CN108191722B

CN108191722B - P2Y12 receptor antagonist with halogenated diphenyl thioacetic acid structure and application thereof

Info

Publication number: CN108191722B
Application number: CN201810050950.3A
Authority: CN
Inventors: 曾华仙
Original assignee: Tianjin Xiaoxin Pharmaceutical Technology Co ltd
Current assignee: Tianjin Xiaoxin Pharmaceutical Technology Co ltd
Priority date: 2018-01-18
Filing date: 2018-01-18
Publication date: 2021-06-29
Anticipated expiration: 2038-01-18
Also published as: CN108191722A

Abstract

The present invention relates to the field of medicaments associated with cardiovascular diseases. Specifically, the invention relates to a P2Y12 receptor antagonist containing a halogenated diphenyl thioacetic acid structure, a preparation method thereof and application thereof in preparing medicaments for treating cardiovascular diseases, especially thromboembolic diseases.

Description

P2Y12 receptor antagonist with halogenated diphenyl thioacetic acid structure and application thereof

Technical Field

The invention relates to the field of medicaments for treating vascular diseases. In particular to a P2Y12 receptor antagonist containing halogenated diphenyl thioacetic acid structure, which has the treatment effect on vascular diseases, in particular to thromboembolic diseases, a preparation method and the application thereof in pharmacy.

Background

Medical complications associated with the development of thrombosis represent a major cause of death. Some examples of pathologies associated with the development of thrombosis include acute myocardial infarction, unstable angina and chronic stable angina, transient ischemic attacks, cerebrovascular accidents, peripheral vascular diseases, preeclampsia and eclampsia, deep vein thrombosis, embolism (cerebral embolism, pulmonary embolism, coronary artery embolism, renal embolism, etc.), disseminated intravascular coagulation or thrombotic thrombocytopenic purpura. There is still a risk of thrombotic and restenotic complications during and after invasive surgery, such as angioplasty, carotid endarterectomy, aortic coronary artery bypass grafting or placement of stents or endovascular prostheses.

Arterial thrombosis may occur following injury to the vessel wall or rupture of an atheromatous plaque. Platelets play an essential role in the formation of these thrombi. Platelets can be activated by: mediators released by circulating cells in the bloodstream or by damaging endothelial cells present along the walls of blood vessels, or by thrombotic molecules of the subendothelial matrix (e.g., collagen) exposed during vascular injury. In addition, platelets can also be activated under conditions of blood flow with high shear stress, as observed in stenotic vessels. Upon activation, the circulating platelets adhere to and accumulate at the site of the vascular injury, forming a thrombus. In this process, the thrombus created in the blood vessel is sufficiently voluminous for the blood flow to become partially or completely occluded.

In veins, thrombi can also form where there is an obstruction or slow blood flow. Due to the nature of these venous thrombi, they can produce emboli that migrate within the vascular system. These emboli can thus block blood flow in more distant vessels, such as the pulmonary or coronary arteries.

Numerous studies have demonstrated that adenosine 5' -diphosphate (ADP) is the major mediator of platelet activation and aggregation, playing a crucial role in the initiation and progression of thrombosis (Maffrand et al, Thromb. Haemostas.,1988,59, 225-230). ADP is released into the circulation by damaged erythrocytes and endothelial cells of the atherosclerotic wall, and more specifically is secreted by activated platelets where ADP is stored in very high concentrations in dense granules. ADP-induced platelet aggregation is triggered by its binding to two specific purinergic receptors P2Y1 and P2Y12 expressed in the cell membrane of human platelets. The P2Y1 receptor, in combination with PLC β stimulation by G α q, is responsible for mobilization of internal calcium stores, alteration of platelet shape and transient aggregation on ADP. The P2Y12, in combination with inhibition of adenylate cyclase by G α i2 and activation of PI-3 kinase, is responsible for amplification of the response and stabilization of aggregation. The use of P2Y 1-/-transgenic mice (Gachet et al, J. Clin. invest.,1999,104,1731-1737) and P2Y 12-/-mice (Conley et al, Nature,2001,409, 202-173207) demonstrated the importance of the two receptors in thrombus development in vivo. In humans, P2Y12 gene deficiency has been described to be associated with a bleeding phenotype and a significant decline in ADP-induced platelet aggregation. The use of clopidogrel in human clinical practice has demonstrated that blockade of the P2Y12 receptor by antagonists represents a key therapeutic strategy for the treatment of cardiovascular diseases. Clopidogrel is a prodrug of the thienopyridine family, whose active metabolite covalently binds to the P2Y12 receptor, resulting in irreversible inhibition of platelet activity in vivo (Savi et al, biochem. biophysis. res. commun.,2001,283,379-383), which drug has shown its efficacy in several clinical trials, i.e. reducing the risk of cardiovascular accidents in patients at risk.

The invention discloses P2Y12 receptor antagonists containing halogenated diphenyl thioacetic acid structures, which can be used for preparing medicaments for treating cardiovascular diseases, in particular thromboembolic diseases.

Disclosure of Invention

It is an object of the present invention to provide a P2Y12 receptor antagonist with good activity of the general formula I.

It is another object of the present invention to provide a process for the preparation of compounds having the general formula I.

The present disclosure will now be described in detail for the purpose of the invention.

The compounds of the present invention having the general formula I have the following structural formula:

wherein X is selected from F, Cl substituent.

The compound of the general formula I can be synthesized by the following route:

reacting the compound II with triphenylphosphine to obtain a salt III; treating the compound III with n-butyllithium to obtain the corresponding phosphine ylide IV, and reacting the phosphine ylide IV with the compound V under the condition of heating to obtain a compound VI; reacting the compound VI with bromine to obtain a dibromo-compound VII; reacting the compound VII with mercaptoacetic acid under an alkaline condition to obtain a compound I; x is as defined above.

The compound of the general formula I has antagonistic action of a P2Y12 receptor, and can be used as an effective component for preparing medicaments for treating cardiovascular diseases, particularly thromboembolic diseases. The activity of the compounds of the general formula I of the invention is verified by in vitro inhibition tests of human platelet aggregation.

The compounds of formula I of the present invention are effective over a relatively wide dosage range. For example, the daily dosage may be in the range of about 1mg to about 700mg per person, divided into one or more administrations. The actual dosage of the compounds of formula I to be administered according to the invention can be determined by the physician in the light of the relevant circumstances.

Detailed Description

The present invention will be further described with reference to the following examples. It should be noted that the following examples are only for illustration and are not intended to limit the present invention. Variations of the teachings of the present invention may be made by those skilled in the art without departing from the scope of the claims of the present application.

EXAMPLE 1 Synthesis of Compound I-1

Step 1 Synthesis of Compound III-1

Compound II-1(1.63g,10mmol) and triphenylphosphine (2.62g,10mmol) were dissolved in 30mL of chloroform, and refluxed overnight with stirring to precipitate a large amount of crystals. The reaction mixture was cooled, the solid collected by suction filtration and dried in vacuo at room temperature to give compound III-1, a white crystalline solid, ESI-MS, M/z ═ 345([ M-Br)]⁺)。

Step 2 Synthesis of Compound VI-1

Compound III-1(3.40g,8mmol) was dissolved in 50mL of dry THF, stirred, cooled to-20 ℃ under a nitrogen atmosphere, then 1.6M n-BuLi in n-hexane (5mL,8mmol) was slowly added dropwise via syringe, and after the addition was complete, the reaction mixture was stirred at that temperature for 1 hour. A solution of benzophenone (V-1, 1.46g,8mmol) in 5mL of dry THF was then added slowly dropwise with a syringe, and after the addition was complete, the reaction mixture was stirred at that temperature for half an hour, then warmed to room temperature and stirred for 1 hour, and finally stirred under reflux overnight. The reaction mixture was carefully poured into 300mL of ice-water, stirred, and then 50mL of X3 CH was used₂Cl₂Extraction, combining extract phases, washing with brine and drying with anhydrous sodium sulfate. The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound VI-1, ESI-MS, M/z ═ 249([ M + H)]⁺) A white solid.

Step 3. Synthesis of Compound VII-1

Compound VI-1(0.99g,4mmol) is dissolved in 10mL CH₂Cl₂In the process, stirring and slowly dripping Br₂(0.80g,5mmol) and 1mL CH₂Cl₂The prepared solution was added dropwise, and the reaction mixture was stirred at room temperature overnight and checked by TLC to find completion. To the reaction mixture was added 30mL of diethyl ether and 20mL of n-hexane, stirred at room temperature for 1 hour, filtered, the solid was collected and dried under vacuum at room temperature to give compound VII-1, ESI-MS, M/z ═ 409([ M + H) ([ M + H ])]⁺)。

Step 4. Synthesis of Compound I-1

Mercaptoacetic acid (0.28g,3mmol) was dissolved in 10mL of ethanol, stirred, and 30% NaOH solution (1mL) was slowly added dropwise, followed by the addition of Compound VII-1(0.82g,2mmol), the reaction mixture was refluxed for 24 hours under nitrogen atmosphere and checked by TLC to find completion. The reaction mixture was carefully poured into 200mL of ice water, stirred, adjusted to pH 2-3 with concentrated HCl, and 50mL of X3 CH was used₂Cl₂Extraction, combining extract phases, washing with brine and drying with anhydrous sodium sulfate. The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound I-1, ESI-MS, M/z ═ 337([ M-H)]^-) A white solid.

EXAMPLE 2 Synthesis of Compound I-2

Step 1 Synthesis of Compound III-2

Compound II-2(1.77g,10mmol) and triphenylphosphine (2.62g,10mmol) were dissolved in 30mL of chloroform, and refluxed overnight with stirring to precipitate a large amount of crystals. The reaction mixture was cooled, the solid collected by suction filtration and dried in vacuo at room temperature to give compound III-2 as a white crystalline solid, ESI-MS, M/z 359([ M-Br)]⁺)。

Step 2 Synthesis of Compound VI-2

Compound III-2(3.51g,8mmol) was dissolved in 50mL of dry THF, stirred, cooled to-20 ℃ under a nitrogen atmosphere, then 1.6M n-BuLi in n-hexane (5mL,8mmol) was slowly added dropwise via syringe, and after the addition was complete, the reaction mixture was stirred at that temperature for 1 hour. A solution of V-2(1.74g,8mmol) in 5mL of dry THF was again added slowly dropwise via syringe, and after the addition was complete, the reaction mixture was stirred at this temperature for half an hour, then warmed to room temperature and stirred for 1 hour, and finally stirred under reflux overnight. The reaction mixture was carefully poured into 300mL of ice-water, stirred, and then 50mL of X3 CH was used₂Cl₂Extracting, combining the extract phase,washed with brine and dried over anhydrous sodium sulfate. The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound VI-2, ESI-MS, M/z ═ 299([ M + H)]⁺) A white solid.

Step 3. Synthesis of Compound VII-2

Compound VI-2(1.19g,4mmol) is dissolved in 10mL CH₂Cl₂In the process, stirring and slowly dripping Br₂(0.80g,5mmol) and 1mL CH₂Cl₂The prepared solution was added dropwise, and the reaction mixture was stirred at room temperature overnight and checked by TLC to find completion. To the reaction mixture was added 30mL of diethyl ether and 20mL of n-hexane, stirred at room temperature for 1 hour, filtered, the solid was collected by suction, and dried under vacuum at room temperature to give compound VII-2, ESI-MS, M/z ═ 459([ M + H ═ 459)]⁺)。

Step 4. Synthesis of Compound I-2

Mercaptoacetic acid (0.28g,3mmol) was dissolved in 10mL of ethanol, stirred, and 30% NaOH solution (1mL) was slowly added dropwise, followed by the addition of compound VII-2(0.92g,2mmol), the reaction mixture was refluxed for 24 hours under nitrogen atmosphere and checked by TLC to find completion. The reaction mixture was carefully poured into 200mL of ice water, stirred, adjusted to pH 2-3 with concentrated HCl, and 50mL of X3 CH was used₂Cl₂Extraction, combining extract phases, washing with brine and drying with anhydrous sodium sulfate. The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound I-2 as a white solid, ESI-MS, M/z 387([ M-H)]^-)。

EXAMPLE 3 Synthesis of Compound I-3

Step 1 Synthesis of Compound III-2

Compound II-2(1.77g,10mmol) and triphenylphosphine (2.62g,10mmol) were dissolved in 30mL of chloroform, and refluxed overnight with stirring to precipitate a large amount of crystals. Cooling the reaction mixture, filtering, collecting solid, and vacuum drying at room temperature to obtainCompound III-2, white crystalline solid, ESI-MS, M/z 359([ M-Br)]⁺)。

Step 2 Synthesis of Compound VI-3

Compound III-2(3.51g,8mmol) was dissolved in 50mL of dry THF, stirred, cooled to-20 ℃ under a nitrogen atmosphere, then 1.6M n-BuLi in n-hexane (5mL,8mmol) was slowly added dropwise via syringe, and after the addition was complete, the reaction mixture was stirred at that temperature for 1 hour. A solution of V-3(2.01g,8mmol) in 5mL of dry THF was again slowly added dropwise via syringe, and after completion of the addition, the reaction mixture was stirred at this temperature for half an hour, then warmed to room temperature and stirred for 1 hour, and finally stirred under reflux overnight. The reaction mixture was carefully poured into 300mL of ice-water, stirred, and then 50mL of X3 CH was used₂Cl₂Extraction, combining extract phases, washing with brine and drying with anhydrous sodium sulfate. The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound VI-3, ESI-MS, M/z 332([ M + H)]⁺) A white solid.

Step 3. Synthesis of Compound VII-3

Compound VI-3(1.32g,4mmol) is dissolved in 10mL CH₂Cl₂In the process, stirring and slowly dripping Br₂(0.80g,5mmol) and 1mL CH₂Cl₂The prepared solution was added dropwise, and the reaction mixture was stirred at room temperature overnight and checked by TLC to find completion. To the reaction mixture was added 30mL of diethyl ether and 20mL of n-hexane, stirred at room temperature for 1 hour, filtered with suction, the solid was collected and dried under vacuum at room temperature to give compound VII-3, ESI-MS, M/z ═ 492([ M + H)]⁺)。

Step 4. Synthesis of Compound I-3

Mercaptoacetic acid (0.28g,3mmol) was dissolved in 10mL of ethanol, stirred, and 30% NaOH solution (1mL) was slowly added dropwise, followed by the addition of compound VII-3(0.98g,2mmol), the reaction mixture was refluxed for 24 hours under nitrogen atmosphere and checked by TLC to find completion. The reaction mixture was carefully poured into 200mL of ice water, stirred, adjusted to pH 2-3 with concentrated HCl, and 50mL of X3 CH was used₂Cl₂Extraction, combining extract phases, washing with brine and drying with anhydrous sodium sulfate.The drying agent was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator and the residue was purified by silica gel column chromatography to give compound I-3 as a white solid, ESI-MS, M/z ═ 420([ M-H)]^-)。

EXAMPLE 4 in vitro inhibition of human blood platelets by Compounds

Blood was collected from healthy volunteers using a 20mL syringe containing 2mL of buffered sodium citrate. The blood was transferred to polypropylene tubes and centrifuged (100g) at room temperature for 5 minutes (brake without centrifuge). Platelet Rich Plasma (PRP) floating on the surface was then collected, diluted, and subjected to platelet counting before being used for aggregation measurements.

Measurement of platelet aggregation was performed in a glass tube at 37 ℃ (platelet aggregometer). mu.L of test compound (100-fold more concentrated DMSO solution than the desired final concentration) was mixed with 392. mu.L of freshly prepared PRP and incubated for 1 min with stirring. Then 4. mu.L of 250. mu.M ADP solution was added to the mixture. The measurement of aggregation was monitored by recording the optical density change according to the method of g.v.r.born (Born, Nature,1962,194,927) for 6 to 8 minutes with constant stirring. Aggregate amplitude calculations, expressed as height, were used and expressed as percent inhibition. IC (for platelet aggregation inhibition) of the Compounds of the invention₅₀As shown in the table below.

Compound (I)	IC₅₀(nM)
		Compound I-1	761
Compound I-2	209
		Compound I-3	344

From the results in the table, the compound of the invention has strong antagonistic effect on P2Y12, and can be used for preparing medicines for treating cardiovascular diseases, especially thromboembolic diseases.

Claims

1. A compound having the structure of the general formula I,

wherein X is selected from F, Cl substituent.

2. A method of synthesizing the compound of claim 1:

reacting the compound II with triphenylphosphine to obtain a salt III; treating the compound III with n-butyllithium to obtain corresponding phosphine ylide IV, and reacting the compound IV with the compound V under the condition of heating to obtain a compound VI; reacting the compound VI with bromine to obtain a dibromo-compound VII; reacting the compound VII with mercaptoacetic acid under an alkaline condition to obtain a compound I; x is as defined in claim 1.

3. The use of a compound according to claim 1 for the manufacture of a medicament for the treatment of thromboembolic disorders.