Compounds for treating cardiovascular and cerebrovascular diseases and preparation methods thereof

Abstract

The present invention provides a compound for treating cardiovascular and cerebrovascular diseases and a preparation method thereof. The compound prepared by the present invention has better drug properties in terms of physical and chemical properties. In an ischemic myocardial infarction model, the compound of the present invention can significantly reduce the myocardial infarction area, and the effect is better than that of a positive drug control group, indicating that it can be used for the prevention and treatment of ischemic cardiovascular and cerebrovascular diseases.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a compound for treating cardiovascular and cerebrovascular diseases and a preparation method thereof. Background Art

[0002] Cardiovascular and cerebrovascular diseases are serious threats to human health, and are especially common among the elderly. They are characterized by high morbidity, high disability and high mortality. Even with the most advanced and complete treatment methods currently available, more than 50% of survivors of cardiovascular and cerebrovascular accidents are unable to take care of themselves completely, and most of them require lifelong medication.

[0003] As my country gradually enters an aging society, cardiovascular and cerebrovascular diseases are bringing an increasingly heavy burden to society and families, so it is of great significance to research and develop new and efficient drugs for the prevention and treatment of cardiovascular and cerebrovascular diseases. Summary of the invention

[0004] The present invention provides a compound for treating cardiovascular and cerebrovascular diseases and a preparation method thereof. The present invention prepares a new compound that is safer, more effective, and has controllable quality, and the compound has low difficulty in preparation and is more suitable for clinical application, and can meet people's demand for the continuous improvement of drug quality. The method of the present invention has a simple reaction system, and the product is easy to separate and purify. By controlling the reaction conditions, the reaction efficiency, the purity and yield of the product can be improved, and it has broad application prospects. At the same time, the present invention has studied the pharmacological effects of the prepared new compound and found that it has significant therapeutic effects in the prevention and treatment of ischemic cardiovascular and cerebrovascular diseases.

[0005] In a first aspect of the present invention, there is provided a compound represented by formula I or a salt, optical isomer, hydrate, or solvate thereof, wherein the structure of the compound is as follows:

[0006]

[0007] The second aspect of the present invention provides a pharmaceutical composition, which comprises the compound described in the first aspect of the present invention or its salt, hydrate, or solvate, and a pharmaceutically acceptable carrier or excipient.

[0008] In another preferred embodiment, the pharmaceutical composition is in the form of an oral preparation and / or an injection preparation.

[0009] The third aspect of the present invention provides the use of the compound or its salt, hydrate, or solvate described in the first aspect of the present invention, or the pharmaceutical composition described in the second aspect of the present invention, for preparing a drug for preventing and / or treating cardiovascular and cerebrovascular diseases.

[0010] In another preferred embodiment, the cardiovascular and cerebrovascular diseases include ischemic cardiovascular and cerebrovascular diseases and their complications, preferably including myocardial ischemia, myocardial hypoxia, myocardial infarction, stroke (such as ischemic stroke), cerebral infarction or cerebral arteriosclerosis.

[0011] In another preferred embodiment, the dosage form of the drug is an oral preparation and / or an injection preparation.

[0012] The fourth aspect of the present invention provides a method for preparing the compound described in the first aspect of the present invention, comprising the steps of:

[0013]

[0014] In an inert solvent, compound II and compound III are reacted to obtain a compound of formula I.

[0015] In another preferred embodiment, the reaction is carried out under the catalysis of a catalyst, and the catalyst is a weakly alkaline catalyst, preferably selected from the following group: sodium bicarbonate, sodium carbonate, magnesium carbonate, magnesium hydroxide, calcium hydroxide, or a combination thereof.

[0016] In another preferred embodiment, the inert solvent is DMF.

[0017] In another preferred embodiment, the ratio of compound II to compound III is 1:(0.2-5), preferably 1:(0.5-3), and more preferably 1:(1-2).

[0018] In another preferred embodiment, the reaction is carried out at a temperature T, which is 10-40°C, preferably 20-30°C, and more preferably 25°C.

[0019] In another preferred embodiment, the reaction is carried out under an inert gas.

[0020] In another preferred embodiment, the steps include: dissolving compound II in an inert solvent (such as DMF), adding a catalyst and cooling to a first temperature T 0 , add compound III in an inert gas and react at temperature T to obtain a compound of formula I.

[0021] In another preferred embodiment, the first temperature T 0 0-4℃.

[0022] In another preferred embodiment, the reaction time is 3-48 h, preferably 12-24 h.

[0023] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0024] The following will further describe the concept, specific structure and technical effects of the present invention to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The synthetic route diagram of the present invention and the structure of the product.

[0026] Figure 2 HPLC spectra and thin layer chromatograms of the raw materials and products of the present invention.

[0027] Figure 3 The HPLC spectrum of the product obtained by purifying the product of the present invention through preparative HPLC.

[0028] Figure 4 Mass spectrum of the product of the present invention.

[0029] Figure 5 The NMR hydrogen spectrum of the product of the present invention.

[0030] Figure 6 The NMR carbon spectrum of the product of the present invention.

[0031] Figure 7 IR spectrum of the product of the present invention.

[0032] Figure 8 UV spectrum of the product of the present invention.

[0033] Fig. 9 The product of the present invention resists H under different concentration conditions 2 O 2 Induced apoptosis effect of H9C2 cardiomyocytes. DETAILED DESCRIPTION

[0034] The present inventors have developed a new compound (Compound I, (E)-((R)-3-(3,4-dihydroxyphenyl)-1-oxo-1-((3,5,6-trimethylpyrazin-2-yl)methoxy)pro pan-2-yl)3-(2-((E)-3,4-dihydroxystyryl)-3,4-dihydroxyphenyl)acrylate) through extensive and in-depth research, and disclosed its preparation method and application. The synthesis route of the new compound is shown in Figure 1. After the product was purified by preparative HPLC, the structure of the compound was determined by nuclear magnetic resonance, mass spectrometry, infrared, ultraviolet and other methods. Compared with its structural unit compounds II and III, the compound I of the present invention is more drugable in terms of physical and chemical properties. In the ischemic myocardial infarction model, the compound I of the present invention can significantly reduce the area of ​​myocardial infarction, and the effect is better than that of the positive drug control group, compound II group and compound III group, indicating that it can be used in the prevention and treatment of ischemic cardiovascular and cerebrovascular diseases, and has obvious advantages.

[0035] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0036] When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, -CH2O- is equivalent to -OCH2-.

[0037] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0038] As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0039] The salts that may be formed by the compounds of the present invention also belong to the scope of the present invention. Unless otherwise specified, the compounds of the present invention are understood to include their salts. The term "salt" used herein refers to an acidic or basic salt formed with an inorganic or organic acid and a base. In addition, when the compound of the present invention contains a basic fragment, it includes but is not limited to pyridine or imidazole, and contains an acidic fragment, including but not limited to carboxylic acid, the zwitterions ("inner salts") that may be formed are included in the scope of the term "salt". Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, in separation or purification steps in the preparation process. The compounds of the present invention may form salts, for example, the compound of formula I reacts with a certain amount of acid or base, such as an equivalent amount, and is salted out in a medium, or freeze-dried in an aqueous solution.

[0040] The compounds of the present invention contain basic fragments, including but not limited to amines or pyridine or imidazole rings, which may form salts with organic or inorganic acids. Typical acids that can form salts include acetates (such as acetic acid or trihaloacetic acid, such as trifluoroacetic acid), adipates, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphor salt, camphorsulfonate, cyclopentanepropionate, diglycolate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide.

[0013] The invention also includes but is not limited to the following: esters, isothioates (e.g., 2-hydroxyethanesulfonate), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonate), nicotinates, nitrates, oxalates, pectinates, persulfates, phenylpropionates (e.g., 3-phenylpropionate), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., formed with sulfuric acid), sulfonates, tartrates, thiocyanates, toluenesulfonates such as p-toluenesulfonates, dodecanoates, and the like.

[0041] The acidic fragments that the compounds of the present invention may contain, including but not limited to carboxylic acids, may form salts with various organic or inorganic bases. Typical base-formed salts include ammonium salts, alkali metal salts such as sodium, lithium, potassium salts, alkaline earth metal salts such as calcium, magnesium salts, and salts formed with organic bases (such as organic amines), such as benzyl, dicyclohexylamine, hepamine (salts formed with N,N-di(dehydroabietyl)ethylenediamine), N-methyl-D-glucamine, N-methyl-D-glucamide, tert-butylamine, and salts formed with amino acids such as arginine, lysine, etc. Basic nitrogen-containing groups can be combined with halide quaternary ammonium salts, such as small molecule alkyl halides (such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl), dialkyl sulfates (such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and dipentyl sulfate), long chain halides (such as chlorides, bromides and iodides of decyl, dodecyl, tetradecyl and tetradecyl), aralkyl halides (such as benzyl and phenyl bromides), etc.

[0042] The compounds of the present invention are understood to include pharmaceutically acceptable derivatives thereof. The term "pharmaceutically acceptable derivative" may include any pharmaceutically acceptable salt, hydrate or prodrug or any other compound, or when administered to a subject, can provide (directly or indirectly) a compound of formula I, or an active metabolite or residue thereof.

[0043] Prodrugs and solvates of the compounds of the present invention are also within the scope of the invention. The term "prodrug" herein refers to a compound that undergoes chemical transformation by metabolic or chemical processes to produce a compound, salt, or solvate of the present invention when treating a related disease. "Solvate" refers to a compound formed by the interaction of a compound of the present invention with a solvent.

[0044] The compounds, salts or solvates of the present invention may exist in tautomeric forms (such as amides and imino ethers). All these tautomers are part of the present invention.

[0045] Unless otherwise specified, the structural formulas described in the present invention are intended to include all isomeric forms (such as enantiomers, diastereomers and geometric isomers (or conformational isomers)): for example, R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers or geometric isomers (or conformational isomers) are all within the scope of the present invention.

[0046] All stereoisomers of the compounds (e.g., those due to asymmetric carbon atoms that may exist for various substitutions), including enantiomeric and diastereomeric forms, are contemplated by the present invention. Individual stereoisomers of the compounds of the present invention may not exist simultaneously with other isomers (e.g., as a pure or substantially pure optical isomer having a particular activity), or may be mixtures, such as racemates, or mixtures with all other stereoisomers or portions thereof. The chiral centers of the present invention have two configurations, S or R, as defined by the 1974 recommendations of the International Union of Pure and Applied Chemistry (IUPAC). Racemic forms can be resolved by physical methods, such as fractional crystallization, or by crystallization of diastereoisomers derived therefrom, or by separation by chiral column chromatography. Individual optical isomers can be obtained from racemates by suitable methods, including but not limited to conventional methods, such as recrystallization after salt formation with an optically active acid.

[0047] The compounds of the present invention are prepared, separated and purified to obtain compounds with a weight content of equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 99% ("very pure" compounds), which are listed in the text description. Such "very pure" compounds of the present invention are also considered part of the present invention.

[0048] Throughout the specification, groups and substituents may be selected to provide stable fragments and compounds.

[0049] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0050] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intraperitoneal administration, intramuscular administration or subcutaneous administration).

[0051] In a specific embodiment, the steps for preparing the compound of the present invention are as follows: Compound II is dissolved in 40 mL of dry DMF, and then NaHCO is added. 3 , stirred evenly at room temperature, the reaction system was cooled to 0 °C, N 2 Under protection, compound III was added and the reaction was carried out at 25°C for 12 hours.

[0052] The present invention is further described below in conjunction with specific implementation cases. The following examples are only used to illustrate the present invention, but are not used to limit the scope of the present invention. Therefore, the scope of protection of the patent of the present invention shall be subject to the attached claims. The experimental materials and reagents used in the following examples can be obtained from commercial channels unless otherwise specified. Normal temperature or room temperature refers to 4°C-25°C, preferably 15-25°C.

[0053] Example 1 Synthesis of Compound I of the Present Invention

[0054] The synthetic raw material intermediates compound II (CAS No.: 96574-01-5) and compound III (CAS No.: 79074-45-6) used in this example can be obtained from commercial channels, and the HPLC purity of compound II and compound III used in this example is about 97%. Other reagents involved are all common analytical grade reagents.

[0055] Take a round-bottom flask, add 11.45 g of compound II powder, and add 40 mL of DMF (dimethylformamide) to dissolve it. Then add NaHCO 3 (2.5 g), stirred at room temperature. Then the reaction system was cooled to 0 °C, N 2Under protection, add 5g of compound III and react at 25°C for 12 hours. The reaction equation is as follows: Figure 1 shown.

[0056] After the reaction is completed, TLC is used to detect the reaction situation, and the developing solvent is toluene: chloroform: ethyl acetate: formic acid (1:1.5:2:1). 40 mL of ice brine is added to the reaction system for dilution, and then ethyl acetate is used for extraction (150 mL*5). The ethyl acetate layers are combined and washed three times with an equal volume of ice brine to remove DMF. The ethyl acetate layer is detected by HPLC.

[0057] HPLC spectrum and TLC test results are as follows Figure 2 From TLC, it can be seen that compound II still remains, compound III is basically reacted completely, and new products are produced. From the HPLC spectrum, it can be seen that the compound with a retention time of 16 minutes is the product.

[0058] The product was purified by preparative HPLC, and its structure was confirmed by mass spectrometry, nuclear magnetic resonance and other related methods as shown in Formula I ((E)-((R)-3-(3,4-dihydroxyphenyl)-1-oxo-1-((3,5,6-trimethylpyrazin-2-yl)methoxy)propan-2-yl)3-(2-((E)-3,4-dihydroxystyryl)-3,4-dihydroxyphenyl)acrylate) Figure 4-Figure 8 ). Figure 4 Mass spectrum of the product of the present invention. Figure 5 The NMR hydrogen spectrum of the product of the present invention. Figure 6 The NMR carbon spectrum of the product of the present invention. Figure 7 IR spectrum of the product of the present invention. Figure 8 UV spectrum of the product of the present invention.

[0059] The purity of compound I obtained by preparative HPLC purification in this example is 95% (HPLC spectrum as shown in Figure 3 ). The final amount of the product obtained in this example is 10.3 g, and the reaction yield is 70.2% calculated based on compound III.

[0060] Example 2 Effect of Catalyst on Reaction

[0061] In this example, a method similar to that of Example 1 is used, except that the catalysts are different, and the effects of different catalysts on the reaction yield are studied. The weakly alkaline catalysts used in this example include sodium bicarbonate (same as in Example 1), sodium carbonate, magnesium carbonate, magnesium hydroxide, and calcium hydroxide, and the amount of addition is the same as the amount of sodium bicarbonate in Example 1 (30 mmol). The remaining reaction conditions are the same as in Example 1, and the compound obtained in Example 1 is finally calculated as a reference substance. The amount of the product finally obtained by different catalysts is as shown in Table 1 below:

[0062] Table 1 Effect of catalyst type on product amount and reaction yield

[0063]

[0064] Based on the results of this example, it can be found that different weakly alkaline catalysts have certain differences in the catalytic effects on the reaction, but all can complete the catalytic reaction. Finally, the weakly alkaline catalyst studied in this example is selected as the catalyst to be selected.

[0065] Example 3 Study on the ratio of reaction raw materials

[0066] The reaction raw materials used in the present invention are mainly compound II and compound III. Considering the cost of raw materials, it is chosen to increase the amount of relatively low-cost raw materials to promote the complete reaction of high-cost raw materials as much as possible. To this end, it is necessary to study under what degree of raw material ratio conditions, the reaction results are still acceptable.

[0067] In this embodiment, the method of Example 1 is referred to, except that the amounts of the reaction raw materials Compound II and Compound III are adjusted (when the amount of Compound II is changed, Compound III is maintained at 5.0 g, and when the amount of Compound III is changed, the amount of Compound II is maintained at 11.45 g), and the yield of the reaction under different ratio conditions is studied. The final results are shown in Table 2:

[0068] Table 2 Effect of reaction raw material ratio on reaction yield

[0069]

[0070]

[0071] As shown in the table, when the reaction material ratio is close to 1: 1, the yield is the lowest. When one of the materials is increased, the reaction is promoted to the right, and the yield is significantly improved with less raw materials. However, blindly increasing a certain material will also cause more raw materials to react incompletely, thereby causing waste. Based on the results of the present embodiment, the final selection of the amount ratio of compound II to compound III is 1: 0.2-5. From the perspective of sufficient reaction of the raw materials, 1: 1-2 is more preferred.

[0072] Example 4 Reaction Temperature Study

[0073] This example uses the same test method as in Example 1, except that the reaction temperature is adjusted, and real-time sampling is performed during the reaction to monitor the maximum reaction yield and the time required to reach the corresponding yield under different reaction temperature conditions, and finally determine the appropriate reaction temperature. The detailed results are shown in Table 3 below:

[0074] Table 3 Effect of reaction temperature on reaction time and reaction yield

[0075]

[0076] As can be seen from the table above, with the increase of reaction temperature, the reaction rate increased significantly, but the by-products of the reaction also continued to increase, resulting in a low yield under the premise that there was no residual raw material. The reaction rate at low temperature was significantly reduced. After 48 hours of reaction at 10°C, although some raw materials still remained in the system, the yield increased very slowly. Continuing the reaction may slightly increase the yield, but the efficiency is low, and the reaction was finally terminated at 48 hours.

[0077] Based on the results of this example, the reaction temperature was finally selected to be 10-40°C, and the optimal reaction temperature was 25°C. The reaction time was adjusted in time according to the monitoring of the reaction yield under the corresponding temperature conditions. Example 5 MTT method for determining the effect of positive drugs and compound I of the present invention on H 2 O 2 Inhibitory effect on cardiomyocyte apoptosis caused by

[0078] Rat myocardial cells H9C2 (available from Shanghai Yaji Biotechnology Co., Ltd.) were selected to study the effects of positive drugs and compound I of the present invention on H 2 O 2 Inhibitory effect on cardiomyocyte apoptosis.

[0079] H9C2 cells were cultured as described in the preparation and culture instructions, counted, and inoculated in a 96-well plate at 3,000 cells / well, and prepared for the experiment after 24 hours.

[0080] In this experiment, there were two groups: an untreated group (no treatment, normal culture), a negative control group (addition of H 2 O 2 treatment without adding drugs), positive control group (addition of H 2 O 2 treatment, adding positive drug Danshen Chuanxiongqin injection (available from Jilin Sichang Pharmaceutical Co., Ltd.), the concentration is 200 mg / L according to 2,3,5,6-tetramethylpyrazine), the sample group to be tested (adding H 2 O 2treatment, adding the compound I of the present invention prepared in Example 1, and finally maintaining the samples to be tested at different concentrations).

[0081] After the cells were cultured in 96-well plates for 24 h, the sample solutions with different treatments were added to the culture medium. After further culture for 8 h, H 2 O 2 Treat with 1.0 mM of DMSO. After 2 hours, discard the supernatant of the culture medium and replace it with new culture medium. After 24 hours, add 20 μL of MTT to each well and continue to culture for 4 hours. Discard the supernatant, add 150 μL of DMSO to each well, and shake it slightly to dissolve. Read the absorbance at 570 nm to represent the cell viability. The untreated group is taken as the viability value of 100%, and the other groups are compared with it to calculate the relative cell viability. The results are shown in Fig. 9 shown.

[0082] The results showed that after H 2 O 2 The viability of the treated H9C2 cells was significantly affected, and the positive drug Danshen Chuanxiongqin injection had a certain protective effect on myocardial cells at a dose of 200 mg / L, and the effect was significantly different from that of the negative control group. More obviously, the compound I of the present invention (test sample group) showed a good protective effect when the dose reached 10 mg / L, and when the dose reached 50 mg / L, its effect was not only significantly better than the negative control group, but also significantly better than the positive drug control group, and when the dose was larger, the difference was more significant. The above results show that the compound I of the present invention can significantly protect the oxidative damage of myocardial cells at the cellular level, and the effect is significantly better than the negative control group and the positive drug group.

[0083] Example 6 Protective effect of compound I of the present invention on the heart of rats with myocardial ischemia

[0084] In this example, SD rats were used as experimental animals to construct a rat myocardial ischemia model, and then the protective effect of the drug on the heart of rats with myocardial ischemia was determined by oral administration and injection.

[0085] The reagents used in this example were prepared by the method in Example 1 (or the same source as in Example 1), and the remaining reagents were all commercially available common reagents. The experimental method used is as follows: each group of animals was continuously administered for one week, and the drug was administered once a day according to the set dose. The model was established 1 hour after the last administration, and the rat heart ischemia model was established by coronary artery ligation. 2 ml of blood was collected from the femoral artery of the rat 4 hours after modeling, and serum was retained for the detection of five myocardial enzymes (CK, CK-MB, LDH, ALT and AST). Then the rat was killed, the heart was removed, rinsed with physiological saline, and quickly frozen in a -20°C refrigerator for 10 minutes, and then the heart was equally divided into 5 pieces of myocardial slices of uniform thickness, and stained in 37°C 0.5% TTC dye solution for about 5 minutes. After TTC staining, the infarcted myocardium was white and the non-infarcted myocardium was red. The white part of the myocardium in the infarcted area was trimmed and weighed. The ratio of the myocardial weight in the infarcted area to the whole heart weight was calculated. The results are shown in Tables 4 and 5 below (respectively, the relevant data after intraperitoneal injection and oral administration):

[0086] Table 4 Protective effects of each component on myocardial ischemia model animals after intraperitoneal injection (mean±SD, n=10)

[0087]

[0088] Table 5 Protective effects of each component on myocardial ischemia model animals after oral administration (mean±SD, n=10)

[0089]

[0090]

[0091] ip: intraperitoneal injection; ig: oral administration; the same “sham operation group” and “model control group” were used for both oral and injection experiments (because neither of them required medication).

[0092] In this example, the protective effects of compound I of the present invention and its main structural unit compound II, 2,3,5,6-tetramethylpyrazine on myocardial ischemia in rats were studied by injection and oral administration, wherein the model control group was not administered any drug, and the sham operation group was not subjected to coronary artery ligation.

[0093] The results showed that when administered by injection, all three could produce good protective effects, and the protective effect produced by the compound I of the present invention was significantly stronger, and in terms of myocardial infarction proportion and multiple indicators of myocardial enzymes, it was not only significantly better than the model control group, but also significantly better than the compound II and 2,3,5,6-tetramethylpyrazine groups. When administered orally, the compound II and 2,3,5,6-tetramethylpyrazine groups only showed very weak therapeutic effects, while the compound I of the present invention showed a good therapeutic effect.

[0094] In summary, the results of this example show that compound I of the present invention can significantly reduce the infarction proportion during myocardial ischemia and improve the adverse consequences of myocardial damage. The effect is significantly better than that of the model control group without medication, and is also significantly better than that of the compound II group and the 2,3,5,6-tetramethylpyrazine group.

[0095] Example 7 Protective effect of compound I of the present invention on cerebral ischemia in rats

[0096] In this example, SD rats were used to construct a rat cerebral ischemia model by middle artery embolization. The protective effects of the compound of the present invention and its main structural unit compound II, 2,3,5,6-tetramethylpyrazine on cerebral ischemia in rats were determined by oral administration and injection. The test method is rat middle artery embolization-electrocoagulation method, which is as follows:

[0097] The rats were given drugs orally or intraperitoneally 1 hour before modeling, and then the modeling was started. The rats were anesthetized with 15% chloral hydrate 300 mg / kg intraperitoneally, and fixed on the rat operating table in the left lateral position. The left temporal top and face were shaved, and the skin was cut between the left eye and the left ear after disinfection with 75% ethanol. The temporal muscle and masseter muscle were bluntly separated to expose the pterygoid plate of the temporal bone. Under the operating microscope, a 2mm×2mm bone window was ground with a skull drill at 1mm near the mouth of the temporal bone and temporal squamous bone, and the skull was pried open with a crowbar. At this time, a straight blood vessel with fewer branches can be seen through the dura mater, which is the middle cerebral artery. Bipolar electrocoagulation forceps were used to cauterize between 1mm in the olfactory tract and the inferior cerebral vein, and the blood flow was completely blocked and the bleeding was stopped. The temporal muscle and skin were sutured in turn. After the rats woke up, they were returned to the cage and continued to be raised for 24 hours. The rats were anesthetized again with 15% chloral hydrate 300 mg / kg intraperitoneally, and the brains were removed by decapitation, and the olfactory bulbs, cerebellum and brainstem were removed and quickly frozen in a -20°C refrigerator for about 20 minutes; the frozen brains were cut into 7 equal pieces; the brain slices were placed in 0.5% TTC staining solution for staining (incubated at 40°C in the dark for 10 minutes), and the infarcted areas of the stained brain slices were white, and the non-infarcted areas were rose red; after staining, the brain slices were moved to 10% formaldehyde solution, stored in the dark for 24 hours, and finally photographed with a digital camera, and the areas of the front and back infarcted areas and non-infarcted areas were measured using image analysis software (ImageJ, version: 1.4.3.67), and the percentage of the infarcted area to the total area of ​​the infarcted cerebral hemisphere and the percentage of the corresponding treatment to reduce the infarcted area were calculated.

[0098] Table 6 Effects of each component on cerebral infarction area in cerebral ischemia model animals (mean ± SD, n = 10)

[0099]

[0100]

[0101] ip: intraperitoneal injection; ig: oral administration.

[0102] This example adopts two administration modes, injection and oral administration, to study the effects of the compound I of the present invention and its main structural unit intermediate compound II and 2,3,5,6-tetramethylpyrazine on the infarction area of ​​rats with cerebral infarction. The results show that when administered by injection, they all have good activity in reducing the infarction area, among which the compound I of the present invention has a better effect, not only better than the model control group without medication, but also significantly better than the high-dose groups of the latter two.

[0103] In the oral administration experiment, the intermediate compound II and 2,3,5,6-tetramethylpyrazine had poor effects, while the compound of the present invention still had a good effect of reducing the area of ​​cerebral infarction after oral administration, and had higher oral bioavailability and efficacy, which were significantly better than its structural unit intermediate compound II and 2,3,5,6-tetramethylpyrazine.

[0104] In summary, the compound I of the present invention has significant therapeutic effect in reducing the cerebral infarction area in cerebral ischemia model animals and has good application prospects.

[0105] The preferred specific embodiments of the present invention are described in detail above. It should be understood that ordinary technicians in the field can make many modifications and changes based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by technicians in the field of technology through logical analysis, reasoning or limited experiments based on the concept of the present invention on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. A compound represented by formula I or a salt thereof, It is characterized in that The structure of the compound is as follows:

2. A pharmaceutical composition, It is characterized in that The pharmaceutical composition comprises the compound or salt thereof according to claim 1, and a pharmaceutically acceptable carrier or excipient.

3. The pharmaceutical composition according to claim 2, It is characterized in that The dosage form of the pharmaceutical composition is an oral preparation and / or an injection preparation.

4. Use of the compound or salt thereof according to claim 1, or the pharmaceutical composition according to claim 2, It is characterized in that Used for preparing medicines for preventing and / or treating cardiovascular and cerebrovascular diseases, wherein the cardiovascular and cerebrovascular diseases are myocardial ischemia or ischemic stroke.

5. The use according to claim 4, It is characterized in that The cardiovascular and cerebrovascular disease is ischemic stroke.

6. A method for preparing the compound or a salt thereof according to claim 1, It is characterized in that Includes steps: In an inert solvent, compound II and compound III react to obtain compound I, which is a compound represented by formula I; The reaction is carried out under the catalysis of a catalyst, and the catalyst is a weakly alkaline catalyst; The molar ratio of compound II to compound III is 1:(0.2-5); The reaction is carried out at a temperature T, which is 10-40°C.

7. The preparation method according to claim 6, It is characterized in that The catalyst is selected from the group consisting of sodium bicarbonate, sodium carbonate, magnesium carbonate, magnesium hydroxide, calcium hydroxide, or a combination thereof.

8. The preparation method according to claim 6, It is characterized in that The inert solvent is DMF.

9. The preparation method according to claim 6, It is characterized in that The molar ratio of the compound II to the compound III is 1:(0.5-3).

10. The preparation method according to claim 9, It is characterized in that The molar ratio of the compound II to the compound III is 1:(1-2).

11. The preparation method according to claim 6, It is characterized in that The temperature T is 20-30°C.

12. The preparation method according to claim 11, It is characterized in that The temperature T is 25°C.

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