Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
~ 37 RAN 4212/31
5The present invention is concerned with novel chole-
calciferol derivatives of formula
10H3C", ~\~-C H3
~ ~ C H3
15 ,~ H
1~
CH2
HO' ~ OH
The invention is also concerned with pharmaceutical
25 preparations on the basis of the eompounds of formula I, a
process for the manufac-ture of these compounds and lnter-
mediates occuring in said process.
The eompounds of formula I are manufactured in
30 aeeordance with the invention by thermally isomerising a
preeholecalciferol derivative of formula
Mé/30.7.82
3'~
, F
=C H3
l l A
o
In the formulae presented herein a wavy line between
- a C atom and a substituent indicates that the absolute
',: configuration at the C atom is R or S, or that the corres-
16 ponding compound is in form of a R,S-mixture.
The thexmal isomerization of a compound of formula II-A
can be carried out by heating in an inert sol~ent, such as
an ether, e.g. dioxane, tetrahydrofuran or dimethoxyethane;
~, 20 an aromatic hydrocarbon, eOg. benzene or toluene, under an
inert atmosphere~such as argon or helium, by methods well
known in the art, see e.g. J.A.C.S. 98 (1973) 2748.
\
As used hereinafter, the term "lower" refers to groups
~ 26 containing l to 8 carbon atoms. Examples of lower alkyl
y: groups are methyl, ethyl, n-propyl, i-propyl, tert.-butyl,
i hexyl, heptyl and octyl. Examples of lower alkylene groups
are methylene, ethylene and propylene. Examples of lower
alkoxy groups are methoxy, ethoxy, isopropoxy and tert.-
30 butoxy. Examples of lower alkanoyloxy groups are formyloxy,
acetoxy, butyryloxy and hexanoyloxy. The term "substituted"
as applied to ;'phenyl" refers to phenyl which is substituted
with one or more groups; such as lower alkyl, fluoro,
chloro, bromo, iodo, nitro, cyano or trifluoromethylO
/~ The compounds of formula II-A can be manufactured by
i; reacting a compound of formula
. ,
, . .
"
-- 3
X~CH3
OY
CH3
wherein X is iodo, bromo, chloro, lower alkylsulfonyl-
oxy, phenylsulfonyloxy or subs-tituted phenylsulfonyl-
oxy, and Y is a group of the formula
Rl
R - O - C -
R2
wherein R is hydrogen or lower alkyl, R and R are
lower alkyl or R and R taken together are C3 6-
j5 alkylene,
with a metallated pregn-5-en-21-oic acid ester of formula
M ~ C02~4
YO ~
. CH H
~ ~ IV A
25 yoJ`~J
wherein M is lithium, sodium, potassium, magnesium~2
or zinc/2, R is lower alkyl, phenyl or substituted
phenyl and Y is as above,
according to the method described in J.Chem.Soc., Perkin
Trans. I, 1282 ~1978),to give the corresponding alkyla-ted
compound of formula
35~
-- 4
YO
wherein R and Y are as above.
This reaction may be carried out in an aprotic inert
solvent, such as an ether, e.g. tetrahydrofuran; or an
15 amlde, e.g. hexamethylphosphoramide, conveniently at a
temperature between -78 and 60C. The product of formula V-A,
containing the desired 20R-absolute confiyuration, can be
isolated by the usual chemical and physical means.
The compound of formula V-A is then reduced to the
alcohol of formula
F
HOH2C" ~ CH3
CH3 1 .H~ \
Y~ 1/\/~ CH3 VB
J I!~
y
.
wherein Y is as above,
35 by reactio~ with a complex metal hydride reducing agent,
such as an alkali metal aluminum hydride, e.g. lithium
aluminum hydride, a mono-, di- 'or tri-(lower alkoxy) alkali
metal aluminum hydrides, e.g. lithium tris(tert.-butoxy)-
,
537~
-- 5
aluminum hydride; a mono-, di- or tri(lower alkoxy-lower
alkoxy) alkali metal aluminum hydrides, e.g. sodium bis(2-
methoxyethoxy)aluminum hydride; or a di(lower alkyl)
aluminum hydrides, e.g. diisobutyl aluminum hydride. ~ui-
table solvents for this reduction include ethers, such asdiethyl ether. The reduction is conveniently carried out at
a temperature between about 0C and 100C.
The alcohol of formula V-B is then converted in a
10 manner known per se to the halide or sulfonate of formula
X H2C" ~"~C H3
YO ~--~ C H3 V C
YO~
wherein ~ and Y are as above.
To prepare a sulfonate of formula V-C, the compound of
formula V-B is reacted with a sulfonyl halide in the
presence o~ a base. The preparation of a halide of formula
V-C can be accomplished either by reacting the alcohol of
formula V-B with a halogenating agent, such as phosphorus
30 tribromide, or by reaction of a sulEonate ester of formula
V-C e.g~ a tosylate, with a halide ion containing compound~
such as an alkali metal bromide or iodide, e.g. potassium
bromide.
The compound of formula V-C is then conver-ted to the
compound of formula
-- 6
H3C~" ,/\ ~ C H3
r~3~ CH03Y
YO~
wherein Y is as above,
with a complex metal hydride reducing agentO
The reduction is normally carried out with one of the
15 reducing agents and i.n one of the solvents mentioned above
in connection with the reduction of a compound V-A to a
compound V-B, at a temperature between about room temperature
and 100C. Other suitable reducing agents, particularly
when ~ is iodo or bromo, are alkali metal cyanoborohydrides
20 such as, sodium cyanoborohydride (sodium cyanotrihydro-
borate); tri (lower alkyl) tin hydrides, such as tri-n-
butyltin hydride; and tri (aryl) tin hydrides, such as
triphenyltin hydride. The reduction is normally carried out
at a temperature between about -20 and 80C.
The compound of ~ormula V-D is then conver-ted -to the
lc~,25-dihydroxy-24-fluorocholesterol of formula
$
HO ~ H3 VE
l H H
HO ~ ~
-- 7
by removal of the protecting groups Y wi-th a strong acid in
a protic solvent. Suitable strong acids include mineral
acids, such as hydrochloric acid, and organic sulfonic acids,
such as p-toluenesulfonic acid. Suitable solven-ts include
alcohols, such as methanol, and water containing a miscible
co-solvent -to help solubilize the organic reactants, such
as ethers, e.g. -tetrahydrofuran; or a ketone, such as ace-
tone. The reaction is preferably carried out at a tempe-
rature be-tween about -lO and 80C.
The cholesterol of formula V-E is -then alkanoylated -to
the la,3~-dialkanoate of formula
RS0 ~ CH3
~--
R5 O~J
wherein R is lower alkanoyl,
25 by methods well known in the art.
The compound of formula VI-A is allylically halogena-ted
to a mixture of 7a- and 7~-halocholesterols of formula
F
H3C", ~,_C H3
R o ~ \C~H03~
~ VI B
R50~ ~`
5~
-- 8 --
wherein R is as above and Z is iodo, bromo or chloro,
using a halogenation agent, such as N chloro- or N-bromo-
succinimide, dissolved in a sa-tura-ted aliphatic hydrocarbon
or halocarbon, such as hexane, in the presence of an acid
scavenger, such as sodium bicarbonate, at the boiliny poin-t
of the reaction medium.
Without separation of the 7~-halo-isomer from the pre-
dominant 7c~-isomer, the compound of formula VI-B is dehydro-
10 halogenated to the steroid-5,7-diene of formula
H3C", ~ CH3
~5 ~ ~ CH3
R5
wherein R is as above,
utilizing a heteroaromatic tertiary amine, such as a pyri-
dine, e.g. picolines, lutidines and collidines; or an ali-
25 phatic tertiary amine, e.g. triethylamine, tripropylamine,1,5-diazabicyclo(4.3.0) non-5-ene or 1,4-diazabicyclo(2.2.2)
octane. Trialkylphosphi-tes are also useful in the dehydro-
halogenation step. Suitable inert organic solvents include
aromatic and aliphatic solvents, such as xylene. The reac-
30 tion proceeds readily at temperatures from abou-t 50C to
the reflux temperature of the reaction medium. The steroid
5,7-diene oE formula VI-C can be isolated by the usual chemi-
cal ancl physical means, such as chromatography, and in this
manner can be separated from any undesired impurities, such
35 as the correspondiny 4,6-diene.
- 9
The steroid 5,7-diene of formula VI-C is converted to
the precholecalciferol-l~,3~-dialkanoate of formula
` H3C", ~ CH3
R50 ~ ~ H3
CH3 ~\ ~ 13 B
R50 ~ H
wherein R5 is as above,
16 by irradiation under an inert atmosphere, such as nitrogen,
at a temperature of about -40 to +25~C. Suitable sources
of irradiation include high and low pressure mercury lamps,
Suitable inert organic solvents for the irradiation include
mixtures of saturated aliphatic hydrocarbons and ethereal
20 solvents, such as mixtures of hexane and tetrahydrofuran.
~ pon completion of the irradiation, the solvants are
removed by evaporation and the residue i~ separated into
the pure precholecalciferol la,3~-dialkanoate of formula
~5 II-B and pure unchanged steroid 5,7-diene of formula VI-C
by liquid chromatography using a solid absorbent, e.g.
Porasi~, and an inert organic eluant. Suitable eluants
include mixtures of hydrocarbons and esters, such as a
mixture of n-hexane and ethyl acetate.
Unchanged 5,7-diene of formula VI-C is recycled
through the irradiation process to obtain additional
quantities of pure precholecalciferol l~,3~-dialkanoate of
formula II-B.
.
The compound of formula II-B is then saponified to
the compound of formula II-A with a strong base in a protic solvent.
Suitable bases include alkali and alkaline earth hydroxides
`- *Trade mark
.
-- 10 --
or alkoxides, such as potassium hydroxide. Suitable sol-
vents include alcohols, such as methanol, and water con-
taining a miscible co-solvent to help solubilize the
organic reactan-ts, such as an ether, eOg. tetrahydrofuran.
It is preferable to carry out the removal of the alkanoyl
groups R at a temperature between about -20 and 60C and
under an inert atmosphere, e.g. nitrogen.
The compounds of formula III can be prepared as shown
10 in the scheme below, wherein R and Y have the meaning given
above, and as described in -the Examples.
~0 ~ H~ CH3
IIIA IIIB illC
25 R50/\/~011 C~3 HO ^~ '~CH
5~
The metallated compounds of formula IV-A can be pre-
pared by treating a compound of formula
~Co2R4
IV B
YO
wherein R4 and Y are as above,
1S with an appropriate organome-tallic reagent, for example
lithium diisopropylamide, sodium hexamethyldisilazane or
potassium hydride.
The compounds of formula IV-B can be prepared as shown
20 in the sheme below, wherein Y and R4 are as above, and as
described in the Examples
H
IVE I Co2R4
30yo ~ 4
YOl,~J lV-B
The compounds of formulae II-A and II-B, i.e. of
- formula
3~
- 12 - F
H3C~",
R o~'--~ C 03H
R6
wherein R is hydrogen or lower alkanoyl,
10 those of formulae VI-A, VI-B and VI-C, i.e. of formula
H3C-" ~--~C H3
CH3 OH
15 R O f~/~ C H3
z,
wherein Q is hydrogen and Z'is hydrogen, iodo, bromo
or chloro, or Q and Z'taken together are an additional
7,8-bond and R is above,
25 those of formulae V-A, V-B, V-C, V~D and V-E, i.e. of
formula
30 X 1,. ~ C H3
~ ~ C H3
CH H
~`J
¦ I H I H
YoJ~ ~J
wherein Y' is hydrogen or a group Y and X' is methyl,
or Y' is a group Y and X' is -CH2X, -CH2OH or -COOR ,
and Y, X and R are as above,
those of formula IV-A and IV-B, i.e. of formula
M ~ Co2R4
YO
YO~
wherein M' i5 hydrogen or a group M, and M, R and Y
are as above,
as well as those of formulae 2, 3, III-A, III-B, III-C and
III are novel and also form part of the present invention.
The compounds of formula I are characterized by -the
ability to increase intestinal calcium transport, to
increase serum calcium and phosphate concentrations and to
increase the deposition of these minerals in bones. These
compounds will find application as substitutes for la,25-
25 dihydroxycholecalciferol in the treatmen-t of disease states
characterized by metabolic calcium and phosphate deficien-
cies, such as osteomalacia, osteoporosis, rickets, osteitis
fibrosa cystica, renal osteodystrophy, osteosclerosis,
convulsions, osteopenia, fibrogenesis imperfecta ossium,
30 secondary hyperparathyroidism, hypoparathyroidism, hyper-
parathyroidism, cirrhosis, obstructive jaundice, dru~
induced metaholism, medullary carcinoma, chronic renal
disease, hypophosphatemic vitamin V-resistant rickets,
vitamin V-dependen-t rickets, sarcoidosis, glucocorticoid
35 antagonism, malabsorp-tion syndrome, stea-torrhea, tropical
sprue, idiopatic hypercalcemia and milk fever.
3t7
- 14 -
la,25-Dihydroxy--24R-fluorocholecalciferol has shown
activity in the anti-rachitoyenic test in chicks and has
also shown a duration of activity in stimulation of
intestlnal calcium absorption of 24 hours after a single
oral 100 nanogram dose, comparable to that of la,25-
dihydroxycholecalciferol.
White Leghorn chicks are placed on a vitamin D-clefi-
cient diet which contains 1% calcium and 0.7% phosphorous
and are housed under ultraviole-t-free lighting. The test
compounds are dlssolved in propy]ene glycol and admini-
stered orally for 21 consecutive days to chicks which are
one to two days of age at the start of treatment. Con-trols
are treated with vehicle alone. Chicks are autopsied on the
15 day after the last treatment day and -tibia ash weigh-twas
determined. Nine or ten chicks are used for each treatment
group and for the control group. The results of -the la,25-
dihydroxy-24R-fluorocholecalciferol (compound X) anti-
rachitogenic activity assay are shown in Table I along
20 with comparative data showing the anti-rachitogenic acti-
vity of 24,24-difluoro-la,25-dihydroxycholecalciferol
(compound A). The results show that la,25-dihydroxy-24R-
fluorocholecalciferol possesses potent anti-rachitogenic
activity.
In an other experiment the chicks are placed on a
vitamin D-deficient diet and housed under ultraviolet-free
lighting for 21 days. In order to determine the effects
of compound X on intestinal calcium absorption, a single
30 oral dose dissolved in propylene glycol is administered.
At various times after dosing, 2 ~Ci of Ca (chloride~
is given orally and serum radioactivity is measured 45 minu-
tes after administration of -the isotope. Vehicle-trea-ted
con-trols are included at each time period. Ten chicks are
35 used in each treatment and control group. The results
given in Table II, show that la,25-dihydroxy-24R-fluoro-
cholecalcifero] has intestinal calcium absorption activity
of about 2~ hours duration.
37
- 15 -
Table I
Dose ~ean tibia ash weight (mg)
ng/chick/day Compound
A X
120.9+8.3
3 192.2-~ 6.2 293.3+ 4.R
258.6+10.0 330.4+ 7.7
229.8+14.2 337.9+10.6
Table II
Treatment Time Serum 45Ca
(hours) _ epm/0.2__ml
15 0.2 ml vehiele 24 408+30
eompound X 793+73 *
0.2 ml vehiele 48 439+30
eompound X 520+38 NS
0.2 ml vehiele 72 398-~31
20 eompound ~ 372-~39 NS
* - statis-tieally signifieant result
NS = not statistieally signifieant
la,25-Dihydroxy-24R-fluoroeholeealeiferol ean be
administered in dosages that are in the range of about
0.10-3.0, preferably 0.25-2.0 mierograms per day for the
treatment of such disease states as osteodystrophy, steroid
induced osteopenia, hypoparathyroidism, hypophosphatemic
30 rickets and hypophosphatemic osteomalacia, whieh are
eharaeteri~ed by lower than normal levels of endogenously
produeed la,25-dihydroxycholecalciferol. la,25-Dihydroxy-
24R-fluorocholeealeiferol ean be administered orally,
subeutaneously, intramuscularly, intravenouslv, intra-
35 peritoneally or topically and ean be formulated into com-
positions, sueh as tablets, capsules or elixirs for oral
administration, or in sterile solutions or suspensions ~or
parenteral administration for the treatment of the afore-
- 16 -
mentioned disease states. The amount of active substance
in the foregoing composi-tions is in the range previously
indicated. Illustrative of the adjuvants which may be
incorporated Lnto the above compositions are the following:
a binder such as gum tragacanth, acacia, corn starch or
gelatin; an excipient such as calcium phosphate; a disin-
tegra-ting agent such as corn or potato starch or alginic
acid; a lubricant such as magnesium stearate:, a sweetening
agen-t such as sucrose, lactose or saccharin; a flavoring
agent such as peppermint, oil of wintergreen or cherry.
Various other materials may be present as coa-ting or to
o-therwise modify the physical form of the dosage unit. For
instance, tablets may be coated with shellac, sugar or
both. A syrup or elixir may contain the active compound,
sucrose as a sweetening agent, methyl and propyl parabens
as preservatives, a dye and a flavoring such as cherry
or orange flavor.
la,25-Dihydroxy-24R-fluorocholecalciferol can be
administered for the treatment of milk fever in pregnant
ruminant animals prior to parturation in dosages in the
range of 100-1500 micrograms/day using conventional
formulations.
Sterile compositions for injection an~/or topical
administration can be formulated according -to conventional
practice by dissolving or suspending 1~,25-dihydroxy--24R-
fluorocholecalciferol in a vehicle such as a 10-20%
ethanol-water mixture, a 10-20% propylene glycol-water
30 mixture, a naturally-occurring vegetable oil, such as
sesame oil, peanut oil, cottonseed oil or a synthetic
fatty vehicle, such as ethyl oleate. For example, a suitable
formulation for intravenous injection would be 2-3 ml of
a 10-20% ethanol-water solution or a 10-20% propylene
35 glycol-water mixture containing 200-1000 micrograms of
1~,25-dihydroxy-24R-fluorocholecalciferol. Exemplary of a
sui-table formulation for topical administration would be a
vegetable oil solution or suspension containing 200-1000
- 17 ~
mlcrograms of 1~,25-dihydroxy-24R-fluorocholecalciferol.
la~2s-Dihydroxy-24R-fluorocholecalciferol can also
be formulated for oral adminis-tration hy incorpora-tion of
100-1500 micrograms thereof into fatty acid pellets.
la,25-Dihydroxy-24R-fluorocholecalciferol may also
be formulated for intramuscular injection by suspension
of 100-1500 micrograms of la,25-dihydroxy-24R-fluoro-
cholecalciferol in a vehicle such as a vege-table oil, an
ethanol-water solution or a propylene glycol-water solu-
tion, containing from 80-95% ethanol or propylene glycol.
;D5~7
- 18 -
_xample 1
A. A mixture of 0.104 g (0.00024 mole) of [1~,3~,6Z,24R]-
24-fluoro-9,10-secocholesta~5(10),6,8--trien-1,3,25-triol
and 18 ml of p-dioxane was heated a-t reflux (100C) for
1 hr and then cooled. The mixture was evaporated to
dryness. The residue was purified hy column chromatography
on silica gel with 3:1 ethyl acetate-hexane as eluant~-to
give [la,3~,5Z,7E,24R]-24-fluoro-9,10-secocholesta-5,7,-
-lO 10(19)-trien-1,3,25-triol (1~,25-dihydroxy-24R-fluoro-
cholecalciferol), [a]D +68 (c = 0.5, r~eOH).
B. The star-ting secocholestane can be prepared as follows:
a) To a solution of 1.2 ml of diisopropylamine in 4 ml of
tetrahydroEuran at -30C was added 4~5 ml (0.0072 mole) of
1.6M of butyllithium in hexane. After stirring for 0.5 hr,
3.57 g (0.0065 mole) of [la,3~]-1,3-bis[(-tetrahydro-2H-
pyran-2-yl)oxy]-pregn-5-en-21-oic acid e-thyl ester in 36 ml
of tetrahydrofuran was added dropwise. The mixture was
stirred for 1 hr at -30C and cooled to -70C. A solution
of 3.40 g (0.011 mole) of 4-(1-ethoxyethoxy)-3R-fluoro-1-
iodo-4-methy]pentane in 6 ml of hexamethylphosphoramide was
added dropwise. The mixture was stirred at -70C for 1 hr
and allowed t:o warm to 25C and stir for 1 hr. The mixture
was diluted with 9:1 hexane-ether. The solution was washed
with water and saturated brine. The organic phase was dried,
filtered and evaporated to dryness. The residue was puri-
fied by column chromatography on silica gel to give
[la,3~,24R]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-25-(1-
ethoxyethoxy)--24-fluoro-cholest-5-en-21-oic acid ethyl
ester, [a]~ +18 (c = 1, CHC13).
b) To a mixture of 0.34 g (0.0090 mole) of lithium alu-
35 minum hydride and 17 ml of tetrahydrofuran at 0C was added
4.26 g of [la,3~,24R]-1,3-bis[(tetrahydro-2H-pyran-2-yl)-
oxy]-25-(1-ethoxyethoxy)-24-fluorocholest-5-en-2l-oic acid
-- 19 --
ethyl ester in 60 ml of tetrahydrofuran. The mixture was
heated at 50~C for 1.5 hr, cooled to 0C and diluted with
200 ml of ether. The mixture was quenched with 0.68 ml of
water ancl 0.55 ml of 10% aqueous sodium hydroxide. The
mixture was stirred at 25C for 0.5 hr and filtered. The
solids were triturated with ether and filtered. Evaporation
of solvent afforded [la,3~,24R]-1,3-bis[(tetrahydro-2H-
pyran-2-yl)oxy]-25-(1-ethoxyethoxy)-24-fluorocholes-t-5-en-
21-ol, [a]D +7 (c = 1.1, CHCl3).
c) A mixture of 3.99 g (0.0058 mole) of [la,3~,24R]-1,3-
bis[(tetrahydro-2H~pyran-2-yl)oxy]-25~ ethoxyethoxy)-24-
fluorocholest-5-en-21-ol, 15 ml of pyridine and 2.20 q
(0.0115 Mole) of p-toluenesulfonyl chloride was stirred at
15 0C for 18 hr. The mixture was quenched with ice chips.
The mixture was then poured into water and extracted with
methylene chloride. The organic phase was washed with 10%
aqueous sulfuric acid and saturated sodium bicar-
bonate solution. The organic layer was dried, filtered and
20 evaporated to dryness to yield [la,3~,24R]-1,3-bis[(te-tra-
hydro-2H-pyran-2-yl)oxy]-25-(1-ethoxyethoxy)-24-fluoro-
cholest-5-en-21-ol 21-(4-methylbenzenesulfonate), [a]20 ~8
(c = 1, CHCl3).
25 d) A mixture of 0.223 g (0.00026 mole) of [la,3~,24R]-
1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-25-(1-e-thoxye-thoxy)-
24-fluorocholest-5-en-21--ol 21-(4-me-thylbenzenesulfonate)
and 0.150 g (0.0010 mole) of sodium iodide in 2 ml of ace-
tone was heated at 50 for 18 hr and cooled. The mixture
30 was poured into water and the product was isolated with
methylene chloride. The organic layers were washed with
aqueous sodium sulfite solution and saturated sodium
bicarbona-te solution. The organic layers were dried, filte-
red and evaporated to dryness to yield [la,3~,24R]-1,3-
35 bis[(tetrahydro-2H-pyran-2-yl)oxy]-25-(1-ethoxyethoxy)-24-
fluoro-2l-iodocholest-5-ene .
a~
-20
e)1) A mixture of 0.710 g (0.0187 mole) of lithium aluminum
hydride, 80 ml of tetrahydrofuran and 4.85 g (0.0057 mole)
of [la,3~,24R~-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-25-
(1-ethoxyethoxy)-24-fluorocholest-5 en-21-ol 21-(4-me-thyl-
benzenesulfonate) was heated at reflux (60C) for 1 hr andcooled to 0C. The mix-ture was diluted with 200 ml of
ether and quenched with 1.4 ml of water and 1.1 ml of 10~
aqueous sodium hydroxide solution. The mixture was stirred
for 1 hr and filtered. The solids were triturated with
10 ether and filtered. The combined filtrates were evaporated
to dryness and chromato~raphed on silica gel to yield
[la,3~,24R]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-25-(1-
ethoxyethoxy)-24-fluorocholest-5-ene, Ca]D +7 (c = 1,
CHCl3).
e)2) By an al-ternative procedure, a mixture of 0.206 g
(0.00025 mole) of [la,3~,24R]-1,3-bis[(tetrahydro-2H-pyran-
2-yl)oxy]-25-(1-ethoxyethoxy)-24-fluoro-21-iodocholest-5-ene,
0.087 g (0.00030 mole) of tri-n-butyltin hydride and 3 ml
20 of tetrahydrofuran was stirred at 25C for 18 hr under an
argon atmosphere. The mixture was evaporated -to dryness
and the residue was purified by chromatography on silica
gel to yield [la,3~,24R]-1,3-bis-[(tetrahydro-2H-pyran-2-
yl)oxy]-25-(1-ethoxye-thoxy)-24-fluorocholest-S-ene,
25 [a]D ~7 (c = 1, CHCl3).
f) A mixture of 3.56 g (0.0053 mole) of [la,3B,24R]-1,3-
bis-[(tetrahydro-2H-pyran-2-yl)oxy]-25-(1-ethoxyethoxy)-
24-fluorocholest-5-ene, 140 ml of methanol and 0.58 g of
30 p-toluenesulfonic acid monohydrate was stirred a-t 25C for
3 hr. The mixture was quenched by adding 1.0 g of sodium
bicarbonate and stirring for 0.5 hr. The mixture was eva-
porated to dryness. The residue was triturated with ethyl
acetate, filtered and evaporated to dryness~ The crude
35 solid was recrystallized from ethyl acetate -to yield
[la,3~,24R]-24-fluorocholest-5-en-1,3,25-triol, mp 156-158C,
[a]D +4 (c = 0.5, MeOH).
- 21 _
g) A mixture of 1.53 g (0.0035 mole) of [la,3~,24R]-24-
fluorocholest-5-en-1,3,25-triol, 13 ml of pyxidine and 6 ml
of acetic anhydride was stirred at 0C for 1 hr and at
25C for 17 hr. The mixture was diluted with 20 ml of
methanol at 0C and evaporated to dryness. The residue was
dis5solved in methylene chloride. This solution was washed
with 10% aqueous sulfuric acid and saturated sodium bicar-
bonate solution. The organic phase was dried, filtered
^ and evaporated to dryness to yield [la,3~,24R]-24-fluoro-
10 cholest-5-en-1,3,25-triol 1,3-diacetate, [a]23 _7~ (c = 1,
- CHC13).
. .
~? h) A mixture of 0.520 g (0.0010 mole) of [la~3B~24R]-24
fluorocholest-5-en-1,3,25--triol 1,3-diacetate, 0.45 g of
15 sodium bicarbonate, 0.192 g (0.00066 mole) of 1,3-dibromo-
~;' 5,5-dimethylhydantoin and 25 ml of hexane was heated at
;~i reflux ~80C) for 1 hr and cooled. The mixture was filtered
and the solids were triturated with hexane and filtered.
;ê The filtrates were evaporated to dryness to yield [1,3~,-
~` 20 7~,24R]-7-bromo-24-fluorocholest-5-en-1,3,25-triol 1,3-
diacetate.
~i
i) A mixture of 0.605 g (0.001 mole) of [1~,3~,7~,24R]-
.~ 7-bromo-24-fluorocholest-S-en-1,3,25-triol 1,3-diacetate,
25 0.6 ml of s~collidine and 18 ml of xylene was heated at
reflux (140C) for 0.5 hr and cooled. The mixture was
diluted with 30 ml of toluene. This solution was washed
~`~ with 10% aqueous sulfuric acid and saturated sodium
bicarbonate solution. The organic phase was dried, filtered
30 and evaporated to dryness. The residue was purified by
~; col~mn chromatography on silica gel to yield [la,3~,24R~-
24-fluorocholesta-5,7-dien-1,3,25-triol 1,3 diacetate,
[aJD -215 (c = 0.5, CHCl3).
36 j) A mixture of 0.258 g (0.0005 mole) of [1~,3~,24R]-24-
fluorocholesta-5,7-dien-1,3,25-triol 1,3-diacetate in 40 ml
of hexane and 10 ml of tetrahydrofuran was irradiated for
0.5 hr under argon at -5C using a 450W Hanovia high
*Trada mark
- 22
pressure mercury lamp, cooled with a Vycor-glass cooling
finger. The solution was evaporated to dryness and the
residue was purified~y ~1~ chromatography on silica gel
with a 4:1 mixture of n-hexane-ethyl acetate as eluant~to
give [la~3~6z~24R]-24-fluoro-9~lo-secocholesta-5(lo)~6~8
trièn-1,3,25-triol 1,3-diacetate.
k) A solution of 0.125 g (0.00024 mole) of [la,3~,6Z,24~]-
10 24-fluoro-9,10-secocholesta-5(10),6,8-trien_1,3,25_triol
1,3-diacetate and 15 ml of 1% potassium hydroxide in
methanol was stirred at 0C for 4 hr. The mixture was neu-
tralized to pH 7.5 with glacial acetic acid in methanol.
The solution was evaporated to dryness at 0C. The residue
15 was partitioned between ethyl acetate and water. The organic
phase was washed with saturated brine and dried. The mix-
ture was filtered and evaporated to dryness to yield
[la,3~,6Z,24R]-24-fluoro-9,10-secocholesta-5(10),6,8-trien-
1,3,25-triol.
~0
C. The intermediate, 4-(1-ethoxyethoxy)-3R-fluoro-l-iodo-
4-methylpentane, can be prepared as follows:
,
a) To a solution of 16.5 g (0.102 mole) of diethylamino-
25 sulfur trifluoride in 20 ml of methylene chloride
at -70C was added dropwise 3.50 g (0.034 mole) of (-)-2S-
, hydroxy-4-butyrolactone in 30 ml of methylene chloride.
The mixture was stirred at -70C for 1 hr and was warmed
- to 25C and stirred 1 hr. The mixture was poured into
30 saturated sodium bicarbonate solution at 0C. The
product was extracted with methylene chloride. The organic
phase was dried, filtered and evaporated to dryness. The
residue was purified by column chromatography on silica
gël to yield 2R-fluoro-4-butyrolactone, [a]D +50 (c = 1,
s 3S C~C13).
.. . .
*Trade Mark
,
..
- 23 _
b) To a solution of 15.8 ml of 1~5M e-thereal me-thyl-
lithium (0.025 mole) at 0C was added dropwise 1.05 g
(0.010 mole) of 2R-fluoro~4-butyrolactone in 35 ml of ether.
The mix-ture was s-tirred a-t 0C for 1 hr and a-t 25C for
1 hr. The mixture was quenched by adding 2 ml of saturated
brine at 0C. The mix-ture was poured in-to saturated brine
and the product was isolated with ether. The e-~her layers
were dried, filtered and evaporated -to dryness to give
4-methyl-3R--Eluoro-1,4-pentanediol, [~]2Z +39 (c = 1,
10 CHCl3).
c) To a mixture of 0.540 g (0.0040 mole) of 4-methyl-3R-
fluoro-1,4-pentanediol and 3 ml of pyridine at 0C was
added 2 ml of acetic anhydride and the mixture was stirred
15 at O~C for 1 hr and at 25C for 1 hr. The mixture was
added to 10 ml of me-thanol and the solution was evaporated
to dryness to yield 4-methyl-3R-fluoro-1,4-pentanediol
1-acetate, ~a]~ +49 (c = 0.5, CHC13).
20 d) A mixture of 2.48 g (0.014 mole) of 4-methyl-3-R-Eluoro-
1,4-pentanediol 1-acetate, 65 ml of ethyl vinyl e-ther and
0.25 g of p-toluenesulfonic acid monohydrate was stirred
at -70C for 1 hr. The mixture was quenched by adding 8 ml
of triethylamlne and evaporated to dryness. The residue
25 was taken up in ether. This solution was successively
washed with satura-ted sodium bicarbonate solu-tion
and saturatecl brine. The ether phase was dried, fil-tered
and evaporated to dryness to yield an oil containing 4-(1-
ethoxyethoxy)-3R-fluoro-4-methyl-1-pentanol 1-aceta-te.
e) To a mixture of 0.79 g (0.0~1 mole) of lithium aluminum
hydride in 40 ml of ether at 0C was added dropwise 3.48 g
(0.014 mole) of 4-(1-ethoxyethoxy)-3R-fluoro-4-methyl-1-
pentanol 1-acetate in 100 ml of ether. The mixture was
35 heated at reflux for 3 hr and then was cooled to 0C. The
mixture was quenched by adding dropwise 1.5 ml of water
followed by 1.2 ml of 10% aqueous sodium hydroxide. The
mixture was stirred at 25C for 0.5 hr and was filtered.
- 24 -
The solids were triturated with e-ther and Eiltered. Eva-
poration of solvent and column chromatography of the
residue on silica gel afforded 4~ ethoxyethoxy)-3R-
fluoro-4-methyl-1-pentanol, [a]D ~18 (c = 1, CHCl3).
f) A mixture of 3.34 g (0.016 mole) of 4-(1-ethoxyethoxy)-
3R-fluoro-4-methyl-1-pentanol, 12 ml of pyridine and 4.67 g
(0.024 mole) of p-toluenesulfonyl chloride was stirred at
0C for 18 hr. The mlxture was quenched with ice chipsJthen
10 poured into water and extracted with methylene chloride.
The organic phase was sequen-tially washed with 10% aqueous
sulfuric acid and saturated sodium bicarbonate
solution. The organic phase was dried, filtered and eva-
porated to dryness to yield oily 4-(1-ethoxyethoxy)-3R-
15 fluoro-4-methyl-1-pentanol 1-(4-methylbenzenesulfonate).
g) A mixture of 5.42 g (O.OlS mole) of 4-(1-ethoxyethoxy)-
3R-fluoro-4-methyl-1-pentanol 1-(4-methylbenzenesulfonate),
70 ml of acetone, 1 ml of diisopropyle-thylamine and 30.3 g
20 (0.202 mole) of sodium iodide was stirred at 25C for 18 hr.
The mixture was evaporated to dryness. The residue was
partitioned between 5% aqueous sodium sulfite solu-tion and
methylene chloride. The organic phase was washed with
saturated sodium bicarbonate solution, then dried,
25 filtered and evaporated to dryness to yield 4-(1-ethoxy-
ethoxy)-3R-fluoro-1-iodo-4-methylpentane, Ca]D4 +43
(c = 1, CHC13).
D. The intermediate, [1~,3~-1,3-bis[(tetrahydro-2H-pyran-
30 2-yl)oxy]-pregn-5-en-21-oic acid ethyl ester, can be pre-
pared as follows:
a) A mixture of 0.91 g (0.0030 mole) of 1~,3~-dihydroxy-
androst-5-en-17-one, 15 ml of tetrahydrofuran, 1.26 g
35 (0.015 mole) of 3,4-dihydro-2H-pyran and 0.028 mg of
p-toluenesulfonic acid monohydrate was stirred at 25C for
18 hr. The mixture was diluted with methylene chloride.
~ a ~ '7
- 25 _
This solution was then washed with sa-turated s~dlum
bicarbonate solution. The organic phase was dried, filtered
and evaporated to dryness to yield [la,3~]-1,3-bis-~(tetra-
hydro-2H-pyran-2 yl)oxy]andros-t-3-en-17-one, [a]D +34 3
(c = 1, CHCl3).
b) To a mixture of 1.00 g (0.0021 mole) of [la,3~]-bis-
[(tetrahydro-2H-pyran-2-yl)oxy]andros-t-3-en-17-one, 1.94 g
(0.0087 mole) of triethyl phosphonoacetate and 14 ml of
ethanol was added 0.68 g (0.010 mole) of sodium
ethoxide in 7 ml of e-thanol. The mixture was stirred at
reflux for 18 hr and cooled. The mixture was concentrated
under reduced pressure, then partitioned between water and
ether and the organic phase was washed with saturated brine.
15 The organic phase was dried, filtered and evaporated to
dryness. The residue was chromatographed on silica gel to
yield [la,3~,17(20)E]-1,3-bis[(tetrahydro-2H-pyran-2-yl)-
oxy]pregna-5,17(10)-dien-21-oic acid ethyl ester, [a]20 -8
(c = 1, CHC13).
c) A mixture of 0.32 g (0.00059 mole) of [la,3~,17(20E]-
1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]pregna-5,17(20)-dien-
21-oic acid ethyl ester, 0.10 g of pla-tinum oxide and 20 ml
of ethanol was stirred in 1 atmosphere of hydrogen for 2 hr.
25 The mixture was fil-tered through diatomaceous earth and
the solids were washed with e-thanol. The combined filtrates
were evaporat:ed to dryness to yield [la,3~-1,3-bis[(tetra-
hydro-2H-pyran-2-yl)oxy]pregn-5-en-21-oic acid ethyl ester,
[a]D ~]3 (c = 1, CHC~3).
Example 2
A. In a manner similar to that described in Example lA
[la,3~,6Z,24S]-24-Eluoro-9,10-secocholes-ta-5(10),6,8-
trien-1,3,25-triol is converted to
37
- 26 -
[la,3~,5Z,7E,24S]-24-fluoro-9,10-secocholesta-5,7,10-
(l9)-trien-1,3,25-triol (la,25-dihydroxy-24S~fluorochole-
calciferol), [a]D ^~45 (c = 0.1, CH30H).
B. The starting secocholestane can be preapred in a
manner similar to that described in Example lB starting from
[la,3~]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]pre~n-
5-en-21-oic acid ethyl ester via the following intermedia-
tes:
[la,3~,Z4S]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-
25-(1-ethoxyethoxy)-24-fluorocholest-5-en-21-oic acid
ethyl ester~
[la,3~,24S]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-
25-(1-ethoxyethoxy)-24-fluorocholest-5-en-21-ol,
cla~3~24s]-l~3-bis[(tetrahydro-2H-pyran~2-yl)oxy]
25-(1-ethoxyethoxy)-24-fluorocholest-5-en-21-ol 21-(4-
methylbenzenesulfonate),
[la,3~,24S]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-
25-(1-ethoxyethoxy)-24-fluoro-21-iodocholest-5-ene,
[la,313,24S]-1,3-bis[(tetrahydro-2H-pyran-2-yl)oxy]-
25-(1-ethoxyethoxy)-24-fluorocholest-5-ene,
[la,3~,24S]-24-fluorocholest-5-en-1,3,25-triol,
[la,3~,24S]-24-fluorocholest-5-en-1,3,25-triol 1,3-
diacetate,
[la,3~,7~,24S]-7-bromo-24-fluorocholest-5-en-1,3,25-
triol 1,3-diacetate,
[la,3~,24S]-24-fluorocholesta-5,7-dien-1,3,25-triol
30 1,3-diacetate,
[la,3~,6Z,24S]-24-fluoro-9,10-secocholes-ta-5(10),6,8-
trien-1,3,25-triol 1,3-diacetate.
C. The intermediate, 4-(1-ethoxyethoxy)-3S-fluoro-l-iodo-
35 4-methylpentane, [a]D -38 (c = 1, CHCl3), can be pre-
pared in a manner similar to that described in Example lC
startiny from
3~
(+)-2R-hydroxy-4-butyrolactone via the following
compounds:
2S-fluoro-4-butyrolactone, [a]23 -51 (c = 0.9,
CHC13),
4-methyl-3S-fluoro-1,4-pentanediol, [a~D -40 (c = 1,
CHC13),
4-methyl-3S-fluoro-1,4-pentanediol 1-acetate, [a]D
_49D (c = 0.5, CHC13),
4-(1-ethoxyethoxy)-3S-fluoro-4-methyl-1-pentanol
1-acetate,
4-(1-ethoxyethoxy)-3S-fluoro-4-methyl-1-pentanol,
[a]D -16 (c = 1, CHC13),
4-(1-ethoxyethoxy)-3S-fluoro-4-methyl-1-pentanol
1-t4-methylbenzenesulfonate).
Example 3
Capsules are manufactured by dissolving the ingredient
1, 3 and 4 into ingredient 2 under nitrogen and by
encapsulating.
Ingredients mg/capsule
25 1. la,25-dihydroxy-24~-
fluorocholecalciferol 0.00010 0.00025 0,00050 0.00100
2. polyethylene glycol 400 200.00 200.00 200.00 200.00
3. butylated hyclroxy
anisole 0.100 0.100 0.100 0.100
30 4. ascorbyl palmitate 1.00 1.00 1.00 1.00
Example 4
A solution ~or intramuscular injection is prepared bv
~ 35 dissolving ingredient 1 in 2 under nitrogen.
:::
1. la~25-dihydroxy-24R-
fluorocholecalciferol 0.10 mg 0.50 mg 1.00 mg
2. 95% ethanol - 5~/~ water 2.00 ml 3.00 ml 3.00 ml
