Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
CA 02348319 2001-05-24
TITLE OF THE INVENTION:
CYCLOHEXENOL DERIVATIVE AND MEDICAMENT CONTAINING THE SAME
BACKGROUNL) OF THE I:N'VENTION
Field of t.he Invention:
The present invention relates to novel cyclohexenol
derivatives which have excellent neurocyte growth-
accelerating effects and are useful as agents for
preventing or treating diseases caused by nervous
degeneration, and medicaments comprising such a compound.
Description of the Background Art:
Diseases causeci by nervous degeneration are diseases
in which neurocytes are gradually lost in a central
nervous system, and include diseases the main locus of
which is situated at the cerebral cortex, such as
Alzheimer's dementia and Pick disease, diseases the main
locus of which is situated at the cerebral basal ganglia,
such as Parkinson's disease and Huntington's disease,
diseases the main locus of which is situated at the
cerebellum, such as spino-cerebellar degeneration, and
diseases the main locus of which is situated at the spinal
cord, such as amyotrophic lateral sclerosis. Of these,
amyotrophic lateral sclerosis (ALS) is a lethal nervous
disease characterized by selective disorders of motor
neurons in the cortex, brain stem and spinal cord, and
principal symptoms thereof include progressive muscular
atrophy, acceleration of deep tendon reflex, etc. With
1
CA 02348319 2001-05-24
respect to parts of:. familial ALS and sporadic ALS, point
mutations of a Cu/2n superoxide dismutase (SOD) gene have
been receritly reported to be causative genes and have
attracted attentiori. (Deng, H. et al., Science, 261, 1047-
1051, 1993; Rosen, DR et al., Nature, 363, 59-62, 1993;
and Jones, CT. et al., Lancet, 342, 1050-1061, 1993).
On the other hand, nerve growth factors (NGF) and
brain derived neurotrophic factors (BDNF)are said to cause
the secretion of proteins which regulate the long-term
survival and mutation of nerves in the central nervous
system and the peripheral nervous system, and to take
charge of memory, learning and the like in mammals. In
recent years, it has been reported that such factors are
useful for the prevention or treatment of diseases caused
by nervous degeneration, including Alzheimer's dementia
and Parkinson's disease.
Since such factors are proteins having a high
molecular weight, however, they cannot pass through a
blood-brain barrier, and so the use thereof is not
realistic as a method for treating humans. The use of
nerve-protecting agents (antioxidants and antistimulants),
neuroregeneratives and neurotrophic factors has also been
attempted for the treatment for ALS. However, only very
weak efficacies have been reported, and therefore they are
not satisfactory for the above treatment.
SUMMARY OF' THE INVENTION
2
CA 02348319 2001-05-24
It is an object of the present invention to provide
an agent which compr:ises a compound of a low molecular
weight capable of passing through the blood-brain barrier,
having a nerve growth factor-like effect and which is
useful for the prevention or treatment of various diseases
caused by nerve degeneration.
With the foregoing in mind, the present inventors
have carried out an extensive investigation. As a result,
it has been found that specific cyclohexenol derivatives
have excellent neurocyte growth-accelerating effects
including neurite-outgrowing ability, and are useful for
the prevention or treatment of diseases caused by nerve
degeneration, thus leading to completion of the present
invention.
According to the present invention, there is thus
provided a cyclohexeriol derivative represented by the
following general formula (1):
OH
R5
R I (1)
1
R2 4 X-OH
R3
wherein R1 to R5 are independently a methyl group or a
hydrogen atom, and X is an alkylene or alkenylene group
having 10 to 28 carbon atoms.
According to the present invention, there is also
3
CA 02348319 2001-05-24
provided a medicamen;~ comprising the cyclohexenol
derivative as an act:ive ingredient.
According to the present invention, there is further
provided a neurocyte growth-accelerating agent comprising
the cyclohexenol derivative as an active ingredient.
According to the present invention, there is further
provided an agent foi.^ preventing or treating a disease
caused by nervous deqeneration, comprising the
cyclohexenol derivative as an active ingredient.
According to the present invention, there is still
further provided a pharmaceutical composition comprising
the cyclohexenol derIvative and a pharmaceutically
acceptable carrier.
According to the present invention, there is also
provided use of the cyclohexenol derivative for the
manufacture of a medicament.
According to the present invention, there are also
provided a method for accelerating the growth of
neurocytes and further provided a method for treating a
disease caused by nervous degeneration, which are
characterized by administering the cyclohexenol derivative
in an effective amount.
The cyclohexenol derivatives (1) according to the
present invent.ion exhibit neurocyte growth-accelerating
effects such as neurocyte-growing effects and neurite-
outgrowing effects arid are hence useful as agents for
preventing or treatiriq diseases caused by nervous
4
CA 02348319 2001-05-24
degeneration, such as Alzheimer's dementia, Pick disease,
Parkinson's disease, Huntington's disease, spino-
cerebellar degeneration and amyotrophic lateral sclerosis.
The above and other objects, features and advantages
of the present invenwion will be readily appreciated as
the same becomes bet-~~er understood from the preferred
embodiments of the present invention, which will be
described subsequent:Ly in detail, taken in conjunction
with the accompanying drawings, and from the appended
claims.
BRIEF DESCRIPTION OF THE DRAWINGS
Fig. 1 diagrammatically illustrates the effect of
Compound 1(10-9 to 10-5 M) on neurocyte growth, determined
by paired one-way analysis of variance (ANOVA), followed
by Newman-Keuls multiple comparison test.
Fig. 2 diagramrnatically illustrates the effect of
Compound 1(10-9 to 10-,5 M) on neurite outgrowth,
determined by paired one-way arialysis of variance (ANOVA),
followed by Newman--Keul.s multiple comparison test.
DETAILED DESCRIPTION OF' THE PREFERRED EMBODIMENTS
In the general f`ormula (1), X is ari alkylene or
alkenylene group having 10 to 28 carbon atoms. The
alkylene or alkenylene group referred to in the present
invention may be either linear or branched. Of such
groups X, a linear alkylene group having 10 to 28 carbon
5
CA 02348319 2001-05-24
atoms is niore preferred, with a linear alkylene group
having 10 to 18 carbon atoms being particularly preferred.
R1 to R5 are i.ndependently a hydrogen atom or a
methyl group, and at least one of them is preferably a
methyl group.
The compounds represented by the general formula (1)
may be in the form of a pharmaceutically acceptable salt,
or a solvate or hydrate thereof. Examples of preferable
salts include salts with alkali metals such as sodium,
potassium and lithium, and salts with alkaline earth
metals suc:h as magnesium and calcium.
Various isomers may exist, i.n the compounds (1), and
these isonters may be included in the present invention.
The cyclohexenol derivatives (1) according to the
present invention can be prepared, for example, in
accordance with the following Process 1:
[Process 1.]
25
6
CA 02348319 2001-05-24
0 0
R5 R5
Bromination R I Reduction
1
RR2 0 R2 7 s a Br
Rs R4 R R
(A) (B)
OH OZ
R5 Protect:ion of R5 1) tert-BuLi
2)gr-X1-C ¾
I hydroxy~l, grouP I _
1 R1
R2 Br
i7 R3 Ra Br R3 Ra
(C) (D)
OZ OH
R5 1)Mg R5
2)Br-X2-0Z'
RI 3) Deprotection R1
R2 X1-0~ R2 3 a X-OH
Rs Ra R R
CE) <1)
wherein R' to R5 are independently a hydrogen atom or a
methyl group, Z and Z' are independently a protecting
group for the hydroxyl group, X1 is an alkylene or
alkenylene group having 5 to 10 carbon atoms, X2 is an
alkylene or alkenylene group having 4 to 18 carbon atoms,
and X has the same meaning as defined above.
More specifically, cyclohexanedione [compound (A)]
is brominated into a bromide [compound (B)]. This
7
CA 02348319 2001-05-24
compound is reduced with a hydride into a cyclohexenol
derivative [compound (C)]. After the hydroxyl group in
the compound (C) is then protected to form a compound (D),
the protected compound is chloroalkylated or
chloroalkenylated into a halide [compound (E)]. After the
resultant compound is reacted with metal magnesium, a
halogenoalkanol the hydroxyl group of which has been
protected is allowed tc act on the reaction product to
alkylate cr alkenylate the reaction product. The
protecting group is then separated, whereby a cyclohexenol
derivative (1) acccrding to the present invention can be
obtained.
As a brominating agent in the bromination of the
compound (A), there is used (a) carbon tetrabromide, (b)
N-bromoimide type brominating agent such as N-bromo-
succinimid.e, N-brom.oglutarimide or N,N',N"-tribromo-
isocyanuric acid, cr (c) a bromopyridinium perbromide such
as dimethylaminopyridinium bromide perbromide or
pyridinium bromide perbromide. These reagents are
generally used in an amount of 1 to 10 equivalents,
preferably 1 to 2 equiva.lents. The bromination reaction
is generally performed at 0 to 5 C for 1 to 2 hours. It
is desirable to use an. ordinary inert solvent as a solvent
for the reaction. Examples of usable solvents include
halogenated hydrocarbons, hydrocarbens, ethers and
dimethylformamide.
In the reduction of the compound (B) into the
8
CA 02348319 2001-05-24
compound (C), there is preferred a reducing agent
generally used in the reduction of a carbonyl group, for
example, tetramethylammonium borohydride,
tetramethylammoniurn. borohydride triacetate, lithium
borohydricle, sodiuni. boro- hydri.de, sodium
trimethoxyborohydride, sodium triacetoxy- borohydride,
diisobutylaluminum hydride or the like. The amount of
such a reclucing agent used varies with reaction time, but
is generally within a range of 1 to 10 equivalents,
preferably 1 to 2 equivalents. As a solvent for the
reaction used, tolue;:-ie, acetonitrile, ethanol or
tetrahydrofuran is used. The reaction is generally
performed at 0 to 5 C:: for 1 to 2 hours.
As the protecting group used for the protection of
the hydroxyl group in the compound (C), there is preferred
an alkoxyalkyl group such as ari 1-ethoxyethyl,
tetrahydropyranyl cr tetrahydrofuranyl group. The
introduction of such a protecting group for the hydroxyl
group is conducted by reacting the cyclohexenol derivative
(C) with its corresponding vinyl ether under weakly acidic
conditions. Specific examples of the vinyl ether used
include ethyl vinyl ether, 2,3-dihydropyran and 2,3-
dihydrofuran.
The halogenoalkylation or halogenoalkenylation of
the compound (D) is preferably performed under conditions
of a low temperature iri the presence of a metallic
compound such as tert-butyllithium in a solvent such as
9
CA 02348319 2001-05-24
tetrahydrofuran.
The reaction of the compound (E) with metal
magnesium is carried out in a solvent such as diethyl
ether, tetrahydrofuran or dioxane, and the reaction of the
reaction product thereof with the halogenoalkanol can be
performed under ordirlary conditions used for a Grignard
reaction.
The removal of the hydroxyl protecting group is
performed under acid:_c conditions in an alcohol solvent
such as methanol, ethanol or butanol or a ketone solvent
such as acetone or methyl ethyl ketone. As an acid, there
is used an inorganic acid such as phosphoric acid or
hydrochloric acid; arl organic acid such as p-
toluenesulfonic acid; or an organic carboxylic acid such
as acetic acid.
The compounds (1) according to the present invention
can also be obtained by reducing a compound (F) (Process
2) or oxidizing a compound (G) (Process 3), said compounds
being obtained in accordance with the respective processes
described in WO 99/08987.
[Process 2]
0 OH
R5 R5
Reduction I
R~ I Rl X-
R2 OH
R2 R3 Rq X-GH R3 Rq
(F) (1)
CA 02348319 2001-05-24
[Process 3]
OH
R5 1) Oxidation R5
2) Deprotection
1
Rl R
RR2 X-OAr R2 X-OH
R3 R4 R3 R4
(G) (1)
wherein R' to R5 and X have the same meanings as defined
above.
The thus-obtairied cyclohexenol derivatives (1)
according to the present invention exhibit excellent
neurocyte growth-accelerating effects such as neurocyte-
growing effect and neurite-outgrowing effect, and pass
through a blood-brairi barrier because they are low-
molecular. Accordingly, they are useful as neurocyte
growth-accelerating agents and in its turn as agents for
preventing or treating mammalian diseases caused by
nervous degeneration (including such human diseses), such
as amyotrophic lateral sclerosis.
The medicaments according to the present invention
can be administered by either oral administration or
parenteral administration as injections (intramusclar,
subcutaneous and inti.-avenous), suppositories and the like.
In the formulation of an oral preparation, an
excipient, and optiorially a binder, a disintegrator, a
lubricant, a colorant, a taste and smell corrigent and/or
the like are added to the compound according to the
11
CA 02348319 2001-05-24
present invention, and the resultant mixture is then
prepared into tablets, coated tablets, granules, capsules,
solutions, syrups, elixirs, oil. or aqueous suspensions,
etc. in accordance with a method known per se in the art.
Examples of the excipient include lactose, corn
starch, sucrose, glucose, sorbit and crystalline cellulose.
Examples of the binder include polyvinyl alcohol,
polyvinyl ether, ethyl cellulose, methyl cellulose, gum
arabic, tragacanth gum, gelatin, shellac, hydroxypropyl
cellulose, hydroxypropyl starch and polyvinyl pyrrolidone.
Examples of the disintegrator include starch, agar,
gelatin powder, crys1:_all.ine cellulose, calcium carbonate,
sodium hydrogencarbonate, calcium citrate, dextran and
pectine. Examples of the lubricant include magnesium
stearate, talc, polyethylene glycol, silica and hardened
vegetable oils.
As the colorant, there may be used any colorant
allowed to add to medicinal compositions. As the taste
and smell corrigent., there may be used cocoa powder,
menthol, aromatic acids, peppermint oil, borneol, cinnamon
powder or the like. 7he tablets and granules may be
suitably c:oated with sugar, gelatin or the like as needed.
In the case where an injection is prepared, a pH
adjustor, a buffer, a stabilizer, a preservative and/or
the like are added to prepare the resultant mixture into a
subcutaneous injection, intramuscular injection or an
intravenous injecti.o:i in accordarice with a method known
12
CA 02348319 2001-05-24
per se in the art.
The injection rnay also be provided as a solid form
for preparing upon use by contairiing a solution into a
container and then drying the solution by lyophilization
or the like. A dose or doses of the injection may also be
contained in a container.
With respect to the dose of the cyclohexenol
derivative (1) according to the present invention as a
medicament, it is genera.lly administered in a dose ranging
from 0.01 to 1,000 mg, preferably from 0.1 to 100 mg, per
day for ar.. adult. Meanwhile, the administration may be
once per day, or may be divided into 2 to 4 times per day.
The present invention will hereinafter be described
in detain by the following Examples.
Referential Example 1:
(1) Synthesis of 3.-br_omo-5,5-dimethyl-2-cyclohexenone:
A sclution witli carbon tetrabromide (9.94 g)
dissolved in 20 ml of dimethylformamide was added to a
well-stirred solution of copper bromide (54 mg, 0.4 mmol),
1,10-phenanthroline monohydrate (148 mg, 0.8 mmol), copper
powder (1.42 g, 22.4 mmol), ir.orl powder (2.09 g, 37.5
mmol) and 5,5-dimet.hyl.cyclohexane-1,3-dione (2.11 g, 15.0
mmol) in 40 ml of dimet.hylformami.de under an argon stream.
The temperature was controlled by an ice/water bath.
After 1.5 hours, the reaction mi_~:ture was hydrolyzed with
1N hydrochloric acid. The aqueous layer was extracted 3
times with diethyl ether. The combined organic layers
13
CA 02348319 2001-05-24
were washed with a saturated solution of ammonium chloride
and a saturated saline, dried over magnesium sulfate and
then concentrated. The resultant reaction product was
isolated by flash c:hromatography (hexane/ethyl acetate;
9/1) to give 2.8 g(y:ield: 94%) of 3-bromo-5,5-dimethyl-2-
cyclohexenone as a yellow oil.
Mass: 199 (M+).
TLC: (hexane/ethyl acetate; 9/1) Rf = 0.35.
IR (KBr) cm-1: 2960; 1688; 1.339; 1269; 1220.
UV: a,max = 245 nm (E = 5000) .
1H-NMR (200 MHz; CD(:'-'13) ; S (ppm)
1. 07 (s, 6H, -CH3) ;:? . 24 ( s, 2H, H-6) ; 2. 68 (d, 9J=1 . 8Hz, 2H, H-4 );
6.45-6.47(t,'J=1.8Hz,2H,H-2).
13C-NMR (SC) MHz; CDC13) ; S (ppm)
28. 1(C-7, 8) ; 34.8 ((;--5) ; 50.3 (C-4*) ; 50.4 (C-6*) ;
131.5(C-2); 148.3((::--3).
(2) Synthesis of 3-bromo-5,5-dimethyl-2-cyclohexenol:
To a solution with 3-bromo-5,5-dimethyl-2-
cyclohexenone (0.5g, 2.6 mmol) dissolved in 60 ml of
toluene was added diisobutylaluminum hydride (1 M toluene
solution, 4 ml, 4 mmol) at 0 C under an argon stream.
After 2 hours, the reaction mixture was hydrolyzed by
adding methanol, purified water and 1N hydrochloric acid.
The aqueous layer was extracted 3 times with diethyl ether.
The combined organic layers were washed with a saturated
saline, dried over magnesium sulf:ate and then concentrated.
The residue was purified by flash chromatography
14
CA 02348319 2001-05-24
(hexane/ethyl acetate; 7/3) to give 504 mg (yield: 98%) of
3-bromo-5,5-dimethyl-2-cyclohexenol.
Mass: 201 (M+).
TLC: (hexane/ethyl acetate; 7/3) Rf = 0.36.
'H-NMR (200 MHz; CDCl;,) ; 8(ppm) :
0. 97 (s, A, -CH3) , 1. 03 (s, 3H, -CH3) ; 1. 51-1. 61 (m, 1H, H-6eq) ;
1.76-1.89(m,1H,H-6ax); 2.08-2.22(m,1H,H-4eq);
2.29-2.43(m,1H,H-4aq); 4.20-4.35(m,1H,H-1);
6. 08-6. 13 (m, 1H, H--2 ).
(3) Synthesis of 1-k:>romo-5,5-dimethyl-3-tetrahydro-
pyranyloxycyclohexene:
To a solution with 3-bromo-5,5-dimethyl-2-
cyclohexenol (0.97g, 4.8 mmol) dissolved in 20 ml of
methylene chloride were added pyridine p--toluenesulfonate
(200 mg, 0.8 mmol) and 3,4-dihydro-2H-pyran (1.9 ml, 19
mmol). After 12 hours, the reaction mixture was
neutralized with a saturated solution of sodium hydrogen-
carbonate and extracted 3 times with diethyl ether. The
combined organic layers were washed with a saturated
saline, dried over magnesium sulfate and then concentrated.
The residue was purified by column chromatography
(hexane/ethyl acetate; 100/0 to 90/10) to give 1.38 g
(yield: 94%) of 1-bromo-5,5-dimethyl-3-tetrahydro-
pyranyloxvcycl ohexE:>ne .
Mass: 289 (M+).
TLC: (hexane/ethyl acetate; 7/3) Rf = 0.6.
1H-NMR (2C0 MHz; CDC13); S (ppm):
CA 02348319 2001-05-24
0.96(s,3H,-CH3), 1.01(s,3H,-CH3); 1.29-1.48(m,1H,H-6eq);
1. 49-1 . 62 (m, 4H, H-3' , 4' ); 1. 69-1.. 78 (m, 4H, H-2' , 6ax) ;
2.07-2.20(m,1H,H-4eq); 2.30-2.43(m,1H,H-4aq);
3.45-3.58(m,1H,H-5'eq); 3.83-3.97(m,1H,H-5'ax);
4.18-4.33(m,1H,H-1); 4.70-4.78(m,1H,H-1');
6.08-6.21(m,1H,H-2).
13C-NMR (50 MHz; CDC13) ; S (ppm)
19.6(C-3'); 25.5(C-4'); 26.2(C-7*); 26.5(C-8*);
30.6-31.1(C-4); 33.0(C-5); 40.5-41.9(C-2*); 48.9(C-6);
62.5-62.7(C-5'); 71.9-72.3(C-3); 97.6(C-1'); 124.1(C-1);
128.3-130.0(C-2).
(4) Synthesis of 1-(6-chlorohexyl)-5,5-dimethyl-3-
tetrahydropyranyloxycyclohexene:
To a solution with 1-bromo-5,5-dimethyl-3-
tetrahydropyranyloxycyclohexene (0.54g, 1.9 mmol)
dissolved in 5 ml of tetrahydrofuran was added dropwise
tert-butyl.lithium (1.7 M in pentane, 3 ml, 5.1 mmol) at
-78 C under an argon stream. After 30 minutes, 1-bromo-6-
chlorocyclohexane (0.4 ml, 2.7 mmol) was added. After a
reaction was conducted for 1 hour, the reaction mixture
was hydrolyzed by purified water at 0 C. The aqueous
layer was extracted 3 times with diethyl ether. The
combined organic layers were washed with a saturated
saline, dried over magnesium sulfate and then concentrated.
The residue was purified by flash chromatography
(hexane/ethyl acetate; 95/5) to give 345 mg (yield: 60%)
of 1-(6-chlorohexyl.)-5,5-dimethyl-3-tetrahydropyranyloxy-
16
CA 02348319 2001-05-24
cyclohexene.
Mass: 329 (M+)
TLC: (hexane/ethyl acetate; 95/5) Rf = 0.35.
IR (KBr) c:m 1: 3027; 2949; 1360; 1022.
UV: kmax = 204 nm (E = 1000) .
1H-NMR (200 MHz; CDC13) ; 6 (ppm)
0. 88 (s, 3H, -CH3) ; 0. 98 (s, 3H, -CH3) ,
1. 21-1 . 37 (m, 4H, H-3' , 4' ); 1. 38-1. . 50 (m, 4H, H-3", 4") ;
1.52-1.69(m,6H,H-4,2',2"); 1.71-1.85(m,2H,H-5');
1.90-2.05(m,4H,H-1',6); 3.48-3.57(m,3H,H-5',6');
3.88-4.03(m,1H,H-5'); 4.18-4.32(m,1H,H-3);
4. 73-4 . 7 9(m, 1H, H-1 "); 5. 42-5 . 47 (m, 1H, H-2 ).
13C-NMR (50 MHz; CD(:;13) ; 8 (ppm) :
19.9(C-3"); 25.6(C-3"); 26.3(C-6'); 26.8(C-7*);
27.2(C-8*); 28.3(C-2); 31.1(C-3',4'); 32.6(C-2");
37. 4 (C-5) ; 41. 5 (C.'-4 ) ; 42. 5 (C-l' ) ; 43.2 (C-5' ) ; 45. 1 (C-6) ;
62.7(C-5"); 71.3-72.0(C-3); 97.0-97.9(C-1");
120.5-122.0(C-2); 139.9(C-1).
(5) Synthesis of 1-;5-chloropentyl)-5,5-dimethyl-3-
tetrahydropyranyloxycyclohexene:
1.-(5-Chloropentyl)-5,5-dimethyl-3-tetrahydro-
pyranyloxycyclohexene was obtained in a similar manner to
the step ; 4 ) .
Mass: 317 (M+)
Example 1::
(1) Synthesis of 5,5-dimethyl-3-(14-hydroxytetradecyl)-2-
cyclohexen-l-ol (Compound 1):
17
CA 02348319 2001-05-24
To a solution of magnesium (80 mg, 3.3 mmol) and
iodine (40 mg, 0.1E mmol) in 2 ml of tetrahydrofuran were
added 2 drops of pure 1-(6-chlorohexyl)-5,5-dimethyl-3-
tetrahydropyranyloxycyclohexene. After the reaction was
started, a solution of 1-(6-chl.orohexyl)-5,5-dimethyl-3-
tetrahydropyranyloxycyclohexene (0.98 g, 3 mmol) in 5 ml
of tetrahy'drofuran was added dropwise under reflux
conditions. After 2 hou.rs, to the reaction mixture was
added a liquid containing 1-bromo-8-tetrahydropyranyloxy-
octane (872 mg, 3 mmc.-)l) and dilithium copper tetrachloride
(0.1 M in tetrahydrofuran, 0.3 ml, 0.03 mmol) at 0 C.
After 3 hours, a saturated solution of ammonium chloride
was added at 0 C. The aqueous layer was extracted 3 times
with diethyl ether. The combined organic layers were
washed wit.h a saturated saline, dried over magnesium
sulfate ar.cd then concentrated. The residue was purified
by flash chromatography. The separation of the
tetrahydropyranyl group from the reaction product was then
performed at room temperature for 2 hours in 5 ml of
methanol c:ontaining a catalytic amount (300 mg) of p-
toluenesul.fonic acid. A saturated solution of sodium
carbonate was added to the deprotected product. The
aqueous layer was extracted 3 times with diethyl ether.
The combiried organic layers were washed with saturated
saline, dried over magnesium sulfate and then concentrated.
The residue was purified by flash chromatography to give
5,5-dimethyl-3-(14--hydroxytetradecyl)-2-cyclohexen-l-ol.
18
CA 02348319 2001-05-24
TLC: (dichloromethane/ethyl acetate; 6/4) Rf = 0.37.
1H-NMR (200 MHz; CDC13) ; 6 (ppm) :
0. 88 (s, 3H, -CH3) ; 0. 99 (s, 3H, -CH3) ;
1.25(s,20H,H-11 to H-20); 1.32-1.63(m,4H,H-8,21);
1.64-1.85(m,2H,H-4); 1.86-2.00(m,4H,H-6,7);
3.60-3.67(m,2H,H-22); 3.75-3.90(m,1H,H-1);
5. 42-5. 50 (m, 1H, H-2).
MS (EI) :
337 (M-H, 5) ; 140 (25) ; 139 (C9H15O, 100) ; 109 (C-7H90, 5) ;
55(8).
Compounds 2 to 9 were synthesized in a similar
manner to the step (1).
Incidentally, the data of TLC and IR spectra on the
following Compounds 2 to 11 were the same as those of
Compound 1, and the NMR chemical shifts were substantially
the same as for Compound 1, and so these compounds could
not be distinguished from Compound 1 by these data.
Accordingly, the respective structures were identified by
mass spectroscopy.
(2) 5,5-D.imethyl-3-(18-hydroxytetradecyl)-2-cyclohexen-l-
ol (Compound 2)
Mass: 394 (M+) .
(3) 5,5-Dimethyl-3-(17-hydroxyheptadecyl)-2-cyclohexen-l-
ol (Compound 3)
Mass: 380 (M+).
(4) 5,5-Dimethyl-3-(16-hydroxyhexadecyl)-2-cyclohexen-l-
ol (Compound 4)
19
CA 02348319 2001-05-24
Mass: 366 (M+) .
(5) 5,5-Dimethyl-3-(15-hydroxypentadecyl)-2-cyclohexen-l-
ol (Compound 5)
Mass: 352 (M+).
(6) 5,5-Dimethyl-3-(;13-hydroxytridecyl)-2-cyclohexen-l-
ol (Compound 6)
Mass: 324 (M+).
(7) 5,5-Dimethyl-3-(12-hydroxydodecyl)-2-cyclohexen-l-ol
(Compound 7)
Mass: 310 (M+).
(8) 5,5-Dimethyl-3-(11-hydroxyundecyl)-2-cyclohexen-l-ol
(Compound 8)
Mass: 296 (M+).
(9) 5,5-Dimethyl-3-(10-hydroxydecyl)-2-cyclohexen-l-ol
(Compound 9)
Mass: 282 (M+).
Example 2:
Synthesis of 3-(14-hydroxytetradecyl)-4-methyl-2-
cyclohexen-l-ol (Camp(Dund 10) :
3-(:L4-Hydroxytetradecyl)-4-methyl-2-cyclohexan-l-one
(1 mmol) obtained in accordance with the process described
in WO 99/08987 was dissolved in 2.5 ml of methanol
containing 0.4 M ceri_um trichloride heptahydrate. Sodium
borohydride (1 mmol) was then added to the resultant
solution with stirring. After the mixture was stirred at
room temperature for 5 minutes, the reaction was stopped
by adding 5% hydrochloric acid (1 ml) to the reaction
CA 02348319 2001-05-24
mixture to give 3-(14-hydroxytetradecyl)-4-methyl-2-
cyclohexen-l- ol.
Mass: 324 (M+).
Example 3:
Syntheses of 3--(15-hydroxypentadecyl)-2,4,4-
trimethyl-2-cyclohexen-l-ol (Compound 11):
(1) 2-Acetoxypentadecyl-3,3-dimethyl-l-cyclohexene
obtained in accordance with the process described in WO
99/08987 was oxidized by selenium oxide in a dioxane
solvent urider reflux conditions, or by 70% peroxy-tert-
butanol arid selenium oxide in a water bath to give 3-(15-
hydroxyperitadecyl)--.2,4,4-trimethyl-2-cyclohexen-l-ol.
Mass: 366 (M+) .
(2) 3-(15-Hydroxypentadecyl)-4-methyl-2-cyclohexan-l-
one (1 mmol) obtained.:in accordance with the process
described in WO 99/08987 was dissolved i.n 2.5 ml of a
solution of 0. 4 M CeC:l,j ' 7H20 in methanol. Sodium
borohydride (1 mmo:l.) was then added to the resultant
solution with stirring. After the mixture was stirred at
room temperature for 5 minutes, the reaction was stopped
by adding 5% hydrochloric acid (1 ml) to the reaction
mixture to give 3-(]5-hydroxypentadecyl)-2,4,4-trimethyl-
2- cyclohexen-l-ol.
Test Example 1: Neurocyte-growing effect
Fetal rat cerebral hemisphere derived neurons (15
days old) were used, and culture was carried out in
accordance with the method by Borg et al. [Borg J., et al.,
21
CA 02348319 2001-05-24
Dev. Brair., Res., 18, 37 (1995)]. Cells dissociated were
plated at a density of' 1.5 x 105 cells/dish in polylysine-
coated dishes 35 mm._in diameter. A DMEM medium
(containirig insulin, transferrin, progesterone, sodium
selenate and putesc:ine; 3 ml) and Compourid 1(10-9 to 10-5
M in methanol) were added to the respective dishes. The
cells were cultured for. 2 days without changing the medium.
After the culture, the number of cells was counted through
a microscope. Incidentally, 0.1 ethanol was used as a
control. The results are diagrammatically illustrated in
Fig. 1(means of 3 independent experiments).
The compound according to the present invention
significaritly exhibited a neurocyte-growing effect at 10-6
M.
Test Example 2: Neurite-outgrowing effect
Cells were ctiltured in the same manner as in Example
1. The cultured celis were immobilized by 4%
paraforma__dehyde, and neurons were observed through a
microscope by immunostaining making use of an antibody to
measure neurite length by image analysis. The results are
diagrammatically illustrated in Fig. 2.
The compound according to the present invention
exhibited an excellent neurite-outgrowing effect in a low
dose.
22
