CN106580956A - Application of Sola A in ischemic brain damage - Google Patents

Abstract

The invention discloses an application of Sola A in ischemic brain damage. Specifically, the invention discloses (I) the usage of the Sola A in producing a pharmaceutical composition used to prevent and/or treat ischemic brain damage and/or (II) the function of the Sola A in preparing a pharmaceutical composition used to protect ischemic neuron. According to experiments, Sola A can effectively control the development of acute brain death, reduce infarct size and slow down neuronal edema and angioedema. The Sola A also plays an positive part in protecting neurological functions for a long time.

Description

Applications of the Suo Lafen A in ischemic brain damage

Technical field

The present invention relates to field of pharmaceutical biology.In particular it relates to antineoplastic Suo Lafen A are to ischemic brain damage In new application.

Background technology

Nearest national coroner's inquest shows, ischemic brain damage (cerebral ischemic injury) or ischemic Cerebral apoplexy (ischemic stroke) has leapt to disables and first of the cause of death for China human mortality, and morbidity has and increases year by year Many trend.Acute ischemic cerebral apoplexy is modal stroke types, accounts for the 60%～80% of whole cerebral apoplexies.But mesh The front medicine to acute apoplexy with definite therapeutic effect only only has rt-PA (recombinant tissue-plasminogen activator, rt-PA) is a kind of.Rt-PA be also current U.S.'s food with Unique medicine of drug administration (US FDA) approval treatment acute apoplexy.But the therapeutic time window of rt-PA is extremely limited. In order to avoid the Complications of Cerebral Hemorrhage that rt-PA therapy-relateds occur, it is presently recommended that it is after acute apoplexy morbidity in 4.5 hours Using.Thus only about less than 3% acute stroke patients just have an opportunity to be treated using rt-PA.

Therefore, research and develop new safely and effectively Treatment of Cerebral Stroke strategy and seem urgent all the more.

The content of the invention

The invention provides a kind of new application of antineoplastic Suo Lafen-A, i.e., it is used to mitigate cerebral arterial thrombosis damage Therapeutic strategy.

A kind of first aspect present invention, there is provided the use of Suo Lafen-A (Soraphen-A) or its pharmaceutically acceptable salt On the way, for prepare (i) prevention and/or treat ischemic brain damage pharmaceutical composition；And/or (ii) protection ischaemic neuronal Pharmaceutical composition, wherein, the Suo Lafen-A have structure shown in Formulas I：

In another preference, described ischemic brain damage includes that Acute ischemia rePerfusion or chronic ischemic brain are damaged Wound.

In another preference, the Acute ischemia rePerfusion includes the ischemic that disease time is between -3 days 2 hours Property brain damage.

In another preference, the chronic ischemic brain damage includes 3 days ischemic brain damages afterwards of disease time.

In another preference, described ischemic brain damage includes cerebral apoplexy, ischemia reperfusion injury, transience brain Ischemic episode, neonatal hypoxic ischemic encephalopathy.

In another preference, the prevention and/or treatment ischemic brain damage include one or more of to ischemic Property brain damage performance improvement：

A () reduces Brain stem injury, such as reduce Infarction volume and/or area；

B () improves limbs symmetry；

C () improves limbs and uses Preference；

D () improves neural sensation and motor function is damaged；

E () increases infarct location brain blood flow；

F () improves and remember after brain damage；

G () reduces the loss of neuron after cerebral ischemia.

In another preference, the application dosage of the Suo Lafen-A is 1-100mg/kg/ days.

In another preference, the application dosage of the Suo Lafen-A is preferably, daily 1-50mg/kg, more preferably for 1-5mg/kg。

In another preference, described pharmaceutical composition contains Suo Lafen-A or its pharmaceutically acceptable salt as work Property composition, and pharmaceutically acceptable carrier.

In another preference, described pharmaceutical composition also contains thrombolytics, rtPA and/or neuroprotective agent.

In another preference, cerebral apoplexy Advance of Intravascular Stent in Treating medicine box is also contained in described pharmaceutical composition.

Second aspect present invention, there is provided a kind of method that external non-therapeutic protects ischaemic neuronal, including step： Ischaemic neuronal is cultivated under conditions of containing Suo Lafen-A or its pharmaceutically acceptable salt, and/or at Jing ischemics Suo Lafen-A are added in the neuronal cultures of reason, so as to protect ischaemic neuronal.

In another preference, described protection includes that preventive protection and/or therapeutic are protected.

Third aspect present invention, there is provided a kind of (i) prevention and/or treatment ischemic brain damage；And/or protection ischemic The method of neuron, including step, apply the Suo Lafen-A of safe and effective amount or its are pharmaceutically acceptable to required object Salt, or containing Suo Lafen-A or its pharmaceutically acceptable salt as active component pharmaceutical composition.

Such as in another preference, the required object includes the mammal with ischemic brain damage, little Mouse, rat or people.

It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and have in below (eg embodiment) Can be combined with each other between each technical characteristic of body description, so as to constitute new or preferred technical scheme.As space is limited, exist This no longer tires out one by one states.

Description of the drawings

Fig. 1 shows action sites of the Suo Lafen-A in cellular process.

Fig. 2 shows the brain blood flow situation of change monitored using laser speckle Doppler technology in MCAO surgical procedures.Its In, Fig. 2A-B show that Bilateral hemispheres brain blood flow is without significant changes after Sham group mouse row sham-operations；And in DMSO groups and In SorA group mouse, the obstruction of arteria cerebri media causes the significant brain blood flow of side cerebral hemisphere and reduces, and brain blood flow Reduction degree is in this two groups of mouse and there was no significant difference；Fig. 2 C-D show that the cerebral infarction volume of Jing SorA treatment mouse is obvious Less than DMSO control mices.

Fig. 3 shows that the ratio that the cerebral ischemia mouse that Sor-A is treated turns to the left is less than the ratio of DMSO groups, represents The limbs symmetry for the treatment of mouse is better than control mice.

Fig. 4 A-B show that compared with DMSO mouse the cerebral ischemia mouse offside forelimb and hind leg of Jing Sor-A treatments are wrong to be walked Number accounts for the percentage of total mistake step number and significantly reduces, and is close to sham groups mouse.

Fig. 5 shows cerebral ischemia mouse after injury, and non-damaging side (right side) limbs preference use ratio is significantly raised, The preference use ratio of Sor-A treatment mouse limbs is substantially less than DMSO control groups.

Fig. 6 A are limbs proprioception function score, and Fig. 6 B are climbing test scoring, and Fig. 6 C are forelimb walking experiment scoring, Fig. 6 D are that side turns experiment scoring, and Fig. 6 E are the comprehensive grading of mouse, and Fig. 6 F are sticky paper experiment scoring；The above results show：Sor-A Treatment is remarkably improved nervous function comprehensive grading after the chronic ischemic brain damage of mouse.

Fig. 7 A-B show Morris water maze tests, as a result show, Sor-A significantly improves mouse in target quadrant The time of staying, therefore, it is possible to significantly improve learning and memory function；Fig. 7 C show the swimming rate of three groups of mouse without notable Sex differernce.

Fig. 8 A-B show that the neuron of survival in the ischemic area of the mouse that application of Suo Lafen-A in advance is significantly more than DMSO control group mices.

Specific embodiment

The present inventor passes through extensively and in-depth study, first it was unexpectedly observed that antineoplastic Suo Lafen-A are for scarce Brain after courageous and upright brain damage has good neuroprotection.It is demonstrated experimentally that to ischemic brain damage acute stage, chronic phase And ischemical reperfusion injury has preferable neuroprotection, occur using Suo Lafen-A or in ischemic brain damage in advance Suo Lafen-A are used afterwards, can effectively be reduced ischemic focus area, be mitigated post-ischemic vascular oedema, and to long-lasting nerve function Repair and there is the good effect being highly profitable.On this basis, the present invention is completed.

Suo Lafen A or its pharmaceutically acceptable salt

As used herein, term " inventive compound ", " Sora-A ", " Suo Lafen A " are used interchangeably, and refer to tool There are the compound or its pharmaceutically acceptable salt of structure shown in Formulas I.

Suo Lafen-A (Soraphen-A) are that the one kind for finding in oncology and immunological investigation in recent years can suppress high The compound of metabolizing cells growing multiplication, specific can suppress (the Acetyl of acetyl-CoA carboxylase -1 in cellular process Coenzyme (CoA) carboxylase-1, ACC-1) activity,

(Acetyl coenzyme (CoA) carboxylase can be catalyzed acetyl coenzyme A to acetyl-CoA carboxylase (acetyl CoA) is converted into malonyl coenzyme A (malonyl CoA).Malonyl coenzyme A can regulate and control the life of LCFA Thing synthesizes and degrades.There are the ACC of two kinds of hypotypes, i.e. ACC1 and ACC2 in vivo.ACC1 is predominantly located in cytoplasm, and ACC2 is main Positioned at mitochondria.The malonyl coenzyme A catalyzed and synthesized by ACC1 in cytoplasm is the long-chain fat of fatty acid synthetase catalysis The important carbon donor of acid synthesis, action sites of the Suo Lafen-A in cellular process is shown in Fig. 1.And in mitochondrial surface The malonyl coenzyme As that catalyze and synthesize of ACC2 can then suppress carnitine acyl transferase I, enter line so as to regulate and control LCFA Plastochondria carries out beta oxidations.

In recent years, there is increasing research all using ACC1 as a target for being expected to treat metabolic disease and tumour Molecule.Knocking out ACC1 by liver specific genes can be with recombining for effective for fat loss acid and tiring out for triglycerides Product, and fat specific knockdown ACC1 then reduces the sluggishness of Skeletal Muscle Growth.And in metastatic tumo(u)r, ACC1 or aliphatic acid The expression of synzyme has all been increased.

The invention provides a kind of new application of Suo Lafen-A, i.e., for the cerebral protection after ischemic brain damage.

Suo Lafen-A of the present invention can with by pharmaceutically or physiology it is acceptable acid or alkali derived from salt form use. The salt that these salt are including but not limited to formed with following acid：Hydrochloric acid, hydrobromic acid, sulfuric acid, citric acid, tartaric acid, phosphoric acid, breast Acid, pyruvic acid, acetic acid, butanedioic acid, oxalic acid, fumaric acid, maleic acid, oxaloacetic acid, methanesulfonic acid, ethyl sulfonic acid, benzene sulfonic acid or hydroxyl second Sulfonic acid.Other salt include：The salt formed with alkali metal or alkaline-earth metal (such as sodium, potassium, calcium or magnesium), and with ester, carbamic acid The form of " pro-drug " of ester or other routines.

Ischemic brain damage

As used herein, term " ischemic brain damage ", " ischemic cerebrovascular disease " are used interchangeably, and refer to local cerebral Tissue, including denaturation, the downright bad or transient function that nerve cell, spongiocyte and contact fiber occur due to blood supply disorder Lose.It is a kind of common high lethal of clinic, crippling cranial vascular disease.

Generally ischemic brain damage can be due to causing ischaemia after local organization ischemic, postischemic reperfusion or bleeding Cause Deng many reasons.

The ischemic brain damage that Suo Lafen-A of the present invention are suitable for is not particularly limited, and can include any local cerebral ischemia And with or without the brain damage disease of neurological dysfunction.Common ischemic brain damage includes：Cerebral apoplexy, ischemic- Reperfusion injury, transient ischemic attack, neonatal hypoxic ischemic encephalopathy.

In ischemic brain damage, modal pathological manifestations are the destruction of the oedema of neuron, blood-brain barrier, and periphery is exempted from The brain tissue infiltration of epidemic disease cell, the reconstruction of neural blood vessel unit.Therefore, present invention also offers a kind of suppress periphery immunocyte The method of maincenter infiltration.It is demonstrated experimentally that after it application of Suo Lafen-A of the present invention, the periphery of the maincenter of acute phase of cerebral ischemia is exempted from Epidemic disease cellular infiltration is significantly reduced, and blood-brain barrier disruption mitigates.From the point of view of the reparation of chronic phase, Suo Lafen-A can be by suppressing nerve Inflammatory reaction, plays a part of to promote neural blood vessel unit to rebuild.Periphery function of immune system can be caused after cerebral arterial thrombosis Disorder, ischemic Early manifestation be immunocyte excessive activation, infiltrate to brain tissue.After injury the phase there is immune system work( Can decline, cause infection-related complication.Suo Lafen-A can pass through the activation of regulation and control immunocyte, mitigate the immunity in ischemic later stage Function inhibitio, so as to reduce the infection-related complication of post-stroke, improves the prognosis of patients with cerebral apoplexy.

Pharmaceutical composition and application process

Present invention also offers a kind of pharmaceutical composition for treating ischemic brain damage, described pharmaceutical composition In the Suo Lafen-A containing safe and effective amount or its pharmaceutically acceptable salt, and pharmaceutically acceptable carrier.Additionally, this Bright pharmaceutical composition can also be containing the medicine of other treatment ischemic brain damage, such as thrombolytics, neuroprotective agent etc..

Generally, these materials can be formulated in nontoxic, inert and pharmaceutically acceptable aqueous carrier medium, its Middle pH ordinarily be about 5～8, and preferably pH is about 6～8, although pH value can be with the property for being formulated material and disease to be treated Disease and be varied from.The pharmaceutical composition for preparing can be administered by conventional route, including (but being not limited to)： Orally, intramuscular, intravenous, subcutaneous, intracutaneous or local are administered.

When Suo Lafen-A of the present invention are used for such use, can be with one or more pharmaceutically acceptable carrier or figuration Agent mixes, such as solvent, diluent, preferably can be with sterile injectable solution or form of suspension (containing about in isotonic medium 0.05-5% suspending agents) carry out parenteral routes.For example, these pharmaceutical preparations can contain the about 0.01-99% mixed with carrier, The active component of more preferably about 0.1%-90% (weight).

The effective dose of active component used can be with the tight of compound used, the pattern of administration and disease to be treated Weight degree and change, this generally can be adjusted by those skilled in the art according to actual conditions and limited experiment.So And, generally when the compound of the present invention is given daily with the dosage of about 0.5-1000mg/kg the weight of animals, can obtain making us full The effect of meaning, is preferably given with the dosage of 1-4 time daily.For most of large mammal, daily accumulated dose is about For 1-100mg/kg, 2-100mg/kg is preferably about.Can adjust this dosage to provide optimal treatment response.For example, by An urgent demand for the treatment of situation, daily can pari passu increase or decrease dosage.

A kind of concentration or dosage of preferred active component is about daily 1-100mg/kg, preferably 1-50mg/kg, more It is goodly 1-5mg/kg.Preferably, the dosage can be according between mouse known in the art and people's body weight and body surface area Formula converted.

The compounds of this invention or pharmaceutical composition are preferably administered by the approach such as intravenous.Liquid carrier includes：Sterilized water, Polyethylene glycol, propane diols, ethanol, dimethyl sulfoxide (DMSO), nonionic surface active agent etc., if be adapted to active component characteristic and Required specific administration mode.The adjuvant being usually used in pharmaceutical composition is prepared also can advantageously be included, for example anti-corrosion Agent and antioxidant such as vitamin E, vitamin C, citric acid, BHT and BHA.

Being adapted to the medicament forms of injection includes：Aseptic aqueous solution or dispersion liquid and aseptic powder (are used for extemporaneous preparation of sterile Injection solution or dispersion liquid).In all situations, these forms must be aseptic and must be that fluid is arranged with being easy to syringe Go out fluid.Must be under conditions of manufacture and storage stable, and must be able to prevent the pollution of microorganism (such as bacterium and fungi) Affect.Carrier can be solvent or decentralized medium, wherein containing such as water, alcohol (such as glycerine, propane diols and liquid polyethylene glycol), Their appropriate mixture and vegetable oil.

Clinical practice

The invention provides the new indication of Suo Lafen-A clinical practices.Zoopery shows, apply in advance Suo Lafen- After A, mouse is remarkably reinforced to the tolerance degree of cerebral ischemia, can substantially mitigate the cerebral apoplexy order of severity of induction, and In there is the animal of cerebral apoplexy, the progress that Suo Lafen A are capable of effective control acute apoplexy is applied, reduce infarct size, slow down Neuron and angioedema, and also play positive effect in long-term nervous function protection.

Beneficial effect of the present invention

The invention provides new applications of the antineoplastic Suo Lafen-A in ischemic brain damage, can be used as preventative use Medicine, more can apply in acute attack stage and in chronic convalescence, extend the therapeutic time window of the ischemic brain damages such as cerebral apoplexy, It can substantially mitigate the progress of cerebral apoplexy, improve the function of brain neuron, and more favourable propping up is provided to patients with cerebral apoplexy prognosis Hold.

With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention Rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally according to conventional strip Part, such as Sambrook et al., molecular cloning：Laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) described in condition, or according to the condition proposed by manufacturer.Unless otherwise indicated, it is no Then percentage and number are percentage by weight and parts by weight.

Suo Lafen-the A of embodiment 1 have the work of neuroprotective to the Acute ischemia rePerfusion that middle cerebral artery occlusion causes With

Experimental technique：

1) 8-12 week male C57/BL6 wild-type mices (being purchased from Shrek animal used as test company) is selected, 5 are randomly divided into Group, Sham groups (n=8), DMSO groups (n=8), SorA-1 groups (n=8), SorA-5 groups (n=8), Sor-A-25 groups (n=8).

2) each group mouse is given to set up animal model respectively, Sham group mouse row sham-operations, separating blood vessel does not insert line bolt. SorA group mouse rows middle cerebral artery occlusion operation (MCAO) of DMSO and three dosage group.MCAO surgical procedures are as follows：Will After mouse is with 1.5% isoflurane anesthesia, fixation of lying on the back.Exposure neck, median incision about 3cm, blunt separation, exposure tracheae are blunt Property separate both sides thyroid gland, exposure right side nutator and sternohyoideus between trigonum.Internal carotid is separated to drum Bubble, carefully separates and ligatures arteria pterygopalatina.After the external carotid artery distal end of coagulation right side, close neck with the temporary transient folder of micro- artery clamp and always move Arteries and veins, external carotid artery stump is led to outer lower side, is allowed to internal carotid in straight line, and in external carotid artery stump an osculum is cut, No. 8 line bolts of a diameter of 0.23mm are inserted into internal carotid from external carotid artery, is softly slowly pushed ahead to the power that is hampered, by line bolt Ligature jointly with fixing line bolt with external carotid artery, unclamp artery clamp, sew up the incision.Arteria cerebri media opening is now enclosed, is hindered Broken the blood flow of arteria cerebri media, makes MCAO models.Blocking exposes cervical incision after 60 minutes, slowly pulls out Outlet bolt and realizes again Perfusion.

The MCAO models of mouse can cause the infarct of volume parietal cortex and caudate nucleus, the death rate it is low (<10%), it is suitable for length Phase behavioral function is assessed.

In order to reduce physiologic variables to ischemic brain damage more after potential impact, by rectal temperature and life during testing Reason norm controlling is in normal range (NR).The LCBF during ischemic is monitored using the sxemiquantitative of Speckle laser-Dopplers (rCBF).If cortex rCBF declines during mouse MCAO<20%, by it is disallowable outside.

3) bloodstream blocking pulls out line bolt after 60 minutes, realizes that brain blood flow is filled again.After filling 1 hour again, according to packet abdomen is given Chamber is administered：DMSO group mouse give 200ulDMSO；SorA-1 groups give 10% that Suo Lafen-A1mg/kg are dissolved in 200ul In DMSO；SorA-5 groups give Suo Lafen-A 5mg/kg and are dissolved in 10% DMSO of 200ul；SorA-25 groups give Suo La Sweet smell-A 25mg/kg are dissolved in 10% DMSO of 200ul.

4) Post operation 1 day, 2 days and 3 days, determines respectively the scoring of mouse Nerve functional defect, and standards of grading are as follows：It is divided into 0- 5 points：0 point is impassivity dysfunction；1 point is unable to full extension for right side fore paw；2 points is to turn-take to the right；3 points is to roll to the right ；4 points is to walk；5 points are death.

5) Post operation the 3rd day, puts to death mouse, takes Application mouse brain section mould after brain anterior to after hippocampus from caudate nucleus Tissue does continuous coronal section between portion (thickness is 1mm).Brain piece is infiltrated on into the 2%TTC (nitrogen of 2,3,5-chlorinated triphenyl base four Azoles).Because TTC is the proton acceptor of pyridine in respiratory chain-nucleotide structure enzyme system, with the dehydrogenase reaction in normal structure And take on a red color, and dehydrogenase activity declines in ischemic tissue, it is impossible to react, therefore change will not be produced in pale.Therefore TTC dyeing Can be used to dye the ischemic infraction of detection mammalian tissues.Fix overnight using 4% paraformaldehyde after dyeing, second day Can be taken off shooting TTC stained photographs.According to the method for Swanson, based on ischemic side and non-ischemic Ce Wei infarction tissues, analysis is every Infarcted region on brain piece, can avoid that infarcted region is angioedematous to obscure effect.Infarction volume=infarct size summation × All brain piece thickness.

Experimental result：

The brain blood flow situation of change in MCAO surgical procedures is monitored using laser speckle Doppler technology, such as Fig. 2A-B are little After mouse row sham-operation, Bilateral hemispheres brain blood flow is without significant changes；In DMSO groups and SorA group mouse, arteria cerebri media Obstruction cause the significant brain blood flow of side cerebral hemisphere and reduce, and the reduction degree of brain blood flow in this two groups of mouse simultaneously There was no significant difference (Fig. 2 B).The method dyeed by TTC, calculates the volume of ischemic tissue of brain, and it was found that Jing SorA (1, 5th, 25) cerebral infarction volume of the mouse for the treatment of is significantly less than DMSO control mices (Fig. 2 C-D).Therefore, demonstrate,proved by this experiment Bright, SorA has protective effect to Acute ischemia rePerfusion.

Suo Lafen-the A of embodiment 2 can improve nervous function long-term after ischemic brain damage

This experiment employs a series of effective, sensitive, the reproducible behaviouristics detection method of rodents to assess Neurological dysfunction after ischemic brain damage.All appraisal procedures start to enter weekly for 1 week after MCAO Post operations or sham-operation Row test.

1) Corner tests：Corner tests can detect that the sensorimotor function after MCAO to 90 days is damaged.Will experiment Animal is placed in the clamping plate entrance of 30 ゜ so as to voluntarily advance to corner.When mouse advances to angle head end, can to the left or to the right Turn to, then mouse is taken out and is replaced in clamping plate entrance, repeat above step.Each mouse gives 10 advance steering machines Meeting, records the evaluation index that the number of times for turning to the left is tested as corner.

2) wrong step test：For evaluating forelimb and hind leg function.Mouse is placed in 30 (length) × 35 (width) × 31 (height) cm Grid on (grid is 2.5cm²).Mouse steps on steel wire and moves on grid.The claw sky that drops or step on just is designated as once wrong step. Each animal testing 1min, totally 3 times, per minor tick 1min.Calculate the percentage that contralateral limbs mistake step number accounts for total mistake step number.

3) cylinder test (Cylinder Test)：Cylinder test can assess long-term sensation and motor impairment and Limbs symmetry, is detected for postoperative 3 days, 5 days, 7 days, 10 days, 14 days, 21 days and 28 days in this research in mouse.Specifically It is as follows：Animal used as test is put into into diameter 8cm, in the transparent plastic drum of high 15cm, 2-4 animal is placed every time, it is ensured that per only A mouse is placed in cylinder.Carry out and recorded a video 5 minutes after mark.Complete it is all video recording work after, by one to experiment packet not Onlooker person in the know carries out technology to the number of times that upper limbs in mouse in video recording 5 minutes contacts upwards cylindrical wall.Final evaluation index Be expressed as non-damaging side (right side) limbs preference use ratio, i.e. (right side-left side)/(right side+left side+bilateral) frequency of exposure × 100。

4) neural sensation is assessed with motor function：

A) limbs feel function score：Every side limbs of mouse are contacted with a blunt rod, and it is anti-to what is stimulated to observe it Should.Standards of grading are as follows：3rd, stimulation of the mouse to both sides is equally sensitive, and has the reaction of rotary head；2nd, bilateral habituation or right Side reaction weakens；1st, mouse is blunt to the IR in left side；With 0, stimulation of the mouse to left side is reactionless.

B) climbing experiment：Mouse is placed on wire mesh cage.Under normal circumstances, mouse climbs up wire mesh cage with all of four legs Metope.After mouse removes from wire mesh cage, the muscular strength that mouse catches wire mesh cage is recorded.Standards of grading are as follows：3rd, mouse is light Climb up, hold on to wire netting；2, left side is damaged, and left side muscular strength is weaker than right side；1, mouse cannot climb, or pirouette Circle.

C) forelimb walking experiment：Lift the tail of mouse so as to which hind leg leaves ground, is only walked with forelimb.3rd, trippingly Go ahead and symmetrical；2nd, offset to one side；1st, turn-take；0th, forelimb movements can not.

D) side turns experiment：Mouse is raised more than 10 centimetres from the desktop sides to observe mouse in the air and turns ability, scoring Standard is as follows：3. turn to bilateral equality side, and 45 degree of side gyration ＞；2. turn to bilateral equality side, but side gyration<45 degree； 1st, unequal side turns；0th, turn without side.

E) overall neurological function score：The scoring by more than is added and obtains.

F) sticky paper experiment：Sticky paper is sticked in the centre of the palm of the forelimb of mouse, and determine mouse feel after the presence of sticky paper it Any location contacts of limbs remove the required time to the time of sticky paper and by sticky paper.If mouse still cannot remove in 120s Sticky paper, then the maximum duration that record removes sticky paper is 120s.

5) Morris water maze tests assess the obstacle of long-term cognitive function：The impaired learning and memory of detection MCAO mouse Function：Mouse finds hiding platform so as to flee from the swimming task being forced in open Morris water mazes learning.It is postoperative Mouse was placed in the water maze without platform in 13 days allows it to adapt to, altogether twice.Hereafter, daily water-filling in the 2nd, 4,6 and 8 week after surgery Labyrinth is tested, four times a day.Record animal reaches the required time (incubation period) of submerged platform.When finally training twice, remove flat Platform, allows mouse arbitrarily to swim 30 seconds, and records mouse mutually limiting (the mutually limit that platform was once placed) the interior time of staying and swimming in target Speed.

As a result

1) Corner tests：Advance at 10 times of mouse and turn in chances, the cerebral ischemia mouse of Jing Sor-A treatments to The ratio of left steering is better than control mice less than the ratio of DMSO groups, the limbs symmetry for representing treatment mouse.See Fig. 3.

2) wrong step test：Compared with DMSO mouse, the cerebral ischemia mouse offside forelimb and hind leg mistake step number of Jing Sor-A treatments The percentage for accounting for total mistake step number is significantly reduced, and is close to sham groups mouse.See Fig. 4.

3) cylinder test：After injury, non-damaging side (right side) limbs preference use ratio significantly rises cerebral ischemia mouse Height, the preference use ratio of Sor-A treatment mouse limbs is substantially less than DMSO control groups.See Fig. 5.

4) neural sensation is assessed with motor function：

A) limbs feel function score：The cerebral ischemia mouse of Sor-A treatments damages offside and the reaction for stimulating is significantly higher than DMSO groups, see from.

B) climbing experiment：After cerebral ischemia, the climbing ability of mouse is significantly reduced, Jing Sor-A treatments, its climbing ability Higher than DMSO groups, Fig. 6 B are seen.

C) forelimb walking experiment：The forelimb locomotor activity of mouse is remarkably decreased after cerebral ischemia, and it is little that Jing Sor-A are treated Mouse fall reduces, and sees Fig. 6 C.

D) side turns experiment：Mouse is raised more than 10 centimetres from the desktop sides to observe mouse in the air and turns to be found during ability Compared with DMSO control group mices, its side turns ability and is significantly increased the mouse of Jing Sor-A treatments, sees Fig. 6 D.

E) after above-mentioned scoring is added up, the comprehensive grading of mouse shows, Sor-A treatments are remarkably improved the neural work(of mouse Energy comprehensive grading, is shown in Fig. 6 E.

F) sticky paper experiment：After cerebral ischemia, the time needed for the sticky paper in the centre of the palm is removed dramatically increases mouse, Sor-A The time that treatment mouse is removed needed for sticky paper significantly reduces, and sees Fig. 6 F.

5) Morris water maze tests：Mouse is notable after cerebral ischemia in the time of staying of target quadrant in water maze Reduce, Sor-A treatments significantly increase its time of staying in target quadrant.And the swimming rate of three groups of mouse does not have conspicuousness Difference.See Fig. 7.

In sum, behaviouristics testing result shows that Suo Lafen A can improve sensation long-term after ischemic brain damage, fortune It is dynamic, limbs symmetry and learning and memory function.

Suo Lafen-the A of embodiment 3 can prevent the nervous function damage of (or ischaemic neuronal) after ischemic brain damage

8-12 week male C57/BL6 wild-type mices (being purchased from Shrek animal used as test company) is selected, 3 groups are randomly divided into, Sham groups (n=8), DMSO groups (n=8), SorA groups (n=8).SorA groups give 1mg/kg/d, and lumbar injection is pre-processed 3 days It is followed by being performed the operation by Sham or MCAO.

Postoperative 3 days, 5 days, 7 days, 14 days, row Animal Behavior Science detection in 28 days.Method is ibid.Put to death mouse within 3 days after surgery, Perfusion takes and carry out after brain NeuN immunofluorescence dyeings, determines the existing state of neuron.

Test result indicate that, the neuron of survival is significantly more than in the ischemic area of the mouse that application of Suo Lafen-A in advance DMSO control group mices, are shown in Fig. 8.

The dose screening experiment of the Suo Lafen-A of embodiment 4

The same mouse of this experiment Example 1, and mouse is divided into into 6 groups；N=8, the Suo Lafen A dosage point of per group of employing It is not：G0 placebos are used as control；G1 Suo Lafen-A 0.1/kg mg；G2 Suo Lafen-A 1/kg mg；G3 Suo Lafen-A 5/kg mg；G4 Suo Lafen-A 25/kg mg；G5 Suo Lafen-A 50/kg mg.G0-G5 is repeated the experiment content of embodiment 1-3.

As a result (not shown) shows, when the mouse dose of Suo Lafen A is 0.1 or 1/kg mg, Suo Lafen A to acute or Chronic ischemic brain damage has certain effect, but without significant difference compared with control G0 groups；

When Suo Lafen A mouse dose be 5,25,50/kg mg when, Suo Lafen A are to formulation or chronic ischemic brain damage Effect (kinesitherapy nerve, sensory nerve have significantly recovery) is significantly improved, but the result between G3, G4, G5 is without aobvious Write sex differernce.

Therefore, Suo Lafen A but work as in low dose for the nerve recovery after ischemic brain damage has a faint effect When dosage rises to 5mg/kg, then start to produce obvious action, and effect does not substantially rise after escalated dose.

All experiment mices do not produce obvious or serious bad reaction.

The all documents referred in the present invention are all incorporated as in this application reference, independent just as each document It is incorporated as with reference to such.In addition, it is to be understood that after the above-mentioned instruction content for having read the present invention, those skilled in the art can To make various changes or modifications to the present invention, these equivalent form of values equally fall within the model that the application appended claims are limited Enclose.

Claims (10)

1. the purposes of Suo Lafen-A (Soraphen-A) or its pharmaceutically acceptable salt, it is characterised in that pre- for preparing (i) Pharmaceutical composition that is anti-and/or treating ischemic brain damage；And/or (ii) protects the pharmaceutical composition of ischaemic neuronal, its In, the Suo Lafen-A have structure shown in Formulas I：

2. purposes as claimed in claim 1, it is characterised in that described ischemic brain damage includes Acute ischemia rePerfusion Or chronic ischemic brain damage.

3. purposes as claimed in claim 2, it is characterised in that the Acute ischemia rePerfusion includes that disease time is 2 little When -3 days between ischemic brain damage.

4. purposes as claimed in claim 2, it is characterised in that the chronic ischemic brain damage include disease time 3 days it Ischemic brain damage afterwards.

5. purposes as claimed in claim 1, it is characterised in that described ischemic brain damage includes cerebral apoplexy, transient ischemia/reperfusion Note damage, transient ischemic attack, neonatal hypoxic ischemic encephalopathy.

6. purposes as claimed in claim 1, it is characterised in that the application dosage of the Suo Lafen-A is 1-100mg/kg/ days.

7. purposes as claimed in claim 1, it is characterised in that described pharmaceutical composition contain Suo Lafen-A or its pharmaceutically Acceptable salt is used as active component, and pharmaceutically acceptable carrier.

8. purposes as claimed in claim 7, it is characterised in that described pharmaceutical composition also containing thrombolytics, rtPA and/or Neuroprotective agent.

9. a kind of method that external non-therapeutic protects ischaemic neuronal, it is characterised in that including step：Drawing containing rope Ischaemic neuronal is cultivated under conditions of sweet smell-A or its pharmaceutically acceptable salt, and/or to the neuron of Jing ischemics process Suo Lafen-A are added in culture, so as to protect ischaemic neuronal.

10. method as claimed in claim 9, it is characterised in that described protection includes that preventive protection and/or therapeutic are protected Shield.