WO2005021481A1 - N, n’-dibenzyl ethylenediamine salt pf 2-(alpha-hidroxypentyl) benzoic acid and its preparing process and usage - Google Patents

Abstract

Provided is, N,N’-dibentzyl ethylenediamine salt of 2-(alpha-hydroxypentyl) benzoic acid, which has significant effect in inhibitting blood platelet aggregation and improving cerebral circulation, the pharmaceutical activity against cardioischemia, cerebroischemia, and cardiac or cerebral arterial embolism, good appearance, physical state and wet stability. The preparing process of the salt, the pharmaceutical composition containing it as active component and its use in preparing medicine against cardioischemia, cerebroischemia, cardiac or cerebral arterial embolism and the like are also disclosed.

Description

 2- (c-hydroxypentyl) benzoic acid N, N, -dibenzylethylenediamine salt

 And preparation method and application thereof

 The present invention relates to a novel 1- (α-hydroxypentyl) benzoic acid salt, in particular to 2_ (α-hydroxypentyl) benzoic acid N, N, -dibenzylethylenediamine salt, and The preparation method and the pharmaceutical composition using the salt as an active ingredient also relate to the application of the salt in the preparation of a medicament for treating cardio-cerebral ischemia, cardiac artery obstruction and the like. Background technique

 Acute ischemic stroke, coronary heart disease, and myocardial infarction are all diseases that are caused by atherosclerosis until the formation of thrombus, which leads to ischemic injury. Such diseases bring great pain and even life threat to patients. At present, research on drugs for treating such diseases has been a hot spot and frontier of drug research and development.

 Chinese patent CN1382682A, application number 01109795. 7, first published 2-(cc

-Hydroxypentyl) benzoate, and its preparation method and use, which relate to monovalent metal ions, divalent metal ions and salts of organic bases, and specifically disclose potassium, sodium, calcium, magnesium, zinc, aniline, benzylamine , Morpholine, diethylamine salt. The specification only discloses the preparation of potassium salt, sodium salt, lithium salt, calcium salt, and benzylamine salt, but does not disclose specific preparation methods of zinc salt, aniline, morpholine, and diethylamine salt. The specification also discloses the effects of potassium salts on cerebral infarction area in rats with ischemic cerebral ischemia, platelet aggregation in rats, and the protective effect of isolated heart ischemia-reperfusion arrhythmia in rats. The above experiments have played a beneficial role. Other salts have not been tested for efficacy.

 When the inventors of the present invention developed a new medicine for treating cardiovascular and cerebrovascular diseases, they researched a large number of published 1- (α-hydroxypentyl) benzoic acid salts, and found salts of the prior art (including Chinese patent CN1382682A). There are the following deficiencies in its preparation method:

1) Some bases cannot react at all, such as aniline and similar compounds, because aniline compounds are too basic; 2) Although some bases can react, they can't get solids, including simple organic amines or inorganic ammonia, such as ammonia, amidine, diamine, diethylamine, ethylenediamine, triethylamine, etc., and simple Nitrogen-containing heterocyclic compounds, such as morpholine and piperidine, can only obtain viscous oils;

 3) Some bases are prepared according to the existing published preparation methods. The solid obtained is amorphous, has no fixed melting point and good appearance and physical form, such as sodium salt, magnesium salt, lithium salt, and zinc salt. After the dry solvent, it is a foamy solid or gel. It has no crystalline form, is very viscous, is difficult to pulverize, can not talk about fluidity, and is not suitable for processing by the pharmaceutical industry.

 4) Although some solids can get good solids after reaction, they are very toxic, such as benzylamine and tert-butylamine salts.

 5) Although some inorganic salts (such as potassium salts) have good crystal morphology, they are highly hygroscopic, unstable to humidity, and easy to decompose when placed under ordinary circumstances. In addition, potassium salts appear to be more effective when administered intravenously. Great toxicity. Summary of the invention

 The purpose of the present invention is to provide a new 2- (α-hydroxypentyl) phenyl sulfonate which can overcome the shortcomings of the prior art, which has obvious anti-platelet aggregation and improvement of cerebral circulation, and has good effects on the heart and brain. Ischemia and cardio-cerebral artery occlusion have drug activity, have good appearance and physical form, have no hygroscopicity, and have excellent wet stability.

 The novel salt of the present invention is 2- (α-hydroxypentyl) phenylarsinic acid N, N'-dibenzyl ethylenediamine salt, and its structural formula is as follows:

本发明的盐具有如下优异的性能：

The salt of the present invention has the following excellent properties:

1) It has good crystal morphology and stable melting point, and can be used for pharmaceutical purity by simple post-treatment; 2) Non-hygroscopic, stable to humidity, excellent physical appearance, suitable for storage and processing in the pharmaceutical industry;

 3) Animal acute toxicity test shows that the salt of the present invention is less toxic than the published 1- (ex-hydroxypentyl) benzoate;

 4) In animal model tests, compared with the published 1-(ex-hydroxypentyl) phenylarsinate, it shows more excellent protection against brain tissue damage caused by cerebral artery occlusion at the same dose. And antiplatelet aggregation. It shows that the salt of the present invention has better preventive and therapeutic effects on heart and brain anemia, and has anti-platelet aggregation, treats heart and cerebral artery obstruction, and improves pharmacological effects of heart and brain: circulation;

 5) In animal model tests, compared with the published 2- (ex-hydroxypentyl) phenylarsinate, it has a smaller significant therapeutic dose, that is, the salt of the invention at a lower dose You can show a significant effect. Further considering that the molecular weight of this salt (656.86, potassium salt 246.35, calcium salt 454.57) is large, this product can show a significant effect at a smaller dose in moles. .

 Another object of the present invention is to provide a method for preparing the above 2-(α-hydroxypentyl) phenylarsinic acid N, N, -dibenzylethylenediamine salt, which comprises the following steps:

 (1) The acidification operation is performed on 2-(α-hydroxypentyl) benzoate, and the temperature of the acidification process is controlled-10 ~ 1 (TC, pH 2-3, then extracted with an organic solvent, and the organic solvent layer is washed to neutrality Then, it is dried, and the organic solvent is removed under reduced pressure to obtain 2-(α-hydroxypentyl) phenylarsinic acid;

 (2) The 2- (α-hydroxypentyl) phenylarsinic acid in step (1) is dissolved in an organic solvent, and a solution prepared by adding N, N, -dibenzylethylenediamine in an organic solvent is reacted to control, The reaction temperature is-10 ~ 10 ° C;

 (3) The reaction product in step (2) is stirred for several hours and then filtered, the filter cake is washed with an organic solvent, and the filter cake is dried to obtain 2-(c-hydroxypentyl) phenylarsinic acid N, N, -di Benzylethylenediamine salt.

In the above preparation method, the organic solvent may be diethyl ether, dichloromethane or ethyl acetate, and preferably diethyl ether. In the above preparation method, the reaction temperature described in step (2) is preferably 0 to -5 ° C. Another object of the present invention is to provide 2- (α-hydroxypentyl) phenylarsinic acid N, N'-dibenzylethylenediamine salt for preparing and / or treating cardio-cerebral ischemia and / or cardio-cerebral artery Obstruction and improvement of the brain ^: Application of drugs in circulation.

 A further object of the present invention is to provide a pharmaceutical composition capable of preventing and / or treating cardio-cerebral ischemia and / or cardio-cerebral artery occlusion and improving cerebral microcirculation, which contains a therapeutically effective amount of 2-(a -Hydroxypentyl) phenylarsinic acid N, N,-Dibenzylethylenediamine salt is an active ingredient, and may also contain one or more pharmaceutically acceptable carriers.

 The above pharmaceutically acceptable carriers refer to conventional pharmaceutical carriers in the pharmaceutical field, fillers such as starch, sucrose, etc .; binders such as cellulose derivatives, alginates, gelatin, and polyvinyl pyrrolidone; wetting agents such as ethanol, water Disintegrating agents such as starch and its derivatives, low-substituted hydroxypropyl cellulose, effervescent disintegrating agents; absorption enhancers such as quaternary ammonium compounds; surfactants such as cetyl alcohol; adsorption carriers such as kaolin and soap clay; Lubricants such as talc, calcium and magnesium stearate, and polyethylene glycol; in addition, other adjuvants such as flavoring agents, sweeteners, etc. may be added to the composition.

 The various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional production methods in the pharmaceutical field. For example, the active ingredient is mixed with one or more carriers and then made into the desired dosage form, including tablets, capsules and granules. detailed description

The following examples will further explain the present invention, but the present invention is not limited to these examples, and these examples do not limit the scope of the present invention in any way. Certain changes and adjustments made by those skilled in the art within the scope of the claims should also be considered to belong to the scope of the present invention. Example 1: Preparation of 2- (α-hydroxypentyl) benzoic acid N, N, -dibenzylethylenediamine salt In this example, 2- (ex-hydroxypentyl) benzene was prepared according to the following four schemes, respectively. The acetic acid N, N, -dibenzylethylenediamine salt, the specific operation process is as follows. Option A

 Preparation of α-hydroxy-n-pentylbenzoic acid

将 α-羟基正戊基苯曱酸钠 (按 CN1382682Α制备 )80 g( 0.348mol ) 和水 500ml加入到 2.5L的烧杯中， 搅拌溶解后冷至 -1 (TC， 分批加入 冷的 5%的盐酸， 控制内温在 -5 至 -1(TC, 调 PH=2.0， 用乙醚 500ml x 3萃取， 用水 300ml X 3洗乙醚层， 洗至中性， 乙瞇层干燥， 滤除 干燥剂， 减压将乙醚除尽， 得白色固体 64g, 收率 88.4%;

Add 80 g (0.348mol) of α-hydroxy-n-pentyl benzoate sodium (prepared according to CN1382682A) and 500ml of water to a 2.5L beaker, stir to dissolve and cool to -1 (TC, add cold 5% in portions) Hydrochloric acid, control the internal temperature to -5 to -1 (TC, adjust the pH = 2.0, extract with 500ml x 3 ether, wash the ether layer with water 300ml x 3, wash until neutral, dry the ethyl acetate layer, filter off the desiccant, The ether was removed under reduced pressure to obtain 64 g of a white solid with a yield of 88.4%;

 Preparation of 2- (α-hydroxy-n-pentyl) phenylarsinic acid N, N, -dibenzylethylenediamine salt

Add 37.2g (0, 155mol) of Ν, Ν, -dibenzylethylenediamine and 1.0L of diethyl ether to the reaction flask. After being completely dissolved under stirring, 2- (α-hydroxy-n-pentyl) phenylarsine is added in portions. 1.0L (64g, 0.308mol) of diethyl ether solution, the reaction was controlled at (TC ~ -5 ° C), gradually became cloudy, and a large amount of white solid precipitated after the addition;

After stirring for 3-4 hours, suction filtration was performed. The filter cake was washed with 1500 ml of ether. Transfer to a 2.5L beaker, wash twice with 2.0L of diethyl ether, filter, and put the filter cake in a vacuum dryer to obtain 92 g of a white solid, mp: 105-106 ° C, yield: 91.0%.

 Option B

 (1) Add 86 g (0. 350mol) of a-hydroxy-n-pentylphenylbenzoate (prepared according to CN1382682A) and 500ml of water to a 2.5L beaker, stir to cool to -5

° C, add cold 5% hydrochloric acid in batches, control the internal temperature at 0 to -5 ° C, adjust the pH = 2.5, extract with 500ml X 3 of dichloromethane, wash the dichloromethane layer with 300ml X 3 water, Wash until neutral, dry the dichloromethane layer, filter off the desiccant, remove the dichloromethane under reduced pressure to obtain 61 g of a white solid; yield 83.8%

 (2) Add 34.6 g (0.144 mol) of N, N, -dibenzylethylenediamine and 1.0 L of dichloromethane into the reaction flask, and after dissolving, add 2- (α-hydroxy-n-pentyl) in portions. Base) Benzoic acid in dichloromethane solution 1.0L (60g, 0.288mol), the temperature was controlled at -10 ° C ~ -5 ° C to react, gradually became turbid, and a large amount of white solid precipitated after the addition;

 (3) After stirring for 3-4 hours, suction filtration is performed. The filter cake is washed with 1500 ml of dichloromethane. After drying, the solid is transferred to a 2.5 L beaker, washed twice with 2.0 L of dichloromethane, filtered, and the cake is filtered. Put it into a vacuum dryer to obtain 83.3 g of white solid, mp: 103-105 ° C, yield: 88.2%.

 Option C

 (1) Add 80 g (0.176 mol) of α-hydroxy-n-pentylphenyl arsenate and 500 ml of water to a 2.5 L beaker, cool to -5 ° C, and suspend without completely dissolving, stirring Add cold 5% hydrochloric acid in batches, control the internal temperature at 0 to -5 ° C, adjust the pH to 3.0, extract with 500ml of ethyl acetate x 3, wash the ethyl acetate layer with 300ml of water, and wash until neutral. The ethyl acetate layer was dried, and the desiccant was filtered off. The ethyl acetate was removed under reduced pressure to obtain 62 g of a white solid in a yield of 84.6%;

(2) Add 34.6 g (0.144 mol) of N, N'-dibenzylethylenediamine and 1.0 L of ethyl acetate to the reaction flask, and completely dissolve under stirring, and then add 2- (α-hydroxy-n-pentyl) in portions. ) 1.0 L (60 g, 0.288 mol) of phenylarsinoic acid in ethyl acetate, the reaction was controlled at a temperature of 0 ° C to 10 ° C, and gradually became cloudy, and a large amount of white solid precipitated after the addition; (3) After stirring for 3-4 hours, suction filtration is performed. The filter cake is washed with 1500 ml of ethyl acetate. After drying, the solid is transferred to a 2.5 L beaker, washed twice with 2.0 L of ethyl acetate, filtered, and the filter cake is placed. Dry in a vacuum dryer to obtain 78.7 g of a white solid, mp: 105-106 ° C, yield: 83.3%.

 Option D

 (1) Add 108 g (0.350 mol) of α-hydroxy-n-pentyl phenylarsinic acid salt (prepared according to CN1382682A) and 500 ml of water to a 2.5 L beaker, stir to dissolve and cool to -10 ° C, batch by batch Add cold 5% hydrochloric acid, control the internal temperature at 0 to -5 ° C, adjust the pH to 2.4, extract with 500ml X 3 ether, wash the ether layer with 300ml X 3 water, wash to neutral, dry the ether layer, filter off Desiccant, diethyl ether was removed under reduced pressure to obtain 64g of a white solid with a yield of 88.0%;

 (2) Add 37.2 g (0.155 mol) of N, N, -dibenzylethylenediamine and 1.0 L of diethyl ether to the reaction flask, and after dissolving in the solution, add 2- (ex-hydroxy-n-pentyl) benzene in portions. 1.0 L (64 g, 0.308 mol) of acetic acid in diethyl ether, the temperature was controlled at 0 ° C ~-5 ° C, the reaction gradually became cloudy, and a large amount of white solid precipitated after the addition;

 (3) After stirring for 3-4 hours, suction filtration is performed. The filter cake is washed with 1500 ml of diethyl ether. After suction drying, the solid is transferred to a 2.5 L beaker, washed twice with 2.0 L of ether, filtered, and the filter cake is dried in a vacuum dryer. 92.5 g of white solid was obtained, mp: 105-106 ° C, yield: 91.5%.

 Option E

(1) 77 g (0.405 mol) of racemic 3-n-butylisobenzofuran-1- (3H) -one and 200 ml of methanol were added to a 500 ml reaction bottle, stirred and heated to reflux, and 70 ml of NaOH aqueous solution was added in portions. (Containing NaOH28g, 0.7mol), reflux for 8 hours after completion of the addition, evaporate under reduced pressure, remove all alcohol, add 300ml of water, stir and dissolve the reaction solution with 300ml of ether, and cool the aqueous phase to -10 ° C. Add cold 5% hydrochloric acid in batches, control the internal temperature at 0 to -5 ° C, adjust the pH to 2.7, extract with 500ml x 3 ether, wash the ether layer with 300ml x 3 water, wash to neutral, dry the ether layer, filter Remove the desiccant, cool to -10 ° C ~ -5 ° C in a 5L reaction flask, and add the N, N, -dibenzylethylenediamine solution (37.2g, 0.155mol, dissolved in 1.0L ether) while stirring. ), Control the temperature at 0 ° C ~-5 ° C for reaction, gradually become turbid, after the addition of a large number of white solids; (2) After stirring for 3-4 hours, suction filtration is performed. The filter cake is washed with 1500 ml of diethyl ether. After suction drying, the solid is transferred to a 2.5 L beaker, washed twice with 2.0 L of ether, filtered, and the filter cake is placed in a vacuum dryer. Dry to give 93 g of white solid, mp: 105-106 ° C. Analysis of 2- (α-hydroxy-n-pentyl) benzoic acid N, N, -dibenzylethylenediamine salt prepared according to the above five schemes, the results obtained are as follows:

Infrared absorption spectrum, KBr, 3419cm ^-1 '(γ _N „), 3040, 3070 cnf ¹ ′ (γ _ΑΓ- „). 1763 cm " ^1, (γ co)

NMR proton spectrum, (300MHz, CD ₃ C1) δ (ppm) 7.13-7.60 (m, 14H), 4.93 (t, 2H, two Ar-CH), 3.92 (S, 4H, two Ar-CH ₂ ), 2.99 (s, 4H, N-CH ₂ CH ₂ -N), 1.63-1.70 (m, 4H, two CH ₂ ), 1.14-1.36 (m, 8H, two CH ₂ CH ₂ ) 0.76- 0.83 (m, 6H, two CH ₃ )

ESI-MS:

Negative M / Z 207

r,

Positive M / Z 241

Elemental analysis: C ₄ . H ₅₂ N ₂ 0 ₆ FW656.86

 C H N

 Theoretical value (%) 73.14 7.98 4.26

Measured value (%) 73.10 8.05 4.28

 **: Prepared by the calcium salt method in CN1382682A

 ***: Prepared according to the method of the scheme of Example 1

 The results show that: lithium salt, sodium salt, magnesium salt, and zinc salt are all foam-like or gel-like solids with no obvious melting point. Morpholine and diethylamine salts are viscous liquids. None of them meet the requirements of the pharmaceutical industry. Potassium, calcium, benzylamine, N, N, -dibenzylethylenediamine and tert-butylamine salts have good physical appearance and fixed melting point. Aniline does not react at all. Example 3: Comparison of wet stability

 The test was performed in accordance with the method described in the "High Humidity Test Method" under "Guidelines for the Stability of Drugs" under Appendix XIX C of the Second Edition of the Chinese Pharmacopoeia 2000 for 10 days. The results are shown in Table 2.

 Table 2 Comparison of the wet stability of potassium salt, calcium salt, benzylamine salt, N, N, -dibenzylethylenediamine salt and tert-butylamine salt

* Decomposition products were determined by HPLC method, according to the VD test in Appendix 2 of Chinese Pharmacopoeia 2000, C18 column, mobile phase acetonitrile: 0.02M sodium dihydrogen phosphate = 40: 60, detection wavelength 230nm, flow rate 1ml / min. The experimental results show that the potassium and benzylamine salts are more hygroscopic and decompose severely after moisture absorption. The N, N, -dibenzylethylenediamine and calcium salts are less hygroscopic and stable. Tert-butylamine salt is not very hygroscopic, but its stability is slightly worse. Example 4: Acute toxicity test

This embodiment uses:

 1.Animals

 ICR mice, male and female, weighing 18-20 g, were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The certificate number is: SCXK (Beijing) 2002-0003.

 Test drug

 Oral: N, N, -Dibenzylethylenediamine salt, potassium salt, calcium salt, benzylamine salt, tert-butylamine salt, all provided by Beijing Tianheng Pharmaceutical Research Institute, with 0.5% carboxymethyl cellulose sodium solution Suspension, formulated in different concentrations; 'Intravenous injection: potassium salt, formulated with saline for injection.

 3. Test method

Sixty ICR mice were randomly divided into six groups according to body weight and sex, with ten in each group, half male and half female. Based on the pre-test results, the doses of each group were designed according to the group spacing 1: 0.8. The oral administration volume is 0.2 ml / 10 g body weight (the potassium salt intravenous administration volume is 0.1 ml / l Og body weight), fasting and water ca n’t be avoided for 12 hours before the test, and conventional feeding after the administration, continuous Observe for 14 days, record the toxic reaction and death of the animals, necropsy was performed in the dead group, pathological changes were observed with the naked eye, and LD ₅ was calculated by Bliss method. Value and 95% confidence limit.

 4. Test results

 Table 3 Acute toxicity test results

Salt name LD ₅ . 95% confidence limit

N, N, -Dibenzylethylenediamine salt 3. 1910g / kg 2. 7851-3. 656g / kg Calcium salt 1. 5498g / kg 1. 3015-1. 8546g / kg Benzylamine salt 1. 4537g / kg 1. 316-1. 6057g / kg tert-butylamine salt 1. 2635 g / kg 1. 1188-1. 427g / kg potassium salt (oral) 1. 1735 g / kg 1. 0559-1. 3041 g / kg potassium salt ( Intravenous injection) 0. 382g / kg 0. 351 3-0. 4154g / kg The results show that the acute toxicity (oral) of each salt to mice is N, N, in that order, Ethylenediamine salt <calcium salt <benzylamine salt <tert-butylamine salt <potassium salt, of which N, N, -dibenzylethylenediamine salt has the least toxicity. Example 5: Effect on Middle Cerebral Artery Thrombos is (MCAT) Model Rats with Neurological Symptoms and Cerebral Infarction

This embodiment uses:

I. Animals

 SD rats, male and female, weighing 190-210g, provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., certificate number: SCX 11- 00-0008.

Drugs and reagents:

 Test drugs: N, N,-dibenzylethylenediamine salt, potassium salt, calcium salt, all provided by Beijing Tianheng Pharmaceutical Research Institute, suspended with 0.5% carboxymethyl cellulose sodium solution, formulated different density.

Reagent: FeC l ₃ · 6H ₂ 0 (AR), produced by Beijing Chemical Plant, formulated with lmo l / L hydrochloric acid; red tetrazolium (TTC), produced by Beijing Chemical Plant.

Third, the instrument

 XTT solid microscope: produced by Beijing Electro-optical Scientific Instrument Factory; constant temperature water bath oscillator SHZ-22 type: produced by Jiangsu Okura Medical Instrument Factory; electronic analytical balance: AEG-220 type, produced by Shimadzu Island.

4. Test methods and results

 (I) Effects on neurological symptoms in MCAT rats

1. Grouping and administration: Animals were randomly divided into 14 groups, namely the operation group, the MCAT model group, N, N, -dibenzylethylenediamine salt 340mg / kg, 200mg / kg, 170mg / kg, 100mg / kg, 85mg / kg, 50mg / kg group, potassium salt 200mg / kg, 100mg / kg, 50mg / kg group, calcium salt 200mg / kg, 100mg / kg, 50mg / kg group. Preventive administration before modeling, intragastric administration once a day, continuous administration for 3 days, on the 4th day, modeling, intragastric administration immediately after the model is given a set dose, 12 hours after modeling, the stomach is given once again. Fixed dose. The control group was given the same amount of solvent. 2. Modeling operation method: Rats are anesthetized by intraperitoneal injection of 12% chloral hydrate solution (350mg / kg). Refer to the method of Tanrnra et al. (Tamura A, Graham DI, McCulluoch J et al. Focal cerebral ischemia in the rat. 1. Description of technique and early neuropathological consequences following middle Cerebral artery occlusion. J Cereb Blood Flow Metab, 1981, 1: 53 ) And improved, the rat was placed in the right lateral position, and an arc-shaped incision was made at the midpoint of the conjunctiva of the eye and the external auditory canal, approximately 1.5 cm long. The temporal muscle was clipped and cut away to expose the temporal bone. A dental drill was used. A bone window with a diameter of 2.5 mm was made at the confluence of the zygomatic bone and the temporal squamium to the mouth 1 to clear the residue and expose the middle cerebral artery (located between the olfactory tract and the inferior cerebral vein). A small quantitative filter paper sucked with 50% ferric chloride solution 10 μl was applied to this middle cerebral artery. After the color of the blood vessels became black for about 30 minutes, the filter paper was removed, and the local tissue was washed with physiological saline, sutured layer by layer, and returned to the cage for feeding.

 3. Behavior detection: Refer to the method of Bederson et al. (Bederson JB, Pitts LH, Tsuji M et al. Rat middle cerebral artery occlusion: evaluation of the mode and development of a neurologic examination. Stroke, 1986, 17: 472) and improve it. Animals were scored at different time points (6h, 24h) after operation. (1) Lift the rat's tail about one foot away from the ground and observe the forelimb flexion. If both forelimbs are symmetrically extended to the ground, 0 points are scored; if the contralateral forelimbs on the contralateral side of the surgery have shoulder flexion, elbow flexion, shoulder rotation or those with both wrist and elbow flexion and internal rotation, 1 point is scored. (2) Place the animal on a smooth ground, push the shoulders to the opposite side, and check the resistance. If the bilateral resistance is equal and strong, it is scored as 0 points; if the resistance decreases when pushed to the opposite side of the operation, it is scored as 1 point. (3) Place the two forelimbs of the animal on a metal net, and observe the muscle tension of the two forelimbs. Bilateral muscle tension is equal and strong is 0 points; the contralateral forelimb muscle tension drop is recorded as 1 point. (4) The mouse tail is about one foot away from the ground, and the animal keeps rotating toward the opposite side of the operation, which is recorded as 1 point. Based on the above criteria, the maximum score is 4 points. The higher the score, the more severe the animal's behavioral disorder.

4. Analysis of results: The t-test was used to compare the behavior detection scores between groups. The results are shown in Table 4. Effect on neurological symptoms in MCAT rats

 Note: Compared with the model group, *** P <0. 001, ** P <0. 01, * P <0

The results showed that the behavior of the sham operation group showed no abnormal changes. The rats in the model group showed hemiplegia-like symptoms at 6 and 24 hours after operation. The main symptoms were adduction of the contralateral forelimbs, internal rotation of the shoulders, reduction of forelimb muscle tension, and reduction of shoulder resistance. . Compared with the model group, the neurological symptoms of rats in the N, N, -dibenzylethylenediamine salt 100-340mg / kg group were significantly improved at 6 hours after operation. No significant improvement in symptoms (P>0.05); 24 hours after surgery, N, N, -dibenzylethylenediamine salt 50-340mg / kg group, potassium and calcium salts 100mg / kg, 200mg / kg group Neurological symptoms in rats were significantly improved, and there was a significant dose-effect relationship. There was no significant improvement in the potassium and calcium 50 mg / kg group (P> 0.05). The significant therapeutic dose of N, N, -dibenzylethylenediamine salt to improve neurological symptoms in MCAT rats is 50mg / kg, which is lower than potassium and calcium salts (100mg / kg). At the same dose, N, N, -dibenzylethylenediamine salt improved the neurological symptoms of MCAT rats more than potassium and calcium salts. (B) the effect on cerebral infarction range in MCAT rats

The animals were decapitated and brain removed after the last behavioral score. Remove the olfactory bulb, cerebellum and lower brainstem, and cut the remaining part into 5 pieces at 4 ° C. Quickly place the brain slices in TTC staining solution (4% TTC 1.5ml, 1M K ₂ HP0 ₄ 0.1ml per 5ml), incubate at 37 ° C in the dark for 30 minutes, then remove and place in 10 Store in dark solution in% formaldehyde solution. The non-ischemic area was rose red and the infarcted area was white after staining. The white tissue was carefully mined and weighed, and the percentage of infarcted tissue weight to total brain weight was used as the range of cerebral infarction. The t-test was used to compare the results between the groups. The results are shown in Table 5.

 Table 5 Effects on cerebral infarction range in MC AT rats

 Note: Compared with the model group, *** ρ <0. 001, ** ρ <0. 01, * ρ <0. 05

The results showed that the rats in the model group had obvious infarcts 24 hours after the middle cerebral artery infarction in the rats, and there was no change in the sham operation group. Compared with the model group, the range of cerebral infarction in the N, N, -dibenzyl ethylenediamine salt 50-340 mg / kg group, potassium salt, 100 mg / kg, 200 mg / kg group were significantly reduced. And there is a clear dose-effect relationship; potassium and calcium salts 50mg / kg There was no significant inhibitory effect in the groups (P> 0.05). N, N, -Dibenzylethylenediamine salt inhibits the cerebral infarction range of MCAT rats at a significant therapeutic dose of 50 mg / kg, which is lower than potassium and calcium salts (100 mg / kg). _{O At the} same dose, N, Ν ' -The inhibitory effect of dibenzylethylenediamine salt on cerebral infarct size in MCAT rats is stronger than that of potassium and calcium salts. Example 6: Effect on platelet aggregation in rats

 This embodiment uses:

I. Animals

 SD rats, male, weighing 220-250g, provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., certificate number: SCXK11- 00- 0008.

Drugs and reagents

 N, N, -dibenzylethylenediamine salt, potassium salt and calcium salt are provided by Beijing Tianheng Pharmaceutical Research Institute. Positive control drug: Aspirin (ASA) was produced by Mudanjiang Zifeng Chemical Pharmaceutical Co., Ltd., batch number 010622015. The above drugs were suspended in 0.5% sodium carboxymethylcellulose solution and formulated in different concentrations. Adenosine diphosphate (ADP) was purchased from Sigma.

3. Instrument: TYXN-91 Intelligent Platelet Aggregation Instrument (produced by Shanghai General Electromechanical Technology Institute)

Test method

Rats were randomly divided into 11 groups, namely: solvent control group, ASA 100mg / kg, N, N,-dibenzylethylenediamine salt 200mg / kg, 100mg / kg, 50mg / kg group, bell salt 200mg / kg , 100mg / kg, 50mg / kg group, calcium salt 200mg / kg, 100mg / kg, 50mg / kg group. Oral administration was performed once a day for three consecutive days, and the solvent control group was given the same amount of solvent. Blood was collected from the common carotid artery 1 hour after the last dose, and sodium citrate (3.8%) was mixed with whole blood at a ratio of 1: 9 for anticoagulation, and platelet-rich plasma (PRP) and platelet-poor plasma ( PPP), adjust the PRP with PPP to a platelet number of 6-7xl O ^u L- Refer to Born's turbidimetry (GVR Born and MJ CROSS: The Aggregation of Blood Plate let s, J. Phys iology, 1963, 168: 178 ), Take 200μ1 of the adjusted PRP into the turbidimetric tube, and incubate at 37 ° C for 5 minutes, then add the inducer ADP to a final concentration of 5μηιο ¹ L " ¹ , determine the addition Maximum platelet aggregation rate and aggregation inhibition rate within 5 minutes after ADP.

 Aggregation inhibition rate = (aggregation rate of the solvent control group-aggregation rate of the administration group) / aggregation rate of the solvent control group χ 100 ° /.

 V. Test results

 The results showed that the platelet aggregation rate of rats in the N, N, -dibenzylethylenediamine salt 50-200mg / kg group, potassium salt and calcium salt 100 mg / kg, and 200 mg / kg group was similar to that of the solvent control group. There were significant differences in the ratios, and there was no significant difference in the platelet aggregation rate between the potassium and calcium 50 mg / kg rats and the solvent control group. At the same dose, the inhibitory effect of N, N, -dibenzylethylenediamine salt on platelet aggregation in rats was stronger than that of potassium and calcium salts. The results are shown in Table 6.

 Effect on ADP-induced platelet aggregation in rats

 Note: Compared with the solvent control group, *** P <0. 001, ** P <0. 01, * P <0. 05 Example 7: Preparation of tablets

 Table 7 Tablet formulations

 Ingredient quantity (mg / tablet)

 N, N, -Dibenzylethylenediamine salt 200

 Starch 50

 Microcrystalline cellulose 35

 Magnesium stearate 0.5

 Talcum powder 1

 Carboxamidine sodium 5

Preparation method: According to the formula in Table 7, N, N, -dibenzylethylenediamine salt, starch, micro The crystalline cellulose and sodium carboxymethylcellulose are mixed uniformly, wetted with water uniformly, granulated, dried, and whole granulated, magnesium stearate and talc are added, and the mixture is compressed to obtain tablets of this product.

: Preparation of stocks

 Table 8 Capsule formula

 Ingredients Quantity (mg / capsule)

 N, N '-Dibenzylethylenediamine salt 200

 Starch 50

 Methyl cellulose 3

 Cross-linked PVP 1

 Preparation method: According to the formula in Table 8, N, N, -dibenzylethylenediamine salt and excipients are mixed uniformly, granules are made by wet method, dried and whole granules, and then the mixture is charged into gastric hard gelatin capsules in a certain amount, that is, Get this product.

: Preparation of granules

 Table 9 Granule formulations

 Ingredient Quantity (mg / pack)

 N, N, -Dibenzylethylenediamine salt 200

 Starch 1000

 Dextrin 150

 Powdered sugar 150

 Preparation method: According to the formula in Table 9, N, N, -dibenzylethylenediamine salt and auxiliary materials are mixed and then wet granulated, dried, whole and classified, and divided into packages to obtain the granules of this product.

Claims

Rights request  1.2-(α-Hydroxypentyl) phenylarsinic acid Ν, Ν, -dibenzylethylenediamine salt The structure is as follows:

2. A method for preparing 2- (oc-hydroxypentyl) benzoic acid N, N'-dibenzylethylenediamine salt according to claim 1, comprising the following steps:  (1) Perform acidification operation on 2-(α-hydroxypentyl) phenylarsonate to control the temperature of the acidification process -10 ~ 10 ° C; pH 2-3, then extract with an organic solvent, and wash the organic solvent layer to medium After drying, the organic solvent was removed under reduced pressure to obtain 2-(oc-hydroxypentyl) phenylarsinic acid;  (2) The 2- (α-hydroxypentyl) phenylarsinic acid in step (1) is dissolved in an organic solvent, and a solution prepared by adding N, N, -dibenzylethylenediamine in an organic solvent is reacted to control, The reaction temperature is -10 ~ 10 ° C;  (3) The reaction product in step (1) is stirred for several hours and then filtered, the filter cake is washed with an organic solvent, and the filter cake is dried to obtain 2- (oc-hydroxypentyl) phenylarsinic acid N, N, -di Benzylethylenediamine salt.  3. The method according to claim 2, wherein the organic solvent is diethyl ether, dichloromethane or ethyl acetate.  4. The method according to claim 2, wherein the organic solvent is diethyl ether.  5. The preparation method according to any one of claims 2-4, wherein the reaction temperature in step (2) is 0 ~ -5 ° C. 6. The use of the salt according to claim 1 in the preparation of a medicament for preventing and / or treating cardio-cerebral ischemia and / or cardio-cerebral artery occlusion and improving cerebral circulation. 7. A pharmaceutical composition for preventing and / or treating cardio-cerebral ischemia and / or cardio-cerebral artery occlusion and improving cerebral microcirculation, characterized in that it contains a therapeutically effective amount of the salt according to claim 1.  8. The pharmaceutical composition according to claim 7, characterized in that it is made into a tablet, capsule or granule preparation.