U.S. patent application number 15/236055 was filed with the patent office on 2017-02-16 for compound for treating sequelae of ischemic cerebral stroke.
The applicant listed for this patent is Jingyi Wang. Invention is credited to Xiaoliang Lv, Jingyi Wang, Tan Zhang, Jianling Zuo.
Application Number | 20170044212 15/236055 |
Document ID | / |
Family ID | 56686692 |
Filed Date | 2017-02-16 |
United States Patent
Application |
20170044212 |
Kind Code |
A1 |
Wang; Jingyi ; et
al. |
February 16, 2017 |
COMPOUND FOR TREATING SEQUELAE OF ISCHEMIC CEREBRAL STROKE
Abstract
The present invention provides a compound (I) for treating
sequelae of ischemic cerebral stroke:
H--(NH--CHR.sub.1--CO)--(NH--CHR.sub.2--CO)--(NH--CHR.sub.3--CO)--(NH--C-
HR.sub.4--CO)--(NH--CHR.sub.5--CO)--(NH--CHR.sub.6--CO)--(NH--CHR.sub.7--C-
O)--OH (I) or a pharmaceutically acceptable salt thereof, wherein
R.sub.1-R.sub.7 are defined herein. The present invention also
provides a pharmaceutical composition comprising said compound and
use of the same in the manufacture of a medicament for treating
sequelae of ischemic cerebral stroke. The compound and
pharmaceutical composition according to the present invention have
good pharmacological activities so that they are able to improve
significantly the symptom of sequelae of ischemic cerebral
stroke.
Inventors: |
Wang; Jingyi; (Suzhou,
CN) ; Zuo; Jianling; (Suzhou, CN) ; Lv;
Xiaoliang; (Suzhou, CN) ; Zhang; Tan; (Haikou,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Wang; Jingyi |
Suzhou |
|
CN |
|
|
Family ID: |
56686692 |
Appl. No.: |
15/236055 |
Filed: |
August 12, 2016 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
PCT/CN2015/087048 |
Aug 14, 2015 |
|
|
|
15236055 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 38/00 20130101;
A61K 38/08 20130101; A61P 9/10 20180101; C07K 7/06 20130101 |
International
Class: |
C07K 7/06 20060101
C07K007/06; A61K 38/08 20060101 A61K038/08 |
Claims
1. A compound of formula (I):
H--(NH--CHR.sub.1--CO)--(NH--CHR.sub.2--CO)--(NH--CHR.sub.3--CO)--(NH--CH-
R.sub.4--CO)--(NH--CHR.sub.5--CO)--(NH--CHR.sub.6--CO)--(NH--CHR.sub.2--CO-
)--OH (I) wherein each of R.sub.1, R.sub.4 and R.sub.6 is
independently selected from the group consisting of C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; R.sub.2
is C.sub.1-C.sub.6 alkyl which is substituted with five to ten
membered heteroaryl optionally substituted with halo, hydroxyl,
sulfydryl, carboxyl, amino, nitro, or cyano; R.sub.3 is
C.sub.1-C.sub.6 alkyl group substituted by halo, hydroxyl,
sulfydryl, or amino; R.sub.5 is C.sub.1-C.sub.6 alkyl substituted
with carbamoyl; and R.sub.7 is C.sub.1-C.sub.6 alkyl which is
substituted with C.sub.6-C.sub.14 aryl optionally substituted with
halo, hydroxyl, sulfydryl, carboxyl, amino, nitro, or cyano; or a
pharmaceutically acceptable salt thereof.
2. The compound or a pharmaceutically acceptable salt thereof
according to claim 1, wherein the heteroaryl is selected from five
to six membered heteroaryl.
3. The compound or the pharmaceutically acceptable salt thereof
according to claim 1, wherein the five to ten membered heteroaryl
is selected from the group consisting of pyrrolyl, pyrazolyl,
imidazolyl, oxazolyl, thienyl, iso-oxazolyl, oxadiazolyl,
thiazolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, furyl,
indolyl, quinolyl, isoquinolyl, benzofuryl, benzothienyl,
benzimidazolyl, benzopyrazolyl, benzoxazolyl, benzoisooxazolyl, and
benzothiazolyl.
4. The compound or the pharmaceutically acceptable salt thereof
according to any one of claims 1-3, wherein the aryl is selected
from the group consisting of phenyl, naphthyl, and anthryl.
5. The compound or the pharmaceutically acceptable salt thereof
according to any one of claims 1-3, wherein the alkyl is selected
from the group consisting of methyl, ethyl, propyl, iso-propyl,
n-butyl, iso-butyl, tert-butyl, pentyl, iso-pentyl, neo-pentyl, and
hexyl; the alkenyl is selected from the group consisting of vinyl,
propenyl, butenyl, pentenyl, and hexenyl; the alkynyl is selected
from the group consisting of ethynyl, propynyl, butynyl, pentynyl,
and hexynyl.
6. The compound or the pharmaceutically acceptable salt thereof
according to claim 4, wherein the alkyl is selected from the group
consisting of methyl, ethyl, propyl, iso-propyl, n-butyl,
iso-butyl, tert-butyl, pentyl, iso-pentyl, neo-pentyl, and hexyl;
the alkenyl is selected from the group consisting of vinyl,
propenyl, butenyl, pentenyl, and hexenyl; the alkynyl is selected
from the group consisting of ethynyl, propynyl, butynyl, pentynyl,
and hexynyl.
7. The compound or the pharmaceutically acceptable salt thereof
according to claim 6, wherein the compound is: ##STR00002##
8. The compound or the pharmaceutically acceptable salt thereof
according to any one of claims 1-3, wherein the pharmaceutically
acceptable salt is selected from the group consisting of an organic
acid salt, an inorganic acid salt, an alkali metal salt, an alkali
earth metal salt, and an inner salt of the compound.
9. A pharmaceutical composition comprising a compound of any one of
claims 1-3 and optionally a pharmaceutically acceptable
carrier.
10. A method of treating sequelae of ischemic cerebral stroke,
comprising administering a therapeutically effective amount of the
compound of any one of claims 1-3.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of International Patent
Application No. PCT/CN2015/087048, filed Aug. 14, 2015, which is
incorporated by reference herein in its entirety.
TECHNICAL FIELD
[0002] The present invention relates to a medicinal compound for
treating sequelae of ischemic cerebral stroke and the use of the
same in the manufacture of a medicament for treating sequelae of
ischemic cerebral stroke.
BACKGROUND ART
[0003] Cerebral stroke is a disease in which a cerebral tissue
damage is caused by a sudden blood vessel rupture or by a blood
circulation disturbance arising from vascular obstruction in
brain.
[0004] Ischemic cerebral stroke means necrosis of local cerebral
tissues including neurocytes, neurogliocytes, and blood vessel due
to deficient in blood supply. The fundamental cause of the ischemic
cerebral stroke is that the extracranial or intracranial arteries
leading to the brain have occlusive lesions and fail to achieve
prompt and adequate collateral circulation so that the metabolism
requirement of the local cerebral tissues cannot be met by the
available blood supply. The incidence of ischemic cerebral stroke
is higher than that of hemorrhagic cerebral stroke and accounts for
60%-70% of all cerebral stroke cases. The occlusion and stenosis in
internal carotid and vertebral arteries may cause ischemic cerebral
stroke in people older than 40 in a higher frequency in males than
in females. Some severe cases result in death.
[0005] The sequelae of ischemic cerebral stroke means the symptoms
of hemiplegia, speech disorder, or facial paralysis and the like
occurred after the ischemic cerebral stroke, collectively referred
to as sequelae of ischemic cerebral stroke. Mainly, these include
the following symptoms: semiplegia (hemiplegia), hemi-limb
disabilities, numbness of limbs, hemianopsia, aphasia, or crossed
hemiplegia, contralateral sensory disturbance, external
ophthalmoplegia, nystagmus, dyslalia, speech disorder, memory
deterioration, facial paralysis, dysphagia, choking on food and
drink, dystaxia, dizzy and headache, and the like.
[0006] Reckoning on the survey data obtained 20 years ago, there
are about 2 million of new cerebral stroke cases every year in
China, more than 150 million persons died from cerebral stroke
every year, and there are about 600-700 million survived from
cerebral stroke. Among the survival patients, about 75%-80% would
have different levels of sequelae, with over 40% of severe
disablility. About 1/4 to 1/3 of them may relapse within 2 to 5
years. With the improvement in living standards and the change of
the life style in recent years, the population incidence of
cerebral stroke is still rising in China. In particular, because of
the improvement of medical conditions and the advancement of
clinical technology, the mortality rate of stroke significantly
declined while, however, leading to a great increase in the
morbidity of the sequelae of cerebral stroke, which causes a
greater disease burden. The cerebral stroke has become a leading
disease that jeopardizes the health of middle aged and elderly
people in China.
[0007] At present, butylphthalide (NBP) is a drug approved for the
treatment of sequelae of ischemic cerebral stroke. The sodium
chloride injection of butylphthalide (which has the principal
component of butylphthalide with a chemical name of
dl-3-n-butylphthalide (called as butylphthalide or NBP for short))
is used for improving neurological function deficits in a patient
with acute ischemic cerebral stroke. The drug is administered
through an intravenous drip within 48 hours from the onset of the
disease, twice per day, each 25 mg (100 ml) over a period of less
than 50 minutes with a time interval of not less than 6 hours
between two administrations during a treatment course of 14 days.
Since a PVC infusion device has a significant absorption effect on
butylphthalide, a PE infusion device can only be used for the
infusion of the butylphthalide product. There is no research data
about the efficacy or safety of the drug administered 48 hours
after the onset of the disease. The butylphthalide soft capsule has
an indication of mild or moderate acute ischemic cerebral stroke.
In a specification of 0.1 g, the drug is administered orally under
fasting, two capsules (0.2 g) every time, four times every day
during a treatment course of 10-12 days, or as recommended by the
doctor. Unwanted effects include mild transaminase elevation, which
can be normalized after drug withdrawal according to some follow-up
cases. Nausea, abdominal discomfort, rash, psychiatric symptoms and
the like happen occasionally.
[0008] In addition, aspirin is also used to treat sequelae of
ischemic cerebral stroke. Aspirin has a generic name of
acetylsalicylic acid, and is an antipyretic analgesics with a long
history since its discovery in 1899. It has been used for treating
cold, fever, headache, toothache, arthralgia, and rheumatism, and
is capable of inhibiting platelet aggregation, thereby preventing
and treating ischemic cardiopathy, angina, cardiopulmonary
infarction, and cerebral thrombosis. However, aspirin has no
significant efficacy on the treatment of sequelae of ischemic
cerebral stoke, but has multiple adverse effects. Accordingly, when
aspirin is used to treat various diseases, one should keep close
watch over its adverse effects.
[0009] Therefore, there still is a great need now to develop a new
medicinal substance for treating sequelae of ischemic cerebral
stroke in the current society.
SUMMARY OF THE INVENTION
[0010] It is one objective of the present invention to provide a
compound represented by the following formula (I) or the
pharmaceutically acceptable salt thereof for treating sequelae of
ischemic cerebral stroke:
H--(NH--CHR.sub.1--CO)--(NH--CHR.sub.2--CO)--(NH--CHR.sub.3--CO)--(NH--C-
HR.sub.4--CO)--(NH--CHR.sub.5--CO)--(NH--CHR.sub.6--CO)--(NH--CHR.sub.7--C-
O)--OH (I)
wherein each of R.sub.1-R.sub.7 is independently selected from the
group consisting of alkyl, alkenyl, alkynyl, alkoxyl, or alkylthio
which are optionally substituted by one or more substituents
selected from the group consisting of halo, hydroxyl, sulfydryl,
carboxyl, amino, nitro, cyano, carbamoyl, optionally substituted
cycloalkyl, optionally substituted aryl, and optionally substituted
heteroaryl, wherein the substituents in the optionally substituted
cycloalkyl, optionally substituted aryl and optionally substituted
heteroaryl are one or more selected from the group consisting of
hydroxyl, sulfydryl, amino, nitro, cyano, and carboxyl.
[0011] Preferably, the present invention provides a compound of
formula (I):
H--(NH--CHR.sub.1--CO)--(NH--CHR.sub.2--CO)--(NH--CHR.sub.3--CO)--(NH--C-
HR.sub.4--CO)--(NH--CHR.sub.5--CO)--(NH--CHR.sub.6--CO)--(NH--CHR.sub.7--C-
O)--OH (I) [0012] wherein each of R.sub.1, R.sub.4 and R.sub.6 is
independently selected from the group consisting of C.sub.1-C.sub.6
alkyl, C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl; [0013]
R.sub.2 is C.sub.1-C.sub.6 alkyl which is substituted with five to
ten membered heteroaryl optionally substituted with halo, hydroxyl,
sulfydryl, carboxyl, amino, nitro, or cyano; [0014] R.sub.3 is
C.sub.1-C.sub.6 alkyl group substituted by halo, hydroxyl,
sulfydryl, or amino; [0015] R.sub.5 is C.sub.1-C.sub.6 alkyl
substituted with carbamoyl; and [0016] R.sub.7 is C.sub.1-C.sub.6
alkyl which is substituted with C6-C14 aryl optionally substituted
with halo, hydroxyl, sulfydryl, carboxyl, amino, nitro, or cyano;
[0017] or a pharmaceutically acceptable salt thereof.
[0018] In one aspect, the present invention provides a method for
preparing a compound of formula (I), which links multiple monomeric
units together through a condensation reaction using a conventional
polymerization method in the art, in particular a solid phase
synthesis method and results in the desired compound.
[0019] In one aspect, the present invention provides a
pharmaceutical composition comprising the compound of formula (I).
The pharmaceutical composition comprises at least one compound
according to the invention and optionally a pharmaceutically
acceptable carrier.
[0020] In one aspect, the present invention also provides the use
of the compound according to the invention or a pharmaceutical
composition comprising the same in the manufacture of a medicament
for treating sequelae of ischemic cerebral stroke.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] FIG. 1 illustrates the weight conditions of rats at
different times (observed until 22 weeks) in a rat 1.5 h
ischemia-reperfusion model.
[0022] FIG. 2 illustrates the neurologic deficit scores of rats at
different times (observed until 22 weeks) in a rat 1.5 h
ischemia-reperfusion model.
[0023] FIG. 3 illustrates the therapeutic effects of compound of
formula (III) on rats continuously administered for 4 weeks after
1.5 h ischemia and one week reperfusion (observed until 22
weeks)--weight condition.
[0024] FIG. 4 illustrates the therapeutic effects of compound of
formula (III) on rats continuously administered for 4 weeks after
1.5 h ischemia and one week reperfusion (observed until 22
weeks)--neurologic deficit score.
[0025] FIG. 5 illustrates the therapeutic effects of compound of
formula (III) on rats continuously administered for 4 weeks after
1.5 h ischemia and two week reperfusion (observed until 7
weeks)--weight condition.
[0026] FIG. 6 illustrates the therapeutic effects of compound of
formula (III) on rats continuously administered for 4 weeks after
1.5 h ischemia and two week reperfusion (observed until 7
weeks)--neurologic deficit score.
[0027] FIG. 7 illustrates the measurement results of cerebral
infarction injury rate in rats (treated with continuous
administration for 4 weeks after 1.5 h ischemia and two week
reperfusion).
[0028] FIG. 8 illustrates the measurement results of cerebral
infarction injury rate in rats (treated with continuous
administration for 4 weeks after 1.5 h ischemia and one week
reperfusion)
DETAILED DESCRIPTION
Definitions
[0029] As used herein, the term "alkyl" refers to a straight or
branched chain alkane hydrocarbon group containing a specified
number of carbon atoms, such as C.sub.1-C.sub.18 alkyl,
C.sub.1-C.sub.12 alkyl, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.4
alkyl and the like. Examples of alkyl include, but are not limited
to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,
tert-butyl, pentyl, iso-pentyl, neo-pentyl, hexyl, heptyl, octyl,
nonyl, decyl and the like.
[0030] The term "alkenyl" refers to a straight or branched chain
alkene hydrocarbon group containing a specified number of carbon
atoms, such as C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.4 alkenyl and the like. Examples of alkenyl include,
but are not limited to, vinyl, propenyl, butenyl, pentenyl,
hexenyl, heptenyl, octenyl, nonenyl, decenyl and the like.
[0031] The term "alkynyl" refers to a straight or branched alkyne
hydrocarbon group containing a specified number of carbon atoms,
such as C.sub.2-C.sub.12 alkynyl, C.sub.2-C.sub.6 alkynyl,
C.sub.2-C.sub.4 alkynyl and the like. Examples of alkynyl include,
but are not limited to, ethynyl, propynyl, butynyl, pentynyl,
hexynyl, heptynyl, octynyl, nonenyl, decynyl, and the like.
[0032] The term "cycloalkyl" refers to a hydrocarbon group of
saturated 3-8 membered monocyclic system. Specific examples of
cycloalkyl include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and
the like.
[0033] The term "aryl" refers to a monocyclic aromatic group or a
condensed or non-condensed polycyclic aromatic group containing
6-14 carbon atoms, preferably 6-10 carbon atoms. Examples of aryl
preferably are, but are not limited to, phenyl, bi-phenyl,
naphthyl, anthryl, 5,6,7,8-tetrahydronaphthyl,
2,3-dihydrobenzofuryl, and the like.
[0034] The term "heteroaryl" refers to a five to ten membered
aromatic cyclic group containing 1-4 heteroatoms selected from
nitrogen, sulfur or oxygen as ring atoms. The heteroaryl may be a
monocyclic heteroaryl group containing 5-6 ring atoms, or a
bicyclic heteroaryl group containing 7-10 ring atoms. Examples of
the heteroaryl include, but are not limited to, pyrrolyl,
pyrazolyl, imidazolyl, oxazolyl, thienyl, iso-oxazolyl,
oxadiazolyl, thiazolyl, triazolyl, tetrazolyl, pyridinyl,
pyrimidinyl, furyl, indolyl, quinolyl, isoquinolyl, benzofuryl,
benzothienyl, benzimidazolyl, benzopyrazolyl, benzoxazolyl,
benzoisooxazolyl, benzothiazolyl and the like.
[0035] The term "halo" refers to --F, --Cl, --Br, or --I.
[0036] The term "alkoxyl" refers to --O-alkyl.
[0037] The term "alkylthio" refers to --S-alkyl.
[0038] The term "pharmaceutically acceptable salt" includes, but is
not limited to, organic acid salts, inorganic acid salts, metal
salts, ammonium salts and inner salts of the compounds according to
the present invention, among which the organic acid salts include,
but are not limited to, tosilate, mesylate, malate, acetate,
citrate, lactate, ascorbate, tartrate, succinate, fumarate,
maleate, oxalate, malonate, and the like; the inorganic acid salts
include, but are not limited to, hydrochloride, sulphate,
phosphate, nitrate, hydrobromide, hydroiodate, sulfite, carbonate,
bicarbonate, bisulphate, dihydrophosphate, hydrophosphate, and the
like; and the metal salts include, but are not limited to, sodium
salts, potassium salts, lithium salts, magnesium salts, calcium
salts, ferric salts, and the like.
[0039] The term "pharmaceutically acceptable carrier" includes
conventional excipients, diluents, disintegrants, binders,
lubricants, sweeteners, stabilizers, solubilizers, preservatives,
and the like.
[0040] The term "prevention" refers to completely or partly
preventing the occurrence of a disease or the symptoms thereof.
[0041] The term "treat" refers to completely or partly alleviating
or curing a disease or the symptoms thereof, comprising (a)
alleviating the symptoms of the disease and (b) eliminating the
symptoms of the disease.
[0042] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt provided herein, each of
R.sub.1-R.sub.7 is independently selected from the group consisting
of optionally substituted C.sub.1-C.sub.18 alkyl (preferably
C.sub.1-C.sub.12 alkyl, more preferably C.sub.1-C.sub.6 alkyl, more
preferably methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl,
tert-butyl, pentyl, hexyl), optionally substituted C.sub.2-C.sub.12
alkenyl (preferably C.sub.2-C.sub.6 alkenyl, more preferably
C.sub.2-C.sub.4 alkenyl, more preferably vinyl, propenyl, butenyl),
optionally substituted C.sub.2-C.sub.12 alkynyl (preferably
C.sub.2-C.sub.6 alkynyl, more preferably C.sub.2-C.sub.4 alkynyl,
more preferably ethynyl, propynyl, butynyl), optionally substituted
C.sub.1-C.sub.18 alkoxyl (preferably C.sub.1-C.sub.12 alkoxyl, more
preferably C.sub.1-C.sub.6 alkoxyl, more preferably methoxyl,
ethoxyl, propoxyl, butoxyl, pentyloxyl, hexyloxyl), and optionally
substituted C.sub.1-C.sub.18 alkylthio (preferably C.sub.1-C.sub.12
alkylthio, more preferably C.sub.1-C.sub.6 alkylthio, more
preferably methylthio, ethylthio, propylthio, butylthio,
pentylthio, hexylthio), wherein the substituents are one or more
selected from the group consisting of halo, hydroxyl, sulfydryl,
carboxyl, amino, nitro, cyano, carbamoyl, optionally substituted
C.sub.3-C.sub.8 cycloalkyl, optionally substituted C.sub.6-C.sub.14
aryl and optionally substituted five to ten membered heteroaryl;
and in the optionally substituted C.sub.3-C.sub.8 cycloalkyl,
optionally substituted C.sub.6-C.sub.14 aryl and optionally
substituted C.sub.6-C.sub.10 heteroaryl, the substituents are one
or more selected from the group consisting of hydroxyl, sulfydryl,
amino, nitro, cyano, and carboxyl.
[0043] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, each of
R.sub.1-R.sub.7 is independently selected from the group consisting
of optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted C.sub.2-C.sub.6 alkenyl, optionally substituted
C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.1-C.sub.6
alkoxyl, and optionally substituted C.sub.1-C.sub.6 alkylthio,
wherein the substituents are one or more selected from the group
consisting of halo, hydroxyl, sulfydryl, carboxyl, amino, nitro,
cyano, and carbamoyl, optionally substituted C.sub.3-C.sub.8
cycloalkyl, optionally substituted C.sub.6-C.sub.14 aryl and
optionally substituted five to ten membered heteroaryl, and in the
optionally substituted C.sub.3-C.sub.8 cycloalkyl, optionally
substituted C.sub.6-C.sub.14 aryl and optionally substituted five
to ten membered heteroaryl, the substituents are one or more
selected from the group consisting of hydroxyl, sulfydryl, amino,
nitro, cyano, and carboxyl.
[0044] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, each of
R.sub.1-R.sub.7 is independently selected from the group consisting
of optionally substituted C.sub.1-C.sub.6 alkyl, optionally
substituted C.sub.2-C.sub.6 alkenyl, optionally substituted
C.sub.2-C.sub.6 alkynyl, optionally substituted C.sub.1-C.sub.6
alkoxyl, and optionally substituted C.sub.1-C.sub.6 alkylthio; and
wherein R.sub.2 is optionally substituted with five to ten membered
heteroaryl; R.sub.3 is optionally substituted with halo, hydroxyl,
sulfydryl, carboxyl, amino, nitro, or cyano; R.sub.5 is optionally
substituted with carboxyl, or carbamoyl; and R.sub.7 is optionally
substituted with C.sub.6-C.sub.14 aryl optionally substituted with
halo, hydroxyl, sulfydryl, carboxyl, amino, nitro, or cyano.
[0045] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, the
C.sub.6-C.sub.14 aryl is selected from the group consisting of
phenyl, naphthyl, anthryl, and the like; the five to ten membered
heteroaryl is selected from the group consisting of pyrrolyl,
pyrazolyl, imidazolyl, oxazolyl, thienyl, iso-oxazolyl,
oxadiazolyl, thiazolyl, triazolyl, tetrazolyl, pyridinyl,
pyrimidinyl, furyl, indolyl, quinolyl, isoquinolyl, benzofuryl,
benzo thienyl, benzimidazolyl, benzopyrazolyl, benzoxazolyl, benzo
isooxazolyl, benzothiazolyl and the like.
[0046] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, the
heteroaryl is selected from five to six membered heteroaryl.
[0047] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, the
heteroaryl is selected from the group consisting of pyrrolyl,
pyrazolyl, imidazolyl, oxazolyl, thienyl, iso-oxazolyl,
oxadiazolyl, thiazolyl, triazolyl, tetrazolyl, pyridinyl,
pyrimidinyl, furyl, indolyl, quinolyl, isoquinolyl, benzofuryl,
benzothienyl, benzimidazolyl, benzopyrazolyl, benzoxazolyl,
benzoisooxazolyl, and benzothiazolyl.
[0048] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, the aryl
is selected from the group consisting of phenyl, naphthyl, and
anthryl.
[0049] Preferably, in the compound of formula (I) or a
pharmaceutically acceptable salt thereof provided herein, the alkyl
is selected from the group consisting of methyl, ethyl, propyl,
iso-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, iso-pentyl,
neo-pentyl, and hexyl; the alkenyl is selected from the group
consisting of vinyl, propenyl, butenyl, pentenyl, and hexenyl; the
alkynyl is selected from the group consisting of ethynyl, propynyl,
butynyl, pentynyl, and hexynyl.
[0050] In a preferable embodiment of the present invention, the
compound according to the invention has the following
structure:
##STR00001##
[0051] The compounds according to the present invention can be
prepared by a conventional organic chemical synthesis method.
Specifically, the compounds according to the present invention can
be prepared by a solid phase synthesis method commonly used in the
art, wherein a polymeric resin is used as an insoluble solid phase
support. Firstly, a first amino acid protected at its N-terminal
(e.g. with a protective group of Fmoc) is covalently linked to the
solid phase support. Under a basic condition, the amino group is
deprotected. Then a condensation reaction between the carboxyl of a
second amino acid likely protected at its N-terminal and the amino
of the first amino acid linked to the solid phase support is
carried out to form a peptide bond. Through the similar steps,
other amino acids are further linked to the amino terminal of the
second amino acid until the desired peptide chain backbone is
formed. Then, the peptide chain is separated from the resin under
suitable reaction conditions and other protecting groups remained
in the peptide chain are removed. And final products are obtained
through purification.
[0052] The present invention also provides a pharmaceutical
composition comprising the compound according to the present
invention, which comprises the compound as described herein and
optionally a pharmaceutically acceptable carrier. To prepare a
pharmaceutical composition comprising the compound described
herein, the pharmaceutically acceptable carrier may be solid or
liquid. Among others, the solid carrier may be one or more
materials used as excipients, diluents, sweeteners, solubilizers,
lubricants, binders, tablet disintegrants, stabilizers,
preservatives, or encapsulating materials. The liquid carrier may
be solvents or liquid dispersion media. Suitable solid carriers
include, but are not limited to, for example, cellulose, glucose,
lactose, mannitol, magnesium stearate, magnesium carbonate,
saccharin sodium, sucrose, dextrin, talc, starch, pectin, gelatin,
tragacanth, arabic gum, sodium alginate, parabens, methylcellulose,
sodium carboxymethyl cellulose, a low-melting point wax, cocoa
butter, and the like. Suitable liquid carriers include, but are not
limited to, water, ethanol, polylol (such as glycerol, propanediol,
liquid polyethylene glycol, etc), a vegetable oil, glyceride and a
mixture thereof.
[0053] The pharmaceutical composition according to the present
invention may be prepared by a known method, including conventional
blending, granulating, tableting, coating, dissolving, or
lyophilization processes.
[0054] The pharmaceutical composition according to the present
invention may be administered to the patients via various routes,
such as oral, local (such as topical), systemic, intravenous,
intramuscular, or mucosal.
[0055] Depending on the administration route, the pharmaceutical
composition according to the present invention may be prepared into
various dosage forms conventional in the art, such as tablet,
capsule, pill, emulsion, injection, pulvis, granule, ointment,
patch, powder injection, suspension, cream, aerosol, drop, lozenge,
and the like.
[0056] In a tablet, an active ingredient can be blended with a
carrier having required binding capacity in an appropriate ratio
and pressed into a desired shape and size. In a powder dosage form,
a carrier is finely divided solids blended with a finely divided
active ingredient. A powder and tablet generally contains about 5
or 10% to about 70% of active ingredient. Suitable carriers
include, for example, magnesium carbonate, magnesium stearate,
talc, sugars, lactose, pectin, dextrin, starch, gelatin,
tragacanth, methyl cellulose, sodium carboxymethylcellulose, a
low-melting-point wax, cocoa butter and the like.
[0057] The compound described herein can be formulated for
parenteral administration (e.g. through injection) and may be
present in a unit dosage form in an ampoule, a pre-filled syringe,
a small-volume infusion bottle, or in a multiple dose container.
The composition may be in the form of suspensions, solutions, or
emulsions in an oily or aqueous medium. Alternatively, the active
ingredients may be in the form of powders before use, which is
obtained by sterile separation of sterile solids or lyophilization
of solutions, and may be reconstituted in a suitable medium such as
sterile and pyrogen-free water.
[0058] An aqueous solution suitable for oral use can be prepared by
dissolving the active ingredient into water and adding suitable
colorants, sweeteners, preservatives, stabilizers and/or
solubilizers as needed.
[0059] An aqueous suspension suitable for oral use can be prepared
by dispersing the finely divided active ingredient into water with
viscous materials such as a natural or synthetic gum, resin,
methylcellulose, sodium carboxylmethylcellulose, or other
well-known suspending agents.
[0060] The compound described herein can be formulated into a form
of ointment, cream, or transdermal patch for local administration
to epidermis. The ointment or cream can be formulated with, for
example, an aqueous or oily matrix added with a suitable thicker
and/or gelling agent.
[0061] The administration to respiratory tract can be implemented
using an aerosol formulation, wherein the active ingredient is
provided in a pressurized package having suitable propellants (such
as chlorofluorocarbon (e.g. dichlorodifluoromethane,
trichlorofluoromethane or dichlorotetrafluoroethane), carbon
dioxide or other suitable gases). The aerosol can conveniently
comprise surfactants (such as lecithin). The drug dosage can be
controlled by a metering valve.
[0062] In addition, if needed, a formulation suitable for sustained
release, delayed release, or retarded release may also be used.
[0063] The pharmaceutical formulation is preferably in a unit
dosage form, in which the formulation is subdivided into unit
dosages containing a suitable amount of active ingredient. The unit
dosage form can be packed in a package containing discrete
quantities of the formulation, such as packaged tablets, capsules,
or powders in vials or ampoules
[0064] The present invention also provides use of the compound or
pharmaceutical composition according to the present invention in
the manufacture of a medicament, in particular a medicament for
treating sequelae of ischemic cerebral stroke. Accordingly, the
present invention provides a method for treating sequelae of
ischemic cerebral stroke, comprising administering a
therapeutically effective amount of the pharmaceutical composition
comprising at least one compound according to the present invention
or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carriers to a patient in need thereof.
In another aspect, the present invention provides a method for
treating sequelae of ischemic cerebral stroke, comprising
administering a therapeutically effective amount of at least one
compound according to the present invention or a pharmaceutically
acceptable salt thereof to a patient in need thereof. In another
aspect, the present application provides the compound of the
present application for the use of treating sequelae of ischemic
cerebral stroke. The precise dosage required is determined based on
the attendant physician's judgment. Generally, the dosage of the
active compound to be administered may be, for example, about 0.1
to about 100 mg per day, about 0.1 to about 75 mg/day, about 0.1 to
about 50 mg/day, or about 5 to about 10 mg/day. The desired dosage
depends on the specific compound used, severity of disease,
administration route, weight and healthy condition of the patient
as well as the attendant physician's judgment.
EXAMPLES
[0065] The present invention will be illustrated in more details in
the following examples. However, it should be understood that the
following examples intend to illustrate the present invention, but
do not limit the scope of the present invention in any way.
Preparation of the Compound
Example 1
Preparation of the Compound of Formula (III)
1. Solid Phase Synthesis of Peptide Chain
[0066] 100 g of dichloro-tritylchloride resin (having a degree of
substitution of 1 mmol/g resin) was added into a 5 L reaction
flask, to which 1 L dichloromethane was added. After 10 minutes,
the resin sufficiently swelled in the solution, and 31.08 g of
Fmoc-Tyr(tBu)-OH were added followed by 38 ml of DIEA. The reaction
was continued for 30 minutes. Then, 1500 ml of methanol was added
to terminate the reaction. The resin was filtered off, washed
successively with isopropanol (IPA), DMF, isopropanol, DMF,
isopropanol and ethyl ether, and dried in a fume hood to constant
weight.
[0067] 100 g of dried resin carrying Fmoc-Tyr(tBu)-OH was added
into a 2 L reaction flask, and 1 L of DMF was added so that the
resin sufficiently swelled in the solvent of DMF. DMF was drawn
under a negative pressure into a waste liquid bottle, and 1 L of
piperidine/DMF solution (25%) was added. The reaction flask was
placed on a shaker at a speed of 120 rpm and the reaction was
conducted for 30 minutes. Then, the deprotection solution was drawn
under a negative pressure into a waste liquid bottle. The resin is
washed successively with DMF, isopropanol, DMF, isopropanol and
DMF, and all washings were transferred under a negative pressure to
a waste liquid bottle.
[0068] 66 g of Fmoc-Leu-OH, 30 ml of DIC, and 27 g of HOBt were
added into a clean conical flask and were dissolved in 500 ml of
DMF. After being activated on a shaker for 10 minutes, the
activated solution was poured into the previous reaction flask. The
reaction flask was placed on a shaker at a speed of 120 rpm for 120
minutes. After the reaction was completed, the reaction solution
was drawn under a negative pressure into a waste liquid bottle. The
resin was successively washed with DMF, isopropanol, DMF and
isopropanol, and al washings were transferred under a negative
pressure to a waste liquid bottle.
[0069] Following the above procedure, the protected amino acids of
Fmoc-Gln(Trt)-OH, Fmoc-Val-OH, Fmoc-Ser(tBu)-OH, Fmoc-His(Trt)-OH
and Fmoc-Val-OH were linked in sequence to the
resin-Tyr(tBu)-Leu-OH, forming
resin-Tyr(tBu)-Leu-Gln(Trt)-Val-Ser(tBu)-His(Trt)-Val-NH.sub.2. The
specific reaction reagents and the amounts used as well as the
specific reaction parameters are shown in the following table
(based on 100 g resin):
TABLE-US-00001 Coupling sequence Deprotection Washing Activation
Washing 1 piperidine/DMF DMF (1 L) Fmoc-Gln(Trt)-OH DMF (1 L)
(1:4), 30 min IPA (1 L) 121 g, DIC 30 ml, IPA (1 L) 3 cycles HOBt
27 g 3 cycles 2 piperidine/DMF DMF (1 L) Fmoc-Val-OH 67.2 g, DMF (1
L) (1:4), 30 min IPA (1 L) DIC 30 ml, IPA (1 L) 3 cycles HOBt 27 g
3 cycles 3 piperidine/DMF DMF (1 L) Fmoc-Ser(tBu)-OH DMF (1 L)
(1:4), 30 min IPA (1 L) 76 g, DIC 30 ml IPA (1 L) 3 cycles HOBt 27
g 3 cycles 4 piperidine/DMF DMF (1 L) Fmoc-His(Trt)-OH DMF (1 L)
(1:4), 30 min IPA (1 L) 122.8 g, DIC 30 ml IPA (1 L) 3 cycles HOBt
27 g 3 cycles 5 piperidine/DMF DMF (1 L) Fmoc-Val-OH 67.2 g, DMF (1
L) (1:4), 30 min IPA (1 L) DIC 30 ml IPA (1 L) 3 cycles HOBt 27 g 3
cycles
2. Cleave of the Resin and Release of the Peptide Chain
[0070] The resin obtained after final washing was dried in a fume
hood, and then was transferred to a 2 L round-bottom flask. After
the addition of 1200 ml of TFA/H.sub.2O (95:5), the round-bottom
flask was placed on a shaker at 20 rpm to react for 2 hours. Then,
the reaction solution was filtered through sand cores, and the
filtrate was added in batches into 3 L of anhydrous ethyl ether. At
this point, a large amount of white precipitates separated out.
After standing for a while to full precipitation, filtering through
a Buchner funnel, washing with ethyl ether for three times, and
weighting to give a crude product of the compound of formula
(III).
3. Purification of the Crude Product
[0071] The resulting crude product of the compound of formula (III)
was dissolved in purified water at a concentration of 25 mg/ml, and
centrifuged. The undissolved part of the sample was discarded. Each
160 ml of the sample solution was loaded onto a HPLC dynamic axial
compression column (ID 150 mm). Elution was carried out following
gradient schemes for separation:
TABLE-US-00002 Water Acetonitrile (containing 0.1% of (containing
0.1% Time (min) Flow rate trifluoroacetic acid) of trifluoroacetic
acid) 0 430 14% 86% 10 430 14% 86% 70 430 26% 74%
[0072] Analytic HPLC was used to analyze the purity of each
component. The samples having a purity of more than 98.5% were
combined. The combined sample was evaporated on a rotary evaporator
in a water bath of 40 Celsius degree and concentrated to a
concentration of 10 mg/ml, and after being placed in a refrigerator
to freeze, a pure product of the compound of formula (III) was
obtained in a purity of more than 98%. LC-MS 845.42
[M+H].sup.+.
Biological Assessment
1. Materials
(1) Primary Reagents and Drugs
TABLE-US-00003 [0073] Test compounds self-made Chloral hydrate
Sinopharm Chemical Reagent Co., Ltd Red tetrazoline (TTC) Sinopharm
Chemical Reagent Co., Ltd Paraformaldehyde Shanghai Lingfeng
Chemical Reagent Co., Ltd
(2) Experimental Animals
[0074] Sprague-Dawley (SD) rats, male, weighing 280-320 g, clean,
feeding conditions: room temperature of 20-25.degree. C., humidity
of 30-60%, well ventilated, ad libitum diet; the animals were
acclimatized in the feeding environment for three days prior to the
experiment.
2. Establishment of the Right Cerebral Artery Ischemia/Reperfusion
Model in Rats.
[0075] The rats were anesthetized with 4% chloral hydrate (350
mg/kg, ip.) and then were fixed in a supine position. A midline
ventral neck incision was made, the right common carotid artery was
separated and snared with two sutures for further use. Then, the
internal carotid artery and external carotid artery were separated,
and the external carotid artery was ligatured. The separated common
carotid artery was ligatured at its proximal end with a thread, and
the blood flow of the same was blocked at its distal end with a
bulldog clamp. A small opening was cut therebetween and a nylon
suture (4-0) which is heated at one end into a bead-like shape
(having a diameter of <0.3 mm) was inserted into the small
opening. The bulldog clamp was removed, and the nylon suture was
gently advanced to the origin of the anterior cerebral artery
(18-20 mm), thereby causing arterial ischemia in brain by blocking
the blood supply of the cerebral artery. The skin was sutured and
was sterilized with tincture of iodine. After 4 hours or 2 hours or
1.5 hours, the initial traumatic surface was open again by cutting
and the nylon suture was withdrawn slowly from the common carotid
artery which was ligatured with a suture and fixed. The skin was
sutured again and was sterilized with tincture of iodine.
3. Assay of Cerebral Infarction Injury Rate in Rats
[0076] At different time after the operation, the animals were
sacrificed and the brains were collected, from which the bulbus
olfactorius, opisthencephalon and low brain stem were removed. The
brains were cut four times backward in a coronal plane using a die
to divide it into five slices each having a thickness of 3 mm,
wherein the first cut was made at the middle of the connecting line
of brain anterior pole and optic chiasma, the second cut was made
at the site of optic chiasma, the third cut was made at infundbular
stalk, and the fourth cut was made between infundbular stalk and
posterior pole of posterior pituitary. The slices were stained with
2% red tetrazolium (TTC): 1.5 ml of 2% TTC, 3.4 ml of saline, and
incubated in dark at 37.degree. C. for 30 min. Normal tissues
appeared in red, while infarct tissues appeared in white or showed
stroke capsules which are defected when slicing. The five brain
slices were arranged in original order and photographed. The
percentage of cerebral infarction injury was calculated as a volume
(area) percentage of the remaining volume (area) in the infarct
cerebral hemisphere to the volume (area) of the contralateral
cerebral hemisphere.
4. Preparation of Test Samples
[0077] The compound of formula (III) was dissolved in PBS subjected
to high temperature sterilization at 121.degree. C. to formulate
various solutions of 0.1-9.6 mg/ml, which was filtered through a
millipore filter membrane of 22 .mu.m and stored in a refrigerator
at 4.degree. C. until use. The entire preparation process was
carried out in a super clean bench.
5. Administration Method
[0078] From the beginning of the administration, various doses of
0.1-9.6 mg/kg were injected intraperitoneally into respective
animal groups once a day.
6. Neurological Score
[0079] Neurologic deficit scores were assessed for the above
experimental animals according to the following procedure before
the beginning of experiment and 24 hours, 1 day, 3 days, 7 days, 14
days, 28 days, 56 days, and 112 days after the beginning of
experiment; Score rules: see Zea Longer score and NSS score, full
score of 10;
(i) Lifting an Animal by Tail: (Full Score of 1)
[0080] Fore limbs flexing 1
(ii) Observation of Animals Placed on the Ground: (Full Score of
4)
[0081] Walking normally with physical agility 0 Acting slowly,
being unable to walk straight, and failing to stretch contralateral
limbs fully 1 Circling toward the contralateral side 2 Falling
toward the contralateral side or falling down when walking 3 Being
unable to walk spontaneously accompanied by severe loss of
consciousness 4 (iii) Observations of Animals Placed on a Balance
Beam: (Full Score of 4) Walking evenly, stably, and promptly 0 The
animals could walk, but one limb slid down or turned around on the
balance beam for a long time 1 The animals could not walk smoothly,
two hind legs slid down, but they could stay on the balance beam
stably without falling 2 The animals tried to grab the balance
beam, but still fell 3 The animals did not try to grab the balance
beam, and directly fell 4
(iv) Observing the Animal's Condition: (Full Score of 1)
[0082] Homer syndrome appeared, the head skews toward one side, one
eye lid drooped, and there was no obvious bleeding around the eyes
1
Example 2
[0083] Normal male SD rats, weighing 280.about.320 g, were used in
six experiments on 1.5-hour ischemia/reperfusion model groups
(I1.5R+ model). Monitoring the scores of neuroethology symptoms 24
hours after the cerebral ischemia reperfusion injury and at each
subsequent week in rats of the 1.5-hours ischemia/reperfusion model
groups. The final results and sample size: results from the I1.5R+
model group in six experiments were combined (animal data: 31
animals in total in the I1.5R+ model groups. In addition, the data
of first week was the combined data from the experimental groups
and the model group, the animal number in the 24-hour data was 101,
and the animal number in the first week data was 106).
[0084] The weights and scores of SD rats in the 1.5-hour
ischemia/reperfusion model were recorded at different time points
later. The results showed that the weights of SD rats in the
1.5-hour ischemia/reperfusion showed a tendency of gradually
rising, and the neurological score of 24-hour was slightly greater
than 4 and decreased down to 3 at one week, then the score
fluctuated around 3 for a long time. The detailed results were
shown in FIGS. 1 and 2. The assay of cerebral infarction injury
ratio in rats was seen in FIGS. 7 and 8.
Example 3
[0085] Normal male SD rats, weighing 280.about.320 g, were used in
three experiments. In the first experiment, animals were divided
randomly into two groups, one group in which the administration
started one week after the 1.5-hour ischemia/reperfusion and
continued for four weeks (I1.5R+1 week+dosing for 4 weeks), and the
other group for the 1.5-hour ischemia/reperfusion model without
administering the compound of formula (III) (I1.5R+ model). In the
second experiment, animals were divided randomly into two groups,
one group in which the administration started two weeks after the
1.5-hour ischemia/reperfusion and continued for four weeks (I1.5R+2
weeks+dosing for 4 weeks), and the other group for the 1.5-hour
ischemia/reperfusion model without administering the compound of
formula (III) (I1.5R+ model).
[0086] In the third experiment, animals were divided randomly into
two groups, one group in which the administration started three
days after the 1.5-hour ischemia/reperfusion and continued for four
weeks (I1.5R+3 days+dosing for 4 weeks) and the other group for the
1.5-hour ischemia/reperfusion model without administering the
compound of formula (III) (I1.5R+ model). The three treatment
groups with different start times of administration were all dosed
at a dose of 3.6 mg/kg by intraperitoneal injection once a day from
the beginning of administration for four weeks. Monitoring the
scores of neuroethology symptoms 24 hours after the cerebral
ischemia reperfusion injury and at each subsequent week (including
each week during and after the administration) in rats of each
group. The final results and sample size: the combined chart of
five experimental results from the I1.5R+1 week+dosing 4 weeks
group (animal data: I1.5R+1 week group, 31 animals in total); the
chart of experimental result from the I1.5R+2 week+dosing for 4
weeks group (animal data: I1.5R+2 weeks+dosing for 4 weeks group, 9
animals in total).
[0087] The weight and score change after administration of the
compound of formula (III) one week after the 1.5-hour
ischemia/reperfusion for four weeks. The results showed that,
compared to the model group without administering the compound of
formula (III), the animals in the I1.5R+1 week+dosing for 4 weeks
group had a gradually increased weight from the beginning of the
administration and a gradually declined neuroethology score, and
the data results of neuroethology score are statistically
significant in the second, third, fourth, fifth and seventh week
after the operation. The detailed results are shown in FIGS. 3 and
4.
[0088] The weight and score change after administration of the
compound of formula (III) two weeks after the 1.5-hour
ischemia/reperfusion for four weeks. The results showed that,
compared to the model group without administering the compound of
formula (III), the animals in the I1.5R+2 weeks+dosing for 4 weeks
group had a gradually increased weight from the beginning of
administration, however the weight data of the administration group
cannot be demonstrative, since the data of the administration group
had been statistically significant over the model group without
administering the compound of formula (III) before the beginning of
the administration. In the meantime, the neurological scores of the
I1.5R+2 weeks+dosing for 4 weeks group declined from the beginning
of the administration compared to the model group without
administering the compound of formula (III), and the score data are
statistically significant at the fifth week after the operation.
The results are shown in FIGS. 5 and 6. The measurement results of
cerebral infarction injury ratio in rats are shown in FIGS. 7 and
8.
[0089] Based on the above experimental data, it can be seen that
the pharmaceutical composition comprising the compound according to
the present invention has evident effects for the treatment or
improvement of sequelae of ischemic cerebral stroke. Without
departing from the concept of the present invention, suitable
modifications or variations based on the present invention fall
within the scope of the present invention.
* * * * *