U.S. patent application number 16/758022 was filed with the patent office on 2020-08-13 for therapeutic agent of uremia containing alarin as the main ingredient.
This patent application is currently assigned to JU TIDE CO., LTD.. The applicant listed for this patent is JU TIDE CO., LTD.. Invention is credited to Jong-Chul KIM.
Application Number | 20200254059 16/758022 |
Document ID | 20200254059 / US20200254059 |
Family ID | 1000004811536 |
Filed Date | 2020-08-13 |
Patent Application | download [pdf] |
![](/patent/app/20200254059/US20200254059A1-20200813-D00000.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00001.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00002.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00003.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00004.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00005.png)
![](/patent/app/20200254059/US20200254059A1-20200813-D00006.png)
![](/patent/app/20200254059/US20200254059A1-20200813-P00001.png)
![](/patent/app/20200254059/US20200254059A1-20200813-P00002.png)
United States Patent
Application |
20200254059 |
Kind Code |
A1 |
KIM; Jong-Chul |
August 13, 2020 |
THERAPEUTIC AGENT OF UREMIA CONTAINING ALARIN AS THE MAIN
INGREDIENT
Abstract
The present invention relates to a novel use of alarin derived
from a splice variant of galanin-like peptide (GALP) RNA and thus
having an amino acid sequence similar to that of galanin, with a
therapeutic agent of uremia, and the therapeutic agents of uremia
according to the present invention include alarin as the main
ingredient.
Inventors: |
KIM; Jong-Chul; (Yongin-si,
KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
JU TIDE CO., LTD. |
Seoul |
|
KR |
|
|
Assignee: |
JU TIDE CO., LTD.
Seoul
KR
|
Family ID: |
1000004811536 |
Appl. No.: |
16/758022 |
Filed: |
March 27, 2018 |
PCT Filed: |
March 27, 2018 |
PCT NO: |
PCT/KR2018/003601 |
371 Date: |
April 21, 2020 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/0019 20130101;
A61K 38/1709 20130101 |
International
Class: |
A61K 38/17 20060101
A61K038/17 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 26, 2017 |
KR |
10-2017-0179309 |
Claims
1. The therapeutic agent of uremia characterized in that the
therapeutic agent comprises alarin as a main component.
2. The therapeutic agent of uremia of claim 1, wherein the said
alarin is characterized in that it is the human-derived alarin
trifluoroacetate salt having the following amino acid sequence:
H-Ala-Pro-Ala-His-Arg-Ser-Ser-Thr-Phe-Pro-Lys-Trp-Val-Thr-Lys-Thr-Glu-Arg-
-Gly-Arg-Gln-Pro-Leu-Arg-Ser-OH-trifluoroacetate salt.
3. The therapeutic agent of uremia of claim 1, wherein the said
alarin has a molecular weight of 2894.29, characterized in that it
has the following molecular formula:
C.sub.127H.sub.206N.sub.43O.sub.38
4. The therapeutic agent of uremia of claim 1, wherein the said
therapeutic agent of uremia is characterized in that it is
formulated to be administered from 0.5 mg to 10 mg of alarin per an
adult of 60 kg body weight.
5. The therapeutic agent of uremia of claim 4, wherein the said
formulation is characterized in that it is an injectable one.
6. The therapeutic agent of uremia of claim 4, wherein the said
formulation is characterized in that it is injected by subcutaneous
muscle injection.
Description
TECHNICAL FIELD
[0001] The present invention relates to a therapeutic agent of
uremia containing alarin as the main ingredient, and more
particularly, to the use of alarin as a therapeutic agent of uremia
with a new use of alarin derived from a splice variant of
galanin-like peptide (GALP) RNA and having an amino acid sequence
similar to galanin.
BACKGROUND ART
[0002] Despite of the development of medical technology, there are
no effective method of treatments for many diseases yet, and such
intractable diseases may include uremia. Uremia refers to a
syndrome that shows various clinical symptoms resulting from
abnormal expression for functions of various organs and tissues
caused by harmful components of urine accumulated in blood, which
are occurred when the urine cannot be discharged outside the body
due to the breakdown of kidney function whatever the underlying
disease. The clinical symptoms of uremia include, for example,
digestive system abnormalities such as anorexia, nausea, vomiting,
bad breath, stomatitis, enteritis, nervous system abnormalities
such as indifference, indifference, decreased alertness, depressive
state, nausea, lethargy, circulatory system abnomial such as
anemia, red blood cell hematopoietic disorder, hypertension,
ischemic heart disease, pericarditis, myocarditis and the like, and
skin problems such as pigmentation, pruritus, subcutaneous
bleeding, and uremia is caused by the urea toxin substance
accumulated in the blood. There are various urea toxin substances,
for example, methyl guanidine, indole compounds, malondialdehyde,
creatinine, .beta.-aminoisolactic add, transketolase inhibitors,
polyamines, urea, phenols, and the like. However, it is still
unclear which of these urea substances are intrinsically involved
in the development of uremic symptoms, and it is unknown whether
uresis occurs only by a single substance or by a complex action of
these plural substances. However, it is reported that the latter is
most likely. These facts show clearly the difficulty of treating
uremia with medicine. Whatever its development of occurrence, a
person with uremia can face a fatal situation that leads to death,
unless the urinary substance is released out of the body or the
concentration of the urea in the blood is reduced by appropriate
means. Up to date, therapies that are effective in the treatment of
uremia include renal transplantation, dialysis and activated
charcoal agent therapies, and the benefits of the latter two common
treatments is that a plurality of urea toxin substances, including
unknown materials by physical means can be removed or diluted
nonspecifically.
[0003] However, activated charcoal agents are used as internal
medicines, and the mechanism of action of activated charcoal agent
is by adsorption and in vitro excretion of various toxic substances
generated in the digestive organs. Thus, activated charcoal agent
therapy is referred to as an internal medical agent, but like
dialysis therapy it can also be regarded as a physical therapeutic
means. In addition to that the activated charcoal agent is limited
for therapeutic effect, there are several problems such as
difficulty in taking them and a tendency to cause constipation, and
thus it are not very satisfactory treatments.
[0004] On the other hand, dialysis is the most effective treatment
for uremia, and in fact, when discharging the urea toxin substance
in vitro by dialysis therapy, there is an advantage that the death
of the patient due to the urea substance can be avoided even if the
kidney function is abolished. However, complications caused by
dialysis are becoming a new problem. These include, for example,
the development of anemia, nephrotic dystrophy, kidney cancer,
fibrinous pericarditis, aluminum osteopathy, amyloidosis,
crystalline arthritis, multiple cysts, and the like as
complications due to organ dialysis. In addition, there is a
problem in the medical economy such as the low quality of the
patient's social life and the occurrence of high treatment costs
that should visit frequently in the dialysis facility.
[0005] There has also been reported a method called "in vivo
dialysis" in which non-proteinaceous nitrogen but not any physical
method is transferred into the intestinal tract. It is the
phenomenon that, when the 15-keto-16-halogen-prostaglandin E2 class
is administered to the renal failure model rats, the blood serum
creatinine (Cre) and blood urea nitrogen (BUN) are transferred to
the organ together with the moisture by its enteropulling action
(action which store a moisture in the digestive tract). However,
the enteropulling action is an action of storing a moisture in the
digestive tract, and therefore, it inevitably involves diarrhea and
the reporter also mentions this point. Moreover, no matter how much
blood serum Cre or BUN values are lowered, long-term treatment of
this remedy which involves risk of physical depletion and
dehydration due to diarrhea is very burdensome for patients with
chronic renal failure and then is hard to be recognized as
practically acceptable treatment.
[0006] Therefore, there is a demand for the creation of a treatment
method of uremia or a therapeutic agent of uremia that does not
have the above-described problems. To meet these needs, under the
title of pharmaceutical composition useful for treating uremia,
Korean patent publication No. 1989-0001535 (Patent Document 1)
describes "the pharmaceutical composition containing a mixture of
several salts of the basic .alpha.-amino and .alpha.-keto analog of
the branched chain essential amino acid, the composition being
characterized in that it is provided in the form of a gastric
poorly solubility or gastric interoperability coating tablet".
[0007] However, the invention disclosed in the above Patent
Document 1 still has a problem in that it is difficult to provide a
satisfactory treatment effect for uremia, and thus there is still a
need for a therapeutic agent having excellent treatment effect of
uremia with long-term use without causing other side effects.
[0008] Meanwhile, alarin derived from a splice variant of
galanin-like peptide (GALP) RNA is present in vivo by biosynthesis
and synthesized with genetic engineering, which is a peptide having
the amino acid sequence of APAHRSSTFPKVWVTKTERGRQPLRS. Since these
alarin shares only 5 amino acids together with GALP, it cannot
activate GALP specific receptors, and neuropeptides have been
reported to be expressed in ganglion cells of human neuroblastoma
tumors and may be potential tumor markers. For more details on such
alarin, reference may be made to International Patent Publication
No. WO/2006/094973 (Patent Document 2), which are incorporated
herein by reference in their entirety.
[0009] The above Patent Document 2 only proposes the use of alarin
as a therapeutic agent for neurological keratitis, retinol disease,
acute and chronic inflammatory and autoimmune diseases, obesity or
growth deficiency, wound healing, uterine fibroids, endometriosis,
and anorexia.
[0010] Therefore, the present inventors have recognized and studied
to solve the above-mentioned conventional problems, and as a
result, we found that a peptide that can be simply provided, alarin
exerts an excellent improvement effect in cisplatin-induced uremia,
while being the peptide present in the human body do not cause side
effects of conventional therapeutic agents such as gastrointestinal
disorders, liver disorders, renal disorders, and diarrhea so that
can be used for a long time as a therapeutic agent for treating
uremia, so the present invention can be completed by the said
finding.
TECHNICAL PROBLEM
[0011] Accordingly, it is a main object of the present invention to
solve these problems of the prior art on the base of the background
of the problems in the conventional uremic treatments described at
the above. More specifically, an object of the present invention is
to provide a therapeutic agent of uremia that is able to long-term
use with providing a satisfactory treatment of uremia as a
therapeutic substance that can be simply provided, while not
causing side effects such as anemia, nephrotic dystrophy,
endocarditis, aluminum osteopathy, amyloidosis, crystalline
arthritis, multiple cysts, and diarrhea, and also incurring
qualitative deterioration and high treatment costs in the social
life of the patient.
[0012] Another object of the present invention is to provide a
method which can more easily provide a therapeutic agent of uremia
having the above-described characteristics.
[0013] The present invention may also be aimed at achieving, in
addition to the above-mentioned specific objects, other objects
that can be readily derived by those skilled in the art from the
said objects and the overall description of the present
specification.
TECHNICAL SOLUTION
[0014] In order to achieve the above object, the present inventors
have recognized the problems in the conventional treatment of
uremia and have studied to solve them. As a result, we obtained the
experimental data that the neuropeptide, alarin, which shares only
5 amino acids with GALP, has an excellent improvement effect in
cisplatin-induced uremia, and then reveal that it was possible to
provide a therapeutic agent of uremia, which is composed mainly of
alanine from the said results, and thus the object of the present
invention could be achieved.
[0015] The therapeutic agent of uremia according to the present
invention for achieving the above object is characterized in that
it comprises alarin as a main component.
[0016] According to another embodiment of the present invention,
the alarin is characterized in that it is the human-derived alarin
trifluoroacetate salt having the following amino acid sequence:
[0017]
H-Ala-Pro-Ala-His-Arg-Ser-Ser-Thr-Phe-Pro-Lys-Trp-Val-Thr-Lys-Thr-G-
lu-Arg-Gly-Arg-Gln-Pro-Leu-Arg-Ser-OH-trifluoroacetate salt.
[0018] According to another embodiment of the present invention,
the alarin has a molecular weight of 2894.29, characterized in that
it has the following molecular formula:
C.sub.127H.sub.206N.sub.43O.sub.38
[0019] According to another embodiment of the present invention,
the therapeutic agent of uremia according to the present invention
is characterized in that it is formulated to be administered from
0.5 mg to 10 mg of alarin per an adult of 60 kg body weight.
[0020] According to another embodiment of the invention, the
formulation is characterized in that it is an injectable one.
[0021] According to another embodiment of the invention, the
formulation is characterized in that it is injected by subcutaneous
muscle injection.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] The therapeutic agent of uremia containing alarin according
to the present invention will now be described by way of example
only, with reference to the accompanying drawings, in which:
[0023] FIG. 1 shows gene splice variants of galanin-like peptides
(GALPs) derived from Murine.
[0024] FIG. 2 shows the results of comparing the amino acid
sequences of alarin obtained from murine, rats, macaques and humans
according to a preferred embodiment of the present invention.
[0025] FIG. 3 is a graph showing the concentration of blood urea
nitrogen (BUN) in the control group and the group administered the
therapeutic agent of the present invention in Sprague-Dawley rat
model having a cisplatin-induced uremia according to example of the
present invention.
[0026] FIG. 4 is a graph showing the concentration of creatinine
(CRE) in the control group and the group administered the
therapeutic agent of the present invention in Sprague-Dawley rat
model having a cisplatin-induced uremia according to example of the
present invention.
[0027] FIG. 5 is a graph showing the histopathological assay
results in the control group and the group administered the
therapeutic agent of the present invention in Sprague-Dawley rat
model having a cisplatin-induced uremia according to example of the
present invention, wherein one point is less than 25% of coronary
necrosis at the junction site between the cortex and the cerebral
cortex, two points are from 25% to 50% of coronary necrosis, three
points are from 50% to 75% of coronary necrosis, and 4 points are
greater than 75% of coronary necrosis.
[0028] FIG. 6 is a graph showing the micrograph of
histopathological assay results in the control group and the group
administered the therapeutic agent of the present invention in
Sprague-Dawley rat model having a cisplatin-induced uremia
according to example of the present invention.
DETAILED DESCRIPTION OF THE INVENTION
[0029] Hereinafter, preferred embodiments of the present invention
will be described in detail.
[0030] The therapeutic agent of uremia according to a preferred
embodiment of the present invention contains alarin as a main
component.
[0031] According to another preferred embodiment of the present
invention, alarin used as a therapeutic agent of uremia of the
present invention may be human-derived alarin having the following
amino acid sequence:
[0032]
Ala-Pro-Ala-His-Arg-Ser-Ser-Thr-Phe-Pro-Lys-Trp-Val-Thr-Lys-Thr-Glu-
-Arg-Gly-Arg-Gln-Pro-Leu-Arg-Ser-OH
[0033] According to another preferred embodiment of the present
invention, alarin used as a therapeutic agent of uremia of the
present invention may be a pharmaceutically acceptable salt of
human-derived alarin having the said amino acid sequence.
[0034] By the said pharmaceutically acceptable salts are meant
salts which retain the biological effects of the free acids and
bases of specific compounds and are not biologically or otherwise
harmful. The compounds of the present invention may have functional
groups that are sufficiently acidic, sufficiently basic, or both,
and thus can react with any of the number of inorganic or organic
bases, and inorganic and organic acids to produce pharmaceutically
acceptable salts. Exemplary pharmaceutically acceptable salts
include salts prepared by reaction of a compound of the invention
with an inorganic or organic acid or inorganic base, for example
such as salts including trifluoroacetate, sulfate, pyrosulfate,
bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate,
dihydrogenphosphate, metaphosphate, pyrophosphate, chloride,
bromide, iodide, acetate, propionate, decanoate, caprylate,
acrylate, formate, isobutyrate, caproate, heptanoate, propiolate,
oxalate, malonate, succinate, cheverate, sebacate, fumarate,
maleate, butane-1,4-dioleate, hexain-1,6-dioleate, benzoate,
chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate,
methoxybenzoate, phthalate, sulfonate, xylenesulfonate,
phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate,
.gamma.-hydroxybutyrate, glycholate, tartrate, methane-sulfonate,
propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate
and mandelate.
[0035] According to another preferred embodiment of the present
invention, alarin used as a therapeutic agent of uremia of the
present invention may be a human-derived alarin trifluoroacetate
salt having the following amino acid sequence particularly:
[0036]
H-Ala-Pro-Ala-His-Arg-Ser-Ser-Thr-Phe-Pro-Lys-Trp-Val-Thr-Lys-Thr-G-
lu-Arg-Gly-Arg-Gln-Pro-Leu-Arg-Ser-OH-trifluoroacetate salt
[0037] According to another preferred embodiment of the invention,
the said alarin has a molecular weight of 2894.29 and has the
following molecular formula:
C.sub.127H.sub.206N.sub.43O.sub.38
[0038] According to another preferred embodiment of the present
invention, the said alarin may be stored stably at -20.+-.5.degree.
C.
[0039] According to another preferred embodiment of the present
invention, the therapeutic agent of uremia of the present invention
may be administered from 0.5 mg to 10 mg of alarin per adult of 60
kg body weight to provide a uremic treatment effect.
[0040] Preferably, the therapeutic agent of uremia of the present
invention may be administered from 1.0 mg to 5.0 mg of alarin per
adult of 60 kg body weight to provide a uremic treatment effect.
More preferably, the therapeutic agent of uremia of the present
invention may be administered from 2.0 mg to 5.0 mg of alarin per
adult of 60 kg body weight to provide a uremic treatment effect.
Even more preferably, the therapeutic agent of uremia of the
present invention may be administered from 3.0 mg to 5.0 mg of
alarin per adult of 60 kg body weight to provide a uremic treatment
effect. Most preferably, the therapeutic agent of uremia of the
present invention may be administered from 4.0 mg to 5.0 mg of
alarin per adult of 60 kg body weight to provide a uremic treatment
effect.
[0041] According to another preferred embodiment of the present
invention, the said therapeutic agent of uremia of the present
invention may be formulated into formulations which is intended to
be administered 0.5 mg to 10 mg, preferably 1.0 mg to 5.0 mg, more
preferably 2.0 mg to 5.0 mg, even more preferably 3.0 mg to 5.0 mg,
and most preferably 4.0 mg to 5.0 mg of alarin based on an adult of
60 kg body weight to provide a uremic treatment effect.
[0042] According to another preferred embodiment of the present
invention, the said formulation may be preferably, but not
particularly limited to, an injection.
[0043] According to another preferred embodiment of the present
invention, the said formulation may be preferably, but not
particularly limited to, an injectable one injected by subcutaneous
muscle injection.
[0044] According to another preferred embodiment of the present
invention, when the therapeutic formulation of the present
invention is an injection, a solvent, solubilizing agent or
emulsifying agent is used as the carrier component, such as water,
ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil,
glycerol aliphatic esters, polyethylene glycols or fatty acid
esters of sorbitan.
[0045] Hereinafter, the present invention will be described in more
detail with reference to preferred examples, but the present
invention is not limited to these examples.
EXAMPLE
[0046] Cisplatin-induced uremia was evaluated for the effect of
administration of test substance on an improvement of uremia in the
Sprague-Dawley rat model. Here, G1 is normal control group which is
administered with an excipient, G2 is induced control group which
is administered with an excipient, G3 is group which test substance
is administered with 10 .mu.g/1 mL/head, and G4 is group which test
substance is administered with 20 .mu.g/mL/head. Here, test
substance is alarin (human) trifluoroacetate salt of the present
invention. Cisplatin was intraperitoneally administered (IP) and
test substance was intravenously administered (IV).
[0047] In addition, the test substance (G3-G4) was administered
once at 8 hours (Day 0) after cisplatin administration, and in the
case of cisplatin (G2-G4), dose of 5 mg/10 mL/kg was administered
once at 8 hours (Day 0) before test substance administration.
[0048] The intravenous administration was done by fixing an animal
into a correction frame, and slowly injecting at a rate of within 1
mL/min through the micro vein using a syringe equipped with 26
gauges needle. The intraperitoneal administration was performed by
fixing the animal by a transdormal skin fixation method and
intraperitoneally injecting using a syringe equipped with 26 gauges
needle.
[0049] Body weights were measured on the day of administering test
substance, inducing and necropsy (Day 7). As a result, the weight
level of all the test substance-administered groups measured on the
day of necropsy showed no significant difference from the normal
control group.
[0050] In addition, blood is collected from the jugular vein of all
living animals prior to administration of the test substance,
before administration of cisplatin, and necropsy, injected into a
tube containing a coagulation activator, and allowed to coagulate
with standing at room temperature for about 15 minutes, followed by
centrifugation at 3,000 rpm for 10 minutes. Serum obtained from the
centrifugation was tested for blood urea nitrogen (BUN) and
creatine (CRE) using a blood biochemical analyzer (7180 Hitachi,
Japan). The blood biochemical test results showed significantly
lower BUN and CRE levels in the high dose group of the test
substance than the induced control group.
[0051] Finally, animals were inhaled with ether on an necropsy day
to anesthetize and euthanize, and kidneys were extracted and fixed
in a 10% neutral buffered formalin solution, and the fixed tissues
were cut, dehydrated, paraffin embedded and hematoxylin & eosin
stained to make a specimens for histopathological examination after
these histological treatment. And then, the obtained specimen is
subjected to speculum using an optical microscope (Olympus BX53,
Japan). Histopathological examination showed that the level of
lesion in the high dose group of test substance was significantly
lower than that of the control group.
[0052] Therefore, when the test substance is administered with a
single dose of 10 .mu.g/1 mL/head and 20 .mu.g/1 mL/head in a
Sprague-Dawley rat model in which uremia was induced by
administration of cystine under these test conditions, a tendency
to improve the blood biochemical level in the group of test
substance being compared to the control group of excipient was
observed, and in particular, the BUN and CRE levels of the test
substance of 20 .mu.g/1 mL/head showed a significant difference
compared to the control group of excipient. In addition, the
histopathological examination results showed a dose-related change
trend and significant changes in lesion level. Therefore, alarin of
the present invention is believed to have an effect on the
improvement of uremia in cisplatin-induced uremia model.
ADVANTAGEOUS EFFECTS
[0053] Since the therapeutic agent of uremia containing alarin as
the main ingredient of the present invention which is constituted
as described above is mainly composed of alarin which is a kind of
peptide that is present in vivo and can be simply synthesized, it
can be easily provided and shows an excellent uremic treatment
effect but also does not occur side effects such as anemia,
nephrotic dystrophy, epicarditis, aluminum osteopathy, amyloidosis,
crystalline arthritis, multiple cystic vesicles, diarrhea, and also
does not cause qualitative deterioration in the patient's social
life and high incidence of treatment costs accompanied with
treatment, thereby providing an excellent anti-uremic treatment
agent that can be used for a long time, and providing an excellent
effect to solve the problems associated with the conventional
treatment of uremia.
* * * * *