CN113150050B

CN113150050B - Novel ligustrazine derivative and preparation method and application thereof

Info

Publication number: CN113150050B
Application number: CN202011415504.1A
Authority: CN
Inventors: 郝杰杰; 王鹏; 李明; 白董慧; 于广利; 管华诗
Original assignee: Ocean University of China
Current assignee: Ocean University of China
Priority date: 2020-01-07
Filing date: 2020-12-04
Publication date: 2022-11-25
Anticipated expiration: 2040-12-04
Also published as: CN113150050A

Abstract

The invention provides a novel ligustrazine derivative and a preparation method and application thereof. Experiments prove that the ligustrazine derivative can effectively protect nerve cells and cardiac muscle cells from CoCl ₂ Hypoxic injury; and also has the function of obviously inhibiting the platelet aggregation. Therefore, the ligustrazine derivative can be used for preparing medicaments for effectively preventing or treating central nervous system diseases and/or thrombotic diseases; more importantly, the efficacy of the ligustrazine derivative is obviously superior to that of the ligustrazine with the same dosage; the invention widens the new application of the novel ligustrazine derivative and has great economic and social values.

Description

Novel ligustrazine derivative and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a novel ligustrazine derivative and a preparation method and application thereof.

Background

Cardiovascular and cerebrovascular diseases are diseases which seriously threaten human life and health in the modern society, and the clinical manifestations are cardiovascular and cerebrovascular ischemia or hemorrhage. According to the WHO estimation, 400 million people die of cardiovascular and cerebrovascular diseases every year in China by 2020. Stroke is also called cerebral stroke or cerebrovascular accident, the main clinical manifestations are cerebral ischemia and hemorrhagic injury, one thousand five million people suffer from stroke every year in the world, and in China, stroke is the first leading cause of human death and the third leading cause of disability. Most strokes are sudden death caused by oxidative stress in certain brain cells due to blood clots that cause the cerebral arteries to become blocked or ruptured. Ischemic stroke is a complex systemic disease with significant morbidity and mortality worldwide and requires significant medical and health expenditures, a serious problem in the medical world and even the society. Although many drugs have different degrees of curative effects on cerebral arterial thrombosis, the effect is not ideal, and at present, no drug of a certain class is effective on all patients with cerebral arterial thrombosis, namely, no specific drug in the true sense is provided for cerebral arterial thrombosis. Therefore, the development of a novel efficient medicament for treating cerebral arterial thrombosis has great practical significance. Currently, there are two main treatments for ischemic stroke: firstly, the blood oxygen supply of an ischemic region is increased by dissolving thrombus, and the clinical application of the blood oxygen supply agent mainly comprises vasodilators, anti-platelet aggregation drugs, anticoagulants and thrombolytic drugs; and protecting the neurons of ischemic tissues and preventing cells from excitotoxicity.

At present, because bioactive natural products have wide pharmacological actions, such as vascular endothelial cell protective activity, anticoagulation and antithrombotic activity, and the like, structural modification based on the natural products becomes an important strategy for treating ischemic stroke; the natural product is one of the important sources of the lead compounds of the innovative medicaments. Many drugs such as paclitaxel, artemisinin, etc. are derived from natural products. Due to its structural diversity and abundant stereochemical centers, natural active substances can be used as good leads for drug development. However, natural products do not necessarily meet the pharmaceutical requirements including physicochemical, biochemical, pharmacokinetic and safety properties of molecules, and require structural modification and optimization. The main directions of the structural modification of the existing natural products are as follows: improving biological activity, enhancing selectivity, improving physicochemical and biochemical properties, improving stability, improving pharmacokinetic properties, eliminating or reducing toxic and side effects and adverse reactions, etc.

Ligustrazine (ligustrazine) is separated from rhizoma Ligustici Chuanxiong, and is one of effective components of rhizoma Ligustici Chuanxiong. Ligustrazine has effects of inhibiting platelet aggregation, relieving angiospasm of smooth muscle, improving myocardial ischemia, and improving blood rheological property. However, pharmacokinetic studies find that ligustrazine has the disadvantages of fast metabolism, short half-life, low bioavailability and the like in animal bodies; clinical continuous administration is often performed to ensure that the concentration of ligustrazine in blood plasma can be kept at a certain level, but the toxicity of the drug is increased. Therefore, the research on the ligustrazine derivatives has attracted the wide attention of scientists. Lei Haimin in 2014 CN 201410587694 discloses the use of ligustrazine derivatives for protection of neuronal damage; zhu Yan equals 2017 and discloses a new use of ligustrazine stilbene derivatives in CN 201710058694; zhang Baohua equals 2019 and in CN 201910182300 discloses a nitrogen-containing mustard ligustrazine matrine derivative and its preparation method and application; however, there is no report of using ligustrazine in related diseases.

Disclosure of Invention

The invention aims to provide a novel ligustrazine derivative and a preparation method and application thereof. The series of novel ligustrazine derivatives can be used for preparing medicaments for preventing and treating diseases of nervous system, cardiovascular system and cerebrovascular system.

In order to realize the purpose of the invention, the invention adopts the following technical scheme to realize:

the invention provides a novel ligustrazine derivative, the structural formula of which is shown as the formula (I):

wherein R1 is any one of the following:

wherein R0= H or acetyl.

Further: the ligustrazine derivative is as follows:

wherein R = H or acetyl.

The invention also provides a preparation method of the ligustrazine derivative, which comprises the following steps:

(1) Using ligustrazine as a raw material, oxidizing with an oxidant to obtain a mononitrogen compound, heating with anhydride to generate Boekelheide rearrangement to obtain a compound which generates methyl acetate at the 2-position, and performing hydrolysis reaction under an alkaline condition to obtain 2-hydroxymethyl-3,5,6-trimethylpyrazine;

(2) Taking the 2-hydroxymethyl-3,5,6-trimethylpyrazine as a raw material, reacting with 1-bromo-2,3,4-acetyl-mannitol or 4-bromo-1,2,3-acetyl-mannitol and boron trifluoride diethyl ether, adding a saturated sodium bicarbonate solution, separating an organic phase, washing, drying, and performing column chromatography to obtain 2,3,4-acetyl mannitol-2-glucoside-3,5,6-trimethylpyrazine or 1,2,3-acetyl mannitol-2-glucoside-3,5,6-trimethylpyrazine; then removing protecting groups to obtain 2,3,4-hydroxy mannuronic acid-2-methoxy-3,5,6-trimethylpyrazine or 1,2,3-hydroxy mannuronic acid-2-methoxy-3,5,6-trimethylpyrazine;

2,3,4-hydroxy mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine or 1,2,3-hydroxy mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine can be acetylated under acetic anhydride/sulfuric acid condition to obtain 2,3,4-acetyl mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine or 1,2,3-acetyl mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine.

Further: the step (2) is washed by saturated sodium bicarbonate and water, and dried by anhydrous sodium sulfate.

The invention also provides the application of the ligustrazine derivative in preparing medicaments for preventing or treating diseases of a nervous system and a cardiovascular and cerebrovascular system.

Further: the nervous system diseases include Alzheimer's disease and Parkinson's disease.

Further: the cardiovascular and cerebrovascular system diseases comprise cerebral ischemic diseases, cerebral anoxia, myocardial anoxia, thrombosis resistance and blood circulation improvement.

The invention also provides a pharmaceutical composition containing the ligustrazine derivative and a pharmaceutically acceptable carrier.

Further: the pharmaceutical composition is formed into an oral preparation, specifically a tablet, an oral liquid, a pill, a capsule, a granule, an ointment, a drop pill, a syrup, a powder, a granule or a tincture.

Further: the pharmaceutical composition forms an injection, in particular to a powder injection or an injection.

The invention has the advantages and beneficial effects that:

1. the pathogenesis process of the diseases of the nervous system and the cardiovascular system is gradual and the mechanism is complex. The invention adopts CoCl ₂ The hypoxia-damaged myocardial and nerve cell damage model and the semi-body external platelet aggregation model are used for testing the protection effect of the ligustrazine derivative on nerve and myocardial cells and the effect of inhibiting platelet aggregation.

Experiments prove that the ligustrazine derivative can effectively protect nerves and myocardial cells from CoCl ₂ Platelet aggregation can also be inhibited by the effects of hypoxic injury. Therefore, the novel ligustrazine derivative can effectively prevent and treat diseases of nerve, cardiovascular and cerebrovascular systems through various ways.

2. More importantly, the effects of the novel ligustrazine derivative are obviously superior to those of the ligustrazine with the same dosage; the invention widens the new application of the novel ligustrazine derivative; can be developed into a novel medicine for preventing or treating diseases of the nervous system and the cardiovascular system, and has great economic and social values.

Detailed Description

The present invention will be further described with reference to the following examples, which are not intended to limit the scope of the present invention.

In the synthesis process of the series of compounds, because the number of the compounds is large and a plurality of synthesis steps are repeated, a representative synthesis route is selected for carrying out the embodiment statement:

example 1 preparation of series of Compounds A-D

In the preparation of the A series derivatives, 1-bromo-2,3,4 acetyl-manno ate and 4-bromo-1,2,3 acetyl-manno ate are first required, which can be prepared according to the preparation method well known to those skilled in the art, and there is no particular limitation to this application.

(1) Ligustrazine is used as a raw material, 30% hydrogen peroxide is used for oxidation at 70 ℃ to obtain a mononitrogen compound, then the mononitrogen compound and acid anhydride are subjected to Boekelheide rearrangement reaction under the heating condition to obtain a compound which generates methyl acetate at 2 position, and the compound and aqueous solution of 20 percent NaOH are subjected to hydrolysis reaction to finally obtain 2-hydroxymethyl-3,5,6-trimethyl pyrazine.

(2) Taking the 2-hydroxymethyl-3,5,6-trimethylpyrazine as a raw material, sequentially adding 1-bromo-2,3,4-acetyl-mannitol and boron trifluoride diethyl etherate in anhydrous dichloromethane under the protection of nitrogen, reacting at room temperature, adding a saturated sodium bicarbonate solution after the reaction is finished, separating an organic phase, washing with saturated sodium bicarbonate and water, drying with anhydrous sodium sulfate, performing column chromatography to obtain 2,3,4-acetyl-mannitol-2-glucoside-3,5,6-trimethylpyrazine, and supporting a protecting group in a methanol/sodium methoxide system to obtain 2,3,4-hydroxymannuronic acid-2-methoxy-3,5,6-trimethylpyrazine, namely a compound A0.

The compound A0 is acetylated under the condition of acetic anhydride/sulfuric acid by a conventional method to obtain 2,3,4-acetyl mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine, namely the compound A1.

(3) Taking 2-hydroxymethyl-3,5,6-trimethylpyrazine as a raw material, sequentially adding 4-bromo-1,2,3-acetyl-mannitol and boron trifluoride diethyl etherate in anhydrous dichloromethane under the protection of nitrogen, reacting at room temperature, after the reaction is finished, adding a saturated sodium bicarbonate solution, separating an organic phase, washing with saturated sodium bicarbonate and water, drying with anhydrous sodium sulfate, performing column chromatography to obtain 1,2,3-acetyl-mannitol-2-glucoside-3,5,6-trimethylpyrazine, and supporting a protecting group in a methanol/sodium methoxide system to obtain a compound C0, namely 1,2,3-hydroxymannuronic acid-2-methoxy-3,5,6-trimethylpyrazine.

The acetylation of C0 in conventional acetic anhydride/sulfuric acid process can obtain 1,2,3-acetyl mannuronic acid-2-methoxy-3,5,6-trimethyl pyrazine, compound C1.

Similarly, other compounds can be prepared according to the synthetic methods described in 1-2 and preparation methods well known to those skilled in the art.

The following compounds were prepared by the above synthetic and preparation methods:

A-1：

¹ H NMR(400MHz，DMSO-d ₆ )δ5.15(s，1H)，4.87(s，2H)，4.60(s，1H)，4.23(d，J＝5.0Hz，2H)，3.58(s，1H)，2.74(s，9H)，2.39(s，1H)，1.84(s，1H)，1.29(s，1H)。

¹³ C NMR(101MHz，DMSO-d ₆ )δ173.22(s)，149.76(s)，148.80(s)，147.40(s)，141.19(s)，101.65(s)，76.40(s)，74.56(s)，72.71(s)，71.72(s)，65.97(s)，22.34(d，J＝7.3Hz)。

B-1

¹ H NMR(400MHz,DMSO-d ₆ )δ5.97(s,1H),5.15(s,1H),4.85(s,2H),4.62(s,1H),4.35(s,1H),4.32–4.19(m,4H),4.08(s,1H),3.51(s,1H),3.38(s,1H),2.88(s,1H),2.74(s,9H),2.11(s,1H),2.05(s,1H),1.52(s,1H)。

¹³ C NMR(101MHz,DMSO-d ₆ )δ173.22(s),172.28(s),149.76(s),148.80(s),147.40(s),141.19(s),103.56(s),101.65(s),79.71(s),76.40(s),74.54(d,J＝3.1Hz),74.26(s),73.21(s),72.64(s),71.72(s),65.97(s),22.34(d,J＝7.3Hz)。

C-1

¹ H NMR(400MHz,DMSO-d ₆ )δ5.87(s,4H),5.63(s,4H),4.86(d,J＝29.0Hz,12H),4.57(s,4H),4.50(d,J＝15.1Hz,7H),4.45(d,J＝13.0Hz,2H),4.42(s,4H),4.43–4.03(m,24H),4.00(d,J＝9.6Hz,7H),3.78(s,2H),2.83(d,J＝5.9Hz,8H),2.74(s,35H),2.33(s,4H),1.81(s,4H),1.51(s,4H),1.40(s,4H),1.34(s,4H)。

¹³ C NMR(101MHz,DMSO-d ₆ )(s),141.19(s),103.56(s),103.05(s),101.65(s),79.71(s),76.40(s),74.54(d,J＝3.1Hz),74.26(s),73.15(d,J＝13.9Hz),72.64(s),71.72(s),65.97(s),22.34(d,J＝7.3Hz)。

D-1

¹ H NMR(400MHz,DMSO-d ₆ )δ5.70(s,4H),5.10(s,8H),4.28(s,2H),4.14(d,J＝14.3Hz,9H),4.04(s,3H),2.74(s,36H),1.86(s,4H),1.77(s,4H),1.61(s,4H)。(101MHz,DMSO-d ₆ )δ172.28(s),149.76(s),148.80(s),147.40(s),141.19(s),96.19(s),78.66(s),74.02(s),73.03–72.82(m),67.31(s),22.34(d,J＝7.3Hz)。

E-1

¹ H NMR(400MHz，DMSO-d ₆ )δ4.95(s，4H)，4.05(s，4H)，3.67(s，4H)，3.00(s，4H)，2.84(s，3H)，2.73(s，35H)，2.52(d，J＝11.1Hz，8H)，2.24(s，4H)，1.99(s，4H)。(101MHz，DMSO-d ₆ )δ177.60(s)，176.18(s)，151.54(s)，150.16(s)，149.00(s)，142.88(s)，74.65(s)，72.45(s)，68.38(s)，50.02(s)，34.51(s)，22.34(d，J＝7.3Hz)。

F-1

¹ H NMR(400MHz，DMSO-d ₆ )δ5.97(s，1H)，5.76(s，1H)，4.87(s，2H)，4.71(s，1H)，4.47-4.31(m，6H)，3.87(s，1H)，2.74(s，9H)，2.57(s，1H)，1.79(s，1H)，1.70(s，1H)，1.43(d，J＝8.0Hz，2H)。

(101MHz，DMSO-d ₆ )δ172.28(s)，149.76(s)，148.80(s)，147.40(s)，141.19(s)，103.56(s)，96.19(s)，79.71(s)，78.66(s)，74.27(s)，73.94(d，J＝18.7Hz)，73.50-72.99(m)，67.31(s)，22.34(d，J＝7.3Hz)。

G-1

¹ H NMR(400MHz，DMSO-d ₆ )δ6.71(s，1H)，5.33(s，1H)，4.78(d，J＝13.4Hz，3H)，4.24(s，1H)，4.20(s，1H)，4.05-3.65(m，5H)，3.96-3.65(m，3H)，3.94-3.61(m，3H)，2.89(s，1H)，2.74(s，10H)，2.14(s，1H)，2.05(s，1H)。

(101MHz，DMSO-d ₆ )δ176.18(s)，174.43(s)，172.28(s)，149.76(s)，148.80(s)，147.40(s)，141.19(s)，103.56(s)，78.66(s)，77.80(s)，74.27(s)，73.87(s)，73.10(s)，72.59(s)，71.44(d，J＝9.4Hz)，67.31(s)，22.34(d，J＝7.3Hz)。

example 2 Compounds A-D vs. CoCl ₂ Protective action for inducing hypoxia injury of nerve cell and cardiac muscle cell

The action and efficacy are described by taking a compound of a main characteristic system as an example:

wherein R = H or acetyl.

With no addition of CoCl ₂ Cell viability of induced neural cells PCl2 and cardiomyocytes H9C2 was negative control, and the observation of Compounds A-D on induced CoCl ₂ The specific implementation steps of the generated inhibition effect of the hypoxia injury of the nerve cells and the cardiac muscle cells are as follows: inoculating PC12 and H9C2 cells into MEM or DMEM complete culture solution, culturing in 96-well plate, incubating in constant temperature cell incubator for 24 hr, adding pre-dissolved CoCl-containing solution ₂ The serial compound solution is added into each hole after the injury solution is cultured for 2 hours, the concentration is 100 mu g/ml, and the incubator is used for incubation for 48 hours. After the incubation is finished, the cell survival rate is determined by adopting an MTT method, three times of experiments are carried out in parallel every time, and the experiments are repeated three times.

The experimental results are shown in tables 1 and 2, and show that the ligustrazine derivatives have good effects of protecting nerve cells and cardiac muscle cells and are used for CoCl ₂ The induction of the hypoxia injury of nerve cells and cardiac muscle cells has obvious inhibiting effect, and the protective effect of each group of derivatives is better than that of ligustrazine, wherein the derivatives have CoCl ₂ The best protective effect of group D1 in the nerve cells is induced; for CoCl ₂ The protective effect of group D1 was also best induced in cardiomyocytes.

TABLE 1 Tetramethylpyrazine derivatives vs CoCl ₂ Hypoxia-induced nerve cellsEffect of cellular injury

TABLE 2 Effect of ligustrazine derivatives on CoCl2 hypoxia-induced myocardial cell injury

Example 3 Effect of the Compounds A-D on platelet aggregation

New Zealand rabbits from the experimental animal center of Shandong province were selected as experimental animals, weighing 2-3kg, male. Fixing a rabbit on an operating table in a supine position, puncturing the heart with a needle to take blood, anticoagulating with sodium citrate, centrifuging at 1000rpm for 10min, and sucking the supernatant, namely platelet-rich plasma PRP; and centrifuging the rest blood again at 3000rpm for 20min to obtain supernatant which is the platelet poor plasma PPP. PRP platelet count adjusted to 4X 10 with PPP ⁸ And (4) the concentration is/ml. Adjusting to zero by PPP, adding PRP into a pen-turbid tube, adding the sample solution, bathing at 37 ℃ for 10min, respectively adding the series of compound solutions, measuring the platelet aggregation property on a platelet meter, recording the maximum platelet aggregation rate within 15min, and calculating the platelet aggregation inhibition rate.

Platelet aggregation inhibition (%) = [ 1-sample tube aggregation percentage/control tube aggregation percentage ] × 100%.

The results are shown in table 3, the ligustrazine derivatives can effectively inhibit platelet aggregation, and the performances of the ligustrazine derivatives are better than the effects of the ligustrazine, wherein the D0 ligustrazine derivative compound has the best effect.

TABLE 3 Effect of ligustrazine derivative Compounds on platelet aggregation in rabbits

The above examples are merely illustrative of the technical solutions of the present invention, and not limitative thereof; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.

Claims

1. A ligustrazine derivative is characterized in that: the ligustrazine derivative is as follows:

，

wherein R = H or acetyl.

2. A process for preparing a ligustrazine derivative according to claim 1, which comprises: the preparation method comprises the following steps:

3. The method for preparing a ligustrazine derivative according to claim 2, wherein: the step (2) is washed by saturated sodium bicarbonate and water, and dried by anhydrous sodium sulfate.

4. The use of the ligustrazine derivative of claim 1 for preparing a medicament for preventing or treating diseases of the nervous system and the cardiovascular system, wherein: the nervous system diseases are Alzheimer disease and Parkinson disease.

5. The use of the ligustrazine derivative according to claim 4 for the preparation of a medicament for the prevention or treatment of diseases of the nervous system and the cardiovascular system, characterized in that: the cardiovascular and cerebrovascular system diseases are cerebral ischemia diseases, cerebral anoxia, myocardial anoxia, thrombosis resistance and blood circulation improvement.

6. A pharmaceutical composition comprising the ligustrazine derivative of claim 1 and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition of claim 6, wherein: the pharmaceutical composition is in the form of tablet, oral liquid, pill, capsule, granule, syrup, powder, granule or tincture.

8. The pharmaceutical composition of claim 6, wherein: the pharmaceutical composition is a powder injection or an injection.