Indian Patents
Title of Invention

"NEW ANTIMICROBIAL COMPOUNDS"
Abstract A compound of formula (I) or a pharmaceutically acceptable salt thereof: wherein: R1 is a methyl group, R2 is s methyl group, R4 is a hydroxy group , and X is a methylene group; R1 is a methyl group, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a methylene group; R1 is a methyl group, R2 is a methyl group, R4 is a hydrogen atom, and X is a methylene group; R1 is a hydrogen atom, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a methylene group; or R1 is a methyl group, R2 is a methyl group, R4 is a hydroxy group, and X is a sulfur atom.
Full Text 2
The present invention relates to a compound of formula (I), an ester or ether
compound of formula(Ia) and an N-alkylearbamoyl derivative compound of formula (II) which have excellent antibiotic activity or a pharmaceutically acceptable salt thereof
Tlte present invention is also a pharmaceutical composition comprising a compound described above as an active ingredient.
The present invention includes a use of a compound described above in order to prepare a medicament effective to treat or prevent bacterial infections.
The present invention includes a microorganism capable of producing a compound of formula (I),
[Background of the invention]
A -lactam antibiotic, an amino-glycoside, isoniazid or rifampicin has been conventionally used in treatment or prophylaxis of microbial infections including tebercule 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 (J.Antibiotics, 29, (8), 1047-1053 (1986)).

We found new compounds of formula (I), which do not show any cross resistance to conventional medicaments, in the cultivation products of a microorganism. We prepared the derivatives of compounds

3
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 of treat and prevent infection diseases including ones arising from bacteria resistant to the conventional antibiotics. Compounds of formula (I) is also useful starting materials for preparation of the compounds of the present invention having excellent antibiotic activity. [Disclosure of the invention]
The present invention iucludes a compound of formula (I)

(wherein
R1 is a methyl group, R2 is a methyl group, R4 is a hydroxy group, and X is a methylene group;
R1 is a methyl group, R2 is a hydrogen atom, R4 is a hydroxy group, X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R4 is a hydrogen atom, and X is a methylene group;
R1 is a hydrogen atom, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a methylene group; or
R1 is a methyl group, R2 is a methyl group, R4 is a hydroxy group, and X is a sulfur atom) or a pharmaceutically acceptable salt thereof, or
A pharmaceutically acceptable ester or ether compound of formula (la) or N-alkylcarbamoyl derivative compound of formula (Ik)


-4-

wherein:
R1 is a hydrogen atom or a methyl group;
R2 is a hydrogen atom or a methyl group;
R3 is a hydrogen atom, a C6-20-alkylcarbonyI group, a C6-20-alkyloxycarbonyl group, a C10-20-alkenylcarbonyl group having 1 to 3 double bonds, or a C6-20-alkyl group;
R4 a is a hydrogen atom or a hydroxy group;
R5 is a hydrogen atom, a C6-20-alkylcarbonyl group, a C6-20-alkyloxycarbonyl group, or a C10-20-alkenylcarbonyl group having 1 to 3 double bonds;
R11 is a C1-21-alkyl group; and
X is a methylene group or a sulfur atom,
PROVIDED THAT:
in formula (Ia) one or two ester residues are present as one or two of -OR3 and -OR5, or one ether residue is present as -OR3, or a combination thereof; AND THAT: when X is a sulfur atom,
R1 is a methyl group, R2a is a methyl group and R4n is a hydroxy group; when X is a methylene group, R1 is a methyl group and R2a is a hydrogen atom, R4a is a hydroxy group; or when X is a methylene group and R1 is a hydrogen atom,
R2a is a methyl group, and R4a is a hydroxy group; or a pharmaceutically acceptable salt thereof.
The present invention is also a pharmaceutical composition comprising a compound described above as an active ingredient.

The present invention includes the use of a compound described above in order to prepare a medicament effective to treat or prevent bacterial infections.
The present invention includes a microorganism Streptomyces grisens SANK 60196 (FERM BP-5420) capable of producing a compound of formula (I).
The pharmaceutically acceptable ester, ether or N-alkylcarbamoyl derivative compound refers to a compound that is a useful medicament without significant toxicity.
When the compound of formula (Ia) is an ester compound, one or both of R3 and R5 is a carbonyl or oxycarbony] group to which a straight or branched chain C6 - C20 alkyl group is attached, in which said alkyl group is selected from the group consisting of hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimelhylbutyl, 1,2- o dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 1-propylbuty], 4,4-dimethylpentyl, octyl, 1-methylbeptyl, 2-methylheptyl, 3-methylheptyl, 4-methylheptyl, 5-methylheptyl, 6-methylheptyl, 1-propylpentyl, 2-ethylhexyl, 5,5-dimethylhexyl, nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6-methyloctyl, 1-propylhexyl, 2-ethylheptyl, 6,6-dimethylheptyl, decyl, 1-methybonyl, 3-methyhionyl, 8-methylnonyl, 3-ethyloctyl, 3,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8-dimethylnonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,11-trimethyldodecyl, hexadecyl, 4,8,12-trimethyltridecyl,
1-methylpentadecyl, 14-methylpentadecyl, 13,13-dimethyltetradecy), heptadecyl, 15-methylhexadecyl, octadecyl, 1-methylheptadecyl, nonadecyl, icosyl and 3,7,11,15-tetramethylhexadecyl groups; or is a
carbonyl group to which a straight or branched chain C10- C20 alkenyl group is attached, in which said alkenyl group is selected from the group consisting of cis-8-heptadecenyl, cis, cis-8,11-heptadecadienyl, cis, cis, cis-8,ll,14-heptadecatrienyl and cis-10-nonadecenyl.
When the compound of formula (la) is an ether compound, R3 is a straight or branched chain C6 - C20 alkyl group such as the hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-

- 6-
dimethylbutyl, 2-ethylbutyI, heptyl, 1-methylhexy], 2-melhylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 1-propylbutyl, 4,4-dimethylpentyl, octyl, 1-methylheptyl, 2-methylheptyl, 3-methylheptyl, 4-methylheptyl, 5-methylheptyl, 6-methylheptyl, 1-propylpentyl, 2-ethylhexyl, 5,5-dimethylhexyl, nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6-methyloctyl, 1-propylhexyl, 2-ethylheptyl, 6,6-dimethyihepty], decyl, l-methylnonyl, 3-methylnonyj, 8-methyinonyl, 3-ethyloctyl, 3,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8-dimethylnonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,11-trimethyldodecyl, hexadecyl, 4,8,12-trimethyItridecy!, 1-methylpentadecyl, 14-methylpentadecyl, 13,13-dimethyltetradecyl, heptadecyl, 15-methylhexadecyl, octadecyl, 1-methylheptadecyl, nonadecyl, icosyl and 3,7,11,15-tetramethylhexadecyl groups:
The alkyl residue Rl1 of the N-alkylcarbamoyl derivative compound of formula (Ik) is selected from the group consisting of
. "straight or branched chain C1 - C21 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, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 1-propylbutyl, 4,4-dimethylpentyl, octyl, 1-methylheptyl, 2-methylheptyl, 3-methylheptyl, 4-methylheptyl, 5-methylheptyl, 6-methylheptyl, 1-propylpentyl, 2-ethylhexyl, 5,5-dimethylhexyl, nonyl, 3-methyloctyl, 4-methyloctyl, 5-methyloctyl, 6-methyloctyl, 1-propylhexyl, 2-ethylheptyl, 6,6-dimethylheptyl, decyl, 1-methyhionyl, 3-methylnonyl, 8-methylnonyl, 3-ethyloctyl, 3,7-dimethyloctyl, 7,7-dimethyloctyl, undecyl, 4,8-dimethyinonyl, dodecyl, tridecyl, tetradecyl, pentadecyl, 3,7,11-trimethyldodecyl, hexadecyl, 4,8,12-trimethyltridecyl, 1 -methylpentadecyl, 14-methylpentadecyl, 13,13-dimethyltetradecyl, heptadecyl, 15-methylhexadecyl, octadecyl, l-methylheptadecyl, nonadecyl, icosyl, 3,7,11,15-tetramethylhexadecyl and henicosyl groups.

A most preferable alkyl residue R11 of the N-alkylcarbamoyl derivative compound is a C6 - C20 alkyl group.
The pharmaceutically acceptable ester compound of (la) is a compound which has one or two of the ester residues at R3 and/or R5. A more preferable ester compound is a compound which has one ester residue at R3 or R5. A most preferable ester compound is a compound which has one ester residue at R3.
The pharmaccutically acceptable ether compound of (la) is a compound which has one ether residue at R3.
The pharmaceutically acceptable N-alkylcarbamoyl derivative compound of (Ik) is a compound having one alkyl residue at R11.
The term "pharmaceutically acceptable salt" refers to a salt that is a useful medicament without significant toxicity.
Where compound (I), pharmaceutically acceptable ester and ether compounds (Ia) and N-alkyl derivative compound (Ik) have a basic group such as an amino group, these compounds can be converted 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 (I), pharmaceutically acceptable ester, and ether compounds (Ia) and N-alkyl derivative compound (Dc) 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 (I) and pharmaceutically acceptable ester, ether and N-alkylcarbamoyl derivative compounds (la) and (Ik) are allowed to stand in the atmosphere, these compounds may take up water to form a hydrate. The present invention includes such hydrates. Compound (I) and pharmaceutically acceptable

ester, ether and N-alkylcarbamoyl derivative compounds (Ia) and (Ik) may absorb a solvent to form a solvate. The present invention includes such solvates..
Compound (I) and pharmaceutically acceptable ester, ether and N-alkylcarbamoyl derivative compounds (Ia) and (Ik) 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:

A preferable compound (I) is selected from the following compounds:
(1) a compound (I) wherein R2 is a methyl group,
(2) a compound (I) wherein R4 is a hydroxy group,
(3) a compound (I) wherein X is a methylene group;
or a compound wherein R2, R4 and X is selected in optional combination of (1), (2) and (3), for example:
(4) a compound (I) wherein R4 is a hydroxy group and X is a methylene
group, and
(5) a compound (I) wherein R2 is a methyl group, R4 is a hydroxy
group and X is a methylene group.

The following Tables 1 and 2 are intended to illustrate typical compounds (I) and (Ia) of the present invention and are not intended to limit the scope of this invention.



Table 1

10 -

11


12







13



14


15


16


- 17-


18


19


20


21


-22-


- 23-


24


-25-


26


27





Table 2

28

29


30







31


32


33


-34


-35-


36


37


38


39


40


41


42


43


-44-

In Tables 1 and 2
Exemp. comp. No. is exemplification compound number,
CH2 is methylene group,
Me is methyl group,
OH is hydroxy group,
A7 is heptanoyl group,
A8 is octanoyl group,
A9 is nonanoyl group,
A.l 0 is decanoyl group,
A12 is lauroyl group,
A14 is myristoyl group,
A15 is pentadecanoyl group,
A16 is palraitoyl group,
A17 is heptadecanoyl group,
A18 is stearoyl group,
A20 is arachidoyl group,
AO7 is heptanoyloxy group,
AO8 is octanoyloxy group,
AO9 is nonanoyloxy group,
AO10 is decanoyloxy group,
AO12 is lauroyloxy group,
AO14 is myristoyloxy group,
AO15 is pentadecanoyloxy group,
AO16 is palmitoyloxy group,
AO17 is heptadecanoyloxy group,
AO18 is stearoyloxy group,
AO20 is arachidoyloxy group,
AO22 is behenoyloxy group,
OLE is oleoyl group,
LE is linoleoyl group,

LEN is linolenoyl group, CES is cis-11-eicosenoyl group, MO is 2-methyloctanoyl group, MD is 2-methyldecanoyl group, MDD is 2-methyldodecanoyl group, MTD is 2-methyl tetradecanoyl group, MHD is 2-methylhexadecanoyi group, DMO is 2,2-dimethyloctanoyl group, DMD is 2,2-dimethyldecanoyl group, DMDD is 2,2-dimethyldodecanoyl group, DMTD is 2,2-dimethyltetradecanoyl group, DMHD is 2,2-dimethylhexadecanoyl group, C2 is ethyl group,
C3 is propyl group,
C4 is butyl group,
C5 is pentyl group,
C6 is hexyl group,
C7 is heptyl group,
C8 is octyl group,
C9 is nonyl group,
C10 is decyl group,
C11 is undecyl group,
C12 is dodecyl group,
C13 is tridecyl group,
C14 is tetradecyl group,
C15 is pentadecyl group,
C16 is hexadecyl group,
C6OC is hexyloxycarbonyl group,
C7OC is heptyloxycarbonyl group,
C8OC is octyloxycarbonyl group,
C9OC is nonyloxycarbonyl group,
C100C is decyloxycarbonyl group,
CHOC is undecyloxycarbonyl group,
C12OC is dodecyloxycarbonyl group,
MMA10 is 2-methyldecanoyl group,

45

46
MMA12 is 2-methyldodecanoyl group, MMA14 is 2-methyltetradecanoyl group, DMA10 is 2,2-dimethyldecanoyl group, DMA12 is 2,2-dimethyldodecanoyl group, DMA34 is 2,2-dimethyltetradecanoyl group.
In a compound of formula (Ib):
the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R a is a hydroxy group, R5 a is a hydrogen atom and X is a methylene group represents A-500359A (exemplification compound No. 1);
the compound wherein R1 is a methyl group, R2 is a hydrogen atom, R3B is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group represents A-500359C (exemplification compound No. 2);
the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydrogen atom, R5a is a hydrogen atom and X is a methylene group represents A-500359D (exemplification compound No. 3);
the compound wherein R1 is a hydrogen atom, R2 is a hydrogen atom, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group represents A-500359G (exemplification compound No. 45); and
the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a 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 exemplification compound No. (exemp. comp. No.) 1 to 254, 309 to 312, 338 to 341, 367 to 370, 396 to 478, 508 to 513, 537 to 588, 592 to 704, 708 to 820, 891 to 910, 914 to 990, 1091 to 1160,1164 to 1210, 1214 to 1240, 1341 to 1390, 1394 to 1401 and 1405 to 1412;
more preferable compounds include compounds of exemplification compound No. 1 to 3, 7 to 11, 45, 49 to 53, 90 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

47
908; 922 to 940, 1091 to 1108, 1122 to 1158, 1172 to 1190, 1341 to 1358 and 1372 to 1.390;
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;

that is
exemp.comp.No.l represents the compound wherein R1 is a methyl group, R2 is a methyl group, R a is a hydrogen atom, R a is a hydroxy group, R5 a is a hydrogen atom and X is a methylene group;
exemp.comp.No.2 represents the compound wherein R1 is a methyl group, R2 is a hydrogen atom, R3a is a hydrogen atom, R4a is a hydroxy group, R5B is a hydrogen atom and X is a methylene group;
exemp.comp.No.3 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydrogen atom, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.7 represents the compound wherein R is a methyl group, R is a methyl group, R3a is a decanoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.8 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a lauroyl group, R4B is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
atom and X is a methylene group;
exemp.comp.No.10 represents the compound wherein R1 is a methyl group, R2 is methyl group, R3a is a pentadecanoyl group, R4a is a hydroxy group, R5a is a hydrogi atom and X is a methylene group;
exemp.comp.No.l 1 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a palmitoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.9 represents the compound wherein R1 is a methyl group, R is methyl group, R3a is a myristoyl group, R4a is a hydroxy group, R58 is a hydrogen atom and X is a methylene group;
a oen

exemp.comp.No.45 represents, the compound wherein R is a hydrogen atom, R is a hydrogen atom, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.49 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a decanoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.50 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a lauroyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.51 represents the compound wherein R is a hydrogen atom, R is a methyl group, R38 is a myristoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.52 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a pentadecanoyl group, R4a is a hydroxy group, R a is a hydrogen atom and X is a methylene group;
exemp.comp.No.53 represents the compound wherein R is a hydrogen atom, R is a methyl group, R3a is a pabnitoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exempxomp.No.90 represents the compound wherein Rl is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decanoyl group and X is a methylene group;
exemp.comp.No.91 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a lauroyl group and X is a methylene group;
exemp.comp.No.92 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a myristoyl group and X is a methylene group;
exemp.comp.No.93 represents the compound wherein Rl is a methyl group, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a pentadecanoyl group and X is a methylene group;
exemp.comp.No.94 represents the compound wherein R is a methyl group, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a palmitoyl group and X is a methylene group;

exemp.comp.No.131 represents the compound wherein R1 is a hydrogen atom, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decanoyl group and X is a methylene group;
exemp.comp.No.132 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a lauroyl group and X is a methylene group;
exemp.comp.No.133 represents the compound wherein R is a hydrogen atom, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a myristoyl group and X is a methylene group;
exemp.comp.No.134 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5 a is a pentadecanoyl group and X is a methylene group;
exemp.comp.No.135 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a palmitoyl group and X is a methylene group;
exemp.comp.No.537 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hexyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.538 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a heptyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.539 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is an octyloxycarbonyl group, R4a is a hydroxy group, R5a 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, R3a is a nonyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.541 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a decyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.542 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is an undecyloxycarbonyl group, R4a is a hydroxy group, R5 a is a hydrogen atom and X is a methylene group;

50
exempxomp.No.543 represents the compound wherein R is a methyl group, R2 is a methyl group, R3a is a dodecyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.550 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hexyloxycarbonyl group and X is a methylene group;
exemp.comp.No.551 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a heptyloxycarbonyl group and X is a methylene group;
exemp.comp.No.552 represents the compound wherein R is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is an octyloxycarbonyl group and X is a methylene group;
exemp.comp.No.553 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a nonyloxycarbonyl 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, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decyloxycarbonyl group and X is a methylene group;
exemp.comp.No.555 represents the compound wherein R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is an undecyloxycarbonyl group and X is a methylene group;
exernp.comp.No.556 represents the compound wherein R1 is a methyl group, R is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a dodecyloxycarbonyl group and X is a methylene group;
exemp.comp.No.563 represents the compound wherein R1 is a hydrogen atom, R is a methyl group, R3a is a hexyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.564 represents the compound wherein R1 is a hydrogen atom, R is a methyl group, R3a is a heptyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.565 represents the compound wherein R1 is a hydrogen atom, R is a methyl group, R3a is an octyloxycarbonyl group, R48 is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;

51
exemp.comp.No.566 represents the compound wherein R is a hydrogen atom, R is a methyl group, R3a is a nonyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.567 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3B is a decyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.568 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is an undecyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.569 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a dodecyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.576 represents the compound wherein R is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hexyloxycarbonyl group and X is a methylene group;
exemp.comp.No.577 represents the compound wherein Rl is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a heptyloxycarbonyl group and X is a methylene group;
exemp.comp.No.578 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R a is an octyloxycarbonyl group and X is a methylene group;
exempxomp.No.579 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a nonyloxycarbonyl group and X is a methylene group;
exemp.comp.No.580 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decyloxycarbonyl group and X is a methylene group;
exemp.comp.No.581 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5 a is an undecyloxycarbonyl group and X is a methylene group;
exemp.comp.No.582 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a dodecyloxycarbonyl group and X is a methylene group;

exemp.comp.No.594 represents the compound wherein R1 is a methy] group, R2 is a methyl group, R3a is a decyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.710 represents the compound wherein R1 is a hydrogen atom, R2 is a methyl group, R38 is a decyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
exemp.comp.No.891 represents the compound wherein R1 is a methyl group, R11 is a methyl group, R3 is a hydrogen atom, and R5 is a hydrogen atom;
exemp.comp.No.895 represents the compound wherein R1 is a methyl group, R11 is a methyl group, R3 is a decanoyl group, and R5 is a hydrogen atom;
exemp.comp.No.925 represents the compound wherein R1 is a methyl group, R11 is a methyl group, R3 is a hydrogen atom, and R5 is a decanoyl group;
exemp.comp.No.1091 represents the compound wherein R1 is a methyl group, R11 is a dodecyl group, R3 is a hydrogen atom, and R5 is a hydrogen atom;
exemp.comp.No.1141 represents the compound wherein Rl is a hydrogen atom, R11 is a methyl group, R3 is a hydrogen atom, and R5 is a hydrogen atom;
exemp.comp.No.l 145 represents the compound wherein R is a hydrogen atom, R11 is a methyl group, R is a decanoyl group, and R is a hydrogen atom;
exemp.comp.No.l 175 represents the compound wherein R1 is a hydrogen atom, R11 is a methyl group, R3 is a hydrogen atom, and R5 is a decanoyl group; and
exemp.comp.No.1341 represents the compound wherein R1 is a hydrogen atom, R11 is a dodecyl group, R3 is a hydrogen atom, and R5 is a hydrogen atom.

53
Compounds of the present invention represented by the formula (I) or (Ia) can be prepared by the process as described below.
Compounds A-500359A (Exemp. compound No. 1), A-500359C (Exemp. compound No. 2), A-500359D (Exemp. compound No. 3), A-500359G (Exemp. compound No. 45) and A-500359M-2 (Exemp. compound No. 396) of the present invention each represented by the formula (I) are available by culturing a microorganism capable of producing the above described compounds, belonging to the Streptomyces spp. on a suitable medium and then recovering the compound from the cultured broth. Streptomyces 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/Ibaraki-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°C for 14 days on a medium specified by International Streptomyces Project (which will hereinafter be abbreviated as "ISP") [refer to Shirling, E.B. and Gottlieb, D., "Int. J. Syst. Bacteriol. 16, 313-340 (1996)".] Observation through an optical microscope indicates that substrate mycelia of SANK60196 are favourably grown and branched and show yellowish grey, yellowish brown or pale olive colour, but unlike the strain belonging to Nocardia spp., does not show cleavage or zigzag extension. Aerial mycelia 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 mycelia. Formation of sporangia, axial division of aerial mycelia, cleavage of aerial mycelia and sclerotia are not recognized.

54
2) Growth characteristics on various culture media
Growth characteristics of Strain SANK60196 on an agar medium after cultivation at 28°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, yellowish brown (10YR 5/6)
AM: Abundantly formed, velvety, pale brown (2.5Y 8/2)
R: Yellowish brown (10YR 5/8)
SP: Yellowish brown (10YR 6/8) Oat meal - agar (ISP 3);
G: Excellent, flat, yellowish brown (2.5Y 6/6)
AM: Abundantly formed, velvety, 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, yellowish brown (2.5Y 6/4)
AM: Abundantly formed, velvety, yellowish grey (7.5Y 9/2)
R: Yellowish brown (2.5Y 6/4) Glycerin - asparagine agar (ISP 5)
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);

G: Excellent, flat, pale olive color (5Y 8/3)
AM: Slightly produced, velvety, yellowish grey (5Y.9/I)
R: Pale yellow (5Y 8/6)
SP: Not produced Tyrosine agar (ISP 7)
G: Good, flat, grayish yellow brown (2.5Y 5/4)
AM: Abundantly formed, velvety, light olive grey (7.5Y 8/2)
R: Yellowish brown (10YR 5/4)
SP: Grayish yellow brown (2.5Y 4/3) Sucrose - nitrate agax;
G: Not so good, flat, pale yellow (5Y 8/6)
AM: Abundantly formed, velvety, light olive grey (7.5Y 8/2)
R: Dark yellow (5Y 8/8)
SP: Pale yellow (5Y 9/6) Glucose - asparagine agar;
G: Good, flat, pale yellow (5Y 9/3)
AM: Not so good, velvety, yellowish grey (5Y 9/1)
R: Yellowish grey (7.5Y 9/3)
SP: Not produced Nutrient agar (product of Difco Laboratories)
G: Good, flat, pale yellowish brown (2.5Y 8/3)
AM: Good, velvety, yellowish grey (5Y 9/1)
R: Yellowish grey (5Y 9/4)
SP: Not produced Potato extract - carrot extract agar,
G: Not so good, flat, yellowish grey (7.5Y 9/2)
AM: Not so good, velvety, yellowish grey (5Y 9/2)
R: Yellowish grey (7.5Y 9/3)
SP: Yellowish grey (7.5Y 9/3) Water agar;
G: Not good, flat, yellowish grey (5Y 9/1)
AM: Not good, velvety, yellowish grey (5ZY 9/1)
R: Yellowish grey (7.5Y 9/4)
SP: Pale yellow (5Y 9/6)

56
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 1 is a
yeast extract - malt extract agar medium (ISP 2).
[Table 4]
Hydrolysis of starch positive
Liquefaction of gelatin positive
Reduction of nitrates positive
Coagulation of milk negative
Peptonization of milk positive
Formation of melamine-like pigment positive
Substrate decomposition: casein positive
tyrosine positive
xanthine negative
Growth temperature range (Medium 1) 6 to 35°C
Optimum growth temperature (Medium 1) 18 to 30°C
Growth in the presence of salt (Medium I) 10%
Utilisation of a carbon source by Strain SANK60196 observed after cultivation at 28°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 non-uti Usable. [Table 5]
D-glucose +
L-arabinose
D-xylose +
Inositol
D-mannitol +
D-fructose +
L-rhamnose
Sucrose
Raffinose
Control

4) Chemotaxonomic properties
The cell wall of the present strain was investigated in accordance with the method of Hasegawa, et al. [refer to Hasegawa, T., et al., "The Journal of General and Applied Microbiology, 29, 319-322(1983)], resulting in the detection of LL-diaminopimelic 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, M.P., "Journal of Laboratory and Clinical Medicine, 71, 934-944(1968)]. As a result, no characteristic component was detected.
The above-described mycological properties have revealed that the present strain belongs to Streptomyces spp. among the actinomycetes. It has been made clear that the present strain is markedly related to Streptomyces 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, D., "International Journal of Systematic Bacteriology, 18, 68-189 and 279-392 (1968); 19, 391-512 (1969); 22, 265-394 (1972)"], the microorganism described in "The actinomycetes Vol. 2" written by Waksman [refer to Waksman, S.A., "The actinomycetes 2 (1961)"], with the microorganism described in Bergey's Manual edited by Buchanan and Gibbons [refer to R.E. Buchanan and N.E. Gibbons, "Bergey's Manual of Determinative Bacteriology", 8th edition (1974)], with the microorganism described in "Bergey's Manual of Systematic Bacteriology", edited by Williams [refer to Williams, S.T., et al., "Bergey's Manual of Systematic Bacteriology 4 (1989)"] and with the microorganism described in the recent literature about actinomycetes belonging to Streptomyces spp. It has however been recognized to be different from Streptomyces griseus, because it produces a yellowish grey soluble pigment on a glycerin -asparagine agar medium and a pale yellowish 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 water agar medium; the maximum growth temperature is 40°C; and it is grown in the presence of 7% 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 Streptomyces griseus SANK60196.

-58-
This strain was internationally deposited with Agency of Industrial Science and Technology, Ministry of Internet ional Trade and Industry (l-3,Higashi 1-chome, Tsukuba-shi, Ibaraki-ken, 305, JAPAN) as of February 22, 1996, with the accession number of FERM BP-5420.
A desertion was heretofo re made on Strain SANK60196. It Is known that various properties of actinomycetes are not fixed but easily change naturally or synthetically.
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-500359M-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 sate conventionally employed for cutevatem of the strain of the eumycetess or actinomycetes and are utilizable by a microorganism can be used as such nutrition sources
Specific examples of the carbon source Include glucose, fructose, maltose sucrose, marmttol, glycerol, dextrin, oats, rye, corn starch, potato, corn meat , soybean meal, cotton seed oil, thick malt syrup, therlac, soybean oil, citric acid and tartarlc acid. They may be used either singly or In combhation. The amount of the carbon source to be added usually varies, but not limited to, within a range of from 1 to 10 wt.%.
As the nitrogen source, a subsance containhg protein or hydrotyzate thereof can usually be employed. Deferred examples of the nitrogen source include soybean meal, wheat bran, peanut meat, cotton seed meal, casein hydroiyzate, Farrnamine, fish meal, com steep liquor, peptone, meat extract, pressed y east, dry yeast, yeast extract, matt
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 ranghg from 0.2 to 6 wt.% of the amount of the medium.

59
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-cysteine 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 mM permits favorable production of Compound A-500359M-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 8.0.
The temperature which allows Strain SANK60196 to grow ranges from 12 to 36°C. It is preferred to cultivate the strain at 18 to 28°C in order to produce Compound A-500359A, A-500359C, A-500359D, A-500359G or A-500359M-2, of which 19 to 23°C 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. Ordinarily-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 days at 19 to 23oC 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 Erlenmeyer 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

60
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 125°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, culturing is carried out with aeration and agitation at 19 to 23°C. This method is suitable for obtaining a large amount of compounds.
Compound A-500359M-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 during cultivation.
The production of Compound A-500359A, 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 litre of Compound A-500359A, 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, CHP-20P or HP-50, Sepabeads SP205, SP206 or SP207" (trade name; product of

61
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 of Nacalai Tesque); partition column chromatography using "Sephadex LH-20" (trade name; product of Pharmacia.Biotech); gel filtration chromato graphy using "Toyopearl HW40F" (trade name; product of TOSOH Corp); or high performance liquid chromatography using a normal phase or reversed phase column; or the like.
Compounds A-500359A, 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 activity can be determined by a method known to those skilled in the art.
Ester derivatives, ether derivatives and N-alkylcarbamoyl derivatives can each be prepared easily by using any one of the below-described Processes A to F or using them in combination as necessary.
(Process A)
Process A is for the preparation of an ester derivative of Compound (Ia) and by this process, Compound (Ic) wherein R2 is a methyl group can be prepared.

62
Process A

wherein: R1 and X have the same meanings as described above, R3b represents a hydrogen atom or a hydroxy-protecting group, R3b represents a hydrogen atom, a hydroxy-protecting group or an ester residue, R5b represents a hydrogen atom, a hydroxy-protecting group or an ester residue, R5b represents a hydrogen atom or a hydroxy-protecting group, and R3C represents a hydrogen atom, a hydroxy-protecting group or an ester residue, with the proviso that R3b and R5b do not represent a

63
hydrogen atom at the same time and R3C, R4b and Rsc do not all represent a hydrogen atom or a hydroxy-protecting group at the same time.
Step Al 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 (II).
Although the hydroxy-protecting step differs depending on the kind of the protecting group, it is conducted by a process well known in synthetic organic chemistry.
When the hydroxy -protecting group is a "silyl group", "alkoxymethyl group", "substituted ethyl group", "aralkyl group", "alkoxycarbonyl group", "alkenyloxycarbonyl group", "aralkyloxycarbonyl group", "1-(aliphatic acyloxy)-lower alkyl group", "1-(aliphatic acylthio)-lower alkyl group", "1-(cycloalkylcarbonyloxy)-lower alkyl group", "1-(aromatic acyloxy)-lower alkyl group", "1-(lower alkoxy carbon yloxy)-lower alkyl group", "1-(cyclo alkyl ox ycarbonyloxy)-lower alkyl group", "phthalidyl group", "oxodioxolenylmethyl group", "carbamoyl group substituted with 2 lower alkyl groups", "l-(lower alkoxycarbonyloxy)-lower alkyl group", "lower alkyl-dithioethyl group" or "1 -(acyloxy)-alkyloxycarbonyl group", this step is conducted by reacting Compound (II) with a desired hydroxy-protecting group halide in an inert solvent in the presence of a base.
Examples of the hydroxy-protecting group halide usable in the above reaction include trimethylsilyl chloride, triethylsilyl chloride, t-butyldimethylsilyl chloride, t-butyldimethylsilyl bromide, methyldi-t-butylsilyl chloride, methyldi-t-butylsilyl bromide, diphenylmethylsilyl chloride, diphenylmethylsilyl bromide, methoxymethyl chloride, 2-methoxyethoxymethyl chloride, 2,2,2-trichloroethoxymethyl chloride, 1-ethoxyethyl chloride, benzyl chloride, benzyl bromide, -naphthylmethyl chloride, diphenylmethyl chloride, diphenylmethyl bromide, triphenylmethyl chloride, 4-methylbenzyl chloride, 4-methoxybenzyl chloride, 4-nitrobenzyl chloride, 4-chlorobenzyl chloride, methoxycarbonyl chloride, ethoxycarbonyl chloride, 2,2,2-trichloroethoxycarbonyl chloride, vinyloxycarbonyl chloride, allyloxycarbonyl chloride, benzyloxycarbonyl chloride, benzyloxycarbonyl bromide, 4-methoxybenzyloxycarbonyl chloride, 4-nitrobenzyloxycarbonyl chloride, acetoxymethyl chloride, propionyloxymethyl chloride, butyryloxymethyl chloride, pivaloyloxymethyl chloride, pivaloyloxymethyl bromide, valeryloxymethyl chloride,

-64
1-acetoxyethyl chloride, butyryloxyethyl chloride, l-pivaloyloxyethyl chloride, cyclopentylcarbonyloxymethyl chloride, cyclohexylcarlbonyloxymethyl chloride, 1-cyclopentylcarbonyloxyethyl chloride, 1-cyclohexylcarbonyloxyethy! chloride, methoxycarbonyloxymethyl chloride, methoxycarbonyjoxymethyl bromide, ethoxycarbonyloxymethyl chloride, propoxycarbonyloxymethyl chloride, isopropoxycarbonyloxymethyl chloride, buioxycarbonyloxym ethyl chloride, isobutoxycarbonyloxymethyl chloride, l-(methoxycarbonyloxy)ethyl chloride, 1-(methoxycarbonyloxy)ethyl bromide, l-(ethoxycarbonyloxy)ethyl chloride, 1-(isopropoxycarbonyloxy)elhyl chloride, cyclopentyloxycarbonyloxymethyl chloride, cyclohexyloxycarbonyloxymethyl chloride, l-(cyclopentyloxycarbonyloxy)ethyl chloride, l-(cyclohexyloxycarbonyloxy)ethy! chloride, phthalidyl chloride, phthalidyl bromide, (5-phenyl-2-oxo-l,3-dioxolen-4-yl)methyl chloride, [5-(4-methy1phenyl)-2-oxo-l,3-dioxolen-4-yl]methyl chloride, (5-methyl-2-oxo-l,3-dioxolen-4-yl)methyl chloride, (5-methyl-2-oxo-l,3-dioxolen-4-yl)methyl bromide, (5-ethyl-2-oxo-l,3-dioxolen-4-yl)methyl chloride, dimethylcarbamoyl chloride, diethylcarbamoyl chloride, methyldithioethyl chloride, ethyldithioethyl chloride and pivaloyloxymethyloxycarbonyl chloride, of which triethylsilyl chloride, t-butyldimethylsilyl chloride, t-butyldimethylsilyl bromide, benzyl chloride, benzyl bromide, triphenylmethyl chloride, 4-methoxybenzyl chloride, 2,2,2-trichloroethoxycarbonyl chloride, allyloxycarbonyl chloride, benzyloxycarbonyl chloride, benzyloxycarbonyl bromide, acetoxymethyl chloride and pivaloyloxymethyl chloride are preferred.
Examples of the base include alkali metal hydroxides such as lithium hydroxide, 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, tributylamine, N-methylmorpholine, pyridine, 4-dimethylaminopyridine, picoline, lutidine, collidine, l,5-diazabicyclo[4.3.0]-5-noneneand l,8-diazabicyclo[5.4.0]-7-undecene. Out of these, organic amines are preferred, of which triethylamine, 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.

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 tetrafchloride 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-dimethylfonnamide, N,N-dirnethylacetamide, N-methyl-2-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 (IT), the halide and the solvent, it usually ranges from -10°C to 100°C (preferably 0 to 50°C). 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).
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, 3-bromodihydropyran, 4-methoxydihydropyran, dihydrothiopyran, 4-methoxydihydrothiopyran, 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 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.
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 1,2-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-pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethylsulfoxide; and mixtures thereof. Of these, hydrocarbons and ethers are preferred.

66
Although the reaction temperature differs with the nature of the starting compound (II), the cyclic ether compound and the solvent, it usually ranges from -10oC to 100°C (preferably 0 to 50°C). 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-prolecting group is a "carbamoyl group" or "carbamoyl group substituted with one lower alkyl group", Compound (II) is reacted with an isocyanate 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 above-exemplified 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-dim ethyl form amide, N,N-dimethyl acetamide, N-methyl-2-pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethylsulfoxide; and mixtures thereof. Of these, hydrocarbons and ethers are preferred.
Although the reaction temperature differs with the nature of the starting compound (II), the cyclic ether compound and the solvent, it usually ranges from -10°C to 100°C (preferably 0 to 50°C). 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

67
skilled in the art, for example, by recrystallization, column chromatography or the like.
Step A2 is for the preparation of a compound having the formula (Ic). 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 R6CO-Y, R6CO2CO2R9, R6CO-O-COR6 and R6OCO-Y [wherein R6 represents C6-20-alkyl, Y represents a halogen atom, preferably chlorine or bromine, R9 represents a C1-4 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 (R70)2PO-Y (wherein Y has the same meaning as described above and R represents a lower alkyl group), of which the compounds represented by any one of the formulas R6CO-Y, R6CO2CO2R9, R6CO-O-C0R6 and R6OCO-Y (wherein R6, Y and R9 have the same meanings as described above) are preferred.
Examples of the base usable in the above reaction include alkali metal hydroxides such as lithium hydroxide, 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, tributylamine, N-methylmorpholine, pyridine, 4-dimethylaminopyridine, picoline, lutidine, collidine, l,5-diazabicyclo[4.3.0]-5-nonene and l,8-diazabicyclo[5.4.0]-7-undecene. Of these, organic amines are preferred, of which triethylamine, 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

68
residue in an inert solvent in the presence of an acid or base, and oxidizing the reaction mixture into the corresponding phosphate estet by an oxidizing agent.
As the phosphite, a compound represented by the formula (R 7O)2-P-Z,. wherein R7 represents a C6-20 alkyl group and Z represents a'halogen atom or a compound represented by the formula -N(R8 )2 (wherein R represents a lower C6-20 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 meta-chloroperbenzoic acid, t-butylhydroperoxide and peracetic acid, of which meta-chloroperbenzoic 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-dim ethyl form amide, N,N-dimethylacetamide, N-methyl-2-pyrrolidone and hexamethylphosphoramide, and sulfoxides such as dimethyl sulfoxide; 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 -10oC to 100°C (preferably 0 to 50°C). 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 (III) 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 carbodiimides such as dicyclohexylcarbodiimide, carbonyl diimidazole and 1-(N,N-dimethylaminopropyl)-3-methylcarbodiimide hydrochloride, of which dicyclohexylcarbodiimide is 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,N-dimethylacetamide, 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 10 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, 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 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 "aralkyl group" or "aralkyloxycarbonyl group", deprotection is conducted by contacting 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 dioxane, aromatic hydrocarbons such as toluene,, benzine and xylene 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 above-exemplified 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, rhodium-aluminium 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 100°C (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, tetrahydrofuran and dioxane, amides such as dimethylformamide, dimethylacetamide and hexamethylphosphoramide 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 may be employed for oxidization. Examples include alkali metal persulfates such as potassium persulfate and sodium persulfate, cerie ammonium nitrate (CAN) and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), of which eerie 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°C to 150°C (preferably 0 to 50°C) and the reaction time usually ranges from 10 minutes to 24 hours (preferably 30 minutes to 10 hours).
When the hydroxy-protecting group is a t-butyl group, t-butoxycarbonyl group, "alkoxymethyl group", "tetrahydropyranyl or tetrahydrothiopyranyl group" or "tetrahydrofuranyl 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 50°C). Although the reaction time differs with the reaction temperature or the like, it ranges from 5 minutes to 48 hours (preferably 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 tetrabutylammonium 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, tetrahydrofuran 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°C (preferably

-72-
0 to 30°C) and the reaction time usually ranges from 2 to 24 hours (preferably 10 to 18 hours).
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.

73
(Process B)
Process B is for the preparation of an ester derivative of Compound (Ia). By this process, Compound (Id), wherein R2 is a methyl group, an -O- ester residue is present at the 2'-position, a hydroxy group or -O- ester residue is present at the 27-position and a hydroxy group or -O- ester residue is present at the 3"-position can be
Process B

wherein: R1 and X have the same meanings as described above, R3d represents an ester residue, R3b represents a hydrogen atom or an ester residue and R3d represents a hydrogen atom or an ester residue.

- 74
Step Bl is a step for preparing a compound of formula (IIIa). 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-dimeihoxypropane 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 p-toluenesulfonic acid, Lewis acids such as boron trifiuoride and acidic resins such as "Amberlyst 15", of which organic acids and acidic resins are preferred, with p-toluenesulfonic acid and "Amberlyst 15" being more preferred.
The reaction temperature usually ranges from -10 to 100°C (preferably 0 to 50°C). 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 by esterifying Compound (IIIa), removing an isopropylidene group from the esterified compound and then esterifying the hydroxy 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 Bl while using, as an inert solvent, water, an alcohol such as methanol or ethanol or aqueous alcohol.

75
(Process C)
Process C is for the preparation of an ester derivative of Compound (la). By this process, it is possible to prepare Compound (Ie) wherein R2 represents a methyl group, a protected or unprotected hydroxy group or an -O- ester residue is present at the 2"-poisition, and a protected or unprotected hydroxy group or an -O- ester residue is present at the 3"-position.
Process C

wherein: R1 and X have the same meanings as described above, R3e represents a hydrogen atom, a hydroxy-protecting group or an ester residue, and R5e represents a hydrogen atom, a hydroxy-protecting group or an ester residue, with the proviso that R3e and R5C 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 esterifying agent in an

76
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 (Ia). 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 -O- ester residue at the 2"-position and a protected or unprotected hydroxy group or an -O-ester residue at the 3"-position can be prepared.
Process D

wherein: R! and X have the same meanings as described above, R2a represents a hydrogen atom, a hydroxy-pro tec ting group or an ester residue, R3r represents a hydrogen atom, a hydroxy-protecting group or an ester residue, R4C represents a hydrogen atom, a hydroxy-protecting group or an ester residue, and R5f represents a

77-
hydrogen atom, a hydroxy-protecting group or an ester residue, with the proviso that all of R2a, R3f, R4C and R5f represent neither a hydrogenlatom nor a hydroxy-protecting group simultaneously.
Step Dl is a step for the preparation of Compound (If). 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 thehydroxy group if desired.
The protection of the diol portion with an isopropylidene group is conducted in a similar manner to that in Step Bl. Use of about 1 molar equivalent yields a mixture of a compound protected at the 2'- and 3'-positions and a compound protected at the 2"- and 3"-positions. The mixture can easily be separated, for example, by column chromatography.
Esterification is conducted in a similar manner to the corresponding reaction in Step A2. Use of an esterifying agent in an amount of about 1 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 1 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 hydroxy 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

78
mixture, for example, by column chromatography and if desired, protecting the hydroxy group. (Process E)
Process E is for the preparation of an ether derivative of formula (Ig) and (Ih) of Compound (Ia).

process E

79


wherein: R and X have the same meanings as described above, R represents the above-described ether residue and L represents a protecting group for the nitrogen atom of the uracil residue.

80



Step El is a step for preparing a compound represented by formula (TV) 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 acetoxyrnethyl chloride, of which 4-methoxybenzyloxymethyl chloride is preferred.
Examples of the base usable in the above reaction include tertiary amines such as l,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and l,5-diazabicyclo[4.3.0jnon-5-ene (DBN) and alkali metal hydrides such as sodium hydride and potassium hydride, of which l,8-diazabicyclo[5.4.0]undec-7-ene (DBU) is preferred.
Examples of the solvent usable in the above reaction include ethers such as diethyl ether, tetrahydrofuran and dioxane and amides such as N,N-dimethylformamide and N,N-dimethylacetamide, of which N,N-dimethylformamide is preferred.
The reaction temperature usually ranges from -30 to 100°C (preferably -10 to 30°C). 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 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 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, recrystallization, reprecipitation, column chromatography or the like.
Step E2 is a step for preparing a compound of the formula (V) 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 alkyl halides and alkyl triflates, of which an alkyl iodide is preferred.

Examples of the base usable in the above reaction include tertiary amines such as l,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and l,5-diazabicyclo[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 diethyl ether, tetrahydrofuran and dioxane and amides such as N,N-dimethylformamide and N.N-dimethylacetamide, of which N,N-dimethylformamide is preferred.
The reaction temperature usually ranges from -30 to 100°C (preferably -10 to 30°C). 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 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 E3 is a step for preparing a compound of the formula (Ig) by reacting a compound of the formula (V) with an agent capable of deprotecting the protected uracil residue in an inert solvent.
When the protecting group contained in the uracil residue in the formula (V) is a 4-methoxybenzyloxymethyl group, examples of the deprotecting agent usable here include 2,3-dichloro-5,6-dicyano-l,4-benzoquinone (DDQ) or cerium (TV) ammonium nitrate (CAN) [preferably 2,3-dichloro-5,6-dicyano-l,4-benzoquinone (DDQ)], 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 10 to 100°C). 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).

When the protecting group contained in the uracil group in the formula (V) 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 tetrahydrofuran, 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°C). Although the reaction time differs with the reaction temperature and the like, it usually ranges from 10 minutes to 1 day (preferably I 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, 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, column chromatography or the like.
Step E4 is a step for preparing a compound of the formula (Ih) by reacting a compound of the formula (Ig) with 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 "Amberlyst 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.

The reaction temperature usually ranges from 0 to 150°C (preferably 10 to 80°C). 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 hydroxy -protected compound, ester derivative or N-alkylcarbamoyl derivative by any one of Processes A to D and below-described Process F.

84
(Process F)
-Process F is for the preparation of an N-alkylcarbamoyl derivative of the invention compound (Ia). Process F

wherein: R1 and X have the same meanings as described above, R11 and R12 each independently represent the N-alkyl residue of the above-described N-alkylcarbamoyl group and Bz represents a benzoyl group.

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, l,8-diazabicyclo[5.4.0]undec-7-ene(DBU), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), pyridine and 4-dimethylaminopyridine and alkali metal hydrides 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 diethyl ether, tetrahydrofuran and dioxane, amides such as N,N-dimethyl form amide and N,N-dimethylacetamide, halogenated hydrocarbons such as methylene chloride and chloroform, and pyridine, of which pyridine is preferred.
The reaction temperature usually ranges from -30 to 100°C (preferably -10 to 30°C). 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, 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 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°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.

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-dimethylforrnamide and N,N-dimethylacetamide, of which alcohols are preferred.
The reaction temperature usually ranges from 0 to 100°C (preferably 10 to 60°C). 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, 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 chrom atography.
Compound (Ii) thus obtained can be converted to the corresponding hydroxy -protected compound, ester derivative or ether derivative by using any one of the above-described Processes A to E.

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The compounds of the present Invention or pharmacologically acceptable salts thereof may be administered through various routes. Examples include oral administration usng tablets, capsults, granules, powders, syrups or the Iike; 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 ana excipient, binder, disintegrator, lubricant, corrlgent, aojuvant for solubillzatlon, suspending agent, coating agent and/or the Ike.
For the formation of tablets, various carriers known conventionally In this field can be employed. Examples Include exclplents such as lactose, sucrose, sodium chloride, glucose,, urea, starch, calcium carbonate, Kaolh, crystaHhe cellulose and silicic acid; binders such as water, ethanol, propanol, simple syrup, glucose solutio n, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate and potyvinyl pyrrolidonhe; disinteojants such as dry starch, sodium aignate, agar powder, tamnaran 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 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 wt.%, preferably 1 to 30 wt.% in the whole composition.
There is no particular limitation on the administering method of the above-described 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


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 1 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.

90
[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 of Streptomyces 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 "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
CaCO3 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% (v/v) 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°C for 8 days. Main culture medium
Glucose 30 g
Meat extract 5 g
Polypeptone 5 g
Sodium chloride 5 g
CoCl2-6H2O 50 mg
CaCO3 3 mg
Antifoamer 50 mg

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("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 1) was filtered with the aid of "Celite 545" (product of Celite Co.) added at 4% (v/v). The filtrate (50 L) was charged on a "Diaion HP-20" column (product of Mitsubishi Chemical; 12 L). 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" 6 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 L). 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 A") 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 B") of 30% aqueous methanol eluate; and a peak at a retention time cf 22.6 minutes upon the above-described HPLC was detected from a 0 to 12 portion (which will hereinafter be called "Fraction C") of the 40% aqueous methanol eluate. These fractions were concentrated by "Evapor", respectively,


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 D") of the 40% 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 L). 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 40% aqueous methanol. From a combined fraction (which will hereinafter be called "Fraction E") 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 F") of the 1 to 3 L portion of the 30% 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 G") 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 H"), while Fraction C was combined with Fraction G (the combined one will hereinafter be called "Fraction I"). 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 of deionised water. The resulting solution was charged on a "Toyopearl HW-40F" column (product of TOSOH Corporation; 4 L), 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

(YMC-Pack ODS R-3105-20 (100 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 of A-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 of deionised water. The resulting solution was charged on a "Toyopearl HW-40F" column (8 L), 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 L). 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.
1) Appearance of the substance: white powder
2) Solubility: soluble in water and methanol, insoluble in normal hexane and
chloroform
3) Molecular formula: C14H33N5O12
4) Molecular weight; 583 (measured by FAB mass spectrometry)
5) Accurate mass, [M+H]+, as measured by high-resolution FAB mass spectrometry is
as follows:

Found: 584.2189 Calculated: 584.2205
6) Ultraviolet absorption spectrum: ultraviolet absorption spectrum measured in water
exhibits the following maximum absorption:
257 nm(e 10,300)
7) Optical rotation: optical rotation measured in methanol exhibits the following
value:
[ ]D20: +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, 1060 cm-1.
9) lH nuclear magnetic resonance spectrum was measured in deuterated methanol
with tetramethylsilane as an internal standard. 1H nuclear magnetic resonance
spectrum is as follows:
1.22(3H,d,J=6.7Hz), 1.29(lH,m), 1.49(lH,m), 1.78(lH,m), 1.87(lH,m), 1.92(lH,m),
2.01(lH,m), 3.44(3H,s), 3.58(lH,m), 3.86(lH,br.U=4.6Hz)t
3.96 (lH,ddd,J=0.7,4.5,5.7Hz), 4.30(lH,t,J=5.2Hz), 4.37(lH,t,J=4.lHz),
4.56(lH,dd,J=2.0,11.9Hz),4.58(lH,dd,J=2.0,4.3Hz),4.67(lH,d,J=2.0Hz),
5.23(lH,d,J=5.8Hz), 5.72(lH,d,J=8.1Hz), 5.88(IH,d,J=5.2Hz),
6.02(lH1br.dd,J=0.7,3.9Hz), 7.91(lH,d,J=8.lHz) ppm.
10) 13C nuclear magnetic resonance spectrum was measured in deuterated methanol
with tetramethylsilane as an internal standard. 13C nuclear magnetic resonance
spectrum is as follows:
22.2(q), 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), l09.3(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.

95
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 HW-40F" column (8.5 L), 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 140C18-OPN" column (product of Nacalai Tesque; 1.5 L). 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 10L 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 (100) 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: C23H31N5O12
4) Molecular weight: 569 (as measured by FAB mass spectrometry)
5) Accurate mass, [M+H]+, as measured by high-resolution FAB spectrometry is as
follows:

-96-
Found: 570.2034 Calculated: 570.2049
6) Ultraviolet absoiption 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:
[ ]D20: +89° (c 0.44, H2O)
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"1.
9) !H nuclear magnetic resonance spectrum was measured in deuterium oxide with the
signal of water as 4.75 ppm. 1H nuclear magnetic resonance spectrum is as follows:
1.20(3H,d,J=6.7Hz), 1.29(lH,m), 1.62(lH,m), 1.72(lH,m), 1.75(lH,m), 1.90(lH(m),
1.92(lH,m), 3.65(lH,m), 4.11(lH,dd,J=5.2,6.3Hz), 4.15(lH,ddd,J=1.4,4.2>4.3Hz),
4.18(lH,dd,J=3.3,5.2Hz), 4.43(lH,dd,J=2.1,6.3Hz), 4.49(lH,dd,J=3.0,4.4Hz),
4.62(lH,dd,J=1.7,l0.8Hz), 4.76(lH,d,J=2.lHz), 5.36(lH,d,J=4.0Hz),
5.77(lH,d,J=3.3Hz), S^tlHAJ^.lHz), 5.98(lH,br.dd,J=l.3,3.0Hz)J
7.72(lH,d,J=8.1Hz)ppm.
10) 13C nuclear magnetic resonance spectrum was measured in deuterium oxide with
1,4-dioxane (67.4 ppm) as an internal standard. I3C nuclear magnetic resonance
spectrum is as follows:
21.0(q), 26.8(t), 29.4(t), 35.4(t), 48.9(d), S2.6(d), 6l.9(d), 65.3(d), 69.4(d), 73.8(d), 76.7(d), 83.1(d), 89.7(d), lOO.l(d), 101.9(d), lO9.1(d), 141.0(d), 141.8(s), 151.6(s), 16L7(s), 166.4(s), 173.5(s), 175.8(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 Relent ion time: 13 minutes.

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 l portion of the resulting solution was charged on an HPLC column ("Senshu Pak Pegasil ODS " (20 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 35 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 35 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: C24H33N5O11
4) Molecular weight: 567 (as measured by FAB mass spectrometry)
5) Precise mass, [M+H]+, 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:
[ ]D20:+68°(c0.69,H2O)
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-1.

98
9) H nuclear magnetic resonance spectrum was measured in deuterium oxide with the
signal of water as 4.75 ppm. 1H [nuclear magnetic resonance spectrum is as follows:
1.12(3H,d,J=8.1Hz), 1.17(lH,m), 1.40(IH,m), 1.67(lH,m), 1.800H,m), 1.88(lH,m),
1.90(lH,m), 2.33(lH,m), 3.24(3H,s), 3.50(lH,m), 3.57(lH,t,J=4.7Hz),
4.08(lH,t,J=4.8Hz), 4.37(m),4.40(m), 4.46(lH,br.d,J=10.7Hz), 4.50(lH,d,J=2.0Hz),
5.30(IH,br.s), 5.64(lH,d,J=8.1Hz), 5.73(l-H,d,J=4.8Hz), 5.97(iH,d,J=2.4Hz),
7.77(lH,d,J=8.1Hz)ppm.
10) 13C nuclear magnetic resonance spectrum was measured in deuterated methanol
with the signal of methanol as 49.15 ppm. 13C 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(8), 175.5(s)ppm.
11) High performance liquid chromatography
Column: "Cosmosil 5C18-MS", 4.6 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 ran Retention time: 9.2 minutes.
Example 4: Cultivation of Streptomyces griseus Strain SANK 60196 (PERM 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°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
CaCO3 3 g

99

Cultivation was conducted as described below. Described specifically, the seed culture was inoculated at 3% (v/v) 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-(2-aminoethyl)-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.

Tap water 1000 ml
pH before sterilization: 7.4 Sterilization: at 121°C for 30 minutes

Example 5: Preparation of A-500359G (Exemp. compound No. 45)
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" 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: 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 L). 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 eiuted 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 1 portion of the resulting solution was charged on an HPLC column ("Senshu PakODS-H-5251" (20 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

101
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 1 portion of the resulting solution was charged on the above-described 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, whereby 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: C22H29N5O12
4) Molecular weight: 555 (as measured by FAB mass spectrometry)
5) Accurate mass, [M+H]+, 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:
[ ]D20:+109° (c 0.72, H2O)
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'1.
9) H nuclear magnetic resonance spectrum was measured in deuterium oxide with the
signal of water as 4.75 pprn. 1H nuclear magnetic resonance spectrum is as follows:

1.37 (lH.m), 1.65 (lH.m), 1.71 (1H, m), 1.79 (lH,m), 1.92 (1H, m), 1.98 (1H, m), 3.29 (1H, m), 3.36 (1H, m), 4.10 (1H, dd, J=5.0, 6.5 Hz), 4.14 (1H, dt, 1=3.5, 4.4 Hz), 4.17 (1H, dd,J=3.2, 5.0 Hz), 4.41 (1H, dd, J=2.1,' 6.5 Hz), 4.47 (1H, dd, J=2.9, 4.4 Hz), 4.61 (1H, dd, J=1.8, 11.4 Hz), 4.78 (1H), 5.35 (1H, d, J=4.1 Hz), 5.75 (1H, d, J=3.2 Hz), 5.82 (1H, d, J=8.2 Hz), 5.97 (1H, dd, J=1.5, 2.9 Hz), 7.71 (1H, d, J=8.2 Hz) ppm.
10) ' 3C nuclear magnetic resonance spectrum was measured in deuterium oxide with
1,4-dioxane (67.4 ppm) as an internal standard. 13C nuclear magnetic resonance
spectrum is as follows:
28.2 (t), 28.4 (t), 30.5 (t), 42.2 (t), 53.3 (d), 62.7 (d), 66.1 (d), 70.2 (d), 74.5 (d), 77.5 (d), 83.9 (d), 90.5 (d), 100.9 (d), 102.7 (d), 109.9 (d), 141.8 (d), 142.7 (s), 152.2 (s), 162.6 (s), 166.9 (s), 174.3 (s), 177.6 (s) ppm.
11) High performance liquid chromatography:
Column: "Senshu Pak ODS-H-2151", 6(J> 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 21 Onm Retention time: 4.6 minutes
Example 6: Cultivation of Streptomyces griseus Strain SANK60196 (FERM BP-5420)
Into each of four 2 L 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 S ANK60196, 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 g
Meat extract 5 g
Polypeptone * 5 g
Sodium chloride 5 g
CaCO3 3 g

103

The culture was conducted as described below. Described specifically, the seed culture broth was inoculated at 3% (v/v) 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-(2-aminoethyl)-L-cysteine hydrochloride was added to give a final concentration of 10 mM, and cultivation with aeration and agitation was then carried out for 6 days.

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.

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
30 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 L). 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

105
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: C23H31N5O12S
4) Molecular weight: 601 (as measured by FAB mass spectrometry)
5) Accurate mass, [M+H]+, 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( 14,000)
7) Optical rotation: optical rotation measured in water exhibits the following value: ?
[ ]D20:+58°(c0.39,H2O)
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
cm-1.
9) 'H nuclear magnetic resonance spectrum was measured in deuterium oxide with the
signal of water as 4.75 ppm. lH nuclear magnetic resonance spectrum is as follows:
1.30(3H,dJ=6.8Hz), 2.63(2H,m), 2.76(lH,dd,J=2.9,14.4Hz),
2.84(lH,dd,J=8.8,14.4Hz), 3.28(3H,s), 3.73(lH,dd,J=5.0,6.5Hz), 3.98(lH,m),
4.19(lH,ddd,J=1.5,3.5,4.4Hz), 4.38(IH,dd,J=3.2,5.0Hz), 4.47(1 H,dd,J=2.6,6.5Hz),
4.50(lH,dd,2.6,4.4Hz),4.73(lH,d,J=2.6Hz)>5.02(lH1dd,J=2.9,8.8Hz)5
5.39(m,dJ==3.5Hz), 5.75(lH,d,J=3.2Hz), 5.85(lH,d,J=8.1Hz),
6.03(lH,dd,J=1.5,2.6Hz),7.74(lH,dJ-8.1Hz)ppm.

106
,10)' C 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:
21.3(q), 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), 173.8(s), 174.5(s)ppm.
11) High performance liquid chromatography
Column: "Senshu Pak ODS-H-2i51", 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: 14.4 minutes
In the below-described Examples, Me, TBS, THF, TBAF, DMAP and WSC stand for a methyl group, a tert-butyldimethylsilyl group, tetrahydrofuran, tetrabutylammonium fluoride, 4-(dimethylamino)pyridine and l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride, respectively.

107
Example 8 (Exemp. compound No. 135)

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-butyldi methylsilyl 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 g), 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.

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:
5 - 7.99 (d, J = 8.1 Hz, 1H), 6.02 (d, J = 3.7 Hz, 1H), 5.88 (d, J = 5.1 Hz, 1H), 5.74 (d, J = 8.1 Hz, 1H), 5.23 (d, J = 5.8 Hz, 1H), 4.69 (s, 1H), 4.61 (d, J = 2.2 Hz, 1H), 4.51 (d, J = 11 Hz, 1H), 4.41 (t, J = 4.7 Hz, 1H), 4.36 (t, J = 4.6 Hz, 1H), 3.90 (m, 1H), 3.85 (m, 1H), 3.47 (s, 3H), 3.30-3.20 (m, 2H), 2.02 (m, 2H), 1.84 (m, 2H), L54-1.28 (m, 2H), 0.86 (s, 9H), 0.05 (s, 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-1.

108
(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 g), which was developed with methylene chloride - methanol (98:2 to 95:5), whereby 42.7 mg of the following compound was obtained.

1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilahe as an internal standard substance. 1H nuclear magnetic resonance spectrum is as follows:
8 = 9.17 (br s, 1H), 7.88 (m, 2H), 7.47 (br s, 1H), 6.58 (br s, 1H), 6.04 (m, 2H), 5.78 (m, 2H), 5.58 (m, 1H), 5.12 (d, J = 7.7 Hz, 1H), 4.64 (m, 1H), 4.60 (m, 1H), 4.50 (m, 2H), 4.06 (m, 1H), 3.88 (m, 1H), 3.46 (s, 3H), 3.27 (m, 3H), 2.37 (m, 2H), 2.16-1.10 (m, 32H), 0.88 (m, 12H), 0.06 (s, 6H) ppm. (8-3)
In 53 uL of THF were dissolved 41 mg of the compound obtained in (8-2). A 53 L THF solution containing 1M of TBAF 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 g), 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.

109

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:
8 -7.76 (d, J = 8.1 Hz, 1H), 5.88 (d, J = 3.7 Hz, lH), 5.79 (d, J = 5.1 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.42 (m, 1H), 5.21 (d, J = 4.7 Hz, 1H), 4.61 (d, J = 2.2 Hz, 1H), 4.54-4.46 (m, 2H), 4.17 (m, 2H), 3.71 (t, J = 4.8 Hz, 1H), 3.32 (s, 3H), 3.18 (m, 2H), 2.33 (t, J = 7.3 Hz, 2H), 1.98-0.79 (m, 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"1.

(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 ofeapuramycin was dissolved in 175 mL of acetone. To the resulting solution were added 9.2 mLof 2,2-dimethoxypropane and 253 mg of "Amberlyst 15 (H*)". The resulting mixture was stirred at room temperature. Two days later, the "Amberlyst 15 (H4)" 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

110
charged on a silica gel column (40 g), which was developed with methylene chloride -methanol (92:8), whereby 582.7 mg of the following compound was obtained.

1) 'H nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetram ethylsilane as an internal standard substance. 1H nuclear magnetic resonance spectrum is as follows:
6 = 9.69 (br s, 1H), 7.93 (d, J = 6.0 Hz, 1H), 7.74 (d, J = 8.2 Hz, 1H), 7.30 (br s, 1H), 7.03 (m, 1H), 6.34 (d, J " 4.4 Hz, 1H), 6.12 (br s, 1H), 5.92 (d, J = 6.4 Hz, 1H), 5.73 (d, J = 8.2 Hz, 1H), 4.82 (d, J = 7.2 Hz, 1H), 4.74 (m, 1H), 4.69 (m, 1H), 4.60 (m, 1H), 4.53 (m, 1H), 4.32 (m, 1H), 4.13 (t, J = 6.5 Hz, 1H), 4.02 (m, 1H), 3.69 (m, 1H), 3.50 (s, 3H), 3.28 (m, 2H), 2.18-1.70 (m, 6H), 1.49 (s, 3H), 1.45 (s, 3H) ppm.
(10-2)
In 3 mL of pyridine were dissolved 100 mg of the compound obtained in (10-1), 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.



(10-3)
In 10 mL of methanol was dissolved the whole amount of the mixture obtained in (10-2). To the resulting solution was added 100 mg of "Amberlyst 15 (H+)", and the mixture was stirred for 47 hours at room temperature and for 4 hours at 80°C. After filtration through Celite, the solvent was distilled off under reduced pressure. The residue was charged on a silica gel column (5 g), which was developed with methylene chloride - methanol (95:5 to 93:7), whereby 84.9 mg of the below-described compound was obtained as the desired compound of Example 10.

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 = 7.94 (d, J = 8.2 Hz, 1H), 6.01 (d, J = 3.5 Hz, 1H), 5.98 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.2 Hz, 1H), 5.42 (t, J = 4.8 Hz, 1H), 5.24 (d, J= 5.5 Hz, 1H), 4.68 (d, J= 1.8 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.05 (t, J = 4.8 Hz, 1H), 3.98 (t, J = 4.7 Hz, 1H), 3.38 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.01 (m, 2H), 1.84 (m, 2H), 1.63-1.15 (m, 28H), 0.90 (t, 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'1.
Example 14 (Exemp. compound No. 10)

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), 145 mg of

-112-
pent'adecanoic 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) 1H nuclear magnetic resonance spectrum was measured in deuterated methanol
with tetrametbylsilane as an internal standard substance. 'H nuclear magnetic
resonance spectrum is as follows:
S = 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.8 Hz, 1H), 5.97 (d, J = 4.9 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.44 (t, J = 4.8 Hz, 1H), 5.24 (d, J = 5.7 Hz, IK), 4,68 (d, J = 1.9 Hz, 1H), 4.55 (m, 2H), 4.42 (1, J = 4.1 Hz, 1H), 4.06 (t, J = 4.7 Hz, 1H), 3.97 (t, J = 5.0 Hz, 1H), 3.57 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.4 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.15 (m, 29H), 0.90 (t, 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'1.

The reaction was conducted in a similar manner to that described in Example 10 by using 100 mg of the compound obtained in Example (10-1) and 129 ^L of heptanoic anhydride, whereby 63.7 mg of the compound shown above was obtained as the desired compound of Example 15.
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:
5 = 7.94 (d, J = 8.2 Hz, 1H), 6.01 (d, J = 3.6 Hz, 1H), 5.97 (d, J = 4.9 Hz, 1H), 5.72 (d, J = 8.2 Hz, 1H), 5.42 (t, J - 4.9 Hz, 1H), 5.24 (d, J = 5.5 Hz, 1H), 4.68 (d, J = 2.0 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.2 Hz, 1H), 4.04 (t, J = 4.8 Hz, 1H), 3.98 (t, J =

113
4.9 Hz, 1H), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 (m, 2H), 1.83 (m, 2H), 1.63-1.25 (m, 10H), 0.9D (t, 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'1.

The reaction was conducted in a similar manner to that described in Example1 10 by using 100 mg of the compound obtained in Example (11-1), 158 mg of palmitic anhydride and 2 mg of DMAP, whereby 93.4 mg of the compound shown above was obtained as the desired compound of Example 16.
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 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.7 Hz, 1H), 5.98 (d, J = 4.9 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.6 Hz, 1H), 4.68 (d, J = 1.7 Hz, 1H), 4.55 (m, 2H), 4.41 (t, J = 4.2 Hz, 1H), 4.06 (t, J = 4.8 Hz, 1H), 3.97 (t, J = 4.7 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.3 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.20 (m, 31H), 0.90 (t, 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-1.

114



The reaction was conducted in a similar manner to that described in Example 10 by using 100 mg of the compound obtained in Example (11-1) and 177 |iLof 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 tetramethyJsilane as an internal standard substance. 1H nuclear magnetic
resonance spectrum is as follows:
6 - 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.8 Hz, 1H), 5.97 (d, J = 4.7 Hz, 1H), 5.72 (d, J - 8.1 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.4 Hz, 1H), 4.68 (d, J = 1.7 Hz, 1H), 4.55 (m, 2H), 4.41 (t, J = 4.1 Hz, 1H), 4.06 (t, J - 4.8 Hz, 1H), 3.97 (t, J = 5.0 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.4 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.20 (m, 19H), 0.90 (t, 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:
3390,2927,2855, 1689, 1510, 1459, 1430, 1384, 1336, 1269, 1151, 1109, 1062, 1022
cm"1.

The reaction was conducted in a similar manner to that described in Example 10 by using 100 mg of the compound obtained in Example (11-1) and 160 uL of pelargonic anhydride, whereby 59.9 mg of the desired compound shown above was obtained.

- 115
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 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.8 Hz, 1H), 5.97 (d, J = 4.7 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.6 Hz, 1H), 4.68 (d, J= 1.6 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.06 (t, J = 4.8 Hz, 1H), 3.97 (t, J = 4.9 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.3 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.20 (m, 17H), 0.90 (t, J = 6.6 Hz, 3H) pprn.
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-1.
Example 19 (Exemp. compound No. 9)

The reaction was conducted in a similar manner to that described in Example 10 by using 100 mg of the compound obtained in Example (11-1) and 105 mgof myristic anhydride, whereby 81.6 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 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.6 Hz, 1H), 4.68 (d, J = 1.8 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.06 (t, J = 4.8 Hz, 1H), 3.97 (t, J = 4.9 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.3 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.20 (m, 27H), 0.90 (t, 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 cm1.

116



The reaction was conducted in a similar manner to that described in Example 10 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) 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:
= 7.95 (d, J = 8.2 Hz, 1H), 6.01 (d, J = 3,9 Hz, 1H), 5.97 (d, J = A.I Hz, 1H), 5.72 (d, J = 8.2 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.7 Hz, 1H), 4.69 (d, J = 1.6 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.07 (t, J = 4.8 Hz, 1H), 3.97 (t, J = 4.7 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.37 (t, J = 7.3 Hz, 2H), 2.05-1.75 (m, 4H), 1.63-1.20 (m, 23H), 0.90 (t, 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"1.

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 of oleic acid, whereby 70.9 mg of the compound shown above was obtained.

-117-
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:
5 = 7.95 (d, J = 8.2 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.2 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.34 (t, J - 4.8 Hz, 2H), 5.24 (d, J = 5.7 Hz, 1H), 4.68 (d, J = 1.9 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4 J Hz, 1H), 4.07 (t, J = 4.8 Hz, 1H), 3.97 (t, J = 4.7 Hz, 1H), 3.58 (m, 1H), 3.38 (s; 3H), 2.37 (t, J = 7.4 Hz, 2H), 2.05-1.75 (m, 8H), 1.60 (m, 2H), 1.49 (m, 1H), 1.33 (m, 21H), 1.22 (d, J = 6.7 Hz, 3H), 0.89 (t, 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* .

The reaction was conducted in a similar manner to that described in Example 10 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 tetrarnethylsilane as an internal standard substance. H nuclear magnetic resonance spectrum is as follows:
6 - 7.95 (d, J = 8.0 Hz, 1H), 6:01 (d, J = 3.8 Hz, 1H), 5.97 (d, J - 4.8 Hz, 1H), 5.72 (d, J = 8.0 Hz, 1H), 5.45 (t, J = 4.9 Hz, 1H), 5.34 (m, 6H), 5.24 (d, J = 5.7 Hz, IH), 4.68 (d, J = 1.9 Hz, 1H), 4.55 (m, 2H), 4.41 (t, J = 4.2 Hz, 1H), 4.07 (t, J = 4.8 Hz, 1H), 3.97 (t, J " 4.8 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.81 (t, J = 5.9 Hz, 4H), 2.38 (t, J " 7.3 Hz, 2H), 2.10-1.75 (m, 8H), 1.60 (m, 2H), 1.49 (m, 1H), 1.32 (m, 9H), 1.22 (d, J = 6.7 Hz, 3H), 0.97 (t, J = 7.5 Hz, 3H) ppm.

- 118-
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'1.

The reaction was conducted in a similar manner to that described in Example 10 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:
5 - 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.45 (t, J - 4.9 Hz, 1H), 5.35 (m, 4H), 5.24 (d, J = 5.7 Hz, 1H), 4.68 (d, J " 1.9 Hz, 1H), 4.55 (m, 2H), 4.41 (t, J = 4.2 Hz, 1H), 4.07 (t, J = 4.8 Hz, 1H), 3.97 (t, J * 5.0 Hz, 1H), 3.58 (m, 1H), 3.38 (s, 3H), 2.77 (t, J = 6.3 Hz, 2H), 2.38 (t, J = 7.3 Hz, 2H), 2.10*1.75 (m, 8H), 1.60 (m, 2H), 1.49 (m, IH), 1.32 (m, 15H), 1.22 (d, J = 6.7 Hz, 3H), 0.97 (t, 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-1.

- 119-

The reaction was conducted in a similar manner to that described in Example 10 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) lH nuclear magnetic resonance spectrum was measured in deuterated methano]
with tetramethylsilane as an internal standard substance. 1H nuclear magnetic
resonance spectrum is as follows:
5 = 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.42 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.7 Hz, 1H), 4.68 (d, J = 1.6 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.04 (t, J = 4.8 Hz, 1H), 3.98 (t, J = 4.8 Hz, 1H), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 (m, 2H), 1.84 (m, 2H), 1.64-1.25 (m, 20H), 0.90 (t, 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'1.

The reaction was conducted in a similar manner to that described in Example 10 by using 150 mg of the compound obtained in Example (10-1) and 181 ul of decanoic anhydride, whereby 124.3 mg of the compound shown above 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:
8 - 7.94 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.9 Hz, 1H), 5.72 (d, J - 8.1 Hz, 1H), 5.42 (t, J = 4.8 Hz, 1H), 5.24 (d, J = 5.6 Hz, 1H), 4.68 (d, J = 1.7 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.2 Hz, 1H), 4.04 (t, J = 4.8 Hz, 1H), 3.98 (t, J = 4.8 Hz, 1H), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 (m, 2H), 1.84 (m, 2H), 1.64-1.25 (m, 16H), 0.90 (t, 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)

The reaction was conducted in a similar manner to that described in Example 10 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:
5 = 7.94 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.42 (t, J - 5.0 Hz, 1H), 5.24 (d, J - 5.6 Hz, 1H), 4.68 (d, J = 1.6 Hz, 1H), 4.55 (m, 2H), 4,42 (t, J = 4.1 Hz, 1H), 4.04 (t, J "= 4.8 Hz, 1H), 3.98 (t, J = 4.9 Hz, 1H), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 (m, 2H), 1.84 (m, 2H), 1.64-1.25 (m, 24H), 0.90 (t, 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'.


- 121
Example 27 (Exemp. compoundiNo. 48)
The reaction was conducted in a similar manner to that described in Example 10 by using 150 mg of the compound obtained.in Example (10-1) and 163 |il 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:
5 = 7.94 (d, J = 8.1 Hz, 1H), 6.01 (d, J - 3.8 Hz, 1H), 5.97 (d, J = 5.0 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.42 (t, J - 4.8 Hz, 1H), 5.24 (d, J = 5.4 Hz, 1H), 4.68 (d, J * 1.8 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.2 Hz, 1H), 4.04 (t, J = 4.8 Hz, 1H), 3.98 (t, J = 4.9 Hz, 1H), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 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"1.


122
Example 29 (Exemp. compound No. .52)
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.
1) !H nuclear magnetic resonance spectrum was measured in deuterated methanol
with tetrarnethylsilane as an internal standard substance. lH nuclear magnetic
resonance spectrum is as follows:
6 = 7.94(d,J = 8.1 Hz, 1H), 6.01 (d,J = 3.7Hz, 1H), 5.97 (d, J = 5.0 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.42 (t, J = 4.9 Hz, 1H), 5.24 (d, J = 5.6 Hz, 1H), 4.68 (d, J = 2.0 Hz, 1H), 4.55 (m, 2H), 4.42 (t, J = 4.1 Hz, IH), 4.04 (t, J = 4.8 Hz, IH), 3.98 (t, J = 4.8 Hz, IH), 3.37 (s, 3H), 3.25 (m, 2H), 2.37 (t, J = 7.3 Hz, 2H), 2.00 (m, 2H), 1.84 (m, 2H), 1.64-1.25 (m, 26H), 0.90 (t, 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'1.
Example 31 (Exemp. compound No. 5)

The reaction was conducted in a similar manner to that described in Example 10 by using 187 mg of the compound obtained in Example (11-1) and 267 [il of

123
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:
6 = 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.97 (d, J = 4.9 Hz, 1H), 5.72 (d, J- 8.1 Hz, 1H), 5.44 (t, J = 4.9 Hz, 1H), 5.23 (d, J = 5.5 Hz, 1H), 4.68 (d, J = 2.0 Hz, 1H), 4.56 (m, 1H), 4.52 (m, 1H), 4.42 (t, J = 4.1 Hz, 1H), 4.06 (t, J = 4.7 Hz, 1H), 3.97 (t, J = 5.1 Hz, 1H), 3.57 (m, 1H), 3.38 (s, 3H), 2.37 (U J = 7.3 Hz, 2H), 2.05-1.75 (m, 4H), 1.60 (m, 2H), 1.48 (m, 1H), 1.32 (m, 9H), 1.21 (d, J = 6.6 Hz, 3H), 0.90 (t, 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-1.

In 3 mL of pyridine were dissolved 125 mg of the compound obtained in Example (11-1), 170 ul of nonyl chloro formate, 147 mg of dim ethyl aminopyridine 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 NaHCO3 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

124
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. 1H nuclear magnetic
resonance spectrum is as follows:
5 = 7.94 (d, J = 8.2 Hz, 1H), 6.01 (d, J = 4.0 Hz, IH), 5.98 (d, J =4.6 Hz, 1H), 5.71 (d, J = 8.2 Hz, 1H), 5.32 (t, J = 4.8 Hz, 1H), 5.23 (d, J = 5.7 Hz, IH), 4.68 (d, J = 2.0 Hz, 1H), 4.56 (m, IH), 4.52 (m, IH), 4.41 (t, J = 4.2 Hz, IH), 4.13 {m, 3H), 3.97 (t, J - 5.0 Hz, 1H), 3.57 (m, IH), 3.40 (s, 3H), 2.05-1.75 (m, 4H), 1.65 (m, 2H), 1.48 (m, 1H), 1.32 (m, 13H), 1.22 (d, 1 = 6.6 Hz, 3H), 0.90 (t, 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
cm"'.
Example 33 (Exemp. compound No, 539)

The reaction was conducted in a similar manner to that described in Example 32 except for the use of 157 ul 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 methano] with tetramethylsilane as an internal standard substance. *H nuclear magnetic resonance spectrum is as follows:
8 = 7.94 (d, J = 8.1 Hz, IH), 6.01 (d, J = 3.9 Hz, IH), 5.98 (d, J = 4.4 Hz, IH), 5.71 (d, J = 8.1 Hz, IH), 5.32 (t, J = 4.6 Hz, IH), 5.24 (d, J = 5.6 Hz, IH), 4.69 (d, J - 2.0 Hz, IH), 4.56 (m, IH), 4.52 (m, IH), 4.41 (t, J = 4.0 Hz, IH), 4.13 (m, 3H), 3.97 (t, J

125
= 5:0 Hz, 1H), 3.57 (m, 1H), 3.40 (s, 3H), 2.05-1.75 (m, 4H), 1.65 (m, 2H), 1.48 (m, 1H), 1.32 (m, 11H), t.22 (d, J * 6.6 Hz, 3H), 0.90 (t, 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"1.

126
Example 34 (Exemp. compound No. 594)

In 50 mL of dimethylformamide (DMF) were dissolved 4.57 g of the compound obtained in Example (11-1) and 2.2 mL of l,8-diazabicyclo[5.4.0]-7-undecene (DBU). To the resulting solution was added a solution obtained by dissolving 2.45 g of 4-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 anhydrous 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% methanol in melhylene chloride, whereby 4.80 g of the desired compound was obtained.
1) lH nuclear magnetic resonance spectrum was measured in deuterated chloroform with tetramethylsilane as an internal standard substance. ]H nuclear magnetic resonance spectrum is as follows:
5 - 7.85 (m, 1H), 7.69 (d, J = 8.2 Hz, 1H), 7.32 (m, 2H), 7.15 (m, 2H), 6.85 (d, J = 8.7 Hz, 2H), 6.37 (d, J = 4.3 Hz, 1H), 6.06 (d, J - 6.2 Hz, 1H), 5.82 (m, 1H), 5.75 (d, J = 8.2 Hz, 1H), 5.70 (m, 1H), 5.44 (m, 2H), 4.73 (m, 3H), 4.61 (s, 2H), 4.57 (s, 1H), 4.45 (m, 1H), 4.25 (m, 1H), 4.03 (m, 2H), 3.79 (s, 3H), 3.56 (s, 3H), 3.53 (mt 1H), 3.28 (d,

-127-
J = 7.8 Hz, 1H), 2.35 (s, 2H), 2.15 (rn, 1H), 2.02-1.75 (m, 4H), 1.49 (s, 3H), 1.42 (s, 3H), 1.30 (m, 2H), 1.23 (d, 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"1.

In 5 mL of DMF was dissolved 773 mg of the compound obtained in Example (34-1). The resulting solution was stirred at0°C under a nitrogen gas stream. To the reaction mixture was added 60 mg of NaH (about 60%). 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. 'H nuclear magnetic resonance spectrum is as follows:
5 = 7.89 (d, J = 8.1 Hz, 1H), 7.75 (d, J = 5.9 Hz, 1H), 7.31 (d, J = 8.8 Hz, 2H), 7.13 (br s, 1H), 6.86 (d, J = 8.8 Hz, 2H), 6.37 (m, 1H), 5.95 (s, 1H), 5.75 (br s, 1H), 5.70 (d, J = 8.1 Hz, 1H), 5.57 (m, 1H), 5.45 (s, 2H), 4.78 (d, J - 8.1 Hz, 1H), 4.74 (m, 2H),

128
4.63 (s, 2H), 4.55 (s, 1H), 4.46 (m, 1H), 4.05 (m, 2H), 3.95 (m, 1H), 3.79 (s, 3H), 3.62 (m, JH), 3.51 (m, 1H), 3.43 (s, 3H), 4.09 (m, 1H)T 1.98 tm, 1H), 1.86 (m, 1H), 1.77 (m, 1H), 1.49 (s, 3H), 1.44 (s, 3H), 1.40-1.20 (m, 18H), 1.19 (d, J = 6.6 Hz, 3H), 0.88 (t, 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, 3102, 2928, 2855,1713, 1670, 1613, 1587, 1514, 1456, 1382, 1359, 1338, 1300, 1271,1248,1220,1167, 1112, 1066, 1013 cm"1.
(34-3)

In 5 mL of methylene chloride was dissolved 390 mg of the compound obtained in Example (34-2). To the resulting solution were added 276 L of water and 484 mg of 2,3-dichloro-5,6-dicyano-l,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 tetramethylsilane as an internal standard substance. 'H nuclear magnetic resonance spectrum is as follows:
6 = 9.30 (br s, 1H), 7.99 (d, J= 7.3 Hz, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.19 (br s, 1H), 6.36 (d, J = 4.4 Hz, 1H), 5.98 (br s, 1H), 5.85 (br s, 1H), 5.81 (d, J = 5.1 Hz, 1H), 5.69 (dd, J =2.2 and 8.1 Hz, 1H), 4.74 (m, 2H), 4.60 (m, 2H), 4.28 (t, J = 4.7 Hz, 1H), 4.12 (t, J= 6.2 Hz, 1H),4.07 (t, J =4.7 Hz, 1H), 3.59 (m, 3H), 4.43 (s, 3H), 2.10-1.73 (m,

4H), 1.60 (m, 2H), 1.48 (s, 3H), 1.42 (s, 3H), 1.23 (m, 19H), 0.88 (t, 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.

In 15 mL of methanol was dissolved 273 mg of the compound obtained in Example (34-3). To the resulting solution was added 260 mg of "AmberJyst ] 5" 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 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 tetramethylsilane as an internal standard substance. *H nuclear magnetic
resonance spectrum is as follows:
8 = 7.95 (d, J = 8.1 Hz, 1H), 6.02 (d, J = 3.6 Hz, 1H), 5.92 (d, J = 4.5 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.23 (d, J = 5.3 Hz, 1H), 4.67 (s, 1H), 4.59 (m, 1H), 4.52 (m, 1H), 4.38 (t, J = 4.2 Hz, 1H), 4.08 (t, J = 4.6 Hz, 1H), 3.98 (t, J = 4.7 Hz, 1H), 3.94 (t, J = 4.7 Hz, 1H), 3.58 (m, 3H), 3.40 (sf 3H), 2.05-1.75 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"1.

130
Example 35 (Exemp. compound No. 590) (35-1)

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) '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:
5 = 7.91 (d, J - 8.3 Hz, 1H), 7.24 (d, J - 8.6 Hz, 2H), 6.85 (d, J = 8.6 Hz, 2H), 6.18 (d, J = 4.1 Hz, 1H), 5.92 (d, J = 4.0 Hz, IH), 5.74 (d, J = 8.3 Hz, 1H), 5.42 (s, 2H), 5.11 (d, J = 5.4 Hz, 1H), 4.80 (m, 1H), 4.70 (m, 1H), 4.55 (m, 3H), 4.37 (t, J = 5.8 Hz, 1H), 4.08 (t, J = 4.3 Hz, 1H), 3.94 (t, J = 5.2 Hz, 1H), 3.76 (s, 3H), 3.60 (m, 3H), 3.41 (s, 3H), 2.05-1.75 (m, 4H), 1.55 (m, 3H), 1.43 (s, 6H), 1.25 (m, 8H), 1.19 (d, J = 6.6 Hz, 3H), 0.88 (t, 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 cm1.

-131
(35-2)

The reaction was conducted in a similar manner to that described in Example (34-3) using 458 mg of the compound obtained in Example (35-1), 313 mg of the desired compound was obtained.
1) H nuclear magnetic resonance spectrum was measured in deuterated chloroform
with terramethylsilane as an internal standard substance. H nuclear magnetic
resonance spectrum is as follows:
8 = 9.28(brs,lH), 7.99 (d,J = 6.6 Hz, 1H), 7.71 (d,J = 8.1 Hz, 1H),7.19 (br s, 1H), 6.36 (d, J = 4.4 Hz, lH),5.98(brs, 1H), 5.85 (brs, 1H), 5.81 (d, J = 5.1 Hz, 1H), 5.69 (dd, J = 2.2 and 8.1 Hz, 1H), 4.74 (m, 2H), 4.60 (m, 3H), 4.28 (t, J = 4.7 Hz, 1H), 4.12 (t, J = 6.9 Hz, 1H), 4.07 (t, J = 4.7 Hz, 1H), 3.59 (m, 3H), 4.42 (s, 3H), 2.10-1.73 (m, 4H), 1.60 (m, 2H), 1.48 (s, 3H), 1.42 (s, 3H), 1.23 (m, 11H), 0.87 (t, 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 cm-1.
(35-3)
In 15 mL of methanol was dissolved 273 mg of the compound obtained in Example (35-2). To the resulting solution was added 260 mg of "Amberlyst 15". The resulting mixture was stirred at 80°C. After 4 hours and 20 minutes, the insoluble

-132-
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 tetramethylsilane as an internal standard substance. !H nuclear magnetic
resonance spectrum is as follows:
6 = 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 3.9 Hz, 1H), 5.92 (d, J = 4.5 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.23 (d, J = 5.6 Hz, 1H), 4.66 (d, J = 2.0 Hz, 1H), 4.59 (m, 1H), 4.50 (m, 1H), 4.38 (t, J = 3.9 Hz, 1H), 4.08 (t, J = 4.7 Hz| 1H), 3.99 (t, J = 4.9 Hz, 1H), 3.93 (t, J = 4.7 Hz, 1H), 3.58 (m, 3H), 3.40 (s, 3H), 2.05-1.75 (m, 4H), 1.52 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'1.
Example 36 (Exemp. compound No. 891)

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

133
with 200 mL each of saturated aqueous sodium bicarbonate and saturated and then
dried over anhydrous sodium sulfate. The solvent wasidistilled off under reduced
pressure and the residue was charged on a silica gel column (50 g), 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 (m, 4H), 7.92 (m, 4H), 7.55 (m, 5H), 7.40 (m, 5H), 7.15 (br
s, 1H), 6.45 (br s, 1H), 6.32 (d, J = 3.7 Hz, 1H), 6.13 (m, 1H), 6.09 (br s, 1H), 5.96 (d,
J = 3.7 Hz, 1H), 5.83 (m, 2H), 5.62 (m, 2H), 4.69 (m, 1H), 4.61 (m, 1H), 4.56 (m,
1H), 4.36 (t, J = 5.9 Hz, 1H), 3.54 (m, 1H), 3.34 (s, 3H), 2.12 (m, 1H), 2.00-1.50 (m,
4H), 1.32 (m, IH), 1.24 (d, J = 6.6 Hz, 3H) ppm.
(36-2)

In 6.3 mL of methylene chloride was dissolved 418 mg of the compound obtained in Example (36-1). To the resulting solution was added 5 mL of water, followed by stirring at room temperature. To the reaction mixture, 4.74 g of nitrosylsulfuric acid was gradually added over 30 minutes. After stirring for a further 10 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-N-nitrosourea, 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 (20 g) which was developed with 1.5% methanol in methylene chloride, whereby 99 mg of the desired compound was obtained.

1) 1H nuclear magnetic resonance spectru'm was measured in deuterated chloroform
with letramethylsilane as an internal standard substance. 'H nuclear magnetic
resonance spectrum is as follows;
8 = 8.28 (s, 1H), 8.06 (d, J = 7.3 Hz, 2H), 7.99 (d, J = 7.3 Hz, 2H), 7.95 (m, 3H), 7.60-7.32 (m, 11H), 6.33 (s, IH), 6.20 (t, J = 3.6 Hz, 1H), 6.06 (d, J =4.4 Hz, 1H), 5.94 (d, J = 5.9 Hz, 1H), 5.88 (t, J = 4.0 Hz, IH), 5.70 (d, J = 3.7 Hz, 1H), 5.54 (m, 2H), 4.79 (m, 1H), 4.63 (m, IH), 4.17 (t, J = 5.5 Hz, 1H), 3.83 (s, 3H), 3.80 (m, 1H), 3.72 (m, 1H), 3.35 (m, IH), 3.30 (s, 3H), 2.19 (m, 1H), 2.02-1.75 (m, 3H), 1.52 (m, 1H), 1.32 (m, 1H), 1.24 (d, 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'1.
(36-3)
In 2 mL of a 40% methylamine - methanol solution was dissolved 98 mg of the compound obtained in Example (36-2). 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 30 mg of the desired compound was obtained.
1) *H nuclear magnetic resonance spectrum was measured in deuterated melhanol with tetramethylsilane as an internal standard substance. JH nuclear magnetic resonance spectrum is as follows:
5 = 7.86 (d, J = 8.0 Hz, IH), 5.98 (m, IH), 5.83 (m, IH), 5.74 (dd, J = 2.9 and 8.1 Hz, IH), 5.24 {d, J = 4.9 Hz, IH), 4.73 (dd, J = 2.1 and 10.9 Hz, IH), 4.50 (m, 2H), 4.38 (t, J - 4.0 Hz} IH), 4.25 (m, IH), 4.04 (m, 2H), 3.75 (m, IH), 3.39 (d, J = 2.8 Hzt 3H), 2.74 (d, J = 2.4 Hz, 3H), 1.65 (m, IH), 1.25 (m, 2H), 1.00 (m, 3H), 0.92 (m, 1H)( 0.75 (m, 2H) ppm.

-135
Example 37 (Exemp. compound No. 991)

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 (36-2), 0.4 mL of n-propylamine 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 tetram ethyl si lane as an internal standard substance. 'H nuclear magnetic
resonance spectrum is as follows:
8 = 7.91 (d, J = 8.1 Hz, 1H), 6.02 (d, J = 4.2 Hz, 1H), 5.89 (d, J = 5.5 Hz, 1H), 5.72 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, 2934,2878, 1683,1515,1459,1432, 1385,1335, 1269,1140,
1080,1062, 1022,981 cm"1.

136
Example 38 (Exemp. compound No. 1091)

The reaction was conducted in a similar manner to that described in Example (36-3) using 270 mg of the compound obtained'in Example (36-2), 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. ]H nuclear magnetic
resonance spectrum is as follows:
8 = 7.92(d,J = 8.1 Hz, 1H), 6.02 (d, J-4.4 Hz, 1H), 5.91 (d, J = 5.9 Hz, 1H), 5.73 (d, J = 8.1 Hz, 1H), 5.15 (d, J = 5.9 Hz, 1H), 4.67 (d, J = 2.2 Hz, 1H), 4.55 (m, 2H), 4.36 (t, J = 4.4 Hz, 1H), 4.32 (t, J = 5.5 Hz, 1H), 3.92 (m, 2H), 3.60 (m, 1H), 3.47 (s, 3H), 3.35 (m, 1H), 3.20 (m, 1H), 2.05-1.75 (m, 4H), 1.50 (m, 3H), 1.28 (m, 19H), 1.22 (d, J = 6.6 Hz, 3H), 0.89 (t, 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"1.
Example 39 (Exemp. compound No. 548)



137
(39-1)
In 4 mL of pyridine was dissolved 125 mg of the compound obtained in Example (11-1). Under a nitrogen gas stream, 147 mg of dimethytaminopyridine and 3.9 mg of 4-pyrroljdinopyridine were added to the solution. After cooling to 0°C, 209.1 mgof 2,2-dimethyldodecanoyl chloride (B.D. 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 0cC, 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 anhydrous 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:
8 = 7.90 (d, J = 8.1 Hz, 1H), 6.16 (d, J = 3.7 Hz, 1H), 6.03 (d, J = 5.4 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.32 (t, J = 5.2 Hz, 1H), 5.14 (d, J = 5.3 Hz, 1H), 4.90 (m, 1H), 4.75 (d, J = 2.1 Hz, 1H), 4.59-4.55 (m, 2H), 4.38 (t, J = 5.8 Hz, 1H), 4.05 (t, J = 4.4 Hz, 1H), 3.64-3.55 (m, 1H), 3.40 (s, 3H), 2.01-1.77 (m, 4H), 1.59-1.47 (m, 3H), 1.45 (s, 6H), 1.34-1.10 (m, 26H), 0.89 (t, J = 6.7 Hz, 3H) ppm.


138
(39-2)
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. lH nuclear magnetic
resonance spectrum is as follows:
5 = 7.95 (d, J = 8.1 Hz, 1H), 6.02 (d, J = 3.9Hz, 1H), 5.98 (d, J = 4.8 Hz, 1H), 5.71 (d, J = 8.1 Hz, 1H), 5.39 (t, J = 4.8 Hz, 1H), 5.24 (d, J = 5.4 Hz, 1H), 4.69 (d, J = 2.1 Hz, IH), 4.57-4.56 (m, lH), 4.54-4.50 (m, 1H), 4.42 (t, J = 4.1 Hz, 1H), 4.06 (t, J = 4.8 Hz, 1H), 3.98 (t, J = 4.9 Hz, 1H), 3.61-3.53 (m, IH), 3.37 (s, 3H), 2.04-1.76 (m, 4H), 1.56-1.43 (m, 2H), 1.33-1-.16 (m, 27H), 0.89 (t, 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"'.

139
Example 40 (Exemp. compound No. 574) OH



(40-1)
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 (11-1), 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 tetramethylsilane as an internal standard substance. 'H nuclear magnetic
resonance spectrum is as follows:
5 = 7.90 (d, J = 8.1 Hz, 1H), 6.16 (d, J = 3;7 Hz, IH), 6.03 (d, J * 5.7 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.30 (t, J = 5.3 Hz, 1H), 5.15 (d, J = 5.4 Hz, IH), 4.90 (m, IH), 4.75 (d, J = 2.1 Hz, IH), 4.59-4.57 (m, 2H), 4.39 (t, J = 5.9 Hz, IH), 4.03 (t, J = 4.4 Hz, IH), 3.39 (s, 3H), 3.31-3.28 (m, 2H), 2.02 (d, J = 11 Hz, 2H), 1.87-1.77 (m, 2H), 1.60-1.49 (m, 2H), 1.44 (s, 6H), 1.40-1.20 (m, 18H), 1.17 (s, 6H), 0.89 (t, 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"1.
(40-2)

140

In a similar manner to that described in Example (39-2) except for the use of 95.3 nag of the compound obtained in Example (40-1) instead of the compound obtained in Example (39-1), 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 tetramethylsilane as an internal standard substance. H nuclear magnetic
resonance spectrum is as follows:
6 = 7.95 (d, J = 8.2 Hz, 1H), 6.02 (d, J = 3.8 Hz, 1H), 5.98 (d, J = 4.8 Hz, 1H), 5.72 (d, J = 8.2 Hz, 1H), 5.37 (t, J = 5.0 Hz, 1H), 5.24 (d, J = 5.4 Hz, 1H), 4.68 (d, J = 2.1 Hz, 1H), 4.57-4.52 (m, 2H), 4.42 (t> J - 4.1 Hz, 1H), 4.04 (t, J = 4.9 Hz, 1H), 3.98 (t, J = 4.8 Hz, 1H), 3.37 (s, 3H), 3.27-3.22 (m, 2H)S 2.04-1.89 (m, 2H), 1.86-1.77 (m, 2H), 1.58-1.46 (m, 2H), 1.43-1.19 (m, 18H), 1.16 (d, J = 6.2 Hz, 6H), 0.89 (t, 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*1.
Example 41 (Exemp. compound No. 545)

In a similar manner to that described in Example 25 except for the use of 2-methyJdodecanoyl chloride [synthesized by chlorinating 2-methyldodecanoic acid

- 140-
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-dimethyldodecanoy] 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:
5 = 7.96 (d, J = 8.1 Hz, 1H), 6.01 (d, J = 4.0 Hz, 1H), 5.98 (dd, J = 4.5 and 3.4 Hz, 1H), 5.71 (d, } = 8.1 Hz, 1H), 5.46-5.43 (m, 1H), 5.24 (d, J - 5.5 Hz, 1H), 4.68 (d, 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 (t, J = 4.1 Hz, 1H), 4.08-4.05 (m, 1H), 3.97 (t, J = 5.0 Hz, 1H), 3.61-3.54 (m, 1H), 3.38 (s, 3H), 2.53-2.48 (m, 1H), 2.04-1.37 (m, 6H), 1.28 (s, 18H), 1.22 (d, J =6.6 Hz, 3H), 1.15-1.13 (m, 3H), 0.89 (t, 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*1.
Example 42 (Exemp. compound No. 571)
OH

O
CONH2 f^f-

-oA^yNYNH

In a similar manner to that described in Example 40 except for the use of 2-methyldodecanoyl chloride instead of 2,2-dimethyldodecanoyl chloride, 77.5 mgof 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:
5 = 7.95 (d, J = 8.1 Hz, 1H), 6.01 (d, J * 3.7 Hz, 1H), 5.98 (dd, J = 4.5 and 3.6 Hz, 1H), 5.72 (d, J = 8.1 Hz, 1H), 5.44-5.40 (m, 1H), 5.24 (d, J = 5.5 Hz, 1H), 4.68 (d, J = 1.8 Hz, 1H), 4.57-4.52 (m, 2H), 4.42 (t, J = 4.1 Hz, 1H), 4.04 (t, J = 4.8 Hz, 1H), 3.98

-142-
(t,J=5.0Hz, 1H), 3.37 (s,3H), 3.29-3.23 (m,2H), 2.23-2.48 (m,lH), 2,03-1.99 2) Infrared absorption spectrum: The Infrared absorption spectrum as measured by the potassium bromide (KBr) disk method exhfcte absorption maxima as follows: 3369,29277 2854,1689,1509,1461, 1382,1333,1269,1144,1110,1062 cm1.

-143-Test 1. Antibacterial activity
(1) Minimum inhbitor concentration
The minrnum inhfottory concentration of the compounds of the rivention aganst Mycobacterium smegmatis Strain SANK 75075 was determined in accordance with the process described below. The concentration of the compound to be tested was sest at four stages by four-fold dilution starting from 1000 fig/rnl
(1000 ug/ml, 250 M0/ml, 62 yg/ml and 15 jjgAn!). A 1 mlnportton of the diluted sample of each stage was poured into a ffetri dish ("Terumo Rstri disch", 90 x 20 mm). A nutrient agar medium (9 ml, product of Efcen Chemical) contefriing 5% gfycerol was added and they were mixed to prepare a plate medium. A test microorganism Mycob&cterium smegmatis SANK 75075 was precuttured overnight at 37°C on a trypto-soy broth (T.S.8) medium (product of Eicen Chemical) containng 5% glycerol. On the testing day, the microorganism solution was diluted 100-fold with T.S.B. and one toopful of the diluted culture was streaked onto the plate medium. After cultivation at 37°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.

144
Table 6
Antibacterial activities against
Mycobacterium smegmaiis SANtC 75075

The minimum inhibitory concentration of the invention compound of the formula (la) against Mycobacterium avium Strain N1HJ1605 was determined. Described specifically, Tween 80 (0.1%) was added to Middleblook 7H9 broth. After autoclave sterilization, Middleblook ADC enrichment was added (20%). Into each of micro-test 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 twofold (which will hereinafter be abbreviated as "medicament-containing medium"). On the side, a colony obtained by preculturing Mycobacterium avium

-145-
NIHJ1605 on a Tween egg medium for 10 to 14 days was charged in a test tube centering Tween 80 and glass beads. After sufficient mixng, Middlebknk 7H9 broth was added to form a uniform microorganism solution. The microorganism solution was adjusted to OD625nm=0.10 (viable cell count: about 1 x if/ CFU/ml), followed by 100-fold dilution. A 0.1 ml portion of the resulting microorganism solution was inoculated into the above-desabed medicament -containing medium (final viable cell count: about 1 x 105 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 orgreater was recognized on the bottom of the test tube was determned as MIC ( g/ml). The results are shown in Table 7.

(2) Disk Assay
So-called disk assay was conducted using 40 pa of a test substance per paper disk of 8 mm. Compound A-5003S9M-2 (Exemp. Compound No. 396) exhfcited an Inhbitory zone of 14 mm h a diameter against Bacillus subtiiis PCI219, that of 30 mm In a diameter against Myccfcacterlum smegrnatls SANK 75075 and that of 25 mm In diameter against Klebslella pneumonlae PCI602.

-146-

A capsule was obtained by mixing powders In accordance with the above-descrbed formulation, sieving the resulting mixture through a GO-mesh sieve, and then charging the resulting powder In a gelatin capsule.
Toxicity Test
The invention compound A-500359A exbfcrted no toxicrty when riiravenousty administEfed to a mouse in an amoifit of 500 mg/kg.
The results deserted above show that the compounds of the invention represented by the formula (I) and (la), and pharmacologically acceptable salts thereof exhibit excellent antibacterial activities against various bacterial Including Mycobacterta so that they are useful (n the prevention or treatment of hfecttous diseases caused by such bacteria. Streptomyces grteeus SANK60196 (FERM BP-5420) Is useful as a bacterium producJng the compound represented by the formula. The compounds represented by the formulae 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 mkrobiotogicat conversion.

147
WE CLAIM:
1. A compound of formula (I) or a pharmaceuticalIy acceptable salt thereof:

wherein:
R1 is a methyl group, R2 is a methyl group, R4 is a hydroxy group, and X is a methylene group;
R1 is a methyl group, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R4 is a hydrogen atom, and X is a methylene group;
R1 is a hydrogen atom, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a methylene group; or
R1 is a methyl group, R2 is a methyl group, R4 is a hydroxy group, and X is a sulfur atom.
2. A compound according to claim 1 or a pharmaceutically acceptable salt
thereof, wherein R1 is a methyl group, R2 is a methyl group, R is a hydroxy group,
and X is a methylene group.
3. A compound according to claim I or a phannaceutically acceptable salt
thereof, wherein the compound is of the formula (Ib):

and:

148
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a hydrogen atom, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
Rl is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydrogen atom, R5a is a hydrogen atom and X is a methylene group; or
R1 is a hydrogen atom, R2 is a hydrogen atom, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group.
4. A pharmaceutically acceptable ester or ether compound of formula (Ia) or N-alkylcarbamoyl derivative compound of formula (Ik), or a pharmaceutically acceptable salt thereof:

wherein:
R1 is a hydrogen atom or a methyl group;
R a is a hydrogen atom or a methyl group;
R3 is a hydrogen atom, a C6-20-alkylcarbonyl group, a C6-20-alkyloxycarbonyl group, a C10-20-alkenylcarbonyl group having 1 to 3 double bonds, or a C6-20-alkyl group;

149
R4a is a hydrogen atom or a hydroxy group;
R5 is a hydrogen atom, a C6-20-alkylcarbonyl group, a C6-20-alkyloxycarbonyl group, or a C10-20-alkenylcarbonyl group having 1 to 3 double bonds; R11 is a C1-21-alkyl group; and X is a methylene group or a sulfur atom,
PROVIDED THAT:
in formula (Ia) one or two ester residues are present as one or two of -OR and -OR5, or one ether residue is present as -OR3, or a combination thereof; AND THAT: when X is a sulfur atom,
R1 is a methyl group, R2a is a methyl group and R4a is a hydroxy group; when X is a methylene group, R1 is a methyl group and R a is a hydrogen atom,
R4a is a hydroxy group; or
when X is a methylene group and R1 is a hydrogen atom, R2a is a methyl group, and R4a is a hydroxy group.
5. A compound according to claim 4 or a pharmaceutically acceptable salt
thereof, wherein the compound is an ester compound of formula (Ia).
6. A compound according to claim 4 or a pharmaceuticaUy acceptable salt
thereof, wherein the compound is an ether compound of formula (Ia).
7. A compound according to claim 4 or a pharmaceutically acceptable salt
thereof, wherein the compound is an ester or ether compound of the formula (Ib):


150
and:
R1 is a methyl group, R2 is a methyl group, R3a is a decanoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R a is a lauroyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a myristoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a pentadecanoyl group, R a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a palmitoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R! is a hydrogen atom, R2 is a methyl group, R3a is a decanoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a lauroyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a myristoyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a pentadecanoyl group, R a i& >\ hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a palmitoyl group, R a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decanoyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a lauroyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R a is a myristoyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a pentadecanoyl group and X is a methylene group;
R is a methyl group, R is a methyl group, R a is a hydrogen atom, R ais a hydroxy group, R5a is a palmitoyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R a is a hydroxy group, R a is a decanoyl group and X is a methylene group;


R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a lauroyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a myristoyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a pentadecanoyl group and X is a methylene group;
R is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a palmitoyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hexyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a heptyloxycarbonyl group, R a is a hydroxy group, R a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is an octyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R is a methyl group, R is a methyl group, R a is a nonyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R is a methyl group, R is a methyl group, R a is a decyloxycarbonyl group, R a is a hydroxy group, R a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is an undecyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a do decyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R32 is a hydrogen atom, R4a is a hydroxy group, R a is a hexyloxycarbonyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R a is a heptyloxycarbonyl group and X is a methylene group;
R is a methyl group, R is a methyl group, R a is a hydrogen atom, R a is a hydroxy group, R5a is an octyloxycarbonyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a nonyloxycarbonyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a decyloxycarbonyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is an undecyloxycarbonyl group and X is a methylene group;

152
R1 is a methyl group, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a dodecyloxycarbonyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hexyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a heptyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is an octyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
Rl is a hydrogen atom, R2 is a methyl group, R3a is a nonyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a decyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is an undecyloxycarbonyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3E is a dodecyloxycarbonyl group, R"a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, Rsa is a hexyloxycarbonyl group and X is a methylene group;
R! is a hydrogen atom, R2 is a methyl group, R33 is a hydrogen atom, R4a is a hydroxy group, R5* is a heptyloxycarbonyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R a is an octyloxycarbonyl group and X is a methylene group;
R! is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4ais a hydroxy group, R5a is a nonyloxycarbonyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R3a is a decyloxycarbonyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is an undecyloxycarbonyl group and X is a methylene group;
R1 is a hydrogen atom, R2 is a methyl group, R3a is a hydrogen atom, R4a is a hydroxy group, R5a is a dodecyloxycarbonyl group and X is a methylene group;
R1 is a methyl group, R2 is a methyl group, R3a is a decyl group, R"a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group; or
R' is a hydrogen atom, R2 is a methyl group, R3a is a decyl group, R4a is a hydroxy group, R5a is a hydrogen atom and X is a methylene group.

153
8. A compound according to claim 4 or a pharmaceutically acceptable salt
thereof, wherein the compound is an N-alkylcarbamoyl derivative compound of
formula (Ik).
9. A compound according to claim 8 or a pharmaceutically acceptable salt
thereof, wherein:
R1 is a methyl group, R11 is a methyl group, R3 is a hydrogen atom and R5 is a hydrogen atom;
R1 is a methyl group, R11 is a methyl group, R3 is a decanoyl group and R5 is a hydrogen atom;
R1 is a methyl group, R11 is a methyl group, R3 is a hydrogen atom and R5 is a decanoyl group;
R1 is a methyl group, R11 is a dodecyl group, R3 is a hydrogen atom and R5 is a hydrogen atom;
R1 is a hydrogen atom, R11 is a methyl group, R3 is a hydrogen atom and R5 is a hydrogen atom;
R1 is a hydrogen atom, R11 is a methyl group, R3 is a decanoyl group and R5 is a hydrogen atom;
R1 is a hydrogen atom, R11 is a methyl group, R3 is a hydrogen atom and R5 is a decanoyl group; or
R1 is a hydrogen atom, R11 is a dodecyl group, R3 is a hydrogen atom and R5 is a hydrogen atom.
10. 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 9 or a pharmaceutically acceptable salt thereof.

154
11. A compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a bacterial infection.
12 . Slreptomyces griseus SANK60196 (FERM BP-5420).


A compound of formula (I) or a pharmaceutically acceptable salt thereof:


wherein: R1 is a methyl group, R2 is s methyl group, R4
is a hydroxy group , and X is a methylene group; R1 is a methyl

group, R2 is a hydrogen atom, R4 is a hydroxy group, and X is

a methylene group; R1 is a methyl group, R2 is a methyl group,

R4 is a hydrogen atom, and X is a methylene group; R1 is a hydrogen

atom, R2 is a hydrogen atom, R4 is a hydroxy group, and X is a

methylene group; or R1 is a methyl group, R2 is a methyl group,
R4 is a hydroxy group, and X is a sulfur atom.

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Patent Number 212615
Indian Patent Application Number IN/PCT/2001/00033/KOL
PG Journal Number 49/2007
Publication Date 07-Dec-2007
Grant Date 04-Dec-2007
Date of Filing 09-Jan-2001
Name of Patentee SANKYO COMPANY LIMITED
Applicant Address 5-1,NIHONBASHI HONCHO 3-CHOME, CHUO-KU,TOKYO 103-8426,JAPAN.
Inventors:
# Inventor's Name Inventor's Address
1 OGAWA YASUMASA, C/O.SANKYO COMPANY LIMITED, 389-4 AZA OHTSURUGI,IZUMICHOSHIMOKAWA,IWAKI-SHI,FUKUSHIMA 971-8183,JAPAN.
2 INUKAI MASATOSHI C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
3 KIZUKA MASAAKI C/O.SANKYO COMPANY LIMITED 33 MIYUKIGAOKA,TSUKUBA-SHI,IBARAKI 305-0841,JAPAN.
4 KANEKO MASAKATSU C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
5 HOTODA HITOSHI C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
6 MIYAKOSHI SHUNICHI C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
7 TAKATSU TOSHIO C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
8 ARAI MASATOSHI C/O.SANKYO COMPANY LIMITED, 2-58 HIROMACHI 1-CHOME, SHINAGAWA-KU, TOKYO 140-8710, JAPAN.
PCT International Classification Number C 07H 19/067
PCT International Application Number PCT/JP99/03718
PCT International Filing date 1999-07-09
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10/269445 1998-09-24 Japan
2 10/194285 1998-07-09 Japan