Note: Descriptions are shown in the official language in which they were submitted.
TN-7633
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DESCRIPTION
TITLE OF THE INVENTION
Pharmaceutical Preparation for Chronic Hepatitis
Treatment
TECHNICAL FIELD
The present invention relates to a pharmaceutical
preparation for chronic hepatitis treatment.
BACKGROUND ART
Chronic hepatitis is an inflammatory disease of the
liver which persists for at least 6 months and is
Lo considered to be caused by an infection by the hepatitis
virus, autoimmunity, certain kinds of drugs, and the
like. Generally, it has been found that a liver disease
will take the course of an occurrence of a transitional
extension, chronic change, and inverterate hepatitis,
and in some cases, that the disease takes 10 or more
years before healing or the disease may be migrated to
cirrhosis, and a further complication of primary
hepatoma may cause death in many cases. The migration
percentage from chronic hepatitis to cirrhosis is not
constant, depending on the source, but is approximately
40%. On the other hand, the healing percentage is much -
smaller, and has been estimated to be around 10%.
Accordingly, about half of the cases have persistent
subjective symptoms and liver function abnormalities and -
25 are difficult to cure. ~~
Chronic hepatitis is classified into an active type
and a nonactive type, by liver function and liver
biopsis image, and the active type and nonactive type
are mutually migratable to each other. The therapy -
therefore is performed at the time when an active lesion
exists and the practiced drug therap~ is based on
instructions on food intake and how to relax.
In the prior art, as a drug therapy, a therapy with
the use of strong Neo Minofagen C ~ as liver protecting
agents, interferon (IFN), arabinoside-A (Ara-A) as
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antiviral agents, and glucocorticoid (GC) withdrawal as
the immune control therapy have been practiced, but some
problems arise in the use of these prior art methods.
Namely, in the case of the Strong Neo Minofagen C ~,
since it is injected, it is not suitable for a long term
therapy; in the case of IFN, fervescence, a reduction of
leukocyte and platelet numbers, and other side effects
occur; in the case of Ara-A, a reduction in leukocyte
and platelet numbers and a peripheral nervous disorder
1~ are liable to be exhibited; and in the case of GC, since
the DNAp activity is inevitably increased one week after
administration, it must be carefully administered.
Accordingly, there is an urgent need to develop a
therapeutic composition which can be orally adminis-
tered, has few side effects and has a high efficacy.
DISCLOSURE OF THE INVENTION
Accordingly, the objects of the present inventionare to eliminate the above-mentioned disadvantages of
the prior art and to provide a pharmaceutical
preparation for chronic hepatitis treatment capable of
being orally administered and having few side effects
and a high efficacy.
Other objects and advantages of the present
invention will be apparent from the following
description.
In accordance with the present invention, there is
provided a pharmaceutical preparation for chronic
hepatitis treatment comprising an active vitamin D, as
an active ingredient, and a carrier therefore.
BRIEF DESCRIPTION OF THE DRAWINGS
The present invention will be better understood
from the description set forth below with reference to
the accompanying drawings; wherein;
Figure 1 shows the change of GOT when l~-hydroxy-
vitamin D3 is administered; and
Fig. 2 shows the change of GPT when l~-hydroxy-
vitamin D3 is administered.
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BEST MODE OF CARRYING OUT THE INVENTION
Active vitamin D' s such as 1~-hydroxyvitamin D, 1
24(R)-dihydroxyvitamin D, 1~ 25-dihydroxyvitamin D,
promote an absorption of calcium in the small intestine,
promote bone resorption and bone formation in bones, and
are well known in the art as a therapeutic agent for
diseases due to various calcium metabolism abnor-
malities.
The present inventors made an intensive investiga-
tion of the effects of the active vitamin D on chronichepatitis, and consequently, surprisingly found that the
active vitamin D has the action of lowering the GOT and
GPT, which are indices of liver function.
The active vitamin D usable in the present
invention includes the active vitamin D2 ~ the active
vitamin D3 , and derivatives thereof. Typical examples
of such compounds are l~-hydroxyvitamin D, 1~ 24(R)-
dihydroxyvitamin D, 1~, 25-dihydroxyvitamin D, 1~ 24
25-trihydroxyvitamin D, 24~ 24-difluoro-1~ 25-dihy-
droxyvitamin D, 26, 26, 26~ 27, 27, 27-hexafluoro-1~,
25-dihydroxyvitamin D, 25-hydroxyvitamin D3-26, -
23-lactone and 1~ 25-dihydroxyvitamin D3-26, - :
23-lactone. Among these examples, 1~-hydroxy-
vitamin D3 , 1~ 24(R)-dihydroxyvitamin D3 , 1~
25 25-dihydroxyvitamin D3 are preferable, and particularly, -
1~, hydroxyvitamin D3 is most preferred.
These active ingredients can be used as oral
preparation in the form of, for example, softt capsules,
hard capsules, tablets, powders, granules, or syrup, or
optionally can be used as parenteral preparations in the
form of injections, or external preparations, with the
use of appropriate vehicles including excipients
according to conventional methods.
Examples of the vehicles usable in the present
invention are vegetable oil or mineral oils, white
petrolatum, branched chain fats or oils, animal fats and
high molecular weight alcohols for liquid agents or
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external application agents. Among these vehicle~,
vegetable oils (corn oil, cotton oil, coconut oil,
almond oil), especially middle chain length fatty acid
triglycerides are preferred.
Examples of the vehicles for solid agents usable
are cellulose derivatives (crystalline cellulose,
hydroxypropyl cellulose, hydroxypropyl-methyl cellulose,
methyl cellulose), polyvinylpyrolidone, dextrin,
cyclodextrin, casein, lactose, mannitol, gelatin or
starch. Among them, lactose, polyvinylpyrolidone etc.
are preferred. Although there are no limitations to the
content of the active ingredient, i.e., the active
vitamin D in the composition, the content of the active
ingredient is preferably (0.00004 - 0.2) x 10 4% by
weight, more preferably (0.001 - 0.08) x 10 4~ by weight
in the composition.
The dose of the active ingredient is preferably
about 0.01 to 10 ~g/day/person, more preferably 0.25 to
4.0 ~g/day/person, and the administration is preferably
made 1 to 3 times/day. Preferably, preparations are
obtained which satisfy such conditions.
The method of the present invention can be also
used in combination with known drug therapy therapeutic
agents.
EXAMPLES
The present invention will now be further illus-
trated by, but is by no means limited to, the following
Examples.
Exam~le 1
Active vitamin D3 , l~-Hydroxyvitamin D3 ,
(l~-OH-D3) was orally administered to 11 patients with
chronic hepatitis confirmed by liver biopsy and
serologic study (9 males, 2 females, average age 51.5)
at daily doses of 0.5 to 2.0 ~g. Soft capsules
comprising 0.5 - 2.0 ~g of 1~-OH-D3 and 150 mg of
triglycerides of medium-chain fatty acids were used for
the administration. The liver function was evaluated by
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serologic study (GOT, GPT) once a month for 6 months
before and after the administration.
The mean GOT and GPT levels for 6 months before and
after the administration are shown in Fig. 1 and Fig. 2,
respectively. It is clear from Fig. 1, that the mean
GOT level was reduced from 173.9 KU to 83.1 XU by the
treatment. It was also confirmed that the mean GPT
level was reduced from 172.4 KU to 83.2 KU. These
reductions of GOT and GPT levels to 100 or less after
administration, it is expected that the migration from
chronic hepatitis to cirrhosis can be inhibited. Also,
hypercalcemia and side effects were not observed during
the test period.
Example 2: Anti-BLP AntibodY induced HePatitis
Model
The experiment was performed according to the
method of Nagai et al. (Japanese Journal of Inflammation
6, 361 (1986)). Namely, liver was removed from DBA/2
strain male mice (male, 7 weeks old), and a 50 wt%
homogenate was prepared with physiological saline.
Supernatant fluid obtained by centrifugation at 4C and
8000 rpm for 30 minutes was adjusted to pH 4.8 with
acetic acid. After centrifugation, saturated ammonium
sulfate was added to the supernatant and the fractions
precipitated at 35 to 60% were obtained. The
precipitates were dissolved in distilled water, and
dialyzed against 0.005 M Tris-HCl buffer (pH 8.0). The
dialyzed solution was applied to a DEAE cellulose column
chromatography equilibrated with 0.005 M Tris-HCl buffer
(pH 8.0), and the fractions eluted were made into basic
liver protein (BLP).
One ml of BLP (300 ~g protein/ml) was emulsified -
with an equal volume of Complete Freund's Adjuvant
(CPA), the emulsified mixture was immunized to a New
Zealand White rabbit (female, 2.0 to 2.5 kg) every week
for 4 to 6 weeks. Ten days after the final -
immunization, blood was collected from the cervical
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aorta and serum was obtained. After inactivation of
complement by heat treatment at 56C for 30 minutes, the
serum was absorbed with mouse kidney homogenate and rat
erythrocytes to obtain a specific antibody to BLP.
To DBA/2 strain male mice (7 weeks old) were
intraperitoneally injected with 0.25 ml of rabbit
7 globulin (RGG~ 4 mg/ml) emulsified with an equal
volume of CFA. Five days later, 0.6 ml of the anti-BLP
antibody was injected through the tail vein, and blood
for GPT assay was collected from the right ventricle
18 hours after antibody in~ection.
l~-OH-D3 (0.8, 4, or 20 ng/kg/day) was orally
administered once a daily for 7 days from one day before
the RGG injection.
The results are shown in Table 1.
Table 1
Sample n GPT (KU/~)
Control 8 72.5 + 19.0
l~-OH~D3 0.8 ng/kg 8 43.3 + 7.5
l~-OH-D3 4 ng/kg 8 39.4 + 13.0
1~-OH-D3 20 ng/kg 8 40.0 + 14.9
RGG 6 11.8 + 3.0
Normal 4 14.0 + 0.4
In anti-BLP antibody-treated mice, the mean GPT
level was 72.5 KU/l, which was clearly higher than that
of normal mice and RGG-treated mice: mean levels 14.0
and 11.8 KU/l, respectively. In contrast, l~-OH-D3 , at
doses of 0.8, 4 and 20 ng/kg/day, inhibited the mean GPT
level by 40.3, 45.7, and 44.8%, respectively.
ExamPle 3: P. acnes LPS-induced HePatitis Model
Balb/c mice, (male, 8 weeks old) were injected with
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0.1 mg of heat-killed Propionibacterium acnes (P. acnes)
through the tail vein. Seven days later, 3 ~g of
lipopolysaccharide (LPS) derived from E. coli was
injected through the same route. Blood was collected
18 hours thereafter, from the right ventricle, and the
serum GPT level was measured.
l~-OH-D3 (4, 100 ng/kg/day) was orally administered
for 9 days from one day before the P. acnes treatment.
The results are shown in Table 2.
Table 2
Sample n GPT (KU/~)
Control 10 680.0 + 178.6
l~-OH-D3 0.004 ~g/kg 8 495.0 + 68.6
1~-OH-D3 0.1 ~g/kg 8 321.3 + 79.8
Normal 5 37.2 + 4.0
In P. acnes and LPS-treated a hepatitic mice, the
mean GPT level was increased to 680 XU/l. In contrast,
1~-OH-D3 , at doses of, 4, and 100 ng/kg/day inhibited
the mean GPT level by 27.2~ and 52.8~, respectively.
Exam~le 4
Active vitamin D3 , 1~-hydroxyvitamin D3 , was
orally administered, at a dose of 1.5 ~g/day for -
14 months into to a patient (female, age 50) with
chronic hepatitis confirmed by liver biopsy and
serologic study. The liver function (GOT, GPT) and the
liver biopsy were evaluated.
The GOT and GPT levels were reduced from 130 KU and ~ -
132 KU, and from 132 KU to 30 KU, respectively.
It was confirmed by a tissue observation of the
liver biomass examination that the remarkable decrease
in the inflammation observed before and after the
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administration. The histopathological finding was not
substantially different from normal tissue. No side
effects such as hypercalcemia were observed during the
examination.
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