Note: Descriptions are shown in the official language in which they were submitted.
~lV~
RAN 4212/35K
The inventlon is based on the finding of novel phy-
siological properties of 24,24-difluoro la,Z5-dihydroxy-
cholecalciferol, hereinafter denominated the fluorinated
compound. Specifically it has been found that the fluori-
nated compound is active against osteoporosis and in pre-
10 venting milk fever, and can thus be utilized in the treat-
ment of osteoporosis by administering to a patient suffe-
ring from osteoporosis an effectlve amount of the fluori-
nated compound, or can be utilized in the prevention of
milk fever by administering to a preparturient female
15 ruminant an effective amount of the fluorinated compound.
Accordingly, the invention relates to the fluorinated
compound as an agent for the treatment of osteoporosis or
the preventive treatment of milk fever. The invention also
20 relates to the use of the fluorinated compound for the
aforementioned treatments as well as to pharmaceutical
compositions on the basis of the fluorinated compound for
these treatments and to corresponding methods of treatment
comprising the administration of the fluorinated compound.
The foregoing activities of the fluorinated compound
can be demonstrated in the following tests:
a) Anti-rach~ Lenic activitv in chfi~OEks
White Leghorn chicks are placed on a vitamin D-deficient
diet containing 1% calcium and 0.7% phosphorous, and are
housed under ultraviolet-free lighting. The test compound
is dissolved in propylene glycol and administered orally
35 for 21 consecutive days to the chicks which are one to two
days of age at Ihe start of treatment. Controls are treated
Mé/ 22.3.83
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with vehicle alone. The chicks are autopsied on the day
after the last treatment day and the tibia ash weights are
determined. Nine to ten chicks are used for each treatment
group and for the control group. The mean tibia ash weights
expressed in mg are given in Table I. The results show
that the fluorinated compound possess potent anti-rachito-
genic activity.
Table I
~ose
(ng/chick/day) Mean tibla ash weight (mq)
0 120.7+5.9
1 111.7+5.8
3 151.5+4.7
227.1+8.2
244.8+7.4
20 b) Intestinal calcium absorPtion in chicks
White Leghorn chicks are placed on a vitamin D-defi-
cient diet and are housed under ultraviolet-free lighting
for 21 days. A single oral dose of test compound dissolved
25 in propylene glycol is administered. At various times
after dosing, 2 ~Ci of 45~a (chloride) are given orally
and serum radioactivity is measured 45 minutes after admini-
stration of the isotope. Vehicle-treated controls are
included at each time period. Ten or eleven chicks are used
30 in each treatment and control group. The results given in
Table II for la,25-dihydroxycholecalciferol (compound A)
and for the fluorinated compound (B), show that the latter
has potent intestinal calcium absorption activity of long
duration (96 hours after a single oral 100 nanogram dose)
35 and consequently possesses utility in the prevention of
milk fever in preparturient female ruminants.
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Table II
Treatment Time (hours) Number o Serum Ca
chicks(cp~/0.2 ml)
Vehicle 0.2 ml 24 11 992 + 81
Compound A 100 ng 11 1800 + 181
Compound B 100 ng 11 2064 + 170
10 Vehicle 0.2 ml 48 11 769 + 90
Compound A 10~ ng 11 1006 + 133
Compound B 100 ng 11 1539 + 99
Vehicle 0.2 ml 72 10 647 + 69
15 Compound A 200 ng 11 710 + 62
Compound ~ 100 ng 11 1164 + 90
Vehicle 0.2 ml 96 10 586 + 70
Compound B 100 ng 10 998 + 61
c) Prevention of EHDP-induced mineralization block in rats
Charles River male rats are treated for 10 consecuti~e
days ~ith the disodium salt of ethane-1-hydroxy~ diphos-
25 phonate (EHDP). This compound is given subcutaneously oneach treatment day at a dose of 2 mg/0.2 ml/rat in distilled
water. The fluorinated compound is administered orally
dissolved in propylene glycol on each treatment day. The
rats are autopsied on the day after the last treatment day
30 and the tibias are processed by silver impregnation of the
bone salts. The epiphyseal plate widths are measured with
a microscope. Activity i5 based upon dose-dependent narro-
wing of the widened epiphyseal plate induced by EHDP. Ten
rats are used in each treatment group. Positive (EHDP alone)
35 and negative (vehicles alone) control groupsof ten rats
each are included in each experiment. The results given in
Table III show that the fluorinated compound calcified the
tibial epiphyseal plate in EHDP-blocked rats.
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Table III
Dose Mean tikial epiphyseal plate width
(ng/rat/day) (micron)
0 1182 + 51
1 839 + 18
3 674 + 18
540 + 16
1030 412 + 9
Vehicle controls
(no EHDP) 440 + 2
The fluorinated compound can be administered in
15 dosages that are in the range of about 50-200, preferably
100 nanograms per day for the treatment of osteoporosis.
It can be administered orally, subcutaneously, intramus-
cularly, intravenous~y or intraperitoneally, for instance
as tablets, capsules or elixirs for oral administration,
20 or in sterile solutions or suspensions for parenteral
administratiQn. About 50-200 nanograms of the fluorinated
compound is compounded with a pharmaceutic~lly acceptable
vehicle, carrier, excipient, binder, preservative, stabili-
zer and/or flavor in a unit dosage form. Illustrative of
25 the adjuvants which may be incorporated into capsules are
a binder, such as corn starch or gelatin; an excipient,
such as dicalcium phosphate; a disintegrating, such as
corn or potato starch or algenic acid; a lubricant, such
as magnesium stearate; a sweetening agent, such as sucrose;
30 a flavoring agent, such as peppermint. other materials may
be present as coatings or to otherwise 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,
35 methyl and propyl parabens as preservatives, a dye and a
flavoring, such as orange flavor. Sterile compositions for
injectlon can be formulated according to conventional
pharmaceutical practice by dissolving or suspending the
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fluorina~ed compound in a vehicle, such as water for lnjec-
tion, a naturally-occurring vegetable oil, ~uch as sesame
oil, or a synthetic fatty vehicle, such as ethyl oleate.
Buffers, preservatives and antioxidants can also be incor-
porated.
The fluorinated compound can be administered indosages in the range of about 30-500 micrograms per day
for the prevention of milk fever in pregnant ruminant ani-
10 mals, especially in pregnant female bovines, preferablyabout one to four days prior to parturation. The fluorinated
compound can be formulated for oral administration by in-
corporation of 30-500 micrograms into fatty acid pellets.
Sterile compositions for injection and/or topical admini-
15 stration can be formulated according to conventionalpractice by dissolving or suspending the fluorinated com-
pound in a vehicle, such as a 5-10% ethanol-water mixture;
a naturally-occurring ve~etable oil, such as sesame oil;
or a synthetic fatty vehicle, such as ethyl oleate. For
2~ example, a suitable formulation for intravenous injection
would be 2-3 ml of a 5-10% ethanol-water solution containing
30-500, preferably 30-400 micrograms of the fluorinated
compound. Exemplary of a suitable formulation for topical
administration would be a vegetable oil solution or sus-
25 pension containing 30-500, preferably 250-50Q micrograms of
the fluorinated compound. The fluorinated compound may also
be formulated for intramuscular injection by suspension of
50-500 micrograms of the fluorinated compound in a vehicle
such as a 5-10% ethanol-water mixture or a 5-10% propylene
30 glycol-water mixture. Buffers, preservatives and/or anti-
oxidants can be incorporated into the foregoing formula-
tions as required.
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Example
A capsule for the treatment of osteoporosls con-
taining pro capsule:
1. 24,24-difluoro-la,25-
dihydroxycholecalciferol 50 ng 100 ng 200 ng
2. polyethylene glycol 400 200 mg 200 mg 200 mg
3. butylated hydroxyanisole 0.1 mg 0.1 mg 0.1 mg
10 4. ascorbyl palmitate1.0 mg 1.0 mg 1.0 mg
may be obtained by dissolving items 1, 3 and 4 in
item 2 under an atmophere of nitrogen and encapsulating.
