Note: Descriptions are shown in the official language in which they were submitted.
- ~Z~3681~Z
RAN 4212/32
The invention is based on the finding of novel whoosh
logical properties of the C-25 R- and S-epimers of lug-
trihydroxycholecalciferol, hereinafter denominated the R-
and S-epimers. Specifically it has been found that the R- or
S-epimer lower higher than normal levels of endogenously
10 produced la,25-dihydroxycholecalciferol.
Accordingly, the invention relates to the R- or S-
epimer as an agent for the treatment of disease states
characterized by higher than normal serum levels of endow
15 venously produced la,25-dihydroxycholecalciferol or kirk-
terraced by conditions where there is an increased sunsuit-
viny to la,25-dihydroxycholecalciferol. The invention also
relates to the use of the R- or S-epimer for the alone-
mentioned treatment, as well as to pharmaceutical compost-
20 lions on the basis of the R- or S-epimer for this treatment
and to a corresponding method of treatment comprising the
administration of the R- or S epimer.
Specifically included among the above mentioned disease
25 states are hypercalcemia, sarcoidosis, hypercalciuria,
nephrolithiasis and nephrQcalcinosis.
As indicated above, the R- and S-epimer lower the
serum level of la,25-dihydroxycholecalciferol. Additionally,
30 both the R- and S-epimers promote bone mineralization in
vitamin deficient animals, but only the R-epimer promotes
bone mineralization in disodium ethanes 1-hydroxy-1,1
diphosphonate-blocked animals.
The foregoing activities can be demonstrated in the
following tests:
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(a) Anti-rachitogenic activity in chicks
White Leghorn chicks are placed on a vitamin Defoe
client diet containing 1% calcium and 0.7% phosphorus, and
are housed under ultraviolet-fre2 lighting. The test come
pounds are dissolved in propylene glycol and administered
orally for 21 consecutive days to the chicks which are one
to two days of age at the start of treatment. Controls are
treated with vehicle alone. Chicks are autopsies on the day
after the last treatment day and the tibia ash weight is
determined Nine to ten chicks are used for each treatment
group and for the control group. The mean tibia ash weights
expressed in my are given in Table I. The results show
that the R- and S-epimers possess similar antirachitogenic
activity.
Tale I
Mean tibia ash weight (my)
20 Dose
(mg/chick/day) S-epimer R-epimer
0 112.1+6.2
124.9~2.8 122.6+ 4.1
100 159.4+4.4 157.5+ 5.3
25300 187.0+8.7 207.8+ 7.8
1000 210.4+7.8 213.3+10.0
(b) Intestinal gala us absorption in chicks
White Leghorn chicks are placed on the vitamin D-
deficient diet and are housed under ultraviolet-free
lighting for 21 days. A single oral dose of 1 mug of test
compound dissolved in propylene glycol is administered. At
various tires after dosing, 2 Sue of kiwi (chloride) is
35 given orally, and serum radioactivity is measured 45 minutes
after administration of the isotope. Vehicle-treated con-
trots are included at each time period. Ten chicks are used
in each treatment and control group. The results given in
~L~0~38Z
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Table II show that the the S- and R-epimers possess similar
intestinal kiwi absorption activity.
table II
Treatment Time (hours) Serum Cay (cpm/0.2 ml)
Vehicle, 0.2 ml 3 1338~ 51
S-Epimer 1977+122
10 R-Epimer 2077+174
Vehicle, I ml 6 1345+ 89
S-Epimer 2067+128
R-Epimer 1992+212
I
(c) Prevention of EHDP-induced mineralization block in rats
Charles River male rats are treated for 10 consecutive
days with disodium ethanes 1-hydroxy-1,1-diphosphonate (EHDP).
20 The compound is given subcutaneously on each treatment day
at a dose of 2 mg/0.2 ml/rat in distilled water. The test
compounds are administered orally, dissolved in propylene
glycol, on each treatment day. Rats are autopsies on the
day after the last treatment day and the tibias are pro-
25 cussed by silver impregnation of the bone salts. Epiphysealplate widths are measured with a microscope. activity is
based upon dose dependent narrowing of the widened opt-
fuzzily plate induced by EHDP. Ten rats are used in each
treatment group. Positive (EHDP alone) and negative
(vehicle alone) control groups of ten rats each, are inkwell-
dyed in each experiment. The results given in Table III
indicate that the R-epimer caused calcification of the
tibia epiphyseal plate in EHDP-blocked rats while the
S-epimer did not.
~206~8Z
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Table III
Dose Mean tibia epiphyseal plate width
(mcg/rat/day) (micron)
EHDP + EHDP EHDP +
_ S-epimer alone R-epimer
0 1190+42
1162+36 611+51
.
Vehicle controls (no EHDP) 376+13
(d) Effects on the serum levels of Lydia and
ODE
The effect of subcutaneous administration of the S-
and R-epimers on serum levels of la,25-dihydroxycholecal-
ciferol and 25-hydroxycholecalciferol (Lydia and
ODE) was determined according to known methods
(Archives ox Biochemistry and Biophysics, Vol. 201, No. 1,
1980, Z77-285). The results are shown in Table IV. The
effect was initially shown in a 7 day experiment and later
in a 28 day treatment study which demonstrated that the
reduction of Lydia levels was sustained. The ability
25 to control endogenous production of Lydia is glint-
gaily useful in the treatment of among other disease
states, sarcoidosis and hypercalciuria~
Table IV
Daily treatment No. of No. of Serum levels of
daysratsla~25(0H)2D3 ODE
~pg/ml) (ng/ml)
Vehicle, 0.2 ml- 7 1899.6+0.5 37.6+1.9
35 S-Epimer 1 mug 127.9+2.8 32~0+1.7
R-Epimer l mug 1214.0+4.1 36.0+1.8
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-- 5
Vehicle, 0.2 ml pa 10 3~.8+7.3 64.2+4.3
S-Epimer 1 mug 108.0+2.3 40.6+3.4
R-Epimer 1 mug 108.2+1.7 34.7+4.2
The R- and S-epimers may be administered in dosages
that are in the range of 0.5 to 500 micrograms per day. They
are preferably administered orally, but can also be ad mini-
stored subcutaneously, intramuscularly, intravenously or
intraperitoneally, for instance in form of tablets, cap-
10 sulks or elixirs or oral administration; or in sterile
solutions or suspensions for parenteral administration.
About 0.5 to 500 micrograms of the R- or S-epimer is
compounded with pharmaceutically acceptable adjutants,
such as a vehicle, carrier, excipient, binder, preservative,
15 stabilizer and/or flavor in a unit dosage form.
Illustrative of the adjutants which may be incorporated
into tablets or capsules are a binder, such as corn starch
or gelatin; an excipient, such as dicalcium phosphate; a
20 disintegrating agent, such as corn or potato starch or
alginic acid; a lubricant, such as magnesium Stewart; a
sweetening agent, such as sucrose; a flavoring agent, such
as peppermint. Other materials may be present as coatings
or to otherwise modify the physical form of the dosage unit.
25 For instance, tablets may be coated with shellac, sugar or
both. A syrup or elixir may contain the active compound,
sucrose as sweetening agent, methyl and propel parabens as
preservatives, a dye and a flavoring, such as orange flavor.
Sterile compositions for injection can be formulated
according Jo conventional pharmaceutical practice by dozily-
vying or suspending the active substance in a vehicle, such
as water for injection, a naturally-occurring vegetable oil,
such as sesame oil, or a synthetic fatty vehicle, such as
35 ethyl owlet. Buffers, preservatives and antioxidant can
also be incorporated.
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Example 1
Tablet Formulation
mg/tablet
1. The R- or S-epimer of
la,25,26-trihydroxychole-
calciferol 0. 025 0 .100 0 . 5
2. Lactose 157.975 157.900 157.5
3, Microcrystalline cellulose 20.00 20.0
4. Modified starch 20. 000 20. 0
5. Magnesium Stewart 2. 000 2. 0
Total 200.000 mg200.000 my 200. 0 my
The ingredients 1 to 4 are mixed and if necessary
milled. After addition of the magnesium Stewart, the mix-
lure is milled and then pressed to tablets.
Example 2
; 20
- Capsule Formulation
mg/capsule
1, The R- or S-epimer of
25 la,25,26-trihydroxychole-
calci~erol 0.025 0.1000.500
2. Lactose 159. 975 159. 90159 .50
3. Modified starch 20.0 20.0 20.0
4. Talc 20.0 20.0 _20.0
I Tuttle my 200 mg200 my
The ingredient 1 is dissolved in alcohol. The inure-
dints 2 and 3 are mixed and the solution of 1 is spread
over the mixture which is then dried overnight. The mixture
35 is screened, then mixed with talc and filled into capsules.
~LZC36~3~3Z
Example 3
The drug can be dissolved in a pharmaceutically accept
-table solvents, such as alcohol, propylene glyco~, glues-
fine or polyethylene luckily. Surfactants, such as polyethy-
tone glycol, sorbitan esters, ductile sodium sulfosuccinate,
polyoxyethylene-polyoxypropylene co-polymer can also be
added for solubilization of the drug. A preservative can be
added to the formulation for the prevention of microbial
10 growths. Illustrative of capsule formulations are:
a) mg/capsule
The R- or S-epimer of
la,25,26-trihydroxychole-
15 calfiferol oily 0.5Q
Polyethylene glycol (PUG 400.0 400.00
Butylated hydroxy-
anisol (BRA) 0.2 0.2 0.2
Acquirable palpitate lo lo lo
Dissolve BRA and acquirable palpitate in PEG. Add the
R- or S-epimer of la,25,26-trihydroxycholecalciferol and
dissolve under an atmosphere of nitrogen. The liquid is
filled into social capsules.
I
b) capsule
The R- or S-epimer of
la,25,26-trihydroxychole-
calciferol oily 0.50
30 PEG 400 (or PEG 6000) 200.0 200.0 200.0
Polyoxyethylene Sorbitan moo-
owlet or menstruate (Polysorbate
80 or Polysorba~e 60) 200.0200.0 200.0
BRA 0.2 0.2 0.2
35 Acquirable palpitate lo lo lo
~1i206~382
-- 8 --
Warm the mixture of PEG 6000 and Polysorbate 60. Add
to it BRA and acquirable palpitate. Add the R- or S-epimer
of la,25,26-trihydroxycholecalciferol under an atmosphere
of nitrogen. Fill into hard-shell capsules.
c)
The R- or S-epimer of
la,25,26~trihydroxychole-
calciferol 0.025 0.1 0.50
10 PEG 400 100.0 100.0 100.0
PEG 4000 300-0 300 300
BRA 0.1 0.1 0.1
Butylated hydroxytoluene
(BUT) 0.1 0.1 0.1
US Acquirable palMitate1.0 1.0 1.0
Warm a mixture of PEG 400 and PEG 4000. Add BUT, BRA
and acquirable palpitate. Add the R- or S-epimer of lay 25,26-
trihydroxycholecalciferol and dissolve under a stream of
20 nitrogen. Fill into hard-shell capsules.
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