hydrolysis in the human body to give a corresponding free acid or a salt thereof. Whether a compound has a protecting group removed in vivo is determined by detection of a corresponding parent compound or a pharmaceutically acceptable salt thereof in the body fluid of a rat or mouse to which it is administered by intravenous injection.
A general protecting group is selected from the group consisting of: S: "tetrahydropyranyl and tetrahydrothiopyranyl group" such as tetrahydropyran-2-yl, 3- Sbromotetrahydropyran-2-yl, 4 -methoxytetrahydropyran-4-yl, tetrahydrothiopyran-2-yl S. and 4-methoxytetrahydrothiopyran-4-yl; "tetrahydrofuranyl and tetrahydrothiofuranyl group" such as tetrahydrofuran-2-yl and tetrahydrothiofuran-2-yl; "tri(lower alkyl)silyl group (hereinafter a lower alkyl moiety represents a group *selected from the group consisting of C C 6 alkyl group such as the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl and hexyl group) such as the trimethylsilyl, triethylsilyl, isopropyldimethylsilyl, tert-butyldimethylsilyl, S: diisopropylmethylsilyl, di(tert-butyl)methylsilyl and triisopropylsilyl group; "silyl group substituted with one or two aryl groups and two or one lower alkyl groups" such as diphenylmethylsilyl, diphenylbutylsilyl, diphenylisopropylsilyl, and diisopropylphenylsilyl; Doc: FP9907al.doc P814 8 5/FP-9907(PCT/tsa/gadjshicorrected pages of spedc/01/01 "lower alkoxymethyl group" (hereinaher an alkoxy moiety represents a group selected from the group consisting of C, C 6 alkoxy group such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy and hexyloxy), such as methoxymethyl, 1, 1 -dimethyl- 1 -methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, butoxymethyl and tert-butoxymethyl; "lower alkoxy-lower alkoxylmethyl group" such as the 2 -methoxyethoxymethyl group; "halogeno-lower-alkoxymethyl group" such as the 2,2,2-tr-ichloroethoxymethylI and bis(2-chloroethoxy)methyl group; "substituted ethyl group", for example an ethyl group substituted with a lower alkoxy group such as the I -ethoxyethyl or I -(isopropoxy)ethyl group, and for example a halogenoethyl group such as the 2,2,2-trichioroethyl group; "aralkyl group" (aryl moiety is selected from the group consisting Of C 6 C1 4 aryl group such as phenyl, naphthyl, biphenyl, anthryl and phenanthryl group), for example a lower alkyl group substituted with I to 3 aryl groups such as benzyl, ot-naphthyl, naphthyl, diphenylmethyl, triphenylmethyl, a-naphthyldiphenylmethyl and 9anthrylmethyl, and for exa mple a lower alkyl group substituted with I to 3 aryl groups, which are substituted with lower alkyl, lower alkoxy, nitro, halogen or cyano group, such as the 4-methylbenzyl, 2,4,6-trimethylbenzyl, 3,4,5-trimethylbenzy), 4-methoxybenzy], 4methoxyphenyldiphenylmethyl, 2-nitrobenzyl, 4-nitrobenzy], 4-chlorobenzyl, 4- ~:bromobenzyl and 4-cyanobenzyl group; alkoxycarbonyl group", for example lower alkoxycarbonyl group such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl and isobutoxycarbonyl, and for 0 0 0example lower alkoxycarbonyl group substituted with halogen or tri(lower alkyl)silyl group such as 2,2,2-tnichloroethoxycarbonyl and 2 -trimethylsilylethoxvcarbonyl, "alkenyloxycarbonyl group" (said alkenyl moiety is a C 2
C
6 alkenyl group) such as the vinyloxycarbonyl and allyloxycarbonyl group; and ~"aralkyloxycarbonyl group in which the aryl ring'is optionally substituted with one or Vooootwo lower alkoxy or nitro groups" such as the benzyloxycarbonyl, 4- 000000methoxybenzyloxycarbonyl, 3 4 -dimethoxybenizyloxycarbonyl, 2sooo nitrobenzyloxycarbonyl and 4 -nitrobenzyloxycarbonyl group.
:o 0. 1.: Doc: FP9907al-doc Doc: P99O~~doc P1485/FP-9907(PCT)Itsafgad/shlcorected pages of specIO3IOIIOI A preferable "general protecting group of hydroxyv group" is the tetrahvdropvranyl.
tetrahydrothiopyranyl. silvl. aralkvl,, or aralkvloxvcarbonyl group.
A more preferable "general protecting group of hydroxy group" is the tetrahvdropyran-2-vl. 4-methoxvtetrahv drop~rran-4-vlI. tetrahydrothiopyran-2-I.
diphenvlmethylsi lyi. benzyl. diphenylmethyl. triphenylmethyl. 4-methylbenzyL. 4methoxNvbenzvl. 2-nitrobenzN-l. 4-nitrobenzvl. 4-chlorobenzNvl. benzvloxv carbon vl. 4methoxybenzvl oxvc arbonvi. 2-nitrobenzvloxycarbonyl or 4 -nitrobenzyloxycarbon\
I
group.
A most preferable "general protecting group of hydroxy group' is the trimethylsilyl. tert-butyldimethylsilyl. tri phenylmethyl. benzvl or 4-methoxybenzyl group.
hvdroxv prtciggroup which can be removed by biological method such as hydrolysis in vivo is selected from the group consisting of "I -aliphatic acvloxy low.er alkyl group" (hereinafter. acyl moiety is selected from the group consisting of C, C 1 0 straight or branched chain alkanoyi group) such as form vlox ymethyl. acetoxymethyl. dimethylaminoacetoxymethyl.
propionvloxymethyl. butryloxymethyl. pivalo\-loxNvmethvl. valeryloxymethyl, isovalervioxvmethvl. hexanovioxvimrethvl. 1 -for-mx'oxvethvl. I -acetoxvethvl. I propionyloxyethyl. I -butyrylox-vethyl. I -pivalo\vloxvethvl. 1 -v.alervl:oxveth\:l. 1isovalervloxyethyl. I -hexanovloxyethyl. I -formrvlo xypropyl. l-acetoxypropyl. Ipropionyioxypropyl, I -butyryl oxypropvl. I -pivalovloxypropyl. I -valerv lox ypropyl. Iisovalervyloxypropyl, I -hexanoyloxypropyl. I -acetoxvbutyl. I -propionyloxybutyl. I butx'ryloxybutyl. I -pivaloyloxybutyl. 1-acetoxy'penty'l, I -propionyloxypentyl. I butyryloxypentyl. 1 -pivaloy lox ypenty I and I -pivaloyloxyhexyl: "1 -(aliphatic-acylthio)-(lower alkyl)group" such as form vlthi ometh yl.
acetylthiomethyl. dimethylami noacetvlthiomethvl. propionvithiomethyl.
butvrlthiomethyl, pivaloylthiomethyl. valervlthio methyl. isovalervlthiomethyl.
hexanoylthiomethyl, 1-formylthioethyl. 1-acetylthioethyl, l-propionylthioethyl.
I-
butyrvlthioethyl, I -pivaloylthioethvl, 1 -valervithioethyl, 1 -isovalerylthioethyl, I hexanoylthioethyl, 1 -formyithiopropyl, I -acetyithiopropyl, I -propionylthiopropyl,
I-
butvrylthiopropyl, 1 -pivaloylthiopropyl, 1 -valerylthiopropyl, I -isovalerylthiopropyl, Doc: FP9907s i.doc P8! 4851FP-9907(PCTI/tsa-2ad-sh/Engjjsh tra.nslaaion of spec (pages 1 -32)/08. 12.00 1 -hexanoylIthiopropyl. 1-acetylthiobutyI. 1-propionyithioiburvi. I-butyrylthiobutvl. 1 pivalayithiobutyl. I -acetylthiopentyl. I -propionylthiopentyl, 1 -buqTyylthiopentyl. 1 pivalovlthiopentyl and I -pivalovlthiohexyl: "I -(cvcloalkvlcarbonyloxv)-(low,,er alkyl.) group" such as the cyc lopentylcarbonyloxymethyl. cvc lohexvlcarbonyloxv~meth\vl. I cyc lopentylcarbonvloxyethyl. I clohexN lcarbonvlIoxveth\vL I cvclopentvlcarbonvloxvpropvl1. I -cvc lohex\vlcarbonv-loxv-propvL1 1cyc Iopentylcarbonvloxybutyl and I -cN-clohexvlcarbon\vloxvbutvl group: -aromatic acx'loxv)-(loNer alkyl) group (the aromatic acyl moiety is selected from the group consisting Of C 6 C 10 arv-icarbonyl groups)" such as the benzollox-vmethvl group.
"I -(lower alkox-vcarbonyl-o xv low.er alkyl) group" such as methoxvcarbonvloxvmethvl. ethoxycarbonyloxym ethyl. propoxycarbonvloxvm-ethyl.
isopropoxvcarbon\vloxy-methylI. butoxyc arbonyloxyml-ethyl.
isobutox-vcarbonvlIoxvmethvl,. pent\vlox\vcarbon\vloxv-meth\vl.
hex N- ox vc arbonv'loxymethyl. I -(methoxy-carbony loxy-)ethyl., I- (ethoxycarbonNyloxv)ethvl. I propox,.carbon\yIox\ )ethylI. I (isopropoxvcarbonyloxy)ethyl. I -butox vcarbonvlIox \)ethvl1. 1 (isobutox\vcarbonvloxv,)ethyl,. I -(tert-butox\ycarbon\vloxy-)ethvl. I- (pentvloxvcarbonvloxv)ethvl. hexvlIoxvcarbon\vlox%-)ethvl. I (methoxvcarbonyloxy)propyl. I -(ethoxycarbonylIo xv)propyl1. I- (propoxvcarbonvloxv)propyl. 1 isopropoxv-carbonvlIoxv-)propvl1. I (butoxycarbonyloxy)propyl. I (sobutox ycarbony I oxyvpropy 1. 1- (pentvloxvcarbonyloxx,)propyl1. I lox ycarbony Ioxy)propyl. I (methoxvcarbonvloxy)butyl. I -(ethoxvcarbonvloxv)butvl. I- (propoxvcarbonyloxy)butyl. I isopropoxvcarbonxlIox\y)butylI. I- (butoxvcarbonvioxy)butyl. I iosobutox vcarbon\ loxv-)butvll. I- (methoxvcarbonyloxv)pentNl. ethoxvcarbonyilo xy)pentvNl, 1I- (methoxvcarbonvloxv)hexvl and I -(ethoxvlcarbony loxv,)hexvl:.
"I1-(cvc loalkyloxycarbony lox ower alkyl) group" such as cvclopentyloxycarbonvloxymethvl. cyc lohexvloxvcarbonyloxvmethvI. 1I- (cyclopentyloxvcarbonyloxy)ethyl. I -(cyclohexvloxvcarbonyloxy)ethyl. I- (cyclopentyloxvcarbonyloxv)propyl. I -(cyclohexyloxycarbonyloxy)propyl. I (cvclopentvloxycarbonyloxv.)butyl. I -(cyclohexvloxycarbonyloxy)butyl.I- Doc: FP9907sldoc P8l48S/FP-99O7(PCT /tsa-2ad-shEnelish translation of spec (pages 1-32)/08.12.00 (cvclopentvloxvcarbonvloxv,)pentvl. 1 -(cyclohexyloxvcarbonyloxv)pentvlI, 1I- (cyclopentvloxycarbonyloxv)hexyl and I -(cyclohexyloxycarbonyloxy)hexvl; "phthalidvl group" such as the phthaldyl. dimethylphthal idyl and dimethoxyphthal idyl group: "oxodioxolenvlmethvl group" such as (5-phenN-l-2-oxo- 1 .3 -dioxolen-4-vl )methvl.
[5-(4-methvlIphenvl)-2-oxo- I.3)-dioxolen-4-vl,]methvlI. [5-(4-methoxvphenvl )-2-oxo- 1 .3-dioxolen-4-vI~methvl. [5-(4-fluorophenvlI)-2-oxo-lI.3-dioxolen-4--I ]methvl1. chlorophenvl)-2-oxo-l1.3)-dioxolen-4-vlIjmethvlI. (2-oxo-l1.3-dioxolen-4-vl )methvl1. methvl1-2-oxo-lI.3'-dioxolen-4-vlI)methvl. (5-ethvl1-2-oxo-lI.3)-dioxolen-4-vl1)methvl1. propvl-2-oxo- 1.3 -dioxolen-4-vlI)methvl-. (5-isopropv l-2--oxo-1 .'-dioxolen-4-vlI)methvlI and (5-butvl1-2-oxo-l1.3-dioxolen-4-vl )methvl:, "carbamnoyi group": "carbamnoyl group substituted with one or two lower alkyl groups": 'lower alkvl-dithi oethvl- group" such as methvldithioethvl. eth-vldithioethvlI.
propyldithioethyl. butyldithioethy1. penty ldithioethy I and hexvldithioet h\l group: and "I -(c-o,,akNIx~abn- group" such as the pivalox'loxvmethv'loxy-carbonvl group.
A preferable "hvdroxy protecting group which can be remo-ved by bioloaical method such as hydrolysis in vivo' is selected from the group consisting of a I1- (aliphatic acvloxN)-(lower alkyl) group. a Il-(cvcloalkvlIcarbonvloxv-)-(lowAer alky I) group. a I -(lower alkox\vcarbonvlox%-)-( lower alkyl) group. a I- (cvcloalk\vloxvlcarbonv-loxv-( lOower alky l) group. a phthalidvl and an oxodioxolenvlmethyl group.
A more preferable "hvdrox\ protecting group which can be removed by biological method such as hydrolysis in vivo" is selected from the group consisting of acetoxymethyl, propionyloxymethyl. butyryloxym ethyl. pivaloyloxymethyl.
valeryloxymethyl. 1 -acetoxvethyl. but-vrvloxvethvl. I -pivalovlox\vethvlI.
cyclopentvlcarbonyloxvmeth\vl. cy-clohexNylcarbonvloxymethNy!. I cyclopentylcarbonyloxyethyl. 1 -cyclohexvlcarbonvlIoxvethyl.
methoxycarbonyloxyrmethyl. ethoxvcarbonvloxv.,methyl, propoxycarbonyloxymethyl, isopropoxycarbonyloxymethyl, butoxycarbonyloxymethyl, isobutoxycarbonyloxymnethyl, 1 -(methoxycarbonyloxy)ethyl,
I-
(ethoxycarbonyloxy)ethyl, I -(isopropoxvcarbonyloxy)ethyl, Doc: FP9907s]I doc PS 1485/FP-9907 PCT1/tsa-g2ad-shlEngljish translation of spec (pages 1-32)/08.12.00 cv ,clopent- l.oxvcarbonvlIoxv-methyL1 cy-clohexvloxycarbonvloxvmethvl. 1- (cvclopentvloxvcarbonyloxv)ethvl. 1 -(cyclohexyloxvlcarbonyloxv)ethyl, phthalidyl, (5-phenvl-2-oxo- 1.3 -dioxolen-4-vl )methfl. [5-(4-methylphenyl )-2-oxo-l1.3)-dioxolen- 4-vljmethvl. (5-methvl-2-oxo-lI.3)-dioxolen-4-vl,)m ethyl and (5-ethvl1-2-oxo-l.3dioxolen-4-vI)methvl group.
A most preferable "hydroxy protecting group which can be removed by biological method such as hydroly~sis in vivo" is selected from the group consisting of acetoxvmethvl. propionyloxvmethvl. butyryloxvmethvl. pivalovloxym ethyl.
valeryloxymethyl. c vclopentvlIcarbonvlIoxv-methvlI. cv7clohexylIcarbonvlIoxvmnethv.
methoxvcarbonvloxvm-ethvl. ethoxycarbonyloxvm ethyl. propoxycarbonv loxvymethyl.
isopropoxvcarbonv-loxvmethvlI. butoxycarbonvloxvmethvl.
isobutoxycarbonyloxvmethyl. cv-clopent\vloxv'carbonvlIoxv~methylI.
c-c lohex\yloxv~carbonylIoxv-met hyl. (5 -phenvl1-2-oxo-lI.3'-dioxolen-4-\l )methylI. methv lphenvl)-2--oxo- 1.3-dioxolen-4-yl I]methvlI. (5i-methvl-2-oxo-1I.3-dioxolen-4yl )methyll and (5-ethvl-2-oxo- I.'3-dioxolen-4-vl )methvlI group.
The term 'pharmaceuticallIy acceptable ester. ether and N-alkylcarbamovl deri-vatives" refers to a derivati-ve that is a useful medicament without significant toxicity.
The ester residue of ester derivatives is selected from the group consisting of "carbonvl and oxycarbonyl group to which a straight or branched chain C 1 alkyl group is attached". in which said alkyl group is selected from the group consisting of methyl. ethyl. propyl. isopropyl. butyl. isobutvl. sec-butyl. tert-butvl.
pentyl. isopentyl. 2-methylbutvl. neopentyl. Il-ethylpropyI. hex-vl. isohexvl1. 4methylpentyl. 3-methylpentyl. 2-methylpentvl. I -methylpentvl. 3.3-dimethylbutyl.
2.2-dimethvlbutyl. 1.1 -dimethylbutyl. 1 .2-dimrethy Ibutyl. I .'-dimethvlbutvl. 2.3dimethylbutvl. 2-ethvlbutNl. heptyl1. 1 -methylhexyl. 2-methylhexyl. 3- methvlh-exyl. 4methylhexvl. 5-methylhexyl. I -propylbutyl. 4.4-di methyl pentyv1. octyl. I methylheptyl. 2-methylheptylI. 3 -methyl heptyl, 4-methNvlheptyl. 5-methvlheptvl. 6methylheptyI. I -propyl pentyl. 2-ethyihexyl. 5.5-di methylhexyl. nonyl. 3)-methyloctyl, 4-methyloctyl, 5-methyloctyl. 6-methyloctvl, I -propyihexyl. 2-ethylh-eptyl. 6.6dimethyiheptyl, decyl, 1 -meth-vinonil, 3 -methylnonyl, 8-methylnonvl. 3-ethvloctvl.
3 .7-dimethyloctyl, 7,7-dimethilocti., undecyl, 4,8-dimethyinonyl, dodecyl. tride cyl, tetradecyl, pentadecyl, 3,7,11 -trimethyldodecyl, hexadecyl, 4,8,1 2-trimethvltridecyl, Doc: FP99O7sldoc P814851FP-9907(PCTI/tsa-gad-shlEnglish translation of spec (pages 1-32)0812.00 1 -methylpentadecyl, I 4-methyipentadecyl, 13,1 3-dimethyltetradecyl, heptadecyl, methyihexadecyl, octadecyl, I -methylheptadecyl, nonadecyl, icosyl, 3,7,1 1,15tetramethyihexadecyl and henicosyl groups; "carbonyl and oxycarbonyl group to which a straight or branched chain C 2
C
21 alkenyl or alkynyl group is attached", in which said alkenyl or alkynyl group is selected from the group consisting of ethenyl, I -propenyl, 2-propenyl, I -methyl-2-propenyl, I methyl- I -propenyl, 2-methyl- I -propenyl, 2-methyl-2-propcnyl, 2 -ethyl-2-propenyj, I1butenyl, 2-butenyl, I -methyl-2-butenyl, I -methyl- I -butenyl, 3-methyl-2-butenyl, I1-ethyl 2-butenyl, 3-butenyl, I -methyl-3-butenyl, 2-methyl-3-butenyl, I -ethyl-3-butenyl, I pentenyl, 2-pentenyl, I -methyl-2-pentenyl, 2-methyl-2-pentenyl, 3-pentenyl, I -methyl-3pentenyl, 2-methyl-3-pentenyl, 4-pentenyl, I -methyi-4-pentenyl, 2 -methyl-4-pentenyl, Ihexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, cis-8-heptadecenyl, cis, cis-8,1 I1heptadecadienyl, cis, cis, cis- 8, 11,1 4-heptadecatrienyl, cis- 1 0-nonadecenyl, and cis- 12icosenyl; "carbonyl and oxycarbonyl group to which a straight or branched chain C 2
C
2 1 alkenyl or alkynyl group is attached", in which said alkenyl or alkynyl group is selected from the group consisting of ethynyl, 2-propynyl, I-methyl-2-propynyl, 2-methyl-2propynyl, 2-ethyl-2-propynyl, 2-butynyl, I -methyl-2-butynyl, 2-methyl-2-butynyl, I ethyl-2-butynyl, 3 -butynyl, I -m ethyl -3 -butynyl, 2-methyl-3-butynyl, I -ethyl -3 -butynyl, 2-pentynyl, I -methyl-2-pentynyl, 2-methyl-2-pentynyl, 3-pentynyl, I -methyl-3-pentynyl, 2-methyl-3-pentynyl, 4-pentynyl, I -methyl-4-pentynyl, 2 -methyl-4-pentynyl, 2-hexynyl,.
3-hexynyl, 4-hexynyl and "carbonyl and oxycarbonyl group to which straight or branched chain C, C 2 1 alkyl :group which has one or more substituents selected from the group consisting of lower alkoxy, halogen (hereinafter for example fluorine, chlorine, bromine and iodine, preferably fluorine and chlorine) and nitro groups is attached", in which said substituted alkyl group is selected from the group consisting of methoxymethyl, ethoxymethyl, methoxyethyl, ethoxyethyl, trifluoromethyl, trichloromethyl, difluoromethyl, dichloromethyl, dibromomethyl, fluoromethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, 2-bromoethyl, 2-chloroethyl, 2-fluoroethyl, 2-iodoethyl, 3-chloropropyl, 4-fluorobutyl, 6iodohexyl, 2,2-dibromoethyl, nitromethyl, dinitromethyl, I -nitroethyl, 2-nitroethyl and I ,2-dinitroethyl; "carbonyl and oxycarbonyl group to which a (C 6 CIO aryl)-(Ci C 2 1) alkyl group wherein said aryl moiety optionally has one or more substituents selected from the Doc: FP9907al-doc Doc:FP99~a Idoc P 14 85IFP-9907(PC Ta/Vgad/sh/coffected pages of spec/03/0 1/01 group consisting of lower alkyl. lower alkoxy. halo and nitro groups is attached". in which said arvialkyl group is selected from the group consisting of benzyl. anaphthxvlmethvl. 1-naphthvlmeth%?l. inden--Imethvl. pheniarnhrenylmethyl.
anthracenylm ethyl, diphenvlmethyl. tripheny'lmethyl. I -phenethyl. 2-phenethyl. I naphthylethx'l. 2-naphthylethyl. I -phenNlpropyl. 2-phenvlpropyl. 3 -phenylpropyl. I naphthylpropyl. 2-naphthylpropx'l. 3 -naphthylpropyl. I -phenyl butyl. 2-phenylIbutyl. 3phenylbut-yl, 4-phenylbutyl. I -naphthylbutyl. 2-naphthx'lbutyl. 3-naphthylbutyl. 4naphthylbutyl. I -phenylpentyl. 2-phenyl pentyl. 3-phenylpentx'l. 4-phenyl penty 1. phenylpent-vl. 1 -naphthylpentyl. 2 -naphthylpentyl. 3 -naphthylpentyl. 4naphthylpentyl. 5-naphthylpentyl. 1 -phenylhexyl. 2-pheny'lhexyl. 3-phenylhexyl. 4phenvlhexyl. 5-phenylhexyl. 6-phenylhexyl. I -naphthylhexyl. 2-naphthylhexyl. 3naphthylhexyl. 4-naphthylhexyl. 5-naphthylhexyl and 6-naphthylhexyl: 'carbonvl and oxycarbonyl group to wvhich a C 6 CIO aryl group which optionally has one or more substituents selected from the group consisting of lower alkyl. lower alkox-v. halo and nitro groups is attached", in which said aryl group is selected from the group consisting of phenyl. naphthvl. 2-fluorophenyl. 3-fluorophenyl. 4fluorophenyl. 2-chlorophenyl. 3-chlorophenyl, 4-chlorophenyl. 2-bromophenyl. 3bromophenyl, 4-bromophenyl. 3.5-difluorophenyl. 2.5-difluorophenyl. 2.6difluorophenyl. 2.4-d ifluorophenl. 3.5-dibromophenyl. 2.5-dibromophenyl. 2.6dichlorophenyi. 2.4-dichlorophenyl. 2.3 .6-trifluorophenyl. 2.3.4-trifluorophenvl.
3.4.5-trifluorophenyl. 2.5.6-trifluorophenyl. 2.4.6-trifluorophenyl. 2.3.6tribromophenyl. 2.3.4-tribromophenyl. 3 .4.5-tribromophenyl. 2.5.6-trichlorophenyl.
2.4.6-trichlorophen%.l. 1 -fluoro-2-naphthyl. 2-fluoro- I-naphthvl. 3-fluoro- i-naphthyl.
I -chiloro-2 -naphthyl. 2-chloro- I -naphthyl. 3-brorno- I -naphthyl. 3 .8-difluoro- I1naphthvl. 2.3-difluoro-l1-naphthyl. 4.8-difluoro-lI-naphthyl. 5.6-difluoro-lI-naphthyl.
3.8-dichloro- I-naphthyl. 2.3-dichloro- I -naphthyl. 4.8-dibromo- I -naphthyl. 5.6dibromo- 1 -naphthvl. 2.3 .6-trifluoro- I -naphthNvl. 2.3.4-trifluoro- I -naphthyl. 3.4.5trifluoro- I -naphthyl, 4.5.6-trifluoro- I -naphthyl. 2.4.8-trifluoro- I -naphthyl. 2methylphenyl, 3-methylphenyl. 4-methylphenyl. 2-ethylphenvyl. 3-propylphenyl. 4ethvlphenvl. 2-butylphenyl. 3-pentx'lphenyl. 4-pentylphenyl. 3.5-dimethylphenx'l. dimethylphenyl, 2,6-dimethylphenvl, 2.4-dimethvl phenyl. 3),5-dibutylphenyl, dipentylphenyl, 2,6-dipropylmethylphenyl, 2,4-dipropylphenyl, 2,3,6trimethylphenyl, 2.3 .4-trimethylphenvyl, 3 .4.5-tr-imethyiphenyl, 2,5 ,6-trimethylphenyl, Doc: FP9907s I.doc P81 485/FP-9907( PCT)/tsa-gad-shlEngplish translation of spec (pages 1-32)/08 12 00 12 2.4.6-trimethylphenyl. 2.3 .6-tr-ibutylphenyl. 2.3.4-tripentylphenyl. 3.4.5 tributyiphenyl, 2.5,6-tripropylmethyilphenyl. 2.4.6-tripropylphenyl, 1 -methvl-2naphthyl. 2-methyl-I -naphthyl. 3-methyl-i -naphthyl. I -ethyl -2 -naphthyl. 2-propyl- 1naphthyl, 3-butyl-l1-naphthyl. 3 .8-dimethx'l- I-naphthyl. 2.3-dimethyl- I-naphthyl. 4.8dimethxl- 1-naphthy'l. 5.6-dimethyl-l-naphthyl. 3 .8-diethvl-l1-naphthyl. 2.3 -dipropyl 1 -naphthyl. 4.8-dipentvl- 1 -naphthyl. 5.6-dibutx'l- 1 -naphthyl. 2.3 .6-trimethyl- I1naphthyl. 2.3,4-trimethyl-1 -naphthyl. 3.4.5-trimethyl-lI-naphthyl. 4.5.6-trimethv- Inaphthyl. 2.4.8-trimethyl-lI-naphthyl. 2- methoxyphenyl. 3- methoxyphenyl. 4methoxyphenyl. 2-ethoxyphenyl. 3-propoxyphenvl. 4-ethoxNyphen\vl. 2--butoxyphenyl.
-3-pentyloxyphenyl. 4-pentx'loxyphenyl. 3.5-dimethoxyphenyl. 2.6-dimethoxvphenvl. 2.4-dimethoxyphenyl. 3.5-dibutoxyphenyl. dipentvloxyphenyl. 2.6-dipropoxymethoxyphenyl. 2.4-dipropoxyphenyl. 2.3.6trimethoxyphenvyl. 2.3.4-trimethoxyphenyl. 3.4.5-trimethoxyphenyl. 2.5.6trimethoxyphenyl. 2.4.6-trimethox\vphenyl. 2.3 .6-tributoxyphenyl. 2.3.4tripentyloxyphenyl. 3.4.5-tributoxyphenyl. 2.5.6-tripropoxyphenyl. 2.4.6tripropoxyphenyl, I -methoxN-2--naphth\-l. 2-methoxy- I-naphthyl. 3-methoxy-lInaphthyl. 1-ethoxv-2-naphthyl. 2-propoxy-l-naphthyl. 3-buboxv-l-naphthyl. 3.8dimethoxy- I-naphthyl. 2.3-dimethoxy-l1-naphthyl. 4.8-dimethoxy-lI-naphthyl. 5.6dimethoxy-l1-naphthyl, 3.8-diethoxy-lI-naphthyl. 2.3-dipropoxy-l1-naphthyl. 4.8dipentx'loxy-1-naphthyl. 5.6-dibutox I-naphthyl. 2.3).6-trimethox\---naphthv.l. 2.3,4trimethoxv-lI-naphth,,l. 3.4.5-trimethoxv-1I-naphthyl. 4.5.6-trimethoxv -1-naphthNvl.
2.4.8-trimethoxy-lI- naphthyl. 2-nitrophenyl. 3-nitrophenyi. 4-nitrophenyl. dinitrophenyl. 2.5-dinitrophenyl. 2.6-dinitrophenyi. 2.4-dinitrophenvl. 2.3.6trinitrophenyi, 2.3.4-trinitrophenv-I. 3.4.5-trinitrophenyi. 2.5.6-trinitrophen-vl. 2.4.6trinitrophenyl. I -nitro-2-naphthyl. 2-nitro- I-naphthyl. 3-nitro- I -naphthyl. 3).8-dinitro- I -naphthyl. 2.3-dinitro- I -naphthyl. 4.8-dinitro- I -naphthyl. 5.6-dinitro- I -naphthv.
2.3 .6-trinitro- 1 -naphthyl, 2.3 .4-tri nitro- I -naphthyl. 3).4.5-trinitro- I -naphthyl. 4.5.6trinitro- I-naphthyl and 2.4.8-trinitro-1I-naphthyl: "carboxy (CI C 10 )alkvlcarbonyl group" such as succinoyl.
glutaroyl, and adipoyl; "residue of salt of a phosphate diester which independently has two lower alkyl groups', and Doc: FP9907slI.doc P81 485/FP-9907(PCT)/tsa-ead-shi/English translation of spec (pages 1 -32)/08. 12.00 13 "residue forming ester of amino acid which is optionally protected with a tertbutyloxycarbonyl, benzyloxycarbonyl or trityl group" such as glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, glutamine and glutamic acid.
A preferable ester residue of ester derivatives is R 6 CO- or R 6 OCO- group wherein R 6 is selected from the group consisting of hydrogen; a Ci C 2 1 alkyl group; a C 2
C
21 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a C 1 C21 alkyl group substituted with 1 to 4 substituents selected from the group consisting of lower alkoxy, halo and nitro groups; a C, C21 alkyl group substituted with I to 3 C 6 CIO aryl groups which are optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups; and a C 6 CIO aryl group which is optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo, and nitro groups.
A more preferable ester residue of ester derivatives is R 6 CO- or R 6 OCO- group wherein R 6 is selected from the group consisting of hydrogen; a C, Cz2 alkyl group; a C 2
C
2 1 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a Ci C 6 alkyl group substituted with 1 to 4 substituents selected from the group consisting of C, C 4 alkoxy, halo and nitro groups; a C 1
C
6 alkyl group substituted with 1 to 3 C 6 Cio aryl groups which are optionally substituted with I to 3 substituents selected from the group consisting of C, C 4 alkyl, CI C4 alkoxy, halo and nitro groups; and a C 6 CIO aryl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of Cl C4 alkyl, CI C4 alkoxy, halo and nitro groups.
A more preferable ester residue of ester derivatives is R6CO- or R 6 OCO- group *wherein
R
6 is selected from the group consisting of a Ci C 2 1 alkyl group; a C6 C 20 S: alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group substituted with one substituent selected from the group consisting of C C 4 alkoxy and nitro groups; a Ci C 6 alkyl group substituted with 1 to 3 substituents selected from the group consisting of halogen; a C, C 4 alkyl group substituted with 1 to 3 phenyl or naphthyl groups which are optionally substituted with I to 3 substituents selected from the group consisting ofC, C 4 alkyl, C, C 4 alkoxy, halo S. and nitro groups; and a phenyl or naphthyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of C C 4 alkyl, C, C 4 alkoxy, halo and nitro groups.
Doc: FP9907al.doc P81485/FP-9907(PCT)/tsagad/sh/corrected pages of spcc/03/01/01 A more preferable ester residue of ester derivatives is R 6 CO- or R60CO- group wherein R 6 is selected from the group consisting of a C 6
C
20 alkyl group; a Clo C 20 alkenyl group having 1 to 3 double bonds: a C 3 Cs alkvnyl group having one triple bond; a Ci C 4 alkyl group substituted with one substituent selected from the group consisting ofCi C 4 alkoxy and nitro groups: a Ci C 4 alkyl group substituted with I to 3 substituents selected from the group consisting of fluoro and chloro groups: a Ci
C
4 alkyl group substituted with 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of C C, alkvl. CI C 4 alkoxy. fluoro and chloro groups: and a phenyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting ofCi C: alkvl. Ci C 4 alkoxy. fluoro and chloro groups.
A more preferable ester residue of ester derivatives is R 6 CO- or R"OCO- group wherein R 6 is selected from the group consisting of a C 6
C
20 alkyl group: a CIW C 2 0 alkenyl group having 1 to 3 double bonds: a C3 C; alkynyl group having one triple bond: a Ci C 4 alkyl group substituted with one substituent selected from the group consisting ofCI C 4 alkoxy. fluoro. chloro and nitro groups: a Ci C 4 alkvl group substituted with 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting ofC, C 2 alkvl. Ci C 4 alkoxv. fluoro and chloro groups; and a phenyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of Ci C alkvl. C C 4 alkoxv. fluoro and chloro groups.
A still more preferable ester residue of ester derivatives is R 6 CO- or R 6
OCO-
group wherein R 6 is selected from the group consisting of a C 6
C
2 alkyl group: a Cio
C
2 0 alkenyl group having 1 to 3 double bonds: a C 3 C; alkvnyl group having one triple bond; a C, C 4 alkyl group substituted with one substituent selected from the group consisting of Ci C 4 alkoxy groups: and a C C 4 alkvl group substituted with 1 or 2 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting ofCi C, alkvl. Ci C 4 alkoxy. fluoro and chloro groups.
A most preferable ester residue of ester derivatives is R 6 CO- or R 6 OCO- group wherein R 6 is selected from the group consisting of a C 6
C
20 alkyl group and a Clo
C
20 alkenyl group having 1 to 3 double bonds.
An ether residue of ether derivatives is selected from the group consisting of Doc: FP9907sl.doc P81485/FP-9907(PCT /tsa-gad-sh/English translation of spec (pages 1-32)/08.12 00 "straight or branched chain C, C 21 alkyl group" such as the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, neopentyl, I -ethyipropyl, hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2methylpentyl, I -methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1, 1 -dimethylbutyl, 1 ,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, heptyl, I1methyihexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methyihexyl, I propylbutyl, 4,4-dimetbylpentyl, octyl, I -methyiheptyl, 2-methyiheptyl, 3-methyiheptyl, 4-methyiheptyl, 5-methyiheptyl, 6-methyiheptyl, I -propylpentyl, 2-ethylhexyl, dimethyihexyl, nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6-methyloctyl, I1propylbexyl, 2-ethyiheptyl, 6,6-dimethylheptyl, decyl, I -methylnonyl, 3-methylnonyl, 8methylnonyl, 3-ethyloctyl, 3,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8dimethylnonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,1 1-trimethyidodecyl, hexadecyl, 4,8,1 2-trirnethyltridecyl, I -methylpentadecyl, I 4-methylpentadecyl, 13,13dimethyltetradecyl, heptadecyl, 1 5-rnethylhexadecyl, octadecyl, I -methyiheptadecyl, nonadecyl, icosyl, 3,7,11,1 5-tetramethylhexadecyl and henicosyl groups; "straight or branched chain C 2
C
2 alkenyl or alkynyl group" such as ethenyl, I1propenyl, 2-propenyl, 1-methyl-2-propenyl, I -methyl-1-propenyl, 2-methyl-I -propenyl, 2-methyl-2-propenyl, 2-ethyl-2-propenyl, I -butenyl, 2-butenyl, 1 -methyl-2-butenyl, I1methyl-I -butenyl, 3-methyl-2-butenyl, I -ethyl -2-butenyl, 3-butenyl, I -methyl-3-butenyl, **:2-methyl-3-butenyl, I -ethyl-3-butenyl, I -pentenyl, 2-pentenyl, I -methyl-2-pentenyl, 2- :0:.:.methyl-2-pentenyl, 3-pentenyl, I -methyl-3-pentenyl, 2-methyl-3-pentenyl, 4-pentenyl, I1methyl-4-pentenyl, 2-methyl-4-pentenyl, 1 -hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, hexenyl, cis-8-heptadecenyl, cis, cis-8,1 1-heptadecadienyl, cis, cis, cis-8,I 1,14- 0heptadecatrienyl, cis-lIO-nonadecenyl, cis-1I2-icosenyl, ethynyl, 2-propynyl, Il-methyl-2propynyl, 2-metbyl-2-propynyl, 2-ethyl-2-propynyl, 2-butynyl, I -methyl-2-butynyl, 2methyl-2-butynyl, I -ethyl-2-butynyl, 3-butynyl, I -methyl-3-butynyl, 2-methyl-3-butynyl, *00* *I -ethyl-3-butynyl, 2-pentynyl, I -methyl-2-pentynyl, 2-methyl-2-pentynyl, 3-pentynyl, I1- *0000*methyl-3-pentynyl, 2-methyl-3-pentynyl, 4-pentynyl, I -methyl-4-pentynyl, 2-methyl-4- V0:: pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl and 000"straight or branched chai n C I C 2 alkyl group which has one or more substituents :0 selected from the group consisting of lower alkoxy, halogen (hereinafter for example fluorine, chlorine, bromine and iodine, preferably fluorine and chlorine) Doc: FP9907al-doc Doe: P99O~~doc PI 485/FP-99O7(PCT)/tsa/gadtsh/coriected pages of spec/03IOI/OI and nitro groups" such as methoxymethyl, ethoxymethyl. methoxvethyl. ethoxyethyl.
trifluoromethyl. trichioromethyl. difluoromethyl, dichioromethyl, dibromomethyl.
fluoromethvll. 2.2.2-trifluoroethvl. 2.2.2-trichloroethNl. 2-bromoethNvl. 2-chloroethvl.
2-fluoroethyl. 2-iodoethyl. 3 -chloropropyl. 4-fluorobutyL. 6-jodohexyl. 2.2di bromoethyl. nitromethyl. djnitromethyl. I -nirroethyl. 2-nitroethyl and 1.2dinitroethyl:
"(C
6
C
10 )arvl-(C 1
C
2 1 I)alkN-l group wherein said aryl moiety optionally has one or more substituents selected from the group consisting of lower alkyl. lower alkoxy.
halo and nitro group" such as benzvl. ct-naphthylmethyl. j3-naphthylmethyl.
inden Im ethyl. phenanthrenylmethyl. anthracenylmethvl. diphenyl methyl, triphenvlmethvl, 1 -phenethyl. 2-phenethyl. I -naphthylethyl. 2-naphthylethyl. 1phenvlpropyl. 2-phen\'lpropyl. 3-phenylpropyl. I -naphthx'lpropyl. 2-naphthylpropyl, 3naphthvlpropyl, 1 -phenylbutyl. 2-phenvlbutyl. 3)-phenylbutN'I. 4-phenylbutNl 1naphthylbutyl. 2-naphthylbutyl. 3-naphthylbutvyl, 4-naphth\'lbutvl. I -phenylpentyl. 2phenylpentyl. 3-phenylpentyl. 4-phenylpentyl. 5 -phenylpentyl. I -naphthx'l penty 1. 2naphthylpentyl. 3 -naphthylpentvl. 4-naphthylIpentvl. 5 -naphthyl penrvl1. 1 -phenx'lhexyl.
2-phenvlhexvl. 3-phenyihexyl. 4-phenylhexyl. 5-phenyihexyl. 6-pheny'lhexyl. Inaphthvlhexvl. 2-naphthylhexNvl. 3-naphthx'lhexyl. 4-naphthylhexyl. and 6-naphthvlhexyl, and
"C
6 CIO arvi group which optionally has one or more substituents selected from the group consisting of lower alkyl. lower alkoxv. halo and nitro groups" such as phen-vi. naphthyl. 2-fluorophenyl. 3-fluorophenyi. 4-fluorophenyl. 2-c hlorophenvi. 3chlorophenvl. 4-chlorophenyl. 2-bromophenyl. 3-bromophenyl. 4-bromophenvl. difluorophenyl. 2.5-difluorophenyi ilrpev. 2.4-difluorophenyl. dibromophenyi. 2,5-dibromophenyi. 2.6-dichlorophenyl. 2.4-dichlorophenyi. 2.3.6trit fluorophenvl. 2.3.4-trifluorophen .4.5-trifluorophenvl. 2.5.6-trifluorophenvl.
2.4.6-trifluorophenyl. 2.3,6-tribromophenyvi 2.3 4-tribromophenyl. 3.4.5tribromophenyl, 2,5,6-trichiorophenyl. 2.4.6-trichlorophenyl. I -fluoro-2-naphthyl. 2fluoro- 1-naphthyl, 3-fluoro- I-naphthN'l, I -chloro-2-naphthyl. 2-chloro-lI-naphthyl, 3bromo-1 -naphthyl, 3,8-difluoro- 1-naphthyl. 2.3-difluoro-lI-naphthvl. 4.8-difluoro- 1naphthyl, 5.6-difluoro-1 -naphthyl. 3.8-dichioro-lI-naphthyl, 2,3-dichioro-lI-naphthyl, 4.8-dibromo-lI-naphthyl, S ,6-dibromo- I-naphthyl, 2,3 ,6-trifluoro- 1-naphthyl, 2.3.4trifluoro-lI-naphthyl, 3 ,4.5-trifluoro-lI-naphthyl, 4,5,6-trifluoro-lI-naphthyl, 2,4.8- Doc: FP9907s].doc P81485/FP-9907(PCT)/tsa-g2ad-shlEngl]ish translation of spec (pages 1-32)/08.12.00 trifluoro- 1 -naphthyl, 2-methyiphenyl. 3-methylphenyl. 4-methylphenyl, 2ethylphenyl. 3-propyiphenyl, 4-ethyiphenyl, 2-butyiphenyl, 3-pentylphenyl. 4pentyiphenyl. 3 .5-dimethylphenyl. 2.5-dimethyiphenyl. 2.6-dimethylphenNiI. 2.4dimethylphenyl, 3 .5-dibutylpheny'l. 2.5 -dipentylphenx'l. 2.6-dipropylmethylphenv.
2.4-dipropylphenvl. 2,3 .6-trimethylphenyl. 2.3).4-trimethylphenyl. 3.4.5trimethylphenyl. 2.5,6-trimethyiphenyl. 21.4.6-trimethylphenNII. 2.3.6-tributylphenyl.
.4-tri pent\'lphenyl, 3.4.5-tributylphenyl. 2.5.6-tripropylmethN lphenyl. 2.4.6tripropyiphenyl. 1 -m ethyl -2 -naphthyl. 2-methyl- I -naphthyl. 3-methyl- I -naphthyl. I ethyv1-2-naphthyl. 2-propyl- 1 -naphthyl. 3-butvl- I -naphthyl. 3.8-dimethyl- I-naphthyl.
2.3-dimethyl- I -naphthyl. 4.8-dimethyl- I -naphthyl. 5.6-dimethyl- I -naphthyl. 3.8diethvl-1I- naphthyl. 2.3-dipropvl- 1 -naphthyl. 4.8-dipentyl- I -naphthyl. 5.6-dibutyl- I naphthyl. 2.3 .6-trimethyl- I -naphthyvl. 2.3.4-trimethyl- I -naphthyl. 3.4.5-trimethyl- I naphthyl1. 4.5.6-trimethyl- I -naphihyl. 2.4.8-trimethyl- I -naphthyi. 2-methoxyphenyl.
3-methoxyphenyl. 4-methoxyphenyi. 2-ethoxNvphenvl-. 3-propoxyphenyl. 4ethoxyphenyi. 2-butoxyphenyi. 3-pentoxyphenyl. 4-penryloxyphenyl. dimethoxyphenyi. 2.5-dimethoxyphenyl. 2.6-dimethoxyphenyl. 2.4-dimethoxyphenyl.
2.5-dipentyloxyphenyl. 2.6-dipropoxymethoxyphenyl. 2.4dipropoxyphenvi, 2.3.6-trimethoxyphenyl. 2 .3.4-trimethoxyphenyl. 3.4.5 trimethoxyphenyi. 2.5.6-trimethoxyphenyl. 2.4.6-trimethoxyphenyl. 2.3.6ibutoxvphenvl. 2.3,4-tripentyloxyphenyl. 3.4.S-tributoxyphenvl. 2.5.6tripropoxyphenvl. 2.4.6-tripropoxyphenyl. 1 -methoxv-2-naphthyl. 2-methoxv- I1naphthyl. 3-methoxv- I-naphthyl. I1-ethoxy-2-naphthyl. 12-propoxv- I-naphthyl. 3butoxy- I-naphthyl. 3.8-dimethoxy- I-naphthvl. 2.3-dimethoxv- I-naphthyi. 4.8dimethoxv-I -naphthyl. 5.6-dimethoxy- I-naphthyl. 3.8-diethoxy- I-naphthyl. 2.3dipropoxy- I -naphthyl. 4.8-dipentyloxy- I -naphthyl. 5.6-dibutoxy- I -na phthyl. 2.3.6trimethoxv- 1 -naphthvl. 2.3.4-trimethoxv- I -naphthyI. 3 .4.5-trimethox\ I -naphthyl.
4.5.6-trimethoxy- 1 -naphrhyl. 2.4.8-trimethoxy- I -n aphthyl. 2-nitrophenyl. 3nitrophenyl, 4-nitrophenyl. 3.5-dinitrophenyi. 2.5-dinitrophenyi. 2.6-dinitrophenyl 2.4-dinitrophenyt. 2.3.6-trinitrophenyl. 2.3 .4-trinitrophenyl. 3.4.5-trinitrophenyi.
2,5.6-trinitrophenyi, 2.4.6-trinirrophenyi. I -nitro- 2-naphthyl. 2-nitro- I-naphthyl. 3 nitro- 1 -naphthyl, 3 .8-dinitro- 1 -naphthyl. 2,3-dinitro- I -naphthyl, 4.8-dinitro- I naphthyl, 5,6-dinitro- 1 -naphthyl. 2.3 .6-trinitro- I -naphthyl, 2,3 ,4-trinitro- I -naphthyl, 3 A4,5-tr-initro- 1 -naphthyl, 4,5 .6-tri nitro- I -naphthyl and 2,4,8-tinitro- 1 -naphthyl.
Doc: FP9907s I.doc P8 1485/FP-9907(PCT)/rsa-gPad-shlEngplish translaion of spec (pages 1-32)/08.12.00 A preferable ether residue of ether derivatives is selected from the group consisting of a CI C 2 alkyl group; a C 2
C
2 1 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a C, C21 alkyl group which has 1 to 4 substituents selected from the group consisting of lower alkoxy, halo and nitro groups; a C, C 2 alkyl group which has 1 to 3
C
6 CIo aryl groups which are optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups; and a C 6 Clo aryl group which is optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups.
A more preferable ether residue of ether derivatives is selected from the group consisting of a C, C 2 1 alkyl group; a C 2
C
2 1 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group which has I to 4 substituents selected from the group consisting of CI C 4 alkoxy, halogen and nitro groups; a Ci C 6 alkyl group which has I to 3 C 6 Cno aryl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of Ci C 4 alkyl, C,
C
4 alkoxy, halo and nitro groups; and a C 6 CIO aryl group which is optionally substituted with I to 3 substituents selected from the group consisting of Ci C 4 alkyl, C,
C
4 alkoxy, halo and nitro groups.
A more preferable ether residue of ether derivatives is selected from the group consisting of a C, C 2 alkyl group; a C 6 C2o alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group which has one Ssubstituent selected from the group consisting of C C 4 alkoxy and nitro groups; a C,
C
6 alkyl group which has 1 to 3 substituents selected from the group consisting of halo groups; a C, C 4 alkyl group which has 1 to 3 phenyl or naphthyl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of C,
C
4 alkyl, Ci C 4 alkoxy, halo and nitro groups; and a phenyl or naphthyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of CI
C
4 alkyl, Ci C 4 alkoxy, halo and nitro groups.
A more preferable ether residue of ether derivatives is selected from the group consisting ofa C 6
C
20 alkyl group; a Clo C 20 alkenyl group having 1 to 3 double bonds; a C3 C 5 alkynyl group having one triple bond; a C, C 4 alkyl group which has one substituent selected from the group consisting of C C 4 alkoxy and nitro group; a Ci C 4 alkyl group which has 1 to 3 substituents selected from the group Doc: FP9907al.doc PS I 485/FP-9907(PCTtsagad/shtcorrectd pages of spedO03/0/O consisting offluoro and chloro groups: a CI C 4 alkyl group which has I to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of Ci C 2 alkyl. Ci C 4 alkoxy. fluoro and chloro group: and a phenyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of C C 2 alkyl. Ci C 4 alkoxy. fluoro and chloro groups.
A more preferable ether residue of ether derivatives is selected from the group consisting of a C 6
C
20 alkyl group: a Co 1 C:o alkenyl group having 1 to 3 double bonds; a C 3 Cs alkynyl group having one triple bond: a Ci C. alkyl group which has one substituent selected from the group consisting of C C 4 alkoxy. fluoro.
chloro and nitro groups: a Ci C 4 alkyl group which has 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of C C 2 alkyl. Ci C 4 alkoxv. fluoro and chloro groups: and a phenyl group which is optionally substituted with I to 3 substituents selected from the group consisting of CI C, alkyl, C 1 C4 alkoxy. fluoro and chloro groups.
A still more preferable ether residue of ether derivative is selected from the group consisting of a C 6
C
20 alkvl group: a CIo C 2 0 alkenyl group having 1 to 3 double bonds: a C 3 Cs alkynyl group having one triple bond: a Ci C 4 alkyl group which has one substituent selected from the group consisting of C C 4 alkoxy groups: and a Ci C 4 alkyl group which has 1 or 2 phenyl groups optionally substituted with 1 or 2 substituents selected from the group consisting of Ci C 2 alkyl. Ci C 4 alkoxv. fluoro and chloro groups.
A most preferable ether residue of ether derivatives is selected from the group consisting of a C C 20 alkvl group and a Clo Co alkenvl group having 1 to 3 double bonds.
An alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of "straight or branched chain Ci C, 1 alkyl group" such as methyl, ethyl. propyl.
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl. pentyl, isopentyl. 2-methylbutyl.
neopentyl, 1-ethylpropyl, hexyl. isohexyl, 4-methylpentyl, 3-methylpentyl. 2methylpentyl, 1-methylpentyl, 3.3-dimethylbutyl, 2,2-dimethylbutyl, 1.1dimethylbutyl, 1,2-dimethylbutyl. 1,3-dimethylbutyl, 2,3-dimethylbutyl. 2-ethylbutyl, heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, Doc: FP9907sl.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 1-32)/08.12.00 I -propylbutyl, 4,4-dimethylpentyl, octyl, I -methyiheptyl, 2-methylheptyl, 3methyiheptyl, 4-methyiheptyl, 5-methyiheptyl, 6-methylheptyl, 1 -propylpentyl, 2ethylhexyl, 5 ,5-dimethylhexyl, nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6methyloctyl, I1-propyihexyl, 2-ethyiheptyl, 6,6-dimethylbeptyl, decyl, 1-methylnonyl, 3methylnonyl, 8-methylnonyl, 3-ethyloctyl, 3 ,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8-dimethylnonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,1 1 -trimethyldodecyl, hexadecyl, 4,8,1 2-trimethyltnidecyl, I -methylpentadecyl, I 4-methylpentadecyl, 13,13 dimethyltetradecyl, heptadecyl, I 5-methyihexadecyl. octadecyl, I -methyiheptadecyl, nonadecyl, icosyl, 3,7,11,1 5-tetramethylhexadecyl and henicosyl groups; "straight or branched chain C 2
C
2 1 alkenyl or alkynyl group" such as ethenyl, I1propenyl, 2-propenyl, I -methyl-2-propenyl, 1 -methyl- I -propenyl, 2-methyl- I -propenyl, 2-methyl-2-propenyl, 2-ethyl-2-propenyl, I -butenyl, 2-butenyl, I -methyl-2-butenyl, I methyl- I -butenyl, 3-methyl-2-butenyl, I -ethyl-2-butenyl, 3-butenyl, 1 -methyi-3-butenyl, 2-methyl-3-butenyl, I -ethyl-3-butenyl, I -pentenyl, 2-pentenyl, I -methyl-2-pentenyi, 2methyl-2-pentenyl, 3-pentenyl, I -methyl-3-pentenyl, 2-methyl-3-pentenyl, 4-pentenyl, I rnethyl-4-pentenyl, 2-methyl-4-pentenyl, I -hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, hexenyl, cis-8-heptadecenyl, cis, cis-8,1 I -heptadecadienyl, cis, cis, cis-8, 11,14heptadecatrienyl, cis- I 0-nonadecenyl, cis- I 2-icosenyl, ethynyl, 2-propynyl, I -methyl-2propynyl, 2-methyl-2-propynyl, 2-ethyl-2-propynyl, 2-butynyl, I-methyl-2-butynyl, 2methyl-2-butynyl, I -ethyl-2-butynyl, 3-butynyl, I -methyl-3-butynyl, 2-methyl-3-butynyl, 1 -ethyl-3-butynyl, 2-pentynyl, I -methyl-2-pentynyl, 2-methyl-2-pentynyl, 3-pentynyl, I1- :0::.:methyl-3-pentynyl, 2-methyl-3-pentynyl, 4-pentynyl, I -methyl-4-pentynyl, 2-methyl-4pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl a nd "straight or branched chain C, C 21 alkyl group which has substituents selected from 00 the group consisting of alkoxy, halogen (hereinafter example fluorine, chlorine, bromine and iodine, preferably fluorine and chlorine) and nitro" such as methoxymethyl, :0 ethoxymethyl, methoxyethyl, ethoxyethyl, tifluoromethyl, trichioromethyl.
difluoromethyl, dichloromethyl, dibromomethyl, fluoromethyl, 2,2,2-trifluoroethyl, 2,2,2trichloroethyl, 2-bromoethyl, 2-chloroethyl, 2-fluoroethyl, 2-iodoethyl. 3 -chloropropyl, 4- 0.0. fluorobutyl, 6-iodohexyl, 2,2-dibromoethyl, nitromethyl, dinitromethyl, I -nitroethyl, 2nitroethyl and I ,2-dinitroethyl; and Doc: FP99O7a1 .doc P81485/FP-9907(PCT)/tsa/gadlshcorrected pages of spclO3/Ol/O1
"(C
6 Clo)aryl-(C, C 21 )alkyl group wherein said aryl moiety optionally has substituent selected from the group consisting of lower alkyl, lower alkoxy, halogen and nitro groups" such as benzyl, at-naphthylmethyl, j-naphthylmethyl, indenylmethyl, phenanthrenylmethyl, anthracenylmethyl, diphenylmethyl, triphenylmethyl, I -phenethyl, 2-phenethyl, I -naphthylethyl, 2-naphthylethyl, I -phenylpropyl, 2-phenylpropyl, 3phenylpropyl, I1-naphthylpropyl, 2-naphthylpropyl, 3-naphthylpropyl, I1-phenylbutyl, 2phenylbutyl, 3-phenylbutyl, 4-phenylbutyl, I1-naphthylbutyl, 2-naphthylbutyl, 3naphthylbutyl, 4-naphthylbutyl, I -phenylpentyl, 2-phenylpentyl, 3-phenylpentyl, 4phenylpentyl, 5-phenylpentyl, I -naphthylpentyl, 2-naphthylpentyl, 3-naphthylpentyl, 4naphthylpentyl, 5-naphthylpentyl, I -phenylhexyl, 2-phenylhexyl, 3-phenylhexyl, 4phenylhexyl, 5-phenylhexyl, 6-phenylhexyl, I -naphthylhexyl, 2-naphthylhexyl, 3naphthylhexyl, 4-naphthylhexyl, 5-naphthylhexyl and 6-naphthylhexyl.
A preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of a C, C21 alkyl group; a C 2 C21 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a C, C 2 1 alkyl group which has one or more substituents selected from the group consisting of lower alkoxy, halo and nitro groups; and a C, C2) alkyl group which has 1 to 3 C 6 CIO aryl groups which are optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups.
A more preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting ofa C, C21 alkyl group; a C 2 C21 alkenyl group having I to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group which has 1 to 4 substituents selected from the group consisting of C, C 4 alkoxy, halogen and nitro groups; and a C, C 6 alkyl group which has I to 3 C 6 C, aryl groups which are S* optionally substituted with I to 3 substituents selected from the group consisting of C,
C
4 alkyl, C, C 4 alkoxy, halo and nitro groups.
A more preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of a C, C21 alkyl group; a C 6
C
2 0 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a CI C 6 alkyl group which has one substituent selected from the group consisting of C, C 4 alkoxy and nitro groups; a CI C 6 alkyl group which has 1 to 3 substituents selected from the Doc: FP9907al.doc PS485/FP-9907(PCT)/tsa/gad/sh/corrected pages of spec/d03/01/01 group consisting of halo group: and a C 1
C
4 alkyl group which has 1 to 3 phenyl or naphthyl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of CI C 4 alkyl. Ci C 4 alkoxy. halo and nitro groups.
A more preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of a C 6
C
20 alkyl group: a Clo C 2 o alkenyl group having I to 3 double bonds: a C 3 C. alkynyl group having one triple bond: a Ci Ca alkyl group which has one substituent selected from the group consisting ofCi C 4 alkoxy and nitro groups; a Ci C 4 alkyl group which has 1 to 3 substituents selected from the group consisting of fluoro and chloro groups: and a C C4 alkyl group which has I to 3 phenyl groups which are optionally substituted with 1 to 2 substituents selected from the group consisting of Ci C 2 alkyl. CI C 4 alkoxy. fluoro and chloro groups.
A more preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of a C 6
C
0 alkyl group: a Clo C 2 o alkenyl group having 1 to 3 double bonds: a C 3
C
5 alkvnvl group having one triple bond: a C Ca alkyl group which has one substituent selected from the group consisting of C C 4 alkoxv, fluoro. chloro and nitro groups: and a Ci C 4 alkyl group which has 1 to 3 phenyl groups which are optionally substituted with 1 to 2 substituents selected from the group consisting of Ci C 2 alkyl. CI C 4 alkoxy. fluoro and chloro groups.
A still more preferable alkvl residue of N-alkylcarbamoyl derivative is selected from the group consisting of a C C 20 alkyl group: a Cio C 20 alkenyl group having 1 to 3 double bonds: a C 3 C; alkvnvl group having one triple bond: a Ci Ca alkyl group which has one substituent selected from the group consisting ofCi C 4 alkoxy groups: and a CI C 4 alkyl group which has 1 to 2 phenyl groups optionally substituted with 1 to 2 substituents selected from the group consisting of C C 2 alkyl.
Ci C 4 alkoxy. fluoro and chloro groups.
A most preferable alkyl residue of N-alkylcarbamoyl derivatives is selected from the group consisting of a C 6
C
0 alkyl group and a Co C 2 n alkenyl group having 1 to 3 double bonds.
In compound there are several functional groups to which the hydroxy protecting group, and the ester, ether and alkyl residues can be attached. Therefore a plurality of protecting groups and residues can independently exist by optional combination of these protecting groups and residues.
Doc FP9907sl.doc P81485/FP-9907(PCT)/tsa-gad-sh!English translation of spec (pages 1-321/08.12 00 23 A preferable pharmaceutically acceptable ester derivative of (Ia) is a derivative which has one or two of the ester residues at R 2
R
3 and/or R 5 A more preferable ester derivative is a derivative which has one or two of the ester residues at R 3 and/or A still more preferable ester derivative is a derivative which has one of the ester residues at R 3 or R 5 A most preferable ester derivative is a derivative which has one of the ester residue at R 3 A preferable pharmaceutically acceptable ether derivative of(la) is a derivative which has one or two of the ether residues at R 2
R
3 and/or A more preferable ether derivative is a derivative which has one or two of the ether residues at R' and/or R A still more preferable ether derivative is a derivative which has one of the ether residues at R' or A most preferable ether derivative is a derivative which has one of the ether residues at R 3 A preferable pharmaceutically acceptable N-alkylcarbamoyl derivative is a derivative having one of the alkvl residues.
The term "pharmaceutically acceptable salt" refers to a salt that is a useful medicament without significant toxicity.
Where compound and pharmaceutically acceptable ester, ether and N-alkyl derivatives of compound (la) have a basic group such as an amino group, these compounds can be convened into an acid addition salt by a conventional treatment with an acid. Such acid addition salts include inorganic acid salts such as hydrochloride. hydrobromide. sulfate and phosphate: organic acid salts such as acetate, benzoate. oxalate. maleate. fumarate. tartrate and citrate: and sulfonic acid salts such as methanesulfonate. benzenesulfonate and p-toluenesulfonate.
Where compound and pharmaceutically acceptable ester, ether and N-alkvl derivatives of compound (Ia) have an acidic group such as a carboxy group, these compounds can be converted into a base addition salt by a conventional treatment with a base. Such base addition salts include alkali metal salts such as sodium, potassium and lithium salts: alkaline earth metal salts such as calcium and magnesium salts; metal salts such as aluminium, iron. zinc. copper, nickel and cobalt salts: and quaternary ammonium salts such as ammonium salt.
When compound and pharmaceutically acceptable derivative of compound (Ia) are allowed to stand in the atmosphere, these compounds may take up water to form a Doc: FP9907sl.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 1-32V08.12.00 hydrate. The present invention includes such hydrates. Compound and pharmaceutically acceptable derivative of compound (Ia) may absorb a solvent to form a solvate. The present invention includes such solvates.
Compound and pharmaceutically acceptable derivative of compound (Ia) have several asymmetric carbons and therefore they can exist as several stereoisomers such as enantiomers and diastereomers in which each carbon has R or S configuration. The compound of the present invention encompasses individual enantiomers and diastereomers and mixtures of these stereoisomers in all proportions.
A preferable configuration of the compound of the present invention is shown below:
OH
O
CONH
2 HN 0 N NH HN N 0 0 0 N NH R1/ 00 R O O x Rd bH
OR
5 4 f R a 0 H CONH O RHN N 0 N NH O .O X R2aO' ZR 3 A preferable compound is selected from the following compounds: a compound wherein R 2 is a methyl group.
a compound wherein R 4 is a hydroxy group, a compound wherein X is a methylene group: or a compound wherein R 2
R
4 and X is selected in optional combination of(l), and for example: a compound wherein R 4 is a hydroxy group and X is a methylene group, and a compound wherein R 2 is a methyl group, R 4 is a hydroxy group and X is a methylene group.
Doc: FP9907sl.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 1-32)/08.12.00 A preferable compound of formula (la) is selected from the following compounds: a compound (1a) wherein the protecting group for a hydroxy group is selected from the group consisting of "tetrahydropyranyl or tetrahydrothioipyranyl group".
"~silyvi group", "aralkx'l group". "aralkylo xycarbonyl group". "Il-(aliphatic acyloxy)- (lower alkyl.) group". -(cyclI oalkyl carbony]loxy)-(lower alkyl) group". I lower alkoxycarbonvloxv)-(lower alkyl) group". "I -(cy-cloalkylIoxycarbonylIoxy-)-( low\er alkyl) group". "phthalidNvl" and 'oxodioxolenvimethyl group".
(ii) a compound (la) wvherein the protecting group for a hydroxy group is selected from the group consisting of tetrahydropyran-2-yl. 4-methoxv-tetrahvd ropv-ran-4-\vl.
tetrahydrothiopyran-2-yl. trimethylsilyl. triethylsil tert-butyldimethylsilyl. di( tentbutvl )methylsilyl. diphenylimethvislivl. benzyl. diphenvlmrethy'l. triphenlmethyl. 4methvlbenzvl. 4-methoxvbenzvL. 2-itrobenizvl. 4-nitrobenzvl. 4-chlorobenz\-l.
benzyloxy'carbonvl, 4-methoxybenzyloxycarbonyl. 2-nitrobenzy'loxyc arbonyl. 4nitrobenzyloxvcarbonvl. acetoxymethyl. propiony lox ymethy 1. butyr\ loxymethyl.
pivaloyloxymethyl, valervloxymethvl. 1 -acetoxvethyl. butvrvloxvethvl. I pivalovloxvethyl. cvclopent\vlcarbon\yloxv-methvl.. cvlclohex\vlcarbonvloxvlmeth\vl. I cyc lopentvlcarbonvloxyethyl. I -cy-clohexvlicarbon\vloxveth\vl.
methoxycarbonvloxymethyl. ethoxycarbony'loxymethyl. propoxycarbonyl oxv methyl.
isopropoxvcarbonyloxymethvl. butox ycarbonyl oxym ethyl.
isobutoxycarbonyloxymethvl. I -(methoxvcarbonv loxv)ethvl. I (etho x carbon vl oxv)ethvl1. I isopropoxycarbonvl oxvetvl1.
c\vclopentvloxycarbonvloxvm ethylI. c\vclohex\vloxv-carbon\vlox\vmeth\vl. I (cvclopentvloxv,,carbon\vlox\ )ethylI. I-(c\-clohex-l oxv.carbonv-loxv)eth\vl. phthalidvl.
(5-phenvl-2-oxo-l1.'3-dioxolen-4-vl I)meth-,l. [5-(4-methvlphen\'l )-2-oxo- I.3)-dioxolen- 4-vl]methvl, (5-methyl-2-oxo- .3-dioxolen-4-vl )methvl and (5-ethvl-2-oxo- I .3dioxolen-4-yl)methyl group.
(111) a compound (la) wherein the protecting group of hydroxy group is selected from the group consisting of trimethylsilyl. tert-butyldimethylsily'l. tri phenv'imethyl.
benzyL. 4-methoxybenzyl. acetoxymethyl. propionyloxymethyl. butyry'loxv methyl.
pivaloyloxymethyl, valervloxymethyl. cyclopentylcarbonvloxymethyl.
cyclohexylcarbonyloxymethyl. methoxycarbonyloxymethyl.
ethoxycarbonyloxymethyl, propoxycarbonvloxymethyl, isopropoxycarbonyloxymethyl, butoxycarbonyloxymethyl, isobutoxycarbonyloxymethyl, cyclopentyloxycarbonyloxymethyl, Doc: FP9907slI.doc PSI 485/FP-9907(PCTl/tsa-gad-shlEnelish translation of spec (pages 1-32)/08.12.00 cyclohexyloxycarbonyloxymethyl, (5-phenyl-2-oxo-1,3-dioxolen-4-yl)methyl, methylphenyl)-2-oxo-1,3-dioxolen-4-yl]methyl, (5-methyl-2-oxo- ,3-dioxolen-4yl)methyl and (5-ethyl-2-oxo-l,3-dioxolen-4-yl)methyl group.
A preferable ester derivative of compound (Ia) is selected from the following compounds: (iv) an ester derivative of compound (la) wherein the ester residue is R 6 CO- or R 6
OCO-
group in which R 6 is selected from the group consisting of hydrogen; a Ci C 2 1 alkyl group; a C 2
C
2 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a Ci C 2 alkyl group substituted with 1 to 4 substituents selected from the group consisting of lower alkoxy, halo and nitro groups; a Ci C21 alkyl group substituted with I to 3 C 6 Clo aryl groups which are optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups; and a C 6 Co0 aryl group which is optionally substituted with 1 to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups.
an ester derivative of compound (la) wherein the ester residue is R 6 CO- or RO6COgroup in which R 6 is selected from the group consisting of hydrogen; a CI C 2 alkyl group; a C 2
C
2 1 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group substituted with 1 to 4 substituents selected from the group consisting ofC, C 4 alkoxy, halo and nitro groups; a C, C 6 alkyl group substituted with 1 to 3 C 6
CI
0 aryl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting ofC, C 4 alkyl, C, C 4 alkoxy, halo and nitro groups; and a C 6 CIo aryl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of Ci C 4 alkyl, C, C 4 alkoxy, halo and nitro groups.
(vi) an ester derivative of compound (la) wherein the ester residue is R 6 CO- or R 6
OCO-
group in which R 6 is selected from the group consisting ofa C -C 2 alkyl group; a C 6
C
20 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group substituted with one substituent selected from the group consisting of CI C 4 alkoxy and nitro groups; a Ci C 6 alkyl group substituted with I to 3 substituents selected from the group consisting of halogen; a CI C 4 alkyl group substituted with 1 to 3 phenyl or naphthyl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of C, Doc: FP9907al.doc PS1485/FP-9907(PCTtssa/g ad/sh/corrected pages of spec/l3/01101
C
4 alkyl. C 1
C
4 alkoxy, halo and nitro groups: and a phenyl or naphthyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of C, C 4 alkyl. C, C 4 alkoxy. halo and nitro groups.
(vii) an ester derivative of compound (la) wherein the ester residue is R 6 CO- or
R
6 OCO- group in which R 6 is selected from the group consisting of C, C2 0 alkvl group; a Clo C 20 alkenyl group having 1 to 3 double bonds: a C 3 C. alkynyl group having one triple bond: a Ci C 4 alkyl group substituted with one substituent selected from the group consisting of C C 4 alkoxy. and nitro groups: a Ci C 4 alkvl group substituted with 1 to 3 substituents selected from the group consisting of fluoro and chloro groups: a Ci C 4 alkyl group substituted with 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of C C2 alkyl. Ci C 4 alkoxy, fluoro. and chloro groups: and a phenyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of C
C
2 alkyl. Ci C 4 alkoxy. fluoro and chloro groups.
(viii) an ester derivative of compound (Ia) wherein the ester residue is R"CO- or
R
6 OCO- group in which R 6 is selected from the group consisting ofa C, C 20 alkvl group: a C 0 o C 20 alkenyl group having 1 to 3 double bonds: a C 3
C
5 alkynyl group having one triple bond: a Ci C 4 alkyl group substituted with one substituent selected from the group consisting of C C 4 alkoxv. fluoro. chloro and nitro groups: a Ci C 4 alkyl group substituted with 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of Ci C 2 alkvl. C, C 4 alkoxy. fluoro. and chloro groups: and a phenyl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of Ci C 2 alkvl. C, C 4 alkoxy. fluoro and chloro groups.
(ix) an ester derivative of compound (la) wherein the ester residue is R"CO- or
R
6 OCO- group in which R 6 is selected from the group consisting of a Ct C 2 o alkyl group; a Clo C 20 alkenyl group having 1 to 3 double bonds: a C 3 Cs alkvnvl group having one triple bond; a Ci C 4 alkyl group substituted with one substituent selected from the group consisting of C C 4 alkoxy groups; and a C C 4 alkyl group substituted with 1 to 2 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting ofCi C 2 alkyl. C, C 4 alkoxy. fluoro and chloro groups.
Doc: FP9907sl.doc P81485/FP-9907(PCTI/tsa-gad-sh/English iranslation of spec (pages 1-32)/08.12 00 an ester derivative of compound (la) wherein the ester residue is R 6 CO- or R 6
OCO-
group in which R 6 is selected from the group consisting of a C 6
C
20 alkyl group; and a Cio C 20 alkenyl group having 1 to 3 double bonds.
A preferable ether derivative of compound (la) is selected from following compounds: (xi) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a C, C21 alkyl group; a C 2
C
2 1 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a C, C2 alkyl group which has 1 to 3 substituents selected from the group consisting of lower alkoxy, halo and nitro groups; a C, C 2 1 alkyl group which has 1 to 3 C 6 CIo aryl groups which are optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups; and a C 6 Clo aryl group which is optionally substituted with I to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups.
(xii) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a Ci C21 alkyl group; a C 2 C21 alkenyl group having I to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a C, C 6 alkyl group which has 1 to 4 substituents selected from the group consisting of C, C 4 alkoxy, halo and nitro group; a C, C 6 alkyl group which has 1 to 3 C 6
C
0 aryl groups which is optionally substituted with I to 3 substituents selected from the group consisting of C C 4 alkyl, Ci
C
4 alkoxy, halo and nitro groups; and a C 6 Ci, aryl group which are optionally substituted with I to 3 substituents selected from the group consisting of C C 4 alkyl, C,
C
4 alkoxy, halo and nitro groups.
(xiii) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a CI C 2 1 alkyl group; C 6
C
20 alkenyl group having 1 to 3 double bonds; a C 2
C
6 alkynyl group having one triple bond; a Ci C 6 alkyl group S* which has one substituent selected from the group consisting of C, C 4 alkoxy and nitro groups; C, C 6 alkyl group which has 1 to 3 substituents selected from the group consisting of halo group; a C, C 4 alkyl group which has 1 to 3 phenyl or naphthyl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of C, C 4 alkyl, C, C 4 alkoxy, halogen and nitro groups; and a phenyl or naphthyl group which is optionally substituted with I to 3 substituents Doc: FP9907al.doc PS1485/FP-9907(PCT)/tsa/gad/sh/corrected pages of spec/03/01/01 selected from the group consisting of C C 4 alkyl. Ci C 4 alkoxy. halo and nitro groups.
(xiv) an ether derivative of compound (Ia) wherein the ether residue is selected from the group consisting of a C 6
C
20 alkyl group: a Co 1
C
20 alkenvl group having 1 to 3 double bonds; a C 3 Cs alkynyl group having one triple bond: a Ci C 4 alkyl group which has one substituent selected from the group consisting of C C 4 alkoxv and nitro groups; a Ci C4 alkyl group which has 1 to 3 substituents selected from the group consisting of fluoro and chloro groups: a Ci C 4 alkyl group which has 1 to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting ofCi C: alkvl. C 1
C
4 alkoxy. fluoro and chloro groups: and a phenyl group which is optionally substituted with I to 3 substituents selected from the group consisting ofCI C 2 alkyl. Ci C 4 alkoxy. fluoro and chloro groups.
(xv) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a C 6 Co 2 alkyl group: a CIo C 20 alkenyl group having 1 to 3 double bonds; a C 3 C. alkynyl group having one triple bond: a Ci C 4 alkvl group which has one substituent selected from the group consisting of C C. alkoxv.
fluoro. chloro. and nitro groups: a Ci C 4 alkyl group which has I to 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting of CI C 2 alkyl. Ci C 4 alkoxy. fluoro and chloro groups: and a phenvl group which is optionally substituted with 1 to 3 substituents selected from the group consisting of Ci C 2 alkyl. C1 C 4 alkoxy. fluoro and chloro groups.
(xvi) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a C 6
C
20 alkyl group: a CIo C 20 alkenyl group having 1 to 3 double bonds; a C 3 C? alkynyl group having one triple bond: a Ci C 4 alkyl group which has one substituent selected from the group consisting of Ci C 4 alkoxy group; and a Ci C 4 alkyl group which has Vor 2 phenyl groups optionally substituted with 1 or 2 substituents selected from the group consisting ofCi C 2 alkvl. C C alkoxy.
fluoro and chloro groups.
(xvii) an ether derivative of compound (la) wherein the ether residue is selected from the group consisting of a C 6
C
20 alkyl group and a Clo C( alkenvl group having 1 to 3 double bonds.
A preferable N-alkylcarbamoyl derivative of compound (la) is selected from the following compounds: Doc: FP9907sl.doc P81485/FP-9907(PCTI/tsa-gad-sh/English translation of spec (pages 1-32)/08.12.00 (xviii) an N-alkylcarbamoyl derivative of compound (la) wherein the alkyl residue of the N-alkylcarbamoyl derivative is selected from the group consisting of a C, Cz 2 alkyl group; a C 2 C21 alkenyl or alkynyl group having 1 to 3 double or triple bonds; a Ci C 2 alkyl group which has 1 to 4 substituents selected from the group consisting of lower alkoxy, halo and nitro groups; and a Ci C21 alkyl group which has 1 to 3 C6 CI0 aryl groups which are optionally substituted with 1 to 4 substituents selected from the group consisting of lower alkyl, lower alkoxy, halo and nitro groups.
(xix) an N-alkylcarbamoyl derivative of compound (la) wherein the alkyl residue is selected from the group consisting of a Ci C21 alkyl group; a C2 C21 alkenyl group having 1 to 3 double bonds; a C2 C 6 alkynyl group having one triple bond; a Ci C 6 alkyl group which has 1 to 4 substituents selected from the group consisting of C, C 4 alkoxy, halo and nitro groups; and a CI C6 alkyl group which has I to 3 C6 Clo aryl groups which are optionally substituted with 1 to 3 substituents selected from the group consisting of C, C4 alkyl, Ci C4 alkoxy, halo and nitro group.
(xx) an N-alkylcarbamoyl derivative of compound (la) wherein the alkyl residue is selected from the group consisting of a CI C 21 alkyl group; a C6 C20 alkenyl group having 1 to 3 double bonds; a C2 C6 alkynyl group having one triple bond; a C, C6 alkyl group which has one substituent selected from the group consisting of C, C4 alkoxy and nitro groups; a C, C6 alkyl group which has I to 3 substituents selected from the group consisting of halo groups; and a Ci C4 alkyl group which has I to 3 phenyl or naphthyl groups which are optionally substituted with I to 3 substituents selected from the group consisting of C C4 alkyl, C, C4 alkoxy, halo and nitro groups.
(xxi) an N-alkylcarbamoyl derivative of compound (la) wherein the alkyl residue is selected from the group consisting of a C6 C20 alkyl group; a Clo C20 alkenyl group having 1 to 3 double bonds; a C3 C5 alkynyl group having one triple bond; a CI C4 alkyl group which has one substituent selected from the group consisting of C, C4 alkoxy and nitro groups; a C, C4 alkyl group which has I to 3 substituents selected from the group consisting of fluoro and chloro groups; and a CI C4 alkyl group which has 1 to 0* 3 phenyl groups which are optionally substituted with 1 or 2 substituents selected from the group consisting ofCi C2 alkyl, Ci C4 alkoxy, fluoro and chloro groups.
(xxii) an N-alkylcarbamoyl derivative of compound (la) wherein the alkyl residue is selected from the group consisting of a C6 C20 alkyl group; a Cio C20 alkenyl group Doc: FP9907al.doc P81 485/FP-9907(PCTgsa/gad/sh/coectcd pages of spec03/01/I0 having 1 to 3 double bonds; a C3 C 5 alkynyl group having one triple bond; a Ci C 4 alkyl group which has one substituent selected from the group consisting of C C 4 alkoxy, fluoro, chloro and nitro groups: and a CI C 4 alkyl group which has 1 to 3 phenyl groups which are optionally substituted with I or 2 substituents selected from the group consisting of C C- alkvl. Ci C 4 alkoxy, fluoro and chloro groups.
(xxiii) an N-alkylcarbamoyl derivative of compound (Ia) wherein the alkyl residue is selected from. the group consisting of C 6
C
20 alkyl group: a C 0 o C 2 o alkenyl group having 1 to 3 double bonds: a C3 C 5 alkynyl group having one triple bond: a Ci C 4 alkyl group which has one substituent selected from the group consisting of C C 4 alkoxy groups: and Ci C 4 alkvl group which has I or 2 phenyl groups optionally substituted with 1 or 2 substituents selected from the group consisting of C C, alkvl.
Ci C 4 alkoxy. fluoro and chloro groups.
(xxiv) an N-alkylcarbamoyl derivative of compound (la) wherein the alkvl residue is selected from the group consisting of a C Co 0 alkyl group and a Clo Co alkenvl group having 1 to 3 double bonds.
A more preferable compound (la) is selected from group to (iii): group (iv) to group (xi) to (xvii): group (xviii) to (xxiv) in optional combination of these groups. for example: (xxv) a compound (la) wherein the protecting group for a hydroxy group is and the ester residue is (iv).
(xxvi) a compound (la) wherein the protecting group for a hydroxy group is (ii) and the ester residue is (xxvii) a compound (Ia) wherein the protecting group for a hydroxy group is (iii) and the ester residue is (vi).
(xxviii) a compound (Ia) wherein the protecting group for a hydroxy group is and the ether residue is (xi).
(xxix) a compound (la) wherein the protecting group for a hydroxy group is (ii) and the ester residue is (xii).
(xxx) a compound (Ia) wherein the protecting group for a hydroxy group is (iii) and the ether residue is (xiii).
(xxxi) a compound (Ia) wherein the protecting group for a hydroxy group is and the alkyl residue is (xviii).
Doc: FP9907sl.doc P81485/FP-9907(PCT .'tsa-gad-sh/English translation of spec (pages 1-32)/08.12 00 32 (xxxii) a compound (Ia) wherein the protecting group for a hydroxy group is (ii) and the alkyl residue is (xix).
(xxxiii) a compound (Ia) wherein the protecting group for a hydroxy group is (iii) and the alkyl residue is (xx).
The following Tables 1 and 2 are intended to illustrate typical compounds and (la) of the present invention and are not intended to limit the scope of this invention.
Doc FP9907sl.doc P81485/FP-9907(PCT/ltsa-gad-sh/English translation of spec (pages 1-32 V08.12.00 Table 1 0:R 5a R 4 R3 a'
HN
4' (Ib) Exemp. X RR1 R d Comp.
No.
II1 Me MeCHH OH H 2 CH-, Me H HOH H .3CHl MIe Me H H H 4 1CH, Me %,Ie 1A7 IOH H 'CH, M e Me !A8 OH H 6 'CH, Me IMe A9 OH H 7 1 CH,1 Me Me A10 OH H 8 H, me IMe A12 OH H 9 Me Me ;A14 OH H 1CH,~ Me Me A15 OH H ii CH-b Me IMe A 16 OH H 12 CH, Me MIe 'A17 OH H 13CH-, Me Me A18 OH H F14 ICH-, Me Me A20 OHl H CH,, Me Me A22 OHl H [16 1-CH 2 Me M e OLE OHl H 17 C12 Me IMe E iOH H 18 CH, Me Me LEN OHl H CH2 1 Me Me 1 CDS 'OH H 21 CR 2 Me Me DPP OHl H 22CR, Me Me TMPP OH H Doc: FP9907s2.doc Doc:FP907s2doc P81485/FP-9907(PCTI/tsa-gead-shlEnglish translation of spec (pages 33-U5: Tables 1&2)/08.12.00 23 I-2 e Me 1 NPP O 24 CH, Me Me MPP OH H 26 CH, Me Me ND OH H 26 ;CH, 1 Me Me KN OH IH 28 Me Me TMP OH H 129 Me M CeI MeIPA OHH 'CH, Me IMe BZA OH H F31 CH, Me Me NBZ OH H 32CH, Me 'Me iCB OH H 33C- 2 MIe M-e 'MB OH H i 4CH, Me Me 'EB OH H CH, Me iMe MO O H 36 CH, Me IMe ID FOH H 37 1 CH, M e Me MDD OH H 38 MH ve M4e MTD OH H -39 CH 2 Me MIe M~HD OH *H 1 CH, Me Me DM0 OH H 141 CH, Me Me DMD OH iH .42 CH, Me Me DMDD OH 1H 43 CH, Me Me DMTD OH 'H 144 CH-, Me Me *DH HH CHI H H H OH 'H 46 CHI H Me A7 OH 'H 47 CHI H IMe 'A8 1OH H 48 CH 2 H Me ,A9 OH 'H 49 CH-, H Me AlO OH H
C-
2 H Me A12 OH 'H 51 CHI H Me A14 OH H 52 jCH, TH Me A15 'OH 'H 53 CHI H Me A16 OH H 54CH-, IH Me A17 OH 'H Doc. FP9907s2.doc Doc.FP907s2doc P1485/FP-9907(PCT/sa-gad-shEnglish translation of spec (pages 33-U5 Tables 1&2)/08 1200 58 Cl-, H MVIe OH H 59 Cl-, H Me LE OH H JCH- H Me 'LEN OH H 61 ICH, H Me CES OH H 62 ClHI H Me :CDS OH H :63 CH-, H M"e DP OH H 64 CHI H Me TMPP OH H CHI H Me ,NPP OH H 66 CH, H M e :MPP OH H 67 1 CH,~ H Me CP TOH H 68- CH-, H MIe ND 'OH H 169 ICH, H Me TCN OH H CH, H Me MP OH H 71 CH 2 H Me CPA OH H 72 FCH, H Me BZ 1OH H 73 ICHI H Me NBZ OHH 74 CH-, H Nie CB OH H CHI~ Me MB OHH 76 HMIe .EB OH H 77 Cl-, H Me MOD OH H ICH, H Me 'MD OH H 879 CH 2 H Me 'MDD OH- :H
{CH
2 HMe DMD OH H 81) CH 2 H Me DMD OH H 84 CH, ~H Me DMDD JOH H
CH
2 H I Me fDMTD OH H 86 CH 2 H Me DMHD OH H Doc FP9907s2.doc Doc: P99O~~doc P81485/FP-9907(PCT)/sa-gad-shFneish translafion of spec (pages 33-85: Tables U-2u08.12.00 CH,~ Me Me H OH A17 96 CH7 .Me Me H TOH A 18 97 CH, Me Me H OH 'A2, 98 Me Me H OH2 99 ,CH, Me Me HOH OLE 100 CH, Me Me .H OH LE 101l CH, Me Me H OH LEN 102 CH, ,Me Me H OH CES 103) CH-, Me Me 1H OH iCDS 104 CHI 'Me Me H OH DPP 105 CH-I {Me Me- H OH 'TMPP 106 ICH-, Me 1 Me ~H OH NPP 107 CH- Me Me ,H iOH MPP 108 IC- Me Me H OH CP 109 CH, Me .MeHOHN 110 NCI Me .Me H OH TCN III1 CH,1 Me M e H OH MP 1112 CH, Me :Me H OH CPA 113 Cl-, Me Me H OH BZ 114 CH, Me Me HOH NBZ 115 Cl- 2 Me IMe H OH CB 116 Cl-I Me Me H OH MB 117 CH 2 Me Me H OH IEB 118 Cl- 2 Me Me H OH jMO Doc. FP9907s2.doc Doc P99~s2doc P81485/FP-9907(PCT)/sa-gad-shEnglish translation of spec (pages 33-85 Tables 1&21/08 12.00 122 !CH, Me Me H OH MHD 123 i CH, Me IMe H iOH DM0I ~124 CH-, Me Me H OH DMNJD I 125 CH, jMe Me 'H OH DMCD I 126 CH, Me Me H OH :DM\TD 12-7 ,CH, Me 1Me H 1OH 'DMHD 12 8 CHl H M ke H iOH A 7 12 CH, H Me H OH A8 130 CH, ,H Mve H 'OH ,A9 131 ICH, H 'e H OH AID *12 CH, H Me H IOH A 12 '13 CH,' H Me H OH A14 134 CH, H Me IH OH 135 CH, H M ve H 'OH 'A16 136 lCH, 'H Me H OH 1 137 H MNIe H 'OH 1A18 138 CH-I H Me ,H IOH 139 ICH, H Mve H OH A22 140 JCH- :H Me H OH OLE 141 ,H Me H OH LhE 142 ICH-, 'H Me H OH LEN 143 CH, H Me H OH CES 144 CH, H Me H HCDS 145 l-I 2 H Me H OHDP 146 CA 2 ~H Me H OH TPP 147 CH2 H Mve H OH NPP 14 ____HMe___HOHNP 148 CH, H Me HOH MPP 149 CH 2 'H [Me HOH CP 150 CH, HM OH jND Doc: FP9907s2.doc Doc FP90s2.oc P81485/FP-99O7(PCT/tsa-2ad-shEnplish translation of spec (pages 33-8U Tables 1&2)/08 1200 159 '159 Cl- H Mvie HOHM i I OH I mo 160 CH,~ H MIe H O H MvID 161 CH-, HHOH ND 161 H Me H OH MHD 165 CH, Me H OH MDI 163 H Me H 'OH DMD 167 CHI H OMH, DM0D 165 CH, [H Me H 'OH DM4D H7 MH, M ,Ie AH DMDD 171__ 1_ KeN 9H H_ 1662 CH, e 1 173 CHI HI Me H O2' H *DT 174 Me e_ A_4_H H 176 CHI He Me H1 H DHD 169 CH-, Me M e A17 H H 170 CH, Me Me A18 H H 1710 CH, Me M e A22 H IH 11 ICH, Me Me AO H H 174 CH 2 Me Me LE4 H H Doc FP9907s2.doc Doc:FP9O~s2doc P8I485/FP-99O7(PCT/tsa-g2ad-shEnglish translation of spec (pages 33-85 Tables 1I&2)/08.12.00 183) CHi Me Me LEN H H 184 CH, Me Me CES H H 185 CH-, Me Me CDS H H 186 CH, me Me DPP H H 187 CHI~ Me Me ITMPP HH S18 CH-, Me IMe NPP H Hi 189 Cl-, Me Me MPP 'H H 190 ClHI Me Me CP H H 191I CH, Me e ND H 192 CHI Me Me TCN H H 193 CH2 Me Me .1 P H H 194 CH, Me Me CPA H 'H 195 CH, Me Me BZ H *H 196 CH, Me Me NBZ H 'H 197 CH, Me Me i CB H H 198__ CH-_ MeMH_ 198 Me Me MB H H 199 CH, Mie 1Me EB }H H i200 M Me MD tHH 202 CH,1 Me Me MDD 'H 'H 123 CH-, Me 'Me MTD H H 204 Cl- 2 Me Me IMHD H H CH2 Me Me DM0 H H 26 CH, 'Me Me DMDD H H 207 CH, Me Me iDMDD H H 208 CH, Me Me .DMTD H H 209 CH-, Me Me DHD H A8 22 CH 2 Me Me 'H H A9 211 CH2 Me Me HH A8O 212 CH 2 jMe Me H H 12 Doc. FP9907s2.doc Doc. P99072.doc P8 1485/FP-9907i PCT)/tsa-gad-shlEnghIsh translation of spec (pages 33-85 Tables 1 &2)/08.!12.00 21H-CH i Me *Me Hj H IA14 r 1- C H 2 fme Me H HA1 21 H e H JH A16 217 JCH 2 Me Me H H A17 218 'CHI Me Me H H A18 219 CH-, Me Me H H A2 0 22 CHI Me Me H H 2 221 1CH, Me Me H iH OLE 222) CH-, Me Me 'H iH LE 224 'Cl-I Me Me H H LEN CH, M I '2247 CH, Me Me H H LEDN CH, Me 'Me H H CESP CH-_ me_ MeHH I 2260 Mke Me H H :CDS CH., Me Me H H DPP 28 CHI Me Me H iH NDP 2293 CH2 Me Me .H H TN 230 CHI I Me Me H H 'MPP 235 CH, IMe Me H ,H ICPA __36 CH Me-, eHH 232 CH, Me Me H H NDZ 23 8 CH, IMe Me H H TCB I H -M 240 [CH, Me Me iH H MP 235 ~CH2 Me Me H H CPA 236 CH, Me Me H H MZ 23 CH2 Me Me H 'H INBZ 239 CH, Me Me H H MBD 241 CH 2 Me Me H H MOD 242 CH, Me 'Me H H DM Doc: FP9907s2.doc Doc:FP907s2doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 33-85 Tables 1&2)/08. 12.00 252 C- 2 Me Me A8 A A08 A8 253 CH,- I Me (Me H iAO() A9 :254 !CH, Me Me H 1AGIO M 2-55 CH, Me Me ,H AO,1-' A12 256 CH-, MIe Me *H jA014 A 14 257 CH, Me Me 'H A015 .2 58 'CH, Me Me H fA0l6 ;A16 9 CH-, IMe Me H A017 A17 260 iCH, Me Me H jA018 A 16 1 .26] CH, Me Me H OLEO OLE .262 CH-, Me Me H LE 26 JCH-, Me Me H LENO LEN 264 iCH, Me Me H CESO E CMMe e S 1
CES
.265 CH-) Me IMe H TCDSO Fp 26 CHCDSM PP P 266 Me Me H DPPO DPP 270 CH2 Me Me H TMCPO TPP 268 71 CH, Me Me H NPPO NPP 269 CH., Me Me H 'MPPO MPPC 1270) CH, Me Me H CPO MP CH Me__Me______CPA 275 CH, Me Me H BDO ND 272 CH2 'Me Me H NO TN 275 CHI Me (Me H CBO CB 278 CH 2 Me Me H MBO JMB Doc FP9907s2.doc Doc: P99072.doc P81 485/FP-9907{ PCT)/tsa-ead-shJEnglish translation of spec (pages 33-85. Tables 1 &2 /08.12.00 287 CH, iH Me H A016 A 16 288 CH 2 H Mie H A017 A17 -289 jC-I i H Me H A018 18 ii 290 iCH-, H M ie H I OLEO OLE .291 CHi H Me 'H LEO LE 292 CH, 'H Me H LENO LEN 293 CH- H Me H CESO CES 294 CH 2 H Me H CDSO CDS 295 CH, 2 H Me ,H 1 DPPO DPP 296 CH., H Me H TMPPO JTMPP 297 CH-, H Me H NPPO NPP ,:298 CH- H Me H MPPO MPP 299 CH- H Me ,H CPO CP 300 CH, H Me 'H NDO ND '301 CH, H Me H [TCNO TCN '302 CH 2 H Me H MPO NIP 303 CH- H Me H CPAO CPA 304 CH, 1H %,Me H BZO CH, H i Me 'H NBZ() :NBZ 306 CH- H Me H CBO CB -307 CH, H Me H IMBO i MB .308 CH- H Me 1H EBO 'EB 309 CH, Me Me A7 OH A7 310 CH 2 Me Me A8 OH A8 Doc FP9907s2.doc P8]485IFP-9907(PCT,'tsa-ead-shEng 5 sh translation of spec (pages 33-85 Tables 1 12.00 311 Cl-I 2 Me A9 OH A9 CH, Me A OH i A9 31 2 CHI Me Me lA OH AlO 133 CH, Me 'Me A12 OH A12 3'1 H e Me A14 OHA14 315 CH-, Me Me AJ5 OH 316 CH MMe A16 OH A 1317 'CH, I Me Me A17 OH A17 318 'CH, Me Me A18 OH IA18 '319 CH, Me Me 'OLE O0H 'OLE '320 iCH, Me Me 'LE OH1- LE CH, LEN O 13] Me Me OHLEN 32 CH,~ Me M'e CES OH CES 3-23) CH, Me Me CDS O0H CDS 324 CH, Me Me DPP IOH DPP 325 1CH, Me Me iTMPP OH TMPP 3_ 2 6 1 Mee___OHP .326 CH, Me MIe NPP OH NPP 327 CH, M Ne Me ND MP OH MPD 332 C- 'Me Me CPA OHCPA 32CH, Me M e ND OH ND .)330 CH-, Me Me TN OH TN 3.31 5 CH-, Me Me MPB OH CBP 1332 Cl-, Me Me CPA OH CPA 333 CH, Me Me EB 'OH B 340 H Me NB OH NA9 CH Me 337 CH 2 Ae10 E OH EB1 342 ~CH- 2 H ]Me A12 OH A12 Doc: FP9907s2.doc Doc: P99072.doc P1I4851FP-9907 PCTh/tsa-gad-shIEn~ish translation of spec (pages 33-85 Tables 1 &2 i/O8.12.00 349 CHI H Me LE OH LE 350 iCH, 'H Me LEN IOH LE-N 351 CHI H Me 1CES OH CES 352 CH, *H Me ;CDS OH CDS :353 CH, i H Me DPP OH DPP 354 CH, H Me TMPP OH iTMlP P 355 1C- 2 H Me INPP OH NPP 356 Cl- HMe MP jOH MIP 357 CH, ,H M e ICP OH :CP 358 H M.Ie ND OH ND CH-, H IMe TCN OHTC 360 CH, H 1Me MP OHMP 361 CH, H Me CPA OH CPA 3 62 CH,1 H Me BZ TOH BZ 363 CH, H M4e NBZ OH NBZ 364 CH) ,H .Me CB OH CB 365 CH, 'H Mvle M B OH MB 36 6 CH 2 H %,Ie EB OH EB 37 Cl-I Me Me A7 H A7 136 8 1CH- Me Me A8 H A8 36 9 jCH, Me Me A9 'H A9 370 CH, Me Me AID H iAIO 371 CH, Me FMe 'A12 'HA1 372 C-, 2 Me Me A14 H A14 .373 CHb Me [Me A15 H 374 CH, JMe Me A16 HA1 Doci FP9907s2.doc Doc FP90s2.oc P81485/FP-9907(PCT)/tsa-gad-shlEnglish translation of spec (pages 33-85 Tables &2)/08 12.00 382 Me Me DPP H *DPP 383 ICH-, Ne Me TMPP H TNIPP 384 CH 2 M1,1e M4e 'NPP H P 385 CH, Me iMe NIPP H KIPP 386 CH-, Me Me I CP H CP 387 CH, NIe Me ND H ND 388 CH-, Me Me TCN H TCN 389 CH, MIvIe %,Ie %IP H NI P .390 CH, MIe Me 'CPA H CPA 391 iCH, M e .MVIe BZ H BZ 392 jCH,1 Me Me NBZ H NBZ 33 CH, [Me .Me CB H CB 394 CH-, M Me MB H MB 39 I e MIe EB 'H iEB -396 S Me Me H 1OH H Me e9A OH H .398 S Me Me A8 OH *H 3)99 S Me M e A9 OH [400 S Me M e .A10 OHI 401 S Me Me A12 OH H 402 S Me Me A14 'OH H 4 0 3 Me iMe A15 OH H 404 S jMe Me A16 OH~ H 405~~I __MMA7_H_ 406 S Me 'Me A18 OH H Doc- FP9907s2.doc Doc P997s2doc P8l485/FP-9907(PCT/sa-eadshEngij 5 h translation of spec (pages 33-85 Tables 1200 4 12 S Me iMe CES OH *H 413 S NIe Me :CDS IOH H 414 S Me Me IDPP OH H 415 iS Me IMe TMPP OH H '416 S Me Me ,NPP OH H 417 S Me Me MPP OH H 418 S Me Me CP OH H 419 S Me 1Me ND OH H 420 S Me Me TCN OH 'H 421 S Me Me MP OH H 422 S Me Me CPA OH H 423 Me e BZOHq 424 S Me Me BZ OHH 424 S Me IMe CBZ OH H *426 SMe iMe MB OH H 42 7 M Ie Me EB 1OH H 428 5 M\4e MO OH H 429 5 M Ie Me MD OH H .430 {S M le 'Me 'MDD OH H 431 S Me -Me iMTD OH H 432 5 'MI Me MHD OH :H 433 S Me 'Me DM0 OH 'H 434 S Me 'Me DMD 1 OH H ~4 35 s M Me DMDD OH H 436 S Me Me DMTD OH H 437 5 Me Me DMHD jOH H 438 S Me Me H OHf A7 Doc: FP9907s2.doe Doc:FP907s2doc P81485[P-997PCT/sagad-sh/Enizsh translation of spec (pages 33-85: Tables 1l&2)/08 12.00 4 39 S Me Me H OH A8 440 S jMe Me H OH A9 441 ~S 'Me Me H OHA1 442___eMe_ OH__ JA12 443 S Me Me H 'OH A1I4 44 S IMe Me H 445 is Me Me 'H IOH A 16 446 SMe 'Me H OH A]7 447 IS Me Me H OH A18 448 SMeMe H OH 449 S M e Nie H OH A22 450 S Mle %-1e H OH 'OLE 451 S, Me Me H OH LE 452 S Me Me H OH LEN 453 S M'Ve %.Ie H 'OH CES 454 'S Me IMe H 1OH CDS 455 5 %,Ie Me 'H iOH DPP 456 'S Me IMe iH OH TMPP 457 S Me Mle 'H OH NPP '458 5 Me IMe H OH MPP 459 S Me Me H OH CP 460 S Me kle H OH ND .461 'S \Me M\Ie H IOH TCN 462 'S iiMe Me H IOH NI1P 1463 S Me Me .H OH CPA 464 S Me INIe H OH 'BZ 465 5 Me M e H OH NBZ 466 S Me Me H OH CB 467 5 Me Me H OH M B 468 S M e Me iH OH LB 469 S Me Me jH OH MO 47 5 Me 1 Me jHOH MD Doc FP9907s2.doc Doc' P99072.doc P8]485/FP-9907(PCT)/tsa-gad-shEnglis, translation of spec (pages 33-85 Tables 1 &2)/08.12.00 471 S Me Me H OH MDD 472 S Me Me H OH MTD 473 S Me 1Me H OH IMHD 474 S e Me H OH 475 S MMe H OH DMD 476 Me Me H OH DMDD 477 s Me Me H OH DMTD 47 Me me 1HOH JDMHD 479 S Me iMe H A07 A7 480 1 Me Me H As ;.A8 481 Me Me H A09 A9 i482 S Me Me H AO01 I 483 vS Me Me H AO1 A12 484 S Me Me ,H A014 A14 :485 S Me .Me H AQIS 486 'S IMe \,,IMe H A016 A16 487 !S M,,Ie 'Me H A017 A17 :488 S I Me Me H A018 A18 489 S I Me Me H OLEO OLE 490 1 e .Me H LEO LE 491 S Me Me H LENO LEN 1492 IS .Me H CESO ICES 493 S Me I H iCDSO CDS [494 S Me Me H DPPO DPP 495 S Me Me H TMPPO TMPP 496 S 1 Me Me H NPPO 'NPP 497 S Me 'Me H MPPO MPP 498 5 Me Me H 1 CPO CP 499 S Me Me H NDO ND 500 Me Me H TCNO TCN 501 S Me Me H MPO IMP 502 5 Me Me H CPAO CPA Doc: FP9907s2.doc P81485/FP-9907( PCT)tsa-ead-sh/English translation of spec (pages 33-85. Tables 1 &2)/08.12.00 510 S :Me Me A9 OH A9 511 S Me Me A0 OH [12 S 'Me Me A12 0OH :A12 513 S Me Me A14 OH A14 1514 S Me Me A15 OH 51 I e_ IM__eA16__OHA_6 516 S iMe Me A17 OH A17 516 S IMe Me A18 OH A8 518 S Me Me A1 'LE OH OLE 518 5 Me Me LE OH LE 520 S Me Me 'LEN OH LEN 521 S Me IMe CES OH CES 22 5 Me Me CDS OH CDS 523 5 Me Me DPP iOH DPP 524 S I Me .Me TMPP OH TMPP 52 S e_ OH 5256 Me Me INPP OH NPP 527 S 'Me Me, CP OH CP 528 S Me Me ND OH ND 529 5 'Me 1Me TCN OH TCN 30 S Me Me MP OH MP 531 S Me Me CPA OH CPA 32 S Me Me iBZ 1OH 'BZ S33 S M Ie NBZ OH NBZ 34 S Me jMe jCB OH CB Doc: FF9907s2.doc Doc:FP9O~s2doc P8]485/FP-99O7(PCT)/sa-gad-sh/En 2 -ijsh translation of spec (pages 33-85 Tables 1&2/08.12.00
MB
EB
IOH
OH
'MB
EB
Me Me !C60C OH H Me 'Me !C70C OH 'H 1539 jCH, Me Me C80C 'OH 540 CH, Me i Me C90C OH H 541 CH2 Me iMe CI0OC 'OH H 542 'CH, 'Me MIe CIIOC OH H i 543 iCH, Me i Me C12OC i OH H -544 CH, Me Me M10OH 544 CH., Me Me MMA2IO2 OH H 545 CH-, i Me Me MN'14 OH H .547 CH, Me Me DMAIO OH H 548 CH,1' Me Me DMA 12 OH H 549 CH- Me Me DMA4 OH H 550 {CH, I Ie Me H OH 1551 JCH, Me Me H OH 552 1 CH2 Me Me 'H 1 OH I C8OC 553 CH, Me Me H Hi C9OC 554 CH, Me Me H OH C100C Me CH, e 1Me H I OH iCIIOC 556 1 CH, I e Me H OH CI2OC 557 CH 2 Me Me H OH i MMAIO 558 CH, Me Me 'H OH MIMA12 559 CH 2 Me 'Me H OH ;IMA14 560 CH Me Me 'H 'OH DMAIO
CH
2 IMe 'Me 'OH DMA12 i CHI Me
OH
563 CH 2 H 'Me C60C OH 564 CH 2 H Me C7OC OH 565 CH 2 H Me C80C OH 566 CH 2 H Me C90C OH Doc FP9907s2.doc P81485/FP-9907 PCT)/tsa-gad-sh/Enish translation of spec (pages 33-85 Tables 1 &2)/08,1200 Doc: FP99O7s2.doc P81485IFP-99O7(PCT)/tsa-gad-sh/English translation of spec (pages 33-85: Tables 1&2)/08. 12.00 Me Me CVj 599 CH, I e M 1 OH H 600 CH 2 Me Me C16 OH H 601 [CH Me Me C6 OH 1A7 '602 CH-I 2 Me Me i C6 jOH 1603 jCHI iMe Me 'C6 OH A9 604 fCH-{ Me Me C6 OH M 605 jCH 2 Me Me :C6 OH AK 606 CH, Me Me C6 'OH A14 607 CH, Me Me C6 OH AI1 608 CHI Me Me C6 OH A16 .609 CHI Me Me ;C6 OH .A17 .610 CH Me Me 1 C6 OH AI8 1 611 CHI Me 1 Me C6 OH OLE I .r 4- :612 iCH 2 Me Me C6 OH LE 613 CH Me IMe C6 OH LEN M64C Ie .C6 OH C 0 614 CHI i Me I MeeC6 OH C70C i 616 CHI Me Me C6 OH COC 616 CHI Me Me C6 OH .618 !CHI Me MN't e C6 OH 1c~oc i 6 1 H, M h e 16 OH 618 CH2 Me Me6 'OH 619 0 CHI Me .Me C6 OH CIoC .620 CH-I Me Me C6 OH CIOC 621 CH 2 Me Me C6 OH MMA12 S622 CH~ Me .Mh~e :C6 MMACI2C 623 CH2 Me Me C6 1 OH :MMAI4 624 CHI Me Me C6 OH DNAIO 625 CI jMe Me iC6 OH DMA12 626 CI Me 'Me 1C6 lOR DMAI4 627 CHI Me Me CS jON A7 628 CHI Me Me IC8 OH A8 629 CI 2 Me Me C8 OH A9 630 CH 2 Me Me C8 OH Doc- FP9907s2.doc P8I485/FP-997(PCT)/tsa-ad-shEnelish translation of spec (pages 33-85 Tables 1&1/08.12 00 639 ;CH, Me Me 'C8 1OH LEN 640 iCH-, Me Me C8 OH 641 CH-, MIe M e iC8 OH *642 CH, 'Me Me C8 iOH 63 CH, Me Me C8 OH CQOC 644 CH, Me Me .C8 OH iCIOOC 645 CH, Me 'Me C8 OH CI 646 CH-, Me Me C8 OH C120C 647 CH-, Me IMe C8 OH MMAIO i68 CH-, Me Me C8 OHMA1 648 I, 'M eO 'MNA14 .649 CH, i Me M e C 8 OH IMMAI4 ,651 ICH, Me Me C8 OH DMA12 651 jCH, Me Me I8 OH DMA14 ICH, Me Me CIO OH A4 654 CH, Me Me CIO 'OH ,A8 654 CH, Me Me CIO OH A9 656 1 CH, Me Me CIO OH :A9O 656 CH-, Me Me C 10 OH A12 1658 CH, Me Me CIO ~OH A14 CH-, Me Me CIO OH A16 660 CH 2 IMe Me CIO OHA1 662 CH-, Me Me CIO OH 1A18 Doc FP9907s2.doc Doc P99~s2doc P8l485/FP-9907(PCT)/tsa-gadsh/Englsh translation of spec (pages 33-85 Tables 1&2)/08.12 00 663 CH 2 Me Me CIO oH OLE 664 CH 2 Me Me CIO OH LE 665 Cl] 2 Me Me CIO OH LEN 666 CH 2 Me Me CIO OH 667 CH 2 Me Me CIO OH 668 CH 2 Me Me CIO OH 669 Cl] 2 Me Me CIO OH 670 CH 2 Me Me CIO OH CIOOC 671 CH 2 Me Me CIO OH CI 1OC 672 Cl] 2 Me Me CIO OH C120C 673 Cl] 2 Me Me CIO OH MMAIO 674 CH 2 Me Me CIO OH MMA12 675 CH 2 Me Me CIO OH MMAI4 676 Cl] 2 Me Me CIO OH DMAIO 677 CH 2 Me Me CIO OH DMA12 678 CH 2 Me Me CIO OH DMA14 679 Cl] 2 Me Me C 12 OH A7 680 CH 2 Me Me C12 OH A8 681 Cl] 2 Me Me C12 OH A9 682 CH 2 Me Me C12 OH 683 CH 2 Me Me C12 OH A12 684 CH 2 Me Me C12 OH A14 685 Cl] 2 Me Me C12 OH- 686 Cl] 2 Me Me C12 OH A16 687 Cl] 2 Me Me C 12 OH A17 688 Cl] 2 Me Me C12 oH A18 689 CH 2 Me Me C12 OH OLE 690 Cl] 2 Me Me C12 OH LE 691 Cl] 2 Me Me C 12 OH LEN 692 CH 2 Me Me. C12 OH 693 CH 2 Me Me C12 OH 694 CH 2 Me Me C12 OH Doc: FF9907a].doc Doc: P99O~I~doc P148 5/FP-9907(PCT)/sa/gad/sh/corrected pages of spcc/O3IOI/OI 700 CH-, Me Me 'C12 OH M A I 701 CH, i Me Me C12 OH NMI 702 CH, M e .Me C I OH 703 CH, N-Me NIe C12 OH D.%IA I 2 704 Me M'le C12 OH D.NI.AI4 705 CHI H N1e C5 OH H :706 CH, 'H Mie C6 OH H 707 CH, H N.Me C7 'OH 'H 708 CH, H NIe C 8 OH H .709 CH, H NMIe C9 OH H 710 'CH, H Me CIO OH H 711 [CH, H Me C I 1 OH H '712 CH, H N-Me C12 iOH 1H 713 1 CH, Me :C13 OH 'H 714 CH-, H 'Me 'C14 'OHH 715 iCH-, H Me CI5 OH H- 716 .H Me C16 OH H 717 {CH, H Nie 'C6 OH A7 718 CH- 2 H Me 'C6 'OH A8 '719 CH, 'H *M C6 OH A Q 720 CH,~ 1H Me C6 OH AIO 721 CHI) H Me :C6 OH Al1- 722 CH, H 'Me jC6 OH A14 723 CH, H Me C6 OH 724 Cl- 2 H Me C6 OH A16 725 CH 2 H Me C6 OH A17 726 CH-) H Me C6 OH A18 Doc: FP9907s2.doc Doc:FP907s2doc P81485FP-9907(PCT/1sa-gad-shIEnglish translation of spec (pages 33-85 Tables 1&2 1/08.12.00 734 ICH, H 1 Me I C6 OH CIOOC 73 5 iCH, ,H Me iC6 OH CIIOC .736 lCH, H Me C6 OH C120C 737 CH, H I Me C6 'OH NIAI1 738 CH, H i Me C6 OH 'MMAI2 739 CH 'H -Me C6 O0H PyMMAI14 '740 CH, H Me C6 I OH DAIO .741 CH, H Me C6 OH DMA 1 742 ,CH, 'H Me .C6 OH 'DMA 14 743 CH, H Me C8 OH A7 744 CR2, H Me C8 OH A8 745 CH, I H Me C8 'OH iA9 '746 ,CH- H Me C8 OH 747 lCH, H \,Ie C8 OH iA12 1748 iCH, I N-1e I C8 I OH A1 .748 CH 'i H Me C8 OH 750 -CH H e C8 OH A16 1751 CH, H Me C8 1 OH A17 752 CH_ H Me OH A18 750 CR 2 H Me C8 OH OLE 754 CRH H Me C8 'OH :LE 752 CH- H Me C8 OH LEN 756 CH, H Me C8 OH 757 CH, H Me C8 OH 755 CR 2 H Me C8 OH Doc. FP9907s2.doc P81485/FP-9907 PCT)/tsa-ad-shlEnglish translation of spec (pages 33-85 Tables 1 &2)/08 12.00 764 CH-I H Me (78 OH IM N1A 12 765 1 CH, H Me C8 '0H- MMA 14 766 CH-, H Me '8OH D MA 767 CH., H Me (78 OH DIN A 12 768 CH,-1 H Me (78 OH !DM\IA 14 769 CH, 'H Me (710 OH A 7 770 C7H, H Me CIO OH A8 771 H Ne C(71 OH A 9 772 H Ie (710 OH M 773 CH-, H M e C(71 OH AI2 774 i CH, H \/Ie (710 OH A 14 775 CH, H Me (710 OH 776 C7H, H M NIe (710O OH A16 777 C7H, H Me (710 OH A17 778 IA, H 'Me (710 OH A18 779 C7H- H i %4e (710 OH OLE '780 CIA,~ H Me (710 'OH ILE .781 C7H-, H (710 OH LEN 782 C7H, H NMe (7I0 OH (760(7 783 (71-2 H Me (710 OH (770(7 784 C7H, H Me (710 OH (780(7 785 (7H,1 H Me (710 OH (790(7 786 7H 2 IH Me (710 'OH (7100( 787 CR 2 H 'Me (710 OH (7110(7 [7 88 C H 2 H Me (710 OH (7120(7 789 CHI H Me jCIO OH IMMAIO 790 (7-2 ~H 1 Me (710 OH jMMA12 Doc: FP9907s2.doc Doc P99~s2doc P148SIFP-9907(PCT/tsaEad-shIEnish translation of spec (pages 33-85. Tables 1 &2)/08 12.00 799 CHI H1Me C12 IOH A2 800 CH-, H IMe C12 OH A14 801 CH, H Me c 12 OH 802 CH, H Me C12 'OH A16 803 CH, *H %,Ie C12 OH A17 .804 CH, *H Me .C12 [OH A18 .805 1CHb H Me C12 'OH iOLE 806 'CH, H Me C02 OH LE 807 CH-, H IMe '012 [OH LEN 808 CH, 'H Me C12 OH C0 809 CH, H IMe C 12 OH .810 C- 2 H Me c 2 OH 811 C 2 H 0e OH 812 CH,1 H M e ,C12 OH CIOOC 813) CH, H %,Ie :C12 OH i cuo1c 814 CH) H %1Me C12 OH 'CI2OC 815 CH, H Me C12 OH NMAIO 816 CH, H Nie :C12 OH MMAl2 817 CH, H Me C 12 OH M MA14 818 CH-, H ;IMe iC12 'OH DMAIO 819 CH-, H Me iC 12 OH DMA12 1820 CIA 2 H Me 1C12 OH DMA14 Doc FP9907s2.doc Doc.FP9O~sdoc P81485/FP-9907(PCT)/sa-gad-shEnglish translation of spec (pages 33-85 Tables 1 &2)/W812.00 Table 2
OR
/R
-NH
"0 ,N NH (1k)
CH
3 O' OF Exemp. R I RI]I R) R Comp.I No. j_ 891 Me IMe HH 892 Me Me A7H 893 Me Me A8 H 894 jMe 'Mle I A9 H 895 Me jMe AIO H 896 Me M'e !A12 H 897 Me Me :A14H 898 Me Me 899 Me Me :A16 iH 900 1 M Me A17 H 901 Me Me A18 H 902 Me {Me 1C60C H ~903 Me Me C70C H 904 Me Me C80C H 905 Me Me C90C H 906 Me jMe C1OOC H 907 Me Me C1 1OC H 908 Me Me C120C H 909 Me Me MMAIO H 910 Me Me DMAIO H Doc: FP9907s2.doc Doc FP9Os2.oc P814851FP-9(907(PCT/sa-gad-sh/English translation of spec (pages 33-85 Tables 1 &2)/08.12 00 :911 Me 'Me C5 H M eM CI 912) Me Me C7 H 913 Me Me iC8 H 915 Me TMe C9 H 916 Me Me CI0 H 917 Me CII H 918 !Me Me 012 H 919 Me Me C03 ,H 920 Me Mve C14 H ~921 Me N'e H .A6 922 Me NIe H A 7 .923 Me Mle H A8 *924 'Me Me H A9 925 Me Mle H 926 Me Me H All 927 Me Me H A 12 928 Me Me H jA13 929 [Me Mle H A14 930 [Me Mle H 931 me Me H A16 932, 'Me %,Ie :H A17 93 Me N14e H A18 934 Me Mie H 935 Me 1Me H C0 936 Me Me iH 937 Me Me H C9OC 938 Me M e 'H CIGOC 939q Me Me 'H CI 1OC 940 Me Me H C120C 941 Me C2 H H 942 Me C2 A7H Doc: FP9907s2.doc Doc:FP907s2doc P81 485/FP-9907 PCT~itsa-gad-shlEnlish translation of spec (pages 33-85: Tables 1 &2 /08l 12 00 943 Me C2 A8 H [944me C2 A9 H [945 IMe C? AIO H 946 Me C2 A1 H 947 me C2 A14 (H 948 me C2 A15 :H 949 me C2 jA16H 950 Me /C2 !A17 H 951 (Me A18 H 952 Me C2 (C60C H 953 (Me C2 C70C H 954 1Me C2 'C80C H 955 Me C-'C0 H .956 Me C2 CIOOCH 957 me C2) CI 1OC H- 958 M~e 1C2 I12OC H Ie 1C2 MMA1O
H
960 Me C2 DMAIO H Me C5 961 Me C2 C6 H 964 Me C2 C8 963 Me C2 C7 'H 964 TMe jG2 CIO H 967 Me C2_ I_ I_ 968 Me C2 C912 [966 Me C2 13 Hl- 970 Me JC2 CI14 H [968 Me C2 C? 972 Me C2 H A7 9 73 Me C2) H A8 974 Me C2 H A9 Doc FP9907s2.doc Doc:~~P IP9~~o 485/FP-9907(PCT)/tsa-gad-shEnlish translation of spec (pages 33-85 Tables 1l&2)/08.12.00 Doc: FP99O7s2.doc P81485/FP-99O7(PCT)/tsa-gad-shEnglish translation ofspec (pages 33-8U Tables i&2)/08.12.OO Doc: FP99O7s2.doc P8I485IFP-99O7(PCT /sa-ead-shIEnglish translation of spec (pages 33-8U Tables l&2)/08 12.00 1039 me H ClIOC 1040 ]Me C 1 H C120C 1041 Me C6 H H [1042 Me C6 A7 H 43 Me C6 !A8 H 1044 Me C6 A9 H 1045 Me C6 lAIO H 1046 Me C6 1A12 *H ~1047 Me C6 A 14 H 1048 [Me C6 '1049 [Me C6 A16 H 1050 Me C6 A17__ H 1050 Me C6 A18 H 1051 Me C6 C60C H 1052 Me jC6 C70C H 1053 Me JC6 C80C H 1054 Me C6 C90C iH 1056 Me C6 C9O0C H 1056 Me C6 Tciooc [H 1057 Me C6 C120C H 1058 Me C6 MI2O H 11059 Me C6 IAIO H 101 eC6 110602 Me CC6H 1061 Me C6 C7 H 1064 Me C6 IC8 H- 1063 Me C6 C9 H 1064 Me 1 C6 CIO H 1067 Me C6 ICI H 1066 Me C6 C12 H 1069 Me C6 C13H 1070 Me C6 C14H Doc: FP9907s2.doc Doc:FP9O~s2doc P81485/FP-9907(PCT)/tsa-gad-shEnglish translation of spec (pages 33-85 Tables Il&2)/08 12.00 1071 1Me C6 H A6 1072 Me C6 H !A7 1073 e C6 H A8 1074 Me 1C6 H .A,9 1075 Me )C6 H AlO 1076 Me C6 H A]lI 1077 me 1C6 H A 12 1078 Me C6 H A 13 1079 Me C6 H A 14 1080 IMe C6 H A 1081 Me C6 'H .A16 1082 'me C6 H A 17 1083) Me C6 H 'A18 1084 Me IjC6 H 1085f e 6 C OCI 1086 Me !C6 H 1087 Me }C6 H 1088 Me C6 H 10898 Me TC6 H CIIOC 1089 Me 1C6 H 1091 Me C 12 H H 1092 Me IC12 A7 H 1093) Me Il I A8 H 1094 Me 'C1I A9 H 1095 Me C12 AIOH 1096 Me C12 1097 Me 1 C12 A14 H 1098 Me C12 A15 H 1099 Me C12 A16 H 1100 Me C12 'A17 H 110 C12__A18__H 1101 Me C12 C60C H Doc: FP9907s2.doc Doc:FP907s2doc P8148S/FP-9907(PCT/tsa-gad-shEnplish translation of spec (pages 33-8i Tables 1&2)/08 12.00 Doc: FP99O7s2.doc P81I4851FP-99O7(PCT)/tsa-ead-shlEnglish translation of spec (pages 33-85. Tables 1 &2 )/08.1I2.00 Doc: FP99O7s2.doc P81485/FP.9907(PCT)/tsa-gad-shIEngish translation of spec (pages 33-U5 Tables U&2/08. 12.00 1174 iH Me H AO 1175 H Me iH 1176 1H Me H A] I .1177 H Ni'e H 1A12 '1178 H Me H A I 1179 iH Me 'H A 14 1180 H M Ie H 1181 TH 'N Ie H A16 1182 H Me H A17 1183) H Mle H A 18 1184 H M e IH 1185 H Me 'H 1186 H Mie 'H 1187 1H Mie H 1188 H Nie ,H C I OC 1189 H \.Ie H C1 1190 H Me H CI 2OC 1191 H
HH
1192 TH A7 H 11931 T H C2 A8 H 1194 H C2 A9 H 11195 Hj C2) A10 H 1196 H C2 A12 H 1197 H C ]A14 H 1198 H jC2 JA15 H Doc: FP9907s2.doc Doc F99O~~doc P8 1485/FP-9907i PCT)/sa-g-ad-shlEngjjsh translation of spec (pages 33-8; Tables 1&2),'08.12 00 1199 'H C2 A16 H 1200 HC2 A17 H 1201 H C? A18 H 1202 }H C2 C60C H 1204 H C2 tC80C H 1205 jH C2 C90C H 120 H C2 CIOOC H 1207 H C2 ICIIOC H 1208 H C2 C 120C H 1209 jH C2MNIA1O H 1210 H C2 DNIAIO H 11211 IH C2 C5 H [1212 HC2 C6 H 1213) H C2 !C7 H 11214 H C2 C8 H 1215 H C2 C9 H 1216 H C2 FCIO 'H __217 H_ C2 1217 H C2 CI2 H 1219 H 7C- C13 H 129H 1C2 C14 H 1220 H {C2 H1 A6 1221 H C2 H A7 1222) H 1 C2 H 8A 1223 H C2 H A9 1224 H CH A9O 1226 H C2 H A 11 1226 H C2 H All 1227 H C2 H 1228 H C2 H A14 123 0 H C2 H Doc: FP9907s2.doc Doc:FP907s2doc P81485/FP-9907(PCT)/tsa-gad-shlEnelish translation of spec (pages 33-85 Tables 1&2/08 12.00 Doc: FP99O7s2.doc P8I48S/FP-99O7(PCT)/tsa-2ad-sh/En 2 lish translation ofspec (pages 33-85 Tables 1&2)/08 12.00 Doc: FP99O7s2.doc P81 4851FP-9907( PCT)/tsa-gad-shlEnglIish translation of spec (pages 33-85 Tables 1 &2)(/08.12-00 1295 'H C6 AO H 1296 H C6 A12 H 1297 H C6 A14 H 1298 H C6 A15 H 1299 H C6 A1l6 H 13)00 H 6A17 'H 1301 H C6 A18 'H ff102 H C6 IC60C 'H 13031 H C6 C70C H 1304 H C6 C80C H 1305 H C6 C90C H 1306 H C6 CIOOC H 1307 'H C6 ~C 11C H '1308 H C6 'CI20C *H 13 09 1H C6 MIMA 10 H 1310 H C6 DMAIO H 131H C6 iC5 H [1312 H C6 C6 H 131 H C6 C7 H 13)14 H C6 C8 H [13115 H C6 C9 1316 fH C6 CIO H 1317 H jC6 C H 1318 H C6 12 H 1319 H C6 13 H K320 H C6 C14 H 132 H C6 H A6 132H C6 'H A7 1323 H C6 H A9 1324 H C6 H 1326 H C6 H All Doc: FP9907s2.doc Doc. P99072.doc P8 1485/FP-9907( PCT)/rsa-gad-silEnp-lish translation of spec (pages 33-85. Tables 1 &2 /08.12.00 1327 'H C6 H A12 1328 H C6 H A 13 1329 IH C6 H A14 1330 JH C6 [H 1331 H C 6 H A16 32H C6 HA1 1333 n H C 6 H 1A18 1 334 H C6 J 1335 H C6 JH 136HC6 H 11'in)7 H C6 H 1C90C 1338 H C6 H ,40HC6 H Cl 2OC 341 [H 1342 H C 1)A 13441 1 IA 135HC12 HI H 1346 H C12 A12 H 1343 tH C12 A148 1.344 H C12 iA95 H 1345 IH C12 A16 H 1346 H C12 A17 H 1347 H C12 1A18 H 1348 H C12 C6C 1349) H C12 C70C H S1350 H C12 C80C H 1352 H C 12 TC90C H 1356 H C12 ClOOC H 1357 H C 12 ClIOC H 1358 H C12 C 120C H Doc: FP9907s2.dGc Doc:FP9O~s2doc P81485/FP-9907(PCT)/sa-gad-sh/Eng-lish translation of spec (pages 33-85. Tables 1&2)/08.12.00 Doc: FP99O7s2.doc P81 4851FP-9907( PCT)/tsa-gad-shlEnglish translation of spec (pages 33-85 Tables 1 &2 )/08.1I2.00 1391 Me C4 H H 1392 Me C5 H H 1393 Me C7 H H 1394 Me C8 H H 1395 Me C9 H H 1396 Me CIO H H 1397 Me CII H H 1398 Me C13 H H 1399 Me C14 H H 1400 Me CI5 H H 1401 Me C16 H H 1402 H C4 H H 1403 H C5 H H 1404 H C7 H H 1405 H C8 H H 1406 H C9 H H 1407 H CIO H H 1408 H C11 H H 1409 H C 13 H H 1410 H C14 H H 1411 H C15 H H 1412 H C16 H H
V
V V V. *V V V V V V V
V
V
V.
V
In Tables I and 2 Exemp. comp. No. is exemplification comnpound number,
CH
2 is methylene group, Me is methyl group, OH is hydroxy group, A6 is hexanoyl group, A7 is heptanoyl group, A8 is octanoyl group, A9 is nonanoyl group, Doc: FP9907al.doc Doc: P99O~I~docPS I485/FP-9907(PCT)Ixsa/gad/sh/corrected pages of spec/O3IOIIOI is decanoyl group, A]12 is lauroyl group, A 14 is myristoyl group, A] 15 is pentadecanoyl group, A] 6 is palmitoyl group, A] 17 Is heptadecanoyl group, Al 8 is stearoyl group, is arachidoyl group, A22 is behenoyl group, A07 is heptanoyloxy group, A08 is octanoyloxy group, A09 is nonanoyloxy group, AOl 10 is decanoyloxy group, AOl 12 is lauroyloxy group, AG 14 is myristoyloxy group, AO 15 is pentadecanoyloxy group, A016 is palmitoyloxy group, AO017 is heptadecanoyloxy group, AOl 18 is stearoyloxy group, y: *A020 is arachidoyloxy group, A022 is behenoyloxy group, OLE is olcoyl group, LE is Iinoleoyl group, :0:LEN is linolenoyl group, CES is cis-I 1-cicosenoyl group, CDS is cis-lI 3-docosenoyl group, DPP is 3,3-diphenyipropionyl group, *00* TMPP is 3-(3 ,4,5-trimethoxyphenyl)propionyI group, NPP is 2-(4-nitrophenyl)propionyl group, MPP is 3-(4-methylphenyl)propionyi group, CP is 3-chioropropionyl group, 0* ND is I 2-nitrodecanoyl group, TCN is trans-cinnarnoyl group, MP is 3-methoxypropionyl group, 130c: FP9907al.doc Doc: P99O~I~doc P I485/FP-9907(PCT)/tsa/gad/Sh/corrced pages of speciO3/OIIOI 77 CPA is 4-chlorophenylacelyl group, BZ is benzoyl group, NBZ is nitrobenzoyl group, CB is 3-chlorobenzoyl group, MB is 2-methoxybenzoyl group, EB Is 4-ethylbcnlzoyl group, OLEO is olcoyloxy group, LEO is Iinoleoyloxy group, LENO is Ilinolenoyloxy group, CESO is cis- I I -eicosenoyloxy group, CDSO is cis- I 3-docosenoyloxy group, DPPO is 3,3-diphenyipropionyloxy group, TMPPO is 3-(3,4,5-trimetboxyphenyl)propionyloxy group, NPPO is 2-(4-nitropbenyl)propionyloxy group, MPPO is 3-(4-methylphenyl)propionyloxy group, CPO is 3-chioropropionyloxy group, NDO is I 2-nitrodecanoyloxy group, TCNO is trans-cinnamoyloxy group, MPG is 3-methoxypropionyloxy group, CPAO is 4-chiorophenylacetyloxy group, BZO is benzoyloxy group, :NBZO is nitrobenzoyloxy group, CBO is 3-chlorobenzoyloxy group, MBO is 2-methoxybenzoyloxy group, EBO is 4-ethylbenzoyloxy group, MO is 2-methyloctanoyl group, MD is 2-methyldecanoyl group, MDD is 2-methyldodecanoyl group, MTD is 2-methyltetradecanoyl group, MHD is 2-methyihexadecanoyl group, DM0 is 2,2-dimethyloctanoyl group, is22dmtydea lgop DMDD is 2,2-dimethydecanoyl group, DMTD is 2,2-dimethyltetradecanoyl group, Doc: FP9907a].doc Doe: P99O~~doc P1485IFP-9907(PCT)/tsa/gad/shcorrcted pages of spcJO3IOIIOI DM4HD is 2.2-dimethvlbexadecanoyi group.
C2 is ethyl group.
C0 is propvl group.
C4 is butyl group.
is penty] group.
C6 is hexvl group.
C7 is heptyl group.
C8 is octyl group.
C9 is nonvI group.
is decyl group.
C IlIis undecyl group.
C12 is dodecyl group.
C I 'Iis tridecyl group.
C 14 is tetradecyl g-roup.
C 15 is pentadecyl group.
C 16 is hexadecyl group.
is hexyloxvcarbonyl group.
is heptylox ycarbon vI group.
is oct\vlox\-carbon\vl group.
is nonvioxycarbonyl group.
ClIOOC is decyloxycarbonyl goup.
ClII OC is undecvloxvcarbkon\ I grOLIp.
C 1 20C is dodec%-loxvcarbo--n% I group.
is 2-methyl decanovI group.
NMN4A1I2 is 2-methvldodecano\ l group.
MMNIA 14 is 2-methvhtetradecanov I croup.
DMA 10 is 2.2-dimethyldecanoyl group.
UM A 12 is 2.2-dimethvldodecanovl crourn.
D.MA 14 is 2.2-dimethvltetradecano\ I group.
Doc FP9907s2.doc Doe F9907s.doc P81 485/Fp-g(riPCT 'lsa-gad-shiEngl1ish translation of spec (pages 33-85i Tables 1 &2/08 12.00 In a compound of formula (Ib): the compound wherein R' is a methyl group. R' is a methyl group. R 3 is a hydrogen atom. R 4 a is a hydroxy group. is a hydrogen atom and X is a methylene group represents A-500359A (exemplification compound No. 1): the compound wherein R' is a methyl group. R' is a hydrogen atom. is a hydrogen atom. R 4 a is a hydroxy group. is a hydrogen atom and X is a methylene group represents A-500359C (exemplification compound No. 2): the compound wherein R is a methyl group. R is a methyl group. R' is a hydrogen atom. is a hydrogen atom. R' is a hydrogen atom and X is a methylene group represents A-500359D (exemplification compound No. 3): the compound wherein R' is a hydrogen atom. R- is a hydrogen atom. is a hydrogen atom. is a hydroxy group. is a hydrogen atom and X is a methvlene group represents A-500359G (exemplification compound No. 45): and the compound wherein RI is a methyl group. R" is a methyl group. is a hydrogen atom. R-a is a hydroxy group. is a hydrogen atom and X is a sulfur atom represents A-500359M-2 (exemplification compound No. 396).
In Tables 1 and 2: preferable compounds include compounds of ex.emplification compound No.
(exemp. comp. No.) 1 to 254. 280 to 283. 309 to 312. 338 to 341. 367 to 370. 396 to 482. 508 to 513. 537 to 588. 502 to 704. 78 to 820. 891 to 910. 914 to 990. 1091 to 1160. 1164 to 1210. 1214 to 1240.. 1341 to 1300. 1394 to 1401 and 1405 to 1412: more preferable compounds include compounds of exemplification compound No.
Sto 3. 7 to 11.45. 49 to 53. Q0 to 94. 131 to 135. 172 to 176. 213 to 217. 396. 400 to 404. 537 to 543. 550 to 556. 563 to 569. 576 to 582. 592 to 600. 708 to 716. 891 to 908. 921 to 940. 1091 to 1108. 1121 to 1158. 1171 to 1190. 1341 to 1358 and 1371 to 1390: most preferable compounds include compounds of exemplification compound No.
1 to 3. 7 to 11.45. 49 to 53. 90 to 94. 131 to 135. 537 to 543. 550 to 556. 563 to 569, 576 to 582. 594. 710. 891. 895. 925. 1091. 1141. 1145. 1175 and 1341: Doc FP9907s2.doc P81485/FP-9907(PCT Itsa-gad-sh/English translation of spec (pages 33-85. Tables I&2 /08 12.00 that is exemp.comp.No.1 represents the compound wvherein R' is a methyl group. R' is a methyl group. R'a is a hydrogen atom. is a hydroxy group. R'a is a hydrogen atom and X is a methylene group: exemp.comp.No.2 represents the compound w.herein R is a methyl group. R is a hydrogen atom. R'a is a hydrogen atom. R, is a hvdroxv group. Ra is a hydrogen atom and X is a methylene group: exemp.comp.No.3 represents the compound wherein R' is a methyl group. R is a methyl group. is a hydrogen atom. R a is a hydrogen atom. is a hvdrogen atom and X is a methylene group: exemp.comp.No.7 represents the compound \\herein R' is a methyl group. R is a methyl group. is a decanoyl group. is a hydroxy group. is a hydrogen atom and X is a methylene group: exemp.comp.No.8 represents the compound w\herein R is a methyl group. R is a methyl group. R3a is a lauroyl group. is a hydroxy group. R is a hydrogen atom and X is a methylene group: exemp.comp.No.9 represents the compound wherein R' is a methyl group. R is a methyl group. Ra is a myristol group. R4, is a hydroxy group. is a hydrogen atom and X is a methylene group: represents the compound wherein R' is a methyl group. R is a methyl group. Rja is a pentadecanovl group. is a hvdrox\ group. Ra is a hydrogen atom and X is a methylene group: exemp.comp.No.1 1 represents the compound wherein R' is a methyl group. R is a methyl group. R'S is a palmitoyl group. is a hy droxy group. is a hydrogen atom and X is a methylene group: represents the compound wherein R is a hydrogen atom. R' is a hydrogen atom. Ra is a hydrogen atom. R4, is hydroxy group. R4 is a hydrogen atom and X is a methvlene group: exemp.comp.No.49 represents the compound wherein R is a hvdrogen atom. R 2 is a methyl group. R a is a decanoyl group. is a hydroxy group. is a hydrogen atom and X is a methylene group: represents the compound wherein R is a hydrogen atom, R 2 is a methyl group. R 3 a is a lauroyl group. R 4 a is a hvdroxv group. Ra is a hydrogen atom and X is a methylene group: Doc FP9907s2.doc PS 85!FP-990o7IPCT /tsa-2ad-shsnhlish translation of spec (pages 33-85. Tables 1&2)/08.12.00 exemp.comp.No.51 represents the compound wherein R' is a hydrogen atom. RI is a methyl group. R3, is a myristoyl group. R 4 a is a hydroxy group. R'a is a hydrogen atom and X is a methylene group: exemp.comp.No.52 represents the compound wherein R' is a hydrogen atom. R 2 is methl I group. is a pentadecanol group. R a is a hxdroxv group. R' is a hydrogen atom and X is a methvlene group: exemp.comp.No.53 represents the compound wherein R' is a hydrogen atom. R 2 is a methyl group. R'a is a palmitoyl group. R 4 a is a hydroxy group. R'a is a hvdrogen atom and X is a methylene group: represents the compound wherein R is a methyl group. R is a methyl group. Ra is a hydrogen atom. R 4 a is a hydroxy group. R is a decanoyl group and X is a methylene group: exemp.comp.No.91 represents the compound wherein R' is a methyl group. R is a methyl group. Ra is a hydrogen atom. is a hydroxy group. is a lauroyl group and X is a methylene group: exemp.comp.No.92 represents the compound wherein R' is a methyl group. R- is a methyl group. R a is a hydrogen atom. is a hydroxy group. is a myristoyl group and X is a methylene group: exemp.comp.No.93 represents the compound wherein R' is a methyl group. R is a methyl group. Ra is a hydrogen atom. is a hydroxy uroup. R' is a pentadecanoyl group and X is a methylene group: exemp.comp.No.94 represents the compound \\herein R' is a methyl group. R 2 is a methvyl group. is a hydrogen atom. is a hydroxy group. RLa is a palmitoyl group and X is a methylene group: exemp.comp.No. 131 represents the compound wherein R' is a hvdrouen atom. R 2 is a meth'l group, R3, is a hydrogen atom. R 4 is a hydroxy group. R, is a decanoyl group and X is a methylene group: exemp.comp.No.132 represents the compound wherein R' is a hydrogen atom. R 2 is a methyl group. Ra is a hydrogen atom. is a hydroxy group. Ra is a lauroyl group and X is a methylene group: exemp.comp.No.133 represents the compound wherein R' is a hydrogen atom. R 2 is a methyl group, R 3 a is a hydrogen atom. is a hydroxy group. Ra is a myristoyl group and X is a methylene group: Doc: FP9907s2.doc P81485/FP-9907(PCT)/tsa-2ad-shi/En2lish translation ofspec (pages 33.85 Tables 12.00 exemp.comp.No.134 represents the compound wherein R is a hydrogen atom. R- is a methyl group. Ra is a hydrogen atom. is a hydroxy group. R'a is a pentadecanoyl group and X is a methylene group: exemp.comp.No.135 represents the compound wherein R' is a hydrogen atom. R- is a methyl group. R'a is a hydrogen atom. R 4 a is a hydroxv group. Ra is a palmitoyl group and X is a methylene group: exemp.comp.No.537 represents the compound wherein R' is a methyl group. R- is a methyl group. Ra is a hexvloxvcarbonvyl group. is a hvdroxy group. R5 is a hydrogen atom and X is a methvlene group: exemp.comp.No.538 represents the compound wherein R is a methyl group. R is a methyl group. R, is a hept iox% carbonyl group. R-a is a hvdrox% group. R is a hydrogen atom and X is a methylene group: exemp.comp.No.539 represents the compound wherein R is a methyl group. R is a methyl group. Ra', is an octyloxycarbonyl group. is a hydroxy grou p. Ri is a hydrogen atom and X is a methylene group: exemp.comp.No.540 represents the compound wherein R is a methyl group. R is a methyl group. R 3 is a nonyloxycarbonyl group. is a hydroxy group. R is a hydrogen atom and X is a methylene group: exemp.comp.No.541 represents the compound wherein R is a methyl group. R is a methyl group. Ra is a decyloxycarbony group. is a hvdroxv group. is a hydrogen atom and X is a methylene group: exemp.comp.No.542 represents the compound \wherein R is a methyl group. R is a methyl group. is an undec% loxcarhon\l igroup. R 4 is a hydroxy group. R, is a hydrogen atom and X is a meth ilene group: exemp.comp.No.543 represents the compound wherein R is a meth>i group. R- is a methyl group. R a is a dodecyloxycarbonyl group. R"a is a hydroxy group. is a hydrogen atom and X is a methvlene group: exemp.comp.No.550 represents the compound wherein R is a methyl group. R is a methyl group. R a is a hydrogen atom. R"a is a hydroxy group. R, is a hexyloxycarbonyl group and X is a methylene group: exemp.comp.No.551 represents the compound wherein R' is a methyl group. R is a methyl group, R 3 a is a hydrogen atom. R'a is a hydroxy group Ra is a heptyloxycarbonl group and X is a methylene group; Doc: FP9907s2doc P81485/FP-Q907(PCT)itsa-gad-shiEnnlish translation of spec (pages 33-85 Tables 1&2 108.12 00 exemp.comp.No.552 represents the compound wherein R is a methyl group. R is a methyl group, Ra3, is a hydrogen atom. R a is a hydroxy group, R 5 a is an octyloxvcarbonyl group and X is a methylene group: exemp.comp.No.553 represents the compound wherein R' is a methyl group. R 2 is a methyl group. R'a is a hydrogen atom. R a is a hydroxy group. R)a is a nonvloxvcarbonyl group and X is a methylene group: exemp.comp.No.554 represents the compound wherein R' is a methyl group. R is a methyl group. R3 is a hydrogen atom. R a is a hydroxv group. R is a decyloxvcarbonyl group and X is a methylene group: exemp.comp.No.555 represents the compound wherein R is a methyl group. R 2 is a methyl group. Ra is a hydrogen atom. is a hydroxy group. R, is an undecyloxycarbonyl group and X is a methylene group: exemp.comp.No.556 represents the compound wherein R' is a methyl group. R is a methyl group. is a hydrogen atom. is a hydroxy group. is a dodecyloxycarbonyl group and X is a methylene group: exemp.comp.No.563 represents the compound wherein R' is a hydrogen atom. R is a methyl group. R'a is a hexyloxycarbonyl group. R-a is a hydroxy group. R 3 is a hydrogen atom and X is a methylene group: exemp.comp.No.564 represents thecompound wherein R' is a hydrogen atom. R 2 is a methyl group. R is a heptvloxycarbonyl group. R a is a hvdroxy group. R is a hydrogen atom and X is a methvlene group: exemp.comp.No.565 represents the compound wherein R is a hydrogen atom. R 2 is a methyl group. is an octyloxycarbonyl group. is a hvdroxv group. Ra is a hydrogen atom and X is a methylene group: exemp.comp.No.566 represents the compound wherein R is a hydrogen atom. R 2 is a methyl group. Ra is a nonyloxycarbonyl group. R 4 is a hydroxy group. R' 3 is a hydrogen atom and X is a methylene group: exemp.comp.No.567 represents the compound \wherein R is a hvdrouen atom. R7 is a methyl group. R 3 is a decyloxycarbonyl group. is a hydroxy group. R, is a hydrogen atom and X is a methylene group: exemp.comp.No.568 represents the compound wherein R is a hydrogen atom. R 2 is a methyl group, R a is an undecyloxycarbonyl group, R 4 a is a hydroxy group. Wa is a hydrogen atom and X is a methylene group: Doc. FP9907s2.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 33-85 Tables 1&2)/08.12.00 exemp.comp.No.569 represents the compound wherein R' is a hydrogen atom. R' is a methyl group. R'a is a dodecyloxycarbonyl group. Ra is a hydroxy group. R a is a hydrogen atom and X is a methvlene group: exemp.comp.No.576 represents the compound wherein R' is a hydrogen atom. R is a methyl group. R a is a hydrogen atom. R is a hydroxy group. Ra is a hexvloxcarbon yl group and X is a methylene group: exemp.comp.No.577 represents the compound wherein R' is a hydrogen atom. R is a methyl group. R a is a hydrogen atom. R' is a hvdroxy group. R is a heptyloxycarbonyl group and X is a methylene group: exemp.comp.No.578 represents the compound wherein R' is a hvdrogen atom. R 2 is a methyl group. R3a is a hydrogen atom. R 4 is a h\droxy group. Ra is an octyloxycarbonyl group and X is a methvlene group: exemp.comp.No.579 represents the compound wherein R' is a hvdrouen atom. R is a methyl group. is a hydrogen atom. R is a hydroxy group. KR is a nonyloxycarbonyl group and X is a methylene group: exemp.comp.No.580 represents the compound wherein R is a hdrouen atom. R 2 is a methyl group. R3a is a hydrogen atom. is a hydroxy group. is a decylox'carbon:l group and X is a methvlene group: exemp.comp.No.581 represents the compound wherein R is a hvdrouen atom. R 2 is a methyl group. R, is a hydrogen atom. is a hidroxy group. R 3 is an undecyloxycarbonvi group and X is a methvlene group: exemp.comp.No.582 represents the compound wherein R is a hydrogen atom. R 2 is a methyl group. Ra is a hydrogen atom. R, is a hydrox\ group. is a dodecyloxycarbonyl group and X is a meth\lene group: exemp.comp.No.594 represents the compound wherein R1 is a methyl group. R is a methyl group. Ra is a decyl group. is a hydroxy group. Rh, is a hydrogen atom and X is a methylene group: exemp.comp.No.710 represents the compound wherein R is a hvdropen atom. R 2 is a methyl group. Ra is a decyl group. R is a hydroxy group. R is a hydrogen atom and X is a methylene group: exemp.comp.No.891 represents the compound wherein R is a methyl group. is a methyl group. R' is a hydrogen atom. and R 5 is a hydrogen atom: exemp.comp.No.895 represents the compound wherein R is a methyl group. R' is a mehl group. R' is a decanoyl group. and R' is a hydrogen atom: Doc: FP9907s2.doc P81485!FP-9907( PCT)/rsa-gad-sh.EnghIsh translation of spec (papes 33-.85 Tables 1&2 )08 12 00 exemp.comp.No.925 represents the compound wherein R' is a methyl group. R" is a methyl group. R 3 is a hydrogen atom. and R 5 is a decanoyl group: exemp.comp.No.1091 represents the compound wherein R' is a methyl group. R" is a dodecyl group. R' is a hydrogen atom. and R' is a hydrogen atom: exemp.comp.No. 1141 represents the compound wherein R' is a hydrogen atom. R" is a methyl group. R 3 is a hydrogen atom. and R' is a hydrogen atom: exemp.comp.No. 1145 represents the compound wherein R' is a hydrogen atom. R'' is a methyl group. R 3 is a decanoyl group. and R' is a hydrogen atom: exemp.comp.No. 1175 represents the compound wherein R' is a hydrogen atom. R'' is a methyl group. R 3 is a hydrogen atom. and R" is a decanoyl group: and exemp.comp.No.1341 represents the compound wherein R' is a hydrogen atom. R" is a dodecyl group. is a hydrogen atom. and R5 is a hydrogen atom.
Doc. FP9907s2.doc P81485/FP-9QO07(PCT )tsa-gad-shiEnglhsh translation of spec (pages 33-85 Tables 1 &2 /08.12.00 Compounds of the present invention represented by the formula or (la) can be prepared by the process as described below.
Compounds A-500359A (Exemp. compound No. A-500359C (Exemp.
compound No. A-500359D (Exemp. compound No. A-500359G (Exemp.
compound No. 45) and A-500359M-2 (Exemp. compound No. 396) of the present invention each represented by the formula are available by culturing a microorganism capable of producing the above described compounds. belonging to the Strepiomyces spp. on a suitable medium and then recovering the compound from the cultured broth. Strepiomwces griseus Strain SANK60196 (which will hereinafter be called "Strain SANK60196"). a preferable microorganism capable of producing Compounds A-500359A. A-500359C. A-500359D. A-500359G or A-500359M-2 has been collected and separated from the soil of Mt. Tsukuba./baraki-ken in a manner known to those skilled in the art.
Mycological properties of Strain SANK60196 are as follows: 1) Morphological appearance Strain SANK60196 showed morphological appearance as described below after cultivation at 28 0 C for !4 days on a medium specified by International Streptomyces Project (which \\ill hereinafter be abbreviated as "ISP") [refer to Shirling. E.B. and Gottlieb. -Int. J. Svst. Bacteriol. 16. 313-340 (1996) Observation through an optical microscope indicates that substrate mvcelia of SANK60196 are favourabli groin and branched and show vellowish grey. yellowish brown or pale olive colour, but unlike the strain belonging to .ocardi, .vpp.. does not show cleavage or zigzag extension. Aerial m\celia exhibit simple branching. The form of the spore chain is straight or curved and its chain is formed of 10 to 50 or greater spores. Observation through a scanning electron microscope shows that the spore has an oval shape and it has a smooth surface structure. The spore is 0.6-0.8 x 0.7-1.2 mm in dimension. The spore is formed only on the aerial mvcelia. Formation of sporangia. axial division of aerial mycelia. cleavage of aerial mycelia and sclerotia are not recognized.
Doc FP9907s3.doc D8 1848 FP-Q90-lPCT,'tsa-adsh/En2lIsh translation of spec (pages 86-134 i'l4 12 (Jf.
2) Growth characteristics on various culture media Growth characteristics of Strain SANK60196 on an agar medium after cultivation at 28 0 C for 14 days is as described below in Table 3. In the Table. the composition of the medium attached with ISP No. is the same as specified by ISP. In the item. abbreviations G. AM. R and SP stand for growth, aerial mycelia. reverse colour and soluble pigment. respectively. The colour tone is described in accordance with "*Colour Standards. ed. by Japan Colour Laboratory". The indication of the colour tone in parentheses is a colour number in accordance with Munsell colour system. The pale yellow soluble pigment produced in a water-agar medium changes into colourless by 0.05N hydrochloric acid. but shows no change by 0.05N sodium hydroxide.
[Table 3] Nature of Medium: Item: characteristics Yeast extract malt extract agar (ISP 2): G: Excellent. flat. vellowish brown (10YR 5/6) AM: Abundantly formed. velvety, pale brown (2.5Y 8'2) R: Yellowish brown 10YR 5'8) SP: Yellowish brown OYR 6'8) Oat meal agar (ISP 3): G: Excellent. flat. vellowish brown 12.5Y 6:6) AM: Abundantly formed. velvet\. pale yellowish orange (5Y 9 2) R: Dark yellow (2.5Y 8 8) SP: Not produced Starch inorganic salt agar (ISP 4: G: Good. flat. vellowish brown (2.5Y 6 4) AM: Abundantly formed, velvety. yellowish grev (7.5Y 9 2) R: Yellowish brown (2.5Y 6 4) Glycerin asparagine agar (ISP G: Excellent. flat. pale yellowish brown (2.5Y 7/6) AM: Abundantly formed, velvety. yellowish grey (5Y 8/2) R: Pale yellowish brown (2.5Y 8/6) SP: Not produced Peptone yeast extract iron agar (ISP 6); Doc FP9907s3.doc P81485/FP-9907(PCT)/rsa-ead-sh/Eneglsh translation of spec (pages 86-134 1,14 12 00 G: Excellent. flat. pale olive color (5Y 8.3) AM: Slightly produced. velvety. yellowish grev (5Y 9/1) R: Pale yellow (5Y 8/6) SP: Not produced Tyrosi ne agar (ISP 7) G: Good. flat. gravish yellow brown (15SY 5/4) AM: Abundantly formed. -velvety, light olive greV (7.5Y 8'2) R: Yel lowish brow-n (lIO'xR 5 /4) SP: Grayish \'ellow brown (2,5Y 4!3) Sucrose nitrate agar: G: Not so good. flat. pale .ellowk (5Y 8 6) AM: Abundantly formed. velvety. ILht olive Lrev (7.5Y1 82-) R: Dark yellow (5Y '8) SP: Pale yellow 9 6) Glucose asparagine agar: G: Good. flat. pale yellow 93 AM: Not so good. velvety. Nellowish Lrev (5Y 9./I) R: Yellow ish 2rev (7 5 9) SP:. Not produced Nutrient agar (product of Difco Laboratories) G: Good. flat. pale yellowish brown 2.Y8'S') AM: Good. velvety. %ellow.ish Lrev t R: Yellowish 2.re% 0 Q SP: Not produced Potato extract carrot extract aaar: G: Not so goiod. flat. %ellowish 2rev V.Y92) AM: Not so g-ood. el% etv. %ellow\ish gre% )(5Y 92 R: Yellowish arey 5Y 93 SP: Yellowish 2rey Q 5 Y 93 WVater aear: G: Not good. flat. yellowish grey (5Y 9.11 AM: Not good, vel-vety. yellowish grey (5ZY 9/1) R: Yellowish grey (7.5Y 9/4) SP: Pale yellow (5Y 9/6) Doc, FP9907s3.doc Doc FP9Os3.oc P8l485rFP-99O'(PCT)tsa-2ad-sh/English translation ofspec (pages 86-134)'14 1200 3) Physiological characteristics The physiological characteristics of the present strain observed for 2 to 21 days after cultivation at 28°C are as shown in Table 4. In the table. Medium I is a yeast extract malt extract agar medium (ISP 2).
[Table 4] Hydrolysis of starch Liquefaction of gelatin Reduction of nitrates Coagulation of milk Peptonization of milk Formation of melamine-like pigment Substrate decomposition: casein t rosine xanthine Growth temperature range (ledium 1 Optimum growth temperature (Medium 1) Growth in the presence of salt (Medium 1) positive positive positive negative positive positive positive positive negative 6 to 18 to 10 O Utilisation of a carbon source b\ Strain SANK601%6 observed after cultivation at 28 0 C for 14 days on a Pridham-Gottlieb agar medium (ISP 9) is as described in Table 5. In the table. means utilisable. while means nonutilisable.
[Table D-glucose L-arabinose D-xvlose Inositol D-mannitol D-fructose L-rhamnose Sucrose Raffinose Control Doc FP9 07s3.doc P8l485/FP-9907 PCT)i/sa.Eadsh/En2h sh translation ofspec (pages 86-3i-U414 12 00 4) Chemotaxonomic properties The cell wall of the present strain was investigated in accordance with the method of Hasegawa. et al. [refer to Hasegawa. et al.. "The Journal of General and Applied Microbiology. 29. 319-322(1983)]. resulting in the detection of LLdiaminopimelic acid. The main sugar component in the whole cells of the present strain was investigated in accordance with the method of M.P. Lechevalier [refer to Lechevalier. "Joural of Laboratory and Clinical Medicine. 71. 934-944(1 968)].
As a result. no characteristic component was detected.
The above-described mvcological properties have revealed that the present strain belongs to Streptomn.ces spp. among the actinomycetes. It has been made clear that the present strain is markedly related to Strepiomyces griseus. as a result of comparison with the microorganism described in the ISP strains by Shirling and Gottlieb [refer to Shirling. E.B. and Gottlieb. "International Journal of Systematic Bacteriology. 18. 68-189 and 279-392 (1968): 19. 391-512 (1969): 22. 265-394 the microorganism described in "The actinomvcetes Vol. 2" written by Waksman [refer to Waksman. "The actinomycetes 2 with the microorganism described in Bergey's Manual edited by Buchanan and Gibbons [refer to R.E. Buchanan and N.E. Gibbons. "Bere\ 's Manual of Determinative Bacteriology". 8th edition I 1974 ith the microorganism described in "Bergey's Manual of Systematic Bacteriology". edited b\ Williams [refer to Williams. et al.. "Bergey's Manual of S stematic Bacteriolog\ 4 (1989)"] and with the microorganism described in the recent literature about actinomycetes belonging to Sirepomvces spp. It has ho\ e' er been recognized to be different from Strecpwmyces griseus. because it produces a cllo ish grey soluble pigment on a glycerin asparagine agar medium and a pale \ello\\ish brown soluble pigment on a peptone yeast extract iron agar medium but produces a soluble pigment neither on a potato extract carrot extract agar medium nor on a w\ater agar medium: the maximum growth temperature is 40'C: and it is grown in the presence of 70o of salt.
The present strain having such mycological characteristics is considered to be a novel strain different from Streptomyces griseus. but it is impossible to distinguish them based on only the above-described differences. The present inventors therefore identified the present strain as Streptomvces griseus SANK60196.
Doc: FP9907s3.doc P81485/FP-990"PCT)!tsa-gad-sh/,Enelsh translation of spec (pages 86-134 '14 12 00 This strain was internationally deposited with Agency of Industrial Science and Technology. Ministry of International Trade and Industry Higashi 1-chome.
Tsukuba-shi. Ibaraki-ken. 305. JAPAN) as of February 22. 1996. with the accession number of FERM BP-5420.
A description was heretofore made on Strain SANK60196. It is known that various properties ofactinomycetes are not fixed but easily change naturally or synthetically. The strain usable in the present invention embraces all of such variants.
In other words. the present in\ention embraces all the strains belonging to the Strepromyces spp. and capable of producing Compounds A-500359A. A-500359C. A- 500359D. A-500359G or A-5003591-2.
Any synthetic or natural medium can be used for cultivation for microorganisms capable of producing Compounds A-500359A. A-500359C. A- 500359D. A-500359G or A-500359.1-2 of the present invention, insofar as it contains, as needed. a substance selected from carbon sources. nitrogen sources.
inorganic ions and organic nutrition sources.
Known carbon sources. nitrogen sources and inorganic salts conventionally employed for cultivation of the strain of the eumycetes or actinomycetes and are utilisable by a microorganism can be used as such nutrition sources.
Specific examples of the carbon source include glucose. fructose. maltose.
sucrose. mannitol. glycerol. dextrin. oats. rye. corn starch. potato. corn meal. soybean meal. cotton seed oil. thick malt syrup. theriac. soybean oil. citric acid and ;artaric acid. They may be used either singly or in combination. The amount of the carbon source to be added usually varies. but not limited to. within a range of from 1 to wt.%.
As the nitrogen source. a substance containing protein or hydrolyzate thereof can usually be employed. Preferred examples of the nitrogen source include soybean meal. wheat bran. peanut meal. cotton seed meal. casein hydrolyzate. Farmamine. fish meal. corn steep liquor. peptone. meat extract. pressed yeast. dry yeast. yeast extract.
malt extract, potato. ammonium sulfate. ammonium nitrate and sodium nitrate. It is preferred to use the nitrogen source either singly or in combination in an amount ranging from 0.2 to 6 wt.% of the amount of the medium.
Doc FPQ907s3.doc P81485/FP-9907'PCT i,'sa-gad-sh!Enghsh translationofspec (pages 86-1341.14 12 00 As the nutrition inorganic salt. ordinarily employed salts from which an ion is available, such as sodium salts. ammonium salts. calcium salts, phosphates. sulfates.
chlorides and carbonates can be used. In addition, trace metals such as potassium.
calcium. cobalt. manganese. iron and magnesium are usable.
For the production of Compound A-500359A. the addition of cobalt or yeast extract is particularly effective.
Upon culturing the microorganism capable of producing Compound A- 500359A. A-500359C. A-500359D. A-500359G or A-500359M-2. an inhibitor of antibiotic biosynthesis can be added to produce useful related compounds.
Compound A500359M-2 can be produced. for example. by using. as a medium additive. S-(2-aminoethyl -L-cvsteine or salt thereof which is an aspartate kinase inhibitor. The additive can be added to give its final concentration ranging from 1 to 100 mM. Preferably. use of it to give a final concentration of 10 mMl permits favorable production of Compound A-5003591\-2.
Upon liquid culture. a silicone oil. vegetable oil or surfactant can be added as an antifoamer.
The medium used for the cultivation of Strain SANK 60196 to produce Compound A-500359A. A-500359C. A-500359D. A-500359G or A-500359M-2 preferably has a pH ranging from 5.0 to The temperature which allows Strain S.NK60196 to grow ranges from 12 to 36°C. It is preferred to culti\ate the strain at IS to 28:C in order to produce Compound A-500359A. A-50O3 S C. A-500359D. A-500359G or A-500359M-2. of which 19 to 23C is more preferred.
Compound A-500359A. A-500359C. A-500359D. A-500359G or A- 500359M-2 is available by aerobic culture of Strain SANK 60196. Ordinaril\employed solid culture. shake culture. and aeration agitation culture can be used as such culturing method.
For small-scale culturing. agitation of the culture for several da\s at 1 to 23°C is preferred. Culturing is started by growing a seed culture in a single or two stage process in an Erlenmeyer flask equipped with a baffle (water flow adjusting wall) or an ordinarily-employed Erlenmever flask. A carbon source and a nitrogen source can be used in combination as a medium in the seed culture. The flask or seed culture may be shaken at 19 to 23°C for 5 days or until the seed cultures grow Doc- FP9907s3.doc P81 485/FP-9907 PCT I.rsa-2ad-sh/English translation of spec (pages 86-134 i,14 12 00 sufficiently in a thermostat incubator. The seed cultures thus grown can be used for inoculation of the second seed culture medium or a production medium. When the seed cultures are used under an intermediate growing step. they are allowed to grow essentially in a similar manner. followed by inoculation of a part of them into a production medium. The flask into which the seed cultures has been inoculated is subjected to culturing with shaking at a constant temperature for several days and after completion of the culturing. the cultured medium in the flask is centrifuged or filtered.
For large-scale cultivation, on the other hand. culturing in a jar fermenter or tank equipped with an agitator and an aeration apparatus is preferred. Prior to culturing in such a container, the culture medium is heated to 125CC for sterilization.
After cooling, the seed cultures which have been allowed to grow in advance by the above-described method are inoculated on the sterilized medium. Then. culturine is carried out with aeration and acitation at 19 to 23"C. This method is suitable for obtaining a large amount of compounds.
Compound A-500359NI-2 can be produced by adding, as an aspartate kinase inhibitor, an aqueous solution of S-(2-aminoethyl)-L-cysteine or salt thereof which has been filter sterilized in advance to a sterilized medium at the beginning time of the cultivation or durine cultivation.
The production of Compound A-500359.A. A-500359C. A-500359D. A- 500359G or A-500359M\-2 produced can be measured by sampling a portion of the cultured broth and subjecting it to high performance liquid chromatography. The titre of Compound A-500359.A. A-500359C. A-500359D. A-500359G or A-500359M-2 usually reaches a peak in 3 to 9 days.
After completion of the cultivation, the cell component is separated from the cultured broth by separation with the aid of diatomaceous earth or centrifugation.
Compound A-500359A. A-500359C. A-500359D. A-500359G or A-500359M-2 present in the filtrate or supernatant is purified by utilizing its physico-chemical properties with HPLC analytical data as an index. Compound A-500359A. A- 500359C. A-500359D. A-500359G or A-500359M-2 present in the filtrate can be purified by using adsorbents singly or in combination, such as activated charcoal (product of Wako Pure Chemicals) and an adsorbing resin such as "Amberlite XAD-2 or XAD-4" (trade name; product of Rohm Haas). and "Diaion HP-10. HP-20. CHPor HP-50. Sepabeads SP205. SP 2 06 or SP207" (trade name: product of Doc. FP9907s3.doc P81485/FP-9907 1 PCT)/tsa-gad-sh/English translation of spec (pages 86-134 /14.12 00 Mitsubishi Chemical). Compound A-500359A. A-500359C. A-500359D. A- 500359G or A-500359M-2 in the solution can be separated from impurities by passing a solution containing them through the layer of such adsorbents. or by eluting the adsorbed compounds from the layer \with aqueous methanol. aqueous acetone or aqueous normal butanol.
Compounds A-500359A. A-500359C. A-500359D. A-500359G or A- 500359M-2 thus obtained can be purified by adsorption column chromatography using an adsorbent such as silica gel. "Florisil" (trade name), or "Cosmosil" (trade name: product ofNacalai Tesque): partition column chromatography using "Sephadex (trade name: product of Pharmacia Biotech): gel filtration chromatography using "Toyopearl HW40F (trade name: product of TOSOH Corp): or high performance liquid chromatographv using a normal phase or reversed phase column: or the like.
Compounds A0059. -500359. A-500359C. A-500359D. A-500359G or A- 500359M-2 according to the present invention can be separated and purified by using the above-exemplified separation and purification means singly or in combination as needed. or in some cases. by using one of them in repetition.
Compounds A-500359A. A-500359C. A-500359D. A-500359G and A- 500359M-2 of the present invention thus obtained are novel compounds not published in the literature but their antibacterial activit\ can be determined by a method known to those skilled in the art.
Ester derivatives. ether derivatives and N-aikvlcarbamovl derivatives can each be prepared easily by using an\ one of the below-described Processes A to F or using them in combination as necessar\.
(Process A) Process A is for the preparation of an ester derivative of Compound (la) and by this process. Compound (lc) wherein R" is a methyl group can be prepared.
Doc FP9907s3.doc P81485/FP-9907(PCTi'tsa-ead-sEnglish translation of spec (pages 86-134' 14 12 00 Process A Step Al1
HN-
R
OH
(11) 0
H
HI/N
N
R- -I OR 5 1
-OH
yO O 0
NH
2 r 0 Step A2 CH-~O
OR
3 t (111)
OF
"OR 4
CON
0 0 0O -0
R
xCHIO
OR
3 (1c) wherein: Rl and X have the same meaninus as described above. represents a h-ydrogen atom or a hx-drox protecting group. WRepesnt hvdrouen atom. a hvdroxv- protecting group or an ester residue. R 4 b represents a hydrogen atom. a hydroxv-protecting group or an ester residue. RWb represents a hydrogen atom or a hydroxv- protecting group. and R 5 c represents a hydrogen atom. a hydroxv -protecting group or an ester residue. with the proviso that R 3 b and R'b do not represent a Doc FP9907s3.doc Doc F99O~s.doc P81 485/FP-QQ07i PCT)ttsa-gad-sh/En2Iish translation of spec (pages 86-134)/14 1200 hydrogen atom at the same time and Rb and do not all represent a hydrogen atom or a h-vdroxv -protecting group at the same time.
Step AlI is for the preparation of a compound having the formula (III) and it is accomplished by protecting the hydroxy group of the compound of formula (I1).
Although the hydroxy-protecting step differs depending on the kind of the protecting group. it is conducted by a process well k-nown in synthetic organic chemistry.
When the hydroxy -protecting group is a -sllgroup"'. "alkoxymethyl group".
substituted ethyl group". "aralkvl group-. -alkoxvcarbony I group".
,-al ken vloxyvcarbonyl group**. "aralkyloxy carbonyl group". l-(aliphatic ac\lox\lower alkvl group". -aliphatic acylthio)-lower alkyl group-. ~l (c\vcloalk\lcarbonvl-ox\v)-lo\\ er alkyl group". -1 -(aromatic acvloNVi-low er alk\ I group-. I -(lower alkoxycarbon% lox% )-lower alkyl group-. -1cycloalkyloxycarbonylox\ -lower alkyl group". "phthalid\ I group".
-"oxodioxolenvlmrethyl uroup-. -"carbamoyi group substituted wilth 2 lower alkVl uroups"*. "I -(lower alkoxvcarbonvlox% )-lower alkyl group". "lower alkyl-dlithio ethyl group" or I -(acvloxv )-alkvlox% carbon vI group-. this step is conducted by reacting Compound (11) with a desired h% drox protecting group halide in an inert solvent in the presence of a base.
Examples of the hvdrox% -protecting group halide usable in the above reaction include trimethylsilyl chloride. irieth\ lsiR% I chloride. t-but\ ldimethylsilvl chloride. tbutvilimethvili\Il bromide. meth\ ldi-t-butv lsl\ I chloride. methvil-t-hutvl1silvl bromide. diphenylmethyisil\ c1rie diphenvlm-eth\ IslivI bromide. methoxym eth% I chloride. 2-methoxvethox\ rneth% I chloride. 2-.2.2'-trichloroethoxymeithl\I chloride. I ethoxvethvl chloride. benz% 1 chloride. benz% I bromide. ci-naphth\ lmcth% I chloride.
diphenylmethyl chloride. diphe:n\ ime th\ I bromide. triphenylreth\ I chloride. 4methylbenzyl chloride. 4-methON% benz\ I chlorideC. 4-nitrobenz\ I chloride. 4 chlorobenzyNl chloride. methoN~carbonyl chloride. ethoxvcarbonyl chloride. 222 trichloroethoxycarbonyl chloride, vinyl oxycarbonyl chloride. allylox~carbonyl chloride. benzvloxNvcarbon,,l chloride. benzylovc arbonyl bromide. 4methoxvbenzy Iox tcarbon\vl chloride. 4 -nitrobenzyloxycarbonyl chloride.
acetoxv~methyl chloride. propi ony loxvymethyl chloride. butvrylox%!methyl chloride.
pivaloyloxymethyl chloride. pivaloyloxymethyl bromide. valenwloxym ethyl chloride.
Doc. FP9907s3.doc Do. P907~dc P81 485/FP-9907 POT /tsa-2ad-shlEne I sl, IranslIat ion of spec (pages 86-134 )14 1200 I -acetoxvethyl chloride. but, ryloxyethvl chloride. I -pivaloyloxyethyl chloride, cv~c lopentylcarbonyloxymethyl chloride. cyc lohexylIcarbonylIoxymethylI chloride. Icy~clopentyl1carbonylIoxyethyl. chloride. I -cv clohexylIcarbon\vloxveth\vl chloride.
methoxv-carbonvloxvmethvl chloride. methox vcarbon vloxvmeth vl bromide.
ethoxycarbonyloxymethyl chloride. propoxycarbonyloxymethyl chloride.
isopropoxycarbonvloxymeth \l chloride. butoxycarbonyloxymethyl chloride.
isobutoxycarbonyloxymethyl chloride. Il-(methoxv-carbon\-lox\-)ethv-l chloride. I (methoxNvcarbolvloxY)ethy-l bromide. I ethox vcarbon yloxy)ethvl chloride. I (isopropoxycarbonylIoxy')ethylI chloride. cc lopenty loxy*carbony lox ymethylI chloride.
cvclohexvlIox\vcarbonylIoxy-methylI chloride. I cvclopentvlIoxvcarbon\vlx\ )ethvl) chloride. I -(cc lohexy l ox' ycarbon yloxv %ethvl] chloride. phthalidyl chloride. phthalidlyl bromide. (5,-phenv->-'oxo- I .3-dlioxolen-4-yhl methyl chloride. [5-(4-methylphenyl P2oxo- I .3-dioxolen-4-yIlmeth\ I chloride. 5--methv-2-oxo-I .3-dioxolen-4-yl Imneth\vl chloride. (5-methvl-2-oxo- I .S'-dioxolen-4-vl hmethyl bromide. (5-eth\ dioxolen-4-vl)meth\vl chloride. dimethvlcarbamoN-l chloride. diethvlcarbamnovl chloride. methyvlithioethyl chloride. ethvldithioethNvl chloride and piv~alo\vloxy'methvloxy,,carbon\vl chloride, of'which triethylsilyl chloride. tbutvldimethvlsilvl chloride. t-butvldimethvlsilv bromide. benzvl chloride. benzNvl bromide. triphenlmethyl chloride. 4-methox\'benzyl chloride. 222 tri c hloroethox vcarbon chloride. al lyloxy carbonyl chloride. benzylIox% carbonyl chloride. benz%-lox\vcarbonylI bromide. acetoxvm ethyl chloride and pivalo\ loxymethyl chloride are preferred.
Examples of the base include alkali metal hydroxides such as lithium h%-droxide, sodium hydroxide and potassium hydroxide. alkali metal carbonates such as lithium carbonate, sodium carbonate and potassium carbonate. alkali metal bicarbonates such as sodium bicarbonate and potassium bicarbonate. alkali metal alkoxides such as lithium methoxide. sodium methoxide. sodium ethoxide and potassium t-butoxide. and organic amines such as triethylamine. tribut% lamine. Nmethylmorpho line. pyridine. 4-dimeth\laminopyridine. picoline. lutidine. collidine.
I .5-diazabicyclo[4.3'.O]-5 -nonene and I .8-diazabicy-clo[5.4.O].7-undecene. Out of these. org-anic amines are preferred. of which triethylamine. tributvlamine. pyridine and lutidine are particularly preferred. Upon use of an organic amine in the liquid form. it also serves as a solvent when used in large excess.
Doc FP9<)07s3.doc Doc F9~O~~doc P81 485/FP-9907 (PCT1!rtsa-ead-shlEneiish translation of spec (paecs 86-134),14 12 00 98 There is no particular limitation on the inert solvent used in the above reaction, provided it is inert to the reaction. Examples include hydrocarbons such as hexane, benzene and toluene, halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride and 1,2-dichloroethane, ethers such as ether, tetrahydrofuran and dioxane, ketones such as acetone and methyl ethyl ketone, nitriles such as acetonitrile, amides such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethylsulfoxide; and mixtures thereof. Of these, hydrocarbons and amides are preferred.
Although the reaction temperature differs with the nature of the starting compound the halide and the solvent, it usually ranges from -10 0 C to 100°C (preferably 0 to 50 0 Although the reaction time differs with the reaction temperature or the like, it ranges from 30 minutes to 5 days (preferably i to 3 days).
When the hydroxy-protecting group is a "tetrahydropyranyl or tetrahydrothiopyranyl group" or a "tetrahydrofuranyl or tetrahydrothiofuranyl group", Compound (II) is reacted with a cyclic ether compound such as dihydropyran, 3bromodihydropyran, 4-methoxydihydropyran, dihydrothiopyran, 4methoxydihydrothiopyran, dihydrofuran or dihydrothiofuran in an inert solvent in the presence of an acid.
Examples of the acid usable in the above reaction include inorganic acids such as hydrogen chloride, nitric acid, hydrochloric acid and sulfuric acid and organic acids such o as acetic acid, trifluoroacetic acid, methanesulfonic acid and p-toluenesulfonic acid, of which hydrogen chloride, hydrochloric acid, sulfuric acid and trifluoroacetic acid are preferred, with hydrogen chloride and hydrochloric acid being particularly preferred.
S. Examples of the inert solvent usable in the above reaction (which is inert to the reaction) include hydrocarbons such as hexane, benzene and toluene, halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride and 1,2dichloroethane, ethers such as ether, tetrahydrofuran and dioxane, ketones such as acetone and methyl ethyl ketone, nitriles such as acetonitrile, amides such as N,Ndimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethylsulfoxide; and mixtures thereof. Of these, hydrocarbons and ethers are preferred.
Doc: FP9907al.doc P814851FP-9907(PCTtsa/gad/shjcofected pagcs of spccIO3IOI01 Although the reaction temperature differs with the nature of the starting compound the cyclic ether compound and the solvent, it usually ranges from 0 C to 100 0 C (preferably 0 to 50C). Although the reaction time differs with the reaction temperature or the like. it usually ranges from 30 minutes to 5 days (preferably 1 to 3 days).
When the hydroxy-protecting group is a "carbamoyl group" or "carbamovl group substituted with one lower alkyl group". Compound (II) is reacted with an isocvanate or lower alkyl isocyanate such as methyl isocyanate or ethyl isocyanate in an inert solvent in the presence or absence of a base.
Preferred examples of the base usable in the above reaction include the aboveexemplified organic amines. with triethylamine. tributylamine. pyridine and lutidine being particularly preferred.
There is no particular limitation on the inert solvent used in the above reaction provided that it is inert to the reaction. Examples include hydrocarbons such as hexane. benzene and toluene. halogenated hydrocarbons such as dichloromethane.
chloroform, carbon tetrachloride and 1.2-dichloroethane. ethers such as ether.
tetrahydrofuran and dioxane. ketones such as acetone and methyl ethyl ketone. nitriles such as acetonitrile. amides such as N.N-dimethylformamide. N.Ndimethylacetamide. N-methyl-2-pyrrolidone and hexamethylphosphoramide. and sulfoxides such as dimethvlsulfoxide: and mixtures thereof. Of these. hydrocarbons and ethers are preferred.
Although the reaction temperature differs with the nature of the starting compound the cyclic ether compound and the solvent, it usuall\ ranges from to 100'C (preferably 0 to 5S0C). Although the reaction time differs with the reaction temperature or the like. it ranges from 30 minutes to 5 days (preferably 1 to 3 days).
After completion of the reaction, the desired compound in each reaction is collected from the reaction mixture in a manner known to those skilled in the art. The desired compound can be obtained. for example. by filtering off any insoluble matter.
as required, and then distilling off the solvent under reduced pressure: or by distilling off the solvent under reduced pressure, adding water to the residue. extracting the mixture with a water immiscible organic solvent such as ethyl acetate. drying over anhydrous magnesium sulfate or the like and then distilling off the solvent. If necessary, the resulting product can be purified further in a manner known to those Doc. FP9907s3.doc P81485/FP-Q907(PCT)/tsa-gad-shiEnglish translation of spec (pages 86-134 /14 12 00 skilled in the an. for example. by recrystallization. column chromatography or the like.
Step A2 is for the preparation of a compound having the formula This step can be accomplished by esterifying Compound (III) and if desired. removing the hydroxy-protecting group from the esterified compound.
Esterification is conducted by reacting Compound (III) with an acid halide or acid anhydride having a desired ester residue in an inert solvent in the presence of a base.
Examples of the acid halide or acid anhydride used in the above reaction include compounds represented by any one of the formulae R 6 CO-Y. RoCOCOR.
R CO-O-COR 6 and R"OCO-Y [wherein R" has the same meaning as described above.
Y represents a halogen atom. preferably chlorine or bromine. R 9 represents a C-, alkyl group (preferably ethyl or isopropyl)]: a mixed acid anhydride of formic acid and acetic acid. cyclic acid anhydrides such as succinic acid anhydride. glutaric acid anhydride and adipic acid anhydride: and phosphate ester introducing agents such as compounds represented by the formula (R O):PO-Y (wherein Y has the same meaning as described above and R represents a lower alkyl group). of which the compounds represented by an\ one of the formulas ROCO-Y. R 6
COCOR
0
R
6
CO-O-
COR
6 and R OCO-Y (wherein Y and RQ ha\ e the same meaninus as described above) are preferred.
Examples of the base usable in the abo\ e reaction include alkali metal hydroxides such as lithium hydroxide, sodium h\droxide and potassium hydroxide.
alkali metal carbonates such as lithium carbonate. sodium carbonate and potassium carbonate. alkali metal bicarbonates such as sodium bicarbonate and potassium bicarbonate. alkali metal alkoxides such as lithium methoxide, sodium methoxide.
sodium ethoxide and potassium t-butoxide. and oruanic amines such as triethvlamine.
tributylamine. N-methylmorpholine. pyridine. 4-dimethylaminopyridine. picoline.
lutidine. collidine. 1.5-diazabic\ clo[4.3.0]-5-nonene and 1.8-diazabicyclo[5.4.0]-7undecene. Of these, organic amines are preferred. of which triethvlamine.
tributylamine. pyridine and lutidine are particularly preferred. Upon use of an organic amine in the liquid form. it also serves as a solvent when used in large excess.
When the esterifying reaction is a phosphate ester introducing reaction. it can also be conducted by reacting Compound (III) with a phosphite having a desired ester Doc FP9907s3.doc P81 485/FP-9Q0Q PCT),sa-gadsh/En2lIsh translation of spec (pages 86-1 34)'14 12 00 101 residue in an inert solvent in the presence of an acid or base, and oxidizing the reaction mixture into the corresponding phosphate ester by an oxidizing agent.
As the phosphite, a compound represented by the formula (R 7 0) 2 wherein R 7 represents a C6-20 alkyl group and Z represents a halogen atom or a compound represented by the formula -N(Rg)2 (wherein R 8 represents a lower C- 20 alkyl group)] can be used.
When, in the above formula, Z represents a halogen atom, a base is employed as a catalyst and examples of the base usable are similar to those exemplified above. When Z is not a halogen atom, on the other hand, an acid is used as a catalyst. Any acid can be used, provided that it exhibits acidity as strong as acetic acid. Tetrazole is preferred.
Examples of the oxidizing agent usable in the above reaction include metachloroperbenzoic acid, t-butylhydroperoxide and peracetic acid, of which metachloroperbenzoic acid is preferred.
There is no particular limitation on the inert solvent usable in the above reaction, provided that it is inert to the reaction. Examples include hydrocarbons such as hexane, benzene and toluene, halogenated hydrocarbons such as dichloromethane, chloroform, carbon tetrachloride and 1,2-dichloroethane, ethers such as ether, tetrahydrofuran and dioxane, ketones such as acetone and methyl ethyl ketone, nitriles such as acetonitrile, *amides such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2- S0. pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethylsulfoxide; and mixtures thereof. Of these, hydrocarbons and amides are preferred.
Although the reaction temperature differs with the nature of the starting compound (III), the phosphite and the solvent, it usually ranges from -10°C to 100 0
C
(preferably 0 to 50 0 The reaction time differs with the reaction temperature and the like, but it ranges from 10 minutes to 2 days (preferably 30 minutes to 10 hours).
Esterification can also be conducted by reacting Compound (111) with a carboxylic acid having a desired ester residue in an inert solvent in the presence of a condensing agent.
Examples of the condensing agent usable in the above reaction include So: carbodiimides such as dicyclohexylcarbodiimide, carbonyl diimidazole and 1-(N,Ndimethylaminopropyl)-3-methylcarbodiimide hydrochloride, of which dicyclohexylcarbodiimide is preferred.
Doc: FP9907al.doc P81485/FP-9907(PCT) sa/gad/sh/corrected pages of sped03/01/01 There is no particular limitation on the inert solvent used in the above reaction.
provided that it is inert to the reaction. Examples include hydrocarbons such as hexane. benzene and toluene. halogenated hydrocarbons such as dichloromethane.
chloroform, carbon tetrachloride and 1.2-dichloroethane. ethers such as ether.
tetrahydrofuran and dioxane. ketones such as acetone and methyl ethyl ketone. nitriles such as acetonitrile. amides such as N.N-dimethylformamide. N.Ndimethylacetamide. N-methyl-2-pyrrolidone and hexamethylphosphoramide. and sulfoxides such as dimethylsulfoxide: and mixtures thereof. Of these. hydrocarbons.
halogenated hydrocarbons and amides are preferred.
Although the reaction temperature differs with the nature of the starting compound (III). carboxylic acid and solvent, it usually ranges from -10-C to 100'C (preferably 0 to 50'C). The reaction time differs with the reaction temperature or the like. but it usually ranges from 10 minutes to 2 days (preferably 30 minutes to hours).
After completion of the reaction, the desired compound in each reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by filtering off any insoluble matter, as necessary, and then distilling off the solvent under reduced pressure: or by distilling off the solvent under reduced pressure, adding water to the residue.
extracting the mixture with a water immiscible organic solvent such as ethyl acetate.
drving over anhydrous magnesium sulfate or the like and then distilline off the solvent. If necessary, the resulting product can be purified further in a manner known to those skilled in the art, for example. by recrystallization. column chromatography or the like.
Although the desired deprotection of hydroxy-protecting group differs with the kind of protecting group. it is conducted by the process well known in synthetic organic chemistry.
When the hydroxy-protecting group is an "aralkvl group" or "aralkyloxycarbonyl group". deprotection is conducted by contactine the corresponding compound with a reducing agent (including catalytic reduction) or oxidizing agent in an inert solvent.
There is no particular limitation on the inert solvent usable in the removal by catalytic reduction, provided that it is inert to the reaction. Examples include alcohols such as methanol and ethanol. ethers such as diethyl ether, tetrahydrofuran and Doc FP9907s3.doc P81485/FP-9907 PCT),;tsa-gad-shEnlish translafion of spec (pages 86-13414 200 dioxane. aromatic hydrocarbons such as toluene. benzene and xvlene and aliphatic hydrocarbons such as hexane and cyclohexane and esters such as ethyl acetate and propyl acetate and aliphatic acids such as acetic acid: and mixtures of the aboveexemplified organic solvent and water. of which alcohols are preferred.
Although there is no particular limitation on the catalyst usable in the above reaction (provided that it is ordinarily employed for catalytic reduction). examples include palladium on carbon. Raney nickel. platinum oxide, platinum black. rhodiumaluminium oxide. triphenylphosphine-rhodium chloride and palladium-barium sulfate.
of which palladium on carbon is preferred.
Although there is no particular limitation on the pressure of hydrogen, it usually ranges from 1 to 10 times atmospheric pressure (preferably 1 to 3 times atmospheric pressure).
Although the reaction temperature or reaction time differs with the nature of the starting substance. the solvent and the catalyst, the reaction temperature usually ranges from -20^C to 100C (preferably 0 to 50'C) and the reaction time usually ranges from 30 minutes to 10 hours (preferably 1 to 5 hours).
There is no particular limitation on the inert solvent usable upon deprotection by an oxidizing agent. provided that it is inert to the reaction. Examples include ketones such as acetone. halogenated hydrocarbons such as methylene chloride.
chloroform and carbon tetrachloride. nitriles such as acetonitrile. ethers such as diethyl ether. tetrahvdrofuran and dioxane. amides such as dimethylformamide.
dimethylacetamide and hexamethylhposphoramide and sulfoxides such as dimethylsulfoxide. and mixed solvents thereof. Preferred are the amides and sulfoxides.
There is no particular limitation imposed on the oxidizing agent usable in the above reaction, provided that it ma\ be employed for oxidization. Examples include alkali metal persulfates such as potassium persulfate and sodium persulfate. ceric ammonium nitrate (CAN) and 2.3-dichloro-5.6-dicyano-p-benzoquinone (DDQ). of which ceric ammonium nitrate (CAN) and 2.3-dichloro-5.6-dicyano-p-benzoquinone (DDQ) are preferred.
Although the reaction temperature and reaction time differs with the nature of the starting substance. the solvent and the catalyst, the reaction temperature usually ranges from -10 0 C to 150 0 C (preferably 0 to 50 0 C) and the reaction time usually ranges from 10 minutes to 24 hours (preferably 30 minutes to 10 hours).
Doc FP9907s3.doc P81 485/FP-9907( PCTVtsa-gad-sh/English iranslation of spec (pages 86-134 '14 12 00 When the hydroxy-protecting group is a t-butyl roup. t-butoxycarbonyl group. "alkoxymethyl group". "tetrahvdrop.ranyl or tetrahydrothiopvranyl group" or "tetrahvdrofuranyl or tetrahydrothiofuranyl group". deprotection is conducted by reacting the corresponding compound with an acid in an inert solvent.
There is no particular limitation on the inert solvent used in the above reaction.
provided that it is inert to the reaction. Examples include hydrocarbons such as hexane and benzene. halogenated hydrocarbons such as methylene chloride.
chloroform and carbon tetrachloride. esters such as ethyl acetate. ketones such as acetone and methyl ethyl ketone. alcohols such as methanol and ethanol. ethers such as ether. tetrahydrofuran and dioxane: and mixtures thereof with water. Of these.
esters. ethers and halogenated hydrocarbons are preferred.
Examples of the acid usable here include inorganic acids such as hydrogen chloride, nitric acid. hydrochloric acid and sulfuric acid. organic acids such as acetic acid. trifluoroacetic acid. methanesulfonic acid and p-toluenesulfonic acid and Lewis acids such as boron trifluoride. of which the inorganic acids and organic acids are preferred and hydrochloric acid. sulfuric acid and trifluoroacetic acid are particularly preferred.
The reaction temperature usually ranges from -10"C to 100^C (preferably -5 to Although the reaction time differs with the reaction temperature or the like. it ranges from 5 minutes to 48 hours (preferabl\ 30 minutes to 10 hours).
When the hydroxy-protecting group is a "silyl group". deprotection may be conducted by reacting the corresponding compound with a compound containing a fluoride anion. such as tetrabutvlammonium fluoride. in an inert solvent.
There is no particular limitation on the inert solvent used in the above reaction insofar as it is inert to the reaction. Examples include hydrocarbons such as hexane and benzene. halogenated hydrocarbons such as methylene chloride, chloroform and carbon tetrachloride. esters such as ethyl acetate. ketones such as acetone and methyl ethyl ketone. and ethers such as ether. tetrah\drofuran and dioxane: and mixtures thereof with water. Of these. ethers are preferred.
Although there is no particular limitation imposed on the reaction temperature or reaction time, the reaction temperature usually ranges from -10 to 50 0 C (preferably 0 to 30 0 C) and the reaction time usually ranges from 2 to 24 hours (preferably 10 to 18 hours).
Doc FP9907s3.doc P81485/FP-9907 PCT /sa-gad-shEnglish translation of spec (pages 86-134 1 1- 12 00 After completion of the reaction, the desired compound in this reaction is separated from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed. filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate to the filtrate. washing the resulting mixture with water and then distilling off the solvent. If necessary. the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation. column chromatography or the like.
If desired. the hydroxy group of the resulting compound can be esterified or protected.
Esterification of Compound (II) by using 1 to 3 molar equivalents of an esterifying agent can produce a mixture of a compound having 1 to 3 esterified hydroxy groups. By separating the compound from the mixture by column chromatography or the like and then protecting its hydroxy group if desired.
Compound (Ic) is also available.
Doc FP990?s3.doc P8 ]485/FP-c)O7 f PCT tsa-ead-sh'Enlish translation ofspcc (paets 86-134 114 12 00 (Process B) Process B is for the preparation of an ester derivative of Compound By this process. Compound wherein R- is a methyl group. an ester residue is present at the 2'-position. a hydroxy group or ester residue is present at the 2"position and a hydroxy group or ester residue is present at the 3"-position can be prepared.
Process B
OH
O OH
H
HN 0 O R X
C
CONH,
2
O
N NH
O
Step B1 H1O (Ila)
OO
0 O CONH, 2
H
HN N- O _X CH ,O 0 N NH Step B2 (1i1,)
OR-
0
H
HN N 0 O R O0 IXd (Id DR'c
CONH
2
'O
O
N ,NH
O
_H10 OR 3 wherein: R 1 and X have the same meanings as described above. R 3 d represents an ester residue. R 4 b represents a hydrogen atom or an ester residue and Rd represents a hydrogen atom or an ester residue.
Doc FP9907s3.doc P81485!FP-9907( PCT 'isa-gad-hiEnI ish translation of spec (pagcs 86-134 14 12 00(J Step Bi is a step for preparing a compound of formula (liIa). This step is conducted by reacting a compound of formula (IIa) with an acetonide agent in an inert solvent in the presence of an acid catalyst.
Examples of the acetonide agent usable in the above reaction include acetone.
methoxyisopropene and 2.2-dimethoxypropane. of which acetone and 2.2 dimethoxypropane are preferred.
Examples of the acid catalyst usable in the above reaction include inorganic acids such as hydrogen chloride, nitric acid. hydrochloric acid and sulfuric acid.
organic acids such as acetic acid. trifluoroacetic acid. methanesulfonic acid and ptoluenesulfonic acid. Lewis acids such as boron trifluoride and acidic resins such as "Amberlyst 15". of which organic acids and acidic resins are preferred. with ptoluenesulfonic acid and "Amberlyst 15" being more preferred.
The reaction temperature usually ranges from -10 to 100 0 C (preferably 0 to 0 Although the reaction time differs with the reaction temperature and the like.
it usually ranges from 1 hour to 7 days (preferably 10 hours to 3 days).
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed, filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate to the filtrate. washing the resulting mixture with water and then distilling off the solvent. If necessary, the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation. column chromatography or the like.
Step B2 is for the preparation of a compound represented by the formula (Id).
This step is accomplished b\ esterifying Compound (lllal. removing an isopropylidene group from the esterified compound and then esterifving the hvdroxy group if desired.
Esterification is conducted as in the corresponding reaction described in Step A2. while the reaction to remove the isopropylidene group is conducted by reacting the corresponding compound with an acid as in Step BI while using, as an inert solvent. water, an alcohol such as methanol or ethanol or aqueous alcohol.
Doc: FP9907s3.doc P81485/FP-9907(PCT)Itsa-gad-sh/English translaion of spec (pages 86-13441 12 00 (Process C) Process C is for the preparation of an ester derivative of Compound By this process. it is possible to prepare Compound (le) wherein R 2 represents a methyl group. a protected or unprotected hydroxy group or an ester residue is present at the 2"-poisition. and a protected or unprotected hydroxy group or an ester residue is present at the 3"-position.
Process C
OH
0 1 H i HN N.i 0" 0
CONH
2
O
0
O
CH3O OH Step C1 (lib) 0 il H HN i R
"-X
OR%
SCONH
2 0 "0 0,k' 0 oZ.v 0 N. NH i
O
CHO
OR-'
(le i wherein: R 1 and X have the same meanings as described above. represents a hydrogen atom. a hydroxy-protecting group or an ester residue. and represents a hydrogen atom. a hydroxy-protecting group or an ester residue. with the proviso that R'e and R'e represent neither a hydrogen atom nor a hydroxy-protecting group simultaneously.
Step Cl is a step for preparing Compound (Ie) and this step is accomplished by esterifying the compound of the formula (IIb) and. if desired. protecting the hydroxy group.
Esterification is conducted as in the corresponding reaction described in Step A2. A mixture of monoesters may be obtained by the use of an esterifving agent in an Doc FP9907s3.doc P81485/FP-9907(PCT!tsa-gad-sh/English translation of spec (pages 86-134)'14 2 00 amount of about 1 molar equivalent. This mixture can be easily separated by column chromatography or the like. Use of the esterifying agent in an amount of about 2 molar equivalents yields a diester.
The hydroxy-protecting reaction is conducted in a similar manner to that described in Step Al.
(Process D) Process D is for the preparation of an ester derivative of Compound By this process. Compound (If) having a protected or unprotected hydroxy group or an ester residue at the 2"-position. a protected or unprotected hydroxy group or an ester residue at the 3'-position. a protected or unprotected hydroxy group or an ester residue at the 2"-position and a protected or unprotected hydroxy group or an -0ester residue at the 3"-position can be prepared.
Process D
OH
O
O OHCONH 2 O^ 0 HN N O N ,NH R 0 -X HO
OH
Step D1 (Ilc OR H I HN N O R 0
OR'
c
CONH
2
O
",0N NH R -O OR0 R, aO OR 3 wherein: R 1 and X have the same meanings as described above. R'a represents a hydrogen atom. a hydroxy-protecting group or an ester residue. R 3 f represents a hydrogen atom, a hydroxy-protecting group or an ester residue. R 4 c represents a hydrogen atom, a hydroxy-protecting group or an ester residue. and R'r represents a Doc FP9907s3.doc P81485iFP-9907(PCT)!'sa-2ad-sh.'En2lsh translation of spec (paes 86-13441 12 00 hydrogen atom. a hydroxy-protecting group or an ester residue, with the proviso that all of R a. R f. R 4 c and Rr represent neither a hydrogen atom nor a hydroxy-protecting group simultaneously.
Step Dl is a step for the preparation of Compound It can be accomplished by protecting the diol portion of a compound having the formula (IIc) with an isopropylidene group. esterifying the resulting compound. removing the isopropylidene group from the esterified compound and then. esterifying or protecting the hydroxy group if desired.
The protection of the diol portion with an isopropylidene group is conducted in a similar manner to that in Step B1. Use of about 1 molar equivalent yields a mixture of a compound protected at the and 3'-positions and a compound protected at the and 3"-positions. The mixture can easily be separated. for example. by column chromatograph\.
Esterification is conducted in a similar manner to the corresponding reaction in Step A2. Use of an esterifying agent in an amount of about I molar equivalent yields a mixture of monoesters. This mixture can easily be separated. for example. by column chromatography. Use of the esterifying agent in an amount of about 2 molar equivalents yields a diester.
The reaction to remove the isopropylidene group is conducted in a similar manner to the corresponding reaction in Step B2.
The esterification of the resulting compound. which is conducted as desired, is conducted in a similar manner to the corresponding reaction in Step A2. Use of an esterifying agent in an amount of about I molar equivalent yields a mixture of monoesters. This mixture can easily be separated. for example. by column chromatography. Use of the esterifying agent in an amount of about 2 molar equivalents yields a diester. The hydroxy-protecting reaction of the compound thus obtained is conducted in a similar manner to Step Al. Use of a protecting agent in an amount of about 1 molar equivalent yields a mixture of compounds each having one protected hydroxy group. This mixture can easily be separated. for example. by column chromatography. Use of the protecting agent in an amount of about 2 molar equivalents yields a compound having two protected hvdroxy groups.
Compound (If) is also available by esterifying the compound of the formula (IIc) with 1 to 4 molar equivalents of an esterifying agent, separating the resulting Doc. FP9907s3.doc P8] 485,'FP-9907( PCT)i sa-ead-sh/Enslish translafion of spec (pages 86-134)I/4. 12 00 mixture. for example. by- column ch-romatography and if desired. protecting the hydroxy group.
(Process E) Process E is for the preparation of an ether derivative of formula (12) and (1h) of Compound (la).
Doc FP,9907s3.doc DocF~9Os~oc P81485/FP-990 7 1PCT).'tsa-2ad-sh/En2 I sh translation of spec (paes 86-134 1,I4 12 00' Process E 1 H i~ I OH I HN- NI_ IN N -xMeO OH Step Ell I .0 0
CONH
2 HN NH N Step E2 *N 0 o 0, ,N R l x 0M e O O H 0
(IV,,
0- 0 H
CONH-
2 HN- d NO N- 01N
L
0 0 0 -x ~MeO 0 i
(V)
Step E3 HN
N
Ri- 0 C,0 00 MeO 0 RIC Stec) E4
HN
HN 0 UiCONHi 0 0 N .,NH 0 MeO 0 i wherein: R' and X have the same me-aning-s as described above. R' 0 represents the above-described ether residue and L represents a protecting group for the nitrogen atom of the uracil residue.
Doc: FP9907s3.doc Doc:FP907s3doc P148SIFP-9907(PCTI/tsa.2ad-sh/fEngish translation of spec (pages 86-134).14 12 00 Step El is a step for preparing a compound represented by formula (IV) by reacting a compound of formula (IIIa) with an alkylation protecting reagent represented by the formula LY (wherein L and Y have the same meanings as described above) in an inert solvent in the presence of a base.
Examples of the alkylation protecting reagent (LY) usable in the above reaction include 4-methoxybenzyloxymethyl chloride. pivaloyloxymethyl chloride and acetoxymethyl chloride, of which 4-methoxybenzyloxymethyl chloride is preferred.
Examples of the base usable in the above reaction include tertiary amines such as 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1.5-diazabicyclo[4.3.0]non-5-ene (DBN) and alkali metal hydrides such as sodium hydride and potassium hydride. of which 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) is preferred.
Examples of the solvent usable in the above reaction include ethers such as diethvl ether. tetrahydrofuran and dioxane and amides such as N.Ndimethylformamide and N.N-dimethylacetamide. of which N.N-dimethylformamide is preferred.
The reaction temperature usually ranges from -30 to 100°C (preferably -10 to Although the reaction time differs with the reaction temperature and the like, it usually ranges from 30 minutes to I day (preferably I hour to 5 hours).
After completion of the reaction. the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed, filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate. washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate or saturated saline. drying over anhydrous magnesium sulfate or anhydrous sodium sultate and then distilling off the solvent. If necessary.
the resulting product can be purified further in a manner known to those skilled in the art. for example. recrystallization. reprecipitation. column chromatography or the like.
Step E2 is a step for preparing a compound of the formula by reacting a compound of the formula (IV) with an alkylating agent having a desired ether residue in an inert solvent in the presence of a base.
Examples of the alkylating agent usable in the above reaction include alkvl halides and alkyl triflates. of which an alkyl iodide is preferred.
Doc FP9907s3.doc
P
8 1 4 8 5/FP-99071PCT)/sa-gad-sh/English translation of spec (pages 86-134 '14 12 00 Examples of the base usable in the above reaction include tertiary amines such as 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1.5-diazabicvclo[4.3.0]non-5-ene (DBN) and alkali metal hydrides such as sodium hydride and potassium hydride. of which sodium hydride is preferred.
Examples of the solvent usable in the above reaction include ethers such as diethvl ether. tetrahvdrofuran and dioxane and amides such as N.Ndimethvlformamide and N.N-dimethylacetamide. of which N.N-dimethylformamide is preferred.
The reaction temperature usually ranges from -30 to 100°C (preferably 0 to Although the reaction time differs with the reaction temperature and the like.
it usually ranges from 1 hour to 2 days (preferably 1 hour to 10 hours).
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed, filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate, washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate or saturated saline, drying over anhydrous magnesium sulfate or anhydrous sodium sulfate and then distilline off the solvent. If necessary.
the resulting product can be purified further in a manner known to those skilled in the art. for example. recrystallization. reprecipitation. column chromatography or the like.
Step E3 is a step for preparing a compound of the formula (Ig) by reacting a compound of the formula (Vi with an agent capable ofdeprotecting the protected uracil residue in an inert solvent.
When the protecting group contained in the uracil residue in the formula is a 4-methoxybenzyloxymethvy group, examples of the deprotecting agent usable here include 2.3-dichloro-5.6-dicvano- 1.4-benzoquinone (DDQ) or cerium (IV) ammonium nitrate (CAN) [preferably 2.3-dichloro-5.6-dicyano-1.4-benzoquinone while examples of the solvent usable include water, alcohols such as methanol and ethanol. and halogenated hydrocarbons such as methylene chloride and chloroform, and mixtures thereof (preferably a mixed solvent of methylene chloride and water). The reaction temperature usually ranges from 0 to 150°C (preferably to 100 0 Although the reaction time differs with the reaction temperature and the like. it usually ranges from 1 hour to 2 days (preferably 1 hour to 10 hours).
Doc FP9907s3 doc P81485/FP-9907(PCT /tsa-ead-shEnglish translation of spec (pages 86-134 I14 1 2 00 When the protecting group contained in the uracil group in the formula is a pivaloyloxymethyl or acetoxymethyl group. examples of the deprotecting agent usable here include alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, alkali metal carbonates such as sodium carbonate and potassium carbonate. aqueous ammonia. and amines such as methylamine and ethylamine (preferably sodium hydroxide or potassium carbonate). Examples of the solvent include water, alcohols such as methanol and ethanol. ethers such as dioxane and tetrahvdrofuran. and mixtures thereof (preferably a mixed solvent of the alcohols and ethers with water). The reaction temperature usually ranges from 0 to 100°C (preferably 10 to 50 0 Although the reaction time differs with the reaction temperature and the like. it usually ranges from 10 minutes to I day (preferably 1 hour to 10 hours).
After completion of the reaction, the desired compound in the above reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed. filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate. washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate or saturated saline as needed, drvinu over anhydrous magnesium sulfate or anhydrous sodium sulfate and then distilline off the solvent. If necessary. the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation. column chromatography or the like.
Step E4 is a step for preparing a compound of the formula OIh by reacting a compound of the formula (1g vith an acid catalyst in an inert solvent.
Examples of the acid catalyst include inorganic acids such as hydrochloric acid. sulfuric acid and nitric acid. organic acids such as acetic acid. trifluoroacetic acid. trichloroacetic acid. methanesulfonic acid and p-toluenesulfonic acid. Lewis acids such as boron trifluoride and acidic resins such as "Amberlyst 15". of which acetic acid. trifluoroacetic acid. p-toluenesulfonic acid and "Amberlvst 15" are preferred.
Examples of the solvent include water, alcohols such as methanol and ethanol and ethers such as dioxane and tetrahydrofuran. and mixed solvents of the alcohol or ether with water, of which methanol is preferred.
Doc: FP9907s3.doc P81485/FP-9907(PCT)/tsa-gad-shiEnglish translation of spec (pages 86-134 /14.12 00 The reaction temperature usually ranges from 0 to 150 0 C (preferably 10 to 0 Although the reaction time differs with the reaction temperature and the like.
it usually ranges from 1 hour to 2 days (preferably 3 hours to 1 day).
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed. filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate. washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate and saturated saline as needed, and then distilling off the solvent. If necessary. the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation. or column chromatography.
Compound (Ih) thus obtained can be converted to the corresponding h\droxy protected compound. ester derivative or N-alkvlcarbamoyl derivative by any one of Processes A to D and below-described Process F.
Doc FP9907s3.doc P81485.FP-9907(PCT wtsa-gad-sh/Enghlsh translation of spec (pages 86-134.4 4 12 00 117 (Process F.) Process F is for the preparation of an N-alkylcarbamoyl de-rivativ.e of the invention compound (la).
Process F
HN
OH
0--
H
W 0' "'C 0 CONH2 0, N, ,NH 0 MeO OH Step F1 OBz 0 OBz H C0NH 2 N 0 ,N,-NH xMeO OBz Step F2 H N
N
R-0 OB 0 0 3 '0 N NH 0 'Ae C OBz SteD F3
H
H N
N
R-0 '>02 -0 C 0 N NH 0 M e 1 H ,wherein: R' and X have the same meaningps as described above, RI' and R 1 each independently represent the N -alkyl residue of the above-described N-al ky Icarbamoyl group and Bz represents a beazol group.
Doc FP9907s3.doc Do F997s.dc P8 148 5/FP-990 7 PCT)fLsa-gad-shiF ngllsh translat ion of spec (pages 86-134).14 1200 Step Fl is a step for preparing a compound of formula (VI) by reacting a compound of formula (II) with a benzoylating agent in an inert solvent in the presence of a base.
Examples of the benzoylating agent include benzoyl chloride. benzoyl bromide and benzoic anhydride. of which benzoic anhydride is preferred.
Examples of the base usable in the above reaction include organic amines such as triethylamine. 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU). diazabicyclo[4.3.0]non-5-ene (DBN). pyridine and 4-dimethylaminopyridine and alkali metal hvdrides such as sodium hydride and potassium hydride, of which pyridine and 4-dimethylaminopyridine are preferred.
Examples of the solvent usable in the above reaction include ethers such as diethvl ether. tetrahydrofuran and dioxane. amides such as N.N-dimethltformamide and N.N-dimethvlacetamide. halogenated hydrocarbons such as methvlene chloride and chloroform, and pyridine. of which pyridine is preferred.
The reaction temperature usually ranges from -30 to 100'C (preferably -10 to Although the reaction time differs with the reaction temperature and the like.
it usually ranges from 30 minutes to 1 day (preferably 1 hour to 10 hours).
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture if necessary. filterin off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate, washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate and saturated saline as needed. dryinu over anhydrous magnesium sulfate or anhydrous sodium sulfate. and then distilling off the solvent. If necessary, the resulting product can be purified further in a manner known to those skilled in the art. for example. b\ recrystallization. reprecipitation or column chromatography.
Step F2 is a step for preparing a compound of formula (VII) by reacting a compound of formula (VI) with nitrosylsulfuric acid at 0 to 30°C in an inert mixed solvent of methylene chloride and water and then reacting diazomethane with the reaction mixture at 0 to 30 0 C in methylene chloride.
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
Doc FP9907s3.doc P8i4s'/FP-99o7 PCT /tsa-ead-shEnjiish translation of spec (pages 86-134),'14 12 00 The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed. filtering off any insoluble matter, adding a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate, washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate and saturated saline as needed. drying over anhydrous magnesium sulfate or anhydrous sodium sulfate and then distilling off the solvent. If necessary. the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation or column chromatography.
Step F3 is a step for preparing a compound of the formula (Ii) by reacting a compound of the formula (VII) with an amine in an inert solvent.
Examples of the solvent usable in the above reaction include water, alcohols such as methanol and ethanol and amides such as N.N-dimethylformamide and N.Ndimethylacetamide. of which alcohols are preferred.
The reaction temperature usually ranges from 0 to 100°C (preferably 10 to Although the reaction time differs with the reaction temperature and the like.
it usually ranges from 30 minutes to 1 day (preferably 1 hour to 10 hours).
After completion of the reaction, the desired compound in this reaction is recovered from the reaction mixture in a manner known to those skilled in the art.
The desired compound can be obtained, for example. by neutralizing the reaction mixture as needed. filtering off any insoluble matter, addine a water-immiscible organic solvent such as ethyl acetate or methylene chloride to the filtrate, washing the resulting mixture with a diluted aqueous solution of hydrochloric acid. an aqueous solution of sodium bicarbonate and saturated saline as needed. dryin, over anhydrous magnesium sulfate or anhydrous sodium sulfate and then distilling off the solvent. If necessary, the resulting product can be purified further in a manner known to those skilled in the art. for example. by recrystallization. reprecipitation or column chromatography.
Compound (Ii) thus obtained can be convened to the corresponding hydroxy protected compound, ester derivative or ether derivative by using any one of the above-described Processes A to E.
Doc FP9907s3.doc P81485,FP-9907(PCTtsa-ead-sh/Enelish translation of spec (pages 86-1341) 14 12 00 The present invention also provides: Compound A-500359E represented by the following formula (XI): 0
OH
H
3
CO
(XI)
or a salt thereof: Compound A-5003 5QF represented by the follo Ing formula AllI): 0 OH HN HO NH
OH
H
3 C OH
(XII)
Doc FP9907s3.doc DocFP9O~3.ocP81485iFP-99O7(PCT)Itsa-2ad-sh'En~ish trans Iat ion of spec (pages 86-1341/14 12 00 or a salt thereof.
Amide derivative of Compound A-500359F represented by the follo\wing formula (XIII):
HN
(XIII1) or a salt thereof: Compound A-500(JS59H represented by the followitng for-mula (XI\1): 0 OH H OHO N H 0 N
OH
(XIV)
Doc: FP9907s3.doc Doc.FP9O~s3doc P1485/FP-9907 PCT Ptsa-aad-sh/Enelish translation of spec (pages 86-134)114 12 Of) or a salt thereof: Compound A-500359J represented by the following form ula (XV): 0 OH
HN
HO OHO0 NH 2 0 N 01
HO
(XV)
or a salt thereof.
Compound A-500350%1-3 represented by- the followinu formnula (XV'I):
OH
OHO0
NH
HO'
H
3
CO
(XVI)
Doc FP9907s3,doc DocF~qO~sdoc P81 485/'FP-9907 PCT)/tsa-gad-shIEnglish translation of spec (pages 86-134).14 12.00 or a salt thereof: a process for preparing the compound as described in or by cultivating a microorganism capable of producing said compound and belonging to the Streptomyces spp. and recovering the compound from the cultured broth: a process as described in wherein the microorganism belonging to the Streptomyces spp. and capable of producing the compound is Streptomyces griscus SANK60196 (FERM BP-5420) and is capable of producing the compounds as described in or a microorganism which belongs to the Streptomyces spp. and is capable of producing the compound as described in or a microorganism as described in which is Sireptomyvces griscus SANK60196 (FERNI BP-5420): (11) a process for preparing the compound as described in or by cultivating a microorganism (which belongs to the Streptomyces spp. and is capable of producing the compound) by usin.g singly or in combination. S-(2aminoethyl)-L-cysteine. salts thereof and L-allylglycine as an additive to a medium and collecting the compound as described in or from the cultured broth: (12) a composition for the treatment or prevention of infectious diseases which contains the compound as described in or or a pharmacologically acceptable salt thereof as an effective ingredient: (13) use of the compound as described in (1 or or a pharmacologically acceptable salt thereof for the preparation of a medicament for treating or preventing infectious diseases: and (14) a method of treating or preventing infectious diseases, which comprises administering, to a warm-blooded animal. a pharmacologically effecti, e amount of the compound as described in or or a pharmacologically acceptable salt thereof.
Compounds of the present invention represented by any one of the formulae (XII). (XIII). (XIV). (XV) and (XVI) are produced in the culture broth of Strepromyces griseus Strain SANK60196 which belongs to the Streptomyces spp. and has been separated from the soil collected from Mt. Tsukuba/Igaraki-ken; or produced Doc FP9907s3.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 86-1341,14 12 00 1i-24 by microbial conversion in the cultivation process or chemical conversion in the isolation and purification process.
Compound A-500359E of the formula Compound A-500359F of the formula (XII). Amide derivative of Compound A-500359F of the formula (XIII).
Compound A-500359H of the formula (XIV). Compound A-500359J of the formula (XV) and Compound A-500359M-3 of the formula (XVI) of the present invention each contain asymmetric carbons. and each may therefore exist as various optical isomers. In the present invention, isomers of each of Compound A-500359E.
Compound A-500359F. Amide derivative of Compound A-500359F. Compound A- 500359H. Compound A-500359J and Compound A-500359M-3 are represented by the same formula. but the present invention embraces all the isomers including racemic compounds and also mixtures thereof. When a stereospecific synthesis process is adopted or an optically active compound is employed as a starting compound. the isomer of each of Compound A-500359E. Compound A-5000359F.
Amide derivative of Compound A-500359F. Compound A-500359H. Compound A- 500359J and Compound A-500359M-3 may be prepared directly or. if it is prepared in the form of a mixture. each isomer may be obtained in a manner known to those skilled in the art.
Compound A-500359F. Compound A-500359H. Compound A-500359J and Compound A-500359M-3 ofthe present invention can each be converted into the corresponding salt by a method known to those skilled in the art. The present invention embraces such salts of Compound A-500359F. Compound 500359H, Compound A-500359J and Compound A-500359M-3. There is no particular restriction on the nature of the salt of any of Compound A-500359F. Compound A- 500359H. Compound A-500359J and Compound A-500359M-3. provided that it is medically employed and is pharmacologically acceptable. When the salt of Compound A-500359F. Compound A-500359H. Compound A-500359J or Compound A-500359M-3 is employed for the purpose other than a medicament. for example. employed as an intermediate, no limitation is imposed. Preferred examples of such a salt include alkali metal salts such as a sodium salt. a potassium salt. or a lithium salt. alkaline earth metal salts such as a calcium salt or a magnesium salt.
metal salts such as an aluminium salt. an iron salt. a zinc salt. a copper salt, a nickel salt or a cobalt salt. inorganic salts such as an ammonium salt. organic amine salts such as a t-octylamine salt. a dibenzvlamine salt. a morpholine salt. a glucosamine Doc: FP9907s3.doc P81485/FP-9907(PCT)/tsa-gad-shKEnglish Iranslation of spec (pages 86-134)/1 4.12.00 salt. a phenylglycine alkyl ester salt. an ethylenediamine salt. an N-methylglucamine salt. a guanidine salt. a diethylamine salt. a triethylamine salt. a dicvclohexvlamine salt. an N.N'-dibenzylethylenediamine salt. a chloroprocaine salt. a procaine salt. a diethanolamine salt. a N-benzylphenethylamine salt. a piperazine salt and a tetraamethylammonium salt. or a tris(hydroxymethyl)aminomethane salt. and amino acid salts such as a glycine salt. a lysine salt. an arginine salt. an omithine salt. or an asparagine salt. More preferred are salts preferably usable as a pharmacologically acceptable salt such as a sodium salt. a potassium salt and an ammonium salt.
Compound A-500359E. Compound A-500359F. Amide derivative of Compound A-500359F. Compound A-500359H. Compound A-500359J and Compound A-500359M-3 of the present invention and salts thereof may each exist as a solvate. For example. when they are allowed to stand in the air or recrystallized.
water is adsorbed thereto by absorption or a hydrate may be formed. Such a solvate is also embraced in the present invention.
The present invention also embraces all the compounds. so-called prodrugs.
which will be convenrted into Compound A-500359E. Compound A-500359F. Amide derivative of compound A-500359F. Compound A-500359H. Compound A-500359J or Compound A-50035911-3 by metabolism in vivo.
Compound A-500359E. Compound A-500359F. Compound A-500359H.
Compound A-500359J and Compound A-50015911-3 of the present invention which are represented by the formulae XI). (XV) and (XVI) respectively are available by culturing. in a suitable medium. a microorganism belonuine to the SrrepromYces spp. and reco er from the cultured broth. Streptomyces gri.seus Strain SANK 60196 (which will hereinafter be called -Strain SANK60196"). preferred as the microorganism capable of producing Compound A-500359E. Compound A- 500359F. Compound A-500359H. Compound A-500359J and Compound A- 500359M-3 are, as described above. collected and isolated from the soil of Ttsukubasan/Ibaraki Prefecture in a conventional manner. Strain SANK60196 has the biological characteristics as described above.
The various characteristics of the actinomycetes belonging to Streptomyces spp. such as Strain SANK60196 are not stable, but as is well known. they easily change naturally or artificially. The strains usable in the present invention include all such variants. The present invention embraces all the strains belonging to the Doc FP9907s3.doc P81 485/FP-99071 PCT/tsa-gad-sh!English translation of spec (pages 86-134)/14 12 00 Strepiomyces spp. and capable of producing Compound A-500359E. Compound A- 500359F. Compound A-500359H. Compound A-500359J or Compound A-500359M- 3.
Any synthetic or natural medium is usable as a medium for culturing the microorganism capable of producing Compound A-500359E. Compound A-500359F.
Compound A-500359H. Compound A-500359J or Compound A-500359M-3. insofar as it contains a source selected from carbon sources. nitrogen sources. inorganic ions and organic nutrition sources as necessary.
Examples of the nutrition source usable here include known carbon sources.
nitrogen sources and inorganic salts which are conventionally used for the cultivation of a mycotic or actinomycete strain and are utilisable by microorcanisms.
Specific examples of the carbon source include glucose, fructose, maltose.
sucrose. mannitol. glycerol. dextrin. oats. rye. corn starch. potato. corn meal. soybean meal. cotton seed oil. glutinous malt syrup. syrup. soybean oil. citric acid and tartaric acid. They may be used either singly or in combination. The amount of the carbon source to be added usually varies. but is not limited to. within a range of from I to wt.%o of the amount of the medium.
A substance containing a protein or a hydrolysate thereof is generally employed as the nitrogen source. Preferred examples of the nitrogen source include soybean meal. wheat bran. peanut meal. cotton seed meal. skimmed milk. casein hydrolysate. Pharmamine (product of Sheffield Chemical). fish meal. corn steep liquor. peptone. meat extract. pressed yeast. dry yeast. yeast extract. malt extract.
potato. ammonium sulfate. ammonium nitrate and sodium nitrate. It is preferred to use the above-exemplified nitrogen sources either singly or in combination in an amount ranging from 0.2 to 6 0 o of the amount of the medium.
Any ordinarily employed salt containing an ion such as sodium. ammonium.
calcium. phosphate. sulfate. chloride or carbonate can be used as the nutrient inorganic salt. In addition, trace of metals such as potassium. calcium, cobalt.
manganese. iron and magnesium are usable.
The addition of cobalt. skimmed milk or yeast extract is particularly effective in the production of Compound A-500359E. Compound A-500359F. Compound A- 500359H or Compound A-500359J.
Doc FP9907s3.doc P81485/FP-9907(PCT)/tsa-2ad-sh/English translation of spec (pages 86-134)/14 12.00 Upon culturing the microorganism. an inhibitor of antibiotic biosynthesis can be added to produce Compound A-500359E. Compound A-500359F and Compound A-500359H. Compound A-500359E. Compound A-500359F and Compound A- 500359H can each be produced. for example. by using S-(2-aminoethyl)-L-cysteine or salt thereof which is an aspartate kinase inhibitor singly or in combination with cobalt.
skimmed milk and yeast extract. as a medium additive. For example. use of the above-described additive in combination with skimmed milk improves productivity of Compound A-500359E. Compound A-500359F and Compound A-500359H. The additive can be added to give its final concentration ranging from I to 100 mMl. For the production of Compound A-500359E. Compound A-500359F and Compound A- 500359H. the final concentration of 10 mM is preferred.
Use of the above-described additive in combination with an amino acid or salt thereof makes it possible to produce useful compounds related to Compound A- 500359F and Compound A-500359H. In particular. by the use in combination with L-allylglycine or a salt thereof. Compound A-500359M-3 (XVI) is available. The Lallylglycine can be added at a final concentration ranging from 1 to 100 mn.M. At the final concentration of 10 mM. Substance A-500359M-3 can be produced preferably.
Upon liquid culture. an antifoamer such as silicone oil. vegetable oil.
surfactant or the like can be used.
The medium for cultivation of Strain SANK60196 to produce Compound A- 500359E. Compound A-500359F. Compound A-500359H. or Compound A-500359J preferably has a pH ranging from 5.0 to Although the temperature which allow s growth of Strain SANK60196 ranges from 12 to 36"C. the strain is preferably cultured at 18 to 28-C. more preferably 19 to 23°C. in order to produce Compound A-500359E. Compound 500359F. Compound A-500359H and Compound A-500359J.
By in order to obtain Compound A-500359E. Compound A-500359F.
Compound A-500359H. Compound A-500359J and Compound A-500359\M-3. an aerobic culture of Strain SANK60196 can be used. Examples of such a cultivation method include ordinarily employed aerobic culture such as solid culture. shaking culture. and aeration agitation culture.
For small-scale cultivation, shake culture for several days at 19 to 23 0 C is preferred. Cultivation is started by growing a step of seed culture in a first or second Doc: FP9907s3.doc P81485FP-997(PCT /tsa-eadshEne ish translaiion of spec (pages 86-134 I 4 12 00 stage process in a baffled Erlenmeyer flask (equipped with a water flow adjusting wall) or an ordinarily-employed Erlenmeyer flask. A carbon source and a nitrogen source can be used in combination as a medium in the seed culture. The seed culture flask may be shaken at 19 to 23 0 C for 5 days in a thermostat incubator or shaken until the seed culture grows sufficiently. The seed culture thus grown is used for inoculation on the second seed culture medium or a production medium. When the seed cultures are used under an intermediate growing step. they are allowed to grow in a similar manner. followed by partial inoculation into a production medium. The flask into which the seeds have been inoculated is subjected to culturing with shaking at a constant temperature for several days. and after completion of the cultivation, the cultured medium in the flask is centrifuged or filtered.
For large-scale cultivation, on the other hand. use of ajar fermenter or tank equipped with an agitator and an aeration apparatus is preferred. Prior to cultivation in such a container, a nutrient medium is heated to 121 to 130 0 C for sterilization.
After cooling, the seed cultures which have been allowed to grow in advance by the above-described method are inoculated on the sterilized medium. Then. cultivation is carried out with aeration and acitation at 19 to 23 0 C. This method is suitable for preparing a large amount of compounds.
Compound A-500359E. A-500359F or A-500359H can also be produced by adding. as an aspartate kinase inhibitor, an aqueous solution of S-(2-aminoethyl)-Lcysteine or salt thereof which has been previously filter-sterilized in advance to a sterilized medium at the start of. or durin cultivation.
Compound A-500359M-3 can be produced b, separately or simultaneously adding aqueous solutions of 2-aminoethyl -L-cysteine or salt thereof. and L-allyl glycine or salt thereof which have been filter sterilized in advance to the sterilized medium at the start of. or during, cultivation.
The product of Compound A-500359E. A-500359F. A-500359H. A-500359J and A-500359M-3 by cultivation can be measured by subjecting a portion of the cultured broth to HPLC analysis. The titre of Compound A-500359E. A-500359F. A- 500359H. A-500359J and A-500359M-3 usually reaches a peak in 3 to 15 days.
After completion of the cultivation, the cell component is separated from the cultured broth by filtration with the aid of diatomaceous earth or centrifugation and Compound A-500359E. A-500359F. A-500359H. A-500359J and A-500359M-3 Doc FP9907s3 doc P81485/FP-9907(PCT)itsa-gad-sh/Enelish translation ofspec (pages 86-134 i14 12 00 present in the filtrate or supernatant is purified by utilizing their physico-chemical properties with HPLC analytical data as an index. As diatomaceous earth. "Celite 545" (product of Celite Corporation) is preferred. Compound A-500359E. A- 500359F. A-500359H. A-500359J and A-50359M-3 present in the filtrate can be purified by using adsorbents singly or in combination, for example, activated charcoal or an adsorbing resin such as "Amberlite XAD-2 or XAD-4" (product of Rohm Haas). and "Diaion HP-10. HP-20. CHP-20P. HP-50 or SP207" (each. product of Mitsubishi Chemical). Compound A-500359E. A-500359F. A-500359H. A-500359J and A-500359M-3 can be separated from impurities by passing a solution containing Compound A-500359E. A-500359F. A-500359H. A-500359J and A-500359.M-3 through the layer of such an adsorbent as described above, and removing the impurities adsorbed thereto from the solution: or by eluting the adsorbed Compound A-500359E. A-500359F. A-500359H. A-500359J and A-500359M-3 with aqueous methanol. aqueous acetone. aqueous n-butanol. aqueous ammonia. ammoniacontaining aqueous methanol or ammonia-containing aqueous acetone. When an ammonia-containing solution is employed as an eluent. the amide derivative of compound A-500359F happens to be produced upon elution from the column or concentration.
Compound A-500359E. Compound A-500359F. the amide derivative of Compound A-500359F. Compound A-500359H. Compound A-500359.1 and Compound A-500359M-3 thus obtained can be purified by adsorption column chromatography using silica gel. "Florisil". "Cosmosil" (product of Nacalai Tesque).
or "Diaion CHP-20P or SP20'" (product of Mitsubishi Chemical): gel filtration chromatography with "Sephadex G-10 (product of Pharmacia Biotech) or "Toyopearl (product of TOSOH Corporation): anion exchange chromatography with 'Dowex 1 or SBR-P" (product of Dow Chemical) or "Diaion PA316" (product of Mitsubishi Chemical): normal phase and reversed phase HPLC: or the like.
Compound A-500359E. Compound A-500359F. the amide derivative of Compound A-500359F, Compound A-500359H. Compound A-500359J and Compound A-500359M-3 of the present invention can be separated and purified by using the above-exemplified separation and purification means singly or in combination as needed, or in some cases, by using one of them in repetition.
Doc. FP9907s3.doc P81485/FP-9907(PCT /tsa-gad-sh/Enelish translation of spec (pages 86-134 I'14 12.00 Compound A-500359F can be obtained by hydrolysis of Compound A- 500359E. For example. hydrolysis is preferably conducted under basic conditions.
preferably in aqueous basic solution.
Examples of the basic compound usable for hydrolysis include alkali metal hydroxides and weak acid salts thereof such as sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium acetate, sodium carbonate. potassium carbonate and sodium bicarbonate: alkaline earth metal hydroxides and weak acid salts thereof such as calcium hydroxide. magnesium hydroxide and magnesium acetate: inorganic basic compounds and basic salts thereof such as ammonia: organic amines and basic salts thereof such as t-octylamine. dibenzylamine. morpholine. glucosamine.
phenylglycine alkyl ester. ethylenediamine. N-methylglucamine. guanidine.
diethylamine. triethylamine. dicvclohexvlamine. N.N'-dibenzylethylenediamine.
chloroprocaine. procaine. diethanolamine. N-benzylphenethylamine. piperazine.
tetramethvlammonia and tris( hdroxvmethvl )aminomethane. A basic buffer containing an alkali metal ion. an alkaline earth metal ion. an inorganic ion such as ammonia. or an organic amine ion of the above-exemplified basic compounds may also be employed. Among them. alkali metal hydroxides are preferred. of which sodium hydroxide is particularly preferred. In particular, hydrolysis of Compound A- 500359E by using sodium hydroxide can easily produce Compound A-500359F.
The concentration of the basic compound used in the above-described reaction preferably ranges from 0.001 to 1N. more preferably 0.3 to 0.1N. The reaction temperature is preferably -20 to 40'C. more preferably 0 to 30°C. The reaction time is preferably 30 seconds to 15 hours. more preferably 30 minutes to 2 hours.
Use of aqueous ammonia as a base produces the amide derivative of Compound A-500359F together with Compound A-500359F. but these compounds can be separated and purified by the above-described method.
The amide derivative of Compound A-500359F may be produced by reacton of Compound A-500359E with ammonia in a solvent.
Examples of the solvent include water and alcohols such as ethanol and methanol. of which water and methanol are preferred.
Gaseous ammonia may be introduced into the solution of the compound, but a solution of ammonia in water or in an alcohol such as methanol or ethanol is usually used. Preferably, an aqueous or methanolic solution is employed.
Doc FP9907s3.doc P81485/FP-9907 PCT /isa-gad-sh/English translation of spec (pages 86-134)/14 12 00 When aqueous ammonia is employed, its concentration preferably ranges from 0.1 to IN. more preferably 0.3 to 0.7N. The reaction temperature is preferably -20 to 0 C, more preferably 0 to 30 0 C. The reaction time is preferably 30 minutes to hours, more preferably 1 to 4 hours.
When aqueous ammonia is used. in addition to the desired amide derivative of Compound A-500359F. Compound A-500359F is produced by the hydrolysis of the ester. These compounds however can be separated and purified by the abovedescribed methods.
The amide derivative of Compound A-500359F can also be produced by reacting Compound A-500359F with a methylating reagent in a solvent. thereby converting it to the methyl ester derivative, that is. Compound A-500359E. and then reacting the resulting compound with ammonia as described above.
Examples of the methylating reagent include diazomethane and dimethvlsulfuric acid. of which diazomethane is preferred. The methylating reagent for the conversion of Compound A-500359F to Compound A-500359E is preferably added in an amount of 1 to 5 equivalents, preferably 1.5 to 2 equivalents.
Examples of the solvent usable for the above reaction include water and alcohols such as methanol and ethanol. of which water and methanol are preferred.
The reaction temperature is preferably -20 to 40 0 C. more preferably 0 to The reaction time is preferably 30 minutes to 15 hours, more preferably 1 to 2 hours.
After completion of the reaction. Compound A-500359F. Compound A- 500359E. and the amide derivative of Compound A-500359F can be isolated from the reaction mixture by the means selected as needed from those described above in the separation and purification means for Compound A-500359E. Compound A-500359F, the amide derivative of Compound A-500359F. Compound A-500359H. Compound A-500359J and Compound A-50359M-3.
Typical preparation processes for Compound A-500359E. Compound A- 500359F, the amide derivative of Compound A-500359F. Compound A-500359H.
Compound A-500359J and Compound A-50359M-3 are described hereinabove, but preparation processes are not limited thereto and other processes already known to those skilled in the art may also be employed.
Doc: FP9907s3.doc P81485/FP-9907(PCT)/sa-gad-sh/Enghlsh translation of spec (pages 86-134)/14 12 00 Compound A-500359E. Compound A-500359F. the amide derivative of Compound A-500359F. Compound A-500359H. Compound A-500359J and Compound A-500359M-3 of the present invention thus available are novel compounds which have not been described in the literature. Their growth inhibitory activity against general gram positive bacteria or gram negative bacteria can be determined by the disk assay method using normal agar medium (product of Eiken Chemical) or heart infusion agar medium (product of Difco Laboratories). Groth inhibitory activity against .\hcohacteria. gram positive bacteria belongin, to the Actinomyceales. can be determined similarly on the above-described medium added further with glycerin.
Typical evaluation methods of biological activity of Compound A-500359E.
Compound A-500359F. the amide derivative of Compound A-500359F. Compound A-500359H. Compound A-500359J and Compound A-500359M-3 were described so far. but the evaluation method is not limited thereto. but other evaluation methods already known to those skilled in the art can also be employed.
The compounds of the present invention or pharmacologically acceptable salts thereof may be administered through various routes. Examples include oral administration using tablets. capsules. granules. powders. syrups or the like: and parenteral administration using injections (intravenous. intramuscular or subcutaneous), drops. suppositories or the like. These formulations can be prepared in a conventional manner by adding to a medicament ordinarily employed carriers known in the field of pharmaceutical formulation technique such as an excipient.
binder. disintegrator, lubricant. corrigent. adjuvant for solubilization. suspending agent. coating agent and"or the like.
For the formation of tablets. various carriers known conventionally in this field can be employed. Examples include excipients such as lactose. sucrose. sodium chloride, glucose, urea. starch, calcium carbonate. kaolin. crystalline cellulose and silicic acid; binders such as water, ethanol. propanol. simple syrup. glucose solution.
starch solution. gelatin solution. carboxymethyl cellulose, shellac, methyl cellulose.
potassium phosphate and polyvinyl pyrrolidone: disintegrants such as dry starch.
sodium alginate, agar powder. laminaran powder. sodium bicarbonate. calcium carbonate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate. stearic monoglyceride, starch and lactose: disintegration suppressants such as sucrose.
stearin. cacao butter and hydrogenated oil; absorption facilitators such as quaternary Doc FP9907s3.doc P81485/FP-9907(PCT)/Isa-gad-shiEnglish translation of spec (pages 86-134),'14 12 00 133 ammonium salts and sodium lauryl sulfate: humectants such as glycerin and starch: adsorbents such as starch, lactose. kaolin. bentonite and colloidal silicic acid: and lubricants such as purified talc. stearates. boric acid powder and polyethylene glycol.
Tablets can be formed as those having ordinary coating as needed such as sugar coated tablets. gelatin encapsulated tablets. enteric coated tablets, film coated tablets.
or double or multiple layer tablets.
For the formation of pills. various carriers conventionally known in this field can be used. Examples include excipients such as glucose. lactose. cacao butter.
starch. hardened vegetable oil. kaolin and talc: binders such as gum arabic powder.
tragacanth powder. gelatin and ethanol: and disintegrators such as laminaran agar.
For the formation of suppositories. various carriers conventionally known in this field can be employed. Examples include polyethylene glycol. cacao butter.
higher alcohols and esters thereof. gelatin and semi-synthetic glyceride.
For formulation as injections, it is preferred that solutions or suspensions are sterilized and they are made isotonic with the blood. Solutions. emulsions or suspensions can be formed using any diluent conventionally used in this field.
Examples include water. ethanol. propylene glycol. ethoxylated isostearyl alcohol.
polyoxylated isostearyl alcohol and polyoxyethylene sorbitan esters of fatty acid. It is also possible to incorporate, in a pharmaceutical preparation. salt. glucose or glycerin in an amount sufficient for preparing an isotonic solution. or to add an ordinarily employed adjuvant for solubilization. buffer. soothing agent and/or the like.
If necessary, a colourant, preservative, flavor, sweetener or other medicaments may be incorporated.
There is no particular limitation on the content of the compound incorporated as an effective ingredient in the above-described pharmaceutical preparation. It can be chosen suitably from a wide range. In general, it is desired to be contained in an amount of 1 to 70 preferably 1 to 30 wt.% in the whole composition.
There is no particular limitation on the administering method of the abovedescribed pharmaceutical preparation and it is determined depending on the dosage form or age. sex or other conditions of a patient to be administered or seriousness of the disease of the patient. For example, tablets, pills. solutions. suspensions.
emulsions, granules or capsules are administered orally. Injections are administered intravenously either singly or as a mixture with an ordinarily employed fluid replacement such as glucose or amino acid. If necessary, they are singly administered Doc FP9907s3.doc P81485/FP-9907(PCT ltsa-gad-sh/English translation of spec (pages 86-134 14 12.00 intramuscularly, subcutaneously. intracutaneously or intraperitoneally. A suppository is administered rectally.
Although the dose of the pharmaceutical composition differs with the conditions. age and weight of the patient. administration route or dosage form. daily dose usually ranges from 2000 mg (preferably 100 mg) as the upper limit to 0.1 mg (preferably I mg. more preferably 10 mg) as the lower limit per adult. It can be administered once or in several portions a day according to the conditions.
Doc: FP9907s3.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 86-134)/14.12 00 [Best Mode for Carrying out the Invention] The present invention will hereinafter be described more specifically by Examples, Tests and Formulation Examples. It should however be borne in mind that the present invention is not limited to or by them. The process for preparing capuramycin, a known substance. will next be described.
Preparation Example 1: Capuramycin 1) Cultivation ofStrepromyces griseus Strain SANK 60196 (FERM BP-5420) Into each of four 2 L Erlenmeyer flasks (seed flasks). each containing 400 ml of a seed culture medium having the below-described composition. were inoculated four loopfuls of Strain SANK 60196 followed by shaking in a rotary shaker at 28°C and 210 revolutions/min (revolutions per minute: which will hereinafter be abbreviated as Seed culture was thus conducted for 5 days.
Seed culture medium Maltose 30 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g CaCO 3 3 g Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121°C for 30 minutes.
Cultivation was conducted as described below. Described specifically, the seed culture was inoculated at 2°o into each of four 30L jar fermenters. each containing 15 L of a sterilized main culture medium having the below-described composition. followed by cultivation with aeration and agitation at 28 0 C for 8 days.
Main culture medium Glucose 30 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g CoCl 2 -6H 2 0 50 mg CaCO 3 3 mg Antifoamer 50 mg Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 ("CB442": product of NOF Corporation) Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121 °C for 30 minutes 2) Isolation and purification of capuramycin After completion of the cultivation, the cultured broth (52 L) obtained above in I) was filtered with the aid of -Celite 545" (product of Celite Co.) added at 4% The filtrate (50 L) was charged on a "Diaion HP-20" column (product of Mitsubishi Chemical; 12 The resulting column was washed with 18 L of distilled water and the adsorbed substance was eluted with 50 L of 10% aqueous acetone. The eluate was concentrated by "Evapor" to give 15 L of the concentrate.
Upon purification as described later, the active substance of each fraction was monitored by HPLC under the following conditions.
Column: "Senshu Pak ODS-H-2151"" 64 x 150 mm (product of'Senshu Scientific Co.. Ltd.) Solvent: 8% acetonitrile 0.04%, aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 210 nm The resulting concentrate was charged on a "Diaion CHP-20P" column (product of Mitsubishi Chemical: 8 The column was washed successively with 16L each of 10% aqueous methanol and 20% aqueous methanol. followed by stepwise elution of the active substances with 16L of 30% aqueous methanol and 24L of 40% aqueous methanol.
On "Diaion CHP-20P" column chromatography. a peak at a retention time of 17.1 minutes upon the above-described HPLC was mainly detected from a 0 to 8L portion (which will hereinafter be called "Fraction of 30% aqueous methanol eluate; peaks at retention times of 13.7 minutes. 17.1 minutes and 22.6 minutes upon the above-described HPLC were detected from a 8 to 16L portion (which will hereinafter be called "Fraction of 30% aqueous methanol eluate: and a peak at a retention time of 22.6 minutes upon the above-described HPLC was detected from a 0 to 12 portion (which will hereinafter be called "Fraction of the 40% aqueous methanol eluate. These fractions were concentrated by "Evapor", respectively.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/Enelish translation of spec (pages 135-218:Examples)/18.12.00 whereby 8.5 L of Fraction A. 8.5 L of Fraction B and 12.5 L of Fraction C were obtained, each as a concentrate.
A 16 to 24 L portion (which will hereinafter be called "Fraction of the aqueous methanol eluate was concentrated by "Evapor" and lyophilized.
whereby 4.7 g of Fraction D was obtained as a crude powdery product.
Fraction B was charged again on a "Diaion CHP-20P" column (1.5 After washing the column with 3 L of 10% aqueous methanol. the adsorbed material was eluted stepwise with 3L each of 20% aqueous methanol. 30% aqueous methanol and aqueous methanol. From a combined fraction (which will hereinafter be called "Fraction of the 0.5 to 3 L portion of the 20% aqueous methanol eluate and the 0 to 1 L portion of the 30% aqueous methanol eluate. a peak at a retention time of 17.1 minutes in the above-described HPLC was mainly detected: from a combined fraction (which will hereinafter be called "Fraction of the 1 to 3 L portion of the aqueous methanol eluate and the 0 to 0.5 L portion of the 40% aqueous methanol eluate. a peak at a retention time of 13.7 minutes in the above-described HPLC was mainly detected; and from the 0.5 to 3 L portion (which will hereinafter be called "Fraction of the 40% aqueous methanol eluate. a peak at a retention time of 22.6 minutes was mainly detected.
Fraction A was combined with Fraction E (the combined one will hereinafter be called "Fraction while Fraction C was combined with Fraction G (the combined one will hereinafter be called "Fraction Fractions F. H and I were concentrated on "Evapor" and lyophilized. respectively. whereby 16.2 g of Fraction H. 33.6 g of Fraction I and 8.6 g of Fraction F were obtained, each as a crude powdery product.
The resulting crude powdery product of Fraction H (16.2 g) was dissolved in 250 ml ofdeionised water. The resulting solution was charged on a "Toyopearl HWcolumn (product of TOSOH Corporation: 4 followed by development with deionised water. As a result of fractionation of the eluate to 75 ml portions each. the active substance having a retention time of 17.1 minutes in the above-described HPLC was eluted in Fraction Nos. 41 to 63. These fractions were collected and concentrated by "Evapor" into 820 ml and the resulting concentrate was lyophilized to give 6.4 g of a crude powdery product.
The crude powdery product thus obtained was dissolved in 400 ml of water.
Each of the 80 ml portions of the resulting solution was charged on an HPLC column Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/1 8.12.00 (YMC-Pack ODS R-3105-20 (1004 x 500 mm: product of YMC Co., Ltd.)) equilibrated with a 6% aqueous solution of acetonitrile, followed by column development at a flow rate of 200 ml/min. The ultraviolet absorption of the active substance at 210 nm was detected and a peak eluted at a retention time of 105 to 120 minutes was collected by five fractionation. each in portions of 400 ml.
The resulting fractions were combined and concentrated by "Evapor" into 330 ml. followed by lyophilization. whereby 3.6 g of a substance was obtained in pure form. The substance was identified as capuramycin. a known antibiotic, by structural analysis.
Example 1: Preparation ofA-500359A (Exemplification (exemp.) Compound No. 1) The crude powdery product (33.6 g) of Fraction I obtained in Preparation Example 1 was dissolved in 450 ml ofdeionised water. The resulting solution was charged on a "Toyopearl HW-40F" column (8 followed by elution with deionised water. As a result of fractionation of the eluate into 150 ml portions, the active substance exhibiting a retention time of 22.6 minutes in HPLC was eluted in Fractions Nos. 47 to 73. These fractions were collected. concentrated by "Evapor" into 1.5 L and then lyophilized to give 25 g of a crude powdery product.
The resulting crude powdery product (25 g) was dissolved in 300 ml of deionised water. The resulting solution was charged on a "Cosmosil 140C18-OPN" column (product of Nacalai Tesque: 1.5 After washing the column with 3 L of deionised water and 12 L of 1% aqueous acetonitrile. the active compound was eluted with 6 L of 10% aqueous acetonitrile. The eluate was concentrated by "Evapor" into 840 ml and insoluble matter was filtered from the concentrate. The filtrate was lyophilized to give 20 g of Substance A-500359A in pure form. The following data are physico-chemical properties of the resulting substance.
I) Appearance of the substance: white powder 2) Solubility: soluble in water and methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C 14
H
33
N;OI
2 4) Molecular weight: 583 (measured by FAB mass spectrometry) Accurate mass. as measured by high-resolution FAB mass spectrometry is as follows: Doc: FP9907s4.doc P81 4 85/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Found: 584.2189 Calculated: 584.2205 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 257nm 10,300) 7) Optical rotation: optical rotation measured in methanol exhibits the following value: [aD 2 0 +94.7" (c 1.00. MeOH) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following maximum absorption: 3380. 2940. 1690. 1520. 1460. 1430. 1390. 1270. 1110. 1060cm'.
9) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard. 'H nuclear magnetic resonance spectrum is as follows: 1.22(3H.d.J=6.7Hz), 1.29(1H.m). 1.49(1H.m). 1.78(1H.m). 1.87(1H.m). 1.92(1H.m), 2.01(1H.m), 3.44(3H.s), 3.58(1H.m). 3.86(1H.br.t.J=4.6Hz).
3.96 (1H.ddd,J=0.7,4.5.5.7Hz). 4.30(1H.t.J=5.2Hz). 4.37(1H.t,J=4.1 Hz).
4.56(1H.dd.J=2.0,11.9Hz). 4.58(1 H.dd.J=2.0.4.3Hz). 4.67(1H.d.J=2.0Hz).
5.23(1H.d.J=5.8Hz). 5.72(1H.d.J=8.1Hz). 5.88(1H.d.J=5.2Hz).
6.02(1 H.br.dd.J=0.7.3.9Hz). 7.91 (1H.d.J=8.1 Hz) ppm.
13C nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard. "C nuclear magnetic resonance spectrum is as follows: 2 28.4(t), 32.1(t), 37.9(t). 50.1(d). 53.5(d). 58.8(q). 63.6(d). 68.8(d). 74.6(d).
79.2(d). 81.1(d). 83.6(d). 90.4(d). 101.3(d). 102.9(d). 109.3(d). 142.0(d). 144.4(s).
152.4(s), 161.9(s), 166.1(s). 173.5(s). 175.3(s) ppm.
11) High performance liquid chromatography Column: "Senshu Pak ODS-H-2151". 6 x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 8% acetonitrile water Flow rate: 1.0 ml/min Detection: UV 210 nm Retention time: 20 minutes.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/1 8.12.00 Example 2: Preparation of A-500359C (Exemp. compound No. 2) The crude powdery product (8.6 g) of Fraction F was dissolved in 500 ml of deionised water. The resulting solution was charged on a "Toyopearl column (8.5 which was developed with deionised water. As a result of fractionation of the eluate into 150 ml portions. the active substance exhibiting a retention time of 13.7 minutes in HPLC was eluted in Fraction Nos. 44 to 82. These fractions were collected, concentrated by "Evapor" into 900 ml. and lyophilized.
whereby 2.2 g of a crude powdery product was obtained.
The resulting crude powdery product (2.2 g) was dissolved in 150 ml of deionised water. The resulting solution was charged on a "Cosmosil 140C I8-OPN" column (product of Nacalai Tesque: 1.5 After washing the column successively with 3 L of deionised water. 3 L of 0.5% aqueous acetonitrile. 3 L of 1% aqueous acetonitrile and 15 L of 2% aqueous acetonitrile. the active substance was eluted with of 4% aqueous acetonitrile. The fraction was concentrated by "Evapor" into 500 ml and then lyophilized. whereby 550 g of a crude powdery product was obtained.
The crude powdery product was dissolved in 80 ml of deionised water. The resulting solution was charged on an HPLC column (YMC-Pack ODS R-3105-20 (1000 x 500 mm: product of YMC)) equilibrated with a 6% aqueous solution of acetonitrile. and the column was developed at a flow rate of 200 ml/min. The ultraviolet absorption of the active fraction at 210 nm was detected and the active fraction eluted at a retention time of from 167 to 180 minutes was collected by fractionation.
The resulting fraction was concentrated into 50 ml by "Evapor". followed by lyophilization. whereby 210 mg of Compound A-500359C was obtained in pure form.
The following data are physico-chemical properties of the resulting substance.
1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C2 3
H
3 1 Ns0 1 4) Molecular weight: 569 (as measured by FAB mass spectrometry) Accurate mass, as measured by high-resolution FAB spectrometry is as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 Found: 570.2034 Calculated: 570.2049 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 257 nm (e 10,700) 7) Optical rotation: optical rotation measured in water exhibits the following value: +890 (c 0.44. HO0) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3390. 2930, 1690. 1520. 1460. 1430. 1390. 1270. 1110. 1060 cm'.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 1.20(3H.d.J=6.7Hz). 1.29(1H.m). 1.62(1H.m). 1.72(1H.m). 1.75(lH.m). 1.90(1H.m).
1.92(1H.m). 3.65(1H.m). 4.11(1 H.dd.J=5.2.6.3Hz). 4.15(1 H.ddd.J= 1.4.4.2.4.3Hz).
4.18(1 H.dd.J=3.3.5.2Hz). 4.43(1 H.dd.J=2.1.6.3Hz). 4.49(1 H.dd.J=3.0.4.4Hz).
4.62(1 H.dd.J=1.7,10.8Hz). 4.76(1H.dJ=2.1 Hz). 5.36(1H.d.J=4.0Hz).
5.77(1H.d.J=3.3Hz). 5.84(1H.d.J=8.1 Hz). 5.98(1H.br.dd.J=1.3.3.0Hz).
7.72(1H.d.J=8.1Hz) ppm.
'1C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1.4-dioxane (67.4 ppm) as an internal standard. 'C nuclear magnetic resonance spectrum is as follows: 21.0(q). 26.8(t). 29.4(t). 35.4(t). 48.9(d). 52.6(d). 61.9(d). 65.3(d). 69.4(d). 73.8(d).
76.7(d). 83.1(d). 89.7(d). 100.1(d). 101.9(d). 109.1(d). 141.0(d). 141.8(s). 151.6(s).
161.7(s). 166.4(s), 173.5(s). 175.8(s) ppm.
11) High performance liquid chromatography Column: "Senshu Pak ODS-H-2151". 66 x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 8% acetonitrile water Flow rate: 1.0 ml/min Detection: UV 210 nm Retention time: 13 minutes.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218.Examples)/l 8.12.00 Example 3: Preparation of A-500359D (Exemp. compound No. 3) An 800 mg portion of the crude powdery product obtained as Fraction D was dissolved in 10 ml of deionised water. A 500 pl portion of the resulting solution was charged on an HPLC column ("Senshu Pak Pegasil ODS (204 x 250 mm. product of Senshu Scientific)) which had been equilibrated with a developing solvent containing acetonitrile. methanol and 0.04% aqueous trifluoroacetic acid at 3:21:76. and the column was developed with the same solvent at a rate of 9 ml/min. The ultraviolet absorption of the active fraction at 210 nm was detected and a peak eluted during to 38 minutes was collected by fractionation. The procedure was carried out 20 times to elute the (in portions of 10 ml).
The powder (15 mg) obtained by concentrating the fractions eluted during to 38 minutes and lyophilizing the concentrate was chromatographed again on the same HPLC column and then. concentrated and lyophilized. whereby 7 mg of Compound A-500359D was obtained in pure form.
The following data are the physico-chemical properties of the resulting substance.
1) Appearance of the substance: white powder 2) Solubility: soluble in water and methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C2 4 H-;NsO I 4) Molecular weight: 567 (as measured by FAB mass spectrometry) Precise mass. as measured by high-resolution FAB mass spectrometry is as follows: Found: 568.2239 Calculated: 568.2254 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 244 nm (e 10.000) 7) Optical rotation: optical rotation measured in water exhibits the following value: [oc]D 2 0 +680 (c 0.69. HO) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3397, 2925, 1683, 1514, 1461. 1432. 1385, 1265, 1205, 1095, 1061 cm'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 1.12(3Hd,J=8. 1Hz), 1.17(1 1.40(1 1.67(1H.m). 1.80(1 1.88(1 H.m).
1.90(1Hm), 2.33(1 3.24(3H.s), 3.50(1 3.57(1 H.t.J=4.7Hz).
4.08(1 H,t,J=4.8Hz), 4.37(m),4.40(m). 4.46(1 H.br.d.J=10.7Hz). 4.50(1H.d.J=2.0Hz).
5.30(1Hbr.s), 5.64(1HdJ=8. 1Hz), 5.73(1H.d.J=4.8Hz). 5.97(1H.d.J=2.4Hz).
7.77(1H.dJ=8. 1Hz) ppm.
13C nuclear magnetic resonance spectrum was measured in deuterated methanol with the signal of methanol as 49.15 ppm. 'C nuclear magnetic resonance spectrum is as follows: 22.3(q). 28.6(t). 32.3(t). 35.8(t). 38.0(t). 50.2(d). 53.6(d). 58.8(q). 60.7(d). 74.7(d).
77.7(d). 80.9(d), 83.8(d). 90.7(d). 99.5(d). 103.0(d). 112.3(d). 142.0(d). 144.1(d).
152.4(s). 162.4(s). 166.3(s). 173.6(s). 175.5(s) ppm.
11) High performance liquid chromatography Column: "Cosmosil 5Cl8-MS". 4.64 x 150 mm (product of Nacalai Tesque) Solvent: a 3:21:76 mixture of acetonitrile methanol 0.04% aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 210 nm Retention time: 9.2 minutes.
Example 4: Cultivation of SrepronmYces griseus Strain SANK 60196 (FERM BP- 5420) Into each of three 2L Erlenmeyer flasks (seed flasks) each containing 500 ml of a medium having the below-described composition were inoculated. in a sterile condition, four loopfuls of Strain SANK60196. followed by shaking in a rotary shaker at 23 0 C and 210 rpm. Seed culture was thus conducted for 5 days.
Seed culture medium Maltose 30 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g CaCO 3 3 g Doc: FP9907s4.doc P8 1485/FP-9907(PCT)/tsa-gad-shlEnglish translation of spec (pages 135-21 8:Examples)/18.12.00 Antifoamer 50 mg (CB442) Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121 C for 30 minutes Cultivation was conducted as described below. Described specifically, the seed culture was inoculated at 3% into each of two 30 L jar fermenters. each containing 15 L of a sterilized medium having the below-described composition. On Day 1 after the commencement of cultivation at 23°C. filter sterilized S-(2aminoethyl)-L-cysteine hydrochloride was added to give a final concentration of 8 mM. and cultivation was then carried out with aeration and agitation for 7 days.
Cultivation medium Maltose Yeast extract 5 g (product of Difco Laboratories) Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Cobalt chloride hexahydrate 0.5 g CaCO 3 3 g Antifoamer 50 mg (CB442) Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121 0 C for 30 minutes Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 Example 5: Preparation of A-500359G (Exemp. compound No. After completion of the cultivation, the cultured broth (28 L) obtained in Example 4 was filtered with the aid of"Celite 545".
Upon purification as described later, the active fraction was monitored by the following high performance liquid chromatography (HPLC).
Column: "Senshu Pak ODS-H-2151 64 x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 8% acetonitrile 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 ml/min Detection: UV 210 nm Retention time: 4.6 minutes 37 L of the resulting filtrate was charged on a "Diaion HP-20" column (5.5 L).
After washing the column with 11 L of deionised water, the adsorbed substance was eluted with 11 L of 10% aqueous acetone. The eluate was concentrated to remove acetone. The residue was lyophilized. whereby 40 g of a crude powdery product was obtained.
The resulting crude powdery product was dissolved in 1 L of distilled water and charged on a "Diaion CHP-20P" column (3 The column was then washed with 6 L of distilled water, and the adsorbed substance was eluted successively with 6 L of each of 5% aqueous methanol. 10% aqueous methanol and 15% aqueous methanol. The 15% aqueous methanol eluate was concentrated to remove methanol.
The residue was lyophilized to give 1.27 g of a powder.
The resulting powder was dissolved in 30 ml of distilled water and the resulting solution was charged on a "Toyopearl HW40F" column (500 ml). followed by elution of the column with distilled water. The eluate was collected by fractionation in portions of 10 ml. each. The active substance having a retention time of 4.6 minutes in the above-described HPLC was eluted in fractions Nos. 41 to 46.
The resulting fractions were concentrated and lyophilized to give 134 mg of a powder.
The resulting powder was dissolved in 3 ml of water and a 750 pl portion of the resulting solution was charged on an HPLC column ("Senshu Pak ODS-H-5251" mm x 250 mm; product of Senshu Scientific)) equilibrated with 4% aqueous acetonitrile containing 0.04% of aqueous trifluoroacetic acid. The column was developed at a flow rate of 10 ml/min. The ultraviolet absorption of the active Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 substance of 210 nm was detected and a peak eluted during 27 to 30 minutes was collected by fractionation. The process was carried out four times.
These fractions eluted during 27 to 30 minutes were concentrated and lyophilized to afford 20 mg of a powder. The resulting powder was dissolved in 1.6 ml of water and a 800 pl portion of the resulting solution was charged on the abovedescribed HPLC column using instead, as a developing solvent, a 5% aqueous acetonitrile solution containing 0.04% of TFA. The column was developed at a rate of 10 ml/min. The active substance showing ultraviolet absorption at 210 nm was detected and a peak eluted during 19 to 20 minutes was collected again by fractionation. The fractions were concentrated and lyophilized. wherebv 14 mg of Compound A-500359G was obtained in pure form. The substance has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water, slightly soluble in methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C-2HQNsO 4) Molecular weight: 555 (as measured by FAB mass spectrometry) Accurate mass. as measured by high-resolution FAB mass spectrometry is as follows: Found: 556.1891 Calculated: 556.1890 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 257 nm (e 10.000) 7) Optical rotation: optical rotation measured in water exhibits the following value: [a]D 2 0 +1090 (c 0.72. HO) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3367. 2931, 1684. 1518. 1482. 1464. 1436. 1408. 1385. 1335. 1272. 1205. 1177.
1114, 1063 cm'.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: Doc: FP9907s4.doc P81 4 85/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 1.37 (1IH.mi). 1.65 (1 H. in). 1.71 (1IH. ml. 1.79 (1IH.mT). 1.92 (1 H.mi). 1.98 (1 H. in).
3.29 (1IH. mn). 3.36 (1 H. in). 4.10 (1 H. dd. J=5.0. 6.5 Hz). 4.14 01H. dt. J=1.5. 4.4 Hz), 4.17 (1IH. dd. J=3.2. 5.0 Hz). 4.41 (1 H. dd. J=2.1. 6.5 Hz). 4.47 (1 H. dd. J=2.9. 4.4 Hz). 4.61 (1 H. dd. J=1.8. 11.4 Hz). 4.78 (1IH). 5.35 (11H. d. J=4.1 Hz). 5.75 d.
J=3.2 Hz). 5.82 (1IH. d. J=8.2 Hz). 5.97 (1IH. dd. J=1.5. 2.9 Hz). 7.71 (1IH. d. J=8.2 Hz) ppm.
13 C nuclear magnetic resonance spectrum was measured in deuterium oxide with I .4-dioxane (67.4 ppm) as an internial standard. 'S3C nuclear magnetic resonance spectrum is as follows: 28.2 28.4 30.5 42.2 533(d). 61.7 66.1 70.2 74.5 77.5 83.9 90.5 100.9 102.7 109.9 141.8 142.7 152.2 162.6 166.9 174.3 177.6 ppm.
I11) High performance liquid chromatography Column: "'Senshu Pak ODS-H-2115 1 66 x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 8% acetonitrile 0.04%, aqueous trifluoroacetic acid Flow rate: 1.5 mrl/Min Detection: UV 210 nm Retention time: 4.6 minutes Example 6: Cultivation of Strc'plolnLcs ('risllus Strain SANK6O 196 (FERM BP- 5420) Into each of four 2 L Erlenmnever flasks (seed flasks) each containinu 500 ml of a medium haviniz the belowk-described composition were inoculated, in a sterile condition. four loopfuls of Strain SANK601I96. and cultivation was then carried out with shaking in a rotary shaker at 23"C and 210 rpm. Seed culture was thus conducted for 3 days.
Seed culture medium Maltose 30 L, Meat extract 52 Polypeptone 5 a Sodium chloride 5 g CaCO 3 3g Doc: FP9907s4.doc Doc: P99074.doc P81 485/FP.9907(PCT)/tsa-,gad-shEnglish translation of spec (pages 135-2 18: Examples Il 8,12.00 Antifoamer 50 mg (CB442) Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121 °C for 30 minutes The culture was conducted as described below. Described specifically, the seed culture broth was inoculated at 3% into each of two 30 L jar fermenters.
each containing 15 L of a sterilized medium having the below-described composition.
Six hours after commencement of cultivation at 23°C. filter-sterilized S-(2aminoethyl)-L-cysteine hydrochloride was added to give a final concentration of mM. and cultivation with aeration and agitation was then carried out for 6 days.
Cultivation medium Maltose 30 g Yeast extract 5 g (product of Difco Laboratories) Meat extract 5 Polypeptone 5 g Sodium chloride 5 g CaCO 3 3 g Antifoamer 50 mg ("CB442") Tap water 1000 ml pH before sterilization: 7.4 Sterilization: at 121 C for 30 minutes Example 7: Preparation of A-500359 M-2 (Exemp. compound No. 396) After completion of cultivation, the cultured broth (30 L) obtained in Example 6 was filtered with the aid of"Celite 545".
Upon purification as described later, the active fraction was monitored by the following high performance liquid chromatography (HPLC) method.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 149 Column: "Senshu Pak ODS-H-2151" 6) x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 8% acetonitrile 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 ml/min Detection: UV 210 nm Retention time: 13.6 minutes L of the resulting filtrate was charged on a "Diaion HP-20" column (6 L).
After washing the column with 12 L of deionised water, the adsorbed substance was eluted with 10% aqueous acetone. The fraction eluted in 12 to 24 L was concentrated to remove acetone. The residue was lyophilized, whereby 12 g of a crude powdery product was obtained.
The resulting crude powdery product was dissolved in 650 ml of distilled water.
The resulting solution was charged on a "Diaion CHP-20P" column (1 The column was then washed with 2 L of distilled water, and the adsorbed substance was eluted with 2 L of 20% aqueous methanol and 4 L of 30% aqueous methanol. The 2 to 4 L portion of the 30% aqueous methanol eluate was concentrated to remove methanol. The residue was lyophilized to yield 2.8 g of a powder.
The resulting powder was dissolved in 50 ml of distilled water and the resulting solution was charged on a "Toyopearl HW40F" column (500 ml), followed by development of the column with distilled water. The eluate was fractionated in portions of 12 ml, each. The active substance having a retention time of 13.6 minutes in the above-described HPLC was eluted in Fraction Nos. 40 to 47. The resulting fractions were concentrated and lyophilized to give 841 mg of a powder.
The resulting powder was dissolved in 23 ml of water and a 1 ml portion of the resulting solution was charged on an HPLC column ("Senshu Pak ODS-H-5251" (20 mm x 250 mm; product of Senshu Scientific)) equilibrated with an aqueous solution containing 0.04% trifluoroacetic acid, 4% acetonitrile and 10% methanol. The column .was developed at a flow rate of 10 ml/min. The ultraviolet absorption of the active substance of 210 nm was detected and a peak eluted during 23 to 26 minutes was collected by fractionation, the preparation being carried out 23 times.
The fractions eluted during 23 to 26 minutes were concentrated and lyophilized to afford 421 mg of a powder. The resulting powder was dissolved in 40 ml of water again and the resulting solution was charged on the above-described Doc: FP9907al.doc PS145/FP-9907(PCTYa/gad/shlcorrected pages of spec/03/01/01 HPLC column using instead, a 7% aqueous acetonitrile solution containing 0.04% of TFA as a developing solvent. The column was developed at a rate of 10 ml/min. The ultraviolet absorption of the active substance of 210 nm was detected and a peak eluted during 33 to 35 minutes was collected again by fractionation. the process being carried out in 40 times. The fractions were concentrated and lyophilized. whereby 190 mg of Substance A-500359 M-2 was obtained in pure form.
The substance has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water and methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C 23
HI
3
N
5 0i 2
S
4) Molecular weight: 601 (as measured by FAB mass spectrometry) Accurate mass, as measured by high-resolution FAB mass spectrometry is as follows: Found: 602.1779 Calculated: 602.1769 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 244 nm (e 14.000) 7) Optical rotation: optical rotation measured in water exhibits the following value: 58 (c 0.39. H-O) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3390. 2937. 1683. 1510. 1461. 1432. 1411. 1344. 1268. 1206, 1179. 1135. 1071. 1023
I
cm.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 1.30(3H.d.J=6.8Hz). 2.63(2H.m). 2.76(1H.dd.J=2.9.14.4Hz).
2.84(1H.dd.J=8.8,14.4Hz). 3.28(3H.s). 3.73(1H.dd.J=5.0.6.5Hz). 3.98(1 H.m).
4.9(1 H.ddd.J 1.5,3.5,4.4Hz). 4.38(1 H.dd.J=3.2.5.0Hz). 4.47(1H.dd.J=2.6.6.5Hz), 4.50(1H,dd.2.6,4.4Hz). 4.73(1H.d.J=2.6Hz), 5.02(1H,dd.J=2.9,8.8Hz).
5.39(1H.d.J=3.5Hz), 5.75(1H.d.J=3.2Hz), 5.85(1H,d,J=8.1Hz), 6.03(1H,dd,J=1.5.2.6Hz), 7.74(1H.d,J=8.1Hz) ppm.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 13C nuclear magnetic resonance spectrum was measured in deuterium oxide Aith 1.4-dioxane (67.4 ppm) as an internal standard. 1 3 C nuclear magnetic resonance spectrum is as follows: 2 1.3 30.0(t), 36.3(t).53.2(d). 55.9(d), 58.6(q), 62.7(d). 65.7(d). 72.7(d). 76.5(d).
78.9(d), 82.4(d), 91.1(d), 100.3(d). 102.7(d). 110.6(d). 141.9(d). 142-3(s). 152.1(s).
162.3(s), 166.9(s), 173 174.5(s) ppm.
11) High performance liquid chromatography Column: "Senshu Pak ODS-H-215 F, 6 x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 8% acetonitrile 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 mI/mml Detection: UJV 2 10 rin Retention time: 14.4 minutes In the below-described Examples. Me. TBS. THE. TBAF. DMIAP and WSC stand for a methyl group. a tert-butyldimethylsilyl group. tetrahydrofuran.
tetrabutyl ammonium fluoride. 4-(dimethy lamino )pvridine and I1-ethy1-3)-(3di methyl aminopropyl)carbodiimide hydrochloride. respectively.
Doc: FP9907s4.doc Doc:FP907s4doc P 1485/FP-9907( PCT)/rsa-gad-sh/English translation of spec (pages 13 5-21 8: Examples)/ 18.12. 00 152 Example 8 (Exemp. compound No. 135) 0 O OH 0 0 H H N N NH
CH
3 0 OH (8-1) Capuramycin (2 g) was dried by azeotropy twice with pyridine and dissolved in 34 mL of pyridine. To the resulting solution, 1.59 g of tert-butyldimethylsilyl chloride was added, followed by stirring at room temperature. Three days later, the solvent was distilled off under reduced pressure. The residue was dissolved in 200 mL of ethyl acetate. The resulting solution was washed with 200 mL of saturated saline and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was charged on a silica gel column (300 which was developed with methylene chloride methanol (concentration gradient from 97:3 to 90:10. which will hereinafter be described as "97:3 to 90:10").
whereby 474.6 mg of the below-described compound was obtained.
OH
o a O 0 H 'OCONH 2 HN N 0 N NH j 0 0
CH
3 O OTBS 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 1H nuclear magnetic resonance spectrum is as follows: 6 7.99 J 8.1 Hz. 1H). 6.02 J 3.7 Hz. 1H). 5.88 J 5.1 Hz. 1H). 5.74 J 8.1 Hz. 1H). 5.23 J 5.8 Hz. 1H). 4.69 1H). 4.61 J 2.2 Hz. 1H).
4.51 J 11 Hz, 1H), 4.41 J 4.7 Hz. 1H). 4.36 J 4.6 Hz. 1H). 3.90 (m.
1H). 3.85 1H). 3.47 3H). 3.30-3.20 2H). 2.02 2H). 1.84 2H). 1.54- 1.28 2H), 0.86 9H). 0.05 6H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3368, 2931, 2858, 1687, 1510. 1473, 1463, 1436, 1385, 1334, 1266, 1145. 1101. 1064 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-shlEnglish translation of spec (pages 135-218:Examples)/l 8.12.00 (8-2) In 3 mL of pyridine were dissolved 100 mg of the compound obtained in (8-1) and 2 mg of DMAP. To the resulting solution was added 145 mg of palmitic anhydride, followed by stirring at room temperature. Forty minutes later, the solvent was distilled off under reduced pressure, and the residue dissolved in 20 mL of ethyl acetate. The resulting solution was washed with 20 mL of saturated aqueous sodium bicarbonate and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was charged on a silica gel column (14 which was developed with methylene chloride methanol (98:2 to 95:5).
whereby 42.7 mg of the following compound was obtained.
0
O
,OH 0 0 H OCONH 2 0 NO 0 N NH 0 0
CH
3 0' OTBS 1) H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 9.17 (br s. 1H), 7.88 2H). 7.47 (br s. 1H). 6.58 (br s, 1H). 6.04 2H). 5.78 2H). 5.58 1H). 5.12 J 7.7 Hz. 1H). 4.64 1H). 4.60 IH). 4.50 (m, 2H). 4.06 1H). 3.88 1H). 3.46 3H). 3.27 3H). 2.37 2H). 2.16-1.10 32H). 0.88 12H). 0.06 6H) ppm.
(8-3) In 53 4L of THF were dissolved 41 mg of the compound obtained in A 53 pL THF solution containing 1M ofTBAF was added to the resulting solution and the mixture stirred at room temperature. Four hours later, the solvent was distilled off under reduced pressure. The residue was charged on a silica gel column (6 which was developed with methylene chloride methanol (96:4 to 94:6). whereby 16.3 mg of the below-described compound was obtained as a desired compound of Example 8.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 0
O
0 H OHCONH 2 rY° HN N N NH
OH
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 =7.76 J 8.1 Hz. 1H). 5.88 J 3.7 Hz. 1H). 5.79 J 5.1 Hz. 1H). 5.72 (d.
J 8.1 Hz. 1H). 5.42 1H). 5.21 J 4.7 Hz. 1H). 4.61 J 2.2 Hz. 1H). 4.54- 4.46 2H). 4.17 2H). 3.71 J 4.8 Hz. 1H). 3.32 3H). 3.18 2H). 2.33 J 7.3 Hz, 2H), 1.98-0.79 35H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3379. 2925. 2855. 1690. 1507. 1462. 1384. 1334. 1262. 1115 cm'.
Example 9 (Exemp. compound No. 280) o 0
OO
0 H CONH 2 0 HN 0 N NH 0 .0 0
CH
3 d bOH (9-1) In 4.5 mL of pyridine were dissolved 150 mg of the compound obtained in Example 69 pL of heptanoic anhydride and 3 mg of DMAP. In a similar manner to that described in Example the resulting solution was reacted.
whereby 286 mg of the following compound was obtained.
O O 0 0 0 H CONH 0 HN N O N YNH
CH
3 d bTBS Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 (9-2) In 250 pL of THF was dissolved 286 mg of the compound obtained in Example To the resulting solution was added 250 L of a THF solution containing IM of TBAF. The resulting mixture was reacted in a similar manner to that described in Example whereby 96.3 mg of the below-described compound was obtained as the desired compound of Example 9.
0 0 0 CONH 2
O
HN N N NH 0 0
CH
3 0 bH 1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.72 J 8.1 Hz. 1H). 5.99 1H). 5.87 J 8.1 Hz. 1H), 5.81 J 4.6 Hz. 1H). 5.72 1H). 5.63 1H). 5.45 J 3.2 Hz. 1H), 4.68 J 2.2 Hz.
1H). 4.59 1H). 4.46 1H). 4.18 J 4.8 Hz. 1H). 3.65 J 5.1 Hz. 1H).
3.34 3H). 3.25 2H). 2.40-2.25 4H). 2.03 2H). 1.85 2H). 1.70-1.50 6H). 1.45-1.25 12H). 0.90 6H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3342. 29 1.2859.1748. 1693. 1508. 1460. 1383. 1334. 1270. 1236. 1142. 1115.
1068. 990 cm'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 Example 10 (Exemp. compound No. 53)
OH
H
0 H O H'CONH 2
N
HN N 00 N NH
CH
3 d b 0 (10-1) The compound shown above was synthesized in accordance with the process described in Japanese Patent Application Kokai Hei 5-148293. Described specifically. 1 g ofcapuramycin was dissolved in 175 mL of acetone. To the resulting solution were added 9.2 mL of 2.2-dimethoxypropane and 253 mg of "Amberlyst The resulting mixture was stirred at room temperature. Two days later, the "Amberlyst 15 evaporated and the solvent was distilled off under reduced pressure. The residue was dissolved in 7 mL of chloroform, followed by the addition of 30 mL of hexane. White crystals thus precipitated were collected by filtration, and charged on a silica gel column (40 which was developed with methylene chloride methanol whereby 582.7 mg of the following compound was obtained.
0o S H '0 CONH, Y 0 HN N 0 N NH 0
CH
3 0
OH
1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 9.69 (br s. 1 7.93 J 6.0 Hz. 1 7.74 J 8.2 Hz. 1 7.30 (br s. 1 H).
7.03 1H). 6.34 J 4.4 Hz. 1H). 6.12 (br s. 1H). 5.92 J 6.4 Hz. 1H). 5.73 J 8.2 Hz, 1H), 4.82 J 7.2 Hz. 1H). 4.74 1H). 4.69 1H). 4.60 (m.
1H). 4.53 1H), 4.32 1H). 4.13 J 6.5 Hz. 1H). 4.02 IH). 3.69 IH).
3.50 3H). 3.28 2H). 2.18-1.70 6H). 1.49 3H), 1.45 3H) ppm.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 (10-2) In 3 mL of pyridine were dissolved 100 mg of the compound obtained in 243 mg of palmitic anhydride and 2 mg of DMAP. The resulting solution was stirred at room temperature. One hour later, 1 mL of methanol was added to terminate the reaction. The solvent was then distilled off under reduced pressure.
The residue was dissolved in 100 mL of ethyl acetate. After washing with 100 mL of saturated aqueous sodium bicarbonate. drying was conducted over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure. From the residue, pyridine was removed by azeotropy with toluene, whereby a mixture containing the below-described compound was obtained. The mixture was provided for the subsequent reaction (10-3) without purification.
0 0 H a 0
CONH
2 HN N O O N ,NH S O O
CH
3 b 0 0 (10-3) In 10 mL of methanol was dissolved the whole amount of the mixture obtained in To the resulting solution was added 100 mg of"Amberlyst and the mixture was stirred for 47 hours at room temperature and for 4 hours at After filtration through Celite. the solvent was distilled off under reduced pressure. The residue was charged on a silica gel column (5 which was developed with methylene chloride methanol (95:5 to 93:7). whereby 84.9 mg of the belowdescribed compound was obtained as the desired compound of Example
OH
,OH
0 H OHCONH 2
I
H NH O N NH HN N 0
N
CH
3 0
O
0 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 6 7.94 J 8.2 Hz. 1H). 6.01 J 3.5 Hz. 1H). 5.98 J 4.8 Hz. 1H). 5.72 J 8.2 Hz, 1H). 5.42 J 4.8 Hz. 1H). 5.24 J 5.5 Hz. 1H), 4.68 J 1.8 Hz. 1 4.55 2H). 4.42 J 4.1 Hz. 1H). 4.05 J 4.8 Hz. 1H). 3.98 J 4.7 Hz, 1H). 3.38 3H). 3.25 2H). 2.37 J 7.3 Hz. 2H). 2.01 2H). 1.84 2H). 1.63-1.15 28H). 0.90 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3380. 2925. 1854. 1686. 1509. 1466. 1384. 1334. 1270. 1146. 1112. 1062 cm".
Example 11 (Exemp. compound No. 21)
OH
HN N a o O N NH 0 n (11-1) In 1.5L of acetone was dissolved 8.5 g of A-500359A. To the resulting solution were added 72.7 mL of 2.2-dimethoxypropane and 2 g of "Amberlyst The resulting solution was stirred at room temperature. Three days later, the "Amberlyst 15 was filtered off and the solvent distilled off under reduced pressure. The residue was dissolved in 50 mL of chloroform. followed by the addition of 200 mL of hexane. White crystals thus precipitated were collected by filtration and charged on a silica gel column (400 g) which was developed with methylene chloride methanol whereby 8.83 g of the following compound was obtained.
0 Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 1) 1H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 9.90 (br s, 1H), 7.93 J 6.2 Hz, 1H), 7.75 J 8.1 Hz. 1H), 7.30 (br s. 1H).
6.63 1H). 6.33 J= 4.0 Hz. 1H), 6.14 (br s. 1H). 5.93 J 6.2 Hz. 1H). 5.73 J 8.2 Hz. 1H), 4.83 J 7.1 Hz, 1H), 4.70 2H). 4.61 1H). 4.53 (m.
IH), 4.32 IH), 4.12 J 6.6 Hz. 1H), 4.00 1H). 3.55 1H). 3.50 3H).
2.18-1.20 15H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 2986. 2935. 1692. 1509. 1458. 1432. 1383. 1338. 1269. 1252. 1219. 1167.
1118. 1080. 1064. 1012 cm (11-2) In 2 mL of THF were dissolved 125 mg of the compound obtained in (11-1).
68 mg of 3.3-diphenylpropionic acid. 6 mg of DMAP and 58 mg of WSC. The resulting solution was stirred at room temperature. Two hours later, the solvent was distilled off under reduced pressure. The residue was dissolved in 20 ml of methylene chloride. The resulting solution was washed successively with 20 mL of aqueous sodium bicarbonate and 20 mL of 0.0 IN aqueous hydrochloric acid. and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, whereby a mixture containing the below-described compound was obtained. The resulting mixture was provided for the subsequent reaction (11-3) without purification.
0-+ S
NHCONH
2 HN N O iON N ru r I I- I I Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 (11-3) In 5 mL of methanol was dissolved the whole amount of the mixture obtained in To the resulting solution was added 120 mg of"Amberlyst 15 (H and the resulting mixture was stirred at 80 0 C for 3 hours. After filtration through Celite, the solvent was distilled off under reduced pressure. The residue was charged on a silica gel column (15 g) which was developed with methylene chloride methanol (94:6 to 92:8). whereby 107 mg of the below-described compound was obtained as the desired compound of Example 11.
OH
HN N -oNYNH
CH
3 0 0d O 0 1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. H nuclear magnetic resonance spectrum is as follows: 8 7.77 J 8.1 Hz. 1H). 7.24 8H). 7.14 2H). 6.00 J 4.0 Hz. 1H), 5.90 J 5.4 Hz. 1H). 5.65 J 8.1 Hz. 1H). 5.27 J 5.2 Hz. 1H). 5.20 J 5.4 Hz. 1H). 4.65 J 2.1 Hz. IH). 4.50 3H). 4.38 J 4.0 Hz. 1H). 4.00 J 4.6 Hz. 1H). 3.93 J 4.9 Hz. 1H). 3.58 1H). 3.18 3H). 3.14 J 8.1 Hz.
2H). 2.05-1.75 4H). 1.48 1H). 1.25 1H). 1.22 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3380. 2930. 1690. 1510. 1455. 1431. 1384. 1336. 1267. 1149. 1108. 1081. 1062 cm Example 12 (Exemp. compound No. 22)
OH
H CONH 2 r O H N N N NH OCH 3 O OCH 3 0 0
CH
3 0 '0 Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 The reaction was conducted in a similar manner to that described in Example 11 by using 125 mg of the compound obtained in Example (11-1) and 72 mg of 3- (3,4,5-trimethoxyphenyl)propionic acid. whereby 113.6 mg of the below-described compound was obtained as the desired compound of Example 12.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.92 J= 8.1 Hz, 1H). 6.53 2H). 6.01 J 3.8 Hz. IH). 5.91 J 4.5 Hz.
1H). 5.71 J 8.1 Hz, 1H). 5.45 J =4.8 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.67 J =2.0 Hz. 1H). 4.52 2H). 4.42 J 4.1 Hz. 1H), 4.01 J 4.9 Hz. 1H).
3.97 J 4.9 Hz. 1H). 3.81 6H). 3.71 3H). 3.57 1H). 3.29 3H). 2.87 (t.
J 7.3 Hz. 2H). 2.72 J 7.3 Hz. 2H). 2.05-1.75 4H). 1.48 1H). 1.25 (m.
1H). 1.21 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3388. 2933. 1692, 1591. 1509. 1458. 1424. 1384. 1335. 1268. 1239. 1127 cm'.
Example 13 (Exemp. compound No. 23)
OH
0 "OHCONH, O HN Y N O N NH 0 0 CH30 Ob NO 2
CH
3 The reaction was conducted in a similar manner to that described in Example 11 by using 125 mg of the compound obtained in Example (11-1) and 59 mg of 2-(4nitrophenyl)propionic acid. whereby 121.4 mg of the below-described compound was obtained as the desired compound of Example 13.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 8.22 2H), 7.92 1H). 7.55 J 8.6 Hz, 2H), 5.97 2H). 5.72 1H), 5.43 1H), 5.22 1H), 4.68-4.38 4H), 4.08-3.90 3H), 3.57 1H), 3.33 Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 1.5H), 3.12 1.5H), 2.05-1.75 4H), 1.48 4H), 1.30 1H), 1.22 J= 6.6 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3383, 2931, 1691, 1606, 1521, 1458. 1431, 1384, 1348, 1269, 1237. 1205. 1151.
1108. 1077, 1020 cm-.
Example 14 (Exemp. compound No.
OH
\OH
0 H 'rO CONH 2 H N O N NH 0 0
CH
3 b 0 The reaction was conducted in a similar manner to that described in Example 11 by using 125 mg of the compound obtained in Example 145 mg of pentadecanoic acid, 12 mg of DMAP and 116 mg of WSC, whereby 103.2 mg of the below-described compound was obtained as the desired compound of Example 14.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.8 Hz. 1H). 5.97 J 4.9 Hz. 1H). 5.72 J 8.1 Hz, 5.44 J 4.8 Hz. IH). 5.24 J 5.7 Hz. 1H). 4.68 J 1.9 Hz. 4.55 2H). 4.42 J 4.1 Hz. 1H). 4.06 J 4.7 Hz. 1H). 3.97 J Hz. IH). 3.57 1H). 3.38 3H). 2.37 J 7.4 Hz. 2H). 2.05-1.75 4H).
1.63-1.15 29H). 0.90 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3391.2925.2854. 1686. 1510. 1460. 1430. 1384. 1337. 1270. 1235. 1146. 1109.
1061. 1021.978 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 Example 15 (Exemp. compound No. 46)
OH
0 H r HCONH 2 HN N O N NH 0 CH3d b 0 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (10-1) and 129 pL of heptanoic anhydride, whereby 63.7 mg of the compound shown above was obtained as the desired compound of Example 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.2 Hz. 1H). 6.01 J 3.6 Hz. 1H). 5.97 J 4.9 Hz. 1H). 5.72 J 8.2 Hz. 1H). 5.42 J 4.9 Hz. 1H). 5.24 J 5.5 Hz. 1H). 4.68 J Hz. 1H). 4.55 2H), 4.42 J 4.2 Hz. 1H). 4.04 J 4.8 Hz. 1H). 3.98 J 4.9 Hz. 1H), 3.37 3H). 3.25 2H). 2.37 J 7.3 Hz. 2H). 2.00 2H). 1.83 2H). 1.63-1.25 10H). 0.90 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3382. 2930. 2858. 1687. 1510. 1462. 1384. 1334. 1269. 1236. 1156. 1109. 1062 cm".
Example 16 (Exemp. compound No. 11)
OH
o 0 O H OHCONH OY
CH
3 d b 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example 158 mg of palmitic anhydride and 2 mg of DMAP. whereby 9 3 .4 mg of the compound shown above was obtained as the desired compound of Example 16.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.7 Hz. 1H). 5.98 J 4.9 Hz. I 5.72 J 8.1 Hz, 1H), 5.44 J 4.9 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.68 J 1.7 Hz, 1H). 4.55 2H). 4.41 J =4.2 Hz. 1H). 4.06 J =4.8 Hz. 1H). 3.97 J 4.7 Hz, 1H). 3.58 1H). 3.38 3H). 2.37 J 7.3 Hz. 2H). 2.05-1.75 4H).
1.63-1.20 31H). 0.90 J 6.9 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3390. 2925. 2854. 1744. 1689. 1509. 1459. 1432. 1384. 1337. 1269. 1235. 1147.
1111. 1062. 1021 cm".
Example 17 (Exemp. compound No. 7)
OH
,OH 0 0 H OCONH 2 HN N O N NH
CH
3 b
V
0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (11-1) and 177 pL of decanoic anhydride. whereby 62.2 mg of the compound shown above was obtained as the desired compound of Example 17.
1) 1H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.8 Hz, IH). 5.97 J 4.7 Hz. 1H). 5.72 J 8.1 Hz, 1H), 5.44 J 4.9 Hz. 1H). 5.24 J 5.4 Hz. 1H). 4.68 J 1.7 Hz. 1H), 4.55 2H), 4.41 J 4.1 Hz. 1H), 4.06 J 4.8 Hz. 1H). 3.97 J Hz. 1H). 3.58 IH). 3.38 3H). 2.37 J 7.4 Hz. 2H). 2.05-1.75 4H), 1.63-1.20 19H), 0.90 J 6.8 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 3390, 2927. 2855. 1689, 1510. 1459. 1430. 1384. 1336, 1269, 1151. 1109, 1062. 1022 cm.
Example 18 (Exemp. compound No. 6)
OH
0
COHCONH
2 HN N ONY NH 0 N N 0 0
CH
3 C bs 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (11-1) and 160 pL of pelargonic anhvdride. whereby 59.9 mg of the desired compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6= 7.95 J= 8.1 Hz. 1H). 6.01 (d.J=3.8 Hz. 1H), 5.97 J= 4.7 Hz, 1H). 5.72 J 8.1 Hz. 1H), 5.44 J 4.9 Hz. 1H), 5.24 J 5.6 Hz. 1H). 4.68 J 1.6 Hz. 1H). 4.55 2H). 4.42 J 4.1 Hz. 1H). 4.06 J 4.8 Hz. 1H). 3.97 J 4.9 Hz. 1H). 3.58 1H). 3.38 3H). 2.37 J 7.3 Hz. 2H). 2.05-1.75 4H), 1.63-1.20 17H). 0.90 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 2928. 2856. 1688. 1510. 1459. 1384. 1336. 1269. 1153. 1108. 1061. 1023 cm-'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 Example 19 (Exemp. compound No. 9)
OH
0 ONH 2 0 H OH2 N NH HN N Y 0 O 0 CHd 3 0 b 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (11-1) and 105 mg of myristic anhydride. whereby 81.6 mg of the compound shown above was obtained.
1) lH nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.9 Hz. 1H). 5.97 J 4.8 Hz. 1H). 5.72 J 8.1 Hz. 1H). 5.44 J 4.9 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.68 J 1.8 Hz. IH). 4.55 2H), 4.42 J 4.1 Hz. 1H). 4.06 J 4.8 Hz. 1 3.97 J 4.9 Hz. 1H). 3.58 1H). 3.38 3H). 2.37 J 7.3 Hz. 2H). 2.05-1.75 4H).
1.63-1.20 27H). 0.90 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 2925. 2854. 1689. 1509. 1459. 1384. 1337. 1269. 1148. 1110. 1062. 1022 cm Example 20 (Exemp. compound No. 8)
OH
\OH 0 H0 H CON H, HN N oT NKNK'f NH N 0 CH 3 O' t 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (11-1) and 91.8 mg of lauric anhydride, whereby 69.7 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 6 7.95 J 8.2 Hz, 1H). 6.01 J 3.9 Hz, 1H). 5.97 J 4.7 Hz, 1H). 5.72 J 8.2 Hz, 1H), 5.44 J 4.9 Hz, 1H). 5.24 J 5.7 Hz, 1H), 4.69 J 1.6 Hz, 1H), 4.55 2H). 4.42 J 4.1 Hz, 1H), 4.07 J 4.8 Hz. 1H), 3.97 J 4.7 Hz, 1H), 3.58 1H), 3.38 3H). 2.37 J 7.3 Hz, 2H), 2.05-1.75 4H).
1.63-1.20 23H), 0.90 J 7.0 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 2926, 2855. 1689. 1509. 1459. 1384. 1336. 1269. 1149. 1110. 1062. 1022 cm'.
Example 21 (Exemp. compound No. 16)
OH
o OH N NN H 0 0
CH
3 d 0 The reaction was conducted in a similar manner to that described in Example 11 by using 100 mg of the compound obtained in Example (11-1) and 92.2 ml ofoleic acid. whereby 70.9 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.2 Hz. 1H). 6.01 J 3.9 Hz. 1H). 5.97 J 4.8 Hz. 1H). 5.72 J 8.2 Hz, 1H). 5.44 J 4.9 Hz. 1H). 5.34 J 4.8 Hz. 2H). 5.24 J 5.7 Hz. 1H). 4.68 J 1.9 Hz. 1H). 4.55 2H). 4.42 J 4.1 Hz. 1H). 4.07 J 4.8 Hz. 1H). 3.97 J 4.7 Hz. 1H). 3.58 1H). 3.38 3H). 2.37 J 7.4 Hz.
2H). 2.05-1.75 8H). 1.60 2H). 1.49 1H). 1.33 21H). 1.22 J 6.7 Hz. 3H). 0.89 J 7.0 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3391. 2926, 2855, 1688. 1509. 1459. 1431, 1384, 1336, 1269, 1145, 1109. 1061, 1022 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Example 22 (Exemp. compound No. 18)
OH
0 H HCONH 0 HN N N NH CH3O\
O
0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (11-1) and 259 mg of linolenic acid anhydride. whereby 65 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.0 Hz. 1H). 6.01 J 3.8 Hz. 1H). 5.97 J 4.8 Hz. 1H). 5.72 J 8.0 Hz, 1H), 5.45 J 4.9 Hz. 1 5.34 6H). 5.24 J 5.7 Hz. 1 H).
4.68 J 1.9 Hz, 1H). 4.55 2H). 4.41 J 4.2 Hz. 1H). 4.07 J 4.8 Hz.
1H). 3.97 J 4.8 Hz. IH). 3.58 1 3.38 3H). 2.81 J 5.9 Hz. 4H). 2.38 J= 7.3 Hz. 2H). 2.10-1.75 8H). 1.60 2H). 1.49 1H). 1.32 9H). 1.22 J 6.7 Hz. 3H). 0.97 J 7.5 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 3011. 2928. 2855. 1688. 1509. 1459. 1430. 1385. 1337. 1269. 1144. 1108.
1061. 1022 cm-.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-21 8:Examples)/1 8.12.00 Example 23 (Exemp. compound No. 17)
OH
0 OHCONH 2
O
HN H N N NH H N Med b The reaction was conducted in a similar manner to that described in Example by using 150 mg of the compound obtained in Example (11-1) and 326 mg of linoleic anhydride, whereby 80.5 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J= 8.1 Hz. 1H). 6.01 J= 3.9 Hz. 1H). 5.97 J= 4.8 Hz. 1H). 5.72 J 8.1 Hz. IH). 5.45 J 4.9 Hz. 1H). 5.35 4H). 5.24 J 5.7 Hz. 1H).
4.68 J 1.9 Hz. 1H). 4.55 2H), 4.41 J 4.2 Hz. 1H). 4.07 J 4.8 Hz.
1H). 3.97 J 5.0 Hz. 1H). 3.58 1H). 3.38 3H). 2.77 J 6.3 Hz. 2H). 2.38 J 7.3 Hz. 2H). 2.10-1.75 8H). 1.60 2H). 1.49 1H). 1.32 1.22 J 6.7 Hz. 3H). 0.97 J 6.9 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3388. 3009. 2928. 2856. 1687. 1510. 1459. 1430. 1384. 1337. 1270. 1144. 1108.
1061. 1021 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Example 24 (Exemp. compound No.
OH
0 OH CONH 2
O
HN NH 0 N NH MeO' b 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (10-1) and 125.5 mg of lauric anhydride. whereby 78.3 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.9 Hz. 1H). 5.97 J 4.8 Hz. 1H). 5.72 J 8.1 Hz. 1H). 5.42 J 4.9 Hz. 1H). 5.24 J 5.7 Hz. 1H). 4.68 J 1.6 Hz. 1H). 4.55 2H). 4.42 J 4.1 Hz. 1H). 4.04 J 4.8 Hz. 1 3.98 J 4.8 Hz. 1H). 3.37 3H). 3.25 2H). 2.37 J 7.3 Hz. 2H), 2.00 2H). 1.84 2H). 1.64-1.25 20H). 0.90 J 6.8 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3381. 2926. 2855. 1689. 1509. 1462. 1436. 1383. 1333. 1269. 1149. 1111. 1063 cm Example 25 (Exemp. compound No. 49)
OH
o 'OHCONH 2 r HN N 0 O 0 N NH MeO O 0 The reaction was conducted in a similar manner to that described in Example by using 150 mg of the compound obtained in Example (10-1) and 181 pl of decanoic anhydride, whereby 124.3 mg of the compound shown above was obtained.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.1 Hz, 1H), 6.01 J 3.9 Hz, 1H), 5.97 J 4.9 Hz, 1H). 5.72 J 8.1 Hz, 1H), 5.42 J 4.8 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.68 J 1.7 Hz, 1H), 4.55 2H), 4.42 J 4.2 Hz, 1H), 4.04 J 4.8 Hz. 1H). 3.98 J 4.8 Hz. 1H), 3.37 3H), 3.25 2H), 2.37 J 7.3 Hz. 2H). 2.00 2H). 1.84 2H), 1.64-1.25 16H). 0.90 J 6.8 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3378. 2927. 2856. 1689. 1509. 1462. 1436. 1383. 1333. 1270. 1151. 1111. 1063 cm'.
Example 26 (Exemp. compound No. 51)
OH
0 OH O H CONH 2 r HN N 0 O N NH 0 0 Med b 0 The reaction was conducted in a similar manner to that described in Example by using 100 mg of the compound obtained in Example (10-1) and 181 mg of myristic anhydride, whereby 67.5 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6= 7.94 J 8.1 Hz. 1H). 6.01 J 3.9 Hz. 1H). 5.97 J 4.8 Hz. 1H). 5.72 J 8.1 Hz. 1H), 5.42 J 5.0 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.68 J 1.6 Hz. 1H), 4.55 2H), 4.42 J 4.1 Hz. 1H). 4.04 J 4.8 Hz. 1H). 3.98 J 4.9 Hz. 1H), 3.37 3H), 3.25 2H). 2.37 J 7.3 Hz. 2H). 2.00 2H). 1.84 2H). 1.64-1.25 24H). 0.90 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3378. 2926, 2855, 1689, 1509, 1464. 1435, 1383, 1333, 1269, 1147, 1111. 1063 cm-.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 Example 27 (Exemp. compound No. 48)
OH
0 H \OHCONH 2
-Y
H N O N N H b" 0 O The reaction was conducted in a similar manner to that described in Example by using 150 mg of the compound obtained in Example (10-1) and 163 pl of pelargonic acid anhydride. whereby 93.5 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.1 Hz, 1H). 6.01 J 3.8 Hz. 1H). 5.97 J 5.0 Hz. 1H), 5.72 J 8.1 Hz, 1H), 5.42 J 4.8 Hz. 1H). 5.24 J 5.4 Hz, 1H). 4.68 J 1.8 Hz. 1H), 4.55 2H), 4.42 J 4.2 Hz, IH). 4.04 J 4.8 Hz. 1H). 3.98 J 4.9 Hz. 1H), 3.37 3H). 3.25 2H). 2.37 J 7.3 Hz. 2H). 2.00 2H). 1.84 2H). 1.64-1.25 14H). 0.90 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3376. 2927. 2856. 1690. 1509. 1461. 1436. 1379. 1334. 1264. 1150. 1108. 1064 cm'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Example 28 (Exemp. compound No. 282) 0 0 0 H CONH 2 H N N NH HN N O
CH
3 0 bOH The reaction was conducted in a similar manner to that described in Example 9 by using 243 mg of the compound obtained in Example and 130 pl of pelargonic acid anhydride. whereby 145.5 mg of the compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.72 J 8.1 Hz. 1H). 5.99 J 2.5 Hz. 1H). 5.88 J 8.1 Hz. 1H). 5.81 (d.
J 4.5 Hz. 1H). 5.72 1H). 5.64 1H). 5.45 J 3.3 Hz. 1H). 4.68 J 2.2 Hz, 1H). 4.58 (dd, J 1.0 and 10.9 Hz. I 4.46 (dd. J 2.2 and 5.2 Hz. 1 4.18 (t, J 4.8 Hz, 1H). 3.65 J 5.2 Hz. 1H). 3.34 3H). 3.25 2H). 2.37 4H).
2.03 2H). 1.85 2H). 1.62 5H). 1.32 21H). 0.90 6H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3369. 2927. 2856. 1749. 1693. 1508. 1461. 1380. 1335. 1270. 1258. 1143. 1115. 1067 cm Example 29 (Exemp. compound No. 52)
OH
0 H O CONH 2 O H NCH 3 N 0 0 O The reaction was conducted in a similar manner to that described in Example 11 by using 153.7 mg of the compound obtained in Example (10-1) and 122.2 mg of pentadecanoic acid, whereby 102.8 mg of the compound shown above was obtained.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.1 Hz. 1H). 6.01 J 3.7 Hz. 1H), 5.97 J 5.0 Hz. 1H). 5.72 J 8.1 Hz. 1H), 5.42 J 4.9 Hz, 1H), 5.24 J 5.6 Hz. 1H). 4.68 J Hz, 1H). 4.55 2H), 4.42 J 4.1 Hz, 1H). 4.04 J =4.8 Hz. 1H). 3.98 J 4.8 Hz. 1H), 3.37 3H), 3.25 2H). 2.37 J 7.3 Hz. 2H), 2.00 2H). 1.84 2H). 1.64-1.25 26H). 0.90 J 6.8 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3383. 2925. 2854. 1688. 1509. 1465. 1436. 1384. 1334. 1270. 1147. 1112. 1063 cm".
Example 30 (Exemp. compound No. 283) O 0 HN N Y O 0 N YNH 0 0
CH
3 0 bH The reaction was conducted in a similar manner to that described in Example 9 by using decanoic acid anhydride instead of heptanoic acid anhydride. whereby 40.6 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: S= 7.72 J 8.1 Hz. lH). 5.99 1H). 5.87 J 8.1 Hz. 1H). 5.81 4.4 Hz. 1H). 5.72 1H). 5.64 1H). 5.45 J 3.1 Hz. 1H). 4.68 J 2.2 Hz.
1H). 4.57 1H), 4.46 (dd. J 2.1 and 5.4 Hz. 1H). 4.18 J 5.0 Hz. 1H). 3.65 (t, J 5.0 Hz. 1H), 3.33 3H). 3.25 2H), 2.36 4H). 2.02 2H). 1.85 2H), 1.70-1.25 30H), 0.90 J 6.3 Hz. 6H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 3375. 2926. 2854. 1747. 1691. 1507. 1463. 1380. 1334. 1267, 1247. 1142. 1115. 1066
-I
cm.
Example 31 (Exemp. compound No.
OH
SOH
H CONH 2 j N N
NH
O
0o 0
CH
3 6 b 0 The reaction was conducted in a similar manner to that described in Example by using 187 mg of the compound obtained in Example (11-1) and 267 pl of octanoic acid anhydride. whereby 115 mg of the desired compound shown above was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 8 7.95 J 8.1 Hz. 1H). 6.01 J 3.9 Hz. 1H). 5.97 J 4.9 Hz. 5.72 J 8.1 Hz. 1 5.44 J 4.9 Hz. I 5.23 J 5.5 Hz. 1 4.68 J Hz. IH). 4.56 1H). 4.52 1H). 4.42 J 4.1 Hz. 1H). 4.06 J 4.7 Hz. 1H), 3.97 J 5.1 Hz. lH). 3.57 1H). 3.38 3H). 2.37 J 7.3 Hz. 2H). 2.05-1.75 4H). 1.60 2H). 1.48 1H). 1.32 9H). 1.21 J 6.6 Hz. 3H). 0.90 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3399. 2930. 2857. 1686. 1511. 1459. 1430. 1385. 1335. 1268. 1231. 1152. 1107.
1061. 1022 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 Example 32 (Exemp. compound No. 540)
OH
SOH
O H O 0 H CONH fr O I HN- NO O O N NH
CH
3 0 b 0 In 3 mL of pyridine were dissolved 125 mg of the compound obtained in Example 170 pl of nonyl chloroformate. 147 mg of dimethylaminopyridine and 3 mg of 4-pyridylpyridine. The resulting solution was stirred at room temperature. Three hours later, the solvent was distilled off under reduced pressure.
The residue was then dissolved in 60 mL of ethyl acetate. After washing with 60 mL of each of saturated aqueous NaHCO 3 and saturated saline, drying was conducted over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure and the residue was dissolved in 4 mL of methanol. To the resulting solution was added 200 mg of"Amberlyst 15". followed by heating under reflux. Three hours later, the insoluble matter was filtered off and the solvent was distilled off under reduced pressure. The residue was subjected to a silica gel column (8 g) and eluted with 5% methanol methylene chloride. whereby 108 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.2 Hz. IH). 6.01 J 4.0 Hz. 1H). 5.98 J 4.6 Hz. I 5.71 J 8.2 Hz, 1H). 5.32 J 4.8 Hz. 1H). 5.23 J 5.7 Hz. 1H). 4.68 J Hz. 1H). 4.56 1H). 4.52 1H). 4.41 J 4.2 Hz. 1H). 4.13 3H). 3.97 J 5.0 Hz, 1H). 3.57 1H). 3.40 3H). 2.05-1.75 4H). 1.65 2H). 1.48 (m.
1H). 1.32 13H). 1.22 J 6.6 Hz. 3H). 0.90 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3385. 2929, 2855. 1753. 1691. 1510. 1458. 1431. 1393, 1259. 1144. 1101. 1076, 1021 cmcm Doc: FP9907s4.doc P81485/FP-9907 PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)iI 8.12.00 Example 33 (Exemp. compound No. 539)
OH
SOH
00 0 H
CONH
2 0 HN NO N
NH
0
CH
3 d
O
The reaction was conducted in a similar manner to that described in Example 32 except for the use of 157 pl of octyl chloroformate instead of nonyl chloroformate.
whereby 91 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.94 J 8.1 Hz, 1H). 6.01 J 3.9 Hz. 1H). 5.98 J 4.4 Hz. 1H). 5.71 J 8.1 Hz, 1H), 5.32 J 4.6 Hz. 1H). 5.24 J 5.6 Hz. 1H). 4.69 J Hz. 1H), 4.56 1H), 4.52 1H). 4.41 J 4.0 Hz. 1H). 4.13 3H). 3.97 J 5.0 Hz. 1H). 3.57 1H). 3.40 3H). 2.05-1.75 4H). 1.65 2H). 1.48 (m.
1H), 1.32 11H), 1.22 J 6.6 Hz. 3H). 0.90 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3387. 2929. 2856. 1752. 1689. 1510. 1458. 1431. 1392. 1335. 1260. 1143. 1101.
1073. 1021 cm-.
Example 34 (Exemp. compound No. 594)
OH
,OH
,n
CH
3 6 Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218.Examples)/l 8.12.00 (34-1) 0 I CONH 2 0 HN N 0 0 N N O CH3CO bH In 50 mL of dimethylformamide (DMF) were dissolved 4.57 g of the compound obtained in Example (11-1) and 2.2 mL of 1.8-diazabicyclo[5.4.0]-7undecene (DBU). To the resulting solution was added a solution obtained by dissolving 2.45 g of4-methoxybenzyl chloromethyl ether in 50 mL of DMF. The resulting mixture was stirred at room temperature. After 2.5 hours. the solvent was distilled off under reduced pressure. The residue was dissolved in 300 mL of methylene chloride. The resulting solution was washed successively with 300mL each of 0.01N aqueous hydrochloric acid. saturated aqueous sodium bicarbonate and saturated saline, and then dried over anhvdrous magnesium sulfate. The solvent was distilled off under reduced pressure and then charged on a silica gel column (200 g), which was developed with 3°%o methanol in methylene chloride, whereby 4.80 g of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follow\s: 6 7.85 IH). 7.69 J 8.2 Hz. 1H). 7.32 2H). 7.15 2H). 6.85 J 8.7 Hz. 2H). 6.37 J 4.3 Hz. I 6.06 J 6.2 Hz. 1 5.82 1 5.75 J 8.2 Hz. IH), 5.70 1H). 5.44 2H). 4.73 3H). 4.61 2H). 4.57 IH). 4.45 1H). 4.25 1H). 4.03 2H). 3.79 3H). 3.56 3H). 3.53 1H). 3.28 (d.
J 7.8 Hz. 1H). 2.35 21H 2.15 1H). 2.02-1.75 4H). 1.49 3H). 1.42 (s.
3H), 1.30 2H), 1.23 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3387, 3105, 2984, 2935. 1669. 1612. 1514, 1457. 1383, 1361, 1300. 1248. 1219.
1169. 1114, 1079, 1064, 1012 cm-.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 (34-2) 0 H0 CONH 2 0 0 HN N N N O 0
CH
3 d b In 5 mL of DMF was dissolved 773 mg of the compound obtained in Example The resulting solution was stirred at 0°C under a nitrogen gas stream. To the reaction mixture was added 60 mg of NaH (about Two minutes later. 2.13 mL of 1 -iododecane was added. Five minutes later, the temperature was allowed to rise back to room temperature. at which stirring was conducted for further 25 minutes.
The reaction mixture was then distilled under reduced pressure to remove the solvent.
The residue was dissolved in 250 mL of methylene chloride. The resulting solution was washed successively with 300 mL each of 0.01N aqueous hydrochloric acid.
saturated aqueous sodium bicarbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure and the residue charged on a silica gel column (200 g) which was developed with 2% methanol in methylene chloride. whereby 395 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 1H nuclear magnetic resonance spectrum is as follows: 6 7.89 J 8.1 Hz. IH). 7.75 J 5.9 Hz. IH). 7.31 (d.J 8.8 Hz. 2H). 7.13 (br s. 1H). 6.86 J 8.8 Hz. 2H). 6.37 IH). 5.95 1H). 5.75 (br s. 1H). 5.70 J 8.1 Hz. 1H). 5.57 1H). 5.45 2H). 4.78 J 8.1 Hz. IH). 4.74 2H).
4.63 2H), 4.55 1H). 4.46 1H). 4.05 2H). 3.95 1H). 3.79 3H). 3.62 1H). 3.51 1H). 3.43 3H). 4.09 1H). 1.98 IH). 1.86 1H). 1.77 1H). 1.49 3H). 1.44 3H). 1.40-1.20 18H). 1.19 J 6.6 Hz. 3H). 0.88 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218 Examples)/I 8.12.00 3386.3102. 2928. 2855. 1713. 1670. 1613. 1587. 1514. 1456. 1382. 1359. 1338.
1300. 1271. 1248. 1220. 1167. 1112. 1066, 1013 cm 1 (34-3) 00 O H CONH 2 HN N 0 N NH 0 0
CH
3 c b In 5 mL of methylene chloride was dissolved 390 mg of the compound obtained in Example To the resulting solution were added 276 uL of water and 484 mg of 2.3-dichloro-5.6-dicyano-1.4-benzoquinone and the resulting mixture was stirred at room temperature. After 75 minutes, the insoluble matter was filtered off. The filtrate was diluted with 200 mL of methylene chloride, followed by successive washing with 200 mL each of saturated aqueous sodium bicarbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure and the residue was charged on a silica gel column (50 g) which was developed with 5% methanol in methylene chloride.
whereby 278 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 9.30 (brs. 1H). 7.99 J 7.3 Hz. 1H). 7.70 J 8.1 Hz. 1H). 7.19 (br s. 1H), 6.36 J 4.4 Hz. 1H). 5.98 (br s. 1H). 5.85 (brs. 1H). 5.81 J 5.1 Hz. 1H). 5.69 (dd. J 2.2 and 8.1 Hz. 1H). 4.74 2H). 4.60 2H). 4.28 J 4.7 Hz. 1H). 4.12 J 6.2 Hz. 1H). 4.07 J 4.7 Hz. 1H). 3.59 3H). 4.43 3H). 2.10-1.73 (m, 4H). 1.60 2H), 1.48 3H). 1.42 3H). 1.23 19H). 0.88 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3387, 3227, 3098, 2928. 2855, 1692, 1506, 1457, 1431, 1382, 1337, 1296, 1268, 1250. 1235, 1220, 1166, 1121, 1082, 1065. 1013 cm 1 Doc: FP9907s4.doc P81485/FP-9907( PCT )/tsa-ad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 (34-4)
OH
SOH
O H CONH 2 r O HN NO O N NH
CH
3 d b In 15 mL of methanol was dissolved 273 mg of the compound obtained in Example To the resulting solution was added 260 mg of"Amberlyst 15" and the resulting mixture was stirred at 80°C. After 4 hours and 20 minutes, the insoluble matter was filtered off. The filtrate was distilled under reduced pressure. and the residue was charged on a silica gel column (15 g) which was developed with methanol in methylene chloride. whereby 176 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: S= 7.95 J 8.1 Hz. 1H). 6.02 J 3.6 Hz. 1H). 5.92 J 4.5 Hz. 1 5.72 J 8.1 Hz. 1H). 5.23 J 5.3 Hz. 1H). 4.67 1H). 4.59 1H). 4.52 1H).
4.38 J 4.2 Hz. 1H). 4.08 J 4.6 Hz. I 3.98 J 4.7 Hz. 1H). 3.94 J 4.7 Hz. 1H). 3.58 3H). 3.40 3H). 2.05-1.75 4H). 1.52 3H). 1.25 (m.
18H). 0.89 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3391. 3099. 2927.2854. 1686. 1509. 1458. 1431. 1385. 1335. 1269. 1132. 1099.
1063. 1020 cm'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 Example 35 (Exemp. compound No. 590)
OH
OH
0 H CONH 2
O
II H rI HN N 0 N NH 0 0
CH
3
O'
(35-1) 0 CONH 2 0 .O 0 HN O N N O S0 0 o 6
CH
3 d b In a similar manner to that described in Example (34-2) except for the use of 1.48 mL of 1-iodohexane instead of 1-iododecane. 460 mg of the desired compound was obtained.
1) 1H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.91 J 8.3 Hz. 1H). 7.24 J 8.6 Hz. 2H). 6.85 J 8.6 Hz. 2H). 6.18 J 4.1 Hz. 1H). 5.92 J 4.0 Hz. 5.74 J 8.3 Hz. 1H). 5.42 2H).
5.11 J 5.4 Hz. 1H). 4.80 IH). 4.70 1H). 4.55 3H). 4.37 J 5.8 Hz, 1H), 4.08 J 4.3 Hz. 1H). 3.94 J 5.2 Hz. IH). 3.76 3H). 3.60 3H). 3.41 3H). 2.05-1.75 4H). 1.55 3H). 1.43 6H). 1.25 8H). 1.19 J 6.6 Hz, 3H), 0.88 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3381. 3103, 2933. 2871, 2859. 1670. 1613. 1587. 1514. 1455. 1383. 1359. 1300.
1271. 1249. 1220, 1167, 1130. 1112. 1066. 1013 cm Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 183 (35-2) '0 0 H CONH 2 0 HN N N NH CH3d b The reaction was conducted in a similar manner to that described in Example (34-3) using 458 mg of the compound obtained in Example 313 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 9.28 (br s, 1H), 7.99 J 6.6 Hz. 1H). 7.71 J 8.1 Hz. 1H). 7.19 (br s. 1H), 6.36 J 4.4 Hz, 1H), 5.98 (br s. 1H). 5.85 (br s. 1H), 5.81 J 5.1 Hz. 1H). 5.69 (dd. J= 2.2 and 8.1 Hz. 1H). 4.74 2H). 4.60 3H). 4.28 J 4.7 Hz. 1H). 4.12 J 6.9 Hz. 1H), 4.07 J 4.7 Hz. 1H). 3.59 3H). 4.42 3H). 2.10-1.73 (m, 4H), 1.60 2H). 1.48 3H). 1.42 3H). 1.23 11H). 0.87 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3386. 3097. 2933.2872. 2859. 1692. 1507. 1457. 1432. 1383. 1337. 1268. 1235.
1220. 1166. 1129. 1082. 1065. 1012 cmn (35-3)
OH
.OH
0 H
CONH
2 HN NO O 0 N NH O O O In 15 mL of methanol was dissolved 273 mg of the compound obtained in Example To the resulting solution was added 2 6 0 mg of"Amberlyst 15". The resulting mixture was stirred at 80 0 C. After 4 hours and 20 minutes, the insoluble Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 matter was filtered off. The filtrate was distilled under reduced pressure. The residue was subjected to a silica gel column (15 g) and then eluted with 5% methanol in methylene chloride, whereby 176 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J= 3.9 Hz. 1H). 5.92 J 4.5 Hz. 1H). 5.72 J 8.1 Hz. 1H). 5.23 J 5.6 Hz. 1H), 4.66 J 2.0 Hz. IH). 4.59 1H).
4.50 1H). 4.38 J 3.9 Hz. 1H). 4.08 J 4.7 Hz. 1H). 3.99 J 4.9 Hz.
1H). 3.93 J 4.7 Hz. 1H). 3.58 3H). 3.40 3H). 2.05-1.75 4H). 1.52 (m.
3H). 1.25 7H). 1.22 J 6.6 Hz. 3H). 0.89 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3387. 3098. 2931.2859. 1687. 1509. 1458. 1431. 1385. 1335. 1268. 1131. 1098.
1063. 1020 cm'.
Example 36 (Exemp. compound No. 891) OH OH H HN N 0 N NH O O
CH
3 0 OH (36-1) BzO O :Bz 0 CONH 2
O
HN N Ora 0 N NH bBz In pyridine was dissolved 300 mg of Compound A-500359A. To the resulting solution were added 696 mg of benzoic anhydride and 6.4 mg of dimethylaminopyridine. The resulting mixture was stirred at room temperature. Four hours later, the solvent was distilled off under reduced pressure and the residue dissolved in 200 mL of ethyl acetate. The resulting solution was washed successively Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 with 200 mL each of saturated aqueous sodium bicarbonate and saturated and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure and the residue was charged on a silica gel column (50 which was developed with 3% methanol in methylene chloride, whereby 423 mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 9.40 (br s. 1H), 8.06 4H). 7.92 4H). 7.55 5H), 7.40 5H). 7.15 (br s. 1H). 6.45 (br s. 1H). 6.32 J 3.7 Hz. 1H). 6.13 1H), 6.09 (br s. IH). 5.96 (d.
J =3.7 Hz. 1H). 5.83 2H). 5.62 2H). 4.69 1H), 4.61 1H). 4.56 (m.
1H). 4.36 J 5.9 Hz. 1H). 3.54 1H). 3.34 3H), 2.12 1H). 2.00-1.50 (m.
4H). 1.32 1H). 1.24 J 6.6 Hz. 3H) ppm.
(36-2) BzO OBz c rT o Y° 0 0
CH
3 O ObBz In 6.3 mL of methylene chloride was dissolved 418 mg of the compound obtained in Example To the resulting solution was added 5 mL of water, followed by stirring at room temperature. To the reaction mixture. 4.74 g of nitrosvlsulfuric acid was gradually added over 30 minutes. After stirring for a further minutes, the resulting mixture was diluted with 30 mL of methylene chloride. The organic layer separated was washed with 10 mL each of water and saturated saline and the solvent was then distilled off under reduced pressure. The residue was dissolved in 10 mL of methylene chloride. To the resulting solution was added an ether solution of diazomethane prepared by mixing 144 mg of N-methyl-Nnitrosourea. 90 mg of potassium hydroxide. 2.8 mL of ether and 2.8 mL of water and the resulting mixture was stirred at room temperature. One hour later, the solvent was distilled off under reduced pressure. The residue was charged on a silica gel column g) which was developed with 1.5% methanol in methylene chloride, whereby 99 mg of the desired compound was obtained.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 8.28 1H), 8.06 J 7.3 Hz. 2H). 7.99 J 7.3 Hz. 2H). 7.95 3H). 7.60- 7.32 I 6.33 1H). 6.20 J 3.6 Hz. 1H). 6.06 J 4.4 Hz. 1H). 5.94 (d.
J 5.9 Hz. 1H), 5.88 J 4.0 Hz. 1H). 5.70 J 3.7 Hz. 1H). 5.54 2H). 4.79 1H). 4.63 1H). 4.17 J 5.5 Hz. 1H). 3.83 3H). 3.80 IH). 3.72 (m.
1H). 3.35 IH). 3.30 3H). 2.19 1H). 2.02-1.75 3H). 1.52 1H). 1.32 I 1.24 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3388. 3093.3069.2933. 2855. 1729. 1697. 1658. 1602. 1584. 1551. 1509. 1452.
1383. 1336. 1315. 1270. 1177. 1115. 1070. 1026 cm-' (36-3) OH OH
H
O 0
O
H r HN N 0 0 N NH HN O O 0 0
CH
3 0C OH In 2 mL of a 40% methvlamine methanol solution was dissolved 98 mg of the compound obtained in Example The resulting solution was hermetically sealed and then stirred. Forty-five minutes later. the solvent was distilled off under reduced pressure. The residue was subjected to reverse-phase preparative HPLC (Inertsil Prep-ODS), followed by elution with 16% acetonitrile water, whereby mg of the desired compound was obtained.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.86 J 8.0 Hz. 1H). 5.98 1H). 5.83 1H). 5.74 (dd. J 2.9 and 8.1 Hz, 1H). 5.24 J 4.9 Hz. 1H). 4.73 (dd. J 2.1 and 10.9 Hz. 1H). 4.50 2H). 4.38 J 4.0 Hz. 1H). 4.25 1H). 4.04 2H). 3.75 1H), 3.39 J 2.8 Hz, 3H).
2.74 J 2.4 Hz, 3H), 1.65 1H). 1.25 2H), 1.00 3H), 0.92 1H), 0.75 2H) ppm.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 Example 37 (Exemp. compound No. 991) OH OH
H
HN O 0 O HN N 0 0 N NH
CH
3 O'
OH
The reaction was conducted in a similar manner to that described in Example (36-3) by using 120 mg of the compound obtained in Example 0.4 mL of npropylamine and 2 mL of methanol. whereby 16 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.91 J 8.1 Hz, 1H). 6.02 J= 4.2 Hz. 1H). 5.89 J 5.5 Hz. IH). 5.72 J 8.1 Hz. 1H), 5.16 J 6.4 Hz. 1H). 4.67 J 2.0 Hz. 1H). 4.55 2H).
4.37 J 4.3 Hz, 1H), 4.33 J 5.2 Hz. 1H). 3.92 2H). 3.60 1H). 3.45 (s.
3H). 3.25 2H). 2.05-1.75 4H). 1.53 3H). 1.25 1H), 1.22 J 6.6 Hz.
3H). 0.91 J 7.5 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3369. 3098, 2964. 2934. 878. 1683. 1515. 1459. 1432. 1385. 1335. 1269. 1140.
1080. 1062. 1022. 981 cm'.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 Example 38 (Exemp. compound No. 1091) OH OH oH 0 H
O
HN N N NH S 0 O.O CH3d bH The reaction was conducted in a similar manner to that described in Example (36-3) using 270 mg of the compound obtained in Example 1.92 g of dodecylamine and 6.9 mL of methanol. whereby 15 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. lH nuclear magnetic resonance spectrum is as follows: S 7.92 J 8.1 Hz. 1H). 6.02 J 4.4 Hz. IH). 5.91 J 5.9 Hz. 1H). 5.73 J 8.1 Hz, 1H). 5.15 J 5.9 Hz. 1H). 4.67 J 2.2 Hz. IH). 4.55 2H).
4.36 J 4.4 Hz. 1H). 4.32 J 5.5 Hz. 1H). 3.92 2H). 3.60 1H). 3.47 (s.
3H). 3.35 1H). 3.20 1H). 2.05-1.75 4H). 1.50 3H). 1.28 19H).
1.22 J 6.6 Hz. 3H). 0.89 J 6.6 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3351. 3098. 2926. 2854. 1685. 1512. 1459. 1432. 1385. 1335. 1264. 1139. 1090.
1063. 1022, 993 cm'.
Example 39 (Exemp. compound No. 548)
OH
r L u b 0 Doc: FP9907s4.doc P81485/FP-9907( PCT/tsa-gad-sh/English translation of spec (pages 135-218: Examples)/ 8.12.00 (39-1) 00 o 0 r"H CONH 2 S H O O N NH HN N o 0 0
CH
3 0 0 In 4 mL of pyridine was dissolved 125 mg of the compound obtained in Example Under a nitrogen gas stream. 147 mg of dimethylaminopyridine and 3.9 mg of 4-pyrrolidinopyridine were added to the solution. After cooling to O'C.
209.1 mg of 2.2-dimethyldodecanoyl chloride Roth. et al. Journal of Medicinal Chemistry. 35. 1609-1617 (1992)) was added. The resulting mixture was stirred at room temperature for 28 hours. After cooling to 0 0 C. 2 mL of methanol was added to the reaction mixture. The resulting mixture was stirred for 10 minutes. followed by concentration under reduced pressure. To the residue were added 20 mL of 0.02N hydrochloric acid and 20 mL of methylene chloride to separate it into layers. The organic layer thus obtained was washed three times with saturated saline, dried over anhvdrous sodium sulfate and concentrated under reduced pressure. whereby 307 mg of a crude product was obtained. The product was purified by Lobar's silica gel column (eluted first with a 3:7 mixture of hexane and ethyl acetate. followed by ethyl acetate). whereby 132 mg of the desired compound was obtained as a white powder.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.90 J 8.1 Hz. 1H). 6.16 J 3.7 Hz. 1H). 6.03 J 5.4 Hz. 1H). 5.72 J 8.1 Hz. 1H). 5.32 J 5.2 Hz, IH). 5.14 J =5.3 Hz. IH). 4.90 IH).
4.75 J 2.1 Hz. IH). 4.59-4.55 2H). 4.38 J 5.8 Hz. i 4.05 J 4.4 Hz, lH), 3.64-3.55 1H). 3.40 3H). 2.01-1.77 4H). 1.59-1.47 3H). 1.45 6H), 1.34-1.10 26H). 0.89 J 6.7 Hz, 3H) ppm.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 8.12.00 (39-2)
OH
SOH
0 H CONH2 O HN N N H 0 0
CH
3 O b
O
To 125 mg of the compound obtained in Example (39-1) was added 50 mL of a 5% trifluoroacetic acid methylene chloride solution. and the resulting mixture was stirred at room temperature for 5 hours. Concentration of the reaction mixture and azeotropy with toluene yielded 147 mg of a crude product. The resulting product was purified by thin-layer chromatography (elution with a 8% methanol in methylene chloride mixture). whereby 64.8 mg of the desired compound was obtained as a white powder.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.02 J 3.9Hz. 1H). 5.98 J 4.8 Hz. 1H). 5.71 (d.
J 8.1 Hz. 1H). 5.39 J 4.8 Hz. 1H). 5.24 J 5.4 Hz. 1H). 4.69 J 2.1 Hz.
1H). 4.57-4.56 1H). 4.54-4.50 1H). 4.42 J 4.1 Hz. 1H). 4.06 J 4.8 Hz. 1H 3.98 J 4.9 Hz. 1H). 3.61-3.53 1H). 3.37 3H). 2.04-1.76 4H), 1.56-1.43 2H). 1.33-1.16 27H). 0.89 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3390. 2927. 2854. 1688. 1510. 1459. 1387. 1336. 1269. 1144. 1108. 1062 cm' Doc: FP9907s4.doc P8 485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/1 8.12.00 Example 40 (Exemp. compound No. 574)
OH
.OH
0 H CONH 2
O
O O HN N 0K0N NH 0 0
CH
3 d b (40-1) ,o 0 0 H CONH 2 HN N 0 N NH 0 0H 0 CH3dC b In a similar manner to that described in Example (39-1) except for the use of 122 mg of the compound obtained in Example (10-1) instead of the compound obtained in Example the reaction was conducted. whereby 126.9 mg of the desired compound was obtained as a white powder.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: S 7.90 J 8.1 Hz. 1H). 6.16 J =3.7 Hz. 1H). 6.03 J 5.7 Hz. 1H). 5.72 J 8.1 Hz. 1H), 5.30 (t.J 5.3 Hz. 1H). 5.15 J =5.4 Hz. IH). 4.90 1H).
4.75 J 2.1 Hz. 1H). 4.59-4.57 2H). 4.39 J 5.9 Hz. 1H). 4.03 J 4.4 Hz. 1H), 3.39 3H). 3.31-3.28 2H). 2.02 J 11 Hz. 2H). 1.87-1.77 2H).
1.60-1.49 2H), 1.44 6H). 1.40-1.20 18H), 1.17 6H). 0.89 J 6.9 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3377, 2929, 2856, 1695, 1507, 1459, 1382, 1334, 1269, 1140, 1116, 1064 cm-.
(40-2) Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218.Examples)/1 8.12.00
CH
3 d b In a similar manner to that described in Example (39-2) except for the use of 95.3 mg of the compound obtained in Example (40-1) instead of the compound obtained in Example whereby 72.4 mg of the desired compound was obtained as a white powder.
1) H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.2 Hz. 1H). 6.02 J 3.8 Hz. 1H). 5.98 J 4.8 Hz. 1H). 5.72 J 8.2 Hz, 1H). 5.37 J 5.0 Hz. 1H). 5.24 J 5.4 Hz. 1H). 4.68 J 2.1 Hz. 1H). 4.57-4.52 2H). 4.42 J 4.1 Hz. 1H). 4.04 J 4.9 Hz. 1H). 3.98 J 4.8 Hz. 1H), 3.37 3H). 3.27-3.22 2H). 2.04-1.89 2H). 1.86-1.77 2H), 1.58-1.46 2H), 1.43-1.19 18H). 1.16 J 6.2 Hz. 6H), 0.89 J 6.9 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) tablet method exhibits absorption maxima as follows: 3369. 2927. 2854. 1689. 1509. 1463. 1389. 1332. 1269. 1143. 1110. 1062 cm'.
Example 41 (Exemp. compound No. 545)
OH
OH
0 O
CH
3 0 b In a similar manner to that described in Example 25 except for the use of 2methyldodecanoyl chloride [synthesized by chlorinating 2-methyldodecanoic acid Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/ 18.12.00 which was synthesized by the process described in Organic Synthesis. 4. 616. by the method as described in B.D. Roth. et al.. Journal of Medicinal Chemistry. 35. 1609- 1617 (1992)] instead of 2.2-dimethyldodecanoyl chloride. 82.5 mg of the desired compound was obtained as a white powder.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.96 J 8.1 Hz, 1H). 6.01 J 4.0 Hz. 1H). 5.98 (dd. J 4.5 and 3.4 Hz.
IH). 5.71 J= 8.1 Hz. 1H). 5.46-5.43 1H). 5.24 J 5.5 Hz. IH). 4.68 J= 1.9 Hz. 1H). 4.57 (dd J 4.8 and 1.7 Hz. 1H). 4.52 (dd. J 11 and 1.5 Hz. 1H). 4.42 J 4.1 Hz. 1H). 4.08-4.05 1H). 3.97 J 5.0 Hz. IH). 3.61-3.54 1H).
3.38 3H). 2.53-2.48 2.04-1.37 6H). 1.28 18H). 1.22 J 6.6 Hz.
3H). 1.15-1.13 3H). 0.89 J 6.8 Hz. 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3389. 2927. 2854. 1689. 1510. 1459. 1384. 1335. 1269. 1145. 1108. 1061 cm Example 42 (Exemp. compound No. 571)
OH
SOH
HN N 0 0 N NH H0 H 3 o o 0 0 CH3-d b In a similar manner to that described in Example 40 except for the use of 2methyldodecanoyl chloride instead of 2.2-dimethyldodecanoyl chloride. 77.5 mg of the desired compound was obtained as a white powder.
1) 'H nuclear magnetic resonance spectrum was measured in deuterated methanol with tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 6 7.95 J 8.1 Hz. 1H). 6.01 J 3.7 Hz, 1H), 5.98 (dd, J 4.5 and 3.6 Hz.
1H), 5.72 J 8.1 Hz, 1H). 5.44-5.40 1H), 5.24 J 5.5 Hz, 1H). 4.68 J 1.8 Hz. 1H), 4.57-4.52 2H). 4.42 J 4.1 Hz, 1H), 4.04 J 4.8 Hz. 1H). 3.98 Doc: FP9907s4.doc P81485/FP-9907( PCT)/tsa-ead-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 J 5.0 Hz. 1H), 3.37 3H). 3.29-3.23 2H). 2.23-2.48 1H), 2.03-1.99 (m.
2H), 1.89-1.76 2H). 1.67-1.32 2H), 1.28 18H), 1.15-1.13 3H), 0.89 J 6.8 Hz, 3H) ppm.
2) Infrared absorption spectrum: The infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits absorption maxima as follows: 3369, 2927, 2854, 1689, 1509. 1461. 1382. 1333. 1269. 1144. 1110. 1062 cm-.
Example 43 Cultivation of Strepiomyces griseus Strain SANK60196 (FERM BP- 5420) Into each of four 2 L Erlenmever flasks (seed flasks), each containing 500 ml of the seed culture medium described below, were inoculated aseptically four loopfuls of Strain SANK60196 followed by shaking in a rotary shaker at 23 0 C and 210 rpm.
and the seed culture was thus conducted for 3 days.
Medium for seed culture: containing the following components in 1000 ml of tap water: Maltose 30 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g Antifoamer "CB442" 50 mg (product of NOF Corporation) After adjustment of pH to 7.4. sterilization was conducted at 121 C for minutes.
Cultivation was conducted as described below. Described specifically, the seed culture was inoculated at (volume/volume: which will hereinafter be abbreviated as into two 30 L jar fermenters. each containing 15 L of a cultivation medium. Six hours later after the initiation of cultivation at 23°C. filtersterilized S-(2-aminoethyl)-L-cysteine hydrochloride was added to give a final concentration of 10 mM. followed by cultivation with aeration and agitation for 6 days.
Medium for cultivation: containing the following components in 1000 ml of tap water: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Maltose 30 g Yeast extract 5 g (product of Difco Laboratories) Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g Antifoamer "CB442" 50 mg (product of NOF Corporation) After adjustment of pH to 7.4. sterilization was conducted at 125°C for minutes.
Example 44 Purification of Compound A-500359E The cultured broth (30 L) obtained in Example 43 was filtered with the aid of "Celite 545" (product of Celite Corporation).
Upon purification as described later, the active fraction was monitored by HPLC using the column and analytical conditions described below.
Column "Senshu Pak ODS-H-2151" 6 x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 0.04°0 aqueous trifluoroacetic acid containing 4% acetonitrile Flow rate: 1.0 ml'min Detection: UV 210 nm Retention time: 21.2 minutes L of the resulting filtrate was charged on a column (6 L) packed with "Diaion HP-20" (product of Mitsubishi Chemical). After washing the column with 12 L ofdeionised water, the non-adsorbed fraction and washing fraction were combined (the combined fraction will hereinafter be called "non-adsorbed-washing fraction").
The adsorbed substance was eluted with 12 L of 10% aqueous acetone. The eluate was concentrated to remove acetone and lyophilized. whereby 39 g of a crude powdery product was obtained.
The resulting crude powdery product was dissolved in 200 mL of deionised water and charged on a column (2 L) packed with "Diaion CHP-20P" (product of Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Mitsubishi Chemical). The column was then washed with 4 L of deionised water and 4 L of 10% aqueous methanol. while the adsorbed substance was eluted with 4 L of aqueous methanol and 4 L of 20% aqueous methanol. A 2 to 4 L portion of the aqueous methanol eluate and the 20% aqueous methanol eluate were combined.
followed by concentration. After removal of methanol by distillation, the residue was lyophilized to give 8.9 g of a powder.
The resulting powder was dissolved in 200 ml ofdeionised water and the resulting solution was charged on a column (1 L) packed with "Toyopearl (product of TOSOH Corporation). followed by development of the column with deionised water. As a result of fractionation of the eluate into portions of 100 ml each. the active substance having a retention time of 21.2 minutes upon the abovedescribed HPLC was eluted in Fraction Nos. 5 to 10. The resulting fractions were concentrated and lyophilized to give 2.7 g of a powder.
The resulting powder was dissolved in 200 ml of deionised water and charged on an HPLC column ("YMC-Pack ODS-1050-20-SR": 100p x 500 mm: product of YMC) equilibrated with 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile. The column was developed at a flow rate of 208 ml/min with 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile. As a result of fractionation of the eluate into portions of 1 L each. the active substance was eluted in Fraction Nos. 6 and 7.
These fractions were combined, followed by concentration to 200 ml by "Evapor" (product of Okawara Seisakujo) and lyophilization. whereby 99 mg of a powder was obtained. The resulting powder was suspended in 5 ml of distilled water and insoluble matter was then filtered off. The filtrate was concentrated to 2 ml by a rotary evaporator, followed by lyophilization. whereby 87 mg of Compound A- 500359E was obtained as a pure product.
The compound A-500359E has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol. insoluble in normal hexane and chloroform 3) Molecular formula: Ci 8 sH 3
N
3 012 4) Molecular weight: 473 (as measured by FAB mass spectrometry) Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 Accurate mass. as measured by high-resolution FAB mass spectrometry is as follows: Found: 474.1349 Calculated: 474.1359 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 251 nm (E 10,000) 7) Optical rotation: optical rotation measured in water exhibits the following value: [aC]D:0 +1150 (c 0.28) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3410. 2955. 1683, 1464. 1441. 1396. 1309, 1267. 1206, 1138. 1115. 1088. 1062. 1023 cm 9) 'H nuclear magnetic resonance spectrum was measured in deuterated dimethyl sulfoxide with tetramethvlsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 3.24 (3H. 3.52 (1H, dd. J=4.5. 6.1Hz), 3.72 (3H. 3.98 (1H. 4.10 (1H. m), 4.25 (1H. 4.29 (1H. d. J=2.0Hz). 4.33 (1H. dd. J=2.0. 6.1Hz). 5.05 (1H. d. J=3.9 Hz). 5.16 (1H. d. J=6.8Hz). 5.45 (1H. d. J=4.2Hz). 5.54 (1H. d. J=5.9Hz). 5.61 (1H. d.
J=3.3Hz). 5.61 (1H. d. J=8.1 Hz). 5.93 (IH. dd. J=1.3. 2.9 Hz). 7.56 (1H. br. 7.69 (IH. br. 7.74 (1H. d. J=8.1 Hz) ppm.
'3C nuclear magnetic resonance spectrum was measured in deuterated dimethvl sulfoxide with tetramethvlsilane as an internal standard substance. 3C-nuclear magnetic resonance spectrum is as follows: 52.0 57.3 61.5 64.9 72.1 75.4 78.2 81.3 89.0 99.2 101.2 114.2 139.2 139.8 150.3 161.8 163.1 170.1 (s)ppm.
11) High performance liquid chromatography (which will hereinafter be abbreviated as "HPLC") analysis: Column: "Senshu Pack ODS-H-2151" 6( x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile Flow rate: 1.0 ml/min Doc: FP9907s4doc P81485/FP-9907(PCT/isa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 Detection: UV 210 nm Retention time: 21 minutes Example 45 Purification of Compounds A-500359F and A-500359H In the purification described below, the active fraction was monitored by HPLC using the following column and analytical conditions.
Column "Senshu Pak ODS-H-2151" 64 x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 mlimin Detection: UV 210 nm Retention time: 8 minutes (Compound A-500359H) 18 minutes (Compound A-500359F) After 42 L of non-adsorbed-washing fraction obtained in Example 44 was adjusted to pH 9 with 6N sodium hydroxide, this fraction was charged on a column L) packed with "Diaion PA316 (product of Mitsubishi Chemical). The column was washed with 27 L of deionised water and the adsorbed substance was then eluted with 27 L of 0.1N hydrochloric acid.
The eluate was adjusted to pH 7 with 6N sodium hydroxide and then charged on an activated charcoal column (2 The column was washed with 8 L of deionised water and the active substance was then eluted with 8 L of 0.5N aqueous ammonia containing 1 0% acetone. Concentration and lyophilization of the resulting eluate yielded 28 g of a powder.
The resulting powder was dissolved in 400 ml of distilled water. After adjustment to pH 3.0. the resulting solution was charged on a column (2 L) which had been adjusted with water and packed with "Diaion CHP-20P" (product of Mitsubishi Chemical). The non-adsorbed liquid and washing fractions were collected.
concentrated and lyophilized. whereby 12 g of a viscous substance was obtained.
This viscous substance was dissolved in 200 ml of distilled water. After adjustment to pH 3.3 with trifluoroacetic acid. the resulting solution was then charged on a column (1 L) equilibrated with 0.04% aqueous trifluoroacetic acid and packed with "Diaion CHP-20P" (product of Mitsubishi Chemical). After development of the column with 2 L of 0.04% aqueous trifluoroacetic acid and pooling of the fraction Doc: FP9907s4.doc P81485/FP-9907( PCT)/tsa-gad-sh/English translation of spec (pages 135-21 8Examples)/I 8.12.00 (Fraction H) eluted between 0.8 and 1.4 L. the eluting solution was changed to 2 L of distilled water. Concentration and lyophilization of 2 L of the fraction (Fraction F) eluted with distilled water yielded 605 mg of a powder.
600 ml of Fraction H was diluted with distilled water to 1 L and its pH adjusted to 2.8 with trifluoroacetic acid. and the resulting solution was then charged again on a column (1 L) packed with "Diaion CHP-20P" (product of Mitsubishi Chemical) equilibrated with 0.04% aqueous trifluoroacetic acid. The column was eluted with 2.2 L of 0.04% aqueous trifluoroacetic acid. Fractions 8 to 11 obtained by fractionation of the eluate in portions of 200 ml each were concentrated and lyophilized, whereby 233 mg of a powder was obtained.
A 100 mg portion of the resulting powder was dissolved in 5 ml of water and 1 ml portions of the resulting solution were charged on an HPLC column ("Senshu Pak ODS-H-5251": 204 x 250 mm: product of Senshu Scientific) equilibrated with 0.04% aqueous trifluoroacetic acid. The column was developed at a flow rate of 10 ml/min.
The ultraviolet absorption of the active fraction at 210 nm was detected and a peak eluted during a retention time of 14 to 16 minutes was collected. the process being carried out 5 times. The fractions thus obtained were concentrated by a rotary evaporator, followed by lyophilization. whereby 23 mg of Compound A-500359H was obtained as a pure product.
In 15 ml of water were dissolved 605 mg of lyophilized powder of Fraction F and 1 ml portions of the resulting solution were charged on an HPLC column ("Senshu Pak ODS-H-5251": 206 x 250 mm: product of Senshu Scientific) equilibrated with 0.04% aqueous trifluoroacetic acid. The column was developed at a flow rate of 10 ml/min. The absorption of the active fraction at the ultraviolet portion of 210 nm was detected and a peak eluted during a retention time of 29 to 31 minutes was collected 15 times by fractionation. The fractions thus obtained were concentrated by a rotary evaporator, followed by lyophilization. whereby 134 mg of Compound A-500359F was obtained as a pure product.
The compound A-500359F has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol, insoluble in normal hexane and chloroform 3) Molecular formula: C 17
HIN
3
O
1 2 Doc: FP9907s4.doc P81485/FP-9907( PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 200 4) Molecular weight: 459 (as measured by FAB mass spectrometry) Accurate mass, as measured by high-resolution FAB mass spectrometry is as follows: Found: 460.1201 Calculated: 460.1203 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 262 nm (E 7.000) 7) Optical rotation: optical rotation measured in water exhibits the following value: 111 (c 0.41) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3391.2941. 1684. 1466. 1400. 1333. 1269. 1205. 1137. 1115. 1062. 1020 cm'.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 3.37 (3H. 3.79 (1H. dd. J=5.1. 6.4Hz). 4.17 (1H. ddd. J=1.6. 3.4. 4.6 Hz). 4.38 (1H.
dd. J=3.5. 5.1 Hz). 4.48 (1H. dd. J=2.4. 6.4 Hz). 4.49 (1H. ddd. J=0.6. 2.7. 4.6 Hz).
4.69 (1H. d. J=2.4 Hz). 5.32 (1H. dd. J=0.6. 3.4 Hz), 5.77 (1H. d. J=3.5 Hz). 5.90 (1H, d. J=8.1 Hz). 6.11 (1H. dd. J=1.6. 2.7 Hz). 7.75 (1H. d. J=8.1 Hz) ppm.
'C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1.4-dioxane (67.4 ppm) as an internal standard substance. 'C nuclear magnetic resonance spectrum is as follows: 58.6 62.7 65.5 72.7 76.3 78.8 91.2 100.0 102.7 114.8 140.7 141.9 152.1 165.4 167.0 173.9 ppm.
11) HPLC analysis: Column: "Senshu Pak ODS-H-2151" 6( x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 ml/min Detection: UV 210 nm Retention time: 18 minutes Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/1 8.12.00 Compound A-500359H has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol, insoluble in normal hexane and chloroform 3) Molecular formula: C 1 6
H
19
N
3 0 12 4) Molecular weight: 445 Accurate mass, as measured by high-resolution FAB spectrometry is as follows: Found: 446.1025 Calculated: 446.1047 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 262 nm (e 7.400) 7) Optical rotation: optical rotation measured in water exhibits the following value: +1150 (c 0.33) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3361. 2934. 1683. 1467. 1403. 1336. 1270. 1206. 1114. 1090. 1058. 1021 cm'.
9) 1H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 4.13 (br. t. J=5.4 Hz). 4.15-4.19 4.43 (1H. dd. J=2.5. 5.8Hz). 4.48 (1H. dd.
J=2.9. 4.7 Hz). 4.72 (1H. d. J=2.5 Hz). 5.31 (1H. d. J=4.0 Hz). 5.80 (1H. d. J=4.0 Hz), 5.89 (1H. d. J=8.3 Hz). 6.12 (1 H. dd. J= 1.4. 2.9 Hz). 7.75 (1H. d. J=8.3 Hz) ppm.
'1C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1.4-dioxane (67.4ppm) as an internal standard substance. 3C nuclear magnetic resonance spectrum is as follows: 62.8 65.8 70.3 74.6 77.0 84.2 90.3 100.3 102.9 113.9 141.2 141.9 152.2 165.9 167.0 174.2 ppm.
11) HPLC analysis: Column: "Senshu Pak ODS-H-2151" 6( x 150 mm (product of Senshu Scientific Co.. Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 ml/min Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-ead-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 202 Detection: UV 210 nm Retention time: 8 minutes Example 46: Cultivation of Streptomyces griseus Strain SANK 60196 (FERM BP-5420) Into each of three 2 L Erlenmeyer flasks, each containing 500 ml of the seed culture medium having the composition described below were aseptically inoculated four loopfuls of Strain SANK60196. These flasks were shaken on a rotary shaker at 23 0 C and 210 rpm and thus, the initial seed culture was conducted for 3 days.
The seed culture medium contains the following components in 1000 ml of tap water.
Glucose 20 g Soluble starch 10 g Pressed yeast 9 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g Antifoamer "CB442" 50 mg (product of NOF Corporation) After adjustment of pH to 7.4, sterilization was conducted at 121°C for minutes.
The first seed culture thus obtained was inoculated at 3% into a 60 L tank containing 30 L of the same preculture medium, and the second seed culture was carried :out with aeration and agitation at 23 0 C for 24 hours.
Cultivation was conducted as described below. Described specifically, the second seed culture broth was inoculated at 3% into two 600 L tanks, each containing 400 L of the below-described cultivation medium and cultivation was then carried out with Saeration and agitation at 23 0 C for 6 days.
The medium for cultivation: containing the following components in 1000 ml of tap water.
Glucose 20 g Soluble starch 10 g Doc: FP9907al.doc P81485/FP-9907(PCTtzsa/gadshcorcted pages of spcdO3/01/01 Pressed veast 9 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g Antifoamer "CB442" 50 mg (product of NOF Corporation) After adjustment to pH 7.4. 3 g of calcium carbonate was added and the mixture was sterilized at 125-C for 20 minutes.
Example 47: Purification of Compound A-500359E The cultured broth (810 L) obtained in Example 46 was filtered with the aid of "Celite 545" (product of Celite Corporation).
Upon subsequent purification, the active fraction was monitored by HPLC using the following column and analytical conditions.
Column: "YMC-Pak ODS-A A-312" 64 x 150 mm (product of YMC) Solvent: 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile Flow rate: 1.0 ml/min Detection: UV 210 nm Retention time: 19.8 minutes The resulting filtrate (800 L) was charged on a column (160 L) packed with "Diaion HP-20P" (product of litsubishi Chemical). The column was washed with 640 L of deionised water and the non-adsorbed fraction and washing fraction were then combined (non-adsorbed-washing fraction). The adsorbed substance was eluted with 348 L of 10% aqueous acetone.
After concentration of the eluted fraction to 10 L. the residue was charged on a column (45 L) packed with "Diaion CHP-20P" (product of Mitsubishi Chemical).
The column was then washed with 90 L ofdeionised water. 100 L of 10% aqueous methanol and 100 L of 15% aqueous methanol. The adsorbed substance was eluted with 100 L of 20% aqueous methanol.
After concentration of the 20% aqueous methanol fraction to 5 L. the concentrate was charged on a column (22 L) packed with "Toyopearl Doc: FP9907s4.doc P81485/FP-9907 PCT)/tsa-gad-sh/English translation of spec (pages 135-21 8:Examples)/l 8.12.00 (product of TOSOH Corporation). The column was developed with deionised water and the eluate was collected by fractionation in portions of 5 L each. The active substance having a retention time of 19.8 minutes upon the above-described HPLC was eluted in Fractions Nos. 3 to 6. These fractions were concentrated to 5.8 L and lyophilized to yield 55.8 g of a powder.
The resulting powder was dissolved in 1.2 L of deionised water. A 200 ml portion of the resulting solution was charged on an HPLC column ("YMC-Pak ODS- 1050-20-SR": 100p x 500 mm: product of YMC) equilibrated with 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile. The column was developed at a flow rate of 200 ml/min with 0.04% aqueous trifluoroacetic acid containing 4% acetonitrile. The active substance had a retention time of 105 to 124 minutes. That operation was repeated 6 times. The fractions thus obtained were combined.
concentrated to 5 L by "Evapor" and then lyophilized. whereby 24.2 g of Compound A-500359E was obtained as a pure product.
Example 48: Purification of Compounds A-500359F and A-500359H Upon subsequent purification, the active fraction was monitored by HPLC using the following column and analytical conditions.
Column: "YMC-Pak ODS-A A-312" 60 x 150 mm (product of YMC) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 mlmin Detection: UV 210 nm Retention time: 7.7 minutes (Compound A-500359H) 16.6 minutes (Compound A-500359F) The non-adsorbed-washing fraction (1370 L) obtained in Example 47 was charged on an activated charcoal column (65 After the column was washed with 260 L of deionised water. the active substance was eluted with 270 L of 0.5N aqueous ammonia containing 10% acetone. After concentration of the eluate to 40 L and adjustment of the concentrate to pH 2.4 with trifluoroacetic acid. it was charged on a column (45 L) packed with "Diaion CHP-20P" (product of Mitsubishi Chemical) equilibrated with 0.04% aqueous trifluoroacetic acid. The column was developed with 0.04% aqueous trifluoroacetic acid to yield a fraction (Fraction H) eluted in 0 to 47 L and another fraction (Fraction F) eluted in 47 to 91 L. Fraction H was Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218: Examples)/ 18.12.00 concentrated to 1.5 L. while Fraction F was obtained as 287 g of a powder after concentration and lyophilization.
The concentrate of Fraction H was diluted with deionised water to 3.2 L. A 160 ml portion of it was charged on an HPLC column ("YMC-Pack ODS-1050-20- SR": 100 x 500 mm: product of YMC) equilibrated with 0.04% aqueous trifluoroacetic acid. followed by development at a flow rate of 200 ml/min. Ultraviolet absorption of the active fraction at 210 nm was detected and a peak eluted at a retention time of 67 to 72 minutes was collected by fractionation. This operation was repeated 20 times. The fractions thus obtained were concentrated by "Evapor" (product of Okawara Seisakujo) and lyophilized to yield 5.9 g of Compound A- 500359H as a pure product.
A 277 g portion of Fraction F in powder form was dissolved in 50 L of deionised water and the resulting solution was adjusted to pH 2.2 with trifluoroacetic acid. The solution was charged again on a column (45 L) packed with "Diaion CHP- (product of Mitsubishi Chemical) equilibrated with 0.04% aqueous trifluoroacetic acid. After washing the column with 97 L of 0.04% aqueous trifluoroacetic acid, the active substance was eluted with 120 L of deionised water.
The deionised water eluted fraction was concentrated and lyophilized. whereby 75.6 g of Fraction F was obtained as a lyophilized powder.
The resulting lyophilized powder of Fraction F was dissolved in 4 L of water.
A 150 ml portion of the solution was charged on an HPLC column ("YMC-Pak ODS- 1050-20-SR". 100 x 500 mm: product of YMC) equilibrated with a mixture of acetonitrile and 0.04% aqueous trifluoroacetic acid. followed by development with the same solvent system at a flow rate of 200 ml/min. The absorption of the active fraction at the ultraviolet portion of 210 nm was detected and a peak eluted at a retention time of 88 to 97 minutes was collected by fractionation. This operation was repeated 27 times. The fractions thus obtained were concentrated and lyophilized.
whereby 19.2 g of Compound A-500359F was obtained as a pure product.
Example 49: Preparation process of each of Compound A-500359F and the amide derivative of compound A-500359F (chemical conversion of Compound A-500359E by aqueous ammonia) Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 Compound A-500359E (75 mg) obtained in Example 44 was dissolved in 2 ml of 0.5N aqueous ammonia. The resulting solution was allowed to stand at room temperature for 2 hours. After completion of the reaction, the reaction mixture was lyophilized to yield 78 mg of a powder.
The resulting powder was dissolved in 1 ml of 0.04% aqueous TFA. A 100 l portion of the resulting solution was charged on an HPLC column ("Capcellpak UG 120A". 200 x 250 mm; product of Shiseido) equilibrated with 0.04% aqueous trifluoroacetic acid, followed by elution with 0.04% aqueous trifluoroacetic acid at a flow rate of 10 ml/min. The ultraviolet absorption of the active fraction at 210 nm was detected and peaks eluted at a retention time of 21 to 22 minutes and at a retention time of 31 to 33 minutes were collected by fractionation. the process being carried out 10 times.
The fractions eluted at a retention time of 21 to 22 minutes were concentrated by a rotary evaporator and lyophilized. whereby 14 mg of the amide derivative of compound A-500359F was obtained in pure form.
The fractions eluted at a retention time of 31 to 33 minutes were concentrated by a rotary evaporator and lyophilized. wvhereby 50 mg of Compound A-500359F was obtained in pure form.
The amide derivative of compound A-500359F has the following physicochemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol. insoluble in normal hexane and chloroform 3) Molecular formula: C 7 H: NOII 4) Molecular weight: 458 (as measured by FAB mass spectrometry) Accurate mass. as measured by high-resolution FAB mass spectrometry is as follows: Found: 459.1328 Calculated: 459.1364 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 258 nm (E 7,500) 7) Optical rotation: optical rotation measured in water exhibits the following value: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/l 8.12.00 [aC]D2: -119' (c 0.87) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3339. 2943. 1686. 1598. 1495. 1402. 1337. 1272. 1205. 1136. 1115. 1060. 1019 cm-.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the signal of water as 4.75 ppm. 'H nuclear magnetic resonance spectrum is as follows: 3.30 (3H, s) 3.67 (1H. dd. J=5.0. 6.8 Hz). 4.17 (1H. ddd, J=1.8. 2.9. 4.4 Hz). 4.35 (1H.
dd, J=3.2. 5.0 Hz), 4.43 (1H. dd. J=2.3. 6.8 Hz). 4.45 (1H. dd. J=2.4. 4.4 Hz). 4.66 (IH. d. J=2.3 Hz). 5.35 (1H. d. J=2.9 Hz). 5.71 (1H, d. J=3.2 Hz). 5.85 (1H. d. J=8.1 Hz). 5.97 (1H. dd, J=1.8. 2.4 Hz). 7.71 (1H. d. J=8.1 Hz) ppm.
'1C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1.4-dioxane (67.4 ppm) as an internal standard substance. '1C nuclear magnetic resonance spectrum is as follows: 58.6 62.7 65.3 72.6 75.7 78.7 82.3 91.3 99.8 102.7 110.8 141.9 142.3 152.1 166.0 167.0 ppm.
11) HPLC analysis: Column: "Senshu Pack ODS-H-2151". 64 x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.5 ml/min Detection: UV 210 nm Retention time: 11 minutes Example 50: Preparation of Compound A-500359F (hydrolysis of Compound A- 500359E by sodium hydroxide) Compound A-500359E (4.4 mg) obtained in Example 44 was dissolved in ml of distilled water. After the dropwise addition of 0.5 ml of 0.02N aqueous sodium hydroxide, 1 ml of 0.1N aqueous sodium hydroxide was added dropwise. The resulting mixture was allowed to stand at room temperature for 50 minutes. The reaction mixture was neutralized with IN hydrochloric acid and then charged on 2 ml of an activated charcoal column. The column was washed with 8 ml of distilled water and the reaction substance was then eluted with 8 ml of 0.5N aqueous ammonia containing 10% acetone.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 208 After concentration of the eluate to 700 4l. the concentrate was charged on an HPLC column ("Senshu Pak ODS-H-4251": 10 x 250 mm; product of Senshu Scientific) equilibrated with 0.04% aqueous trifluoroacetic acid. followed by elution at a flow rate of 4 ml/min. The ultraviolet absorption of the active substance at 210 nm was detected and a peak eluted at a retention time of 25 to 30 minutes was collected by fractionation. This operation was repeated three times. The fractions thus obtained were concentrated in a rotary evaporator and lyophilized. whereby 2.6 mg of Compound A-500359F was obtained in pure form.
Example 51: Cultivation of Strepromy.ces griseus Strain SANK601 96 (FERM BP- 5420) One loopful of strain SANK60196 was sterilised before being inoculated into a 500 ml Erlenmever flask (seed flask) containing 100 ml of a medium having the composition described below. Seed culture was conducted for 3 days by shaking the flask in a rotary shaker at 23 C and 210 rpm.
Seed culture medium containing the following components in 1000 ml of tap water.
Maltose 30 g Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g "Antifoamer CB442" 50 mg After adjustment to pH 7.4. sterilization was conducted at 121 "C for 30 minutes.
Cultivation was conducted as described below. Described specifically, the seed culture was inoculated at 300 into each of ten 500 ml Erlenmever flasks.
each containing 100 ml of a sterilized medium having the composition described below. Cultivation was conducted for 11 days by shaking the flasks in a rotary shaker at 23 0 C and 210 rpm.
Cultivation medium: containing the following components in 1000 ml of tap water.
Glucose 50 g Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-21 8:Examples)/18.12.00 Meat extract 4 g Polypeptone 3 g Skimmed milk 10 g Corn steep liquor 10 g Sodium chloride 5 g "Antifoamer CB442" 50 mg After adjustment to pH 7.4. sterilization was conducted at 125°C for 30 minutes.
Example 52: Purification of Compound A-500359J Upon subsequent purification, the active fraction was monitored by HPLC using the following column and analytical conditions.
Column: "Pegasil ODS". 6) x 150 mm (product of Senshu Scientific Co..
Ltd.) Solvent: 0.04% aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 260 nm Retention time: 5.57 minutes The cultured broth obtained in Example 51 was filtered with the aid of"Celite 545" added at 5% The filtrate (1 L) thus obtained was charged on a column (200 ml) of"Diaion HP-20". The column was then washed with distilled water (500 ml).
After adjustment of the pH of 1.5 L of non-adsorbed-washing fraction to 9 with 6N sodium hydroxide, the fraction was charged on a column (100 ml) of"Dowex SBR-P The column was washed with distilled water (300 ml) and the adsorbed substance was eiuted with 300 ml of IN aqueous hydrochloric acid.
After adjustment of pH after elution to 7 with sodium hydroxide. the eluate was charged on an active charcoal column (50 ml). The column was washed with distilled water (100 ml) and the active substance was diluted with 60% aqueous acetone (200 ml). Concentration and lyophilization of the eluate yielded 558 mg of a powder.
The powder was dissolved in 5 ml of distilled water and 500 il portions of the resulting solution were charged on an HPLC column ("Senshu Pack Pegasil ODS": x 250 mm; product of Senshu Scientific) equilibrated with 0.05% aqueous trifluoroacetic acid. They were developed at a flow rate of 10.0 ml/min. The ultraviolet absorption of the active substance at 260 nm was detected and a peak Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/I 8.12.00 210 eluted at a retention time of 11.1 minutes was collected by fractionation, the process being carried out 10 times. The resulting fractions were concentrated by a rotary evaporator and then lyophilized. whereby 16.2 mg of Substance A-500359J was obtained in pure form.
The compound A-500359J has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water. slightly soluble in methanol, insoluble in normal hexane and chloroform 3) Molecular formula: Ci 6
HIN
3 0 1 3 4) Molecular weight: 463 (as measured by FAB mass spectrometry) Accurate mass. as measured by high-resolution FAB mass spectrometry is as follows: Found: 462.0996 Calculated: 462.1006 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 194 (e 8800). 262 (E 10000) nm 7) Optical rotation: optical rotation measured in water exhibits the following value: [c]D 28 -83o (c 0.1. HO) 8) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3372. 2931. 1684. 1467. 1407. 1273. 1204. 1107. 1058 cm".
9) H nuclear magnetic resonance spectrum was measured in deuterium oxide with 1.4-dioxane (3.53 ppm) as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follow\s: 3.75 (1H. t. J=3.4 Hz). 3.83 (1H. ddd. J=1.4. 1.9. 3.4 Hz). 4.02 (1H. ddd. J=1.4. 1.7, 3.4 Hz). 4.05 (1H. dd. J=5.3. 5.6 Hz). 4.11 (1H. t. J=5.6 Hz). 4.13 (1H. dd. J=3.1. 5.6 Hz). 4.30 (1H. d. J=5.3 Hz). 4.33 (1H. d. Hz). 4.90 (1H. d. J=1.9 Hz). 5.50 (1H.
d. J=3.1 Hz). 5.7 (1H.d. J=8.2 Hz). 7.6 (1H. d. J=8.2 Hz) ppm.
3C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1,4-dioxane (67.4 ppm) as an internal standard substance. '1C nuclear magnetic resonance spectrum is as follows: Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-shEnglish translation of spec (pages 135-218:Examples)/l 8.12.00 64.4 68.8 68.9 69.7 71.4 73.0 75.4 82.8 90.7 99.2 101.7 141.6 151.0 165.9 171.9 172.6 ppm.
11) HPLC analysis: Column: "Senshu Pak ODS-H-2151". 64 x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 0.05% aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 260 nm Retention time: 5.57 minutes Example 53: Cultivation of Streptomces griseus Strain SANK 60196 (FERM BP- 5420) One loopful of strain SANK60196 was sterilised prior to inoculation in a 500 ml Erlenmever flask (seed flask) containing 100 ml of a medium having the composition described below. Preculture was conducted for 3 days by shaking the flask in a rotary shaker at 23'C and 210 rpm.
Medium for preculture: containing the following components in 1000 ml of tap water.
Maltose 30 a Meat extract 5 g Polypeptone Sodium chloride 5 g Calcium carbonate 3 e "Antifoamer CB442" 50 mg After adjustment to pH 7.4. sterilization was conducted at 121 C for minutes.
Cultivation was conducted as described belo\w. Described specifically, the preculture broth was inoculated at 3%o into each often 500 ml Erlenmeyer flasks. each containing 100 ml of a sterilized medium having the composition described below. Cultivation was conducted by shaking the flasks in a rotary shaker at 23°C and 210 rpm. Six hours after initiation of the cultivation, filter-sterilized S- (2-aminoethyl)-L-cysteine hydrochloride and L-allylglycine were added to give a final concentration of 10 mM. Cultivation was then continued for 7 days.
Doc: FP9907s4.doc P81485/FP-9907( PCT/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 -'1 Cultivation medium: containing the following components in 1000 ml of tap water.
Maltose 30 g Yeast extract 5 g (product of Difco Laboratories) Meat extract 5 g Polypeptone 5 g Sodium chloride 5 g Calcium carbonate 3 g "Antifoamer CB442" 50 mg After adjustment to pH 7.4. sterilization was conducted at 125 0 C for minutes.
Example 54: Purification of Substance A-500359M-3 The cultured broth (1 L) obtained in Example 53 was centrifuged at 3000 rpm for 20 minutes and the resulting supernatant was purified.
Upon subsequent purification, the active fraction was monitored by HPLC using the following column and analytical conditions.
Column: "Pegasil ODS" 64 x 150 mm (product of Senshu Scientific) Solvent: 7.2% acetonitrile 0.05% aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 260 nm Retention time: 10.1 minutes After adjustment of the supernatant to pH 3 with trifluoroacetic acid. the resulting solution (1 L) was charged on a "Diaion HP-20" column (200 ml) equilibrated with 0.05% aqueous trifluorocetic acid. The column was washed with 0.05% aqueous trifluoroacetic acid (500 ml). followed by elution with distilled water (500 ml). The distilled water eluate (500 ml) thus obtained was concentrated and lyophilized to yield 230 mg of a crude powdery product.
The crude powdery product was dissolved in 2 ml of distilled water and a 500 pl portion of the resulting solution was charged on an HPLC column ("Pegasil ODS".
trade name: 204 x 250 mm; product of Senshu Scientific) equilibrated with 0.05% aqueous trifluoroacetic acid containing 7% acetonitrile.
Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-sh/English translation of spec (pages 135-218:Examples)/18.12.00 213 The column was developed with the same solvent at a flow rate of 10.0 ml/min and the ultraviolet absorption at 210 nm was monitored, resulting in elution of the active substance at a retention time of 28.0 minutes. This operation was repeated four times and the eluates were combined, concentrated and lyophilized, whereby 11.1 mg of Substance A-500359M-3 was obtained in pure form.
The compound A-500359M-3 has the following physico-chemical properties: 1) Appearance of the substance: white powder 2) Solubility: soluble in water and methanol, insoluble in normal hexane and chloroform 3) Molecular formula: C22H 28
N
4 03 4) Molecular weight: 556 (as measured by FAB mass spectrometry) Accurate mass, [M+H] as measured by high-resolution FAB mass spectrometry is as follows: Found: 557.1754 Calculated: 557.1731 6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water exhibits the following maximum absorption: 236 nm (c 10,000) 7) Optical rotation: optical rotation measured in water exhibits the following value: S[aD 26 +920 (c 0.1, H 2 0) Infrared absorption spectrum: Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhibits the following absorption maxima: 3407, 2938, 1684, 1524, 1465, 1399, 1385, 1335,1268, 1205, 1139, 1118, 1095, 1063, 1021 cm'.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with 1,4dioxane (3.53 ppm) as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows: 2.44 (1H, ddd, J=4.3, 7.3, 13.3 Hz), 2.52 (1H, ddd, J=4.3, 7.5, 13.3 Hz), 3.27 (3H, s), 3.66 (IH, t, J=5.5 Hz), 4.17 (1H, ddd, J=1.l, 2.5, 3.1 Hz), 4.32 (1H, dd, J=3.7, 5.5 Hz), 4.33 (1H, t, J=4.3 Hz), 4.45 (1H, 4.46 (1H, 4.73 (1H overlapped with HDO), 5.07 (1H, d, J=10.2 Hz), 5.36 (1H, d, J=3.1 Hz), 5.51 (1H, d, J=17.1 Hz), 5.58 (1H, d, J=8.1 Hz), 5.73 (1H, 5.74 (1H, d, J=3.7 Hz), 5.95 (1H, dd, J=1.1, 1.9 Hz), 7.72 (1H, d, J=8.1 Hz) ppm.
Doc: FP9907al.doc P81 1485/FP-9907(PCT)/tsa/gad/sh/cofrcced pages of spedO3101/01 214 3C nuclear magnetic resonance spectrum was measured in deuterium oxide with 1,4dioxane (67.4 ppm) as an internal standard substance. 3 C nuclear magnetic resonance spectrum is as follows: 37.1 55.4 58.6 62.6 65.3 72.6 75.7 78.9 82.4(d), 90.6 99.8(d), 102.6 109.9 119.0 134.0 141.7 142.2 152.0 162.3 166.8 173.6 177.6 ppm.
11) HPLC analysis: Column: "Pegasil ODS" 6< x 150 mm (product of Senshu Scientific Co., Ltd.) Solvent: 7.2% acetonitrile 0.05% aqueous trifluoroacetic acid Flow rate: 1.0 ml/min Detection: UV 260 nm Retention time: 10.1 minutes Test 1. Antibacterial activity Minimum inhibitory concentration The minimum inhibitory concentration of the compounds of the invention against Mycobacterium smegmatis Strain SANK 75075 was determined in accordance with the process described below. The concentration of the compound to be tested was set at four stages by four-fold dilution starting from 1000 pg/ml (1000 pg/ml, 250 pg/ml, 62 pg/ml and 15 pg/ml). A I ml portion of the diluted sample of each stage was poured into a Petri dish ("Terumo Petri dish", 90 x 20 mm). A nutrient agar medium (9 ml, product of Eiken Chemical) containing 5% glycerol was added and they were mixed to prepare a plate medium. A test microorganism Mycobacterium smegmatis SANK 75075 was precultured overnight at 37°C on a trypto-soy broth medium (product of Eiken Chemical) containing 5% glycerol. On the testing day, the microorganism solution was diluted 100fold with T.S.B. and one loopful of the diluted culture was streaked onto the plate medium. After cultivation at 37 0 C for 18 hours, the minimum concentration (MIC) of the test substance inhibiting the growth of the microorganism was determined. The results are shown in Table 6.
Doc: FP9907al.doc P81 4 85/FP-9907(PCT/tsa/gad/sh/corrccted pages ofspcc/03/01/01 215 Table 6 Antibacterial activities against Mycobacterium smegmatis SANK 75075 Exemp. Compound No. Minimum inhibitory concentration (pg/ml) 1 6.2 7 6.2 8 9 3.1 6.2 11 6.2 16 6.2 17 6.2 18 3.1 3.1 51 52 3.1 53 135 282 6.2 548 6.2 891 6.2 1091 6.2 Capuramycin 12.5 The minimum inhibitory concentration of the invention compound of the formula (Ia) against Mycobacterium avium Strain NIHJ 1605 was determined. Described specifically, Tween 80 was added to Middleblook 7H9 broth. After autoclave S. sterilization, Middleblook ADC enrichment was added Into each of micro-test 0* tubes was poured a 0.8 ml portion of the resulting mixture. To each of the test tubes was added a 0.1 ml portion of each of the compounds of the invention diluted two-fold (which will hereinafter be abbreviated as "medicament-containing medium"). On the side, a colony obtained by preculturing Mycobacterium avium Doc: FP9907al.doc P8 485/FP-9907(PCT)tsa/gadshcorected pages of specJ03/01101 1- NIHJ1605 on a Tween egg medium for 10 to 14 days was charged in a test tube containing Tween 80 and glass beads. After sufficient mixing, Middleblook 7H9 broth was added to form a uniform microorganism solution. The microorganism solution was adjusted to OD 625 nm 0.10 (viable cell count: about 1 x 108 CFU/ml).
followed by 100-fold dilution. A 0.1 ml portion of the resulting microorganism solution was inoculated into the above-described medicament-containing medium (final viable cell count: about I x 10l CFU/ml). followed by aerobic culture at 37°C for 6 days. The minimum medicament amount at which no colony having a diameter of 1 mm or greater was recognized on the bottom of the test tube was determined as MIC (pg/ml). The results are shown in Table 7.
Table 7 Antibacterial activities against .\Acobacterium avium NIHJ 1605 Exemp. compound Minimum inhibitory concentration No. (jg/ml) 539 0.25 571 1 594 1 Capuramycin 8 Disk Assay So-called disk assay was conducted using 40 pg of a test substance per paper disk of 8 mm. Compound A-5003591-2 (Exemp. compound No. 396) exhibited an inhibitory zone of 14 mm in diameter against Bacillus suhiilis PCI 219. that of 30 mm in diameter against Mycobacterium smegmais SANK 75075 and that of 25 mm in diameter against Klebsiella pneumoniae PCI 602.
So-called Disk assay ("Experimental Agricultural Chemistry". ed. by Agricultural Chemistry Class/Agriculture Dept.!Tokyo Univ.. 3rd edition. Volume II.
published by Asakura Shoten in 1978) was conducted using 40 utg of a test substance per paper disk of 8 mm. Compound A-500359E exhibited an inhibitory circle of 12 mm in diameter against Mycobacterium smegmatis SANK 75075. the amide Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-ead-sh/English translation of spec (pages 135-218: Examples)/ 18.12.00 derivative of compound A-500359F exhibited an inhibitory circle of 12 mm in diameter and Compound A-500359M-3 also exhibited an inhibitory circle of 12 mm in diameter.
Preparation Example 1 Capsules A-500359A or C 100mg Lactose 100 mg Corn starch 148.8 mg Magnesium stearate 1.2 mg Total amount 350 mg A capsule was obtained by mixing powders in accordance with the abovedescribed formulation. sieving the resulting mixture through a 60-mesh sieve, and then charging the resulting powder in a gelatin capsule.
Preparation Example 2 Capsules were each obtained by mixing 100 mg of Compound A-500359E, Compound A-500359F. the amide derivative of compound A-500359F. Compound A-500359H. Compound A-500359J or Compound A-500359M-3. 100 mg of lactose respectively. 148.8 mg of corn starch and 1.2 mg of magnesium stearate (totally. 350 mg) in the powdery form. sieving the resulting mixture through a 60-mesh sieve and charging the powder in a gelatin capsule.
Toxicity Test The invention compound A-500359A exhibited no toxicity when intravenously administered to a mouse in an amount of 500 mg/kg.
The results described above show that the compounds of the invention represented by the formulae (XII). (XIII). (XIV), (XV) and (XVI) respectively, various derivatives of the compound represented by the formula and pharmacologically acceptable salts thereof exhibit excellent antibacterial activities against various bacteria including Mycobacteria so that they are useful in the prevention or treatment of infectious diseases caused by such bacteria. Streptomyces Doc: FP9907s4.doc P81485/FP-9907(PCT)/tsa-gad-shlEnglish translation of spec (pages 135-218.Examples /18.12.00 P:\OPER\Kbm\2544791 spec.doc-I 7/ )3 21.8 griseus SANK60196 (FERM BP-5420) is useful as a bacterium producing the compound represented by the formula (XII), (XIV), (XV) or (XVI). The compounds of the invention represented by the formulae (XIII), (XIV), (XV) or (XVI) are also useful as a starting material for the synthesis of a derivative for the preparation of a prevention or treatment of various infectious diseases by organic chemical or microbiological conversion.
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 of suggestion that that prior art forms part of the common general knowledge in Australia.
*oooo

Claims (19)

1. A compound A-500359E of formula (XI) or a salt thereof. O OH OCH 3 OH (XI) 220
2. A compound A-500359F of formula (XII) or a salt thereof. OH H 3 C (XII)
3. An amide derivative of compound A-500359F of formula (XIII) or a salt thereof. H 3 C1 (XIII) 221
4. A compound A-500359H of formula (XIV) or a salt thereof.
OH (XIV) A compound A-500359J of formula (XV) or a salt thereof. (XV) P:AOPER\Kbm\2544791 Spec.doc-17/m6/3 222
6. A compound A-500359M-3 of formula (XVI) or a salt thereof. (XVI)
7. A process for preparing a compound according to any one of claims 1, 2, 4 or 5 by a cultivation procedure which comprises i) cultivating isolated Streptomyces griseus (SANK60196; FERM BP-5420) and ii) isolating the compound from the cultivation products.
8. Isolated Streptomyces griseus (SANK60196; FERM BP-5420) when used for producing a compound according to claim 1, 2, 4 or
9. A pharmaceutical composition comprising an effective amount of a pharmacologically active compound together with a carrier or diluent therefor, wherein said pharmacologically active compound is a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof. 15
10. The use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of a bacterial infection. oO° P'OPER\Kb ,'544791 di\-.doc.-2(106)2 -223-
11. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof for use as a medicament.
12. An agent containing a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a bacterial infection.
13. A method for the treatment or prevention of a bacterial infection in a mammal, which comprises administering to a mammal in need of such treatment an effective amount of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.
14. A process for preparing a compound according to any one of claims 1, 2, 4 or by a cultivation procedure, substantially as hereinbefore described.
Isolated Streptomyces griseus (SANK60196; FERM BP-5420) when used for producing a compound according to claim 1, 2, 4 or 5, substantially as hereinbefore described.
16. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, substantially as hereinbefore described.
17. A pharmaceutical composition containing a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, substantially as hereinbefore described.
18. The use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, substantially as hereinbefore described.
19. An agent containing a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament, substantially as hereinbefore described. P:'OPER\Kbmin2544791 div.doc-2106/0)2 -224- comprises administering to a mammal in need of such treatment an effective amount of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, substantially as hereinbefore described. DATED this 21st day of June, 2002 Sankyo Company Limited By DAVIES COLLISON CAVE Patent Attorneys for the Applicants
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