MENTHOL class P2Y12 receptor antagonist, preparation method and its usageTechnical field
The present invention relates to the drug world for the treatment of cardiovascular disease.In particular it relates to outstanding to cardiovascular diseaseWhich is that thrombotic disease has P2Y12 receptor antagonist of the medicative class containing MENTHOL structure, its preparation sideMethod, and the purposes in pharmacy.
Background technology
The medical complication relevant with there are thrombosiss represents a kind of main cause of death.Some and development thrombosis shapeInclude that acute myocardial infarction, unstable angina pectoriss and chronic stable angina pectoris, transience lack into relevant pathology exampleBlood outbreak, cerebrovas-cularaccident, peripheral vascular disease, preeclampsia and eclamposia, dvt formed, thromboembolism (cerebral embolism,Pulmonary infarction, coronary thrombosiss, renal infarction etc.), disseminated inravascular coagulation or thrombotic thrombocytopenic purpura.InvadingStill there are the danger that thrombosiss and restenosiss complication occur, the invasive surgical operation during and after entering property surgical operationSuch as angioplasty, carotid endarterectomy, aorto-coronary bypass grafting or support or the peace of vascular endoprosthesesPut.
Artery thrombosis can occur after vascular damaged or atheromatous plaque rupture.Platelet is in these thrombosisNecessary effect is played in formation.Platelet can pass through following substance activating：In blood flow circulating cells or along blood vessel wall presentThe amboceptor that discharged of damaging endotheliocyte, or during blood vessel injury exposed sub-endothelial matrix (such as collagen) bloodBolt forms molecule.Additionally, platelet can also be under the blood flow conditions with shearing force as observed by narrow blood vesselAs activate.After activation, the circulation platelet adhesion is simultaneously accumulated at blood vessel injury, forms thrombosis.In this process, bloodThrombosis produced by pipe are that volume is sufficiently large to blood flow, so which is partially or completely blocked.
In vein, thrombosis can also be located to be formed in obstruction or blood flow slowly.Due to these venothrombotic properties, which can produceThe embolus moved in vascular system.These emboluses thus the blood flow in more remote blood vessel can be blocked, the blood vessel such as lungTremulous pulse or coronary artery.
Many research principal mediators that verified 5'- adenosine diphosphate (ADP)s (ADP) are platelet activation and assemble, in thrombosisPlay in the startup of formation and progress decisive role (Maffrand et al., Thromb.Haemostas., 1988,59,225-230).ADP is discharged in circulation by the endotheliocyte of the erythrocyte that damages and atherosclerosiss wall, it is more specific andSpeech, by secreted by the activation platelet with compacted grains in place of very high concentration stores ADP.The platelet aggregation of ADP- inductionsTriggered by which and in the combination of two species specificity purinergic receptor P2Y1 and P2Y12 of the endoglin expression of human blood platelets.InstituteP2Y1 receptors are stated, is stimulated with the PLC β of Jing G α q and combine, be responsible for mobilization, the change of platelet shape that internal calcium stores and in ADPOn moment aggregation.The P2Y12, the activation of suppression and PI-3 kinases with the adenyl cyclase of Jing G α i2 combine, and are responsible forThe stabilisation of the amplification and aggregation of response.P2Y1-/- transgenic mice (Gachet et al., J.Clin.Invest., 1999,104,1731-1737) prove with using for P2Y12-/- mice (Conley et al., Nature, 2001,409,202-207)Described two receptors thrombosis developing importance in vivo.In the mankind, P2Y12 genetic flaws and bleeding table are had been described aboveType is relevant with the notable decline of the platelet aggregation that ADP- is induced.Human clinical practice used in clopidogrel it has been proved thatThe critical therapeutic strategy for the treatment of cardiovascular disease is represent by Antagonist block P2Y12 receptors.Clopidogrel is thienopyridineThe prodrug of family, its active metabolite are covalently bond to P2Y12 receptors, and cause internal biologically active pdgf can not retroactive inhibition(Savi et al., Biochem.Biophys.Res.Commun., 2001,283,379-383), the medicine is in some clinical examinationsMiddle its efficiency of display is tested, that is, reduces adventurous patient's generation cardiovascular unexpectedly dangerous.
The invention discloses a class contains the P2Y12 receptor antagonists of MENTHOL structure, these compounds can be used to prepareThe medicine for the treatment of cardiovascular disease especially thrombotic disease.
The content of the invention
It is an object of the present invention to provide a kind of P2Y12 receptor antagonists of the excellent activity with formula I.
It is a further object to provide preparing the method with compounds of formula I.
It is also another object of the present invention to provide containing compounds of formula I as effective ingredient and its in treatment cardiovascularDisease application especially in terms of thrombotic disease.
Present invention is specifically described in conjunction with the purpose of the present invention.
The present invention has following structural formula with compounds of formula I：
Wherein, R is selected from H, C1-C8Alkyl, C3-C8Cycloalkyl.
It is preferred that below general formula I,
Wherein, R is selected from H, C1-C3Alkyl, C3-C6Cycloalkyl.
More preferably compounds of formula I has following structure,
Compound of Formula I of the present invention can be synthesized by following route：
Reacting by heating generates compound IV in the presence of a base for compound II and compound III；Compound IV reducing agents are alsoOriginal obtains compound V；Compound V is reacted with chloracetyl chloride in the presence of a base, obtains compound VI；Compound VI is in the presence of a baseReact with sulfhydryl compound VII, obtain corresponding I；R is defined as described above.
Compound of Formula I of the present invention has the antagonism of P2Y12 receptors, can be used to prepare the heart as effective ingredientAngiopathy especially thrombotic disease medicine.The activity of compound of Formula I of the present invention is by external peopleThe inhibition test of blood platelet aggregation is verifying.The compound of Formula I of the present invention is effective in comparatively wide dosage range's.The dosage for example taken daily about in the range of 1mg-700mg/ people is divided into and being administered once or for several times.The present invention is taken actuallyThe dosage of compound of Formula I can be determined according to relevant situation by doctor.
Specific embodiment
With reference to embodiment, the present invention is further illustrated.It should be noted that following embodiments are only forIllustrate, and be not intended to limit the present invention.The various change that those skilled in the art's training centre of the invention is made all shouldWithin the protection domain required by the application claim.
The synthesis of 1 compound I-1 of embodiment
Synthesis 1.56g (10mmol) the compound II of step 1. compound IV is dissolved in 15mL DMF, under ice-water bath coolingStirring, (12mmol, 60%) solid NaH are stirred 30 minutes under room temperature to be dividedly in some parts 0.48g.Add 1.86g (10mmol) 2,4-Dinitro compound III, then stirs 3 hours under room temperature, and TLC shows that reaction is completed.Reactant mixture carefully pours into 200mLIn frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate drying.Sucking filtration is removedDesiccant, filtrate are evaporated on a rotary evaporator, and residue uses silica gel column chromatography purification, obtains compound IV, white solid,ESI-MS, m/z=323 ([M+H]+)。
Synthesis 2.26g (7mmol) the compound IV of step 2. compound V is dissolved in 20mL dehydrated alcohol, stirring, is added0.1g 10%Pd/C, then according to standard operation carries out being hydrogenated with normal temperature and pressure, complete after 12 hours.Reactant mixture is carefulPour in 200mL frozen water, stir, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate to doIt is dry.Sucking filtration removes desiccant, and filtrate is evaporated on a rotary evaporator, and residue uses silica gel column chromatography purification, obtains compoundV, white solid, ESI-MS, m/z=263 ([M+H]+)。
Synthesis 1.31g (5mmol) the compound V of step 3. compound VI is dissolved in the dichloromethane of 15mL dryings, frozen waterBath cooling is lower to stir, and adds 1.52g (15mmol) triethylamine, and then slowly Deca 0.56g (5mmol) chloracetyl chloride and 1mL are dryThe solution that dry dichloromethane is prepared, after completion of dropping, stirring was continued at room temperature overnight for reactant mixture, and TLC shows and reactedInto.Reactant mixture is carefully poured in 200mL frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses successively1% hydrochloric acid and salt water washing, anhydrous sodium sulfate drying.Sucking filtration removes desiccant, and filtrate is evaporated on a rotary evaporator, remainingThing uses silica gel column chromatography purification, obtains compound VI, white-yellowish solid, ESI-MS, m/z=339 ([M+H]+)。
Synthesis 1.02g (3mmol) compound VI, 0.33g (3mmol) benzyl mercaptan VII-1 of step 4. compound I-1 and0.91g (9mmol) triethylamine is dissolved in the dichloromethane of 10mL dryings, and stirring was continued at room temperature overnight for reactant mixture, TLCShow that reaction is completed.Reactant mixture is carefully poured in 200mL frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merges extractionPhase, successively with 1% hydrochloric acid and salt water washing, anhydrous sodium sulfate drying.Sucking filtration removes desiccant, and filtrate is on a rotary evaporatorIt is evaporated, residue uses silica gel column chromatography purification, obtains compound I-1, white solid, ESI-MS, m/z=413 ([M+H]+)。
The synthesis of 2 compound I-2 of embodiment
Synthesis 1.56g (10mmol) the compound II of step 1. compound IV is dissolved in 15mL DMF, under ice-water bath coolingStirring, (12mmol, 60%) solid NaH are stirred 30 minutes under room temperature to be dividedly in some parts 0.48g.Add 1.86g (10mmol) 2,4-Dinitro compound III, then stirs 3 hours under room temperature, and TLC shows that reaction is completed.Reactant mixture carefully pours into 200mLIn frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate drying.Sucking filtration is removedDesiccant, filtrate are evaporated on a rotary evaporator, and residue uses silica gel column chromatography purification, obtains compound IV, white solid,ESI-MS, m/z=323 ([M+H]+)。
Synthesis 2.26g (7mmol) the compound IV of step 2. compound V is dissolved in 20mL dehydrated alcohol, stirring, is added0.1g 10%Pd/C, then according to standard operation carries out being hydrogenated with normal temperature and pressure, complete after 12 hours.Reactant mixture is carefulPour in 200mL frozen water, stir, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses salt water washing, anhydrous sodium sulfate to doIt is dry.Sucking filtration removes desiccant, and filtrate is evaporated on a rotary evaporator, and residue uses silica gel column chromatography purification, obtains compoundV, white solid, ESI-MS, m/z=263 ([M+H]+)。
Synthesis 1.31g (5mmol) the compound V of step 3. compound VI is dissolved in the dichloromethane of 15mL dryings, frozen waterBath cooling is lower to stir, and adds 1.52g (15mmol) triethylamine, and then slowly Deca 0.56g (5mmol) chloracetyl chloride and 1mL are dryThe solution that dry dichloromethane is prepared, after completion of dropping, stirring was continued at room temperature overnight for reactant mixture, and TLC shows and reactedInto.Reactant mixture is carefully poured in 200mL frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merges extraction phase, uses successively1% hydrochloric acid and salt water washing, anhydrous sodium sulfate drying.Sucking filtration removes desiccant, and filtrate is evaporated on a rotary evaporator, remainingThing uses silica gel column chromatography purification, obtains compound VI, white-yellowish solid, ESI-MS, m/z=339 ([M+H]+).Step 4.Synthesis 1.02g (3mmol) compound VI, 0.37g (3mmol) VII-2 and 0.91g (9mmol) triethylamine of compound I-2 is dissolved inIn the dichloromethane that 10mL is dried, stirring was continued at room temperature overnight for reactant mixture, and TLC shows that reaction is completed.Reaction mixingThing is carefully poured in 200mL frozen water, stirring, with 50mL × 3CH2Cl2Extraction, merge extraction phase, successively with 1% hydrochloric acid andSalt water washing, anhydrous sodium sulfate drying.Sucking filtration removes desiccant, and filtrate is evaporated on a rotary evaporator, and residue uses silica gelColumn chromatography purification, obtains compound I-2, white solid, ESI-MS, m/z=427 ([M+H]+)。
Embodiment 3-10
With reference to the method for embodiment 1,2, compound listed in Table is synthesized.
12 Compound ira vitro of embodiment is to the hematoblastic inhibitory action of human blood
Using the 20mL syringes of the sodium citrate buffered containing 2mL, blood is gathered from healthy volunteer.Blood is shiftedInto polypropylene tube, and (100g) 5 minutes (not using the braking of centrifuge) is centrifuged in room temperature.Then collect supernatant richnessThrombocyte plasma (PRP), dilution, and platelet count was carried out before aggregation measurement is used it for.
37 DEG C of measurements (platelet aggregation instrument) for carrying out platelet aggregation in glass tubing.By 4 μ L test compounds (than needingThe DMSO solution of dense 100 times of the final concentration wanted) mix with the PRP of 392 μ L brand-news, and with stirring incubation 1 minute.Then to mixedThe ADP solution of 250 μM of 4 μ L is added in compound.Persistently stir, by the method recording light variable density according to G.V.R.Born(Born, Nature, 1962,194,927), monitor the measurement of aggregation 6 to 8 minutes.Using the aggregation amplitude meter represented with heightResult is calculated, and is represented with suppression percentage.The inhibition on platelet aggregation IC of the compounds of this invention50It is as shown in the table.
| Compound | IC50(nM) |
| I-1 | 734 |
| I-2 | 209 |
| I-3 | 241 |
| I-4 | 793 |
| I-5 | 855 |
| I-6 | 461 |
| I-7 | 689 |
| I-8 | 1057 |
| I-9 | 334 |
| I-10 | 577 |
| I-11 | 1003 |
The compound that the present invention be can be seen that from upper table result has very strong antagonism to P2Y12, can be used as systemThe medicine of standby treatment cardiovascular disease especially thrombotic disease.