WO2018232805A1 - Puerarin derivative, preparation method thereof and use thereof for preventing or treating cardiovascular and cerebrovascular diseases or diabetes and complications thereof - Google Patents

Abstract

Provided are a puerarin derivative represented by formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, a preparation method therefor, and a use thereof for the prevention and treatment of cardiovascular and cerebrovascular diseases or diabetes and complications thereof. (III) may represent a single bond or a double bond; R1 represents an oxygen atom, a hydrogen atom, a hydroxyl group, an alkoxy group, or a halogen atom; and R2 and R3 individually represent a hydrogen atom, a hydroxyl group, a hydroxymethyl group, a halogen atom, -O(O)CR4, -OSOR4, -OSO2R4, or -O(O)PO2R4R5. A medical composition of the above compound and a salt thereof can be used for the prevention and treatment of cardiovascular and cerebrovascular diseases or diabetes and complications thereof. The compound can be used as a main ingredient for the preparation of a corresponding oral formulation and injection. The compound can also be used with other drugs in combined therapy regimes for treating cardiovascular and cerebrovascular diseases.

Description

Puerarin derivative, preparation method thereof and use thereof for preventing or treating cardiovascular and cerebrovascular diseases or diabetes and complications thereof Technical field

The present invention relates to the field of cardiovascular and vascular treatment of diabetes and the field of preparation of compounds, in particular, the present invention relates to the structure of novel derivatives of puerarin and a preparation method thereof, and the treatment and prevention of cardiovascular and cerebrovascular diseases or diabetes Use in complications.

Background technique

Puerarin is a flavonoid compound distributed in the leguminous plants of kudzu and kudzu root, and is one of the main active ingredients of the Chinese medicine Pueraria. Studies have shown that puerarin is a class of beta blockers and calcium antagonists, with pharmacological effects such as dilatation of coronary arteries, blood pressure lowering, antiarrhythmia, hypoglycemic, hypolipidemic, diastolic smooth muscle, inhibition of platelet aggregation and antioxidants. Clinically, it is mainly used for the treatment of cardiovascular and cerebrovascular diseases such as hypertension, coronary heart disease, angina pectoris, acute cerebral infarction and hyperviscosity. In addition, puerarin has a similar structure to the hypoglycemic substance phlorizin, both of which are flavonoids. Studies have shown that puerarin also has a certain blood sugar lowering function. Puerarin has important clinical application value because of its low toxicity, wide safety range and good curative effect.

At present, puerarin has shortcomings such as poor water solubility and low oral bioavailability during clinical use, which limits its clinical use. In particular, puerarin has a low water solubility of only 4.5 g/L. In clinical use, puerarin injection requires the addition of high concentrations of propylene glycol and lysine, arginine, histidine and nicotinamide. Solvents, and such cosolvents have limited effects, and may also cause toxic side effects such as allergic reactions and liver and kidney damage in the human body. Therefore, structural modification and modification of puerarin, development of new cardiovascular and cerebrovascular protective drugs and hypoglycemic drugs to enhance their water solubility, improve bioavailability and biological activity, have significant application value and market potential. The prior art also discloses various derivatives which are structurally modified or modified to enhance the water solubility of puerarin and corresponding preparation methods, such as CN101712676A, which discloses the design and synthesis of a highly water-soluble puerarin derivative prodrug such as puerarin phosphate. Salt, sulfonate, etc. to enhance the water solubility of the drug; CN103382203A discloses improving the water solubility and fat solubility of puerarin by enhancing the targeted modification of the drug; CN103694229A prepares a water-soluble and fat-soluble puerarin derivative. There is a glucopyranose in the 8th position of puerarin, and a phenolic hydroxyl group in the 7th and 4' positions. It is the active group of puerarin and is the concentrated site of puerarin structure modification.

Summary of the invention

In view of the problems existing in the prior art, in order to overcome the shortcomings and disadvantages of poor water solubility of puerarin, the present invention uses puerarin as a raw material, firstly by modifying its skeleton, reducing C ring α,β unsaturated double bonds, and destroying the molecular rigid structure. To improve its solubility; secondly, to further improve the molecular polarity and water solubility by further reducing the ketone carbonyl group to a hydroxyl group. Finally, several water-soluble puerarin derivatives were obtained. The invention provides a novel derivative of puerarin, which has good water solubility, It has high activity and low preparation cost, and can be used as a promising drug for preventing and treating cardiovascular and cerebrovascular diseases.

Specifically, the present invention relates to the following technical solutions:

First, it is an object of the present invention to provide a compound of the formula (I) or formula (II) or a pharmaceutically acceptable salt thereof,

可代表是单键，也可代表双键；R¹代表：氧原子、氢原子、羟基、烷氧基和卤素原子；R²和R³分别代表：氢原子、羟基、羟甲基、卤素原子，-O(O)CR⁴、-OSO₃R⁴、-O(O)PO₂R⁴R⁵；R⁴和R⁵可以独立选自氢原子、(C₁-C₄)烷基、氨基、金属离子，如钠离子、钾离子、镁离子、钙离子或锌离子等。among them,

It may represent a single bond or a double bond; R ¹ represents an oxygen atom, a hydrogen atom, a hydroxyl group, an alkoxy group and a halogen atom; and R ² and R ³ represent a hydrogen atom, a hydroxyl group, a hydroxymethyl group or a halogen atom, respectively. , -O(O)CR ⁴ , -OSO ₃ R ⁴ , -O(O)PO ₂ R ⁴ R ⁵ ; R ⁴ and R ⁵ may be independently selected from a hydrogen atom, a (C ₁ -C ₄ )alkyl group, an amino group Metal ions such as sodium ions, potassium ions, magnesium ions, calcium ions or zinc ions.

仅代表是单键。In a preferred embodiment of the invention,

Only the representative is a single button.

The puerarin derivative of the invention transforms the puerarin skeleton, reduces the C-ring α,β unsaturated double bond, destroys the molecular rigid structure, and improves the solubility thereof; further, by reducing the ketone carbonyl group to the hydroxyl group, Molecular polarity and water solubility. Illustratively, the puerarin derivative (compound 1-10) of the present invention has a significantly improved water solubility (4.5 g/L) compared to puerarin, wherein the compounds 5 to 10 have the best water solubility of 20-25 g/L. The solubility of the compounds 1 to 4 is slightly lower, both of which are about 10 g/L, but the solubility of puerarin is significantly improved. The puerarin derivative of the present invention can effectively improve the water solubility and improve the bioavailability while maintaining or improving the physiological activity of puerarin, and is suitable for injection.

In a preferred embodiment of the invention, the compound may be a compound selected from the following structures:

In a more preferred embodiment, the compounds of the invention are selected from the group consisting of compounds 5, 6, and 10.

In a preferred embodiment, the pharmaceutically acceptable salt of the compound of the present invention is also within the scope of the present invention, and the pharmaceutically acceptable salt of the compound is the compound and an inorganic base such as potassium hydroxide or sodium hydroxide. a salt formed by potassium carbonate, sodium carbonate, calcium chloride, calcium acetate or magnesium chloride, or a salt formed with an organic base such as tromethamine, aminoethanol, lysine or arginine.

Secondly, a second object of the present invention is to provide a process for producing the above compound or a pharmaceutically acceptable salt thereof, which comprises the following reaction,

可代表是单键，也可代表双键；R¹代表：氧原子、氢原子、羟基、烷氧基和卤素原子。Catalytic hydrogenation of a compound represented by formula (III) or (V) to give a compound of formula (IV) or formula (VI), wherein

It may represent a single bond or a double bond; R ¹ represents an oxygen atom, a hydrogen atom, a hydroxyl group, an alkoxy group, and a halogen atom.

可代表是单键，也可代表双键；R¹代表：氧原子、氢原子和羟基。In a preferred embodiment, the compound represented by formula (IV) or formula (VI) is prepared by using puerarin (III) or formula 7, 2"-anhydropuerarin (V) as a starting material, respectively. Preparation: puerarin or dehydrated puerarin under palladium carbon catalysis in methanol or ethyl acetate solution, by controlling the reaction time to obtain different degrees of reduction of the compound of formula (IV) or formula (VI), wherein

It may represent a single bond or a double bond; R ¹ represents an oxygen atom, a hydrogen atom and a hydroxyl group.

It is still another object of the present invention to provide a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt thereof. It can be used either as a monomer or as a mixture of isomers. The excipients used in the pharmaceutical compositions may be in solid or liquid form. Solid form preparations include powders, tablets, dispersion granules, capsules, pills, and suppositories. Powders and tablets may contain from about 5% to about 95% of the active ingredient. Suitable solid adjuvants can be magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, pills and capsules are solid dosage forms suitable for oral administration. The liquid form preparations include solutions, suspensions and emulsions, and examples thereof are aqueous solutions for parenteral injection or water-propylene glycol solutions, or oral solutions in which sweeteners and contrast agents are added. In addition, it can also be used as a small water needle for injection, a freeze-dried powder for injection, a large infusion or a small infusion.

Another object of the present invention is to provide a compound of the above formula (I) or formula (II) or a pharmaceutically acceptable salt thereof for use in the preparation of a medicament for preventing and/or treating cardiovascular and cerebrovascular diseases or a hypoglycemic agent. These cardiovascular and cerebrovascular diseases include high blood Pressure, angina, heart failure, myocardial infarction, stroke and cerebral hemorrhage, in addition to diabetes and its complications.

Still another object of the present invention is to include a compound of the above formula (I) or formula (II) or a pharmaceutically acceptable salt thereof in combination with at least one other cardiovascular disease drug or hypoglycemic agent. The atomic composition or structure of other cardiovascular disease drugs or hypoglycemic drugs covered is different from the compound of formula (I) or formula (II). Other cardiovascular disease agents that can be used in combination with the novel compounds of the invention include those having antithrombotic, antiplatelet aggregation, antiatherosclerosis, anti-restenosis and/or anticoagulant, vasodilating activity, and diabetes and Its complications.

The present invention achieves the following beneficial effects:

(1) The present invention uses puerarin as a raw material to modify the skeleton thereof to reduce the C-ring α,β unsaturated double bond, thereby destroying the rigid structure of the molecule and improving its solubility; secondly, by further reducing the ketone carbonyl group to a hydroxyl group, Molecular polarity and water solubility, and finally obtained a number of well-water soluble puerarin derivatives.

(2) It was confirmed by animal experiments that the composition of the present invention can increase the coronary blood flow of guinea pigs, relax the nociceptin-induced arterial contraction, and damage the rats with focal cerebral ischemia reperfusion injury. It has a protective effect and effectively maintains or enhances the physiological activity of puerarin while improving the bioavailability of puerarin.

(3) The compound of the invention is simple and easy to prepare, has high compound activity and low preparation cost, and can be used as a promising drug for preventing and treating cardiovascular and cerebrovascular diseases.

DRAWINGS

Figure 1: Effect of norepinephrine on arterial contraction - blank control

Figure 2: Relaxation of puerarin on arterial contraction caused by norepinephrine

Figure 3: Relaxation of compound 10 on arterial contraction caused by norepinephrine

Figure 4: Relaxation of arterial contractions induced by norepinephrine in a mixture of compounds 5 and 6

Detailed ways

The present invention is further described in detail below, but the embodiments of the present invention are not limited thereto. It will be apparent to those skilled in the art that various modifications and changes can be made in the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and scope of the present invention are intended to be included in the scope of the present invention. within.

Example 1: The structure of the compound of the present invention is as follows:

The pharmaceutically acceptable salt of the above compound is a salt of the compound with an inorganic base such as potassium hydroxide, sodium hydroxide, potassium carbonate, sodium carbonate, calcium chloride, calcium acetate or magnesium chloride, or with an organic base such as an amino group. A salt formed by butyl triol, aminoethanol, lysine or arginine.

Example 2: Preparation of Compounds 1 and 2

The compound puerarin (III) was weighed and placed in a dry round bottom flask, and dissolved in ethyl acetate. Then, 10% palladium carbon was added thereto, and the reaction was stirred at room temperature for 6 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, palladium carbon was filtered off. The solvent was distilled off, and the obtained product was subjected to a flash reverse phase column, and the mobile phase was eluted with a gradient of methanol/water, and finally, the solvent was evaporated under reduced pressure to give Compounds 1 and 2 as white solid.

Compound 1: MS m/z: 419 (M + 1); ¹ H NMR (400 MHz, MeOH) δ 7.75 (d, J = 8.8 Hz, 1H), 7.08 (d, J = 8.6 Hz, 2H), 6.75 (d, J = 8.5 Hz, 2H), 6.54 (d, J = 8.8 Hz, 1H), 4.93 - 4.88 (m, 2H), 4.63 - 4.52 (m, 2H), 4.04 (s, 1H), 3.87 ( Dd, J = 11.9, 1.9 Hz, 1H), 3.73 (dd, J = 11.9, 4.9 Hz, 1H), 3.50 - 3.36 (m, 3H).

Compound 2: MS m/z: 419 (M + 1); ¹ H NMR (400 MHz, Me OD) δ 7.75 (d, J = 8.8 Hz, 1H), 7.12 (d, J = 8.5 Hz, 2H), 6.74 (d, J = 8.5 Hz, 2H), 6.55 (d, J = 8.8 Hz, 1H), 4.95 - 4.89 (m, 2H), 4.64 - 4.54 (m, 2H), 4.01 (s, 1H), 3.87 ( Dd, J = 12.0, 2.0 Hz, 1H), 3.77 - 3.70 (m, 1H), 3.50 - 3.37 (m, 3H).

Example 3: Preparation of Compounds 3 and 4

Weighing compound dehydrated puerarin (V) was placed in a dry round bottom flask, dissolved in ethyl acetate, followed by addition of 10% palladium on carbon, and the reaction was stirred at room temperature for 6 hours. The progress of the reaction was monitored by TLC. Carbon, the solvent was distilled off, the obtained product was applied to a flash reversed phase column, and the mobile phase was eluted with a gradient of methanol/water, and finally, the solvent was evaporated under reduced pressure to give white solid compounds 3 and 4.

Compound 3: MS m/z: 401 (M + 1); ¹ H NMR (600 MHz, CD ₃ OD) δ 7.92 (d, J = 8.6 Hz, 1H), 7.08 (d, J = 8.6 Hz, 2H) , 6.77 (d, J = 8.6 Hz, 2H), 6.68 (d, J = 8.6 Hz, 1H), 5.18 (d, J = 3.3 Hz, 1H), 4.70 - 4.55 (m, 3H), 4.00 - 3.97 ( m, 2H), 3.84 (dd, J = 12.1, 2.5 Hz, 1H), 3.64 (dd, J = 12.1, 5.7 Hz, 1H), 3.60 (t, J = 9.4 Hz, 1H), 3.35 (ddd, J =9.3, 5.7, 2.3 Hz, 1H).

Compound 4: MS m/z: 401 (M + 1); ¹ H NMR (600 MHz, CD ₃ OD) δ 7.92 (d, J = 8.6 Hz, 1H), 7.09 (d, J = 8.6 Hz, 2H) , 6.74 (d, J = 8.6 Hz, 2H), 6.68 (d, J = 8.5 Hz, 1H), 5.18 (d, J = 3.3 Hz, 1H), 4.71 - 4.63 (m, 3H), 3.98 (dd, J=9.5, 4.8 Hz, 1H), 3.88 (dd, J=7.4, 5.3 Hz, 1H), 3.84 (dd, J=12.0, 2.4 Hz, 1H), 3.64 (dd, J=12.1, 5.8 Hz, 1H) ), 3.59 (t, J = 9.4 Hz, 1H), 3.35 (ddd, J = 8.0, 5.8, 2.4 Hz, 1H).

Example 4: Preparation of Compound 5-8

The compound puerarin (III) was weighed and placed in a dry round bottom flask, dissolved in ethyl acetate, and then 10% palladium on carbon was added thereto. The reaction was stirred at room temperature for 48 hours, and the progress of the reaction was monitored by TLC. After the reaction was completed, palladium carbon was filtered off. The solvent was distilled off, and the obtained product was subjected to a silica gel reversed phase column, and the mobile phase was eluted with a gradient of methanol/dichloromethane, and the solvent was evaporated under reduced pressure to give a mixture of compounds 5, 6, 7 and 8. Finally, through HPLC reverse phase column, the mobile phase was acetonitrile/water, and the pure products of compounds 5, 6, 7 and 8 were obtained, respectively.

Compound 5: MS m/z: 421 (M + 1); ¹ H NMR (400 MHz, Me OD) δ 7.24 (d, J = 8.6 Hz, 1H), 7.07 (d, J = 8.5 Hz, 2H), 6.73 (d, J = 8.5 Hz, 2H), 6.46 (d, J = 8.5 Hz, 1H), 4.77 (d, J = 7.2 Hz, 1H), 4.29 (dd, J = 11.1, 3.6 Hz, 1H), 4.17 (dd, J = 11.0, 8.6 Hz, 1H), 4.01 - 3.91 (m, 1H), 3.84 (dd, J = 11.8, 1.8 Hz, 1H), 3.74 (dd, J = 12.1, 5.0 Hz, 1H), 3.46 (t, J = 7.3 Hz, 2H), 3.40-3.36 (m, 1H), 3.01 (td, J = 8.1, 3.5 Hz, 1H).

Compound 6: MS m/z: 421 (M + 1); ¹ H NMR (400 MHz,MeOD) δ 7.12 (d, J = 8.6 Hz, 2H), 7.08 (d, J = 8.5 Hz, 1H), 6.76 (d, J = 8.6 Hz, 2H), 6.44 (d, J = 8.4 Hz, 1H), 4.60 (d, J = 2.2 Hz, 1H), 4.47 (dd, J = 11.9, 10.7 Hz, 1H), 4.22 (dd, J = 10.3, 2.6, 1H), 3.99 (t, J = 8.9 Hz, 1H), 3.85 (dd, J = 12.1, 2.2 Hz, 1H), 3.75 (dd, J = 12.1, 4.9 Hz, 1H) ), 3.49 (t, J = 7.3 Hz, 2H), 3.40-3.36 (m, 1H), 3.11 (dt, J = 12.00, 3.2 Hz, 1H).

Compound 7: MS m/z: 421 (M + 1); ¹ H NMR (400 MHz,MeOD) δ 7.12 (d, J = 8.5 Hz, 2H), 7.09 (d, J = 8.4 Hz, 1H), 6.76 (d, J = 8.5 Hz, 2H), 6.44 (d, J = 8.4 Hz, 1H), 4.61 (d, J = 2.1 Hz, 1H), 4.45 (dd, J = 12.0, 10.5 Hz, 1H), 4.25 (dd, J = 10.3, 2.4 Hz, 1H), 3.99 (t, J = 8.7 Hz, 1H), 3.85 (dd, J = 12.0, 2.1 Hz, 1H), 3.76 (dd, J = 12.0, 4.8 Hz, 1H), 3.48 (t, J = 8.3 Hz, 2H), 3.43 - 3.37 (m, 1H), 3.15 (dt, J = 12.1, 3.2 Hz, 1H).

Compound 8: MS m/z: 421 (M + 1); ¹ H NMR (400 MHz, MeOH) δ 7.21. (d, J = 8.6 Hz, 1H), 7.11 (d, J = 8.3, 2H), 6.73 ( d, J = 8.3 Hz, 2H), 6.45 (dd, J = 8.4, 3.1 Hz, 1H), 4.71 (d, J = 6.8 Hz, 1H), 4.30 (dd, J = 11.1, 3.7 Hz, 1H), 4.22 (dd, J = 11.1, 7.8 Hz, 1H), 3.96 (t, J = 8.9 Hz, 1H), 3.85 (dd, J = 12.0, 2.2 Hz, 1H), 3.76 (dd, J = 12.1, 4.7 Hz) , 1H), 3.48 (t, J = 8.1 Hz, 2H), 3.43 - 3.37 (m, 1H), 2.99 (td, J = 7.4, 3.7 Hz, 1H).

Example 5: Preparation of Compounds 9 and 10

The compound 3 was weighed and placed in a dry round bottom flask, dissolved in ethyl acetate, and then added with 10% palladium carbon. The reaction was stirred at room temperature for 36 hours. The reaction was monitored by TLC. After completion of the reaction, the palladium carbon was filtered off and the solvent was evaporated. The obtained product was passed through a silica gel reverse phase column, and the mobile phase was eluted with a gradient of methanol/dichloromethane, and the solvent was evaporated under reduced pressure to give a mixture of compounds 9 and 10. Finally, through HPLC reverse phase column, the mobile phase was acetonitrile/water, and the pure products 9 and 10 were obtained, respectively.

Compound 9: MS m/z: 403 (M + 1); ¹ H NMR (600 MHz, CD ₃ OD) δ 7.36 (d, J = 8.3 Hz, 1H), 7.08 (d, J = 8.5 Hz, 2H) , 6.73 (d, J = 8.6 Hz, 2H), 6.53 (d, J = 8.3 Hz, 1H), 5.07 (d, J = 3.3 Hz, 1H), 4.80 (d, J = 7.3 Hz, 1H), 4.52 (dd, J = 4.7, 3.4 Hz, 1H), 4.35 (dd, J = 11.1, 3.7 Hz, 1H), 4.27 (dd, J = 11.1, 8.4 Hz, 1H), 3.93 (dd, J = 9.5, 4.8 Hz, 1H), 3.84 (dd, J = 12.0, 2.3 Hz, 1H), 3.60 (dd, J = 12.1, 6.1 Hz, 1H), 3.55 (t, J = 9.4 Hz, 1H), 3.34 - 3.32 (m) , 1H), 3.03 (td, J = 7.9, 3.6 Hz, 1H).

Compound 10: MS m/z: 403 (M + 1); ¹ H NMR (400 MHz, CD ₃ OD) δ 7.09 (d, J = 8.3 Hz, 2H), 7.08 (d, J = 8.4 Hz, 1H) , 6.68 (d, J = 8.5 Hz, 2H), 6.43 (d, J = 8.1 Hz, 1H), 4.92 (d, J = 3.1 Hz, 1H), 4.52 - 4.40 (m, 3H), 4.35 (dd, J=4.8, 3.1 Hz, 1H), 4.21–4.10 (m, 1H), 3.76 (dd, J=9.4, 4.7 Hz, 1H), 3.68–3.55 (m, 1H), 3.42-3.38 (m, 1H) , 3.13 (ddd, J = 8.9, 5.3, 1.9 Hz, 1H), 3.05 (dt, J = 12.3, 2.9 Hz, 1H).

Compound 1-10 was tested at room temperature (25 ° C) for water solubility. The water solubility of puerarin derivatives (compounds 1-10) was significantly higher than that of puerarin (4.5 g/L), and the solubility of compounds 5 to 10 was the best. It is 20-25 g/L, and the water solubility of the compounds 1 to 4 is slightly lower, both of which are about 10 g/L, and the compound 10 and the compound 5 and 6 having good water solubility are subjected to subsequent tests.

Example 6: Effect of compound of formula (I) or formula (II) on coronary flow in isolated guinea pigs

1. Material:

Animal: Guinea pig

Instruments: thermostat, oxygen supply system, constant pressure device, aortic cannula, surgical instruments

Drugs: Puerarin injection, Compound 10, Compounds 5 and 6 were dissolved in sterile saline to 10 mg/ml and 5 mg/ml, respectively, and the intraperitoneal injection and gavage volume was 0.5 mL/100 g.

2. Method:

(1) guinea pigs, half male and half female, weighing 250-350g, were randomly divided into normal group, puerarin group (50mg/kg), compound 10 low dose group (25mg/kg,), compound 10 high dose group (50mg/kg). , low dose group of compound 5 and 6 mixture (25mg/kg), compound 5 and 6 mixture high dose group (50mg/kg) 4-5 per group, respectively, intraperitoneal injection, once a day for 10 days.

In addition, the normal group, the compound 10 high dose group (50 mg/kg), and the compound 5 and 6 mixture high dose group (50 mg/kg) were intragastrically administered for 7 days.

(2) Using the Langendorff method to observe the effect of the compound on coronary flow, adjust the constant temperature and constant pressure perfusion apparatus: temperature 37 ± 0.5 ° C, oxygen 2-3 / s, pressure 40-60 cm water column.

(3) After 30 minutes of the last administration, 20% urethane (0.5ml/100g) was anesthetized, cut the chest skin, cut the sternum from the xiphoid, quickly open the chest, expose the heart, gently hold the heart with the left hand, cut up and down The vena cava, the pulmonary artery, the aorta, and the tissues surrounding the heart are excised from the heart. Place in an oxygenated 4 ° C petri dish in Rockwell, gently squeeze the heart with your fingers, drain the residual blood in the heart, remove the pericardium, and find the aorta. The lobes, trachea, etc., which are connected to the heart, are trimmed, and the aorta is hung on the aortic cannula to provide ex vivo conditions. After 10 minutes of stabilization, the coronary flow per minute was measured continuously for 20 min.

3. Results:

Puerarin group (50mg/kg), compound 10 low dose group (25mg/kg,), compound 10 high dose group (50mg/kg), compound 5 and 6 mixture low dose group (25mg/kg), compounds 5 and 6 The high-dose group (50 mg/kg) increased coronary blood flow in guinea pigs compared with the normal group. The high dose group (50 mg/kg) of the compound 5 and 6 mixture was statistically significant compared with the normal group (intraperitoneal injection *P0.05; gavage **P0.01). Compared with the puerarin group, the high dose group of compound 5 and 6 (50 mg/kg) increased the coronary blood flow of guinea pigs more obviously. The specific results are shown in Table 1.

Table 1: Effect of puerarin derivatives on coronary flow in isolated guinea pigs

Compared with the normal group of the same administration route: ^* P0.05, ^** P0.01.

4. Conclusion: The mixture of puerarin derivatives 5 and 6 increased the coronary blood flow of guinea pigs more obviously.

Example 7: Effect of a compound of formula (I) or formula (II) on aortic rings in rabbits

1. Method:

Take the rabbit, stun the scalp, let the carotid artery bleed to death, quickly open the chest to remove the thoracic aorta, place the cold kein's solution, separate the surrounding tissue, cut into a 0.5mm ring, reserve, and adjust the artery ring In the bath (with 10ml of Heinz's solution inside), and linked with the pressure transducer (front load is set to 2g), the electrical signal is transmitted to the computer. 37 ° constant temperature, pass O ₂ , after 2 hours of stabilization, give 10 ^-6 mmol / L norepinephrine, after the peak of arterial contraction, add the cumulative concentration of puerarin, compound 10, compound 5 and 6 respectively 0.1 mg/ml, 0.5 mg/ml, 1 mg/ml, and 2 mg/ml (the same volume of physiological saline was added to the saline group), and the relaxation of the blood vessels was observed.

2. Results

The specific results are shown in Figures 1 to 4, in which the ordinate represents the arterial vasoconstriction tension, and the higher the value, the greater the vasoconstriction force. The abscissa represents time. From the results, it was found that the compound 10 and the mixture of the compounds 5 and 6 have a relaxing effect on norepinephrine-induced arterial contraction.

Example 8: Pharmacodynamic observation of the protective effect of the compound of formula (I) or formula (II) on focal cerebral ischemia reperfusion injury in rats

1. Method:

90 SD male rats weighing 280-320 g were randomly divided into 5 groups. The rats were fasted for 12 hours before the experiment. 2% pentobarbital sodium solution 0.2ml/100g was intraperitoneally injected into the anesthesia, fixed in the supine position, and the model of focal cerebral ischemia-reperfusion injury was made by suture method (ischemia for 2 hours and reperfusion for 24 hours), immediately after ischemia. Intravenous administration (puerarin, compound 10, compound 5 and 6 were 100 mg/kg, sham group and model group were given the same volume of NS; 402 was intraperitoneally injected), and neurological score was observed after 24 hours; cardiac blood sampling (4000 rpm / min centrifuge for 10 min, take serum for use), decapitate the brain, freeze the refrigerator for 30 min, cut the rat brain into 6 pieces, set the TTC dye solution, 37 ° water bath for 30 min, take out the photo, use Jetta 801 form The analysis software calculates the percentage of infarcts. The SOD and MDA contents were measured by biochemical methods.

2. Results

The specific experimental results are shown in Table 2 and Table 3:

Table 2: Protective effects of compound 10, compound 5 and mixture 6 on cerebral ischemia-reperfusion injury

Note: ^### p<0.001, vs sham operation group; ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001, vs model group.

Table 3: Effects of Compound 10, Compounds 5 and 6 on serum SOD and MDA in cerebral ischemia-reperfusion injury

Note: ^### p<0.001, vs sham operation group; ^*** p < 0.001, vs model group.

3. Conclusion

(1) Compound 10, compound 5 and mixture 6 can reduce the area of cerebral infarction caused by focal cerebral ischemia-reperfusion injury in rats and reduce the neurological score.

(2) Compound 10, compound 5 and mixture 6 can increase serum SOD content and reduce MDA content caused by focal cerebral ischemia-reperfusion injury.

Claims (10)

a compound of the formula (I) or formula (II) or a pharmaceutically acceptable salt thereof;

among them,

It may represent a single bond or a double bond; R ¹ represents one of an oxygen atom, a hydrogen atom, a hydroxyl group, an alkoxy group and a halogen atom; R ² and R ³ represent a hydrogen atom, a hydroxyl group, and a hydroxyl group, respectively. a group, a halogen atom, -O(O)CR ⁴ , -OSO ₃ R ⁴ , -O(O)PO ₂ R ⁴ R ⁵ , R ⁴ and R ⁵ may be independently selected from a hydrogen atom, (C ₁ -C ₄ ) Alkyl, sodium, potassium, magnesium, calcium, zinc or amino. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is a compound selected from the following structures:

The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2, wherein the pharmaceutically acceptable salt of the compound is a salt of the compound with an inorganic base or with an organic base. a salt formed, preferably, the inorganic base is selected from the group consisting of potassium hydroxide, sodium hydroxide, potassium carbonate, sodium carbonate, calcium chloride, calcium acetate or magnesium chloride, and the organic base is selected from the group consisting of tromethamine, aminoethanol, lysine or Arginine. A process for the preparation of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, which comprises the following reaction,

Catalytic hydrogenation of a compound represented by formula (III) or (V) to give a compound of formula (IV) or formula (VI), wherein

It may represent a single bond or a double bond; R ¹ represents an oxygen atom, a hydrogen atom, a hydroxyl group, an alkoxy group, and a halogen atom. The preparation method according to claim 4, wherein the compound represented by the formula (IV) or the formula (VI) is prepared by using puerarin (III) or 7, 2"-dehydrated puerarin (V), respectively. The starting material is prepared by the following method: puerarin or 7,2"-dehydrated puerarin under the palladium carbon catalysis in methanol or ethyl acetate solution, by controlling the reaction time to obtain different degrees of reduction as in formula (IV) Or a compound represented by the formula (VI). A pharmaceutical composition comprising the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, used as a monomer or as a mixture of isomers. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers, excipients, solubilizers and/or diluents. The pharmaceutical composition according to claim 6 or 7, wherein the pharmaceutical composition is a solid oral preparation, a liquid oral preparation or an injection. The pharmaceutical composition according to any one of claims 6 to 8, wherein the pharmaceutical composition is a tablet, a dispersible tablet, an enteric coated tablet, a chewable tablet, an orally disintegrating tablet, a capsule, a sugar coating agent, and a granule , dry powder, oral solution, small water needle for injection, freeze-dried powder for injection, large infusion or small infusion. Use of a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 6 to 9 for the preparation of prophylaxis and/or treatment Application in cardiovascular and cerebrovascular diseases and/or diabetes and its complications, Preferably, the cardiovascular and cerebrovascular diseases are selected from the group consisting of atherosclerosis, restenosis, hypertension, angina pectoris, heart failure, myocardial infarction, stroke and cerebral hemorrhage; Preferably, the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 6 to 9 and other cardiovascular and cerebrovascular diseases therapeutic drugs or other diabetes The combined use of therapeutic drugs in the clinic.

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