阅读说明：本技术 氯吡格雷活性代谢物衍生物、其前药及其制备方法与应用 (Clopidogrel active metabolite derivative, prodrug thereof, preparation method and application thereof ) 是由 杨慧 马宇衡 牧童 布仁 赵岩 于 2020-07-28 设计创作，主要内容包括：本发明公开了氯吡格雷活性代谢物衍生物、其前药及其制备方法与应用,其中所述衍生物具有如式(A)所示的结构式,其能有效应用于抗血小板聚集药物中,且活性高、生物利用度高,起效快,所述前药结构稳定、毒副作用小,所述制备方法立体选择性高,普适性强,过程简单,目标产物产率高,<Image he="369" wi="561" file="DDA0002604822770000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention discloses a clopidogrel active metabolite derivative, a prodrug thereof, a preparation method and an application thereof, wherein the derivative has a structural formula shown in a formula (A), can be effectively applied to platelet aggregation resistant medicines, and has the advantages of high activity, high bioavailability, quick response, stable prodrug structure, small toxic and side effects, high stereoselectivity, strong universality, simple process and high yield of target products,)

1. A clopidogrel active metabolite derivative, a prodrug or a pharmaceutically acceptable salt thereof, wherein the derivative has a structural formula as shown in formula (a):

wherein the content of the first and second substances,

R₁any one selected from a straight chain alkyl group having 1 to 10 carbon atoms, an alkyl group having a main chain having 1 to 10 carbon atoms and containing a branched chain, a hydrocarbon acyl group having 1 to 10 carbon atoms, an aromatic cyclic acyl group and a carbamido group;

R₂any one selected from hydrogen, a straight chain alkyl group having 1 to 10 carbon atoms, and an alkyl group having a main chain with 1 to 10 carbon atoms and containing a branched chain.

2. The clopidogrel active metabolite derivative according to claim 1, characterized in that: the derivative is (Z) -2- ((S) -1- ((S) -1- (2-chlorphenyl) -2-methoxyl-D3-2-oxyethyl) -4- (methylthio) piperidine-3-methylene) acetic acid shown as a structural formula AQ-10:

3. the prodrug of a clopidogrel active metabolite derivative according to claim 1, characterized in that: which is selected from one or more of the following compounds:

methyl-D3 (S) -2- ((S, Z) -4-acetylsulfanyl-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate;

methyl-D3 (S) -2- ((S, Z) -4-isothioureido-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate;

(Z) -2- ((S) -1- (2-chlorophenyl) -2-methoxy-D3-2-oxoethyl) -4- (methylthio) piperidin-3-ylidene) acetic acid;

methyl-D3 (S) -2- ((S, Z) -4-benzoylsulfanyl-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate;

methyl-D3 (S) -2- ((S, Z) -4-phenylacetylthio-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate;

methyl-D3 (S) -2- ((S, Z) -4-isopropenylthio-3- (2-ethoxy-2-oxyethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate;

methyl-D3 (S) -2- ((S, Z) -4-propionylthio-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate.

4. The prodrug of claim 3, wherein: the prodrug has a structural formula shown as a formula (B):

5. a method for preparing a clopidogrel active metabolite derivative, comprising:

by reacting a compound of the formula XIV with a compound S-R₁Or the compound S-R₁The salt is subjected to bimolecular nucleophilic substitution reaction to obtain the derivative,

wherein the content of the first and second substances,

R₁any one selected from a straight chain alkyl group having 1 to 10 carbon atoms, an alkyl group having a main chain having 1 to 10 carbon atoms and containing a branched chain, a hydrocarbon acyl group having 1 to 10 carbon atoms, an aromatic cyclic acyl group and a carbamido group;

R₂any one selected from hydrogen, a straight chain alkyl group having 1 to 10 carbon atoms, or an alkyl group having a branched chain and having 1 to 10 carbon atoms in the main chain;

LG is selected from any one of chlorine, bromine, iodine, p-toluenesulfonyl, p-nitrobenzenesulfonyl chloride or methanesulfonyl, preferably p-nitrobenzenesulfonyl.

6. The method of claim 5, wherein: it includes:

(1) synthesizing a compound shown in a formula VI with a compound shown in a formula I and ethyl glyoxylate under the action of a catalyst;

(2) carrying out chiral reduction reaction on the compound shown in the formula VI to obtain a compound shown in a formula VII;

(3) carrying out substitution reaction on a compound shown as a formula VII and a sulfonyl chloride compound to obtain a compound shown as a formula VIII;

(4) the compound shown in the formula VIII is subjected to deprotection reaction to obtain a compound shown in the formula XIII;

(5) carrying out bimolecular nucleophilic substitution reaction on a compound shown as a formula XIII and a compound shown as a formula II to obtain a compound shown as a formula XIV;

(6) a compound of formula XIV and said compound S-R₁Or said compound S-R₁Salt passage through the bilayerNucleophilic substitution reaction to obtain the compound shown in the formula AQ-10,

each compound has the following structural formula:

wherein the content of the first and second substances,

p is selected from any one of Boc, Cbz, Fmoc, Bn, PMB or trityl and derivatives thereof, preferably Boc;

the sulfonyl chloride compound is selected from one or more of methylsulfonyl chloride, p-toluenesulfonyl chloride and p-nitrobenzenesulfonyl chloride;

the compound S-R₁The salt is preferably sodium methyl mercaptide.

7. The method of claim 6, wherein: wherein the catalyst of step (1) is selected from morpholine and/or piperidine; preferably, the amount of material of the catalyst is 0.1 to 2 times, more preferably 0.5 to 1.5 times the amount of material of the compound of formula I.

8. The method of claim 6, wherein: wherein the reducing agent for the reduction reaction in step (2) is selected from lithium tri-tert-butoxyaluminum hydride.

9. The method of claim 6, wherein: wherein the deprotection reaction in the step (4) is carried out in a solvent, and the solvent is one or more selected from trifluoroacetic acid, dichloromethane, saturated HCl solution of ethyl acetate and HCl solution of tetrahydrofuran, and is preferably mixed solution of trifluoroacetic acid and dichloromethane.

10. Use of the clopidogrel active metabolite derivative, the prodrug or the pharmaceutically acceptable salt thereof of claims 1 to 4 and/or the clopidogrel active metabolite derivative prepared by the preparation method of claims 5 to 9 for an anti-platelet aggregation drug.

Technical Field

The invention relates to the field of medicinal chemistry, in particular to the technical field of clopidogrel active metabolite derivatives and prodrugs thereof.

Background

Clopidogrel

The non-reversible P2Y12 receptor antagonist is an anti-platelet aggregation prodrug which is most widely applied clinically at present, is mainly applied to various cardiovascular diseases such as atherosclerosis, polar coronary syndrome, thrombotic complications and the like, and has better medicinal activity and less toxic and side effects. But the oral administration bioavailability is low, the effect is slow, the inhibition to blood platelet is delayed, and the clopidogrel resistance phenomenon exists.

Metabolism research finds that the side effects of the drugs generated by clopidogrel are related to the metabolism problem and individual difference, after the existing clopidogrel drugs enter a human body, 85% of the drugs are hydrolyzed by esterase CES1 to form carboxylic acid derivatives without drug activity, and the rest 15% of the drugs are formed by two-step metabolism

Four different configurations of active metabolites, some of the prior studies found that only the fourth configuration had pharmacological activity,

the two-step metabolism process comprises the steps of forming 2-clopidogrel oxide through the oxidation of enzymes such as CYP1A2, CYP2B6 and CYP2C19, and forming active metabolites through the hydrolysis of enzymes such as CYP2C19, CYP2C9, CYP3A4, CYP3A5, PON1 and CYP2B 6. In the second step of metabolism, due to the action of hydrolytic ring opening, metabolites generate three stereochemical sites C7, C3-C16 and C4, and metabolic studies show that the pharmacological activity expression of anti-platelet aggregation of clopidogrel depends on the three sites, wherein the S configuration at the C7 position and the Z configuration of a double bond at the C3-C16 positions are considered to play key roles in exerting the pharmaceutical activity, and the sulfhydryl at the C4 position can be covalently bound with the naked cysteine residue of a P2Y12 receptor of adenosine diphosphate ADP through a disulfide bond, so that the fibrinogen binding site of the platelet protein receptor connected with the receptor is not exposed. Among the above chemical sites, when the double bond at the C3-C16 position is in the E configuration, the resulting metabolite does not have any activity, which is an important cause of low bioavailability of clopidogrel and delay of inhibition.

Disclosure of Invention

The invention aims to provide a series of clopidogrel derivatives based on metabolism, which have high activity, high bioavailability and quick response and can flexibly replace a substituent according to requirements.

The invention also aims to provide a prodrug of the derivative, which can effectively avoid hydrolysis by esterase in vivo, greatly reduces the generation of inactive carboxylic acid metabolites and has small toxic and side effects.

The invention also aims to provide a synthesis method of the derivative and/or the prodrug thereof, which has high stereoselectivity, high yield of a target product and high universality.

The invention firstly provides the following technical scheme:

a clopidogrel active metabolite derivative, a prodrug or a pharmaceutically acceptable salt thereof, wherein the derivative has a structural formula as shown in formula (a):

wherein R is₁Selected from the group consisting of C1-10 linear alkyl, C1-10 branched alkyl, C1-10 alkanoyl, and aromaticAny of cyclic acyl or ureido;

R₂any one selected from hydrogen, a straight chain alkyl group having 1 to 10 carbon atoms, and an alkyl group having a main chain with 1 to 10 carbon atoms and containing a branched chain.

The inventor unexpectedly discovers that compared with the existing clopidogrel active metabolite, the derivative of the invention is deuterated methyl ester at the C7 position, after entering a human body, esterase hydrolysis is effectively avoided, the generation of inactive carboxylic acid derivatives is remarkably reduced, and the bioavailability is high; meanwhile, the composition has the advantages of quick response, high activity and no clopidogrel resistance phenomenon, and remarkably slows down the platelet inhibition delay phenomenon.

According to some embodiments of the present invention, the derivative is (Z) -2- ((S) -1- ((S) -1- (2-chlorophenyl) -2-methoxy-D3-2-oxoethyl) -4- (methylthio) piperidin-3-ylidene) acetic acid represented by the structural formula AQ-10:

the invention further provides a metabolism-based clopidogrel derivative prodrug containing the structure (A), which is one or more of the following compounds:

methyl-D3 (S) -2- ((S, Z) -4-acetylsulfanyl-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

methyl-D3 (S) -2- ((S, Z) -4-isothioureido-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

(Z) -2- ((S) -1- (2-chlorophenyl) -2-methoxy-D3-2-oxoethyl) -4- (methylthio) piperidin-3-ylidene) acetic acid:

methyl-D3 (S) -2- ((S, Z) -4-benzoylsulfanyl-3- (2-ethoxy-2-oxyethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

methyl-D3 (S) -2- ((S, Z) -4-phenylacetylthio-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

methyl-D3 (S) -2- ((S, Z) -4-isopropenylthio-3- (2-ethoxy-2-oxyethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

methyl-D3 (S) -2- ((S, Z) -4-propionylthio-3- (2-ethoxy-2-oxoethyl) piperidin-1-yl) -2- (2-chlorophenyl) acetate:

according to a specific embodiment of the present invention, the prodrug has a structural formula shown in formula (B):

the prodrug forms dimethyl sulfide at the C4 position, so that the toxic and side effects of the prodrug are remarkably reduced while the high-efficiency drug effect of the derivative is maintained, and the structural stability of the derivative is improved.

The invention further provides a preparation method of the clopidogrel active metabolite derivative, which comprises the following steps:

compounds of formula XIV and S-R₁Or the compound S-R₁The salt is obtained by bimolecular nucleophilic substitution reaction (SN2),

wherein R is₁Any one selected from a straight chain alkyl group having 1 to 10 carbon atoms, an alkyl group having a main chain having 1 to 10 carbon atoms and containing a branched chain, a hydrocarbon acyl group having 1 to 10 carbon atoms, an aromatic cyclic acyl group and a carbamido group;

R₂any one selected from hydrogen, a straight chain alkyl group having 1 to 10 carbon atoms, or an alkyl group having a branched chain and having 1 to 10 carbon atoms in the main chain;

LG is selected from any one of chlorine, bromine, iodine, Tos (p-toluenesulfonyl), Ms (methanesulfonyl) or Ns (p-nitrobenzenesulfonyl).

Compounds S-R₁Or a salt thereof is a nucleophilic compound.

According to a specific embodiment of the present invention, the preparation method comprises:

(1) synthesizing a compound shown in a formula VI with a compound shown in a formula I and ethyl glyoxylate under the action of a catalyst;

(2) carrying out chiral reduction reaction on the compound shown in the formula VI to obtain a compound shown in a formula VII;

(3) carrying out substitution reaction on a compound shown as a formula VII and a sulfonyl chloride compound to obtain a compound shown as a formula VIII;

(4) the compound shown in the formula VIII is subjected to deprotection reaction to obtain a compound shown in the formula XIII;

(5) carrying out SN2 substitution reaction on a compound shown as a formula XIII and a compound shown as a formula II to obtain a compound shown as a formula XIV;

(6) compounds of formula XIV and S-R₁Or salts thereof, obtaining the compound (Z) -2- ((S) -1- ((S) -1- (2-chlorphenyl) -2-methoxyl-D3-2-oxyethyl) -4- (methylthio) piperidine-3-methylene) acetic acid shown as the formula AQ-10 by SN2 substitution reaction,

wherein, the structural formula of each compound is as follows:

wherein P is selected from Boc (tert-butyloxycarbonyl), Cbz (benzyloxycarbonyl), Fmoc (fluorenyl-methoxycarbonyl), Bn (benzyl), PMB (P-methoxyphenyl), trityl and derivatives thereof, preferably Boc.

The synthetic route of the preparation method is shown as the following reaction formula:

in the preparation method, the compound S-R in the step (6)₁Or its salt is preferably Na-SCH₃(sodium methyl mercaptide).

The strong basicity of sodium methyl mercaptide can make R₂The ester group is naturally hydrolyzed, so that after the compound shown as the formula AQ is transiently synthesized, the clopidogrel active metabolite derivative shown as the formula AQ-10 can be directly obtained.

According to some embodiments of the invention, the catalyst of step (1) is selected from morpholine and/or piperidine.

The inventors surprisingly found that when morpholine ring and/or piperidine is/are used as a catalyst in the step (1), a compound with double bond Z configuration at C3-C16 position can be synthesized through an enamine intermediate in a reverse steric hindrance specificity manner, high selectivity of a target product is realized, and ee value of the compound can reach more than 95%.

According to some embodiments of the invention, the amount of material of the catalyst is 0.1 to 2 times, preferably 0.5 to 1.5 times the amount of material of the compound of formula I.

According to some embodiments of the invention, the step (2) reduction reaction uses a reducing agent lithium aluminum tri-tert-butoxyhydride.

According to some embodiments of the invention, the VIII compound in step (3) is a sulfonyl chloride compound.

According to some embodiments of the invention, the sulfonyl chloride compound is selected from one or more of methylsulfonyl chloride, p-toluenesulfonyl chloride and p-nitrobenzenesulfonyl chloride.

According to some embodiments of the invention, the deprotection reaction of step (4) is carried out in a solvent selected from one or more of trifluoroacetic acid, dichloromethane, saturated HCl solution of ethyl acetate and HCl solution of tetrahydrofuran.

The inventors have surprisingly found that in the above selection, the use of a mixed solution of trifluoroacetic acid and dichloromethane is more effective than a saturated HCl solution in ethyl acetate and/or a tetrahydrofuran HCl solution.

According to some embodiments of the invention, the leaving group LG of the compound of formula II in step (5) is preferably p-nitrobenzenesulfonyl.

According to further research on the synthesis method of the clopidogrel derivative, the inventor designs a drug intermediate XIV in the reaction, and unexpectedly finds that various derivatives can be effectively synthesized through the strong leaving ability of LG therein, thereby greatly increasing the universality of the synthesis route.

The invention further provides application of the clopidogrel active metabolite derivative, the prodrug or the medicinal salt thereof and/or the clopidogrel active metabolite derivative or the prodrug thereof prepared by the preparation method in an anti-platelet aggregation medicament.

The invention provides a novel clopidogrel active metabolite derivative or a prodrug and a medicinal salt thereof, wherein the active metabolite has high activity, quick response and high bioavailability on inhibiting platelet aggregation, and can avoid the inactive hydrolysis of esterase, thereby remarkably reducing the generation of inactive carboxylic acid metabolites; the prodrug of the invention has stable structure and small toxic and side effects, and simultaneously maintains the high-efficiency drug property of the derivative; the preparation method can selectively synthesize the compound with the double bond Z configuration at the C3-C16 position and the chiral S configuration at the C4 position, and has the advantages of high selectivity, less by-products and high yield.

Drawings

FIG. 1 is a NMR spectrum of a compound VI of example 3 of the present invention;

FIG. 2 is a NMR spectrum of Compound VI of example 3 of the present invention;

FIG. 3 is a schematic diagram of the peak at the C1 position in the carbon spectrum of Compound VI synthesized by the prior art according to example 3 of the present invention;

FIG. 4 is a schematic diagram of the peak at the C1 position in the carbon spectrum of compound VI of example 3 according to the invention;

FIG. 5 is a NMR spectrum of Compound VII shown in example 4 of the present invention;

FIG. 6 is a NMR carbon spectrum of Compound VII shown in example 4 of the present invention;

FIG. 7 is a NMR spectrum of a compound VIII described in example 5 of the present invention;

FIG. 8 is a NMR spectrum of a compound VIII shown in example 5 of the present invention;

FIG. 9 is a NMR spectrum of Compound XIV described in example 7 of the present invention;

FIG. 10 is a NMR carbon spectrum of Compound XIV shown in example 7 of the present invention;

FIG. 11 is a NMR spectrum of Compound AQ-10 according to example 8 of the present invention;

FIG. 12 is a NMR spectrum of AQ-10, a compound shown in example 8 of the present invention;

the drawings are all transversely arranged in the drawings of the specification.

Detailed Description

The present invention is described in detail below with reference to the following embodiments and the attached drawings, but it should be understood that the embodiments and the attached drawings are only used for the illustrative description of the present invention and do not limit the protection scope of the present invention in any way. All reasonable variations and combinations that fall within the spirit of the invention are intended to be within the scope of the invention.

The following examples select the following synthetic routes for the synthesis of clopidogrel active metabolite derivatives:

and selecting the following specific steps:

(1) generating a compound shown as a formula VI by using the compound shown as the formula I and ethyl glyoxylate, wherein the protecting group is Boc;

(2) generating a compound shown in the formula VII from a compound shown in the formula VI through a chiral reduction reaction, wherein the reducing agent is selected to be tri-tert-butoxy lithium aluminum hydride;

(3) substituting a compound of the formula VII by a sulfonyl chloride compound to generate a compound of the formula VIII, wherein the sulfonyl chloride compound is methyl sulfonyl chloride (Ms-Cl) or p-toluene sulfonyl chloride (Tos-Cl) or p-nitrobenzene sulfonyl chloride (Ns-Cl);

(4) carrying out deprotection reaction on the compound shown in the formula VIII to generate a compound XIII;

(5) performing SN2 substitution reaction on a compound of a formula XIII and a compound of a formula II to generate a compound XIV;

(6) the clopidogrel active metabolite derivative AQ-10 is obtained by the reaction of a compound shown in the formula XIV and sodium methyl mercaptide through SN 2.