U.S. patent application number 10/919475 was filed with the patent office on 2005-06-23 for novel azasugar derivative and drug containing the same as the active ingredient.
Invention is credited to Hatakeyama, Mariko, Moriyama, Hideki, Nishimura, Shinichiro, Tsukida, Takahiro, Yokota, Koichi.
Application Number | 20050137221 10/919475 |
Document ID | / |
Family ID | 29996770 |
Filed Date | 2005-06-23 |
United States Patent
Application |
20050137221 |
Kind Code |
A1 |
Tsukida, Takahiro ; et
al. |
June 23, 2005 |
Novel azasugar derivative and drug containing the same as the
active ingredient
Abstract
A novel azasugar derivative, and a drug containing the same as
an active ingredient are disclosed. The drug is useful for
treatment of keratinocyte-proliferative diseases.
Inventors: |
Tsukida, Takahiro; (Osaka,
JP) ; Moriyama, Hideki; (Sapporo-shi, JP) ;
Yokota, Koichi; (Osaka, JP) ; Hatakeyama, Mariko;
(Osaka, JP) ; Nishimura, Shinichiro; (Sapporo-shi,
JP) |
Correspondence
Address: |
WESTERMAN, HATTORI, DANIELS & ADRIAN, LLP
1250 CONNECTICUT AVENUE, NW
SUITE 700
WASHINGTON
DC
20036
US
|
Family ID: |
29996770 |
Appl. No.: |
10/919475 |
Filed: |
August 17, 2004 |
Current U.S.
Class: |
514/302 ;
514/317; 546/115; 546/219 |
Current CPC
Class: |
C07D 211/96 20130101;
A61P 17/02 20180101; A61K 31/445 20130101; C07D 491/04 20130101;
A61P 43/00 20180101; A61P 17/00 20180101 |
Class at
Publication: |
514/302 ;
514/317; 546/115; 546/219 |
International
Class: |
C07D 491/02; A61K
031/4741; A61K 031/445 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 26, 2002 |
JP |
2002-186479 |
Claims
1. A compound represented by the following formula: 80wherein Ar
represents a phenyl group which may have a substituent at the
p-position, or a pharmaceutically acceptable salt thereof:
2. A compound represented by the general formula (I): 81wherein
R.sup.1 represents a hydrogen atom, a halogen atom, a hydroxyl
group, an amino group, a C.sub.1-C.sub.8 alkyl group, a phenyl
group, a phenoxy group, a C.sub.1-C.sub.8 alkoxy group (said
C.sub.1-C.sub.8 alkoxy group may be substituted with a
C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio group),
but-2-ynyloxy group or a heteroallyloxy group, or a
pharmaceutically acceptable salt thereof.
3. A drug comprising the compound or pharmaceutically acceptable
salt thereof according to claim 1 as an active ingredient.
4. A drug comprising the compound or pharmaceutically acceptable
salt thereof according to claim 2 as an active ingredient.
5. A keratinocyte-proliferation inhibitor comprising the compound
or pharmaceutically acceptable salt thereof according to claim 1 as
an active ingredient.
6. A keratinocyte-proliferation inhibitor comprising the compound
or pharmaceutically acceptable salt thereof according to claim 2 as
an active ingredient.
7. A compound represented by the following formula (IIA) or (IIB):
82wherein Ar represents a phenyl group which may have a substituent
at the p-position, CO--R.sup.2 represents a hydroxamic acid
equivalent, and OR.sup.3 represents a hydroxyl group
equivalent.
8. A compound represented by the following formula (IIIA) or
(IIIB): 83wherein R.sup.1 represents a hydrogen atom, a halogen
atom, a hydroxyl group, an amino group, a C.sub.1-C.sub.8 alkyl
group, a phenyl group, a phenoxy group, a C.sub.1-C.sub.8 alkoxy
group (said C.sub.1-C.sub.8 alkoxy group may be substituted with a
C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio group),
but-2-ynyloxy group or a heteroallyloxy group, R.sup.4 represents a
hydrogen atom, benzyl group or an acetyl group, and R.sup.5
represents a benzyloxyamino group, a benzyloxy group, a methoxy
group or an ethoxy group.
Description
TECHNICAL FIELD
[0001] The present invention relates to a novel hydroxamic acid
compound having an azasugar structure and, more particularly, to an
azasugar derivative represented by the following general formula
(A): 1
[0002] wherein Ar represents a phenyl group which may have a
substituent at the p-position, or the following general formula
(I): 2
[0003] wherein R.sup.1 represents a hydrogen atom, a halogen atom,
a hydroxyl group, an amino group, a C.sub.1-C.sub.8 alkyl group, a
phenyl group, a phenoxy group, a C.sub.1-C.sub.8 alkoxy group (said
C.sub.1-C.sub.8 alkoxy group may be substituted with a
C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio group),
but-2-ynyloxy group or a heteroallyloxy group, or a
pharmaceutically acceptable salt thereof, and a drug containing the
same as an active ingredient.
[0004] Also the present invention is related to a compound which is
useful as an intermediate for preparation of the above compound (A)
and is represented by the following formula (IIA) or (IIB): 3
[0005] wherein Ar represents a phenyl group which may have a
substituent at the p-position, CO--R.sup.2 represents a hydroxamic
acid equivalent, and OR.sup.3 represents a hydroxyl group
equivalent.
BACKGROUND ART
[0006] Taking notice of high coordinative ability to metal atoms,
various hydroxamic acid compounds have been developed as a
metalloenzyme inhibitor. Although peptide-based compounds have
mostly been developed (see, for example, WO94/02447 and
WO90/05719), various compounds containing sulfonic acid amide in a
partial structure have recently been published and, for example,
European Patent Publication (EP1081137) discloses a compound having
the same N-sulfonylpiperidine-2-hydroxamic acid structure as that
of the compound of the present invention as an aggrecan degrading
enzyme inhibitor.
[0007] For a metalloenzyme as a target of inhibition, for example,
MMP (WO90/05716) such as collagenases, stromelysins and
gelatinases, TNF-.alpha. producing enzyme (WO 94/10990) and Fas
ligand solubilization enzyme (WO 97/09066) have become of major
interest in view of the development of medicaments.
[0008] It is known that EGF family growth factors are syntheszied
as membrane-bound proteins as precursors which are broken into a
soluble form by action of the metalloenzyme, and thus the resulting
soluble ligand binds to EGF receptors. In case of HB-EGF among
them, the solubilization process plays an important role in the
proliferation of keratinocytes (WO 01/70269). Said publication
discloses that a hydroxamic acid derivative having a
tetrahydroisoquinoline skeleton inhibits a solubilizing enzyme of
HB-EGF and is useful as a remedy for keratinocyte-proliferative
diseases.
DISCLOSURE OF THE INVENTION
[0009] An object of the present invention is to provide a novel
remedy for keratinocyte-proliferative diseases.
[0010] The present inventors have intensively studied and found
that a hydroxamic acid compound having an azasugar structure
represented by the above general formula (A) inhibits
solubilization of HB-EGF and is useful as a remedy for
keratinocyte-proliferative diseases, and thus the present invention
has been completed.
[0011] The present invention will now be described in detail.
BRIEF DESCRIPTION OF THE DRAWINGS
[0012] FIG. 1 is a graph showing hyperplasia inhibitory activity by
virtue of a compound of Example 4.
[0013] FIG. 2 is a graph showing re-epithelialization rate (%) of
keratinocytes inhibitory activity by virtue of a compound of
Example 4.
BEST MODE FOR CARRYING OUT THE INVENTION
[0014] The present invention is directed to a compound represented
by the general formula (A): 4
[0015] wherein Ar represents a phenyl group which may have a
substituent at the p-position, or a pharmaceutically acceptable
salt thereof. The substituent as used herein includes, for example,
a halogen atom, a hydroxyl group, an amino group, a C.sub.1-C.sub.8
alkyl group, a phenyl group, a phenoxy group, a C.sub.1-C.sub.8
alkoxy group (said C.sub.1-C.sub.8 alkoxy group may be substituted
with a C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio
group), but-2-ynyloxy group or a heteroallyloxy group. Among these
substituents, a methoxy group, a 2-ethoxyethoxy group,
but-2-ynyloxy group or a phenoxy group is particularly
preferable.
[0016] The compound (A) of the present invention can be prepared
from a compound (IIA) or (IIB): 5
[0017] wherein Ar represents a phenyl group which may have a
substituent at the p-position, CO--R.sup.2 represents a hydroxamic
acid equivalent, and OR.sup.3 represents a hydroxyl group
equivalent, by the method described hereinafter.
[0018] Similar to the above, the substituent includes, for example,
a halogen atom, a hydroxyl group, an amino group, a C.sub.1-C.sub.8
alkyl group, a phenyl group, a phenoxy group, a C.sub.1-C.sub.8
alkoxy group (said C.sub.1-C.sub.8 alkoxy group may be substituted
with a C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio
group), but-2-ynyloxy group or a heteroallyloxy group.
[0019] Furthermore, the hydroxamic acid equivalent as used herein
means a monovalent atomic group CONHOH itself and a functional
group which can be chemically converted into said atomic group
CONHOH by a conventional method, and R.sup.2 includes, for example,
a methoxy group, an ethoxy group, or a benzyloxyamino group. Since
said benzyloxyamino group can be easily introduced by the
condensation reaction between carboxylic acid and benzyloxyamine,
the hydroxamic acid equivalent includes a carboxyl group and
various esters which can be easily converted into a carboxyl group.
Therefore carboxylic acid protected with various carboxyl
protective groups is included and R.sup.2 includes, for example, a
benzyloxy group and a tert-butyloxy group, in addition to the
methoxy group, the ethoxy group and the benzyloxyamino group.
[0020] The hydroxyl group equivalent as used herein means a
hydroxyl group itself and a functional group which can be
chemically converted into a hydroxyl group by a conventional
method, and R.sup.3 means a hydrogen atom, and alcohol protective
group such as acetyl group, benzyl group, tert-butyl group, and
tetrahydropyranyl group.
[0021] Among the compound (IIA) or (IIB), a compound represented by
the following formula (IIIA) or (IIIB): 6
[0022] wherein R.sup.1 represents a hydrogen atom, a halogen atom,
a hydroxyl group, an amino group, a C.sub.1-C.sub.8 alkyl group, a
phenyl group, a phenoxy group, a C.sub.1-C.sub.8 alkoxy group (said
C.sub.1-C.sub.8 alkoxy group may be substituted with a
C.sub.1-C.sub.4 alkoxy group or a C.sub.1-C.sub.4 alkylthio group),
but-2-ynyloxy group or a heteroallyloxy group, R.sup.4 represents a
hydrogen atom, benzyl group or an acetyl group, and R.sup.5
represents a benzyloxyamino group, a benzyloxy group, a methoxy
group or an ethoxy group, is particularly preferably used.
[0023] The compound of the general formula (A) includes all
stereoisomers represented by the following formula: 7
[0024] or a mixture thereof.
[0025] It is considered that the configuration of a hydroxyl group
is involved in stability of the compound, and a compound (Id)
wherein hydroxyl groups at the 4- and 5-positions of an azasugar
skeleton bear R-configuration is particularly preferable.
[0026] Similarly, the compound (IIA) or (IIB) and the compound
(IIIA) or (IIIB) respectively include corresponding eight kinds of
stereoisomers or a mixture thereof. Abbreviations and symbols used
in the following descriptions mean as follows.
[0027] Ac: Acetyl group
[0028] AcOCs: Cesium acetate
[0029] Bn: Benzyl group
[0030] DEAD: Diethyl azodicarboxylate
[0031] DIEA: Diisopropylethylamine
[0032] DMF: N,N-dimethylformamide
[0033] DMP: 2,2-Dimethoxypropane
[0034] DMAP: 4-Dimethylaminopyridine
[0035] DCC: Dicyclohexylcarbodiimide
[0036] WSC: 1-Ethyl-3-(dimethylaminopropyl)carbodiimide
hydrochloride
[0037] HOBt: 1-Hydroxy-1H-benzotriazole
[0038] HPLC: High-performance liquid chromatography
[0039] THF: Tetrahydrofuran
[0040] TBAF: 1.0M THF solution of tetrabutylammonium fluoride
[0041] Pd--C: Palladium-carbon
[0042] Pd(OH) .sub.2/C: Palladium hydroxide-carbon
[0043] PPh.sub.3: Triphenylphosphine
[0044] NaOMe: Sodium methylate
[0045] NH.sub.2OBn: Benzylhydroxylamine hydrochloride
[0046] 18-Crown-6: 18-Crown-6
[0047] MsCl: Mesyl chloride
[0048] Tf.sub.2O: Trifluoromethanesulfonic anhydride
[0049] The compound (A) of the present invention can be prepared by
the method described below.
[0050] 1. Preparation of Stereoisomers (Ie) and (Ig) of Compound
(I):
[0051] For example as shown in the following scheme (Scheme 1):
8
[0052] (wherein TBDMS represents a tert-butyldimethylsilyl group
and R.sup.1 is as defined above), stereoisomers (Ie) and (Ig) can
be prepared by deprotecting a compound (IV) obtained by reacting D-
or L-threitol with a glycine derivative [see Tetrahedron Letters,
Vol. 38, No. 46, 8009-8012 (1997)] and performing the ring closure
reaction to obtain a precursor (IIIA-2), followed by deprotection
and chemical conversion of a functional group.
[0053] First, a protective group (TBDMS) of a primary hydroxyl
group of the compound (IV) [see Tetrahedron Letters, Vol. 38, No.
46, 8009-8012 (1997)] is deprotected to obtain a compound (V). The
reaction is usually conducted by adding TBAF in an inert solvent
such as THF and stirring at 0.degree. C. to room temperature for 30
minutes to 2 hours.
[0054] Then, the compound (V) is cyclized by the Mitsunobu reaction
to obtain a compound (IIIA-1). The reaction can be conducted for
example by adding triphenylphosphine and DEAD in an inert solvent
such as THF and stirring at 0.degree. C. to room temperature for 30
minutes to 4 hours. In this case, since a mixture derived from
epimerization of asymmetric carbon at the a-position of the
benzyloxycarbonyl group of the compound (IIIA-1) may appear, the
compound (IIIA-1) is subjected to the following reaction after
separation and purification by various chromatography means such as
HPLC. Alternatively, the compound (IIIA-1) may be subjected to the
following reaction in the form of a mixture and may be separated
and purified in any stage.
[0055] The resulting precursor (IIIA-1) can be converted into the
objective stereoisomers (Ie) or (Ig) by converting a
benzyloxycarbonyl group into a benzyloxyaminocarbonyl group and
performing deprotection of each group.
[0056] First, a benzyl ester moiety of the compound (IIIA-1) is
cleaved and removed by hydrogenolysis, followed by condensation
with benzylhydroxylamine to obtain a compound (IIIA-2). The
hydrogenolysis is usually conducted by optionally adding water in
ethyl acetate, lower alcohol such as methanol, or 1,4-dioxane, and
stirring in the presence of a catalyst such as 10%
palladium-carbon, 20% palladium hydroxide-carbon or platinum under
a hydrogen gas flow or pressure at room temperature to 60.degree.
C. for 1 to 5 hours. The following condensation is usually
conducted by using a condensing agent such as DCC or WSC and
stirring at 0.degree. C. to room temperature for 2 to 24 hours in
the presence of an optional auxiliary condensing agent such as
HOBt. The molar ratios of the reactants are 0.8 to 1.5 mols of
benzylhydroxylamine, 1.0 to 1.5 mol of the condensing agent and 1.0
to 1.5 mol of the auxiliary condensing agent based on 1 mol of the
carboxylic acid compound.
[0057] Then, each protective group of the compound (IIIA-2) is
cleaved and removed to obtain the objective compound. The acetyl
group, the acetonide group and the benzyl group may be cleaved and
removed by a conventional method. For example, the deacetylation
reaction is usually conducted by adding 28% NaOMe in a lower
alcohol such as methanol and stirring at 0.degree. C. to room
temperature for 1 to 6 hours. For example, the acetonide group is
cleaved and removed by optionally adding water in a solvent such as
methanol in the presence of a cation exchange resin and reacting at
room temperature to 50.degree. C. for 2 to 24 hours. The benzyl
group can be cleaved and removed by hydrogenolysis in the same
manner as described above.
[0058] In Scheme 1, a compound (IVe) as a (3S, 4S, 5R) isomer of
the compound (IV) may be used so as to obtain the stereoisomer (Ie)
and a compound (IVg) as a (3R, 4R, 5S) isomer of the compound (IV)
may be used so as to obtain the stereoisomer (Ig) (the numbering is
based on hexanoic acid). The compound (IVe) or (IVg) can be derived
from the reaction between a glycine derivative and L-threitol or
D-threitol respectively.
[0059] 2. Preparation of Stereoisomers (If) and (Ih) of Compound
(I):
[0060] For example as shown in the following scheme (Scheme 2):
9
[0061] (wherein THP represents a tetrahydropyranyl group and
R.sup.1 is as defined above), stereoisomers (If) and (Ih) can be
prepared by protecting hydroxyl groups at the 3- and 4-positions of
the stereoisomer (Ie) or (Ig) with an acetonide group and
performing steric inversion of a hydroxyl group at the 5-position
(the numbering is based on the piperidine ring).
[0062] First, protective groups (tetrahydropyranyl group and
acetonide group) of a compound (IIIA-3), which can be synthesized
in the same manner as in case of the compound (IIIA-1) shown in
Scheme 1, are cleaved and removed to obtain a compound (VI). In the
reaction, according to the method using the cation exchange resin
used in the cleavage of the above acetonide group, both protective
groups can be simultaneously cleaved and removed.
[0063] Then, acetonidation of the compound (VI) is conducted to
obtain a compound (IIIB-1). The reaction can be conducted by adding
a 5- to 15-fold amount of DMP in an inert solvent such as DMF in
the presence of an acid catalyst such as p-toluenesulfonic acid
monohydrate or camphor-10-sulfonic acid and reacting at room
temperature to 80.degree. C. for 2 to 24 hours.
[0064] A hydroxyl group of the compound (IIIB-1) is converted into
a triflate group and then steric inversion is caused by the
S.sub.N2 nucleophilic substitution reaction to obtain a compound
(IIIB-2). The reaction for preparation of a triflate compound is
usually conducted by adding trifluoromethanesulfonic anhydride in a
solvent such as methylene chloride in the presence of a base such
as triethylamine or pyridine at -60 to 0.degree. C. and stirring at
the same temperature for 30 minutes to 2 hours. In this case, the
resulting triflate compound is subjected to the following reaction
immediately after the completion of the reaction without being
purified, or can be subjected to the following reaction immediately
after purification by silica gel column chromatography. The
following S.sub.N2 nucleophilic substitution reaction can be
conducted by adding cesium acetate in a solvent such as
acetonitrile in the presence of 18-Crown-6 and stirring at
0.degree. C. to room temperature for 1 to 24 hours.
[0065] A benzyl ester moiety of the compound (IIIB-2) is cleaved
and removed by hydrogenolysis, followed by condensation with
benzylhydroxylamine to obtain a compound (IIIB-3). Subsequently,
the respective protective groups, for example, an acetyl group, an
acetonide group and a benzyl group of the compound (IIIB-3) are
cleaved and removed in any order to obtain the objective
stereoisomer (If) or (Ih). The reaction conditions are the same as
in Scheme 1.
[0066] 3. Preparation of Stereoisomers (Ia), (Ib), (Ic) and (Id) of
Compound (I):
[0067] Stereoisomers (Ia), (Ib), (Ic) and (Id) can be prepared
through the following scheme (Scheme 3): 10
[0068] (wherein R.sup.1 is as defined above)
[0069] First, an azide group of a compound (VII) [see Synthesis,
No. 9, 1305-1309 (2000)] is reduced, followed by reaction with
allylsulfonyl chloride (VIII) to obtain a compound (IX). For
example the reduction of the azide group can be conducted by
optionally adding water in ethyl acetate, lower alcohol or
1,4-dioxane, and stirring in the presence of a catalyst such as 10%
palladium-carbon, 20% palladium hydroxide-carbon or platinum under
a hydrogen gas flow or pressure at room temperature to 60.degree.
C. The following reaction with allylsulfonyl chloride (VIII) is
usually conducted in an inert solvent such as DMF in the presence
of a base such as DMAP by stirring at 0.degree. C. to room
temperature for 1 to 24 hours.
[0070] Then, a terminal acetonide group of the compound (IX) is
selectively cleaved and removed. The reaction is usually conducted
by optionally adding water in a solvent such as methanol in the
presence of a cation exchange resin and stirring at room
temperature to 50.degree. C. for 5 hours to 4 days. Alternatively,
the objective compound (X) can also be obtained by stirring in
acetonitrile at room temperature for 0.5 to 2 hours using cerium
chloride heptahydrate and oxalic acid.
[0071] Then, a primary hydroxyl group of the compound (X) is
selectively converted into a mesyl group. The reaction is conducted
by stirring in a solvent such as methylene chloride in the presence
of a base such as triethylamine or DIEA at low temperature
preferably ranging from -60 to -20.degree. C. for 30 minutes to 5
hours using 0.95 to 1.05 mol of mesyl chloride to obtain the
objective compound (XI).
[0072] Then, a compound (IIIB-4) is obtained by the intramolecular
ring closure reaction of the compound (XI). The reaction is usually
conducted in an inert solvent such as DMF in the presence of a base
such as potassium carbonate or triethylamine at room temperature to
100.degree. C., and preferably 40 to 60.degree. C. for 1 to 5 hours
to obtain the objective compound (IIIB-4).
[0073] An ester moiety of the compound (IIIB-4) is converted into
corresponding carboxylic acid by alkaline hydrolysis, which is then
condensed with benzylhydroxylamine without being purified to obtain
a compound (IIIB-5). The alkaline hydrolysis is usually conducted
by stirring in a lower alcohol such as methanol at 0 to 60.degree.
C. for 1 to 5 hours using an aqueous sodium hydroxide or lithium
hydroxide solution, and preferably an aqueous 1N solution of the
hydroxide. Subsequently, an acetonide group and a benzyl group are
cleaved and removed to obtain a compound (Ia), (Ib), (Ic) or (Id).
These reactions may be conducted in the same manner as in Scheme 1
or 2.
[0074] In Scheme 3, a compound (VIIa) as a (3R, 4S, 5S) isomer of
the compound (VII) may be used so as to obtain the stereoisomer
(Ia), a compound (VIIb) as a (3S, 4R, 5S) isomer of the compound
(VII) may be used so as to obtain the stereoisomer (Ib), a compound
(VIIc) as a (3S, 4R, 5R) isomer of the compound (VII) may be used
so as to obtain the stereoisomer (Ic), and a compound (VIId) as a
(3R, 4S, 5R) isomer of the compound (VII) may be used so as to
obtain the stereoisomer (Id) (the numbering is based on hexanoic
acid). The compound (VIIa) can be prepared from
L-gulono-1,4-lactone, the compound (VIIb) can be prepared from
L-glucono-1,5-lactone, the compound (VIIc) can be prepared from
D-gulono-1,4-lactone, and the compound (VIId) can be prepared from
D-manno-1,4-lactone [Synthesis, 9, 1305-1309 (2000)].
[0075] Futhermore, a compound of the general formula(I) wherein
R.sup.1 is a but-2-ynyloxy group, for example, can be obtained by
reacting a compound (VII) with p-benzyloxybenzenesulfonyl chloride
in place of the compound (VIII) in the same manner as in Scheme 3
to obtain a compound wherein R.sup.1 of the compound (IIIB-4) is a
benzyl group, performing hydrogenolysis of the compound to
eliminate the benzyl group, reacting with 1-bromo-2-butene in the
presence of silver oxide to obtain a compound wherein R.sup.1 of
the compound (IIIB-4) is a but-2-ynyloxy group, and the compound is
subjected to the aminolysis by the treatment with 50% hydroxylamine
aqueous solution in the presence of sodium cyanide in methanol to
afford to the objective compound.
[0076] 4. Another Method for Preparation of Stereoisomer (Id) of
compound (I):
[0077] According to the following scheme (Scheme 4) 11
[0078] (wherein R.sup.1 is as defined above), the stereoisomer (Id)
can also be prepared by protecting hydroxyl groups at the 3- and
4-positions of the stereoisomer (Ia) and performing steric
inversion of a hydroxyl group at the 5-position (the numbering is
based on the piperidine ring).
[0079] First, a compound (IIIB-7) is obtained by steric inversion
of a free hydroxyl group of a compound (IIIB-6) [one of the
compound (IIIB-5) in Scheme 3]. Specifically, said hydroxyl group
is converted into a triflate group and then the S.sub.N2
nucleophilic substitution reaction due to acetoxy ions is
conducted. The reaction conditions are the same as in Scheme 2.
[0080] Subsequently, the respective protective groups, for example,
an acetyl group, an acetonide group and a benzyl group of the
compound (IIIB-7) are cleaved and removed in any order to obtain
the objective stereoisomer (Id). The reaction conditions are the
same as in Scheme 1.
[0081] These compounds can be orally or parenterally administered
to human.
[0082] The pharmaceutical preparations for oral administration
include solid preparations such as tablets, granules, powders, fine
granules and hard capsules, and solutions such as syrups and soft
capsules. These preparations can be prepared by a conventional
method. For example, tablets, granules, powders or fine granules
are prepared by mixing the above compound or pharmaceutically
acceptable salt thereof with a conventional pharmaceutically
acceptable carrier, such as lactose, starch, crystalline cellulose,
magnesium stearate, hydroxypropyl cellulose, talc, etc., and the
hard capsules can be prepared by filling the above fine granules or
powders into suitable capsules. Besides, the syrups are prepared by
dissolving or suspending the above compound or pharmaceutically
acceptable salt thereof in an aqueous solution containing sucrose,
carboxycellulose, etc., and the soft capsules are prepared by
dissolving or suspending the compound or pharmaceutically
acceptable salt thereof in lipid excipients (for example, vegetable
oils, oily emulsion, glycol, etc.) and then filling the resultant
into soft capsules.
[0083] The pharmaceutical preparations suitable for parenteral
administration include injections and further percutaneous
preparations (ointment, lotion or cream preparation), suppositories
(for example, suppository for rectal administration, suppository
for vaginal administration), and nasal preparation (for example,
spray preparation). These preparations can be prepared by a
conventional method. For example, the injections can be prepared by
dissolving or emulsifying the compound or pharmaceutically
acceptable salt thereof in physiological saline or a lipid
excipient (for example, vegetable oil, oily emulsion, glycol, etc.)
and then filling in an ampoule or vial with sealing under sterile
condition. Ointment preparation is prepared by mixing the above
compound or pharmaceutically acceptable salt thereof with a base
such as vaseline, paraffin or glycerin, and optionally adding an
emulsifier or a preservative by a conventional method.
[0084] The dosage of the drug of the present invention may vary
depending on the dosage forms, ages, sexes, weights or conditions
of the patients, but it is usually in the range of 0.1-600 mg/kg
weight/day, and preferably 10-200 mg/kg weight/day of the active
ingredient, which is administered once a day or divided into 2 to 4
dosage units.
TEST EXAMPLE 1
[0085] 1. Test Compound
[0086] Compound a: Example 2
[0087] Compound b: Example 3
[0088] Compound c: Example 4
[0089] Compound d: Example 1
[0090] Compound e: Example 5
[0091] Compound f: Example 6
[0092] 2. Test Method
[0093] Human fibrosarcoma HT-1080 transfectant expressing fusion
protein of HB-EGF and human placental alkaline phosphatase (AP)
were used for following tests.
[0094] The cultured transfectant was treated with Trypsin-EDTA
solution, and suspended in Minimum Essential Medium without phenol
red; supplemented with 10% FCS(hereinafter referred to as MEM) at
the concentration of 1.0.times.10.sup.5 cells/ml. Each 0.2 ml of
the cell suspension was dispensed into individual wells of 96 well
microplate, and incubated overnight in a CO.sub.2 incubator at
37.degree. C. After the incubation, the culture supernatant was
removed by suction and the wells were washed with 0.2 ml/well of
phosphate buffered saline. The cells were pretreated with 0.1
ml/well of the test compound solution [dimethyl sulfoxide
(hereinafter referred to as DMSO) solution was diluted to 100 times
with MEM] for 30 min in a CO.sub.2 incubator at 37.degree. C. Then
the culture supernatant was removed by suction and cells were
treated with 0.2 ml/well of TPA solution (120 .mu.M of DMSO
solution was diluted to 60 nM with MEM) containing the same
concentration of the compound as pretreatment for 60 min in a
CO.sub.2 incubator at 37.degree. C. The cells pretreated with MEM
and incubated with only the TPA solution containing no test
compound were used as control.
[0095] After these treatments, each 0.1 ml of culture supernatant
collected from each well and 0.1 ml of MEM as background were
transferred to individual wells of new 96 well microplate and
incubated at 65.degree. C. for 10 min after plate sealing to
inactivate endogenous AP in MEM and cells. 0.1 ml of the substrate
solution (prepared by diluting an aqueous 100 mg/ml p-nitrophenyl
phosphate solution prepared previously 100 times with 0.01%
magnesium chloride/1 M diethanolamine before use) for AP was added
to each well and incubated for 120 minutes at room temperature in
the dark. The absorbance at a wavelength of 405 nm of each well was
measured using a microplate reader. The IC50 (.mu.M) values of the
test compounds were calculated by determining the concentration of
the test compounds required to exhibit 50% of the absorbance, a
difference between absorbance of the well containing only MEM as
background and absorbance of the culture supernatant in case of
adding only TPA being 100.
[0096] 3. Test Results
[0097] The test results are shown in Table 1.
1 TABLE 1 Test compounds IC50 (.mu.M) Compound a 0.61 Compound b
0.45 Compound c 0.35 Compound d 1.6 Compound e 0.34 Compound f
0.028
TEST EXAMPLE 2
[0098] 1. Test compound
[0099] Compound c (Example 4):
[0100] 2. Test Method
[0101] After backs of male BALB/c mice were shaved, 10 .mu.L of a
TPA-acetone solution (0.1 mM) was applied to the dorsal skin
surface in an area of about 1 cm.sup.2. In the medicine-treated
group, a volume of 20 .mu.L of a solution prepared by dissolving
the test compound in acetone was applied to the same area 5
minutes, 24 hours and 48 hours after TPA application. In the
non-treated group, only TPA was applied. In the control group,
acetone was applied instead of the TPA-acetone solution.
[0102] The mice were sacrificed and the skin tissues were excised
72 hours after TPA application. Then the tissues were fixed with
formalin, the vertical sections including the center of the TPA- or
acetone-treated area were prepared and embedded in paraffin
according to the conventional method. And they were stained by
hematoxylin-eosin after removing paraffin. The epidermal thickness
was measured using a microscope.
[0103] The epidermal thickness of the group treated with TPA and
the test compounds (the treated group) was compared with those of
the non-treated group and the control group, and thus hyperplasia
inhibitory activity of the test compounds was examined.
[0104] 3. Test Results
[0105] The test results are shown in FIG. 1. In FIG. 1, the numeral
in the ordinate represents the epidermal thickness (.mu.m). A bar
graph of TPA (-) represents the epidermal thickness of the control
group and a bar graph of TPA represents the epidermal thickness of
the non-treated group. The respective bar graphs of 1, 10 and 100
denote the epidermal thickness of the group wherein the test
compound is administered at a dose of 1, 10 and 100 (.mu.g/wound)
of each test compound. A significance test of the treated group
relative to the non-treated group was conducted by the Dunnett's
method. The results are indicated by the symbol * or ** (*
p<0.05, ** p<0.01). A longitudinal bar represents standard
error.
TEST EXAMPLE 3
[0106] 1. Test Compound
[0107] Compound c (Example 4):
[0108] 2. Test Method
[0109] Wound healing model was prepared in accordance with the
method described by Tsuboi, et al. (J. Dermatol., 19, 673-675
(1992)). After backs of male BALB/c mice were shaved, two
full-thickness round wounds (6 mm diameter) were prepared on the
back of each mouse using a punch biopsy instrument. In
medicine-treated group, immediately after the operation, a test
compound suspended in 0.01M phosphate-buffered saline containing
1.5% sodium carboxymethyl cellulose (vehicle) was applied to each
wound in a volume of 50 .mu.L/wound. From the following day, the
compound was repeatedly applied to each wound once a day for 7
days. In the control group, vehicle solution only was applied in
the same manner. Eight days after the operation, mice were
sacrificed and skin tissues containing wound area were removed. The
tissues were fixed with a 10% neutral buffer formalin solution for
one day and embedded in paraffin according to the conventional
method. The crosssections were made perpendicularly to the
anterior-posterior axis. Keratinocytes in the specimen were stained
by immuno histochemical staining method with an anti-keratin
antibody after removing paraffin. Measurements of wound size (mm)
and re-epithelialization of keratinocytes (mm) were performed by
using an image-analyzing software. Re-epithelialization rate (%) of
keratinocytes was calculated as follows with regard to each
animal.
Re-epithelialization rate (%) of
Keratinocytes=[(Re-epithelialization of Keratinocytes)/(Wound
Size)].times.100
[0110] 3. Test Results
[0111] The test results are shown in FIG. 2. In FIG. 2, the numeral
in the ordinate represents re-epithelialization rate (%) of
keratinocytes. Black four bar graphs denote re-epithelialization
rate (%) of keratinocytes of the treated group and the numeral at
the bottom represents a dose (.mu.g/wound) of each test compound.
The bar graph of control represents re-epithelialization rate (%)
of keratinocytes of the control group. A significance test of the
treated group relative to the control group was conducted by the
Dunnett's method. The results are indicated by the symbol **
(**p<0.01). A longitudinal bar represents standard error.
[0112] The present invention will now be described in more detail
by way of examples. The intermediates described in Examples 7 to 11
can be converted into a final compound (I) in the same manner as in
Examples 1 to 6.
EXAMPLE 1
Preparation of
(2R,3S,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl-
)-piperidine-2-carboxylic acid hydroxamide:
(1)
(3S,4'S,5'R)-3-acetoxy-3-[5'-(tert-butyldimethylsilanyloxymethyl)-2',2-
'-dimethyl-[1,3]dioxolan-4'-yl]-2-(4"-methoxybenzenesulfonylamino)-propion-
ic acid benzyl ester:
[0113] 12
[0114] A known compound (IVa) [compound of the general formula (IV)
wherein R.sup.1 is a methoxy group and an acetoxy group at the
3-position is a hydroxyl group, 20.0 g] was dissolved in pyridine
(80 mL) and acetic anhydride (40 mL) was added, and then the
mixture was stirred at room temperature for 3 hours. Methanol (20
mL) was added and, after stirring for 5 minutes, the solvent was
concentrated under reduced pressure. After the resulting residue
was dissolved in ethyl acetate (300 mL) and washed with 2.5% citric
acid (.times.2) and water, the organic layer was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl
acetate:cyclohexane=1:5.fwdarw.1:4) to obtain the titled compound
(16.2 g) as a syrup.
[0115] .sup.1H-NMR(CDCl.sub.3) .delta.: 0.07 (s), 0.10 (s),
0.85-1.0 (m), 1.2-1.4 (m), 1.44 (s), 1.96 (s), 2.07 (s), 3.5-4.0
(m), 4.1-4.2 (m), 4.40 (dd, J=1.9, 10.7 Hz), 4.48 (dd, J=2.6, 8.9
Hz), 4.8-5.3 (m), 5.56 (d, J=9.0 Hz), 6.8-6.9 (m), 7.1-7.45 (m),
7.7-7.8 (m).
(2)
(2R,3S,4'S,5'S)-3-acetoxy-3-[5'-hydroxymethyl-2',2'-dimethyl-[1,3]diox-
olan-4'-yl]-2-(4"-methoxybenzenesulfonylamino)-propionic acid
benzyl ester:
[0116] 13
[0117] The above compound (1) (16.0 g) was dissolved in THF (200
mL) and acetic acid (4.06 g) was added and also TBAF (67.2 mL) was
added under ice-cool stirring, and then the mixture was stirred at
room temperature for 1.5 hours. To the reaction solution was added
ethyl acetate (300 mL) and, after washing with water and saturated
saline, the solution was dried over magnesium sulfate and the
solvent was distilled off under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:cyclohexane
=1:3.fwdarw.1:2.fwdarw.1:1.fwdarw.3:2.fwdarw.2:1) to obtain the
titled compound (4.17 g) as a syrup.
[0118] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.22 (s, 3H), 1.36 (s, 3H),
2.07 (s, 3H), 3.5-3.7 (m, 1H), 3.75-3.9 (m, 1H), 3.86 (s, 3H),
3.9-4.2 (m, 2H), 4.47 (dd, 1H, J=2.7, 8.9 Hz), 4.90 (d, 1H, J=12.2
Hz), 5.09 (d, 1H, J=12.2 Hz), 5.14 (dd, 1H, J=2.6, 8.8 Hz), 5.55
(d, 1H, J=8.8 Hz), 6.89 (d, 2H, J=8.9 Hz), 7.2-7.4 (m, 5H), 7.73
(d, 2H, J=8.9 Hz).
(3)
(3aS,6R,7S,7aS)-7-acetoxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid benzyl
ester:
[0119] 14
[0120] The above compound (2) (4.1 g) was dissolved in THF (50 mL)
and triphenylphosphine (3.0 g) was added and, after ice cooling,
DEAD (2.0 g) was added and the mixture was stirred at room
temperature for 40 minutes. To the reaction solution was added
ethyl acetate (150 mL) and, after washing with saturated saline,
the solution was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=1:3) to obtain the titled compound (2.7 g) as a
syrup.
[0121] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.41 (s, 3H),
2.13 (s, 3H), 3.15 (dd, 1H, J=11.1, 11.9 Hz), 3.27 (dd, 1H, J=2.5,
9.3 Hz), 3.86 (s, 3H), 3.9-4.0 (m, 1H), 4.15-4.25 (m, 1H), 5.12 (d,
1H, J=12.0 Hz), 5.18 (d, 1H, J=12.0 Hz), 5.2-5.25 (m, 1H), 5.81 (t,
1H, J=2.5 Hz), 6.88 (d, 2H, J=9.0 Hz), 7.25-7.4 (m, 5H), 7.71 (d,
2H, J=9.0 Hz).
(4)
(3aS,6R,7S,7aS)-7-acetoxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
benzyloxyamide:
[0122] 15
[0123] The above compound (3) (2.7 g) was dissolved in ethyl
acetate (50 mL) and 10% Pd--C (270 mg) was added, and then the
mixture was stirred under a hydrogen atmosphere at room temperature
for one hour. The catalyst was removed by filtration and the
filtrate was concentrated under reduced pressure. The resulting
residue was dissolved in DMF (30 mL) and WSC (1.3 g) and HOBt (913
mg) were added. Subsequently, benzylhydroxylamine hydrochloride
(1.08 g) and DIEA (873 mg) were added, followed by stirring at room
temperature for 3 hours. Furthermore, WSC (299 mg), HOBt (211 mg),
benzylhydroxylamine hydrochloride (248 mg) and DIEA (202 mg) were
added, followed by stirring for 45 minutes. To the reaction
solution was added ethyl acetate (200 mL) and, after washing in
turn with 0.5N hydrochloric acid, aqueous saturated sodium
hydrogencarbonate solution and saturated saline, the solution was
dried over magnesium sulfate and the solvent was distilled off
under reduced pressure. The resulting residue was purified by
silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=1:3.fwdarw.1:2.fwd- arw.2:3) to obtain the titled
compound (2.08 g) as a syrup.
[0124] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.37 (s, 3H), 1.41 (s, 3H),
1.88 (s, 3H), 3.0-3.2 (m, 1H), 3.5-3.7 (m, 2H), 3.89 (s, 3H),
4.2-4.3 (m, 1H), 4.73 (bs, 1H), 4.89 (d, 1H, J=11 Hz), 4.96 (d, 1H,
J=11 Hz), 5.71 (t, 1H, J=2.1 Hz), 6.98 (d, 1H, J=9.0 Hz), 7.42 (s,
5H), 7.71 (d, 2H, J=9.0 Hz).
(5)
(3aS,6R,7S,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
benzyloxyamide:
[0125] 16
[0126] The above compound (4) (1.85 g) was dissolved in methanol
(25 mL) and 28% NaOMe (668 mg) was added, and then the mixture was
stirred at room temperature for 2 hours. Furthermore, 28% NaOMe
(334 mg) was added and, after stirring at room temperature for 3
hours, the reaction solution was mixed with ethyl acetate (150 mL)
and then washed with 0.5N hydrochloric acid and saturated saline.
The organic layer was dried over magnesium sulfate and the solvent
was distilled off under reduced pressure, and then the resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:cyclohexane=2:3) to obtain the titled
compound (1.36 g) as a syrup.
[0127] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.32 (s, 3H), 1.39 (s, 3H),
3.02 (dd, 1H, J=10.9, 12.8 Hz), 3.3-3.55 (m, 2H), 3.88 (s, 3H),
4.07 (dd, 1H, J=4.1, 12.8 Hz), 4.75-5.05 (m, 4H), 6.96 (d, 2H,
J=9.1 Hz), 7.3-7.5 (m, 5H), 7.80 (d, 2H, J=9.1 Hz), 9.17 (s,
1H).
(6)
(2R,3S,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid benzyloxyamide:
[0128] 17
[0129] The above compound (5) (1.1 g) was dissolved in methanol (25
mL) and a cation exchange resin (Muromac, 3.0 g) was added, and
then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=20:1) to obtain the titled compound (974 mg)
as a syrup.
[0130] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.16 (d, 1H, J=5.0 Hz),
3.4-3.7 (m, 2H), 3.81 (s, 3H), 4.27 (d, 1H, J=2.1 Hz), 4.68 (s,
2H), 4.86 (d, 1H, J=5.4 Hz), 4.94 (d, 1H, J=4.5 Hz), 5.05 (d, 1H,
J=3.8 Hz), 7.07 (d, 2H, J=8.9 Hz), 7.25-7.5 (m, 5H), 7.71 (d, 2H,
J=8.9 Hz), 11.5 (s, 1H).
(7)
(2R,3S,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid hydroxamide:
[0131] 18
[0132] The above compound (6) (316 mg) was dissolved in methanol
(25 mL) and 10% Pd--C (40 mg) was added, and then the mixture was
stirred under a hydrogen atmosphere at room temperature for 2
hours. The catalyst was removed by filtration and the filtrate was
concentrated under reduced pressure to obtain the titled compound
(242 mg) as a colorless powder.
[0133] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.25-3.7 (m, 5H), 3.83
(s, 3H), 4.34 (d, 1H, J=2.0 Hz), 4.7-5.2 (m, 3H), 7.05 (d, 2H,
J=8.9 Hz), 7.72 (d, 2H, J=8.9 Hz), 8.88 (s, 1H), 10.86 (s, 1H)
[0134] TOF-Mass: 385 (M+Na).
EXAMPLE 2
Preparation of
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl-
)-piperidine-2-carboxylic acid hydroxamide:
(1)
(2R,4'S,4"S,5'R)-(4-methoxybenzenesulfonylamino)-(2',2',2",2"-tetramet-
hyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0135] 19
[0136] A known compound
[(2R,4'S,4"S,5'R)-azide-(2',2',2",2"-tetramethyl-[-
4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester, 18.7 g]
was dissolved in ethyl acetate (180 mL) and 10% Pd--C (3.0 g) was
added, and then the mixture was stirred under a hydrogen pressure
at room temperature for 4 hours. The catalyst was removed by
filtration and the filtrate was concentrated under reduced
pressure. The resulting residue was dissolved in DMF (160 mL) and
DMAP (8.7 g) and p-methoxybenzenesulfonyl chloride (14.7 g) were
added, followed by stirring at room temperature for 12 hours. To
the reaction solution was added ethyl acetate (500 mL) and, after
washing in turn with 1N hydrochloric acid, water and saturated
saline, the organic layer was dried over magnesium sulfate and the
solvent was distilled off under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-hexane=2:3) to obtain the titled
compound (22.0 g) as a syrup.
[0137] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.37 (s, 3H), 1.39 (s, 3H),
1.42 (s, 3H), 1.43 (s, 3H), 3.56 (s, 3H), 3.86 (s, 3H), 3.85-4.3
(m, 6H), 5.36 (d, 1H, J=10.1 Hz), 6.96 (d, 2H, J=9.0 Hz), 7.76 (d,
2H, J=9.0 Hz).
(2)
(1"S,2R,4'R,5'S)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dimethyl-[1,3]dioxo-
lan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic acid methyl
ester:
[0138] 20
[0139] The above compound (1) (18.9 g) was dissolved in 90% aqueous
methanol (450 mL) and a cation exchange resin (Muromac, 16.1 g) was
added, and then the mixture was stirred at room temperature for 3
days. The insoluble material was removed by filtration and the
filtrate was concentrated under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl
acetate:n-hexane=2:3.fwdarw.chloroform:methanol=40:1) to obtain the
titled compound (10.5 g), and a starting material (6.9 g) was
recovered.
[0140] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.37 (s, 3H), 1.42 (s, 3H),
3.56 (s, 3H), 3.7-3.8 (m, 3H), 3.86 (s, 3H), 3.99 (dd, 1H, J=1.6,
9.7 Hz), 4.20 (dd, 1H, J=2.0, 8.4 Hz), 4.39 (dd, 1H, J=1.7, 8.4
Hz), 5.53 (d, 1H, J=9.8 Hz), 6.95 (d, 2H, J=9.0 Hz), 7.75 (d, 2H,
J=9.0 Hz).
(3)
(1"R,2R,4'R,5'S)-[5'-(1"-hydroxy-2"-methanesulfonyloxy-ethyl)-2',2'-di-
methyl-[1,3]dioxolan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic
acid methyl ester:
[0141] 21
[0142] The above compound (2) (3.3 g) was dissolved in methylene
chloride (50 mL) and triethylamine (1.11 mL) was added and, after
cooling to -40.degree. C., mesyl chloride (0.62 mL)/methylene
chloride (3 mL) was slowly added dropwise and the mixture was
stirred at the same temperature for 50 minutes. The reaction
solution was washed with saturated saline and the organic layer was
dried over magnesium sulfate, and then the solvent was distilled
off under reduced pressure. The resulting residue was purified by
silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=1:2.fwdarw.2:3.fwdarw.1:1) to obtain the titled
compound (1.74 g).
[0143] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.38 (s, 3H), 1.43 (s, 3H),
2.60 (d, 1H, J=8.0 Hz), 3.14 (s, 3H), 3.58 (s, 3H), 3.87 (s, 3H),
3.95-4.1 (m, 2H), 4.25-4.5 (m, 4H), 5.42 (d, 1H, J=9.6 Hz), 6.97
(d, 2H, J=9.0 Hz), 7.76 (d, 2H, J=9.0 Hz).
(4)
(3aR,4R,7S,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0144] 22
[0145] The above compound (3) (4.6 g) was dissolved in DMF (150 mL)
and potassium carbonate (1.54 g) was added, and then the mixture
was stirred at 45.degree. C. for 1.5 hours. To the reaction
solution was added ethyl acetate (300 mL) and, after washing with
water (.times.2) and saturated saline, the organic layer was dried
over magnesium sulfate and the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:n-hexane=2:3.fwdarw.1:1) to obtain the titled compound (3.1
g) as a syrup.
[0146] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.45 (s, 3H),
2.41 (d, 1H, J=4.1 Hz), 3.25 (dd, 1H, J=9.6, 12.0 Hz), 3.58 (s,
3H), 3.5-3.65 (m, 1H), 3.7-3.8 (m, 1H), 3.88 (s, 3H), 3.9-4.2 (m,
2H), 5.06 (d, 1H, J=6.1 Hz), 6.98 (d, 2H, J=9.0 Hz), 7.70 (d, 2H,
J=9.0 Hz).
(5) Preparation of
(3aS,6R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-
-2,2-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic
acid benzyloxyamide:
[0147] 23
(5-1)
(3aS,6R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid methyl
ester:
[0148] The above compound (4) (780 mg) was dissolved in methanol
(15 mL) and a cation exchange resin (Muromac, 5.0 g) was added, and
then the mixture was stirred at room temperature for 6.5 hours. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
dissolved in DMF (30 mL) and DMP (1.8 g) and p-toluenesulfonic acid
monohydrate (20 mg) were added, followed by stirring at room
temperature for 5 hours and further stirring at 50.degree. C. for
one hour. To the reaction solution was added ethyl acetate (150 mL)
and, after washing in turn with an aqueous saturated sodium
hydrogencarbonate solution, water and saturated saline, the
solution was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=2:3.fwdarw.1:1) to obtain a mixture (620 mg) of
the titled compound and the above compound (4).
(5-2)
(3aS,6R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
benzyloxyamide:
[0149] The above mixture (5-1) (620 mg) was dissolved in methanol
(30 mL) and an aqueous 1N sodium hydroxide solution (8.5 mL) was
added, and then the mixture was stirred at room temperature for 5
hours. To the reaction solution was added water (50 mL) and, after
washing with ether, the aqueous layer was acidified with an aqueous
10% citric acid solution and then extracted with ethyl acetate. The
organic layer was washed with water and saturated saline, dried
over magnesium sulfate, and then the solvent was distilled off
under reduced pressure. The resulting residue was dissolved in DMF
(30 mL) and WSC (431 mg) and HOBt (344 mg) were added.
Subsequently, benzylhydroxylamine hydrochloride (359 mg) and DIEA
(291 mg) were added, followed by stirring overnight at room
temperature. To the reaction solution was added ethyl acetate (100
mL) and, after washing in turn with an aqueous 10% citric acid
solution, an aqueous saturated sodium hydrogencarbonate solution,
water and saturated saline, the solution was dried over magnesium
sulfate and the solvent was distilled off under reduced pressure.
The resulting residue was purified by silica gel medium pressure
column chromatography (ethyl acetate:cyclohexane=2:3.fwdarw.1:1)
and the precipitated solid material was collected by filtration to
obtain the titled compound (98 mg).
[0150] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.36 (s, 3H), 1.41 (s, 3H),
2.9-3.2 (m, 2H), 3.55-3.75 (m, 2H), 3.88 (s, 3H), 4.1-4.25 (m, 2H),
4.68 (d, 1H, J=4.3 Hz), 6.98 (d, 2H, J=9.0 Hz), 7.73 (d, 2H, J=9.0
Hz), 9.1 (s, 1H).
(6) Preparation of
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulf-
onyl)-piperidine-2-carboxylic acid benzyloxyamide:
[0151] 24
[0152] The above compound (5-2) (100 mg) was dissolved in a solvent
mixture of 1,4-dioxane/methanol (5-15 mL) and a cation exchange
resin (Muromac, 2.5 g) was added, and then the mixture was stirred
at room temperature for 14 hours. The insoluble material was
removed by filtration and the filtrate was concentrated under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography
(chloroform:methanol=20:1.fwdarw.10:1) to obtain the titled
compound (88 mg).
[0153] .sup.1H-NMR(DMSO-d.sub.6+D.sub.2O) .delta.: 3.0-3.25 (m,
1H), 3.28 (dd, 1H, J=6.7, 9.4 Hz), 3.41 (t, 1H, J=11.1 Hz), 3.81
(s, 3H), 4.17 (d, 1H, J=6.8 Hz), 4.49 (d, 1H, J=10.6 Hz), 4.57 (d,
1H, J=10.6 Hz), 7.07 (d, 2H, J=8.9 Hz), 7.25-7.45 (m, 5H), 7.67 (d,
2H, J=8.9 Hz).
(7)
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid hydroxamide:
[0154] 25
[0155] The above compound (6) (88 mg) was dissolved in methanol (12
mL) and 10% Pd--C (20 mg) was added, and the mixture was stirred
under a hydrogen atmosphere at room temperature for 2 hours. After
the catalyst was removed by filtration and the filtrate was
concentrated under reduced pressure, the resulting residue was
freeze-dried to obtain the titled compound (60 mg) as a colorless
amorphous.
[0156] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.0-3.7 (m, 5H), 3.84 (s,
3H), 4.20 (d, 1H, J=5.3 Hz), 7.09 (d, 2H, J=9.0 Hz), 7.67 (d, 2H,
J=9.0 Hz), 8.82 (bs, 1H), 10.67 (s, 1H).
[0157] TOF-Mass: 385 (M+Na), 401 (M+K).
EXAMPLE 3
Preparation of
(2R,3R,4R,5S)-1-(4'-ethoxyethoxy-benzenesulfonyl)-3,4,5-tri-
hydroxy-piperidine-2-carboxylic acid hydroxamide:
(1)
(2R,4'S,4"S,5'R)-[4-(ethoxyethoxy)benzenesulfonylamino]-(2',2',2",2"-t-
etramethyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0158] 26
[0159] A known compound
[(2R,4'S,4"S,5'R)-azide-(2',2',2",2"-tetramethyl-[-
4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester, 30.6 g]
was dissolved in ethyl acetate (360 mL) and 10% Pd--C (5.3 g) was
added, and then the mixture was stirred under a hydrogen pressure
at room temperature for 4 hours. The catalyst was removed by
filtration and the filtrate was concentrated under reduced
pressure. The resulting residue was dissolved in DMF (180 mL) and
DMAP (14.2 g) was added and also p-ethoxyethoxybenzenesulfonyl
chloride (26.3 g) was added under ice-cool stirring, followed by
stirring at room temperature for 2 hours. To the reaction solution
was added ethyl acetate (500 mL) and, after washing in turn with 1N
hydrochloric acid, an aqueous saturated sodium hydrogencarbonate
solution, water and saturated saline, the organic layer was dried
over magnesium sulfate and the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:n-hexane=35:65.fwdarw.2:3) to obtain the titled compound
(35.5 g) as a syrup.
[0160] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25 (t, 3H, J=7.0 Hz),
1.37 (s, 3H), 1.39 (s, 3H), 1.43 (s, 3H), 1.44 (s, 3H), 3.57 (s,
3H), 3.61 (q, 2H, J=7.0 Hz), 3.75-3.85 (m, 2H), 3.85-4.0 (m, 2H),
4.05-4.3 (m, 6H), 5.36 (d, 1H, J=10.0 Hz), 6.99 (d, 2H, J=9.0 Hz),
7.75 (d, 2H, J=9.0 Hz).
(2) Preparation of
(1"S,2R,4'R,5'S)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dime-
thyl-[1,3]dioxolan-4'-yl]-[4-(ethoxyethoxy)benzenesulfonylamino]-acetic
acid methyl ester:
[0161] 27
[0162] The above compound (1) (35.1 g) was dissolved in 90% aqueous
methanol (800 mL) and a cation exchange resin (Muromac, 29.8 g) was
added, and then the mixture was stirred at room temperature for 4
days. The insoluble material was removed by filtration and the
filtrate was concentrated under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl
acetate:n-hexane=2:3.fwdarw.1:1.fwdarw.chloroform:methanol=25:1) to
obtain the titled compound (18.2 g), and a starting material (8.0
g) was recovered.
[0163] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25(t, 3H, J=7.0 Hz), 1.38
(s, 3H), 1.47 (s, 3H), 2.3 (bs, 1H), 2.66 (d, 1H, J=6.7 Hz),
3.5-3.7 (m, 5H), 3.7-3.85 (m, 5H), 3.97 (d, 1H, J=8.4 Hz), 4.1-4.25
(m, 3H), 4.40 (dd, 1H, J=1.7, 8.5 Hz), 5.46 (d, 1H, J=9.5 Hz), 6.99
(d, 2H, J=9.0 Hz), 7.75 (d, 2H, J=9.0 Hz).
(3)
(1"R,2R,4'R,5'S)-[4-(ethoxyethoxy)benzenesulfonylamino]-[5'-(1"-hydrox-
y-2"-methanesulfonyloxy-ethyl)-2',2'-dimethyl-[1,3)dioxolan-4'-yl]-acetic
acid methyl ester:
[0164] 28
[0165] The above compound (2) (18.2 g) was dissolved in methylene
chloride (300 mL) and triethylamine (4.25 g) was added and, after
cooling to -60.degree. C., mesyl chloride (4.59 g)/methylene
chloride (30 mL) was slowly added dropwise and the mixture was
stirred at the same temperature for 50 minutes. After the reaction
solution was washed with saturated saline and the organic layer was
dried over magnesium sulfate, the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:cyclohexane=2:3.fwdarw.1:1.fwdarw.3:2) to obtain the titled
compound (13.0 g).
[0166] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25 (t, 3H, J=7.0 Hz),
1.38 (s, 3H), 1.43 (s, 3H), 2.60 (d, 1H, J=8.0 Hz), 3.14 (s, 3H),
3.58 (q, 2H, J=7.1 Hz), 3.75-3.85 (m, 2H), 3.96 (dd, 1H, J=1.7, 9.5
Hz), 3.9-4.4 (m, 8H), 5.43 (d, 1H, J=9.5 Hz), 6.99 (d, 2H, J=9.0
Hz), 7.75 (d, 2H, J=9.0 Hz).
(4)
(3aR,4R,7S,7aR)-5-(4'-ethoxyethoxy-benzenesulfonyl)-7-hydroxy-2,2-dime-
thyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0167] 29
[0168] The above compound (3) (10.4 g) was dissolved in DMF (200
mL) and potassium carbonate (3.1 g) was added, and then the mixture
was stirred at 50.degree. C. for one hour. To the reaction solution
was added ethyl acetate (500 mL) and, after washing with water
(.times.2) and saturated saline, the organic layer was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl
acetate:n-hexane=2:3.fwdarw.1:1) to obtain the titled compound (7.3
g) as a syrup.
[0169] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25 (t, 3H, J=7.0 Hz),
1.35 (s, 3H), 1.45 (s, 3H), 3.25 (dd, 1H, J=9.7, 12.1 Hz), 3.5-3.85
(m, 9H), 3.85-4.25 (m, 4H), 5.05 (d, 1H, J=6.1 Hz), 6.99 (d, 2H,
J=9.0 Hz), 7.69 (d, 2H, J=9.0 Hz).
(5) Preparation of
(3aS,6R,7R,7aS)-5-(4'-ethoxyethoxy-benzenesulfonyl)-7-h-
ydroxy-2,2-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic
acid benzyloxyamide:
[0170] 30
(5-1)
(3aS,6R,7R,7aS)-5-(4-ethoxyethoxy-benzenesulfonyl)-7-hydroxy-2,2-dim-
ethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
methyl ester:
[0171] The above compound (4) (2.0 g) was dissolved in methanol (35
mL) and a cation exchange resin (Muromac, 8.0 g) was added, and
then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
dissolved in DMF (15 mL) and DMP (2.7 g) and p-toluenesulfonic acid
monohydrate (10 mg) were added and, after stirring overnight at
room temperature, the mixture was stirred at 50.degree. C. for one
hour. To the reaction solution was added ethyl acetate (150 mL)
and, after washing in turn with an aqueous saturated sodium
hydrogencarbonate solution, water and saturated saline, the
solution was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=2:3.fwd- arw.1:1) to obtain a mixture (817 mg) of
the titled compound and the above compound (4).
[0172] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25 (t, J=7.1Hz), 1.26 (t,
J=7.2Hz), 1.35 (s), 1.43 (s), 1.45 (s), 3.07 (d, J=5.8 Hz), 3.25
(dd, J=9.7, 12.0 Hz), 3.55 (s), 3.58 (s), 3.6-3.8 (m), 3.8-4.25
(m), 4.95 (d, J=6.6 Hz), 5.05 (d, J=6.1 Hz), 7.0 (d, 2H, J=9.0 Hz),
7.69 (d, 2H, J=9.0 Hz).
(5-2)
(3aS,6R,7R,7aS)-5-(4'-ethoxyethoxy-benzenesulfonyl)-7-hydroxy-2,2-di-
methyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
benzyloxyamide:
[0173] The above mixture (5-1) (817 mg) was dissolved in a solvent
mixture of methanol/1,4-dioxane (5-15 mL) and an aqueous 1N sodium
hydroxide solution (5 mL) was added, followed by stirring at room
temperature for one hour and 20 minutes. The reaction solution was
neutralized with an aqueous 5% citric acid solution and then
extracted with ethyl acetate. After the organic layer was washed
with water and saturated saline and dried over magnesium sulfate,
the solvent was distilled off under reduced pressure. The resulting
residue was dissolved in DMF (10 mL) and WSC (238 mg) and HOBt (190
mg) were added. Subsequently, benzylhydroxylamine hydrochloride
(198 mg) and DIEA (160 mg) were added, followed by stirring
overnight at room temperature. To the reaction solution was added
ethyl acetate (100 mL) and, after washing in turn with an aqueous
10% citric acid solution, an aqueous saturated sodium
hydrogencarbonate solution, water and saturated saline, the organic
layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. To the resulting residue was
added ether/ethyl acetate (5:1) and the precipitated solid material
was collected by filtration to obtain the titled compound (98
mg).
[0174] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.25 (t, J=7.0Hz), 1.36 (s,
3H), 1.40 (s, 3H), 2.9-3.2 (m, 2H), 3.5-3.7 (m, 4H), 3.75-3.85 (m,
2H), 4.1-4.25 (m, 4H), 4.68 (d, 1H, J=4.2 Hz), 4.88 (d, 1H, J=11.0
Hz), 4.94 (d, 1H, J=11.0 Hz), 7.01 (d, 2H, J=9.0 Hz), 7.4 (s, 5H),
7.71 (d, 2H, J=9.0 Hz), 9.10 (s, 1H).
(6) Preparation of
(2R,3R,4R,5S)-1-(4'-ethoxyethoxy-benzenesulfonyl)-3,4,5-
-trihydroxy-piperidine-2-carboxylic acid benzyloxyamide:
[0175] 31
[0176] The above compound (5-2) (5.0 g) was dissolved in methanol
(130 mL) and a cation exchange resin (Muromac, 23.4 g) was added,
and then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=20:1.fwdarw.10:1) to obtain the titled
compound (4.3 g).
[0177] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 1.1 (t, 3H, J=7.0 Hz),
3.0-3.2 (m, 1H), 3.25-3.3 (m, 1H), 3.35-3.75 (m, 7H), 4.54 (d, 1H,
J=10.6 Hz), 4.61 (d, 1H, J=10.6 Hz), 4.98 (d, 1H, J=4.7 Hz), 5.13
(d, 1H, J=4.7 Hz), 5.44 (d, 1H, J=4.6 Hz), 7.11 (d, 2H, J=8.9 Hz),
7.35 (s, 5H), 7.69 (d, 2H, J=8.9 Hz), 11.31 (s, 1H).
(7)
(2R,3R,4R,5S)-1-(4'-ethoxyethoxy-benzenesulfonyl)-3,4,5-trihydroxy-pip-
eridine-2-carboxylic acid hydroxamide:
[0178] 32
[0179] The above compound (6) (1.66 g) was dissolved in methanol
(70 mL) and 10% Pd--C (350 mg) was added, and then the mixture was
stirred under a hydrogen atmosphere at 40.degree. C. for one hour.
The catalyst was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=10:1.fwdarw.5:1) to obtain the titled compound
(815 mg) as a colorless powder.
[0180] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 1.13 (t, 3H, J=7.0 Hz),
3.0-3.3 (m, 3H), 3.50 (q, 2H, J=7.0 Hz), 3.35-3.7 (m, 2H), 3.7-3.75
(m, 2H), 4.1-4.25 (m, 3H), 4.90 (d, 1H, J=4.9 Hz), 5.07 (d, 1H,
J=4.7 Hz), 5.35 (d, 1H, J=4.5 Hz), 7.09 (d, 2H, J=8.9 Hz), 7.66 (d,
2H, J=8.9 Hz), 8.82 (d, 1H, J=1.95 Hz), 10.67 (d, 1H, J=1.95
Hz).
[0181] TOF-Mass: 443 (M+Na), 459 (M+K).
[0182] [.alpha.].sub.D 4.0.degree. [c=0.1, MeOH]
EXAMPLE 4
Preparation of
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulfonyl-
)-piperidine-2-carboxylic acid hydroxamide:
(1)
(2R,4'S,4"S,5'R)-(4-phenoxybenzenesulfonylamino)-(2',2',2",2"-tetramet-
hyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0183] 33
[0184] A known compound
[(2R,4'S,4"S,5'R)-azide-(2',2',2",2"-tetramethyl-[-
4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester, 22.4 g]
was dissolved in ethyl acetate (200 mL) and 10% Pd--C (2.2 g) was
added, and then the mixture was stirred under a hydrogen pressure
at room temperature for 4.5 hours. The catalyst was removed by
filtration and the filtrate was concentrated under-reduced
pressure. The resulting residue was dissolved in DMF (250 mL) and
DMAP (10.4 g) was added and also p-phenoxybenzenesulfonyl chloride
(20.0 g) was added, followed by stirring at room temperature for
2.5 hours. To the reaction solution was added ethyl acetate (500
mL) and, after washing in turn with 1N hydrochloric acid, water and
saturated saline, the organic layer was dried over magnesium
sulfate and the solvent was distilled off under reduced pressure.
The resulting residue was purified by silica gel medium pressure
column chromatography (ethyl acetate:n-hexane=1:3.fwdarw.1:2) to
obtain the titled compound (30.1 g) as syrup.
[0185] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.39 (s, 3H), 1.40 (s, 3H),
1.44 (s, 3H), 1.45 (s, 3H), 3.62 (s, 3H), 3.9-4.3 (m, 6H), 5.39 (d,
1H, J=10.1 Hz), 7.0-7.15 (m, 4H), 7.2-7.3 (m, 1H), 7.3-7.5 (m, 2H),
7.79 (d, 2H, J=9.1 Hz).
(2)(1"S,2R,4'R,5'S)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dimethyl-[1,3]dioxol-
an-4'-yl]-(4-phenoxybenzenesulfonylamino)-acetic acid methyl
ester:
[0186] 34
[0187] The above compound (1) (19.4 g) was dissolved in
acetonitrile (230 mL) and cerium (III) chloride heptahydrate (27.7
g) and oxalic acid (167 mg) were added, and then the mixture was
stirred at room temperature for 30 minutes. After neutralizing with
sodium carbonate, the insoluble material was removed by filtration
and washed with ethyl acetate. The filtrate and the washing were
combined and concentrated under reduced pressure, and then the
resulting residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-hexane=1:1.fwdarw.- 3:2.fwdarw.2:1)
to obtain the titled compound (8.8 g), and a starting material (7.9
g) was recovered.
[0188] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.40 (s, 3H), 2.15-2.25 (m,
1H), 2.61 (d, 1H, J=8.0 Hz), 3.63 (s, 3H), 3.7-3.85 (m, 3H), 4.00
(dd, 1H, J=1.7, 9.8 Hz), 4.23 (dd, 1H, J=2.4, 8.4 Hz), 4.43 (dd,
1H, J=1.7, 8.5 Hz), 5.46 (d, 1H, J=9.8 Hz), 7.0-7.15 (m, 4H),
7.2-7.3 (m, 1H), 7.35-7.5 (m, 2H), 7.78 (d, 2H, J=8.9 Hz).
(3) Preparation of
(1"R,2R,4'R,5'S)-[5'-(1"-hydroxy-2"-methanesulfonyloxy--
ethyl)-2',2'-dimethyl-[1,3]dioxolan-4'-yl]-(4-phenoxybenzenesulfonylamino)-
-acetic acid methyl ester:
[0189] 35
[0190] The above compound (2) (20.0 g) was dissolved in methylene
chloride (180 mL) and triethylamine (4.47 g) were added and, after
cooling to -40.degree. C., mesyl chloride (4.80 g)/methylene
chloride (40 mL) was slowly added dropwise and the mixture was
stirred at the same temperature for 40 minutes. The reaction
solution was washed with water and an aqueous saturated sodium
hydrogencarbonate solution and the organic layer was dried over
magnesium sulfate, and then the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:n-hexane=1:3.fwdarw.- 1:2.fwdarw.1:1) to obtain the titled
compound (14.1 g).
[0191] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.39 (s, 3H), 1.44 (s, 3H),
2.57 (d, 1H, J=7.9 Hz), 3.13 (s, 3H), 3.61 (s, 3H), 3.9-4.1 (m,
2H), 4.2-4.5 (m, 4H), 5.44 (d, 1H, J=9.5 Hz), 6.95-7.15 (m, 4H),
7.15-7.35 (m, 1H), 7.35-7.5 (m, 2H), 7.77 (d, 2H, J=9.0 Hz).
(4)
(3aR,4R,7S,7aR)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxybenzenesulfonyl)-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0192] 36
[0193] The above compound (3) (17.5 g) was dissolved in DMF (300
mL) and potassium carbonate (5.2 g) was added, and then the mixture
was stirred at 50.degree. C. for one hour and 20 minutes. To the
reaction solution was added ethyl acetate (500 mL) and, after
washing with water and saturated saline (.times.3), the organic
layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=2:3) to obtain the titled compound (11.4 g) as a
syrup.
[0194] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.37 (s, 3H), 1.47 (s, 3H),
2.20 (d, 1H, J=4.1 Hz), 3.28 (dd; 1H, J=9.5, 11.9 Hz), 3.55-3.6 (m,
1H), 3.74 (t, 1H, J=9.2 Hz), 3.9-4.2 (m, 3H), 5.08 (d, 1H, J=6 Hz),
7.0-7.2 (m, 4H), 7.2-7.3 (m, 1H), 7.35-7.5 (m, 2H), 7.73 (d, 2H,
J=9.0 Hz).
(5) Preparation of
(3aS,6R,7R,7aS)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxyben-
zenesulfonyl)-hexahydro-[1,3]dioxolo[4,5-c]-pyridine-6-carboxylic
acid benzyloxyamide:
[0195] 37
(5-1)
(3aS,6R,7R,7aS)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxybenzenesulfonyl)-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid methyl
ester:
[0196] The above compound (4) (11.4 g) was dissolved in methanol
(150 mL) and a cation exchange resin (Muromac, 25.0 g) was added,
and then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The precipitated solid
material was dissolved in DMF (200 mL) and DMP(20.8 g) and
p-toluenesulfonic acid monohydrate (450 mg) were added and, after
stirring overnight at room temperature, the mixture was stirred at
50.degree. C. for 2 hours. To the reaction solution was added
sodium hydrogencarbonate and the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl acetate:n-hexane=2:3)
to obtain a mixture(6.5 g) of the titled compound and the above
compound (4).
(5-2)
(3aS,6R,7R,7aS)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxybenzenesulfonyl)-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
benzyloxyamide:
[0197] The above mixture (5-1) (12.7 g) was dissolved in a solvent
mixture of methanol/1,4-dioxane (30-150 mL) and an aqueous 1N
sodium hydroxide solution (30 mL) was added, and then the mixture
was stirred at room temperature for one hour. The reaction solution
was mixed with 1N hydrochloric acid and then extracted with ethyl
acetate. After the organic layer was washed with water and
saturated saline and dried over magnesium sulfate, the solvent was
distilled off under reduced pressure. The resulting residue was
dissolved in DMF (260 mL) and WSC (10.3 g) and HOBt (8.3 g) were
added. Subsequently, benzylhydroxylamine hydrochloride (8.6 g) and
DIEA (7.0 g) were added, followed by stirring overnight at room
temperature. To the reaction solution was added ethyl acetate (500
mL) and, after washing in turn with an aqueous 10% citric acid
solution, an aqueous saturated sodium hydrogencarbonate solution,
water and saturated saline, the organic layer was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. To the resulting residue was added ether/ethyl acetate
(5:1) and the precipitated solid material was collected by
filtration to obtain the titled compound (1.28 g).
[0198] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.40 (s, 3H), 1.43 (s, 3H),
3.0-3.25 (m, 2H), 3.6-3.8 (m, 2H), 4.02 (d, 1H, J=7.9 Hz), 4.17
(dd, 1H, J=3.9, 11.4 Hz), 4.71 (d, 1H, J=5.1 Hz), 4.89 (d, 1H,
J=11.1 Hz), 4.95 (d, 1H, J=11.3 Hz), 7.0-7.15 (m, 3H), 7.25-7.3 (m,
3H), 7.35-7.5 (m, 6H), 7.74 (d, 2H, J=9.0 Hz), 9.07 (s, 1H).
(6) Preparation of
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulf-
onyl)-piperidine-2-carboxylic acid benzyloxyamide:
[0199] 38
[0200] The above compound (5-2) (1.5 g) was dissolved in methanol
(30 mL) and a cation exchange resin (Muromac, 3.4 g) was added, and
then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=20:1.fwdarw.10:1) to obtain the titled
compound (1.1 g).
[0201] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.1-3.25 (m, 1H),
3.35-3.5 (m, 3H), 4.19 (d, 1H, J=6.8 Hz), 4.53 (d, 1H, J=10.5 Hz),
4.61 (d, 1H, J=10.5 Hz), 5.01 (d, 1H, J=4.9 Hz), 5.17 (d, 1H, J=4.8
Hz), 5.49 (d, 1H, J=4.6 Hz), 7.0-7.15 (m, 4H), 7.15-7.3 (m, 1H),
7.3-7.5 (m, 7H), 7.75 (d, 2H, J=8.9 Hz), 11.32 (s, 1H)
(7)
(2R,3R,4R,5S)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid hydroxamide:
[0202] 39
[0203] The above compound (6) (1.68 g) was dissolved in methanol
(70 mL) and 10% Pd--C (350 mg) was added, and then the mixture was
stirred under a hydrogen atmosphere at 40.degree. C. for 2 hours.
The catalyst was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=20:1.fwdarw.9:1.fwdar- w.4:1) to obtain the
titled compound (951 mg) as a colorless powder.
[0204] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.0-3.7 (m, 5H), 4.20 (d,
1H, J=6.7 Hz), 4.93 (d, 1H, J=4.9 Hz), 5.11 (d, 1H, J=4.7 Hz), 5.39
(d, 1H, J=4.5 Hz), 7.0-7.25 (m, 4H), 7.25-7.35 (m, 1H), 7.4-7.55
(m, 2H), 7.73 (d, 2H, J=9.0 Hz), 8.83 (d, 1H, J=1.9 Hz), 10.66 (d,
1H, J=1.9 Hz).
[0205] TOF-Mass: 447 (M+Na), 463 (M+K).
[0206] [.alpha.].sub.D 5.9 (c=0.1, MeOH)
EXAMPLE 5
Preparation of
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulfonyl-
)-piperidine-2-carboxylic acid hydroxamide:
(1)
(2R,4'S,4"R,5'R)-(4-phenoxybenzenesulfonylamino)-(2',2',2",2"-tetramet-
hyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0207] 40
[0208] A known compound
[(2R,4'S,4"R,5'R)-azide-(2',2',2",2"-tetramethyl-[-
4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester, 30.1 g]
was dissolved in ethyl acetate (300 mL) and 10% Pd--C (4.3 g) was
added, and then the mixture was stirred under a hydrogen pressure
at 40.degree. C. for 4 hours. The catalyst was removed by
filtration and the filtrate was concentrated under reduced
pressure. The resulting residue was dissolved in DMF (390 mL) and
DMAP (18.1 g) and p-phenoxybenzenesulfonyl chloride (34.5 g) were
added, followed by stirring overnight at room temperature. To the
reaction solution was added ethyl acetate (700 mL) and, after
washing in turn with 1N hydrochloric acid, an aqueous saturated
sodium hydrogencarbonate solution, water and saturated saline, the
organic layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=1:3.fwdarw.2:3) to obtain the titled compound
(38.3 g).
[0209] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.33 (s, 3H), 1.37 (s, 3H),
1.40 (s, 3H), 1.48 (s, 3H), 3.57 (s, 3H), 3.85-4.3 (m, 6H), 5.46
(d, 1H, J=10.5 Hz), 6.95-7.1 (m, 4H), 7.15-7.3 (m, 1H), 7.35-7.5
(m, 2H), 7.7-7.85 (m, 2H).
(2)
(1"R,2R,4'R,5'S)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dimethyl-[1,3]dioxo-
lan-4'-yl]-(4-phenoxybenzenesulfonylamino)-acetic acid methyl
ester:
[0210] 41
[0211] The above compound (1) (25.8 g) was dissolved in
acetonitrile (300 mL) and cerium (III) chloride heptahydrate (36.9
g) and oxalic acid (223 mg) were added, and then the mixture was
stirred at room temperature for 70 minutes. After neutralizing with
sodium carbonate, the insoluble material was removed by filtration
and washed with ethyl acetate. The filtrate and the washing were
combined and concentrated under reduced pressure, and then the
resulting residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-hexane=2:3.fwdarw.- 3:1) to obtain
the titled compound (16.0 g), and a starting material (7.0 g) was
recovered.
[0212] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.42 (s, 3H),
2.61 (bs, 1H), 3.61 (s, 3H), 3.65-3.95 (m, 3H), 4.05-4.25 (m, 1H),
4.25-4.4 (m, 2H), 5.46 (d, 1H, J=8.6 Hz), 6.95-7.1 (m, 4H),
7.15-7.3 (m, 1H), 7.3-7.5 (m, 2H), 7.7-7.85 (m, 2H).
(3)
(1"S,2R,4'R,5'S)-[5'-(1"-hydroxy-2"-methanesulfonyloxy-ethyl)-2',2'-di-
methyl-[1,3]dioxolan-4'-yl]-(4-phenoxybenzenesulfonylamino)-acetic
acid methyl ester:
[0213] 42
[0214] The compound (25.0 g) obtained in accordance with the
procedure described in above (2) was dissolved in methylene
chloride (430 mL) and triethylamine (5.79 g) was added and, after
cooling to -40.degree. C., mesyl chloride (6.25 g)/methylene
chloride (20 mL) was slowly added dropwise and the mixture was
stirred at the same temperature for one hour. The reaction solution
was washed with water and the organic layer was dried over
magnesium sulfate, and then the solvent was distilled off under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:cyclohexane=35:65.fw- darw.2:3.fwdarw.1:1) to obtain the
titled compound (13.5 g).
[0215] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.34 (s, 3H), 1.41 (s, 3H),
2.91 (d, 1H, J=6.5 Hz), 3.13 (s, 3H), 3.61 (s, 3H), 3.85-4.0 (m,
1H), 4.05-4.28 (m, 2H), 4.28-4.4 (m, 2H), 4.54 (dd, 1H, J=2.5, 11.1
Hz), 5.47 (d, 1H, J=9.5 Hz), 6.95-7.1 (m, 4H), 7.15-7.3 (m, 1H),
7.35-7.5 (m, 2H), 7.7-7.85 (m, 2H).
(4)
(3aR,4R,7R,7aR)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxybenzenesulfonyl)-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0216] 43
[0217] The above compound (3) (13.5 g) was dissolved in DMF (320
mL) and potassium carbonate (4.0 g) was added, and then the mixture
was stirred at 45.degree. C. for one hour and 10 minutes. To the
reaction solution was added ethyl acetate (500 mL) and, after
washing with water and saturated saline (.times.2), the organic
layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:cyclohexane=2:3) to obtain the titled compound (10.3
g).
[0218] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.44 (s, 3H),
2.23 (s, 1H), 3.65 (s, 3H), 3.65-3.75 (m, 1H), 3.82 (dd, 1H, J=2.5,
9.8 Hz), 4.0-4.2 (m, 2H), 4.3-4.4 (m, 1H), 5.05 (d, 1H, J=6.3 Hz),
6.95-7.1 (m, 4H), 7.15-7.25 (m, 1H), 7.35-7.45 (m, 2H), 7.7-7.8 (m,
2H).
(5)
(3aR,4R,7R,7aR)-7-hydroxy-2,2-dimethyl-5-(4'-phenoxybenzenesulfonyl)-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0219] 44
[0220] The above compound (4) (10.2 g) was dissolved in a solvent
mixture of methanol/1,4-dioxane (30-150 mL) and an aqueous 1N
sodium hydroxide solution (55 mL) was added, and then the mixture
was stirred at room temperature for one hour and 10 minutes. The
reaction solution was neutralized with an aqueous 5% citric acid
solution and then extracted with ethyl acetate. The organic layer
was washed with water and saturated saline and dried over magnesium
sulfate, and then the solvent was distilled off under reduced
pressure. The resulting residue was dissolved in DMF (200 mL) and
WSC (5.48 g) and HOBt (3.86 g) were added. Subsequently,
benzylhydroxylamine hydrochloride (4.56 g) and a DMF (50 mL)
solution of DIEA (3.70 g) were added, followed by stirring at room
temperature for 2 hours and 30 minutes. To the reaction solution
was added ethyl acetate (500 mL) and, after washing in turn with
0.5N hydrochloric acid, an aqueous saturated sodium
hydrogencarbonate solution, water and saturated saline, the organic
layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:cyclohexane=1:1) to obtain the titled compound (8.9 g) as a
colorless powder.
[0221] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.28 (s, 3H), 1.42(s, 3H),
2.39 (s, 1H), 3.39 (d, 1H, J=14.5 Hz), 3.61 (d, 1H, J=9.1 Hz),
4.05-4.25 (m, 1H), 4.28 (s, 1H), 4.83 (d, 1H, J=11.2 Hz), 4.92 (d,
1H, J=11.2 Hz), 5.07 (d, 1H, J=5.4 Hz), 6.95-7.1 (m, 4H), 7.15-7.25
(m, 1H), 7.3-7.5 (m, 7H), 7.8-7.9 (m, 2H), 8.94 (s, 1H)
(6)
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid benzyloxyamide:
[0222] 45
[0223] The above compound (5) (8.68 g) was dissolved in methanol
(350 mL) and a cation exchange resin (Muromac, 19.0 g) was added,
and then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=1:0.fwdarw.30:1.fwdar- w.20:1.fwdarw.10:1) to
obtain the titled compound (7.35 g) as a colorless powder.
[0224] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.65-3.90 (m, 5H), 4.15
(d, 1H, J=6.1 Hz), 4.52 (d, 1H, J=10.5 Hz), 4.59 (d, 1H, J=10.5
Hz), 4.65-4.75 (m, 2H), 5.26 (d, 1H, J=4.3 Hz), 6.95-7.1 (m, 4H),
7.15-7.25 (m, 1H), 7.3-7.45 (m, 7H), 7.78 (d, 2H, J=8.8 Hz), 11.2
(s, 1H).
(7)
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-(4'-phenoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid hydroxamide:
[0225] 46
[0226] The above compound (6) (6.0 g) was dissolved in methanol
(180 mL) and 10% Pd--C (1.3 g) was added, and then the mixture was
stirred under a hydrogen atmosphere at 45.degree. C. for 2 hours
and 30 minutes. The catalyst was removed by filtration and the
filtrate was concentrated under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (chloroform:methanol=20:1.fwdarw.10- :1.fwdarw.5:1)
to obtain the titled compound (3.76 g) as a colorless powder.
[0227] Melting point: 103.5-112.degree. C.
[0228] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 3.5-3.95 (m, 5H), 4.13
(d, 1H, J=6.5 Hz), 4.55-4.7 (m, 2H), 5.16 (d, 1H, J=4.3 Hz), 7.04
(d, 2H, J=8.8 Hz), 7.14 (d, 2H, J=7.6 Hz), 7.25 (t, 1H, J=7.3 Hz),
7.4-7.55 (m, 2H), 7.78 (d, 2H, J=8.8 Hz), 8.76 (s, 1H), 10.56 (s,
1H).
[0229] TOF-Mass: 425(M+H),447(M+Na),453 (M+K).
[0230] [.alpha.].sub.D 36 (c=0.1, MeOH)
EXAMPLE 6
Preparation of
(2R,3R,4R,5R)-1-(4'-But-2'-ynyloxybenzenesulfonyl)-3,4,5-tr-
ihydroxy-piperidine-2-carboxylic acid hydroxyamide:
(1)(1"R,2R,4'R,5'S)-(4-Benzyloxybenzenesulfonylamino)-[5'-(1",2"-dihydroxy-
-ethyl)-2',2'-dimethyl[1,3]dioxolan-4'-yl]-acetic acid
methylester:
[0231] 47
[0232] A solution of compound,
(2R,4'S,4"R,5S)-azido-(2',2',2",2"-tetramet-
hyl-[4,4']bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester
(23.2g) and 10% Pd--C (2.3g) in ethyl acetate (210 mL), was
hydrogenated for 4 h at 40.degree. C., under 4 kgf/cm.sup.2
pressure. The catalyst was removed by filtration, the filtrate was
distilled off under reduced pressure. The resulting residue was
dissolved with DMF (220 mL). To the solution, DMAP (9.0 g) and
4-benzyloxybenzenesulfonyl chloride (19.9 g) were added in ice
bath, and the reaction mixture was stirred for 1.5 h at room
temperature. The mixture was diluted with ethyl acetate, and it was
washed by 1N hydrochloric acid, an aqueous saturated sodium
hydrogencarbonate solution, water and saturated saline,
successively. The organic layer was dried over magnesium sulfate
and the solvent was distilled off under reduced pressure. The
resulting residue was purified by silica gel medium pressure column
chromatography(ethyl acetate:n-hexane=1:3.fwdarw.2:3), to obtain
the compound,
(2R,4'S,4"R,5'R)-(4-Benzyloxyl-benzenesulfonylamino)-[2',2',2",2"-tetrame-
thyl-[4',4"]bis([1,3]dioxolanyl)-5'-yl]-acetic acid methyl ester,
(29.4 g) as a colorless amorphous.
[0233] To a solution of the resulting compound (43.9 g) in
acetonitrile (390 mL), cerium chloride heptahydrate (61.1 g) and
oxalic acid (369 mg) were added, and the mixture was stirred for 1
h at room temperature. Sodium carbonate was added to the mixture.
The reaction mixture was neutralized and filtered by celite pad,
and the celite pad was washed by ethyl acetate. The filtrate and
washings were combined and distilled off under reduced pressure.
The resulting residue was purified by silica gel medium pressure
column chromatography (ethyl acetate:cyclohexane=1:1.fwda- rw.3:1)
to obtain the titled compound (32.9 g) as a syrup and starting
material (12.0 g) was recovered.
[0234] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.37 (s, 3H),
1.48 (s, 3H), 2.17 (s, 1H), 2.69 (d, 1H, J=6.7 Hz), 3.55 (s, 3H),
3.7-4.0 (m, 3H), 4.0-4.25 (m, 2H), 4.25-4.45 (m, 2H), 5.18 (s, 2H),
5.45 (d, 1H, J=11.1 Hz), 7.05 (d, 2H, J=8.9 Hz), 7.25-7.5 (m, 5H),
7.78 (d, 2H, J=8.9Hz).
(2)(1"S,2R,4'R,5'S)-(4-Benzyloxybenzenesulfonylamino)-[5'-(1"-hydroxy-2"-m-
ethane-sulfonyloxy-ethyl)-2',2'-dimethyl-[1,3]dioxolan-4'-yl]-acetic
acid methyl ester:
[0235] 48
[0236] To a solution of compound above (1) (25.9 g) and
triethylamine (5.82 g) in dichloromethane (320 mL), methanesulfonyl
chloride (5.99 g) in dichloromethane (30 mL) was added at
-40.degree. C., and the mixture was stirred for 1 h at same
temperature. The reaction mixture was diluted with chloroform, the
organic layer was washed water, and dried over magnesium sulfate.
The solvent was removed under reduced pressure, and the resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-Hexane=1:2.fwdarw.2:3.fwdarw.1:1)
to obtain the titled compound (25.9 g).
[0237] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.37 (s, 3H),
1.44 (s, 3H), 2.88 (d, 1H, J=8.0 Hz), 3.16 (s, 3H), 3.59 (s, 3H),
3.94 (s, 1H), 4.05-4.25 (m, 3H), 4.25-4.45 (m, 2H), 4.56 (d, 1H,
J=11.1 Hz), 5.15 (s, 2H), 5.44 (d, 1H, J=9.6 Hz), 7.06 (d, 2H,
J=8.9 Hz), 7.3-7.5 (m, 5H), 7.78 (d, 2H, J=8.9 Hz).
(3)(3aR,4R,7R,7aR)-5-(4'-Benzyloxybenzenesulfonyl)-7-hydroxy-2,2-dimethyl--
hexahydro-[1,3]dioxolo[4,5-c)-4-carboxylic acid methyl ester:
[0238] 49
[0239] To a solution of compound above (2) (51.5 g) in DMF (500
mL), potassium carbonate (14.9 g) was added, and the reaction
mixture was stirred for 90 min at 50.degree. C. The reaction
mixture was diluted with ethyl acetate (1 L), and it was washed by
water (.times.3), brine and then dried over magnesium sulfate.
[0240] The solvent was distilled off under reduced pressure. The
resulting residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:cyclohexane=1:2) to obtain the titled
compound (30.8 g).
[0241] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.37 (s, 3H),
1.46 (s, 3H), 3.62 (s, 3H), 3.72 (d, 1H, J=13.9 Hz), 3.85 (dd, 1H,
J=9.8, 2.4 Hz), 4.05-4.2 (m, 2H), 4.39 (s, 1H), 5.08 (d, 1H, J=6.3
Hz), 5.15 (s, 2H), 7.05 (d, 2H, J=8.9 Hz), 7.3-7.5 (m, 5H), 7.77
(d, 2H, J=8.9 Hz).
(4)(3aR,4R,7R,7aR)-5-(4'-But-2'-ynyloxybenzenesulfonyl)-7-hydroxy-2,2-dime-
thyl-hexahydro-[1,3]dioxolo[4,5-c]-4-carboxylic acid methyl
ester:
[0242] 50
[0243] A solution of compound above (3) (5.8 g) and 10% Pd--C (800
mg) in ethyl acetate (200 mL), was hydrogenated for 2 h at
40.degree. C., under 3 kgf/cm.sup.2 pressure. The catalyst was
removed by filtration, the filtrate was concentrated under reduced
pressure. The residue was dissolved with acetonitrile (70 mL).
1-Bromo-2-butyne (2.4 g) and potassium carbonate (2.52 g) were
added to the solution, and the reaction mixture was stirred for 30
min at 90.degree. C. The mixture was diluted with ethyl acetate,
and it was washed by water and brine, and the organic layer was
dried over magnesium sulfate, and the solvent was removed under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl
acetate:n-hexane=1:2.fwdarw.1:1) to obtain the titled compound (4.4
g).
[0244] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.38 (s, 1H),
1.46 (s, 3H), 1.8-1.95 (m, 3H), 2.14 (t, 1H, J=1.0 Hz), 3.65 (s,
3H), 3.72 (d, 1H, J=13.9 Hz), 3.85 (dd, 1H, J=9.8, 2.5 Hz),
4.05-4.2 (m, 2H), 4.39 (s, 1H), 4.65-4.75 (m, 2H), 5.08 (d, 1H,
J=6.4 Hz), 6.95-7.1 (m, 2H), 7.7-7.85 (m, 2H).
(5)(3aR,4R,7R,7aR)-5-(4'-But-2'-ynyloxybenzenesulfonyl)-7-hydroxy-2,2-dime-
thyl-hexahydro-[1,3]dioxolo[4,5-c]-4-carboxylic acid
hydroxyamide:
[0245] 51
[0246] Aqueous 50% hydroxylamine (4 mL) was added to a solution of
compound above (4) (950 mg) and sodium cyanide (106 mg) in methanol
(20 mL), and the solution was stirred for overnight at room
temperature. The reaction mixture was concentrated under reduced
pressure, and the resulting residue was purified by silica gel
medium pressure column chromatography
(chloroform:methanol=50:1.fwdarw.30:1) to obtain the titled
compound (331 mg).
[0247] .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.: 1.25 (s, 3H),
1.30 (s, 3H), 1.75-1.9 (m, 3H), 3.67 (d, 1H, J=13.3 Hz), 3.7-3.85
(m, 2H), 4.14 (dd, 1H, J=9.7, 2.0 Hz), 4.54 (d, 1H, J=6.2 Hz),
4.75-4.9 (m, 2H), 5.26 (bs, 1H), 7.10 (d, 2H, J=8.9 Hz), 7.74 (d,
2H, J=8.9 Hz), 8.96 (s, 1H), 10.75 (s, H);
[0248] MALDI-TOF MS: 463[M+Na).sup.+, 479(M+K].sup.+.
(6)(.sup.2R,3R,4R,5R)-1-(4'-But-2'-ynyloxybenzenesulfonyl)-3,4,5-trihydrox-
y-piperidine-2-carboxylic acid hydroxyamide:
[0249] 52
[0250] To a solution of the above compound (5) (220 mg) in methanol
(12 mL), a cation exchange resin (Muromac, 2.0 g) was added, and
the mixture was stirred for overnight at 3 h. The insoluble
material was removed by the filtration, the filtrate was
concentrated under reduced pressure. The resulting mixture was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=20:1.fwdarw.5:1), and then the fraction was
lyophilized to obtain the titled compound (110 mg) as a
amorphous.
[0251] .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.: 1.8-1.9 (m,
3H), 3.55-3.68 (m, 2H), 3.72 (d, 1H, J=12.9 Hz), 3.75-3.9 (m, 2H),
4.10 (d, 1H, J=6.7 Hz), 4.62 (bs, 2H), 4.75-4.9 (m, 2H), 5.14 (bs,
1H), 7.06 (d, 2H, J=8.9 Hz), 7.74 (d, 2H, J=8.9 Hz), 8.81 (s, 1H),
10.57 (s, 1H);
[0252] MALDI-TOF MS: 423[M+Na].sup.+, 439[M+K].sup.+; Anal calcd
for (C.sub.16H.sub.20N.sub.2O.sub.8S 0.90H.sub.2O) C:46.13 H: 5.27
N: 6.72, found C:45.87 H: 4.99 N: 6.67.
EXAMPLE 7
Preparation of
(3aS,4R,7R,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-
-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
hydroxyamide:
(1)
(3S,4'S,5'R)-3-[5'-(tert-butyldimethylsilanyloxymethyl)-2',2'-dimethyl-
-[1,3]dioxolan-4'-yl]-2-(4"-methoxybenzenesulfonylamino)-3-(tetrahydropyra-
nyloxy)-propionic acid benzyl ester:
[0253] 53
[0254] A known compound (IVa) [compound of the general formula (IV)
wherein R.sup.1 is a methoxy group and an acetoxy group at the
3-position is a hydroxyl group, 21 g] was dissolved in methylene
chloride (150 mL) and dihydropyran (5.79 g) and p-toluenesulfonic
acid monohydrate (200 mg) were added, and then the mixture was
stirred at room temperature for 2.5 hours. To the reaction solution
was added chloroform (50 mL) and, after washing with saturated
saline, the solution was dried over magnesium sulfate and the
solvent was distilled off under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl
acetate:n-hexane=1:7.fwdarw.1:6.fwdarw.1:5.fwdarw.1- :4) to obtain
the titled compound (16.7 g) as a syrup.
[0255] .sup.1H-NMR(CDCl.sub.3) .delta.: 0.01-0.06 (m), 0.8-1.0 (m),
1.2-1.8 (m), 3.0-3.25 (m), 3.5-4.2 (m), 4.2-4.65 (m), 4.8-5.15 (m),
5.37 (d, J=10.7 Hz), 5.49 (d, J=8.6 Hz), 5.54 (d, J=9.9 Hz), 6.60
(d, J=10.5 Hz), 6.8-6.95 (m), 7.2-7.4 (m), 7.7-7.85 (m).
(2)
(3S,4'S,5'S)-3-[5'-hydroxymethyl-2',2'-dimethyl-[1,3]dioxolan-4'-yl]-2-
-(4"-methoxybenzenesulfonylamino)-3-(tetrahydropyranyloxy)-propionic
acid benzyl ester:
[0256] 54
[0257] The above compound (1) (16.7 g) was dissolved in THF (200
mL) and acetic acid (3.9 g) and TBAF (64.8 mL) were added, and then
the mixture was stirred overnight at room temperature. The reaction
solution was concentrated under reduced pressure and ethyl acetate
(100 mL) was added and, after washing with water, the solution was
dried over magnesium sulfate and the solvent was distilled off
under reduced pressure. The resulting residue was purified by
silica gel medium pressure column chromatography (ethyl
acetate:cyclohexane=1:1.fwdarw.2:1) to obtain the titled compound
(12.9 g) as a syrup.
[0258] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.1-1.6 (m), 3.0-3.3 (m),
3.5-4.0 (m), 4.0-4.25 (m), 4.25-4.65 (m), 4.9-5.2 (m), 5.37 (d,
J=10.8 Hz), 5.49 (d, J=8.1 Hz), 5.57 (d, J=9.8 Hz), 6.57 (d, J=10.4
Hz), 6.8-7.0 (m), 7.2-7.4 (m), 7.7-7.85 (m).
(3)
(3aS,6R,7S,7aS)-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-7-(tetrahyd-
ropyranyloxy)-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic
acid benzyl ester:
[0259] 55
[0260] The above compound (2) (12.8 g) was dissolved in THF (200
mL) and triphenylphosphine (13.9 g) and DEAD (8.95 g) were added,
and then the mixture was stirred at room temperature for 1.25
hours. To the reaction solution was added ethyl acetate (150 mL)
and, after washing with water, the solution was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl acetate:cyclohexane=1:3) to
obtain the titled compound (10.7 g) as a syrup.
[0261] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.2-1.5 (m, 6H), 1.5-2.0
(m, 6H), 3.15-3.3 (m, 2H), 3.5-3.7 (m, 1H), 3.75-4.2 (m, 3H), 3.86
(s, 3H), 4.75-4.85 (m, 1H), 4.9-4.95 (m, 1H), 5.0-5.3 (m, 3H),
6.8-6.95 (m, 2H), 7.25-7.5 (m, 5H), 7.7-7.9 (m, 2H).
(4)
(2R,3S,4R,5S)-3,4,5-trihydroxy-1-(4'-methoxybenzenesulfonyl)-piperidin-
e-2-carboxylic acid benzyl ester:
[0262] 56
[0263] The above compound (3) (10.6 g) was dissolved in methanol
(110 mL) and a cation exchange resin (Muromac, 50 g) was added, and
then the mixture was stirred overnight at room temperature. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=50:1.fwdarw.30:1.fwda- rw.10:1) to obtain the
titled compound (7.4 g) as a syrup.
[0264] .sup.1H-NMR(CDCl.sub.3) .delta.: 2.95 (t, 1H, J=11.1 Hz),
3.3-3.6 (m, 2H), 3.7-4.0 (m, 4H), 3.82 (s, 3H), 4.53 (bs, 1H),
4.9-5.2 (m, 3H), 6.82 (d, 2H, J=8.9 Hz), 7.2-7.4 (m, 5H), 7.73 (d,
2H, J=8.9 Hz).
(5)
(3aS,4R,7S,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid benzyl
ester:
[0265] 57
[0266] The above compound (4) (7.4 g) was dissolved in DMF (80 mL)
and DMP (17.6 g) and p-toluenesulfonic acid monohydrate (70 mg)
were added, and then the mixture was stirred overnight at room
temperature and further stirred at 60.degree. C. for 1.5 hours. To
the reaction solution was added ethyl acetate (150 mL) and, after
washing with an aqueous saturated sodium hydrogencarbonate solution
and saturated saline (.times.3), the organic layer was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl
acetate:cyclohexane=1:3.fwdarw.1:2.fwdarw.1:1) to obtain the titled
compound (6.3 g) as a syrup.
[0267] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.77 (s, 3H), 1.28 (s, 3H),
3.03 (d, 1H, J=6.3 Hz), 3.30 (dd, 1H, J=3.1, 12.1 Hz), 3.3-3.5 (m,
1H), 3.75-3.85 (m, 1H), 3.86 (s, 3H), 4.08 (dd, 1H, J=2.0, 5.7 Hz),
4.73 (dd, 1H, J=1.8, 6.8 Hz), 4.96 (d, 1H, J=1.8 Hz), 5.19 (d, 1H,
J=12.2 Hz), 5.26 (d, 1H, J=12.2 Hz), 6.92 (d, 2H, J=9.0 Hz), 7.4
(s, 5H), 7.78 (d, 2H, J=9.0 Hz).
(6)
(3aS,4R,7R,7aR)-7-acetoxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid benzyl
ester:
[0268] 58
[0269] The above compound (5) (4.1 g) was dissolved in methylene
chloride/pyridine (60-10 mL) and trifluoromethanesulfonic anhydride
(2.89 mL) was adding under stirring at -20.degree. C., and then the
mixture was stirred at the same temperature for 30 minutes. To the
reaction solution was added chloroform (50 mL) and, after washing
with 1N hydrochloric acid and saline, the solution was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel column
chromatography (ethyl acetate:n-hexane=1:1) to obtain a compound,
which was dissolved in acetonitrile (60 mL) and cesium acetate
(1.77 g) and 18-Crown-6 (3.05 g) were added, followed by stirring
at room temperature for 2 hours. To the reaction solution was added
ethyl acetate (100 mL) and, after washing with saturated saline,
the solution was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel column chromatography (ethyl
acetate:cyclohexane=1:4.fwdarw- .1:3) to obtain the titled compound
(791 mg) as a syrup.
[0270] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.18 (s, 3H), 1.33 (s, 3H),
1.93 (s, 3H), 3.26 (dd, 1H, J=5.9, 12.5 Hz), 3.67 (dd, 1H, J=4.3,
13.0 Hz), 3.87 (s, 3H), 4.15-4.25 (m, 1H), 4.8-4.9 (m, 2H), 5.14
(d, 1H, J=1.8 Hz), 5.23 (d, 1H, J=12.4 Hz), 5.28 (d, 1H, J=12.4
Hz), 6.93 (d, 2H, J=9.0 Hz), 7.3-7.5 (m, 5H), 7.84 (d, 2H, J=9.0
Hz).
(7)
(3aS,4R,7R,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0271] 59
[0272] The above compound (6) (730 mg) was dissolved in methanol
(10 mL) and 28% NaOMe (132 mg) was added, and then the mixture was
stirred at room temperature for one hour. Furthermore, 28% NaOMe
(132 mg) was added and, after stirring at room temperature for one
hour, the reaction solution was mixed with ethyl acetate (100 mL)
and then washed with 1N hydrochloric acid and saturated saline. The
organic layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure, and then the resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-hexane=1:1) to obtain the titled
compound (587 mg) as an amorphous.
[0273] .sup.1H-NMR(CDCl.sub.3) .delta.: 0.93 (s, 3H), 1.23 (s, 3H),
3.10 (dd, 1H, J=2.4, 11.8 Hz), 3.29 (d, 1H, J=6.6 Hz), 3.45-3.55
(m, 1H), 3.7-3.8 (m, 1H), 3.90 (s, 3H), 4.1-4.2 (m, 1H), 4.6 (d,
1H, J=2 Hz), 4.74 (dd, 1H, J=2.0, 7.3 Hz), 4.99 (s, 2H), 7.01 (d,
2H, J=9 Hz), 7.35-7.55 (m, 5H), 7.77 (d, 2H, J=9 Hz), 9.36 (s,
1H).
(8)
(3aS,4R,7R,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
hydroxyamide:
[0274] 60
[0275] The above compound (7) (278 mg) was dissolved in ethyl
acetate (15 mL) and 10% Pd--C (38 mg) was added, and then the
mixture was stirred under a hydrogen atmosphere room temperature
for 2 hours. The catalyst was removed by filtration and the
filtrate was concentrated under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (chloroform:methanol=50:1.fwdarw.30:1.fwda- rw.10:1)
and then freeze-dried to obtain the titled compound (205 mg) as a
colorless amorphous.
[0276] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 0.99 (s, 3H), 1.18 (s,
3H), 3.20 (dd, 1H, J=2.1, 10.8 Hz), 3.25-3.45 (m, 2H), 3.80 (s,
3H), 3.92 (t, 1H, J=5.3 Hz), 4.28 (d, 1H, J=6.2 Hz), 4.53 (s, 1H),
5.92 (d, 1H, J=6.6 Hz), 7.14 (d, 2H, J=9.0 Hz), 7.75 (d, 2H, J=9.0
Hz), 9.28 (s, 1H), 11.12 (s, 1H)
[0277] TOF-Mass: 425 (M+Na), 441 (M+K).
EXAMPLE 8
Preparation of
(3aR,4R,7R,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-
-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
(1)
(3aR,4R,7S,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0278] 61
[0279] The compound (1.5 g) of Example 2 (4) was dissolved in
methanol (30 mL) and an aqueous 1N sodium hydroxide solution (9.5
mL) was added, and then the mixture was stirred at room temperature
for one hour. Furthermore, an aqueous sodium hydroxide (4 mL) and
1,4-dioxane (4 mL) were added, followed by stirring for one hour.
To the reaction solution was added water (50 mL) and, after washing
with ether, the aqueous layer was acidified with an aqueous 10%
citric acid solution and extracted with ethyl acetate. The organic
layer was washed with water and saturated saline and dried over
magnesium sulfate, and then the solvent was distilled off under
reduced pressure. The resulting residue was dissolved in DMF (35
mL) and WSC (932 mg) and HOBt (744 mg) were added. Subsequently,
benzylhydroxylamine hydrochloride (776 mg) and DIEA (628 mg) were
added, followed by stirring overnight at room temperature. To the
reaction solution was added ethyl acetate (150 mL) and, after
washing in turn with an aqueous 10% citric acid solution, an
aqueous saturated sodium hydrogencarbonate solution, water and
saturated saline, the organic layer was dried over magnesium
sulfate and the solvent was distilled off under reduced pressure.
The resulting residue was purified by silica gel medium pressure
column chromatography (ethyl acetate:cyclohexane=2:3.fwdarw.1:1) to
obtain the titled compound (910 mg).
[0280] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.23 (s, 3H), 1.42 (s, 3H),
2.35 (d, 1H, J=4.6 Hz), 2.9 (dd, 1H, J=9.8, 12.1 Hz), 3.4-3.55 (m,
1H), 3.59 (dd, 1H, J=5.6, 9.6 Hz), 3.75-4.0 (m, 2H), 3.87 (s, 3H),
4.1 (d, 1H, J=14.3 Hz), 4.14 (d, 1H, J=14.3 Hz), 5.04 (d, 1H, J=5.2
Hz), 6.98 (d, 2H, J=9.0 Hz), 7.38 (s, 5H), 7.85 (d, 2H, J=9.0 Hz),
8.97 (s, 1H).
(2)
(3aR,4R,7R,7aR)-7-acetoxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0281] 62
(2-1) (3aR,4R,7S,7aR)-trifluoromethanesulfonic acid
4-benzyloxycarbamoyl-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-hexahydro-
-[1,3]dioxolo[4,5-c]pyridin-7-yl ester:
[0282] The above compound (1) (1.0 g) was dissolved in pyridine (10
mL) and trifluoromethanesulfonic anhydride (0.4 mL) was added under
stirring at -20.degree. C., and then the mixture was stirred at the
same temperature for 2 hours. To the reaction solution was added
ethyl acetate (50 mL) and, after washing in turn with dilute
hydrochloric acid, an aqueous saturated sodium hydrogencarbonate
solution and saline, the solution was dried over magnesium sulfate
and the solvent was distilled off under reduced pressure. The
resulting residue was purified by silica gel column chromatography
(ethyl acetate:n-hexane=1:3.fwdarw.1:2) to obtain the titled
compound (670 mg).
[0283] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 1.18 (s, 3H), 1.41 (s,
3H), 3.82 (s, 3H), 3.75-3.85 (m, 2H), 4.23 (d, 1H, J=5.4, 12 Hz),
4.33 (t, 1H, J=9 Hz), 4.56 (d, 1H, J=12 Hz), 4.62 (d, 1H, J=6 Hz),
4.67 (d, 1H, J=12 Hz), 5.3-5.35 (m, 1H), 7.1-7.2 (m, 2H), 7.3-7.5
(m, 5H), 7.75-7.8 (m, 2H), 11.67 (s, 1H).
(2-2)
(3aR,4R,7R,7aR)-7-acetoxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0284] The above compound (2-1) (670 mg) was dissolved in
acetonitrile (30 mL) and cesium acetate (0.5 g) and 18-Crown-6 (1.4
g) were added, and then the mixture was stirred at room temperature
for 16 hours, To the reaction solution was added ethyl acetate (100
mL) and, after washing in turn with dilute hydrochloric acid, an
aqueous saturated sodium hydrogencarbonate solution and saturated
saline, the solution was dried over magnesium sulfate and the
solvent was distilled off under reduced pressure. The resulting
residue was purified by silica gel column chromatography (ethyl
acetate:n-hexane=1:3.fwdarw.1:1) to obtain the titled compound (161
mg).
[0285] .sup.1H-NMR(DMSO-d.sub.6) .delta.: 1.26 (s, 3H), 1.33 (s,
3H), 3.82 (s, 3H), 3.75-3.95 (m, 3H), 4.24 (dd, 1H, J=3, 10.2 Hz),
4.63 (d, 1H, J=10.8 Hz), 4.72 (d, 1H, J=10.8 Hz), 4.75 (d, 1H,
J=6.6 Hz), 5.35-5.4 (m, 1H), 7.1-7.2 (m, 2H), 7.3-7.45 (m, 5H),
7.7-7.75 (m, 2H), 11.55 (s, 1H).
(3)
(3aR,4R,7R,7aR)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0286] 63
[0287] The above compound (2-2) (100 mg) was dissolved in methanol
(2 mL) and 28% sodium methoxide (18 mg) was added, and then the
mixture was stirred at room temperature for 2 hours. After
neutralizing with a cation exchange resin, the insoluble material
was removed by filtration and the filtrate was concentrated under
reduced pressure. The resulting residue was purified by silica gel
column chromatography (ethyl
acetate:n-hexane=1:2.fwdarw.1:1.fwdarw.2:1) to obtain the titled
compound (69 mg).
[0288] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.43 (s, 3H), 1.53 (s, 3H),
3.36 (d, 1H, J=14.4 Hz), 3.5-3.65 (m, 1H), 3.86 (s, 3H), 3.8-3.9
(m, 2H), 4.1-4.2 (m, 1H), 4.27 (bs, 1H), 4.86 (d, 1H, J=11.4 Hz),
4.93 (d, 1H, J=11.4 Hz), 5.08 (bs, 1H), 6.9-7.0 (m, 2H), 7.3-7.4
(m, 5H), 7.8-7.9 (m, 2H), 8.88 (s, 1H).
EXAMPLE 9
Preparation of
(3aS,4R,7S,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-
-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
methyl ester:
(1)
(2R,4'R,4"S,5'S)-(4-methoxybenzenesulfonylamino)-(2',2',2",2"-tetramet-
hyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0289] 64
[0290] Using a known compound
[(2R,4'S,4"R,5'S)-azide-(2',2',2",2"-tetrame-
thyl-[4',4")bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester,
11.7 g], the titled compound (11.9 g) was obtained as a colorless
solid in the same manner as in Example 2 (1).
[0291] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.28 (s, 3H), 1.33 (s, 3H),
1.36 (s, 3H), 1.46 (s, 3H), 3.55 (s, 3H), 3.87 (s, 3H), 3.8-4.0 (m,
3H), 4.05-4.25 (m, 3H), 5.58 (d, 1H, J=7.9 Hz), 6.95 (d, 2H, J=8.9
Hz), 7.78 (d, 2H, J=8.9 Hz).
(2)
(1"S,2R,4'S,5'R)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dimethyl-[1,3]dioxo-
lan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic acid methyl
ester:
[0292] 65
[0293] Using the above compound (1) (14.7 g), the titled compound
(7.1 g) was obtained in the same manner as in Example 2 (2), and a
starting material (4.3 g) was recovered.
[0294] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.32 (s, 3H), 1.36 (s, 3H),
1.9 (bs, 1H), 3.54 (s, 3H), 3.65-3.8 (m, 2H), 3.86 (s, 3H),
3.8-4.05 (m, 1H), 4.12 (d, 1H, J=5.8 Hz), 4.25-4.34 (m, 1H), 6.97
(d, 2H, J=9.0 Hz), 7.79 (d, 2H, J=9.0 Hz).
(3)
(1"R,2R,4'S,5'R)-[5'-(1"-hydroxy-2"-methanesulfonyloxy-ethyl)-2',2'-di-
methyl-[1,3]dioxolan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic
acid methyl ester:
[0295] 66
[0296] Using the above compound (2) (7.06 g), the titled compound
(2.6 g) was obtained in the same manner as in Example 2 (3).
[0297] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.30 (s, 3H), 1.35 (s, 3H),
3.12 (s, 3H), 3.57 (s, 3H), 3.87 (s, 3H), 3.9-4.0 (m, 2H), 4.0-4.2
(m, 1H), 4.2-4.35 (m, 2H), 4.45-4.55 (m, 1H), 6.98 (d, 2H, J=9.0
Hz), 7.78 (d, 2H, J=9.0 Hz).
(4)
(3aS,4R,7S,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0298] 67
[0299] Using the above compound (3) (2.47 g), the titled compound
(1.01 g) was obtained as a syrup in the same manner as in Example 2
(4).
[0300] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.44 (s, 3H), 1.46 (s, 3H),
2.375 (d, 1H, J=1.4 Hz), 3.23 (dd, 1H, J=5.1, 14.7 Hz), 3.46 (dd,
1H, J=4.5, 9.8 Hz), 3.75 (s, 3H), 3.8-3.9 (m, 1H), 3.88 (s, 3H),
4.1-4.2 (m, 1H), 4.35-4.4 (m, 1H), 4.41 (d, 1H, J=8.7 Hz), 6.99 (d,
2H, J=9.0 Hz), 7.80 (d, 2H, J=9.0 Hz).
EXAMPLE 10
Preparation of
(3aS,4R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-
-dimethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
(1)
(2S,4'S,4"R,5'R)-acetoxy-(2',2',2",2"-tetramethyl-[4',4"]bis[[1,3]diox-
olanyl]-5'-yl)-acetic acid methyl ester:
[0301] 68
[0302] A compound
[(2R,4'S,4"R,5'R)-hydroxy-(2',2',2",2"-tetramethyl-[4',4-
"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl ester, see
Helvetica Chimica Acta, Vol.71, pages 609-618 (1988), 30 g] was
dissolved in methylene chloride/pyridine (350-39 mL) and
trifluoromethanesulfonic anhydride/methylene chloride (20-10 mL)
was added under stirring at -20.degree. C., and then the mixture
was stirred at the same temperature for one hour. To the reaction
solution was added chloroform (50 mL) and, after washing with
water, the solution was dried over magnesium sulfate and the
solvent was distilled off under reduced pressure. The resulting
residue was purified by silica gel column chromatography (ethyl
acetate:n-hexane=1:3) to obtain a compound, which was dissolved in
acetonitrile (130 mL) and cesium acetate (4.89 g) and 18-Crown-6
(8.45 g) were added, followed by stirring at room temperature for
one hour. To the reaction solution was added ethyl acetate (200 mL)
and, after washing with water and saturated saline, the solution
was dried over magnesium sulfate and the solvent was distilled off
under reduced pressure. The resulting residue was purified by
silica gel medium pressure column chromatography (ethyl
acetate:n-hexane=1:3) to obtain the titled compound (9.0 g) as a
syrup.
[0303] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.41 (s, 3H), 1.46 (s, 3H),
1.47 (s, 3H), 1.48 (s, 3H), 2.24 (s, 3H), 3.83 (s, 3H), 3.93 (dd,
1H, J=6.7, 8.4 Hz), 4.05-4.2 (m, 2H), 4.5-4.65 (m, 1H), 4.50 (dd,
1H, J=5.3, 7.8 Hz), 5.16 (d, 1H, J=3.1 Hz).
(2)
(2S,4'S,4"R,5'R)-hydroxy-(2',2',2",2"-tetramethyl-[4',4"]bis[[1,3]diox-
olanyl]-5'-yl)-acetic acid methyl ester:
[0304] 69
[0305] The compound above (1) (9.0 g) was dissolved in methanol (45
mL) and 28% NaOMe (2.61 mL) was added, and then the mixture was
stirred at room temperature for 3 hours and 40 minutes. The
reaction solution was mixed with ethyl acetate (200 mL) and then
washed with 1N hydrochloric acid and saturated saline. The organic
layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure, and then the resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:n-hexane=1:3.fwdarw.3:7) to obtain
the titled compound (3.0 g) as a syrup.
[0306] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.41 (s, 3H), 1.42 (s, 3H),
1.43 (s, 3H), 2.0-2.15 (m, 1H), 3.7-4.0 (m, 3H), 3.81 (s, 3H),
4.0-4.15 (m, 1H), 4.38 (dd, 1H, J=5.3, 7.5 Hz).
(3)
(2R,4'R,4"R,5'S)-azide-(2',2',2",2"-tetramethyl-[4',4"]bis[[1,3]dioxol-
anyl]-5'-yl)-acetic acid methyl ester:
[0307] 70
[0308] The above compound (2) (3.0 g) was dissolved in methylene
chloride/pyridine (50-4 mL) and trifluoromethanesulfonic anhydride
(2.0 mL) was added under stirring at -30.degree. C., and then the
mixture was stirred at the same temperature for one hour. The
reaction solution was mixed with chloroform (50 mL) and then washed
with 1N hydrochloric acid and saturated saline. The organic layer
was dried over magnesium sulfate and the solvent was distilled off
under reduced pressure. Then, the resulting residue was dissolved
in DMF (30 mL) and sodium azide (1.21 g) were added, followed by
stirring at room temperature for 2.5 hours. To the reaction
solution was added ethyl acetate (50 mL) and, after washing with 1N
hydrochloric acid and saturated saline, the solution was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl acetate:n-hexane=1:3) to
obtain the titled compound (2.89 g) as a syrup.
[0309] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.37 (s, 3H), 1.43 (s, 6H),
1.44 (s, 3H), 3.83 (s, 3H), 3.85-3.95 (m, 1H), 4.0-4.23 (m, 3H),
4.40 (dd, 1H, J=4.8, 7.2 Hz).
(4)
(2R,4'R,4"R,5'S)-(4-methoxybenzenesulfonylamino)-(2',2',2",2"-tetramet-
hyl-[4',4"]bis[[1,3]dioxolanyl]-5'-yl)-acetic acid methyl
ester:
[0310] 71
[0311] Using the above compound (3) (18.8 g), the titled compound
(27.4 g) was obtained as a colorless solid in the same manner as in
Example 2 (1).
[0312] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.32 (s, 3H), 1.378 (s,
3H), 1.382 (s, 3H), 1.44 (s, 3H), 3.50 (s, 3H), 3.87 (s, 3H),
3.85-4.10 (m, 5H), 4.20-4.25 (m, 1H), 5.38 (d, 1H, J=9.5 Hz), 6.97
(d, 2H, J=9.0 Hz), 7.76 (d, 2H, J=9.0 Hz).
(5)
(1"R,2R,4'S,5'R)-[5'-(1",2"-dihydroxy-ethyl)-2',2'-dimethyl-[1,3]dioxo-
lan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic acid methyl
ester:
[0313] 72
[0314] Using the above compound (4) (19.8 g), the titled compound
(11.6 g) was obtained in the same manner as in Example 4 (2), and a
starting material (2.6 g) was recovered.
[0315] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.38 (s, 3H),
3.49 (s, 3H), 3.7-3.8 (m, 3H), 3.86 (s, 3H), 3.85-4.25 (m, 3H),
6.97 (d, 2H, J=9.0 Hz), 7.76 (d, 2H, J=9.0 Hz).
(6)
(1"S,2R,4'S,5'R)-[5'-(1"-hydroxy-2"-methanesulfonyloxy-ethyl)-2',2'-di-
methyl-[1,3]dioxolan-4'-yl]-(4-methoxybenzenesulfonylamino)-acetic
acid methyl ester:
[0316] 73
[0317] Using the above compound (5) (11.1 g), the titled compound
(7.6 g) was obtained in the same manner as in Example 2 (3).
[0318] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.39 (s, 3H),
2.71 (d, 1H, J=8.0 Hz), 3.13 (s, 3H), 3.51 (s, 3H), 3.87 (s, 3H),
3.9-4.4 (m, 6H), 5.60 (d, 1H, J=9.9 Hz), 6.98 (d, 2H, J=9.0 Hz),
7.77 (d, 2H, J=9.0 Hz).
(7)
(3aS,4R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid methyl
ester:
[0319] 74
[0320] Using the above compound (6) (7.5 g), the titled compound
(6.05 g) was obtained as a syrup in the same manner as in Example 2
(4).
[0321] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.44 (s, 6H), 2.74 (d, 1H,
J=7.2 Hz), 3.26 (dd, 1H, J=2.8, 14.9 Hz), 3.45-3.75 (m, 3H), 3.80
(s, 3H), 3.88 (s, 3H), 3.95-4.1 (m, 1H), 4.33 (d, 1H, J=8.9 Hz),
6.99 (d, 2H, J=9.0 Hz), 7.84 (d, 2H, J=9.0 Hz).
(8)
(3aS,4R,7R,7aS)-7-hydroxy-5-(4'-methoxybenzenesulfonyl)-2,2-dimethyl-h-
exahydro-[1,3]dioxolo[4,5-c]pyridine-4-carboxylic acid
benzyloxyamide:
[0322] 75
[0323] Using the above compound (7) (350 mg), the titled compound
(114 mg) was obtained in the same manner as in Example 8 (1).
[0324] .sup.1H-NMR(CDCl.sub.3) .delta.: 1.35 (s, 3H), 1.41 (s, 3H),
2.57 (d, 1H, J=7.7 Hz), 3.32 (dd, 1H, J=7.3, 10.2 Hz), 3.48 (d, 1H,
J=2.8 Hz), 3.87 (s, 3H), 3.85 (dd, 1H, J=8.8, 10.3 Hz), 3.9-4.0 (m,
1H), 4.27 (d, 1H, J=8.5 Hz), 4.93 (d, 1H, J=11.0 Hz), 4.99 (d, 1H,
J=10.9 Hz), 6.99 (d, 2H, J=9.0 Hz), 7.3-7.5 (m, 5H), 7.84 (d, 2H,
J=9.0 Hz), 8.86 (s, 1H).
EXAMPLE 11
Preparation of
(2R,3R,4R,5R)-3-benzyloxy-1-(4'-but-2'-ynyloxybenzenesulfon-
yl)-4,5-dihydroxypiperidine-2-carboxylic acid hydroxamide:
(1)
(3aR,6R,7R,7aS)-7-hydroxy-5-(4'-benzyloxybenzenesulfonyl)-2,2-dimethyl-
-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid methyl
ester:
[0325] 76
[0326] The compound (22 g) of Example 6 (3) was dissolved in
methanol (250 mL) and a cation exchange resin (Muromac, 44 g) was
added, and then the mixture was stirred overnight at room
temperature. The insoluble material was removed by filtration and
the filtrate was concentrated under reduced pressure. The resulting
residue was purified by silica gel medium pressure column
chromatography (ethyl acetate:cyclohexane=5:1.fwdarw.ethy- l
acetate:methanol=10:1) to obtain a triol compound (12.1 g). This
compound was dissolved in DMF (120 mL) and DMP (28.8 g) and
p-toluenesulfonic acid monohydrate (300 mg) were added, followed by
stirring at room temperature for 67 hours. To the reaction solution
was added ethyl acetate (500 mL) and, after washing with water
(.times.2) and saturated saline, the organic layer was dried over
magnesium sulfate and the solvent was distilled off under reduced
pressure. The resulting residue was purified by silica gel medium
pressure column chromatography (ethyl
acetate:cyclohexane=1:2.fwdarw.1:1.fwdarw.2:1) to obtain the titled
compound (9.4 g) as a syrup.
[0327] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.10 (s, 3H),
1.27 (s, 3H), 2.96 (d, 1H, J=3.7 Hz), 3.68 (dd, 1H, J=14.7, 2.6
Hz), 3.75 (s, 3H), 3.9-4.05 (m, 2H), 4.20 (dd, 1H, J=6.7, 5.2 Hz),
4.25-4.35 (m, 1H), 4.38 (d, 1H, J=4.0 Hz), 5.15 (s, 2H), 7.06 (d,
2H, J=9.0 Hz), 7.3-7.55 (m, 2H), 7.81 (d, 2H, J=9.0 Hz).
(2)
(3aR,6R,7R,7aS)-7-hydroxy-5-(4'-but-2'-ynyloxybenzenesulfonyl)-2,2-dim-
ethyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
methyl ester:
[0328] 77
[0329] The above compound (1) (3.0 g) was dissolved in ethyl
acetate (20 mL) and 10% Pd--C (200 mg) was added, and then the
mixture was stirred under a hydrogen atmosphere at 40.degree. C.
for 1.5 hours. After the catalyst was removed by filtration and the
filtrate was concentrated under reduced pressure, the resulting
residue was dissolved in acetonitrile (25 mL) and 1-bromo-2-butyne
(1.25 g) and silver oxide (1.3 g) were added, followed by stirring
under a nitrogen atmosphere at 90.degree. C. for 30 minutes. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography (ethyl
acetate:cyclohexane=1:3.fwdarw.1:2) to obtain the titled compound
(2.1 g) as a syrup.
[0330] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.12 (s, 3H),
1.28 (s, 3H), 1.88 (s, 3H), 3.00 (d, 1H, J=3.7 Hz), 3.69 (dd, 1H,
J=14.8, 2.5 Hz), 3.78 (s, 3H), 3.9-4.1 (m, 2H), 4.21 (dd, 1H,
J=6.8, 5.1 Hz), 4.25-4.35 (m, 1H), 4.38 (d, 1H, J=4.0 Hz), 4.73 (q,
1H, J=2.2 Hz), 7.06 (d, 2H, J=8.9 Hz), 7.82 (d, 2H, J=8.9 Hz).
(3)(3aR,6R,7R,7aS)-7-benzyloxy-5-(4'-but-2'-ynyloxybenzenesulfonyl)-2,2-di-
methyl-hexahydro-[1,3]dioxolo[4,5-c]pyridine-6-carboxylic acid
methyl ester:
[0331] 78
[0332] The above compound (2) (932 mg) was dissolved in methylene
chloride (18 mL) and benzyl bromide (1.09 g) and silver oxide(1.47
g) were added, and then the mixture was stirred under a nitrogen
atmosphere at room temperature for 6 days. The insoluble material
was removed by filtration and the filtrate was concentrated under
reduced pressure. The resulting residue was purified by silica gel
medium pressure column chromatography (ethyl acetate:n-hexane=1:3)
to obtain the titled compound (802 mg) as a syrup.
[0333] .sup.1H-NMR (500 MHz, CDCl.sub.3) .delta.: 1.25 (s, 3H),
1.32 (s, 3H), 1.88 (s, 3H), 3.60 (s, 3H), 3.68 (dd, 1H, J=14.3, 2.8
Hz), 3.80 (t, 1H, J=5.8 Hz), 3.96 (dd, 1H, J=14.3, 1.6 Hz), 4.3-4.4
(m, 2H), 4.63 (d, 1H, J=12.4 Hz), 4.66 (d, 1H, J=12.4 Hz), 4.7-4.75
(m, 2H), 4.76 (d, 1H, J=5.8 Hz), 7.00 (d, 2H, J=9.0 Hz), 7.25-7.5
(m, 5H), 7.76 (d, 2H, J=9.0 Hz).
(4)
(2R,3R,4R,5R)-3-benzyloxy-1-(4'-but-2'-ynyloxybenzenesulfonyl)-4,5-dih-
ydroxypiperidine-2-carboxylic acid hydroxamide:
[0334] 79
[0335] The above compound (3) (300 mg) was dissolved in methanol
(15 mL) and an aqueous 1N sodium hydroxide solution (1.7 mL) was
added, and then the mixture was stirred at room temperature for 4
hours. To the reaction solution was added water (50 mL) and, after
washing with ether, the aqueous layer was acidified with an aqueous
5% citric acid solution and then extracted with chloroform. The
organic layer was washed with saturated saline and dried over
magnesium sulfate, and then solvent was distilled off under reduced
pressure. The resulting residue was dissolved in DMF (10 mL) and
WSC (72 mg) and HOBt (51 mg) were added. Subsequently,
O-(tert-butyldimethylsilyl)hydroxylamine (64 mg) was added,
followed by stirring overnight at room temperature. To the reaction
solution was added ethyl acetate (50 mL) and, after washing in turn
with an aqueous 5% citric acid solution, an aqueous saturated
sodium hydrogencarbonate and saturated saline (.times.2), the
organic layer was dried over magnesium sulfate and the solvent was
distilled off under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=50:1.fwdarw.20:1) to obtain a hydroxamic acid
compound.
[0336] The resulting hydroxamic acid compound was dissolved in
methanol (15 mL) and a cation exchange resin (Muromac, 2 g) was
added, and then the mixture was stirred overnight at room
temperature and further stirred at 50.degree. C. for 3 hours. The
insoluble material was removed by filtration and the filtrate was
concentrated under reduced pressure. The resulting residue was
purified by silica gel medium pressure column chromatography
(chloroform:methanol=50:1.fwdarw.20:1) and then freeze-dried to
obtain the titled compound (58 mg) as an amorphous.
[0337] .sup.1H-NMR (500 MHz, DMSO-d.sub.6) .delta.: 1.83 (t, 3H,
J=2.2 Hz), 3.53 (dd, 1H, J=9.8, 6.5 Hz), 3.55-3.75 (m, 2H), 3.79
(bs, 1H), 4.0-4.1 (m, 1H), 4.44 (d, 1H, J=6.5 Hz), 4.46 (d, 1H,
J=11.6 Hz), 4.65-4.8 (m, 3H), 4.83 (d, 2H, J=2.3 Hz), 7.04 (d, 2H,
J=9.0 Hz), 7.2-7.4 (m, 5H), 8.91 (s, 1H), 10.91 (s, 1H);
[0338] MALDI-TOF MS: 513[M+Na].sup.+, 529[M+K].sup.+.
EXAMPLE 12
[0339] Preparation of Tablets:
[0340] Tablets each containing 100 mg of
(2R,3R,4R,5R)-3,4,5-trihydroxy-1--
(4'-phenoxybenzenesulfonyl)-piperidine-2-carboxylic acid
hydroxamide (compound e) of Example 5 are obtained by the following
procedure.
2 Ingredients Amount Active principle (Compound e) 100 Parts by
weight Cornstarch 46 Parts by weight Microcrystalline cellulose 98
Parts by weight Hydroxypropyl cellulose 2 Parts by weight Magnesium
stearate 4 Parts by weight
[0341] [Procedure]
[0342] An active principle, cornstarch and microcrystalline
cellulose are mixed and to the mixture is added hydroxypropyl
cellulose dissolved in 50 parts by weight of water, followed by
sufficient kneading. The kneaded mixture is passed through a sieve
to granulate, dried, mixed with magnesium stearate and then
compressed into tablets of 250 mg each.
EXAMPLE 13
[0343] Preparation of Granules:
[0344] Granules containing
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-(4'-phenoxyben-
zenesulfonyl)-piperidine-2-carboxylic acid hydroxamide (compound e)
of Example 5 are obtained by the following procedure.
[0345] [Formulation]
3 Ingredients Amount Active principle (Compound e) 200 Parts by
weight Lactose 185 Parts by weight Cornstarch 109 Parts by weight
Hydroxypropyl cellulose 6 Parts by weight
[0346] [Procedure]
[0347] An active principle, lactose and cornstarch are mixed and to
the mixture is added hydroxypropyl cellulose dissolved in 120 parts
by weight of water, followed by sufficient kneading. The kneaded
mixture is passed through a 20 mesh sieve to granulate, dried and
then size-adjusted to obtain granules containing 200 mg of an
active principle per 500 mg of granule.
EXAMPLE 14
[0348] Preparation of Capsules:
[0349] Capsules each containing 100 mg of
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-
-(4'-phenoxybenzenesulfonyl)-piperidine-2-carboxylic acid
hydroxamide (compound e) of Example 5 are obtained by the following
procedure.
[0350] [Formulation]
4 Ingredients Amount Active principle (Compound e) 100 Parts by
weight Lactose 35 Parts by weight Cornstarch 60 Parts by weight
Magnesium stearate 5 Parts by weight
[0351] [Procedure]
[0352] The above ingredients are well mixed and 200 mg each of the
powder mixture is encapsulated to obtain capsules.
EXAMPLE 15
[0353] Preparation of Injections:
[0354] A mixture of 0.5 parts by weight of
(2R,3R,4R,5R)-3,4,5-trihydroxy--
1-(4'-phenoxybenzenesulfonyl)-piperidine-2-carboxylic acid
hydroxamide (compound e) of Example 5 and 5 parts by weight of
sorbitol are dissolved in distilled water for injection to obtain
100 parts by weight of the aqueous solution. The aqueous solution
is filtered through a membrane filter. 5 g each of the filtrate is
poured into an ample substituted by nitrogen gas. The ample are
sealed and then sterilized by heating at 120.degree. C. for 15
minutes to obtain injection containing 25 mg of the compound (e)
per ample.
EXAMPLE 16
[0355] Preparation of Ointments:
[0356] 1.0 parts by weight of
(2R,3R,4R,5R)-3,4,5-trihydroxy-1-(4'-phenoxy-
benzenesulfonyl)-piperidine-2-carboxylic acid hydroxamide (compound
e) and 0.1 parts by weight of butyl paraben are dispersed in 5.0
parts by weight of light liquid paraffin. The mixture is milled in
a mortar and sieved through a 200 mesh screen. This product is
mixed with 5.0 parts by weight of liquid paraffin and the mixture
is mixed with 88.9 parts by weight of gelled hydrocarbon warmed at
about 60.degree. C. to take a homogenous dispersion whereupon oily
ointment is obtained.
[0357] Industrial Applicability
[0358] The drugs of the present invention suppress the release of
HB-EGF from cell membrane stimulated with
12-O-tetradecanoylphorbol-13-acetate (hereinafter referred to as
TPA) (see, Test example 1). They also suppress not only epidermal
hyperplasia in mice induced by the application of TPA on their
back, but re-epithelialization of keratinocytes mouse skin wound
model (see, Test Examples 2 and 3). Severe side effects were not
noted in these Test Examples using mice. Thus, the medicine
according to the present invention is useful as a
keratinocyte-proliferation inhibitor.
* * * * *