CN112209920A

CN112209920A - Oxygen-containing heterocyclic compound for treating cerebral apoplexy

Info

Publication number: CN112209920A
Application number: CN202011301356.0A
Authority: CN
Inventors: 任力杰; 谭回; 胡诗雨; 冯宏业; 王洋; 雷志浩
Original assignee: Shenzhen Second Peoples Hospital
Current assignee: Shenzhen Second Peoples Hospital
Priority date: 2020-11-19
Filing date: 2020-11-19
Publication date: 2021-01-12
Anticipated expiration: 2040-11-19
Also published as: CN112209920B

Abstract

The invention relates to a compound for preventing and treating cerebral apoplexy, in particular to ischemic cerebral apoplexy, which shows good therapeutic activity in a mouse cerebral ischemia reperfusion test, can obviously reduce the proportion of cerebral infarction and can be used for preventing and treating cerebral apoplexy, in particular to ischemic cerebral apoplexy.

Description

Oxygen-containing heterocyclic compound for treating cerebral apoplexy

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a compound for preventing and treating cerebral apoplexy, especially ischemic cerebral apoplexy, and also relates to a preparation method, a medicinal composition and application of the compound.

Background

In recent years, the incidence rate of stroke ranks first in the world in China, and the third national cause of death investigation result proves that the stroke becomes the first cause of death in China, the annual growth rate exceeds 8.7%, and the life health of the people in China is seriously influenced. The stroke IS mainly divided into Ischemic Stroke (IS) and Hemorrhagic Stroke (HS), wherein the IS type patients account for three quarters of the total disease patients and are the main type of stroke disease. The key of IS treatment IS 'early diagnosis and early intervention', various current treatments try to save ischemic brain tissues by recanalizing blood flow of early infarcted brain tissues, but the short 'treatment time window' hinders the popularization of the treatment mode. For many patients, particularly in poor and basic areas with relatively backward medical conditions and relatively weak stroke treatment systems, the patients often exceed a treatment time window when hospitalizing, and the existing thrombolytic therapy is far from achieving the purpose of effective treatment. Therefore, an efficient and convenient-to-popularize IS treatment method IS urgently sought.

The existing medicine for treating cerebral apoplexy aims at blocking the neuronal necrosis caused by ischemia, prolonging the tolerant ischemic time and the treatment time window, enhancing the survival capability of neurons, reversing the penumbra, reducing the infarct volume and promoting the recovery of nerve function.

Edaravone is a novel neuroprotective agent developed by tokyo corporation of mitsubishi japan, is a potent radical scavenger, is marketed in japan in 2001, and is used for treating neuropathy caused by cerebral infarction, and has a good therapeutic effect on ischemic stroke in view of its antioxidant effect.

Coumarin is a lactone compound widely existing in nature, has various pharmacological activities such as oxidation resistance, blood sugar reduction, anticoagulation, antibiosis and the like on a human body, and recently researches show that coumarin and derivatives thereof also have physiological activities such as neuroprotection and the like, and can be used for cerebral nerve diseases such as cerebral apoplexy, cerebral ischemia and the like. Therefore, the inventor introduces edaravone and coumarin into the same molecular structure by the principle of molecular split, and screens out a compound with potential activity of treating cerebral apoplexy by a mouse cerebral ischemia-reperfusion test.

In summary, although various chemical drugs and natural drugs are in different research stages, no drug for splicing edaravone and coumarin fragments exists in the drugs for treating stroke on the market at present. Therefore, the development and research of new compounds with the activity of treating cerebral apoplexy have important clinical significance.

Disclosure of Invention

The technical problem to be solved by the invention is as follows:

in a first aspect of the present invention, there is provided a compound of formula I and pharmaceutically acceptable salts thereof, having the structure:

preferably, the pharmaceutically acceptable salt is selected from: hydrochloride, hydrobromide, phosphate, sulfate, acetate, oxalate, tartrate and the like;

in another aspect of the invention, there is provided a process for the preparation of a compound of formula I, the synthetic route for which is as follows:

the compound 2 and the compound 3 react under the action of a condensation reagent to generate the compound shown in the formula I.

The specific reaction steps are as follows: under the ice bath condition, adding the compound 2, Dicyclohexylcarbodiimide (DCC), 4-Dimethylaminopyridine (DMAP) and dichloromethane into a reaction bottle, stirring for 15-30min, adding a dichloromethane solution of the compound 3, heating to the reflux temperature, reacting for 5-10h, and performing post-treatment to obtain a target product I.

Preferably, the feeding molar ratio of the compound 2 to the compound 3 is 1:1-2, preferably 1: 1.1.

In another aspect of the present invention, a pharmaceutical composition is provided, which comprises a compound represented by formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

In another aspect, the invention relates to the use of a compound of formula I, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for the manufacture of a medicament for the treatment of stroke; preferably, the stroke is ischemic stroke.

Compared with the prior art, the invention has the beneficial effects that:

(1) the invention provides a new compound containing oxygen heterocyclic ring for treating cerebral apoplexy, which widens the range of the existing compound for treating cerebral apoplexy and can be continuously optimized as a lead compound;

(2) the compound shown in the formula I shows good therapeutic activity in a mouse cerebral ischemia reperfusion test, and can be used for preventing and treating cerebral apoplexy, particularly ischemic cerebral apoplexy.

Detailed Description

The present invention is described in more detail below to facilitate an understanding of the present invention. The compounds of the present invention may be prepared by a number of methods well known to those skilled in the art of synthesis. The compounds of formula I may be prepared using the reactions and techniques outlined below, together with methods known in the art of synthetic organic chemistry or variations thereof as understood by those skilled in the art. Preferred methods include, but are not limited to, those described below. The reaction is carried out in a solvent suitable for the reagents and materials used and for the conversion to be effected. Furthermore, in the synthetic methods described below, it is to be understood that all proposed reaction conditions (including choice of solvent, reaction atmosphere, reaction temperature, experimental duration and work-up procedures) are selected as standard conditions for the reaction, which should be readily ascertained by one skilled in the art of organic synthesis. Not all compounds falling within a given class may be compatible with certain reaction conditions required in certain of the described methods. These limitations on substituents compatible with reaction conditions will be apparent to those skilled in the art and alternative methods may be used.

Example 1: synthesis of Compound I

Under the ice bath condition, adding the compound 2(190mg, 1mmol), dicyclohexylcarbodiimide (413mg, 2mmol), 4-dimethylaminopyridine (244mg, 2mmol) and 10mL of dichloromethane into a 150mL three-neck flask, stirring for 20min, adding a dichloromethane (15mL) solution of the compound 3(209mg, 1.1mmol), heating to a reflux temperature after the addition is finished, reacting for 8h, cooling a reaction solution to room temperature after TLC detection reaction is finished, adding 30mL of water, standing for liquid separation, concentrating organic phase under reduced pressure, and performing silica gel column chromatography to obtain the target product I which is a white solid, wherein the yield is 41% and the HPLC purity is 98.1%.

The molecular formula is as follows: c₂₀H₁₄N₂O₅；ESI-MS：363.17[M+H]⁺

¹H NMR(400MHz，CDCl³)δ2.24(s，3H)，2.52(s，2H)，6.07(dd，1H)，7.22(d，2H)，7.54(d，1H)，7.71(m，1H)，7.96(m，2H)，8.58(m，2H)。

Example 2: application of compound shown as formula I in treating ischemic stroke

The pharmacological experiment of mouse line-tying method for preparing focal cerebral ischemia model is animal experiment for verifying that the medicine has the effect of preventing and treating ischemic cerebral apoplexy. An ICR mouse cerebral ischemia reperfusion model is prepared by a embolus method (the preparation method is detailed in the pharmacological and pharmacological toxicology part of the guidance and guidance principles of the preclinical research of new western medicines, the ministry of health of the people's republic of China, 1993: 73; pharmacological experimental methodology, second edition, the public health publishing agency, 1982: 830 and 1113), and the model mouse is used for testing in other groups except a pseudo-operation group.

1. Animal(s) production

ICR mice, male, weight 20 ~ 22 g. 20 per group, 3 groups in total, compound I group: 1 mg/kg; the sham operation group: giving physiological saline equal to the drug test group for false operation; model group: the cerebral ischemia reperfusion surgery was performed by administering physiological saline in an amount equivalent to that of the drug test group.

2. Experimental methods

Mice were anesthetized by intraperitoneal injection with 10% chloral hydrate, the neck was opened, and the proximal common carotid artery, the external carotid artery and the branch arteries were isolated and ligated. The right internal carotid artery was isolated, the palatine artery was isolated down the internal carotid artery, and the branch was ligated at the root. An artery clamp is arranged at the near end and the far end of an internal carotid artery, an incision is made at the bifurcation of a common carotid artery, a nylon wire is inserted, a plug wire enters the internal carotid artery and enters the skull to the anterior cerebral artery, and all blood flow sources of the middle cerebral artery are blocked. Removing the artery clamp, tightening the prepared thread, leaving a 1cm long thread end outside, and suturing the skin. The administration was intravenous 1 hour after ischemia. Reperfusion was performed after 1 hour of continued ischemia. Reperfusion for 8 hours and re-injection. The procedure was as above except that the sham group was not cannulated.

Mice surviving 24 hours after reperfusion were observed for the following indices:

(1) effect on cerebral infarct volume: 10 mice were taken from each group, brains were sacrificed and coronal brain slices approximately 2mm thick were excised and immediately placed in 2% TTC solution and incubated at 37 ℃ for 30 minutes. Infarcted areas appeared white and non-infarcted areas appeared red. The digital camera takes the record, measures the area of each area by computer image processing, and calculates the percentage (%) of the infarct area in the whole brain tissue.

(2) And observing the behavior change of the living mice 24 hours later, and performing behavioral scoring: reference Zea Longa's 5-score scoring criteria: 0 is normal and has no nerve injury symptom; 1 is divided into front paws which can not extend to the opposite side completely; 2, turning to the outside; 3, pouring towards the opposite side; 4, the patient cannot walk spontaneously and loses consciousness.

3. Results of the experiment

Table 1: experiment of cerebral infarction area after cerebral ischemia reperfusion of each group of mice

Note: in comparison to the set of models,^#P＜0.01

the test result shows that: the brain tissue of the mice in the sham operation group has no infarction. The ischemic lateral brain tissue of the model group mice has obvious infarction phenomenon, and the cerebral infarction/whole brain (%) is 50.6 +/-3.6%. The result proves that the test compound can play a good role in protecting cerebral infarction caused by the ischemia-reperfusion injury of the brain of a mouse.

Table 2: experimental groups of mice neurobehavioral Scoring after cerebral ischemia reperfusion

Grouping	Dosage (mg/Kg)	Behavioural scoring (score)
			Artificial operation group	-	0
Model set	-	3.8±0.4^#
			Compounds of group I	1	2.1±0.3^#

Note: in comparison to the set of models,^#P＜0.05

the test result shows that: the sham-operated mice did not show any abnormal symptoms. While model mice exhibited symptoms of nerve injury that either rolled laterally, or fell contralaterally, or failed to fully extend the contralateral forepaw. The behavior score of an ischemia-reperfusion mouse can be obviously reduced by the compound I at a lower dose, and the result proves that the test compound can obviously improve the nerve injury symptom caused by ischemia-reperfusion.

The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.

Claims

1. A compound of formula I and pharmaceutically acceptable salts thereof, having the structure:

2. a compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, selected from: hydrochloride, hydrobromide, phosphate, sulfate, acetate, oxalate, tartrate and the like.

3. A process for the preparation of a compound of formula I according to claim 1, which reaction scheme is as follows:

，

4. A pharmaceutical composition comprising a compound of formula I as described in any one of claims 1-2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound of formula I as defined in any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of stroke.

6. Use according to claim 5, wherein the stroke is ischemic stroke.