Note: Descriptions are shown in the official language in which they were submitted.
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SPECIFICATION
QUINONE-BASED THERAPEUTIC AGENT FOR HEPATOPATHY
Technical Field
The present invention relates to an agent for treating hepatic
disease, and more specifically to an agent for improving hepatic
cancer prognosis, containing menatetrenoneas an activeingredient.
Background Art
It is known that there is a high rate of portal venous invasion
(hereinafter referred to as 'PVI') among hepatocellular carcinoma
(hereinafter referred to as 'HCC' ) patients, and once PVI has occurred
the prognosis is very poor. It is known that a high des-y-carboxy
prothrombin (hereinafter referred to as 'DCP' ) level in HCC patients
is closely linked to subsequent development of PVI (see Koike Y.,
Cancer 2001, 91 : 561-9) . DCP is also called PIVKA-II (protein induced
by vitamin K absence or antagonist II ) . DCP is a pro thrombin that
does not have normal coagulation activity, and is known to increase
in level in the case of vitamin K (hereinafter referred to as 'VK' )
deficiency: DCP is thus a protein that is used as a marker for VK
deficiency or impaired VK absorption.
Moreover, it has been reported that if VK is administered to
HCC patients with a high DCP level, then the serum DCP level drops
(see Cancer 1992, 69:31-8), and that upon administering vitamin
K-II (hereinafter referred to as 'VK-II') to a DCP-producing HCC
cell line in Vitro, cell proliferation is inhibited (see Hepatology
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1995, 22:876-82).
However, until now no clinical data had been collected with
regard to it being possible, by administering VK-II to patients
after HCC treatment, to inhibit the occurrence of PVI, and to inhibit
the recurrence of hepatocellular carcinoma, thus improving the
prognosis.
In view of the above, it is an obj ect of the present invention
to provide an excellent agent for treating or preventing hepatic
disease.
Disclosure of Invention
The present inventors accomplished the present invention by
being the first to discover that administering an oral VK-II
preparation to DCP-producing HCC patientscontributesto inhibiting
the occurrence of PVI after HCC treatment and improving the prognosis,
and moreover inhibits the recurrence of hepatic cancer after
treatment.
The above object is attained through an agent for treating
or preventing hepatic disease containing menatetrenone as an active
ingredient thereof.
According to a preferable aspect of the present invention,
in the case of the above agent, the hepatic disease is hepatic cancer.
According to a preferable aspect of the present invention,
in the case of the above agent, the hepatic cancer is des-y-carboxy
prothrombin (DCP) positive hepatic cancer.
According to a preferable aspect of the present invention,
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the above agent improves prognosis after hepatic cancer treatment.
According to a preferable aspect of the present invention,
the above agent is an agent that inhibits occurrence of portal venous
invasion (PVI).
Moreover, the above obj ect is attained through an agent for
inhibiting occurrence of portal venous invasion (PVI) containing
menatetrenone as an active ingredient thereof.
Moreover, the above obj ect is attained through an agent for
improving survival rate after hepatic cancer treatment containing
menatetrenone as an active ingredient thereof.
Moreover, the above object is attained through an agent for
inhibiting recurrence of hepatocellular carcinoma containing
menatetrenone as an active ingredient thereof.
Moreover, the above object is attained through an agent for
reducing DCP level containing menatetrenone as an active ingredient
thereof.
Moreover, the above object is attained through a method of
preventing portal venous invasion (PVI), comprising administering
to a patient an effective dose of a medicine containing menatetrenone
as an active ingredient thereof.
Moreover, the above object is attained through a method of
inhibiting recurrence of hepatocellular carcinoma, comprising
administering to a patient an effective dose of a medicine containing
menatetrenone as an active ingredient thereof.
Moreover, the above object is attained through a method of
regulating the level of DCP in the blood of a patient, comprising
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administering to the patient an effective dose of a medicine
containing menatetrenone as an active ingredient thereof.
Moreover, the above object is attained through a use of
menatetrenone for producing an agent for inhibiting occurrence of
PVI.
Moreover, the above object is attained through a use of
menatetrenone for inhibiting recurrence of hepatocellular
carcinoma.
Furthermore, the above object is attained through an agent
for treating or preventing hepatic disease containing a vitamin
K as an active ingredient thereof.
The menatetrenone-containing agent for treating hepatic
disease according to the present invention has an excellent effect
of inhibiting occurrence of PVI with hepatic disease, in particular
DCP positive hepatic cancer, and moreover has an excellent effect
of improving the prognosis after hepatic cancer treatment.
Furthermore, the menatetrenone-containing agent for treating
hepatic disease according to the present invention is very useful
in inhibiting recurrence of hepatic cancer after treatment.
Brief Description of Drawings
FIG. 1 shows a selection flowchart of the patient;
FIG. 2 shows a graph indicating changes in DCP level in serum;
FIG. 3 shows a graph indicating changes in PVI incidence rate;
FIG. 4 shows a graph indicating changes in survival rate;
FIG. 5 shows graph indicating the effect of VK-II
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administration on hepatic cancer recurrence inhibition (500
recurrence);
FIG. 6 shows a graph indicating results for HCV cases only
in the trial to verify the effect of VK-II administration on hepatic
cancer recurrence inhibition (50o recurrence);
FIG. 7 shows a graph indicating results in the case of excluding
cases of local recurrence in the trial to verify the effect of VK-II
administration on hepatic cancer recurrence inhibition (500
recurrence);
FIG. 8 shows a graph indicating results in the case of excluding
cases of recurrence within 6 months in the trial to verify the effect
of VK-II administration on hepatic cancer recurrence inhibition
(50o recurrence);
FIG. 9 shows a diagram indicating results of analyzing the
risk ratio (RR) for recurrence using a Cox proportional hazardmodel;
and
FIG. 10 shows graphs indicating results of analyzing the DCP
level before treatment and upon recurrence.
Best Mode for Carrying Out the Invention
Following is a more detailed description of the present
invention by way of examples; however, the present invention is
not limited to these examples.
Hepatic cancer occurs with a high rate from chronic hepatitis
and cirrhosis, which are targeted by the present invention, and
having occurred hepatic cancer reoccurs after treatment with a high
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rate. For example, there are cases of cirrhosis developing from
type C hepatitis or type B hepatitis, and there being recurrence
after the tumors have been excised. According to the agent for
treating hepatic disease of the present invention, the prognosis
after such hepatic cancer treatment can be improved very effectively
(i.e. recurrence can be prevented or treated). Moreover, the
occurrence of PVI, which is one form of recurrence of hepatic cancer
with poor prognosis, can be inhibited very effectively.
The menatetrenone used in the present invention has the
chemical name 2-methyl-3-tetraprenyl-1,4-naphthoquinone, and has
a structural formula as shown below.
O
CH3
CH3
O CH3 CH3 CH3 CH3
Menatetrenone is a yellow crystalline or oily substance, has
no taste or odor, and is readily decomposed by light. Moreover,
menatetrenone hardly dissolves in water. Menatetrenone is also
called vitamin K-II (VK-II), and regarding the pharmacological
action thereof, in the process of protein synthesis of blood
coagulation factors (prothrombin, VII, IX, X), menatetrenone
participates in the carboxylation reaction when glutamic acid
residues are converted into physiologically active
y-carboxyglutamic acid, and menatetrenonepromotes synthesis in the
liver of normal prothrombin and so on, and activates the hemostasis
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mechanism in a living body, thus physiologically realizing
hemostasis.
The menatetrenone that is the active ingredient in the medicine
according to the present invention may be in the form of an anhydride
or a hydrate. Moreover, menatetrenone has crystal polymorphs, but
there is no limitation, with it being possible for the menatetrenone
to be in any one of the crystal forms, or a mixture thereof.
Furthermore, a metabolite produced through the menatetrenone
according to the present invention being decomposed in a living
body is also included in the scope of the claims of the present
invention.
The menatetrenone used in the present invention can be produced
using a publicly known method, and as a representative example,
can easily be produced using the method disclosed in Japanese Patent
Application Laid-open No. 49-55650; alternatively, the
menatetrenone can easily be procured from a chemical manufacturer.
Moreover, the menatetrenone can be procured as a pharmaceutical
preparation such as a capsule or an inj ection . Regarding the medicine
of the present invention, the menatetrenone may be used as is, or
may be made into a pharmaceutical preparation using a commonly used
method by blending with ingredients that are commonly used as raw
materials of medicinal preparations such as publicly known
pharmaceutically permissible carriers and so on (e. g. excipients,
binders, disintegrators, lubricants, colorants, flavorings,
stabilizers, emulsifiers, absorption promoters, surfactants, pH
regulators, preservatives, antioxidants, etc.). Moreover, other
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ingredients such as vitamins and amino acids may be blended in as
required. Examples of the form of the pharmaceutical preparation
are tablets, a powder, granules, capsules, a syrup, suppositories,
an injection, an ointment, a poultice, and so on.
Moreover, in the present invention, there are no particular
limitations on the form of administration of the menatetrenone,
although oraladministrationispreferable. Menatetrenonecapsules
can be procured as Kaytwo capsules (proprietary name, made by Eisai
Co., Ltd.) and Glakay capsules (proprietary name, made by Eisai
Co . , Ltd. ) , a menatetrenone syrup can be procured as Kaytwo syrup
(proprietary name, made by Eisai Co., Ltd.), and an injection can
be procured as Kaytwo N (proprietary name, made by Eisai Co., Ltd. ) .
The menatetrenone-containing medicine according to the
present invention is useful for treating or preventing hepatic
disease. A preferable dose of the menatetrenone is generally 10
to 200 mg/day, more preferably 30 to 135 mg/day.
Examples
Trial examples of the present invention are given below;
however, these trial examples are merely illustrative, and the
present invention is not limited thereto. A person skilled in the
art could implement the present invention not only through the trial
examples shown below, but also with any of various modifications
within the scope of the claims appended to the specification of
the present application, and such modifications are deemed to be
included in the scope of the claims of the present application.
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Trial Example 1
A clinical trial (randomized prospective controlled study)
was carried out as follows.
Out of patients with hepatocellular carcinoma, ones having
a serum DCP level above 60 IU/L (DCP positive hepatic cancer) were
included in the trial. On the other hand, patients having portal
venous invasion, and patients in which there was alreadyVKmetabolism
through administration of VK or an anti-VK agent, were excluded
from the trial. The details of the trial were as shown in Table
1 .
Table 1
Trial subjects
Subjects included
1. Hepatic cancer patients
2. Serum DCP level >_ 60 IU/L
Subjects excluded
1. Portal venous invasion
2. Cancer metastasize out of liver
3. uncontrolled ascites
4. Bilirubin > 3.0 mg/dl
5. Taking vitamin K preparation, warfarin
Group administered VK-II
Took 45 mg of vitamin K-II (Glakay) three times after
hepatic cancer treatment
Group not administered VK-II
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Hepatic cancer treatment only
Judgement criteria
1. Occurrence of portal venous invasion
2. Death
FIG. 1 shows a selection flowchart of the patient. 126 hepatic
cancer patients were given treatment from February 1999 to November
2001. As the hepatic cancer treatment, percutaneous cauterization
therapy (RFA and/or PEIT) for HCC, treatment via the blood vessels
(TAE or TAI) , or surgical excision was carried out. Of the patients,
5 were excluded from the present trial.
Next, the remaining 121 patients were randomly divided into
a treated group (n = 60 ) and an untreated group (n = 61 ) . The treated
group were orally administered 45 mg/day of VK-II (proprietary name
Glakay, made by Eisai Co., Ltd. ) after the hepatic cancer treatment,
while the untreated group were not administered VK-II.
Follow-up tests were carried out after the hepatic cancer
treatment. As the follow-up tests, ultrasonography (abdominal
echography) was carried out every 3 months, a CT scan was carried
out every 6 months, and the levels of the tumor markers
alpha-fetoprotein and DCP were measured every one month.
Table 2 shows the profile of the patients. No important
differences were found in any of the clinical parameters between
the treated group and the untreated group.
Table 2
Patients Profile
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Treated group Untreated group P
(n = 60) (n=61)
Age 66.97.0 67.37.5 .8
Sex (male/female) 36/24 45/16 .12
Virus (HCV/non HCV) 50/10 52/9 .81
Tumor size (mm) 3211 3518 .27
Number of tumors 4.03.2 4.33.5 .66
Child class (A/B or C) 18/42 27/34 .13
Albumin (g/dl) 3.40.5 3.50.5 .3
Bilirubin (mg/dl) 1.20.7 1.10.9 .4
ALT (IU/L) 5538 6147 .47
Prothrombin (o) 7816 7814 .99
Blood platelets (10q/mm3)10. 86. 0 11.56. 6 .
52
AFP (ng/L) 26687666 15397036 .42
DCP (IU/L) 9852639 11785108 .80
PTA with/without 48/12 41/20 .15
average ~ SD (Median)
FIG. 2 shows a graph indicating changes in the DCP level in
the serum. The solid line is for the treated group, and the dashed
line is for the untreated group. For both the treated group and
the untreated group, the DCP level dropped after the hepatic cancer
treatment. Subsequently, the DCP level remained approximately
constant for 12 months for the treated group, whereas the DCP level
gradually increased for the untreated group.
FIG. 3 shows a graph indicating changes in the PVI incidence
rate. As shown in FIG. 3, for the treated group, the PVI incidence
rate was 2 o after 1 year, and 23 o after 2 years . On the other hand,
for the untreated group, the PVI incidence rate was 23o after 1
year, and 47o after 2 years (P = 0.018).
FIG. 4 shows a graph indicating changes in the survival rate.
As shown in FIG. 4, the survival rate after 2 years was 66~ for
the treated group, but 28o for the untreated group (P = 0.044).
Statistical analysis was carried out on the PVI incidence rate
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and the survival rate for the two groups. That is, calculations
were carried out using the Cox proportional hazard model, and a
test was carried out using the log-rank method. The average
observation period was 12 ~ 8 months.
From the above results, it is suggested that by orally
administering a VK-II preparation, incidence of PVI in DCP positive
HCC patients is suppressed very effectively, and moreover survival
rate is greatly increased, and hence the prognosis after hepatic
cancer treatment is markedly improved.
Trial Example 2
A trial was carried out as follows with an objective of
investigating the effect and safety of VK-II in inhibiting recurrence
of hepatocellular carcinoma after treatment.
61 cases in which hepatocellular carcinoma was diagnosed and
then after treatment therefor necrosis (or curative excision) was
judged by contrast CT to have completely occurred were entered from
March 1999 to March 2001, the entered cases were divided into two
groups, namely a VK-II-administered group for which the end of the
patient ID number was an odd number and a non-administered group
(control group) for which the end of the patient ID number was an
even number, and a VK-II preparation (proprietary name Glakay, made
by Eisai Co., Ltd.) was orally administered at a dose of 45 mg/day
to the administered group. Contrast CT or MRI was carried out every
3 months, and a statistical analysis of the time period until
recurrence was carried out. Specifically, comparison was carried
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out using the Kaplan-Meier method (log-rank test), and the risk
ratio for recurrence was analyzed using the Cox proportional hazard
model.
As shown in Table 3, the average observation period for the
61 entered cases (32 cases in the administered group, 29 cases in
the non-administered group) was 19.6 months (7-32).
Table 3
Administered Control
Subjects group(32 cases) group(29 cases)
Age 63.37.5(48-75) 64.56.7(45-74)
Sex (male/female) 23/9 18/11
Cause of disease 28/3/1 26/2/1
(type C/type B/type B+C)
History of alcohol consumption 10/22 3/26
(addict/non-addict)
First occurrence/recurrence 15/17 14/15
Tumor size (mm) 17.75.1(10-30) 19.46.9(10-38)
Number of tumors 1.500.88(1-4) 1.480.74(1-3)
Log AFP (ng/ml) 1. 470.61 1. 720.91
(0.60-3.09) (0.48-3.88)
PIVKA-II (mAU/ml) 41.865.4 70.3104.1
(8-346) (7-417)
Liver function (LD A/B/C) 15/16/1 13/15/1
Treatment method 1/31 3/26
(excision/non-excision)
Average observation period 24.37.1(13-37) 24.28.3(12-37)
(months)
Upon calculating cumulative recurrence rates, the 1-year
recurrence rate was found to be 12.50 for the VK-II-administered
group and 55.20 for the control group, and the 2-year recurrence
rate was found to be 39.60 for the VK-II-administered group and
85 . 5 o for the control group . From these results, it was found that
the hepatic cancer cumulative recurrence rate was statistically
significantly suppressedfor the VK-II-administered group compared
with the control group.
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FIG. 5 shows a graph indicating the effect of VK-II
administration on hepatic cancer recurrence inhibition (500
recurrence ) . As shown in FIG . 5, the time period until 50 o recurrence
was 26 months for the VK-II-administered group, but was 10 months
for the control group.
Moreover, upon calculating the hepatic cancer cumulative
recurrence rates considering only HCV cases ( type C hepatitis cases ) ,
the 1-year recurrence rate was found to be 7.1o for the
VK-II-administered group and 61 . 5 o for the control group, and the
2-year recurrence rate was found to be 37.80 for the
VK-II-administered group and87.2%for thecontrolgroup. From these
results, it was found that, even when considering only HCV cases,
the hepatic cancer cumulative recurrence rate was statistically
significantlysuppressedfor the VK-II-administered group compared
with the control group.
FIG. 6 shows a graph indicating the results for the HCV cases
only in the trial to verify the effect of VK-II administration on
hepatic cancer recurrence inhibition (50o recurrence). As shown
in FIG. 6, the time period until 50o recurrence was 26 months for
the VK-II-administered group, but was 10 months for the control
group.
FIG. 9 shows a diagram indicating the results of analyzing
the risk ratio (RR) for recurrence using the Cox proportional hazard
model. As shown in FIG. 9, taking the risk ratio for recurrence
of hepatic cancer to be 1 for the control group, this risk ratio
is approximately one thirdof that at 0. 329 for theVK-II-administered
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group; in particular, considering only the HCV cases, the risk ratio
becomes 0.210, i.e. is reduced to approximately one fifth, by
administering VK-TI.
FIG. 7 shows a graph indicating the results in the case of
excluding cases of local recurrence in the trial to verify the effect
of VK-II administration on hepatic cancer recurrence inhibition
(50o recurrence) (VK-II-administered group: 29 cases,
non-administered group: 22 cases) . Moreover, FIG. 8 shows a graph
indicating the results in the case of excluding cases of recurrence
within 6 months in the trial to verify the effect of VK-II
administration on hepatic cancer recurrence inhibition (500
recurrence) (VK-II-administered group: 31 cases, non-administered
group: 22 cases) . As shown in FIGS. 7 and 8, in these cases, again
the hepatic cancer cumulative recurrence rate was statistically
significantlysuppressedfor the VK-II-administered group compared
with the control group.
FIG. 10 shows graphs indicating the results of analyzing the
DCP level before treatment and upon recurrence. As shown in FIG.
10, with all of the cases of recurrence in the VK-II-administered
group, DCP was negative, and moreover there were no side effects,
and there were no dropout.
The action of VK-II according to the present application with
regard to invasion/metastasis of hepatic cancer cells was studied
in vitro. The action on invasion ability was investigated through
an invasion assay using HepG2 cells and a Matrigel chamber. The
results were that it was found that the number of cells passing
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through the Matrigel dropped upon adding VK-II, with the drop
depending on the concentration of the VK-II. Regarding the action
on the metastasis ability, the action of VK-II on the expression
of extracellular matrix metalloproteinases (MMPs) was investigated
using the Western blot method. Upon investigating the MMP-1 and
MMP-3 protein expression in the case of adding VK-II to hepatic
cancer cells, it was found that the expression was inhibited. From
the above, although the data is in vitro, it can be presumed that
VK-II inhibits invasion/metastasis of hepatic cancer cells.
Industrial Applicability
As described above, the menatetrenone-containing agent for
treating or preventing hepatic disease according to the present
invention has an excellent effect of inhibiting occurrence of PVI
with hepatic disease, in particular DCP positive hepatic cancer,
and moreover has an excellent effect of improving the prognosis
after hepatic cancer treatment.
Furthermore, the menatetrenone-containing agentfor treating
hepatic disease according to the present invention is very useful
in inhibiting recurrence of hepatic cancer after treatment.
1G
