CN105998048A - Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof - Google Patents
Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof Download PDFInfo
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- CN105998048A CN105998048A CN201610318781.8A CN201610318781A CN105998048A CN 105998048 A CN105998048 A CN 105998048A CN 201610318781 A CN201610318781 A CN 201610318781A CN 105998048 A CN105998048 A CN 105998048A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7084—Compounds having two nucleosides or nucleotides, e.g. nicotinamide-adenine dinucleotide, flavine-adenine dinucleotide
Abstract
The invention relates to a pharmaceutical composition for treating ischemic cerebral apoplexy and a preparation method and application thereof. The pharmaceutical composition for treating ischemic cerebral apoplexy is prepared from, by weight, 3-12 parts of NADPH and 0.5-8 parts of apocynin. NADPH and apocynin are applied jointly and act cooperatively in a specific proportion, and the pharmaceutical composition can reduce the cerebral infarction volume, significantly relieve the behavior disorder of cerebral ischemia mice, relieve encephaledema, improve the long-term survival capability of mice and enhance neurological function recovery; besides, the treatment effect of the pharmaceutical composition on ischemic cerebral apoplexy is much better than that of NADPH and apocynin which are applied independently on ischemic cerebral apoplexy, and a synergistic effect is achieved when NADPH and apocynin are applied jointly. Thus, the pharmaceutical composition has the effect of treating ischemic cerebral apoplexy and can serve as a potential drug for treating ischemic cerebral apoplexy.
Description
Technical field
The invention belongs to drug world, be specifically related to a kind of pharmaceutical composition treating ischemia apoplexy and
Its preparation method and purposes.
Background technology
Apoplexy (Stroke), also known as apoplexy or cerebrovas-cularaccident, is the brain blood of a kind of unexpected onset
Circulation disorder, is one of three big diseases threatening human health, has sickness rate height, mortality rate
Feature high, that disability rate is high, relapse rate is high.Apoplexy refers to the patient at cerebrovascular disease, because of various
Risk factor causes internal artery narrow, inaccessible or rupture, and causes acute brain disturbance of blood circulation, faces
The sings and symptoms of transient or permanent disordered brain function is shown as on bed.Apoplexy is divided into ischemic brain
Apoplexy and hemorrhagic apoplexy.In recent years, the cerebrovascular patients of China rises year by year, with ischemic
Cerebrovascular is the most common, accounts for 20-25%.At present, China new carbuncle in the occipital region apoplexy 300-350 ten thousand people every year,
Mortality rate is high, and in survivor, about 75% disables, and within 5 years, relapse rate is up to 41%.
At present, China has been enter into senescence society, and the sickness rate of apoplexy will have the trend of increase further.
Therefore, pathomechanism and the treatment protection of research apoplexy especially ischemia apoplexy is always the world of medicine
Vital task.In theory, for medicine (the neural guarantor of cell injury after acute ischemia or Ischemia Reperfusion
Protect agent) brain cell can be protected, improve the toleration to hypoxic-ischemic damage, in terms of current result of study,
Great majority have medicable medicine in zoopery, often end in failure in clinical trial.
Chinese patent literature CN103340890A discloses NADPH as preparation for preventing and treating cerebral ischemia
Application in terms of apoplexy medicine.But, have no that NADPH and 4-hydroxy-3-methoxyacetophenone use in conjunction treatment brain lack
The report of courageous and upright apoplexy.
Therefore, the medicine studying novel treatment cerebral ischemia diseases is significant.
Summary of the invention
To this end, the present invention proposes a kind of pharmaceutical composition treating ischemia apoplexy.
For solving above-mentioned technical problem, the present invention is achieved through the following technical solutions:
The present invention provides a kind of pharmaceutical composition treating ischemia apoplexy, its raw material composition to include:
NADPH 3～12 weight portion, 4-hydroxy-3-methoxyacetophenone 0.5～8 weight portion.
Preferably, the pharmaceutical composition of the present invention above-mentioned treatment ischemia apoplexy, its raw material composition includes:
NADPH 6～9 weight portion, 4-hydroxy-3-methoxyacetophenone 1～4 weight portion.
It is further preferred that the pharmaceutical composition of the present invention above-mentioned treatment ischemia apoplexy, its raw material group
One-tenth includes:
NADPH 7.5 weight portion, 4-hydroxy-3-methoxyacetophenone 2.5 weight portion；Or
NADPH 6 weight portion, 4-hydroxy-3-methoxyacetophenone 4 weight portion；Or
NADPH 9 weight portion, 4-hydroxy-3-methoxyacetophenone 1 weight portion；Or
NADPH 8 weight portion, 4-hydroxy-3-methoxyacetophenone 2 weight portion.
The present invention also provides for the preparation method of the pharmaceutical composition of a kind of above-mentioned treatment ischemia apoplexy, bag
Include following steps:
Taking NADPH and the 4-hydroxy-3-methoxyacetophenone of selected weight portion respectively, mix homogeneously makes mix preparation,
Or be respectively prepared 2 kinds of preparations and use different administering modes.
The present invention also provides for including the preparation of the pharmaceutical composition of above-mentioned treatment ischemia apoplexy or bag
Include the preparation of the pharmaceutical composition that above-mentioned preparation method prepares,
Described pharmaceutical composition adds customary adjuvant, according to common process, makes the most acceptable
Agent, capsule, powder, mixture, pill, granule, solution, syrup, soft extract, plaster
Agent, suppository, aerosol, ointment or injection.
Described pharmaceutically acceptable adjuvant is: filler, disintegrating agent, lubricant, suspending agent, bonding
Agent, sweeting agent, correctives, preservative, substrate etc..Filler includes: starch, pregelatinized Starch,
Lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.；Disintegrating agent includes: starch, pregelatinated
Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl fiber
Element, cross-linked carboxymethyl cellulose are received；Lubricant includes: magnesium stearate, sodium lauryl sulphate, cunning
Stone powder, silicon dioxide etc.；Suspending agent includes: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, fine jade
Fat, hydroxypropyl methyl cellulose etc.；Binding agent includes, starch slurry, polyvinylpyrrolidone, hydroxypropyl
Methylcellulose etc.；Sweeting agent includes: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone
Deng；Correctives includes: sweeting agent and various essence；Preservative includes: parabens, benzoic acid, benzene
Sodium formate, sorbic acid and its esters, benzalkonium bromide, fixed, the eucalyptus oil of acetic acid chloroethene etc.；Substrate includes:
PEG6000, PEG4000, insect wax etc..
The present invention also provides for aforementioned pharmaceutical compositions, above-mentioned preparation method prepares pharmaceutical composition,
Or the application that the preparation of aforementioned pharmaceutical compositions is in the medicine of preparation treatment ischemia apoplexy.
The technique scheme of the present invention has the advantage that compared to existing technology
Though NADPH itself has an effect of certain treatment apoplexy, but NADPH be likely to by
Nadph oxidase utilizes the curative effect producing oxyradical and reduce NADPH treatment apoplexy；Folder
Bamboo Fructus Persicae fiber crops element plays the work for the treatment of apoplexy itself by suppression nadph oxidase reduces ROS generation
With, but its ROS to having produced and the ROS that produced by other approach is (such as mitochondrion and amber
Amber acidohydrogenase) there is no inhibitory action, therefore also there is limitation during treatment apoplexy.
The present invention is by by NADPH and 4-hydroxy-3-methoxyacetophenone administering drug combinations, and the two is total under specific proportioning
Same-action, and act on different target spots, ROS can be removed and can suppress again the generation of ROS.This medicine
Compositions, under the conditions of using the route of administration being suitable for clinical practice, can reduce cerebral infarction volume, aobvious
Work improves cerebral ischemia mice behavior disorder, alleviates cerebral edema, improves ability and the enhancing of mice long term survival
Neurological rehabilitation；And, this pharmaceutical composition is significantly better than two to the therapeutic effect of ischemia apoplexy
The individually dosed therapeutic effect to ischemia apoplexy of person, NADPH and 4-hydroxy-3-methoxyacetophenone administering drug combinations tool
There is the effect of Synergistic.This shows, this pharmaceutical composition has the effect for the treatment of ischemia apoplexy,
Can be as the medicine of potential treatment ischemia apoplexy.
Accompanying drawing explanation
In order to make present disclosure be more likely to be clearly understood, being embodied as below according to the present invention
Example also combines accompanying drawing, and the present invention is further detailed explanation, wherein:
Fig. 1 (a), 1 (b) are NADPH associating Apocynin intracerebroventricular injection and intravenous administration
Impact on ischemia apoplexy mouse brain Infarction volume, wherein, * represents that p < 0.05, * * represents
P < 0.01, * * * represents that p < 0.001, ## represents that p < 0.01, I.C.V represents that intracerebroventricular injection is administered, I.V
Represent intravenous administration；
Fig. 2 (a), 2 (b) are that NADPH associating Apocynin intravenous administration is in ischemic brain
The impact of wind mouse brain Infarction volume, wherein, * represents that p < 0.05, * * represents that p < 0.01, * * * represents
P < 0.001, △ represents p < 0.05；
Fig. 3 is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy mouse Nerve disease
The impact of shape, wherein, * represents that p < 0.05, * * represents that p < 0.01, * * * represents that p < 0.001, △ represents
p<0.05；
Fig. 4 is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy mouse brain edema
Impact, wherein, * represents that p < 0.05, * * * represents that p < 0.001, ### represents p < 0.001；
Fig. 5 is that ischemia apoplexy mice is deposited by NADPH associating Apocynin intravenous administration for a long time
The impact lived, wherein, * represents that p < 0.05, # represents p < 0.05；
Fig. 6 is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy mice balance fortune
Dynamic impact, wherein, * represents that p < 0.05, * * * represents that p < 0.001, ### represents p < 0.001, △ table
Show p < 0.05；
Fig. 7 is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy mice study note
Recalling the impact of ability, wherein, * * represents that p < 0.01, * * * represents that p < 0.001, ### represents p < 0.001,
△ represents that p < 0.05, △ △ represents p < 0.01；
Fig. 8 is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy mouse brain skin
The impact of layer ischemic region ROS level, wherein, * * * represents that p < 0.001, ### represents p < 0.001, △
Represent p < 0.05；
Fig. 9 (a), 9 (b), 9 (c), 9 (d), 9 (e), 9 (f) are NADPH associatings
Apocynin intravenous administration is to ischemia apoplexy Cerebral Cortex ischemic region NOX2, NOX4
The impact of protein level, wherein, * represents that p < 0.05, * * represents that p < 0.01, * * * represents p < 0.001,
## represents that p < 0.01, ### represents that p < 0.001, △ represents that p < 0.05, △ △ represents p < 0.01；
Figure 10 (a), 10 (b), 10 (c), 10 (d), 10 (e), 10 (f), 10 (g), 10
(h)、10(i)、10(j)、10(k)、10(l)、10(m)、10(n)、10(o)、10(p)
It is that NADPH associating Apocynin intravenous administration is to ischemia apoplexy Cerebral Cortex ischemic region
The impact of NALP3 inflammation complex pathway protein level, wherein, * represents that p < 0.05, * * represents
P < 0.01, # represents that p < 0.05, ## represents that p < 0.01, ### represents that p < 0.001, △ represents p < 0.05,
△ △ represents p < 0.01；
In Fig. 1-Figure 10, what Apocynin represented is: 4-hydroxy-3-methoxyacetophenone.
Detailed description of the invention
Embodiment 1
The pharmaceutical composition of the present embodiment treatment ischemia apoplexy, its raw material consists of: NADPH
7.5g, 4-hydroxy-3-methoxyacetophenone 2.5g；
Its preparation method, comprises the following steps: take NADPH and the apocynum cannabinum of selected weight portion respectively
Element, mix homogeneously, to obtain final product.
Embodiment 2
The pharmaceutical composition of the present embodiment treatment ischemia apoplexy, its raw material consists of: NADPH 6g,
4-hydroxy-3-methoxyacetophenone 4g；
Its preparation method, comprises the following steps: take NADPH and the apocynum cannabinum of selected weight portion respectively
Element, mix homogeneously, to obtain final product.
Embodiment 3
The pharmaceutical composition of the present embodiment treatment ischemia apoplexy, its raw material consists of: NADPH 9g,
4-hydroxy-3-methoxyacetophenone 1g；
Its preparation method, comprises the following steps: take NADPH and the apocynum cannabinum of selected weight portion respectively
Element, mix homogeneously, to obtain final product.
Embodiment 4
The pharmaceutical composition of the present embodiment treatment ischemia apoplexy, its raw material consists of: NADPH 8g,
4-hydroxy-3-methoxyacetophenone 2g；
Its preparation method, comprises the following steps: take NADPH and the apocynum cannabinum of selected weight portion respectively
Element, mix homogeneously, to obtain final product.
Comparative example 1
The pharmaceutical composition of the present embodiment treatment ischemia apoplexy, its raw material consists of: NADPH 1g,
4-hydroxy-3-methoxyacetophenone 4g；
Its preparation method, comprises the following steps: take NADPH and the apocynum cannabinum of selected weight portion respectively
Element, mix homogeneously, to obtain final product.
Experimental example
Following each experimental example proves technique effect of the present invention.
In the following experimental example of the present invention, what Apocynin represented is: 4-hydroxy-3-methoxyacetophenone.
Experimental example 1The protective effect that Cerebral Ischemia-reperfusion in Mice is damaged by NADPH associating 4-hydroxy-3-methoxyacetophenone
(1) experiment material
Cleaning grade male ICR mouse, quality 23～28g, University Of Suzhou's Experimental Animal Center provides, real
Test animal productiong credit number: XCYK (Soviet Union) 2002-2008, laboratory animal use credit number:
SYXK (Soviet Union) 2002-0037.
Room temperature 22 DEG C, humidity 50-60%, well-ventilated, the most round the clock (12h/12h), freely ingest and take the photograph
Water.Before experiment, hero mice is adapted in feeding environment 2d.
Divide 6 groups: sham operated rats, model group, NADPH group (7.5mg/kg), 4-hydroxy-3-methoxyacetophenone group
(2.5mg/kg), NADPH+ 4-hydroxy-3-methoxyacetophenone group (7.5mg+2.5mg/kg), NADPH+ apocynum cannabinum
Element group (1mg+4mg/kg).
The source of exogenous NADPH and 4-hydroxy-3-methoxyacetophenone medicine can pass through synthetic, semi-synthetic,
Biological extraction obtains.
(2) experimental program
1) mice transience middle cerebral artery occlusion model is set up
Using internal carotid artery line brush, mouse MCAO model is prepared in improvement a little, and mice is with 4% hydration
Chloral (400mg/kg) intraperitoneal injection of anesthesia, we use line brush to prepare ischemia model, separate neck total,
Neck is outer and internal carotid artery, and ligation strength is outer and the total proximal part of neck, line bolt (6023, Doccol Corporation,
Redlands, USA) insert from neck until anterior cerebral artery initiating terminal, block middle cerebral artery blood supply.
After blocking blood flow 2h, pull out Outlet bolt and realize Reperfu-sion.Sham operated rats mice in addition to not plug wire, remaining step
Rapid all identical with ischemia group and treatment group.In whole operation process, room temperature is maintained at 22～25 DEG C, uses certainly
Move temperature control heating cushion by mice anus temperature control at 37 ± 0.5 DEG C.Postoperative being placed in by animal is placed with cleaning bedding and padding
Raise in box, freely drink water, take food.
2) cerebral infarction volume measures
After cerebral ischemia re-pouring 24h by mice broken end take brain, put refrigerator (-20 DEG C) several minutes, remove olfactory bulb,
Cerebellum and low brain stem, crown 4 cuttves of cutting are divided into 5 (2mm), and brain sheet red tetrazolium (TTC) contaminates
Color, dyeing liquor consists of: 1.5mL1%TTC, 0.1mL1mol/L K2HPO4, 3.4mL normal saline,
37 DEG C of lucifuge dyeing 30min, normal structure takes on a red color, and infarction tissue is white.
4% formaldehyde is fixing two days later, blots liquid with filter paper and further takes out cerebral infarction tissue, with infarcted cerebral constitution
Weight accounts for the percentage ratio index as cerebral infarction volume of total cerebral weight.With Sigma Pro5.0 software meter
Calculate cerebral infarct size percentage.
3) nervous symptoms scoring
After Cerebral Ischemia-reperfusion in Mice 24h, an observer not knowing about packet situation comment according to the five-grade marking system
Minute mark is accurate carries out neurobehavioral obstacle scoring to mice: 0 point: impassivity functional impairment symptom；1 point: no
Can full extension offside forelimb；2 points: rotate to offside during walking, " knocking into the back " phenomenon occurs；3 points:
Astasia, topples over to offside；4 points: can not spontaneous walk, disturbance of consciousness.
4) brain moisture determination
After cerebral ischemia re-pouring 24h, mice broken end is taken brain, remove olfactory bulb, cerebellum and low brain stem, claim
Take brain weight in wet base, after 107 DEG C of baking 48h, claim dry weight, brain PCm=(weight in wet base-dry weight)/
Weight in wet base × 100%.
5) function of nervous system's test
I, the making of animal model
Choose the body weight male ICR mouse at 23-28g, make the operation of cerebral ischemia 2h Reperfu-sion.This animal
Model ischemic time is long, and mortality rate is high, notes supplying in time number of animals.
II, motor function are tested
Rotor shaft test (Rota-rod test) keeps balance to need proprioceptive sensation, topognosis in rotor shaft
And fine setting motor capacity.This test request mice keep at the uniform velocity swingle balance, and record its turn
Movement time and drop-out time on rod obtain transfer rod rotary speed.Test is accelerated slowly limit individuality
Between performance diversity.
Required instrument includes with material: (1) instrument: a diameter of about 5cm in roller axle center, by heavily fortified point
Solid plastic production form, outside parcel Lycoperdon polymorphum Vitt rubbery foam, Guan Kuanyue 5cm, this instrument is permissible
40 turns/min is accelerated to from 4 turns/min in 300s；(2) stopwatch；(3) 50% ethanol；(4)
Napkin.
Test when, mice is placed in themselves cage, makes them suitable in test room
Answer 15min (adaptation environmental phase).Whole test is made up of three tests of interval 15min.?
The training stage is not had before test session.Next group mice directly can be grasped by it when same single test
Make.Instrument is set as in 300s accelerating to 40 turns/min from 4 turns/min.Instrument is before start with 4
Turn/the constant speed of min operating.
III, the Behavior Test of learning and memory practise maze experiment (Y-maze test)
The end of Y type electric maze 3 arm all has l lamp, and bottom is electrical network, the most only 1 arm end
Lamp send light, now bottom this arm, electrical network no current passes through, and is place of safety；The lamp of another two-arm is not
Bright, bottom electrical network energising (about 50V), for non-security district；Place of safety changes at random with non-security district.
When experiment starts, animal is put into labyrinth adapts in arbitrary arm 2～3min；Then other are arbitrary
The signal lights of arm is opened as conditional stimulus, and after ls postpones, (unconditional is stung in the two-arm energising that lamp does not works
Swash)；Shocking by electricity to place of safety when animal escapes, lamp is bright continues 15s, and then light-off rest 45s, the completeest
Become 1 operation and record the time used；Start next operation the most again.
6) data statistics and analysis
Data all represent with mean ± SD (Mean ± SD), and statistical analysis uses one factor analysis of variance
(one-way ANOVA), p < 0.05 has significance for significant difference.
(4) experimental result NADPH associating 4-hydroxy-3-methoxyacetophenone intracerebroventricular injection and intravenous administration are to lacking
Shown in impact such as Fig. 1 (a), 1 (b) of courageous and upright apoplexy mouse brain Infarction volume.
From Fig. 1 (a), 1 (b), compared with model group, intracerebroventricular injection NADPH
(1mg/kg) associating 4-hydroxy-3-methoxyacetophenone (4mg/kg) medication group significantly reduces Cerebral Ischemia-reperfusion in Mice 24
H cerebral infarction volume (p < 0.05)；4-hydroxy-3-methoxyacetophenone is combined with intravenous injection NADPH (1mg/kg)
(4mg/kg) the NADPH associating the brightest focal cerebral ischemia that significantly reduces of 4-hydroxy-3-methoxyacetophenone medication group fills again
Note 24h cerebral infarction volume (p < 0.05, p < 0.01)；Intravenous injection NADPH (7.5mg/kg) combines
4-hydroxy-3-methoxyacetophenone (2.5mg/kg) medication group has been significantly reduced cerebral infarction after Cerebral Ischemia-reperfusion in Mice 24h
Dead volume (p < 0.001).But it is compared to each other the result of three groups it is found that intravenous injection NADPH
(1mg/kg) effect that associating 4-hydroxy-3-methoxyacetophenone (4mg/kg) produces is than the difference of intracerebroventricular, and difference has
Significance (p < 0.05)；Intravenous injection NADPH (7.5mg/kg) combines 4-hydroxy-3-methoxyacetophenone
(2.5mg/kg) effect produced combines 4-hydroxy-3-methoxyacetophenone than intravenous injection NADPH (1mg/kg)
(4mg/kg) produce is effective, and difference has significance (p < 0.01).Therefore, independent intravenous injection
NADPH (1mg/kg) does not significantly reduce the effect of cerebral infarction volume, independent intravenous injection Folium seu Cortex Nerii
Fiber crops element (2.5mg/kg) only have slight therapeutic effect, so intravenous injection NADPH (7.5mg/kg)
Associating 4-hydroxy-3-methoxyacetophenone (2.5mg/kg) effect more has clinical value.
4-hydroxy-3-methoxyacetophenone 4-hydroxy-3-methoxyacetophenone 4-hydroxy-3-methoxyacetophenone NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous injection is given
Medicine is shown on impact such as Fig. 2 (a), 2 (b) of ischemia apoplexy mouse brain Infarction volume.
From Fig. 2 (a), 2 (b), compared with model group, NADPH group and 4-hydroxy-3-methoxyacetophenone group
All significantly reduce Cerebral Ischemia-reperfusion in Mice 24h cerebral infarction volume (p < 0.01)；NADPH combines folder
Bamboo Fructus Persicae fiber crops element group reduces mouse brain Infarction volume (p < 0.001) further；Compared with independent medication group, associating
Medication group has significant difference (p < 0.05).This shows, NADPH, 4-hydroxy-3-methoxyacetophenone and both connection
Share medicine and all can reduce cerebral infarction mouse brain Infarction volume, the effect that two medicines share is better than individually making
With.
NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration is to ischemia apoplexy mouse Nerve symptom
Impact is as shown in Figure 3.
From the figure 3, it may be seen that compared with model group, NADPH group and 4-hydroxy-3-methoxyacetophenone group substantially reduce mice
After cerebral ischemia re-pouring 24h, nervous symptoms comments (p < 0.05)；Both then reduce further little at use in conjunction
Mus nervous symptoms scoring (p < 0.01)；Compared with independent medication group, drug combination group has significant difference
(p<0.05).This shows, NADPH, 4-hydroxy-3-methoxyacetophenone and both use in conjunction all can be improved brain and lack
The nervous symptoms of courageous and upright apoplexy mice, the effect that two medicines share is better than being used alone.
NADPH combines the 4-hydroxy-3-methoxyacetophenone intravenous administration shadow to ischemia apoplexy mouse brain edema
Ring as shown in Figure 4.
As shown in Figure 4, compared with model group, NADPH group and 4-hydroxy-3-methoxyacetophenone group significantly reduce little
Big brain water content (p < 0.05) after Mus cerebral ischemia re-pouring 24h；Both reduce the most further use in conjunction
The big brain water content of mice (p < 0.01).This shows, NADPH, 4-hydroxy-3-methoxyacetophenone and both combine should
With all reducing the cerebral edema of cerebral infarction mice, the effect that two medicines share is better than being used alone.
NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration is to ischemia apoplexy mice long term survival
Impact is as shown in Figure 5.
As shown in Figure 5, compared with model group, NADPH group and 4-hydroxy-3-methoxyacetophenone group and two medicines are share
Group significantly improves 28 days survival rates (p < 0.05) after Cerebral Ischemia-reperfusion in Mice.This shows, NADPH,
4-hydroxy-3-methoxyacetophenone and both use in conjunction all can improve mouse survival rate, and the effect that two medicines share is better than list
Solely use.
NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration is statokinetic to ischemia apoplexy mice
Impact is as shown in Figure 6.
It will be appreciated from fig. 6 that compared with model group, NADPH, 4-hydroxy-3-methoxyacetophenone group clearly enhance cerebral ischemia
The balance exercise ability (p < 0.05) of 28 days survival mice after Reperfu-sion, NADPH combines 4-hydroxy-3-methoxyacetophenone
Group more significantly enhances the balance exercise ability (p < 0.001) of 28 days survival mice after cerebral ischemia re-pouring；
Compared with independent medication group, drug combination group has significant difference (p < 0.05).This shows, NADPH,
4-hydroxy-3-methoxyacetophenone and both share and all can improve mice balance exercise ability, the effect that two medicines share is better than
It is used alone.
NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration is to ischemia apoplexy learning and memory of little mouse energy
The impact of power is as shown in Figure 7.
As shown in Figure 7, compared with model group, NADPH, 4-hydroxy-3-methoxyacetophenone group clearly enhances cerebral ischemia
The ability of learning and memory (p < 0.01) of 28 days survival mice after Reperfu-sion, NADPH combines 4-hydroxy-3-methoxyacetophenone
Group more significantly improves the ability of learning and memory (p < 0.001) of 28 days survival mice after cerebral ischemia re-pouring；
Compared with independent medication group, drug combination group has significant difference (p < 0.05).This shows, NADPH
Associating 4-hydroxy-3-methoxyacetophenone can more effectively improve ability of learning and memory in mice.
Experimental example 2NADPH associating 4-hydroxy-3-methoxyacetophenone is to ischemia apoplexy Cerebral Cortex ischemic region
The impact of ROS level
(1) experiment material
With experimental example 1.
(2) experimental program
1) mice transience middle cerebral artery occlusion model is set up
With experimental example 1.
2) mensuration of ROS
3h after mice Ischemia Reperfusion, after deep anaesthesia, broken end takes brain, removes olfactory bulb, cerebellum and low brain stem,
Freezing microtome is cut into slices, and thick 10 μm pasters, on the microscope slide that gelatin processed, then use frozen section
Oxidative stress active oxygen fluorimetric reagent box detects, i.e. in fluorescence microscopy Microscopic observation (qualitative detection):
Excitation wavelength 490nm, distributes wavelength 530nm, Fluorescence Increasing, shows activity chalcogen (ROS) content
High.
(3) experimental result
Ischemia apoplexy Cerebral Cortex is lacked by NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration
The impact of blood district ROS level is as shown in Figure 8.
As shown in Figure 8, compared with sham-operation (sham) group, after Model group Cerebral Ischemia-reperfusion in Mice 3h
Significantly improve ROS level (p < 0.001)；Application NADPH, after 4-hydroxy-3-methoxyacetophenone, with Model group
Compare, all significantly reduce the rising (p < 0.001) of ischemia apoplexy Cerebral Cortex ROS level,
Use in conjunction NADPH and 4-hydroxy-3-methoxyacetophenone significantly more inhibit ischemia apoplexy Cerebral Cortex
The rising (p < 0.001) of ROS level.Result prompting NADPH, 4-hydroxy-3-methoxyacetophenone, and both combine use
Medicine, all can reduce ischemia apoplexy Cerebral Cortex ischemic region ROS level, the effect that two medicines share
It is better than being used alone.
Experimental example 3NADPH associating 4-hydroxy-3-methoxyacetophenone is to ischemia apoplexy Cerebral Cortex ischemic region
The impact of NOx level
(1) experiment material
With experimental example 1.
(2) experimental program
1) mice transience middle cerebral artery occlusion model is set up
With experimental example 1.
2) Western blot method
A. 12%SDS-PAGE, loading, voltage spacer gel 90V, separation gel 110V, electrophoresis are prepared；
B. use wet robin that sample is transferred to NC film after electrophoresis terminates, the glue constant current 0.5A of 10%, 1.5
h；The glue constant current 0.3A, 45min of 14%；
C., after transferring film terminates, NC film is put into the TBS containing 5% defatted milk powder closes under room temperature 2h；
D. adding one of the TBS configuration containing 5% defatted milk powder and 0.1% sodium azide to resist, 4 DEG C overnight.
E.TBS-T rinses 15min × 3 time, and TBS rinses 15min × 1 time；
F. add and resist with containing the two of 5% defatted milk powder TBS configuration, room temperature 2h；
G.TBS-T rinses 15min × 3 time, and TBS rinses 15min × 1 time, and film is placed in ECL colour developing
In liquid (before use A, B liquid equal-volume mixing), room temperature 1min, tabletting, expose, develop.
(3) experimental result
Ischemia apoplexy Cerebral Cortex is lacked by NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration
The impact of blood district NOX2, NOX4 protein level such as Fig. 9 (a), 9 (b), 9 (c), 9 (d), 9
Shown in (e), 9 (f).
From Fig. 9 (a), 9 (b), 9 (c), 9 (d), 9 (e), 9 (f), sham operated rats
The expression of NOX2 and NOX4 is relatively low, and ischemia-reperfusion 8h, 16h, NOX2 protein expression substantially rises
High (p < 0.01), and NOX4 protein expression is significantly to raise (p < 0.001) at 16h.Give NADPH,
4-hydroxy-3-methoxyacetophenone, and administering drug combinations, compared with Model group, NADPH, 4-hydroxy-3-methoxyacetophenone and associating
It is administered the expression inhibiting NOX2 and NOX4 the most in various degree.This shows, NADPH, presss from both sides bamboo
Fructus Persicae fiber crops element, and both drug combinations, all can the up-regulated of restraint NO X 2 and NOX4, two medicines close
Effect be better than being used alone.
Experimental example 4NADPH associating 4-hydroxy-3-methoxyacetophenone is to ischemia apoplexy Cerebral Cortex ischemic region
NALP3 inflammation complex associated component and the impact of inflammatory factor protein level
(1) experiment material
With experimental example 1.
(2) experimental program
1) mice transience middle cerebral artery occlusion model is set up
With experimental example 1.
2) Western blot method
With experimental example 2.
(3) experimental result
Ischemia apoplexy Cerebral Cortex is lacked by NADPH associating 4-hydroxy-3-methoxyacetophenone intravenous administration
Impact such as Figure 10 (a), 10 (b) of blood district NALP3 inflammation complex pathway protein level, 10 (c),
10(d)、10(e)、10(f)、10(g)、10(h)、10(i)、10(j)、10(k)、10
Shown in (l), 10 (m), 10 (n), 10 (o), 10 (p).
By Figure 10 (a), 10 (b), 10 (c), 10 (d), 10 (e), 10 (f), 10 (g), 10
(h)、10(i)、10(j)、10(k)、10(l)、10(m)、10(n)、10(o)、10(p)
Understanding, sham operated rats NLRP3 protein expression is relatively low, and in the cerebral tissue of I/R group mice ischemia side
NLRP3, ASC, precursor caspase-1, and the protein level of il-1b, il-18 has at 8h and 16h
Dramatically increase.These results suggest that, ischemia induction the little body protein of inflammation, IL-1 increase, IL-18 inflammation because of
The expression of son increases.Give NADPH, 4-hydroxy-3-methoxyacetophenone, and administering drug combinations, with model group phase
Ratio, NADPH, 4-hydroxy-3-methoxyacetophenone and administering drug combinations in various degree reduce NLRP3, ASC,
Precursor caspase-1 cracks caspase-1 and il-1b, the protein level of il-18.This shows, NADPH,
4-hydroxy-3-methoxyacetophenone, and administering drug combinations all can suppress the activation of NLRP3 inflammation complex path and inflammation because of
The generation of son, the effect that two medicines share is better than being used alone.
To sum up, the present invention is by by NADPH and 4-hydroxy-3-methoxyacetophenone administering drug combinations, and the two is specifically being joined
The lower common effect of ratio, this pharmaceutical composition can reduce cerebral infarction volume, significantly improves cerebral ischemia mice behavior
Obstacle, alleviates cerebral edema, improves the ability of mice long term survival and strengthens neurological rehabilitation；And,
It is individually dosed to ischemic brain that this pharmaceutical composition is significantly better than the two to the therapeutic effect of ischemia apoplexy
The therapeutic effect of apoplexy, NADPH and 4-hydroxy-3-methoxyacetophenone administering drug combinations have the effect of Synergistic.This
Showing, this pharmaceutical composition has the effect for the treatment of ischemia apoplexy, can lack as potential treatment
The medicine of courageous and upright apoplexy.
Obviously, above-described embodiment is only for clearly demonstrating example, and not to embodiment
Restriction.For those of ordinary skill in the field, can also do on the basis of the above description
Go out change or the variation of other multi-form.Here without also cannot all of embodiment be given exhaustive.
And the obvious change thus extended out or variation still in the protection domain of the invention among.
Claims (6)
1. a pharmaceutical composition, it is characterised in that its raw material composition includes:
NADPH 3～12 weight portion, 4-hydroxy-3-methoxyacetophenone 0.5～8 weight portion.
Pharmaceutical composition the most according to claim 1, it is characterised in that its raw material composition includes:
NADPH 6～9 weight portion, 4-hydroxy-3-methoxyacetophenone 1～4 weight portion.
Pharmaceutical composition the most according to claim 2, it is characterised in that its raw material composition includes:
NADPH 7.5 weight portion, 4-hydroxy-3-methoxyacetophenone 2.5 weight portion；Or
NADPH 6 weight portion, 4-hydroxy-3-methoxyacetophenone 4 weight portion；Or
NADPH 9 weight portion, 4-hydroxy-3-methoxyacetophenone 1 weight portion；Or
NADPH 8 weight portion, 4-hydroxy-3-methoxyacetophenone 2 weight portion.
4. the preparation method of the pharmaceutical composition described in an any one of claim 1-3, it is characterised in that
Comprise the following steps:
Take NADPH and the 4-hydroxy-3-methoxyacetophenone of selected weight portion, mix homogeneously respectively, to obtain final product.
5. include the preparation of the pharmaceutical composition described in any one of claim 1-3 or include that right is wanted
Seek the preparation of the pharmaceutical composition that the preparation method described in 4 prepares, it is characterised in that
Described pharmaceutical composition adds customary adjuvant, according to common process, makes the most acceptable
Agent, capsule, powder, mixture, pill, granule, solution, syrup, soft extract, plaster
Agent, suppository, aerosol, ointment or injection.
6. the pharmaceutical composition described in any one of claim 1-3, the preparation method described in claim 4
The preparation of the pharmaceutical composition described in the pharmaceutical composition prepared or claim 5 is in preparation treatment
Application in the medicine of ischemia apoplexy.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610318781.8A CN105998048A (en) | 2016-05-13 | 2016-05-13 | Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof |
| PCT/CN2016/109005 WO2017193573A1 (en) | 2016-05-13 | 2016-12-08 | Pharmaceutical composition for treating ischemic stroke, and preparation method and application thereof |
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| CN201610318781.8A CN105998048A (en) | 2016-05-13 | 2016-05-13 | Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017193573A1 (en) * | 2016-05-13 | 2017-11-16 | 重庆纳德福实业集团股份有限公司 | Pharmaceutical composition for treating ischemic stroke, and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1774243A (en) * | 2003-02-28 | 2006-05-17 | 霍华德弗洛里实验生理及医药学院 | Therapeutic compositions |
| CN103340890A (en) * | 2013-06-08 | 2013-10-09 | 苏州人本药业有限公司 | Application of NADPH in preparing medicaments for preventing and treating ischemic cerebral stroke |
| CN104840478A (en) * | 2015-02-17 | 2015-08-19 | 苏州人本药业有限公司 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
| CN105520938A (en) * | 2006-11-13 | 2016-04-27 | Pcb联合公司 | Composition for inhibiting nadph oxidase activity |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0603975D0 (en) * | 2006-03-01 | 2006-04-05 | Etren | Methods and agents for reducing oxidative stress |
| CN102459291B (en) * | 2009-04-27 | 2015-11-25 | Mcw研究基金会股份有限公司 | Neuroprotective compound and their application |
| CN105998048A (en) * | 2016-05-13 | 2016-10-12 | 重庆纳德福实业集团股份有限公司 | Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof |
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- 2016-12-08 WO PCT/CN2016/109005 patent/WO2017193573A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1774243A (en) * | 2003-02-28 | 2006-05-17 | 霍华德弗洛里实验生理及医药学院 | Therapeutic compositions |
| CN105520938A (en) * | 2006-11-13 | 2016-04-27 | Pcb联合公司 | Composition for inhibiting nadph oxidase activity |
| CN103340890A (en) * | 2013-06-08 | 2013-10-09 | 苏州人本药业有限公司 | Application of NADPH in preparing medicaments for preventing and treating ischemic cerebral stroke |
| CN104840478A (en) * | 2015-02-17 | 2015-08-19 | 苏州人本药业有限公司 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
Non-Patent Citations (1)
| Title |
|---|
| 刘艳丽等: "夹竹桃麻素预处理对急性脑梗死脑组织损伤影响的初步研究", 《中国现代药物应用》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017193573A1 (en) * | 2016-05-13 | 2017-11-16 | 重庆纳德福实业集团股份有限公司 | Pharmaceutical composition for treating ischemic stroke, and preparation method and application thereof |
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Address after: No. 1-2, No. 25, Jinjian Road, Wanshengjing Kai District, Chongqing City, 400800 Applicant after: Chongqing Nadford Industrial Group Co., Ltd. Address before: 400800 Qijiang District, Chongqing (formerly Wansheng District) No. 1-2, No. 25, Qinjian Road, Wanshengjing Kai District Applicant before: Chongqing Telford industrial group Limited by Share Ltd |
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