Note: Descriptions are shown in the official language in which they were submitted.
CA 02248784 1998-09-11
W O 97/49703 PCTIJP97/02157
DESCRIPTION
OXAZOLONE DERlVAlTVES AND THE~ USE AS ANnn-HE~COBAC7ER PYLORI AGENT
Technical Field
- The present invention relates to an anti-Helicobacter
pylori agent comprising an oxazolin-4-one derivative useful
as a therapeutic agent for gastric and duodenal ulcer etc.
Backqround Art
As therapeutic agents for ulcer, there have been
developed antacids, anticholinergic agents, antigastrin
agents, gastrointestinal hormones, antipepsine agents,
histamine H2 receptor antagonists, tissue repairing agents,
mucosa-protecting agents, microcirculation-improving
agents, proton pump inhibitors etc. The development of
histamine H2 receptor antagonists and proton pump
inhibitors, both possessing potent acid secretion-
suppressing activity, in particular, has facilitated ulcer
treatment.
However, these therapeutic agents for ulcer are
unsatisfactory in terms of suppressing effect on recurrent
ulcer. On the other hand, Helicobacter pylori, a gram-negati~e
microaerophilic bacterium belonging to the genus
Helicobacter, has been suggested as a potential major cause
of recurrence of gastritis, duodenal ulcer, gastric ulcer
etc. Although many antibacterial agents readily inhibit
the proliferation of the respective microorganisms
belonging to the genus HeZicobacterinuitro, their efficacy in
humans and animal experiments is very low when administered
singly in uiuo.
Various diseases caused by Helicobacterpylori as such are
treated by chemotherapies such as double chemotherapy with
a bismuth preparation and an antibiotic, and triple
chemotherapy with a bismuth preparation, metronidazole (US
Patent No. 2,944,061) and either tetracycline (e.g., US
CA 02248784 1998-09-11
- W O 97/49703 PCT/JP97/02157 - -
Patent No. 2,~12,517) or amoxicillin (US Patent No.
3,192,198). Metronidazole, an imidazole derivative
possessing anti-Helicobacterpylori activity, is used in
combination with antibiotics. These bismuth preparations,
antibiotics, metronidazole etc. are administered orally.
Also, clinical studies have shown that eradication of this
microorganism results in healing and decreased recurrence
rates in ulcer.
~owever, these bismuth preparations, antibiotics,
metronidazole etc. must be administered at high daily doses
to maintain sufficient concentrations to inhibit Helicobacter
pylori proliferation at the sites of their proliferation,
resulting in many problems, including adverse effects such
as vomiting and diarrhea.
There have been developed various compounds possessing
anti-Nelicobacterpylori activity. For example, Japanese
Patent Unexamined Publication No. 117268/1993 discloses a
pyridine derivative possessing anti-Helicobacterpylori
activity, and European Patent EPO 535528Al discloses an
imidazole derivative possessing anti-Helicobacterpylori
activity.
Disclosure of Invention
After extensive investigation in view of the above
problems, the present inventors found that a particular
oxazolin-4-one derivative exhibits very specific and
excellent antibacterial activity against the bacteria of
the genus Helicobacter, represented by Nelicobacterpylori. The
inventors conducted further investigation based on this
finding, and developed the present invention.
Accordingly, the present invention relates to
(1) an anti-Helicobacterpylori composition comprising a
compound of the formula:
CA 02248784 1998-09-11
W O 97/49703 PCT1JP97/02157
Rl R2 o
A ~ (I)
~ ~ ,R3
~ 5 R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3
and R4 independently represent a hydrogen atom, a
hydrocarbon group which may be substituted, an acyl group,
a carbamoyl group which may be substituted, or a carboxyl
group which may be esterified, or a salt thereof, and a
pharmacologically acceptable diluent, excipient or carrier,
(2) the anti-Helicobacterpylori composition according to the
description in (l) above, wherein A is an aromatic
heterocyclic group which may be substituted,
(3) the anti-Helicobacterpylori composition according to the
description in (1) above, wherein A is a group represented
by the formula:
~ (II)
wherein ring B is a 6-membered aromatic ring which may be
substituted, X represents CH or N, Y represents O, S or -N-
R5 (R5 represents a hydrogen atom or a hydrocarbon group
which may be substituted),
(4) the anti-Helicobacterpylori composition according to the
description in (1) above, wherein A is a group represented
by the formula:
(II-l)
Rs
CA 02248784 1998-09-11
- W097/49703 PCT/~97/02157
wherein ring B is a 6-membered aromatic ring which may be
substituted, R5 represents a hydrogen atom or a hydrocarbon
group which may be substituted,
(5) the anti-Helicobacterpylori composition according to the
description in (l) above, wherein A represented indolyl
which may be substituted by l to 3 substituents selected
from the group consisting of hydroxyl, halogen, nitro,
cyano, lower alkyl which may be substituted by l to 5
halogens and lower alkoxy which may be substituted by 1 to
5 halogens, Rl and R2 independently represent hydrogen or
lower alkyl which may be substituted by l to 5 halogens, R3
and R4 independently represent hydrogen or lower alkyl,
(6) the anti-Helicobacterpylori composition according to the
description in (5) above, wherein A is indolyl, Rl and R3
are hydrogen, and R2 and R4 are Cl_7 alkyl,
(7) the anti-Helicobacterpylori composition according to the
description in (6) above, wherein A is 3-indolyl, R2 and R4
are methyl,
(8) the anti-Helicobacterpylori composition according to the
description in (1) above, wherein the compound is
indolmycin,
(9) the anti-Helicobacterpylori composition according to the
description in (l) above, as an agent for prevention or
treatment of a disease associated with Helicobacterpylori
infection~
(lO) the anti-Helicobacterpylori composition according to the
description in (9) above, wherein the disease associated
with Helicobacterpylori infection is gastric or duodenal
ulcer, gastritis or gastric cancer,
(ll) the anti-Helicobacterpylori composition according to the
description in (l) above, which is used in combination with
an antibacterial agent,
(12) the anti-Helicobacterpylori composition according to the
description in (l) above, which is used in combination with
antiulcer agent,
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157
( 13 ) the anti-Helicobacterpylori composition according to the
description in (l) above, which is used in combination with
antibacterial agent and antiulcer agent,
(14) use of compound of the formula:
Rl R2 0
A ~ (I)
N~R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3
and R4 independently represent a hydrogen atom, a
hydrocarbon group which may be substituted, an acyl group,
a carbamoyl group which may be substituted, or a carboxyl
group which may be esterified, or a salt thereof for the
preparation of an anti-Helicobacterpylori agent,
(15) a method for prevention or treatment of a disease
associated with Helicobacterpylori infection in a mammal which
comprises administering to a subject in need an effective
amount of a compound of the formula (I):
Rl R2 o
A ~ (I)
,R3
N~R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3
and R4 independently represent a hydrogen atom, a
hydrocarbon group which may be substituted, an acyl group,
a carbamoyl group which may be substituted, or a carboxyl
group which may be esterified, or a salt thereof,
CA 02248784 1998-09-11
Wo 97/49703 PCT/lP97tO2157
( 16 ) a method for producing an anti-Helicobacterpylori
composition comprising mixing a compound of the formula
(I):
Rl R2 o
A ~ (I)
N~R3
R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3
and R4 independently represent a hydrogen atom, a
hydrocarbon group which may be substituted, an acyl group,
a carbamoyl group which may be substituted, or a carboxyl
group which may be esterified, or a salt thereof with a
pharmacologically acceptable diluent, excipient or/and
carrier,
(17) a compound of the formula:
Rl R2 o
A ~ (I')
~ ~ ,R3'
R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3'
and R4' independently represent a hydrogen atom or a
hydrocarbon group which may be substituted, or a salt
thereof, provided that ( 1 ) when A is 3-indolyl, Rl and R3 '
are hydrogen and R2 is methyl, R4 ' is neither C3-6
cycloalkyl nor mono-substituted Cl-4 alkyl wherein said
substituent is selected from halogen, hydroxyl, lower
alkoxy, lower thioalkyl, aryl, or an unsaturated 2-4 carbon
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157
atoms side-chain and (2) when A is 3-indolyl, Rl and R3 '
are hydrogen and R2 is Cl-3 alkyl, R4 ' is not selected from
hydrogen, phenyl, anisyl, toluidyl and C1_4 alkyl,
(18) a compound of the formula:
R1 R2 O
A ~ (I")
~ N~ R3
R4
wherein A represents an aromatic group which may be
substituted, Rl and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3'
is a hydrogen atom or a hydrocarbon group which may be
lS substituted, R4~ is an acyl group or a carbamoyl group
which may be substituted, or a salt thereof, provided that
when A is 3-indolyl, R1 is hydrogen and R2 and R3 ' are
methyl, R4~ is neither a C2_5 alkanoyl or an mono-
substituted C2_s alkanoyl wherein said substituent is
selected from amino, halogen, phenyl, p-hydroxyphenyl, or
lower alkoxy, nor a carbamoyl group substituted by C1_4
alkyl, C3-6 cycloalkyl or phenyl,
(l9) a compound of the formula:
Rl R2 o
A ~ (I
O--YN~ R3
R4 N
wherein A represents an aromatic group which may be
substituted, R1 and R2 independently represent a hydrogen
atom or a hydrocarbon group which may be substituted, R3 '
is a hydrogen atom or a hydrocarbon group which may be
substituted, R4n' is a carboxyl group which may be
esterified, or a salt thereof,
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97102157
(20) the compound according to the description in (19)
above, wherein A is indolyl, Rl and R2 independently
represent a hydrogen atom or methyl, R3' is methyl and R4n'
is a carboxyl group which is esterified,
(21) a method of producing indolmycin by culturing the
Streptomyces sp. HC-21 strain in a medium to produce and
accumulate indolmycin in the culture broth, and harvesting
the indolmycin, and
t22) the Streptomyces sp. HC-21 strain which assimilates L-
rhamnose and whose spores have a spiny surface.
The "aromatic ring group which may be substituted"represented by A in formula (I) is exemplified by
monocyclic or condensed polycyclic aromatic hydrocarbon
groups or aromatic heterocyclic groups. Such aromatic
hydrocarbon groups include, for example, phenyl, naphthyl,
anthryl, phenanthryl and acenaphthylenyl, with preference
given to phenyl, l-naphthyl, 2-naphthyl etc.
The aromatic heterocyclic groups include, for example,
aromatic monocyclic heterocyclic groups such as furyl,
thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, imidazolyl, pyrazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, furazanyl, 1,2,3-
thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,
1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyridyl,
pyridazinyl, pyrimidinyl, pyrazinyl and triazinyl; and
aromatic condensed heterocyclic groups such as
benzofuranyl, isobenzofuranyl, benzo[b]thienyl, indolyl,
isoindolyl, lH-indazolyl, benzimidazolyl, benzoxazolyl,
1,2-benzisoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl,
lH-benzotriazolyl, quinolyl, isoquinolyl, cinnolinyl,
quinazolinyl, quinoxalinyl, phthalazinyl, naphthyridinyl,
purinyl, pteridinyl, carbazolyl, a-carbolinyl, ~-
carbolinyl, y-carbolinyl~ acridinyl, phenoxazinyl,
phenothiazinyl, phenazinyl, phenoxathiinyl, thianthrenyl,
phenanthridinyl, phenanthrolinyl, indolizinyl, pyrrolo~l,2-
CA 02248784 1998-09-11
wo97l4s7o3 PCTl~971021S7- -
b]pyridazinyl, pyrazolo[l,5-alpyridyl, imidazo[l,2-
a]pyridyl, imidazo[l,5-a]pyridyl, imidazo[l,2-
b]pyridazinyl, imidazo[l,2-a]pyrimidinyl, 1,2,4-
triazolo[4,3-a~pyridyl and 1,2,4-triazolo[4,3-
b]pyridazinyl. Among aromatic condensed heterocyclicgroup, indolyl is preferable and 3-indolyl is more
preferable.
Substituents for the "aromatic ring group or aromatic
heterocyclic group which may be substituted" represented by
A in formula (I) include, for example, hydroxyl group,
halogens (e.g., fluorine, chlorine, bromine, iodine),
nitro, cyano, lower alkyls that may be substituted by 1 to
5 halogens (e.g., fluorine, chlorine, bromine, iodine),
lower alkoxys that may be substituted by 1 to 5 halogens
(e.g., fluorine, chlorine, bromine, iodine) benzyloxy and
Cl_4 alkoxy carbonyl (e.g. methoxy carbonyl, ethoxy
carbony, propoxy carbonyl, butoxy carbonyl). Such lower
alkyls include, for example, alkyl groups having 1 to 4
carbon atoms, such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl, sec-butyl and tert-butyl, wîth
preference given to methyl and ethyl. Such lower alkoxys
include alkoxy groups having 1 to 4 carbon atoms, such as
methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,
isobutoxy, sec-butoxy and tert-butoxy, with preference
given to methoxy and ethoxy. It is preferable that 1 to 3
(preferably 1 to 2) of these substituents, whether
identical or not, be present. Substituents for the
"aromatic ring group or aromatic heterocyclic group which
may be substituted" represented by A also include alkylene
dioxo such as methylene dioxo and ethylene dioxo.
The "hydrocarbon group which may be substituted"
represented by Rl or R2 in formula (I) include aliphatic
chain hydrocarbon groups, alicyclic hydrocarbon groups and
aryl groups, with preference given to aliphatic chain
hydrocarbon groups.
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157 --
Such aliphatic chain hydrocarbon groups include linear
or branched aliphatic hydrocarbon groups such as alkyl
groups, alkenyl groups and alkynyl groups. Particularly
preferred are lower alkyl groups, lower alkenyl groups,
lower alkynyl groups etc- Such lower alkyls include, for
example, Cl-7 alkyls such as methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-
pentyl, isopentyl, neopentyl, l-methylpropyl, n-hexyl,
isohexyl, l,l-dimethylbutyl, 2,2-dimethylbutyl, 3,3-
dimethylbutyl, 3,3-dimethylpropyl, 2-ethylbutyl and n-
heptyl. Preferred are Cl 3 alkyls such as methyl, ethyl
and propyl, with greater preference given to Cl_2 alkyls
such as methyl and ethyl. Such lower alkenyl groups
include, for example, C2_6 alkenyl groups such as vinyl,
allyl, isopropenyl, 2-methylallyl, l-propenyl, 2-methyl-1-
propenyl, 2-methyl-2-propenyl, l-butenyl, 2-butenyl, 3-
butenyl, 2-ethyl-1-butenyl, 2-methyl-2-butenyl, 3-methyl-2-
butenyl, l-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 4-
methyl-3-pentenyl, l-hexenyl, 2-hexenyl, 3-hexenyl, 4-
hexenyl and 5-hexenyl, with preference given to C2_s
alkenyls such as vinyl, allyl, isopropenyl, 2-methylallyl,
2-methyl-1-propenyl, 2-methyl-2-propenyl and 3-methyl-2-
butenyl. Such lower alkynyl groups include, for example,
C2_6 alkynyls such as ethynyl, l-propynyl, 2-propynyl, 1-
butynyl, 2-butynyl, 3-butynyl, l-pentynyl, 2-pentynyl, 3-
pentynyl, 4-pentynyl, l-hexynyl, 2-hexynyl, 3-hexynyl, 4-
hexynyl and 5-hexynyl, with preference given to C2_4
alkynyls such as ethynyl, l-propynyl and 2-propynyl.
Such alicyclic hydrocarbon groups include saturated or
unsaturated alicyclic hydrocarbon groups such as cycloalkyl
groups, cycloalkenyl groups and cycloalkadienyl groups.
Such cycloalkyl groups are preferably cycloalkyl groups
having 3 to 9 carbon atoms, such as cyclopropyl,
3~ cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl and cyclononyl, with greater preference given to
CA 02248784 1998-09-11
W O 97/49703 PCTIJP97/02157 - --
C3-6 cycloalkyl groups such as cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl. Such cycloalkenyl groups
include, for example, C3-6 cycloalkenyls such as 2-
cyclopenten-l-yl, 3-cyclopenten-1-yl, 2-cyclohexen-1-yl, 3-
cyclohexen-l-yl, l-cyclobuten-l-yl and l-cyclopenten-l-yl.
Such cycloalkadienyl groups include, for example, C4-6
cycloalkadienyls such as 2,4-cyclopentadien-1-yl, 2,4-
cyclohexadien-l-yl and 2,5-cyclohexadien-1-yl.
The aryl groups in the hydrocarbon groups include
monocyclic or condensed polycyclic aromatic hydrocarbon
groups such as phenyl, naphthyl, anthryl, phenanthryl and
acenaphthylenyl, with preference given to phenyl, 1-
naphthyl, 2-naphthyl etc.
Substituents for the "hydrocarbon group which may be
substituted" represented by Rl or R2 in formula (I) include
aryl groups which may be substituted, cycloalkyl or
cycloalkenyl groups which may be substituted, heterocyclic
groups that may be substituted, amino group that may be
substituted, hydroxyl group which may be substituted, thiol
group which may be substituted, and halogens (e.g.,
fluorine, chlorine, bromine, iodine). One to five
(preferably 1 to 3) of these optionally chosen substituents
may be present. Such aryl groups which may be substituted
include phenyl, naphthyl, anthryl, phenanthryl and
acenaphthylenyl, with preference given to phenyl, 1-
naphthyl and 2-naphthyl. Substituents for such aryl groups
which may be substituted include alkoxy groups having 1 to
3 carbon atoms (e.g., methoxy, ethoxy, propoxy), halogen
atoms (e.g., fluorine, chlorine, bromine, iodine) and alkyl
groups having 1 to 3 carbon atoms (e.g., methyl, ethyl,
propyl); 1 to 2 of these optionally chosen substituents may
be present. Such cycloalkyl groups which may be
substituted include C3_7 cycloalkyl groups such as
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and
cycloheptyl. The kinds and number of substituents for such
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157-
cycloalkyl groups which may be substituted are the same as
those for the substituents the above-described aryl group
that may be substituted. Such cycloalkenyl groups which
may be substituted include C3-6 cycloalkenyl groups such as
cyclopropanyl, cyclobutenyl, cyclopentenyl and
cyclohexenyl. The kinds and number of substituents for
such cycloalkenyl groups which may be substituted are the
same as those for the substituents for the above-described
aryl group that may be substituted. Such heterocyclic
groups which may be substituted for include aromatic
heterocyclic groups having at least l hetero atom selected
from oxygen, sulfur or nitrogen as a ring-constituting atom
(ring atom), and saturated or unsaturated non-aromatic
heterocyclic groups (aliphatic heterocyclic groups), with
preference given to aromatic heterocyclic groups. Such
aromatic heterocyclic groups include aromatic monocyclic
heterocyclic groups (e.g., furyl, thienyl, pyrrolyl,
oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl,
pyrazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-
oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-
thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-
triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl,
pyrazinyl, triazinyl) and aromatic condensed heterocyclic
groups (e.g., benzofuranyl, isobenzofuranyl,
benzo[b]thienyl, indolyl, isoindolyl, lH-indazolyl,
benzimidazolyl, benzoxazolyl, 1,2-benzisoxazolyl,
benzothiazolyl, 1,2-benzisothiazolyl, lH-benzotriazolyl,
quinolyl, isoquinolyl, cinnolinyl, quinazolinyl,
quinoxalinyl, phthalazinyl, naphthyridinyl, purinyl,
pteridinyl, carbazolyl, a-carbolinyl~ ~-carbolinyl, y-
carbolinyl, acridinyl, phenoxazinyl, phenothiazinyl,
phenazinyl, phenoxathiinyl, thianthrenyl, phenanthridinyl,
phenanthrolinyl, indolizinyl, pyrrolo[l,2-b]pyridazinyl,
pyrazolo[l,5-a]pyridyl, imidazo[l,2-a~pyridyl, imidazo[l,5-
a]pyridyl, imidazo[l,2-b3pyridazinyl, imidazo[l,2-
a]pyrimidinyl, 1,2,4-triazolo[4,3-a]pyridyl, 1,2,4-
CA 02248784 1998-09-11
W097/49703 PCT/~97/021S7
triazolo[4,3-b~pyridazinyl), with preference given to
furyl, thienyl, indolyl, isoindolyl, pyrazinyl, pyridyl,
pyrimidinyl etc. Such non-aromatic heterocyclic groups
include, for example, oxylanyl, azetidinyl, oxetanyl,
thietanyl, pyrrolidinyl, tetrahydrofuryl, thiolanyl,
piperidyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl
and piperazinyl. Substituents for such heterocyclic groups
which may be substituted for include alkyl groups having l
to 3 carbon atoms (e.g., methyl, ethyl, propyl).
Substituents for such amino group which may be substituted
for, hydroxyl group that may be substituted for, and thiol
group that may be substituted for, include, for example,
lower (Cl_3) alkyl groups (e.g., methyl, ethyl, propyl).
When the "hydrocarbon group which may be a substituted"
represented by Rl or R2 is an alicyclic hydrocarbon group
or an aryl group, the substituent may also be an alkyl
group having l to 3 carbon atoms (e.g., methyl, ethyl,
propyl).
With respect to the formula (I), the preferable
combination of Rl and R2 is that Rl is hydrogen and R2 is
Cl_3 alkyl which may be substituted by l to 5 halogens.
The hydrocarbon group and the substituent in the
"hydrocarbon group which may be substituted" represented by
R3 or R4 in formula (I) and represented by R3' or R4' in
formula (I') are exemplified by the same hydrocarbon groups
and substituents mentioned to exemplify the hydrocarbon
group and substituent for Rl and R2 above, respectively.
With respect to the formula (I), the preferable
combination of R3 and R4 is that R3 is hydrogen and R4 is
Cl_3 alkyl.
The acyl group represented by R3 or R4 in formula (I)
is exemplified by aliphatic acyl groups such as alkanoyl
groups, alkenoyl groups, cycloalkanecarbonyl groups and
alkanesulfonyl groups; aromatic acyl groups such as aroyl
groups, arylalkanoyl groups, arylalkenoyl groups and
CA 02248784 1998-09-11
wo97l4s7o3 PCT/~7/02157
14
arenesulfonyl groups; heterocyclic aromatic acyl groups
such as aromatic heterocyclic carbonyl groups and aromatic
heterocyclic alkanoyl groups; and non-aromatic heterocyclic
carbonyl groups (aliphatic heterocyclic carbonyl groups).
"Alkanoyl groups" mean alkylcarbonyl groups,
preferable examples thereof including lower alkanoyl groups
having l to 8 carbon atoms, such as formyl, acetyl,
propionyl, butyryl, isobutyryl, valeryl, isovaleryl,
pivaloyl and hexanoyl.
"Alkenoyl groups" mean alkenylcarbonyl groups,
preferable examples thereof including C3-6 alkenoy} groups
such as acryloyl, methacryloyl, crotonoyl and isocrotonoyl.
"Cycloalkanecarbonyl groups" mean cycloalkylcarbonyl
groups, preferable examples thereof including those having
4 to 7 carbon atoms, such as cyclopropanecarbonyl groups,
cyclobutanecarbonyl groups, cyclopentanecarbonyl groups and
cyclohexanecarbonyl groups.
"Alkanesulfonyl groups" mean alkylsulfonyl groups,
preferable examples thereof including those having 1 to 4
carbon atoms, such as mesyl, ethanesulfonyl and
propanesulfonyl.
"Aroyl groups" mean arylcarbonyl groups, preferable
examples thereof including those having 7 to ll carbon
atoms, such as benzoyl, p-toluoyl, l-naphthoyl and 2-
naphthoyl.
"Arylalkanoyl groups" mean alkylcarbonyl groups
substituted for by an aryl group, preferable examples
thereof including C6_g aryl-C2 5 alkanoyl groups such as
phenylacetyl, phenylpropionyl, hydroatropoyl and
phenylbutyryl.
"Arylalkenoyl groups" mean alkenylcarbonyl groups
substituted for by an aryl group, preferable examples
thereof including C6_g aryl-C3_s alkenoyl groups such as
cinnamoyl and atropoyl.
CA 022487X4 1998-09-11
W097/49703 PCT/~97/021S7- _
"Arenesulfonyl groups" mean arylsulfonyl groups,
preferable examples thereof including those having 6 to 8
carbon atoms, such as benzenesulfonyl and p-
toluenesulfonyl.
Preferable examples of "aromatic heterocyclic carbonyl
groups" include furoyl, thenoyl, nicotinoyl, isonicotinoyl,
pyrrolecarbonyl, oxazolecarbonyl, thiazolecarbonyl,
imidazolecarbonyl and pyrazolecarbonyl.
"Aromatic heterocyclic alkanoyl groups" mean
alkylcarbonyl groups substituted by an aromatic
heterocyclic group, preferable examples thereof including
aromatic heterocyclic ring-C2_s alkanoyl groups such as
thienylacetyl, thienylpropanoyl, furylacetyl,
thiazolylacetyl, l,2,4-thiadiazolylacetyl and
pyridylacetyl~
Preferable examples of "non-aromatic heterocyclic
carbonyl groups" include aliphatic heterocyclic carbonyls
such as azetidinylcarbonyl, pyrrolidinylcarbonyl and
piperidinylcarbonyl.
The carbamoyl group which may be substituted
represented by R3 or R4 in formula (I) and represented by
R4' in formula (I"), is exemplified by "N-
monosubstitutional carbamoyl groups" and "N,N-
disubstitutional carbamoyl groups," as well as non-
substitutional carbamoyl. An "N-monosubstitutional
carbamoyl group" means a carbamoyl group having one
substituent on nitrogen. Examples of said substituent
include Cl_6 alkyls (e.g., methyl, ethyl, propyl,
isopropyl, butyl, t-butyl, pentyl, hexyl), C3-6 cycloalkyl
groups (e.g., cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl), aryl groups (e.g., phenyl, l-naphthyl, 2-
naphthyl), aralkyl groups (e.g., benzyl, phenethyl) and
heterocyclic groups (e.g., the "heterocyclic groups"
mentioned to exemplify the "substituent" for the
"hydrocarbon residue which may be substituted" represented
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157 --
16
by Rl or R2 above). Such aryl groups, aralkyl groups and
heterocyclic groups may be substituted. Said substituent
is exemplified by hydroxyl group, amino group which may
substituted by l or 2 lower alkyls (e.g., those having l to
4 carbon atoms, such as methyl, ethyl, propyl, isopropyl
and butyl) or acyl groups (e.g., formyl, acetyl, propionyl,
benzoyl), halogens (e.g., fluorine, chlorine, bromine,
iodine), nitro, cyano, lower alkyls which may be
substituted by l to 5 halogens (e.g., fluorine, chlorine,
bromine, iodine) and lower alkoxys which may be substituted
by l to 5 halogens (e.g., fluorine, chlorine, bromine,
iodine). Such lower alkyls include, for example, alkyl
groups having l to 4 carbon atoms, such as methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-
butyl, with preference given to methyl and ethyl. Suchlower alkoxys include alkoxy groups having l to 4 carbon
atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-
butoxy, isobutoxy, sec-butoxy and tert-butoxy, with
preference given to methoxy and ethoxy. It is preferable
that l to 3 (preferably l to 2) of these substituents,
whether identical or not, be present.
An "N,N-disubstitutional carbamoyl group" means a
carbamoyl group having two substituents on a nitrogen atom.
Examples of one of said substituents include the same
substituents as those mentioned to exemplify the
substituent for the "N-monosubstitutional carbamoyl group"
above; examples of the other substituent include Cl-6 alkyl
groups (e.g., methyl, ethyl, propyl, isopropyl, butyl, t-
butyl, pentyl, hexyl), C3-6 cycloalkyl groups (e.g.,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl) and C6_l0
aralkyl groups (e.g., benzyl, phenethyl). The two
substituents may form a cyclic amino group in cooperation
with the nitrogen atom. In this case, examples of cyclic
aminocarbamoyl groups include l-azetidinylcarbonyl, l-
pyrrolidinylcarbonyl, piperidinocarbonyl,
CA 02248784 1998-09-11
- W097/4g703 PCT/~7lo2ls7 --
17
morpholinocarbonyl, l-piperazinylcarbonyl, and l-
piperazinylcarbonyl having a lower alkyl group such as Cl_6
alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, t-
butyl, pentyl, hexyl), an aralkyl group such as benzyl and
phenethyl, an aryl group such as phenyl, l-naphthyl and 2-
naphthyl, or the like, at the 4-position.
The "carboxyl group which may be esterified"
represented by R3 or R4 in formula (I) and represented by
R4n' in formula (I"'), is exemplified by "lower
alkoxycarbonyl groups," "aryloxycarbonyl groups" and
"aralkyloxycarbonyl groups," as well as free carboxyl
group.
Preferable examples of "lower alkoxycarbonyl groups"
include those having 2 to 8 carbon atoms, such as
methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,
isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-
butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl,
isopentyloxycarbonyl, neopentyloxycarbonyl and tert-
pentyloxycarbonyl, with preference given to those having 2to 4 carbon atoms, such as methoxycarbonyl, ethoxycarbonyl
and propoxycarbonyl.
Preferable examples of "aryloxycarbonyl groups"
include those having 7 to 12 carbon atoms, such as
phenoxycarbonyl, l-naphthoxycarbonyl and 2-
naphthoxycarbonyl. Preferable examples of
"aralkyloxycarbonyl groups" include those having 8 to lO
carbon atoms, such as benzyloxycarbonyl and
phenethyloxycarbonyl. These aryloxycarbonyl groups and
aralkyloxycarbonyl groups may be substituted; useful
substituents are identical to those mentioned to exemplify
the substituent for aryl groups and aralkyl groups in the
case of N-monosubstitutional carbamoyl groups.
The "6-membered aromatic ring which may be
substituted" represented by ring B in formula (II) is
CA 02248784 1998-09-11
- WO 97/49703 PCT/JP97~2157 --
18
exemplified by benzene ring which may be substituted, and
6-membered aromatic heterocyclic ring that may be
substituted. When ring B represents a benzene ring which
may be substituted, formula (II) represents a group
represented by the formula:
~ (II-a)
wherein ring C may be substituted; X and Y have the same
definitions as those shown above. When ring B represents a
6-membered aromatic heterocyclic ring which may be
substituted, the groups represented by formula (II)
include, for example, those represented by the following
formulas:
~ ~ N ~ ~ , Y
N ~ N~ 3 ~ N ~ ~ ~ N
N ~ N~,D ~ ~D ~ ~ ~
In these formulas, ring D may be substituted; X and Y
have the same definitions as those shown above.
With respect to the above formulas, the substituents
for rings C and D are identical to those mentioned to
exemplify the "substituent" for the "aromatic ring group
which may be substituted", represented by A. These
substituents may bound to any carbon atom of rings C and D.
The "hydrocarbon group which may be substituted"
represented by R5 in formula (II-l) is exemplified by the
same hydrocarbon groups as those mentioned to exemplify the
CA 02248784 1998-09-11
W097/49703 PCT/~97/021S7- --
hydrocarbon group represented by Rl or R2, which may be
substituted.
With respect to the formula (I), the preferable
combination of A, Rl, R2, R3 and R4 is that A is indolyl,
Rl and R3 are hydrogen, and R2 and R4 are Cl-3 alkyl. The
specific examples of the above-described oxazolin-4-one
derivative include indolmycin.
Salts of the compound represented by formula (I),
(I'), (I") or (I"') include pharmacologically acceptable
acid addition salts; acids that form an acid addition salt
include acetic acid, lactic acid, succinic acid, maleic
acid, tartaric acid, citric acid, gluconic acid, ascorbic
acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic
acid, cinnamic acid, fumaric acid, phosphonic acid,
hydrochloric acid, hydrogen bromide, hydrogen iodide,
sulfamic acid and sulfonic acid.
Examples of the compound represented by the formula
(I) are given below.
CA 02248784 1998-09-11
- W097/49703 PCT/JP97/02157 --
Rl R2 o
A ~
~ ~R4
Compound A Rl R2 R3 R4
Number
1) ~ H H H H
H
2) ~ H H H CH3
3) ~ H H H CH2CH3
H H H H (CH2)2CH3
5) ~ H H H CH(CH3)2
6) ~ H H H
7) ~ H H H (CH2)3CH3
8) ~ H CH3 H H
9) ~ H CH3 H CH3
10) ~ H CH3 H CH2CH3
11) ~ H CH3 H (CH2)2CH3
12) ~ H CH3 H CH(CH3)2
H
CA 02248784 1998-09-11
W097t49703 PCTIJP97/02157 ~~
A ~
N~R3
Number A Rl R2 R3 R4
13) ~ H CH3 H
H
14) ~ H CH3 H (cH2)3cH3
15) ~ H CH3 H CH2CH(CH3)2
16) ~ H CH3 H
17) ~ H CH3 H CH
18) H H CH3 H (CH2)2
19) ~ H CH3 CH3 CH3
20) ~ H CH3CH2CH3 CH2CH3
21) H H CH3 H CH2CH2OH
22) ~ H CH3 H CH2CH2OCH3
23) ~ H CH3 H CH2CH2CN
24) ~ CH3 CH3 H CH3
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157 --
22
Rl R2 o
~ N~R3
Compound A RlR2 R3 R4
Number
25) ~ C~3 CH3 CH3 CH3
H
26) X HCH2CH3 H CH3
27) H HCH2CH3 H CH3
28) H H(CH2)2CH3 H CH3
29) H H(CH2)2CH3 CH3 CH3
30) H HCH(CH3)2 H CH3
31) H HCH(CH3)2 CH3 CH3
32) H H CH3 CH3COCH3
33) ~ H CH3 CH3COCH2CH3
34) ~ H CH3 CH3CO(CH2)2CH3
35) ( ~ H CH3 CH3COCH(CH3)2
3 6) H H CH3 CH3CO(CH2)3CH3
CA 02248784 1998-09-11
W O 9714g703 PCT/JPg7/02157 --
Rl R2 o
A ~
N~R4
Compound A RlR2 R3 R4
Number
37) ~ HCH3 CH3CO(CH2)4CH3
H
38) H HCH3 CH3CO(CH2)5CH3
39) ~ H CH3 CH3 CO
40) H H CH3 CH3 COCH2
41) ~ H CH3 CH3 CO(CH2)2
H
42) H H CH3 CH3 CO ~ Cl
43) ~ H CH3 CH3 CO ~ Br
44) H H CH3 CH3 CO ~ F
45) ~ H CH3 CH3 CO ~ CF3
46) H H CH3 CH3 CO ~ CH3
47) ~ H CH3 CH3 CO ~ OCH3
48) H H H H COCH3
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157 --
24
Rl R2 o
A ~
N~R4
Compound A RlR2 R3 R4
Number
49) ~ H H HCO ~ )
H
50) ~ H H HCONHCH3
51) ~ H H HCONHCH2CH3
52) ~ H H HCONH(CH2)2CH3
53) ~ H H HCONHCH(CH3)2
54) ~ H H H CONH(CH2)3CH3
55) ~ H H H CONH(CH2)5CH3
2 5 56) ~ N~ H H H CONH
57) ~ H H H CONHCH
58) ~ H H H CONH ~
s9) ~ H H H CONH ~ F
60) ~ H CH3 HCONHCH3
CA 02248784 1998-09-ll
W 097/49703 PCT/JP97/02157 --
Rl R2 o
A ~
~ ~R3
Compound A Rl R2 R3 R4
Numbe r
61) ~ H CH3 HCONHCH2CH3
H
6 2) H H CH3 HCONHtCH2)2CH3
63) H H CH3 HCONHCH(CH3)2
64) H H CH3 HCONH(CH2)3CH3
65) H H CH3 HCONH(CH2)5CH3
66) H H CH3 HCONH ~ )
67) H H CH3 HCONHCH
68) H H CH3 HCONH ~
69) ~ H CH3 HCONH ~ F
70) ( ~ H CH3CH3CONHCH3
71) H H CH3CH3CONHCH2CH3
72) H H CH3CH3CONHCH(CH3)2
CA 02248784 1998-09-11
W O 97t49703 PCT/JP97/02157 --
26
Rl R2 o
~ N~R3
R4
Number A Rl R2 R3 R4
73) ~ H CH3 CH3CONH(CH2)2CH3
H
74) ~ H CH3 CH3CONH(CH2)3CH3
75) ~ H CH3 CH3CONH(CH2)4CH3
76) ~ H CH3 CH3CONH(CH2)sCH3
77) ~ H CH3 CH3 CONH
~5
78) ~ H CH3 CH3 CONH
79) ~ H CH3 CH3 CONH ~
80) ~ H CH3 CH3 CONHCH2 ~ 9
81) h H CH3 CH3 CONH ~
82) ~ H CH3 CH3 CONH ~ Cl
83) ~ H CH3 CH3 CONH ~ Br
84) H H CH3 CH3 CONH ~ F
CA 02248784 1998-09-11
W097/49703 PCT/JP97/02157 ~~
Rl R2 o
X~
'R4
Compound
Number A Rl R2 R3 R4
85) E H CH3 CH3 CONH ~ CH3
86) E H CH3 CH3 CONH ~ OCH3
87) E H CH3 CH3 CONH ~ CF3
88) E CH3 CH3 CH3 CONHCH3
89) E CH3 CH3 CH3 CONHCH2CH3
90) E CH3 CH3 CH3 CONH ~
91) E CH3 CH3 CH3 CONH ~ Cl
92) E H H H CO
93) ~ H H H
H CO
94) ~ H H H CO
95) ~ H H H CO
96) E H H H CO
CA 02248784 1998-09-11
W O 97/49703 PCTIJP97/02157 _
A ~
N~R3
Number A Rl R2 R3 R4
97) ~ H H H ~
H CO N
98) H H H H CO ~ N
H H CH3 CH3 CO
lO0) H H CH3 CH3 CO
101) H H CH3 CH3 CO
102) H H CH3 CH3 CO
103) H H CH3 CH3 CO ~
104) E H CH3 CH3 CO ~ N
105) H H CH3 CH3 CO ~ N
106) ~ H CH3 CH
H COCH2 S
107) ~ H CH3 CH
H COCH2
108) H H CH3 CH3 CO~ CH3
CA 02248784 1998-09-11
Wog7/49703 PCT/JP97/02157
29
Rl R2 o
A ~
~ N~R3
R
Compound A Rl R2 R3 R4
Number
109) ~ H CH3 CH3 CO
H
110) ~ H CH3 CH3 CO
111) ~ H CH3 CH3 CO ~/
112) ~ H CH3 CH3CONH
113) ~ H CH3 CH3
H CONH
114) E H CH3 CH3CONH
115) ~ H CH3 CH3
H CONH
116) ~ H CH3 CH3CONHCH2
117) ~ H CH3 CH3
H CONHCH2
118) ~ H H H H
119) ~ H H H CO
120) ~ H H H CONH
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157
Rl R2 o
A ~
~R4
Compound A Rl R2 R3 R4
Number
121) ~ H CH3 CH3 H
122) 1 ~ H CH3 CH3 CO ~ 3
- 15 123) [ ~ H CH3 CH3 CO
12 4) ~ H CH3 CH3 CONH ~ 9
125) ~ ~ CH3 CH3 CONH
126) ~ H H H H
127) ~ H H H CO ~
128) ( ~ H H HCONH ~ 3
129) 1 ~ H CH3 CH3 H
130) ( ~ H CH3 CH3 CO
131) ~ H CH3 CH3 CO
132) ~ H CH3 CH3 CONH
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/021S7 --
R~ O
~ ~ 4
R
Compound A Rl R2 R3 R4
Num~er
133) ~ H CH3 CH3 CONH
134) ~ H H H H
135) ~ H H H CO
H
136) ~ H H H CONK
H
137) H H CH3 CH3 H
138) H H CH3 CH3 CO
139) H H CH3 CH3 CO
140) ~ H CH3 CH3
H CO
141) ~ H CH3 CH3 CONH
H
142) H H CH3 CH3 ~
143) ~ H CH3 CH3 CO ~ CH3
H
144) H H CH3 CH3 CO
-
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97tO2157
Rl R2 o
A ~
~ ~R3
Number A Rl R2 R3 R4
145) ~ H CH3 CH3 COOCH3
H
146) H H CH3 CH3COOCH2CH3
147) ~ H CH3 CH3COO(CH2)2CH3
148) H H CH3 CH3COOCH(CH3)2
149) H H CH3 CH3COO(CH3)3CH3
150) H H CH3 CH3COOCH2CH(CH3)
151) ~ H CH3 CH3COOC(CH3)3
152) ~ H CH3 CH3COO(CH2)4CH3
153) H H CH3 CH3COO(CH2)5CH3
154) H H CH3 CH3 COOCH
155) H H CH3 CH3 SO2CH3
~5 156) H H CH3 CH3SO2CH2CH3
CA 02248784 1998-09-ll
W O 97/49703 PCT/JP97/02157
Rl R2 o
A ~
O ~ R3
N~R4
Compound
Number A Rl R2 R3 R4
157) ~ H CH3 CH3SO2(CH2)2CH3
H
158) H H CH3 CH3SO2(CH2)3CH3
159) ~ H CH3 CH3SO2(CH2)4CH3
160) H H CH3 CH3SO2(CH2)sCH3
H
161) H H CH3 CH3 SO
162) H H CH3 CH3 SO
163) H H CH3 CH3 S02 ~
164) H H CH3 CH3 SO2 ~ C1
165) H H CH3 CH3 SO2 ~ CH3
166) H H CH3 CH3
The above compounds may be racemates or optical
isomers.
CA 02248784 1998-09-11
W097149703 PCT/~7102157- -
34
Compound (I) or a salt thereof, used for the present
invention, is effective as an antibacterial agent in the
prevention or treatment of "duodenal ulcer, gastric ulcer,
gastritis (including chronic gastritis), gastric cancer
etc." caused by Hel~cobacterpylori infection as described
above, because it possesses antibacterial activity,
especially potent antibacterial activity against the
bacteria of the genus Hel~cobacter, represented by Helicobacter
pylori .
The preparation of the present invention, containing
compound tI) or a pharmacologically acceptable salt
thereof, can be orally or non-orally administered as an
antibacterial or antiulcer agent to mammals (e.g., humans,
dogs, cats, monkeys, rats, mice, horses, bovines), oral
administration being normally preferred.
Examples of dosage forms for oral administration
include tablets (including sugar-coated tablets and film-
coated tablets), pills, granules, powders, capsules
(including soft capsules), syrups, emulsions and
suspensions. Examples of dosage forms for non-oral
administration include injectable preparations, infusions,
drip infusions and suppositories.
The content of compound (I) or a salt thereof in the
preparation of the present invention is normally 2 to 85%
by weight, preferably 5 to 70% by weight.
For preparing compound (I) or a salt thereof in the
above-mentioned dosage forms, known production methods in
common use in relevant fields are applicable. In producing
the above-mentioned dosage forms, excipients, binders,
disintegrants, lubricants, sweetening agents, surfactants,
suspending agents, emulsifiers etc. in common use in the
field of pharmaceutical making may be added in appropriate
amounts as necessary.
When compound (I) or a salt thereof is prepared as
tablets, for example, excipients, binders, disintegrants,
CA 02248784 1998-09-11
W097t49703 PCT/~97/02157- -
lubricants etc. may be contained; when compound (I) or a
salt thereof is prepared as pills or granules, excipients,
binders, disintegrants etc. may be contained. When
compound (I) or a salt thereof is prepared as powders or
capsules, excipients etc. may be contained; when compound
(I) or a salt thereof is prepared as syrups, sweetening
agents etc. may be contained; when compound (I) or a salt
thereof is prepared as emulsions or suspensions, suspending
agents, surfactants, emulsifiers etc. may be contained.
Examples of excipients include lactose, saccharose,
glucose, starch, sucrose, microcrystalline cellulose,
powdered glycyrrhiza, mannitol, sodium hydrogen carbonate,
calcium phosphate and calcium sulfate. Examples of binders
include 5-10% by weight starch glue solutions, 10-20~ by
weight gum arabic solutions or gelatin solutions, 1-5% by
weight tragacanth solutions, carboxymethyl cellulose
solutions, sodium alginate solutions and glycerol.
Examples of disintegrants include starch and calcium
carbonate. Examples of lubricants include magnesium
stearate, stearic acid, calcium stearate and purified ta~c.
Examples of sweetening agents include glucose, fructose,
invert sugar, sorbitol, xylitol, glycerol and simple
syrups. Examples of surfactants include sodium lauryl
sulfate, polysorbate 80, sorbitan monofatty acid ester and
stearic acid polyoxyl 40. Example of suspending agents
include gum arabic, sodium alginate, carboxymethyl
cellulose sodium, methyl cellulose and bentonite. Examples
of emulsifiers include gum arabic, tragacanth, gelatin and
polysorbate 80.
For preparing compound ~I) or a salt thereof in the
above-mentioned dosage forms, coloring agents,
preservatives, flavoring agents, correctives, stabilizers,
thickening agents etc. in common use in the field of
pharmaceutical making may be added in appropriate amounts
as desired. The preparation of the present invention,
CA 02248784 l998-09-ll
W O 97/49703 PCT/JP97/02157- - .
36
which contains a compound represented by general formula
(I) or a pharmaceutically acceptable salt thereof, is
stable and of low toxicity, and can be safely used.
Varying depending on patient condition and body weight,
kind of compound, route of administration etc., the daily
dose of the preparation of the present invention is
normally 1 to 500 mg, preferably about 10 to 200 mg, based
on active ingredient content (compound (I) or a salt
thereof), per adult (weighing about 60 kg) for oral
administration in patients with gastric ulcer caused by
Helicobacterpylori infection.
Within the above-described dose range, no toxicity is
seen.
Also, in the preparation of the present invention,
compound (I) or a salt thereof can be used in combination
with other antibacterial agents and antiulcer agents.
Other antibacterial agents that can be used in
combination with compound (I) or a salt thereof include,
for example, nitroimidazole antibiotics (e.g., tinidazole
and metronidazole), tetracyclines (e.g., tetracycline,
doxycycline and minocycline), penicillins (e.g.,
amoxicillin, ampicillin and mezlocillin), cephalosporins
(e.g., cefaclor, cefadroxil, cefazolin, cefuroxime,
cefuroxime axetil, cephalexin, cefpodoxime proxetil,
ceftazidime and ceftriaxone), carbapenems (e.g., imipenem
and meropenem), aminoglycosides ~e.g., paromomycin),
macrolide antibiotics (e.g., erythromycin, clarithromycin
and azithromycin), lincosamide antibiotics (e.g.,
clindamycin), rifamycins (e.g., rifampicin) and quinolone
antibiotics (e.g., ciprofloxacin, ofloxacin)
nitrofurantoin. Antiulcer agents that can be used in
combination with compound (I) or a salt thereof include,
for example, proton pump inhibitors (e.g.,~omeprazole~
lansoprazole,~pantoprazole, rabeprazole) Histamine H2
antagonists (e.g., ranitidine, cimetidine and famotidine),
CA 02248784 l998-09-ll
W 097/49703 PCT/JP97/021S7
and mucosa-protecting antiulcer agents (e.g., sofalcone,
plaunotol, teprenone, sucralfate).
The above-described other antibacterial agents and
antiulcer agents may be uged in combination of two or more
kinds. In this case, the dose of antibacterial agent is
normally 1 to 500 mg, preferably 5 to 200 mg, per adult per
day in oral administration; the dose of antiulcer agent is
normally 0.5 to 1,000 mg, preferably 1 to 500 mg, per adult
per day in oral administration.
The compound of formula (I) or a salt thereof can, for
example, be produced by methods A through E below.
Method A
o
~ N
RlX2 --~N
A Z R4
(IV)
(III) Base ~ (I)
In the above formulas, Z represents a halogen atom or
-O-SO2R6 (R6 represents a lower alkyl group or a
substituted phenyl group); the other symbols have the same
definitions as those shown above.
The halogen atom represented by Z in formula (III) is
exemplified by fluorine, chlorine, bromine and iodine. The
lower alkyl group represented by R6 is exemplified by alkyl
groups having 1 to 6 carbon atoms, such as methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,
pentyl, isopentyl, neopentyl, tert-pentyl, l-ethylpropyl,
hexyl, isohexyl, l,l-dimethylbutyl, 2,2-dimethylbutyl, 3,3-
dimethylbutyl and 2-ethylbutyl, with preference given to
those having 1 to 4 carbon atoms, such as methyl, ethyl,
propyl, isopropyl, butyl and isobutyl.
CA 02248784 l998-09-ll
W O 97/49703 PCTIJP97/02157
38
Useful substituents for the substituted phenyl group
represented by R6 include, for example, lower alkyl groups
(same as those mentioned to exemplify the lower alkyl ~roup
represented by R6 above), lower alkoxy groups (e.g., those
having 1 to 4 carbon atoms, such as methoxy, ethoxy,
propoxy, isopropoxy and butoxy)~ halogen atoms (e.g.,
fluorine, chlorine, bromine, iodine), nitro groups, cyano
groups and carboxyl groups.
This method is conducted by reacting compound (III) or
a salt thereof with compound (IV) in the presence of a
base. The salt of compound (III) is exemplified by the
acid addition salts mentioned to exemplify acids that form
an acid addition salt with compound (I). This reaction is
normally carried out in a solvent; a solvent that does not
interfere with the reaction is chosen as appropriate. Such
solvents include, for example, alcohols such as methanol,
ethanol, propanol, isopropanol, butanol and tert-butanol;
ethers such as dioxane, tetrahydrofuran, diethyl ether,
tert-butyl methyl ether, diisopropyl ether and ethylene
glycol-dimethyl ether; esters such as ethyl formate, ethyl
acetate and n-butyl acetate; halogenated hydrocarbons such
as dichloromethane, chloroform, carbon tetrachloride,
trichlene and 1,2-dichloroethane; hydrocarbons such as n-
hexane, benzene and toluene; amides such as formamide, N,N-
dimethylformamide and N,N-dimethylacetamide; ketones such
as acetone, methyl ethyl ketone and methyl isobutyl ketone;
nitriles such as acetonitrile and propionitrile; dimethyl
sulfoxide, sulfolane, hexamethylphosphoramide and water;
these solvents are used as simple or mixed solvents.
Useful bases include, for example, Cl_6 alkyl or aryl
lithiums such as methyl lithium, ethyl lithium, n-butyl
lithium, sec-butyl lithium, tert-butyl lithium and phenyl
lithium; lithium alkylamides having 2 to 6 carbon atoms,
such as lithium dimethylamide, lithium diethylamide and
lithium diisopropylamide; metal hydrides such as lithium
CA 02248784 1998-09-11
- W O 97/49703 PCT1JP97/02157
39
hydride and sodium hydride; metal alkoxides having l to 6
carbon atoms, such as lithium ethoxide, lithium tert-
butoxide, sodium methoxide, sodium ethoxide and potassium
tert-butoxide; amides such as lithium amide, potassium
amide and sodium amide; inorganic bases such as lithium
hydroxide, potassium hydroxide, sodium hydroxide, sodium
carbonate, potassium carbonate and sodium hydrogen
carbonate; and tertiary amines such as triethylamine,
tri(n-propyl)amine, tri(n-butyl)amine,
diisopropylethylamine, cyclohexyldimethylamine, pyridine,
lutidine, y-collidine, N,N-dimethylaniline, N-
methylpiperidine, N-methylpyrrolidine and N-
methylmorpholine. The reaction is carried out using l to 5
mol, preferably l to 3 mol, of compound (IV) per mol of
compound (III). Reaction temperature is normally about -80
to 100~C, preferably -50 to 60~C. Reaction time is
normally 1 minute to 72 hours, preferably 15 minutes to 24
hours, depending on the kinds of compounds (III) and (IV),
the kind of solvent, reaction temperature etc.
CA 02248784 1998-09-11
- W097t49703 PCT/~7/02157
Method B
Rl R2 N~R3
A ~ CoOR7 N~R3
(VI)
oR8 ~ (I)
(V) /~
S
0 NH2CN ~ 4
/ Cyclization
(VII)
Rl R2 1l 3
A X ~/ CONHCN' 4
oR8
(VIII)
In the above formulas, R7 represents hydrogen or a
lower alkyl group; R8 represents hydrogen or a hydroxyl
group-protecting group; the other symbols have the same
definitions as those shown above.
The lower alkyl group represented by R7 is exemplified
by the same lower alkyl groups as those mentioned to
exemplify the lower alkyl group used for R6 in method A.
The hydroxyl group-protecting group represented by R8
may be any one, as long as it does not interfere with the
reaction; preferable examples thereof include ether-forming
protecting groups such as methoxymethyl, benzyloxymethyl,
tert-butoxymethyl, 2-methoxyethoxymethyl, 2-
(trimethylsilyl)ethoxymethyl, methylthiomethyl, 2-
tetrahydropyranyl, 4-methoxy-4-tetrahydropyranyl, 2-
tetrahydrofranyl, benzyl, p-methoxybenzyl, p-nitrobenzyl,
CA 02248784 1998-09-11
~ W097/49703 PCT/~97/02157
41
o-nitrobenzyl and trityl; silyl ether-forming protecting
groups such as trimethylsilyl, triethylsilyl,
- triisopropylsilyl, isopropyldimethylsilyl,
diethylisopropylsilyl, tert-butyldimethylsilyl, tert-
- 5 butyldiphenylsilyl, tribenzylsilyl, triphenylsilyl and
methyldiphenylsilyl; and ester-forming protecting groups
such as formyl, acetyl, chloroacetyl, dichloroacetyl,
trichloroacetyl, pivaloyl and benzoyl.
When R8 in formula (V) is hydrogen, compound (V) or a
salt thereof is reacted with compound (VI). The salt of
compound (V) is exemplified by acid adduct salts with the
acids mentioned to exemplify acids that form an acid adduct
salt with compound (I). This reaction is normally carried
out in a solvent and, if necessary, in the presence of a
base. Such solvents and bases are identical to the
solvents and bases mentioned for method A above. The
reaction is carried out using l to lO mol, preferably l to
5 mol, of compound (VI) per mol of compound (V) or salt
thereof. Reaction temperature is normally about -30 to
200~C, preferably -lO to 150~C. Reaction time is normally
l minute to 120 hours, preferably 15 minutes to 48 hours,
depending on the kinds of compounds (V) and (VI), the kinds
of solvent and base, reaction temperature etc.
Compound (I) can also be produced by producing
compound (VIII) from compounds (V) and (VII) and cyclizing
compound (VIII). This method involves the acylation of
compound (VII) or a salt thereof with compound (V), a salt
thereof or a reactive derivative thereof.
Specifically, free acid (V), a salt thereof (inorganic
salt, organic salt) or a reactive derivative thereof (e.g.,
acid halide, acid azide, acid anhydride, mixed acid
anhydride, active amide, active ester, active thioester
etc.) is subjected to acylation reaction. Inorganic salts
include alkali metal salts (e.g., sodium salt, potassium
CA 02248784 1998-09-11
- W O 97/49703 PCTtJP97/021S7
42
salt) and alkaline earth metal salts (e.g., calcium salt).
Organic salts include, for example, trimethylamine salt,
triethylamine salt, tert-butyldimethylamine salt,
dibenzylmethylamine salt, benzyldimethylamine salt, N,N-
dimethylaniline salt, pyridine salt and quinoline salt.
Acid halides include, for example, acid chloride and acid
bromide. Mixed acid anhydrides include mono-Cl_4
alkylcarbonic acid mixed acid anhydrides te.g., mixed acid
anhydrides of free acid (V) and monomethylcarbonic acid,
monoethylcarbonic acid, monoisopropylcarbonic acid,
monoisobutylcarbonic acid, mono-tert-butylcarbonic acid,
monobenzylcarbonic acid, mono(p-nitrobenzyl)carbonic acid,
monoallylcarbonic acid etc.), C1_6 aliphatic carboxylic
acid mixed acid anhydrides te.g., mixed acid anhydrides of
free acid (V) and acetic acid, cyanoacetic acid, propionic
acid, butyric acid, isobutyric acid, valeric acid,
isovaleric acid, pivalic acid, trifluoroacetic acid,
trichloroacetic acid, acetoacetic acid etc.), C7_ll
aromatic carboxylic acid mixed acid anhydrides (e.g., mixed
acid anhydrides of free acid (V) and benzoic acid, p-toluic
acid, p-chlorobenzoic acid etc.) and organic sulfonic acid
mixed acid anhydrides (e.g., mixed acid anhydrides with
methanesulfonic acid, ethanesulfonic acid, benzenesulfonic
acid, p-toluenesulfonic acid etc.). Active amides include
amides with nitrogen-containing heterocyclic compounds
[e.g., acid amides of free acid (V) and pyrazole,
imidazole, benzotriazole etc.; these nitrogen-containing
heterocyclic compounds may be substituted for by Cl-4
alkyls (e.g., methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, sec-butyl, tert-butyl), Cl_6 alkoxys (e.g.,
methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy),
halogen atoms (e.g., fluorine, chlorine, bromine), oxo,
thioxo, Cl_6 alkylthios ~e.g., methylthio, ethylthio,
propylthio, butylthio) etc.~.
CA 02248784 1998-09-11
W O 97/49703 PCT/JP971021S7- -
43
Active esters include, for example, organic phosphoric
acid esters (e.g., diethoxyphosphoric acid esters,
diphenoxyphosphoric acid esters), p-nitrophenyl ester, 2,4-
dinitrophenyl ester, cyanomethyl ester, pentachlorophenyl
ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide
ester, l-hydroxybenzotriazole ester, 6-chloro-1-
hydroxybenzotriazole ester and l-hydroxy-lH-2-pyridone
ester. Active thioesters include esters with aromatic
heterocyclic thiol compounds [their heterocyclic rings may
be substituted for by Cl_4 alkyls (e.g., methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl),
Cl_6 alkoxys (e.g., methoxy, ethoxy, propoxy, isopropoxy,
butoxy, tert-butoxy), halogen atoms (e.g., fluorine,
chlorine, bromine), Cl_6 alkylthios (e.g., methylthio,
ethylthio, propylthio, butylthio) etc.] [e.g., 2-
pyridylthiol ester, 2-benzothiazolylthiol ester].
The salt of compound (VIl) is exemplified by salts
with alkali metals (e.g., potassium, sodium, lithium),
salts with alkaline earth metals (e.g., calcium, magnesium)
and acid addition salts (acid adduct salts with the acids
mentioned to exemplify acids that form an acid addition
salt with compound (I)).
This reaction is normally carried in a solvent; a
solvent that does not interfere with the reaction is chosen
as appropriate. Such solvents include, for example, ethers
such as dioxane, tetrahydrofuran, diethyl ether, tert-butyl
methyl ether, diisopropyl ether and ethylene glycol-
dimethyl ether; esters such as ethyl formate, ethyl acetate
and butyl acetate; halogenated hydrocarbons such as
dichloromethane, chloroform, carbon tetrachloride,
trichlene and 1,2-dichloroethane; hydrocarbons such as n-
hexane, benzene and toluene; amides such as formamide, N,N-
dimethylformamide and N,N-dimethylacetamide; ketones such
as acetone, methyl ethyl ketone and methyl isobutyl ketone;
nitriles such as acetonitrile and propionitrile; dimethyl
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157- -
44
sulfoxide, sulfolane, hexamethylphosphoramide and water;
these solvents are used as simple or mixed solvents. The
amount of compound (VII) used is normally 1 to 10 mol,
preferably 1 to 5 mol, per mol of compound (V). The
reaction is normally carried out in the temperature range
from -80 to 200~C, preferably from -40 to 150~C, and most
preferably from -30 to 100~C. Reaction time is normally 1
minute to 72 hours, preferably 15 minutes to 2~ hours,
depending on the kinds of compounds ~V) and (VII), the kind
of solvent (also mixing ratio in the case of a mixed
solvent), reaction temperature etc. When compound (V) is
used as an acid halide, the reaction can be carried out in
the presence of a deoxidizer to remove the released
hydrogen halide from the reaction system. Such deoxidizers
include, for example, inorganic bases such as sodium
carbonate, potassium carbonate, calcium carbonate and
sodium hydrogen carbonate; tertiary amines such as
triethylamine, tripropylamine, tributylamine,
cyclohexyldimethylamine, pyridine, lutidine, y-collidine,
N,~-dimethylaniline, N-methylpiperidine, N-
methylpyrrolidine and N-methylmorpholine; and alkylene
oxides such as propylene oxide and epichlorohydrin.
Compound (VIII) can be then cyclized to yield compound
(I) after the hydroxyl group-protecting group R8 is removed
as necessary. Depending on the kind of protecting group,
this deprotection reaction can be carried out by a known
method chosen as appropriate. For example, deprotection
can be achieved with an acid (e.g., formic acid, acetic
acid, propionic acid, hydrochloric acid, sulfuric acid,
hydrobromic acid, phosphoric acid, methanesulfonic acid,
ethanesulfonic acid, p-toluenesulfonic acid) or by
catalytic reduction [Raney nickel, platinum, palladium,
rhodium, or the like, for example, used as a catalyst at
normal pressure or increased pressure (2 to 100 atm)} in
the case of ether-forming protecting groups, with one of
CA 02248784 1998-09-11
w097/49703 PCT/~97/02157
the above-mentioned acids or a Lewis acid (e.g., zinc
chloride, zinc bromide, aluminum chloride, titanium
chloride) or a fluoride (e.g., potassium fluoride, sodium
fluoride, tetraethylammonium fluoride, tetra-n-
butylammonium fluoride) in the case of silyl ether-forming
protecting groups, or with a base (e.g., potassium hydrogen
carbonate, sodium hydrogen carbonate, potassium carbonate,
sodium carbonate, lithium hydroxide, potassium hydroxide,
sodium hydroxide) in the case of ester-forming protecting
groups. The reaction is normally carried out in a solvent;
such solvents are exemplified by the solvents used for
method A.
In the case of ether-forming protecting groups or
silyl ether-forming protecting groups, the amount of acid
or Lewis acid used is normally 0.001 to 100 mol, preferably
0.01 to 50 mol, per mol of compound (V). Reaction
temperature is normally -50 to 150~C, preferably -20 to
100~C. Reaction time is normally 1 minute to 72 hours,
preferably 15 minutes to 48 hours.
In the case of ester-forming protecting groups, the
amount of base used is normally 0.01 to 50 mol, preferably
0.1 to 20 mol, per mol of compound (V). Reaction
temperature is normally -20 to 150~C, preferably -10 to
100~C. Reaction time is normally 1 minute to 72 hours,
preferably 15 minutes to 48 hours.
Compound (VIII) thus deprotected is cyclized to
compound (I). This reaction is normally carried out in a
solvent. Such solvents are exemplified by the solvents
used for method A. Reaction temperature is normally -10 to
200~C, preferably -5 to 150~C. In this reaction, a base
may be used as a catalyst; such bases are exemplified by
the bases used for method A. To promote the reaction,
there may be used, for example, 2-chloro-3-
methylbenzoxazolium tetrafluoroborate, 2-chloro-3-
ethylbenzoxazolium tetrafluoroborate, 2-chloro-3-
CA 02248784 1998-09-11
W097/4g703 PCT/~97/02157
46
methylbenzothiazolium tetrafluoroborate, 2-chloro-3-
ethylbenzothiazolium fluoroborate, 2-chloro-l-
methylpyridinium tetrafluoroborate and 2-chloro-l-
ethylpyridinium tetrafluoroborate. The amount of reaction
promoter used is normally l to lO mol, preferably l to 3
mol, per mol of compound (VIII). A base is also used when
a reaction promoter is used. Such bases are exemplified by
the bases used for method A. Reaction temperature is
normally -30 to 150~C, preferably -20 to 100~C. Reaction
time is normally l minute to 72 hours, preferably l5
minutes to 48 hours.
When one of R3 and R4 is an acyl group, an esterified
carboxyl group or a carbamoyl group that may have a
substituent, compound (I) can be produced by methods C, D
and E below.
Of the compounds of formula (I), compound (Ib), which
has an acyl group for R3 or R4, can be produced by method
C.
Method C
A ~ N (IX) A ~ N
~ ~ R4bSo3H (Ib) N' 4a
In these formulas, R4a represents an acyl group; R4b
represents a group resulting from removal of the carbonyl
group or sulfonyl group from an acyl group; the other
symbols have the same definitions as those shown above.
The acyl group represented by R4a means an acyl group
represented by R4; the "acyl group" in the "group resulting
from removal of the carbonyl group or sulfonyl group from
an acyl group" represented by R4b means an acyl group
repregented by R4.
CA 02248784 1998-09-11
WO 97/49703 PCT/JP97/02157
47
In this reaction, compound (Ia) or a salt thereof can
be acylated with compound (IX) or (X) or a reactive
derivative thereof to yield compound (Ib). The salt of
compound (Ia) is exemplified by the same acid adduct salts
as those mentioned to exemplify the salt of compound (I).
The reactive derivative of compound (IX) is exemplified by
the reactive derivatives mentioned for method B. The
reactive derivative of compound (X) is exemplified by
sulfonic acid halides (e.g., sulfonyl bromide, sulfonyl
chloride) and sulfonic anhydride; the reaction is carried
out by the method described for method B or a modification
thereof.
In the compounds of formula (I), compound (Ic), which
has an esterified carboxyl group for R4, can be produced by
method D.
Method D
Rl R2 o
QR4C X ~
(XI) A r N
(Ia) ~ I //
~ N,R3
(Ic) ~R4C
In these formulas, R4c represents an esterified
carboxyl group; Q represents a halogen atom; the other
symbols have the same definitions as those shown above.
R4c is any one of the carboxyl groups that may be
esterified, represented by R4, except the free carboxyl
group.
The halogen represented by Q is exemplified by
fluorine, chlorine, bromine and iodine. This reaction is
carried out by reacting compound (Ia) or a salt thereof and
compound (XI). The salt of compound (Ia) is exemplified by
the acid adduct salts mentioned to exemplify the acid
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157
48
adduct salt of compound (Ia) for reaction D above. This
reaction is normally carried out in a solvent; such
solvents are exemplified by the solvents used for method B.
In this reaction, a hydrogen halide is released. ~o remove
the hydrogen halide, the reaction can be carried out in the
preSence of an acid scavenger. Such acid scavenger
includes, for example, inorganic bases such as sodium
carbonate, potassium carbonate, calcium carbonate and
sodium hydrogen carbonate; tertiary amines such as
triethylamine, tripropylamine, tributylamine,
cyclohexyldimethylamine, pyridine, lutidine, y-collidine,
N,N-dimethylaniline, N-methylpiperidine, N-
methylpyrrolidine and N-methylmorpholine; and alkylene
oxides such as propylene oxide and epichlorohydrin.
The amount of compound (XI) used is normally 1 to 20
mol, preferably l to 10 mol, per mol of compound (Ia).
Reaction temperature is normally -30 to 120~C, preferably
-20 to 80~C. Reaction time is normally 1 minute to 72
hours, preferably 15 minutes to 48 hours.
Of the compounds of formula (I), compound (Id), which
has a carbamoyl group which may be substituted, can be
produced by method E.
Method E
~(Nx~RI)o A ~ N
(Ic) ~ o ~ 3
(Id) R4d
~
~ R NCO
(Ia)-~''~ (XIII)
In these formulas, R4d represents a carbamoyl group
which may be substituted; R9, R10 and Rll, whether
identical or not, represent hydrogen or one of the
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157-
49
substituents mentioned to exemplify the substituent for the
carbamoyl group represented by R4, which may be
substituted; the other symbols have the same definitions as
those shown above.
In this method, compound (Ic) or a salt thereof can be
reacted with compound (XII) to yield compound (Id). The
salt of compound (Ic) is exemplified by the same acid
addition salts as those mentioned to exemplify the salt of
compound (I). This reaction is normally carried out in a
solvent; such solvents are exemplified by the solvents used
for method A. The amount of compound (XII) used is
normally 1 to 100 mol, preferably 1 to 30 mol, per mol of
compound (Ic). Reaction temperature is normally -30 to
200~C, preferably -10 to 100~C. Reaction time is normally
1 minute to 72 hours, preferably 15 minutes to 48 hours.
Compound (Id) can also be produced by reacting
compound (Ia) with isocyanate derivative (XIII). The
reaction is normally carried out in a solvent. Said
solvent may be any one, as long as it does not interfere
with the reaction. For example, ethers such as dioxane,
tetrahydrofuran, diethyl ether, tert-butyl ether,
diisopropyl ether and ethylene glycol-dimethyl ether;
esters such as ethyl formate, ethyl acetate and n-butyl
acetate; halogenated hydrocarbons such as dichloromethane,
chloroform, carbon tetrachloride, trichlene and 1,2-
dichloroethane; hydrocarbons such as n-hexane, benzene and
toluene; amides such as formamide, N,N-dimethylformamide
and N,N-dimethylacetamide; ketones such as acetone, methyl
ethyl ketone and methyl isobutyl ketone; nitriles such as
acetonitrile and propionitrile; dimethyl sulfoxide,
sulfolane, hexamethylphosphoramide are used as simple or
mixed solvents.
The amount of compound (XIII) used is normally 1 to 30
mol, preferably 1 to 15 mol, per mol of compound (Ia).
Reaction temperature is normally -20 to 150~C, preferably
CA 02248784 1998-09-11
W 0 97/49703 PCT/JP97/02157 - -
-10 to 100~C. Reaction time is normally 1 minute to 72
hours, preferably 15 minutes to 48 hours.
Method F
O 12
R112 ~ NRl o
~N Rl500C~ ~ N ~ ~ N
3 N~ R4 ~ o ~ N~ R4
0 ~XIV) p(R16~3 (XvII)
(XVI)
R12
Hydrolysis ~ = o ~ L N~ 4 Heating
hydrogenolysis 1 2\ R
R R COOH
(XVIII)
In these formulas, R12 represents hydrogen, a lower
alkyl group, a cycloalkyl group, an aralkyl group or an
acyl group; R13 and Rl4, whether identical or not,
represent hydrogen or a lower alkyl group; Rl5 represents
hydro~en, a lower alkyl group or an aralkyl group; R16
represents a lower alkyl group or an aryl group; the other
symbols have the same definitions as those shown above.
The lower alkyl group represented by R12 in formula
(XIV) is exemplified by alkyl groups having 1 to 6 carbon
atoms, such as methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl,
neopentyl, tert-pentyl, l-ethylpropyl, hexyl, isohexyl,
l,l-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl and
2-ethylbutyl, with preference given to those having 1 to 4
carbon atoms, such as methyl, ethyl, propyl, isopropyl,
butyl and isobutyl. The cycloalkyl group represented by
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157
Rl2 is exemplified by cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl, with
preference given to cyclopropyl, cyclobutyl, cyclopentyl
and cyclohexyl. The aralkyl group represented by RL2 is
exemplified by benzyl, phenethyl and phenylpropyl.
The acyl group represented by Rl2 is exemplified by
aliphatic acyl groups such as alkanoyl groups, alkenoyl
groups, cycloalkanecarbonyl groups and alkanesulfonyl
groups; aromatic acyl groups such as aroyl groups,
arylalkanoyl groups, arylalkenoyl groups and arenesulfonyl
groups; heterocyclic aromatic acyl groups such as aromatic
heterocyclic carbonyl groups and aromatic heterocyclic
alkanoyl groups; and non-aromatic heterocyclic carbonyl
groups (aliphatic heterocyclic carbonyl groups).
"Alkanoyl groups" mean alkylcarbonyl groups,
preferable examples thereof including lower alkanoyl groups
having l to 8 carbon atoms, such as formyl, acetyl,
propionyl, butyryl, isobutyryl, valeryl, isovaleryl,
pivaloyl and hexanoyl.
"Alkenoyl groups" mean alkenylcarbonyl groups,
preferable examples thereof including C3-6 alkenoyl groups
such as acryloyl, methacryloyl, crotonoyl and isocrotonoyl.
"Cycloalkanecarbonyl groups" mean cycloalkylcarbonyl
groups, preferable examples thereof including those having
4 to 7 carbon atoms, such as cyclopropanecarbonyl groups,
cyclobutanecarbonyl groups, cyclopentanecarbonyl groups and
cyclohexanecarbonyl groups.
"Alkanesulfonyl groups" mean alkylsulfonyl groups,
preferable examples thereof including those having l to 4
carbon atoms, such as mesyl, ethanesulfonyl and
propanesulfonyl.
"Aroyl groups" mean arylcarbonyl groups, preferable
examples thereof including those having 7 to ll carbon
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157- -
52
atoms, such as benzoyl, p-toluoyl, l-naphthoyl and 2-
naphthoyl.
"Arylalkanoyl groups" mean alkylcarbonyl groups
substituted for by an aryl group, preferable examples
thereof including C6_g aryl-C2_s alkanoyl groups such as
phenylacetyl, phenylpropionyl, hydroatropoyl and
phenylbutyryl.
"Arylalkenoyl groups" mean alkenylcarbonyl groups
substituted for by an aryl group, preferable examples
thereof including C6_g aryl-C3_5 alkenoyl groups such as
cinnamoyl and atropoyl.
"Arenesulfonyl groups" mean arylsulfonyl groups,
preferable examples thereof including those having 6 to 8
carbon atoms, such as benzenesulfonyl and p-
toluenesulfonyl.
Preferable examples of "aromatic heterocyclic carbonyl
groups" include furoyl, thenoyl, nicotinoyl, isonicotinoyl,
pyrrolecarbonyl, oxazolecarbonyl, thiazolecarbonyl,
imidazolecarbonyl and pyrazolecarbonyl.
"Aromatic heterocyclic alkanoyl groups" mean
alkylcarbonyl groups substituted for by an aromatic
heterocyclic group, preferable examples thereof including
aromatic heterocyclic ring-C2_5 alkanoyl groups such as
thienylacetyl, thienylpropanoyl, furylacetyl,
thiazolylacetyl, l,2,4-thiadiazolylacetyl and
pyridylacetyl.
Preferable examples of "non-aromatic heterocyclic
carbonyl groups" include aliphatic heterocyclic carbonyls
such as azetidinylcarbonyl, pyrrolidinylcarbonyl and
piperidinylcarbonyl.
The lower alkyl group represented by Rl3, Rl4, Rl5 or
R16 in formulas (XIV), (XV~ and (XVI) is exemplified by
lower alkyl groups represented by Rl2. The aralkyl group
represented by Rl5 is exemplified by aralkyl groups
CA 0224X784 1998-09-11
W097/49703 PCT/~97/02157
represented by Rl2. The aryl group represented by R16 is
exemplified by phenyl, naphthyl, anthryl, phenanthryl and
acenaphthylenyl groups, with preference given to phenyl and
naphthyl. These aryl groups may have 1 to 5 substituents.
Such substituents include alkyl groups having 1 to 3 carbon
atoms (e.g., methyl, ethyl, propyl), alkoxy groups having 1
to 3 carbon atoms (e.g., methoxy, ethoxy, propoxy) and
halogen atoms (e.g., fluorine, chlorine, bromine, iodine).
In this method, compounds (XIV) and (XV) are reacted
in the presence of compound (XVI) to yield compound (XVII).
This reaction is normally carried out in a solvent; a
solvent that does not interfere with the reaction is chosen
as appropriate. Such solvents include, for example,
alcohols such as methanol, ethanol, propanol, isopropanol,
butanol and tert-butanol; ethers such as dioxane,
tetrahydrofuran, diethyl ether, tert-butyl methyl ether,
diisopropyl ether and ethylene glycol-dimethyl ether;
esters such as ethyl formate, ethyl acetate and n-butyl
acetate; halogenated hydrocarbons such as dichloromethane,
chloroform, carbon tetrachloride, trichlene and 1,2-
dichloroethane; hydrocarbons such as n-hexane, benzene and
toluene; amides such as formamide, N,N-dimethylformamide
and N,N-dimethylacetamide; ketones such as acetone, methyl
ethyl ketone and methyl isobutyl ketone; nitriles such as
acetonitrile and propionitrile; dimethyl sulfoxide,
sulfolane, hexamethylphosphoramide and water; these
solvents are used as simple or mixed solvents.
Reaction temperature is normally -80 to 150~C,
preferably -50 to 120~C. The amount of each of compounds
(XV) and (XVI) used is normally 1 to 5 mol, preferably 1 to
3 mol, per mol of compound (XIV).
The ester of compound (XVII) is then subjected to
hydrolysis, hydrogenolysis, or the like, to yield compound
(XVIII).
CA 02248784 l998-09-ll
- W 097/49703 PCT/JP97/02157 - -
54
This hydrogenolysis reaction is normally carried out
in a solvent; a solvent that does not interfere with the
reaction is chosen as appropriate. Such solvents include,
for example, alcohols such as methanol, ethanol, propanol,
isopropanol, butanol and tert-butanol; ethers such as
dioxane, tetrahydrofuran, diethyl ether, tert-butyl methyl
ether, diisopropyl ether and ethylene glycol-dimethyl
ether; halogenated hydrocarbons such as dichloromethane,
chloroform, carbon tetrachloride, trichlene and 1,2-
dichloroethane; hydrocarbons such as n-hexane, benzene and
toluene; amides such as formamide, N,N-dimethylformamide
and N,N-dimethylacetamide; ketones such as acetone, methyl
ethyl ketone and methyl isobutyl ketone; nitriles such as
acetonitrile and propionitrile; dimethyl sulfoxide,
- 15 sulfolane, hexamethylphosphoramide and water; these
solvents are used as simple or mixed solvents.
This reaction is carried out in the presence of a
base. Preferably used bases include metal hydroxides such
as lithium hydroxide, potassium hydroxide, sodium hydroxide
and barium hydroxide, and metal carbonates such as
potassium carbonate, sodium carbonate and barium carbonate.
The amount of base used is normally 1 to 30 mol, preferably
1 to 10 mol, per mol of compound (XVII). Reaction
temperature is normally -30 to 150~C, preferably -10 to
120~C. Reaction time is normally 15 minutes to 48 hours,
preferably 30 minutes to 24 hours.
When compound (XVIII) is produced by a hydrogenolysis
reaction, the reaction is normally carried out using a
catalyst. This catalyst is preferably one for catalytic
reduction reaction, exemplified by platinum catalysts
(e.g., platinum oxide, platinum black, platinum-carbon),
palladium catalysts (e.g., palladium chloride, palladium-
carbon, palladium-calcium carbonate, palladium-barium
sulfate), rhodium catalysts (e.g., rhodium-carbon, rhodium-
alumina) and ruthenium catalysts (e.g., ruthenium oxide,
CA 02248784 1998-09-11
- W O 97/49703 PCT/JP97/02157
ruthenium-carbon), with greater preference given to
palladium catalysts. The reaction is normally carried out
in a solvent; a solvent that does not interfere with the
reaction is chosen as appropriate. Such solvents include,
for example, alcohols such as methanol, ethanol, propanol,
isopropanol, butanol and tert-butanol; ethers such as
dioxane, tetrahydrofuran, diethyl ether, tert-butyl methyl
ether, diisopropyl ether and ethylene glycol-dimethyl
ether; esters such as ethyl formate, ethyl acetate and n-
butyl acetate; hydrocarbons such as n-hexane, benzene and
toluene; amides such as formamide, N,N-dimethylformamide
and N-,N-dimethylacetamide; and water; these solvents are
used as simple or mixed solvents.
Reaction temperature is normally -10 to 120~C,
preferably 0 to 100~C. Although this reaction is normally
carried out at normal pressure, it may be carried out at
increased pressure in some cases. Such pressure is
preferably 1 to 200 atm.
Compound (XVIII) can be decarbonized by heating to
yield compound (I). This reaction is normally carried out
in a solvent; a solvent that does not interfere with the
reaction is chosen as appropriate. Such solvents include,
for example, alcohols such as methanol, ethanol, propanol,
isopropanol, butanol and tert-butanol; ethers such as
dioxane, tetrahydrofuran, diethyl ether, tert-butyl methyl
ether, diisopropyl ether and ethylene glycol-dimethyl
ether; esters such as ethyl formate, ethyl acetate and n-
butyl acetate; halogenated hydrocarbons such as
dichloromethane, chloroform, carbon tetrachloride,
trichlene and 1,2-dichloroethane; hydrocarbons such as n-
hexane, benzene and toluene; amides such as formamide, N,N-
dimethylformamide and N,N-dimethylacetamide; ketones such
as acetone, methyl ethyl ketone and methyl isobutyl ketone;
nitriles such as acetonitrile and propionitrile; dimethyl
CA 02248784 1998-09-11
- W097/4g703 PCT/~97tO2157
56
sulfoxide, sulfolane, hexamethylphosphoramide and water;
these solvents are used as simple or mixed solvents.
Reaction temperature is normally O to 180~C,
preferably 10 to 150~C. Reaction time is normally 5
minutes to 24 hours, preferably 10 minutes to 12 hours.
When a compound involved in each reaction described
above has an amino group, a carboxyl group or a hydroxyl
group as a substituent, the group may incorporate a
protecting group in common use in peptide chemistry and
other fields; the desired compound can be obtained by
removing the protecting group as necessary after reaction.
Useful amino group-protecting groups include, for
example, formyl group, Cl_6 alkylcarbonyl groups (e.g.,
acetyl, ethylcarbonyl), benzyl group, tert-butyloxycarbonyl
group, benzyloxycarbonyl group, 9-
fluorenylmethyloxycarbonyl group, allyloxycarbonyl group,
phenylcarbonyl group, Cl_6 alkyloxycarbonyl groups (e.g.,
methoxycarbonyl, ethoxycarbonyl), C7-lo aralkylcarbonyl
groups (e.g., benzylcarbonyl), trityl group, phthaloyl
group and N,N-dimethylaminomethylene group. These groups
may be substituted by 1 to 3 halogen atoms (e.g., fluorine,
chlorine, bromine), nitro group etc.
Useful carboxyl group-protecting groups include Cl_6
alkyl groups (e.g., methyl, ethyl, propyl, isopropyl,
butyl, tert-butyl), phenyl group, silyl group, benzyl group
and allyl group. These groups may be substituted for by 1
to 3 halogen atoms (e.g., fluorine, chlorine, bromine),
nitro group etc.
Useful hydroxyl group-protecting groups include
methoxymethyl group, allyl group, tert-butyl group, C7_l0
aralkyl groups (e.g., benzyl), formyl group, Cl_6
alkylcarbonyl groups (e.g., acetyl, ethylcarbonyl), benzoyl
group, C7_l0 aralkylcarbonyl groups te.g., benzylcarbonyl),
pyranyl group, furanyl group and trialkylsilyl groups.
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157
57
These groups may be substituted by 1 to 3 halogen atoms
(e.g., fluorine, chlorine, bromine), Cl 6 alkyl groups
(e.g., methyl, ethyl, propyl, isopropyl, butyl, tert-
butyl), phenyl group, C7-lo aralkyl groups (e.g., benzyl),
nitro group etc.
These protecting groups can be removed by commonly
known methods or modifications thereof, including those
using acids, bases, reduction, ultraviolet rays, hydrazine,
phenylhydrazine, sodium N-methyldithiocarbamate,
tetrabutylammonium fluoride, palladium acetate etc.
When a compound is obtained in a free form by each of
the above-described reactions of the present invention, it
may be converted to a salt by a conventional method; when
it is obtained as a salt, it may be converted to a free
form or another salt by a conventional method.
Compound (I) thus obtained can be isolated and
purified from the reaction mixture by commonly known means
such as extraction, concentration, neutralization,
filtration, recrystallization, column chromatography and
thin-layer chromatography.
A salt of compound (I) can be produced by, for
example, adding one of the above-described inorganic acids
or organic acids to compound (I) by a commonly known means.
Compounds (III) and (IV), used as starting compounds
in method A above, can be produced by, for example, the
method described in US Patent No. 4,584,385 or a method
based thereon.
Compound (V), used as a starting compound in method B
above, can be produced by, for example, the method
described in the Journal of Medicinal Chemistry, 21, 82
(1978) or Chemistry Letters, 166 (1980) or a method based
thereon; compound (VI) can be produced by, for example, the
method described in the Journal of Organic Chemistry, 42,
3608 (1977) or a method based thereon; compound (VII) can
CA 02248784 1998-09-11
W097/49703 PC~/~97/02157
58
be produced by the method described in the Journal of the
Chemical Society, 9S, 132 (1909) or a method based thereon.
In addition to the above-mentioned processes, compound
(I) can also be produced by the method described in US
Patent No. 4,584,385 or a method based thereon.
Although compound (I) can be produced by chemical
processes as described above, it can also be produced using
microorganisms. Of the compounds of formula (I),
indolmycin, can be produced by, for example, the methods
described in the literature [K.V. Rao, Antibiotics and
Chemotherapy (Washington, D.C.), 10, 312 (1960); W.S. Marsh
et al., ibid., 10, 316 (1960); Schach von Wittenau, M. et
al., J. Am. Chem. Soc. 83, 4678 (1961), ibid., 85, 3425
(1963)], using as producer strains Streptomycesgriseus subsp.
griseus ATCC 1264 (American Type Culture Collection
Catalogue of Bacteria & Bacteriophages, 18th edition, 1992)
etc. Streptomyces sp. HC-21, a new strain, can also be used
as a producer strain.
The microorganism used for the method of indolmycin
production of the present invention is the Streptomyces sp.
HC-21 strain (hereinafter also referred to as "HC-21
strain") isolated from a soil sample from Tenninkyo,
Asahikawa-shi, Hokkaido, Japan.
According to the method described in the International
Journal of Systematic Bacteriology, 16(3), 313-340 (1960),
the HC-21 strain is characterized as follows: All findings
on medium were obtained during 14 days of cultivation and
observation at 28~C, unless otherwise stated.
(I) Morphological characteristics
The aerial mycelia elongate in simple branches from
well elongated and branched substrate mycelia, with gently
waved or key-shaped spore chains (normally 10 to 50 spores
or more) on their tips. No whirls are noted. Spores are
CA 02248784 l998-09-ll
W O 97/49703 PCT/JP97/02157'
cylindrical (1.1 to 1.2 x 1.4 to l.S ~m) and have a spiny
surface.
~II) Nature in culture
Degree of growth (G), growth and color tone of aerial
mycelia (AM), back face color tone (R), presence or absence
and color tone of soluble pigment (SP) etc. on various
media are described below. For the description of color,
standard color tone symbols in parentheses are based on the
Color ~armony Manual of Container Corporation of America,
4th edition, 1958.
CA 02248784 1998-09-11
- W O 97/49703 PCT/JP97/02157- -
Table 1
(a) Sucrose-nitrate- G : Poor, light ivory (2ca)
agar medium AM: None
R : Light ivory (2ca)
SP: None
(b) Glucose-aspara- G : Good, ivory (2ea)
gine-agar medium AM: Good, ivory (2ea)
R : Light yellowish brown (2ia) to yellowish
brown (3na)
SP: None
(c) Glycerol-aspara- G : Good, yellowish brown (3ic)
gine-agar medium AM: Good, Light yellowish gray (2gc)
R : Yellowish brown (3La) to brown (41a)
SP: None
(d) Starch-inorganic G : Moderate, ivory (2ec)
salt-agar medium AM: Moderate, light greenish gray (2cb)
R : Light yellowish brown (2ga) to light
yellowish qray (2gc)
sP: None
(e) Tyrosine-agar me- G : Good, light yellowish brown (2ga)
dium AM: Ivory (2ea)
R : Light yellowish brown (21a) to yellowish
brown (31c)
SP: None
(~) Enriched agar me- G : Moderate, light grayish reddish brown (5ic)
dium AM: Poor, white
R : Light grayish brown (4ga) to reddish brown
(61a)
SP: None
(g) Yeast extract-malt G : Good, yellowish brown (4ia)
extract-agar me- AM: Moderate, light grayish brown (5ga) to
dium grayish yellowish brown (4ge)
R : Yellowish brown (3ia) to reddish yellowish
brown (Spa)
SP: None
(h) Oatmeal-agar me- G : Good, light grayish brown (4gc)
dium AM: Moderate, light reddish white (5ea) to
grayish brown (5gc)
R : Bright reddish brown (6ia) to grayish brown
(5gc)
SP: None
(i) Peptone-yeast G : Moderate, ivory (2ea), localized
extract-iron-agar AM: None
medium R : Light yellowish brown (2ga) to yellowish
grayish brown (3ia)
SP: None
(III) Physiological nature
(a) Growth temperature range : 11 to 29~C
optimal growth temperature range: 18 to 24~C
(b) Nitrate reduction : Weakly positive
(c) Gelatin liquefaction : Negative
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97102157
61
(glucose-peptone-gelatin medium)
(d) Starch hydrolysis : Negative
(e) Defatted milk coagulation : Negative
Defatted milk peptonization : Negative
(f) Melanin-like pigment formation
Tyrosine-agar medium : Negative
Peptone-yeast extract-
iron-agar medium : Negative
(g) Carbon source assimilation (agar medium
containing pridham and gottlieb)
L-arabinose: -
D-xylose : -
D-glucose : +~
D-fructose : +
Sucrose : -
Inositol : -
L-rhamnose : ++
Raffinose : -
D-mannitol : -
Control : -
(Note) ++: relatively good growth
+ : growth noted
+ : + or - indeterminable
- : no growth
(IV) Cell analysis
Analysis in accordance with the method of Hasegawa et
al. [Journal of General Applied Microbiology 29, 319-322
(1983)] identified the diaminopimelic acid in the
hydrochloric acid hydrolyzate of cells as the LL-
configuration.
Judging from the results shown above, specifically the
light yellowish brown to grayish brown aerial mycelia,
gently waved or key-shaped spore chains, spiny spore
3~ surfaces, diaminopimelic acid in the LL-configuration, and
other findings, it is evident that this strain belongs to
CA 02248784 1998-09-ll
W 097149703 PCTIJP97/02157 - -
62
the genus Streptomyces; the strain was designated Streptomyces
sp. HC-21.
The Streptomyces sp. HC-21 strain as such is
characterized by the capability of L-rhamnose assimilation
and spiny spore surfaces.
The S~reptomyces sp. HC-21 strain as such has been
deposited under accession number IFO-15984 at the Institute
for Fermentation, Osaka (foundation), since June 12, 1996,
and under accession number FERM BP-5571 at the National
Institute of Bioscience and Human-Technology, Agency of
Industrial Science and Technology, Ministry of
International Trade and Industry of the Japan (1-3, Higashi
l-chome, Yatabe, Tsukuba City, Ibaraki Prefecture), since
June 25, 1996.
The bacteria of the genus Streptomyces can undergo
variation, naturally or by mutagens, as a general nature of
microorganisms. Even the various variants obtained by, for
example, irradiation with radiations such as X rays, gamma
rays and ultraviolet rays, single spore separation,
treatment with various chemicals, cultivation on drug-
containing media, and other means, or naturally-occurring
mutants, are all usable for the method of the present
- invention, as long as they are capable of producing
indolmycin.
Although the culture medium for the method of the
present invention may be liquid or solid, as long as it
contains nutrient sources usable by the strain used, a
liquid medium is preferred for large-scale treatment. The
medium is supplemented as appropriate with assimilable
nutrient sources, digestible nitrogen sources, inorganic
substances, and trace nutrients.
Carbon sources include, for example, glucose, lactose,
sucrose, maltose, dextrin, starch, glycerol, mannitol,
sorbitol, oils and fats (e.g., soybean oil, olive oil, rice
bran oil, sesame oil, lard oil, chicken oil); nitrogen
CA 02248784 1998-09-11
- W097/49703 PCT/~97/02157
sources include, for example, meat extract, yeast extract,
dry yeasts, soybean flour, corn steep liquor, peptone,
cottonseed flour, blackstrap molasses, urea, ammonium salts
(e.g., ammonium sulfate, ammonium chloride, ammonium
nitrate, ammonium acetate) and others. Also used as
appropriate are salts containing sodium, potassium,
calcium, magnesium etc., metal salts such as those of iron,
manganese, zinc, cobalt, nickel etc., salts of phosphoric
acid, boric acid etc., and salts of organic acids such as
acetic acid and propionic acid. Additionally, amino acids
(e.g., glutamic acid, aspartic acid, alanine, lysine,
valine, methionine, proline), vitamins (e.g., Bl, B2,
nicotinic acid, B12, C), nucleic acids (e.g. purine,
pyrimidine and derivatives thereof) etc. may be contained.
It is of course common practice to add inorganic or organic
acids, alkalis, buffers etc. for regulation of the medium's
pH, and appropriate amounts of oils and fats, surfactants
etc. for defoaming.
Cultivation may be achieved by standing culture,
shaking culture, spinner culture, or the like. For large-
scale treatment, submerged spinner culture is of course
desirable.
Although culturing conditions vary depending on the
condition and composition of the medium, the kind of
strain, and the means of cultivation, it is normally
recommended that temperature and initial pH be 15 to 26~C
and about 5 to 9, respectively. It is desirable that
temperature in the middle stage of cultivation and initial
pH be 20 to 25~C and about 6 to 8, respectively. Duration
of cultivation also varies depending on the above-mentioned
conditions but it is recommended that cultivation be
continued until the concentration of the desired bioactive
substance reaches maximum. It normally takes about l to lO
days in the case of shaking culture or spinner culture
using a liquid medium.
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157
64
The resulting bioactive substance indolmycin can be
extracted and purified from the culture on the basis of its
chemical nature.
Because indolmycin is produced in the culture broth
and cells, it can be purified by separating the culture
broth and cells by filtration or centrifugation from the
culture, extracting it from the resulting filtrate or
centrifugal supernatant using an organic solvent, or
extracting it from cells using an organic solvent, and
isolating it from each extract or the combined extract.
For industrial purposes, it is advantageous to purify
indolmycin from the extract obtained by adding an organic
solvent such as methanol, acetone, butanol or ethyl acetate
directly to the culture, with the cell separation operation
omitted.
Because indolmycin is a weakly basic oil-soluble
substance, its collection from the culture broth permits
the use of means of separation and purification in common
use for collection of related microbial metabolites. For
example, methods based on solubility differences from
impurity substances and chromatographies using various
carriers such as activated charcoal nonionic high porous
resin, silica gel, alumina and dextran gel can be used
singly or in combination.
The method of isolating and collecting indolmycin from
the culture is hereinafter described specifically. First,
cells are removed by filtration from the culture broth; the
resulting supernatant is adjusted to appropriate pH; a
solvent such as ethyl acetate is added, followed by
vigorous stirring, to yield an ethyl acetate layer. The
organic layer obtained is sequentially washed with alkali,
acid and water, after which it is concentrated; the
resulting concentrate is subjected to silica gel column
chromatography. Useful developing solvents include, for
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157 - -
example, chloroform-methanol or hexane-acetone mixed
sol~ents. After the effective fractions are combined and
concentrated, the concentrate is subjected to Sephadex LH-
20 chromatography. Useful developing solvents are methanol
and mixed solvents such as hexane-toluene-methanol and
hexane-methylene chloride-methanol. After concentration,
the eluate containing the effective fractions is purified
by preparative high performance liquid chromatography. The
column packing used here is ODS-SH343 S-15 (produced by
Yamamura Kagaku Kenkyujo); the solvent system used is a
combination of 0.02 M phosphate buffer (pH 6.3) and 26%
CH3CN.
Best Mode for Carring Out the Invention
The present invention is hereinafter described in more
detail by means of, but not limited to, the following
working examples, experimental examples and preparation
example. In the description below, "room temperature"
means about 15 to 30~C.
[Examples]
Example 1
A platinum loopful of Streptomyces sp. HC-21 strain,
previously sufficiently grown on a slant medium consisting
of yeast extract-maltose extract-agar, was inoculated to a
sterile 2 1 Sakaguchi flask containing 500 ml of a seed
medium of pH 7.0 consisting of 2% glucose, 3% soluble
starch, 1% corn steep liquor, 1% fresh soybean flour, 0.5%
polypeptone, 0.3% sodium chloride and 0.5% precipitating
calcium carbonate, and cultured at 24~C on a reciprocal
shaker for 2 days. To this 500 ml culture broth was
injected 120 1 of a principal medium of pH 7.0 consisting
of 2.0% glucose, 3.0% soluble starch, 1.0% defatted soybean
flour, 0.3% corn steep liquor, 0.1% yeast extract, 0.5%
polypeptone, 0.2% oatmeal agar, 0.3% sodium chloride, 0.5%
precipitating calcium carbonate, 0.05% ACTCOL 31-56
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157
(produced by Takeda Chemical Industries, Ltd., Japan) and
0.05% silicone, followed by transplantation to a sterile
200 1 fermentation jar and 90 hours of cultivation under
conditions of 24~C temperature, 1.1 kg/cm2 internal
pressure, 120 l/min aeration rate and 120 rpm stirring
rate.
The 120 1 of the culture broth thus obtained was
filtered using Hyflo Super Cel to yield 110 1 of a
filtrate. This filtrate was adjusted to pH 3.0 with dilute
sulfuric acid; an equal amount of ethyl acetate was added,
followed by vigorous stirring, to yield 80 1 of an ethyl
acetate layer. This ethyl acetate layer was washed with 30
1 of a 2% NaHCO3 solution, then with 30 1 of a 0.02 N HCl
-15 solution, and thoroughly washed with water, after which it
was concentrated under reduced pressure to yield about 30 g
of a concentrate. This concentrate was passed through a
silica gel column (0.8 1) to adsorb the active ingredient,
followed by sequential elution with 4 1 of hexane-acetone
(80:20), 4 1 of hexane-acetone (50:50) and 4 1 of hexane-
acetone (20:80). The effective fractions were combined and
concentrated under reduced pressure to yield 1.53 g of a
concentrate. This concentrate was dissolved in methanol;
the resulting solution was passed through a column (2 1) of
Sephadex LH-20 (produced by Pharmacia, Sweden), thoroughly
washed previously; the effective eluted fractions from 1.3
1 to 1.7 1 were combined and concentrated under reduced
pressure to yield 490 mg of a powder. This powder was
further developed (20 ml/min, 20 ml fractions) with a
solvent system of 0.02 M phosphate buffer (pH 6.3) and 26%
CH3CN, using preparative liquid chromatography (Hitachi
model L-6250, detector L-4000, YMC-Pack, ODS SH343 S-15 120
A, 214 nm), to yield effective fractions (fraction Nos. 30
through 39). After the CH3CN was removed, the effective
fractions were washed with water and again extracted with
ethyl acetate; the ethyl acetate layer was concentrated
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157
under reduced pressure to yield 315 mg of crystalline
indolmycin.
Elemental analysis (for Cl4Hl5N3O2):
Calculated: C, 65.35; H, 5.88; N, 16.33
Found : C, 65.14; H, 5.87; N, 16.07
The physicochemical properties also agreed well with
those of indolmycin.
Example 2
(5S)-2-(N-benzyloxycarbonyl-N-methyl)amino-5[(lR)-l-(indol-
3-yl)ethyl]-2-oxazolin-4-one
A solution of indolmycin (100 mg) in tetrahydrofuran
(5 ml) was cooled to -30~C. To this cooled solution,
triethylamine (0.217 ml) and carbobenzoxychloride (0.167
ml) was subsequently added dropwise at -30~C. The whole
was allowed to warm up to 0~C and was stirred for 80
minutes. Ethyl acetate was added to the reaction mixture.
The mixture was washed with water, saturated aqueous sodium
hydrogen carbonate solution and brine respectively and then
the ethyl acetate solution was dried over MgSO4. Removal
of the organic solvent gave a residue, which was
recrystalized with isopropylether to afford the titled
compound (115 mg, 75.5~). m.p. 136-138~C.
IR (KBr) cm~l: 3299, 1748.
lH-NMR (CDC13) ~: 1.39(3H,d,J=7.2Hz), 3.34(3H,s), 3.81-
3.93(1H,m), 5.10(1H,d,J=2.8Hz), 5.31(2H,s), 6.97-
7.40(8H,m), 7.62(1H,d,J=7.4Hz), 7.95(1H,bs).
Example 3
Following the same procedure as described in the
Example 2, the following compounds were prepared.
(5S)-2-(N-ethoxycarbonyl-N-methyl)amino-5[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3293, 1769, 1738.
lH-NMR (CDC13) ~: 1.34(3H,d,J=7.0Hz), 1.41(3H,t,J=7.2Hz),
3.32(3H,s), 3.88(1H,m), 4.35(2H,q,J=7.2Hz),
CA 02248784 1998-09-11
W O 97/49703 PCT/JP971021~7
68
5.10(1H,d,J=2.6Hz), 7.07-7.26(3H,m), 7.36(1H,d,J=8.2Hz),
7.67(1H,d,J=8.2Hz), 8.15(1H,bs).
(SS)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-
isopropoxycarbonyl-N-methyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3272, 1732.
H-NMR (CDC13) ~: 1.36(6H,d,J=6.4Hz), 1.43(3H,d,J=7.4Hz),
3.31(3H,s), 3.91(1H,m), 5.08(1H,m), 5.12(1H,d,J=2.6Hz),
7.07-7.35(3H,m), 7.37(lH,d,J=8.OHz), 7.67(lH,d,J=7.8Hz),
8.17(lH,bs).
(5S)-2-[N-(2-ethylhexyloxycarbonyl)-N-methyl]amino-5-
[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3298, 1769, 1742.
H-NMR (CDC13) ~: 0.90(6H,t,J=7.6Hz), 1.26-1.60(9H,m),
3.29(3H,s), 3.90(1H,m), 4.19(2H,d,J=5.8Hz),
15 5.12(1H,d,J=3.0Hz), 7.06-7.38(4H,m), 7.66(1H,d,J=7.6Hz),
8.14(1H,bs).
(5S)-2-[N-(4-acetoxybenzyloxycarbonyl)-N-methyl]amino-
5-~(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3333, 1746.
20 lH-NMR (CDC13) ~; 1.24(3H,d,J=7.4Hz), 2.41(3H,s),
3.50(3H,s), 3.80(1H,m), 5.09(1H,d,J=2.8Hz), 5.30(2H,s),
6.52(1H,d,J=2.2Hz), 7.03-7.46(7H,m), 7.57(1H,d,J=7.6Hz),
8.56(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(4-
nitrobenzyloxycarbonyl)-N-methyl]amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3299, 1773, 1746.
H-NMR (CDC13) ~: 1.44(3H,d,J=7.2Hz), 3.35(3H,s),
3.90(1H,m), 5.15(1H,d,J=2.6Hz), 5.37(2H,s), 7.03-
7.24(3H,m), 7.37(1H,d,J=8.2Hz), 7.47(2H,d,J=8.8Hz),
7.64(1H,d,J=8.4Hz), 8.10(1H,bs), 8.14(2H,d,J=8.8Hz).
(5S)-5-[(lR)-l-(indol-3-yl)ethylJ-2-(N-methyl-N-
phenoxycarbonyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3281, 1779, 1746.
lH-NMR (CDC13) ~: 1.45(3H,d,J=7.2Hz), 3.47(3H,s),
3.91(1H,m), 5.15(1H,d,J=3.0Hz), 7.03-7.49(9H,m),
7.67(1H,d,J=7.4Hz), 8.05(1H,bs).
CA 02248784 1998-09-11
~ W097/49703 PCTl~97/02157
69
Example 4
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(N-(4-
methoxyphenyl)carbamoyl)-N-methyl]amino-2-oxazolin-4-one
To a mixture of indolmycin (150 m~) and
dichloromethane (3 ml) was added 4-methoxyphenylisocyanate
(261 mg) under ice cooling. After the mixture was sitrred
for 2 hours at room temperature, the solvent was distilled
off to give a residue. After isopropylether was added to
the residue, the titled compound was obtained as a crystal
(213 m~, 89.9%).
IR (KBr) cm~l: 3382, 1717.
H-NMR (CDC13) ~: 1.48(3H,d,J=7.0Hz), 3.34(3H,s),
3.78(3H,s), 3.95(1H,m), 5.08(1H,d,J=3.0Hz),
6.83(2H,d,J=9.2Hz), 7.11-7.40(6H,m), 7.67(1H,d,J=8.4Hz),
8.21(1H,bs), 11.21(1H,bs).
Example 5
Following the same procedure as described in the
Example 4, the following compounds were prepared.
(5S)-2-[N-(N-(4-chlorophenyl)carbamoyl)-N-
methyl]amino-5-[(lR)-2-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3372, 1713.
lH-NMR (CDC13) ~: 1.49(3H,d,J=7.4Hz), 3.34(3H,s),
3.97(lH,m), 5.09(lH,d,J=3.2Hz), 7.15-7.47(8H,m),
7.67(1H,d,J=7.4Hz), 8.17(1H,bs), 11.48(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl~-2-[N-(N-(2-
phenylethyl)carbamoyl)-N-methyl]amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3243, 1713.
lH-NMR (CDC13) ~: 1.44(3H,d,J=7.4Hz), 2.82(2H,t,J=8.0Hz),
3.28(3H,s), 3.47(2H,m), 3.91(1H,m), 5.02(1H,d,J=3.0Hz),
7.11-7.40(9H,m), 7.65(1H,d,J=7.8Hz), 8.15(1H,bs),
9.33(1H,bs).
(5S)-2-[N-(N-(2,4-dimethoxyphenyl)carbamoyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (K~r) cm~l: 3397, 1711.
CA 02248784 1998-09-11
W O 97/49703 PCT/JP971021S7
H-NMR (CDC13) ~: 1.45(3H,d,J=7.2Hz), 3.37(3H,s),
3.79(3H,s), 3.89(3H,s), 3.92(1H,m), 5.07(1H,d,J=2.8Hz),
6.42-6.47(2H,m), 7.14-7.26(3H,m), 7.39(lH,d,J=7.4Hz),
7.69(1H,d,J=6.8~z), 7.90(1H,d,J=9.2Hz), 8.14(1H,bs),
11.45(lH,bs).
(5s)-2-[N-(N-(7-ethoxycarbonylheptyl)carbamoyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3235, 1716.
lH-NMR (CDC13) ~: 1.17-1.32(9H,m), 1.45(3H,d,J=7.4Hz),
1.50-1.72(4H,m), 2.30(2H,t,J=7.4Hz), 3.20(2H,m),
3.28(3H,s), 4.13(2H,q,J=7.0Hz), 5.02(1H,d,J=3.0Hz), 7.10-
7.25(3H,m), 7.38(1H,d,J=7.6Hz), 7.65(1H,d,J=7.8Hz),
8.34(1H,bs), 9.18(1H,bs).
Example 6
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-(4-
trifluromethylbenzoyl)-N-methyl3amino-2-oxazolin-4-one
To a well stirred mixture of indolmycin (150 mg),
triethylamine (325 ~1), 4-dimethylaminopyridine (39.8 mg),
and tetrahydrofuran (10 ml) under the ice cooling, 4-
trifluoromethylbenzoylchloride (260 ~1) was added. The
mixture was stirred for 30 minutes at 0~C and ethyl acetate
was added. The whole was washed with water, saturated
aqueous sodium hydrogen carbonate solution and brine
respectively and then the ethyl acetate solution was dried
over MgS04. Removal of the organic solvent gave a residue,
which was subjected to silica-gel chromatography. Elution
with hexane-acetone (4:1) provided the titled compound (176
mg, 70.4%). m.p. 146-148~C.
IR (KBr) cm~l: 3390, 1749, 1714.
H-NMR (CDC13) ~: 1.42(3H,d,J=7.2Hz), 3.41~3H,s), 3.77-
3.89(1H,m), 4.92(1H,d,J=2.8Hz), 6.64(1H,d,J=2.0Hz), 7.13-
7.56(8H,m), 8.03(1H,bs).
3~ EXample 7
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/021S7
Following the same procedure as described in the
Example 6, the following compounds were prepared.
- (SS)-5-[~lR)-l-(indol-3-yl)ethyl]-2-(N-(2-
trifluoromethylbenzoyl)-N-methyl]amino-2-oxazolin-4-one,
- 5 IR (KBr) cm~l: 3287, 1748, 1717.
H-NMR (CDC13) ~: 1.23(3H,d,J=7.2Hz), 3.51(3H,s),
3.68(1H,m), 4.87(1H,d,J=2.8Hz), 6.60(1H,d,J=1.8Hz), 7.08-
7.26(3H,m), 7.36-7.61(5H,m), 8.02(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(3-
trifluoromethylbenzoyl)-N-methyl]amino-2-oxazolin-4-one,
IR (KBr) cm~1: 3335, 1715.
H-NMR (CDC13) ~: 1.37(3H,d,J=7.2Hz), 3.41(3H,s),
3.78(1H,m), 4.91(1H,d,J=3.4Hz), 6.62(1H,d,J=2.2Hz), 7.11-
7.40(5H,m), 7.51(1H,d,J=8.0Hz), 7.73(1H,d,J=7.4Hz),
7.78(1H,s), 8.00(1H,bs).
(5S)-2-[N-(4-fluorobenzoyl)-N-methyl]amino-5-[(lR)-l-
(indol-3-yl)ethyl3-2-oxazolin-4-one,
IR (KBr) cm~l: 3300, 1741, 1707.
lH-NMR (CDC13) ~: 1.44(3H,d,J=7.4Hz), 3.38(3H,s), 3.76-
3.gO(lH,m), 4.93(1H,d,J=3.0Hz), 6.76(1H,d,J=2.4Hz), 6.92-
7.00(2H,m), 7.09-7.27(4H,m), 7.40(1H,d,J=8.0Hz),
7.56(1H,d,J=7.2Hz), 8.13(1H,bs).
(5S)-2-[N-(4-chlorobenzoyl)-N-methyl]amino-5-[(lR)-l-
(indol-3-yl)ethyl~-2-oxazolin-4-one,
IR (KBr) cm~l: 3296, 1746, 1705.
lH-NMR (CDC13) ~: 1.42(3H,d,J=7.4Hz), 3.38(3H,s), 3.75-
3.86(1H,m), 4.93(1H,d,J=3.0Hz), 6.70(1H,d,J=2.6Hz), 7.09-
7.28(6H,m), 7.40(1H,d,J=7.8Hz), 7.55(1H,d,J=8.0Hz),
8.09(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(4-
methylbenzoyl)-N-methyl~amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3300, 1744, 1703.
H-NMR (CDC13) ~: 1.36(3H,d,J=7.4Hz), 2.39(3H,s),
3.38(3H,s), 3.71-3.83(1H,m), 4.89(1H,d,J=3.0Hz),
6.59(1H,d,J=2.2Hz), 7.06-7.38(7H,m), 7.50(1H,d,J=8.4Hz),
8.00(lH,bs).
CA 02248784 1998-09-11
W097l49703 PCT/~97102157
(5s)-S-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(4-
methoxybenzoyl)-N-methyl]amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3299, 1744, 1701.
lH-NMR (CDC13) ~: 1.39(3H,d,J=7.4Hz), 3.38(3H,s),
3.82(3H,s), 4.91(1H,d,J=2.8Hz), 6.67(1H,d,J=2.2Hz),
6.81(2H,d,J-8.8Hz), 7.06-7.26(3E,m), 7.36(2H,d,J=8.8Hz),
7.52(1H,d,J=7.4Hz), 8.01(1H,bs).
(5S)-2-(N-cinnamoyl-N-methyl)amino-S-[(lR)-l-(indol-3-
yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3395, 1753, 1682, 1615.
H-NMR (CDC13) ~: 1.46(3H,d,J=7.2Hz), 3.38(3H,s), 3.88-
3.99(lH,m), 5.14(lH,d,J=3.OHz), 7.06-7.82(12H,m),
7.96(lH,bs).
(SS)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
nicotinoyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3277, 1748, 1703.
H-NMR (CDC13) ~: 1.48(3H,d,J=7.4Hz), 3.37(3H,s), 3.79-
3.92(1H,m), 4.95(1H,d,J=3.4Hz), 6.80(1H,d,J=2.2Hz), 7.09-
7.S8(6H,m), 8.12(1H,d,J=2.2Hz), 8.40(1H,bs),
8.63(1H,dd,J=1.8&4.8Hz).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
phenylacetyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3300, 1724.
lH-NMR (CDC13) ~: 1.45(3H,d,J=7.0Hz), 3.25(3H,s), 3.85-
3.99(1H,m), 4.10(1H,d,J=16.6Hz), 4.25(1H,d,J=16.6Hz),
5.05(1H,d,J=2.8Hz), 6.98-7.39(9H,m), 7.63(1H,d,J=8.6Hz),
8.18(1H,bs).
(ss)-s-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-(2
thiophene)carbonyl]amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3289, 1739, 1672.
H-NMR (CDC13) ~: 1.48(3H,d,J=7.4Hz), 3.37(3H,s), 3.81-
3.90(1H,m), 4.99(1H,d,J=3.0Hz), 6.87-7.66(8H,m),
8.08(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-(2-
thienyl~acetyl]amino-2-oxazolin-4-one,
IR (K3r) cm~l: 3298, 1726.
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157
H-NMR (CDC13) ~: 1.49(3H,d,J=7.2Hz), 3.26(3H,s), 3.88-
3.99(1H,m), 4.34(1H,d,J=17.6Hz), 4.51(1H,d,J=17.6Hz),
5.09(1H,d,J=3.2Hz), 6.80-7.22(6H,m), 7.38(1H,d,J=7.4Hz),
7.64(1H,d,J=7.6Hz), 8.17(1H,bs).
(5S)-2-(N-heptanoyl-N-methyl)amino-5-[(lR)-l-(indol-3-
yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3275, 1732.
H-NMR (CDC13) ~: 0.88~3H,t,J=6.2Hz), 1.18-1.33(6H,m),
1.47(3H,d,J=1.2Hz), 1.55-1.63(2H,m), 2.84(2H,q,J=5.4Hz),
3.26(3H,s), 3.87-3.99(1H,m), 5.07(1H,d,J=3.0Hz), 7.09-
7.25(3H,m), 7.38tlH,d,J=7.4Hz), 7.65(1H,d,J=7.4Hz),
8.14(1H,bs).
(5S)-2-(N-cyclohexylcarbonyl-N-methyl)amino-5-[tlR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3320, 1717.
lH-NMR (CDC13) ~: 1.11-1.39(6H,m), 1.50(3H,d,J=7.2Hz),
1.53-1.83(4H,m), 3.23(3H,s), 3.46-3.60(lH,m), 3.85-
3.99(1H,m), 5.06(1H,d,J=3.0Hz), 7.09-7.24(3H,m),
7.37(lH,d,J=7.OHz), 7.65(lH,d,J=7.6Hz), 8.11(lH,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
pivaloyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3287, 1736, 1624.
H-NMR (CDC13) ~: 1.12(9H,s), 1.47(3E,d,J=7.4~z),
3.16(3H,s), 3.90(1H,m), 5.02(1H,d,J=3.0Hz), 7.08-
7.26(3H,m), 7.36(1H,d,J=7.2Hz), 7.64(1H,d,J=7.2Hz),
8.27(lH,bs).
(5s)-2-(N-acetyl-N-methyl)amino-5-[(lR)-l-(indol-3
yl)ethyl~-2-oxazolin-4-one,
IR (KBr) cm~l: 3378, 1750, 1722.
lH-NMR (CDC13) ~: 1.47(3H,d,J=7.0Hz), 2.48(3H,s),
3.25(3H,s), 3.94(1H,m), 5.08(1H,d,J=3.4Hz), 7.09-
7.22(3H,m), 7.38(1H,d,J=7.4Hz), 7.65(1H,d,J=8.0Hz),
8.15(lH,bs).
(5s)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-isobutyl-N
methyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3300, 1736, 1725.
CA 02248784 l998-09-ll
W O 97/49703 PCTtJP97/02157- -
74
H-NMR (CDC13) ~: 0.86(3H,d,J=7.OHz), 1.07(3H,d,J=6.8Hz),
1.50(3H,d,J=7.4Hz), 3.24(3H,s), 3.86(1H,m), 3.94(1H,m),
5.05(1H,d,J=3.0~z), 7.09-7.23(3H,m), 7.36(1H,d,J=7.4Hz),
7.64(1H,d,J=7.4Hz), 8.13(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
propionyl)amino-2-oxazolin-4-one,
IR (KBr) cm 1: 3381, 1736, 1723.
H-NMR (CDC13) ~: 1.05(3H,t,J=7.2Hz), 1.47(3H,d,J=7.2Hz),
2.87(2H,m), 3.26(3H,s), 3.88-4.00(1H,m),
5.06(1H,d,J=3.0Hz), 7.09-7.25(3H,m), 7.38(1H,d,J=7.4Hz),
7.64(1H,d,J=7.4Hz), 8.13(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
palmitoyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3370, 1726.
lH-NMR (CDC13) ~: 0.88(3H,t,J=6.6Hz), 1.16-1.19~26H,m),
1.46(3H,d,J=7.2Hz), 2.82(2H,m), 3.25(3H,s), 3.91(1H,m),
5.06(lH,d,J=3.2Hz), 7.09-7.21(3H,m), 7.37(lH,d,J=7.4Hz),
7.64(1H,d,J=7.8Hz), 8.12(1H,bs).
Example 8
(5S)-2-[N-(2-benzyloxybenzoyl)-N-methyl]amino-5-[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one
To a well stirred mixture of indolmycin (400 mg),
triethylamine (868 ~1) and 4-dimethylaminopyridine (106 mg)
in tetrahydrofuran (20 ml) was added 2-
benzyloxybenzoylchloride (1.15 g) under the ice cooling.
The mixture was stirred for 40 minutes at 0~C and ethyl
acetate was added. The whole was washed with water,
saturated aqueous sodium hydrogen carbonate solution and
brine respectively and then the ethyl acetate solution was
dried over MgSO4. Removal of the organic solvent gave a
residue, which was subjected to silica-gel chromatography.
Elution with hexane-acetone (5:1) provided the titled
compound (534 mg, 73.3%).
IR (KBr) cm~l: 3303, 1744, 1701.
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157 - -
H-NMR (CDC13) ~: 1.15(3H,d,J=7.0Hz), 3.40(3H,s),
3.65(1H,m), 4.68(1H,d,J=2.6Hz), 4.97(2H,s),
6.28(1H,d,J=2.6HZ), 6.89(1H,d,J=8.4Hz), 7.04-7.50(12H,m),
7.85(1H,bs).
Example 9
(SS)-2-[N-(2-hydroxybenzoyl)-N-methyl]amino-5-[(lR)-1-
(indol-3-yl)ethyl]-2-oxazolin-4-one
To a solution of (5S)-2-[N-(2-benzyloxybenzoyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one
(420 mg) in tetrahydrofuran (10 ml) was added 10%
palladium-carbon (300 mg). The whole was subjected to
hydrogenation and then filtered to remove the catalyst.
The filtrate was distilled off to give a residue, which was
treated with ether to provide the titled compound (104 mg,
30.7%) as a crystal.
IR (KBr) cm~l: 3430, 3250, 1752, 1649.
H-NMR (DMSO-d6) ~: 1.38(3~,d,J=7.4Hz), 2.59(3H,s),
3.56(1H,m), 4.77(1H,d,J=3.0Hz), 6.89-7.84(9H,m),
10.62(1H,bs), 10.87(1H,bs).
Example 10
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-(l-
piperidinocarbonyl)-N-methyl]amino-2-oxazolin-4-one
To a mixture of indolmycin (150 mg) and triethylamine
(324 ~1) in tetrahydrofuran (7.0 ml) was added 4-
nitrophenyl chloroformate (353 mg) at 0~C. The mixture was
stirred for 15 minutes at 0~C and then piperidine (173 ~1)
was added. After the whole was further stirred for 8
minutes at 0~C, ethyl acetate was added. The whole was
washed with water, saturated aqueous sodium hydrogen
carbonate solution and brine respectively and then the
ethyl acetate solution was dried over MgSO4. Removal of
the organic solvent gave a residue, which was subjected to
silica-gel chromatography. Elution with hexane-acetone
(3:1) provided the titled compound (154 mg, 71.6%).
CA 022487X4 1998-09-11
W097/49703 PCT/~s7/021s7-
IR (KBr) cm~l: 3279, 1698.
H-NMR (CDC13) ~: 1.20-1.75(9H,m), 3.09-3.35(5H,m), 3.40-
3.70(2H,m), 3.88(1H,m), 4.99(1H,d,J=2.6Hz), 7.10-
7.19(3H,m), 7.34(1H,d,J=7.4Hz), 7.67(1H,d,J=8.4Hz),
8-17(1H,bs).
Example 11
(5S)-2-[N-(N-benzyloxycarbonyl-L-alanyl)-N-methyl]amino-5-
[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one
To a mixture of N-benzyloxycarbonyl-L-alanine (893 mg)
and triethylamine (1.11 ml) in tetrahydrofuran (10 ml) was
added ethyl chloroformate (381 ~1) at -15~C. The mixture
was stirred for 5 minutes at -15~C and then indolmycin (257
mg) was added. After the whole was further stirred for 8
minutes at 0~C, ethyl acetate was added. The whole was
warmed up to room temperature and stirred for 40 minutes.
Ethyl acetate was added to the mixture. The mixture was
washed with water, saturated aqueous sodium hydrogen
carbonate solution and brine respectively and then the
ethyl acetate solution was dried over MgSO4. Removal of
the organic solvent gave a residue, which was subjected to
silica-gel chromatography. Elution with hexane-acetone
(4:1) provided the titled compound (387 mg, 83.3%).
IR (KBr) cm~l: 3233, 1715.
lH-NMR (CDCl3) ~: 1.26-1.60(6H,m), 2.20(3H,s), 4.14(1H,m),
5.05-5.24(4H,m), 6.69(1H,s), 7.11-7.36(9H,m), 7.64(1H,bs),
8.11(lH,s).
Example 12
Following the same procedure as described in the
Example 11, the following compounds were prepared.
(5S)-2-[N-(N-benzyloxycarbonylglycyl)-N-methyl]amino-
5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3277, 1717.
lH-NMR (CDC13) ~: 1.55(3H,d,J=7.8Hz), 2.44(3H,s),
3.91(1H,d,J=15.8Hz), 4.01(1H,d,J=15.8Hz), 4.53(1H,m),
CA 02248784 1998-09-ll
WO 97/49703 PCT/JP97/02157 - -
5.11(2H,s), 5.20(1H,m), 6.61(1H,s), 7.15-7.34(9H,m),
7.62(1H,m), 8.13(1H,bs).
(SS)-2-[N-(N-benzyloxycarbonyl-L-leucyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3300, 1717.
H-NMR (CDC13) ~: 0.75(6H,m), 1.44-1.81(6H,m), 2.19(3H,s),
4.14(1H,m), 5.01-5.27(4H,m), 6.67(1H,s), 7.07-7.22(4H,m),
7.33(5H,m), 7.60(1H,m), 8.11~1H,bs).
(5S)-2-[N-(N-benzyloxycarbonyl-L-phenylalanyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3375, 1719.
H-NMR (CDC13) ~: 1.31(lH,d,J=7.4Hz), 1.45(2H,d,J=7.2Hz),
3.13(3H,s), 3.50(2H,m), 4.42(lH,m), 4.52(lH,m), 5.08-
5.44(3H,m), 6.70-7.60(15H,m), 8.04(lH,s).
(5S)-2-~N-(N-benzyloxycarbonyl-L-prolyl)-N-
methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3324, 1701.
lH-NMR (CDC13) ~: 1.28(1.SH,d,J=7.2Hz),
1.42(1.5H,d,J=5.2Hz), 1.90(3H,m), 2.39(1H,m), 3.11(1.5H,s),
3.30(1.5H,s), 3.51-3.66(2H,m), 3.89(1H,m), 4.89-5.21(3H,m),
5.40-5.52(1H,m), 7.01-7.36(9H,m), 8.01(0.5H,bs),
8.19(0.SH,bs).
(5S)-2-[N-(N-benzyloxycarbonyl-L-y-benzylglutaminyl)-
N-methyl]amino-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-
one,
IR (KBr) cm~l: 3354, 1719.
H-NMR (CDC13) ~: 1.56(3H,m), 1.75(2H,m), 2.17(3H,s),
2.40(2H,m), 3.73(lH,m), 4.25(lH,m), 5.00-5.22(5H,m), 7.11-
7.34(14H,m), 7.64(1H,m), 8.10(1H,bs)
Example 13
(SS)-2-[N-(N-(L-alanyl)-N-methyl]amino-5-[(lR)-l-(indol-3-
yl)ethyl]-2-oxazolin-4-one
To a solution of (5S)-2-[N-(N-benzyloxycarbonyl-L-
alanyl)-N-methyl]amino-5-[(lR)-1-(indol-3-yl)ethyl]-2-
oxazolin-4-one (387 mg) in tetrahydrofuran (4 ml) was added
CA 02248784 1998-09-ll
W O 97/49703 PCT/JPg7/021~7
78
10% palladium-carbon (100 m~). The whole was subjected to
hydrogenation and then filtered to remove the catalyst.
The filtrate was distilled off to give a residue, which was
treated with ether to provide the titled compound (141 mg,
51.6%) as a crystal.
IR (KBr) cm~l: 3372, 3287, 1736, 1633.
H-NMR (CDC13) ~: 1.26(3H,d,J=7.0Hz), 1.52(3H,d,J=7.0Hz),
3.13(3H,s), 3.80(1H,m), 4.22(1H,q,J=7.0Hz), 4.53(1H,bs),
7.07-7.22(2H,m), 7.36(1H,d,J=7.2Hz), 7.72(1H,d,J=7.0Hz),
8.02(1H,bs), 9.O9(lH,bs).
Example 14
Following the same procedure as described in Example
13, the following compounds were prepared.
- 15 (5S)-2-(N-glycyl-N-methyl)amino-5-[(lR)-l-(indol-3-
yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3333, 1748, 1630.
H-NMR (CDC13) ~: 1.27(3H,d,J=7.2Hz), 3.14(3H,s),
3.80(lH,m), 4.15(2H,s), 4.54(lH,d,J=2.8Hz), 7.08-
7.22(2H,m), 7.36(1H,d,J=7.2Hz), 7.72(1H,d,J=7.4Hz),
8.03(1H,bs), 9.02(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-L-leucyl-N-
methyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3368, 1759, 1644.
lH-NMR (CDC13) ~: 1.00(6H,d,J=4.8Hz), 1.27(3H,d,J=7.0Hz),
1.63(1H,m), 1.77(2H,m), 3.13(3H,s), 3.80(1H,m),
4.18(1H,dd,J=3.8&9.2HZ), 4.52(1H,d,J=2.6Hz), 7.07-
7.21(2H,m), 7.36(1H,d,J=7.4Hz), 7.72(1H,d,J=7.4Hz),
8.02(1H,bs), 9.18(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-L-phenylalanyl-
N-methyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3380, 1707, 1637.
H-NMR (CDC13) ~: 1.22(3H,d,J=7.0Hz),
2.93(1H,dd,J-14.0&8.4Hz), 3.04(3H,s),
3.32(lH,d,J=14.0&4.OHz), 3.95(lH,m),
CA 02248784 1998-09-11
W O 97/49703 PCT/JPg7/02157 - -
79
4.37(1H,dd,J=8.4&4.0Hz), 7.07-7.41(9H,m),
7.70(1H,d,J=8.0Hz), 8.01(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-L-prolyl-N-
methyl)amino-2-oxazolin-4-one,
IR (KBr) cm~l: 3289, 1707.
H-NMR (CDC13) ~: 1.51(3H,d,J=7.4Hz), 1.89(2H,m),
2.06(2H,m), 2.78(3H,s), 3.00(lH,m), 3.25(lH,m), 3.80(2H,m),
4.83(1H,d,J=3.2Hz), 6.59(1H,s), 7.09-7.21(3H,m),
7.38(1H,d,J=7.4Hz), 7.64(1H,d,J=7.8Hz), 8.13(1H,bs).
(5S)-2-(N-L-glutaminyl-N-methyl)amino-5-[(lR)-1-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3299, 1724, 1623.
H-NMR (DMSO-d6) ~: 1.18(3H,d,J=6.8Hz), 2.03(2H,m),
2.22(2H,m), 2.99(3H,s), 3.66(1H,m), 4.27(2H,m), 6.91-
7.12(3H,m), 7.32(1H,d,J=7.4Hz), 7.58(1H,d,J=6.6Hz),
9.57(1H,bs).
Example 15
(SS)-5-[(lR)-l-(indol-3-yl)ethyl-2-(N-methanesulfonyl-N-
methyl)amino-2-oxazolin-4-one
To a mixture of indolmycin (100 mg) and triethylamine
(217 ~1) in tetrahydrofuran (5 ~1) was added
methanesulfonylchloride (90.3 ~1) at -30~C. The whole was
stirred for 5 minutes at -30~C and then further stirred at
0~C for 1 hour. Ethyl acetate was added to the mixture.
The mixture was washed with water, saturated aqueous sodium
hydrogen carbonate solution and brine respectively and then
the ethyl acetate solution was dried over MgSO4. Removal
of the organic solvent gave a residue, to which was added
isopropylether to provide the titled compound (74 mg,
57.0%)-
IR (KBr) cm~l: 3303, 1748.
H-NMR (CDC13) ~: 1.53~3H,d,J=7.4Hz), 2.88(3H,s),
3.33(3H,s), 3.88-3.97(1H,m), 5.06(1H,d,J=3.4Hz), 7.10-
7.25(3H,m), 7.37(1H,d,J=7.0Hz), 7.67(1H,d,J=7.8Hz),
8.28(1H,bs).
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157- -
Example 16
Following the same procedure as described in ~xample
15, the following compounds were prepared.
(5S)-2-(N-benzenesulfonyl-N-methyl)amino-5-[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one
IR (KBr) cm~l: 3300, 1748.
lH-NMR (CDC13) ~: 1.23(3H,,d,J=7.2Hz), 3.54(3H,s), 3.76-
3.89(lH,m), 4.99(lH,d,J=2.6Hz), 7.15-7.78(10H,m),
10 8.12(1H,bs).
Example 17
5-[1-(6-fluoroindol-3-yl)ethyl]-2-methylamino-2-oxazolin-4-
one
To a solution of acetoaldehyde (1.0 g) in toluene (7
ml) was added isopropylamine (1.3 g). The mixture was
dried over magnesium sulfate and filtered. The filtrate
was added to a solution of 6-fluoroindole (3.32 g) in
acetic acid (20 ml) under ice cooling. The whole was
stored at freezer for 3 days and was poured into the ice
water. The mixture was neutralized with 25% ammonia water.
The whole was extracted with ethyl acetate. The extract
was washed with brine and then dried over MgSO4. Removal
of the solvent gave a residue, to which a mixture of ethyl
acetate and ethyl ether was added to provide 6-fluoro-3-(1-
isopropylamino)ethylindole (1.31 g) as a crystal.
H-NMR (CDC13) ~: 1.07(6H,m), 1.49(3H,d,J=6.6Hz),
2.85(1H,m), 4.23(1H,q,6.6Hz), 6.80-7.10(3H,m),
7.62(1H,dd,J=5.4&8.4Hz), 8.12(1H,bs).
To a mixture of 6-fluoro-3-(1-
isopropylamino)ethylindole (435 mg), benzyl 2-
dimethylamino-4-oxo-2-oxazolin-5-carboxylate (518 mg) in
acetonitrile (15 ml) was added tri-n-butylphosphine (492
~1). The mixture was refluxed for 2.5 hours. Removal of
the solvent gave a residue, to which a mixture of ethyl
acetate and ethyl ether in a ration of 10:1 was added to
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/021S7- -
81
provide benzyl 2-dimethylamino-5-[1-(6-fluoroindol-3-
yl)ethyl]-4-oxo-2-oxazolin-S-carboxylate (509 mg) as a
crystal.
lH-NMR (CDC13) ~: 1.28(2H,d,J=7.2Hz), 1.51(lH,d,J=7.2Hz),
2.84-3.26(6H,m), 4.24(lH,m), 4.99(1.4H,ABq,J=4.8Hz),
5.32(0.7H,ABq,J=4.8Hz), 6.70-7.70(9H,m), 8.14(0.7H,bs),
8.45(0.35H,bs).
Benzyl 2-dimethylamino-5-[1-(6-fluoroindol-3-
yl)ethyl3-4-oxo-2-oxazolin-5-carboxylate (500 mg) was
dissolved into 15 ml of a solution of ethanol and
tetrahydrofuran in a ration of 5:1. 10% palladium-carbon
(170 mg) was added. The whole was subjected to
hydrogenation under normal temperature and normal pressure
for 1.5 hours. The whole was stirred at 80~C under
nitrogen atmosphere for 1 hour and then filtered to remove
the catalyst. The filtrate was concentrated to give 2-
dimethylamino-5-tl-(6-fluoroindol-3-yl)ethyl]-2-oxazolin-4-
one (340 mg).
lH-NMR (CDC13) ~: 1.43(1.2H,d,J=7.2Hz),
1.62(1.8H,d,J=7.2Hz), 2.97-3.06(6H,m), 3.60-3.80(lH,m),
4.90(0.6H,d,J=1.5Hz), 4.97(0.4H,d,J=1.5Hz), 6.85(1H,m),
6.90-7.30(2H,m), 7.58(1H,m), 8.66(0.4H,bs), 8.67(0.6H,bs).
2-Dimethylamino-5-[1-(6-fluoroindol-3-yl)ethyl]-2-
oxazolin-4-one (340 mg) was dissolved into methylamine (5
ml) at -10~C. The mixture was stirred for 3 hours at the
same temperature. The mixture was concentrated to give a
residue, which was subjected to column chromatography.
Elution with hexane-acetone (1:1) provided the titled
compound (254 mg).
IR (KBr) cm~l: 3195, 1733, 1644, 1627.
H-NMR (DMSO-d6) ~: 1.18(0.9H,d,J=7.2Hz),
1.27(0.4H,d,J=7.4Hz), 1.43(1.1H,d,J=7.2Hz),
1.49(0.6H,d,J=7.2Hz), 2.60-2.80(3H,m), 3.40-3.60(1H,m),
4.80-5.00(1H,m), 6.81(1H,m), 7.00-7.20(2H,m), 7.55(1H,m),
8.50-8.70(1H,bs), 10.9-ll.O(lH,m).
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157 - -
82
Example 18
Following the same procedure as described in ~xample
11, the following compounds were prepared.
(5S)-2-[N-(3-benzyloxycarbonylaminopropionyl)-N-
methylamino]-5- E ( lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm 1 3327, 2971, 1715, 1557, 1539, 1456, 1397,
1316, 1246, 1200, 1138.
lH-NMR (CDC13) ~: 1.48(3H,d,J=7.4Hz), 2.76-3.07(2H,m),
3.20(3H,s), 3.30-3.42(2H,m), 3.93(1H,m),
5.06(1H,d,J=3.2Hz), 5.11(2H,s), 7.07-7.22(3H,m), 7.30-
7.40(6H,m), 7.62(1H,d,J=7.8Hz), 8.24(1H,bs).
(SS)-S-[(lR)-l-(indol-3-yl)ethyl]-2-[N-[3-(N-
benzyloxycarbonyl-N-methyl)amino]propionyl-N-methylamino]-
2-oxazolin-4-one,
IR (KBr) cm~l: 3296, 1700-1750, 1646.
H-NMR (CDC13) ~: 1.38-1.50(3H,m), 2.86(1.2H,s),
2.90(1.8H,s), 2.97-3.11(2H,m), 3.16(1.8H,s), 3.22(1.2H,s),
3.4-3.6(2H,m), 3.92(1H,m), 5.04(1H,d,J=2.8Hz), 5.13(2H,s),
6.9-7.4(9H,m), 7.61(1H,d,J=7.4Hz), 8.1-8.3(1H,bs).
(SS)-2-[N-(4-benzyloxycarbonylaminobutyryl)-N-
methylamino]-5-[(lR)-l-(indol-3-yl)ethyl~-2-oxazolin-4-one,
IR (KBr) cm~l: 3337, 2969, 2940, 1711, 1561, 1537, 1454,
1433, 1397, 1314, 1252, 1192.
lH-NMR (CDC13) ~: 1.48(3H,d,J=7.4Hz), 1.64(2H,m),
2.76(2H,m), 3.12(2H,m), 3.23(3H,s), 3.93(lH,m),
4.82(1H,bs), 5.05(1H,d,J=3.2Hz), 5.12(2H,s), 7.00-
7.41(9H,m), 7.62(1H,d,J=6.8Hz), 8.35(1H,bs).
(5S)-2-[N-(4-benzylsuccinyl)-N-methylamino]-5-[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
I~ (KBr) cm~l: 3400, 1733, 1558, 1538, 1456, 1432, 1394,
1209, 1166.
H-NMR (CDC13) ~: 1.46(3H,d,J=7.3Hz), 2.52-2.66(2H,m),
2.99-3.34(2H,m), 3.24(3H,s), 3.93(1H,dq,J=7.3&3.0Hz),
5.06(1H,d,J=3.0Hz), 5.12(2H,s), 7.07-7.38(9H,m),
7.63(1H,d,J=7.2Hz), 8.10(1H,bs).
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157
83
(SS)-2-[N-[(2S)-2,5-bis(N-benzyloxycarbonylamino)-
pentanoyl]-N-methylamino-5-[(lR)-l-(indol-3-yl)ethyl-2-
oxazolin-4-one,
IR (KBr) cm~l: 3327, 3064, 3033, 2968, 2939, 2877, 1718,
1523, 1455, 1388, 1345, 1263, 1220.
H-NMR ~CDC13) ~: 1.25-1.55(4H,m), 1.57(3H,d,J=6Hz),
2.33(3H,s)~ 2.70-2.90(2H,m), 4.15-4.30(2H,m), 4.83-
5.29(7H~m)~ 7.07-7.68(15H,m), 8.12(1H,bs).
(5S)-2-[N-[(S)-4-benzyl(N-benzylcarbonylamino)-
aspartyl]-N-methylamino]-5-[(lR)-l-(indol-3-yl)ethyl]-2-
oxazolin-4-one,
IR (KBr) cm~l: 3396, 3064, 3035, 2970, 2937, 1722, 1455,
1423, 1386, 1344, 1311.
lH-NMR (CDC13) ~: 1.36,1.60(total 3H,d,J=7.8Hz),
2.19(3H,s), 2.28-3.18(2H,m), 3.78(1H,m), 4.21(1H,m), 4.29-
4.47(lH,m), 4.99-5.32(5H,m), 6.70-7.47(14H,m),
7.68(1H,d,J=8.2Hz), 8.10(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-[(S)-4-
methyl[N-benzyloxycarbonylamino)-aspartyl~-N-methylamino]-
2-oxazolin-4-one,
IR (KBr) cm~l: 3402, 3060, 3037, 2954, 2937, 1722, 1457,
1436, 1376, 1344, 1313.
H-NMR (CDC13) ~: 1.36,1.61(total 3H,d,J=7.6Hz),
2.20(3H,s)~ 2.82-3.82(3H,m), 3.57,3.60(total 3H,s), 4.18-
4.50(2H,m), 5.02-5.32(3H,m), 6.85-7.36(9H,m),
7.45,7.68(total lH,d,J=7.2Hz), 8.14(1H,bs).
(5S)-2-[N-(2-tert-butyldimethylsilyloxyoctanoyl)-N-
methylamino]-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
lH-NMR (CDC13) ~: -0.1-0.1(6H,m), 0.85(4.5H,s),
0.87(4.5H,s), 1.0-1.2(10H,m), 1.46(3H,t,J=7.0Hz),
3.25(1.5H,s), 3.26(1.5H,s), 3.93(1H,dq,J=3.0&7.0Hz),
5.05(0.5H,d,J=3.OHz), 5.07(0.5H,d,J=3.0Hz), 5.40-
5.48(1H,m), 7.1-7.7(5H,m), 8.13(1H,bs).
(5S)-2-[N-(2-benzyloxyoctanoyl)-N-methylamino]-5-
[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 1810, 1733, 1704, 1634.
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157- -
84
lH-NMR (CDC13) ~: 0.8-0.9(3H,m), 1.1-1.4(9H,m),
1.45(3H,d,J=7.2Hz), 1.5-1.83(1H,m), 3.2-3.3(3H,m),
3.89~1H,dq,J=3.0&7.2Hz), 3.97-4.73(3H,m), 5.03(0.5H,m),
5.11-5.17(0.5H,m), 6.9-7.7(10H,m), 8.11(0.5H,bs),
8.16(0.5H,bs).
(5S)-2-(N-benzyloxyacetyl-N-methylamino)-5-~(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3347, 1733, 1627, 1557.
lH-NMR (CDC13) ~: 1.46(3H,d,J=7.2Hz), 3.26(3H,s),
3.92(1H,dq,J=3.4&7.2Hz), 4.42,4.66(2H,ABq,J=18.0Hz),
4.55(2H,s), 5.05(1H,d,J=3.4Hz), 7.0-7.7(10H,m),
8.09(lH,bs).
(SS)-2-[N-(2-benzyloxy-4-methylpentanoyl)-N-
methylamino]-5-[(lR)-l-(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 1742, 1729, 1557, 1538.
H-NMR (CDC13) ~: 0.66,0.78(each 3H,d,J=6.6Hz),
0.88,0.93(3H,d,J=7.0Hz), 1.3-1.6(3H,m), 3.2-3.3(3H,m),
3.91(1H,m), 3.8-4.6(2.5H,m), 5.00-5.05(1H,m), 5.25(0.5H,m),
7.0-7.7(10H,m), 8.09-8.2(lH,m).
Example 19
Following the same procedure as described in Example
6, the following compounds were prepared.
(5S)-5-~(lR)-l-(indol-3-yl)ethyl]-2-[N-lauroyl-N-
methylamino]-2-oxazolin-4-one,
IR (KBr) cm~l: 3298, 2925, 2854, 1729, 1558, 1538, 1455,
1394, 1195.
H-NMR (CDC13) ~: 0.83(3H,t,J=6.6Hz), 1.23-1.35(16H,m),
1.46(3H,d,J=7.2Hz), 1.51-1.64(2H,m), 2.85(2H,m),
3.25(3H,s), 3.93(1H,m), 5.06(1H,d,J=3.0Hz), 7.09-
7.25(3H,m), 7.37(1H,d,J=7.8Hz), 7.64(1H,d,J=7.8Hz),
8.14(lH,bs).
(5S)-2-[N-chloroacetyl-N-methylamino]-5-[(lR)-l-
(indol-3-yl)ethyl3-2-oxazolin-4-one,
IR (KBr) cm~l: 3383, 2972, 1733, 1558, 1455, 1436, 1398,
1340, 1317, 1203.
CA 02248784 1998-09-11
- W097/49703 PCT/~97/02157
H-NMR (CDC13) ~: 1.52(3H,d,J=7.3Hz), 3.28(3H,s),
3.95(1H,m), 4.48(1H,d,J=16.5Hz), 4.72(1H,d,J=16.5Hz),
5.08(1H,d,J=3.3Hz), 7.08-7.26(3H,m), 7.38(1H,d,J=7.4Hz),
7.62(1H,d,J=7.6Hz), 8.25(1H,bs).
(SS)-2-[N-[2-(5-amino-1,2,4-thiodiazol-3-yl)-2(Z)-
ethoxyiminoacetyl]-N-methylamino]-5-(lR)-l-(indol-3-
yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3400, 2976, 1747, 1714, 1616, 1538, 1455,
1403, 1245, 1224.
lH-NMR (CDC13) ~: 1.14(3H,t,J=7.1Hz), 1.23(3H,m),
3.56(3H,s), 3.79(lH,m), 5.08(lH,m), 6.85-7.25(5H,m),
7.33(1H,d,J=8.2Hz), 7.56(1H,d,J=7.0Hz), 8.60(1H,bs).
(5S)-2-(N-allyloxalyl-N-methylamino)-5-[(lR)-l-(indol-
3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 1746, 1704, 1634, 1615.
H-NMR (CDC13) ~: 1.25-1.45(3H,m), 3.00-3.11(3H,m),
3.76(1H,dq,J=3.4&7.2Hz), 4.90-5.01(3H,m),
5.40(1H,d,J=11.2Hz), 5.51(1H,dd,J=1.2&17.0Hz), 5.95-
6.15(1H,m), 7.1-7.7(5H,m), 8.46(1H,bs).
Example 20
Following the same procedure as described in Example
13, the following compounds were prepared.
(5S)-2-~N-(3-aminopropionyl)-N-methylamino}-5-[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3266, 2969, 2928, 1705, 1622, 1584, 1495,
1456, 1399, 1341, 1308, 1236.
H-NMR (CDC13) ~: 1.27(3H,d,J=7.0Hz), 2.78(2H,t,J=7.0Hz),
3.30(3H,s), 3.56(2H,t,J=7.0Hz), 3.65(2H,bs), 3.77(1H,m),
4.48(1H,m), 7.06-7.21(3H,m), 7.35(1H,d,J=7.6Hz),
7.71(1H,d,J=7.0Hz), 8.08(1H,bs).
(5S)-5-[(lR)-l-(indol-3-yl)ethyl]-2-[N-[(S)-4-
methylaspartyl]-N-methylamino]-2-oxazolin-4-one,
IR (KBr) cm~l: 3352, 2966, 1735, 1643, 1577, 1488, 1457,
1438, 1403, 1322, 1255, 1099.
CA 02248784 l998-09-ll
W 097/49703 PCT/JP97tO2157
86
lH-NM~ (CDC13) ~: 1.27(3H,d,J=7.1Hz),
2.73(1H,dd,J=17.6&9.0Hz), 3.07(1H,dd,J=17.6&3.4Hz),
3.15(3H,s), 3.57(1H,bs), 3.76(3~,s), 3.82(1H,m),
4.46(1H,dd,J=9.0&3.4Hz), 4.54(1H,m), 7.03-7.27(3H,m),
7-36(1H,d,J=7-0Hz), 7-73(1H,d,J=7.4Hz), 8-07(1H,bs),
9.12(1H,bs).
(5S)-2-[N-[(S)-aspartyl]-N-methylamino]-5-[(lR)-l-
(indol-3-yl)ethyl]-2-oxazolin-4-one,
IR (KBr) cm~l: 3400, 1720, 1629, 1560, 1425, 1399, 1342.
lH-NMR (DMSO-d6) ~: 1.17(3H,d,J=7.0Hz), 2.30-2.70(2H,m),
2.99(3H,s), 3.64(lH,m), 4.24-4.38(2H,m), 6.93-7.36(4H,m),
7.60(1H,d,J=8.0Hz), 10.78(1H,bs).
(5S)-5-~(lR)-l-(indol-3-yl)ethyl]-2-[N-methyl-N-(3-
methylaminopropionyl)amino]-2-oxazolin-4-one,
IR (KBr) cm~l: 3244, 1733, 1607.
H-NMR (CDC13) ~: 1.49(2.0H,d,J=7.4Hz),
1.54(1.0H,d,J=7.4Hz), 1.9-2.5(2H,m), 2.59(1.0H,d,J=2.4Hz),
2.71(2.0H,d,J=2.4Hz), 2.97(2.0H,s), 3.00(1.0H,s), 3.4-
3.6(1H,m), 3.6-3.8(1H,m), 3.86(1H,dq,J=3.0&7.4Hz),
4.80(0.33H,bs), 4.98(1H,d,J=3.0Hz), 6.08(0.67H,bs), 7.0-
7.7(5H,m), 8.64(0.67H,bs), 8.70(0.33H,bs).
Example 21
(5S)-2-lN-(2-benzylphthaloyl)-N-methylamino]-5-[(lR)-l-
(indoly-3-yl)ethyl}-2-oxazolin-4-one
To a solution of mono-benzyl phthalic ester (1.60 g)
in toluene (20 ml) was added thionyl chloride (1 ml).
After being stirred for one hour at 80~C, the mixiture was
concentrated under the reduced pressure to afford a
residue. Tetrahydrofuran (20 ml) was added to the residue
and then indolmycin (400 mg) was added at -15~C. After
triethylamine (1.74 ml) was added, the mixture was stirred
for 21 hours at room temperature. The whole was diluted
with ethyl acetate (100 ml). The mixture was washed with
water (35 ml), saturated aqueous sodium hydrogen carbonate
solution (35 ml X3) and brine (35 ml) respectively and the
CA 02248784 1998-09-11
W097/49703 PCT/~97102157
87
dried over MgSO4. Concentration of the solution under the
reduced pressure gave a residue, which was subjected to
column-chromatography. Eluent with ethyl acetate-hexane
was collected and concentrated. The content was solidified
- 5 from hexane. The solid product was collected by filtration
and dried to provide the titled compound (161 mg).
IR (KBr) cm~l: 3400, 1714, 1538, 1455, 1399, 1278, 1222.
H-NMR (CDC13) ~: 1.07(3H,d,J=7.1Hz), 3.44(3H,s),
3.60(1H,m), 4.72(1H,d,J=2.6Hz), 5.17(1H,d,J=12.0Hz),
10 5.26(1H,d,J=12.0Hz), 6.31(1H,d,J=2.4Hz), 7.06-7.52(12H,m),
7.86(1H,bs), 7.95(1H,dd,J=7.6&1.4Hz).
Example 22
(SS)-5-[(lR)-l-(indol-3-yl)ethyl]-2-(N-methyl-N-
trifluoroacetylamino)-2-oxazolin-4-one
Indolmycin (401 mg) was dissolved into tetrahydrofuran
(38 ml). To this solution were added triethylamine (1.5
ml) and trifluoroacetic anhydride (1.0 ml). The whole was
stirred for 1 hour at room temperature. The mixture was
diluted with ethyl acetate (100 ml). The whole was washed
with water, brine and dried over magnesium sulfate. The
solution was concentrated to give a residue, which was
subjected to silica gel chromatography. Elution with
hexane-acetone (1:1) gave the titled compound (375 mg).
25 IR (KBr) cm~1: 1733, 1652, 1634, 1615.
lH-NMR (CDC13) ~: 1.33(0.45H,d,J=7.0Hz),
1.34(2.55H,d,J=7.0Hz), 2.97-3.13(3H,m),
3.77(1H,dq,J=2.2&7.0Hz), 4.85(0.15H,d,J=2.2Hz),
4.97(0.85H,d,J=2.2Hz), 7.3-7.7(5H,m), 8.4-8.51(1H,bs).
Exampale 23
2-methylamino-5-[(5-chloroindol-3-yl)methyl]-2-oxazolin-4-
one
To a solution of 5-chloro-3-formylindole (5.00 g) in
35 tetrahydrofuran (135 ml) was added sodium hydride (60% oil
suspension, 3.53 g). The mixture was stirred for 20
CA 02248784 1998-09-11
- W097/49703 PCT/~97/02157- -
minutes at room temperature. Carbobenzoxychloride (6.64
ml) was added to the mixture. After being stirred for 1
hour at room temperature, the whole was poured into ice
water. The mixture was extracted with ethyl acetate. The
extract was washed with brine and dried over magnesium
sulfate. The solution was concentrated to give a residue,
which was subjected to silica gel chromatography. Elution
with hexane-ethyl acetate (5:1) provided 1-
benzyloxycarbonyl-5-chloro-3-formylindol (5.74 g) as a
10 Crystal.
H-NMR (CDCl3) ~: 5.51(2H,s), 7.36-7.52(6H,m),
8.10(1H,d,J=9.2Hz), 8.27(1H,s), 8.30(1H,d,J=2.2Hz),
10.06(1H,s).
To a solution of l-benzyloxycarbonyl-5-chloro-3-
formylindol (2.00 g) in methanol (13 ml) was added sodiumborohydride (241 mg) at 0~C. After the mixture was stirred
for 15 minutes at the same temperature, ice water was added
to the mixture. Potassium carbonate was added to make the
mixture saturate. The whole was extracted with ether and
the ext~ract was dried over magnesium sulfate.
Concentration of the mixture gave a residue, to which
hexane was added. l-benzyloxycarbonyl-5-chloro-3-
hydroxymethylindol (1.80 g) was obtained as a crystal.
lH-NMR (CDC13) ~: 1.63(1H,t,J=5.0Hz), 4.79(2H,d,J=5.0Hz),
5.44(2H,s), 7.26-7.45(6H,m), 7.63(2H,s),
8.09(1H,d,J=8.8Hz).
To a solution of l-benzyloxycarbonyl-5-chloro-3-
hydroxymethylindole (1.45 g) in dichloromethane (23 ml) was
added thionylchloride (0.797 ~1) at -78~C. After the
mixture was stirred for 1.5 hours at room temperature, the
reaction mixture was concentrated under the reduced
pressure to give a residue. Hexane was added to the
residue. l-benzyloxycarbonyl-5-chloro-3-chloromethylindole
(1.43 g) was obtained as a crystal.
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157 - -
89
lH-NMR (CDC13) ~: 4.70(2H,s), 5.44(2H,s),
7.32(1H,dd,J=8.8&1.8Hz), 7.39-7.46(5H,m),
7.64(1H,d,J=1.8Hz), 7.69(1H,s), 8.10(1H,d,J=8.8Hz).
To a solution of diisopropylamine (1.18 ml) in
tetrahydrofuran (24 ml) was added a 1.6 M solution (5.24
ml) of n-butyllithium in hexane under ice cooling. The
mixture was cooled to -78~C, to which was added 2-
dimethylamino-2-oxazolin-4-one (1.07 g). The mixture was
stirred for 30 minutes at room temperature and cooled again
to -78~C. 1-benzyloxycarbonyl-5-chloro-3-chloromethylindol
(1.40 g) was added. The whole was stirred at -78~C to -40~C
for 30 minutes, at 0~C for 2 hours and at room temperature
for 40 minutes respectively. To the reaction mixture was
added water and the whole was extracted with ethyl acetate.
The extract was dried over magnesium sulfate and
concentrated to give a residue, which was subjected to
silica gel chromatography. Elution with hexane-ethyl
acetate (1:1) gave 5-[(5-chloroindol-3-yl)methyl-2-
dimethylamino-2-oxazolin-4-one (232 mg) as a crystal.
lH-NMR (CDC13) ~: 2.96(3H,s), 2.98(3H,s),
3.22(1H,dd,J=15.8~4.0Hz), 3.45(1H,dd,J=15.8&4.0Hz),
4.98(1H,t,J=4.0Hz), 7.07(1H,s), 7.09(1H,d,J=8.8Hz),
7.29(1H,d,J=8.8Hz), 7.59(1H,s), 8.64(1H,bs).
To [(5-chloroindol-3-yl)methyl~-2-dimethylamino-2-
oxazolin-4-one (200 mg) was added methylamine (20 ml). The
mixture was refluxed at -6~C for 1 hour. Removal of
methylamine gave a residue, to which was added ether. The
titled compound (120 mg) was obtained as a crystal.
IR (KBr) cm~l: 2986, 1641, 1413, 1390, 1304, 1242, 1103.
lH-NMR (DMSO-d6) ~: 2.73(3H,s), 3.01(1H,m), 3.19(1H,m),
4.95(1H,m), 7.03(1H,d,J=8.8Hz), 7.20(1H,s),
7.34(1H,dd,J=8.8&1.8Hz), 7.58(1H,d,J=1.8Hz), 8.31(1H,bs).
Example 24
Following the same procedure as described in Example
23, the following compounds were prepared.
CA 02248784 1998-09-11
- W O 97/49703 PCT/JP97/021S7
2-methylamino-5-[~2-methylindol-3-yl)methyl]-2-
oxazolin-4-one,
IR (KBr) cm~l: 2912, 1655, 1508, 1408, 1305, 1251, 1238,
746.
lH-NMR (DMSO-d6) ~: 2.50(3H,s), 2.70(3H,s), 2.93-
3.20(2H,m), 4.86(lH,m), 6.86-7.00(2H,m),
7.20(1H,d,J=7.2Hz), 7.40(1H,d,J=6.9Hz),-8.48(1H,bs),
10.76(1H,bs).
5-[(5-benzyloxyindol-3-yl)methyl]-2-methylamino-2-
oxazolin-4-one,
IR (KBr) cm~l: 1667, 1640, 1485, 1412, 1304.
lH-NMR (DMSO-d6) ~: 2.68(1.5H,s), 2.99(1H,m), 3.21(1H,m),
4.95(1H,dd,J=7.0&3.4Hz), 6.79(1H,d,J=8.8Hz), 7.08-
7.52(8~,m), 10.78(1H,bs).
- 15 2-methylamino-5-[(5,6-methylenedioxyindoly-3-
yl)methyl]-2-oxazolin-4-one,
H-NMR (DMSO-d6) ~: 2.73(3H,d,J=5.0Hz), 2.96-3.15(2H,m),
4.89-4.91(1H,m), 5.92(2H,s), 6.85(1H,s), 6.96(1H,s),
6.99(1H,s), 8.58(1H,bs).
Example 25
2-methylamino-5-(lH-pyrro[2,3-b~pyridin-3-yl)methyl-2-
oxazolin-4-one
To a solution of diisopropylamine (0.75 ml) in
tetrahydrofuran (35 ml) was added 1.6M solution (3.35 ml)
of n-butyllithium in hexane under ice cooling. The whole
was cooled to -78~C. 2-dimethylamino-2-oxazolin-4-one (686
mg) was added and then the whole was stirred for 30 minutes
at room temperature. The mixture was cooled again to -
78~C, to which was added 3-chloromethyl-lH-pyrro[2,3-
b]pyridine (230 mg). The whole was stirred for 4 hours at
room temperature. To the reaction mixture was added water
and the whole was extracted with ethyl acetate. The
extract was dried over magnesium sulfate and concentrated
to give a residue, which was subjected to silica gel
chromatography. Elution with ethyl acetate-ethanol (10:1)
CA 02248784 1998-09-ll
W O 97/49703 PCTtJP97/02157 - -
91
provided 2-dimethylamino-5-(lH-pyrro[2,3-b]pyridin-3-
yl)methyl-2-oxazolin-4-one (51 mg).
H-NMR (CDC13) ~: 2.95(3H,s), 3.03(3H,s),
3.26(1H,dd,J-15.9&5.5Hz), 3.48(1H,dd,J=15.9&4.3Hz),
4.97(1H,dd,J-5.5&4.3HZ), 7.09(1H,dd,J=7.9&4.8Hz),
7.24(1H,s), 7.98(1H,dd,J=7.9&1.5Hz), 8.29(1H,J=4.8&1.5Hz),
9.64(1H,bs).
2-dimethylamino-5-(lH-pyrro[2,3-b]pyridin-3-yl)methyl-
2-oxazolin-4-one (50 mg) was added to methylamine (1 ml).
After the reaction mixture was refluxed for 1 hour,
methylamine was removed to give a residue, which was
subjected to silica gel chromatography. Eluent with ethyl
acetate-ethanol was concentrated to give a residue, which
was solidified using chloroform. The solid was washed with
diethyl ether and dried under the reduced pressure to give
the titled compound (32 mg).
IR (KBr) cm~l: 3215, 1645, 1516.
H-NMR (DMSO-d6) ~: 2.71,2.73(total 3H,each s), 2.99-
3.32(2H,m), 4.97(lH,dd,J=6.7&3.9~z),
7.03(1H,dd,J=7.8&4.7Hz), 7.24(1H,d,J=2.1Hz),
7.96(1H,d,J=7.8Hz), 8.18(1H,d,J=4.7Hz), 8.58(1H,bs),
11.43(lH,bs).
Example 26
Following the same procedure as described in Example
25, the following compounds were prepared.
5-(bezo[b]thiophen-3-yl)methyl-2-methylamino-2-
oxazolin-4-one,
IR (KBr) cm~l: 3210, 1768, 1678.
lH-NMR (DMSO-d6) ~: 2.68,2.79(total 3H,each s),
3.18(1H,dd,J=15.7&8.4Hz), 3.42(1H,m), 5.06(1H,m), 7.35-
7.45(2H,m), 7.51(1H,s), 7.82-7.99(2H,m), 8.69(1H,bs).
5-[(1-benzylindol-3-yl)methyl]-2-methylamino-2-
oxazolin-4-one,
IR (KBr) cm~l: 1663, 1508, 1410, 1402, 1298, 729.
CA 02248784 1998-09-11
- W O 97/49703 PCT/JP97/02157
H-NMR (DMSO-d6) ~: 2.68(1H,s), 2.72(2H,s), 3.07(1H,m),
3.30(1H,m), 4.98(1H,dd,J=6.2&2.4Hz), 5.37(2H,s), 6.96-
7.38(9H,m), 7.56(1H,d,J=7.0Hz), 8.55(1H,bs).
Example 27
Following the same procedure as described in Example
17, the following compounds were prepared.
S-[(1-(4-methylindol-3-yl)ethyl]-2-methylamino-2-
oxazolin-4-one,
IR (KBr) cm~l: 3183, 1733, 1623.
H-NMR (CDC13) ~: 1.2S-1.50(3H,m), 2.69-3.0(3H,m), 4.05-
4.25(1H,m), 4.86(0.5H,d,J=3.6Hz), 4.96(0.5H,d,J=2.6Hz),
6.84(1H,m), 6.98-7.25(3H,m), 8.29(0.5H,bs), 8.38(0.5H,bs),
9.30(lH,bs).
5-[(1-(4-benzyloxyindol-3-yl)ethyl]-2-methylamino-2-
oxazolin-4-one,
IR (KBr) cm~1: 3199, 1733, 1652, 1615.
H-NMR (CDC13) ~: 1.22-1.37(3H,m), 2.82-3.0(3H,m), 4.1-
4.4(1H,m), 5.08(0.5H,d,J=4.8Hz), 5.19(1.5H,m), 5.26(1H,s),
6.56(1H,m), 6.9-7.1(3H,m), 7.29-7.6(5H,m), 8.23(0.5H,bs),
8.37(0.5H,bs), 9.25(1H,bs).
2-methylamino-5-[1-(7-methylindol-3-yl)ethyl]-2-
oxazolin-4-one,
IR (KBr) cm~1: 3357, 3214, 1733, 1652, 1615.
lH-NMR (DMSO-d6) ~: 1.18(0.96H,d,J=7.4Hz),
1.25(0.45H,d,J=7.OHz), 1.42(0.96H,d,J=7.2Hz),
1.50(0.63H,d,J=7.6Hz), 2.41(1.5H,s), 2.44(1.5H,s), 2.65-
2.8(3H,m), 3.5-3.7(lH,m), 4.8-4.95(lH,m), 6.8-6.9(2H,m),
7.0-7.15(1H,m), 7.35-7.45(1H,m), 8.55(1H,bs).
5-[1-(4-methoxycarbonylindoly-3-yl)ethyl]-2-
methylamino-2-oxazolin-4-one,
IR (KBr) cm~l: 3278, 3203, 1713, 1635.
H-NMR (DMSO-d6) ~: 1.06(1.33H,d,J=7.OHz),
1.14(0.67H,d,J-7.0Hz), 1.32(0.67H,d,J=7.0Hz),
1.41(0.33H,d,J=7.6Hz), 2.7-2.85(3H,m), 3.86(1.08H,s),
CA 02248784 1998-09-11
W O 97/49703 PCTIJP97/02157
3.91(1.92H,s), 3.95-4.2(1H,m), 4.65-4.85(1H,m), 7.0-
7.2(1H,m), 7.3-7.5(2H,m), 7.55-7.65(1H,m), 11.40(1H,bs).
5-[1-(4-isopropylindol-3-yl)ethyl]-2-methylamino-2-
oxazolin-4-one.
IR (KBr) cm~l: 3266, 3216, 1725, 1634.
H-NMR (DMSO-d6) ~: 1.1-1.5(9H,m), 2.75-2.9(3H,m), 3.6-
3.9(1H,m), 4.8-4.95(1H,m), 6.8-7.1(2H,m3, 7.1-7.25(2H,m),
8.67(1H,bs).
Example 28
5-[1-(4-hydroxyindol-3-yl)ethyl]-2-methylamino-2-oxazolin-
4-one
5-[1-(4-benzyloxyindol-3-yl)ethyl]-2-methylamino-2-
oxazolin-4-one (149 mg) was dissolved in a mixture of
ethanol and tetrahydrofuran (5:1.6 ml). 10% palladium-
carbon (45 mg) was added and the whole was hydrogenated for
24 hours at room temperature under normal pressure. The
catalyst was removed from the mixture by filtration and 10%
palladium-carbon (45 mg) was added again. The whole was
hydrogenated for additional 24 hours at room temperature
under normal pressure. The reaction mixture was filtered
to remove the catalyst. The filtrate was concentrated to
give a residue, which was subjected to silica gel
chromatography. Elution with hexane-acetone (1:1) provided
the titled compound (72 mg).
IR (KBr) cm~l: 3189, 1733, 1698, 1615.
H-NMR (DMSO-d6) ~: 1.0-1.15(1.92H,m), 1.30-1.35(1.08H,m),
2.7-2.85(3H,m), 3.7-4.0(1H,m), 4.92(0.12H,d,J=6.0~z),
5.02(0.31H,d,J=6.0Hz), 5.08(0.20H,d,J=2.2Hz),
5.13(0.37H,d,J=2.2Hz), 6.32(1H,d,J=5.2Hz), 6.7-7.0(3H,m),
8.58(1H,bs), 9.40(1H,bs).
Example 29
5-[1-(4-methoxyindol-3-yl)ethyl]-2-methylamino-2-oxazolin-
4-one
CA 02248784 1998-09-11
- W O 97149703 PCTIJP97/02157 - -
94
5-[1-(4-benzyloxyindol-3-yl)ethyl]-2-dimethylamino-2-
oxazolin-4-one (268 mq) was dissolved in a mixture of
ethanol and tetrahydrofuran (7:3.10 ml). 10~ palladium-
carbon (80 mg) was added and the whole was hydrogenated for
S hours at room temperature under normal pressure. The
catalyst was removed from the mixture and 10~ palladium-
carbon (80 mg) was added again. The whole was hydrogenated
for additional 24 hours at room temperature under normal
pressure. The reaction mixture was filtered to remove the
catalyst. The filtrate was concentrated to give 2-
dimethylamino-5-[1-(4-hydroxyindol-3-yl)ethyl]-2-oxazolin-
4-one (125 mg).
H-NMR (DMSO-d6) ~: 1.02-1.15(3H,m), 3.00-3.10(6H,m),
3.98(1H,m), 4.97-5.16(1H,m), 6.30-6.34(1H,m), 6.77-
6.83(2H,m), 6.95(1H,m), 9.40(1H,bs), 10.69(1H,bs).
2-dimethylamino-5-[1-(4-hydroxyindol-3-yl)ethyl]-2-
oxazolin-4-one (120 mg) was dissolved into
dimethylformamide (3 ml). Potassium carbonate (115 mg) and
indomethane (156 ~1) were added. The mixture was stirred
for 3 hours at room temperature and additional indomethane
(78 ~1) was added. The mixture was stirred for 36 hours at
room temperature. To the reaction mixture was added water
and the whole was extracted with ethyl acetate. The
extract was washed with brine and dried over magnesium
sulfate. Concentration of the ethyl acetate solution gave
a residue, which was subjected to silica gel
chromatography. El~tion with hexane-acetone (1:1) provided
2-dimethylamino-5-tl-(4-methoxyindol-3-yl)ethyl-2-oxazolin-
4-one (54 mg).
lH-NMR (CDC13) ~: 1.26(2.6H,d,J=7.0Hz),
1.45(0.4H,d,J=7.2Hz), 2.96(0.4H,s), 3.00(2.6H,s),
3.17(2.6H,s), 3.19(0.4H,s), 3.90(0.4H,s), 3.93(2.6H,s),
4.25(1H,m), 4.98(0.13H,d,J=3.4Hz), 5.24(0.87H,d,J=2.2Hz),
6.51(d,J=7.8Hz), 6.95-7.14(3H,m), 8.16(1H,bs).
2-dimethylamino-5-[1-(4-methoxyindol-3-yl)ethyl~-2-
oxazolin-4-one (50 mg) was added to methylamine (5 ml).
CA 02248784 1998-09-11
W097/49703 PCT/~97/02157- -
The mixture was refluxed for 5 hours and additional
methylamine (5 ml) was added. The whole was further
refluxed for 2.5 hours. ~he methylamine was distilled off
to give a residue, which was subjected to silica gel
chromatography. Elution with hexane-acetone (1:1) provided
the titled compound (32 mg).
IR (KBr) cm~l: 3286, 319g, 1733, 1623.
H-NMR (DMSO-d6) ~: 1.0-1.15(3H,m), 2.7-2.85(3H,m),
3.84(3H,s), 3.7-3.9(1H,m), 4.98(0.33H,d,J=2.2Hz),
5.01(0.67H,d,J=2.2Hz), 6.46(1H,m), 6.9-7.1(3H,m),
8.60(1H,bs), 10.88(1H,bs).
Example 30
5-[1-(4-chloroindol-3-yl)ethyl]-2-methylamino-2-oxazolin-4-
one
3-chloroindole (500 mg) and ethyl trans-2,3-
epoxybutyrate (472 mg) were dissolved into dichloromethane
(5 ml). To this mixture was added at -9~C over 30 minutes
titanium tetrachloride. The whole was stirred for 1 hour
at -9~C. Ethyl trans-2,3-epoxybutyrate (405 mg) and
titanium tetrachloride (405 ~1) were added. The whole was
further stirred for 1 hour at -9~C. Concentration of the
reaction mixture gave a residue, to which was added ethyl
acetate. The mixture was washed with brine and dried over
magnesium sulfate. Concentration of the solution gave a
residue, which was subjected to silica gel chromatography.
Elution with hexane-ethyl acetate (1:3) provided ethyl
(2S*,3R*)-3-(4-chloroindol-3-yl)-2-hydroxybutylate (200
mg).
lH-NMR (CDCl3) ~: 1.27(3H,d,J=7.2Hz), 1.33(3H,t,J=7.0Hz),
2.81(1H,d,J=5.2Hz), 4.25-4.40(4H,m),
4.64(1H,dd,J=5.2&2.6Hz), 7.06-7.10(2H,m), 7.24-7.29(2H,m),
8.22(lH,bs).
Sodium (100 mg) was dissolved into ethanol (5 ml).
Ethyl (2s*~3R*)-3-(4-chloroindol-3-yl)-2-hydroxybutyrate
(200 mg) and N,N'-dimethyl guanidine hydrogen bromide (143
CA 02248784 1998-09-11
W O 97/49703 ~CT/JP97102157 - -
96
mg) were added. The whole was refluxed for 16 hours. To
the reaction mixture was added water and extracted with
ethyl acetate. The extract was washed with brine and dried
over magnesium sulfate. Concentration of the solution gave
a residue, which was subjected to silica gel
chromatography. Elution with hexane-acetone (1:1) provided
the titled compound (25 mg).
H-NMR (CDC13) ~: 1.27-1.44(3H,m), 2.84-3.00(3H,m), 4.36-
4.58(lH,m), 4.99-5.11(lH,m), 5.88(lH,m), 7.00-7.30(4H,m),
8 6o-8.8o(lH~m).
Example 31
5-[1-(indol-3-yl)-2,2,2-trifluoroethyl]-2-methylamino-2-
oxazolin-4-one
Sodium ethoxide (580 mg) was added to a solution of
methyl 4,4,4-trifluoro-2-hydroxy-3-(indol-3-yl)butyrate
(980 mg) and N,N'-dimethyl guanidine hydrogen bromide (630
mg) in ethanol (2.5 ml). The mixture was refluxed for 1.5
hours and then cooled. To the cooled reaction mixture was
added ice water (20 ml). After the pH of the mixture was
adjusted to 7 using acetic acid, the whole was extracted
with ethyl acetate. The extract was washed with saturated
aqueous sodium hydrogen carbonate solution and then dried
over magnesium sulfate. Concentration of the solution
under the reduced pressure gave residue, which was
subjected to silica gel chromatography. Eluent with ethyl
acetate-hexane was collected and concentrated to provide
the titled compound (218 mg).
IR (KBr) cm~l: 3283, 1769, 1717, 1659, 1541.
lH-NMR (DMSO-d6) ~: 2.38-2.54(3H,m), 3.65-4.61(2H,m), 6.24-
6.33(1H,m), 6.94-7.98(5H,m), 11.10-11.18(1H,m).
Example 32
2-diacetylamino-5-[1-(indol-3-yl)ethyl]-2-oxazolin-4-one
2-amino-5-rl-(indol-3-yl)ethyl~-2-oxazolin-4-one (300
mg) was dissolved into tetrahydrofuran (2 ml).
CA 02248784 1998-09-11
W097/49703 PCT/~1102157
97
Triethylamine (344 ~1) and acetyl chloride (123 ~1) were
added under ice cooling. The mixture was stirred for 2
hours at 0~C. Triethylamine (344 ~1) and acetyl chloride
(123 ~1) were added again. The whole was further stirred
for 2 hours. To the reaction mixture was added water and
the whole was extracted with ethyl acetate. The extract
was washed with brine and dried over magnesium sulfate.
Concentration of the solution gave a residue, which was
subjected to silica gel chromatography. The eluent with
hexane-acetone (1:1) was collected and concentrated to
provide the titled compound (40 mg).
lH-NMR (CDC13) ~: 1.45(1.92H,d,J=7.2Hz),
1.57(1.08H,d,J=7.2Hz), 2.17(6H,s), 3.7-3.9(1H,m),
4.99(0.36H,d,J=2.4Hz), 5.07(0.64H,d,J=2.8Hz), 7.0-
7.4(4H,m), 7.6-7.7(1H,m), 8.32(1H,bs).
Example 33
2-acetylamino-5-[1-(indol-3-yl)ethyl]-2-oxazolin-4-one
2-amino-5-[1-(indol-3-yl)ethyl]-2-oxazolin-4-one (100
mg) was dissolved into pyridine (0.5 ml). Acetic anhydride
(117 ~1) was added. The mixture was stirred for 2 hours at
80~C. To the reaction mixture was added water. The
mixture was made acidic using 4N hydrochloric acid. The
whole was extracted with ethyl acetate. The extract was
washed with brine and dried over magnesium sulfate.
Concentration of the solution gave a residue, which was
subjected to the silica gel chromatography. The eluent
with hexane-acetone (1:1) was collected and concentrated to
provide the titled compound (16 mg).
IR (KBr) cm~l: 1742, 1698.
lH-~MR (CDC13) ~: 1.42(1.95H,d,J=7.2Hz),
1.62(1.05H,d,J=7.2Hz), 2.1-2.3(3H,m),
3.8B(lH,dq,J=2.2&7.2Hz), 5.00(0.35H,d,J=2.2Hz),
5.04(0.65H,d,J=2.2Hz), 7.0-7.4(5H,m), 7.67(1H,d,J=8.4Hz),
8-19(1H,bs).
_
CA 02248784 1998-09-11
W 097/49703 PCT/JP97/02157- -
98
Experimental Example 1
I~ uitro antimicrobial test:
~etermination of in vitro antimicrobial activity aqainst
Helicobacter pylori and other microorqanisms
Using 5 strains of Nelicobacterpylori, 19 other bacterial
species and 2 yeast species as test strains, the
antimicrobial activities of various compounds were
determined by the method described below (agar dilution
method). The test compound was dissolved in dimethyl
sulfoxide; this solution was 2-fold diluted step by step
with sterilized distilled water to yield test samples. Two
milliliters of each test sample was mixed with 18 ml of
Brucella agar supplemented with 7% horse blood as a medium
to prepare a determination plate. Helicobacter pylori was
lS subjected to shaking culture at 37~C in a gas pack jar
incorporating CampyPakTM (BBLR Beckton Dickinson
Microbiology Systems) for 20 hours, using Brucella broth
medium supplemented with 2.5% fetal bovine serum, to yield
a seed inoculum. The other test microorganisms were each
cultured at 37~C for 20 hours, using Brucella broth medium,
to yield respective seed inocula.
Five microliters of each seed inoculum, previously
adjusted to about 106 CFU/ml using Brucella broth medium
supplemented with 2.5% fetal bovine serum was inoculated to
each determination plate, and cultured at 37~C in a gas
pack jar incorporating CampyPakTM and water-soaked cotton
absorbent for 4 days. After cultivation, strain growth was
macroscopically examined; the minimum concentration for the
absence of strain growth was taken as the MIC value
(minimum inhibitory concentration) of the test compound.
The results are shown in Tables 2 and 3.
CA 02248784 1998-09-11
- W O 97/49703 PCT/JPg7tO2157
99
Table 2 Antimicrobial Activities of Indolmycin against
Various Microorganisms [MIC (~g/ml)]
1 Escherichia coli K12 ~100
2 Escherichia coli NIHJ JC-2 >100
3 Proteus mirabilis ATCC 21100 >100
4 Proteus vulgaris IFO 3045 >100
Proteus morganii IFO 3168 >100
6 Klebsiella pneumoniae IFO 3317 >100
7 Serratia marcescens IFO 3046 >100
8 Salmonella typhimurium IFO 12529 >100
9 Salmonella enteritidis IFO 3313 >100
Citrobater freundii IFO 12681 >100
11 Pseudomonas aeruginosa IFO 3080 >100
12 Alcaligenes faecalis IFO 13111 >100
13 Bacillus subtilis PCI 219 >100
14 Bacillus cereus IFO 3514 >100
Bacillus pumilus IFO 3813 >100
16 Bacillus megaterium IFO 12108 >100
17 Staphylococcus aureus FDA 209P >100
18 Micrococcus luteus IFO 12708 12.5
19 Micrococcus flavus IFO 3242 >100
Helicobacter pylori NCTC 11637 0.006
21 Candida albicans IFO 0583 >100
22 Saccharomyces cerevisiae IFO 0209 >100
CA 02248784 1998-09-11
- W 097/49703 PCT/JP97/02157 ~ - -
100
Table 3 Antimicrobial Activities against Various Strains
of Helicobacterpylori ( in vitro)
MIC (~q/ml)
CompoundH. pyloriH. pylori~. pyloriH. pyloriN. pylori
NCTC 11637NCTC 11916 CPY 433 TN 2 TN 58
Indolmycin 0.006 0.013 0.013 0.006 0.025
Compound of 0.025 - 0.05 0.025 0.05
Example 2
MIC was determined by agar dilution method using Brucella
agar supplemented with 7% horse blood.
As seen from Tables 2 and 3, compound (I) exhibits
very selective antimicrobial activity against the bacteria
of the genus Helicobacter, represented by Helicobacterpylori.
Experimental Example 2
In uiuo antimicrobial activity test)
After mongolian gerbils (MGS/Sea, male, 6 weeks of
age) were fasted for 24 hours, Helicobacterpylori TN2GF4 was
inoculated to the stomach at 107-08 per mongolian gerbil.
Starting at 11 days after infection, a 3, 10, 30, or 100
mg/kg suspension of the test compound in a 0.5% aqueous
solution of methyl cellulose was orally administered twice
daily (morning and evening) for 3 days. On the day after
final administration, the stomach of each infected
mongolian gerbil was excised and disrupted; a series of 10-
fold dilutions of the stomach homogenate were each
inoculated to modified Skirrow's medium supplemented with
activated charcoal and cultured under microaerophilic
conditions at 37~C for 4 days, after which clearance effect
was assessed on the basis of the presence or absence of
bacterial growth. The results are shown in Table 4.
Bacterial cell counts are expressed in mean i standard
CA 02248784 1998-09-11
W O 97/49703 PCT/JP97/02157
101
deviation after Dunnett's test against the control group.
In Table 4, ** indicates p < 0.01.
Tab}e 4
Bacterial
Dose Clearance Detection
Test Compound
(mg/kg) Rate (%) (log CFU/
gastric wall)
Control (0.5% methyl
- 0/4 ( 0) 6.36 + 0.19
cellulose solution)
Indolmycin 3 0/5 ( 0) 4.61 + 1.84
0/5 ( 0) 2.76 i 1.04**
1/4 (25) 1.96 + 0.78**
}5 100 4/5 (80) 1.48 + 0.00**
As shown in Table 4, indolmycin at 10 mg/kg or higher
reduced the mongolian gerbil gastric bacterial cell count
with dose dependency, the clearance rates achieved being
25% at 30 mg/kg and 80% at 100 mg/kg.
These findings demonstrate that the preparation of the
present invention is effective against the gastric ulcer,
gastritis and gastric cancer caused by Helicobacterpylori
infection.
Preparation Example
For use as a therapeutic agent for Helicobacterpylori
infections, the preparation of the present invention, which
contains a compound represented by formula (I) or a salt
thereof, can be produced with the following formulations:
1. Capsules
(1) Indolmycin 100 mg
(2) Lactose 90 mg
(3) Microcrystalline cellulose 70 mg
(4) Magnesium stearate 10 mg
Total 270 mg per capsule
CA 02248784 l998-09-ll
W 0 97/4g703 PCT/m97/02157
102
Components (1), (2) and (3) and a half portion of
component (4) were mixed and granulated. To these
granules, the remaining portion of component (4) was added,
and the whole mixture was packed in a gelatin capsule.
2. Tablets
(1) Indolmycin 100 mg
(2) Lactose 35 mg
(3) Corn starch 150 mg
(4) Microcrystalline cellulose 30 mg
(5) Magnesium stearate 5 mg
Total 320 mg per tablet
Components (1), (2) and (3), a two-third portion of
component (4) and a half portion of component (5) were
mixed and granulated. To these granules, the remaining
portions of components (4) and (5) were added, and the
whole mixture was tableted by compressive tableting.
Industrial ApplicabilitY
Compound (I) or a salt thereof exhibits very specific
potent antibacterial activity against the bacteria of the
genus Helicobacter represented by Nelicobacterpylori. The use of
compound (I) of the present invention or a salt thereof
therefore provides the desired effect for an anti-
Helicobacterpylori agent at doses much lower than the
effective doses of conventional antibacterial agents for
the bacteria of the genus Helicobacter ( especially Helicobacter
pylori ) .
Compound (I) or a salt thereof is effective in the
prevention or treatment of various diseases caused by
bacteria of the genus Helicobacter such as duodenal ulcer,
gastric ulcer, chronic gastritis and gastric cancer, and,
because Helicobacterpylori is a major cause of ulcer
recurrence, compound (I) or a salt thereof is also
effective in the prevention of ulcer recurrence.
Also, compound (I) or a salt thereof exhibits no
antibacterial action against gram-positive bacteria such as
CA 022487X4 1998-09-11
WO 97/49703 PCT/JP97/02157
103
those of the genera Staphylococcus and Bacillus and against
gram-negative bacteria such as those of the genera
Escherichia, Pseudomonas, Proteus, Klebsiella, Serratia, Salmonella,
Citrobacter and Alcaligenes. Compound (I) or a salt thereof is
therefore selectively effective in the prevention or
treatment of diseases caused by bacteria of the genus
Helicobacter, and can be used as a safe pharmaceutical
without adverse effects having little influence on other
bacteria and fungi.
Compound (I) or a salt thereof is stable and of low
toxicity. Accordingly, the present invention provides an
excellent anti-Helicobacterpylori agent without adverse
effects.
