Polyphenolic compound as well as preparation method and application thereof

Abstract

The invention discloses a polyphenol compound as well as a preparation method and application thereof. The invention specifically provides a polyphenol compound as shown in a formula X, and a stereoisomer or pharmaceutically acceptable salt thereof. The compound provided by the invention has a remarkable protection effect on an OGD / R-induced in-vitro cerebral ischemia model, can remarkably inhibit inflammation and apoptosis reaction of neurons, can reduce the cerebral infarction volume of a cerebral ischemia mouse, is reduced in polarity, and improves the oral bioavailability and the proportion of passing through a blood-brain barrier; # imgabs0 #

Description

[0001] This application claims priority from Chinese patent application No. 202311345891X filed on October 16, 2023. This application cites the entire text of the above Chinese patent application. Technical Field

[0002] The present invention belongs to the field of biomedicine, and specifically relates to a polyphenol compound, a preparation method and application thereof, and in particular to application thereof in the preparation of a medicine for preventing and treating cerebral ischemia-related diseases. Background Art

[0003] Cerebral ischemia refers to a series of symptoms caused by insufficient blood supply to the brain, which is difficult to meet the metabolic needs of brain tissue. Clinically, it can manifest as dizziness, headache, limb numbness or temporary loss of consciousness. In severe cases, it can cause irreversible damage to brain function or even death. Diseases related to cerebral ischemia include transient ischemic attack (TIA), ischemic stroke (cerebral infarction), moyamoya disease, chronic cerebral ischemia, etc. Cerebral ischemia is one of the three major diseases that threaten human health, with high morbidity, high recurrence rate, high disability rate and high mortality rate. Ischemic stroke accounts for 60% to 80% of stroke, but there is still a lack of effective drugs for the treatment of ischemic stroke in clinical practice, so research on cerebral ischemia is crucial. After cerebral ischemia, the ischemic area will edema and secrete a large number of inflammatory factors; because neurons are the most sensitive, a large number of neurons in the infarction center and its surrounding areas die. Therefore, how to protect neurons and reduce neuronal death is also a major focus of research on neural repair after cerebral ischemia.

[0004] Although there are brain protective agents in the prior art, such as edaravone, the drug is not effective and has many serious adverse reactions, such as acute renal failure, abnormal liver function, thrombocytopenia, etc. Therefore, other drugs that can effectively treat cerebral ischemia are urgently needed. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the defect of the lack of effective drugs for the prevention and treatment of ischemic stroke in the prior art, and to provide a polyphenol compound, a preparation method and application thereof. The compound of the present invention has a good protective effect on the in vitro cerebral ischemia model induced by OGD / R, can inhibit the inflammation and apoptosis of neurons, can reduce the volume of cerebral infarction in cerebral ischemic mice, and has reduced polarity, thereby improving oral bioavailability and the proportion of crossing the blood-brain barrier.

[0006] In order to solve the above technical problems, the first aspect of the present invention provides a polyphenol compound as shown in Formula X, its stereoisomers or pharmaceutically acceptable salts thereof:

[0007]

[0008] Where R1 is C1-6 Alkyl, C 6-14 Aryl or 5- to 6-membered heteroaryl; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R2 and R3 are hydroxyl protecting groups or hydrogen;

[0009] R2 and R3 are independently a hydroxyl protecting group or a hydrogen atom; the hydroxyl protecting group is C 1-6 Acyl.

[0010] In some embodiments of the present invention, the polyphenol compound represented by X is a compound represented by formula I:

[0011]

[0012] Where R1 is C 1-6 Alkyl or 5- to 6-membered heteroaryl; the heteroatoms of the 5- to 6-membered heteroaryl are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0013] In some embodiments of the present invention, the C 1-6 The alkyl group is ethyl; or the number of heteroatoms of the 5- to 6-membered heteroaryl group is 1 or 2, for example, pyridin-3-yl or pyridin-4-yl.

[0014] In some embodiments of the present invention, the C 1-6 The alkyl group is methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl or isobutyl, preferably methyl, ethyl, isopropyl, n-propyl or isobutyl.

[0015] In some embodiments of the present invention, the number of heteroatoms of the 5- to 6-membered heteroaryl group is 1 or 2, for example, pyridyl.

[0016] In some embodiments of the present invention, the C 6-14 Aryl is phenyl or naphthyl, for example phenyl.

[0017] In some embodiments of the present invention, the C 1-6 The acyl group is formyl, acetyl, isopropionyl, n-propionyl, n-butyryl, tert-butyryl or isobutyryl, preferably acetyl.

[0018] Preferably, the compound of formula X is a compound represented by formula X-1:

[0019]

[0020] R1, R2 and R3 are independently as described in any embodiment of the present invention.

[0021] Preferably, the compound of formula I has the following structure:

[0022]

[0023] R1 is defined as described in any embodiment of the present invention.

[0024] Preferably, R1 is C 1-6 Alkyl, for example methyl, ethyl, isopropyl, n-propyl or isobutyl.

[0025] In some preferred embodiments of the present invention, the compound as shown in Formula I is any of the following compounds:

[0026]

[0027] In order to solve the above technical problems, the second aspect of the present invention provides a polyphenol compound, the general formula of the compound is shown in Formula XI:

[0028]

[0029] Where R1' is C 1-6 Alkyl, C 2-6 Alkenyl, C 6-14 Aryl, 5- to 6-membered heteroaryl; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0030] R' is a hydroxyl protecting group, and R2' and R3' are hydroxyl protecting groups or hydrogen atoms.

[0031] In some preferred embodiments of the present invention, the general formula of the compound represented by formula XI is as shown in formula II:

[0032]

[0033] Where R1' is C 2-6 Alkenyl or 5- to 6-membered heteroaryl; the heteroatoms of the 5- to 6-membered heteroaryl are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0034] R' is a hydroxyl protecting group.

[0035] In some preferred embodiments of the present invention, the R' is benzyl; the C 2-6 The alkenyl group is vinyl; the number of heteroatoms of the 5- to 6-membered heteroaryl group is 1 or 2, for example, pyridin-3-yl or pyridin-4-yl.

[0036] In some preferred embodiments of the present invention, the R' is a benzyl group; the R2' and R3' are acetyl groups or hydrogen atoms; the C 2-6The alkenyl group is vinyl, prop-1-en-2-yl, (E)-prop-1-en-1-yl or 2-methylprop-1-en-1-yl; the aryl group is phenyl; the number of heteroatoms of the 5- to 6-membered heteroaryl group is 1 or 2, for example, pyridin-3-yl or pyridin-4-yl.

[0037] In some embodiments of the invention, the halogen is chlorine or bromine, such as bromine.

[0038] In some embodiments of the present invention, in R2' and R3', the hydroxyl protecting group is C 1-6 The acyl group is preferably formyl, acetyl, isopropionyl, n-propionyl, n-butyryl, tert-butyryl or isobutyryl, for example acetyl.

[0039] In some embodiments of the present invention, the C 2-6 Alkenyl is vinyl, prop-1-en-2-yl, (E)-prop-1-en-1-yl or 2-methylprop-1-en-1-yl, for example vinyl.

[0040] In some embodiments of the present invention, the number of heteroatoms of the 5- to 6-membered heteroaryl group is 1 or 2, for example, pyridyl.

[0041] In some embodiments of the present invention, the C 6-14 Aryl is phenyl or naphthyl, for example phenyl.

[0042] In some embodiments of the present invention, the C 1-6 The alkyl group is methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl or isobutyl, preferably methyl, ethyl, isopropyl, n-propyl or isobutyl, for example methyl.

[0043] Preferably, the structure of the compound of formula XI is as follows:

[0044]

[0045] Preferably, the structure of the compound of formula II is as follows:

[0046]

[0047] R1' and R' are independently as described in any embodiment of the present invention.

[0048] In some more preferred embodiments of the present invention, the compound as shown in Formula II is any of the following compounds:

[0049]

[0050]

[0051] In order to solve the above technical problems, the third aspect of the present invention provides a polyphenol compound as shown in formula III:

[0052]

[0053] Wherein, R1' and R' are defined as above; R" is a hydroxyl protecting group, such as acetyl.

[0054] Preferably, the structure of the compound of formula III is as follows:

[0055]

[0056] The compound III is preferably the following structure:

[0057]

[0058] In order to solve the above technical problems, the fourth aspect of the present invention provides a method for preparing the compound represented by formula X, which is any of the following schemes:

[0059] Solution 1:

[0060] The method comprises the following steps: under the catalysis of catalyst 1 and in the presence of hydrogen, a compound as shown in formula XI is subjected to a reaction of removing the protecting group R' in an organic solvent to obtain a compound as shown in formula X;

[0061]

[0062] Where R1' is C 1-6 Alkyl, C 6-14 Aryl or 5- to 6-membered heteroaryl;

[0063] R', R2', R3', the C 1-6 Alkyl, C 6-14 Aryl, 5- to 6-membered heteroaryl, R1, R2 and R3 are independently as described in any embodiment of the present invention;

[0064] The R' is preferably a benzyl group; the R2' and R3' are preferably an acetyl group or hydrogen;

[0065] Scheme 2: The method comprises the following steps: Under the catalysis of catalyst 1 and in the presence of hydrogen, the compound represented by formula XI is subjected to a reaction of removing the protecting group R' and reducing the olefin group in an organic solvent to obtain a compound represented by formula X.

[0066]

[0067] Where R1' is C 2-6 alkenyl;

[0068] R', R2', R3', the C 2-6 Alkenyl, R1, R2 and R3 are independently as described in any embodiment of the present invention;

[0069] The R' is preferably benzyl; wherein the R2' and R3' are preferably acetyl or hydrogen;

[0070] In some more preferred embodiments of the present invention, the catalyst 1 is 20% palladium hydroxide on carbon.

[0071] In some more preferred embodiments of the present invention, the organic solvent is methanol.

[0072] The present invention also provides a method for preparing the compound shown in formula I as described in the first aspect of the present invention, the method comprising the following steps: under the catalysis of catalyst 1, the compound shown in formula II is subjected to a reaction of removing the hydroxyl protecting group R' in an organic solvent to obtain the compound shown in formula I.

[0073] Preferably, the method comprises the following steps: under the catalysis of catalyst 1 and in the presence of hydrogen, the compound shown in formula II is subjected to a reaction of removing the protecting group R' in an organic solvent to obtain a compound shown in formula I, wherein R1' is a 5- to 6-membered heteroaryl group; for example, pyridin-3-yl or pyridin-4-yl.

[0074] Alternatively, the method comprises the following steps: in the presence of hydrogen and under the catalysis of catalyst 1, the compound shown in formula II is subjected to a reaction of removing the protecting group R' and reducing the olefin group in an organic solvent to obtain a compound shown in formula I, wherein R1' is C 2-6 Alkenyl, such as vinyl.

[0075] Wherein, the R' is preferably benzyl.

[0076] Preferably, R2' and R3' are acetyl groups or hydrogen atoms; wherein R1' is methyl, C 6-14 Aryl, 5 to 6 membered heteroaryl; for example, C 6-14 The aryl group is phenyl, and the 5- to 6-membered heteroaryl group is pyridin-3-yl or pyridin-4-yl;

[0077] Alternatively, the method comprises the following steps: in the presence of hydrogen and under the catalysis of catalyst 1, the compound shown in formula II is subjected to a reaction of removing the protecting group R' and reducing the olefin group in an organic solvent to obtain a compound shown in formula I, wherein R2' and R3' are acetyl groups or hydrogen atoms; wherein R1' is C 2-6 Alkenyl, for example vinyl, prop-1-en-2-yl, (E)-prop-1-en-1-yl or 2-methylprop-1-en-1-yl.

[0078] In some preferred embodiments of the present invention, the catalyst 1 is 20% palladium hydroxide on carbon; and / or the organic solvent is methanol.

[0079] In some embodiments of the present invention, the preparation method satisfies one or more of the following conditions:

[0080] (1) The molar volume ratio of the compound represented by formula II to the organic solvent is 0.002 mmol / mL to 0.025 mol / L, preferably 0.019 mmol / mL, 0.005 mmol / mL, 0.007 mmol / mL, 0.0075 mmol / mL or 0.003 mmol / mL;

[0081] (2) The molar mass ratio of the compound represented by formula II to the catalyst 1 is 0.0005 mmol / mg to 0.004 mmol / mg, preferably 0.0034 mmol / mg, 0.0029 mmol / mg, 0.0033 mmol / mg, 0.00125 mmol / mg or 0.00075 mmol / mg;

[0082] (4) The molar mass ratio of the compound represented by formula IV to the catalyst 1 is 0.0005 mmol / mg to 0.004 mmol / mg, preferably 0.0034 mmol / mg.

[0083] In some embodiments of the present invention, the preparation method satisfies one or more of the following conditions:

[0084] (1) The molar volume ratio of the compound represented by formula II to the organic solvent is 0.003 mmol / mL to 0.025 mol / L, preferably 0.019 mmol / mL, 0.005 mmol / mL or 0.007 mmol / mL;

[0085] (2) The molar mass ratio of the compound represented by formula II to the catalyst 1 is 0.002 mmol / mg to 0.004 mmol / mg, preferably 0.0034 mmol / mg, 0.0029 mmol / mg or 0.0033 mmol / mg.

[0086] In order to solve the above technical problems, the sixth aspect of the present invention provides a method for preparing a compound as shown in formula XI (for example, a compound as shown in formula II) as described in the second aspect of the present invention, wherein the method is any of the following methods:

[0087] Method 1: subjecting the above compound III to a reaction of removing the hydroxyl protecting group R";

[0088] Method 2: In a mixed solvent 2, under the protection of an inert gas, in the presence of a base 2, and under the catalysis of a catalyst 2, a compound as shown in formula IV and an R1' donor are subjected to a Suzuki coupling reaction;

[0089] Method 3: In an organic solvent, in the presence of a base 3, in the presence of an acylating agent, and under the catalysis of a catalyst 3, an acylation reaction is performed on the compound represented by formula II;

[0090] In method 1, the R" is preferably acetyl.

[0091] In method 2, R1' is defined as above; the inert gas is nitrogen or argon; and the Suzuki coupling reaction is carried out under heating reflux.

[0092] The compound represented by formula IV is preferably a compound represented by formula IV-1;

[0093]

[0094] Wherein, the definition of R' is the same as described above.

[0095] In some preferred embodiments of the present invention, the compound shown in formula IV is and / or, the R1' donor is potassium ethylene trifluoroborate, 4-pyridine boronic acid or 3-pyridine boronic acid; and / or, the catalyst 2 is palladium tetrakistriphenylphosphine; and / or, the mixed solvent 1 is dioxane and water, preferably in a volume ratio of 4:1; and / or, the base 1 is potassium carbonate.

[0096] In some preferred embodiments of the present invention, the R1' donor is methylboric acid, potassium ethylene trifluoroborate, isopropenylboronic acid pinacol ester, trans-1-propylene-1-boric acid, isobutyleneboronic acid pinacol ester, phenylboric acid, 4-pyridineboric acid or 3-pyridineboric acid; and / or, the catalyst 2 is tetrakistriphenylphosphine palladium; and / or, the mixed solvent 2 is selected from two or three of dioxane, water and acetonitrile, preferably dioxane and water, the volume ratio is 4:1, or dioxane, acetonitrile and water, the volume ratio is 3:3:1; and / or, the base 2 is potassium carbonate.

[0097] In method three, R2' and R3' are hydrogen atoms; and / or, the organic solvent is preferably dichloromethane; and / or, the base 3 is preferably triethylamine; and / or, the acylating agent is preferably acetic anhydride; and / or, the catalyst 3 is 4-dimethylaminopyridine.

[0098] More preferably, in method 1, the compound III is prepared by the following method:

[0099] In the mixed solvent 2, under the protection of the inert gas, in the presence of the base 2, and under the catalysis of the catalyst 2, the compound represented by formula V and the R1' donor are subjected to a Suzuki coupling reaction;

[0100]

[0101] Wherein, R' and R" are as defined above.

[0102] R1' is defined as above; the inert gas is nitrogen or argon; the Suzuki coupling reaction is carried out under heating reflux.

[0103] In some further more preferred embodiments of the present invention, the compound as shown in Formula V is and / or, the R1' donor is potassium ethylene trifluoroborate; and / or, the catalyst 2 is palladium tetrakistriphenylphosphine; and / or, the mixed solvent 2 is dioxane and water, preferably in a volume ratio of 4:1; and / or, the base 2 is potassium carbonate.

[0104] In some embodiments of the present invention, the preparation method satisfies one or more of the following conditions:

[0105] (1) The molar ratio of the compound represented by formula IV or V to the R1' donor is 1:1 to 1:1.5, preferably 1:1.29 or 1:1.3;

[0106] (2) The molar ratio of the compound represented by formula IV or V to the base 2 is 1:3 to 1:7, preferably 1:6.79, 1:3.14 or 1:6.83;

[0107] (3) The molar ratio of the compound represented by formula IV or V to the catalyst 2 is 1:0.1 to 1:0.15, preferably 1:0.12 or 1:0.13;

[0108] (4) The molar volume ratio of the compound represented by formula IV or V to the composite solvent 1 is 0.01 mmol / mL to 0.09 mmol / L, preferably 0.073 mmol / mL, 0.07 mmol / mL or 0.015 mmol / mL;

[0109] (6) The molar volume ratio of the compound represented by formula IV or V to the composite solvent 1 is 0.03 mmol / mL to 0.05 mmol / L, preferably 0.042 mmol / mL;

[0110] (7) The heating temperature is 60°C to 90°C, preferably 62°C or 87°C.

[0111] In some embodiments of the present invention, the above method 1 further comprises the following steps: in the presence of a base 4, heating the compound of formula 5 in a mixed solvent 3 under reflux to generate a compound of formula 3a;

[0112]

[0113] In some preferred embodiments of the present invention, the base 4 is sodium hydroxide; the molar ratio of the compound shown in Formula 5 to the base 4 is 1:7 to 1:9, preferably 1:8; the mixed solvent 3 is tetrahydrofuran and methanol, and the volume ratio is preferably 2:3.

[0114] In order to solve the above technical problems, the seventh aspect of the present invention provides a use of a polyphenol compound represented by formula X, its stereoisomers, and its pharmaceutically acceptable salts in the preparation of a drug for treating ischemic stroke;

[0115]

[0116] Where R1 is C 1-6 Alkyl, C 6-14 Aryl, 5- to 6-membered heteroaryl or H; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0117] R2 and R3 are independently a hydroxyl protecting group or hydrogen; the hydroxyl protecting group is C 1-6 Acyl.

[0118] Preferably, the above application is the use of the compound as described in the first aspect of the present invention, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in any scheme of the present invention in the preparation of a drug for treating ischemic stroke.

[0119] In some embodiments of the present invention, the medicament further comprises a pharmaceutically acceptable carrier.

[0120] In some embodiments of the present invention, when R1 is H, R2 and / or R3 are the hydroxyl protecting groups; for example, the polyphenol compound represented by formula X is

[0121] In some embodiments of the present invention, the manifestations of ischemic stroke are one or more of the following:

[0122] (1) behavioral abnormalities;

[0123] (2) cerebral infarction;

[0124] (3) HT22 hippocampal neuron cell damage.

[0125] In some embodiments of the present invention, the HT22 hippocampal neuron cell damage is manifested as: one or more of increased inflammatory factors, increased apoptotic factors and cell inactivation; the inflammatory factors are preferably iNOS and / or COX-2; the apoptotic factors are preferably one or more of p53, cleaved-Caspase3 and Bax; the cell inactivation is preferably reduced VCAM-1 protein expression and / or reduced cell viability detected by CCK-8 method.

[0126] In order to solve the above technical problems, the eighth aspect of the present invention provides a pharmaceutical composition, which comprises the polyphenol compound represented by formula X as described in the seventh aspect of the present invention, its stereoisomers, its pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0127] Preferably, the pharmaceutical composition comprises the compound as described in the first aspect of the present invention, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0128] Explanation of terms

[0129] The term "pharmaceutically acceptable" means that salts, solvents, excipients, etc. are generally non-toxic, safe, and suitable for use by subjects. The "subject" is preferably a mammal, more preferably a human.

[0130] The term "pharmaceutically acceptable salt" refers to a salt prepared from a compound of the present invention, a drug or a pharmaceutical composition comprising the same, and a relatively nontoxic, pharmaceutically acceptable acid or base. When the compound of the present invention, a drug or a pharmaceutical composition comprising the same contains a relatively acidic functional group, a base addition salt can be obtained by contacting a neutral form of such drug with a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent.

[0131] The term "stereoisomer" includes, for example, configurational isomers (eg, enantiomers or diastereomers) or conformational isomers (preferably rotational isomers). The isomers are, for example, conformational isomers, preferably rotational isomers.

[0132] The term "aryl" refers to a radical having the specified number of carbon atoms (e.g. 6-14 ) is a substituent having an aromatic ring structure, examples of aryl units include phenyl, substituted phenyl, naphthyl. It is understood that in "aryl", the connection with the parent structure is through the aromatic ring.

[0133] The "plurality" in the term "one or more" may mean 2, 3, 4, 5, 6, 7, 8, 9 or more.

[0134] The compound of the present invention, the medicament or the pharmaceutical composition containing the same can be administered in a unit dosage form, and the administration route can be enteral or parenteral.

[0135] The dosage form for administration may be a liquid dosage form, a solid dosage form or a semisolid dosage form.

[0136] The medicine or pharmaceutical composition of the present invention can be prepared into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0137] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of the present invention or their pharmaceutically acceptable salts, solvates, hydrates or prodrugs with other chemical components, such as pharmaceutically acceptable carriers. The purpose of a pharmaceutical composition is to facilitate the process of administration to an animal.

[0138] "Pharmaceutically acceptable carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to organisms and does not interfere with the biological activity and properties of the administered compound.

[0139] The aforementioned pharmaceutical composition, in addition to the pharmaceutically acceptable carrier, may also include adjuvants commonly used in pharmacy.

[0140] The term "prevention" refers to methods of preventing diseases and treating diseases.

[0141] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0142] The terms "compound" and "pharmaceutically acceptable salt" may exist in a crystalline or amorphous form. The term "crystalline form" means that the ions or molecules therein are arranged in a strict periodic manner in three-dimensional space in a certain manner, and have a regularity of periodic repetition at a certain distance; due to the different periodic arrangements mentioned above, there may be multiple crystalline forms, i.e., polymorphism. The term "amorphous form" means that the ions or molecules therein are in a disorderly distribution state, i.e., there is no periodic arrangement between the ions and molecules.

[0143] The term "C 1-6 The term "alkyl" refers to a straight or branched saturated aliphatic hydrocarbon group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, etc.).

[0144] The term "C2-6 The term "alkenyl" includes straight-chain or branched ethylenically unsaturated groups (eg, vinyl, allyl, methallyl, etc.).

[0145] The term "heteroaryl" means a 5-20-membered (preferably 5-12-membered, further preferably 5-10-membered, further preferably 5-8-membered, most preferably 5-6-membered) conjugated ring system group having 2 to 15 carbon atoms and 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur in the ring. Unless otherwise specified in this specification, the heteroaryl group may be a monocyclic, bicyclic, tricyclic or more ring system (preferably monocyclic). In a certain embodiment, the term "heteroaryl" refers to an aromatic group containing heteroatoms, each ring having aromaticity; preferably, the heteroatom species are independently selected from N, O and S, and the number of heteroatoms is 1, 2 or 3 5-10-membered heteroaryl or 5-6-membered heteroaryl. Examples of heteroaryl include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, diazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl (for example, ), benzopyrazolyl, indolyl, furanyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indolizinyl, isoxazolyl, thiadiazolyl, isoindolyl, indazolyl, isoindazolyl, purinyl, quinolyl, isoquinolyl, naphthazinyl, naphthyridinyl, quinoxalinyl, pteridinyl, carbazolyl, carbolyl, phenanthridinyl, phenanthrolinyl, acridinyl, phenazinyl, isothiazolyl, benzothiazolyl, benzisothiazolyl, benzothienyl, oxatriazolyl, cinnolinyl, quinazolinyl, indolizinyl, o-phenanthroline, isoxazolyl, phenoxazinyl, phenothiazinyl, benzoxazolyl or benzisoxazolyl (preferably pyridinyl).

[0146] In the present invention, the term “include, include or contain” may mean that in addition to the components listed below, there are other components; or it may mean “consisting of…”, that is, only the components listed below are included without other components.

[0147] The term "acyl" refers to an alkyl-CO- group, wherein alkyl is as defined above.

[0148] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0149] The reagents and raw materials used in the present invention are commercially available.

[0150] The positive and progressive effects of the present invention are: the compounds provided in the present application have one or more of the following advantageous effects:

[0151] (1) It has a significant protective effect against cerebral ischemia;

[0152] (2) It can significantly inhibit the inflammatory and apoptotic responses of neurons and reduce the volume of cerebral infarction in mice with cerebral ischemia;

[0153] (3) The polarity is reduced, which increases the oral bioavailability and the rate of crossing the blood-brain barrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0154] Figure 1 The Western blot method was used to detect the expression of inflammation-related proteins in HT22 hippocampal neurons damaged by OGD / R.

[0155] Figure 2 This is the quantitative analysis result of the Western blot method to detect the expression of inflammation-related proteins in HT22 hippocampal neurons damaged by OGD / R.

[0156] Figure 3 The Western blot method was used to detect the expression of apoptosis-related proteins in HT22 hippocampal neurons injured by OGD / R.

[0157] Figure 4 This is the quantitative analysis result of the Western blot method to detect the expression of apoptosis-related proteins in HT22 hippocampal neurons damaged by OGD / R.

[0158] Figure 5 Is 1a has a protective effect against cerebral infarction in a mouse cerebral ischemia model.

[0159] Figure 6 The effect of 1a on infarct volume in the mouse cerebral ischemia model.

[0160] Figure 7 It is 1a that improves the behavioral symptoms of cerebral ischemia in mice induced by cerebral ischemia model.

[0161] Figure 8 1b and 1c have protective effects on cerebral infarction in a mouse cerebral ischemia model.

[0162] Fig. 9 The effects of 1b and 1c on infarct volume in the mouse cerebral ischemia model. DETAILED DESCRIPTION

[0163] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0164] Example 1 (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-8-ethyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1a)

[0165]

[0166] Compound 2 (2.24 g, 1.63 mmol), potassium ethylene trifluoroborate (283 mg, 2.11 mmol), and potassium carbonate (1.53 g, 11.07 mmol) were added to a reaction flask, and 22.4 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, and tetrakistriphenylphosphine palladium (226 mg, 0.20 mmol) was added under nitrogen protection, the temperature was raised to 87°C and refluxed, and stirred for 24 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1, V / V) to obtain 1.52 g (yield 71%) of a light yellow solid product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-vinyl-[4,8'-dibenzodihydropyran]-3,3'-diol (3a).

[0167] 1 H-NMR(400MHz,CDCl3,7:3mixture of rotational isomers,δppm):7.51–6.71(m,48H),6.24(s,0.7H),6.16(s,0.3H),6.15(s,0.7H),6.07(s,0.3H ),5.94(dd,J=18.0,2.9Hz,0.7H),5.78(dd,J=18.0,2.9Hz,0.3H),5.20–4.44(m,18H),4.37(d,J= 8.9Hz,0.3H),4.23(m,0.7H),4.10(m,0.3H),3.75–3.51(m,1.7H),3.18(dd,J=16.4,6.0Hz,0.3H) ,3.03(dd,J=16.2,5.6Hz,0.7H),2.62(dd,J=16.4,9.7Hz,0.3H),2.38(dd,J=16.2,9.2Hz,0.7H).

[0168] Compound 3a (485 mg, 0.37 mmol), methanol (19 mL), and 20% palladium hydroxide on carbon (109 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 4 hours in the dark. After the reaction was completed, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 4: 1, V / V) to obtain a light yellow solid product (2R, 2'R, 3S, 3'S, 4S)-2,2'-di(3,4-dihydroxyphenyl)-8-ethyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1a) 139 mg (yield 63%).

[0169] 1 H-NMR (400MHz, D2O, δppm): 6.93–6.82 (m, 2H), 6.75 (dd, J = 8.2, 2.0Hz, 1H), 6.71 (d, J =8.2Hz,1H),6.48(d,J=2.0Hz,1H),6.28(dd,J=8.2,2.0Hz,1H),5.98(s,1H),4.62(d ,J=7.1Hz,1H),4.40–4.30(m,2H),4.28(m,1H),3.98(m,1H),2.73(dd,J=16.2,5.3Hz ,1H),2.47(dd,J=16.2,7.6Hz,1H),2.20(m,1H),2.05(m,1H),0.64(t,J=7.4Hz,3H). 13 C-NMR (100MHz, D2O, δppm):155.64,154.31,153.50,153.37,152.32,152.22,144.90,144.49,144.21,131.95,131.50,120.98,120.13,116 .84,116.43,116.37,115.10,111.53,109.79,107.89,101.65,82.15,80.84,73.32,67.27,37.78,26.85,16.20,13.88.HRMS(ESI)Calcdfor C 32 H 31 O 12 [M+H] + 607.1816, found 607.1810.

[0170] Example 2 (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-(pyridin-4-yl)-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1b)

[0171]

[0172] Compound 2 (100 mg, 0.07 mmol), 4-pyridineboronic acid (11 mg, 0.09 mmol) and potassium carbonate (30 mg, 0.22 mmol) were added to a reaction flask, 1 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, tetrakistriphenylphosphine palladium (10 mg, 8.65 μmol) was added under nitrogen protection, and the temperature was raised to 87 ° C and stirred for 16 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1, V / V) to obtain 58 mg (yield 58%) of a light yellow oily product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-(pyridin-4-yl)-[4,8'-dibenzodihydropyran]-3,3'-diol (3b).

[0173] 1 H-NMR(400MHz,CDCl3,57:43mixture of rotational isomers, δppm):8.43–8.37(m,2H),7.52–6.68(m,47.14H),6.50(d,J=8.4Hz,0.43H ),6.32–6.20(m,2H),6.14(s,0.43H),5.25–4.38(m,18.43H),4.22(m,0.57H),4.04 (m,0.43H),3.80–3.68(m,1.57H),3.19(dd,J=16.6,5.3Hz,0.43H),3.07(dd,J=16. 2,5.2Hz,0.57H),2.65(dd,J=16.6,9.2Hz,0.43H),2.42(dd,J=16.2,9.3Hz,0.57H).

[0174] Compound 3b (73 mg, 0.05 mmol), methanol (10 mL), and 20% palladium hydroxide on carbon (17 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 4 hours in the dark. After the reaction, the mixture was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 4: 1, V / V) to obtain an orange-yellow solid product (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-(pyridin-4-yl)-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1b) 16 mg (yield 46%).

[0175] 1 H-NMR(400MHz,D2O,δppm):8.05(m,2H),6.75–6.61(m,4H),6.57(dd,J=8.2,2.1Hz,1H),6.51(d,J=8.2Hz,1H),6.48(d,J=2.1Hz,1H),6.34(dd,J =8.2,2.1Hz,1H),4.45(d,J=7.8Hz,1H),4.32–4.19(m,2H),4.09(m,1H), 3.83(m,1H),2.74(dd,J=15.8,5.2Hz,1H),2.36(dd,J=15.8,7.3Hz,1H). 13 C-NMR (100MHz, D2O, δppm): 156.27, 156.00, 155.53, 154.77, 153.98, 151. 69,145.00,144.31,144.22,143.77,133.30,132.89,132.02,131.63,123 .06,120.11,119.39,116.98,116.86,116.55,115.07,110.42,107.52,10 6.62,102.11,82.71,81.15,72.91,67.07,37.08,26.05.HRMS(ESI)Calcd for C 35 H 30 NO 12 [M+H] + 656.1768, found 656.1755.

[0176] Example 3 (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-(pyridin-3-yl)-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1c)

[0177]

[0178] Compound 2 (100 mg, 0.07 mmol), 3-pyridineboronic acid (11 mg, 0.09 mmol) and potassium carbonate (30 mg, 0.22 mmol) were added to a reaction flask, 1 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, tetrakistriphenylphosphine palladium (10 mg, 8.65 μmol) was added under nitrogen protection, and the temperature was raised to 87 ° C and stirred for 16 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1, V / V) to obtain 47 mg (yield 47%) of a light yellow oily product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-(pyridin-3-yl)-[4,8'-dibenzodihydropyran]-3,3'-diol (3c).

[0179] 1 H-NMR(400MHz,CDCl3,53:47mixture of rotational isomers,δppm):8.51–8.47(m,1H),8.32(dd,J=4.6,1.6Hz,0.53H),8.22(dd,J=4.8,1.5Hz,0.47H),7.51 –6.69(m,47.06H),6.48(d,J=8.4Hz,0.47H),6.29(s,0.47H),6.26–6.17(m,1.53H),6.12(s,0.47H),5.2 2–4.37(m,18.47H),4.21(m,0.53H),4.03(m,0.47H),3.74–3.61(m,1.53H),3.18(dd,J=16.3,5.8Hz,0.4 7H), 3.04 (dd, J=16.6, 5.3Hz, 0.53H), 2.63 (dd, J=16.3, 9.2Hz, 0.47H), 2.41 (dd, J=16.6, 9.1Hz, 0.53H).

[0180] Compound 3c (92 mg, 0.07 mmol), methanol (10 mL), and 20% palladium hydroxide on carbon (21 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 4 hours in the dark. After the reaction was completed, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 4: 1, V / V) to obtain an orange-yellow solid product (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-(pyridin-3-yl)-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1c) 19 mg (yield 43%).

[0181] 1 H-NMR(400MHz,D2O,δppm):8.03(m,1H),7.67(m,1H),7.14–7.00(m,2H),6.70–6 .62(m,2H),6.57(dd,J=8.4,2.0Hz,1H),6.50(d,J=8.4Hz,1H),6.43(d,J=2.0Hz, 1H),6.31(dd,J=8.4,2.0Hz,1H),4.46(d,J=7.3Hz,1H),4.34–4.22(m,2H),4.09 (m,1H),3.82(m,1H),2.71(dd,J=16.1,5.3Hz,1H),2.35(dd,J=16.1,8.4Hz,1H). 13 C-NMR(100MHz,D2O,δppm):155.96,154.65,153.90,152.95,152.71,151.93, 149.86,149.83,145.33,144.37,144.14,143.73,133.48,132.92,132.00,131 .67,123.41,120.08,119.77,115.97,115.87,115.24,114.81,108.89,107.0 9,105.60,100.98,81.49,80.45,73.09,67.19,37.04,26.67.HRMS(ESI)Calcd for C 35 H 30 NO 12 [M+H] + 656.1768, found 656.1756.

[0182] Example 4 (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-8-methyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1d)

[0183]

[0184] Compound 2 (400 mg, 0.29 mmol), methylboric acid (26 mg, 0.43 mmol) and potassium carbonate (120 mg, 0.87 mmol) were added to a reaction flask, and 7 mL of a mixed solvent of dioxane: acetonitrile: water = 3:3:1 (V / V / V) was added to dissolve. Tetrakistriphenylphosphine palladium (34 mg, 0.03 mmol) was added under nitrogen protection, the temperature was raised to reflux, and stirring was carried out for 42 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1, V / V) to obtain 281 mg (yield 74%) of a light yellow solid product (2R,2'R,3S,3'S,4S)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-methyl-[4,8'-dibenzodihydropyran]-3,3'-diol (3d).

[0185] 1 H-NMR(400MHz,CDCl3,2:1mixture of rotational isomers,δppm):7.52–6.61(m,46H),6.25(s,0.67H),6.19(s,0.33H),6.14(s,0.67H), 6.07(s,0.33H),5.28–4.32(m,18.33H),4.22(m,0.67H),4.05(m,0.33H),3.75–3.52(m, 1.67H), 3.19 (dd, J=16.4, 6.1Hz, 0.33H), 3.03 (dd, J=16.2, 5.4Hz, 0.67H), 2.61 (dd, J= 16.4, 9.8Hz, 0.33H), 2.39 (dd, J=16.2, 8.8Hz, 0.67H), 1.97 (s, 2.01H), 1.85 (s, 0.99H).

[0186] Compound 3d (200 mg, 0.15 mmol), methanol (20 mL), and 20% palladium hydroxide on carbon (120 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 6 hours in the dark. After the reaction was completed, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 1: 1, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-8-methyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1d) 7 mg (yield 8%).

[0187] 1H-NMR (400MHz, D2O, δppm): 6.92–6.83 (m, 2H), 6.79 (d, J = 8.2Hz, 1H), 6.69 (dd, J=8.2,2.1Hz,1H),6.55(d,J=2.1Hz,1H),6.41(dd,J=8.2,2.1Hz,1H),6.13(s, 1H),6.00(s,1H),4.51(d,J=7.7Hz,1H),4.41–4.28(m,2H),4.21(m,1H),3.91( m,1H),2.86(dd,J=15.9,5.1Hz,1H),2.46(dd,J=15.9,8.3Hz,1H),1.60(s,3H). 13 C-NMR (100MHz, D2O, δppm): 154.97, 153.72, 153.05, 152.63, 151.98, 151.60, 144.24, 143.75, 143.71, 143.52, 131.09, 130.64, 120.28, 119. 91,116.16,115.78,114.85,109.27,107.07,104.03,101.32,96.16,95.28,81.44,80.38,72.68,66.88,37.08,27.32,7.09.HRMS(ESI)Calcd for C 31 H 29 O 12 [M+H] + 593.1659, found 593.1647.

[0188] Example 5 (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-isopropyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1e)

[0189]

[0190] Compound 2 (400 mg, 0.29 mmol), isopropenylboronic acid pinacol ester (63 mg, 0.37 mmol), and potassium carbonate (273 mg, 1.98 mmol) were added to a reaction flask, and 5 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, and tetrakistriphenylphosphine palladium (40 mg, 0.03 mmol) was added under nitrogen protection, the temperature was raised to reflux, and stirred for 40 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1, V / V) to obtain 323 mg of a light yellow solid product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-(prop-1-en-2-yl)-[4,8'-dibenzodihydropyran]-3,3'-diol (3e) (yield 83%).

[0191] 1 H-NMR(400MHz,CDCl3,2:1mixture of rotational isomers,δppm):7.47–6.57(m,46H),6.18(s,0.67H),6.17(s,0.33H),6.1 5(s,0.67H),6.09(s,0.33H),5.21–4.43(m,20.33H),4.25(m,0.67H),4.0 5(m,0.33H),3.76–3.59(m,1.67H),3.09–2.95(m,1H),2.64(dd,J=16.1,8 .0Hz, 0.33H), 2.36 (dd, J=16.4, 9.0Hz, 0.67H), 1.89 (s, 2H), 1.76 (s, 1H).

[0192] Compound 3d (200 mg, 0.15 mmol), methanol (20 mL), and 20% palladium hydroxide on carbon (120 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 2 hours in the dark. After the reaction, the mixture was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 1: 1, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4R)-2,2'-di(3,4-dihydroxyphenyl)-8-isopropyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1e) 28 mg (yield 30%).

[0193] 1H-NMR (400MHz, D2O, δppm): 6.97 (d, J = 2.0Hz, 1H), 6.90 (d, J = 8.2Hz, 1H), 6.84 (dd,J=8.2,2.0Hz,1H),6.66(d,J=8.2Hz,1H),6.42(d,J=2.0Hz,1H),6.16–6. 07(m,2H),5.99(s,1H),4.73(d,J=7.0Hz,1H),4.46–4.31(m,3H),4.09(m,1H) ,2.96(m,1H),2.62–2.42(m,2H),0.87(d,J=7.3Hz,3H),0.77(d,J=7.3Hz,3H). 13 C-NMR(100MHz,D2O,δppm):156.16,153.49,152.80,152.57,151.62,151.4 0,144.00,143.78,143.73,143.33,131.53,131.11,120.30,118.73,116.00 ,115.76,115.53,114.61,113.91,108.87,107.68,100.50,95.15,81.50,7 9.84,72.74,66.34,37.24,24.72,23.75,20.12,19.84.HRMS(ESI)Calcdfor C 33 H 33 O 12 [M+H] + 621.1972, found 621.1962.

[0194] Example 6 (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-8-propyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1f)

[0195]

[0196] Compound 2 (400 mg, 0.29 mmol), trans-1-propylene-1-boronic acid (37 mg, 0.43 mmol) and potassium carbonate (120 mg, 0.87 mmol) were added to a reaction flask, 5 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, tetrakistriphenylphosphine palladium (40 mg, 0.03 mmol) was added under nitrogen protection, the temperature was raised to reflux, and stirring was carried out for 42 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1, V / V) to obtain 262 mg of a light yellow solid product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-((E)-prop-1-en-1-yl)-[4,8'-dibenzodihydropyran]-3,3'-diol (3f) (yield 67%).

[0197] 1 H-NMR(400MHz,CDCl3,2:1mixture of rotational isomers,δppm):7.51–6.27(m,48H),6.23(s,0.67H),6.16(s,0.33H),6.14(s,0.67H),6.07(s, 0.33H),5.22–4.44(m,18H),4.37(d,J=9.0Hz,0.33H),4.21(m,0.67H),4.09(m,0.33H),3.72–3. 52(m,1.67H),3.18(dd,J=16.5,6.3Hz,0.33H),3.03(dd,J=16.5,5.7Hz,0.67H),2.62(dd,J=16. 5,9.9Hz,0.33H),2.37(dd,J=16.5,9.1Hz,0.67H),1.81(d,J=6.5Hz,2H),1.73(d,J=6.5Hz,1H).

[0198] Compound 3f (200 mg, 0.15 mmol), methanol (20 mL), and 20% palladium hydroxide on carbon (120 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 2 hours in the dark. After the reaction was completed, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 1: 1, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4S)-2,2'-di(3,4-dihydroxyphenyl)-8-propyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1f) 11 mg (yield 12%).

[0199] 1 H-NMR (400MHz, D2O, δppm): 6.91 (d, J = 2.1Hz, 1H), 6.89 (d, J = 8.1Hz, 1H), 6.76 (dd, J = 8.2, 2. 2Hz,1H),6.71(d,J=8.2Hz,1H),6.49(d,J=2.2Hz,1H),6.27(dd,J=8.1,2.1Hz,1H),6.13(s,1 H),6.00(s,1H),4.66(d,J=7.0Hz,1H),4.41–4.23(m,3H),4.01(m,1H),2.71(dd,J=16.3,5.3 Hz,1H),2.48(dd,J=16.3,7.4Hz,1H),2.23–1.98(m,2H),1.07(m,2H),0.61(t,J=7.4Hz,3H). 13 C-NMR (100MHz, D2O, δppm): 155.24, 153.64, 152.79, 152.71, 151.88, 151. 67,144.16,143.76,143.53,143.51,131.29,130.81,120.33,119.37,116. 12,115.70,115.68,114.35,109.37,109.06,107.09,100.87,96.33,81.44 ,80.10,72.66,66.51,37.18,25.93,24.04,22.26,13.07.HRMS(ESI)Calcd for C 33 H 33 O 12 [M+H] + 621.1972, found 621.1960.

[0200] Example 7 (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-8-isobutyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1 g)

[0201]

[0202] Compound 2 (400 mg, 0.29 mmol), isobutylene boronic acid pinacol ester (69 mg, 0.38 mmol), and potassium carbonate (273 mg, 1.98 mmol) were added to a reaction flask, and 5 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, and tetrakistriphenylphosphine palladium (40 mg, 0.03 mmol) was added under nitrogen protection, the temperature was raised to reflux, and stirred for 40 hours. After the reaction, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1, V / V) to give a light yellow solid product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-(2-methylprop-1-en-1-yl)-[4,8'-dibenzodihydropyran]-3,3'-diol (3 g) 377 mg (yield 96%).

[0203] 1 H-NMR(400MHz,CDCl3,7:3mixture of rotational isomers,δppm):7.52–6.61(m,46H),6.20–6.03(m,2H),5.94(s,0.7H),5.81(s,0. 3H),5.23–4.41(m,18.3H),4.24(m,0.7H),4.09(m,0.3H),3.76–3.60(m,1H),3.56 (d,J=8.6Hz,0.7H),3.07–2.96(m,1H),2.65(dd,J=16.8,8.7Hz,0.3H),2.35(dd,J =16.2,9.5Hz,0.7H),1.99(s,0.9H),1.81(s,2.1H),1.71(s,0.9H),1.43(s,2.1H).

[0204] Compound 3g (200 mg, 0.15 mmol), methanol (20 mL), and 20% palladium hydroxide on carbon (120 mg) were added to the reaction flask, replaced with hydrogen, and stirred for 2 hours in the dark. After the reaction, the mixture was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 2: 3, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4S)-2,2'-di(3,4-dihydroxyphenyl)-8-isobutyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1 g) 23 mg (yield 24%).

[0205] 1H-NMR(400MHz,CD3OD,3:2mixture of rotational isomers,δppm):6.98–6.77(m,2H),6.75(dd,J=8.1,1.5Hz,0.6H),6.68(d,J=8.2Hz,0.6H),6.62–6.55(m,1.6H),6.47(d,J=2.2Hz,0.6H),6.11(dd,J=8.2,2.2Hz,0.6H),6.05(s,0.6H),5.93–5.89(m,1H),5.84(s,0.4H),4.74(d,J=7.2Hz,0.4H),4.63(d,J=6.3Hz,0.6H),4.50(d,J=7.8Hz,0.4H),4.46–4.33(m,1.6H),4.27(m,0.4H),4.20(m,0.6H),4.07(m,0.4H),3.87(m,0.6H),2.79(dd,J=16.3,5.5Hz,0.4H),2.66–2.16(m,3.6H),1.79(m,0.4H),1.59(m,0.6H),0.85(d,J=6.6Hz,1.2H),0.79(d,J=6.6Hz,1.2H),0.69(d,J=6.6Hz,1.8H),0.60(d,J=6.6Hz,1.8H). 13C-NMR(100MHz,CD3OD,3:2mixture of rotational isomers, δppm):156.86,155.87,155.85,155.70,155.42,155.22,155.01,154 .68,154.60,154.35,153.99,146.11,146.05,145.91,145.76,145.51,145.34 ,133.14,132.93,132.05,131.90,120.76,120.56,120.11,119.32,116.39,116.16,116.03,115.86,115.79,115.14,114.90,109.61,109.09,108.72,108.20 106.90,106.62,101.92,100.38,97.59,97.26,97.00,95.86,83.80,83.60,82.94,81.97,74.37,73.86 ,68.46,38.93,38.74,32.66,29.94,29.91,28.33,27.53,23.11,23.01,22.99,22.76.HRMS(ESI)Calcd for C 34 H 35 O 12 [M+H] + 635.2129, found 635.2120.

[0206] Example 8 (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-8-phenyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1h)

[0207]

[0208] Compound 2 (400 mg, 0.29 mmol), phenylboric acid (53 mg, 0.43 mmol), potassium carbonate (120 mg, 0.87 mmol) were added to a reaction flask, and a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve 5 mL, and tetrakistriphenylphosphine palladium (34 mg, 0.03 mmol) was added under nitrogen protection, and the temperature was raised to reflux and stirred for 42 hours. After the reaction was completed, the organic phase was concentrated in vacuo after the filtrate was separated, and then purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1, V / V) to obtain a light yellow solid product (2R, 2'R, 3S, 3'S, 4R)-5,5',7,7'-tetra (benzyloxy)-2,2'-di (3,4-di (benzyloxy) phenyl)-8-phenyl-[4,8'-dibenzodihydropyran]-3,3'-diol (3h) 336 mg (yield 84%).

[0209] 1 H-NMR(400MHz,CDCl3,53:47mixture of rotational isomers,δppm):7.55–6.47(m,51H),6.30(s,0.47H),6.25(s,0.53H),6.22(s, 0.53H),6.11(s,0.47H),5.23–4.36(m,18.47H),4.20(m,0.53H),4.01(m,0.47H ),3.77–3.55(m,1.53H),3.19(dd,J=16.4,5.8Hz,0.47H),3.04(dd,J=16.3,5. 7Hz, 0.53H), 2.63 (dd, J=16.4, 9.7Hz, 0.47H), 2.38 (dd, J=16.3, 9.1Hz, 0.53H).

[0210] Compound 3h (80 mg, 0.06 mmol), methanol (20 mL), and 20% palladium hydroxide on carbon (80 mg) were added to the reaction flask, replaced with hydrogen, and stirred for 4 hours in the dark. After the reaction was completed, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 2: 3, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4R)-2,2'-di(3,4-dihydroxyphenyl)-8-phenyl-[4,8'-dibenzodihydropyran]-3,3',5,5',7,7'-hexanol (1h) 8 mg (yield 21%).

[0211] 1H-NMR (400MHz, D2O, δppm): 7.26–7.12(m,3H),6.82–6.70(m,5H),6.63(d,J=8.0Hz,1H),6.60(d,J=2.0Hz,1H),6.39(dd,J=8.0,2.0Hz ,1H),6.11(s,1H),4.62(d,J=7.2Hz,1H),4.47–4.26(m,3H),4.01(m,1H),2.78(dd,J=16.0,5.4Hz,1H),2.48(dd,J=16.0,7.9Hz,1H). 13 C-NMR(100MHz,D2O,δppm):153.77,153.64,152.92,152.87,151.69,151.50, 144.31,143.85,143.67,143.30,133.31,131.04,130.88,130.64,127.81,127 .16,126.72,119.97,119.39,115.91,115.85,115.36,114.79,109.78,108.7 8,106.94,100.82,81.53,80.32,72.36,66.82,37.03,26.26.HRMS(ESI)Calcd for C 36 H 31 O 12 [M+H] + 655.1816, found 655.1804.

[0212] Example 9 (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-5,5',7,7'-tetrahydroxy-8-isopropyl-[4,8'-dibenzodihydropyran]-3,3'-diyl diacetate (1i)

[0213]

[0214] Compound 3e (811 mg, 0.61 mmol), dichloromethane (5 mL), DMAP (7.4 mg, 0.06 mmol), triethylamine (307 mg, 3.03 mmol) were added to the reaction flask, acetic anhydride (148 mg, 1.45 mmol) was added to the reaction system, and stirred at room temperature for 3 hours. After the reaction was completed, water was added for washing, 10% hydrochloric acid aqueous solution was added to acidify the aqueous phase to pH = 6, and then saturated sodium chloride aqueous solution was washed, the organic phase was added to anhydrous sodium sulfate for drying, and vacuum concentrated to obtain a yellow oily product (2R, 2'R, 3S, 3'S, 4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-(prop-1-en-2-yl)-[4,8'-dibenzodihydropyran]-3,3'-diyl diacetate (3i), which was directly used in the next step.

[0215] Methanol (40 mL) and 20% palladium hydroxide on carbon (280 mg) were added to the reaction bottle of compound 3i, and hydrogen was replaced and stirred for 4 hours in the dark. After the reaction, it was filtered with a microporous filter membrane (nylon 6, 0.45 μm), and the filtrate was concentrated in vacuo and purified by ODS column chromatography (water: methanol = 2: 3, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4R)-2,2'-bis(3,4-dihydroxyphenyl)-5,5',7,7'-tetrahydroxy-8-isopropyl-[4,8'-dibenzodihydropyran]-3,3'-diyl diacetate (1i) 108 mg (total yield of two steps 25%).

[0216] 1 H-NMR (400MHz, D2O, δppm): 6.99 (d, J = 1.8 Hz, 1H), 6.88 ( d, J = 8.2 Hz, 1H), 6.84 ( dd, J = 8.2, 1.8 Hz, 1H), 6.66 ( d,J=8.2Hz,1H),6.42(d,J=1.8Hz,1H),6.09(s,1H),6.08(s,1H),5.99(dd,J=8.2,1.8Hz,1H),5.66(m,1H),5 .31(m,1H),5.25(m,1H),4.59(d,J=7.5Hz,1H),4.52(d,J=10.1Hz,1H),3.03(m,1H),2.65(dd,J=16.0,3.0H z,1H),2.47(dd,J=16.0,4.2Hz,1H),1.98(s,3H),1.83(s,3H),0.93(d,J=7.0Hz,3H),0.85(d,J=7.0Hz,3H). 13C-NMR (100MHz, D2O, δppm):173.78,172.73,156.16,155.01,154.15,153.63,153.2 0,151.79,144.12,143.83,143.66,143.43,130.78,130.30,119.54,118.07,115.89 ,115.75,114.94,114.80,113.18,108.24,107.07,99.49,97.06,95.09,78.68,76. 28,75.15,69.38,34.64,23.81,21.09,20.41,20.09,20.05,19.98.HRMS(ESI)Calcd for C 37 H 37 O 14 [M+H] + 705.2183, found 705.2180.

[0217] Example 10 (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-5,5',7,7'-tetrahydroxy-[4,8'-dibenzodihydropyran]-3,3'-diyl diacetate (1j)

[0218]

[0219] Compound 2a (20 mg, 13.69 μmol), 3 mL of a mixed solvent of tetrahydrofuran:methanol = 1:2 (V / V), sodium bicarbonate (3.5 mg, 41.66 μmol), and 20% palladium hydroxide on carbon (4 mg) were added to a reaction flask, replaced with hydrogen, and stirred for 6 hours in the dark. After the reaction, acetic acid (1.7 mg, 28.31 μmol) was added, stirred for 10 minutes, and filtered with a microporous filter membrane (nylon 6, 0.45 μm). The filtrate was concentrated in vacuo and purified by ODS column chromatography (water:methanol = 2:3, V / V) to obtain a yellow solid product (2R, 2'R, 3S, 3'S, 4S)-2,2'-bis(3,4-dihydroxyphenyl)-5,5',7,7'-tetrahydroxy-[4,8'-dibenzodihydropyran]-3,3'-diyl diacetate (1j) 7 mg (yield 77%).

[0220] 1H-NMR (400MHz, D2O, δppm): 6.88–6.70 (m, 3H), 6.65–6.52 (m, 2H), 6.29 (dd, J = 7.3, 2.1Hz,1H),6.10(s,1H),6.04(d,J=2.6Hz,1H),5.80(d,J=2.6Hz,1H),5.69(m,1H), 5.15(m,1H),4.98(d,J=6.5Hz,1H),4.57(d,J=9.9Hz,1H),4.53(d,J=8.3Hz,1H),2. 73(dd,J=16.5,5.4Hz,1H),2.63(dd,J=16.5,6.7Hz,1H),1.95(s,3H),1.73(s,3H). 13 C-NMR (100MHz, D2O, δppm): 173.36, 172.56, 156.32, 155.06, 153.49, 153. 09,152.65,144.50,143.79,143.65,130.00,129.40,120.00,118.97,115. 94,115.60,115.40,114.11,107.73,106.17,100.14,97.00,95.44,95.30 ,79.21,77.18,73.97,69.60,34.53,23.31,20.36,19.83.HRMS(ESI)Calcd for C 34 H 31 O 14 [M+H] + 663.1714, found 663.1704.

[0221] Example 11 (2R, 2'R, 3S, 3'S, 4R)-5,5',7,7'-tetrakis(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-vinyl-[4,8'-dibenzodihydropyran]-3,3'-diol (3a)

[0222]

[0223] Compound 4 (429 mg, 0.30 mmol), potassium ethylene trifluoroborate (53 mg, 0.39 mmol) and potassium carbonate (284 mg, 2.05 mmol) were added to a reaction flask, 20 mL of a mixed solvent of dioxane: water = 4:1 (V / V) was added to dissolve, tetrakistriphenylphosphine palladium (42 mg, 0.04 mmol) was added under nitrogen protection, the temperature was raised to 87 ° C and refluxed, and stirred for 18 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was separated, and the organic phase was concentrated in vacuo. The mixture was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1, V / V) to obtain a light yellow solid product (2R,2'R,3S,3'S,4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-3'-hydroxy-8-vinyl-[4,8'-dibenzodihydropyran]-3-yl acetate (5), 286 mg (yield 69%). 1 H-NMR(400MHz,CDCl3,1:1mixture of rotational isomers, δppm): 7.50–6.61 (m, 47H), 6.39 (dd, J = 8.2, 1.5Hz, 0.5H), 6.23 (s, 0.5H), 6.20 (s, 0.5H), 6.1 8(s,0.5H),6.13–6.07(m,1H),5.97–5.94(m,1H),5.91–5.80(m,1H),5.20–4.48(m,18.5H),3.90(m,0. 5H),3.54(m,0.5H),3.30(d,J=8.8Hz,0.5H),3.02(dd,J=16.7,6.0Hz,0.5H),2.82(dd,J=16.1,6.5Hz, 0.5H), 2.71 (dd, J=16.1, 7.9Hz, 0.5H), 2.32 (dd, J=16.7, 8.1Hz, 0.5H), 1.60 (s, 1.5H), 1.51 (s, 1.5H).

[0224] Compound 5 (286 mg, 0.21 mmol), tetrahydrofuran (2 mL), and methanol (3 mL) were added to a reaction flask, and sodium hydroxide (67 mg, 1.68 mmol) was slowly added, and the temperature was raised to 62°C for reflux and stirred for 2 hours. After the reaction was completed, the mixture was concentrated, ethyl acetate (5 mL) was added for dissolution, and the mixture was washed once with a saturated sodium chloride aqueous solution (5 mL). The organic phase was concentrated in vacuo and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1, V / V) to obtain a light yellow solid product (2R, 2'R, 3S, 3'S, 4R)-5,5',7,7'-tetra(benzyloxy)-2,2'-di(3,4-di(benzyloxy)phenyl)-8-vinyl-[4,8'-dibenzodihydropyran]-3,3'-diol (3a), 196 mg (yield 71%).

[0225] Example 12 Protective effects of 1a, 1b and 1c on HT22 hippocampal neuronal cells injured by OGD / R (oxygen glucose deprivation reperfusion) in vitro

[0226] Grouping: HT22 hippocampal neuron cells (purchased from Thermo Fisher Scientific) were cultured in a medium: 10% fetal bovine serum (purchased from Sigma) + 1% penicillin-streptomycin double antibody solution (Shanghai Yuanpei Biotechnology Co., Ltd.) + 89% DMEM / F12 medium (Gibco), and randomly divided into 6 groups, namely: 1) Control group; 2) OGD / R group; 3) Proanthocyanidin B3 (referred to as B3) (20μM) group; 4) 1a (20μM) group; 5) 1b (20μM) group; 6) 1c (20μM) group. All compounds to be tested were dissolved in DMSO and then diluted to the specified concentration with culture medium, and the final concentration of DMSO was 0.1%. For groups 3) to 6), B3 / 1a / 1b / 1c was given 4h in advance (at a concentration of 20μM, respectively), and after stimulating the cells for 4h, they were deprived of oxygen and glucose for 2h, and then reoxygenated for 24h to prepare an in vitro cerebral ischemia model. For group 2), oxygen and glucose deprivation was followed by reoxygenation for 24 hours. Proteins were collected and Western blot was used to detect the expression of β-tubulin, p53, cleaved-Caspase3, cleaved-Caspase9, Bcl-2, iNOS, COX-2, and VCAM-1 proteins. The results showed (Table 1) that B3 / 1a / 1b / 1c could significantly reduce the expression of inflammatory proteins (see Figure 1 , Figure 2 ); at the same time, B3 / 1a / 1b / 1c can significantly reduce the expression of apoptotic proteins (cleaved-Caspase3, cleaved-Caspase9) (see Figure 3 , Figure 4 ), and 1a / 1b / 1c is better than B3.

[0227] Figure 1-4 middle, * Compared with the OGD / R group, * p<0.05, ** p<0.01, *** p<0.001.

[0228] The experimental results show that the compounds 1a / 1b / 1c have a significant protective effect on the in vitro cerebral ischemia model induced by OGD / R, and can be used to prepare drugs for treating cerebral ischemia-related diseases.

[0229] Table 1 Protective effects of 1a, 1b and 1c on HT22 hippocampal neurons injured by OGD / R (oxygen glucose deprivation reperfusion) in vitro

[0230]

[0231] Example 13 The therapeutic effect of 1a on the in vivo mouse cerebral ischemia model

[0232] Grouping: Male C57 mice weighing 21-24g (mice purchased from Shanghai B&K Company, SPF grade) were anesthetized by intraperitoneal injection of 1% sodium pentobarbital and fixed in supine position. The skin in the middle of the neck was opened with an incision of about 2cm. The subcutaneous tissue was bluntly separated to expose the left common carotid artery (CCA), and the external carotid artery (ECA) and internal carotid artery (ICA) were separated and exposed upwards in sequence. The common carotid artery and the distal end of the external carotid artery were hung with a thread, and the common carotid artery was ligated with a slipknot, and the external carotid artery was ligated with a dead knot. Then the thread was hung at the distal end of the internal carotid artery and pulled with a retractor. The thread was hung at the proximal end of the external carotid artery and a slipknot was tied. Then, at the "Y-shaped" bifurcation of the external carotid artery and the internal carotid artery, use microscissors to make a small cut at the eccentric external carotid artery, guide the thread plug from the incision into the external carotid artery, and then insert it into the common carotid artery. Then tighten the ligature, cut off one side of the external carotid artery, and slightly pull the stump of the external carotid artery so that the external carotid artery and the internal carotid artery are in a straight line. Adjust the angle of the thread plug to form a 45° horizontal angle with the middle of the front neck. After the thread plug is inserted into the internal carotid artery, remove the hanging line of the internal carotid artery, and gently guide the thread plug into the skull. Stop when resistance is encountered, and then fix the ligature thread plug. Cut off the excess thread plug, leaving only a stump of about 1mm long. Remove the hanging line of the common carotid artery, observe for bleeding, and suture the incision. After 1.5 hours of infarction, open the incision and pull out the thread plug to the branch of the external carotid artery and the common carotid artery. Suture the skin, place it in a cage after iodine disinfection, pay attention to keeping warm throughout the process, and provide sufficient water and food after surgery. The sham operation group was operated in the same way as the model group except that no suture was inserted. 1a was dissolved in ddH2O, and the mice were randomly divided into the model group (MCAO, solvent control, ip.n=4), the model + proanthocyanidin B3 group (48 mg / kg / day, ip.n=4), and the model + 1a group (48 mg / kg / day, ip.n=4). The drug was administered once within 6 hours after surgery, and then continuously for 3 days. 72 hours after modeling, behavioral evaluation was performed; TTC was used to detect cerebral infarction volume. The results showed that 1a could improve the behavioral symptoms of mice with cerebral ischemia and reduce the volume of cerebral infarction (see Table 2, Table 3, Figure 5 , Figure 6 , Figure 7 ).

[0233] Table 2 Protective effect of 1a on cerebral infarction in induced cerebral ischemia model in mice

[0234]

[0235] Table 3 Improvement of behavioral symptoms of cerebral ischemia mice by 1a in induced cerebral ischemia model

[0236]

[0237]

[0238] The experimental results demonstrate that the 1a has a significant protective effect on the in vivo cerebral ischemia model prepared by the thread embolization method.

[0239] In summary: The present invention adopts the C57 mouse embolization method to prepare a cerebral ischemic injury model, and the Bederson score is used to detect the behavioral damage of mice after cerebral ischemia. The results show that 1a can improve the behavioral symptoms of mice with cerebral ischemia; the TTC method is used to detect the volume of cerebral infarction after cerebral ischemia in mice, and the results show that 1a can reduce the volume of cerebral infarction. In addition, a cerebral ischemia model was established in vitro by using OGD / R-induced HT22 mouse hippocampal neuronal cells to explore the effect of 1a on neuronal cells after cerebral ischemia. Western blot results show that it can significantly inhibit the inflammatory and apoptotic responses of neurons. It is proved that the compound has a protective effect on cerebral ischemic diseases.

[0240] Example 14 Therapeutic effects of 1b and 1c on in vivo mouse cerebral ischemia model

[0241] Grouping: Male C57 mice weighing 21-24g (mice purchased from Shanghai B&K Company, SPF grade) were anesthetized by intraperitoneal injection of 1% sodium pentobarbital and fixed in supine position. The skin in the middle of the neck was opened with an incision of about 2cm. The subcutaneous tissue was bluntly separated to expose the left common carotid artery (CCA), and the external carotid artery (ECA) and internal carotid artery (ICA) were separated and exposed upwards in sequence. The common carotid artery and the distal end of the external carotid artery were hung with a thread, and the common carotid artery was ligated with a slipknot, and the external carotid artery was ligated with a dead knot. Then the thread was hung at the distal end of the internal carotid artery and pulled with a retractor. The thread was hung at the proximal end of the external carotid artery and a slipknot was tied. Then, at the "Y-shaped" bifurcation of the external carotid artery and the internal carotid artery, use microscissors to make a small cut at the eccentric external carotid artery, guide the thread plug from the incision into the external carotid artery, and then insert it into the common carotid artery. Then tighten the ligature, cut off one side of the external carotid artery, and slightly pull the stump of the external carotid artery so that the external carotid artery and the internal carotid artery are in a straight line. Adjust the angle of the thread plug to form a 45° horizontal angle with the middle of the front neck. After the thread plug is inserted into the internal carotid artery, remove the hanging line of the internal carotid artery, and gently guide the thread plug into the skull. Stop when resistance is encountered, and then fix the ligature thread plug. Cut off the excess thread plug, leaving only a stump of about 1mm long. Remove the hanging line of the common carotid artery, observe for bleeding, and suture the incision. After 1.5 hours of infarction, open the incision and pull out the thread plug to the branch of the external carotid artery and the common carotid artery. Suture the skin, place it in a cage after iodine disinfection, pay attention to keeping warm throughout the process, and provide sufficient water and food after surgery. The sham operation group was operated in the same way as the model group except that no suture was inserted. 1b / 1c was dissolved in 2% DMSO+30% PEG 300+5% Tween 80+ddH2O in sequence, and the mice were randomly divided into the model group (MCAO, solvent control, ip.n=3), model+1b group (40mg / kg / day, ip.n=4), and model+1c group (40mg / kg / day, ip.n=4). The drug was intraperitoneally administered once within 3 hours after surgery, and the samples were collected 24 hours after modeling, and the cerebral infarction volume was detected by TTC. The results showed that 1b / 1c could reduce the cerebral infarction volume of mice with cerebral ischemia (Table 4, Fig. 9 and Figure 8 ).

[0242] Table 4 Protective effects of 1b and 1c on cerebral infarction in induced cerebral ischemia model in mice

[0243]

[0244] The experimental results show that the 1b / 1c has a protective effect on the in vivo cerebral ischemia model prepared by the thread embolization method.

[0245] Example 15 Bioavailability Test of Compound 1a

[0246] 1) Experimental animals: SD rats, weighing about 200 g, were kept in a constant temperature, purified and ventilated animal room at 20-26°C, with free access to food and water, humidity at 40-70%, and light exposure for 12 h.

[0247] 2) Administration method: Route: single oral administration, volume: calculated as 10 mL / kg body weight; Route: single intravenous injection, volume: calculated as 5 mL / kg body weight.

[0248] 3) Administration and sample collection: 24 SD rats were divided into 4 groups, 6 rats in each group, half of them were male and half were female. Fasting for 12 hours before administration, free access to water. A single oral gavage of 50 mg / kg and a single intravenous injection of 2 mg / kg were respectively administered. For oral gavage, about 200 μL of blood was collected from the eye socket before administration and 2min, 5min, 10min, 15min, 0.5h, 1h, 2h, 4h, 6h, 9h, and 24h after administration; for intravenous administration, blood was collected from the eye socket before administration and 3min, 8min, 15min, 0.5h, 1h, 2h, 4h, 6h, 9h, and 24h after administration. The blood was anticoagulated with 1% heparin and centrifuged at 8000rpm for 5min to separate the plasma. It was stored at -80℃ for testing. High-concentration samples were diluted with blank plasma.

[0249] 4) Plasma sample determination method: Liquid chromatography system: ExionLC liquid chromatography system, Shimadzu Corporation; MS / MS system: 5500 tandem mass spectrometer equipped with electrospray ionization source (ESI source), AB Company; Data acquisition: Analyst (version 1.7), AB Company. Chromatographic column: HSS T3 (100×2.1 mm ID, 1.8 μm particle size), injection volume: 5 μL.

[0250] Mobile phase: A: methanol; C: aqueous solution containing 0.1% formic acid, flow rate: 0.35 mL / min.

[0251] 5) Sample processing: Take 50 μL of plasma sample, add 5 μL of 50% methanol water and 5 μL of internal standard (50 ng / mL tolbutamide), vortex mix for 10 seconds, add 150 μL of methanol containing 0.5% formic acid, and centrifuge at 14000 rpm for 6 minutes. The supernatant is filtered with a 0.22 μm filter head and analyzed by LC-MS / MS.

[0252] The results showed that the bioavailability of 50 mg / kg compound 1a in SD rats was 5.2%, which was significantly better than the 1.5% of proanthocyanidin B3.

[0253] Example 16 Brain tissue distribution experiment of compound 1a

[0254] 1) Experimental animals: SD rats, weighing about 200 g, were kept in a constant temperature, purified and ventilated animal room at 20-26°C, with free access to food and water, humidity at 40-70%, and light exposure for 12 h.

[0255] 2) Administration method: Route: single oral administration, volume: calculated as 10mL / kg body weight.

[0256] 3) Administration and sample collection: 18 male SD rats were divided into 2 groups. They were fasted for 12 hours before administration and had free access to water. A single oral administration of 50 mg / kg was performed. 5 min (3 rats), 0.5 h (3 rats), and 2 h (3 rats) after administration were performed, and brain tissues were obtained. Blood was collected at the same time and centrifuged to obtain plasma. The rats were frozen at -80°C until the concentration was determined.

[0257] 4) Tissue sample determination method: Liquid chromatography system: ExionLC liquid chromatography system, Shimadzu Corporation; MS / MS system: 5500 tandem mass spectrometer equipped with electrospray ionization source (ESI source), AB Company; Data acquisition: Analyst (version 1.7), AB Company. Chromatographic column: HSS T3 (100×2.1 mm ID, 1.8 μm particle size), injection volume: 5 μL.

[0258] Mobile phase: A: methanol; C: aqueous solution containing 0.1% formic acid, flow rate: 0.35 mL / min.

[0259] 5) Sample processing: Take 100 mg of tissue sample, add 10 μL 50% methanol water and 10 μL internal standard (10 ng / mL tolbutamide), and homogenize in a high-throughput tissue homogenizer. Use 500 μL methanol containing 0.5% formic acid to precipitate protein, and centrifuge at 14000 rpm for 6 min. Take 450 μL of supernatant and blow it with nitrogen (40°C), blow it dry, and then re-dissolve it with 100 μL 20% methanol water containing 0.1% formic acid, then ultrasonicate it in an ice bath for 5 min, and then centrifuge it at 14000 rpm for 6 min, and take 5 μL of supernatant for injection analysis.

[0260] The results showed that the concentration of 1a in the rat brain was 10.35 ng / g after 2 hours, which was significantly better than the 2.52 ng / g of proanthocyanidin B3.

[0261] Example 17 Effects of Compounds 1a-1j on the Viability of HT22 Hippocampal Neuronal Cells Injured by OGD / R (Oxygen Glucose Deprivation Reperfusion) in Vitro

[0262] Experimental subjects: HT22 hippocampal neuronal cells (purchased from Thermo Fisher Scientific), cultured with the following culture medium: 10% fetal bovine serum (purchased from Sigma) + 1% penicillin-streptomycin double antibody solution (Shanghai Yuanpei Biotechnology Co., Ltd.) + 89% DMEM / F12 culture medium (Gibco); control compound was EDA (purchased from Shanghai Titan Technology Co., Ltd.)

[0263] All compounds to be tested were dissolved in DMSO and then diluted with culture medium to the specified concentration. The final concentration of DMSO was 0.1%. The cells in the model group were first subjected to oxygen and glucose deprivation for 6 hours, and then treated with different compounds at different concentrations for 24 hours; an equal volume of culture medium containing 0.1% DMSO was added to the cells in the control group. Cell viability was determined using a cell counting kit-8 (CCK-8, purchased from Yishen Biotechnology (Shanghai) Co., Ltd.). CCK-8 (100 μL / mL) was added to each well, and an equal amount of solvent was added to the blank group, and incubated at 37°C in the dark for 1 hour. Then, the absorbance was read at 450nm using the BioTek Synergy H1 multi-function microplate reader to calculate cell viability. The data were analyzed using a single Factorial ANOVA, Dunnett's multiple comparison test Compared with the OGD / R group, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

[0264] Table 5 Effects of Compounds 1a-1h (20 μM) on the viability of HT22 hippocampal neurons injured by OGD / R (oxygen glucose deprivation reperfusion) in vitro

[0265] Compound Cell viability (% relative to the control group) 1a <![CDATA[68.46±8.60 ** ]]> 1b <![CDATA[71.93±7.60 ** ]]> 1c <![CDATA[71.38±7.88 ** ]]> 1d <![CDATA[69.98±4.59 ** ]]> 1e <![CDATA[81.37±10.05 **** ]]> 1f <![CDATA[77.35±2.73 *** ]]> 1g 64.60±4.74 1h <![CDATA[67.68±8.22 * ]]> EDA <![CDATA[68.14±12.93 * ]]> OGD / R 46.63±9.27

[0266] Table 6 Effects of Compounds 1i-1j (20 μM) on the Viability of HT22 Hippocampal Neurons Injured by OGD / R (Oxygen Glucose Deprivation Reperfusion) in Vitro

[0267] Compound Cell viability (% relative to the control group) 1i 44.43±4.46 1j <![CDATA[68.45±4.68 * ]]> EDA <![CDATA[72.21±5.26 ** ]]> OGD / R 49.21±6.98

[0268] The experimental results (Table 5 and Table 6) show that compounds 1a / 1b / 1c / 1d / 1e / 1f / 1h / 1j have a significant protective effect on the OGD / R-induced in vitro cerebral ischemia model at a concentration of 20 μM, and the effect is close to or better than that of the control drug edaravone (EDA); the above compounds can be used to prepare drugs for treating cerebral ischemia-related diseases.

[0269] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that these are only examples, and various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the protection scope of the present invention is limited by the appended claims.

Claims

1. A polyphenol compound as represented by formula X, its stereoisomers or pharmaceutically acceptable salts thereof: in, R1 is C 1-6 Alkyl, C 6-14 Aryl or 5- to 6-membered heteroaryl; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R2 and R3 are independently a hydroxy protecting group or hydrogen; The hydroxyl protecting group is C 1-6 Acyl.

2. The polyphenol compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The polyphenol compound represented by formula X is a compound represented by formula I: Where R1 is C 1-6 Alkyl or 5- to 6-membered heteroaryl; the heteroatoms of the 5- to 6-membered heteroaryl are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3.

3. The polyphenol compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: It meets one or more of the following conditions: (1) C 1-6 Alkyl is methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl or isobutyl, preferably methyl, ethyl, isopropyl, n-propyl or isobutyl, for example ethyl; (2) The 5- to 6-membered heteroaryl group has 1 or 2 heteroatoms, such as pyridyl, preferably pyridin-3-yl or pyridin-4-yl; (3) C 6-14 Aryl is phenyl or naphthyl, for example phenyl; and (4) C 1-6 The acyl group is formyl, acetyl, isopropionyl, n-propionyl, n-butyryl, tert-butyryl or isobutyryl, preferably acetyl.

4. The polyphenol compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: The compound represented by formula X is a compound represented by formula X-1: Wherein, R1, R2 and R3 are independently as described in claim 1 or 2; Preferably, the compound represented by formula I is a compound represented by formula I': Wherein, R1, R2 and R3 are independently as described in claim 1 or 2.

5. The polyphenol compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: It is any of the following compounds:

6. A polyphenol compound, wherein the compound is a compound as shown in formula XI: in, R1' is C 1-6 Alkyl, C 2-6 Alkenyl, C 6-14 Aryl or 5- to 6-membered heteroaryl; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R' is a hydroxyl protecting group; R2' and R3' are hydroxyl protecting groups or hydrogen.

7. The polyphenol compound according to claim 6, characterized in that The compound represented by formula XI is the compound represented by formula II; Where R1' is C 2-6 Alkenyl or 5- to 6-membered heteroaryl; the heteroatoms of the 5- to 6-membered heteroaryl are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R' is a hydroxyl protecting group.

8. The polyphenol compound according to claim 6 or 7, characterized in that: It meets one or more of the following conditions: (1) R' is benzyl; (2) In R2' and R3', the hydroxyl protecting group is C 1-6 Acyl, preferably formyl, acetyl, isopropanoyl, n-propanoyl, n-butyryl, tert-butyryl or isobutyryl, for example acetyl; (3) C 2-6 Alkenyl is vinyl, prop-1-en-2-yl, (E)-prop-1-en-1-yl or 2-methylprop-1-en-1-yl, for example vinyl; (4) C 6-14 Aryl is phenyl or naphthyl, for example phenyl; (5) the 5- to 6-membered heteroaryl group has 1 or 2 heteroatoms, such as pyridyl, preferably pyridin-3-yl or pyridin-4-yl; and (6) C 1-6 The alkyl group is methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl or isobutyl, preferably methyl, ethyl, isopropyl, n-propyl or isobutyl, for example methyl.

9. The polyphenol compound according to claim 6, characterized in that The compound of formula XI is a compound as shown in formula III: R" is a hydroxyl protecting group, such as acetyl; R1' and R' are as described in claim 6.

10. The polyphenol compound according to claim 6, characterized in that It is any of the following compounds:

11. A method for preparing a compound as shown in formula X as claimed in claim 1 or 2, characterized in that: It is any of the following options: Option 1: The method comprises the following steps: under the catalysis of catalyst 1 and in the presence of hydrogen, the compound represented by formula XI is subjected to a reaction of removing the protecting group R' in an organic solvent to obtain a compound represented by formula X; Where R1' is C 1-6 Alkyl, C 6-14 Aryl or 5- to 6-membered heteroaryl; The R', R2', R3', C 1-6 Alkyl, C 6-14 Aryl and 5- to 6-membered heteroaryl are independently as described in any one of claims 6-10; R1, R2 and R3 are independently as described in claim 1 or 2; Scheme 2: The method comprises the following steps: under the catalysis of catalyst 1, in the presence of hydrogen, the compound represented by formula XI is subjected to a reaction of removing the protecting group R' and reducing the olefin group in an organic solvent to obtain a compound represented by formula X, Where R1' is C 2-6 alkenyl; The R', R2', R3', the C 2-6 The alkenyl group is independently as described in any one of claims 6 to 10; R1, R2 and R3 are independently as described in claim 1 or 2; Preferably, the catalyst 1 is 20% palladium hydroxide on carbon; And / or, the organic solvent is methanol.

12. A method for preparing a compound of formula XI as claimed in claim 6 or 7, characterized in that: It is any of the following methods: Method 1: subjecting the compound of formula III as claimed in claim 9 to a reaction of removing the hydroxyl protecting group R" to obtain the compound of formula II as claimed in claim 7; Method 2: In a mixed solvent 2, under the protection of an inert gas, in the presence of a base 2, and under the catalysis of a catalyst 2, a compound represented by formula IV and a R1' donor are subjected to a Suzuki coupling reaction to obtain a compound represented by formula XI as claimed in claim 6; Wherein, R', R2' and R3' are as described in claim 6 or 7; Method 3: In an organic solvent, in the presence of a base 3, in the presence of an acylating agent, and under the catalysis of a catalyst 3, an acylation reaction is performed on the compound of formula II as claimed in claim 7 to obtain a compound of formula XI as claimed in claim 6; In method 1, the R" is preferably acetyl; In method 2, R1' is defined as in claim 6 or 7; the inert gas is nitrogen or argon; the Suzuki coupling reaction is carried out under heating reflux; Preferably, the R1' donor is methylboric acid, potassium ethylene trifluoroborate, isopropenylboronic acid pinacol ester, trans-1-propylene-1-boric acid, isobutenylboronic acid pinacol ester, phenylboric acid, 4-pyridineboric acid or 3-pyridineboric acid; and / or, the catalyst 2 is tetrakistriphenylphosphine palladium; and / or, the mixed solvent 2 is selected from two or three of dioxane, water and acetonitrile, preferably dioxane and water, the volume ratio is 4:1, or dioxane, acetonitrile and water, the volume ratio is 3:3:1; and / or, the base 2 is potassium carbonate; In method three, R2' and R3' are hydrogen atoms; and / or, the organic solvent is preferably dichloromethane; and / or, the base 3 is preferably triethylamine; and / or, the acylating agent is preferably acetic anhydride; and / or, the catalyst 3 is 4-dimethylaminopyridine; More preferably, in method 1, the compound III is prepared by the following method: in the mixed solvent 2, under the protection of the inert gas, in the presence of the base 2, and under the catalysis of the catalyst 2, a compound as shown in formula V and a R1' donor are subjected to a Suzuki coupling reaction; Wherein, R' and R" are as defined in claim 6 or 7; R1' is defined as in claim 6 or 7; the inert gas is nitrogen or argon, and the Suzuki coupling reaction is carried out under heating reflux; More preferably, the compound as shown in formula V is and / or, the R1' donor is potassium ethylene trifluoroborate; and / or, the catalyst 2 is palladium tetrakistriphenylphosphine; and / or, the mixed solvent 2 is dioxane and water, preferably in a volume ratio of 4:1; and / or, the base 2 is potassium carbonate.

13. Use of a polyphenol compound represented by formula X, its stereoisomers, and pharmaceutically acceptable salts thereof in the preparation of a drug for treating ischemic stroke; Preferably, the manifestations of ischemic stroke are one or more of the following: (1) behavioral abnormalities; (2) cerebral infarction; (3) HT22 hippocampal neuron cell damage; in, R1 is C 1-6 Alkyl, C 6-14 Aryl, 5- to 6-membered heteroaryl or H; the heteroatom of the 5- to 6-membered heteroaryl is selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R2 and R3 are independently a hydroxyl protecting group or hydrogen; the hydroxyl protecting group is C 1-6 acyl group; Preferably, the polyphenol compound represented by formula X is a polyphenol compound represented by formula X as claimed in any one of claims 1 to 5.

14. The use according to claim 13, characterized in that The drug further comprises a pharmaceutically acceptable carrier; Alternatively, when R1 is H, R2 and / or R3 are the hydroxyl protecting groups; for example, the polyphenol compound represented by formula X is 15. The use according to claim 13, characterized in that The HT22 hippocampal neuron cell damage is manifested as: one or more of increased inflammatory factors, increased apoptotic factors and cell inactivation; the inflammatory factors are preferably iNOS and / or COX-2; the apoptotic factors are preferably one or more of p53, cleaved-Caspase3 and Bax; the cell inactivation is preferably reduced VCAM-1 protein expression and / or reduced cell viability detected by CCK-8 method.

16. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to claim 13 or 14, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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