U.S. patent application number 14/586403 was filed with the patent office on 2015-04-30 for sterol derivative.
This patent application is currently assigned to KYOWA HAKKO KIRIN CO., LTD. The applicant listed for this patent is KYOWA HAKKO KIRIN CO. LTD.. Invention is credited to Tsutomu AGATSUMA, Kazuhiro HIBINO, Kyoichiro IIDA, Hideyuki ONODERA, Setsuya SASHO, Kenji UCHIDA.
Application Number | 20150119585 14/586403 |
Document ID | / |
Family ID | 43499154 |
Filed Date | 2015-04-30 |
United States Patent
Application |
20150119585 |
Kind Code |
A1 |
UCHIDA; Kenji ; et
al. |
April 30, 2015 |
STEROL DERIVATIVE
Abstract
The present invention provides a sterol derivative or a
pharmaceutically acceptable salt thereof having an activity to
promote proliferation of neural stem cells. Namely, the present
invention provides a sterol derivative represented by the general
formula (I) (wherein Y represents optionally substituted lower
alkyl or the like; X.sup.a and X.sup.b are the same or different,
and represent a bond or the like; R.sup.1, R.sup.2, R.sup.3,
R.sup.4, R.sup.7 and R.sup.8 are the same or different, and
represent a hydrogen atom or the like; R.sup.5 and R.sup.6 are the
same or different, and represent a hydrogen atom or the like;
R.sup.9 represents a hydrogen atom or the like; R.sup.10 and
R.sup.11 together represent a bond or the like; and R.sup.12
represents a hydrogen atom or the like) or a pharmaceutically
acceptable salt thereof. ##STR00001##
Inventors: |
UCHIDA; Kenji; (Shizuoka,
JP) ; AGATSUMA; Tsutomu; (Tokyo, JP) ; HIBINO;
Kazuhiro; (Tokyo, JP) ; SASHO; Setsuya;
(Shizuoka, JP) ; IIDA; Kyoichiro; (Kanagawa,
JP) ; ONODERA; Hideyuki; (Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
KYOWA HAKKO KIRIN CO. LTD. |
Tokyo |
|
JP |
|
|
Assignee: |
KYOWA HAKKO KIRIN CO., LTD
Tokyo
JP
|
Family ID: |
43499154 |
Appl. No.: |
14/586403 |
Filed: |
December 30, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13386542 |
Mar 15, 2012 |
8952003 |
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PCT/JP2010/062297 |
Jul 22, 2010 |
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14586403 |
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Current U.S.
Class: |
549/276 |
Current CPC
Class: |
A61P 21/02 20180101;
A61P 25/00 20180101; A61K 31/585 20130101; C07J 9/00 20130101; C07J
75/005 20130101; A61P 25/28 20180101; A61P 25/18 20180101; A61P
43/00 20180101; A61P 25/24 20180101; A61P 9/10 20180101; A61P 9/00
20180101; A61P 25/08 20180101; C07J 73/003 20130101; A61P 25/16
20180101; A61P 25/22 20180101; C07J 73/008 20130101 |
Class at
Publication: |
549/276 |
International
Class: |
C07J 73/00 20060101
C07J073/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 24, 2009 |
JP |
2009-172886 |
Claims
1-16. (canceled)
17. A method for preparing a compound represented by the following
formula (I-b): ##STR00150## comprising a step of converting a
compound represented by the following formula (IV) to a compound
represented by the following formula (V): ##STR00151## wherein,
R.sup.22 represents a protective group for a hydroxy group, Y
represents an optionally substituted lower alkyl or an optionally
substituted lower alkenyl, and represents a single bond or a double
bond.
18. The method according to claim 17, wherein the formula (I-b) is
the following formula (P) or (Q). ##STR00152##
19. The method according to claim 17, wherein the protective group
for a hydroxy group is trimethyl silyl, tetrahydropyranyl or
benzoyl.
20. The method according to claim 18, wherein the protective group
for a hydroxy group is trimethyl silyl, tetrahydropyranyl or
benzoyl.
Description
TECHNICAL FIELD
[0001] The present invention relates to a sterol derivative having
an activity to promote proliferation of neural stem cells.
BACKGROUND ART
[0002] Neurodegenerative diseases are diseases in which cerebral
and peripheral nerve cells are damaged by a hereditary factor, an
environmental factor, an aging factor and the like. Specifically,
they include Parkinson's disease, Alzheimer's disease, triplet
repeat disease, amyotrophic lateral sclerosis, polyneuropathy,
spinal cord injury, cerebrovascular disorders and the like.
[0003] Although a general therapeutic method for these
neurodegenerative diseases is a method in which neurotransmitters
lost by the injury of nerve cells are supplemented, the diseases
for which the therapeutic method is effective are limited to
Parkinson's disease, Alzheimer's disease and the like at present.
Additionally, the progress of nerve cell death cannot be stopped by
the neurotransmitter supplementation method.
[0004] Regenerative medicine which regenerates the central nervous
system has been investigated from the viewpoint of transplantation,
as a therapeutic method for positively recovering the function of
dopaminergic neurons which had lost by Parkinson's disease.
However, the regenerative medicine has not been generally used due
to various problems caused by the use of aborted fetal brain.
Additionally, studies have also been conducted on a therapeutic
method in which neural stem cells obtained from a fetal brain or ES
cells obtained from a human fertilized eggs are mass-cultured in
vitro and differentiated into a neuron of interest to use it for
transplantation (Stem Cells, 2006, vol. 24, p. 1583-1593; The
Journal of Neuroscience, 2005, vol. 25, p. 4694-4705). However, its
clinical applications are not in progress since the techniques for
accurately differentiating them into the desired neuron have not
been established yet; teratomas are formed by undifferentiated
cells; and there are problems caused by the use of fetal neural
stem cells or human ES cells. Accordingly, a technique in which
adult-derived neural stem cells are cultured in vitro and used for
transplantation is regarded as a promising technique and search for
factors which efficiently accelerate proliferation of neural stem
cells is expected (Nature Reviews Neuroscience, 2006, vol. 7, p.
395-406).
[0005] As a low molecular compound which promotes proliferation of
neural stem cells, for example, Salvianolic acid B (JP2006-76948),
hedgehog signal agonists (Journal of Biology, 2002, vol. 1, p. 10),
selective serotonin reuptake inhibitors (Science, 2003, vol. 301,
p. 805-809; Proceedings of the National Academy of Science of the
United States of America, 2006, vol. 103, p.8233-8238),
metabotropic glutamate receptor antagonists (Biochemical and
Biophysical Research Communications, 2004, vol. 315, p. 493-496),
PPAR.gamma. agonists (The Journal of Biological Chemistry, 2006,
vol. 281, p. 12673-12681), NMDA agonists (Journal of Cell Science,
2007, vol. 120, p. 1358-1370) and the like have been reported.
[0006] On the other hand, as a sterol derivative, the following
compounds (1) to (4) are known (see Non-Patent Literatures 1 to
4).
##STR00002##
PRIOR ART DOCUMENTS
Non-Patent Literature
[0007] Non-Patent Literature 1: Heterocycles, 1985, vol. 23, p.
1607-1610 [0008] Non-Patent Literature 2: Tetrahedron Letters,
1979, vol. 20, p. 3119-3122 [0009] Non-Patent Literature 3: CAS
REGISTRY Database, Registry Number: 6048-74-4 [0010] Non-Patent
Literature 4: Agricultural and Biological Chemistry, 1964, vol. 28,
p. 788-795
DISCLOSURE OF THE INVENTION
Problems to be Solved by the Invention
[0011] The object of the present invention is to provide a sterol
derivative having an activity to promote proliferation of neural
stem cells or a pharmaceutically acceptable salt thereof.
Means for Solving the Problems
[0012] The present invention relates to the following (1) to
(16).
(1) A sterol derivative represented by the general formula (I):
##STR00003##
[0013] [wherein Y represents optionally substituted lower alkyl or
optionally substituted lower alkenyl,
[0014] X.sup.a and X.sup.b are the same or different, and represent
a bond or --NR.sup.a-- (wherein R.sup.a represents a hydrogen atom,
optionally substituted lower alkyl, optionally substituted
cycloalkyl, optionally substituted lower alkanoyl, or optionally
substituted aroyl),
[0015] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.7 and R.sup.8 are
the same or different, and represent a hydrogen atom or hydroxy,
or
[0016] R.sup.1 and R.sup.2, R.sup.3 and R.sup.4, or R.sup.7 and
R.sup.8 together represent .dbd.O,
[0017] R.sup.5 and R.sup.6 are the same or different, and represent
a hydrogen atom, halogen, azido, hydroxy, optionally substituted
lower alkoxy, optionally substituted cycloalkyloxy, optionally
substituted lower alkenyloxy, optionally substituted lower
alkynyloxy, optionally substituted aryloxy, optionally substituted
aromatic heterocyclyloxy, optionally substituted aliphatic
heterocyclyloxy, optionally substituted lower alkanoyloxy,
optionally substituted aroyloxy, optionally substituted aromatic
heterocyclylcarbonyloxy, or --NR.sup.bR.sup.c (wherein R.sup.b and
R.sup.c are the same or different, and represent a hydrogen atom,
optionally substituted lower alkyl, optionally substituted lower
alkenyl, optionally substituted cycloalkyl, optionally substituted
aryl, an optionally substituted aromatic hetercyclic group, an
optionally substituted aliphatic heterocyclic group, optionally
substituted lower alkanoyl, optionally substituted aroyl,
optionally substituted lower alkoxycarbonyl, optionally substituted
aryloxycarbonyl, optionally substituted aromatic
heterocyclyloxycarbonyl, optionally substituted lower
alkylcarbamoyl, optionally substituted di-lower alkylcarbamoyl,
optionally substituted arylcarbamoyl, optionally substituted
aromatic heterocyclylcarbamoyl, optionally substituted
arylsulfonyl, or optionally substituted lower alkylsulfonyl), or
R.sup.5 and R.sup.6 together represent O.dbd., R.sup.dON.dbd.
(wherein R.sup.d represents a hydrogen atom, optionally substituted
lower alkyl, optionally substituted cycloalkyl, optionally
substituted aryl, an optionally substituted aromatic hetercyclic
group, or an optionally substituted aliphatic heterocyclic group),
or R.sup.eR.sup.fC.dbd. (wherein R.sup.e and R.sup.f are the same
or different, and represent a hydrogen atom, halogen, optionally
substituted lower alkyl, optionally substituted lower alkenyl,
optionally substituted lower alkynyl, optionally substituted
cycloalkyl, optionally substituted aryl, an optionally substituted
aromatic heterocyclic group, an optionally substituted aliphatic
heterocyclic group, optionally substituted lower alkoxy, optionally
substituted aryloxy, optionally substituted aromatic
heterocyclyloxy, optionally substituted lower alkanoyl, optionally
substituted aroyl, optionally substituted lower alkanoyloxy, or
optionally substituted aroyloxy),
[0018] R.sup.9 represents a hydrogen atom, optionally substituted
lower alkyl, optionally substituted lower alkenyl, optionally
substituted lower alkynyl, or optionally substituted cycloalkyl,
or
[0019] R.sup.1 and R.sup.3, R.sup.3 and R.sup.5 (provided that this
is only when X.sup.a is a bond), R.sup.5 and R.sup.7 (provided that
this is only when X.sup.b is a bond), or R.sup.7 and R.sup.9
together represent a bond or an oxygen atom, and
[0020] regarding R.sup.10, R.sup.11 and R.sup.12, R.sup.10 and
R.sup.11 together represent a bond or an oxygen atom and R.sup.12
represents an oxygen atom, or R.sup.1.degree. and R.sup.12 together
represent a bond or an oxygen atom and R.sup.H represents a
hydrogen atom] (provided that compounds represented by the
following formulas (P) and (Q) are excluded), or a pharmaceutically
acceptable salt thereof.
##STR00004##
(2) The sterol derivative or the pharmaceutically acceptable salt
thereof described in (1), wherein R.sup.10 and R.sup.11 together
represent a bond and R.sup.12 is a hydrogen atom. (3) The sterol
derivative or the pharmaceutically acceptable salt thereof
described in (1) or (2), wherein X.sup.a and X.sup.b are bonds. (4)
The sterol derivative or the pharmaceutically acceptable salt
thereof described in any one of (1) to (3), wherein R.sup.7 and
R.sup.8 are hydrogen atoms. (5) The sterol derivative or the
pharmaceutically acceptable salt thereof described in any one of
(1) to (4), wherein R.sup.5 is a hydrogen atom. (6) The sterol
derivative or the pharmaceutically acceptable salt thereof
described in any one of (1) to (5), wherein R.sup.6 is hydroxy,
optionally substituted lower alkoxy, optionally substituted
cycloalkyloxy, optionally substituted lower alkenyloxy, optionally
substituted lower alkynyloxy, optionally substituted aryloxy,
optionally substituted aromatic heterocyclyloxy, optionally
substituted aliphatic heterocyclyloxy, optionally substituted lower
alkanoyloxy, optionally substituted aroyloxy, or
--NR.sup.b1R.sup.c1 (wherein R.sup.b1 and R.sup.c1 are the same or
different, and represent a hydrogen atom, optionally substituted
lower alkyl, optionally substituted lower alkanoyl, or optionally
substituted aroyl). (7) The sterol derivative or the
pharmaceutically acceptable salt thereof described in any one of
(1) to (5), wherein R.sup.6 is hydroxy, optionally substituted
lower alkoxy, optionally substituted aryloxy, optionally
substituted aromatic heterocyclyloxy, optionally substituted
aliphatic heterocyclyloxy, or --NR.sup.b1R.sup.c1 (wherein R.sup.b1
and R.sup.c1 have the same meanings as defined above, respectively)
(8) The sterol derivative or the pharmaceutically acceptable salt
thereof described in any one of (1) to (5), wherein R.sup.6 is
hydroxy or optionally substituted lower alkoxy. (9) The sterol
derivative or the pharmaceutically acceptable salt thereof
described in any one of (1) to (4), wherein R.sup.5 and R.sup.6
together represent O.dbd. or R.sup.dON.dbd. (wherein R.sup.d has
the same meaning as defined above). (10) The sterol derivative or
the pharmaceutically acceptable salt thereof described in any one
of (1) to (9), wherein R.sup.9 is a hydrogen atom or optionally
substituted lower alkyl. (11) The sterol derivative or the
pharmaceutically acceptable salt thereof described in any one of
(1) to (10), wherein Y is optionally substituted lower alkyl. (12)
The sterol derivative or the pharmaceutically acceptable salt
thereof described in any one of (1) to (10), wherein Y is lower
alkyl. (13) A proliferation promoting agent for neutral stem cells,
comprising the compound or the pharmaceutically acceptable salt
thereof described in any one of (1) to (12) as an active
ingredient. (14) A method for promoting proliferation of neutral
stem cells, comprising administering an effective amount of the
compound or the pharmaceutically acceptable salt thereof described
in any one of (1) to (12). (15) The compound or the
pharmaceutically acceptable salt thereof described in any one of
(1) to (12) for use in promoting proliferation of neural stem
cells. (16) Use of the compound or the pharmaceutically acceptable
salt thereof described in any one of (1) to (12) for manufacture of
a proliferation promoting agent for neural stem cells.
Effects of the Invention
[0021] The present invention can provide a sterol derivative having
an activity to promote proliferation of neural stem cells or a
pharmaceutically acceptable salt thereof.
EMBODIMENTS FOR CARRYING OUT THE INVENTION
[0022] Hereinafter, a compound represented by the general formula
(I) will be referred to as compound (I). The same applies to
compounds of other formula numbers.
[0023] In the definition of each group in the general formula (I),
examples of the lower alkyl, and the lower alkyl moiety of the
lower alkoxy, the lower alkanoyl, the lower alkoxycarbonyl, the
lower alkanoyloxy, the lower alkylcarbamoyl, the di-lower
alkylcarbamoyl and the lower alkylsulfonyl may include, for
example, linear or branched alkyl having 1 to 10 carbon atoms, and
more specific examples thereof may include methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
isopentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl and the
like. Two lower alkyl moieties of the di-lower alkylcarbamoyl may
be the same or different.
[0024] Examples of the lower alkenyl, and the lower alkenyl moiety
of the lower alkenyloxy may include, for example, linear or
branched alkenyl having 2 to 10 carbon atoms, and more specific
examples thereof may include vinyl, allyl, 1-propenyl, 1-butenyl,
2-butenyl, 3-butenyl, pentenyl, hexenyl, heptenyl, octenyl,
nonenyl, decenyl and the like.
[0025] Examples of the lower alkynyl, and the lower alkynyl moiety
of the lower alkynyloxy may include, for example, linear or
branched alkynyl having 2 to 10 carbon atoms, and more specific
examples thereof may include ethynyl, propynyl, butynyl, pentynyl,
hexynyl, heptynyl, octynyl, nonynyl, decynyl and the like.
[0026] Examples of the cycloalkyl, and the cycloalkyl moiety of the
cycloalkyloxy may include, for example, cycloalkyl having 3 to 8
carbon atoms, and more specific examples thereof may include
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl and the like.
[0027] Examples of the aryl, and the aryl moiety of the aryloxy,
the aroyl, the aroyloxy, the aryloxycarbonyl, the arylcarbamoyl and
the arylsulfonyl may include, for example, aryl having 6 to 14
carbon atoms, and more specific examples thereof may include
phenyl, naphtyl, azulenyl, anthryl and the like.
[0028] Examples of the aliphatic heterocyclic group, and the
aliphatic heterocyclic group moiety of the aliphatic
heterocyclyloxy may include a 5-membered or 6-membered monocyclic
aliphatic heterocyclic group including at least one atom selected
from a nitrogen atom, an oxygen atom and a sulfur atom, a bicyclic
or tricyclic condensed aliphatic heterocyclic group containing at
least one atom selected from a nitrogen atom, an oxygen atom and a
sulfur atom, wherein 3- to 8-membered rings are condensed, and the
like, and more specific examples thereof may include aziridinyl,
azetidinyl, pyrrolidinyl, piperidino, piperidinyl, azepanyl,
1,2,5,6-tetrahydropyridyl, imidazolidinyl, pyrazolidinyl,
piperazinyl, homopiperazinyl, pyrazolinyl, oxiranyl,
tetrahydrofuranyl, tetrahydro-2H-pyranyl, 5,6-dihydro-2H-pyranyl,
oxazolidinyl, morpholino, morpholinyl, thioxazolidinyl,
thiomorpholinyl, 2H-oxazolyl, 2H-thioxazolyl, dihydroindolyl,
dihydroisoindolyl, dihydrobenzofuranyl, benzoimidazolidinyl,
dihydrobenzooxazolyl, dihydrobenzothioxazolyl, tetrahydroquinolyl,
tetrahydroisoquinolyl, dihydro-2H-chromanyl, dihydro-1H-chromanyl,
dihydro-2H-thiochromanyl, dihydro-1H-thiochromanyl,
tetrahydroquinoxalinyl, tetrahydroquinazolinyl,
dihydrobenzodioxanyl and the like.
[0029] Examples of the aromatic heterocyclic group, and the
aromatic heterocyclic group moiety of the aromatic heterocyclyloxy,
the aromatic heterocyclylcarbonyloxy, the aromatic
heterocyclyloxycarbonyl and the aromatic heterocyclylcarbamoyl may
include a 5-membered or 6-membered monocyclic aromatic heterocyclic
group containing at least one atom selected from a nitrogen atom,
an oxygen atom, and a sulfur atom, a bicyclic or tricyclic
condensed aromatic heterocyclic group containing at least one atom
selected from a nitrogen atom, an oxygen atom and a sulfur atom,
wherein 3- to 8-membered rings are condensed, and the like, and
more specific examples thereof may include furyl, thienyl,
pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,
thiazolyl, isothiazolyl, thiadiazolyl, triazolyl, tetrazolyl,
pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl,
benzofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl,
isoindolyl, indolyl, indazolyl, benzoimidazolyl, benzotriazolyl,
oxazolopyrimidinyl, thiazolopyrimidinyl, pyrrolopyridinyl,
pyrrolopyrimidinyl, imidazopyridinyl, purinyl, quinolinyl,
isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl,
quinoxalinyl, naphthyridinyl and the like.
[0030] The halogen means each atom of fluorine, chlorine, bromine
or iodine.
[0031] The substituents of the optionally substituted lower alkyl,
the optionally substituted lower alkenyl, the optionally
substituted lower alkynyl, the optionally substituted lower alkoxy,
the optionally substituted lower alkenyloxy, the optionally
substituted lower alkynyloxy, the optionally substituted lower
alkanoyloxy, the optionally substituted lower alkanoyl, the
optionally substituted lower alkoxycarbonyl, the optionally
substituted lower alkylcarbamoyl, the optionally substituted
di-lower alkylcarbamoyl and the optionally substituted lower
alkylsulfonyl), may be the same or different, and may include, for
example, 1 to 3 substituent(s) selected from the group consisting
of halogen; hydroxy; sulfanyl; nitro; azido; cyano; carboxyl;
carbamoyl; formyl; C.sub.3-8 cycloalkyl; C.sub.6-14 aryl optionally
substituted by 1 to 3 substituent(s) selected from the group
consisting of halogen, hydroxy, amino, nitro, carboxyl, C.sub.1-10
alkoxycarbonyl, C.sub.1-10 alkoxy and trifluoromethyl; an aliphatic
heterocyclic group; an aromatic heterocyclic group; C.sub.1-10
alkoxy optionally substituted by 1 to 3 substituent(s) selected
from the group consisting of halogen, hydroxy, amino, carboxyl,
C.sub.1-10 alkoxy, C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino
and C.sub.1-10 alkoxycarbonyl; C.sub.2-10 alkenyloxy; C.sub.3-8
cycloalkoxy; C.sub.6-14 aryloxy optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; C.sub.7-16 aralkyloxy
optionally substituted by 1 to 3 substituent(s) selected from the
group consisting of halogen, hydroxy, amino, nitro, carboxyl,
C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and trifluoromethyl;
C.sub.2-11 alkanoyloxy; C.sub.7-15 aroyloxy; C.sub.1-10
alkylsulfonyloxy; trifluoromethanesulfonyloxy; C.sub.6-14
arylsulfonyloxy; p-toluenesulfonyloxy; C.sub.1-10 alkylsulfanyl;
C.sub.6-14 arylsulfanyl; --NR.sup.XaR.sup.Ya (wherein R.sup.Xa and
R.sup.Ya are the same or different, and represent a hydrogen atom;
formyl; C.sub.1-10 alkyl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino,
di-C.sub.1-10 alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.3-8
cycloalkyl; C.sub.6-14 aryl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; an aromatic heterocyclic
group; C.sub.7-16 aralkyl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; C.sub.2-11 alkanoyl
optionally substituted by 1 to 3 substituent(s) selected from the
group consisting of halogen, hydroxy, amino, carboxyl, C.sub.1-10
alkoxy, C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino and
C.sub.1-10 alkoxycarbonyl; C.sub.7-15 aroyl; C.sub.1-10
alkoxycarbonyl; C.sub.7-16 aralkyloxycarbonyl; C.sub.1-10
alkylcarbamoyl, di-C.sub.1-10 alkylcarbamoyl, C.sub.6-14
arylcarbamoyl, C.sub.1-10 alkylsulfonyl; trifluoromethanesulfonyl;
C.sub.6-14 arylsulfonyl or p-toluenesulfonyl); C.sub.2-11 alkanoyl;
C.sub.3-8 cycloalkylcarbonyl; C.sub.7-15 aroyl; aliphatic
heterocyclylcarbonyl; aromatic heterocyclylcarbonyl; C.sub.1-10
alkoxycarbonyl; C.sub.6-14 aryloxycarbonyl; C.sub.7-16
aralkyloxycarbonyl; C.sub.1-10 alkylcarbamoyl optionally
substituted by 1 to 3 substituent(s) selected from the group
consisting of hydroxy, halogen, C.sub.1-10 alkoxy, amino,
C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino, C.sub.2-11
alkanoylamino and C.sub.1-10 alkoxycarbonylamino; di-C.sub.1-10
alkylcarbamoyl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of hydroxy, halogen, C.sub.1-10
alkoxy, amino, C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino,
C.sub.2-11 alkanoylamino and C.sub.1-10 alkoxycarbonylamino; and
C.sub.6-14 arylcarbamoyl.
[0032] The substituents of the optionally substituted aryl, the
optionally substituted aryloxy, the optionally substituted aroyl,
the optionally substituted aroyloxy, the optionally substituted
aryloxycarbonyl, the optionally substituted arylcarbamoyl, the
optionally substituted arylsulfonyl, the optionally substituted
aromatic heterocyclic group, the optionally substituted aromatic
heterocyclyloxy, the optionally substituted aromatic
heterocyclylcarbonyloxy, the optionally substituted aromatic
heterocyclyloxycarbonyl, and the optionally substituted aromatic
heterocyclylcarbamoyl may be the same or different, and may
include, for example, 1 to 5 substituent(s) selected from the group
consisting of halogen; hydroxy; sulfanyl; nitro; cyano; carboxyl;
carbamoyl; C.sub.1-10 alkyl; trifluoromethyl; C.sub.3-8 cycloalkyl;
C.sub.6-14 aryl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of halogen, hydroxy, amino,
nitro, carboxyl, C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and
trifluoromethyl; an aliphatic heterocyclic group; an aromatic
heterocyclic group; C.sub.1-10 alkoxy optionally substituted by 1
to 3 substituent(s) selected from the group consisting of halogen,
hydroxy, amino, carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino,
di-C.sub.1-10 alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.3-8
cycloalkoxy; C.sub.6-14 aryloxy optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; C.sub.7-16 aralkyloxy
optionally substituted by 1 to 3 substituent(s) selected from the
group consisting of halogen, hydroxy, amino, nitro, carboxyl,
C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and trifluoromethyl;
C.sub.2-11 alkanoyloxy; C.sub.7-15 aroyloxy; C.sub.1-10
alkylsulfonyloxy; trifluoromethanesulfonyloxy; C.sub.6-14
arylsulfonyloxy; p-toluenesulfonyloxy; C.sub.1-10 alkylsulfanyl;
C.sub.6-14 arylsulfanyl; --NR.sup.XbR.sup.Yb (wherein R.sup.Xb and
R.sup.Yb are the same or different, and represent a hydrogen atom;
formyl; C.sub.1-10 alkyl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino,
di-C.sub.1-10 alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.3-8
cycloalkyl; C.sub.6-14 aryl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; an aromatic heterocyclic
group; C.sub.7-16 aralkyl optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; C.sub.2-11 alkanoyl
optionally substituted by 1 to 3 substituent(s) selected from the
group consisting of halogen, hydroxy, amino, carboxyl, C.sub.1-10
alkoxy, C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino and
C.sub.1-10 alkoxycarbonyl; C.sub.7-15 aroyl; C.sub.1-10
alkoxycarbonyl; C.sub.7-16 aralkyloxycarbonyl; C.sub.1-10
alkylsulfonyl; trifluoromethanesulfonyl; C.sub.6-14 arylsulfonyl or
p-toluenesulfonyl); C.sub.2-11 alkanoyl; C.sub.3-8
cycloalkylcarbonyl; C.sub.7-15 aroyl; aliphatic
heterocyclylcarbonyl; aromatic heterocyclylcarbonyl; C.sub.1-10
alkoxycarbonyl; C.sub.6-14 aryloxycarbonyl; C.sub.7-16
aralkyloxycarbonyl; C.sub.1-10 alkylcarbamoyl; di-C.sub.1-10
alkylcarbamoyl; C.sub.6-14 arylcarbamoyl; C.sub.1-10 alkylsulfonyl;
and C.sub.6-14 arylsulfonyl.
[0033] The substituents of the optionally substituted cycloalkyl,
the optionally substituted cycloalkyloxy, the optionally
substituted aliphatic heterocyclic group and the optionally
substituted aliphatic heterocyclyloxy may be the same or different,
and may include, for example, 1 to 5 substituent(s) selected from
the group consisting of oxo; halogen; hydroxy; sulfanyl; nitro;
cyano; carboxyl; carbamoyl; C.sub.1-10 alkyl optionally substituted
by 1 to 3 hydroxy groups; trifluoromethyl; C.sub.3-8 cycloalkyl;
C.sub.6-14 aryl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of halogen, hydroxy, amino,
nitro, carboxyl, C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and
trifluoromethyl; an aliphatic heterocyclic group; an aromatic
heterocyclic group; C.sub.1-10 alkoxy optionally substituted by 1
to 3 substituent(s) selected from the group consisting of halogen,
hydroxy, amino, carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino,
di-C.sub.1-10 alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.3-8
cycloalkoxy; C.sub.6-14 aryloxy optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl;
[0034] C.sub.7-16 aralkyloxy optionally substituted by 1 to 3
substituent(s) selected from the group consisting of halogen,
hydroxy, amino, nitro, carboxyl, C.sub.1-10 alkoxycarbonyl,
C.sub.1-10 alkoxy and trifluoromethyl; C.sub.2-11 alkanoyloxy;
C.sub.7-15 aroyloxy; C.sub.1-10 alkylsulfonyloxy;
trifluoromethanesulfonyloxy; C.sub.6-14 arylsulfonyloxy;
p-toluenesulfonyloxy; C.sub.1-10 alkylsulfanyl; C.sub.6-14
arylsulfanyl; --NR.sup.XcR.sup.Yc (wherein R.sup.Xc and R.sup.Yc
are the same or different, and represent a hydrogen atom; formyl;
C.sub.1-10 alkyl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of halogen, hydroxy, amino,
carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino, di-C.sub.1-10
alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.3-8 cycloalkyl;
C.sub.6-14 aryl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of halogen, hydroxy, amino,
nitro, carboxyl, C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and
trifluoromethyl; an aromatic heterocyclic group; C.sub.7-16 aralkyl
optionally substituted by 1 to 3 substituent(s) selected from the
group consisting of halogen, hydroxy, amino, nitro, carboxyl,
C.sub.1-10 alkoxycarbonyl, C.sub.1-10 alkoxy and trifluoromethyl;
C.sub.2-11 alkanoyl optionally substituted by 1 to 3 substituent(s)
selected from the group consisting of halogen, hydroxy, amino,
carboxyl, C.sub.1-10 alkoxy, C.sub.1-10 alkylamino, di-C.sub.1-10
alkylamino and C.sub.1-10 alkoxycarbonyl; C.sub.7-15 aroyl;
alkoxycarbonyl; C.sub.7-16 aralkyloxycarbonyl; C.sub.1-10
alkylsulfonyl; trifluoromethanesulfonyl; C.sub.6-14 arylsulfonyl or
p-toluenesulfonyl), C.sub.2-11 alkanoyl; C.sub.3-8
cycloalkylcarbonyl; C.sub.7-15 aroyl; aliphatic
heterocyclylcarbonyl; aromatic heterocyclylcarbonyl; C.sub.1-10
alkoxycarbonyl; C.sub.6-14 aryloxycarbonyl; C.sub.7-16
aralkyloxycarbonyl; C.sub.1-10 alkylcarbamoyl; di-C.sub.1-10
alkylcarbamoyl; C.sub.6-14 arylcarbamoyl; C.sub.1-10 alkylsulfonyl;
and C.sub.6-14 arylsulfonyl.
[0035] Examples of the C.sub.1-10 alkyl, and the C.sub.1-10 alkyl
moiety of the C.sub.1-10 alkoxy, the C.sub.2-11 alkanoyloxy, the
C.sub.1-10 alkylsulfanyl, the di-C.sub.1-10 alkylamino, the
C.sub.2-11 alkanoylamino, the C.sub.1-10 alkoxycarbonylamino, the
C.sub.2-11 alkanoyl, the C.sub.1-10 alkoxycarbonyl, the C.sub.1-10
alkylcarbamoyl, the di-C.sub.1-10 alkylcarbamoyl, the C.sub.1-10
alkylsulfonyl, and the C.sub.1-10 alkylsulfonyloxy as mentioned
herein include the groups recited as examples of the aforementioned
lower alkyl. Two C.sub.1-10 alkyl groups of the di-C.sub.1-10
alkylamino and the di-C.sub.1-10 alkylcarbamoyl may be the same or
different.
[0036] Examples of the C.sub.2-10 alkenyl moiety of the C.sub.2-10
alkenyloxy include the groups recited as examples of the
aforementioned lower alkenyl.
[0037] Examples of the C.sub.3-8 cycloalkyl, and the C.sub.3-8
cycloalkyl moiety of the C.sub.3-8 cycloalkoxy and the C.sub.3-8
cycloalkylcarbonyl include the groups recited as examples of the
aforementioned cycloalkyl.
[0038] Examples of the C.sub.6-14 aryl, and the C.sub.6-14 aryl
moiety of the C.sub.6-14 aryloxy, the C.sub.7-15 aroyloxy, the
C.sub.6-14 arylsulfanyl, the C.sub.6-14 arylsulfonyl, the
C.sub.7-15 aroyl, the C.sub.6-14 aryloxycarbonyl, the C.sub.6-14
aryloxycarbamoyl, the C.sub.6-14 arylsulfonyl and the C.sub.6-14
arylsulfonyloxy include the groups recited as examples of the
aforementioned aryl.
[0039] Examples of the aryl moiety of the C.sub.7-16 aralkyloxy,
the C.sub.7-16 aralkyl and the C.sub.7-16 aralkyloxycarbonyl
include the groups recited as examples of the aforementioned aryl,
examples of the alkyl moiety thereof include C.sub.1-10 alkylene,
and more specifically, may include a group obtained by removing one
hydrogen atom from the groups recited as examples of the
aforementioned lower alkyl.
[0040] Examples of the aliphatic heterocyclic group and the
aliphatic heterocyclic group moiety of the aliphatic
heterocyclylcarbonyl, the aromatic heterocyclic group and the
aromatic heterocyclic group moiety of the aromatic
heterocyclylcarbonyl, and the halogen may include the groups
recited as examples of the aforementioned aliphatic heterocyclic
group, the aforementioned aromatic heterocyclic group, and the
aforementioned halogen, respectively.
[0041] In the general formula (I), in addition to the
aforementioned (1) to (12), Y is, for example, preferably
C.sub.1-10 alkyl, and more preferably C.sub.1-10 alkyl which is
substituted at position 2 (the following formula (II)), and the
like. These preferable group may optionally substituted with
hydroxy, C.sub.1-10 alkoxy, C.sub.2-10 alkenyloxy, C.sub.2-11
alkanoyloxy, C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino and
the like, and preferably, may be substituted with, for example,
hydroxy, C.sub.1-4 alkoxy, C.sub.2-4 alkenyloxy, C.sub.2-5
alkanoyloxy, C.sub.1-4 alkylamino, alkylamino, and the like.
##STR00005##
[0042] (wherein Y.sup.A represents C.sub.1-8 alkyl)
[0043] R.sup.5 is, for example, preferably a hydrogen atom and the
like, and
[0044] R.sup.6 is, for example, preferably hydroxy, C.sub.1-10
alkoxy, C.sub.6-14 aryloxy, aliphatic heterocyclyloxy, aromatic
heterocyclyloxy, amino, C.sub.1-10 alkylamino, C.sub.2-11
alkanolylamino, C.sub.6-14 arylcarbonylamino, and the like, more
preferably hydroxy, C.sub.1-10 alkoxy, and the like, and still more
preferably hydroxy, C.sub.1-4 alkoxy, and the like. Further, groups
thereof may be substituted with 1 to 3 substituent(s), and the
substituent(s) thereof include, for example, hydroxy, C.sub.1-10
alkoxy, C.sub.2-10 alkenyloxy, C.sub.2-11 alkanoyloxy, amino,
C.sub.1-10 alkylamino, di-C.sub.1-10 alkylamino, C.sub.2-11
alkanoylamino, pyrrolidinyl, piperidino, morpholino, and the like,
preferably, for example, hydroxy, C.sub.1-10 alkoxy, and the like,
still more preferably, for example, hydroxy, C.sub.1-4 alkoxy and
the like, and further more preferably, for example, hydroxy,
methoxy, and the like.
[0045] R.sup.9 is, for example, preferably a hydrogen atom,
C.sub.1-10 alkyl, C.sub.1-10 alkyl substituted with 1 to 2 hydroxy
group(s), and the like, more preferably a hydrogen atom, C.sub.1-4
alkyl, C.sub.1-4 alkyl substituted with 1 to 2 hydroxy group(s),
and the like, and still more preferably a hydrogen atom, and the
like.
[0046] Example of the pharmaceutically acceptable salt of Compound
(I) include pharmaceutically acceptable acid addition salts, metal
salts, ammonium salts, organic amine addition salts, amino acid
addition salts, and the like. Examples of the pharmaceutically
acceptable acid addition salt of Compound (I) include an inorganic
salt such as a hydrochloride, a hydrobromate, a nitrate, a sulfate
and a phosphate, an organic acid salt such as an acetate, an
oxalate, a maleate, a fumarate, a citrate, a benzoate, a
methanesulfonate and the like. Examples of the pharmaceutically
acceptable metal salt include an alkaline metal salt such as a
sodium salt and a potassium salt, an alkaline-earth metal salt such
as a magnesium salt and a calcium salt, an aluminium salt, a zinc
salt, and the like. Examples of the pharmaceutically acceptable
ammonium salt include a salt of ammonium, tetramethylammonium, and
the like. Examples of the pharmaceutically acceptable organic amine
addition salt include an addition salt of morpholine, piperidine,
or the like. Examples of the pharmaceutically acceptable amino acid
addition salt include an addition salt of lysine, glycine,
phenylalanine, asparagic acid, glutamic acid, and the like.
[0047] Next, a preparation method of the compound of the present
invention is described.
[0048] In the preparing methods as shown below, when the defined
group changes under the conditions of the method carried out, or
the method is inappropriate for carrying out, the desired compound
can be obtained by using the protection and deprotection of the
groups which are ordinarily used in the synthetic organic chemistry
[e.g., Protective Groups in Organic Synthesis, third edition, T. W.
Greene, John Wiley & Sons Inc. (1999)] and the like. In
addition, the order of the steps for introducing a substituent and
the like may be changed, if necessary.
[0049] Preparation Method 1
[0050] Among compounds (I), compounds (I-a) and (I-b) in the
following scheme may be prepared, for example, according to the
following steps.
##STR00006## ##STR00007##
[0051] (wherein R.sup.22 represents a protective group for a
hydroxy group such as a trialkyl silyl group such as trimethyl
silyl, tert-butyldimethylsilyl and the like, a tetrahydropiranyl
group, a benzyl group and the like, and Y has the same meaning as
defined above)
[0052] Step 1
[0053] With 1 to 5 equivalents of a borating agent,
7-dehydrocholesterol in which 3-hydroxy group is protected
(compound (IV)) may be reacted in an inert solvent at a temperature
between -78.degree. C. and 100.degree. C. for 5 minutes to 72
hours, followed by treatment with 1 equivalent to a large excess
amount of an oxidizing agent in the presence of a suitable base at
a temperature between -78.degree. C. and 100.degree. C. for 5
minutes to 72 hours to prepare compound (V).
[0054] Examples of the inert solvent include diethyl ether,
tetrahydrofuran (THF), dioxane, N,N-dimethylformamide (DMF),
N,N-dimethylacetamide (DMA), dimethylsulfoxide (DMSO), benzene,
toluene, xylene, dichloromethane, chloroform, water and the like,
and these may be used in combination. Examples of the borating
agent include, a borane-tetrahydrofuran complex, methylborane, a
borane-dimethylsulfide complex, 9-borabicyclo[3.3.1]nonane,
catecholborane and the like.
[0055] Examples of the base may include sodium hydroxide, potassium
hydroxide, calcium hydroxide, potassium carbonate and the like.
Examples of the oxidizing agent include hydrogen peroxide, oxygen,
and the like.
[0056] Compound (IV) may be obtained by protecting 3-hydroxy group
in 7-dehydrocholesterol which is obtained as, for example, a
commercially available product with such as a trialkyl silyl group
according to methods described in, for example, T. W. Greene,
Protective Groups in Organic Synthesis, Third Edition, John Wiley
& Sons Inc. (1999), and the like. Further, compound (IV) may be
obtained according to methods described in, for example, Organic
Letters (2003), vol. 5, p. 1837-1839 or Chemistry-A European
Journal (2001), vol. 7, p. 2663-2670, and the like.
[0057] Step 2
[0058] Compound (VI) may be prepared by deprotecting compound (V)
obtained in the Step 1 according to the method described in T. W.
Greene, Protective Groups in Organic Synthesis, Third Edition, John
Wiley & Sons Inc. (1999).
[0059] Step 3
[0060] Compound (VII) may be obtained by reacting compound (VI)
obtained in the Step 2 with 1 equivalent to a large excess amount
of a suitable oxidizing agent in the presence of, if necessary, an
additive in an inert solvent at a temperature between -20.degree.
C. and 100.degree. C. for 5 minutes to 72 hours.
[0061] Examples of the inert solvent include diethyl ether, THF,
dioxane, DMF, DMA, DMSO, dichloromethane, chloroform, carbon
tetrachloride, dichloroethane, benzene, toluene, xylene, ethyl
acetate, acetic acid, propionic acid, butyric acid, trifluoroacetic
acid, water, pyridine and the like, and these may be used in
combination. Examples of the oxidizing agent include
tetrapropylammonium perruthenate, 4-methylmorpholine-N-oxide,
manganese dioxide, chromic acid, pyridinium chlorochromate,
pyridinium dichromate, potassium permanganate, sulfur
trioxide-pyridine, oxone, silver nitrate, silver oxide (I), silver
oxide (II), sodium periodate, sodium perchlorate, hydrogen peroxide
and the like, and these may be used in combination. Examples of the
additive include acetic acid, sulfuric acid, sulfamic acid,
ruthenium oxide, 2-methyl-2-butene, DMSO and the like.
[0062] Step 4
[0063] Compound (VIII) may be prepared by treating compound (VII)
obtained in the Step 3 with catalytic amount to 10 equivalents of a
suitable oxidizing agent in the presence of, if necessary, 1 to 10
equivalents of a suitable additive in an inert solvent at a
temperature between 0.degree. C. and 100.degree. C. for 1 to 24
hours. Further, a plurality of oxidizing agents may be used in
combination, if necessary.
[0064] Examples of the inert solvent include acetone, diethyl
ether, THF, acetonitrile, ethyl acetate, acetic acid, propionic
acid, butyric acid, trifluoroacetic acid, water, toluene, benzene,
dichloromethane and the like, and these may be used in combination.
Examples of the oxidizing agent include sodium periodate, hydrogen
peroxide, peracetic acid, performic acid, pertrifluoroacetic acid,
orthosulfoperbenzoic acid, peroxyphthalic acid, monoperoxysuccinic
acid, disuccinoylperoxide, potassium permanganate, osmium
tetroxide, a silver acetate-iodine complex,
4-methylmorpholine-N-oxide, trimethylamine-N-oxide, potassium
ferricyanate and the like. Examples of the additive include
4-methylmorpholine, triethylamine, pyridine, ruthenium chloride,
cerium chloride and the like.
[0065] Step 5 Compound (IX) may be prepared by reacting compound
(VIII) obtained in the Step 4 with 1 to 10 equivalents of a
suitable oxidizing agent in the presence of 1 to 10 equivalents of
a suitable base in an inert solvent at a temperature between
-78.degree. C. and the boiling point of the solvent used for 5
minutes to 72 hours.
[0066] Examples of the oxidizing agent include lead tetraacetate,
sodium periodate, chromic acid and the like. Examples of the inert
solvent include dichloromethane, chloroform, carbon tetrachloride,
1,2-dichloroethane, toluene, ethyl acetate, acetonitrile, diethyl
ether, THF, dimetoxy ethane (DME), dioxane, DMF, DMA, N-methyl
pyrrolidone (NMP), DMSO and the like, and these may be used in
combination. Examples of the base include pyridine, lutidine,
triethylamine, diisopropylethylamine and the like.
[0067] Step 6
[0068] Compound (I-a) may be prepared by reacting compound (IX)
obtained in the Step 5 with 1 equivalent to a large excess amount
of suitable acid anhydride, acid halide or a halogenating agent in
the presence of 1 equivalent to a large excess amount of a suitable
base in an inert solvent at a temperature between -78.degree. C.
and the boiling point of the solvent used for 5 minutes to 72
hours.
[0069] Examples of the acid anhydride or acid halide include acetic
anhydride, anhydrous propionic acid, acetyl chloride, propionyl
chloride and the like, and examples of the halogenating agent
include thionyl chloride, phosphorus oxychloride, oxalyl
dichloride, methanesulfonyl chloride and the like. Examples of the
inert solvent include dichloromethane, chloroform, carbon
tetrachloride, 1,2-dichloroethane, toluene, ethyl acetate,
acetonitrile, diethyl ether, THF, DME, dioxane, DMF, DMA, NMP, DMSO
and the like, and these may be used in combination. Examples of the
base include sodium acetate, sodium carbonate, sodium bicarbonate,
potassium carbonate, sodium hydroxide, potassium hydroxide,
pyridine, lutidine, triethylamine, diisopropylethylamine and the
like.
[0070] Step 7
[0071] Compound (I-b) may be prepared by treating compound (I-a)
obtained in the Step 6 with 1 to 10 equivalents of a suitable
reducing agent in an inert solvent at a temperature between
-78.degree. C. and the boiling point of the solvent used for 5
minutes to 72 hours.
[0072] Examples of the reducing agent include lithium aluminium
hydride, diisobutyl aluminium hydride, sodium borohydride, lithium
borohydride, diisopropyl aluminium hydride, potassium
tri(sec-butyl) borohydride and the like. Examples of the inert
solvent include dichloromethane, chloroform, carbon tetrachloride,
1,2-dichloroethane, toluene, diethyl ether, THF, DME, dioxane,
acetonitrile, methanol, ethanol, propanol, DMF, DMA, NMP, DMSO and
the like, and these may be used in combination.
[0073] Preparation Method 2
[0074] Among compounds (I), compound (I-c) in the following scheme
may be prepared, for example, according to the following steps.
##STR00008##
[0075] (wherein R.sup.d and Y have the same meanings as defined
above, respectively) Step 8
[0076] Compound (I-c) may be prepared by reacting compound (I-a)
obtained in the Step 6 of Preparation Method 1 with 1 to 10
equivalents of R.sup.dONH.sub.2 in the presence of 1 equivalent to
a large excess amount of a suitable base in an inert solvent at a
temperature between 0.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0077] Examples of the inert solvent include methanol, ethanol,
propanol, dichloromethane, chloroform, carbon tetrachloride,
1,2-dichloroethane, toluene, ethyl acetate, acetonitrile, diethyl
ether, THF, DME, dioxane, DMF, DMA, NMP, DMSO and the like, and
these may be used in combination. Examples of the base include
sodium acetate, sodium carbonate, sodium bicarbonate, potassium
carbonate, sodium hydroxide, potassium hydroxide, pyridine,
lutidine, triethylamine, diisopropylethylamine and the like.
[0078] Preparation Method 3 Among compounds (I), compounds (I-d)
and (I-e) in the following scheme may be prepared, for example,
according to the following steps.
##STR00009##
[0079] (wherein Y has the same meaning as defined above)
[0080] Step 9
[0081] Compound (X) may be prepared by reacting compound (I-a)
obtained in the Step 6 of Preparation Method 1 with 1 to 10
equivalents of chlorotrimethylsilane in the presence of 1 to 10
equivalents of a suitable base in an inert solvent at a temperature
between -78.degree. C. and room temperature for 5 minutes to 24
hours.
[0082] Examples of the inert solvent include dichloromethane,
toluene, diethyl ether, THF, DME, dioxane and the like, and these
may be used in combination. Examples of the base include lithium
diisopropyl amide (LDA), lithium hexamethyl disilazide (LHMDS),
potassium hexamethyl disilazide (KHMDS), potassium tert-butoxide
and the like.
[0083] Step 10
[0084] Compound (I-d) or (I-e) may be prepared by treating compound
(X) obtained in the Step 9 in the presence of 1 to 10 equivalents
of a palladium compound in an inert solvent at a temperature
between -78.degree. C. and room temperature for 5 minutes to 24
hours.
[0085] Examples of the palladium compound include palladium (II)
acetate, bis(triphenylphosphine) palladium (II) chloride,
[1,2-bis(diphenylphosphino)ethane]palladium (II) chloride,
[1,1'-bis(diphenylphosphino)ferrocene]palladium (II) chloride and
the like. Examples of the solvent include dichloromethane,
chloroform, carbon tetrachloride, 1,2-dichloroethane, toluene,
ethyl acetate, acetonitrile, diethyl ether, THF, DME, dioxane, DMF,
DMA, NMP, DMSO and the like, and these may be used in
combination.
[0086] Step 4
[0087] Among compounds (I), compound (I-f) in the following scheme
may be prepared, for example, according to the following step.
##STR00010##
[0088] (wherein Y has the same meaning as defined above)
[0089] Process 11
[0090] Compound (I-f) may be prepared by reacting compound (I-d)
obtained in the Step 10 of Preparation Method 3 with 1 to 10
equivalents of a suitable oxidizing agent in the presence of, if
necessary, 1 equivalent to a large excess amount of a base in an
inert solvent at a temperature between -78.degree. C. and the
boiling point of the solvent used for 5 minutes to 72 hours.
[0091] Examples of the oxidizing agent include hydrogen peroxide,
tert-butylhydroperoxide (TBHP), peracetic acid, m-chloroperbenzoic
acid, perbenzoic acid, trifluoroperacetic acid and the like.
Examples of the inert solvent include dichloromethane, chloroform,
carbon tetrachloride, 1,2-dichloroethane, toluene, diethyl ether,
THF, DME, dioxane, acetonitrile, methanol, ethanol, propanol,
tert-butanol, water, DMF, DMA, NMP, DMSO and the like, and these
may be used in combination. Examples of the base include sodium
acetate, sodium carbonate, sodium bicarbonate, potassium carbonate,
sodium hydroxide, potassium hydroxide, pyridine, lutidine,
triethylamine, diisopropylethylamine and the like.
[0092] Preparation Method 5
[0093] Among compounds (I), compound (I-g) in the following scheme
may be prepared, for example, according to the following step.
##STR00011##
[0094] (wherein R.sup.e, R.sup.f, and Y have the same meanings as
defined above, respectively,
[0095] Ph represents phenyl, and D represents a chlorine atom, a
bromine atom or an iodine atom)
[0096] Step 12 Compound (I-g) may be prepared by reacting compound
(I-a) obtained in the
[0097] Step 6 of Preparation Method 1 with 1 to 10 equivalents of
compound (XI) in the presence of 1 to 10 equivalents of a base, in
an inert solvent at a temperature between -78.degree. C. and the
boiling point of the solvent used for 5 minutes to 72 hours.
[0098] Examples of the base include LDA, LHMDS, KHMDS, potassium
tert-butoxide, sodium acetate, sodium carbonate, sodium
bicarbonate, potassium carbonate, sodium hydroxide, potassium
hydroxide, pyridine, lutidine, triethylamine, diisopropylethylamine
and the like. Examples of the inert solvent include
dichloromethane, carbon tetrachloride, 1,2-dichloroethane, toluene,
diethyl ether, THF, DME, dioxane, DMF, DMA, NMP and the like, and
these may be used in combination. Compound (XI) may be obtained,
for example, as a commercially available product.
[0099] Preparation Method 6
[0100] Among compounds (I), compound (I-h) in the following scheme
may be prepared, for example, according to the following step.
##STR00012##
[0101] (wherein R.sup.b, R.sup.c and Y have the same meanings as
defined above, respectively)
[0102] Step 13
[0103] Compound (I-h) may be prepared by reacting compound (I-a)
obtained in the Step 6 of Preparation Method 1 with 1 to 10
equivalents of compound (XII) in the presence of 1 to 10
equivalents of a reducing agent in an inert solvent at a
temperature between -78.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0104] Examples of the reducing agent include sodium borohydride,
sodium triacetoxyborohydride, sodium cyanoborohydride and the like.
Examples of the inert solvent include dichloromethane, carbon
tetrachloride, 1,2-dichloroethane, toluene, diethyl ether, THF,
DME, acetonitrile, dioxane, DMF, DMA, NMP, methanol, ethanol,
propanol, acetic acid, propionic acid and the like, and these may
be used in combination. Compound (XII) may be obtained, for
example, as a commercially available product.
[0105] Preparation Method 7
[0106] Among compounds (I), compounds (I-i) and (I-j) in the
following scheme may be prepared, for example, according to the
following steps.
##STR00013##
[0107] (wherein Ph represents phenyl and Y has the same meaning as
defined above)
[0108] Step 14
[0109] Compound (I-i) may be prepared by reacting compound (I-b)
obtained in the Step 7 of Preparation Method 1 with 1 to 10
equivalents of diphenyl phosphorazidate (compound (XIII)) in the
presence of 1 to 10 equivalents of a suitable oxygen atom receptor
or a hydrogen atom receptor in an inert solvent at a temperature
between -78.degree. C. and the boiling point of the solvent used
for 5 minutes to 72 hours.
[0110] Examples of the oxygen atom receptor include
triphenylphosphine, tributylphosphine and the like, and examples of
the hydrogen atom receptor include diethyl azodicarboxylate (DEAD),
di-tert-butylazodicarboxylate, N,N,N',N'-tetramethyl
azadicarboxamide and the like. Examples of the inert solvent
include dichloromethane, carbon tetrachloride, 1,2-dichloroethane,
toluene, diethyl ether, THF, DME, acetonitrile, dioxane, DMF, DMA,
NMP and the like, and these may be used in combination. Compound
(XIII) may be obtained, for example, as a commercially available
product.
[0111] Step 15
[0112] Compound (I-j) may be prepared by treating compound (I-i)
obtained in the Step 14 with 1 to 10 equivalents of a suitable
reducing agent in an inert solvent at a temperature between
-78.degree. C. and the boiling point of the solvent used for 5
minutes to 72 hours.
[0113] Examples of the reducing agent include lithium aluminium
hydride, sodium borohydride, lithium borohydride, diisopropyl
aluminium hydride, potassium tri-(sec-butyl) borohydride,
triphenylphosphine, borane, tributyl tin hydride and the like.
Examples of the solvent include dichloromethane, chloroform, carbon
tetrachloride, 1,2-dichloroethane, toluene, diethyl ether, THF,
DME, dioxane, acetonitrile, methanol, ethanol, propanol, DMF, DMA,
NMP, DMSO, water and the like, and these may be used in
combination.
[0114] Preparation Method 8
[0115] Among compounds (I), compound (I-k) in the following scheme
may be prepared, for example, according to the following step.
##STR00014##
[0116] (wherein R.sup.28 represents a moiety of optionally
substituted lower alkyl, optionally substituted aryl, optionally
substituted lower alkoxy, optionally substituted aryloxy or
optionally substituted aromatic heterocyclyloxy in the optionally
substituted lower alkanoyl, the optionally substituted aroyl, the
optionally substituted lower alkoxycarbonyl, the optionally
substituted aryloxycarbonyl and the optionally substituted aromatic
heterocyclyloxycarbonyl in the definition of R.sup.b or R.sup.c,
respectively, and Y has the same meaning as defined above)
[0117] Step 16
[0118] Compound (I-k) may be prepared by reacting compound (I-j)
obtained in the Step 15 of Preparation Method 7 with 1 to 10
equivalents of compound (XIV) in the presence of 1 equivalent to a
large excess amount of a base in an inert solvent at a temperature
between -78.degree. C. and the boiling point of the solvent used
for 5 minutes to 72 hours.
[0119] Examples of the base include potassium tert-butoxide, sodium
acetate, sodium carbonate, sodium bicarbonate, potassium carbonate,
sodium hydroxide, potassium hydroxide, pyridine, lutidine,
triethylamine, diisopropylethylamine and the like. Examples of the
inert solvent include dichloromethane, carbon tetrachloride,
1,2-dichloroethane, toluene, diethyl ether, THF, DME, dioxane, DMF,
DMA, NMP and the like, and these may be used in combination.
Compound (XIV) may be obtained, for example, as a commercially
available product.
[0120] Preparation Method 9
[0121] Among compounds (I), compound (I-1) in the following scheme
may be prepared, for example, according to the following step.
##STR00015##
[0122] (wherein R.sup.29 represents a moiety of optionally
substituted aryl or optionally substituted lower alkyl in the
optionally substituted arylsufonyl or the optionally substituted
lower alkylsulfonyl in the definition of R.sup.b or R.sup.c,
respectively, and Y has the same meaning as defined above)
[0123] Step 17
[0124] Compound (I-1) may be prepared by reacting compound (I-j)
obtained in the Step 15 of Preparation Method 7 with 1 to 10
equivalents of compound (XV) in the presence of 1 equivalent to a
large excess amount of a base in an inert solvent at a temperature
between -78.degree. C. and the boiling point of the solvent used
for 5 minutes to 72 hours.
[0125] Examples of the base include potassium tert-butoxide, sodium
acetate, sodium carbonate, sodium bicarbonate, potassium carbonate,
sodium hydroxide, potassium hydroxide, pyridine, lutidine,
triethylamine, diisopropylethylamine and the like. Examples of the
inert solvent include dichloromethane, carbon tetrachloride,
1,2-dichloroethane, toluene, diethyl ether, THF, DME, dioxane, DMF,
DMA, NMP and the like, and these may be used in combination.
Compound (XV) may be obtained, for example, as a commercially
available product.
[0126] Preparation Method 10
[0127] Among compounds (I), compound (I-m) in the following scheme
may be prepared, for example, according to the following step.
##STR00016##
[0128] (wherein R.sup.30 represents a moiety of optionally
substituted lower alkyl, optionally substituted aryl, or an
optionally substituted aromatic heterocyclic group in the
optionally substituted lower alkylcarbamoyl, the optionally
substituted arylcarbamoyl and the optionally substituted aromatic
heterocyclylcarbamoyl in the definition of R.sup.b or R.sup.c,
respectively, and Y has the same meaning as defined above)
[0129] Process 18
[0130] Compound (I-m) may be prepared by reacting compound (I-j)
obtained in the Step 15 of Preparation Method 7 with 1 to 10
equivalents of compound (XVI) in the presence of, if necessary, 1
equivalent to a large excess amount of a base, in an inert solvent
at a temperature between -78.degree. C. and the boiling point of
the solvent used for 5 minutes to 72 hours.
[0131] Examples of the base include potassium tert-butoxide, sodium
acetate, sodium carbonate, sodium bicarbonate, potassium carbonate,
sodium hydroxide, potassium hydroxide, pyridine, lutidine,
triethylamine, diisopropylethylamine and the like. Examples of the
inert solvent include dichloromethane, carbon tetrachloride,
1,2-dichloroethane, toluene, diethyl ether, THF, DME, dioxane, DMF,
DMA, NMP and the like, and these may be used in combination.
Compound (XVI) may be obtained, for example, as a commercially
available product.
[0132] Preparation Method 11
[0133] Among compounds (I), compound (I-n) in the following scheme
may be prepared, for example, according to such as the following
step.
##STR00017##
[0134] (wherein Y has the same meaning as defined above)
[0135] Step 19
[0136] Compound (I-n) may be prepared by reacting compound (I-d)
obtained in the Step 10 of Preparation Method 3 with catalytic
amount to 10 equivalents of a suitable oxidizing agent in the
presence of, if necessary, 1 equivalent to a large excess amount of
a co-oxidizing agent in an inert solvent at a temperature between
-78.degree. C. and the boiling point of the solvent used for 5
minutes to 72 hours.
[0137] Examples of the oxidizing agent include hydrogen peroxide,
TBHP, peracetic acid, m-chloroperbenzoic acid, perbenzoic acid,
trifluoroperacetic acid, potassium permanganate, osmium tetroxide
and the like, and examples of the co-oxidizing agent include
4-methylmorpholine-N-oxide, trimethylamine-N-oxide and the like.
Examples of the inert solvent include dichloromethane, chloroform,
carbon tetrachloride, 1,2-dichloroethane, toluene, diethyl ether,
THF, DME, dioxane, acetonitrile, methanol, ethanol, propanol,
tert-butanol, water, DMF, DMA, NMP, DMSO and the like, and these
may be used in combination.
[0138] Preparation Method 12
[0139] Among compounds (I), compound (I-o) in the following scheme
may be prepared, for example, according to such as the following
step.
##STR00018##
[0140] (wherein Y has the same meaning as defined above) Step 20
Compound (I-o) may be prepared in the same manner as the Step 7
of
[0141] Preparation Method 1 by using compound (I-d) obtained in the
Step 10 of Preparation Method 3.
[0142] Preparation Method 13
[0143] Among compounds (I), compounds (I-p), (I-q) and (I-r) in the
following scheme may be prepared, for example, according to the
following step.
##STR00019##
[0144] (wherein R.sup.31 represents a moiety of optionally
substituted lower alkyl, optionally substituted cycloalkyl,
optionally substituted lower alkenyl, optionally substituted lower
alkynyl, an optionally substituted aliphatic heterocyclic group,
optionally substituted alkanoyl or optionally substituted aroyl in
the optionally substituted lower alkoxy, the optionally substituted
cycloalkyloxy, the optionally substituted lower alkenyloxy, the
optionally substituted lower alkynyloxy, the optionally substituted
aliphatic heterocyclyloxy, the optionally substituted alkanoylox,
and the optionally substituted aroyloxy in the definition of
R.sup.5 or R.sup.6, respectively, D represents a chlorine atom, a
bromine atom or an iodine atom, and Y has the same meaning as
defined above)
[0145] Step 21
[0146] Compounds (I-p), (I-q), and (I-r) may be prepared by
reacting compound (I-b) obtained in the Step 7 of Preparation
Method 1 with 1 to 10 equivalents of compound (XVII) in the
presence of 1 to 10 equivalents of a base and, if necessary, a
suitable additive in an inert solvent at a temperature between
-78.degree. C. and the boiling point of the solvent used for 5
minutes to 72 hours.
[0147] Examples of the base include sodium hydride, potassium
iodide, LDA, LHMDS, KHMDS, potassium tert-butoxide, sodium acetate,
sodium carbonate, sodium bicarbonate, potassium carbonate, sodium
hydroxide, potassium hydroxide, pyridine, lutidine, triethylamine,
diisopropylethylamine and the like. Examples of the additive
include silver oxide and the like. Examples of the inert solvent
include dichloromethane, carbon tetrachloride, 1,2-dichloroethane,
toluene, diethyl ether, THF, DME, dioxane, DMF, DMA, NMP and the
like, and these may be used in combination. Compound (XVII) may be
obtained, for example, as a commercially available product.
[0148] Preparation Method 14
[0149] Among compounds (I), compound (I-s) in the following scheme
may be prepared, for example, according to such as the following
step.
##STR00020##
[0150] (wherein Y has the same meaning as defined above)
[0151] Step 22
[0152] Compound (I-s) may be prepared by treating compound (I-f)
obtained in the Step 11 of Preparation Method 4 in the presence of
1 equivalent to a large excess amount of hydrazine and 1 equivalent
to a large excess amount of a suitable acid in an inert solvent at
a temperature between -78.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0153] Examples of the acid include acetic acid, propionic acid,
hydrochloric acid, sulfuric acid, nitric acid and the like.
Examples of the solvent include methanol, ethanol, propanol,
butanol, dichloromethane, carbon tetrachloride, 1,2-dichloroethane,
toluene, diethyl ether, THF, DME, dioxane, DMF, DMA, NMP, DMSO,
water and the like, and these may be used in combination.
[0154] Preparation Method 15
[0155] Among compounds (I), compound (I-ab) in the following scheme
may be prepared, for example, according to such as the following
step.
##STR00021##
[0156] (wherein Y has the same meaning as defined above)
[0157] Step 23
[0158] Compound (I-ab) may be prepared in the same manner as the
Step 7 of Preparation Method 1 by using compound (I-n) obtained in
the Step 19 of Preparation Method 11.
[0159] Preparation Method 16
[0160] Among compounds (I), compounds (I-ac), (I-ad), (I-acc), and
(I-add) in the following scheme may be prepared, for example,
according to such as the following steps.
##STR00022##
[0161] (wherein R.sup.a1 represents optionally substituted lower
alkyl, optionally substituted cycloalkyl, optionally substituted
lower alkanoyl, or optionally substituted aroyl in the definition
of R.sup.a, respectively, D represents a chlorine atom, a bromine
atom and an iodine atom, and Y has the same meaning as defined
above)
[0162] Step 24 Compounds (I-ac) and (I-ad) may be prepared by
treating compound (I-a) obtained in the Step 6 of Preparation
Method 1 with 1 equivalent to a large excess amount of hydroxy
amine in the presence of 1 equivalent to a large excess amount of a
suitable acid, acid chloride or acid anhydride, and, if necessary,
in the presence of a suitable base in an inert solvent at a
temperature between 0.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0163] Examples of the inert solvent include dioxane, acetonitrile,
diethyl ether, THF, DME, DMF, DMA, NMP, DMSO, dichloromethane,
chloroform, carbon tetrachloride, 1,2-dichloroethane, toluene and
the like, and these may be used in combination. Examples of the
acid include methanesulfonic acid, hydrochloric acid, phosphoric
acid, sulfuric acid, nitric acid, formic acid, acetic acid,
propionic acid and the like, examples of the acid chloride include
thionyl chloride, phosphorus oxychloride, p-toluenesulfonyl
chloride, methanesulfonyl chloride and the like, and examples of
the acid anhydride include acetic anhydride,
trifluoromethanesulfonic acid anhydride and the like. Examples of
the base include sodium acetate, sodium carbonate, sodium
bicarbonate, potassium carbonate, sodium hydroxide, potassium
hydroxide, pyridine, lutidine, triethylamine,
diisopropylethylamine, 4-dimethylaminopyridine and the like.
[0164] Step 24a
[0165] Compound (I-acc) or (I-add) may be prepared by reacting
compound (I-ac) or (I-ad) obtained in the Step 24 with 1 to 30
equivalents of compound (XA) in the presence of 1 equivalent to a
large excess amount of a suitable base in an inert solvent at a
temperature between -78.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0166] Examples of the inert solvent include dioxane, acetonitrile,
diethyl etherTHF, DME, DMF, DMA, NMP, DMSO, dichloromethane,
chloroform, carbon tetrachloride, 1,2-dichloroethane, toluene and
the like, and these may be used in combination. Examples of the
base include potassium tert-butoxide, sodium acetate, sodium
carbonate, sodium bicarbonate, potassium carbonate, sodium
hydroxide, potassium hydroxide, pyridine, lutidine, triethylamine,
diisopropylethylamine and the like.
[0167] Preparation Method 17
[0168] Among compounds (I), compound (I-ae) in the following scheme
may be prepared, for example, according to the following step.
##STR00023##
[0169] (wherein R.sup.41 represents a moiety of optionally
substituted ary or an optionally substituted aromatic heterocyclic
group in optionally substituted aryloxy and optionally substituted
aromatic heterocyclyloxy in the definition of R.sup.b or R.sup.c,
respectively, and Y has the same meaning as defined above)
[0170] Step 25
[0171] Compound (I-ae) may be prepared by reacting compound (I-b)
obtained in the Step 7 of Preparation Method 1 with 1 to 10
equivalents of compound (XVIII) in the presence of 1 to 10
equivalents of a suitable oxygen atom receptor and a hydrogen atom
receptor in an inert solvent at a temperature between -78.degree.
C. and the boiling point of the solvent used for 5 minutes to 72
hours.
[0172] Examples of the oxygen atom receptor include
triphenylphosphine, tributylphosphine and the like, and examples of
the hydrogen atom receptor include DEAD,
di-tert-butylazodicarboxylate, N,N,N',N'-tetramethyl
azadicarboxamide and the like. Examples of the inert solvent
include dichloromethane, carbon tetrachloride, 1,2-dichloroethane,
toluene, diethyl ether, THF, DME, acetonitrile, dioxane, DMF, DMA,
NMP and the like, and these may be used in combination. Compound
(XVIII) may be obtained, for example, as a commercially available
product.
[0173] Preparation Method 18
[0174] Among compounds (I), compounds (I-af) and (I-ag) in the
following scheme may be prepared, for example, according to the
following step.
##STR00024##
[0175] (wherein Y has the same meaning as defined above)
[0176] Step 26
[0177] Compounds (I-af) and (I-ag) may be prepared in the same
manner as the Step 4 of Preparation 1 or the Step 11 of Preparation
Method 4 by using compound (X) obtained in the Step 9 of
Preparation Method 3.
[0178] Preparation Method 19
[0179] Among compounds (I), compound (I-ah) in the following scheme
may be prepared, for example, according to the following step.
##STR00025##
[0180] (wherein Y has the same meaning as defined above)
[0181] Step 27
[0182] Compound (I-ah) may be prepared in the same manner as the
Step 7 of Preparation Method 1 by using compound (I-af) obtained in
the Step 26 of Preparation Method 18.
[0183] Preparation Method 20
[0184] Among compounds (I), compound (I-al) in the following scheme
may be prepared, for example, according to the following step.
##STR00026##
[0185] (wherein Y has the same meaning as defined above)
[0186] Step 28
[0187] Compound (I-al) may be prepared in the same manner as the
Step 7 of Preparation Method 1 by using compound (I-ag) obtained in
the Step 26 of Preparation Method 18.
[0188] Preparation Method 21
[0189] Among compounds (I), compound (I-am) in the following scheme
may be prepared, for example, according to the following step.
##STR00027##
[0190] (wherein R.sup.d and Y have the the same meanings as defined
above, respectively)
[0191] Step 29
[0192] Compound (I-am) may be prepared in the same manner as the
Step 8 of Preparation Method 2 by using compound (I-af) obtained in
the Step 26 of Preparation Method 18.
[0193] Preparation Method 22
[0194] Among compounds (I), compound (I-an) in the following scheme
may be prepared, for example, according to the following step.
##STR00028##
[0195] (wherein Y has the same meaning as defined above)
[0196] Step 30
[0197] Compound (I-an) may be prepared by treating compound (I-d)
obtained in the Step 10 of Preparation Method 3 with 1 equivalent
to a large excess amount of a base in the presence of 1 to 10
equivalents of a fluorinating agent in an inert solvent at a
temperature between -78.degree. C. and the boiling point of the
solvent used for 5 minutes to 72 hours.
[0198] Examples of the base include sodium hydride, potassium
iodide, LDA, LHMDS, KHMDS, potassium tert-butoxide, sodium acetate,
sodium carbonate, sodium bicarbonate, potassium carbonate, sodium
hydroxide, potassium hydroxide, pyridine, lutidine, triethylamine,
diisopropylethylamine and the like. Examples of the fluorinating
agent include fluorine gas, N-fluorobenzenesulfonimide,
N,N'-difluoro-2,2'-bipyridinium bistetrafluoroborate,
1-chloromethyl-4-fluoro-1,4-diazo[2.2.2]octanebisammonate, cesium
fluorosulfate, diethylaminosulfur trifluoride (DAST) and the like.
Examples of the inert solvent include dichloromethane, carbon
tetrachloride, 1,2-dichloroethane, toluene, diethyl ether, THF,
DME, dioxane, DMF, DMA, NMP and the like, and these may be used in
combination.
[0199] Preparation Method 23
[0200] Among compounds (I), compound (I-aq) in the following scheme
may be prepared, for example, according to the following step.
##STR00029##
[0201] (wherein Y has the same meaning as defined above)
[0202] Step 31
[0203] Compound (I-aq) may be prepared in the same manner as the
Step 11 of Preparation Method 4 by using compound (I-ap) obtained
in the Step 6 of Preparation Method 1.
[0204] Preparation Method 24
[0205] Compound (XXIV) of the following scheme may be prepared, for
example, according to the following steps.
##STR00030##
[0206] (wherein R.sup.51 represents C.sub.1-10 alkyl, and R.sup.22
and Y have the same meanings as defined above, respectively)
[0207] Step 32
[0208] Compound (XVI) may be prepared in the same manner as the
Step 3 of Preparation Method 1 by using compound (V) obtained in
the Step 1 of Preparation Method 1. Step 33
[0209] Compound (XX) may be prepared in the same manner as the Step
4 of Preparation Method 1 by using compound (XVI) obtained in the
Step 32.
[0210] Step 34
[0211] Compound (XXI) may be prepared in the same manner as the
Step 5 of Preparation Method 1 by using compound (XX) obtained in
the Step 33 in the presence of 1 equivalent to a large excess
amount of R.sup.51OH.
[0212] Step 35
[0213] Compound (XXII) may be prepared in the same manner as the
Step 2 of Preparation Method 1 by using compound (XXI) obtained in
the Step 34.
[0214] Step 36
[0215] Compound (XXIII) may be prepared in the same manner as the
Step 7 of Preparation Method 1 by using compound (XXII) obtained in
the Step 35.
[0216] Step 37 Compound (XXIV) may be prepared in the same manner
as the Step 3 of Preparation Method 1 by using compound (XXIII)
obtained in the Step 36.
[0217] The functional groups contained in R.sup.I to R.sup.12,
R.sup.a to R.sup.f, X.sup.a, X.sup.b and Z in compound (I) may be
converted by a known method (for example, the method described in
Comprehensive Organic Transformations 2nd edition, R. C. Larock,
Vch Verlagsgesellschaft Mbh (1999) and the like) or methods similar
thereto.
[0218] The intermediates and the desired compounds obtained in each
of the above-mentioned preparation methods may be isolated and
purified by applying separation purification methods usually used
in the synthetic organic chemistry, such as filtration, extraction,
washing, drying, concentration, recrystallization, various
chromatographies or the like. Further, intermediates may also be
subjected to a next reaction without particular purification.
[0219] Although some of compounds (I) may contain a geometric
isomer, a stereoisomer such as an optical isomer, a tautomer, and
the like, the present invention includes all possible isomers and
mixtures thereof including these.
[0220] When a salt of compound (I) is intended to be obtained,
compound (I) obtained in the form of a salt may be directly
purified. When it is obtained in a free form, compound (I) may be
dissolved or suspended in a suitable solvent, and an acid or base
is added thereto to form a salt, which may be isolated and
purified.
[0221] Further, while compound (I) and pharmaceutically acceptable
salts thereof may exist in the form of adducts with water or
various solvents, these adducts are also included in the present
invention.
[0222] Specific examples of compound (I) obtained by the present
invention are shown in Table 1 to Table 12. However, compound of
the present invention is not limited thereto.
TABLE-US-00001 TABLE 1 ##STR00031## Compound No. A 1 ##STR00032## 2
##STR00033## 3 ##STR00034## 4 ##STR00035## 5 ##STR00036## 6
##STR00037## 7 ##STR00038## 8 ##STR00039## 9 ##STR00040## 10
##STR00041## 11 ##STR00042## 12 ##STR00043##
TABLE-US-00002 TABLE 2 ##STR00044## Compound No. A 13 ##STR00045##
14 ##STR00046## 15 ##STR00047## 16 ##STR00048## 17 ##STR00049## 18
##STR00050## 19 ##STR00051## 20 ##STR00052## 21 ##STR00053## 22
##STR00054## 23 ##STR00055## 24 ##STR00056##
TABLE-US-00003 TABLE 3 ##STR00057## Compound No. A 25 ##STR00058##
26 ##STR00059## 27 ##STR00060## 28 ##STR00061## 29 ##STR00062## 30
##STR00063## 31 ##STR00064## 32 ##STR00065## 33 ##STR00066## 34
##STR00067## 35 ##STR00068## 36 ##STR00069##
TABLE-US-00004 TABLE 4 ##STR00070## Compound No. A 37 ##STR00071##
38 ##STR00072## 39 ##STR00073## 40 ##STR00074## 41 ##STR00075## 42
##STR00076## 43 ##STR00077## 44 ##STR00078## 45 ##STR00079## 46
##STR00080## 47 ##STR00081## 48 ##STR00082##
TABLE-US-00005 TABLE 5 ##STR00083## Compound No. A 49 ##STR00084##
50 ##STR00085## 51 ##STR00086## 52 ##STR00087## 53 ##STR00088## 54
##STR00089## 55 ##STR00090## 56 ##STR00091## 57 ##STR00092##
TABLE-US-00006 TABLE 6 ##STR00093## Compound No. Y 58 ##STR00094##
60 ##STR00095## 61 ##STR00096## 62 ##STR00097## 63 ##STR00098## 64
##STR00099## 65 ##STR00100## 66 ##STR00101## 67 ##STR00102## 95
##STR00103## 111 ##STR00104##
TABLE-US-00007 TABLE 7 ##STR00105## Compound No. Y 68 ##STR00106##
69 ##STR00107## 70 ##STR00108## 71 ##STR00109## 72 ##STR00110## 73
##STR00111## 74 ##STR00112## 75 ##STR00113## 96 ##STR00114## 98
##STR00115##
TABLE-US-00008 TABLE 8 ##STR00116## Compund No. Y 76 ##STR00117##
77 ##STR00118## 78 ##STR00119## 79 ##STR00120## 80 ##STR00121## 81
##STR00122## 82 ##STR00123## 83 ##STR00124## 97 ##STR00125##
TABLE-US-00009 TABLE 9 ##STR00126## Compound No. Y 87 ##STR00127##
88 ##STR00128## 89 ##STR00129## 90 ##STR00130##
TABLE-US-00010 TABLE 10 Compound No. 59 ##STR00131## 91
##STR00132## 92 ##STR00133## 93 ##STR00134## 94 ##STR00135##
TABLE-US-00011 TABLE 11 Compound No. 99 ##STR00136## 107
##STR00137## 110 ##STR00138## 112 ##STR00139##
TABLE-US-00012 TABLE 12 ##STR00140## Comound No. A 100 ##STR00141##
101 ##STR00142## 102 ##STR00143## 103 ##STR00144## 104 ##STR00145##
105 ##STR00146## 106 ##STR00147## 108 ##STR00148## 109
##STR00149##
[0223] When the compound of the present invention is allowed to
contact with neural stem cells in vitro, it can promote
proliferation of the neural stem cells.
[0224] The stem cell is a cell having a pluripotency which is an
ability to differentiate into a variety of cells and an ability to
self-renew of new stem cells by symmetric or asymmetric division.
On the other hand, a cell which enters a certain lineage and is
destined to carry through its differentiation after a limited
division is called progenitor cells. However, since it is difficult
to strictly distinguish neural stem cells and neural progenitor
cells or glial progenitor cells, neural stem cells referred as in
the present invention include neural progenitor cells and glial
progenitor cells.
[0225] The neural stem cell includes preferably cerebral adult
neural stem cells, but is not limited to.
[0226] The brain may include the brain of any animals, preferably
the brain of a mammal, more preferably that of rat, mouse, monkey,
human or the like.
[0227] Examples of the method for preparing adult neural stem cells
from an animal include a method in which a cerebral cell crude
extract is prepared by extracting the brain from an adult animal by
a surgical means and the adult stem cells are concentrated from the
crude extract, in accordance with the methods described, for
example, in The Journal of Neuroscience, 1999, vol. 19, p.
8487-8497, Genes and Development, 1996, vol. 10, p. 3129-3140, and
the like.
[0228] Also, examples of the method for preparing adult neural
stems cells from human include a method in which a cerebral cell
crude extract is prepared by collecting a tissue from the lateral
ventricle wall of a patient of neurological disorder by biopsy and
the adult stem cells are concentrated from the crude extract, in
accordance with the method described in Experimental Cell Research,
2003, vol. 289, p. 378-383.
[0229] The compound of the present invention can be used in a
method for producing neural stem cells comprising contacting the
compound of the present invention with the neural stem cells in
vitro, promoting the proliferation of neural stem cells by
culturing, and collecting the neural stem cells from the
culture.
[0230] When the compound of the present invention is used in vitro,
it is preferable to use the compound of the present invention or a
pharmaceutically acceptable salt thereof, by dissolving it in a
solution which can dissolve the compound or a pharmaceutically
acceptable salt thereof. Examples of the solution include water,
dimethyl sulfoxide (DMSO) and the like. Also, it can be used by
dissolving in various buffers such as phosphate buffered saline
(PBS).
[0231] When adult neural stem cells are cultured in the presence of
the compound of the present invention, it is preferable to add the
compound at a concentration of 1 pmol/L to 1 mmol/L for
approximately 6.25.times.10.sup.4 cells/cm.sup.2 of the adult
neural stem cells. Proliferation of neural stem cells can be
promoted by allowing adult neural stem cells to contact with the
compound of the present invention, followed by static culturing at
37.degree. C. for 1 to 14 days under an atmosphere of 5% CO.sub.2
while exchanging the whole volume or a partial volume of the medium
at intervals of 2 days.
[0232] The medium may be any medium as long as it is a medium which
does not obstruct proliferation promotion of neural stem cells. For
example, it is preferable to use DMEM/F12 medium (manufactured by
Invitrogen) containing 1% N-2 additives (manufactured by
Invitrogen), and the like.
[0233] Additionally, the neural stem cells prepared by the
above-mentioned culturing can be differentiated into neurons, by
carrying out static culturing at 37.degree. C. for 1 to 14 days
under an atmosphere of 5% CO.sub.2, while exchanging the whole
volume or a partial volume of a medium at intervals of 2 days, in a
medium which does not contain the compound of the present invention
but contains, for example, 1 nmol/L to 1 mmol/L of all-trans
retinoic acid, 1 nmol/L to 1 mmol/L of forskolin or 0.1 ng/mL to 1
mg/mL of platelet-derived growth factor (PDGF), or the like.
[0234] The medium may be any medium as long as it is a medium which
does not obstruct differentiation into neurons. For example, it is
preferable to use DMEM/F 12 medium (manufactured by Invitrogen)
containing 1% N-2 additives (manufactured by Invitrogen), and the
like.
[0235] Additionally, the neural stem cells prepared by the
above-mentioned culturing can be differentiated into glial cells,
by carrying out static culturing at 37.degree. C. for 1 to 14 days
under an atmosphere of 5% CO.sub.2, while exchanging the whole
volume or a partial volume of a medium at intervals of 2 days, in a
medium which does not contain the compound of the present invention
but contains, for example, 0.1 ng/mL to 1 mg/mL of leukemia
inhibitory factor (LIF), 0.1 ng/mL to 1 mg/mL of bone morphogenic
protein-2 (BMP-2), or the like.
[0236] The medium may be any medium as long as it is a medium which
does not obstruct differentiation into glial cells. For example, it
is preferable to use DMEM/F12 medium (manufactured by Invitrogen)
containing 1% N-2 additives (manufactured by Invitrogen), and the
like.
[0237] The neural stem cells, neuron or glial cells which are
prepared by the above-mentioned culturing can be used in the
treatment of a neurological disorder, by recovering them from the
medium and transplanting them into the focus of a patient of the
neurological disorder by a surgical technique. The neurological
disorder include, for example, Parkinson's disease, Alzheimer's
disease, Down syndrome, cerebrovascular disorders, stroke, spinal
cord injury, triplet repeat disease, multiple sclerosis,
amyotrophic lateral sclerosis, polyneuropathy, epilepsy, anxiety
disorder, schizophrenia, depression, manic depressive psychosis and
the like.
[0238] Next, proliferation promoting activity of typical compounds
is specifically described based on test examples. The following
test examples are provided for the exemplification purpose only.
Accordingly, the scope of the present invention is not limited to
the following test examples.
Test Example 1
Proliferation Promoting Activity on Neural Stem Cells
[0239] Rat adult neural stem cell line ANSC-7 cells prepared by the
method described in the following Reference Test Example 1 were
suspended in assay medium at a density of 1.6.times.10.sup.5
cells/mL; inoculated at 0.1 mL in a 96 well plate (manufactured by
Costar) of which surface was coated with polyornithine and laminin;
and cultured overnight at 37.degree. C. under an atmosphere of 5%
CO.sub.2. Thereafter, 50 .mu.L of the culture supernatant was
removed, and 50 .mu.L of a test compound which was serially diluted
with the assay medium to 2 times of the final concentration or DMSO
(negative control) was added to each well. For the serial dilution
of a test compound, non-specific adsorption was minimized by using
ProteoSave 96 U plates (manufactured by SUMILON), siliconized 1.5
mL tubes (manufactured by Assist) and Pitarack siliconized tips
(manufactured by Nacalai). After the culturing for 48 hours, 50
.mu.L of the culture supernatant was replaced by 50 .mu.L of the
assay medium containing 0.1% bovine fetal serum. Furthermore, after
the culturing for 48 hours, Living Cell Count Reagent SF
(manufactured by Nacalai Tesque) was added at the volume of 10
.mu.L per well to the medium. After the culturing for 3 hours at
37.degree. C. in the incubator which was set at 5% CO.sub.2, the
resulting solution was stirred for one minute and then the
absorbance at 490 nm (control wavelength 655 nm) was measured by
using Microplate Spectrophotometer Emax (manufactured by Molecular
Devices). By regarding the measured value of well to which the
cells were not inoculated as 0%, and the measured value of negative
control as 100%, relative values in the test compound addition
group were calculated.
[0240] Each of the compounds 1, 2, 3, 4, 31, 32, 33, 46, 48, 53,
54, 66, 69, 79, 81, 83, 99 and 105 showed a proliferation promoting
activity of 125% or more at 1.0 nmol/L. Also, each of the compounds
34, 36, 74, 75, 76, 80, 90 and 94 showed a proliferation promoting
activity of 125% or more at 10 nmol/L. Additionally, each of the
compounds 21, 30, 47, 72, 78 and 93 showed a proliferation
promoting activity of 125% or more at 100 nmol/L.
Reference Test Example 1
Isolation and Culturing of Adult Neural Stems Cells from Rat
Brain
[0241] After putting a 7-week-old Sprague Dawley Rat to sleep by
ether anesthesia and subsequent decapitation, the skull was cut
open from the parietal region to extract the brain. Under a
microscope, tissues including circumventricular region were
isolated from the extracted brain by using ophthalmic scissors and
tweezers. The tissues including circumventricular region were cut
into fragments of about 1 mm.sup.3 using ophthalmic scissors and
scalpels and then subjected to 30 minutes of digestion reaction at
37.degree. C. in 5 mL of Hanks' buffer (HBSS buffer, manufactured
by Invitrogen) containing 2.5 U/mL of papain, 250 U/mL of DNase
(all manufactured by Worthington, Freehold, N.J.) and 1 U/mL of a
neutral protease (Dispase, manufactured by Boehringer-Mannheim
Corp.). The mixture of cells and tissues obtained by the reaction
was washed three times with DMEM (manufactured by Invitrogen)
containing 10% fetal bovine serum (manufactured by Hyclone) and
then dissolved in the DMEM containing 10% fetal bovine serum,
followed by removing the undigested materials using a nylon mesh of
10 .mu.m.
[0242] The resulting crude cells extract was cultured overnight on
a culture dish of 10 cm, in an incubator of 37.degree. C. using
DMEM/F12 medium (manufactured by Invitrogen) containing 10% fetal
bovine serum. On the next day, the medium was replaced with
DMEM/F12 containing 1% of N-2 additives (manufactured by
Invitrogen) and 20 ng/mL of FGF 2 (manufactured by Pepro Tech) and
the culturing was started. Once in 3 days, half of the medium was
replaced with new DMEM/F 12 containing 1% of N-2 additives and 20
ng/mL of FGF 2 and the culturing was continued.
[0243] When a small colony consisting of small cells was formed, it
was treated with 1% trypsin for 30 seconds to 1 minute, and the
cells detached were harvested. The harvested cells were inoculated
on a multiple well culture dish (manufactured by Fisher Scientific)
which had been coated at room temperature overnight using 10
.mu.g/mL of polyornithine (manufactured by Sigma) and at 37.degree.
C. overnight using 5 .mu.g/mL of mouse EHS tumor-derived laminin
(Becton Dickinson), and the culturing was continued.
[0244] By continuing the above-mentioned culturing, small cells
having small protrusion and thickness were concentrated. The cells
were used as adult neural stem cells in the above-mentioned test
(Test Example 1).
[0245] Although the present invention is explained in more detail
based on examples and reference examples, the scope of the present
invention is not limited to these examples.
[0246] The proton nuclear magnetic resonance spectra (.sup.1H NMR)
used in Examples and Reference Examples were measured at 270 MHz or
300 MHz, and exchangeable protons may not be clearly observed
depending on the compound and the measurement condition. Further,
common notation is used to represent signal multiplicity. Mass
spectrometry was performed by using electrospray ionization (ESI)
or atmospheric pressure chemical ionization (APCI). Further,
ChemBioDraw Ultra Version 11 (CambridgeSoft) was used for the
nomenclature of compounds P5, P51 to P55, P59 to P70, P73 to P81,
P83 to P85, 34, 65 to 75, 81 to 83, 93, 95 to 99, 107, 110 and
111.
Reference Example 1
(S)-3-Triethylsiloxy-5,7-cholestadiene (Compound P1)
[0247] Commercially available 7-dehydrocholesterol (10.6 g, 27.6
mmol) was dissolved in dichloromethane (90 mL), and then imidazole
(3.75 g, 55.1 mmol) and chlorotriethylsilane (6.94 mL, 41.3 mmol)
were added at room temperature, followed by stirring for 12 hours.
A saturated aqueous ammonium chloride solution was added to the
reaction mixture, followed by extraction with chloroform twice. The
organic layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by silica gel column
chromatography to obtain the title compound (12.0 g, 87%).
[0248] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.61 (q, J=7.9 Hz,
6H), 0.62 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.89 (d, J=7.0 Hz, 3H),
0.95-1.02 (m, 15H), 1.05-2.15 (m, 23H), 2.32-2.38 (m, 2H),
3.52-3.66 (m, 1H), 5.39 (m, 1H), 5.56 (d, J=5.6 Hz, 1H).
Reference Example 2
(3S,5S,6S)-3-Triethylsiloxy-7-cholesten-6-ol (Compound P2)
[0249] Compound P1 (10.1 g, 20.2 mmol) obtained in Reference
Example 1 was dissolved in THF (140 mL), and then 1.0 mol/L of a
borane-THF complex/THF solution (20.2 mL, 20.2 mmol) was added at
0.degree. C., followed by stirring at room temperature for 40
minutes. The reaction mixture was cooled to 0.degree. C., and then
water (5 mL) was slowly added thereto. Subsequently, 30% aqueous
hydrogen peroxide (6.26 mL, 60.7 mmol) and 10% aqueous sodium
hydroxide solution (22.1 mL, 60.7 mmol) were added, followed by
stirring at room temperature for 1 hour. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain the
title compound (8.43 g, 80%).
[0250] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.54 (s, 3H), 0.60
(q, J=7.9 Hz, 6H), 0.83 (s, 3H), 0.85 (d, J=6.6 Hz, 3H), 0.87 (d,
J=6.6 Hz, 3H), 0.92 (d, J=6.3 Hz, 3H), 0.96 (t, J=7.9 Hz, 9H),
0.99-2.18 (m, 26H), 3.53 (m, 1H), 3.79 (m, 1H), 5.18 (m, 1H).
Reference Example 3
(3S,5S, 6S)-7-Cholesten-3,6-diol (Compound P3)
[0251] Compound P2 (9.60 g, 18.6 mmol) obtained in Reference
Example 2 was dissolved in THF (60 mL), and then 1.0 mol/L of a
tetrabutylammonium fluoride/THF solution (27.9 mL, 27.9 mmol) was
added at room temperature, followed by stirring at 50.degree. C.
for 30 minutes. After cooling to room temperature, a saturated
aqueous ammonium chloride solution was added to the reaction
mixture, followed by extraction with ethyl acetate twice. The
organic layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by silica gel column
chromatography to obtain the title compound (7.40 g, 99%).
[0252] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.54 (s, 3H), 0.84
(s, 3H), 0.85 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.92 (d,
J=6.3 Hz, 3H), 0.98-1.95 (m, 24H), 2.04 (m, 1H), 2.25 (m, 1H), 3.59
(m, 1H), 3.80 (m, 1H), 5.18 (m, 1H).
[0253] Reference Example 4: (S)-7-Cholesten-3,6-dione (Compound P4)
Compound P3 (7.40 g, 18.4 mmol) obtained in Reference Example 3 and
4-methylmorpholine N-oxide (7.54 g, 64.3 mmol) were dissolved in
dichloromethane (180 mL), and then powdered Molecular Sieves 4
.ANG. (15.0 g) and tetrapropylammonium perruthenate (194 mg, 0.551
mmol) were added at room temperature, followed by stirring for 8
hours. The reaction mixture was filtered through a celite, and then
the filtrate was concentrated under reduced pressure to yield a
residue. The residue was purified by silica gel column
chromatography to obtain the title compound (6.53 g, 89%).
[0254] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.64 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.95 (d, J=6.3 Hz, 3H),
1.00-1.24 (m, 7H), 1.26-2.70 (m, 22H), 5.78 (m, 1H)
Reference Example 5
(1S,2R)-2-Methyl-2-[(1R,3aR,5R,7aR)-7a-methyl-1-[(R)-6-methylheptan-2-3H]--
4-oxooctahydro-1H-inden-5-yl]-5-oxocyclohexanecarboxylic acid
(Compound P5)
[0255] Sodium periodate (11.4 g, 53.4 mmol) and cerium chloride
heptahydrate (3.32 g, 8.91 mmol) were dissolved in water (70 mL),
then ethyl acetate (210 mL), acetonitrile (210 mL), and compound P4
(3.55 g, 8.91 mmol) obtained in Reference Example 4 were added
thereto. The mixture was cooled to 0.degree. C., and ruthenium
chloride hydrate (554 mg, 2.67 mmol) was added thereto, followed by
stirring for 40 minutes. A saturated aqueous sodium thiosulfate
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain a crude
purified product of (5S,7S,8S)-7,8-dihydroxy-3,6-chlestadione (2.95
g). This was dissolved in pyridine (50 mL), and then lead
tetraacetate (11.3 g, 20.5 mmol) was added at 0.degree. C. thereto,
followed by stirring for 40 minutes. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain the
title compound (1.39 g, 37%).
[0256] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.62 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.94 (d, J=5.6 Hz, 3H),
0.99-1.96 (m, 19H), 2.10-2.55 (m, 7H), 2.60-2.75 (m, 2H), 3.26 (m,
1H).
Reference Example 6
3-(Methoxymethyloxy)-cholest-5,7-diene (Compound P6)
[0257] In dichloromethane (45 mL), 7-dehydrocholesterol (2.0 g, 5.2
mmol) was dissolved, then N,N-diisopropylethylamine (2.70 mL, 15.6
mmol) and chloromethylmethylether (1.00 mL, 13.2 mmol) were
sequentially added under ice-cooling, followed by stirring at room
temperature for 20 hours. The reaction mixture was poured into a
saturated aqueous ammonium chloride solution, followed by
extraction with ethyl acetate (100 mL.times.2). The organic layer
was washed with dilute aqueous hydrochloric acid and saturated
brine, dried over anhydrous sodium sulfate, and concentrated to
yield a residue. To the residue, methanol was added to triturate to
obtain the title compound (1.9 g, 83%).
[0258] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.56 (1H, m), 5.39
(1H, m), 4.71 (2H, s), 3.53 (1H, m), 3.38 (3H, s), 2.52 (1H, m),
2.32 (1H, m), 2.09 (1H, m), 1.96-1.87 (5H, m), 1.74-0.99 (17H, m),
0.94 (3H, s), 0.94 (3H, d, J=6.4 Hz), 0.87 (3H, d, J=6.6 Hz), 0.86
(3H, d, J=6.6 Hz), 0.61 (3H, s).
Reference Example 7
3-(Methoxymethyloxy)-cholest-7-en-6-ol (Compound P7)
[0259] Compound P6 (887.1 mg, 2.100 mmol) obtained in Reference
Example 6 was dissolved in THF (15 mL), and then dimethyl
sulfide-borane (1.0 mL, 10 mmol) was added dropwise under
ice-cooling thereto, followed by stirring at room temperature for 2
hours. The reaction mixture was again ice-cooled, and then water
(10 mL), 1.0 mol/L of an aqueous solution of sodium hydroxide (10
mL) and 34.5% aqueous hydrogen peroxide (3 mL) were sequentially
added dropwise, followed by stirring at room temperature for 1
hour. A saturated aqueous sodium sulfite solution was added to the
reaction mixture, followed by extraction with chloroform (75
mL.times.2). The organic layer was washed with saturated brine,
dried over anhydrous sodium sulfate, and concentrated to yield a
residue (1.1 g), which was purified by silica gel column
chromatography (10 to 40% ethyl acetate/n-hexane) to obtain the
title compound (650 mg, 70%).
[0260] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.17 (1H, br s), 4.69
(2H, m), 3.78 (1H, br d, J=8.1 Hz), 3.48 (1H, m), 3.36 (3H, s),
2.30 (1H, m), 2.03 (1H, m), 1.83-0.97 (25H, m), 0.90 (3H, d, J=6.2
Hz), 0.85 (3H, d, J=6.6 Hz), 0.85 (3H, d, J=6.6 Hz), 0.83 (3H, s),
0.53 (3H, s).
Reference Example 8
3-(Methoxymethyloxy)-cholest-7-en-6-one (Compound P8)
[0261] Compound P7 (650 mg, 1.46 mmol) obtained in Reference
Example 7 was dissolved in dichloromethane (30 mL), and then
manganese (IV) oxide (8.5 g) was added thereto, followed by
stirring at room temperature for 12 hours. The reaction mixture was
filtered by using celite, and the filtrate was concentrated,
followed by purification by silica gel column chromatography (10 to
20% ethyl acetate/n-hexane) to obtain the title compound (540 mg,
83%).
[0262] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, br s),
4.74-4.66 (2H, m), 3.53 (1H, m), 3.38 (3H, s), 2.30-0.98 (26H, m),
0.94 (3H, d, J=6.1 Hz), 0.87 (6H, d, J=6.6 Hz), 0.87 (3H, s), 0.60
(3H, s).
Reference Example 9
3-(Methoxymethyloxy)-6,8-seco-7-norcholestan-6-oic acid-8-one
(Compound P9)
[0263] Compound P8 (190.9 mg, 0.4300 mmol) obtained in Reference
Example 8 and sodium periodate (410 mg, 1.92 mmol) were suspended
in a mixed solvent of chloroform (5 mL)-acetonitrile (2.5 mL)-water
(5 mL), and then a small amount of ruthenium chloride hydrate was
added thereto, followed by stirring at room temperature for 18
hours. A small amount of isopropanol and water were added to the
reaction mixture and then stirred, followed by extraction with
chloroform (40 mL.times.3). The organic layer was sequentially
washed with a saturated aqueous sodium sulfite solution and
saturated brine, dried over anhydrous sodium sulfate, and
concentrated. The resulting residue was dissolved in toluene (10
mL), and then lead acetate (1.20 g, 2.71 mmol) was added, followed
by stirring at room temperature for 22 hours. A small amount of
isopropanol was added to the reaction mixture and stirred, followed
by filtration using celite. Water was added to the filtrate,
followed by extraction with chloroform (20 mL.times.3). The organic
layer was washed with saturated brine, dried over anhydrous sodium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (10 to 40% ethyl
acetate/n-hexane) to obtain the title compound (48.5 mg, 24%). At
that time, 68.2 mg of compound P8 (yield 36%) was recovered.
[0264] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s), 3.45
(1H, m), 3.36 (3H, s), 2.67 (1H, dd, J=12.8, 3.9 Hz), 2.55-2.44
(2H, m), 2.27-1.12 (24H, m), 1.25 (3H, s), 0.93 (3H, d, J=5.5 Hz),
0.87 (3H, d, J=6.6 Hz), 0.86 (3H, d, J=6.6 Hz), 0.59 (3H, s).
Reference Example 10
3-(Methoxymethyloxy)-ergosta-5,7,22-triene (Compound P10)
[0265] Ergosterol (35.7 g, 90.0 mmol) was dissolved in
dimethoxymethane (350 mL, 6.3 mol), and diphosphorous pentoxide
(17.9 g, 126 mmol) was added thereto, followed by stirring at room
temperature for 1 hour. Diatomaceous earth was added to the
reaction mixture, follwed by stirring for 30 minutes and
filtration, and the filtrate was concentrated. Ethyl acetate was
added to the resulting residue for trituration to obtain the title
compound (35.1 g, 89%).
[0266] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.56 (1H, m), 5.38
(1H, m), 5.12-5.27 (2H, m), 4.71 (2H, s), 3.53 (1H, m), 3.38 (3H,
s), 2.52 (1H, m), 2.32 (1H, m), 2.07-1.26 (18H, m), 1.03 (3H, d,
J=6.6 Hz), 0.94 (3H, s), 0.91 (3H, d, J=6.8 Hz), 0.84 (3H, d, J=6.8
Hz), 0.82 (3H, d, J=6.8 Hz), 0.63 (3H, s).
Reference Example 11
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-ergosta-6,22-diene (Compound P11)
[0267] Compound P10 (11.1 g, 25.3 mmol) obtained in Reference
Example 10 and phthalhydrazide (10.3 g, 63.3 mmol) were dissolved
in dichloromethane (150 mL), and a solution of lead acetate (14.6
g, 32.9 mmol) in dichloromethane (50 mL)-acetic acid (2.55 mL) was
added dropwise at 0.degree. C. thereto. After the mixture was
stirred at 0.degree. C. for 45 minutes, alumina (50 g) was added to
stop the reaction. After filtration, the filtrate was washed with
water, a saturated aqueous ammonium chloride solution and saturated
brine, sequentially, and dried over anhydrous sodium sulfate,
followed by purification by silica gel column chromatography (10 to
40% ethyl acetate/n-hexane) to obtain the title compound (8.0 g,
53%).
[0268] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.13 (2H, m), 7.68
(2H, m), 6.65 (1H, d, J=8.1 Hz), 6.27 (1H, d, J=8.1 Hz), 5.10-5.26
(2H, m), 4.72 (1H, d, J=6.6 Hz), 4.63 (1H, d, J=6.6 Hz), 4.07 (1H,
dd, J=13.8, 8.9 Hz), 3.94 (1H, dd, J=11.9, 7.7 Hz), 3.54 (1H, m),
3.34 (3H, s), 2.11-1.34 (18H, m), 1.02 (3H, s), 1.01 (3H, d, J=6.1
Hz), 0.89 (3H, d, J=6.8 Hz), 0.84 (3H, s), 0.82 (3H, d, J=7.0 Hz),
0.81 (3H, d, J=6.8 Hz).
Reference Example 12
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-pregna-6-en-22-al (Compound P11)
[0269] Compound P11 (8.00 g, 13.4 mmol) obtained in Reference
Example 11 was dissolved in a mixed solvent of dichloromethane (120
mL) and pyridine (22.0 mL, 268 mmol), followed by passing through
an ozone-oxygen stream at -78.degree. C. for 60 minutes. Residual
ozone was removed by passing through a nitrogen stream, and
dimethylsulfide (5.0 mL, 68 mmol) was added dropwise at -78.degree.
C. thereto, followed by stirring for 30 minutes while increasing
the temperature to room temperature. The reaction mixture was
washed with dilute aqueous hydrochloric acid and saturated brine,
and dried over anhydrous sodium sulfate. The residue obtained by
concentration was purified by silica gel column chromatography (20
to 80% ethyl acetate/n-hexane) to obtain the title compound (5.3 g,
74%).
[0270] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 9.56 (1H, d, J=3.5
Hz), 8.13 (2H, m), 7.70 (2H, m), 6.68 (1H, d, J=8.3 Hz), 6.25 (1H,
d, J=8.3 Hz), 4.72 (1H, d, J=6.6 Hz), 4.64 (1H, d, J=6.6 Hz),
4.10-4.00 (2H, m), 3.55 (1H, m), 3.35 (3H, s), 2.35 (1H, m),
2.31-1.98 (4H, m), 1.94-1.25 (11H, m), 1.14 (3H, d, J=6.8 Hz), 1.03
(3H, s), 0.88 (3H, s).
[0271] Further, compound P12 may also be synthesized in accordance
with methods as described in Chemistry-A European Journal, 2001,
vol. 7, p. 2663-2670.
Reference Example 13
20-Hydroxymethyl-3-(methoxymethyloxy)-pregna-5,7-diene (Compound
P13)
[0272] Compound P12 (230 mg, 0.432 mmol) obtained in Reference
Example 12 and lithium aluminium hydride (163 mg, 4.30 mmol) were
suspended in THF (50 mL), followed by stirring at 80.degree. C. for
1 hour. The mixture was cooled to 0.degree. C., and 1.0 mol/L
potassium sodium tartrate was added dropwise thereto, followed by
extraction with ethyl acetate (50 mL.times.2). The organic layer
was washed with saturated brine, dried over anhydrous sodium
sulfate, and concentrated to yield a residue (260 mg), which was
purified by silica gel column chromatography (10 to 30% ethyl
acetate/n-hexane) to obtain the title compound (116.5 mg, 72%).
[0273] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.55 (1H, m), 5.37
(1H, m), 4.70 (2H, s), 3.63 (1H, dd, J=10.5, 3.1 Hz), 3.52 (1H, m),
3.37 (1H, m), 3.37 (3H, s), 2.51 (1H, m), 2.31 (1H, m), 2.08-1.26
(17H, m), 1.06 (3H, d, J=6.6 Hz), 0.93 (3H, s), 0.63 (3H, s).
[0274] Further, compound P13 may also be synthesized in accordance
with methods as described in Organic Letters (2003), vol. 5, p.
1837-1839.
Reference Example 14
20-(Acetyloxymethyl)-3-(methoxymethyloxy)-pregna-5,7-diene
(Compound P14)
[0275] Compound P13 (116.5 mg, 0.3110 mmol) obtained in Reference
Example 13 was dissolved in dichloromethane (7 mL), and then
pyridine (1.5 mL) and acetic anhydride (1.5 mL) were added thereto,
followed by stirring at room temperature for 14 hours. The reaction
mixture was concentrated under reduced pressure, followed by
purification by silica gel column chromatography (0 to 20% ethyl
acetate/n-hexane) to obtain the title compound (122.1 mg, 94%).
[0276] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, m), 5.38
(1H, m), 4.70 (2H, s), 4.09 (1H, dd, J=3.5, 10.8 Hz), 3.79 (1H, dd,
J=10.8, 7.5 Hz), 3.53 (1H, m), 3.38 (3H, s), 2.52 (1H, m), 2.32
(1H, m), 2.27-1.24 (16H, m), 2.06 (3H, s), 1.04 (3H, d, J=6.6 Hz),
0.94 (3H, s), 0.64 (3H, s).
Reference Example 15
20-(Acetyloxymethyl)-3-(methoxymethyloxy)-pregna-7-en-6-ol
(Compound P15)
[0277] Compound P14 (122.1 mg, 0.2940 mmol) obtained in Reference
Example 14 was treated with dimethyl sulfide-borane (0.20 mL, 2.11
mmol), 1.0 mol/L of a sodium hydroxide aqueous solution (0.25 mL)
and 34.5% aqueous hydrogen peroxide (0.70 mL) in the same manner as
Reference Example 2 to obtain the title compound (85.6 mg,
67%).
[0278] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.16 (1H, br s), 4.68
(2H, m), 4.08 (1H, dd, J=10.6, 3.3 Hz), 3.76 (2H, m), 3.47 (1H, m),
3.35 (3H, s), 2.29 (1H, m), 2.03 (3H, s), 2.07-0.85 (19H, m), 1.00
(3H, d, J=6.6 Hz), 0.82 (3H, s), 0.54 (3H, s).
Reference Example 16
20-(Acetyloxymethyl)-3-(methoxymethyloxy)-pregna-7-en-6-one
(Compound P16)
[0279] Compound P15 (85.6 mg, 0.197 mmol) obtained in Reference
Example 15 was treated with manganese (IV) oxide (850 mg) in the
same manner as Reference Example 8 to obtain the title compound
(78.4 mg, 92%).
[0280] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.69 (1H, br s), 4.70
(1H, d, J=6.8 Hz), 4.64 (1H, d, J=6.8 Hz), 4.05 (1H, dd, J=10.8,
3.5 Hz), 3.77 (1H, dd, J=10.8, 7.3 Hz), 3.49 (1H, m), 3.30 (3H, s),
2.26-1.22 (19H, m), 2.02 (3H, s), 1.01 (3H, d, J=6.6 Hz), 0.84 (3H,
s), 0.59 (3H, s).
Reference Example 17
20-(Acetyloxymethyl)-3-(methoxymethyloxy)-6,8-seco-7-norpregnan-6-oic
acid-8-one (Compound P17)
[0281] Compound P16 (130 mg, 0.300 mmol) obtained in Reference
Example 16 was treated with a small amount of ruthenium chloride
hydrate, sodium periodate (210 mg, 1.08 mmol) and lead acetate (400
mg, 0.902 mmol) in the same manner as Reference Example 5 to obtain
the title compound (45.3 mg, 33%).
[0282] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.65 (2H, s), 4.04
(1H, m), 3.78 (1H, m), 3.50 (1H, m), 3.34 (3H, s), 2.50-1.99 (20H,
m), 2.04 (3H, s), 1.00 (3H, d, J=6.5 Hz), 0.93 (3H, s), 0.60 (3H,
s).
Reference Example 18
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-cholest-6-en-22-ol (Compound P18)
[0283] A THF solution (10 mL) of compound P12 (1.37 g, 2.58 mmol)
obtained in Reference Example 12 was added dropwise at -78.degree.
C. to a THF-diethyl ether solution (10 mL) of
3-methylbutylmagnesium bromide prepared from 1-bromo-3-methylbutane
(2.00 mL, 15.7 mmol) and magnesium foil (400 mg, 16.5 mmol),
followed by stirring at room temperature for 1 hour. A saturated
aqueous ammonium chloride solution was added to the reaction
mixture, followed by extraction with ethyl acetate. The organic
layer was washed with saturated brine, dried over anhydrous sodium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (10 to 70% ethyl
acetate/n-hexane) to obtain the title compound (500 mg, 32%).
[0284] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.16-8.11 (2H, m),
7.73-7.66 (2H, m), 6.66 (1H, d, J=8.1 Hz), 6.27 (1H, d, J=8.1 Hz),
4.73-4.62 (2H, m), 4.10-3.93 (2H, m), 3.64 (1H, m), 3.55 (1H, m),
3.35 (3H, s), 2.07-1.98 (4H, m), 1.97-1.25 (18H, m), 1.03 (3H, s),
0.91 (3H, d, J=6.6 Hz), 0.89 (6H, d, J=6.6 Hz), 0.85 (3H, s).
Reference Example 19
3-(Methoxymethyloxy)-cholest-5,7-dien-22-ol (Compound P19)
[0285] Compound P18 (500 mg, 0.828 mmol) obtained in Reference
Example 18 was treated with lithium aluminium hydride (350 mg, 9.22
mmol) in the same manner as Reference Example 13 to obtain the
title compound (216 mg, 59%).
[0286] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, dd, J=5.7,
2.4 Hz), 5.39 (1H, m), 4.71 (2H, s), 3.64 (1H, m), 3.53 (1H, m),
3.38 (3H, s), 2.52 (1H, m), 2.32 (1H, m), 2.10-1.08 (22H, m), 0.95
(3H, s), 0.93 (3H, d, J=6.8 Hz), 0.90 (6H, d, J=6.6 Hz), 0.63 (3H,
s).
Reference Example 20
3-(Methoxymethyloxy)-cholest-7-en-6,22-diol (Compound P20)
[0287] Compound P19 (216.4 mg, 0.4870 mmol) obtained in Reference
Example 19 was treated with 1.0 mol/L of a borane-THF complex (2.0
mL, 2.0 mmol), 34.5% aqueous hydrogen peroxide (1.0 mL, 10 mmol)
and 6 mol/L of an aqueous solution of sodium hydroxide (1 mL, 6
mmol) in the same manner as Reference Example 2 to obtain the title
compound (111.2 mg, 49%).
[0288] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.19 (1H, br s),
4.73-4.68 (2H, m), 3.80 (1H, m), 3.63 (1H, m), 3.50 (1H, m), 3.38
(3H, s), 2.31 (1H, m), 2.04-1.07 (25H, m), 0.91 (3H, d, J=6.6 Hz),
0.89 (6H, d, J=6.4 Hz), 0.85 (3H, s), 0.56 (3H, s).
Reference Example 21
3-(Methoxymethyloxy)-cholest-7-en-6-on-22-ol (Compound P21)
[0289] Compound P20 (55.0 mg, 0.119 mmol) obtained in Reference
Example 20 was treated with manganese oxide (500 mg, 5.75 mmol) in
the same manner as Reference Example 8 to obtain the title compound
(35.6 mg, 65%).
[0290] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.71 (1H, br s),
4.73-4.64 (2H, m), 3.62 (1H, m), 3.51 (1H, m), 3.36 (3H, s),
2.28-1.03 (25H, m), 0.91 (3H, d, J=6.8 Hz), 0.88 (6H, d, J=6.6 Hz),
0.85 (3H, s), 0.60 (3H, s).
Reference Example 22
3-(Methoxymethyloxy)-22-(tert-butyldimethylsilyloxy)-cholest-7-en-6-one
(Compound P22)
[0291] Compound P21 (35.6 mg, 0.0774 mmol) obtained in Reference
Example 21 was dissolved in dichloromethane (2 mL), and
2,6-lutidine (0.050 mL, 0.43 mmol) and tert-butyldimethylsilyl
trifluoromethanesulfonate (0.050 mL, 0.22 mmol) were added at
0.degree. C. thereto, followed by stirring for 1 hour. Saturated
brine was added to the reaction mixture, followed by extraction
with chloroform. The organic layer was dried over anhydrous sodium
sulfate, followed by purification by silica gel column
chromatography (5 to 20% ethyl acetate/n-hexane) to obtain the
title compound (43.5 mg, 98%).
[0292] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, br s),
4.74-4.65 (2H, m), 3.58 (1H, dd, J=7.9, 5.9 Hz), 3.52 (1H, m), 3.37
(3H, s), 2.29-0.99 (24H, m), 0.88 (3H, d, J=6.6 Hz), 0.87 (12H, s),
0.87 (6H, d, J=6.6 Hz), 0.60 (3H, s), 0.03 (3H, s), 0.02 (3H,
s).
Reference Example 23
3-(Methoxymethyloxy)-22-(tert-butyldimethylsilyloxy)-6,8-seco-7-norcholest-
-8-on-6-oic acid (Compound P23)
[0293] Compound P22 (43.5 mg, 0.0758 mmol) obtained in Reference
Example 22, sodium periodate (40.0 mg, 0.187 mmol) and cerium
chloride heptahydrate (10.0 mg, 0.00268 mmol) were suspended in a
mixed solvent of ethyl acetate (3 mL)-acetonitrile (3 mL)-water (5
mL), and then a small amount of ruthenium chloride hydrate was
added thereto, followed by stirring at room temperature for 2
hours. Anhydrous sodium sulfate was added to the mixture, followed
by filtration, and then, the filtrate was sequentially washed with
a saturated aqueous sodium sulfite solution and saturated brine,
and dried over anhydrous sodium sulfate. The residue obtained by
concentration was dissolved in toluene (1.5 mL), and potassium
carbonate (29.1 mg, 0.211 mmol) and lead acetate (54.5 mg, 0.123
mmol) were added thereto, followed by stirring at room temperature
for 30 minutes. The reaction mixture was diluted with ethyl
acetate, isopropanol (0.2 mL) and anhydrous sodium sulfate were
added thereto, and then the mixture was filtrated. The filtrate was
sequentially washed with a saturated aqueous sodium sulfite
solution and saturated brine, and dried over anhydrous sodium
sulfate. The residue obtained by concentration was purified by
silica gel column chromatography (10 to 30% ethyl acetate/n-hexane)
to obtain the title compound (9.2 mg, 21%).
[0294] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s), 3.57
(1H, dd, J=7.9, 5.9 Hz), 3.45 (1H, m), 3.36 (3H, s), 2.68 (1H, dd,
J=12.5, 3.3 Hz), 2.57-0.94 (24H, m), 0.89-0.86 (21H), 0.59 (3H, s),
0.04 (3H, s), 0.03 (3H, s).
Reference Example 24
3-(Methoxymethyloxy)-22-(tert-butyldimethylsilyloxy)-7-oxa-cholest-8(9)-en-
-6-one (Compound P24)
[0295] Compound P23 (9.2 mg, 0.016 mmol) obtained in Reference
Example 23 was treated with acetic anhydride (1.5 mL) and sodium
acetate (100 mg, 1.22 mmol) in the same manner as Example 46 to
obtain the title compound (3.7 mg, 41%).
[0296] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66 (2H, m),
3.58 (1H, dd, J=8.0, 6.2 Hz), 3.52 (1H, m), 3.38 (3H, s), 2.39-1.00
(23H, m), 0.97 (3H, s), 0.89-0.86 (18H), 0.68 (3H, s), 0.034 (3H,
s), 0.028 (3H, s).
Reference Example 25
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-cholest-6,22-dien-24-one (Compound P25)
[0297] Compound P12 (5.40 g, 10.1 mmol) obtained in Reference
Example 12 and triphenylphosphine isobutanonmethylene (11.0 g, 31.8
mmol) were dissolved in anhydrous dimethyl sulfoxide (100 mL),
followed by stirring at 95.degree. C. for 72 hours. After being
left to cool, the reaction mixture was poured into water, followed
by extraction with ethyl acetate (200 mL.times.2). The organic
layer was washed with saturated brine, and dried over anhydrous
sodium sulfate. The residue (about 13 g) obtained by concentration
was purified by silica gel column chromatography (20 to 60% ethyl
acetate/n-hexane) to obtain the title compound (5.0 g, 82%).
[0298] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.15-8.11 (2H, m),
7.70-7.67 (2H, m), 6.69 (1H, dd, J=15.8, 8.8 Hz), 6.67 (1H, d,
J=8.6 Hz), 6.25 (1H, d, J=8.6 Hz), 6.05 (1H, d, J=15.8 Hz),
4.73-4.62 (2H, m), 4.10-3.96 (2H, m), 3.54 (1H, m), 3.34 (3H, s),
2.86 (1H, m), 2.28 (1H, m), 2.14-1.97 (3H, m), 1.88-1.25 (12H, m),
1.11 (3H, d, J=6.6 Hz), 1.09 (6H, d, J=7.0 Hz), 1.03 (3H, s), 0.87
(3H, s).
Reference Example 26
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-cholest-6-en-24-one (Compound P26) and
3-(methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl-
)-cholest-6-en-24-ol (Compound 27)
[0299] Compound P25 (4.95 g, 8.25 mmol) obtained in Reference
Example 25 was dissolved in methanol (50 mL), and nickel chloride
hexahydrate (250 mg, 1.05 mmol) was added thereto, followed by
stirring at 0.degree. C. for 10 minutes. Next, sodium borohydride
(811 mg, 21.4 mmol) was added in a small amount thereto, followed
by stirring at 0.degree. C. for 1 hour. Water was added to the
reaction mixture, followed by extraction with ethyl acetate, and
The organic layer was washed with saturated brine, and dried over
anhydrous sodium sulfate. The residue obtained by concentration was
purified by silica gel column chromatography (20 to 50% ethyl
acetate/n-hexane) to obtain compound P26 (944 mg, 19%) and compound
P27 (1.59 g, 32%).
[0300] Compound P26; .sup.1H NMR .delta.(ppm, CDCl.sub.3):
8.16-8.11 (2H, m), 7.72-7.65 (2H, m), 6.65 (1H, d, J=8.3 Hz), 6.26
(1H, d, J=8.3 Hz), 4.72-4.61 (2H, m), 4.06 (1H, dd, J=13.4, 4.4
Hz), 3.94 (1H, dd, J=12.1, 7.7 Hz), 3.54 (1H, m), 3.34 (3H, s),
2.60 (1H, m), 2.51-2.31 (2H, m), 2.12-1.22 (18H, m), 1.09 (6H, d,
J=7.0 Hz), 1.02 (3H, s), 0.92 (3H, d, J=6.2 Hz), 0.82 (3H, s).
[0301] Compound P27; .sup.1H NMR .delta.(ppm, CDCl.sub.3):
8.15-8.10 (2H, m), 7.71-7.64 (2H, m), 6.64 (1H, d, J=8.1 Hz), 6.26
(1H, d, J=8.1 Hz), 4.72-4.61 (2H, m), 4.05 (1H, dd, J=13.8, 4.0
Hz), 3.95 (1H, dd, J=11.0, 7.0 Hz), 3.53 (1H, m), 3.33 (3H, s),
3.28 (1H, m), 2.12-1.25 (22H, m), 1.01 (3H, s), 0.93-0.87 (9H),
0.82 (3H, s).
Reference Example 27
3-(Methoxymethyloxy)-cholest-5,7-dien-24-ol (Compound P28)
[0302] A mixture of Compounds P26 and P27 (3.78 g, 6.30 mmol)
obtained in Reference Example 26 were treated with lithium
aluminium hydride (2.50 g, 65.9 mmol) in the same manner as
Reference Example 13 to obtain the title compound (2.48 g,
89%).
[0303] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.53 (1H, m), 5.35
(1H, m), 4.67 (2H, s), 3.50 (1H, m), 3.34 (3H, s), 3.27 (1H, m),
2.48 (1H, m), 2.29 (1H, m), 2.08-1.16 (22H, m), 0.91 (3H, s),
0.94-0.85 (914), 0.59 (3H, s).
Reference Example 28
3-(Methoxymethyloxy)-cholest-7-en-6,24-diol (Compound P29)
[0304] Compound P28 (246.4 mg, 0.5950 mmol) obtained in Reference
Example 27 was treated with 1.0 mol/L of a borane-THF complex (3.0
mL, 3.0 mmol), 34.5% aqueous hydrogen peroxide (1.0 mL, 10 mmol)
and 6.0 mol/L of an aqueous solution of sodium hydroxide (1 mL, 6
mmol) in the same manner as Reference Example 2 to obtain the title
compound (145.0 mg, 53%).
[0305] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.15 (1H, br s),
4.69-4.64 (2H, m), 3.75 (1H, d, J=8.1 Hz), 3.46 (1H, m), 3.34 (3H,
s), 3.27 (1H, s), 2.28 (1H, m), 2.03-1.00 (25H, m), 0.92-0.85 (9H),
0.81 (3H, s), 0.52 (3H, s).
Reference Example 29
3-(Methoxymethyloxy)-cholest-7-en-6,24-dione (Compound P30)
[0306] Compound P29 (145.0 mg, 0.314 mmol) obtained in Reference
Example 28 was dissolved in dichloromethane, and Molecular Sieves 4
.ANG. (160 mg), N-methylmorpholine oxide (110 mg, 0.942 mmol) and a
small amount of tetrapropylammonium perruthenate were added
thereto, followed by stirring at room temperature for 2 hours. The
reaction mixture was filtered by using celite, and the filtrate was
concentrated, followed by purification by silica gel column
chromatography (10 to 30% ethyl acetate/n-hexane) to obtain the
title compound (108.2 mg, 75%).
[0307] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.67 (1H, s),
4.69-4.61 (2H, m), 3.48 (1H, m), 3.33 (3H, s), 2.56 (1H, m),
2.48-2.28 (2H, m), 2.24-1.17 (21H, m), 1.04 (6H, d, J=7.0 Hz), 0.89
(3H, d, J=6.1 Hz), 0.82 (3H, s), 0.55 (3H, s).
Reference Example 30
3-(Methoxymethyloxy)-6,8-seco-7-norcholest-8,24-dion-6-oic acid
(Compound P31)
[0308] Compound P30 (102.9 mg, 0.2250 mmol) obtained in Reference
Example 29 was treated with a small amount of ruthenium chloride
monohydrate, sodium periodate (150 mg, 0.701 mmol), cerium chloride
heptahydrate (15.0 mg, 0.0403 mmol) and lead acetate (80.0 mg,
0.180 mmol) in the same manner as Reference Example 5 to obtain the
title compound (17.7 mg, 17%).
[0309] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.66 (2H, s), 3.44
(1H, m), 3.35 (3H, s), 2.68-1.38 (25H, m), 1.23 (3H, s), 1.080 (6H,
d, J=6.8 Hz), 1.076 (3H, d, J=7.0 Hz), 0.58 (3H, s).
Reference Example 31
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-chol-6,23-diene-22-ol (Compound P32)
[0310] Compound P12 (13.2 g, 24.8 mmol) obtained in Reference
Example 12 was dissolved in anhydrous THF (50 mL) and 1 mol/L of a
vinylmagnesium bromide-THF solution (40 mL, 40 mmol) was slowly
added thereto dropwise under cooling at -78.degree. C. The reaction
mixture was stirred at -78.degree. C. for 1 hour, and then a
saturated aqueous ammonium chloride solution was added to increase
the temperature to room temperature, followed by extraction with
ethyl acetate (200 mL.times.2). The organic layer was washed with
saturated brine, and dried over anhydrous sodium sulfate. The
residue (about 12 g) obtained by concentration was purified by
silica gel column chromatography (20 to 80% ethyl acetate/n-hexane)
to obtain the title compound (10.3 g, 74%).
[0311] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.16-8.11 (2H, m),
7.73-7.66 (2H, m), 6.66 (1H, d, J=8.3 Hz), 6.28 (1H, d, J=8.3 Hz),
5.85 (1H, m), 5.24 (1H, d, J=17.2 Hz), 5.16 (1H, d, J=10.6 Hz),
4.73-4.62 (2H, m), 4.29 (1H, br s), 4.04 (2H, m), 3.54 (1H, m),
3.35 (3H, s), 2.15-1.40 (17H, m), 1.03 (3H, s), 0.89 (3H, d, J=6.8
Hz), 0.85 (3H, s).
Reference Example 32
Ethyl
3-(methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-
-diyl)-cholest-6,22-dien-26-oate (Compound P33)
[0312] Compound P32 (10.3 g, 18.4 mmol) obtained in Reference
Example 31, triethyl orthopropionate (35.0 mL, 174 mmol) and
propionic acid (1.50 mL, 20.1 mmol) were dissolved in toluene (75
mL) and Molecular Sieves 4 .ANG. (30 g) were added, followed by
stirring at 145.degree. C. for 2 hours. After being left to cool,
the reaction mixture was filtered, and water was added to the
filtrate, followed by extraction with ethyl acetate (150
mL.times.2). The organic layer was washed with a saturated aqueous
sodium bicarbonate solution and saturated brine, and dried over
anhydrous sodium sulfate, followed by purification by silica gel
column chromatography (20 to 60% ethyl acetate/n-hexane) to obtain
the title compound (8.4 g, 71%).
[0313] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.16-8.11 (2H, m),
7.71-7.66 (2H, m), 6.65 (1H, d, J=8.2 Hz), 6.26 (1H, d, J=8.2 Hz),
5.33-5.22 (2H, m), 4.72-4.63 (2H, m), 4.10 (2H, q, J=7.1 Hz), 4.07
(1H, m), 3.95 (1H, dd, J=12.0, 7.0 Hz), 3.55 (1H, m), 3.35 (3H, s),
2.47-1.27 (19H, m), 1.24 (3H, t, J=7.1 Hz), 1.11 (3H, d, J=6.9 Hz),
1.03 (3H, s), 1.01 (3H, d, J=6.6 Hz), 0.84 (3H, s).
Reference Example 33
Ethyl
3-(methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-
-diyl)-cholest-6-en-26-oate (Compound P34)
[0314] Compound P33 (4.00 g, 6.21 mmol) obtained in Reference
Example 32 was dissolved in ethyl acetate (80 mL), and then 10%
palladium-carbon powders (400 mg) were added thereto, followed by
stirring at room temperature for 3 hours under a hydrogen
atmosphere. The catalyst was separated by filtration, and the
filtrate was concentrated to obtain the title compound.
[0315] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.16-8.11 (2H, m),
7.72-7.65 (2H, m), 6.65 (1H, d, J=8.1 Hz), 6.27 (1H, d, J=8.1 Hz),
4.73-4.62 (2H, m), 4.10 (2H, q, J=7.2 Hz), 4.07 (1H, m), 3.93 (1H,
m), 3.54 (1H, m), 3.34 (3H, s), 2.48-1.28 (23H, m), 1.25 (3H, t,
J=7.2 Hz), 1.10 (3H, d, J=7.2 Hz), 1.02 (3H, s), 1.00 (3H, d, J=7.2
Hz), 0.83 (3H, s).
Reference Example 34
3-(Methoxymethyloxy)-cholest-5,7-dien-26-ol (Compound P35)
[0316] Compound P34 obtained in Reference Example 33 was treated
with lithium aluminium hydride (3.10 g, 81.7 mmol) in the same
manner as Reference Example 13 to obtain the title compound (1.53
g, 56% (2 steps)).
[0317] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, m), 5.38
(1H, m), 4.71 (2H, s), 3.52 (2H, m), 3.44 (1H, m), 3.38 (3H, s),
2.52 (1H, m), 2.25 (1, m), 2.06-1.25 (24H, m), 1.03 (3H, d, J=6.6
Hz), 0.94 (3H, s), 0.92 (3H, d, J=6.4 Hz), 0.63 (3H, s).
Reference Example 35
3-(Methoxymethyloxy)-cholest-7-en-6,26-diol (Compound P36)
[0318] Compound P35 (1.53 g, 3.45 mmol) obtained in Reference
Example 34 was treated with 1.0 mol/L of a borane-THF complex/THF
solution (16 mL, 16 mmol), 6.0 mol/L of an aqueous solution of
sodium hydroxide (5.80 mL, 34.5 mmol) and 30% aqueous hydrogen
peroxide (3.90 mL, 34.5 mmol) in the same manner as Reference
Example 2 to obtain the title compound (690 mg, 43%).
[0319] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.17 (1H, br s),
4.72-4.66 (2H, m), 3.78 (1H, br d, J=8.3 Hz), 3.52-3.39 (3H, m),
3.36 (3H, s), 2.30 (1H, m), 2.02 (1H, m), 1.80-1.04 (26H, m), 0.91
(3H, d, J=6.6 Hz), 0.90 (3H, d, J=6.6 Hz), 0.83 (3H, s), 0.53 (3H,
s).
Reference Example 36
3-(Methoxymethyloxy)-cholest-7-en-6-on-26-ol (Compound P37)
[0320] Compound P36 (690 mg, 1.49 mmol) obtained in Reference
Example 35 was oxidized with manganese oxide (5 g, 57.5 mmol) in
the same manner as Reference Example 8 to obtain the title compound
(380 mg, 55%).
[0321] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, br s),
4.74-4.66 (2H, m), 3.61-3.42 (3H, m), 3.38 (3H, s), 2.29-1.25 (27H,
m), 0.93 (3H, d, J=7.0 Hz), 0.91 (3H, d, J=7.0 Hz), 0.86 (3H, s),
0.59 (3H, s).
Reference Example 37
3-(Methoxymethyloxy)-26-(triisopropylsilyloxy)-cholest-7-en-6-one
(Compound P38)
[0322] Imidazole (190 mg, 2.79 mmol) and triisopropylsilane
chloride (0.30 mL, 1.40 mmol) were added to an anhydrous DMF
solution (6 mL) containing compound P37 (290 mg, 0.630 mmol)
obtained in Reference Example 36, followed by stirring at room
temperature for 2 hours. The reaction mixture was poured into a
saturated aqueous ammonium chloride solution, followed by
extraction with ethyl acetate (50 mL.times.2). The organic layer
was washed with water and saturated brine, and dried over anhydrous
sodium sulfate. The residue obtained by concentration was purified
by silica gel column chromatography (10 to 40% ethyl
acetate/n-hexane) to obtain the title compound (381 mg, 98%).
[0323] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, br s),
4.74-4.66 (2H, m), 3.56-3.43 (3H, m), 3.38 (3H, s), 2.24-1.25 (26H,
m), 1.06-1.05 (21H, m), 0.93 (3H, d, J=6.6 Hz), 0.88 (3H, d, J=6.6
Hz), 0.87 (3H, s), 0.60 (3H, s).
Reference Example 38
3-(Methoxymethyloxy)-26-(triisopropylsilyloxy)-cholest-6-on-7,8-diol
(Compound P39)
[0324] Ethyl acetate (1 mL) and acetonitrile (3 mL) were
sequentially added to an aqueous solution (1 mL) containing sodium
periodate (51.8 mg, 0.242 mmol) and cerium chloride heptahydrate
(36.1 mg, 0.0969 mmol). Subsequently, 0.1 mol/L of an aqueous
solution of ruthenium chloride (0.21 mL, 0.021 mmol) was added
thereto, followed by stirring at 0.degree. C. for 10 minutes. An
ethyl acetate solution (2 mL) of compound P37 (49.2 mg, 0.0799
mmol) obtained in Reference Example 36 was added to the mixture at
0.degree. C., followed by stirring at room temperature for 1 hour.
Anhydrous sodium sulfate (10 g) was added to the reaction mixture,
followed by filtration, and the filtrate was washed with a
saturated aqueous sodium sulfite solution and saturated brine, and
dried over anhydrous sodium sulfate, followed by purification by
silica gel column chromatography (10 to 30% ethyl acetate/n-hexane)
to obtain the title compound (37.2 mg, 72%).
[0325] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.70-4.65 (2H, m),
4.26 (1H, s), 3.56-3.41 (3H, m), 3.36 (3H, s), 2.50-1.09 (28H, m),
1.05-1.03 (21H, m), 0.89 (3H, d, J=6.4 Hz), 0.86 (3H, s), 0.84 (3H,
d, J=7.0 Hz), 0.59 (3H, s).
Reference Example 39
3-(Methoxymethyloxy)-26-(triisopropylsilyloxy)-6,8-seco-7-norcholest-8-on--
6-oic acid (Compound P40)
[0326] Potassium carbonate (106.5 mg, 0.7710 mmol) and lead (IV)
tetraacetate (138.6 mg, 0.3130 mmol) were added to an hydrous
toluene solution (10 mL) containing compound P39 (40.7 mg, 0.0626
mmol) obtained in Reference Example 38 at 0.degree. C.
[0327] The mixture was stirred at room temperature for 20 minutes,
and then sodium sulfate (10 g) was added, followed by filtration.
The filtrate was concentrated, followed by purification by silica
gel preparative thin layer chromatography (n-hexane-ethyl acetate
(3:1)) to obtain
3-(methoxymethyloxy)-26-(triisopropylsilyloxy)-6,8-seco-7-norchole-
st-8-on-6-oic acid (7.9 mg, 20%) and
3-(methoxymethyloxy)-26-(triisopropylsilyloxy)-7-oxa-cholest-8(9)-en-6-on-
e (2.5 mg, 6.5%).
3-(methoxymethyloxy)-26-(triisopropylsilyloxy)-6,8-seco-7-norcholest-8-on-
-6-oic acid;
[0328] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s),
3.54-3.42 (3H, m), 3.36 (3H, s), 2.67-1.35 (27H, m), 1.24 (3H, s),
1.06-1.04 (21H, m), 0.92 (3H, d, J=6.8 Hz), 0.88 (3H, d, J=6.8 Hz),
0.58 (3H, s).
Reference Example 40
3-(Methoxymethyloxy)-26-(triisopropylsilyloxy)-7-oxa-cholest-8(9)-en-6-one
(Compound P41)
[0329] Pyridine (100 .mu.L) and thionyl chloride (50 .mu.L) were
added to an anhydrous dichloromethane solution (2 mL) containing
compound P40 (11.8 mg, 0.0186 mmol) obtained in Reference Example
39 at 0.degree. C., followed by stirring for 15 minutes. The
reaction mixture was poured into a saturated ammonium chloride
solution, followed by extraction with chloroform. The organic layer
was washed with saturated brine, and dried over anhydrous sodium
sulfate, followed by purification by silica gel preparative thin
layer chromatography (n-hexane-ethyl acetate (3:1)) to obtain the
title compound (9.1 mg, 79%).
[0330] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66 (2H, m),
3.54-3.43 (3H, m), 3.38 (3H, s), 2.34-1.25 (25H, m), 1.05-1.03
(21H, m), 0.97 (3H, s), 0.93 (3H, d, J=6.2 Hz), 0.88 (3H, d, J=6.8
Hz), 0.67 (3H, s).
Reference Example 41
3-(Methoxymethyloxy)-20-(hydroxymethyl)-pregna-7-en-6-ol (Compound
P42)
[0331] Compound P13 (340 mg, 0.909 mg) obtained in Reference
Example 13 was treated with 1.0 mol/L of a borane-THF complex (3.6
mL, 3.6 mmol), 6.0 mol/L of an aqueous solution of sodium hydroxide
(1.5 mL, 9.0 mmol) and 34.5% aqueous hydrogen peroxide (1.0 mL, 10
mmol) in the same manner as Reference Example 2 to obtain the title
compound (310 mg, 87%).
[0332] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.19 (1H, br s), 4.71
(2H, m), 3.81 (1H, m), 3.65 (1H, m), 3.50 (1H, m), 3.39 (1H, m),
3.38 (3H, s), 2.31 (1H, m), 2.03 (1H, m), 1.92-1.13 (19H, m), 1.06
(3H, d, J=6.6 Hz), 0.85 (3H, s), 0.57 (3H, s).
Reference Example 42
3-(Methoxymethyloxy)-20-carboxy-pregna-7-en-6-on-22-oic acid
(Compound P43)
[0333] Compound P42 (141.7 mg, 0.3610 mmol) obtained in Reference
Example 41 was dissolved in acetone (10 mL), and then Jones reagent
[prepared by dissolving 26.7 g of chromium (VI) oxide in 23 mL of
conc. sulfuric acid-40 mL of water, and diluting with water to be
100 mL of whole volume] was slowly added dropwise until the
solution turned limegreen color under ice-cooling. After stirring
at 0.degree. C. for 30 minutes, isopropanol (about 2 mL) was added
to decompose the excess of the oxidizing agent. The reaction
mixture was diluted with water, followed by extraction with ethyl
acetate. The organic layer was washed with saturated brine, dried
over anhydrous sodium sulfate, and concentrated to yield a residue
(about 110 mg), to which a mixed solvent of ethyl acetate/n-hexane
was added for crystallization to obtain the title compound (91.6
mg, 63%).
[0334] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.73 (1H, s), 4.71
(2H, m), 3.53 (1H, m), 3.38 (3H, s), 2.46 (1H, m), 2.30-1.37 (19H,
m), 1.28 (3H, d, J=6.8 Hz), 0.87 (3H, s), 0.63 (3H, s).
Reference Example 43
tert-Butyl 3-(methoxymethyloxy)-20-carboxy-pregna-7-en-6-on-22-oate
(Compound P44)
[0335] Compound P43 (148.9 mg, 0.3690 mmol) obtained in Reference
Example 42 and N,N-dimethylformamide tert-butyl acetal (707 .mu.L,
2.95 mmol) were dissolved in toluene (2 mL), followed by stirring
at 80.degree. C. for 2 hours. The reaction mixture was
concentrated, followed by purification by silica gel column
chromatography (10 to 30% ethyl acetate/n-hexane) to obtain the
title compound (126.8 mg, 75%).
[0336] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.67 (1H, br s), 4.67
(2H, m), 3.50 (1H, m), 3.35 (3H, s), 2.29-2.01 (6H, m), 1.84-1.25
(13H, m), 1.41 (9H, s), 1.14 (3H, d, J=6.8 Hz), 0.84 (3H, s), 0.58
(3H, s).
Reference Example 44
tert-Butyl
3-(methoxymethyloxy)-20-carboxy-6,8-seco-7-norpregna-8-on-6-oat- e
(Compound P45)
[0337] Compound P44 (80.8 mg, 0.176 mmol) obtained in Reference
Example 43 was dissolved in acetic acid (3 mL), and lead acetate
(330 mg, 0.744 mmol) and a small amount of ruthenium chloride
hydrate were added thereto, followed by stirring at room
temperature for 1 hour. Isopropanol (2 mL) was added to the
reaction mixture, followed by filtration using celite, and the
filtrate was extracted with ethyl acetate. The organic layer was
sequentially washed with a saturated aqueous sodium sulfite
solution and saturated brine, and dried over anhydrous sodium
sulfate. The residue obtained by concentration was purified by
silica gel column chromatography (10 to 50% ethyl acetate/n-hexane)
to obtain the title compound (30.1 mg, 36%).
[0338] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.66 (2H, m), 3.44
(1H, m), 3.35 (3H, s), 2.65 (1H, dd, J=12.8, 3.3 Hz), 2.57-1.25
(19H, m), 1.42 (9H, s), 1.23 (3H, s), 1.15 (3H, d, J=6.8 Hz), 0.58
(3H, s).
Reference Example 45
3-(Acetyloxy)-ergosta-5,7,22-triene (Compound P46)
[0339] Ergosterol (5.00 g, 12.6 mmol) was dissolved in
dichloromethane (100 mL), and pyridine (4 mL) and acetyl chloride
(2 mL) were sequentially added at 0.degree. C. thereto, followed by
stirring at room temperature for 18 hours. The reaction mixture was
concentrated to yield a residue and then methanol was added to the
obtained residue for trituration to obtain the title compound (5.21
g, 94%).
[0340] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, dd, J=6.1,
2.4 Hz), 5.38 (1H, m), 5.13-5.27 (2H, m), 4.70 (1H, m), 2.50 (1H,
m), 2.36 (1H, m), 2.04 (3H, s), 2.05-1.25 (18H, m), 1.04 (3H, d,
J=6.6 Hz), 0.95 (3H, s), 0.92 (3H, d, J=7.0 Hz), 0.84 (3H, d, J=6.8
Hz), 0.82 (3H, d, J=6.8 Hz), 0.63 (3H, s), 5.57 (1H, dd, J=6.1, 2.4
Hz), 5.38 (1H, m), 5.13-5.27 (2H, m), 4.70 (1H, m), 2.50 (1H, m),
2.36 (1H, m), 2.04 (3H, s), 2.05-1.25 (18H, m), 1.04 (3H, d, J=6.6
Hz), 0.95 (3H, s), 0.92 (3H, d, J=7.0 Hz), 0.84 (3H, d, J=6.8 Hz),
0.82 (3H, d, J=6.8 Hz), 0.63 (3H, s).
Reference Example 46
3-(Acetyloxy)-ergosta-7,22-dien-6-ol (Compound P47)
[0341] Compound P46 (500 mg, 1.14 mmol) obtained in Reference
Example 45 was dissolved in THF (13 mL), and then 1.0 mol/L of a
borane-THF complex (2.3 mL, 2.3 mmol) was added dropwise under
ice-cooling, followed by stirring at room temperature for 16 hours.
The reaction mixture was again ice-cooled, and then water (4 mL),
1.0 mol/L of an aqueous solution of sodium hydroxide (4 mL) and
34.5% aqueous hydrogen peroxide (2.0 mL) were sequentially added
dropwise, followed by stirring at room temperature for 2.5 hours.
Water was added to the reaction mixture, followed by extraction
with ethyl acetate (40 mL.times.2). The organic layer was washed
with saturated brine, dried over anhydrous sodium sulfate, and
concentrated to yield a residue (600 mg), which was purified by
silica gel column chromatography (20 to 40% ethyl acetate/n-hexane)
to obtain the title compound (120 mg, 23%).
[0342] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.26-5.12 (3H, m),
4.70 (1H, m), 3.79 (1H, d, J=9.4 Hz), 2.30 (1H, m), 2.03 (3H, s),
1.86-1.25 (21H, m), 1.02 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.8 Hz),
0.86 (3H, s), 0.84 (3H, d, J=6.8 Hz), 0.82 (3H, d, J=6.6 Hz), 0.55
(3H, s).
Reference Example 47
3-(Acetyloxy)-ergosta-7,22-dien-6-one (Compound P48)
[0343] Compound P47 (170 mg, 0.373 mmol) obtained in Reference
Example 46 was dissolved in dichloromethane (30 mL), and then
manganese (IV) oxide (1.25 g, 14.4 mmol) was added thereto,
followed by stirring at room temperature for 17 hours. After the
mixture was filtrated, the filtrate was concentrated to yield a
residue (270 mg), which was purified by silica gel column
chromatography (10 to 20% ethyl acetate/n-hexane) to obtain the
title compound (108 mg, 64%).
[0344] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.71 (1H, s),
5.27-5.11 (2H, m), 4.71 (1H, m), 2.42-1.10 (21H, m), 2.03 (3H, s),
1.03 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.8 Hz), 0.87 (3H, s), 0.84
(3H, d, J=6.6 Hz), 0.83 (3H, d, J=6.6 Hz), 0.60 (3H, s).
Reference Example 48
3-(Acetyloxy)-ergosta-7-en-6-one (Compound P49)
[0345] The compound P48 (45.5 mg, 0.100 mmol) obtained in Reference
Example 47 was dissolved in ethyl acetate (5 mL), and then 10%
palladium-carbon (48 mg) was added, followed by stirring at room
temperature for 1 hour under a hydrogen atmosphere. After
filtration of the mixture, the filtrate was concentrated to yield a
residue. The residue was purified by silica gel column
chromatography (5 to 10% ethyl acetate/n-hexane) to obtain the
title compound (34.6 mg, 76%).
[0346] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.71 (1H, s), 4.71
(1H, m), 2.29 (1H, dd, J=12.3, 3.7 Hz), 2.23-1.20 (24H, m), 2.02
(3H, s), 0.93 (3H, d, J=6.8 Hz), 0.86 (3H, s), 0.84 (3H, d, J=7.0
Hz), 0.77 (6H, d, J=6.8 Hz), 0.58 (3H, s).
Reference Example 49
3-(Acetyloxy)-6,8-seco-7-norergosta-8-on-6-oic acid (Compound
P50)
[0347] Compound P49 (172.9 mg, 0.3790 mmol) obtained in Reference
Example 48 was dissolved in a mixed solvent of chloroform (2
mL)-acetonitrile (1 mL)-water (1 mL), and then sodium periodate
(350 mg, 1.64 mmol) and a small amount of ruthenium chloride
hydrate were added thereto, followed by stirring at room
temperature for 2.5 hours. Isopropanol (3 mL) was added to the
reaction mixture and stirred, followed by filtration using celite.
Water was added to the filtrate, followed by extraction with
chloroform (25 mL.times.2). The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated to yield a residue (220 mg). The residue was dissolved
in toluene (5 mL), and lead acetate (210 mg, 0.474 mmol) was added
thereto, followed by stirring at room temperature for 6 hours. The
reaction mixture was filtrated by using celite, and water was added
to the filtrate, followed by extraction with chloroform (25
ml.times.2). The organic layer was washed with saturated brine and
dried over anhydrous magnesium sulfate, followed by purification by
silica gel column chromatography (10 to 20% ethyl acetate/n-hexane)
to obtain the title compound (24.0 mg, 13%).
[0348] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.62 (1H, m), 2.70
(1H, dd, J=12.7, 3.3 Hz), 2.58-2.44 (2H, m), 2.33-2.29 (2H, m),
2.16-1.05 (21H, m), 2.02 (3H, s), 1.24 (3H, s), 0.94 (3H, d, J=6.0
Hz), 0.85 (3H, d, J=6.8 Hz), 0.77 (6H, d, J=6.6 Hz), 0.58 (3H,
s).
Reference Example 50
(3S,10R,13R,14R,17R)-3-Methoxymethoxy-17-[(S)-1-methoxypropan-2-yl]-10,13--
dimethyl-2,3,4,9,10,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phena-
nthrene (Compound P51)
[0349] Compound P13 (5.51 g, 14.7 mmol) obtained in Reference
Example 13 and iodomethane (60.0 mL, 958 mmol) were dissolved in
DMF (150 mL), and sodium hydride (4.95 g, 124 mmol) was added
thereto, followed by stirring at room temperature for 2.5 hours.
The solution was cooled to 0.degree. C., and a saturated aqueous
ammonium chloride solution was added dropwise, followed by
extraction with diethyl ether (100 mL.times.2). The organic layer
was washed with saturated brine, dried over anhydrous sodium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (10 to 30% ethyl
acetate/n-hexane) to obtain the title compound (5.72 g, 100%).
[0350] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, m), 5.38
(1H, m), 4.70 (2H, s), 3.53 (1H, m), 3.40 (1H, m), 3.38 (3H, s),
3.32 (3H, s), 3.13 (1H, dd, J=7.3 Hz, 16.1 Hz), 2.52 (1H, m), 2.33
(1H, m), 2.10-1.20 (16H, m), 1.06 (3H, d, J=6.6 Hz), 0.94 (3H, s),
0.63 (3H, s).
Reference Example 51
(3S,5S,6S,10R,13R,14R,17R)-3-Methoxymethoxy-17-[(S)-1-methoxypropan-2-yl]--
10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclo-
penta[a]phenanthren-6-ol (Compound P52)
[0351] Compound P51 (5.72 g, 14.7 mmol) obtained in Reference
Example 50 was dissolved in THF (150 mL), and then 1.0 mol/L of a
borane-THF complex (44.1 mL, 44.1 mmol) was added dropwise thereto
under ice-cooling, followed by stirring at room temperature for 1
hour. The reaction mixture was again ice-cooled, and then water (15
mL), 34.5% aqueous hydrogen peroxide (10 mL) and 1.0 mol/L of an
aqueous solution of sodium hydroxide (10 mL) were sequentially
added dropwise, followed by stirring at room temperature for 1
hour. Water was added to the reaction mixture, followed by
extraction with ethyl acetate (150 mL.times.3). The organic layer
was washed with saturated brine, dried over anhydrous magnesium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (20 to 40% ethyl
acetate/n-hexane) to obtain the title compound (5.19 g, 87%).
[0352] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.18 (1H, m),
4.72-4.68 (2H, m), 3.79 (1H, br s), 3.49 (1H, m), 3.38 (3H, s),
3.33 (1H, m), 3.31 (3H, s), 3.13 (1H, dd, J=7.1 Hz, 16.1 Hz), 2.30
(1H, m), 2.00-1.17 (29H, m), 1.03 (3H, d, J=6.6 Hz), 0.84 (3H, s),
0.56 (3H, s).
Reference Example 52
(3S,5S,10R,13R,14R,17R)-3-Methoxymethoxy-17-[(S)-1-methoxypropan-2-yl]-10,-
13-dimethyl-2,3,4,5,9,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phe-
nanthren-6(10H)-one (Compound P53)
[0353] Compound P52 (5.19 g, 12.8 mmol) obtained in Reference
Example 51 was dissolved in dichloromethane (128 mL), and then
N-methylmorpholine oxide (4.49 g, 38.3 mmol) and
tetrapropylammonium perruthenate (225 mg, 0.64 mmol) were added
thereto, followed by stirring at room temperature for 30 minutes.
The reaction mixture was filtered by using celite, and the filtrate
was concentrated, followed by purification by silica gel column
chromatography (10 to 30% ethyl acetate/n-hexane) to obtain the
tilte compound (4.2 g, 70%).
[0354] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, m),
4.72-4.68 (2H, m), 3.53 (1H, m), 3.38 (3H, s), 3.32 (1H, m), 3.32
(3H, s), 3.16 (1H, dd, J=6.8 Hz, J=9.0 Hz), 2.30-1.30 (19H, m),
1.06 (3H, d, J=6.6 Hz), 0.87 (3H, s), 0.61 (3H, s).
Reference Example 53
(3S,5S,7S,8S,10R,13R,14R,17R)-7,8-Dihydroxy-3-methoxymethoxy-17-[(S)-1-met-
hoxypropan-2-yl]-10,13-dimethyltetradecahydro-1H-cyclopenta[a]phenanthren--
6(10H)-one (Compound P54)
[0355] Compound P53 (1.79 g, 4.09 mmol) obtained in Reference
Example 52, sodium periodate (5.24 g, 24.5 mmol) and cerium
chloride heptahydrate (1.52 g, 4.09 mmol) were dissolved in a mixed
solvent of acetonitrile (200 mL)-ethyl acetate (200 mL)-water (67
mL), and then ruthenium trichloride (254 mg, 1.23 mmol) was added
thereto, followed by stirring at 0.degree. C. for 30 minutes. A
saturated aqueous sodium thiosulfate solution was added to the
reaction mixture, followed by extraction with ethyl acetate (500
mL.times.3). The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated to obtain
the title compound (1.25 g, 65%).
[0356] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.71-4.67 (2H, m),
4.26 (1H, br s), 3.55 (1H, m), 3.47 (1H, br s), 3.39-3.30 (7H, m),
3.11 (1H, dd, J=7.2 Hz, 9.0 Hz), 2.49-1.21 (20H, m), 1.03 (3H, d,
J=6.6 Hz), 0.91 (3H, s), 0.84 (3H, s).
Reference Example 54
(1S,2R,5S)-5-Methoxymethoxy-2{(1R,3aR,7aR)-1-[(S)-1-methoxypropan-2-yl]-7a-
-methyl-4-oxooctahydro-1H-inden-5-yl}-2-methylcyclohexanecarboxylic
acid (Compound P55)
[0357] Compound P54 (1.25 g, 2.65 mmol) obtained in Reference
Example 53 was dissolved in pyridine (50 mL), and lead tetraacetate
(2.35 g, 5.30 mmol) was added thereto, followed by stirring at
0.degree. C. for 30 minutes. A saturated aqueous sodium thiosulfate
solution was added to the reaction mixture, followed by extraction
with ethyl acetate (50 mL.times.3). The organic layer was washed
with saturated sodium thiosulfate, and dried over anhydrous
magnesium sulfate, followed by purification by silica gel column
chromatography (20 to 33% ethyl acetate/n-hexane) to obtain the
title compound (1.47 g).
[0358] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s), 3.36
(3H, s), 3.31 (3H, s), 3.30 (1H, m), 3.13 (1H, dd, J=6.4 Hz, 15.6
Hz), 2.67 (1H, m), 2.58-1.35 (20H, m), 1.25 (3H, s), 1.04 (3H, d,
J=6.2 Hz), 0.60 (3H, s).
Reference Example 55
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-pregna-6-en-22-ol (Compound P56)
[0359] Compound P12 (23.9 g, 44.8 mmol) obtained in Reference
Example 12 was dissolved in a mixed solvent of THF (450 mL) and
water (50 mL), and cooled to 0.degree. C., and then sodium
borohydride (1.69 g, 44.8 mmol) was added thereto, followed by
stirring at 30 minutes. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with ethyl acetate (300 mL.times.3). The organic layer was washed
with saturated brine, and dried over anhydrous magnesium sulfate.
The residue obtained by concentration was purified by silica gel
column chromatography (30 to 50% ethyl acetate/n-hexane) to obtain
the title compound (20.8 g, 87%).
[0360] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.17-8.13 (2H, m),
7.71-7.68 (2H, m), 6.66 (1H, d, J=8.1 Hz), 6.30 (1H, d, J=8.3 Hz),
4.70-4.65 (2H, m), 4.10-3.93 (2H, m), 3.66 (1H, dd, J=3.1 Hz, 10.6
Hz), 3.56 (1H, m), 3.38 (1H, m), 3.35 (3H, s), 2.14-1.23 (17H, m),
1.06 (3H, d, J=6.6 Hz), 1.03 (3H, s), 0.86 (3H, s).
Reference Example 56
3-(methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-pregna-6-en-22-methallyl ether (Compound P57)
[0361] Compound P56 (9.67 g, 18.1 mmol) obtained in Reference
Example 55, methallyl bromide (9.00 mL, 90.5 mmol) and
tetrabutylammonium iodide (6.70 g, 18.1 mmol) were dissolved in DMF
(60 mL), and sodium hydride (2.20 g, 54.3 mmol) was added thereto,
followed by stirring at room temperature for 4 hours. The mixture
was cooled to 0.degree. C., and then a saturated aqueous ammonium
chloride solution was added to the reaction mixture, followed by
extraction with diethyl ether (100 mL.times.3). The organic layer
was washed with saturated brine, and dried over anhydrous magnesium
sulfate. The residue obtained by concentration was purified by
silica gel column chromatography (20 to 40% ethyl acetate/n-hexane)
to obtain the title compound (3.26 g, 93%).
[0362] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.17-8.13 (2H, m),
7.72-7.68 (2H, m), 6.66 (1H, d, J=8.3 Hz), 6.27 (1H, d, J=8.4 Hz),
3.55 (1H, m), 3.35 (3H, s), 1.06 (3H, d, J=6.6 Hz), 1.03 (3H, s),
0.85 (3H, s).
Reference Example 57
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-pregna-6-en-22-isobutyl ether (Compound P58)
[0363] Compound P57 (3.26 g, 5.06 mmol) obtained in Reference
Example 56 was dissolved in ethyl acetate (20 mL), and then 10%
palladium-carbon (326 mg) was added thereto, followed by stirring
at room temperature for 9 hours under a hydrogen atmosphere. The
catalyst was separated by filtration, and the filtrate was
concentrated to obtain the title compound (2.99 g, 85%).
[0364] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.15-8.12 (2H, m),
7.71-7.68 (2H, m), 6.66 (1H, d, J=7.9 Hz), 6.27 (1H, d, J=8.1 Hz),
4.70-4.67 (2H, m), 4.13-3.93 (2H, m), 3.55 (1H, m), 3.40-3.35 (5H,
m), 3.20-3.05 (2H, m), 2.14-1.20 (17H, m), 1.04 (3H, d, J=5.3 Hz),
1.03 (3H, s), 0.90 (6H, dd, J=3.1 Hz, 6.8 Hz), 0.85 (3H, s).
Reference Example 58
(10R,13R,14R,17R)-17-(2-Isobutoxyethyl)-3-methoxymethoxy-10,13-dimethyl-2,-
3,4,9,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene
(Compound P59)
[0365] Compound P58 (2.99 g, 5.06 mmol) obtained in Reference
Example 57 was treated with lithium aluminium hydride (960 mg, 25.3
mmol) in the same manner as Reference Example 13 to obtain the
title compound (1.31 g, 60%).
[0366] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, m), 5.28
(1H, m), 4.71 (2H, s), 3.53 (1H, m), 3.37 (1H, m), 3.38 (3H, s),
3.20 (1H, dd, J=6.6 Hz, 9.4 Hz), 3.11-3.07 (2H, m), 2.52 (1H, m),
2.37-1.20 (18H, m), 1.06 (3H, d, J=6.4 Hz), 0.94 (3H, s), 0.90 (6H,
dd, J=2.8 Hz, 6.6 Hz), 0.63 (3H, s).
Reference Example 59
(5S,6S,10R,13R,14R,17R)-17-(2-Isobutoxyethyl)-3-methoxymethoxy-10,13-dimet-
hyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phe-
nanthren-6-ol (Compound P60)
[0367] Compound P59 (1.31 g, 3.03 mmol) obtained in Reference
Example 58 was treated with 1.0 mol/L of a borane-THF complex (9.0
mL, 9.0 mmol) in the same manner as Reference Example 2 to obtain
the title compound (0.73 g, yield 54%).
[0368] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.18 (1H, m),
4.74-4.68 (2H, m), 3.80 (1H, br s), 3.50 (1H, m), 3.38 (1H, m),
3.38 (3H, s), 3.19 (1H, dd, J=6.6 Hz, 9.2 Hz), 3.11-3.07 (2H, m),
2.33 (1H, m), 2.06-1.20 (20H, m), 1.04 (3H, d, J=6.4 Hz), 0.84 (3H,
s) 0.89 (6H, dd, J=2.8 Hz, 6.8 Hz), 0.56 (3H, s).
Reference Example 60
(5R,10R,13R,14R,17R)-17-(2-Isobutoxyethyl)-3-methoxymethoxy-10,13-dimethyl-
-2,3,4,5,9,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-6(-
10H)-one (Compound P61)
[0369] Compound P60 (2.63 g, 5.86 mmol) obtained in Reference
Example 59 was treated with N-methylmorpholine oxide (2.10 g, 17.6
mmol) and tetrapropylammonium perruthenate (103 mg, 0.293 mmol) in
the same manner as Reference Example 4 to obtain the title compound
(2.6 g, 100%).
[0370] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, s),
4.72-4.68 (2H, m), 3.52 (1H, m), 3.38 (3H, s), 3.39-3.06 (4H, m),
2.29-1.20 (22H, m), 1.05 (3H, d, J=6.6 Hz), 0.91-0.89 (6H, m), 0.87
(3H, s), 0.62 (3H, s).
Reference Example 61
(5S,7S,8S,10R,13R,14R,17R)-7,8-Dihydroxy-17-(2-isobutyloxyethyl)-3-methoxy-
methoxy-10,13-dimethyl
tetradecahydro-1H-cyclopenta[a]phenanthren-6(10H)-one (Compound
P62)
[0371] Compound P61 (2.62 g, 5.86 mmol) obtained in Reference
Example 60 was treated with sodium periodate (7.50 g, 35.2 mmol),
cerium chloride heptahydrate (2.18 g, 5.86 mmol) and ruthenium
chloride (365 mg, 1.76 mmol) in the same manner as Reference
Example 5 to obtain the title compound (2.03 g, 72%).
[0372] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.67 (2H, m),
4.27 (1H, s), 3.71-3.04 (8H, m), 2.47-2.37 (2H, m), 2.05-0.83 (35H,
m).
Reference Example 62
(1S,2R)-2-[(1R,3aR,7aR)-1-(2-Isobutoxyethyl)-7a-methyl-4-oxooctahydro-1H-i-
nden-5-yl]-5-methoxymethoxy-2-methylcyclohexanecarboxylic acid
(Compound P63)
[0373] Compound P62 (2.03 g) obtained in Reference Example 61 was
treated with lead tetraacetate (3.70 g, 8.45 mmol) in the same
manner as Reference Example 39 to obtain a crude product (2.34 g)
of the title compound.
Reference Example 63
(3S,5S,6S,10R,13R,14R)-3-Methoxymethoxy-10,13-dimethyl-17-[(S)-1-triisopro-
pylsilyloxy
propan-2-yl]-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclop-
enta[a]phenanthren-6-ol (Compound P82)
[0374] Compound P13 (3.66 g, 9.78 mmol) obtained in Reference
Example 13 was dissolved in DMF (90 mL), and imidazole (1.99 g,
29.3 mmol), triisopropylsilyl chloride (4.20 mL, 19.6 mmol) and
catalytic amount of 4-dimethylamino pyridine were added thereto,
followed by stirring at room temperature for 12 hours. Water was
added to the reaction mixture, followed by extraction with diethyl
ether (100 mL.times.3). The organic layer was washed with saturated
brine, dried over anhydrous magnesium sulfate, and concentrated.
The resulting residue (5.3 g) was dissolved in THF (100 mL), and
1.0 mol/L of a borane-THF complex (30.0 mL, 30.0 mmol) was added
dropwise thereto under ice-cooling, followed by stirring at room
temperature for 1 hour. The reaction mixture was again ice-cooled,
and then water (15 mL), 34.5% aqueous hydrogen peroxide (10.0 mL)
and 1.0 mol/L of an aqueous solution of sodium hydroxide (10 mL)
were sequentially added dropwise, followed by stirring at room
temperature for 1 hour. Water was added to the reaction mixture,
followed by extraction with ethyl acetate (100 mL.times.3). The
organic layer was washed with saturated sodium thiosulfate and
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated to yield a residue. The residue was purified by silica
gel column chromatography (20 to 40% ethyl acetate/n-hexane) to
obtain the title compound (2.63 g, 49% (2 steps)).
[0375] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.18 (1H, m),
4.72-4.67 (2H, m), 3.80 (1H, br s), 3.67 (1H, dd, J=3.3 Hz, 9.4
Hz), 3.50 (1H, m), 3.38 (1H, m), 3.38 (3H, s), 2.31 (1H, m), 2.04
(1H, m), 1.90-1.19 (18H, m) 1.08-1.03 (24H, m), 0.85 (3H, s), 0.56
(3H, s).
Reference Example 64
(3S,5S,10R,13R,14R)-3-Methoxymethoxy-10,13-dimethyl-17-triisopropylsilylox-
y
propan-2-yl-2,3,4,5,9,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]p-
henanthren-6(10H)-one (Compound P83)
[0376] Compound P82 (2.63 g, 4.79 mmol) obtained in Reference
Example 63 was treated with N-methylmorpholine oxide (3.36 g, 18.7
mmol) and tetrapropylammonium perruthenate (84.0 mg, 0.239 mmol) in
the same manner as Reference Example 4 to obtain the title compound
(2.50 g, 93%).
[0377] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.73 (1H, m),
4.72-4.67 (2H, m), 3.67 (1H, dd, J=3.1 Hz, 9.4 Hz), 3.53 (1H, m),
3.43 (1H, dd, J=6.4 Hz, 9.4 Hz), 3.38 (3H, s) 2.28 (1H, m),
2.20-1.23 (18H, m), 1.07-1.05 (24H, m), 0.87 (3H, s), 0.62 (3H,
s).
Reference Example 65
(1S,2R,5S)-5-Methoxymethoxy-2-methyl-2-{(3aR,7aR)-7a-methyl-4-oxo-1-[(S)-1-
-triisopropylsilyloxy
propan-2-yl]octahydro-1H-inden-5-yl}cyclohexanecarboxylic acid
(Compound P84)
[0378] Compound P83 (812 mg, 1.485 mmol) obtained in Reference
Example 64 was treated with sodium periodate (1.905 g, 8.908 mmol),
cerium chloride heptahydrate (553 mg, 1.49 mmol) and ruthenium
trichloride (92.0 mg, 0.446 mmol) in the same manner as Reference
Example 5, followed by treating the resulting crude product with
lead tetraacetate (984 mg, 2.00 mmol) to obtain the title compound
(296 mg, 35% (2 steps)).
[0379] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s), 3.64
(1H, dd, J=2.8 Hz, 9.7 Hz), 3.47-3.36 (5H, m), 2.68 (1H, m),
2.57-1.35 (19H, m), 1.25 (3H, s), 1.14-0.97 (24H, m), 0.60 (3H,
s).
Reference Example 66
(3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-1-[(S)-1-triisoprop-
yl silyloxy
propan-2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[-
h]chromen-5(10H)-one (Compound P85)
[0380] Compound P84 (295.7 mg, 0.5216 mmol) obtained in Reference
Example 65 was dissolved in dichloromethane (5 mL), and
triethylamine (0.73 mL, 5.22 mmol) and thionyl chloride (76 .mu.L,
1.04 mmol) were added thereto at 0.degree. C., followed by stirring
for 10 minutes. Water was added to the reaction mixture, followed
by extraction with chloroform (10 mL.times.3). The organic layer
was washed with saturated brine, and dried over anhydrous magnesium
sulfate. The residue obtained by concentration was purified by
silica gel column chromatography (25% ethyl acetate/n-hexane) to
obtain the title compound (125 mg, 47%).
[0381] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.71-4.68 (2H, m),
3.67 (1H, dd, J=3.3 Hz, 9.5 Hz), 3.52 (1H, m), 3.41-3.36 (4H, m),
2.36-1.23 (18H, m), 1.14-0.97 (24H, m), 0.97 (3H, s), 0.69 (3H,
s).
Reference Example 67
(10R,13R,14R)-3-Methoxymethoxy-10,13-dimethyl-17-[(S)-1-(2-methylallyloxyl-
)propan-2-yl]-2,3,4,9,10,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]-
phenanthrene (Compound P64)
[0382] Compound P57 (4.41 g, 7.49 mmol) obtained in Reference
Example 56 was treated with lithium aluminium hydride (1.42 g, 37.5
mmol) in the same manner as Reference Example 13 to obtain the
title compound (1.73 g, 54%).
[0383] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.57 (1H, m), 5.38
(1H, m), 4.95 (1H, m), 4.87 (1H, m), 4.71 (2H, s), 3.88-3.83 (2H,
m), 3.53 (1H, m), 3.37 (1H, m), 3.38 (3H, s), 3.13 (1H, dd, J=7.5
Hz, 9.0 Hz), 2.52 (1H, m), 2.36-2.32 (2H, m), 2.10-1.19 (18H, m),
1.08 (3H, d, J=6.6 Hz), 0.94 (3H, s), 0.63 (3H, s).
Reference Example 68
(5S,6S,10R,13R,14R)-17-[(2S)-1-(3-Hydroxy-2-methylpropoxy)propan-2-yl]-3-m-
ethoxymethoxy-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradeca-
hydro-1H-cyclopenta[a]phenanthren-6-ol (Compound P65)
[0384] Compound P64 (1.73 g, 4.03 mmol) obtained in Reference
Example 67 was treated with 1.0 mol/L of a borane-THF complex (12
mL, 12 mmol) in the same manner as Reference Example 2 to obtain
the title compound (960 mg, 51%).
[0385] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.18 (1H, s),
4.73-4.67 (2H, m), 3.80 (1H, m), 3.60-3.19 (8H, m), 3.38 (3H, s),
2.86 (1H, m), 2.31 (1H, m), 2.08-2.03 (2H, m), 1.89-1.13 (17H, m),
1.03 (3H, d, J=6.6 Hz), 0.87-0.84 (3H, m), 0.84 (3H, s), 0.55 (3H,
s).
Reference Example 69
(5S,10R,13R,14R)-17-[(2S)-1-(3-Hydroxy-2-methylpropoxy)propan-2-yl]-3-meth-
oxymethoxy-10,13-dimethyl-2,3,4,5,9,11,12,13,14,15,16,17-dodecahydro-1H-cy-
clopenta[a]phenanthren-6(10H)-one (Compound P66)
[0386] Compound P65 (960 mg, 7.06 mmol) obtained in Reference
Example 68 was dissolved in chloroform (80 mL), and manganese
dioxide (17.9 g, 206 mmol) was added, followed by stirring at room
temperature for 12 hours, and further stirring at 70.degree. C. for
1 hour. The reaction mixture was filtered through a celite to
obtain the title compound (955 mg, 100%).
[0387] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.72 (1H, m),
4.74-4.65 (2H, m), 3.64-3.18 (7H, m), 3.38 (3H, s), 2.30-1.21 (16H,
m), 1.05 (3H, d, J=6.6 Hz), 0.90-0.85 (6H, m), 0.61 (3H, s).
Reference Example 70
(5S,10R,13R,14R)-3-Methoxymethoxy-10,13-dimethyl-17-[(2S)-1-(2-methyl-3-tr-
iisopropylsilyloxypropoxy)propan-2-yl]-2,3,4,5,9,11,12,13,14,15,16,17-dode-
cahydro-1H-cyclopenta[a]phenanthren-6(10H)-one (Compound P67)
[0388] Compound P66 (955 mg, 2.06 mmol) obtained in Reference
Example 69 was treated with triisopropylsilyl chloride (2.20 mL,
10.3 mmol) in the same manner as Reference Example 63 to obtain a
crude product of the title compound (2.94 g).
Reference Example 71
(5S,7S,8S,10R,13R,14R)-7,8-Dihydroxy-3-methoxymethoxy-10,13-dimethyl-17-[(-
2S)-1-(2-methyl-3-triisopropylsilyloxypropoxy)propan-2-yl]tetradecahydro-1-
H-cyclopenta[a]phenanthren-6(10H)-one (Compound P68)
[0389] A crude product (2.94 g) of compound P67 obtained in
Reference Example 70 was treated with ruthenium trichloride (126
mg, 0.620 mmol), sodium periodate (2.65 g, 12.4 mmol) and cerium
chloride heptahydrate (770 mg, 2.06 mmol) in the same manner as
Reference Example 5 to obtain the title compound (608 mg, 45% (2
steps)).
[0390] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.74-4.67 (2H, m),
4.27 (1H, br s), 3.68-3.07 (9H, m), 3.38 (3H, s), 2.47-1.21 (19H,
m), 1.09-1.01 (27H, m), 0.93 (3H, d, J=4.4 Hz), 0.85 (3H, s).
Reference Example 72
(1S,2R)-5-Methoxymethoxy-2-methyl-2-{(1R,3aR,7aR)-7a-methyl-1-[(2S)-1-(2-m-
ethyl-3-triisopropylsilyloxypropoxy)propan-2-yl]-4-oxooctahydro-1H-inden-5-
-yl}cyclcohexanecarboxylic acid (Compound P69)
[0391] Compound P68 (608 mg, 0.93 mmol) obtained in Reference
Example 71 was treated with lead tetraacetate (826 mg, 1.86 mmol)
in the same manner as Reference Example 39 to obtain the title
compound (370 mg, 60%).
[0392] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.67 (2H, s),
3.63-3.08 (7H, m), 3.36 (3H, s), 2.71-1.18 (21H, m), 1.25 (3H, s),
1.06-1.04 (24H, m), 0.92 (3H, d, J=6.6 Hz), 0.60 (3H, s).
Reference Example 73
(1R,3aR,5aS,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-1-[(2S)-1-(2-methyl-
-3-triisopropylsilyloxypropoxy)propan-2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a--
dodecahydrobenzo[c]cyclopenta[h]chromen-5(10H)-one (Compound
P70)
[0393] Compound P69 (23.7 mg, 0.0370 mmol) obtained in Reference
Example 72 was treated with thionyl chloride (10.0 .mu.L, 0.14
mmol) in the same manner as Reference Example 40 to obtain the
title compound (13.6 mg, 59%).
[0394] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.70 (2H, m),
3.63-3.11 (7H, m), 3.38 (3H, s), 2.37-1.25 (19H, m), 1.10-1.02
(24H, m), 0.97 (3H, s), 0.92 (3H, d, J=6.8 Hz), 0.69 (3H, s).
Reference Example 74
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-chol-6,22-diene-24-propionic acid (Compound P71)
[0395] In THF (50 mL), 4-(carboxybutyl)triphenylphosphonium bromide
(22.5 g, 50.7 mmol) was suspended, followed by addition of
potassium tert-butoxide (11.4 g, 101 mmol). After the solution
turned orange color, compound P12 (4.20 g, 7.90 mmol) obtained in
Reference Example 12 was added, followed by stirring at room
temperature for 30 minutes. The mixture was ice-cooled, and then
1.0 mol/L of an aqueous solution of hydrochloric acid was added,
followed by extraction with ethyl acetate (100 mL.times.3). The
organic layer was washed with saturated brine, and dried over
anhydrous magnesium sulfate. The residue obtained by concentration
was purified by silica gel column chromatography (33 to 50% ethyl
acetate/n-hexane) to obtain the title compound (3.84 g, 80%).
[0396] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 8.16-8.10 (2H, m),
7.74-7.68 (2H, m), 6.66 (1H, d, J=8.1 Hz), 6.27 (1H, d, J=8.3 Hz),
5.22-5.16 (2H, m), 4.72-4.68 (2H, m), 4.11-3.92 (2H, m), 3.55 (1H,
m), 3.35 (3H, s), 2.39-1.33 (21H, m), 1.03 (3H, s), 0.97 (3H, d,
J=6.6 Hz), 0.82 (3H, s).
Reference Example 75
3-(Methoxymethyloxy)-5,8-(1,2,3,4-tetrahydro-1,4-dioxophthalazin-2,3-diyl)-
-chol-6-ene-24-propionic acid (Compound P72)
[0397] Compound P71 (3.93 g, 6.37 mmol) obtained in Reference
Example 74 was dissolved in ethyl acetate (227 mL). To the
resulting solution, 10% palladium-carbon (390 mg) was added,
followed by stirring at room temperature for 15 hours under a
hydrogen atmosphere. The catalyst was separated by filtration, and
then the filtrate was concentrated to obtain the title compound
(3.7 g).
Reference Example 76
(7R)-7-[(3S,10R,13R,14R,17R)-3-Methoxymethoxy-10,13-dimethyl-2,3,4,9,10,11-
,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl]octan-1-o-
l (Compound P73)
[0398] Compound P72 (3.7 g) obtained in Reference Example 75 was
treated with lithium aluminium hydride (1.10 g, 29.3 mmol) in the
same manner as Reference Example 13 to obtain the title compound
(1.4 g, 54% (2 steps)).
[0399] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.56 (1H, m), 5.39
(1H, m), 4.74-4.68 (2H, m), 3.70-3.60 (2H, m), 3.54 (1H, m), 3.38
(3H, s), 2.54-2.32 (3H, m), 2.05-1.19 (26H, m), 0.95-0.93 (6H, m),
0.62 (3H, s).
Reference Example 77
(7R)-7-[(3S,10R,13R,14R,17R)-3-Methoxymethoxy-10,13-dimethyl-2,3,4,9,10,11-
,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl]octyl
acetate (Compound P74)
[0400] Compound P73 (800 mg, 1.80 mmol) obtained in Reference
Example 76 was dissolved in pyridine (5 mL) and acetic anhydride (4
mL), followed by stirring at room temperature for 3.5 hours. The
resulting mixture was concentrated to obtain a crude product (841
mg) of the title compound.
Reference Example 78
(7R)-7-[(3S,5S,6S,10R,13R,14R,17R)-6-Hydroxy-3-methoxymethoxy-10,13-dimeth-
yl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phen-
anthren-17-yl]octyl acetate (Compound P75)
[0401] Compound P74 (841 mg) obtained in Reference Example 77 was
treated with 1.0 mol/L of a borane-THF complex (8.6 mL, 8.6 mmol)
in the same manner as Reference Example 2 to obtain the title
compound (241 mg, 48% (2 steps)).
[0402] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.34 (0.2H, br s),
5.18 (0.8H, br s), 4.75-4.68 (2H, m), 4.05 (2H, t, J=6.6 Hz), 3.81
(1H, m), 3.50 (1H, m), 3.38 (3H, s), 2.40-1.08 (33H, m), 0.92 (3H,
d, J=6.2 Hz), 0.85 (3H, s), 0.54 (3H, s).
Reference Example 79
(7R)-7-[(3S,5S,10R,13R,14R,17R)-3-Methoxymethoxy-10,13-dimethyl-6-oxo-2,3,-
4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-
-17-yl]octyl acetate (Compound P76)
[0403] Compound P75 (240.7 mg, 0.4769 mmol) obtained in Reference
Example 78 was treated with N-methylmorpholine oxide (83.8 mg,
0.715 mmol) and catalytic amount of tetrapropylammonium
perruthenate in the same manner as Reference Example 4 to obtain
the title compound (186 mg, 78%).
[0404] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.73 (1H, m),
4.74-4.68 (2H, m), 4.06 (2H, t, J=6.6 Hz), 3.53 (1H, m), 3.39 (3H,
s), 2.31-1.14 (32H, m), 0.94 (3H, d, J=6.1 Hz), 0.87 (3H, s), 0.60
(3H, s).
Reference Example 80
(7R)-7-[(3S,5S,7S,8S,10R,13R,14R,17R)-7,8-Dihydroxy-3-methoxymethoxy-10,13-
-dimethyl-6-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl]octyl
acetate (Compound P77)
[0405] Compound P76 (32.9 mg, 0.0654 mmol) obtained in Reference
Example 79 was treated with cerium chloride heptahydrate (24.0 mg,
0.0654 mmol), sodium periodate (84.0 mg, 0.392 mmol) and catalytic
amount of ruthenium chloride in the same manner as Reference
Example 5 to obtain the title compound (28 mg, 75%).
[0406] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
4.26 (1H, br s), 4.04 (2H, t, J=6.6 Hz), 3.56 (1H, m), 3.47 (1H,
s), 3.37-3.32 (1H, m), 3.36 (3H, s), 2.52-2.40 (1H, m), 2.03 (3H,
s), 2.14-1.10 (27H, m), 0.92-0.89 (6H, m), 0.83 (3H, s).
Reference Example 81
2-[1-(8-Acetoxyoctan-2-yl)-7a-methyl-4-oxooctahydro-1H-inden-5-yl]-5-metho-
xymethoxy-2-methylcyclohexane carboxylic acid (Compound P78)
[0407] Compound P77 (28.8 mg, 0.0537 mmol) obtained in Reference
Example 80 was treated with lead tetraacetate (48.0 mg, 0.107 mmol)
in the same manner as Reference Example 39 to obtain the title
compound (17.8 mg, 60%).
[0408] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73 (2H, s), 4.05
(2H, t, J=6.6 Hz), 3.45 (1H, m), 3.37 (3H, s), 2.67-0.83 (39H, m),
0.59 (3H, s).
Reference Example 82
(1S,2R,5S)-2-{(1R,3
aR,7aR)-1-[(R)-8-Hydroxyoctan-2-yl]-7a-methyl-4-oxooctahydro-1H-inden-5-y-
l}-5-methoxymethoxy-2-methylcyclohexanecarboxylic acid (Compound
P79)
[0409] Compound P78 (35.7 mg, 0.079 mmol) obtained in Reference
Example 81 was dissolved in methanol (3 mL), and then catalytic
amount of potassium carbonate was added thereto, followed by
stirring at room temperature for 5 hours. Water was added to the
reaction mixture, followed by extraction with ethyl acetate (10
mL.times.3). The organic layer was dried over anhydrous magnesium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (50% ethyl
acetate/n-hexane) to obtain the title compound (19.4 mg, 51%).
[0410] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.71 (2H, s), 3.64
(2H, t, J=6.6 Hz), 3.45 (1H, m), 3.36 (3H, s), 2.67-0.81 (37H, m),
0.59 (3H, s).
Reference Example 83
(1S,2R,5S)-2-{(1R,3aR,7aR)-1-[(R)-8-tert-Butyldimethylsiloxyoctan-2-yl]-7a-
-methyl-4-oxooctahydro-1H-inden-5-yl}-5-methoxymethoxy-2-methyleyclohexane-
carboxylic acid (Compound P80)
[0411] Compound P79 (19.4 mg, 0.0400 mmol) obtained in Reference
Example 82 was dissolved in DMF (3 mL), and imidazole (54.0 mg,
0.80 mmol) and tert-butyldimethylsilyl chloride (60.0 mg, 0.40
mmol) were added thereto, followed by stirring at room temperature
for 3.5 hours. Water was added to the reaction mixture, followed by
extraction with diethyl ether (10 mL.times.3). The organic layer
was washed with saturated brine, dried over anhydrous magnesium
sulfate, and concentrated to yield a residue. The residue was
purified by silica gel column chromatography (33% ethyl
acetate/n-hexane) to obtain the title compound (11.4 mg, 48%).
[0412] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.62 (2H, s), 3.55
(2H, t, J=6.6 Hz), 3.32 (3H, s), 2.60-1.05 (31H, m), 0.86-0.85
(21H, m), 0.54 (3H, s).
Reference Example 84
(1R,3aR,5aR,7S,9aS)-1-[(R)-8-tert-Butyldimethylsiloxyoctan-2-yl-7-methoxym-
ethoxy]-9a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]c-
yclopenta[h]chromen-5(10H)-one (Compound P81)
[0413] Compound P80 (11.4 mg; 0.0190 mmol) obtained in Reference
Example 83 was treated with thionyl chloride (7.0 .mu.L, 0.095
mmol) in the same manner as Reference Example 40 to obtain the
title compound (0.9 mg, 8%).
[0414] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.62-3.50 (3H, m), 3.38 (3H, s), 2.37-1.20 (28H, m), 0.94-0.90
(15H, m), 0.67 (3H, s), 0.05 (6H, s).
Reference Example 85
7-Oxa-ergosta-8(9),24(28)-dien-6-on-3-ol
[0415] As the first seed medium and second seed medium, a medium
comprising glucose (20 g/L), mashed potatoes (30 g/L) and dry yeast
extract (5 g/L) (pH 6.5) was used, and as the main fermentation
medium, a medium comprising sucrose (30 g/L), soluble starch (20
g/L), corn steep liquor (CSL) (30 g/L) and calcium carbonate (5
g/L) (pH 5.0) was used. A piece of agar containing Penicillium sp.
CND1007 (FERM BP-10917) was inoculated into the first seed medium
(10 mL) which had been added into a 70 mL capacity test tube,
followed by shaking culturing at 28.degree. C. for 72 hours. Next,
25 mL per flask of the first seed culture liquid was inoculated
into the second seed medium (475 mL) which had been added into each
of 2 L capacity conical flasks, followed by shaking for 72 hours in
the same manner. Subsequently, 900 mL per fermenter of the second
seed culture liquid was inoculated into the main fermentation
medium (about 54 L) which had been dispensed into three 30 L
capacity jar fermenters, followed by agitation culturing (the
number of revolutions 250 rpm) at 25.degree. C. for 8 days.
Additionally, 25 mL per flask of the second seed culture liquid was
inoculated into the main fermentation medium (about 10 L) which had
been dispensed into 20 conical flasks each having 2 L capacity,
followed by agitation culturing (the number of revolutions 220 rpm)
at 25.degree. C. for 8 days.
[0416] A filter aid (Radiolite #600, manufactured by Showa Chemical
Industry) was added at a ratio of 10% by weight to the thus
obtained fermentation culture liquid (64 L) and then the culture
filtrate and cells were separated by suction filtration. The
separated cells were mixed with 15 L of methanol, followed by
extraction twice at room temperature. The extract (30 L) was
concentrated to 10 L under reduced pressure and applied to a column
filled with 2 L of Diaion HP 20 (manufactured by Mitsubishi
Chemical Corp.) to adsorb the desired compound. After washing with
water, 40% methanol and 70% methanol, the desired compound was
eluted with 100% methanol and 30% acetone/methanol. The eluate (6
L) was concentrated to 1 L under reduced pressure and then
extracted three times with chlorofolin (1 L). The residue (15 g)
obtained by concentrating the extract under reduced pressure was
applied to a column filled with 500 mL of silica gel and eluted
stepwise using n-hexane, ethyl acetate, methanol and a mixed
solvent thereof. Components contained in each eluate was detected
by thin layer chromatography, and eluates containing the same
component were combined to obtain fractions of 20 to 40% ethyl
acetate/n-hexane elution fraction (fraction 1), 60% ethyl
acetate/n-hexane elution fraction (fraction 2), 60 to 80% ethyl
acetate/n-hexane elution fraction (fraction 3), 80% ethyl
acetate/n-hexane to ethyl acetate elution fraction (fraction 4),
ethyl acetate to 25% methanol/ethyl acetate elution fraction
(fraction 5) and 25% methanol/ethyl acetate elution fraction
(fraction 6).
[0417] The fraction 2 (300 mg) was applied to a column filled with
20 mL of silica gel, followed by elution with n-hexane and ethyl
acetate. Fractions containing the compounds of interest were
collected and concentrated, and the thus obtained residue (200 mg)
was separated and purified by fractional high performance liquid
chromatography [column SunFire.TM. Prep C18 OBD 10 .mu.m,
.phi.19.times.250 mm, column temperature 40.degree. C., flow rate
10 mL/min, stepwise elution with 85 to 100% methanol aqueous
solution] to obtain 7-oxa-ergosta-8(9),24(28)-dien-6-on-3-ol (44.6
mg).
EXAMPLE 1
(S)-7-Oxa-8-cholesten-3,6-dione (Compound 1)
[0418] Compound P5 (167 mg, 0.399 mmol) obtained in Reference
Example 5 was dissolved in dichloromethane (4.0 mL), and
triethylamine (166 .mu.L, 1.20 mmol) and thionyl chloride (43.7
.mu.L, 0.598 mmol) were added thereto at room temperature, followed
by stirring for 20 minutes. A saturated aqueous sodium bicarbonate
solution was added to the reaction mixture, followed by extraction
with chloroform twice. The organic layer was washed with saturated
brine, dried over anhydrous magnesium sulfate, and concentrated
under reduced pressure to yield a residue. The residue was purified
by silica gel column chromatography to obtain the title compound
(133 mg, 83%).
[0419] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.71 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.94 (d, J=6.6 Hz, 3H),
0.99-2.63 (m, 26H), 2.70-2.82 (m, 2H).
EXAMPLE 2
(3S,5S)-3-Hydroxy-7-oxa-8-cholesten-6-one (Compound 2)
[0420] Compound 1 (400 mg, 0.999 mmol) obtained in Example 1 was
dissolved in a mixed solvent of methanol (6.0 mL) and
dichloromethane (4.0 mL), and sodium borohydride (37.8 mg, 0.999
mmol) was added thereto at 0.degree. C., followed by stirring at 20
minutes. A saturated aqueous ammonium chloride solution was added
to the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain the title compound (363 mg,
90%).
[0421] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H),
0.97 (s, 3H), 1.00-2.43 (m, 25H), 3.57-3.69 (m, 1H).
EXAMPLE 3
(S)-3-Hydroxyimino-7-oxa-8-cholesten-6-one (Compound 3)
[0422] Compound 1 (172 mg, 0.429 mmol) obtained in Example 1 was
dissolved in ethanol (4.0 mL), and hydroxyamine hydrochloride (89.5
mg, 1.29 mmol) and triethylamine (178 .mu.L, 1.29 mmol) were added
thereto at room temperature, followed by stirring for 20 minutes. A
saturated aqueous ammonium chloride solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was washed with saturated brine, dried over
anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain the title compound (160 mg, 90%) as
a mixture of E-form and Z-form.
[0423] ESI-MS: m/z 416[M+H].sup.+, NMR (CDCl.sub.3) .delta.(ppm):
0.69 (s, 3H), 0.86 (d, J=6.4 Hz, 3H), 0.88 (d, J=6.4 Hz, 3H), 0.93
(d, J=6.3 Hz, 3H), 0.98-2.80 (m, 27H), 3.30-3.40 (m, 0.5H),
3.68-3.78 (m, 0.5H), 6.77 (s, 0.5H), 6.81 (s, 0.5H).
EXAMPLE 4
(S)-7-Oxa-1,8-cholestadien-3,6-dione (Compound 4) and
7-oxa-4,8-cholestadien-3,6-dione (Compound 5)
[0424] Compound 1 (207 mg, 0.516 mmol) obtained in Example 1 was
dissolved in THF (5.0 mL), and chlorotrimethylsilane (197 .mu.L,
1.55 mmol) and 1.0 mol/L of a lithium bis(trimethylsilylamide)/THF
solution (723 .mu.L, 0.723 mmol) were added thereto at 0.degree.
C., followed by stirring for 1 hour. A saturated aqueous ammonium
chloride solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure. The resulting residue was
suspended in acetonitrile (20 mL), and then palladium acetate (232
mg, 1.03 mmol) was added thereto, followed by stirring at room
temperature for 3.5 hours. The reaction mixture was filtrated
through a celite, and the filtrate was concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain compound 4 (98.2 mg, 48%) and
compound 5 (47.3 mg, 23%).
[0425] Compound 4; ESI-MS: m/z 399 [M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.72 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.95 (d, J=6.6 Hz, 3H), 1.00-1.83 (m, 16H),
1.94-2.05 (m, 1H), 2.10-2.19 (m, 1H), 2.31-2.43 (m, 3H), 2.66 (dd,
J=13.8, 18.0 Hz, 1H), 2.89 (dd, J=4.4, 18.0 Hz, 1H), 3.16 (dd,
J=4.4, 13.8 Hz, 1H), 6.00 (d, J=9.9 Hz, 1H), 7.08 (d, J=9.9 Hz,
1H).
[0426] Compound 5; ESI-MS: m/z 399[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.74 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.98-1.60 (m, 15H),
1.76-1.87 (m, 1H), 1.95-2.13 (m, 4H), 2.17-2.26 (m, 2H), 2.32-2.40
(m, 1H), 2.52-2.61 (m, 2H), 6.72 (s, 1H).
EXAMPLE 5
(1R,2R,5S)-1,2-Epoxy-7-oxa-8-cholesten-3,6-dione (Compound 6)
[0427] Compound 4 (11.0 mg, 0.0276 mmol) obtained in Example 4 was
dissolved in THF (1.0 mL), and 30% aqueous hydrogen peroxide (8.5
.mu.L, 0.0828 mmol) and 10% aqueous sodium hydroxide solution (20.1
.mu.L, 0.0551 mmol) were added thereto at 0.degree. C., followed by
stirring at room temperature for 40 minutes. A saturated aqueous
ammonium chloride solution and a saturated aqueous sodium
thiosulfate solution were added to the reaction mixture, followed
by extraction with ethyl acetate twice. The organic layer was
washed with saturated brine, dried over anhydrous magnesium
sulfate, and concentrated under reduced pressure to yield a
residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (4.0 mg, 35%).
[0428] ESI-MS: m/z 415[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.72 (s, 3H), 0.86 (d, J=6.7 Hz, 3H), 0.88 (d, J=6.7
Hz, 3H), 0.96 (d, J=6.6 Hz, 3H), 0.98-1.60 (m, 15H), 1.72-1.82 (m,
1H), 1.94-2.05 (m, 1H), 2.10-2.19 (m, 1H), 2.28-2.57 (m, 4H), 2.74
(dd, J=6.3, 19.8 Hz, 1H), 3.25 (dd, J=6.3, 12.6 Hz, 1H), 3.36 (d,
J=4.3 Hz, 1H), 3.62 (d, J=4.3 Hz, 1H).
EXAMPLE 6
(5)-3-Methylen-7-oxa-8-cholesten-6-one (Compound 7) and
(R)-3-methylen-7-oxa-8-cholesten-6-one (Compound 9)
[0429] Methyl triphenylphosphonium bromide (58.0 mg, 0.162 mmol)
was suspended in THF (1.0 mL), followed by adding 1.0 mol/L of a
lithium bis(trimethylsilylamide)/THF solution (225 .mu.L, 0.225
mmol) at 0.degree. C. Subsequently, a THF solution (0.5 mL) of
compound 1 (50.0 mg, 0.125 mmol) obtained in Example 1 was added
thereto, followed by stirring at room temperature for 1 hour. Water
was added to the reaction mixture, followed by extraction with
ethyl acetate twice. The organic layer was washed with saturated
brine, dried over anhydrous sodium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 7 (10.3
mg, 21%) and compound 9 (13.1 mg, 26%).
[0430] Compound 7; ESI-MS: m/z 399[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.97-1.54 (m, 16H),
1.69-1.78 (m, 2H), 1.90-2.44 (m, 9H), 2.59-2.65 (m, 1H), 4.71-4.76
(m, 2H).
[0431] Compound 9; ESI-MS: m/z 399[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.70 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.98-1.57 (m, 17H),
1.70-2.27 (m, 8H), 2.31-2.45 (m, 3H), 4.71-4.76 (m, 2H).
EXAMPLE 7
(Z)-3-Hydroxyimino-7-oxa-4,8-cholestadien-6-one (Compound 8)
[0432] The title compound (14.4 mg, 51%) was obtained by using
Compound 5 (27.4 mg, 0.0687 mmol) obtained in Example 4 in the same
manner as Example 3.
[0433] ESI-MS: m/z 414[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.73 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98-2.10 (m, 21H), 2.17-2.38 (m,
4H), 3.06-3.14 (m, 1H), 7.21 (s, 1H).
EXAMPLE 8
(3R,5S)-3-Ethylamino-7-oxa-8-cholesten-6-one (Compound 10) and
(3S,5S)-3-ethylamino-7-oxa-8-cholesten-6-one (Compound 11)
[0434] Compound 1 (19.7 mg, 0.0492 mmol) obtained in Example 1 was
dissolved in THF (5.0 mL), and 2.0 mol/L of an ethylamine/THF
solution (1.0 mL, 2.00 mmol) and powered Molecular Sieves 4 .ANG.
(1.0 g) were added thereto at room temperature, followed by
stirring overnight. Subsequently, sodium triacetoxyborohydride (208
mg, 0.492 mmol) was added to the reaction mixture, followed by
stirring for 30 minutes. A saturated aqueous ammonium chloride
solution was added to the reaction mixture and filtered through a
cotton plug, followed by extraction with chloroform five times. The
organic layer was dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain
compound 10 (4.1 mg, 19%) and compound 11 (4.8 mg, 23%).
[0435] Compound 10; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s,
3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.92 (d, J=6.6
Hz, 3H), 0.95 (s, 3H), 0.99-2.37 (m, 27H), 2.53-2.65 (m, 2H), 2.93
(dd, J=3.3, 12.9 Hz, 1H), 3.00-3.06 (m, 1H).
[0436] Compound 11; ESI-MS: m/z 430[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=7.0 Hz, 3H),
0.88 (d, J=7.0 Hz, 3H), 0.92 (d, J=6.6 Hz, 3H), 0.95 (s, 3H),
0.99-1.83 (m, 21H), 1.90-2.56 (m, 8H), 2.60-2.80 (m, 2H).
EXAMPLE 9
(R)-7-Oxa-1,8-cholesten-3,6-dione (Compound 12)
[0437] Compound 4 (30.0 mg, 0.0753 mmol) obtained in Example 4 was
dissolved in THF (2.0 mL), and 1,8-diazabicyclo[5.4.0]undeca-7-ene
(80.0 .mu.L, 0.534 mmol) was added thereto at room temperature,
followed by stirring for 2 hours. A saturated aqueous ammonium
chloride solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain the
title compound (22.3 mg, 74%).
[0438] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.76 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.94 (d, J=6.6 Hz, 3H),
0.99-1.85 (m, 16H), 1.95-2.21 (m, 3H), 2.26-2.43 (m, 2H), 2.63 (dd,
J=4.0, 16.5 Hz, 1H), 2.92 (dd, J=8.8, 16.5 Hz, 1H), 3.01 (dd,
J=4.0, 8.8 Hz, 1H), 5.98 (d, J=9.9 Hz, 1H), 6.65 (d, J=9.9 Hz,
1H).
EXAMPLE 10
(3S,5S)-3-Amino-7-oxa-8-cholesten-6-one (Compound 13) and
(3R,5S)-3-hydroxy-7-oxa-8-cholesten-6-one (Compound 105)
[0439] Step 1; Compound 1 (359 mg, 0.896 mmol) obtained in Example
1 was dissolved in THF (10 mL), and 1.0 mol/L of a potassium
tri(sec-butyl)borohydride/THF solution (1.61 mL, 1.61 mmol) was
added thereto at -78.degree. C., followed by stirring for 50
minutes. A saturated aqueous ammonium chloride solution was added
to the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain compound 105 (157 mg, 39%).
[0440] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H),
0.94 (s, 3H), 0.99-2.42 (m, 24H), 2.91 (dd, J=3.6, 12.6 Hz, 1H),
4.22 (m, 1H).
[0441] Step 2; (3R,5S)-3-Hydroxy-7-oxa-8-cholesten-6-one (70.0 mg,
0.174 mmol) obtained in the Step 1 and triphenylphosphine (73.0 mg,
0.278 mmol) were dissolved in toluene (1.5 mL), and 2.2 mol/L of an
diethyl azodicarboxylate/toluene solution (119 .mu.L, 0.261 mmol)
and diphenyl phosphorazidate (112 .mu.L, 0.522 mmol) were added
thereto at 0.degree. C., followed by stirring for 1 hour.
Subsequently, 2.2 mol/L of an diethyl azodicarboxylate/toluene
solution (39.7 .mu.L, 0.0869 mmol) and triphenylphosphine (22.8 mg,
0.0869 mmol) were further added, followed by stirring for 20
minutes, and then stirring at room temperature for 1 hour. The
residue obtained by concentration under reduced pressure was
purified by silica gel column chromatography to obtain
(3S,5S)-3-azido-7-oxa-8-cholesten-6-one (33.5 mg, 45%).
[0442] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.76 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H),
0.97 (s, 3H), 1.00-1.79 (m, 17H), 1.88-2.43 (m, 8H), 3.25-3.36 (m,
1H).
[0443] Step 3; (3S,5S)-3-Azido-7-oxa-8-cholesten-6-one (33.5 mg,
0.0783 mmol) obtained in the Step 2 was dissolved in a mixed
solvent of THF (1.0 mL) and water (0.1 mL), and triphenylphosphine
(41.0 mg, 0.157 mmol) was added thereto at room temperature,
followed by stirring overnight. The residue obtained by
concentration under reduced pressure was purified by silica gel
column chromatography to obtain compound 13 (27.3 mg, 87%).
[0444] ESI-MS: m/z 402[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.95 (s, 3H), 0.97-1.60 (m, 15H),
1.63-1.82 (m, 3H), 1.90-2.43 (m, 7H), 2.65-2.75 (m, 1H).
EXAMPLE 11
(3S,5S)-3-N-Methanesulfonylamino-7-oxa-8-cholesten-6-one (Compound
14)
[0445] Compound 13 (7.0 mg, 0.017 mmol) obtained in the Step 3 of
Example 10 was dissolved in dichloromethane (0.5 mL), and
triethylamine (3.6 .mu.L, 0.026 mmol) and methanesulfonyl chloride
(2.0 .mu.L, 0.026 mmol) were added thereto at room temperature,
followed by stirring for 20 minutes. A saturated aqueous ammonium
chloride solution was added to the reaction mixture, followed by
extraction with chloroform twice. The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain the
title compound (6.2 mg, 74%).
[0446] ESI-MS: m/z 480[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.95 (s, 3H), 0.98-1.75 (m, 17H),
1.92-2.47 (m, 8H), 3.01 (s, 3H), 3.27-3.38 (m, 1H), 4.29-4.33 (m,
1H).
EXAMPLE 12
(3S,5S)-3-N-Phenylcarbamoylamino-7-oxa-8-cholesten-6-one (Compound
15)
[0447] The title compound (5.0 mg, 67%) was obtained by using
Compound 13 (5.8 mg, 0.014 mmol) obtained in the Step 3 of Example
10 and phenyl isocyanate (2.4 .mu.L, 0.022 mmol) in the same manner
as Example 11.
[0448] ESI-MS: m/z 521[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.91 (s, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.98-1.79 (m, 17H),
1.90-2.45 (m, 8H), 3.63-3.73 (m, 1H), 4.93-4.98 (m, 1H), 6.68 (s,
1H), 7.01-7.11 (m, 1H), 7.28-7.31 (m, 4H).
EXAMPLE 13
(3S,5S)-3-N-Benzoylamino-7-oxa-8-cholesten-6-one (Compound 16)
[0449] The title compound (7.1 mg, 87%) was obtained by using
Compound 13 (6.5 mg, 0.016 mmol) obtained in the Step 3 of Example
10 and benzoyl chloride (2.8 .mu.L, 0.024 mmol) in the same manner
as Example 11.
[0450] ESI-MS: m/z 506[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.98 (s, 3H), 1.00-1.78 (m, 17H),
1.92-2.56 (m, 8H), 3.96-4.07 (m, 1H), 6.03-6.09 (m, 1H), 7.40-7.53
(m, 3H), 7.71-7.78 (m, 2H).
EXAMPLE 14
(3S,5S)-3-N-Acetylamino-7-oxa-8-cholesten-6-one (Compound 17)
[0451] Compound 13 (4.2 mg, 0.0105 mmol) obtained in the Step 3 of
Example 10 was dissolved in a mixed solvent of acetic anhydride
(0.5 mL) and pyridine (0.25 mL), followed by stirring for 50
minutes. The residue obtained by concentration under reduced
pressure was purified by preparative thin layer chromatography to
obtain the title compound (4.0 mg, 86%).
[0452] ESI-MS: m/z 444[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.95 (s, 3H), 0.99-1.78 (m, 17H),
1.90-2.49 (m, 11H), 3.75-3.84 (m, 1H), 5.34-5.39 (m, 1H).
EXAMPLE 15
(1R,2R,5S)-1,2-Dihydroxy-7-oxa-8-cholesten-3,6-dione (Compound
18)
[0453] Compound 4 (61.7 mg, 0.155 mmol) obtained in Example 4 was
dissolved in a mixed solvent of tert-butyl alcohol (4.8 mL) and
water (0.8 mL), and 1% of an aqueous solution of potassium
permanganate (2.39 mL, 0.155 mmol) was added thereof at 0.degree.
C., followed by stirring at 15 minutes. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
filtering through a celite, and the filtrate was extracted with
ethyl acetate twice. The organic layer was washed with saturated
brine, dried over anhydrous magnesium sulfate, and concentrated
under reduced pressure to yield a residue. The residue was purified
by preparative thin layer chromatography to obtain the title
compound (22.5 mg, 34%).
[0454] ESI-MS: m/z 433[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.71 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.97-1.79 (m, 15H), 1.92-2.11 (m,
2H), 2.17-2.30 (m, 1H), 2.35-2.46 (m, 1H), 2.50-2.79 (m, 3H), 2.95
(dd, 3=4.4, 15.8 Hz, 1H), 3.41 (dd, J=4.4, 13.2 Hz, 1H), 3.75-3.85
(br s, 1H), 4.19 (d, J=2.9 Hz, 1H), 4.28 (d, J=2.9 Hz, 1H).
EXAMPLE 16
(3S,5S)-3-Hydroxy-7-oxa-1,8-cholestadien-6-one (Compound 19)
[0455] Compound 4 (19 mg, 0.048 mmol) obtained in Example 4 was
suspended in methanol (1.0 mL), and cerium chloride heptahydrate
(36 mg, 0.096 mmol) and sodium borohydride (0.9 mg, 0.024 mmol)
were added thereof at 0.degree. C., followed by stirring at room
temperature for 10 minutes. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain the
title compound (17 mg, 91%).
[0456] ESI-MS: m/z 401[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.69 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.97-2.12 (m, 19H), 2.25-2.36 (m,
3H), 2.52-2.60 (m, 1H), 2.69 (dd, J=2.4, 13.2 Hz, 1H), 4.32-4.41
(m, 1H), 5.62-5.69 (m, 1H), 5.83 (dd, J=2.0, 10.2 Hz, 1H).
EXAMPLE 17
(3S,5R)-3-Hydroxy-5-methyl-7-oxa-8-cholesten-6-one (Compound 20),
(3S,5R)-3-methoxy-5-methyl-7-oxa-8-cholesten-6-one (Compound 22),
and (3S,5S)-3-methoxy-5-methyl-7-oxa-8-cholesten-6-one (Compound
23)
[0457] Compound 2 (67 mg, 0.17 mmol) obtained in Example 2 was
dissolved in DMF (2.0 mL), and silver oxide (120 mg, 0.50 mmol),
iodomethane (52 .mu.L, 0.84 mmol) and potassium iodide (83 mg, 0.50
mmol) were added under light shading at room temperature, followed
by stirring for 1 hour. Ethyl acetate was added until the reaction
mixture became white turbidity, followed by filtration through a
celite. The filtrate was washed with water and saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 20 (3.6
mg, 5%), compound 22 (44 mg, 61%), and compound 23 (20 mg,
28%).
[0458] Compound 20; ESI-MS: 417[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.78 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.98-1.85 (m, 24H),
1.92-2.33 (m, 5H), 2.41-2.48 (m, 1H), 3.82-3.93 (m, 1H).
[0459] Compound 22; ESI-MS: m/z 431[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.77 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98-2.34 (m, 29H),
2.50-2.57 (m, 1H), 3.32-3.45 (m, 1H), 3.38 (s, 3H).
[0460] Compound 23; ESI-MS: m/z 431[M+H].sup.+, NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.98-1.82 (m, 23H), 1.90-2.28 (m,
5H), 2.31-2.48 (m, 2H), 3.35-3.46 (m, 1H), 3.36 (s, 3H).
EXAMPLE 18
(1S,5S)-1-Hydroxy-7-oxa-2,8-cholestadien-6-one (Compound 21)
[0461] Compound 6 (9.0 mg, 0.022 mmol) obtained in Example 5 was
dissolved in ethanol (0.5 mL), and hydrazine monohydrate (10 .mu.L)
and acetic acid (10 .mu.L) were added thereto at room temperature,
followed by stirring for 1 hour. Water was added to the reaction
mixture, followed by extraction with ethyl acetate twice. The
organic layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (6.2 mg, 71%).
[0462] ESI-MS: 401[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.69 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.90 (s, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.97-1.81 (m, 13H),
1.91-2.09 (m, 2H), 2.17-2.59 (m, 5H), 3.02 (dd, J=6.0, 11.4 Hz,
1H), 3.92-3.98 (m, 1H), 5.84-5.98 (m, 2H).
EXAMPLE 19
(3S,5R)-3-Hydroxy-5-allyl-7-oxa-8-cholesten-6-one (Compound 24) and
(3S,5S)-3-hydroxy-5-allyl-7-oxa-8-cholesten-6-one (Compound 30)
[0463] Step 1; Compound 2 (109 mg, 0.270 mmol) obtained in Example
2 was dissolved in dichloromethane (1.0 mL), and imidazole (36.8
mg, 0.541 mmol) and chlorotriethylsilane (68.1 .mu.L, 0.406 mmol)
were added thereto at room temperature, followed by stirring for 20
minutes. A saturated aqueous ammonium chloride solution was added
to the reaction mixture, followed by extraction with chloroform
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain
(3S,5S)-3-triethylsiloxy-7-oxa-8-cholesten-6-one (124 mg, 89%).
[0464] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.59 (q, J=7.9 Hz,
6H), 0.67 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H),
0.91-0.99 (m, 15H), 1.01-1.79 (m, 19H), 1.90-2.40 (m, 6H),
3.51-3.63 (m, 1H).
[0465] Step 2; By using
(3S,5S)-3-triethylsiloxy-7-oxa-8-cholesten-6-one (42.0 mg, 0.0812
mmol) obtained in the Step 1, silver oxide (56.4 mg, 0.243 mmol),
allyl bromide (35.2 .mu.L, 0.406 mmol) and potassium iodide (40.0
mg, 0.243 mmol),
(3S,5R)-3-tiethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one (22.1 mg,
49%) and (3S,5S)-3-triethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one
(11.0 mg, 24%) were obtained in the same manner as Example 17.
[0466] (3S,5R)-3-Tiethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one;
[0467] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.62 (q, J=7.8 Hz,
6H), 0.79 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H),
0.93 (d, J=6.3 Hz, 3H), 0.96 (t, J=7.8 Hz, 9H), 1.01-2.32 (m, 27H),
2.41-2.51 (m, 2H), 3.75-3.86 (m, 1H), 4.95-5.01 (m, 2H), 5.59-5.74
(m, 1H).
[0468]
(3S,5S)-3-Triethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one;
[0469] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.57 (q, J=7.8 Hz,
6H), 0.68 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H),
0.91-1.00 (m, 12H), 1.06-1.81 (m, 21H), 1.91-2.53 (m, 8H),
3.74-3.84 (m, 1H), 5.04-5.14 (m, 2H), 5.58-5.72 (m, 1H).
[0470] Step 3;
(3S,5R)-3-Triethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one (4.0 mg,
0.0072 mmol) obtained in the Step 2 was dissolved in a mixed
solvent of ethanol (0.8 mL) and water (0.1 mL), and sodium
periodate (4.6 mg, 0.022 mmol) was added thereto at room
temperature, followed by stirring for 30 minutes. A saturated
aqueous sodium thiosulfate solution was added to the reaction
mixture, followed by extraction with ethyl acetate twice. The
organic layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by preparative thin layer
chromatography to obtain compound 24 (1.8 mg, 57%).
[0471] ESI-MS: m/z 443[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.80 (s, 3H), 0.86 (d, J=6.8 Hz, 3H), 0.88 (d, J=6.8
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 1.01-2.35 (m, 27H), 2.46-2.57 (m,
2H), 3.82-3.96 (m, 1H), 4.98-5.12 (m, 2H), 5.59-5.72 (m, 1H).
[0472] Step 4;
(3S,5S)-3-Triethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one (5.0 mg,
0.0090 mmol) obtained in the Step 2 was dissolved in THF (1.0 mL),
and acetic acid (1.0 .mu.L, 0.018 mmol) and 1.0 mol/L of a
tetrabutylammonium fluoride/THF solution (18 .mu.L, 0.018 mmol)
were added thereto at room temperature, followed by stirring for 2
hours. A saturated aqueous ammonium chloride solution was added to
the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 30 (3.1
mg, 78%).
[0473] ESI-MS: m/z 443[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.69 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.99-2.53 (m, 29H), 3.80-3.92 (m,
1H), 5.04-5.14 (m, 2H), 5.59-5.73 (m, 1H).
EXAMPLE 20
(3S,5R)-3-Hydroxy-5-ethyl-7-oxa-8-cholesten-6-one (Compound 25)
[0474] Step 1; By using
(3S,5S)-3-triethylsiloxy-7-oxa-8-cholesten-6-one (9.0 mg, 0.017
mmol) obtained in the Step 1 of Example 19, silver oxide (8.0 mg,
0.035 mmol), iodoethane (14 .mu.L, 0.174 mmol) and potassium iodide
(5.8 mg, 0.035 mmol),
(3S,5R)-3-triethylsiloxy-5-ethyl-7-oxa-8-cholesten-6-one (4.9 mg,
52%) was obtained in the same manner as Example 17.
[0475] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.63 (q, J=8.1 Hz,
6H), 0.76 (s, 3H), 0.81 (t, J=7.7 Hz, 3H), 0.86 (d, J=6.9 Hz, 3H),
0.88 (d, J=6.9 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.97 (t, J=8.1 Hz,
9H), 1.01 (s, 3H), 1.05-1.84 (m, 20H), 1.91-2.32 (m, 5H), 2.45-2.53
(m, 1H), 3.75-3.87 (m, 1H).
[0476] Step 2; By using
(3S,5R)-3-triethylsiloxy-5-ethyl-7-oxa-8-cholesten-6-one (4.9 mg,
0.0090 mmol) obtained in the Step 1, compound 25 (3.0 mg, 77%) was
obtained in the same manner as the Step 4 of Example 19.
[0477] ESI-MS: m/z 431[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.77 (s, 3H), 0.83 (t, J=7.0 Hz, 3H), 0.86 (d, J=7.0
Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 1.01-2.32
(m, 28H), 2.55-2.63 (m, 1H), 3.83-3.97 (m, 1H).
EXAMPLE 21
(3S,5S)-3-Hydroxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
(Compound 26 and Compound 27)
[0478] Step 1;
(3S,5R)-3-Triethylsiloxy-5-allyl-7-oxa-8-cholesten-6-one (23 mg,
0.042 mmol) obtained in the Step 2 of Example 19 was dissolved in a
mixed solvent of tert-butyl alcohol (0.8 mL) and water (0.2 mL),
and 4-methylmorpholin-N-oxide (7.3 mg, 0.062 mmol) and 2.5% of an
osmium tetroxide/tert-butyl alcohol solution (26 .mu.L, 0.00207
mmol) were added thereto at room temperature, followed by stirring
overnight at room temperature. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to isolate
each of the stereoisomer of
(3S,5R)-3-triethylsiloxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
(Low polar compound; 8.3 mg, 34%, High polar compound; 12 mg,
48%).
[0479] Low polar compound; .sup.1H NMR (CDCl.sub.3) .delta.(ppm):
0.63 (q, J=7.8 Hz, 6H), 0.79 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88
(d, J=7.0 Hz, 3H), 0.91-1.00 (m, 15H), 1.04-1.82 (m, 18H),
1.89-2.35 (m, 7H), 2.47-2.57 (m, 1H), 3.36 (dd, J=8.1, 10.6 Hz,
1H), 3.49 (dd, J=4.0, 10.6 Hz, 1H), 3.76-3.98 (m, 2H).
[0480] High polar compound; .sup.1H NMR (CDCl.sub.3) .delta.(ppm):
0.63 (q, J=8.1 Hz, 6H), 0.73 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88
(d, J=7.0 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.97 (t, J=8.1 Hz, 9H),
0.99-2.34 (m, 28H), 2.65-2.78 (m, 1H), 3.33 (dd, J=7.3, 11.0 Hz,
1H), 3.44-3.52 (m, 1H), 3.62-3.72 (m, 1H), 3.74-3.86 (m, 1H).
[0481] Step 2; By using the stereoisomer (Low polar compound; 5.5
mg, 0.0093 mmol) of
(3S,5R)-3-triethylsiloxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
obtained in the Step 1,
(3S,5S)-3-hydroxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
(Compound 26 or Compound 27) (3.6 mg, 81%) was obtained in the same
manner as the Step 4 of Example 19.
[0482] ESI-MS: m/z 477[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.80 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.98-2.35 (m, 28H), 2.58-2.68 (m,
1H), 3.38 (dd, J=7.6, 10.9 Hz, 1H), 3.50 (dd, J=4.0, 10.9 Hz, 1H),
3.84-3.98 (m, 2H).
[0483] Step 3; By using the stereoisomer (High polar compound; 6.0
mg, 0.010 mmol) of
(3S,5R)-3-triethylsiloxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
obtained in the Step 1,
(3S,5S)-3-hydroxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-6-one
(Compound 27 or Compound 26) (3.7 mg, 76%) was obtained in the same
manner as the Step 4 of Example 19.
[0484] ESI-MS: m/z 477[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.73 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.98-2.34 (m, 28H), 2.45-2.55 (m,
1H), 3.28-4.00 (m, 4H).
EXAMPLE 22
(3S,5R)-3-Hydroxy-5-(2hydroxy)ethyl-7-oxa-8-cholesten-6-one
(Compound 28)
[0485]
(3S,5R)-3-Triethylsiloxy-5-(2,3-dihydroxyl)propyl-7-oxa-8-cholesten-
-6-one (High polar compound; 5.0 mg, 0.0085 mmol) obtained in the
Step 1 of Example 21 was dissolved in a mixed solvent of ethanol
(0.9 mL) and water (0.15 mL), and then sodium periodate (7.2 mg,
0.034 mmol) was added thereto at room temperature, followed by
stirring at 40.degree. C. for 1 hour. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure. The resulting residue was
dissolved in ethanol (1.0 mL), and sodium borohydride (0.6 mg,
0.0169 mmol) was added at 0.degree. C., followed by stirring for 40
minutes. A saturated aqueous ammonium chloride solution was added
to the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain the title compound
(2.4 mg, 64%).
[0486] ESI-MS: m/z 447[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.79 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.3 Hz, 3H), 0.98-2.34 (m, 28H), 2.57-2.67 (m,
1H), 3.64-3.71 (m, 2H), 3.82-3.94 (m, 1H).
EXAMPLE 23
(3S,5S)-3-Hydroxy-5-methyl-7-oxa-8-cholesten-6-one (Compound
29)
[0487] Step 1; By using
(3S,5S)-3-triethylsiloxy-7-oxa-8-cholesten-6-one (30 mg, 0.058
mmol) obtained in the Step 1 of Example 19, silver oxide (40 mg,
0.17 mmol), iodomethane (36 .mu.L, 0.58 mmol) and potassium iodide
(29 mg, 0.17 mmol),
(3S,5S)-3-triethylsiloxy-5-methyl-7-oxa-8-cholesten-6-one (8.9 mg,
29%) was obtained in the same manner as Example 17.
[0488] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.59 (q, J=8.1 Hz,
6H), 0.67 (s, 3H), 0.85 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H),
0.93 (d, J=6.3 Hz, 3H), 0.96 (t, J=8.1 Hz, 9H), 1.01-1.82 (m, 24H),
1.91-2.16 (m, 5H), 2.28-2.38 (m, 1H), 3.78-3.90 (m, 1H).
[0489] Step 2; Compound 29 (3.6 mg, 55%) was obtained by using
(3S,5S)-3-triethylsiloxy-5-methyl-7-oxa-8-cholesten-6-one (8.9 mg,
0.017 mmol) obtained in the Step 1 in the same manner as the Step 4
of Example 19.
[0490] ESI-MS: m/z 417[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 1.00-1.80 (m, 24H), 1.85-2.16 (m,
5H), 2.28-2.38 (m, 1H), 3.82-3.94 (m, 1H).
EXAMPLE 24
(3S,5S)-3-(2,3-Dihydroxyl)propoxy-7-oxa-8-cholesten-6-one (Compound
35)
[0491] Step 1; Compound 2 (230 mg, 0.571 mmol) obtained in Example
2 was dissolved in dichloromethane (6.0 mL), and
O-allyl-2,2,2-trichloroacetimidate (130 .mu.L, 0.857 mmol) and
boron trifluoride-ethyl ether complex (36.1 .mu.L, 0.286 mmol) were
added thereto at 0.degree. C., followed by stirring for 3 hours.
Subsequently, O-allyl-2,2,2-trichloroacetimidate (43.3 .mu.L, 0.286
mmol) and boron trifluoride-ethyl ether complex (18.1 .mu.L, 0.143
mmol) were further added thereto, followed by stirring for 4 hours.
A saturated aqueous sodium bicarbonate solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was washed with saturated brine, dried over
anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain
(3S,5S)-3-allyloxy-7-oxa-8-cholesten-6-one (145 mg, 57%).
[0492] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H),
0.97 (s, 3H), 1.01-1.60 (m, 15H), 1.66-1.80 (m, 2H), 1.90-2.47 (m,
8H), 3.23-3.35 (m, 1H), 3.96-4.17 (m, 2H), 5.14-5.32 (m, 2H),
5.84-6.00 (m, 1H).
[0493] Step 2; (3S,5S)-3-Allyloxy-7-oxa-8-cholesten-6-one (71 mg,
0.16 mmol) obtained in the Step 1 was dissolved in a mixed solvent
of tert-butyl alcohol (3.0 mL), THF (0.3 mL) and water (0.3 mL),
and 4-methylmorpholin-N-oxide (9.4 mg, 0.081 mmol) and 2.5% of an
osmium tetroxide/tert-butyl alcohol solution (100 .mu.L, 0.0081
mmol) were added thereto at room temperature, followed by stirring
for 90 minutes. A saturated aqueous sodium thiosulfate solution was
added to the reaction mixture, followed by extraction with ethyl
acetate twice. The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 35 (16 mg,
21%).
[0494] ESI-MS: m/z 477[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.3 Hz, 3H), 0.97 (s, 3H), 1.01-1.79 (m, 17H),
1.91-2.47 (m, 8H), 2.55-2.75 (br s, 1H), 3.23-3.35 (m, 1H),
3.49-3.75 (m, 4H), 3.81-3.90 (m, 1H).
EXAMPLE 25
(3S,5S)-3-(2-Hydroxyl)ethoxy-7-oxa-8-cholesten-6-one (Compound
31)
[0495] Step 1; Compound 35 (60.0 mg, 0.126 mmol) obtained in the
Step 2 of Example 24 was dissolved in a mixed solvent of ethanol
(3.0 mL) and water (1.0 mL), and sodium periodate (80.8 mg, 0.378
mmol) was added thereto at room temperature, followed by stirring
for 30 minutes. A saturated aqueous sodium thiosulfate solution was
added to the reaction mixture, followed by extraction with ethyl
acetate twice. The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
silica gel column chromatography to obtain
(3S,5S)-3-(formyl)methoxy-7-oxa-8-cholesten-6-one (46.6 mg,
83%).
[0496] Step 2; (3S,5S)-3-(Formyl)methoxy-7-oxa-8-cholesten-6-one
(11.0 mg, 0.0247 mmol) obtained in the Step 1 was dissolved in
ethanol (1.0 mL), and sodium borohydride (0.9 mg, 0.239 mmol) was
added thereto at 0.degree. C., followed by stirring for 15 minutes.
A saturated aqueous ammonium chloride solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was washed with saturated brine, dried over
anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 31 (7.2
mg, 67%).
[0497] ESI-MS: m/z 447[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.9 Hz, 3H), 0.97 (s, 3H), 1.00-2.47 (m, 25H),
3.24-3.36 (m, 1H), 3.48-3.77 (m, 4H).
EXAMPLE 26
(1R,2S,3S,5R)-1,2,3-Trihydroxy-7-oxa-8-cholesten-6-one (Compound
33) and (1R,2S,3R,5S)-1,2,3-trihydroxy-7-oxa-8-cholesten-6-one
(Compound 32)
[0498] Compound 18 (510 mg, 1.18 mmol) obtained in Example 15 was
dissolved in THF (10 mL), and sodium triacetoxyborohydride (750 mg,
3.54 mmol) was added thereto at room temperature, followed by
stirring for 4 hours. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative high-speed liquid chromatography to
obtain compound 33 (154 mg, 30%) and compound 32 (180 mg, 35%).
[0499] Compound 33; ESI-MS: m/z 435[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=7.0 Hz, 3H),
0.88 (d, J=7.0 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.99 (s, 3H),
1.00-1.60 (m, 12H), 1.68-2.24 (m, 5H), 2.36-2.57 (m, 3H), 2.82-2.95
(br s, 2H), 3.19 (dd, J=3.6, 13.2 Hz, 1H), 3.35-3.42 (m, 1H),
3.64-3.72 (m, 1H), 3.82-3.90 (m, 1H), 4.15-4.22 (m, 1H).
[0500] Compound 32; ESI-MS: m/z 435[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=7.0 Hz, 3H),
0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.97-1.76 (m, 18H),
1.90-2.40 (m, 5H), 2.50-2.72 (m, 2H), 3.09 (dd, J=3.3, 12.8 Hz,
1H), 3.62-3.69 (m, 1H), 3.81-3.95 (m, 2H).
EXAMPLE 27
(1R,3aR,5aS,10aS,12aR)-10a,12a-Dimethyl-1-[(R)-6-methylheptan-2-yl]-1,2,3,-
3a,5a,6,9,10,10a,11,12,12a-dodecahydro-8-azacyclohepta[c]cyclopenta[h]chro-
men-5,7-dione (Compound 34)
[0501] Compound 3 (41.5 mg, 0.0999 mmol) obtained in Example 3 was
dissolved in 1,4-dioxane (3.0 mL), and thionyl chloride (21.9
.mu.L, 0.300 mmol) was added thereto at room temperature, followed
by stirring for 1 hour. A saturated aqueous sodium bicarbonate
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain the
title compound (11.6 mg, 28%).
[0502] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H),
0.98-1.83 (m, 18H), 1.91-2.40 (m, 5H), 2.59-2.79 (m, 2H), 3.05-3.22
(m, 2H), 3.33-3.46 (m, 1H), 6.48-6.55 (br s, 1H).
EXAMPLE 28
(3S,5S)-3-(2-N-Methylamino)ethoxy-7-oxa-8-cholesten-6-one (Compound
36)
[0503] (3S,5S)-3-(Formyl)methoxy-7-oxa-8-cholesten-6-one (3.9 mg,
0.0088 mmol) obtained in the Step 1 of Example 25 was dissolved in
a mixed solvent of THF (0.5 mL) and acetic acid (0.025 mL), and 2.0
mol/L of a methylamine/THF solution (13 .mu.L, 0.026 mmol) and
sodium triacetoxyborohydride (3.7 mg, 0.018 mmol) were added
thereto at 0.degree. C., followed by stirring at room temperature
for 30 minutes. A saturated aqueous sodium bicarbonate solution was
added to the reaction mixture, followed by extraction with
chloroform four times. The organic layer was dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (2.0 mg, 50%).
[0504] ESI-MS: m/z 460[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.96 (s, 3H), 1.01-1.60 (m, 16H),
1.65-1.79 (m, 2H), 1.91-2.42 (m, 7H), 2.45 (s, 3H), 2.71-2.76 (m,
2H), 3.19-3.30 (m, 1H), 3.52-3.59 (m, 1H), 3.63-3.71 (m, 1H).
EXAMPLE 29
(3S,5S)-3-[2-(4-Morphoryl)]ethoxy-7-oxa-8-cholesten-6-one (Compound
37)
[0505] By using (3S,5S)-3-(formyl)methoxy-7-oxa-8-cholesten-6-one
(12.6 mg, 0.0283 mmol) obtained in the Step 1 of Example 25 and
morpholine (12.4 .mu.L, 0.142 mmol), the title compound (10.0 mg,
69%) was obtained in the same manner as Example 28.
[0506] ESI-MS: m/z 516[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.96 (s, 3H), 1.00-1.57 (m, 16H),
1.63-1.80 (m, 2H), 1.86-2.62 (m, 13H), 3.18-3.30 (m, 1H), 3.54-3.75
(m, 6H).
EXAMPLE 30
(3S,5S)-3-(2-Amino)ethoxy-7-oxa-8-cholesten-6-one (Compound 38)
[0507] Step 1; By using compound 31 (31.5 mg, 0.705 mmol) obtained
in the Step 2 of Example 25,
(3S,5S)-3-(2-azido)ethoxy-7-oxa-8-cholesten-6-one (25.5 mg, 77%)
was obtained in the same manner as the Step 2 of Example 10.
[0508] Step 2; Compound 38 (18.3 mg, 76%) was obtained by using
(3S,5S)-3-(2-azido)ethoxy-7-oxa-8-cholesten-6-one (25.5 mg, 0.0541
mmol) obtained in the Step 1 in the same manner as the Step 3 of
Example 10.
[0509] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H),
0.97 (s, 3H), 1.01-1.57 (m, 16H), 1.64-1.80 (m, 2H), 1.92-2.48 (m,
7H), 2.82-2.88 (m, 2H), 3.20-3.32 (m, 1H), 3.44-3.52 (m, 1H),
3.54-3.65 (m, 1H).
EXAMPLE 31
(3S,5S)-3-(2-N-Acetylamino)ethoxy-7-oxa-8-cholesten-6-one (Compound
39)
[0510] Compound 38 (4.6 mg, 0.010 mmol) obtained in the Step 2 of
Example 30 was dissolved in dichloromethane (0.5 mL), and acetic
anhydride (2.9 .mu.L, 0.031 mmol) and triethylamine (4.3 .mu.L,
0.031 mmol) were added thereto at room temperature, followed by
stirring for 3 hours. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with chloroform twice. The organic layer was dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (3.7 mg, 74%).
[0511] ESI-MS: m/z 488[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.0
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.97 (s, 3H), 1.01-1.60 (m, 16H),
1.65-1.78 (m, 2H), 1.90-2.41 (m, 10H), 3.20-3.31 (m, 1H), 3.39-3.49
(m, 2H), 3.52-3.62 (m, 2H), 5.79-5.84 (br s, 1H).
EXAMPLE 32
(3S,5S)-3-(2-N-Methanesulfonylamino)ethoxy-7-oxa-8-cholesten-6-one
(Compound 40)
[0512] By using compound 38 (4.1 mg, 0.010 mmol) obtained in the
Step 2 of Example 30 and methanesulfonyl chloride (1.1 .mu.L, 0.014
mmol), the title compound (3.8 mg, 79%) was obtained in the same
manner as Example 31.
[0513] ESI-MS: m/z 524[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.96 (s, 3H), 1.01-1.60 (m, 16H),
1.66-1.78 (m, 2H), 1.90-2.44 (m, 7H), 2.99 (s, 3H), 3.25-3.37 (m,
3H), 3.54-3.62 (m, 1H), 3.65-3.73 (m, 1H), 4.60-4.70 (br s,
1H).
EXAMPLE 33
(3S,5S)-3-(4-Pyridyl)oxy-7-oxa-8-cholesten-6-one (Compound 41)
[0514] Compound 105 (29 mg, 0.072 mmol) obtained in the Step 1 of
Example 10, 4-hydroxypyridine (21 mg, 0.22 mmol) and
triphenylphosphine (76 mg, 0.29 mmol) were dissolved in toluene
(1.0 mL), and 2.2 mol/L of an diethyl azodicarboxylate/toluene
solution (66 .mu.L, 0.14 mmol) was added thereto at room
temperature, followed by stirring for 3 hours. The residue obtained
by concentration under reduced pressure was purified by silica gel
column chromatography to obtain the title compound (5.2 mg,
15%).
[0515] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.69 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.6 Hz, 3H),
0.97-1.85 (m, 21H), 1.90-2.55 (m, 7H), 4.25-4.41 (m, 1H), 6.77 (dd,
J=1.5, 4.8 Hz, 2H), 8.41 (dd, J=1.5, 4.8 Hz, 2H).
EXAMPLE 34
(3S,5S)-3-(3-Pyridyl)oxy-7-oxa-8-cholesten-6-one (Compound 42)
[0516] By using compound 105 (46.3 mg, 0.115 mmol) obtained in the
Step 1 of Example 10 and 3-hydroxypyridine (21.9 mg, 0.230 mmol),
the title compound (12.4 mg, 22%) was obtained in the same manner
as Example 33. ESI-MS: m/z 480[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.69 (s, 3H), 0.87 (d, J=6.6 Hz, 3H), 0.87 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.97-1.83 (m, 20H), 1.91-2.39 (m,
6H), 2.43-2.56 (m, 2H), 4.23 (m, 1H), 7.17-7.21 (m, 2H), 8.22 (m,
1H), 8.30 (m, 1H).
EXAMPLE 35
(3S,5S)-3-(2-N-Phenylcarbamoylamino)ethoxy-7-oxa-8-cholesten-6-one
(Compound 43)
[0517] By using compound 38 (5.5 mg, 0.012 mmol) obtained in the
Step 2 of Example 30 and phenyl isocyanate (1.6 .mu.L, 0.015 mmol),
the title compound (4.1 mg, 59%) was obtained in the same manner as
Example 31.
[0518] ESI-MS: m/z 565[M+1-1].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.68 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.96 (s, 3H), 1.01-1.60 (m, 17H),
1.66-1.78 (m, 2H), 1.90-2.44 (m, 7H), 2.99 (s, 3H), 3.25-3.37 (m,
3H), 3.57 (m, 1H), 3.68 (m, 1H), 4.65 (br s, 1H).
EXAMPLE 36
(3S,5S)-3-(2-Hydroxyl)propoxy-7-oxa-8-cholesten-6-one (1:1
diastereomer mixture, Compound 44) and
(3S,5S)-3-(3-hydroxyl)propoxy-7-oxa-8-cholesten-6-one (Compound
45)
[0519] Step 1; (3S,5S)-3-Allyloxy-7-oxa-8-cholesten-6-one (50.8 mg,
0.115 mmol) obtained in the Step 1 of Example 24 was dissolved in a
mixed solvent of DMF (2.1 mL) and water (0.3 mL), and palladium
chloride (10.2 mg, 0.0574 mmol) and cupric acetate monohydrate
(22.9 mg, 0.115 mmol) were added thereto at room temperature,
followed by stirring overnight. Subsequently, palladium chloride
(10.2 mg, 0.0574 mmol) and cupric acetate monohydrate (22.9 mg,
0.115 mmol) were added, followed by stirring overnight. To the
reaction mixture, 10% aqueous ammonia was added, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain a
mixture (14.8 mg, 28%) of
(3S,5S)-3-(2-oxo)propoxy-7-oxa-8-cholesten-6-one and
(3S,5S)-3-(2-formyl)ethoxy-7-oxa-8-cholesten-6-one.
[0520] Step 2; A mixture (15 mg, 0.032 mmol) of
(3S,5S)-3-(2-oxo)propoxy-7-oxa-8-cholesten-6-one and
(3S,5S)-3-(2-formyl)ethoxy-7-oxa-8-cholesten-6-one obtained in the
Step 1 was dissolved in methanol (1.0 mL), and sodium borohydride
(1.2 mg, 0.032 mmol) was added thereto at 0.degree. C., followed by
stirring for 10 minutes. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain
compound 44 (13 mg, 84%) and compound 45 (1.8 mg, 12%).
[0521] Compound 44; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s,
3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3
Hz, 3H), 0.96 (s, 3H), 1.01-1.78 (m, 21H), 1.90-2.46 (m, 7H),
3.16-3.37 (m, 2H), 3.48 (dd, J=3.3, 9.2 Hz, 0.5H), 3.57 (dd, J=3.3,
9.2 Hz, 0.5H), 3.89 (m, 1H).
[0522] Compound 45; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s,
3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3
Hz, 3H), 0.97 (s, 3H), 1.01-1.57 (m, 18H), 1.65-1.87 (m, 4H),
1.91-2.50 (m, 7H), 3.20-3.31 (m, 1H), 3.59-3.67 (m, 1H), 3.73-3.81
(m, 1H).
EXAMPLE 37
(3S,5S)-3-Glucopyranosiloxy-7-oxa-8-cholesten-6-one (1:1
diastereomer mixture, Compound 46)
[0523] Step 1; By using compound 2 (15.0 mg, 0.0373 mmol) obtained
in Example 2 and 2,3,4,6-tetra-O-benzyl-.alpha.-D-glucopyranosyl
trichloroacetimidate (38.3 mg, 0.0559 mmol),
(3S,5S)-3-(2,3,4,6-tetra-O-benzyloxyglucopyranosyl)oxy-7-oxa-8-cholesten--
6-one (1:1 diastereomer mixture) (15.4 mg, 45%) was obtained in the
same manner as the Step 1 of Example 24.
[0524] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H),
0.97 (s, 3H), 1.01-1.79 (m, 18H), 1.84-2.50 (m, 7H), 3.44-4.00 (m,
7H), 4.40-5.01 (m, 9H), 7.11-7.40 (m, 20H).
[0525] Step 2;
(3S,5S)-3-(2,3,4,6-Tetra-O-benzyloxyglucopyranosyl)oxy-7-oxa-8-cholesten--
6-one (15.4 mg, 0.0167 mmol) obtained in the Step 1 was dissolved
in ethanol (1.0 mL), and 10% palladium-carbon (10.0 mg) was added
thereto at room temperature, followed by stirring for 6 hours under
a hydrogen atmosphere. The reaction mixture was filtered through a
celite, and the filtrate was concentrated under reduced pressure to
yield a residue. The residue was purified by preparative thin layer
chromatography to obtain compound 46 (4.4 mg, 47%).
[0526] ESI-MS: m/z 565[M+H].sup.+, .sup.1H NMR (DMSO-d.sub.6)
.delta.(ppm): 0.64 (s, 1.5H), 0.73 (s, 1.5H), 0.83-0.93 (m, 12H),
1.00-2.60 (m, 25H), 2.85-3.69 (m, 7H), 4.22-4.95 (m, 5H).
EXAMPLE 38
(3S,5S)-3-Benzyloxy-7-oxa-8-cholesten-6-one (Compound 47)
[0527] By using compound 2 (48.0 mg, 0.119 mmol) obtained in
Example 2 and O-benzyl 2,2,2-trichloroacetamidate (33.2 .mu.L,
0.179 mmol), the title compound (20.0 mg, 34%) was obtained in the
same manner as the Step 1 of Example 24.
[0528] ESI-MS: m/z 493[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.3 Hz, 3H), 0.97 (s, 3H), 1.01-1.80 (m, 18H),
1.84-2.37 (m, 6H), 2.45-2.55 (m, 1H), 3.30-3.41 (m, 1H), 4.53 (d,
J=11.7 Hz, 1H), 4.65 (d, J=11.7 Hz, 1H), 7.12-7.38 (m, 5H).
EXAMPLE 39
(2R,5S)-2-Hydroxy-7-oxa-8-cholesten-3,6-dione (Compound 48) and
(4S,5R)-4-hydroxy-7-oxa-8-cholesten-3,6-dione (Compound 51)
[0529] Compound 1 (200 mg, 0.500 mmol) obtained in Example 1 was
dissolved in a mixed solvent of acetonitrile (3.0 mL) and
dichloromethane (1.5 mL), and hexamethyldisilazane (315 .mu.L, 1.50
mmol), sodium iodide (150 mg, 0.999 mmol) and chlorotrimethylsilane
(95.0 .mu.L, 0.749 mmol) were added thereto at 0.degree. C.,
followed by stirring at room temperature for 1 hour. Subsequently,
chlorotrimethylsilane (31.7 .mu.L, 0.250 mmol) was further added
thereto, followed by stirring for 30 minutes. A saturated aqueous
sodium bicarbonate solution was added to the reaction mixture,
followed by extraction with ethyl acetate twice. The organic layer
was washed with saturated brine, dried over anhydrous magnesium
sulfate, and concentrated under reduced pressure. The resulting
residue was dissolved in diethyl ether (4.0 mL), and
4-methylmorpholin-N-oxide (87.7 mg, 0.749 mmol), 4% of an aqueous
solution of osmium tetroxide (153 .mu.L, 0.0250 mmol) and water
(2.0 mL) were added thereto at room temperature, followed by
stirring for 4 hours. A saturated aqueous sodium thiosulfate
solution was added to the reaction mixture, followed by extraction
with ethyl acetate twice. The organic layer was washed with
saturated brine, dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain compound
48 (121 mg, 58%) and compound 51 (3.3 mg, 2%).
[0530] Compound 48; ESI-MS: m/z 417[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.70 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98-1.81 (m, 17H),
1.92-2.47 (m, 6H), 2.66-2.85 (m, 2H), 2.95 (dd, J=3.3, 13.6 Hz,
1H), 3.50-3.53 (m, 1H), 4.24-4.34 (m, 1H).
[0531] Compound 51; ESI-MS: m/z 417[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.70 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98-1.87 (m, 17H),
1.92-2.40 (m, 6H), 2.57-2.70 (m, 3H), 3.74-3.82 (br s, 1H), 4.54
(d, J=10.6 Hz, 1H).
EXAMPLE 40
(2R,3S,5S)-2,3-Dihydroxy-7-oxa-8-cholesten-6-one (Compound 50) and
(2R,3R,5S)-2,3-hydroxy-7-oxa-8-cholesten-6-one (Compound 49)
[0532] Compound 48 (3.1 mg, 0.0074 mmol) obtained in Example 39 was
dissolved in a mixed solvent of methanol (0.5 mL) and
dichloromethane (0.25 mL), and sodium borohydride (0.6 mg, 0.015
mmol) was added thereto at 0.degree. C., followed by stirring for
10 minutes. A saturated aqueous ammonium chloride solution was
added to the reaction mixture, followed by extraction with ethyl
acetate twice. The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 50 (0.5
mg, 16%) and compound 49 (2.1 mg, 67%).
[0533] Compound 50; ESI-MS: m/z 419[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98 (s, 3H),
1.01-2.39 (m, 22H), 2.88 (dd, J=3.3, 12, 8 Hz, 1H), 3.80 (m, 1H),
4.09 (m, 1H).
[0534] Compound 49; ESI-MS: m/z 419[M+H].sup.+, .sup.1H NMR
(CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H),
0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.98-1.78 (m, 17H),
1.91-2.41 (m, 8H), 2.51 (dd, J=3.3, 12, 8 Hz, 1H), 3.44 (m, 1H),
3.65 (m, 1H).
EXAMPLE 41
(2R,3R,5S)-2-Hydroxy-3-methoxymethoxy-7-oxa-8-cholesten-6-one
(Compound 52)
[0535] Step 1; Compound 48 (570 mg, 1.37 mmol) obtained in Example
39 was dissolved in dichloromethane (10 mL), and imidazole (186 mg,
2.74 mmol) and chiorotriethylsilane (344 .mu.L, 2.05 mmol) were
added thereto at room temperature, followed by stirring for 5
hours. A saturated aqueous ammonium chloride solution was added to
the reaction mixture, followed by extraction with chloroform twice.
The organic layer was dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain
(2R,5S)-2-triethylsiloxy-7-oxa-8-cholesten-3,6-dione (676 mg,
93%).
[0536] ESI-MS: m/z 531[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.63 (q, J=8.1 Hz, 6H), 0.70 (s, 3H), 0.86 (d, J=6.6
Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.3 Hz, 3H), 0.96 (t,
J=8.1 Hz, 9H), 1.00-1.57 (m, 16H), 1.69-1.80 (m, 2H), 1.93-2.41 (m,
6H), 2.55-2.66 (m, 1H), 2.80 (m, 1H), 4.31 (m, 1H).
[0537] Step 2; (2R,5S)-2-Triethylsiloxy-7-oxa-8-cholesten-3,6-dione
(37.2 mg, 0.0701 mmol) obtained in the Step 1 was dissolved in THF
(1.0 mL), and sodium triacetoxyborohydride (29.7 mg, 0.140 mmol)
was added thereof at room temperature, followed by stirring
overnight. A saturated aqueous sodium bicarbonate solution was
added to the reaction mixture, followed by extraction with ethyl
acetate twice. The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
silica gel column chromatography to obtain
(2R,3R,5S)-2-triethylsiloxy-3-hydroxy-7-oxa-8-cholesten-6-one (30.9
mg, 83%).
[0538] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.63 (q, J=8.1 Hz,
6H), 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H),
0.93 (d, J=6.6 Hz, 3H), 0.98 (t, J=8.1 Hz, 9H), 1.00-1.85 (m, 18H),
1.92-2.23 (m, 4H), 2.30-2.53 (m, 4H), 3.40 (m, 1H), 3.62 (m,
1H).
[0539] Step 3;
(2R,3R,5S)-2-Triethylsiloxy-3-hydroxy-7-oxa-8-cholesten-6-one (5.9
mg, 0.011 mmol) obtained in the Step 2 was dissolved in
dichloromethane (0.5 mL), and N,N-diisopropylethylamine (77.1
.mu.L, 0.442 mmol) and chloromethylmethyl ether (13.5 .mu.L, 0.177
mmol) were added thereto at room temperature, followed by stirring
overnight. A saturated aqueous ammonium chloride solution was added
to the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain
(2R,3R,5S)-2-triethylsiloxy-3-methoxymethoxy-7-oxa-8-cholesten-6-one
(4.6 mg, 72%).
[0540] .sup.1H NMR(CDCl.sub.3) .delta.(ppm): 0.61 (q, J=8.1 Hz,
6H), 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H),
0.93 (d, J=6.6 Hz, 3H), 0.96 (t, J=8.1 Hz, 9H), 0.99 (s, 3H),
1.02-1.84 (m, 15H), 1.92-2.48 (m, 8H), 3.37 (m, 1H), 3.40 (s, 3H),
3.72 (m, 1H), 4.68 (d, J=6.6 Hz, 1H), 4.80 (d, J=6.6 Hz, 1H).
[0541] Step 4; By using
(2R,3R,5S)-2-triethylsiloxy-3-methoxymethoxy-7-oxa-8-cholesten-6-one
(4.6 mg, 0.00797 mmol) obtained in the Step 3, compound 52 (3.3 mg,
89%) was obtained in the same manner as the Step 4 of Example
19.
[0542] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.6 Hz, 3H),
0.99-1.78 (m, 18H), 1.92-2.48 (m, 8H), 3.29 (m, 1H), 3.44 (s, 3H),
3.58 (br s, 1H), 3.68 (m, 1H), 4.74 (d, J=7.0 Hz, 1H), 4.78 (d,
J=7.0 Hz, 1H).
EXAMPLE 42
(2R,3R,5S)-2,3-Dihydroxy-7-oxa-8-cholesten-6-one (Compound 53)
[0543] Step 1;
(2R,3R,5S)-2-Triethylsiloxy-3-hydroxy-7-oxa-8-cholesten-6-one (30.9
mg, 0.0580 mmol) obtained in the Step 2 of Example 41 was dissolved
in dichloromethane (1.0 mL), and 2,6-lutidine (20.2 .mu.L, 0.174
mmol) and tert-butyldimethyl silyl trifluoromethanesulfonate (24.0
.mu.L, 0.104 mmol) were added at 0.degree. C., followed by stirring
at room temperature for 15 minutes. A saturated aqueous sodium
bicarbonate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain
(2R,3R,5S)-2-triethylsiloxy-3-(tert-butyldimethylsiloxy)-7-oxa-8-choleste-
n-6-one (20.5 mg, 55%).
[0544] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.07 (s, 6H), 0.62
(q, J=8.1 Hz, 6H), 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d,
J=6.6 Hz, 3H), 0.90 (s, 9H), 0.93 (d, J=6.3 Hz, 3H), 0.97 (t, J=8.1
Hz, 9H), 1.00 (s, 3H), 1.02-1.85 (m, 16H), 1.91-2.47 (m, 7H), 3.41
(m, 1H), 3.63 (m, 1H).
[0545] Step 2; By using
(2R,3R,5S)-2-triethylsiloxy-3-(tert-butyldimethylsiloxy)-7-oxa-8-choleste-
n-6-one (7.5 mg, 0.012 mmol) obtained in the Step 1,
(2R,3R,5S)-2-hydroxy-3-(tert-butyldimethylsiloxy)-7-oxa-8-cholesten-6-one
(5.3 mg, 86%) was obtained in the same manner as the Step 3 of
Example 19.
[0546] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.11 (s, 3H), 0.12
(s, 3H), 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz,
3H), 0.91 (s, 9H), 0.93 (d, J=6.3 Hz, 3H), 0.98-1.78 (m, 18H),
1.92-2.10 (m, 3H), 2.14-2.40 (m, 4H), 2.49 (dd, J=3.3, 12.8 Hz,
1H), 3.39 (m, 1H), 3.61 (m, 1H).
[0547] Step 3;
(2R,3R,5S)-2-Hydroxy-3-(tert-butyldimethylsiloxy)-7-oxa-8-cholesten-6-one
(12.0 mg, 0.0225 mmol) obtained in the Step 2 was dissolved in
dichloromethane (1.0 mL), and 4-methylmorpholine N-oxide (5.3 mg,
0.045 mmol) and tetrapropylammonium perruthenate (1.6 mg, 0.045
mmol) were added thereto at room temperature, followed by stirring
for 2 hours at room temperature. A saturated aqueous sodium
thiosulfate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was washed
with saturated brine, dried over anhydrous magnesium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain
(3R,5S)-3-(tert-butyldimethylsiloxy)-7-oxa-8-cholesten-2,6-dione
(11.0 mg, 92%).
[0548] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.03 (s, 3H), 0.14
(s, 3H), 0.66 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz,
3H), 0.90 (s, 9H), 0.93 (d, J=6.3 Hz, 3H), 0.94 (s, 3H), 1.01-1.59
(m, 14H), 1.76 (m, 1H), 1.93-2.18 (m, 5H), 2.38 (m, 1H), 2.67 (m,
1H), 2.98 (dd, J=3.6, 12.9 Hz, 1H), 4.22 (m, 1H).
[0549] Step 4;
(3R,5S)-3-(Tert-butyldimethylsiloxy)-7-oxa-8-cholesten-2,6-dione
(7.0 mg, 0.013 mmol) obtained in the Step 3 was dissolved in a
mixed solvent of methanol (0.5 mL) and dichloromethane (0.25 mL),
and sodium borohydride (1.5 mg, 0.040 mmol) was added thereto at
0.degree. C., followed by stirring for 40 minutes at room
temperature. A saturated aqueous ammonium chloride solution was
added to the reaction mixture, followed by extraction with ethyl
acetate twice. The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated under
reduced pressure. Compound 53 (2.5 mg, 45%) was obtained by using
the resulting residue in the same manner as the Step 4 of Example
39.
[0550] ESI-MS: m/z 419[M+H].sup.+, 1H NMR (CDCl.sub.3)
.delta.(ppm): 0.67 (s, 3H), 0.86 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (d, J=6.6 Hz, 3H), 0.98-1.78 (m, 19H), 1.85-2.43 (m,
9H), 3.72 (m, 1H), 4.07 (m, 1H).
EXAMPLE 43
(3R,5S)-3-Hydroxy-7-oxa-8-cholesten-2,6-dione (Compound 54)
[0551] The title compound (2.8 mg, 70%) was obtained by using
compound 52 (4.0 mg, 0.0087 mmol) obtained in the Step 4 of Example
41 in the same manner as the Step 3 of Example 42.
[0552] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.67 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H),
0.94 (s, 3H), 0.98-1.81 (m, 15H), 1.93-2.18 (m, 5H), 2.35 (m, 1H),
2.77 (m, 1H), 2.98 (dd, J=4.0, 12.9 Hz, 1H), 3.41 (s, 3H), 4.27 (m,
1H), 4.74 (s, 2H).
EXAMPLE 44
(2R,5S)-2-Hydroxy-3-hydroxyimino-7-oxa-8-cholesten-6-one (Compound
55)
[0553] The title compound (10.2 mg, 68%) was obtained as a single
geometric isomer by using compound 48 (14.6 mg, 0.0350 mmol)
obtained in Example 39 in the same manner as Example 3.
[0554] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.69 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.93 (d, J=6.3 Hz, 3H),
0.94 (s, 3H), 0.98-1.82 (m, 16H), 1.91-2.40 (m, 6H), 2.57 (dd,
J=4.3, 13.5 Hz, 1H), 2.98 (dd, J=4.0, 12.9 Hz, 1H), 3.78 (dd,
J=4.6, 15.8 Hz, 1H), 4.39 (m, 1H).
EXAMPLE 45
7-Oxa-1,4,8-cholestatrien-3,6-dione (Compound 57)
[0555] Compound 4 (5.8 mg, 0.015 mmol) obtained in Example 4 and
N-fluorobenzenesulfonimide (6.0 mg, 0.019 mmol) were dissolved in
THF (0.5 mL), and 1.0 mol/L of a lithium
bis(trimethylsilyl)amide/THF solution (18.9 .mu.L, 0.0189 mmol) was
added thereto at 0.degree. C., followed by stirring for 40 minutes.
A saturated aqueous ammonium chloride solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was washed with saturated brine, dried over
anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain the title compound
(1.9 mg, 33%).
[0556] ESI-MS: m/z 397[M+H].sup.+, .sup.1H NMR (CDCl.sub.3)
.delta.(ppm): 0.74 (s, 3H), 0.85 (d, J=6.6 Hz, 3H), 0.86 (d, J=6.6
Hz, 3H), 0.92 (d, J=6.3 Hz, 3H), 0.96-2.53 (m, 21H), 6.35 (m, 1H),
6.90 (m, 1H), 7.04 (d, J=10.2 Hz, 1H).
EXAMPLE 46
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6-one (Compound 56)
[0557] Compound P9 (48.5 mg, 0.105 mmol) obtained in Reference
Example 9 was dissolved in acetic anhydride (3 mL), and sodium
acetate (200 mg, 2.44 mmol) was added thereto, followed by stirring
at 105.degree. C. for 2 hours. After being left to cool, water was
added to the reaction mixture, followed by extraction with ethyl
acetate (20 mL.times.3). The organic layer was washed with water
and saturated brine, and dried over anhydrous sodium sulfate. The
residue obtained by concentration was purified by silica gel column
chromatography (5 to 20% ethyl acetate/n-hexane) to obtain the
title compound (16.2 mg, 35%).
[0558] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66 (2H, m),
3.53 (1H, m), 3.38 (3H, s), 2.37-1.01 (22H, m), 1.25 (3H, s), 0.97
(3H, s), 0.93 (3H, d, J=6.4 Hz), 0.87 (3H, d, J=6.6 Hz), 0.87 (3H,
d, J=6.6 Hz), 0.68 (3H, s).
EXAMPLE 47
20-(Acetyloxymethyl)-3-(methoxymethyloxy)-7-oxa-pregn-8(9)-en-6-one
(Compound 58)
[0559] Compound P17 (45.3 mg, 0.100 mmol) obtained in Reference
Example 17 was treated with sodium acetate (250 mg, 3.05 mmol) and
acetic anhydride (1 mL) in the same manner as Example 46 to obtain
the title compound (8.3 mg, 19%).
[0560] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.71 (2H, m), 4.11
(1H, m), 3.72 (1H, m), 3.50 (1H, m), 3.38 (3H, s), 2.05 (3H, s),
2.50-1.10 (18H, m), 1.00 (3H, d, J=6.6 Hz), 0.95 (3H, s), 0.65 (3H,
s).
EXAMPLE 48
20-(Acetyloxymethyl)-7-oxa-pregn-8(9)-en-6-on-3-ol (Compound
77)
[0561] Compound P58 (8.3 mg, 0.019 mmol) obtained in Example 47 was
treated with pyrimidium p-toluenesulfonate (14.4 mg, 0.0573 mmol)
and tert-butanol (1 mL) in the same manner as Example 63 to obtain
the title compound (1.2 mg, 16%).
[0562] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.08 (1H, dd, J=10.8,
3.5 Hz), 3.79 (1H, dd, J=10.8, 7.5 Hz), 3.65 (1H, m), 2.39 (1H, dd,
J=12.7, 3.3 Hz), 2.06 (3H, s), 2.33-1.25 (18H, m), 1.03 (3H, d,
J=6.6 Hz), 0.97 (3H, s), 0.70 (3H, s).
EXAMPLE 49
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6-on-22-ol (Compound
60)
[0563] Compound P24 (3.7 mg, 0.0064 mmol) obtained in Reference
Example 24 was dissolved in THF (1 mL), and a hydrogen
fluoride-pyridine complex (0.5 mL) was added thereto, follwed by
stirring at room temperature for 20 minutes. A saturated aqueous
sodium bicarbonate solution was added to the reaction mixture,
followed by extraction with chloroform. The organic layer was
washed with saturated brine, dried over anhydrous sodium sulfate,
followed by purification by silica gel preparative thin layer
chromatography (2:1 n-hexane-ethyl acetate) to obtain the title
compound (2.3 mg, 78%).
[0564] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.70 (2H, m), 3.62
(1H, m), 3.53 (1H, m), 3.38 (3H, s), 2.39-1.15 (24H, m), 0.97 (3H,
s), 0.92 (3H, d, J=7.5 Hz), 0.90 (6H, d, J=6.6 Hz), 0.69 (3H,
s).
EXAMPLE 50
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6,24-dione (Compound 61)
and 3-(methoxymethyloxy)-7-oxa-cholest-8(14)-en-6,24-dione
(Compound 92)
[0565] Compound P31 (17.7 mg, 0.0370 mmol) obtained in Reference
Example 30 was treated with sodium acetate (100 mg, 0.209 mmol) and
acetic acid (1 mL) in the same manner as Example 46 to obtain
compound 61 (4.6 mg, 27%) and compound 92 (1.8 mg, 11%).
[0566] Compound 61; .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66
(2H, m), 3.52 (1H, m), 3.38 (3H, s), 2.61 (1H, m), 2.52-1.19 (22H,
m), 1.10 (3H, d, J=7.0 Hz), 1.09 (3H, d, J=6.8 Hz), 0.97 (3H, s),
0.93 (3H, d, J=6.2 Hz), 0.67 (3H, s).
[0567] Compound 92; .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66
(2H, m), 3.53 (1H, m), 3.38 (3H, s), 2.61 (1H, m), 2.52-1.20 (22H,
m), 1.09 (6H, d, J=6.8 Hz), 0.94 (3H, s), 0.93 (3H, d, J=6.6 Hz),
0.84 (3H, s).
EXAMPLE 51
7-Oxa-cholest-8(9)-en-6,24-dion-3-ol (Compound 78)
[0568] Compound 61 (4.6 mg, 0.010 mmol) obtained in Example 50 was
treated with pyrimidium p-toluenesulfonate (25 mg, 0.10 mmol) and
tert-butanol (1 mL) in the same manner as Example 63 to obtain the
title compound (2.6 mg, 63%).
[0569] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.63 (1H, m), 2.61
(1H, m), 2.53-1.22 (23H, m), 1.09 (6H, d, J=7.0 Hz), 0.97 (3H, s),
0.93 (3H, d, J=6.4 Hz), 0.67 (3H, s).
EXAMPLE 52
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6-on-26-ol (Compound
62)
[0570] To an anhydrous THF solution (12.5 mL) containing compound
P41 (16.7 mg, 0.0270 mmol) obtained in Reference Example 40 was
added hydrogen fluoride-pyridine complex (1.0 mL), followed by
stirring at room temperature for 30 minutes. The reaction mixture
was poured into a saturated aqueous sodium bicarbonate solution,
followed by extraction with chloroform. The organic layer was
washed with saturated brine, dried over anhydrous sodium sulfate,
and concentrated to yield a residue. The residue was purified by
silica gel column chromatography (20 to 60% ethyl acetate/n-hexane)
to obtain the title compound (10.8 mg, 87%).
[0571] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66 (2H, m),
3.57-3.40 (3H, m), 3.38 (3H, s), 2.37-1.25 (26H, m), 0.97 (3H, s),
0.93 (3H, d, J=6.4 Hz), 0.91 (3H, d, J=6.4 Hz), 0.67 (3H, s).
EXAMPLE 53
7-Oxa-cholest-8(9)-en-6-on-3,26-diol (Compound 79) and
7-oxa-cholest-8(14)-en-6-on-3,26-diol (Compound 90)
[0572] To a tert-butanol solution (1 mL) containing compound 62
(1.5 mg, 0.0033 mmol) obtained in Example 52 was added pyrimidium
p-toluenesulfonate (17.5 mg, 0.0696 mmol), followed by stirring at
100.degree. C. for 3 hours. The reaction mixture was concentrated,
followed by purification by silica gel preparative thin layer
chromatography (chloroform-methanol (9:1)) to obtain compound 79
(0.6 mg, 49%) and compound 90 (0.5 mg, 41%).
[0573] Compound 79; .sup.1H NMR 8(ppm, CDCl.sub.3): 3.64 (1H, m),
3.53-3.40 (2H, m), 2.39 (1H, dd, J=12.7, 3.5 Hz), 2.33-1.00 (26H,
m), 0.97 (3H, s), 0.94 (3H, d, J=6.0 Hz), 0.92 (3H, d, J=6.6 Hz),
0.67 (3H, s).
[0574] Compound 90; .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.64 (1H,
m), 3.53-3.43 (2H, m), 2.43-1.00 (27H, m), 0.93 (3H, d, J=6.5 Hz),
0.92 (3H, s), 0.91 (3H, d, J=6.5 Hz), 0.84 (3H, s).
EXAMPLE 54
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6-on-26-al (Compound
63)
[0575] To a dichloromethane solution (2 mL) containing compound 62
(10.8 mg, 0.0234 mmol) obtained in Example 52, Molecular Sieves 4
.ANG. (200 mg), 4-methylmorpholin-N-oxide (10 mg, 0.085 mmol) and
tetrapropylammonium perruthenate (3.0 mg, 0.0085 mmol) were added,
followed by stirring at room temperature for 15 minutes. The
reaction mixture was filtered, and the filtrate was concentrated,
followed by purification by silica gel column chromatography (20 to
50% ethyl acetate/n-hexane) to obtain the title compound (4.5 mg,
42%).
[0576] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 9.61 (1H, d, J=2.0
Hz), 4.73-4.66 (2H, m), 3.52 (1H, m), 3.38 (3H, s), 2.37-1.23 (25H,
m), 1.09 (3H, d, J=7.0 Hz), 0.97 (3H, s), 0.93 (3H, d, J=6.4 Hz),
0.67 (3H, s).
EXAMPLE 55
3-(Methoxymethyloxy)-7-oxa-cholest-8(9)-en-6-on-26-oic acid
(Compound 64)
[0577] Compound 63 (4.5 mg, 0.0098 mmol) obtained in Example 54,
2-methyl-2-butene (50 .mu.L, 0.47 mmol) and sodium
dihydrogenphosphate (20 mg, 0.13 mmol) were dissolved in a mixed
solvent of tert-butanol (2 mL) and water (0.5 mL), and sodium
chlorite (80%; 20 mg, 0.18 mmol) was added thereto, followed by
stirring for 15 minutes. The reaction mixture was poured into a
saturated aqueous ammonium chloride solution, followed by
extraction with chloroform (10 mL.times.3). The organic layer was
dried over anhydrous sodium sulfate, and concentrated, followed by
purification by silica gel column chromatography (5 to 10%
methanol/chloroform) to obtain the title compound (2.9 mg,
62%).
[0578] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73-4.66 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 2.51-1.25 (26H, m), 1.18 (3H, d, J=7.0
Hz), 0.97 (3H, s), 0.93 (3H, d, J=6.6 Hz), 0.67 (3H, s).
EXAMPLE 56
7-Oxa-3-hydroxy-cholest-8(9)-en-6-on-26-acid (Compound 80)
[0579] To a tert-butanol solution containing compound 64 (2.9 mg,
0.0061 mmol) obtained in Example 55 was added pyrimidium
p-toluenesulfonate (5.0 mg, 0.020 mmol), followed by stirring at
110.degree. C. for 2.5 hours. The reaction mixture was
concentrated, followed by purification by silica gel preparative
thin layer chromatography (chloroform-methanol (9:1)) to obtain the
title compound (1.1 mg, 42%).
[0580] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.64 (1H, m),
2.48-1.25 (27H, m), 1.18 (3H, d, J=6.8 Hz), 0.95 (3H, d, J=7.0 Hz),
0.92 (3, s), 0.67 (3H, s).
EXAMPLE 57
tert-Butyl
3-(methoxymethyloxy)-20-carboxy-7-oxa-pregna-8(14)-en-6-on-22-a- te
(Compound 59)
[0581] Compound P45 (30.1 mg, 0.0627 mmol) obtained in Reference
Example 44 was treated with sodium acetate (205 mg, 2.50 mmol) and
acetic anhydride (1 mL) in the same manner as Example 46 to obtain
the title compound (8.8 mg, 30%).
[0582] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.69 (2H, m), 3.52
(1H, m), 3.38 (3H, s), 2.39-2.12 (5H, m), 1.99-1.86 (2H, m),
1.81-1.61 (2H, m), 1.56-1.45 (9H, m), 1.44 (9H, s), 1.16 (3H, d,
J=6.8 Hz), 0.97 (3H, s), 0.68 (3H, s).
EXAMPLE 58
tert-Butyl 20-carboxy-7-oxa-pregna-8(14)-en-6-on-22-ate (Compound
87)
[0583] Compound 59 (2.6 mg, 0.0056 mmol) obtained in Example 57 was
treated with pyrimidium p-toluenesulfonate (14.5 mg, 0.0577 mmol)
and tert-butanol (1 mL) in the same manner as Example 63 to obtain
the title compound (1.3 mg, 55%). .sup.1H NMR .delta.(ppm,
CDCl.sub.3): 3.63 (1H, m), 2.44 (1H, dd, J=12.5, 3.3 Hz), 2.34-2.22
(2H, m), 2.00-1.80 (2H, m), 1.76-1.60 (2H, m), 1.59-1.25 (12H, m),
1.44 (9H, s), 1.16 (3H, d, J=6.8 Hz), 0.97 (3H, s), 0.68 (3H,
s).
EXAMPLE 59
20-Carboxy-7-oxa-pregna-8(14)-en-6-on-22-acid (Compound 88)
[0584] Compound 59 (8.8 mg, 0.019 mmol) obtained in Example 57 was
dissolved in dichloromethane (2 mL), and a dichloromethane solution
(0.2 mL) of B-bromocatecholborane (9.0 mg, 0.045 mmol) was added
thereto, followed by stirring at room temperature for 69 hours.
Water was added to the reaction mixture, followed by extraction
with chloroform. The organic layer was washed with saturated brine,
dried over anhydrous sodium sulfate, followed by purification by
silica gel preparative thin layer chromatography
(chloroform-methanol (10:1)) to obtain the title compound (6.0 mg,
87%).
[0585] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.64 (1H, m),
2.58-2.21 (4H, m), 1.97-1.39 (16H, m), 1.26 (3H, d, J=7.0 Hz), 0.96
(3H, s), 0.85 (3H, s).
EXAMPLE 60
20-Carboxy-7-oxa-pregna-8(14)-en-6-on-22-acid tert-butyl
N-(2-aminoethyl)carbamate (Compound 89)
[0586] Compound 88 (1.3 mg, 0.0036 mmol) obtained in Example 59,
N-(tert-butoxycarbonyl)ethylene diamine (1.5 mg, 0.0094 mmol),
1-hydroxybenzotriazole (3.0 mg, 0.022 mmol) and triethylamine (1.0
.mu.L, 0.0072 mmol) were dissolved in dichloromethane (1.2 mL), and
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.0
mg, 0.010 mmol) was added thereto, followed by stirring at room
temperature for 20 hours. A saturated aqueous ammonium chloride
solution was added to the reaction mixture, followed by extraction
with chloroform. The organic layer was washed with saturated brine,
dried over anhydrous sodium sulfate, and concentrated to yield a
residue. The residue was purified by silica gel preparative thin
layer chromatography (chloroform-methanol (10:1)) to obtain the
title compound (1.5 mg, 83%).
[0587] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 6.18 (1H, br s), 4.89
(1H, br s), 3.65 (1H, m), 3.37-3.25 (4H, m), 2.52-2.38 (2H, m),
2.26-2.21 (3H, m), 2.12 (1H, m), 2.07-1.25 (13H, m), 1.44 (9H, s),
1.19 (3H, d, J=6.8 Hz), 0.93 (3H, s), 0.84 (3H, s).
EXAMPLE 61
7-Oxa-ergosta-8(9)-en-6-on-3-ol (Compound 76)
[0588] 7-Oxa-ergosta-8(9), 24(28)-dien-6-on-3-ol (4.0 mg, 0.0097
mmol) obtained in Reference Example 85 was dissolved in ethyl
acetate (2 mL), and 10% palladium-carbon (2.4 mg) was added
thereto, followed by stirring at room temperature for 1 hour under
a hydrogen atmosphere. The catalyst was separated by filtration,
and the filtrate was concentrated to obtain the title compound (3.6
mg, 90%).
[0589] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.63 (1H, m), 2.39
(1H, dd, 3=12.7, 3.3 Hz), 2.33-1.00 (24H, m), 0.97 (3H, s), 0.93
(3H, d, J=6.4 Hz), 0.85 (3H, d, J=6.6 Hz), 0.78 (3H, d, J=7.0 Hz),
0.77 (3H, d, J=7.0 Hz), 0.67 (3H, s).
EXAMPLE 62
3-(Acetyloxy)-7-oxa-ergosta-8(9)-en-6-one (Compound 91)
[0590] Compound P50 (24.0 mg, 0.0504 mmol) obtained in Reference
Example 49 was dissolved in acetic anhydride (1 mL), and sodium
acetate (150 mg, 1.83 mmol) was added thereto, followed by stirring
at 90.degree. C. to 95.degree. C. for 1 hour. The reaction mixture
was cooled to room temperature, followed by adding ethyl acetate (5
mL) then filtration. Water was added to the filtrate, followed by
extraction with ethyl acetate (20 mL.times.2). The organic layer
was washed with a saturated aqueous sodium bicarbonate solution and
saturated brine, and dried over anhydrous magnesium sulfate,
followed by purification by silica gel column chromatography (5 to
15% ethyl acetate/n-hexane) to obtain the title compound 7 (13.7
mg, 59%).
[0591] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.71 (1H, m), 2.43
(1H, dd, J=13.0, 3.3 Hz), 2.35-1.90 (4H, m), 2.04 (3H, s),
1.74-1.19 (19H, m), 0.98 (3H, s), 0.94 (3H, d, J=6.4 Hz), 0.86 (3H,
d, J=7.0 Hz), 0.78 (6H, d, J=6.8 Hz), 0.67 (3H, s).
EXAMPLE 63
(1R,3aR,5aS,7S,9aS,11
aR)-7-Methoxymethoxy-1-[(S)-1-methoxypropan-2-yl]-9a,11a-dimethyl-1,2,3,3-
a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]chromen-5(10H)-one
(Compound 67)
[0592] Compound P55 (1.47 g) obtained in Reference Example 54 was
dissolved in acetic anhydride (53 mL), and sodium acetate (326 mg,
3.97 mmol) was added thereto, followed by stirring at 105.degree.
C. for 2 hours. The reaction mixture was left to cool, and then a
saturated aqueous ammonium chloride solution was added, followed by
extraction with ethyl acetate (50 mL.times.3). The organic layer
was washed with water and saturated brine, and dried over anhydrous
sodium sulfate. The residue obtained by concentration was purified
by silica gel column chromatography (5 to 20% ethyl
acetate/n-hexane) to obtain the title compound (582 mg, 54% (2
steps)).
[0593] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.69 (2H, m), 3.51
(1H, m), 3.38 (3H, s), 3.33 (1H, m), 3.32 (3H, s), 3.14 (1H, dd,
J=6.8 Hz, 15.8 Hz), 2.38-1.21 (18H, m), 1.05 (3H, d, J=6.6 Hz),
0.97 (3H, s), 0.69 (3H, s).
EXAMPLE 64
(1R,3aR,5aS,7S,9aS,11aR)-7-Hydroxy-1-[(S)-1-methoxypropan-2-yl]-9a,11a-dim-
ethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]chrome-
n-5(10H)-one (Compound 82)
[0594] Compound 67 (581.6 mg, 1.431 mmol) obtained in Example 63
was dissolved in tert-butanol (48 mL), and pyrimidium
p-toluenesulfonate (3.6 g; 14 mmol) was added thereto, followed by
stirring at 105.degree. C. for 4 hours. The reaction mixture was
cooled to 0.degree. C., a saturated aqueous ammonium chloride
solution was added, followed by extraction with ethyl acetate (50
mL.times.3). The organic layer was washed with saturated brine,
dried over anhydrous magnesium sulfate, and concentrated to yield a
residue. The residue was purified by silica gel column
chromatography, followed by purification by high-speed liquid
chromatography (60 to 100% acetonitrile/water) to obtain the title
compound (150 mg, 29%).
[0595] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.63 (1H, m),
3.35-3.30 (1H, m), 3.32 (3H, s), 3.14 (1H, dd, J=6.8 Hz, 16 Hz),
2.40-1.36 (19H, m), 1.05 (3H, d, J=6.4 Hz), 0.97 (3H, s), 0.69 (3H,
s).
EXAMPLE 65
(1R,3aR,5aS,7S,9aS,11aR)-1-[(S')-1-Isobutoxypropan-2-yl]-7-methoxymethoxy--
9a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopent-
a[h]chromen-5(10H)-one (Compound 65)
[0596] A crude product (2.34 g) of compound P63 obtained in
Reference Example 62 was treated with sodium acetate (520 mg, 6.34
mmol) in the same manner as Reference Example 46 to obtain the
title compound (1.10 g).
EXAMPLE 66
(1R,3aR,5aS,7S,9aS,11aR)-7-Hydroxy-1-[(S)-1-isobutyloxypropan-2-yl]-9a,11a-
-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]ch-
romen-5(10H)-one (Compound 81)
[0597] Compound 65 (1.10 g) obtained in Example 65 was treated with
pyrimidium p-toluenesulfonate (5.3 g, 21 mmol) in the same manner
as Example 63 to obtain the title compound (134 mg, 8.5%).
[0598] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.64 (1H, m), 3.37
(1H, dd, J=3.1 Hz, 9.0 Hz), 3.22-3.05 (3H, m), 2.42-1.25 (19H, m),
1.05 (3H, d, J=6.6 Hz), 0.97 (3H, s), 0.89 (6H, dd, J=2.4 Hz, 6.6
Hz), 0.69 (3H, s).
EXAMPLE 67
(1R,3
aR,5aS,7S,9aS,11aR)-7-Hydroxy-1-[(S)-1-hydroxypropan-2-yl]-9a,11a-di-
methyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]chrom-
en-5(10H)-one (Compound 95)
[0599] Compound P85 (126.8 mg, 0.2310 mmol) obtained in Reference
Example 66 was dissolved in THF (8 mL), and acetic acid (2.4 mL, 42
mmol) and 1.0 mol/L of a tetrabutylammoniumfluoride-THF solution
(10.5 mL, 10.5 mmol) were added thereto, followed by stirring at
room temperature for 6 days. A saturated aqueous sodium bicarbonate
solution which had been cooled to 0.degree. C. was added to the
reaction mixture, followed by extraction with ethyl acetate (30
mL.times.3). The organic layer was washed with saturated sodium
chloride, dried over anhydrous magnesium sulfate, and concentrated
to yield a residue. The residue was purified by silica gel column
chromatography (33% to 200% ethyl acetate/n-hexane) to obtain the
title compound (71 mg, 79%).
[0600] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.74-4.65 (2H, m),
3.65 (1H, dd, J=3.1 Hz, 10.5 Hz), 3.52 (1H, m), 3.42-3.38 (4H, m),
2.38-1.22 (19H, m), 1.07 (3H, d, J=6.6 Hz), 0.97 (3H, s), 0.70 (3H,
s).
EXAMPLE 68
(1R,3
aR,5aS,7S,9aS,11aR)-1-[(S)-1-Allyloxypropan-2-yl]-7-methoxymethoxy-9-
a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta-
[h]chromen-5(10H)-one (Compound 66)
[0601] Compound 95 (16.7 mg, 0.0425 mmol) obtained in Example 67
was dissolved in allylbromide (2 mL), and silver oxide (986 mg,
4.25 mmol) was added thereto, followed by stirring under light
shading at room temperature for 14 hours. Silver oxide was
separated by filtration, the filtrate was concentrated, and the
residue was purified by silica gel column chromatography (33% ethyl
acetate/n-hexane) to obtain the title compound (3.7 mg, 21%).
[0602] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.93 (1H, m),
5.32-5.15 (2H, m), 4.73-4.66 (2H, m), 3.96 (1H, d, J=5.7 Hz), 3.94
(1H, d, J=5.9 Hz), 3.50 (1H, m), 3.36 (3H, s), 3.35 (1H, m), 3.17
(1H, dd, J=7.3 Hz, 9.0 Hz), 2.30-1.13 (18H, m), 1.07 (3H, d, J=6.6
Hz), 0.97 (3H, s), 0.88 (3H, s).
EXAMPLE 69
(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-1-[(S)-1-propoxy-
propan-2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h-
]chromen-5(10H)-one (Compound 68)
[0603] Compound 66 (3.7 mg, 0.0086 mmol) obtained in Example 68 was
dissolved in ethyl acetate (2 mL), and 10% palladium-carbon (2.0
mg) was added thereto, followed by stirring at room temperature for
2 hours under a hydrogen atmosphere. The catalyst was separated by
filtration, followed by concentrating the filtrate to obtain the
title compound (2.8 mg, 76%).
[0604] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.66-4.60 (2H, m),
3.32 (1H, m), 3.20 (3H, s), 3.30-3.18 (3H, m), 3.02 (1H, dd, J=7.7
Hz, J=9.2 Hz), 2.32-1.15 (20H, m), 0.98 (3H, d, J=6.4 Hz), 0.87
(3H, s), 0.84 (3H, t, J=7.3 Hz), 0.62 (3H, s).
EXAMPLE 70
(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-1-[(S)-1-(2-meth-
ylallyloxyl)propan-2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]-
cyclopenta[h]chromen-5(10H)-one (Compound 96)
[0605] Compound 95 (7.6 mg, 0.019 mmol) obtained in Example 67 was
dissolved in allylbromide (0.5 mL), and silver oxide (986 mg, 4.25
mmol) was added thereto, followed by stirring under light shading
at room temperature for 12 hours. Silver oxide was separated by
filtration, the filtrate was concentrated, and then the residue was
purified by silica gel column chromatography (33% ethyl
acetate/n-hexane) to obtain the title compound (4.2 mg, 48%).
[0606] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 5.68 (1H, s), 5.10
(1H, s), 4.95-4.88 (2H, m), 4.75-4.65 (2H, m), 3.90-3.80 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 3.36-3.11 (2H, m), 2.37-1.25 (19H, m),
1.07 (3H, d, J=6.6 Hz), 0.97 (3H, s), 0.69 (3H, s).
EXAMPLE 71
(1R,3aR,5aS,7S,9aS,11aR)-1-[(2S)-1-(3-Hydroxy-2-methylpropoxy)propane-2-yl-
]-7-methoxymethoxy-9a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahyd-
robenzo[c]cyclopenta[h]chromen-5(10H)-one (Compound 70)
[0607] Compound P70 (13.6 mg, 0.0219 mmol) obtained in Reference
Example 73 was treated with 1.0 mol/L of a tetrabutylammonium
fluoride-THF solution and acetic acid in the same manner as
Reference Example 3 to obtain the title compound (8.6 mg, 84%).
[0608] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.38 (3H, s), 3.61-3.15 (7H, m), 2.81 (1H, m), 2.37-1.32 (19H, m),
1.04 (3H, d, J=6.6 Hz), 0.97 (3H, s), 0.86 (3H, dd, J=2.2 Hz, 7.0
Hz), 0.68 (3H, s).
EXAMPLE 72
(1R,3aR,5aS,7S,9aS,11aR)-1-[(2S)-1-(3-Methoxy-2-methylpropoxy)propan-2-yl]-
-7-methoxymethoxy-9a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydr-
obenzo[c]cyclopenta[h]chromen-5(10H)-one (Compound 71)
[0609] Compound 70 (15.1 mg, 0.0325 mmol) obtained in Example 71
was treated with silver oxide (1.50 g, 6.49 mmol) and methyl iodide
(3.0 mL) instead of allylbromide in the same manner as Example 67
to obtain the title compound (6.3 mg, 40%).
[0610] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 3.33 (3H, s), 3.37-3.08 (6H, m),
2.37-1.25 (19H, m), 1.04 (3H, d, J=6.4 Hz), 0.97 (3H, s), 0.94 (3H,
d, J=6.8 Hz), 0.69 (3H, s).
EXAMPLE 73
(R)-[(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-5-oxo-1,2,3-
,3a,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydrobenzo[c]cyclopenta[h]chromen-1-
-yl]octyl acetate (Compound 73)
[0611] Compound P78 (17.8 mg, 0.0340 mmol) obtained in Reference
Example 81 was dissolved in dichloromethane (3.4 mL), and
triethylamine (20.0 .mu.L, 0.143 mmol) and carbonyldiimidazole (8.3
mg, 0.051 mmol) were added thereto, followed by stirring at room
temperature for 11 hours. A saturated aqueous ammonium chloride
solution was added thereto, followed by extraction with ethyl
acetate (20 mL.times.3). The organic layer was washed with
saturated brine, dried over anhydrous magnesium sulfate, and
concentrated to yield a residue. The residue was purified by silica
gel column chromatography (33% ethyl acetate/n-hexane) to obtain
the title compound (7.0 mg, 41%).
[0612] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 2.37-0.85 (33H, m), 0.97 (3H, s), 0.93
(3H, d, J=6.6 Hz), 0.67 (3H, s).
EXAMPLE 74
(R)-[(1R,3aR,5aS,7S,9aS,11aR)-7-Hydroxy-9a,11a-dimethyl-5-oxo-1,2,3,3a,5,5-
a,6,7,8,9,9a,10,11,11a-tetradecahydrobenzo[c]cyclopenta[h]chromen-1-yl]oct-
yl acetate (Compound 97)
[0613] Compound 73 (7.0 mg, 0.014 mmol) obtained in Example 73 was
treated with pyrimidium p-toluenesulfonate (35 mg, 0.14 mmol) in
the same manner as Example 63 to obtain the title compound (1.9 mg,
30%).
[0614] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.05 (2H, t, J=6.8
Hz), 3.64 (1H, m), 2.42-1.12 (32H, m), 0.97 (3H, s), 0.93 (3H, d,
J=6.4 Hz), 0.67 (3H, s).
EXAMPLE 75
(1R,3aR,5aS,7S,9aS,11aR)-1-[(R)-8-Hydroxyoctan-2-yl]-7-methoxymethoxy-9a,1-
1a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]-
chromen-5(10H)-one (Compound 74)
[0615] Compound P81 (6.2 mg, 0.011 mmol) obtained in Reference
Example 84 was dissolved in THF (2 mL), water (0.5 mL) and
trifluoroacetic acid (0.5 mL), followed by stirring at 0.degree. C.
for 2 hours. Water was added to the reaction mixture, followed by
extraction with ethyl acetate (5 mL.times.3). The organic layer was
dried over anhydrous magnesium sulfate, and concentrated to yield a
residue. The residue was purified by silica gel column
chromatography (33% ethyl acetate/n-hexane) to obtain the title
compound (5.1 mg, 100%).
[0616] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.72-4.68 (2H, m),
3.64 (2H, t, J=6.6 Hz), 3.50 (1H, m), 3.38 (3H, s), 2.39-1.20 (31H,
m), 0.97 (3H, s), 0.93 (3H, d, J=6.6 Hz), 0.67 (3H, s).
EXAMPLE 76
(1R,3aR,5aS,7S,9aS,11aR)-1-[(R)-8-Azideoctan-2-yl]-7-methoxymethoxy-9a,11a-
-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]ch-
romen-5(10H)-one (Compound 98)
[0617] Compound 74 (7.3 mg, 0.016 mmol) obtained in Example 75 was
dissolved in toluene (2 mL), and triphenylphosphine (21 mg, 0.079
mmol), diphenylphosphoryl azide (17 .mu.L, 0.079 mmol) and diethyl
azodicarboxylate (17 .mu.L, 0.079 mmol) were added thereto,
followed by stirring at room temperature for 3 hours. Water was
added to the reaction mixture, followed by extraction with ethyl
acetate (5 mL.times.3). The organic layer was washed with saturated
brine, dried over anhydrous magnesium sulfate, and concentrated to
yield a residue. The residue was purified by silica gel column
chromatography (33% ethyl acetate/n-hexane) to obtain the title
compound (5.1 mg, 66%).
[0618] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.72-4.68 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 3.26 (2H, t, J=6.8 Hz), 2.37-1.20 (28H,
m), 0.97 (3H, s), 0.93 (3H, d, J=6.4 Hz), 0.67 (3H, s).
EXAMPLE 77
(R)-7-[(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-5-oxo-1,2-
,3,3a,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydrobenzo[c]cyclopenta[h]chromen-
-1-yl]octyl carbamate (Compound 75)
[0619] Compound 98 (5.1 mg, 0.011 mmol) obtained in Example 76 was
dissolved in ethyl acetate (3 mL), and di-tert-butyldicarbonate
(11.0 mg, 0.0525 mmol) and catalytic amount of 10% palladium-carbon
were added thereto, followed by stirring at room temperature for 2
hours under a hydrogen atmosphere. The catalyst was separated by
filtration, and the filtrate was concentrated to yield a residue.
The residue was purified by silica gel column chromatography (25%
ethyl acetate/n-hexane) to obtain the title compound (3.7 mg,
63%).
[0620] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.72-4.68 (2H, m),
4.49 (1H, br s), 3.52 (1H, m), 3.38 (3H, s), 3.15-3.05 (2H, m),
2.37-1.22 (37H, m), 0.97 (3H, s), 0.92 (3H, d, J=6.4 Hz), 0.67 (3H,
s).
EXAMPLE 78
(1R,3aR,3bR,5aS,9aR,9bR,11aR)-9a,11a-Dimethyl-1-[(R)-6-methylheptan-2-yl]d-
odecahydrobenzo[c]cyclopenta[h]chromen-5,7(3bH,8H)-dione (Compound
93)
[0621] Step 1; Commercially available 7-dehydrocholesterol (18.3 g,
47.6 mmol) was dissolved in dichloromethane (150 mL), and
N,N-diisopropylethylamine (24.9 mL, 143 mmol) and
chloromethylmethyl ether (7.23 mL, 95.2 mmol) were added thereto at
room temperature, followed by stirring overnight. A saturated
aqueous ammonium chloride solution was added to the reaction
mixture, followed by extraction with chloroform twice. The organic
layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by silica gel column
chromatography to obtain (S)-3-methoxymethoxy-5,7-cholestadiene
(18.1 g, 89%).
[0622] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.62 (s, 3H), 0.87
(d, J=7.0 Hz, 3H), 0.87 (d, J=7.0 Hz, 3H), 0.94 (d, J=6.4 Hz, 3HG),
0.94 (s, 3H), 0.95-2.03 (m, 22H), 2.09 (d, J=12.6 Hz, 1H), 2.32 (m,
1H), 2.52 (d, J=12.1 Hz, 1H), 3.38 (s, 3H), 3.50 (m, 1H), 4.71 (s,
2H), 5.38 (d, J=3.3 Hz, 1H), 5.57 (d, J=3.3 Hz, 1H).
[0623] Step 2; By using (S)-3-methoxymethoxy-5,7-cholestadiene
(18.1 g, 42.2 mmol) obtained in the Step 1,
(3S,5S,6S)-3-methoxymethoxy-7-cholesten-6-ol (13.9 g, 74%) was
obtained in the same manner as Reference Example 2.
[0624] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.54 (s, 3H), 0.85
(s, 3H), 0.87 (d, J=7.0 Hz, 3H), 0.87 (d, J=7.0 Hz, 3H), 0.92 (d,
J=6.4 Hz, 3H), 0.95-1.95 (m, 26H), 2.05 (m, 1H), 2.30 (m, 1H), 3.38
(s, 3H), 3.51 (m, 1H), 4.66-4.73 (m, 2H), 5.18 (s, 1H).
[0625] Step 3; By using
(3S,5S,6S)-3-methoxymethoxy-7-cholesten-6-ol (21.4 g, 47.9 mmol)
obtained in the Step 2, (3S,5S)-3-methoxymethoxy-7-cholesten-6-one
(20.1 g, 94%) was obtained in the same manner as Reference Example
4.
[0626] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.60 (s, 3H), 0.87
(d, J=7.0 Hz, 3H), 0.87 (s, 3H), 0.87 (d, J=7.0 Hz, 3H), 0.94 (d,
J=6.4 Hz, 3H), 0.95-2.37 (m, 26H), 3.38 (s, 3H), 3.53 (m, 1H), 4.69
(d, J=18.1 Hz, 1H), 4.71 (d, J=18.1 Hz, 1H), 5.73 (s, 1H).
[0627] Step 4; Sodium periodate (5.74 g, 26.8 mmol) and cerium
chloride heptahydrate (1.67 g, 4.48 mmol) were dissolved in water
(54 mL), ethyl acetate (108 mL), acetonitrile (108 mL) and
(3S,5S)-3-methoxymethoxy-7-cholestene-6-one (2.00 g, 4.48 mmol)
obtained in the Step 3 were added thereto and cooled to 0.degree.
C., and then ruthenium chloride hydrate (278 mg, 1.34 mmol) was
added thereto, followed by stirring for 1 hour. A saturated aqueous
sodium thiosulfate solution was added to the reaction mixture,
followed by extraction with ethyl acetate five times. The organic
layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated under reduced pressure to yield
a residue. The residue was purified by silica gel column
chromatography to obtain a crude product (1.70 g) of
(3S,5S,7S,8S)-3-methoxymethoxy-7,8-dihydroxycholestan-6-one. This
was dissolved in a mixed solvent of toluene (215 mL) and methanol
(21 mL), and lead tetraacetate (4.72 g, 10.7 mmol) was added
thereto at 0.degree. C., followed by stirring for 80 minutes. A
saturated aqueous sodium thiosulfate solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was washed with saturated brine, dried over
anhydrous magnesium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
column chromatography to obtain (1S,2R,5S)methyl
5-methoxymethoxy-2-methyl-2-[(1R,3aR,5R,7aR)-7a-methyl-1-[(R)-6-methylhep-
tan-2-yl]-4-oxooctahydro-1H-inden-5-yl]cyclohexanecarboxylate (937
mg, 44%).
[0628] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.59 (s, 3H), 0.85
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=5.7 Hz, 3H),
0.96-2.02 (m, 23H), 2.09-2.21 (m, 2H), 2.25-2.40 (m, 3H), 2.69 (dd,
J=3.6, 12.8 Hz, 1H), 3.36 (s, 3H), 3.45 (m, 1H), 3.64 (s, 3H), 4.66
(s, 2H).
[0629] Step 5; (1S,2R,5S)Methyl
5-methoxymethoxy-2-methyl-2-[(1R,3aR,5R,7aR)-7a-methyl-1-[(R)-6-methylhep-
tan-2-yl]-4-oxoocatahydro-1H-inden-5-yl]cyclohexanecarboxylate
(19.8 mg, 0.0414 mmol) obtained in the Step 4 was dissolved in
tert-butyl alcohol (0.414 mL), and pyrimidium p-toluenesulfonate
(104 mg, 0.414 mmol) was added thereto at room temperature,
followed by heating under reflux for 4 hours. The reaction mixture
was cooled to room temperature, and then a saturated aqueous sodium
bicarbonate solution was added, followed by extraction with ethyl
acetate three times. The organic layer was washed with saturated
brine, dried over anhydrous magnesium sulfate, and concentrated
under reduced pressure to yield a residue. The residue was purified
by preparative thin layer chromatography to obtain (1S,2R,5S)methyl
5-hydroxy-2-methyl-2-[(1R,3aR,5R,7aR)-7a-methyl-1-[(R)-6-methylheptan-2-y-
l]-4-oxooctahydro-1H-inden-5-yl]cyclohexanecarboxylate (7.2 mg,
40%).
[0630] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.59 (s, 3H), 0.85
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.93 (d, J=6.2 Hz, 3H),
0.98-2.31 (m, 26H), 2.37-2.50 (m, 2H), 2.86 (m, 1H), 3.57 (m, 1H),
3.66 (s, 3H).
[0631] Step 6; (1S,2R,5S)Methyl
5-hydroxy-2-methyl-2-[(1R,3aR,5R,7aR)-7a-methyl-1-[(R)-6-methylheptan-2-y-
l]-4-oxoocatahydro-1H-inden-5-yl]cyclohexanecarboxylate (365 mg,
0.840 mmol) obtained in the Step 5 was dissolved in toluene (8.0
mL), and 1.0 mol/L of a diisobutylaluminum hydride/toluene solution
(2.94 mL, 2.94 mmol) was added thereto at 0.degree. C. room
temperature, followed by stirring for 20 minutes. Methanol (1.0 mL)
was added to the reaction mixture, and sodium sulfate decahydrate
(4.74 g, 14.7 mmol) and chloroform (30 mL) were added thereto,
followed by stirring at room temperature for 8 hours. The reaction
mixture was filtered through a celite, and the filtrate was
concentrated under reduced pressure to yield a residue. The residue
was purified by silica gel column chromatography to obtain
(1R,3aR,3bR,5aS,7S,9aR,9bR,11aR)-9a,11a-dimethyl-1-[(R)-6-methy-
lheptan-2-yl]hexadecahydrobenzo[c]cyclopenta[h]chromen-5,7-diol
(146 mg, 43%).
[0632] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.82 (s, 3H), 0.85
(d, J=7.0 Hz, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.91 (d, J=6.9 Hz, 3H),
0.95-1.92 (m, 26H), 1.97-2.12 (m, 2H), 2.34 (m, 1H), 3.65 (m, 1H),
4.22 (m, 1H), 4.78 (m, 1H).
[0633] Step 7; By using
(1R,3aR,3bR,5aS,7S,9aR,9bR,11aR)-9a,11a-dimethyl-1-[(R)-6-methylheptan-2--
yl]hexadecahydrobenzo[c]cyclopenta[h]chromen-5,7-diol (54.5 mg,
0.134 mmol) obtained in the Step 6, compound 93 (42.5 mg, 79%) was
obtained in the same manner as Reference Example 4.
[0634] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.85 (d, J=7.0 Hz,
3H), 0.87 (s, 3H), 0.88 (d, J=7.0 Hz, 3H), 0.91 (d, J=6.9 Hz, 3H),
0.95-1.96 (m, 22H), 2.05-2.17 (m, 2H), 2.32-2.63 (m, 3H), 2.74 (dd,
J=12.6, 16.2 Hz, 1H), 3.00 (dd, J=5.0, 12.6 Hz, 1H), 4.59 (m,
1H).
EXAMPLE 79
(5S,8S,9S)-7-Oxa-8,9-epoxy-3,6-cholestadione (Compound 94)
[0635] Compound 1 (34.3 mg, 0.0856 mmol) obtained in Example 1 was
dissolved in dichloromethane (1.0 mL), and 3-chloroperbenzoic acid
(34.1 mg, 0.128 mmol) was added thereof at 0.degree. C., followed
by stirring for 90 minutes. A saturated aqueous sodium bicarbonate
solution was added to the reaction mixture, followed by extraction
with chloroform twice. The organic layer was dried over anhydrous
sodium sulfate, and concentrated under reduced pressure to yield a
residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (10.1 mg, 29%).
[0636] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.78 (s, 3H), 0.86
(d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.92 (d, J=6.6 Hz, 3H),
0.99-1.70 (m, 15H), 1.81-2.11 (m, 8H), 2.41-2.58 (m, 3H), 2.71
(ddd, J=1.5, 4.8, 16.1 Hz, 1H), 3.15 (dd, J=4.5, 13.8 Hz, 1H).
EXAMPLE 80
(1R,5aS,7S,9aR,9bR,11aR)-7-Hydroxy-9a,11a-dimethyl-1-[(R)-6-methylheptan-2-
-yl]-2,3,3,5a,6,7,8,9,9a,9b,10,11,11a-dodecahydrobenzo[c]cyclopenta[h]chro-
men-5(1H)-one (Compound 99)
[0637] Compound 56 (2.10 g, 4.63 mmol) obtained in Example 46 was
dissolved in tert-butanol (33 mL), and pyrimidium
p-toluenesulfonate (35.0 g; 140 mmol) was added thereto, followed
by stirring at 85.degree. C. for 2 hours. A saturated aqueous
sodium bicarbonate solution was added to the reaction mixture,
followed by extraction with ethyl acetate twice. The organic layer
was dried over anhydrous sodium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
column chromatography to obtain the title compound (26 mg,
1.4%).
[0638] .sup.1H NMR (CDCb) .delta.(ppm): 0.85 (s, 3H), 0.86 (d,
J=6.6 Hz, 3H), 0.87 (d, J=7.0 Hz, 3H), 0.93 (s, 3H), 0.94 (d, J=6.6
Hz, 3H), 0.99-1.80 (m, 18H), 1.81-2.05 (m, 3H), 2.15-2.30 (m, 3H),
2.36-2.58 (m, 2H), 3.63 (m, 1H).
EXAMPLE 81
(3S,5S)-7-Oxa-8-cholesten-6-on-3-ol isobutylate (Compound 100)
[0639] Compound 2 (60 mg, 0.15 mmol) obtained in Example 2 was
dissolved in dichloromethane (0.75 mL), and isobutyric acid
chloride (0.032 mL, 0.30 mmol) and pyridine (0.19 mL, 0.60 mmol)
were added thereto, followed by stirring at 0.degree. C. for 1
hour. A saturated aqueous sodium bicarbonate solution was added to
the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was dried over anhydrous sodium sulfate,
and concentrated under reduced pressure to yield a residue. The
residue was purified by preparative thin layer chromatography to
obtain the title compound (67 mg, 95%).
[0640] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H),
0.79-1.62 (m, 15H), 0.86 (d, J=6.6 Hz, 3H), 0.87 (d, J=6.6 Hz, 3H),
0.94 (d, J=6.5 Hz, 3H), 0.99 (s, 3H), 1.15 (d, J=6.9 Hz, 3H), 1.16
(d, J=7.1 Hz, 3H), 1.63-1.82 (m, 3H), 1.86-2.40 (m, 6H), 2.45 (dd,
J=13, 3.4 Hz, 1H), 2.56 (qq, J=6.6, 6.6 Hz, 1H), 4.71 (ddd, J=5.0,
5.0, 5.0 Hz, 1H).
EXAMPLE 82
(3S,5S)-7-Oxa-8-cholesten-6-on-3-ol furan-2-carboxylate (Compound
101)
[0641] Compound 2 (51 mg, 0.13 mmol) obtained in Example 2 was
dissolved in dichloromethane (0.65 mL), and furan-2-carboxylic acid
chloride (0.025 mL, 0.26 mmol) and pyridine (0.041 mL, 0.51 mmol)
were added thereto, followed by stirring at 0.degree. C. for 3
hours. A saturated aqueous sodium bicarbonate solution was added to
the reaction mixture, followed by extraction with ethyl acetate
twice. The organic layer was dried over anhydrous sodium sulfate,
and concentrated under reduced pressure. A residue obtained by
concentrating the organic layer under reduced pressure was purified
by preparative thin layer chromatography to obtain the title
compound (65 mg, 100%).
[0642] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H),
0.81-1.87 (m, 17H), 0.87 (d, J=6.4 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H),
0.94 (d, J=6.4 Hz, 3H), 1.03 (s, 3H), 1.91-2.55 (m, 8H), 5.00
(dddd, J=4.8, 4.7, 4.7, 4.6 Hz, 1H), 6.51 (dd, J=3.5, 1.8 Hz, 1H),
7.17 (dd, J=3.5, 0.7 Hz, 1H), 7.58 (dd, J=1.8, 0.7 Hz, 1H).
EXAMPLE 83
(S)-3-Methoxyimino-7-oxa-8-cholesten-6-one (Compound 102)
[0643] Compound 1 (40 mg, 0.10 mmol) obtained in Example 1 was
dissolved in ethanol (5 mL), and O-methylhydroxyamine hydrochloride
(25 mg, 0.30 mmol) and pyridine (0.024 mL, 0.03 mmol) were added
thereto, followed by stirring at room temperature for 3 hours. A
saturated aqueous ammonium chloride solution was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain the
title compound (39 mg, 93%) as an E/Z mixture.
[0644] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.69 (s, 1.5H), 0.69
(s, 1.5H), 0.80-1.62 (m, 16H), 0.87 (d, J=6.6 Hz, 1.5H), 0.87 (d,
J=6.6 Hz, 1.5H), 0.88 (d, J=6.6 Hz, 1.5H), 0.88 (d, J=6.6 Hz,
1.5H), 0.94 (d, J=6.3 Hz, 1.5H), 0.94 (d, J=6.3 Hz, 1.5H),
1.63-2.48 (m, 9.5H), 2.49 (dd, J=14, 4.3 Hz, 0.5H), 2.53 (dd, J=14,
3.6 Hz, 0.5H), 2.76 (ddd, J=14, 3.6, 1.3 Hz, 0.5H), 3.24 (ddd,
J=14, 4.9, 1.7 Hz, 0.5H), 3.62 (ddd, J=14, 4.3, 1.3 Hz, 0.5H), 3.81
(s, 1.5H), 3.82 (s, 1.5H).
EXAMPLE 84
(3R,5S)-3-Fluoro-7-oxa-8-cholesten-6-one (Compound 103) and
(5S)-7-oxa-3,8-cholestadien-6-one (Compound 104)
[0645] Compound 2 (80 mg, 0.20 mmol) obtained in Example 2 was
dissolved in dichloromethane (8 mL), and diethylaminosulfur
trifluoride (0.057 mL, 0.40 mmol) was added thereto, followed by
stirring at 0.degree. C. for 3 hours. Water was added to the
reaction mixture, followed by extraction with ethyl acetate twice.
The organic layer was dried over anhydrous sodium sulfate, and
concentrated under reduced pressure to yield a residue. The residue
was purified by preparative thin layer chromatography to obtain
compound 103 (44 mg, 54%) and compound 104 (29 mg, 37%).
[0646] Compound 103; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68
(s, 3H), 0.80-2.48 (m, 24H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.95 (s, 3H), 2.80 (dd, J=13, 3.7
Hz, 1H), 4.95 (br d, J=47 Hz, 1H).
[0647] Compound 104; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.69
(s, 3H), 0.76-1.67 (m, 12H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.93 (s, 3H), 0.94 (d, J=6.6 Hz, 3H), 1.74 (m, 1H),
1.86-2.53 (m, 9H), 2.59 (dd, J=11, 5.5 Hz, 1H), 5.61 (m, 1H), 5.73
(m, 1H).
EXAMPLE 85
(3S,5S)-3-Fluoro-7-oxa-8-cholesten-6-one (Compound 106)
[0648] Compound 105 (93 mg, 0.23 mmol) obtained in the Step 1 of
Example 10 was dissolved in dichloromethane (9 mL), and
diethylaminosulfur trifluoride (0.066 mL, 0.46 mmol) was added
thereto, followed by stirring at 0.degree. C. for 3 hours. Water
was added to the reaction mixture, followed by extraction with
ethyl acetate twice. The organic layer was dried over anhydrous
sodium sulfate, and concentrated under reduced pressure to yield a
residue. The residue was purified by preparative thin layer
chromatography to obtain the title compound (11 mg, 12%).
[0649] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68 (s, 3H),
0.80-2.54 (m, 25H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H),
0.93 (d, J=6.6 Hz, 3H), 1.00 (s, 3H), 4.95 (dtt, J=47, 11, 5.5 Hz,
1H).
EXAMPLE 86
(1R,5aS,7
S,9aS,11aR)-7-Hydroxy-9a,11a-dimethyl-1-[(R)-6-methylheptan-2-yl-
]-1,2,5a,6,7,8,9,9a,10,11,11a-decahydrobenzo[c]cyclopenta[h]chromen-5(10H)-
-one (Compound 107)
[0650] Compound 56 (2.9 g, 6.6 mmol) obtained in Example 46 was
dissolved in tert-butanol (47 mL), and pyrimidium
p-toluenesulfonate (50.0 g; 200 mmol) was added thereto, followed
by stirring at 80.degree. C. for 2 hours. A saturated aqueous
sodium bicarbonate solution was added to the reaction mixture,
followed by extraction with ethyl acetate twice. The organic layer
was dried over anhydrous sodium sulfate, and concentrated under
reduced pressure to yield a residue. The residue was purified by
column chromatography to obtain the title compound (3.6 mg,
0.14%).
[0651] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.84-1.69 (m, 12H),
0.86 (s, 3H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H), 0.94
(d, J=6.6 Hz, 3H), 1.03 (s, 3H), 1.76 (m, 1H), 1.87-2.22 (m, 3H),
2.23-2.87 (m, 7H), 3.66 (m, 1H), 5.76 (br s, 1H).
EXAMPLE 87
(3S,5R)-3-Hydroxy-3-methyl-7-oxa-8-cholesten-6-one (Compound 108)
and (3S,5S)-3-hydroxy-3-methyl-7-oxa-8-cholesten-6-one (Compound
109)
[0652] Compound 1 (73 mg, 0.18 mmol) obtained in Example 1 was
dissolved in THF (6.0 mL), a 3.0 mol/L THF solution of
methylmagnesium chloride (0.12 mL, 0.37 mmol) was added thereto at
-78.degree. C., followed by stirring at the same temperature for
2.5 hours. Water was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was dried
over anhydrous sodium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by
preparative thin layer chromatography to obtain compound 108 (36
mg, 41%) and compound 109 (43 mg, 49%).
[0653] Compound 108; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68
(s, 3H), 0.81-1.87 (m, 19H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.92 (s, 3H), 0.93 (d, J=6.6 Hz, 3H), 1.31 (s, 3H),
1.89-2.43 (m, 6H), 2.87 (dd, J=13, 3.6 Hz, 1H).
[0654] Compound 109; .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.68
(s, 3H), 0.80-1.82 (m, 19H), 0.87 (d, J=6.6 Hz, 3H), 0.88 (d, J=6.6
Hz, 3H), 0.94 (d, J=6.6 Hz, 3H), 0.97 (s, 3H), 1.26 (s, 3H),
1.89-2.42 (m, 6H), 2.43 (dd, J=13, 3.3 Hz, 1H).
EXAMPLE 88
(1R,3aS,5
aS,7S,9aS,11aR)-7-Hydroxy-9a,11a-dimethyl-1-[(R)-6-methylheptan--
2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]chrome-
n-5(10H)-one (Compound 110)
[0655] Compound 56 (110 mg, 0.250 mmol) in obtained in Example 46
was dissolved in tert-butanol (5 mL), and pyrimidium
p-toluenesulfonate (1.9 g; 7.4 mmol) was added thereto, followed by
stirring at 80.degree. C. for 1 hour. A saturated aqueous sodium
bicarbonate solution was added to the reaction mixture, followed by
extraction with ethyl acetate twice. The organic layer was dried
over anhydrous sodium sulfate, and concentrated under reduced
pressure to yield a residue. The residue was purified by column
chromatography to obtain the title compound (84 mg, 85%).
[0656] .sup.1H NMR (CDCl.sub.3) .delta.(ppm): 0.75-1.60 (m, 15H),
0.83 (d, J=6.6 Hz, 3H), 0.84 (d, J=6.6 Hz, 3H), 0.89 (s, 3H), 0.90
(d, J=6.6 Hz, 3H), 0.92 (s, 3H), 1.62-2.20 (m, 9H), 2.21-2.38 (m,
2H), 3.62 (m, 1H).
EXAMPLE 89
(S)-2-[(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-5-oxo-1,2-
,3a,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydrobenzo[c]cyclopenta[h]chromen-1-
-yl]propanal (Compound 111)
[0657] Compound 95 (20 mg, 0.051 mmol) obtained in Example 67 was
dissolved in dichloromethane (3 mL), and
1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (Dess-Martin
Periodinane) (65 mg, 0.15 mmol) was added thereto, followed by
stirring at room temperature for 40 minutes. A saturated aqueous
sodium bicarbonate solution was added to the reaction mixture,
followed by extraction with chloroform (20 mL.times.3). The organic
layer was washed with saturated brine, dried over anhydrous
magnesium sulfate, and concentrated to yield a residue, which was
purified by silica gel column chromatography (25% to 50% ethyl
acetate/n-hexane) to obtain the title compound (11 mg, 53%).
[0658] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 9.61 (1H, d, J=2.9
Hz), 4.75-4.65 (2H, m), 3.53 (1H, m), 3.39 (3H, s), 2.45-1.22 (18H,
m), 1.15 (3H, d, J=6.8 Hz), 0.98 (3H, s), 0.73 (3H, s).
EXAMPLE 90
(1R,3aR,5aS,7S,9aS,11aR)-1-[(S)-1-Isobutylaminopropan-2-yl]-7-methoxymetho-
xy-9a,11a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclop-
enta[h]chromen-5(10H)-one (Compound 69)
[0659] Compound 111 (23 mg, 0.059 mmol) obtained in Example 89 was
dissolved in THF (3 mL), and isobutyl amine (30 .mu.L, 0.30 mmol)
and sodium triacetoxyborohydride (62 mg, 0.30 mmol) were added
thereto, followed by stirring at room temperature for 1 hour. Water
was added to the reaction mixture, followed by extraction with
ethyl acetate (10 mL.times.2). The organic layer was dried over
anhydrous magnesium sulfate, and concentrated to yield a residue.
The residue was purified by silica gel column chromatography (50%
ethyl acetate/n-hexane) to obtain the title compound (6.0 mg,
22%).
[0660] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.52 (1H, m), 3.38 (3H, s), 2.77 (1H, m), 2.57-1.25 (23H, m), 1.10
(3H, d, J=6.4 Hz), 0.99-0.96 (9H, m), 0.70 (3H, s).
EXAMPLE 91
(1R,3aR,5aS,7S,9aS,11aR)-7-Hydroxy-1-[(S)-1-isobutylaminopropan-2-yl]-9a,1-
1a-dimethyl-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopenta[h]-
chromen-5(10H)-one (Compound 83)
[0661] By using compound 69 obtained in Example 90, the title
compound was obtained in the same manner as Example 48.
[0662] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 3.65 (1H, m), 2.78
(1H, m), 2.60-1.25 (24H, m), 1.10 (3H, d, J=6.5 Hz), 0.98-0.96 (9H,
m), 0.69 (3H, s).
EXAMPLE 92
(1R,3aR,5aS,7S,9aS,11aR)-7-Methoxymethoxy-9a,11a-dimethyl-1-[(S)-1-morphol-
inopropan-2-yl]-1,2,3,3a,5a,6,7,8,9,9a,11,11a-dodecahydrobenzo[c]cyclopent-
a[h]chromen-5(10H)-one (Compound 72)
[0663] By using morpholine and compound 111 obtained in Example 89,
the title compound was obtained in the same manner as Example
90.
[0664] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.75-4.65 (2H, m),
3.75-3.60 (4H, m), 3.53 (1H, m), 3.39 (3H, s), 2.49-1.15 (24H, m),
1.03 (3H, d, J=6.4 Hz), 0.98 (3H, s), 0.70 (3H, s).
EXAMPLE 93
7-Oxa-ergosta-8(9),24(28)-dien-3,6-dione (Compound 112)
[0665] 7-Oxa-ergosta-8(9),24(28)-dien-6-on-3-ol (10.6 mg, 0.0256
mmol) obtained in Reference Example 85 was dissolved in
dichloromethane (2 mL), followed by cooling to 0.degree. C., and
Dess-Martin Periodinane (48.0 mg, 0.113 mmol) was added thereto,
followed by stirring for 3 hours. An aqueous solution of sodium
bicarbonate was added to the reaction mixture and extracted with
chloroform (5 mL.times.3), followed by purification by silica gel
column chromatography (33% ethyl acetate/n-hexane) to obtain the
title compound (5.5 mg, 52%).
[0666] .sup.1H NMR .delta.(ppm, CDCl.sub.3): 4.73 (1H, br s), 4.66
(1H, br s), 2.81-2.73 (2H, m), 2.62-1.26 (21H, m), 1.17 (3H, s),
1.03 (6H, dd, J=1.8, 6.8 Hz), 0.98 (3H, d, J=6.6 Hz), 0.72 (3H,
s).
EXAMPLE 94
Proliferation Promoting Agent for Neural Stem Cells
[0667] A proliferation promoting agent for neural stem cells
containing Compound 1 is obtained by preparing a DMSO solution of
Compound 1 (0.1 mmol/L) according to the conventional method.
INDUSTRIAL APPLICABILITY
[0668] The present invention can be used for a promoting
proliferation of neural stem cells for the purpose of treatment of
disease such as Parkinson's disease, Alzheimer's disease, Down
syndrome, cerebrovascular disorders, stroke, spinal cord injury,
triplet repeat disease, multiple sclerosis, amyotrophic lateral
sclerosis, polyneuropathy, epilepsy, anxiety disorders,
schizophrenia, depression or manic depressive psychosis.
* * * * *