Ginkgolide b derivatives and salts thereof, and methods of making and using the same

Abstract

The present application belongs to the technical field of medicine, and relates to derivatives of carboxylic acid groups introduced by 10-position hydroxyl in the structure of ginkgolide B and ester derivatives of the carboxylic acid groups, and pharmaceutically acceptable organic or inorganic salts, as shown in formula 1 and formula 2. The present application takes ginkgolide B as a matrix, and is chemically modified to improve the solubility, increase the bioavailability, and enhance the curative effect of ginkgolide B. The prepared compound and carboxylic acid salt thereof have significant platelet activating factor antagonism, anticoagulation effect, and acute cerebral ischemia effect, and can be used for preparing medicines for preventing and treating ischemic stroke, thrombosis, angina pectoris, cardiopulmonary infarction, and diseases related to platelet activating factor, such as inflammation, asthma, and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and relates to ginkgolide B carboxylic acid derivatives and esters thereof, and corresponding salts of the ginkgolide B carboxylic acid derivatives, in particular to the ginkgolide B carboxylic acid derivatives and esters thereof, and a preparation method and use thereof. The present application takes ginkgolide B as a parent body, and modifies the chemical structure to prepare ginkgolide B derivatives, so as to change the solubility, improve the bioavailability, and enhance the curative effect. BACKGROUND

[0002] It is recorded that Chinese folk used ginkgo leaves to treat asthma and bronchitis around 1000 AD. In modern times, with the standardization of drug extraction technology and the in-depth research on the activity of pharmacological action, ginkgo biloba extract (GBE) has been widely used in the treatment of respiratory system and cardiovascular system and other diseases in countries all over the world, especially in European countries such as Germany and France.

[0003] The prior art discloses that ginkgolide B (GB) is a diterpenoid compound extracted from ginkgo leaves; biological activity evaluation shows that ginkgolide B is the most active natural product of platelet activating factor (PAF) antagonist ever found. Platelet activating factor can promote platelet and neutrophil aggregation, participate in various inflammatory reaction processes, thereby increasing vascular permeability, promoting thrombosis, inducing smooth muscle contraction, and playing an important role in the occurrence and development of a series of related diseases such as inflammation, asthma, cardiovascular and cerebrovascular microcirculation disorders, and gastrointestinal mucosal damage. Meanwhile, it is found that ginkgolide B also has strong anti-inflammatory effect. In the inflammatory reaction, the membrane phospholipid of neutrophils is hydrolyzed into arachidonic acid (AA) by the action of LPA of activated phospholipase A2, and AA is further metabolized into leukotrienes (LTs) and hydroxyeicosatetraenoic acid (HETEs) and other products under the action of 5-lipoxygenase (5-LO), wherein, certain products are important inflammatory mediators, and the activation of phospholipase A2 needs the participation of intracellular calcium, ginkgolide B has the effect of affecting the arachidonic acid metabolism enzyme of rat neutrophils and intracellular free calcium, and its anti-inflammatory effect may be related to the inhibition of the release of neutrophil lysosomal enzyme, the production of superoxide anion and the increase of intracellular calcium level.

[0004] At present, the ginkgolide drugs used in clinical mainly include two categories of ginkgolide mixed extract containing ginkgolide B component and ginkgolide B as the main component, which are mainly used for the treatment of thrombosis, acute pancreatitis and cardiovascular diseases, and the treatment of metastatic cancer, and the protection of damaged neurons. However, practice shows that due to the structural characteristics of the six-ring cage diterpenoid compound of ginkgolide B, the structure is relatively rigid, the water solubility is poor, and the bioavailability is also poor, so that the full play of its drug efficacy is limited, which affects the clinical application effect.

[0005] Based on the status quo of the prior art, the inventors of the present application propose ginkgolide B carboxylic acid derivatives and esters thereof, and a preparation method and use thereof. The present application uses ginkgolide B as a matrix, and modifies the chemical structure to prepare ginkgolide B derivatives to change the water solubility, improve the bioavailability, and enhance the efficacy. SUMMARY

[0006] The present application aims to provide a new type of ginkgolide B carboxylic acid derivatives and ester compounds thereof. The ginkgolide B carboxylic acid derivatives can be prepared into corresponding organic or inorganic base salts. The present application introduces various carboxyl-containing groups and ester groups on the 10-hydroxyl group of ginkgolide B through structural modification, to obtain a new type of ginkgolide B carboxylic acid derivatives and esters, which can improve the solubility and overcome the poor bioavailability of ginkgolide B and other adverse factors.

[0007] The present application provides a compound as shown in formula 1,

[0008]

[0009] wherein,

[0010] L is a heteroatom, a substituted or unsubstituted C 1-10 alkylene group, a substituted or unsubstituted heteroatom-containing C 1-10 alkylene group, or nothing, when L is a heteroatom or when L is a substituted or unsubstituted heteroatom-containing C 1-10 alkylene group, the heteroatom is selected from one or more of oxygen, nitrogen and sulfur; when the heteroatom is more than one, the heteroatoms are the same or different;

[0011] R is hydrogen, or a substituted or unsubstituted C 1-8 alkyl group;

[0012] wherein the substituted C 1-10 alkylene group, the substituted heteroatom-containing C 1-10 alkylene group, and the substituted C 1-8 alkyl group are independently substituted with one or more of halogen, hydroxyl, C 1-10 alkoxy, phenyl and C 1-10 alkyl, and when there are more than one substituent, the substituents are the same or different;

[0013] the compound shown in formula I is not:

[0014] In a preferred embodiment of the present application, when the substituted C 1-10 alkylene group, the substituted heteroatom-containing C 1-10 alkylene group, and the substituted C 1-8When the substituents in the hydrocarbyl group are halogen, the halogen is preferably fluorine, chlorine, bromine or iodine;

[0015] In a preferred embodiment of the present application, when the substituted C 1-10 alkylene, the substituted heteroatom-containing C 1-10 heteroalkylene, the substituted C 1-8 alkyl, the substituted C 1-10 alkoxy, the substituted C 1-10 alkoxy is preferably C 1-4 alkoxy, more preferably methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy or t-butoxy.

[0016] In a preferred embodiment of the present application, when the substituted C 1-10 alkylene, the substituted heteroatom-containing C 1-10 heteroalkylene, the substituted C 1-8 alkyl, the substituted C 1-10 alkyl, the substituted C 1-10 alkyl is preferably C 1-4 alkyl, more preferably methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl.

[0017] In the present application, the term alkylene refers to a chain alkylene group, wherein the number of carbon-carbon double bonds is one or more, and the carbon-carbon double bonds can be located at any position of the chain alkylene group.

[0018] In the present application, the term alkenylene refers to a chain alkenylene group, wherein the number of carbon-carbon double bonds is one or more, and the carbon-carbon double bonds can be located at any position of the chain alkenylene group.

[0019] In the present application, the term alkynylene refers to a chain alkynylene group, wherein the number of carbon-carbon triple bonds is one or more, and the carbon-carbon triple bonds can be located at any position of the chain alkynylene group.

[0020] In the present application, the term cycloalkenylene refers to a cyclic alkenylene group, wherein the number of carbon-carbon double bonds is one or more, and the carbon-carbon double bonds can be located at any position of the cyclic alkenylene group.

[0021] In the present application, the term cycloalkynylene refers to a cyclic alkynylene group, wherein the number of carbon-carbon triple bonds is one or more, and the carbon-carbon triple bonds can be located at any position of the cyclic alkynylene group.

[0022] In the present application, the term hydrocarbyl refers to a group containing carbon and hydrogen atoms, which is left after losing any one hydrogen atom from the corresponding hydrocarbon. The term hydrocarbyl is preferably alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl or aryl.

[0023] In the present application, the term alkenyl refers to a chain alkenyl group, wherein the number of carbon-carbon double bonds is one or more, and the carbon-carbon double bonds can be located at any position of the chain alkenyl group.

[0024] In the present application, the term alkynyl refers to a chain alkynyl group, wherein the number of carbon-carbon triple bonds is one or more, and the carbon-carbon triple bonds can be located at any position of the chain alkynyl group.

[0025] In the present application, the term cycloalkenyl refers to a cyclic alkenyl group, wherein the number of carbon-carbon double bonds is one or more, and the carbon-carbon double bonds can be located at any position of the cyclic alkenyl group.

[0026] In the present application, the term cycloalkynyl refers to a cyclic alkynyl group, wherein the number of carbon-carbon triple bonds is one or more, and the carbon-carbon triple bonds can be located at any position of the cyclic alkynyl group.

[0027] In the present application, the term heteroalkylene refers to the insertion of one or more heteroatoms at any position of the alkylene group in the present application. The definition of the alkylene group is the same as described above.

[0028] In the present application, the term chain includes straight chain and branched chain.

[0029] In the present application, the term cycloalkyl preferably refers to C3-C6cycloalkyl, more preferably C3-C4cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. 10 cycloalkyl, more preferably C3-C4cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0030] In the present application, the term aryl preferably refers to C6-C10aryl, more preferably C6-C8aryl. Examples of aryl include, but are not limited to, phenyl, naphthyl or tetrahydronaphthyl. 10 cycloalkyl, more preferably C3-C4cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0031] In the present application, the term heteroalkylene refers to the insertion of one or more heteroatoms at any position of the alkylene group in the present application. The definition of the alkylene group is the same as described above. 10 heteroalkylene, further preferably C1-C8heteroalkylene, more preferably C1-C6heteroalkylene, wherein the number of heteroatoms is 1, 2, 3 or 4, and the heteroatoms are selected from O, N and S.

[0032] In the present application, the term alkyl includes branched and straight chain saturated aliphatic hydrocarbon groups, preferably 1-10 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, 4,4-dimethylpentyl, 2,2,4-trimethylpentyl, and various isomers thereof. In the present application, alkyl is preferably C1-C4alkyl, more preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or t-butyl.

[0033] In a preferred embodiment of the invention, the "substituted or unsubstituted C" 1-10 C in "hydro-ion group" 1-10 The preferred hydrocarbon group is C. 1-10 Alkylene, C 2-10 imidene group, C 2-10 Ethyne group, C 3-10 Cycloalkylene, C 3-10 Cycloalkylene, C 4-10 Cycloalkynyl, phenylene, or naphthylene.

[0034] In a preferred embodiment of the invention, the "substituted or unsubstituted heteroatom-containing C" 1-10 C in "heterohydrocarbon group" 1-10 The heteroalkyl group is preferably C 1-10 Heteroalkylene, C 2-10 Heteroeneyl, C 2-10 Hetero-ynyl group, C 2-10 Heterocyclic alkyl, C 2-10 Heterocyclic alkenyl, C 2-10 Heterocyclic alkyne or C 1-10 Hybrid aryl.

[0035] In a preferred embodiment of the invention, the "substituted or unsubstituted C" 1-8 The C in "hydrocarbon group" 1-8 The preferred hydrocarbon group is C. 1-8 Alkyl, C 2-8 alkenyl, C2-8 ynyl, C 3-8 cycloalkyl, C 3-8 Cycloalkenyl, C 4-8 Cycloalkynyl or phenyl.

[0036] In a preferred embodiment of the invention, the L is preferably a heteroatom, a substituted or unsubstituted C. 1-10 Alkylene, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 alkyne-based, substituted or unsubstituted C 3-10 Cycloalkylene, substituted or unsubstituted C 3-10 Cycloalkenyl, substituted or unsubstituted C 4-10 Cycloalkynyl, substituted or unsubstituted phenylene, substituted or unsubstituted naphthylene, substituted or unsubstituted C 1-10 Heteroalkylene, substituted or unsubstituted C 2-10 Heteroene-based, substituted or unsubstituted C 2-10 Hetero-ynyl, substituted or unsubstituted C 2-10 Heterocyclic alkyl, substituted or unsubstituted C 2-10 Heterocyclic alkenyl, substituted or unsubstituted C 2-10 Heterocyclic alkyne group, substituted or unsubstituted C1-10 heteroarylene, or nothing.

[0037] In a preferred embodiment of the present application, R is preferably hydrogen, methyl, benzyl, substituted or unsubstituted C 3-8 alkyl, substituted or unsubstituted C 2-8 alkenyl, substituted or unsubstituted C 2-8 alkynyl, substituted or unsubstituted C 3-8 cycloalkyl, substituted or unsubstituted C 3-8 cycloalkenyl, substituted or unsubstituted C 4-8 cycloalkynyl, or substituted or unsubstituted phenyl.

[0038] In a preferred embodiment of the present application, when L is a heteroatom, the heteroatom is preferably oxygen or sulfur.

[0039] In a preferred embodiment of the present application, when L is substituted or unsubstituted C 1-10 alkylene, the C 1-10 alkylene in "substituted or unsubstituted C 1-10 alkylene" is preferably C 1-5 alkylene, further preferably methyl, ethyl, n-propylene, i-propylene, n-butylene, i-butylene, t-butylene, or n-pentylene.

[0040] In a preferred embodiment of the present application, when L is substituted or unsubstituted C 2-10 alkenylene, the C 2-10 alkenylene in "substituted or unsubstituted C 2-10 alkenylene" is preferably C 2-5 alkenylene, further preferably ethenylene, propenylene, butenylene or pentenylene, more further preferably ethenylene, most preferably -CH=CH-.

[0041] In a preferred embodiment of the present application, when L is substituted or unsubstituted C 2-10 alkynylene, the C 2-10 alkynylene in "substituted or unsubstituted C 2-10 alkynylene" is preferably C 2-5 alkynylene, further preferably ethynylene, propynylene, butynylene or pentynylene.

[0042] In a preferred embodiment of the present application, when L is substituted or unsubstituted C 3-10 cycloalkylene, the C 3-10 cycloalkylene in "substituted or unsubstituted C 3-10 cycloalkylene" is preferably C 3-8Cycloalkylene, further preferred cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene or cyclooctylene.

[0043] In a preferred embodiment of the present application, when L is a substituted or unsubstituted C 3-10 cycloalkylene, further preferred cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene or cyclooctylene. 3-10 cycloalkylene, further preferred cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene or cyclooctylene. 3-10 cycloalkylene, further preferred cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene or cyclooctylene. 3-8 cycloalkylene, further preferred cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene or cyclooctylene.

[0044] In a preferred embodiment of the present application, when L is a substituted or unsubstituted C 1-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 1-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 1-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 1-6 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 1-4 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 1-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C

[0045] In a preferred embodiment of the present application, when L is a substituted or unsubstituted C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-5 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C

[0046] In a preferred embodiment of the present application, when L is a substituted or unsubstituted C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-10 heteroalkylene, further preferred heteroalkylene, wherein the heteroatom is preferably oxygen or sulphur and the number of heteroatoms is preferably 1 or 2, and the C 2-4The heteroynyl group, more preferably heteroethynyl, heteropropynyl, heterobutynyl, or heteropentynyl, wherein the heteroatom is preferably oxygen or sulfur, and the number of heteroatoms is preferably 1 or 2. 2-10 The most preferred heteroynyl group is -C≡CO-CH2-, -C≡CO-, -CH2OC≡COCH2-, or -C≡CS-CH2-.

[0047] In a preferred embodiment of the invention, when L is substituted or unsubstituted C 2-10 When it is a heterocycloalkylene, the "substituted or unsubstituted C" refers to... 2-10 C in "heterocyclic alkyl" 2-10 Heterocyclic alkyl groups are preferably C 2-5 The heterocyclic alkyl group is more preferably heterocyclic ethyl, heterocyclic propyl, heterocyclic butyl, or heterocyclic pentyl, wherein the heteroatom is preferably oxygen or sulfur, and the number of heteroatoms is preferably 1 or 2.

[0048] In a preferred embodiment of the invention, when L is substituted or unsubstituted C 2-10 When heterocyclic alkenyl, the "substituted or unsubstituted C" refers to... 2-10 C in "heterocyclic alkenyl" 2-10 Heterocyclic alkenyl groups are preferably C 2-5 The heterocyclic alkenyl group is more preferably a heterocyclic vinyl group, a heterocyclic propenyl group, a heterocyclic butenyl group, or a heterocyclic pentenyl group, wherein the heteroatom is preferably oxygen or sulfur, and the number of heteroatoms is preferably 1 or 2.

[0049] In a preferred embodiment of the invention, when L is substituted or unsubstituted C 1-10 In the case of heteroarylene, the "substituted or unsubstituted C" refers to... 1-10 C in "heteroarylene" 1-10 Heteroaryl is preferably C 1-9 The heteroaryl group, wherein the heteroatom is preferably oxygen or nitrogen, and the number of heteroatoms is preferably 1, 2, 3 or 4.

[0050] In a preferred embodiment of the invention, when R is substituted or unsubstituted C 1-8 When alkyl, the "substituted or unsubstituted C" refers to... 3-8 C in "alkyl" 3-8 The alkyl group is preferably n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.

[0051] In a preferred embodiment of the invention, when R is substituted or unsubstituted C 3-8 When alkyl, the "substituted or unsubstituted C" refers to... 3-8 C in "alkyl" 3-8 The alkyl group is preferably n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.

[0052] In a preferred embodiment of the present application, when R is substituted or unsubstituted C 2-8 alkenyl, the C 2-8 alkenyl in "substituted or unsubstituted C 2-8 alkenyl" is preferably C 2-5 alkenyl, further preferably ethenyl, propenyl, butenyl or pentenyl.

[0053] In a preferred embodiment of the present application, when R is substituted or unsubstituted C 2-8 alkynyl, the C 2-8 alkynyl in "substituted or unsubstituted C 2-8 alkynyl" is preferably C 2-5 alkynyl, further preferably ethynyl, propynyl, butynyl or pentynyl.

[0054] In a preferred embodiment of the present application, when R is substituted or unsubstituted C 3-8 cycloalkyl, the C 3-8 cycloalkyl in "substituted or unsubstituted C 3-8 cycloalkyl" is preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl.

[0055] In a preferred embodiment of the present application, when R is substituted or unsubstituted C 3-8 cycloalkenyl, the C 3-8 cycloalkenyl in "substituted or unsubstituted C 3-8 cycloalkenyl" is preferably cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl or cyclooctenyl.

[0056] In a preferred embodiment of the present application, L is preferably substituted or unsubstituted C 2-10 alkenylene, substituted or unsubstituted C 1-10 heteroalkylene, or is absent.

[0057] In a preferred embodiment of the present application, R is preferably hydrogen, methyl, benzyl, or substituted or unsubstituted C 3-8 alkyl.

[0058] In a preferred embodiment of the present application, L is preferably -CH=CH-, -CH2OCH2-, -CH2OCH2CH2OCH2-, or L is absent.

[0059] In a preferred embodiment of the present application, R is preferably hydrogen, methyl, or tert-butyl.

[0060] In a preferred embodiment of the present application, the compound of Formula 1 or a pharmaceutically acceptable salt thereof is selected from any one of the following compounds:

[0061]

[0062] The present application also provides a compound of Formula 2:

[0063]

[0064] wherein L is as defined above;

[0065] Cations, including cations formed from various inorganic and organic bases; the inorganic cations can be preferably selected from sodium, potassium, calcium, magnesium, zinc, or ammonium, but not limited to these salts; the organic cations can be preferably selected from substituted or unsubstituted C 1-8 Primary amines having substituted or unsubstituted C 1-8 Secondary amines having substituted or unsubstituted C 1-8 Tertiary amines having substituted or unsubstituted C

[0066] In the present application, the primary amines having substituted or unsubstituted C 1-8 hydrocarbyl groups substituted for the hydrogen atoms on ammonia; wherein the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C 1-8 hydrocarbyl groups are as defined above.

[0067] In the present application, the secondary amines having substituted or unsubstituted C 1-8 hydrocarbyl groups substituted for the hydrogen atoms on ammonia; wherein the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C 1-8 hydrocarbyl groups can be cyclized in any manner (without violating the common sense of the art) to form rings such as pyrrolidine, piperidine, morpholine, or thiomorpholine; wherein the substituted or unsubstituted C 1-8 hydrocarbyl groups are as defined above.

[0068] In the present application, the tertiary amines having substituted or unsubstituted C 1-8 hydrocarbyl groups substituted for the hydrogen atoms on ammonia; wherein the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C 1-8 hydrocarbyl groups can be the same or different, and the substituted or unsubstituted C1-8 The hydrocarbon group can be cyclized in any way (without departing from the common knowledge in the art) to form a ring such as a pyrrolidine, piperidine, morpholine or thiomorpholine substituted with a nitrogen atom; wherein the substituted or unsubstituted C 1-8 The hydrocarbon group is as described above.

[0069] In the present application, the natural or unnatural amino acid refers to an amine containing a carboxyl group in the hydrocarbon chain, and is preferably L-arginine, L-histidine, L-lysine, D-arginine, D-histidine, D-lysine.

[0070] In a preferred embodiment of the present application, the inorganic base in the cation formed by various inorganic bases and organic bases is preferably sodium carbonate, sodium bicarbonate, sodium hydroxide, potassium carbonate, calcium hydroxide, magnesium carbonate, magnesium hydroxide, ammonium hydroxide or zinc hydroxide.

[0071] In a preferred embodiment of the present application, the organic base in the cation formed by various inorganic bases and organic bases can be an amine compound or a natural or unnatural amino acid compound. The amine compound is particularly preferably methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, propylamine, butylamine, pentylamine, hexylamine, N-methyl-D-glucosamine, pyrrolidine, morpholine and thiomorpholine. The amino acid compound is preferably L-lysine, L-histidine, L-arginine, D-lysine, D-histidine and D-arginine.

[0072] In a preferred embodiment of the present application, the cation in the compound of formula 2 is preferably

[0073]

[0074]

[0075] In a preferred embodiment of the present application, the compound of formula 2 is selected from any one of the following compounds:

[0076]

[0077]

[0078] The present application also provides a preparation method of the compound of formula 1 or a pharmaceutically acceptable salt thereof, which comprises the following steps:

[0079] The ether reaction of ginkgolide B and the compound of formula 3 in the presence of a base and a catalyst as shown below can obtain the compound of formula 1,

[0080]

[0081] wherein R and L are as defined above, and X is halogen.

[0082] In the ether-forming reaction, X is preferably fluorine, chlorine, bromine or iodine, and is more preferably chlorine, bromine or iodine.

[0083] In the ether-forming reaction, the organic solvent can be any conventional organic solvent for such reactions in the art, and the present application particularly preferably comprises one or more of a halogenated hydrocarbon solvent, an ether solvent, a ketone solvent, a nitrile solvent and an amide solvent. The halogenated hydrocarbon solvent is preferably dichloromethane and / or chloroform. The ether solvent is preferably tetrahydrofuran and / or dioxane, and is more preferably tetrahydrofuran. The ketone solvent is preferably acetone and / or butanone. The nitrile solvent is preferably acetonitrile. The amide solvent is preferably N,N-dimethylformamide.

[0084] In the ether-forming reaction, the base can be any conventional base for such reactions in the art, such as an organic base and / or an inorganic base. The organic base is preferably an amine organic base, and is more preferably one or more of triethylamine, pyrrolidine and piperazine. The inorganic base is preferably a carbonate and / or a bicarbonate. The carbonate is preferably one or more of sodium carbonate, potassium carbonate and cesium carbonate. The bicarbonate is preferably sodium bicarbonate.

[0085] In the ether-forming reaction, the catalyst can be any conventional catalyst for such reactions in the art, and the present application particularly preferably comprises one or more of an iodide, and is more preferably one or more of potassium iodide, sodium iodide and cuprous iodide, and is most preferably potassium iodide.

[0086] In the ether-forming reaction, the molar concentration of ginkgolide B in the organic solvent can be any conventional molar concentration for such reactions in the art, and the present application particularly preferably comprises 0.001 to 1 mol / L, and is more preferably 0.001 to 0.5 mol / L, and is even more preferably 0.001 to 0.1 mol / L (e.g. 0.025 mol / L, 0.024 mol / L, 0.023 mol / L).

[0087] In the ether-forming reaction, the molar ratio of ginkgolide B to the compound of formula 3 can be any conventional molar ratio for such reactions in the art, and the present application particularly preferably comprises 1:1 to 1:5, and is more preferably 1:1 to 1:3 (e.g. 1:2).

[0088] In the ether-forming reaction, the molar ratio of ginkgolide B to the base can be any conventional molar ratio for such reactions in the art, and the present application particularly preferably comprises 1:1 to 1:10, and is more preferably 1:3 to 1:6 (e.g. 1:4.6, 1:4.4).

[0089] The molar ratio of ginkgolide B to catalyst in the ether-forming reaction can be the conventional molar ratio in the art, and the present application particularly preferably is 1:1 to 1:5, and further preferably is 1:1 to 1:3 (e.g., 1:2).

[0090] The reaction progress in the ether-forming reaction can be monitored by conventional monitoring methods in the art (e.g., TLC, HPLC or NMR), and generally the reaction endpoint is taken as the time when ginkgolide B is substantially consumed. The reaction time in the present application particularly preferably is 1 to 5 hours, and further preferably is 1 to 3 hours (e.g., 2 hours).

[0091] The reaction temperature in the ether-forming reaction can be the conventional reaction temperature in the art, and the present application particularly preferably is the temperature at which the organic solvent used is refluxed at normal temperature and pressure.

[0092] In a preferred embodiment of the present application, the ether-forming reaction comprises the following steps: after mixing ginkgolide B and the organic solvent, the compound of formula 3, the catalyst and the base are sequentially added to react.

[0093] In a preferred embodiment of the present application, the ether-forming reaction further comprises a post-treatment step after the reaction is completed. The post-treatment step can be the conventional post-treatment step in the art, and preferably is filtration, concentration and purification. The purification method can be the conventional purification method in the art (e.g., column chromatography).

[0094] The present application also provides a preparation method of the compound of formula 2, which comprises the following steps:

[0095] In an organic solvent, the compound of formula 1 is subjected to hydrolysis under the action of an acid or a base to obtain a compound of formula 1 in which R is hydrogen; and the compound is subjected to salt formation with various bases to obtain various salts of formula 2.

[0096]

[0097] wherein L and the cation are as defined above;

[0098] R is a substituted or unsubstituted C 1-8 hydrocarbon group, and the "substituted or unsubstituted C 1-8 hydrocarbon group" is as defined above.

[0099] In the hydrolysis reaction, when the compound of Formula 1 is subjected to the hydrolysis reaction in the presence of a base, the organic solvent can be a conventional organic solvent for such a reaction in the art, and the present application particularly preferably is an alcohol solvent, and further preferably is anhydrous alcohol solvent. The alcohol solvent preferably is one or more of methanol, ethanol, isopropanol and tert-butanol, and further preferably is methanol. The organic solvent preferably is anhydrous methanol.

[0100] In the hydrolysis reaction, when the compound of Formula 1 is subjected to the hydrolysis reaction in the presence of a base, the base can be a conventional base for such a reaction in the art, such as inorganic base and / or organic base. The inorganic base preferably is one or more of hydroxide, carbonate and bicarbonate. The hydroxide preferably is one or more of lithium hydroxide, sodium hydroxide and potassium hydroxide. The carbonate preferably is one or more of sodium carbonate, potassium carbonate and cesium carbonate. The bicarbonate preferably is one or more of sodium bicarbonate, potassium bicarbonate and lithium bicarbonate. The organic base preferably is amine organic base and / or imine organic base, and further preferably is one or more of triethylamine, pyridine, pyrrolidine and piperazine.

[0101] In the hydrolysis reaction, when the compound of Formula 1 is subjected to the hydrolysis reaction in the presence of a base, the molar ratio of the compound of Formula 1 to the base can be a conventional molar ratio for such a reaction in the art, and the present application particularly preferably is 1:1 to 1:10, and further preferably is 1:2 to 1:4 (e.g. 1:2.08, 1:2.78, 1:3.53).

[0102] In the hydrolysis reaction, when the compound of Formula 1 is subjected to the hydrolysis reaction in the presence of a base, the reaction temperature of the hydrolysis reaction can be a conventional reaction temperature for such a reaction in the art, and the present application particularly preferably is a temperature at which the used organic solvent is refluxed at normal temperature and normal pressure.

[0103] In the hydrolysis reaction, when the compound of Formula 1 is subjected to the hydrolysis reaction in the presence of a base, the reaction progress can be monitored by a conventional monitoring method in the art (e.g. TLC, HPLC or NMR), and generally the reaction endpoint is monitored when the compound of Formula 1 substantially disappears. The reaction time of the present application particularly preferably is 1 to 5 hours, and further preferably is 3 to 4 hours (e.g. 3 hours).

[0104] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of a base, the molar concentration of the compound of formula 5 in the organic solvent can be the conventional molar concentration of this type of reaction in the art, and the present application particularly preferably is 0.001-1 mol / L, further preferably is 0.001-0.5 mol / L, and still further preferably is 0.0034-0.16 mol / L (e.g. 0.034 mol / L, 0.06 mol / L, 0.16 mol / L).

[0105] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of an acid, the organic solvent can be the conventional organic solvent of this type of reaction in the art, and the present application particularly preferably is a halogenated hydrocarbon solvent, further preferably is one or more of dichloromethane, chloroform and carbon tetrachloride, and most preferably is chloroform.

[0106] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of an acid, the acid can be the conventional acid of this type of reaction in the art, such as an inorganic acid and / or an organic acid. The inorganic acid preferably is one or more of hydrochloric acid, phosphoric acid, sulfuric acid and nitric acid. The organic acid preferably is one or more of trifluoroacetic acid, p-toluenesulfonic acid and methanesulfonic acid.

[0107] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of an acid, the molar ratio of the compound of formula 5 to the acid can be the conventional molar ratio of this type of reaction in the art, and the present application particularly preferably is 1:2-1:100, further preferably is 1:50-1:70 (e.g. 1:61.5).

[0108] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of an acid, the molar concentration of the compound of formula 5 in the organic solvent can be the conventional molar concentration of this type of reaction in the art, and the present application particularly preferably is 0.001-1 mol / L, further preferably is 0.001-0.5 mol / L (e.g. 0.219 mol / L).

[0109] In the hydrolysis reaction, when the compound of formula 1 is subjected to the hydrolysis reaction under the action of an acid, the reaction temperature of the hydrolysis reaction can be the conventional reaction temperature of this type of reaction in the art, and the present application particularly preferably is 15-25°C.

[0110] The hydrolysis reaction can be monitored by conventional monitoring methods in the art (e.g. TLC, HPLC or NMR), and the reaction is generally terminated when the compound of formula 5 is substantially consumed. The reaction time is particularly preferably 1-5 hours, and more preferably 1-4 hours (e.g. 2 hours).

[0111] In a preferred embodiment of the present application, the hydrolysis reaction comprises the following steps: mixing the compound of formula 1 with the organic solvent, and adding the acid or base.

[0112] In a preferred embodiment of the present application, the hydrolysis reaction further comprises a post-treatment step after the reaction is terminated. The post-treatment step can be conventional post-treatment steps in the art. When the compound of formula 1 is subjected to the hydrolysis reaction in the presence of an acid, the post-treatment step is preferably concentration. When the compound of formula 1 is subjected to the hydrolysis reaction in the presence of a base, the post-treatment step is preferably pH adjustment of the reaction solution to 3-4 after the reaction is terminated, concentration and filtration. The reagent used for the pH adjustment can be conventional acids in the art, and dilute hydrochloric acid is particularly preferred.

[0113] The present application also provides a method for preparing the compound of formula 2, which comprises the following steps: mixing the compound of formula 1 (R is hydrogen) with a base in an organic solvent to perform a salt formation reaction.

[0114] The organic solvent used in the salt formation reaction can be conventional organic solvents in the art, and alcohol solvents are particularly preferred, and further preferably a mixture of ethanol and methanol.

[0115] The molar concentration of the compound of formula 1 (R is hydrogen) in the organic solvent used in the salt formation reaction can be conventional molar concentrations in the art, and particularly preferably 0.01-0.1 mol / L, and further preferably 0.06-0.07 mol / L (e.g. 0.068 mol / L).

[0116] The inorganic base used in the salt formation reaction is as described above.

[0117] The organic base used in the salt formation reaction is as described above.

[0118] The molar ratio of the compound of formula 1 (R is hydrogen) to the base used in the salt formation reaction can be conventional molar ratios in the art, and particularly preferably 1:1-3.

[0119] The reaction temperature of the salt formation reaction can be the conventional reaction temperature for such reactions in the art, and the present application particularly preferably is 15 to 40°C.

[0120] The progress of the salt formation reaction can be monitored using conventional monitoring methods in the art (e.g., TLC, HPLC, or NMR), and the reaction is generally terminated when the compound of formula 1 (R is a hydrogen atom) is substantially consumed. The reaction time of the present application particularly preferably is 1 to 4 hours, and further preferably is 1 to 2 hours (e.g., 1 hour, 2 hours).

[0121] In a preferred embodiment of the present application, the salt formation reaction comprises the step of mixing the compound of formula 1 (R is a hydrogen atom) with the organic solvent, and adding the organic solvent in which the base is dissolved.

[0122] In a preferred embodiment of the present application, the salt formation reaction further comprises a post-treatment step after the reaction is terminated. The post-treatment step can be a conventional post-treatment step in the art. The present application particularly preferably is recrystallization or direct concentration. The solvent used for recrystallization can be a conventional organic solvent in the art (e.g., diethyl ether).

[0123] The present application also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutically acceptable salt of the compound of formula 2 as described above.

[0124] The present application also provides the use of the compound of formula 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutically acceptable salt of the compound of formula 2 as described above, or the pharmaceutical composition as described above, for the manufacture of a medicament for preventing and treating ischemic stroke, thrombosis, angina pectoris, pulmonary embolism, and inflammation or asthma, and other diseases associated with platelet-activating factor.

[0125] The present application also provides a method for preventing and / or treating ischemic stroke, thrombosis, angina pectoris, pulmonary embolism, and inflammation or asthma, and other diseases associated with platelet-activating factor, which comprises administering to a subject a therapeutically effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutically acceptable salt of the compound of formula 2 as described above, or the pharmaceutical composition as described above.

[0126] The term "pharmaceutically acceptable salt" means a salt of a compound of this application which is found to possess the specific substituent groups discovered in this application, prepared from a relatively non-toxic base. Alkaline earth addition salts of compounds of this application that contain relatively acidic functionalities can be obtained by contacting the neutral form of such compounds in either pure form or in a suitable inert solvent with a sufficient amount of the base in either neat form or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, magnesium, zinc, ammonium, organic amino, or salts of amino acids or similar salts.

[0127] The term "therapeutically effective amount" means a sufficient amount of a drug or pharmaceutical agent to provide the desired effect, which is nontoxic to the subject. Determination of a therapeutically effective amount is within the capability of those skilled in the art, depending on the age and general condition of the subject, as well as the particular active agent. An appropriate therapeutically effective amount in a given case can be determined by one of ordinary skill in the art through routine trials.

[0128] The term "subject" means any animal, preferably a mammal, and most preferably a human, to which the compounds or pharmaceutical compositions according to the embodiments of the present application are administered. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, and the like, with humans being most preferred.

[0129] In the present application, room temperature means 15-25°C. Normal temperature means 25°C. Normal pressure means one atmosphere, 101 KPa. Overnight means 12-18 hours. BRIEF DESCRIPTION OF DRAWINGS

[0130] Figure 1 Example 1-21 Results of in vitro anti-platelet aggregation activity test (n=5, 1 μM)

[0131] Results: Each of the compounds of the examples showed significant inhibitory effect on platelet aggregation. The platelet aggregation rate of ginkolide B at a concentration of 1 μM was 63.42%, and the platelet aggregation rates of each of the compounds of the examples were lower than that of ginkolide B.

[0132] Figure 2 Example 2, 7, 10, 12, 13, 14, 17 and 21 Results of evaluation of in vivo anti-platelet aggregation activity in SD rats (n=10, 50 mg / kg)

[0133] Results: Each of the examples showed significant anti-platelet aggregation activity in SD rats. Except for Example 10, the activities of the compounds of the other examples were higher than that of ginkolide B. Among them, the in vivo activities of Examples 12, 13 and 14 were particularly significant (P<0.001), and can be applied to clinical anticoagulation and treatment of related diseases.

[0134] Figure 3Example 2, 7, 10, 12, 13, 14, 17 and 21 reduced the proportion of the half brain infarction area in SD rats (n=10, 50 mg / kg)

[0135] Results: Compared with the model group, each of the example compounds can significantly reduce the proportion of the half brain infarction area in rats. Except for Example 10, the proportion of the half brain infarction area in the remaining groups of example compounds is less than that of ginkolide B group, and the activities of Examples 12, 13 and 14 are particularly significant (P<0.001), which can be applied to the treatment of clinical cerebral ischemia and cerebral ischemia related diseases.

[0136] Figure 4 . Part of the slice photos of the blank model group, ginkolide B, Example 10, and Example 12 (the original drawing is in color)

[0137] Results: Example 12 is significantly better than ginkolide B and Example 10, and can significantly reduce the proportion of the half brain infarction area in acute cerebral ischemia SD rats, and can be used for the treatment of clinical anticoagulation and anti-cerebral ischemia, and related diseases. DETAILED DESCRIPTION

[0138] Example 1 Preparation of 10-O-(methoxycarbonylmethyl) ginkolide B

[0139]

[0140] Ginkolide B 212 mg (0.5 mmol) was dissolved in 20 mL THF, and chloroacetic acid methyl ester 190 mg (2.0 mmol), KI 166 mg (1.0 mmol), potassium carbonate 310 mg (2.3 mmol) were added in turn. After stirring for 2 hours under heating reflux, the substrate was basically disappeared by plate tracking, and the reaction was ended. After treatment, the potassium carbonate was removed by filtration, the mother liquor was concentrated, and the residue was column chromatographed to obtain a light yellow solid. The product was obtained by filtration and drying, and the yield was 121 mg, 48%. 1H-NMR (DMSO-d6, 400 MHz): 1.01 (s, 9H, t-Bu), 1.08 (d, 3H, 14-Me), 1.74 (dd, 1H, 8-H), 1.87 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.73 (s, 1H, -OCH3), 4.06 (m, 1H, 1-H), 4.42 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.80 (d, 1H, J = 16 Hz, 10-CH2-), 5.15 (d, 1H, 1-OH), 5.29 (s, 1H, 10-H), 5.33 (d, 1H, 6-H), 5.65 (m, 1H, -CH=), 5.76 (m, 1H, -CH=), 6.19 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH). MS (m / z): 523 (M+H + )。

[0141] Example 2 Preparation of 10-O-(methoxalylallyl)ginkgolide B

[0142]

[0143] Ginkgolide B 300 mg (0.71 mmol) was dissolved in 30 mL THF, 4-bromo-2-butenoic acid methyl ester 251 mg (1.4 mmol), KI 232 mg (1.4 mmol), potassium carbonate 434 mg (3.1 mmol) were added successively, heated to reflux and stirred for 2 hours, TLC tracking to the substrate was basically disappeared, after the reaction was completed, the potassium carbonate was removed by filtration, the mother liquor was concentrated, the residue was column chromatographed, and 192 mg of a light yellow solid was obtained by filtration and drying, with a yield of 52.5%. 1 H-NMR (DMSO-d6, 400 MHz): 1.01 (s, 9H, t-Bu), 1.08 (d, 3H, 14-Me), 1.74 (dd, 1H, 8-H), 1.87 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.73 (s, 1H, -OCH3), 4.06 (m, 1H, 1-H), 4.42 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.80 (d, 1H, J = 16 Hz, 10-CH2-), 5.15 (d, 1H, 1-OH), 5.29 (s, 1H, 10-H), 5.33 (d, 1H, 6-H), 5.65 (m, 1H, -CH=), 5.76 (m, 1H, -CH=), 6.19 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH). MS (m / z): 523 (M+H + ).

[0144] Example 3 Preparation of 10-O-(tert-butoxycarbonylmethyl)ginkgolide B

[0145]

[0146] Ginkgolide B 300 mg (0.70 mmol) was dissolved in 30 mL THF, and then tert- butyl bromoacetate 276 mg (1.41 mmol), KI 235 mg (1.41 mmol), potassium carbonate 434 mg (3.2 mmol) were added successively. The mixture was stirred at reflux for 2 hours. TLC tracking showed that the substrate was almost consumed. After the reaction was completed, the potassium carbonate was removed by filtration, and the mother liquor was concentrated. The residue was purified by column chromatography to give the product 220 mg as light yellow needle crystals with a yield of 58%. 1 H-NMR (DMSO-d6, 400 MHz): 0.99 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.42 (s, 9H, -O-t-Bu), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 4.05 (m, 1H, 1-H), 4.28 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.65 (d, 1H, J = 16 Hz, 10-CH2-), 5.13 (d, 1H, 1-OH), 5.25 (s, 1H, 10-H), 5.31 (d, 1H, 6-H), 6.19 (s, 1H, 12-H), 6.50 (s, 1H, 3-OH). MS (m / z): 539 (M+H + )。

[0147] Example 4 Preparation of 10-O-(methoxycarbonylmethoxyethyl)ginkgolide B

[0148]

[0149] Ginkgolide B 300 mg (0.70 mmol) was dissolved in 30 mL THF, and then methyl iodoethoxyacetate 345 mg (1.41 mmol), KI 235 mg (1.41 mmol), potassium carbonate 434 mg (3.2 mmol) were added successively. The mixture was stirred at reflux for 2 hours. TLC tracking showed that the substrate was almost consumed. After the reaction was completed, the potassium carbonate was removed by filtration, and the mother liquor was concentrated. The residue was purified by column chromatography to give the product 219 mg as light yellow needle crystals with a yield of 58%. 1H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.57 (m, 4H, -OCH2CH2O-), 3.92 (s, 3H, -OCH3), 4.05 (m, 1H, 1-H), 4.28 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.65 (d, 1H, J = 16 Hz, 10-CH2-), 5.13 (d, 1H, 1-OH), 5.25 (s, 1H, 10-H), 5.31 (d, 1H, 6-H), 6.19 (s, 1H, 12-H), 6.50 (s, 1H, 3-OH). MS (m / z): 541 (M+H + ).

[0150] Example 5 Preparation of 10-O-(methoxymethylmethoxyethoxyethyl) ginkgolide B

[0151]

[0152] Ginkgolide B 300 mg (0.70 mmol) was dissolved in 30 mL THF, and then methyl bromoethoxyethoxyacetate 360 mg (1.49 mmol), KI 235 mg (1.41 mmol), potassium carbonate 434 mg (3.2 mmol) were added successively. The mixture was stirred under reflux for 2 hours. The reaction was monitored by TLC until the substrate was consumed. After the reaction was completed, the potassium carbonate was removed by filtration, and the mother liquor was concentrated. The residue was purified by column chromatography to give the product 231 mg as light yellow needle crystals, with a yield of 56%. 1H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.52-3.59 (m, 8H, -OCH2CH2O-), 3.92 (s, 3H, -OCH3), 4.05 (m, 1H, 1-H), 4.28 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.65 (d, 1H, J = 16 Hz, 10-CH2-), 5.13 (d, 1H, 1-OH), 5.25 (s, 1H, 10-H), 5.31 (d, 1H, 6-H), 6.19 (s, 1H, 12-H), 6.50 (s, 1H, 3-OH). MS (m / z): 585 (M-H + ).

[0153] Example 6 Preparation of 4-(Ginkgolide B-10-oxo)-2-butenoic acid

[0154]

[0155] The product of Example 2, 500 mg (0.96 mmol) was dissolved in 6 mL of anhydrous methanol, and lithium hydroxide 51 mg (2.0 mmol) was added. The reaction was stirred at reflux until the substrate was consumed, as monitored by TLC. After 3 hours, the reaction was completed. The pH was adjusted to about 4-5 with dilute hydrochloric acid. The methanol was evaporated, and the solid was allowed to settle. The product was filtered to give 393 mg, 80.6% yield. 1 H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 4.06 (m, 1H, 1-H), 4.42 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.80 (d, 1H, J = 16 Hz, 10-CH2-), 5.15 (d, 1H, 1-OH), 5.29 (s, 1H, 10-H), 5.33 (d, 1H, 6-H), 5.65 (m, 1H, -CH=), 5.76 (m, 1H, -CH=), 6.19 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 13.67 (s, 1H, -COOH). MS (m / z): 507 (M-H + ).

[0156] Example 7 Preparation of 2-(Ginkgolide B-10-oxy)acetic acid

[0157]

[0158] The product from Example 3, 1.18 g (2.19 mmol) was dissolved in 10 mL of chloroform and trifluoroacetic acid, 10 mL was added. The reaction was stirred at room temperature and followed by TLC until the substrate was substantially consumed. The reaction was worked up after 2 hours and the solvent was removed under reduced pressure to give a light yellow solid which was dried by filtration to give the product, 1.05 g, in 99% yield. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.71 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 13.70 (s, 1H, -COOH). MS (m / z): 481 (M-H + ).

[0159] Example 8 Preparation of 2-(Ginkgolide B-10-oxyethoxy)acetic acid

[0160]

[0161] The product from Example 4, 200 mg (0.37 mmol) was dissolved in 6 mL of anhydrous methanol and lithium hydroxide, 25 mg (1.0 mmol) was added. The reaction was stirred at reflux and followed by TLC until the substrate was substantially consumed. The reaction was worked up after 3-4 hours by adjusting the pH to about 4-5 with dilute hydrochloric acid. The methanol was evaporated and the solid was allowed to precipitate. The product was filtered to give 174 mg in 91.9% yield. 1H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, IH, 8-H), 1.86 (ddd, IH, 7α-H), 2.13 (dd, IH, 7β-H), 2.79 (q, IH, 14-H), 3.57 (m, 4H, -OCH2CH2O-), 4.05 (m, IH, 1-H), 4.28 (d, IH, J = 16 Hz, 10-CH2-), 4.63 (d, IH, 2-H), 4.65 (d, IH, J = 16 Hz, 10-CH2-), 5.13 (d, IH, 1-OH), 5.25 (s, IH, 10-H), 5.31 (d, IH, 6-H), 6.19 (s, IH, 12-H), 6.50 (s, IH, 3-OH), 12.8 (s, IH, -COOH). MS (m / z): 525 (M-H + ).

[0162] Example 9 Preparation of 2-(Ginkgolide B-10-oxyethoxyethoxy) acetic acid

[0163]

[0164] The product of Example 5, 100 mg (0.17 mmol) was dissolved in 5 mL of anhydrous methanol, and lithium hydroxide 15 mg (0.6 mmol) was added. The reaction was stirred at reflux until the substrate was consumed, as monitored by TLC. The reaction was stopped after 3-4 hours. The pH was adjusted to about 4-5 with dilute hydrochloric acid, and the methanol was evaporated. The solid was allowed to settle, and the product was filtered to give 82 mg in 84.6% yield. 1 H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, IH, 8-H), 1.86 (ddd, IH, 7α-H), 2.13 (dd, IH, 7β-H), 2.79 (q, IH, 14-H), 3.57 (m, 4H, -OCH2CH2O-), 4.05 (m, IH, 1-H), 4.28 (d, IH, J = 16 Hz, 10-CH2-), 4.63 (d, IH, 2-H), 4.65 (d, IH, J = 16 Hz, 10-CH2-), 5.13 (d, IH, 1-OH), 5.25 (s, IH, 10-H), 5.31 (d, IH, 6-H), 6.19 (s, IH, 12-H), 6.50 (s, IH, 3-OH), 13.5 (s, IH, -COOH). MS (m / z): 569 (M-H + ).

[0165] Example 10 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid sodium salt

[0166]

[0167] 2-(Ginkgolide B-10-oxyl) acetic acid (Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and 2 times molar amount of Na2CO3in methanol was added dropwise (46 mg in 5 mL). After stirring at room temperature for 1 hour, an equal volume of absolute ether was added, and the mixture was left overnight. The solid was separated to give the product 185 mg in 89.5% yield. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH). MS (m / z): 481 (M-H + , neg.).

[0168] Example 11 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid ammonium salt

[0169]

[0170] 2-(Ginkgolide B-10-oxyl) acetic acid (Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and 2 times molar amount of Na2CO3in methanol was added dropwise (46 mg in 5 mL). After stirring at room temperature for 1 hour, an equal volume of absolute ether was added, and the mixture was left overnight. The solid was separated to give the product 185 mg in 89.5% yield. 1H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 1.25 (m, 2H, CH2), 1.78 (m, 2H, CH2), 2.03 (m, 2H, CH2), 3.33 (m, 2H, CH2), 3.48 (m, 1H, CH), 6.82 (br-s, 6H, NH). MS (m / z): 481 (M-H + , neg.).

[0171] Example 12 Preparation of 2-(Ginkgolide B-10-oxyl) Acetic Acid Lysine Salt

[0172]

[0173] 2-(Ginkgolide B-10-oxyl) Acetic Acid (Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and an equimolar amount of lysine in methanol was added dropwise. After stirring at room temperature for 2 hours, the solvent was removed by evaporation under reduced pressure, and 10 mL of diethyl ether was added for washing. The filtrate was obtained as a light yellow solid, 255 mg, in a yield of 98.1%. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 1.25 (m, 2H, CH2), 1.78 (m, 2H, CH2), 2.03 (m, 2H, CH2), 3.33 (m, 2H, CH2), 3.48 (m, 1H, CH), 6.82 (br-s, 6H, NH). MS (m / z): 481 (M-H + , neg.).

[0174] Example 13 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid arginine salt

[0175]

[0176] Example 13 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid arginine salt 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 1.28 (m, 2H, CH2), 1.35 (m, 2H, CH2), 1.82 (m, 2H, CH2), 3.49 (m, 1H, CH), 6.32-7.01 (br-s, 7H, N-H). MS (m / z): 481 (M-H + , neg.).

[0177] Example 14 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid histidine salt

[0178]

[0179] Example 14 Preparation of 2-(Ginkgolide B-10-oxyl) acetic acid histidine salt 1H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 2.90-3.16 (m, 2H, CH2), 4.49 (m, 1H, CH), 7.72 (s, 1H, CH), 8.91 (s, 1H, CH), 6.30-7.01 (br-s, 3H, N-H), 10.25 (br-s, 2H, N-H). MS (m / z): 481 (M-H + , neg.).

[0180] Example 15 Preparation of 2-(Ginkgolide B-10-oxyl)acetic acid triethylamine salt

[0181]

[0182] 2-(Ginkgolide B-10-oxyl)acetic acid (Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and a 2-fold molar amount of triethylamine in methanol was added dropwise. After stirring at room temperature for 2 hours, the solvent was removed under reduced pressure, and 10 mL of diethyl ether was added to wash. The filtrate was obtained as a light yellow solid, 184 mg, in a yield of 76.9%. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09

[0183] (d,3H,14-Me),1.70(dd,1H,8-H),1.82(ddd,1H,7α-H),2.13(dd,1H,7β-H),2.80(q,1H,14- H),4.05(d,1H,1-H),4.29(d,1H,J=16Hz,10-CH2-),4.62(d,1H,2-H),4.75(d,1H,J=16Hz,10 -CH2-),5.28(s,1H,10-H),5.32(d,1H,6-H),5.50(s,1H,1-OH),6.18(s,1H,12-H),6.48(s, 1H,3-OH),1.03(t,9H,CH3-CH2),3.05(q,6H,CH3-CH2),9.25(br-s,1H,NH).MS(m / z):481(MH + ,neg.).

[0184] Example 16 Preparation of 2-(ginkgolide β-10-oxy)acetic acid diethylamine salt

[0185]

[0186] 200 mg (0.41 mmol) of 2-(ginkgolide β-10-oxy)acetic acid (Example 7) was dissolved in 2 mL of anhydrous ethanol, and an equimolar amount of diethylamine in methanol was added. After stirring at room temperature for 2 hours, the solvent was removed by vacuum evaporation, and the mixture was washed with 10 mL of diethyl ether. The mixture was then filtered to give 178 mg of a pale yellow solid, with a yield of 78.2%. 1 H-NMR(DMSO-d6,400MHz):1.00(s,9H,t-Bu),1.09(d,3H,14-Me),1.70(dd,1H,8-H),1.82(ddd,1H,7α-H) ,2.13(dd,1H,7β-H),2.80(q,1H,14-H),4.05(d,1H,1-H),4.29(d,1H,J=16Hz,10-CH2-),4.62(d,1H,2-H ),4.75(d,1H,J=16Hz,10-CH2-),5.28(s,1H,10-H),5.32(d,1H,6-H),5.50(s,1H,1-OH),6.18(s,1H,12- H),6.48(s,1H,3-OH),1.05(t,6H,CH3-CH2),3.07(q,4H,CH3-CH2),8.27(br-s,2H,NH).MS(m / z):481(MH + ,neg.).

[0187] Example 17 Preparation of 2-(Ginkgolide B-10-oxyl)acetic acid n-pentylamine salt

[0188]

[0189] Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and a 3-fold molar amount of n-pentylamine in methanol was added dropwise. After stirring at room temperature for 2 hours, the solvent was evaporated under reduced pressure, 10 mL of diethyl ether was added, and the mixture was filtered to give a light yellow solid, 193 mg, in a yield of 82.7%. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09

[0190] (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 1.05 (m, 3H, CH3-CH2), 1.25-1.30 (m, 4H, -CH2-CH2), 2.05 (m, 2H, -CH2), 3.37 (m, 2H, -CH2), 7.27-7.55 (br-s, 3H, N-H). MS (m / z): 481 (M-H + , neg.).

[0191] Example 18 Preparation of 2-(Ginkgolide B-10-oxyl)acetic acid pyrrolidine salt

[0192]

[0193] Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, and a 3-fold molar amount of n-pentylamine in methanol was added dropwise. After stirring at room temperature for 2 hours, the solvent was evaporated under reduced pressure, 10 mL of diethyl ether was added, and the mixture was filtered to give a light yellow solid, 193 mg, in a yield of 82.7%. 1H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 2.02 (m, 4H, -CH2), 3.17 (m, 4H, -CH2), 7.27-7.55 (br-s, 2H, N-H). MS (m / z): 481 (M-H) + , neg.).

[0194] Example 19 Preparation of 2-(Ginkgolide B-10-oxyl)acetic acid morpholine salt

[0195]

[0196] Example 19 Preparation of 2-(Ginkgolide B-10-oxyl)acetic acid morpholine salt 2-(Ginkgolide B-10-oxyl)acetic acid (Example 7) 200 mg (0.41 mmol) was dissolved in 2 mL of absolute ethanol, 1.5 times molar amount of methanol solution of morpholine was added dropwise, after stirring at room temperature for 2 hours, the solvent was evaporated under reduced pressure, washed with 10 mL of diethyl ether, filtered to obtain a light yellow solid 202 mg, yield 86.6%. 1 H-NMR (DMSO-d6, 400 MHz): 1.00 (s, 9H, t-Bu), 1.09

[0197] (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.82 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.80 (q, 1H, 14-H), 4.05 (d, 1H, 1-H), 4.29 (d, 1H, J = 16 Hz, 10-CH2-), 4.62 (d, 1H, 2-H), 4.75 (d, 1H, J = 16 Hz, 10-CH2-), 5.28 (s, 1H, 10-H), 5.32 (d, 1H, 6-H), 5.50 (s, 1H, 1-OH), 6.18 (s, 1H, 12-H), 6.48 (s, 1H, 3-OH), 3.52 (m, 4H, -CH2), 4.20 (m, 4H, -CH2), 7.28-7.35 (br-s, 2H, N-H). MS (m / z): 481 (M-H+ , neg.).

[0198] Example 20 Preparation of 2-(Ginkgolide B-10-oxyethoxy)acetic acid sodium salt

[0199]

[0200] Example 8) 250 mg (0.48 mmol) was dissolved in 5 mL of absolute ethanol, 1.5 times molar amount of N-methyl-D-glucamine in methanol was added dropwise, after stirring at room temperature for 2 hours, the solvent was evaporated under reduced pressure, 20 mL of diethyl ether was added for washing, and a light yellow solid was obtained by filtration, 245 mg, yield 70.8%. 1 H-NMR (DMSO-d6, 400 MHz): 0.98 (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.57 (m, 4H, -OCH2CH2O-), 4.05 (m, 1H, 1-H), 4.30 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.63 (d, 1H, J = 16 Hz, 10-CH2-), 5.13 (d, 1H, 1-OH), 5.25 (s, 1H, 10-H), 5.31 (d, 1H, 6-H), 6.19 (s, 1H, 12-H), 6.50 (s, 1H, 3-OH). MS (m / z): 525 (M-H + , neg.).

[0201] Example 21 Preparation of 2-(Ginkgolide B-10-oxyethoxy)acetic acid N-methyl-D-glucamine salt

[0202]

[0203] Example 8) 250 mg (0.48 mmol) was dissolved in 5 mL of absolute ethanol, 1.5 times molar amount of N-methyl-D-glucamine in methanol was added dropwise, after stirring at room temperature for 2 hours, the solvent was evaporated under reduced pressure, 20 mL of diethyl ether was added for washing, and a light yellow solid was obtained by filtration, 245 mg, yield 70.8%. 1 H-NMR (DMSO-d6, 400 MHz): 0.98

[0204] (s, 9H, t-Bu), 1.09 (d, 3H, 14-Me), 1.70 (dd, 1H, 8-H), 1.86 (ddd, 1H, 7α-H), 2.13 (dd, 1H, 7β-H), 2.79 (q, 1H, 14-H), 3.57 (m, 4H, -OCH2CH2O-), 4.05 (m, 1H, 1-H), 4.28 (d, 1H, J = 16 Hz, 10-CH2-), 4.63 (d, 1H, 2-H), 4.65 (d, 1H, J = 16 Hz, 10-CH2-), 5.13 (d, 1H, 1-OH), 5.25 (s, 1H, 10-H), 5.31 (d, 1H, 6-H), 6.19 (s, 1H, 12-H), 6.50 (s, 1H, 3-OH), 2.85 (s, 3H, CH3), 3.35-3.39 (m, 9H, CH, OH), 3.58-3.62 (m, 4H, CH2*2), 8.89 (br-s, 2H, NH). MS (m / z): 525 (M-H + ).

[0205] Example 22 Evaluation of in vitro anti-platelet aggregation activity

[0206] Experimental method: New Zealand white rabbits were anesthetized with sodium pentobarbital (2%, 2 ml / kg), and about 40 ml of blood was taken from the abdominal aorta, centrifuged at 1000 rpm / min x 10 min to obtain the supernatant as platelet-rich serum (PRP), and the remaining blood was centrifuged at 4000 rpm / min x 10 min to obtain the supernatant as platelet-poor serum (PPP); blank control group: 100 ul PRP + 100 ul normal saline; drug group: 100 ul PRP + 100 ul drug dissolved in 0.05% DMSO (ginkgolide B as positive control group), the final concentration of the drug was 1 μM. After incubation for 5 min, 2 ul PAF (10 x final concentration of 145 nM) was added; platelet aggregation rates were tested by platelet aggregometer, each sample was repeated five times, the obtained aggregation rates were divided by the average value of the blank control group to obtain the corrected value for comparison;

[0207] The results showed that each example compound showed obvious inhibition effect on platelet aggregation, taking the blank as the standard 1, the platelet aggregation rate of ginkgolide B at a concentration of 1 μM was 63.42%, and the platelet aggregation rates of each example compound were lower than that of ginkgolide B. The specific data are shown in Table 1 and Figure 1 The compounds with more significant activity are Example 2, 7, 12, 13, 14, 17 and 21.

[0208] Table 1. In vitro anti-platelet aggregation activity test results of Examples 1-21

[0209]

[0210] Example 23 Evaluation of Anti-coagulation Activity in vivo in SD Rats

[0211] Method: The rats were randomly divided into groups, 10 rats in each group. After weighing, each group was given the corresponding test drug (50 mg / kg) by gavage, once a day for 3 consecutive days. One hour after the last administration, the rats were anesthetized, and 8 ml of blood was taken from the femoral artery. The whole blood was anticoagulated with 3.8% sodium citrate, and then centrifuged at 800 r / min for 10 min. The upper plasma obtained at this speed was platelet-rich plasma (PRP). After the PRP was aspirated, the remaining blood was centrifuged again at 3000 r / min for 10 min. The plasma obtained at this speed was platelet-poor plasma (PPP).

[0212] PRP 150ul from each group was aspirated and added to a 96-well plate (zeroed with PPP), and ADP inducer was added. The absorbance value A1 was measured at 650 nm wavelength on the microplate reader, which was the absorbance value of each group at 0 min. The shaking mode of the microplate reader was opened, and the absorbance value A2 was detected again after 5 min, which was the absorbance value of each group at 5 min. Since the addition of ADP would cause platelet aggregation and change the light permeability, the maximum aggregation rate (PAGM) of platelets within 5 min was measured based on this principle.

[0213] Platelet maximum aggregation rate (PAGM) = (A1-A2) / A1 x 100

[0214] The results are shown in Table 2 and Figure 2 Each example showed obvious anti-aggregation activity on platelet aggregation in SD rats. Except for Example 10, the activity was higher than that of ginkgolide B. The in vivo activity of Examples 12, 13, and 14 was particularly significant, and could be clinically applied to the treatment of anti-coagulation and thrombosis and related diseases.

[0215] Table 2. In vivo anti-platelet aggregation activity test results of Examples 2, 7, 10, 12, 13, 14, 17, and 21 (n=10)

[0216]

[0217]

[0218] Example 24 Evaluation of Anti-acute Cerebral Ischemia Activity

[0219] Experimental method: SD rats were weighed and randomly divided into groups, 10 rats in each group. Anesthesia was performed by intraperitoneal injection of 15% chloral hydrate 300 mg / kg. The rats were fixed on the rat operating table in left lateral position. The left temporal and facial skin was shaved, and after sterilization with 75% ethanol, the skin was incised between the left eye and the left ear. The temporal muscle and masseter muscle were bluntly separated to expose the temporal bone lamina. Under the operating microscope, a 2 mm x 2 mm bone window was drilled 1 mm from the junction of the temporal bone and squamous temporal bone near the oral side. The skull was pried open with a pry bar. At this time, a relatively straight and less branched blood vessel can be seen through the dura mater, which is the middle cerebral artery. The blood flow was completely blocked by burning with a bipolar electrocoagulation forceps between 1 mm of the olfactory tract and the inferior cerebral vein. The temporal muscle and skin were sutured in turn, and then the rats were given intragastric administration (50 mg / kg). After the rats woke up, they were put back into the cage for further feeding for 24 h.

[0220] The rats were anesthetized again by intraperitoneal injection of 15% chloral hydrate 300 mg / kg. The brain was removed by decapitation, and the olfactory bulb, cerebellum and brainstem were removed. The brain was frozen in a-20℃ refrigerator for about 15 min, and then divided into 7 pieces. The brain slices were stained in 1% TTC staining solution (37℃, avoid light, incubate for about 5-10 min). The infarct area of the stained brain slices was white, and the non-infarct area was rose red. After staining, the brain slices were moved to 10% formaldehyde solution and stored in the dark for 24 h. Finally, a digital camera was used to take pictures, and the image analysis software (ImageJ, version: 1.4.3.67) was used to measure the area of the infarct area and the non-infarct area on the front and back surfaces, and calculate the percentage of the infarct area to the total area of the infarct hemisphere.

[0221] The experimental results are shown in Table 3 and Figure 3 The results show that compared with the model group, the compounds of each example can significantly reduce the proportion of half-brain infarct area of rats. Except for example 10, the proportion of half-brain infarct area of the other groups is less than that of ginkgolide B group. The activities of examples 12, 13 and 14 are particularly significant, and can be clinically applied to the treatment of cerebral ischemia and cerebral ischemia related diseases.

[0222] Table 3. Activity evaluation results of examples 2, 7, 10, 12, 13, 14 and 21 for reducing the proportion of half-brain infarct area of SD rats (n=10)

[0223] Compound Mean value of proportion of half brain infarction area (%) Standard deviation Model group 31.91 1.66 Ginkgo biloba extract 26.12 6.04 Example 2 21.56 4.35 Example 7 25.14 4.56 Example 10 29.61 3.00 Example 12 18.83 5.75 Example 13 20.14 5.27 Example 14 17.98 4.76 Example 17 25.31 5.13 Example 21 22.66 3.67

[0224] Although the specific embodiments of the present application are described above, those skilled in the art should understand that these are only illustrative, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present application. Therefore, the protection scope of the present application is defined by the appended claims.

Claims

1. A compound, characterized in that, The compound is selected from any of the following compounds: ， 。 2. A pharmaceutical composition, characterized in that, It contains a therapeutically effective dose of the compound as described in claim 1.

3. Use of a compound as described in claim 1, or a pharmaceutical composition as described in claim 2, in the preparation of a medicament for the prevention and treatment of ischemic stroke, thrombosis, angina pectoris, myocardial infarction, and inflammatory or asthma-related diseases associated with platelet-activating factor.

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