U.S. patent application number 09/969866 was filed with the patent office on 2002-04-04 for orally-administrable therapeutic and/or prophylactic agent for htlv-1-related diseases.
This patent application is currently assigned to KABUSHIKI KAISHA HAYASHIBARA SEIBUTSU KAGAKU KENKYUJO. Invention is credited to Kurimoto, Masashi, Ohashi, Kunihiro.
Application Number | 20020039570 09/969866 |
Document ID | / |
Family ID | 16570571 |
Filed Date | 2002-04-04 |
United States Patent
Application |
20020039570 |
Kind Code |
A1 |
Ohashi, Kunihiro ; et
al. |
April 4, 2002 |
Orally-administrable therapeutic and/or prophylactic agent for
HTLV-1-related diseases
Abstract
Disclosed are an orally-administrable therapeutic and/or
prophylactic agent for HTLV-1-related diseases, which comprises an
interferon-.gamma. as an effective ingredient and a
pharmaceutically-acceptable carrier, and a method for treating
and/or preventing the diseases with the agent. The HTLV-1-related
diseases include ATL, rheumatoid arthritis, Sjogren's syndrome,
SLE, uveitis, and immunopathies.
Inventors: |
Ohashi, Kunihiro; (Okayama,
JP) ; Kurimoto, Masashi; (Okayama, JP) |
Correspondence
Address: |
BROWDY AND NEIMARK, P.L.L.C.
SUITE 300
624 NINTH STREET, N.W.
WASHINGTON
DC
20001-5303
US
|
Assignee: |
KABUSHIKI KAISHA HAYASHIBARA
SEIBUTSU KAGAKU KENKYUJO
Okayamashi
JP
|
Family ID: |
16570571 |
Appl. No.: |
09/969866 |
Filed: |
October 4, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09969866 |
Oct 4, 2001 |
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09357913 |
Jul 21, 1999 |
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6299871 |
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Current U.S.
Class: |
424/85.5 ;
424/85.1; 424/85.2; 424/85.4; 435/343.1; 435/343.2; 435/5;
435/69.51 |
Current CPC
Class: |
A61K 38/217
20130101 |
Class at
Publication: |
424/85.5 ;
424/85.2; 424/85.1; 424/85.4; 435/69.51; 435/5; 435/343.1;
435/343.2 |
International
Class: |
A61K 038/21; C12Q
001/70; C12P 021/04; C12N 005/06; C12N 005/16; A61K 045/00; A61K
038/21 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 24, 1998 |
JP |
209294/1998 |
Claims
We claim:
1. An orally-administrable therapeutic and/or prophylactic agent
for HTLV-1-related diseases, which comprises an interferon-.gamma.
as an effective ingredient and a pharmaceutically-acceptable
carrier.
2. The agent of claim 1, wherein said interferon-.gamma. is of a
natural origin or one obtained by the recombinant DNA
technology.
3. The agent of claim 1, which further comprises, as a stabilizer
for said interferon-.gamma., one or more members selected from the
group consisting of saccharides, salts, amino acids, serum
albumins, gelatin, non-ionic surfactants, glucuronic acid,
dextrans, and hydroxyethyl starches.
4. The agent of claim 1, which contains about 0.1 to about 10.sup.6
units of said interferon-.gamma..
5. The agent of claim 1, which is in the form of a granule,
sugar-coated agent, troche or enteric-coated agent.
6. The agent of claim 1, wherein said HTLV-1-related diseases are
adult T-cell leukemia (ATL), Sjogren syndrome, chronic rheumatoid
arthritis, systemic lupus erythematosus (SLE), uveitis, and
immunopathies.
7. The agent of claim 1, which contains about 10 to about 10.sup.5
units of IFN-.gamma. per gram of the agent.
8. The agent of claim 1, which is in a dose unit form and which
contains an about 0.1 to about 10.sup.6 units of the
IFN-.gamma..
9. A method for treating and/or preventing HTLV-1-related diseases,
comprising a step of administering the agent of claim 1 to a
subject suffering from an HTLV-1-related disease.
10. The method of claim 9, wherein said HTLV-1-related disease is a
member selected from the group consisting of adult T-cell leukemia
(ATL), Sjogren syndrome, chronic rheumatoid arthritis, systemic
lupus erythematosus (SLE), uveitis, and immunopathies.
11. The method of claim 9, wherein said agent is administered to
said subject at a dose of one to 1.times.10.sup.4
units/adult/day/kg of the body weight.
12. The method of claim 9, wherein said agent is administered to
said subject at a dose frequency of 1-4 shots/day or 1-7 shots/week
over one week to one year.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to an orally-administrable
therapeutic and/or prophylactic agent for HTLV-1 (Human T-cell
Lymphotropic Virus Type 1) related diseases, and more particularly
to an orally-administrable therapeutic and/or prophylactic agent
for HTLV-1-related diseases, which comprises an interferon-.gamma.
(abbreviated as IFN-.gamma. hereinafter) as an effective
ingredient.
[0003] 2. Description of the Prior Art
[0004] Adult T-cell leukemia (abbreviated as ATL hereinafter) is a
T-cell leukemia with characteristic symptoms, found and reported by
Cache TAKATSUKI in 1976. The disease is an intractable and
district-specific disease, and in some cases it may cause a
malignant lymphoma-like symptom; In Japan, there found many
patients in the South and East Japan including Kyushu-, Okinawa-
and Shikoku-Islands. In overseas, in the tropical regions such as
the Caribbean Sea and the South India. Most of the leukemia cells
induced by ATL have CD4-positive and CD8-negative helper T-cell
surface antigens, and show a specific change in nuclear. It was
revealed that ATL is induced by HTLV-1, a C-type retrovirus, as a
causative virus thereof.
[0005] Epidemiological research revealed that HTLV-1-infected
patients or HTLV-1 carriers may cause not only ATL but induce
malignant tumors at a relatively-high efficiency, as well as
neuropathies and immunopathies called HAM (HTLV-1-associated
myelopathy). HTLV-1 is now being researched that, in addition to
ATL, it correlates to another diseases. Furthermore, it is pointed
out that HTLV-1 may relate to chronic rheumatoid arthritis, Sjogren
syndrome, systemic lupus erythematosus (SLE), uveitis, etc.
[0006] HTLV-1 infection can be easily diagnosed by detecting an
HTLV-specific antibody in blood and by confirming the presence of
an HTLV-1 proviral DNA. Symptoms and developments of
HTLV-1-infected patients are varied, and the patients may become to
show the symptoms at their age of 50th. Physiologically, there
observed the downfall of myelin sheathes and the loss of axons from
the upper part of cervicals through lumbars, the invasion of
mononuclear cells such as lymphocytes and macrophages, the
proliferation of astrogrias, and a slight level of the mononuclear
cells' invasion in the part of brain stem and the substantia alba
of cerebrum and cerebellum.
[0007] The percentage of the incidence of ATL induced by HTLV-1 is
relatively low. However, once occurred, the symptom of the patients
infected with the virus is rapidly worsened, and the treatment is
quite difficult. Conventional therapies include the administration
of a relatively-large amount of adrenal cortical hormone, and
chemical- and radio-therapies in accordance with those for
malignant tumors. They are, however, merely temporal symptomatic
therapies which are far from intrinsic therapy. As another
clinical-test-treatment for ATL as an HTLV-1-related disease,
Kazuyuki ISHIHARA proposed in Skin Cancer, Vol. 12, No. 2, pp.
301-314 (1997) an intramuscular injection of IFN-.gamma., where
several millions units of IFN-.gamma. are injected to a patient
daily for over eight weeks. In such a treatment, patients are
forced to stay in hospitals or allowed to go to hospitals
frequently, resulting in physical- and mental-pains and economical
burdens. In the latter case, when patients unexpectedly could not
go to hospitals on their prescribed administration dates, the
medicinal administration control is not sufficiently conducted, and
as a drawback, this hinders the expected therapeutic effect.
Although IFN-.gamma. is per se a safer medicine which scarcely
induces side effects even when administered to patients at a
relatively-high dose, the above daily high-dose as much as several
hundreds units of IFN-.gamma. may result in side effects such as
serious depression of liver function, leukopenia, neutropenia,
calcium lowering, and fervescence. In the most serious case, the
administration must be ceased.
[0008] Under these backgrounds, greatly expected is the
establishment of a relatively-safe medicine which can intrinsically
and effectively treat and/or prevent HTLV-1-related diseases, and
lower the patients' mental, physical, and economical burdens.
SUMMARY OF THE INVENTION
[0009] The present invention aims to provide a medicament with
lesser side effects, which can intrinsically and effectively treat
and/or prevent HTLV-1-related diseases.
[0010] The present inventors energetically researched on
medicaments which can intrinsically and effectively treat and/or
prevent HTLV-1-related diseases, and on the administration route.
As a result, they solved the above object by establishing an
orally-administrable therapeutic and/or prophylactic agent for
HTLV-1-related diseases, which comprises an IFN-.gamma. as an
effective ingredient. As a characteristic feature of the present
invention, the oral administration of IFN-.gamma. as an effective
ingredient more effectively treats and/or prevents HTLV-1-related
diseases at an extremely-lower dose than another
administrations.
DETAILED DESCRIPTION OF THE INVENTION
[0011] Explaining the preferred embodiments of the present
invention, the IFN-.gamma., which is incorporated as an effective
ingredient into the present orally-administrable therapeutic and/or
prophylactic agent for HTLV-1-related diseases, includes natural
IFN-.gamma.s produced from IFN-.gamma.-producing human leukocytes
and established cell lines, and recombinant IFN-.gamma.s obtained
by introducing genes for encoding the above IFN-.gamma.s into
animal cells or microorganisms such as the species Escherichia Coli
to transform them by the recombinant DNA technology. These
IFN-.gamma.s arbitrarily used in the present invention include not
only those in a highly-purified form having a specific activity of
at least 1.times.10.sup.7 units/mg protein but those in a crude
form which contain merely pharmaceutically-acceptable impurities as
long as they attain the present object. Because the present agent
is orally administrable, it does not necessarily require the
highest possible purity of IFN-.gamma. requisite for intramuscular-
and intravenous-injections, and it can be arbitrarily prepared even
with a relatively-low purity IFN-.gamma.. Using these IFN-.gamma.s,
the present orally-administrable agent can be prepared at a
relatively-low cost. In the present invention, two or more types of
IFN-.gamma.s can be used as the IFN-.gamma.s. With a viewpoint of
antigenicity, human IFN-.gamma.s, in particular, natural types of
IFN-.gamma.s, can be more advantageously used.
[0012] Into the present orally-administrable agent can be
arbitrarily incorporated one or more pharmaceutically-acceptable
diluents, excipients, fillers, stabilizers, pH-controlling agents,
biologically-active substances, etc., in addition to the
IFN-.gamma. as an effective ingredient.
[0013] The stabilizers used in the present invention mean agents
capable of stabilizing the IFN-.gamma.; saccharides and sugar
alcohols such as glucose, galactose, xylose, fructose, sucrose,
maltose, trehalose, neotrehalose, isotrehalose, sorbitol, mannitol,
maltitol, lactitol, lactosucrose, maltooligo-saccharides, and
polysaccharides; cyclodextrins, hydroxyethyl starches, dextrins,
and dextrans; salts such as sodium glucronate, phosphates, and
metal salts; and serum albumin, gelatin, amino acids, and non-ionic
surfactants. Among these stabilizers, maltose and trehalose stably
retain the present-effective ingredient(s) for a relatively-long
period of time.
[0014] Except for the non-ionic surfactants, the percentage of the
above stabilizers which are incorporated into the present
orally-administrable agent is not specifically restricted;
Generally, it is in the range of about 0.01 to below 100 w/w %,
preferably, about 0.1 to below 100 w/w %, and more preferably about
1 to below 100 w/w %, based on the weight of the present
orally-administrable agent. The percentage of the non-ionic
surfactants is from one microgram to one milligram, and preferably
10 micrograms to one milligram per gram of the present
orally-administrable agent. Depending on the form of the present
orally-administrable agent, the addition of the stabilizers in an
amount within the above range stably retains the activities of the
IFN-.gamma. for, in general, at least 36 months at 4.degree. C.
conditions, and for at least six months even at ambient
temperature. In the case of using maltose and/or trehalose as the
stabilizers, the IFN-.gamma. can be usually retained stably for at
least 12 months even at ambient temperature. While the other
stabilizers tend to have an inferior stabilizing-activity to that
attained by maltose and/or trehalose.
[0015] The content of the IFN-.gamma., which is incorporated as an
effective ingredient into the present orally-administrable agent,
is an amount sufficient for effectively reducing the HTLV-1 virus
level and its antibody level. Usually, it is from about 0.1 to
about 10.sup.9 units; preferably, from about one to about 10.sup.7
units; more preferably, from about 10 to about 10.sup.5 units; and
most preferably from about 10.sup.2 to about 10.sup.4 units per
gram of the present orally-administrable agent. When the present
agent is in a dose unit form, it should preferably contain an about
0.1 to about 10.sup.6 units, preferably, about 10 to about 10.sup.5
units of the IFN-.gamma..
[0016] The form of the present orally-administrable agent is
selected from those in orally-administrable forms of liquids,
pastes, troches, capsules, powders, granules, tablets, and enteric
coated agents. In the case of formulating the present
orally-administrable agent into a liquid or paste form, the agent
thus obtained is adjusted to a pH that does not inactivate the
IFN-.gamma. as the present effective ingredient, i.e., pHs of from
about 4 to about 9, and preferably pHs of 6-8. The present
orally-administrable agent should preferably be stored at 4.degree.
C. under dark conditions to stably keep the activity and quality,
independently of its form and shape. Since the IFN-.gamma. is
unstable under strong-acid conditions, the present
orally-administrable agent should preferably be formulated into a
granule, sugar-coated agent, capsule or enteric-coated agent to
prevent the effective ingredients from being inactivated by gastric
acid after orally administered. Particularly, the present
orally-administrable agent in the form of an enteric-coated agent
is most preferable. The processing method for formulating the
present orally-administrable agent into various shapes and forms
should not be restricted, and it can be appropriately selected from
conventional ones.
[0017] Explaining the dose of the present orally-administrable
therapeutic and/or prophylactic agent for HTLV-1-related diseases,
in case of the present agent containing the IFN-.gamma. as an
effective ingredient, the oral dose of IFN-.gamma. is one to
1.times.10.sup.4 units/adult/day/kg of the body weight; preferably,
1.times.10 to 1.times.10.sup.4 units/adult/day/kg of the body
weight; and more preferably, 1.times.10.sup.2 to 1.times.10.sup.3
units/adult/day/kg of the body weight, at 1-4 shots/day or 1-7
shots/week over one week to one year. In the case of the present
orally-administrable agent, which comprises the IFN-.gamma. is in a
unit dose form, the effective ingredients are administered to
subjects within the above dose range. The administration period is
not specifically restricted and can be arbitrarily set depending on
the types, contents and doses of the effective ingredients, and
patients' symptoms; usually, 1-24 months, preferably, 3-18 months,
and more preferably, 6-12 months.
[0018] The following Experiments and Examples describe the present
invention in more detail:
[0019] Experiment 1
[0020] Treatment of Adult T-cell Leukemia
[0021] Eight HTLV-1 carriers, who were infected with HTLV-1 but not
yet showed the symptoms, were divided into Groups A, B and C
consisting of three, three and two subjects, respectively. The
subjects of Group A were administered with the present
orally-administrable agent, 200 mg per tablet, containing 1,000
units of IFN-.gamma., obtained by the later described method in
Example 1, while the subjects of Group B were administered with
another type of the present orally-administrable agent, 200 mg per
tablet, containing 10,000 units of IFN-.gamma., obtained by the
later described method in Example 2, at a dose of 3
shots/day/subject over 12 months. The subjects of Group C were
orally administered similarly as above with an orally-administrable
agent, 200 mg per tablet, free of IFN-.gamma. and consisting of a
base. Every patient was sampled their blood before the
administration tests and sampled at 3rd, 6th, 9th and 12th months
after initiation of the tests to determine the virus level of
HTLV-1 and the level of antibody against HTLV-1. The levels of the
virus and antibody were respectively determined by conventional
hybridization method and EIA method (enzyme immunosolvent assay).
The virus level was expressed with a relative value (%) based on
the level before the administration test being regarded as 100%.
The antibody level was expressed with the symbols "++", "+" and "-"
which meant "high", "low", and "below the detectable level",
respectively. The results are in Table 1.
1 TABLE 1 Before After administration Group administration 3rd
month 6th month 9th month 12th month A No. 1 Virus level 100% 100%
80% 30% 10% (Present invention) Antibody level + + + + + + + + + +
No. 2 Virus level 100% 80% 1% 0.01% 0% Antibody level + + + + + + +
No. 3 Virus level 100% 60% 0.5% 0.1% 0.01% Antibody level + + + + +
+ + + + B No. 1 Virus level 100% 15% 0.01% 0% 0% (Present
invention) Antibody level + + + + + + + - No. 2 Virus level 100%
20% 0.01% 0% 0% Antibody level + + + + + + + - No. 3 Virus level
100% 50% 3% 0.01% 0% Antibody level + + + + + + + - C No. 1 Virus
level 100% 100% 100% 102% 105% (Control) Antibody level + + + + + +
+ + + + No. 2 Virus level 100% 100% 100% 101% 102% Antibody level +
+ + + + + + + + +
[0022] In Groups A and B administered with the present
orally-administrable agent of IFN-.gamma., the HTLV-1 virus level
was lowered depending on the IFN-.gamma. concentration. In
particular, after the administration tests, no HTLV-1 was detected
in the subjects of Group A, No. 2, and of Group B, Nos. 1 and 2,
and no HTLV-1 was detected in them even six months after the tests.
The data strongly indicates that HTLV-1 was completely eliminated
from these subjects. In Groups A and B, there found some subjects
who lowered leukocytes in number which might be caused by
IFN-.gamma., but after the oral-administration test all of them
recovered to their normal levels within a relatively-short period
of time. In the groups with IFN-.gamma., no subject complained
about side effects such as fervescence, languor, and lack of
appetite.
[0023] These results indicate the effectiveness and safety of the
present orally-administrable therapeutic and/or prophylactic agent
for HTLV-1-related diseases.
[0024] Experiment 2
[0025] Treatment of Adult T-cell Leukemia
[0026] Three patients, who were suffering from adult T-cell
leukemia and found out with skin erythema characteristic of ATL,
were orally administered with an orally-administrable agent, 200 mg
per tablet, according to the present invention, which contained
1,000 units of the later described IFN-.gamma. obtained in Example
1, at a dose of thrice-a-day for six months. As a control, two
patients, who were observed with skin erythema characteristic of
ATL, were orally administered with an orally-administrable agent,
200 mg per tablet, free of IFN-.gamma. and consisting of a base,
prepared according to the method in Example 1. Every patient was
macroscopically observed his or her skin erythema before initiating
the administration, and at 2nd and 4th months and 6th month
(completion of the administration) after the administration, and
examined for HTLV-1 virus level in blood, level of antibody against
HTLV-1, leukocyte number, and percentage (%) of abnormal
lymphocytes to normal ones. The patients were asked for examining
side effects such as fervescence, languor, and lack of appetite.
The assay for virus and antibody levels, and the evaluation of the
antibody level were conducted similarly as in Experiment 1. The
evaluation of the degree of skin erythema was expressed with the
symbols "++", "+" and "-" which meant that the levels or symptoms
of skin erythema were "high", "low", and "diminished",
respectively. The results are in Table 2.
2 TABLE 2 Before After administration Test No. administration 2nd
month 4th month 6th month No. 1 Virus level 100% 100% 100% 100%
(Present Antibody level ++ ++ ++ ++ invention) Skin erythema level
++ ++ + - Hemateikon leukocyte 11000 11000 10000 10000
(cells/mm.sup.3) Abnormal 5.3% 5.2% 4.5% 2.3% lymphocyte (%) Side
effects Fervescence Non Non Non Non Languor Non Non Non Non Lack of
appetite Non Non Non Non No. 2 Virus level 100% 60% 0.5% 0.1%
(Present Antibody level ++ ++ ++ ++ invention) Skin erythema level
++ ++ ++ + Hemateikon leukocyte 12500 12000 12000 11500
(cells/mm.sup.3) Abnormal 5.8% 5.5% 5.4% 5.2% lymphocyte (%) Side
effects Fervescence Non Non Non Non Languor Non Non Non Non Lack of
appetite Non Non Non Non No. 3 Virus level 100% 80% 1% 0.01%
(Present Antibody level ++ + - - invention) Skin erythema level ++
+ + - Hemateikon leukocyte 12000 12000 12000 11500 (cells/mm.sup.3)
Abnormal 6.8% 6.5% 6.4% 6.2% lymphocyte (%) Side effects
Fervescence Non Non Non Non Languor Non Non Non Non Lack of
appetite Non Non Non Non No. 4 Virus level 100% 101% 102% 102%
(Control) Antibody level ++ ++ ++ ++ Skin erythema level ++ + + -
Hemateikon leukocyte 11500 11500 11200 11000 (cells/mm.sup.3)
Abnormal 5.5% 5.9% 5.9% 6.0% lymphocyte (%) Side effects
Fervescence Non Non Non Non Languor Non Non Non Non Lack of
appetite Non Non Non Non No. 5 Virus level 100% 102% 103% 104%
(Control) Antibody level ++ ++ ++ ++ Skin erythema level ++ ++ ++
++ Hemateikon leukocyte 12000 12200 12100 12000 (cells/mm.sup.3)
Abnormal 5.0% 5.2% 5.4% 5.6% lymphocyte (%) Side effects
Fervescence Non Non Non Non Languor Non Non Non Non Lack of
appetite Non Non Non Non
[0027] As shown in Table 2, depending on the administration period
of IFN-.gamma., the patients in the groups with the present
orally-administrable agent were lowered or reduced in the level of
HTLV-1 virus, antibody, skin erythema, and abnormal lymphocyte.
These patients were slightly lowered in leukocyte number which
might be induced by IFN-.gamma., but after completion of the
IFN-.gamma. administration they all recovered their normal
leukocyte levels within a relatively-short period of time. In the
group with the orally-administrable agent of IFN-.gamma., no
patient complained about side effects such as fervescence, languor,
and lack of appetite.
[0028] These results show the effectiveness and safety of the
present orally-administrable therapeutic and/or prophylactic agent
for HTLV-1-related diseases.
[0029] Experiment 3
[0030] Treatment of Sjoqren's Syndrome
[0031] Eight patients with Sjogren's syndrome were divided into two
groups which consisted of four subjects each. Patients in one group
were orally administered with the present orally-administrable
agent, 200 mg per tablet, containing 1,000 units of IFN-.gamma.
obtained by the later described method in Example 1, at a dose of
thrice-a-day over six months, while those in another group were
orally administered with an orally-administrable agent, 200 mg per
tablet, free of IFN-.gamma. and consisting of a base, prepared
similarly as above. With an index of the lowering of salivary
secretion characteristic of patients with the syndrome, the
secretion volume before and after the administration was measured
by the Saxon method to evaluate the affectivity of the present
agent. The results are in Table 3.
3 TABLE 3 Salivary secretion (g/min) Present invention Control
(Group with IFN-.gamma.) (Group with no IFN-.gamma.) Before 0.62
.+-. 0.43 0.56 .+-. 0.34 administration After 1.23 .+-. 0.65 0.45
.+-. 0.38 administration
[0032] As evident from the results in Table 3, the patients of the
group with the present IFN-.gamma. orally-administrable agent
significantly increased in salivary secretion. In the control, no
increment of salivary secretion was found.
[0033] The data indicates that the present orally-administrable
therapeutic and/or prophylactic agent for HTLV-1-related diseases
is strongly effective on Sjogren's syndrome.
[0034] Experiment 4
[0035] Immuno-activatinq Activity
[0036] Four healthy volunteers, consisting of two males and females
each, were divided into two groups consisting of both sexes. The
subjects, Nos. 1 and 2, in one group were orally administered with,
as the present agent, an orally-administrable agent, 200 mg per
tablet, containing 100 units of an IFN-.gamma. prepared in
accordance with the later described method in Example 1, at a dose
of thrice-a-day over six months. The subjects, Nos. 3 and 4, in
another group were orally administered with an orally-administrable
agent, 200 mg per tablet, free of IFN-.gamma. and consisting of a
base, which had been prepared similarly as above agent. The
patients in each group were sampled their blood before and
two-weeks after the administration for assaying NK-cell activity
and tumor-cell-killing activity of mononuclear cells.
[0037] The NK cell activity was assayed by separating lymphocytes
from a sampled blood, and evaluating the activity based on the
cytotoxic effect by the lymphocytes against K-562 cell, ATCC CCL
243, human chronic myelogenous leukemia; Peripheral lymphocytes as
effector cells and .sup.51Cr-labeled K-562 cells as target cells
were mixed at different ratios of effector cell/target cell (E/T
ratio) and cultured for four hours, followed by measuring the
radio-activity of .sup.51Cr released in the supernatant. The
results are in Table 4. The increment of the radio-activity means
that the present orally-administrable agent activated NK cells as
immunocompetent cells.
4 TABLE 4 Present invention Control (Group with (Group with
IFN-.gamma.) no IFN-.gamma.) Volunteer No. No. 1 No. 2 No. 3 No. 4
Radio- Before 18.4 26.5 16.3 20.3 activity administration After
24.4 30.4 17.4 19.7 administration
[0038] As evident from the results in Table 4, in the control,
there found no increment of NK cell activity but found a remarkable
increment in the group with the present orally-administrable
therapeutic and/or prophylactic agent for HTLV-1-related
diseases.
[0039] The cell-killing activity by mononuclear cells was measured
by separating mononuclear cells from the volunteers' blood, and
evaluating the activity based on the cytotoxic reaction against
A375.S2, ATCC CRL-1872, human malignant melanoma; It was measured
by mixing the mononuclear cells as effector cells and
[.sup.125]uridine-labeled A375.S2 cells as target cells in a ratio
of effector cells to target cells (E/T ratio) of 20, incubating the
cells for 72 hours, and measuring the radio-activity of the
remaining target cells for determining the cell-killing activity of
the mononuclear cells. The results are in Table 5. The increment of
radio-activity means that the present orally-administrable agent
activated the cell killing activity by mononuclear cells as
immunocompetent cells.
5 TABLE 5 Present invention Control (Group with (Group with
IFN-.gamma.) no IFN-.gamma.) Volunteer No. No. 1 No. 2 No. 3 No. 4
Radio Before 1.2 2.4 2.3 1.3 activity administration After 3.4 5.3
2.1 1.2 administration
[0040] As evident from the results in Table 5, in the control,
there found no increment of cell killing activity by human
mononuclear cells but found a remarkable increment in the group
with the present orally-administrable therapeutic and/or
prophylactic agent for HTLV-1-related diseases.
[0041] The following Examples describe the preferred embodiments
according to the present invention in more detail:
EXAMPLE 1
[0042] Orally-administrable Agent
[0043] A natural human IFN-.gamma. having a specific activity of
about 1.times.10.sup.7 units/mg protein commercialized by
Hayashibara Biochemical Laboratories, Inc., Okayama, Japan, and
"TREHAOSE.RTM.", a trehalose commercialized by Hayashibara Shoji,
Inc., Okayama, Japan, were mixed, and the mixture was tabletted in
a usual manner into a crude tablet, 180 mg, containing
1.times.10.sup.3 units of IFN-.gamma.. A base for enteric coating,
consisting of 100 g hydroxypropylmethylcellulose phthalate (HPMCP),
500 ml ethanol, and 100 of refined water, was prepared. The above
crude tablet was coated with the base into an enteric coated
tablet, 200 mg, as the present orally-administrable agent.
[0044] Since the product has, as an effective ingredient, an
IFN-.gamma. stabilized by trehalose and is enteric coated, the
decomposition of IFN-.gamma. by gastric acid is well prevented. The
product can be arbitrarily used in the treatment and/or the
prevention of ATL, rheumatoid arthritis, Sjogren's syndrome, SLE,
uveitis, and immunopathies. Especially, ATL can be effectively
treated with the product.
EXAMPLE 2
[0045] Orally-administrable Agent
[0046] A natural human IFN-.gamma. having a specific activity of
about 1.times.10.sup.7 units/mg protein commercialized by
Hayashibara Biochemical Laboratories, Inc., Okayama, Japan;
"FINETOSE.RTM.", a crystalline maltose anhydride commercialized by
Hayashibara Shoji, Inc., Okayama, Japan; and "TREHAOSE.RTM.", a
trehalose commercialized by Hayashibara Shoji, Inc., Okayama,
Japan, were mixed, and the mixture was tabletted in a usual manner
into a crude tablet, 170 mg, containing 1.times.10.sup.4 units of
IFN-.gamma.. A base for enteric coating, consisting of 200 g acetic
acid cellulose phthalate (CPA), 500 ml ethanol, and 100 of refined
water, was prepared. The above crude tablet was coated with the
base into an enteric coated tablet, 200 mg, as the present
orally-administrable agent.
[0047] Since the product has, as an effective ingredient, an
IFN-.gamma. stabilized by maltose and trehalose and is enteric
coated, the decomposition of IFN-.gamma. by gastric juice is well
prevented. The product can be arbitrarily used in the treatment
and/or the prevention of ATL, rheumatoid arthritis, Sjogren's
syndrome, SLE, uveitis, and immunopathies. Especially, ATL can be
effectively treated with the product.
EXAMPLE 3
[0048] Orally-administrable Agent
[0049] In 1,000 ml (pH 7.0) of a physiological saline containing
both 100 .mu.g/ml human serum albumin and 10 w/v % maltose was
dissolved 5,000 units of a recombinant human IFN-.gamma. with a
specific activity of about 1.times.10.sup.6 units/mg protein,
commercialized by Japan Chemical Research, Tokyo, Japan. The
resulting solution was in a usual manner filtered with a membrane
filter to obtain an orally-administrable agent as the present
orally-administrable agent. The dose of the product is in the range
of 10-100 ml/shot.
[0050] Since the product is stabilized by maltose, i.e., the
IFN-.gamma. as an effective ingredient is stably retained over a
period of 36 months under 4.degree. C. conditions, it can be
arbitrarily used in the treatment and/or the prevention of ATL,
rheumatoid arthritis, Sjogren's syndrome, SLE, uveitis, and
immunopathies. Especially, ATL can be effectively treated with the
product.
EXAMPLE 5
[0051] Orally-administrable Agent
[0052] A physiological saline containing 5 W/V % "FINETOSE.RTM.", a
crystalline maltose anhydride commercialized by Hayashibara Shoji,
Inc., Okayama, Japan; 5 w/v % Y "TREHAOSE.RTM.", a trehalose
commercialized by Hayashibara Shoji, Inc., Okayama, Japan; and one
w/v % gelatin was prepared and allowed to dissolve a natural human
IFN-.gamma. with a specific activity of about 1.times.10.sup.7
units/mg protein to give a concentration of 10 units/ml of
IFN-.gamma.. The solution was in a usual manner filtered with a
membrane filter for sterilization to obtain an orally-administrable
agent as the present orally-administrable agent. The dose of the
product is in the range of 10-50 ml/shot. Since the product is
stabilized by maltose and trehalose where the IFN-.gamma. as an
effective ingredient is stably retained over a period of 36 months
under 4.degree. C. conditions, it can be arbitrarily used in the
treatment and/or the prevention of ATL, rheumatoid arthritis,
Sjogren's syndrome, SLE, uveitis, and immunopathies. Especially,
ATL can be effectively treated with the product.
[0053] As described above, the present invention was made based on
the finding that, when orally administered to patients with
HTLV-1-related diseases, IFN-.gamma. effectively treats and/or
prevents ATL, rheumatoid arthritis, Sjogren's syndrome, SLE,
uveitis, and immunopathies within a relatively-short period of
time. The present orally-administrable agent intrinsically treats
and/or prevents HTLV-1-related diseases which have been deemed
substantially impossible of the treatment and/or the prevention
with conventional adrenocorticotropic hormones, and chemical- and
radio-therapies. The present agent most effectively acts on
HTLV-1-related diseases, particularly, ATL, and can completely
eliminate the virus from the body of HTLV-1-carriers
[0054] The present invention with such outstanding effects will
greatly contribute to this art.
[0055] While there has been described what is at present considered
to be the preferred embodiments of the invention, it will be
understood the various modifications may be made therein, and it is
intended to cover the appended claims all such modifications as
fall within the true spirits and scope of the invention.
* * * * *