CN113185483B

CN113185483B - Osthole derivatives and preparation method thereof

Info

Publication number: CN113185483B
Application number: CN202110464698.2A
Authority: CN
Inventors: 孙明娜; 郑雪花; 张建业; 冯晓
Original assignee: Guangzhou Medical University
Current assignee: Guangzhou Medical University
Priority date: 2021-04-28
Filing date: 2021-04-28
Publication date: 2022-10-11
Anticipated expiration: 2041-04-28
Also published as: CN113185483A

Abstract

The invention belongs to the technical field of organic chemistry, and discloses osthole derivatives, a preparation method thereof and application thereof in inhibiting activity of aldehyde ketone reductase 1C 3. The osthole derivative and the pharmaceutically acceptable salt thereof have the structural formula shown as the formula (I) or the formula (II):

the osthole derivative can selectively inhibit the activity of aldehyde ketone reductase 1C3, has obvious inhibition effect, and can be applied to inhibiting aldehyde ketone reductase 1C 3; meanwhile, the final step of androgen generation in tumor cells can be inhibited by inhibiting the activity of aldehyde ketone reductase 1C3, so that the effect of preventing and treating prostatic cancer is achieved, and therefore, the osthole derivative can also be applied to the preparation of medicaments for preventing and treating prostatic cancer diseases.

Description

Osthole derivatives and preparation method thereof

Technical Field

The invention belongs to the technical field of organic chemistry, and particularly relates to osthole derivatives, a preparation method thereof and application thereof in inhibiting activity of aldehyde ketone reductase 1C 3.

Background

Cancer (Cancer), also known as Malignant neoplasms (Malignant neoplasms), is a common and frequently occurring disease that severely threatens human health. Among the male malignancies, the incidence of prostate cancer is the first (28%) and the mortality is 10%, second only to lung cancer. Data show that most prostate cancer patients have metastases already at the time of initial diagnosis. Patients with distant metastases had a 5-year relative survival rate that decreased from 80% to 30% of non-metastatic patients, with half the progression-free survival time of non-metastatic patients. Metastatic hormone sensitive prostate cancer (mHSPC) patients will become metastatic castration resistant prostate cancer (mCRPC) after progressing, and the prognosis is generally poor once patients enter the mCRPC stage.

At this stage, there are many treatments for prostate cancer, including follow-up observation, transurethral prostatectomy (TURP), radical Prostatectomy (RP), radiation therapy, endocrine therapy, comprehensive therapy, and the like. Since abnormalities in Androgen and Androgen receptor pathways contribute to the development and progression of prostate cancer, inhibition of Androgen biosynthesis and blockade of Androgen receptor signaling pathways are the traditional treatment for prostate cancer, namely Androgen Deprivation Therapy (ADT). Initially, ADT is effective for advanced or metastatic prostate cancer, but after a period of treatment, most patients develop fatal castration-resistant prostate cancer (CRPC). The existing medicines for treating CRPC comprise abiraterone and enzalutamide, however, the abiraterone can cause adverse reaction of hypertension, the abiraterone needs to be combined with prednisone, and the two medicines can observe drug resistance in the treatment process. Therefore, the development of CRPC treatment drugs acting on new targets is of great significance.

Aldehyde ketone reductase 1C3 (Aldehyde ketone reductase family 3, AKR1C3) is one of the members of Aldehyde ketone reductase superfamily, also known as type 517-hydroxysteroid dehydrogenase (type 517-hydroxysteroid dehydrogenase,17 beta-HSD 5), catalyzes Nicotinamide Adenine Dinucleotide Phosphate (NADPH) -dependent reduction reaction, and plays an important role in the androgen biosynthesis pathway. Dehydroepiandrosterone (DHEA) can be converted to androstenediol (andrestenediol), androstenedione (AD) to testosterone (testosterone, T), 5a androstenedione (5 α -androsterone) to 5a dihydrotestosterone (5 α -dihydrotestosterone, DHT), and androsterone (androsterone) to androstenediol-3 α (androstanediol-3 α). AKR1C3 also has prostaglandin PGF synthase activity and can promote the growth of tumor cells. Research shows that AKR1C3 is over-expressed in CRPC and plays a certain role in the process of generating drug resistance of cancer cells to enzalutamide and selectively and synergistically activating androgen receptors. Thus, AKR1C3 is considered to be a novel therapeutic target for the treatment of CRPC by inhibiting the final step of androgen production in tumor cells.

In recent years, many reports on AKR1C3 inhibitors have been reported, and the structures of the compounds are diverse, mainly focusing on the following classes of drugs and analogues thereof: steroids (Medroxyprogesterone acetate), flavonoids (2' -hydroxyflavanone), jasmonic acid derivatives (Jasmonic acid), NSAIDs and their analogues (indomethacin analogues, 2-arylpropionic acids (S-Naproxen), benzoic acid derivatives (Flufenamic acid)), cinnamic acid derivatives (Baccharin) and others (SN 33638, ASP9521, GTX-560, morpholine urea derivatives, hydroxytriazole derivatives, benzisoxazole derivatives, berberine, pyrazolopyran derivatives, sulfonylurea drugs (Glimepiride)). However, there is no AKR1C3 inhibitor drug on the market, and ASP9521 for treating CRPC has been clinically tested, but it has been terminated because its clinical efficacy is not obvious. Therefore, the research on the novel potent Gao Zexing AKR1C3 inhibitor is of great significance.

Osthole (osthol) with chemical name of 7-methoxy-8-isopentenyl coumarin, also known as methoxy parsley phenol, parsley methyl ether or oxine, is a linear furocoumarin compound extracted from various Chinese herbal medicines such as cnidium monnieri, radix angelicae pubescentis and the like of herbal plants of Umbelliferae, and is called Osthole because of the most abundant content in dried fruits of cnidium monnieri. Pharmacological research proves that osthole has various biological activities, such as antianaphylaxis, myocardial cell protection, immunity enhancement, osteoporosis resistance and the like. However, no report has been made on AKR1C3 inhibitors of osthole compounds.

Disclosure of Invention

In order to overcome the defects and shortcomings of the prior art, the invention mainly aims to provide an osthole derivative.

The invention also aims to provide a preparation method of the osthole derivative.

The invention further aims to provide the application of the osthole derivative in inhibiting aldehyde ketone reductase 1C 3. The osthole derivative can selectively inhibit the activity of aldehyde ketone reductase 1C3, and has obvious inhibition effect.

The invention also aims to provide application of the osthole derivative in preparing medicaments for preventing and treating prostate cancer diseases. The aldehyde ketone reductase 1C3 plays an important role in an androgen biosynthesis pathway, is over-expressed in CRPC, and plays a role in the processes of drug resistance of cancer cells to enzalutamide and selective synergistic activation of androgen receptors.

The purpose of the invention is realized by the following scheme:

an osthole derivative and pharmaceutically acceptable salts thereof have a structural formula shown as a formula (I) or a formula (II):

wherein the content of the first and second substances,

R ₃ is H or a straight or branched alkyl group containing 1 to 6 carbon atoms;

x is carbonyl or methylene;

y is O or N;

z is heterocyclic radical or alkyl containing one or more of N, O and S;

n is a natural number of 2-4;

R ₁ 、R ₂ the same or different are respectively H, straight chain, branched chain or cyclic alkyl containing 1-6 carbon atoms, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclic radical or heterocyclic aryl containing one or more of N, O and S; or R ₁ 、R ₂ The heterocyclic group or heterocyclic aromatic ring which is substituted or unsubstituted and contains one or more than one N, O or S forms a ring with YAnd (4) a base.

One or more hydrogen atoms in the linear, branched or cyclic alkyl, aryl, heterocyclic or heterocyclic aryl group containing 1 to 6 carbon atoms may be substituted by fluorine atom, oxygen atom, alkenyl group, alkynyl group, aryl group, hydroxyl group, amino group, carbonyl group, carboxyl group, ester group, cyano group, methyl group, ethyl group, methoxy group, nitro group.

When Y is O, R ₁ 、R ₂ Is a substituent.

Further, the osthole derivatives and pharmaceutically acceptable salts thereof have a structure shown as a formula (I) or a formula (II), wherein R is ₁ 、R ₂ The same or different are respectively H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclic group or heterocyclic aryl containing one or more than one of N, O and S; or R ₁ 、R ₂ And Y is a substituted or unsubstituted heterocyclic group or heterocyclic aryl group containing one or more than one of N, O or S.

One or more hydrogen atoms in the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, aryl, heterocyclic group or heterocyclic aryl group may be substituted by a fluorine atom, an oxygen atom, an alkenyl group, an alkynyl group, an aryl group, a hydroxyl group, an amino group, a carbonyl group, a carboxyl group, an ester group, a cyano group, a methyl group, an ethyl group, a methoxy group, a nitro group.

Further, the osthole derivatives and pharmaceutically acceptable salts thereof have a structure shown in a formula (I) or a formula (II), wherein,

when Y is O, R ₁ 、R ₂ Is a substituent selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, substituted or unsubstituted aryl, and benzotriazolyl;

further, one or more hydrogen atoms of the methyl group, the ethyl group, the n-propyl group, the isopropyl group, the n-butyl group, the isobutyl group, the tert-butyl group and the aryl group may be substituted by a fluorine atom, an oxygen atom, an alkenyl group, an alkynyl group, an aryl group, a hydroxyl group, an amino group, a carbonyl group, a carboxyl group, an ester group, a cyano group, a methyl group, an ethyl group, a methoxy group and a nitro group.

When Y is N, R ₁ 、R ₂ The same or different are respectively H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and substituted or unsubstituted aryl.

Or R ₁ 、R ₂ And Y forms a substituted or unsubstituted piperazinyl group, a substituted or unsubstituted pyrrolyl group, and a substituted or unsubstituted piperidyl group.

Still further, one or more hydrogen atoms of the piperazinyl, pyrrolyl and piperidinyl groups may be substituted with a fluorine atom, an oxygen atom, an alkenyl group, an alkynyl group, an aryl group, a hydroxyl group, an amino group, a carbonyl group, a carboxyl group, an ester group, a cyano group, a methyl group, an ethyl group, a methoxy group, or a nitro group.

Furthermore, the osthole derivatives and pharmaceutically acceptable salts thereof have the following structures:

the osthole derivative and the pharmaceutically acceptable salt thereof provided by the invention have the activity of inhibiting aldehyde ketone reductase 1C3, and further inhibit the proliferation of prostate cancer cells.

The invention also provides a preparation method of the osthole derivative, which is shown in the following reaction formula:

reaction formula 1:

reaction formula 2:

reaction formula 3:

reaction formula 4:

reaction formula 5:

wherein the content of the first and second substances,

in the reaction formula 1, osthole is firstly oxidized to obtain a compound 1, and the compound 1 reacts with N-alkylamine to obtain a compound 2; wherein, the oxidant used for oxidation can be conventional oxidants such as selenium dioxide; the reaction of the amine may be carried out under the action of a reducing agent such as sodium cyanoborocyanide.

In the reaction formula 2, the compound 1 is oxidized to obtain a compound 3; reacting the compound 3 with N, N-dialkyl amine to obtain a compound 4 or 4'; wherein, the oxidant used for oxidation can be sodium chlorite and other oxidants which are used for oxidizing aldehyde into acid, and the oxidation reaction is carried out in the acid environment of sodium dihydrogen phosphate; the reaction of the amine can be carried out under the action of benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

In the reaction formula 3, the compound 3 reacts with alkyl alcohol to obtain a compound 5; the reaction can be carried out under the dehydration action of concentrated sulfuric acid.

In the reaction formula 4, the compound 1 is subjected to reduction reaction to obtain a compound 6; dehydrating the compound 6 and alkyl substituted phenol to obtain a compound 7; wherein, the reducing agent adopted in the reduction reaction can be a conventional reduction system such as sodium cyanoborohydride/glacial acetic acid and the like; the dehydration reaction can be carried out in a triphenylphosphine/diethyl azodicarboxylate environment.

In scheme 5, compound 4 is demethylated to afford compound 8. The demethylation may be carried out under the action of boron tribromide.

The process in the preparation method process of the invention is the conventional process in the field, and the adjustment of the process parameters does not influence the reaction, which is known to the technicians in the field; therefore, it is obvious to those skilled in the art that the synthetic route can be determined according to the basic knowledge in the field without departing from the concept of the present invention, and the adopted reactants, catalyst, solvent, temperature and other process parameters can be modified, which all fall into the protection scope of the present invention.

The invention also provides application of the osthole derivative in inhibiting the activity of aldehyde ketone reductase 1C 3. The osthole derivative can selectively inhibit the activity of aldehyde ketone reductase 1C3, and has obvious inhibition effect.

The invention also provides application of the osthole derivative in preparing a medicament for preventing and treating prostatic cancer diseases. The osthole derivative can obviously inhibit the activity of aldehyde ketone reductase 1C3, and achieves the effect of preventing and treating prostatic cancer through the final step of inhibiting the production of androgen in tumor cells.

Furthermore, the invention also provides a pharmaceutical composition for preventing and treating the prostate cancer diseases, which comprises the osthole derivative and/or the pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers and/or excipients.

The carrier can include, for example, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. The composition may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. The composition may be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation or powder.

The pharmaceutical compositions of the invention may be administered parenterally or intraperitoneally, and solutions or suspensions of the free base or pharmaceutically acceptable salts thereof can be formulated in water, suitably mixed with a surfactant, such as hydroxypropylcellulose. The dispersion may be formulated in glycerol, liquid polyethylene glycols and mixtures thereof in oils, which preparations contain a preservative to inhibit the growth of microorganisms under ordinary conditions of storage and use.

The pharmaceutical compositions of the present invention suitable for injectable use may be in the form of sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of injectable solutions or dispersions. In all cases, the form must be sterile and must be flowable to the extent that it is convenient for injection, and must be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms such as bacteria and molds. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, or liquid polyethylene glycol), and suitable mixtures thereof, and vegetable oils.

The osthole derivative can selectively inhibit the activity of aldehyde ketone reductase 1C3 and the proliferation of prostate cancer 22Rv1 cells.

When administered for the treatment or inhibition of a particular disease or condition, it is understood that the effective dosage may vary depending upon the particular compound employed, the mode of administration, the state and severity of the disease, the condition being treated, and various physical factors relating to the individual being treated. In therapeutic applications, the compounds of the present invention are provided to a patient already suffering from a disease in an amount sufficient to treat or at least partially ameliorate the disease and its complications. The appropriate amount is defined as a "therapeutically effective amount". The dose to be used in the treatment in a particular case must be objectively determined by the treating physician, and the changes involved include the particular condition, weight, age and response behavior of the patient. The required dosage level is provided to a human. The dose may be administered once or in two or more doses. The specified daily dosage may vary with the mode of administration. The dosage can be administered in any manner useful for introducing the active compound into the blood of a recipient, including oral, by implant, parenteral (including intravenous, intraperitoneal, intraarticular, and subcutaneous injections), rectal, intranasal, epidermal, intraocular (eye drops), vaginal, and transdermal administration.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a structural formula of an osthole derivative of the present invention.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. The materials referred to in the following examples are commercially available without specific reference. The method is a conventional method unless otherwise specified. The amounts of the components used are in parts by mole, mol, L.

An implementation method, a osthole derivative and pharmaceutically acceptable salts thereof, the structural formula is shown as formula (I) or formula (II):

wherein the content of the first and second substances,

x is carbonyl or methylene;

y is O or N;

z is heterocyclic radical or alkyl containing one or more of N, O and S;

n is a natural number of 2-4;

R ₁ 、R ₂ identical or different are respectively H, C1-C6 straightA chain, branched or cyclic alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heterocyclic or heterocyclic aryl group containing one or more of N, O and S; or R ₁ 、R ₂ And Y is a substituted or unsubstituted heterocyclic group or heterocyclic aryl group containing one or more than one of N, O or S.

When Y is O, R ₁ 、R ₂ Is a substituent.

One or more hydrogen atoms in the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, aryl, heterocyclic group or heterocyclic aryl group can be substituted by fluorine atom, oxygen atom, alkenyl, alkynyl, aryl, hydroxyl, amino, carbonyl, carboxyl, ester group, cyano, methyl, ethyl, methoxy and nitro.

Further, one or more hydrogen atoms of the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and aryl groups may be substituted by fluorine atom, oxygen atom, alkenyl, alkynyl, aryl, hydroxyl, amino, carbonyl, carboxyl, ester group, cyano, methyl, ethyl, methoxy and nitro.

Or R ₁ 、R ₂ And Y form a ring of substituted or unsubstituted piperazinyl, substituted or unsubstituted pyrrolyl, or substituted or unsubstituted piperidinyl.

in one embodiment, the osthole derivatives and pharmaceutically acceptable salts thereof provided by the present invention have activity of inhibiting aldoketoreductase 1C3, thereby inhibiting prostate cancer cell proliferation.

In another embodiment, the present invention further provides a preparation method of the osthole derivative, which is specifically represented by the following reaction formula:

reaction formula 1:

reaction formula 2:

reaction formula 3:

reaction formula 4:

reaction formula 5:

in the reaction formula 1, osthole is firstly oxidized to obtain a compound 1, and the compound 1 reacts with N-alkylamine to obtain a compound 2; wherein, the oxidant used for oxidation can be conventional oxidants such as selenium dioxide and the like; the reaction of the amine may be carried out under the action of a reducing agent such as sodium cyanoborocyanide. As is known to those skilled in the art, osthole derivatives with different substituents can be prepared by carrying out the reaction with different substituted N-alkylamines without affecting the reaction.

In the reaction formula 2, the compound 1 is oxidized to obtain a compound 3; reacting the compound 3 with N, N-dialkyl amine to obtain a compound 4 or 4'; wherein the oxidant used for the oxidation can be sodium chlorite and other conventional aldehyde-oxidizing acidsThe oxidation reaction is carried out in the acid environment of sodium dihydrogen phosphate; the reaction of the amine can be carried out under the action of benzotriazol-1-yl-oxytripyrrolidinylphosphine hexafluorophosphate (PyBOP). In one embodiment, R ₁ And R ₂ Is methyl; in another embodiment, R ₁ And R ₂ Is ethyl; in yet another embodiment, R ₁ 、R ₂ And N forms a ring which is a five-membered ring, a six-membered ring, etc. As is known to those skilled in the art, osthole derivatives with different substituents can be prepared by carrying out the reaction with different substituted N, N-dialkylamines without affecting the reaction.

In the reaction formula 3, the compound 3 reacts with alkyl alcohol to obtain a compound 5; the reaction can be carried out under the dehydration action of concentrated sulfuric acid. In one embodiment, R ₁ Is methyl; in another embodiment, R ₁ Is ethyl; in yet another embodiment, R ₁ Is isopropyl. As is known to those skilled in the art, the osthole derivatives with different substituents can be prepared by carrying out the reaction with different substituted alkyl alcohols without affecting the reaction.

In the reaction formula 4, the compound 1 is subjected to reduction reaction to obtain a compound 6; dehydrating the compound 6 and alkyl substituted phenol to obtain a compound 7; wherein, the reducing agent adopted in the reduction reaction can be a conventional reduction system such as sodium cyanoborohydride/glacial acetic acid and the like; the dehydration reaction can be carried out in a triphenylphosphine/diethyl azodicarboxylate environment. In one embodiment, R ₁ Is a dimethyl-substituted phenyl group; in another embodiment, R ₁ Is methoxy substituted phenyl. As is known to those skilled in the art, osthole derivatives with different substituents can be prepared by carrying out the reaction with different substituted alkyl substituted phenols without affecting the reaction.

The invention also provides application of the osthole derivative in inhibiting the activity of aldehyde ketone reductase 1C 3. The osthole derivative can selectively inhibit the activity of aldehyde ketone reductase 1C3, and has obvious inhibition effect. FIG. 1 is a structural formula of an osthole derivative of the present invention.

Detailed Description

Example 1: preparation of Compound 2a

Step 1: preparation of Compound 1

Weighing osthole crystals (1 molar part) according to a reaction formula 1, and adding 20 volume parts of absolute ethyl alcohol to fully dissolve the osthole; then adding selenium dioxide (1.2 molar parts) into 20 volume parts of N, N-dimethylformamide, and slowly heating while stirring to fully dissolve the selenium dioxide; then dropwise adding the osthole solution into the selenium dioxide solution under stirring, and heating to 90 ℃ for reflux reaction for 10 hours. After the reaction was completed by TLC detection, it was cooled to room temperature and filtered under vacuum to remove the black solid selenium. Reduced pressure steaming and removingMost of the solvent was washed with 20 parts by volume of distilled water, extracted three times with ethyl acetate (20 parts by volume. Times.3), the organic layers were combined, dried over night with anhydrous sodium sulfate, and filtered to remove sodium sulfate. Finally, purifying by column chromatography (eluent PE: EA = 5:1), concentrating and drying to obtain the red powdery solid compound 1 with the yield of 70%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₅ H ₁₅ O ₄ 259.0965,found259.0964. ¹ H NMR(400MHz,DMSO-d ₆ ):δ9.32(s,1H),7.93(d,J＝9.6Hz,1H),7.57(d,J＝8.7Hz,1H),7.05(d,J＝8.7Hz,1H),6.53(d,J＝9.7Hz,1H),6.24(d,J＝9.5Hz,1H),3.87(s,3H),3.70(d,J＝7.4Hz,2H),1.79(s,3H). ¹³ CNMR(101MHz,DMSO-d ₆ ):δ191.43,159.98,159.81,152.37,150.78,144.56,128.04,112.90,112.74,112.44,108.06,56.42,22.40,8.88.

Step 2: preparation of Compound 2a

According to the reaction formula 1, weighing the compound 1 (1 mol part) and dissolving the compound in 20 volume parts of anhydrous tetrahydrofuran, then sequentially adding 3- (4-aminophenyl) ethyl acrylate (1.2 mol parts) and glacial acetic acid (5 mol parts), fully dissolving, adding a molecular sieve for dehydration, and reacting for 2 hours under stirring at room temperature. Sodium cyanoborohydride (2 molar parts) was then added in portions and the reaction was continued with stirring at room temperature for 24h. After TLC detection reaction is completed, 20 volume parts of distilled water is added, washing is carried out, ethyl acetate is used for extraction for three times (20 volume parts multiplied by 3), organic layers are combined, after drying by anhydrous sodium sulfate, the solvent is removed under reduced pressure, column chromatography purification is carried out (eluent PE: EA = 2:1), and the target product is obtained after concentration. Yellow powdery solid 2a, yield 40%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₆ H ₂₈ NO ₅ 434.1962,found434.1957. ¹ HNMR(400MHz,CDCl ₃ ):δ7.61(d,J＝9.5Hz,1H),7.56(d,J＝15.9Hz,1H),7.30(d,J＝8.6Hz,1H),7.27(d,J＝8.1Hz,2H),6.82(d,J＝8.6Hz,1H),6.51(d,J＝8.5Hz,2H),6.23(dd,J＝9.4,0.8Hz,1H),6.17(dd,J＝15.9,0.5Hz,1H),5.48(t,J＝7.2,1H),4.23(q,J＝7.1Hz,2H),3.86(s,3H),3.65(s,2H),3.58(d,J＝7.3Hz,2H),1.89(s,3H),1.32(t,J＝7.1Hz,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ167.99,161.37,160.27,152.93,150.30,145.26,143.85,132.68,129.79,126.60,123.88,123.37,117.07,113.09,113.04,112.83,112.65,107.39,60.15,56.09,51.48,21.73,14.93,14.49.

Example 2: preparation of Compound 4a

Step 1: preparation of Compound 3

According to the reaction formula 2, weighing the compound 1 (1 molar part), adding 20 volume parts of anhydrous tetrahydrofuran, fully dissolving, sequentially adding 2-methyl-2-butene (10 molar parts), sodium dihydrogen phosphate buffer solution (0.4 molar part, 20 volume parts of water) and sodium chlorite (0.4 molar part), stirring under ice bath, reacting for 5 hours, and detecting the reaction completion by TLC. Evaporating under reduced pressure to remove organic solvent, precipitating to obtain solid, vacuum filtering to remove water, washing with small amount of ethyl acetate, and drying to obtain white powdery solid compound 3 with yield of 70%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₅ H ₁₅ O ₅ 275.0914,found 275.0912. ¹ H NMR(400MHz,DMSO-d ₆ ):δ12.17(s,1H),7.91(d,J＝9.6Hz,1H),7.54(d,J＝8.7Hz,1H),7.02(d,J＝8.7Hz,1H),6.53(t,J＝6.8Hz,1H),6.23(d,J＝9.4Hz,1H),3.87(s,3H),3.53(d,J＝7.5Hz,2H),1.90(s,3H). ¹³ C NMR(101MHz,DMSO-d ₆ ):δ168.84,160.10,159.76,144.58,137.74,128.50,127.81,113.66,112.73,108.04,56.40,22.08,12.30.

Step 2: preparation of Compound 4a

According to the reaction formula 2, weighing the compound 3 (1 mol part), adding 30 volume parts of anhydrous tetrahydrofuran to dissolve, sequentially adding dimethylamine (1.2 mol parts), condensing agent PyBOP (1.2 mol parts) and diisopropylethylamine (2 mol parts), stirring at room temperature, and reacting for 3h. After TLC detection reaction is completed, column chromatography purification is carried out, the solvent is removed by evaporation under reduced pressure, and white powdery solid compound 4a is obtained after drying, wherein the yield is 53%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₇ H ₂₀ NO ₄ 302.1387,found 302.1383. ¹ H NMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.6Hz,1H),7.31(d,J＝8.8Hz,1H),6.82(d,J＝8.8Hz,1H),6.22(d,J＝9.2Hz,1H),5.60(t,J＝7.2Hz,1H),3.90(s,3H),3.63(d,J＝7.2Hz,2H),2.92(s,6H),2.04(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ179.10,166.52,165.63,158.35,149.20,137.81,133.02,132.21,121.25,118.39,118.34,112.74,61.48,43.97,40.09,26.83,19.78.

Example 3: preparation of Compound 4b

According to the method for synthesizing the compound 4a of the reaction formula 2, the target compound 4b is synthesized as a white powdery solid with a yield of 62% by using the compound 3 and diethylamine as raw materials. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₉ H ₂₄ NO ₄ 330.1700,found 330.1693. ¹ H NMR(400MHz,CDCl ₃ ):δ7.61(d,J＝8.8Hz,1H),7.31(d,J＝8.8Hz,1H),6.82(d,J＝8.8Hz,1H),6.22(d,J＝9.4Hz,1H),5.60–5.44(m,1H),3.90(s,3H),3.62(d,J＝7.6Hz,2H),3.30(s,4H),2.05(s,3H),1.36-1.24(m,6H). ¹³ C NMR(101MHz,CDCl ₃ ):δ173.26,160.99,160.14,152.91,143.72,132.90,126.72,125.91,115.96,112.89,112.84,107.24,56.01,48.59,38.91,21.27,14.61,14.19,13.02.

Example 4: preparation of Compound 4c

According to the method for synthesizing the compound 4a of the reaction formula 2, the target compound 4c is synthesized as a white powdery solid with a yield of 63% by using the compound 3 and di-n-propylamine as raw materials. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₁ H ₂₈ NO ₄ 358.2013,found 358.2007. ¹ H NMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.6Hz,1H),7.32(d,J＝8.4Hz,1H),6.84(d,J＝8.4Hz,1H),6.23(d,J＝9.6Hz,1H),5.54(t,J＝7.2Hz,1H),3.92(s,3H),3.63(d,J＝7.2Hz,2H),3.23–3.13(m,4H),2.07(m,3H),1.62–1.40(m,4H),0.85–0.62(m,6H). ¹³ CNMR(101MHz,CDCl ₃ ):δ173.70,160.97,160.13,152.85,143.69,132.87,126.71,125.92,115.94,112.94,112.85,107.24,56.00,50.22,45.35,22.07,20.35,21.28,14.72,11.36,10.38.

Example 5: preparation of Compound 4d

According to the method for synthesizing the compound 4a of the reaction formula 2, the target compound 4d is synthesized as a white powdery solid with a yield of 54% by using the compound 3 and di-n-butylamine as raw materials. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₃ H ₃₂ NO ₄ 386.2326,found 386.2325. ¹ HNMR(400MHz,CDCl ₃ ):δ7.60(d,J＝8.3Hz,1H),7.30(d,J＝8.6Hz,1H),6.81(d,J＝8.6Hz,1H),6.21(d,J＝11.1Hz,1H),5.50(t,J＝7.4Hz,1H),3.89(s,3H),3.60(d,J＝7.4Hz,2H),3.24–3.14(m,4H),2.04(s,3H),1.57–1.14(m,6H),1.06–0.60(m,8H). ¹³ CNMR(101MHz,CDCl ₃ ):δ173.54,160.95,160.15,152.85,143.64,132.91,126.68,125.81,115.97,112.95,112.87,107.23,56.01,56.00,48.26,44.04,30.87,29.73,21.27,20.03,19.62,14.66,13.63.

Example 6: preparation of Compound 4e

According to the method for synthesizing the compound 4a in the reaction formula 2, the compound 3 and N-methylpiperazine are used as raw materials to synthesize the target compound 4e, and white powdery solid is obtained with the yield of 67%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₀ H ₂₅ N ₂ O ₄ 357.1089,found357.1084. ¹ HNMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.5Hz,1H),7.32(d,J＝8.6Hz,1H),6.82(d,J＝8.6Hz,1H),6.22(d,J＝9.4Hz,1H),5.59(td,J＝7.2,1.5Hz,1H),3.90(s,3H),3.62(d,J＝7.3Hz,2H),3.53(brs,4H),2.34(brs,4H),2.28(s,3H),2.03(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):172.19,160.97,160.13,152.89,143.74,131.92,127.66,126.81,115.65,112.88,112.88,107.27,56.01,55.14,54.75,46.88,45.89,41.20,21.35,14.61.

Example 7: preparation of Compound 4f

According to the method for synthesizing the compound 4a in the reaction formula 2, the compound 3 and the pyrrolidine are used as raw materials to synthesize the target compound 4f, white powdery solid is obtained, and the yield is 52%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₉ H ₂₂ NO ₄ 328.1543,found328.1538. ¹ HNMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.4Hz,1H),7.31(d,J＝8.6Hz,1H),6.82(d,J＝8.6Hz,1H),6.22(d,J＝9.4Hz,1H),5.72(td,J＝7.3,1.4Hz,1H),3.90(s,3H),3.63(d,J＝7.3Hz,2H),3.43–3.35(m,4H),2.04(s,3H),1.88–1.71(m,4H). ¹³ CNMR(101MHz,CDCl ₃ ):δ171.84,161.07,160.17,152.88,143.75,133.50,128.28,126.75,115.81,112.91,112.87,107.27,56.02,48.61,45.51,26.09,24.32,21.47,13.93.

Example 8: preparation of Compound 4g

According to the method for synthesizing the compound 4a in the reaction formula 2, the compound 3 and 4-methylpiperidine are used as raw materials to synthesize the target compound 4g, and the yield of white powdery solid is 67%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₁ H ₂₆ NO ₄ 356.1856,found356.1849. ¹ HNMR(400MHz,CDCl ₃ ):δ7.61(d,J＝9.5Hz,1H),7.31(d,J＝8.6Hz,1H),6.82(d,J＝8.6Hz,1H),6.22(d,J＝9.5Hz,1H),5.56(td,J＝7.3,1.4Hz,1H),4.61–3.90(m,2H),3.90(s,3H),3.63(d,J＝7.2Hz,2H),2.86–2.61(m,2H),2.03(s,3H),1.82–1.51(m,3H),1.09–0.95(m,2H),0.91(d,J＝6.4Hz,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ172.19,161.02,160.19,152.91,143.74,132.55,126.74,126.67,115.90,112.91,112.88,107.26,56.04,47.15,41.75,34.38,31.14,29.73,21.68,21.34,14.63.

Example 9: preparation of compound 4'h

According to the method for synthesizing the compound 4a by the reaction formula 2, the compound 3 and piperazine are used as raw materials to synthesize the target compound 4'h, and the yield of white powdery solid is 57%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₃₄ H ₃₅ N ₂ O ₈ 599.2388,found599.2380. ¹ HNMR(400MHz,CDCl ₃ ):δ7.63(d,J＝9.5Hz,2H),7.33(d,J＝8.6Hz,2H),6.83(d,J＝8.6Hz,2H),6.23(d,J＝9.4Hz,2H),5.60(t,J＝7.9Hz,2H),3.90(s,6H),3.63(d,J＝7.3Hz,4H),3.44(brs,2H),3.16–3.06(m,2H),2.03(s,6H),1.83–1.74(m,2H),1.44–1.39(m,2H).

Example 10: preparation of Compound 4i

According to the method for synthesizing the compound 4a in the reaction formula 2, the compound 3 and piperazine are used as raw materials to synthesize the target compound 4i, so that white powdery solid is obtained, and the yield is 28%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₉ H ₂₃ N ₂ O ₄ 343.1652,found343.1646. ¹ HNMR(400MHz,DMSO-d ₆ ):δ7.98(d,J＝9.5Hz,1H),7.60(d,J＝8.7Hz,1H),7.08(d,J＝8.7Hz,1H),6.28(d,J＝9.5Hz,1H),5.47(td,J＝7.0,1.4Hz,1H),3.90(s,3H),3.50(d,J＝7.1Hz,2H),3.47–3.38(m,4H),2.87–2.73(m,4H),1.90(s,3H). ¹³ C NMR(101MHz,DMSO-d ₆ ):δ170.82,160.16,159.75,152.32,144.73,131.48,127.69,127.22,114.43,112.73,112.35,108.09,56.39,44.18,21.00,14.19.

Example 11: preparation of Compound 5a

According to the reaction formula 3, weighing the compound 3 (1 molar part), adding 50 volume parts of anhydrous methanol, dropwise adding a small amount of concentrated sulfuric acid, and carrying out reflux reaction for 12 hours.After the TLC detection reaction was completed, the reaction was stopped, the solvent was distilled off under reduced pressure and recrystallized from ethyl acetate/petroleum ether to give white powdery solid 5a with a yield of 71%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₁₆ H ₁₇ O ₅ 289.1070,found 289.1071. ¹ H NMR(400MHz,CDCl ₃ ):δ7.63(d,J＝9.5Hz,1H),7.34(d,J＝8.6Hz,1H),6.84(d,J＝8.6Hz,1H),6.73(t,J＝7.5Hz,1H),6.24(d,J＝9.5Hz,1H),3.91(s,3H),3.71(d,J＝7.7Hz,2H),3.68(s,3H),2.06(s,3H).

Example 12: preparation of Compound 5b

According to the reaction formula 3, weighing the compound 3 (1 molar part), adding 50 volume parts of absolute ethyl alcohol, dropwise adding a small amount of concentrated sulfuric acid, and carrying out reflux reaction for 12 hours. After the TLC detection reaction was completed, the reaction was stopped, the solvent was distilled off under reduced pressure and recrystallized from ethyl acetate/petroleum ether to obtain white powdery solid 5b with a yield of 65%. HRMS (M/z) < M + H] ⁺ calcd.for C ₁₇ H ₁₉ O ₅ 303.1227,found 303.1226. ¹ H NMR(400MHz,CDCl ₃ ):δ7.63(d,J＝9.5Hz,1H),7.34(d,J＝8.6Hz,1H),6.85(d,J＝8.6Hz,1H),6.71(t,J＝6.9Hz,1H),6.25(d,J＝9.3Hz,1H),4.15(q,J＝7.2Hz,2H),3.92(s,3H),3.72(d,J＝7.4Hz,2H),2.06(s,3H),1.25(t,J＝7.1Hz,3H).

Example 13: preparation of Compound 5c

According to the reaction formula 3, weighing the compound 3 (1 molar part), adding 50 volume parts of isopropanol, dropwise adding a small amount of concentrated sulfuric acid, and reacting at 80 ℃ for 12 hours. After the TLC detection reaction was completed, the reaction was stopped, the solvent was distilled off under reduced pressure and recrystallized from ethyl acetate/petroleum ether to obtain white powdery solid 5c with a yield of 45%. ¹ H NMR(400MHz,CDCl ₃ ):δ7.63(d,J＝9.4Hz,1H),7.34(d,J＝8.5Hz,1H),6.84(d,J＝8.6Hz,1H),6.68(t,J＝7.5Hz,1H),6.24(d,J＝9.4Hz,1H),5.04–4.97(m,1H),3.91(s,3H),3.70(d,J＝7.5Hz,2H),2.04(s,3H),1.22(d,J＝6.2Hz,6H).

Example 14: preparation of Compound 5d

The synthesis of compound 4a according to equation 2, without the addition of amine reactant, gave a white solid 5d with a yield of 70%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₁ H ₁₈ N ₃ O ₅ 392.1246,found392.1231. ¹ HNMR(400MHz,CDCl ₃ ):δ8.03(d,J＝8.4Hz,1H),7.66(d,J＝10.3Hz,1H),7.56–7.47(m,1H),7.46–7.35(m,3H),7.28–7.19(m,1H),6.89(d,J＝8.7Hz,1H),6.32–6.23(m,1H),3.97(s,3H),3.89(d,J＝7.5Hz,2H),2.28(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ163.74,160.77,160.24,152.93,145.70,143.67,143.44,128.70,128.55,127.59,124.68,124.13,120.34,113.41,113.24,113.04,108.47,107.36,56.22,23.04,12.70.

Example 15: preparation of Compound 7a

Step 1: preparation of Compound 6

Weighing compound 1 (1 mol part) according to reaction formula 4, dissolving in 20 volume parts of anhydrous tetrahydrofuran, adding glacial acetic acid (5 mol parts), adding a molecular sieve for dehydration, adding sodium cyanoborohydride (3 mol parts) in batches, adding 1 mol part every 1h, adding 3 times, and finally stirring at room temperature for reaction for 24h. After completion of the reaction was checked by TLC, 20 parts by volume of distilled water was added to the reaction solution, washed, extracted three times with ethyl acetate (20 parts by volume × 3), the organic layers were combined, then dried over anhydrous sodium sulfate overnight, and filtered to remove sodium sulfate. Finally, the resulting mixture was purified by column chromatography (eluent DCM: meOH =200: 1), and the solvent was evaporated under reduced pressure from the collected solution and dried to obtain compound 6 as a yellow solid with a yield of 64%. HRMS (M/z) [ M + Na ]] ⁺ calcd.for C ₁₅ H ₁₆ NaO ₄ 283.0941,found283.0942. ¹ HNMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.5Hz,1H),7.30(d,J＝8.6Hz,1H),6.83(d,J＝8.6Hz,1H),6.23(d,J＝9.4Hz,1H),5.49(t,J＝7.8Hz,1H),3.98(s,2H),3.91(s,3H),3.59(d,J＝7.3Hz,2H),1.88(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ161.31,160.18,152.82,143.77,135.73,126.44,122.42,117.06,112.94,112.93,107.29,68.66,56.02,21.42,13.80.

Step 2: preparation of Compound 7a

According to the reaction formula 4, weighing the compound 6 (1 molar part), adding 20 volume parts of anhydrous tetrahydrofuran to dissolve, sequentially adding 3,5-dimethylphenol (2.5 molar parts) and triphenylphosphine (2.5 molar parts), stirring to fully dissolve, and slowly dropwise adding diethyl azodicarboxylate (2.5 molar parts) in an ice bath. The reaction was stirred at room temperature for 24h. After completion of the reaction by TLC detection, the solvent was evaporated under reduced pressure and purified by column chromatography (eluent PE: EA =10: 1) to obtain compound 7a as a white solid with a yield of 60% after concentration. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₃ H ₂₅ O ₄ 365.1747,found365.1747. ¹ HNMR(400MHz,CDCl ₃ ):δ7.62(d,J＝9.5Hz,1H),7.30(d,J＝8.6Hz,1H),6.83(d,J＝8.6Hz,1H),6.55(s,1H),6.51(s,2H),6.24(d,J＝9.4Hz,1H),5.64(t,J＝8.0Hz,1H),4.33(s,2H),3.90(s,3H),3.64(d,J＝7.3Hz,2H),2.24(s,6H),1.95(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ161.23,160.26,159.00,152.85,143.70,138.96,132.30,126.46,124.96,122.34,116.92,113.01,112.95,112.59,107.29,73.76,55.99,21.58,21.38,14.04.

Example 16: preparation of Compound 7b

Compound 7b was synthesized as a white powdery solid in 52% yield according to the procedure for synthesizing compound 7a of reaction formula 4, starting from compound 6 and 3-methoxyphenol. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₂ H ₂₃ O ₅ 367.1540,found 367.1541. ¹ HNMR(400MHz,CDCl ₃ ):δ7.61(d,J＝9.4Hz,1H),7.30(d,J＝8.6Hz,1H),7.12(t,J＝8.1Hz,1H),6.83(d,J＝8.6Hz,1H),6.49–6.44(m,3H),6.24(d,J＝9.5Hz,1H),5.65(td,J＝7.3,1.1Hz,1H),4.34(s,2H),3.90(s,3H),3.74(s,3H),3.63(d,J＝7.3Hz,2H),1.96(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ161.21,160.65,160.25,160.18,152.84,143.70,132.08,129.66,126.49,125.35,116.80,113.00,112.94,107.28,107.00,106.34,101.17,73.95,55.99,55.15,21.58,14.01.

Example 17: preparation of Compound 7c

Compound 7c was synthesized as a white powdery solid in 56% yield according to the procedure for synthesizing compound 7a of reaction formula 4, starting from compound 6 and sesamol. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₂ H ₂₁ O ₆ 381.1331,found 381.1336. ¹ HNMR(400MHz,CDCl ₃ ):δ7.61(d,J＝9.4Hz,1H),7.30(d,J＝8.6Hz,1H),6.83(d,J＝8.6Hz,1H),6.64(d,J＝8.5Hz,1H),6.47(d,J＝0.9Hz,1H),6.29(dd,J＝8.5,2.5Hz,1H),6.24(d,J＝9.5Hz,1H),5.88(s,2H),5.69–5.54(m,1H),4.28(s,2H),3.91(s,3H),3.62(d,J＝7.3Hz,2H),1.94(s,3H). ¹³ CNMR(101MHz,CDCl ₃ ):δ161.21,160.24,154.39,152.84,148.00,143.69,141.48,132.14,126.48,125.30,116.82,113.02,112.95,107.77,107.28,106.29,100.98,98.47,74.99,56.01,21.56,14.00.

Example 18: preparation of Compound 8

According to the reaction formula 5, weighing the compound 4c (1 molar part), adding 20 parts by volume of dichloromethane in an inert atmosphere, adding boron tribromide (5 molar parts) in an ice salt bath, continuing stirring the mixture in the ice salt bath for reaction for 2 hours, and then heating the mixture to room temperature for reaction for 20 hours. Disappearance of the raw material was detected by TLC (developing solvent: petroleum ether/ethyl acetate = 1/1.5) and the reaction was stopped. 20 parts by volume of distilled water was added to the reaction flask,extracting with ethyl acetate for 3 times, mixing several layers, concentrating under reduced pressure, and recrystallizing with petroleum ether/ethyl acetate to obtain light yellow solid compound 8 with yield of 64%. HRMS (M/z) [ M + H ]] ⁺ calcd.for C ₂₀ H ₂₆ NO ₄ 344.1856,found344.1859. ¹ HNMR(400MHz,CDCl ₃ ):δ7.36(d,J＝9.4Hz,1H),6.52(d,J＝8.5Hz,1H),6.30(d,J＝8.5Hz,1H),6.14(d,J＝9.3Hz,1H),5.79(t,J＝7.9Hz,1H),3.49(d,J＝7.9Hz,2H),3.86–3.27(m,4H),2.09(s,3H),1.35–1.18(m,4H),0.94–0.89(m,6H).

Example 19: effect of osthole derivatives on AKR1C3

After incubating the compound of the present invention with a mixed solution containing 0.1M phosphate buffer (pH 6.0), 8. Mu.M 9,10-phenanthrenequinone, and 0.15mM NADPH at 30 ℃ for 10 minutes, recombinant AKR1C3 protein (the recombinant protein is prepared by the method described in chemical-Biological Interactions,2015,240, 310-315) is added and then Flex is used

3 multifunctional microplate reader to detect the effect of compounds on the rate of coenzyme NADPH consumption, each group being measured at least three times. IC for inhibition of AKR1C3 protein activity ₅₀ Values were calculated by concentration testing and non-linear regression.

The data of the test for the inhibitory activity of the compounds of the present invention on AKR1C3 enzyme are shown in table 1.

TABLE 1 inhibition of AKR1C3 by osthole derivatives

As can be seen from table 1, the osthole derivative of the present invention can inhibit the activity of AKR1C3 with a significant inhibitory effect. Wherein, under the same condition, the positive control object Meclofenamic acid (Meclofenamic acid) has the inhibiting activity IC on AKR1C3 enzyme ₅₀ Is 0.52Mu M; IC of Compound 4c, compound 4g of the present invention ₅₀ The value was 0.3 μ M, which is lower than the positive control, meclofenamic acid, and significantly inhibited AKR1C3 activity. The AKR1C3 plays an important role in an androgen biosynthesis pathway, is over-expressed in CRPC, and plays a role in the processes of drug resistance of cancer cells to enzalutamide and selective synergistic activation of androgen receptors.

Example 20: effect of osthole derivatives on AKR1C1

After incubating the compounds 4b and 4C of the present invention with a mixed solution containing 0.1M phosphate buffer (pH 6.0), 8. Mu.M 9,10-phenanthrenequinone, and 0.15mM NADPH at 30 ℃ for 10 minutes, recombinant AKR1C1 protein (the recombinant protein is prepared by the method described in Chemico-Biological Interactions 2015,240, 310-315) is added, followed by Flex

3 multifunctional microplate reader to detect the effect of compounds on the rate of coenzyme NADPH consumption, each group being measured at least three times. IC for inhibiting AKR1C1 protein activity ₅₀ Values were calculated by concentration testing and non-linear regression.

The data for the compounds of the invention for their inhibitory activity against AKR1C1 enzyme are shown in table 2.

TABLE 2 inhibitory Effect of osthole derivatives on AKR1C1

Note: a estimate, from which the inhibition of compounds 4b and 4c was estimated to be 20%, 45%, respectively, at a concentration of 500. Mu.M. b the values are from the reference (J Med chem.2012,55,7746.)

As can be seen from table 2, the osthole derivatives of the present invention can selectively inhibit the activity of AKR1C 3. Wherein, under the condition of 10 mu M, the inhibition rate of the positive control Meclofenamic acid (Meclofenamic acid) to AKR1C1 enzyme is 88.53%; the inhibition rates of the compound 4b and the compound 4C of the invention on AKR1C1 are 4.54 percent and 18.54 percent respectively, which are much lower than the inhibition effect of the positive control meclofenamic acid. Therefore, the osthole derivatives 4b and 4C of the present invention can selectively inhibit AKR1C3 with higher selectivity than the positive control drug.

Example 21: effect of osthole derivatives on prostate cancer 22Rv1 cell proliferation

The proliferation inhibition activity of the osthole derivative of the invention on the in vitro culture of human prostate cancer cell strain 22Rv1 (the cell is purchased from the cell bank of the typical culture occlusion Committee of the Chinese academy of sciences/cell resource center of the Shanghai Life science research institute of the Chinese academy of sciences, catalog number SCSP-5022) is detected by adopting a CCK8 method. Inoculating the tumor cell fluid in logarithmic growth phase into a 96-well plate, wherein the cell density is 1.5 multiplied by 10 ⁴ Per well, 3 multiple wells per drug concentration were set and cultured in RPMI-1640 medium for 24h. Adding 100 μ L of test compound at different concentrations, adding 100 μ L of RPMI-1640 medium to the blank control group, and subjecting the cells to 5% concentration of CO ₂ The cells were cultured in an incubator at 37 ℃ for 72 hours, and the cell viability was measured according to the CCK8 method, and the results are shown in Table 3. As can be seen from Table 3, the osthole derivative of the invention has significant inhibition effect on prostate cancer 22Rv1 cell proliferation.

TABLE 3 proliferation inhibitory Effect of osthole derivatives on prostate cancer 22Rv1 cells

Note: IC of flufenamic acid ₅₀ Values are sourced from the reference eur.j.med.chem.2018,150,930-945.]IC of enzalutamide ₅₀ Values are sourced from reference eur.j.med.chem.2019,171,265-281.]。

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. An osthole derivative and pharmaceutically acceptable salts thereof are characterized in that the structural formula is shown as the formula (I):

wherein the content of the first and second substances,

x is carbonyl or methylene;

when Y is O, R ₁ 、R ₂ Is a substituent selected from ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or benzotriazolyl;

or one or more hydrogen atoms in the ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl are replaced by methyl and ethyl;

when Y is N, R ₁ 、R ₂ Identical or different are each H, a linear, branched or cyclic alkyl radical having from 1 to 6 carbon atoms, or R ₁ 、R ₂ The ring formed with Y is piperazinyl, pyrrolyl or piperidinyl;

or one or more hydrogen atoms in the piperazinyl, the pyrrolyl and the piperidinyl are substituted by methyl and ethyl.

2. Osthole derivatives and pharmaceutically acceptable salts thereof are characterized by having one of the following structures:

3. a process for producing an osthole derivative according to claim 1, which comprises the following reaction scheme 1:

reaction formula 1:

wherein R is ₃ Is methyl; x is methylene; y is N; r ₁ As defined in claim 1, R ₂ Is H.

4. A process for producing an osthole derivative according to claim 1, which comprises the following reaction scheme 2:

reaction formula 2:

wherein R is ₃ Is methyl; x is carbonyl; y is N; r is ₁ 、R ₂ As defined in claim 1.

5. A process for producing an osthole derivative according to claim 2, which comprises the following reaction scheme:

6. a process for producing an osthole derivative according to claim 1, which comprises the following reaction scheme 3:

reaction formula 3:

wherein，R ₃ Is methyl; x is carbonyl; y is O; r ₁ As defined in claim 1.

7. A process for producing an osthole derivative according to claim 2, which comprises the following reaction scheme 4:

reaction formula 4:

wherein the content of the first and second substances,

is composed of

8. A process for producing an osthole derivative according to claim 1, which comprises the following reaction scheme 5:

reaction formula 5:

wherein R is ₃ Is H; x is carbonyl; y is N; r ₁ 、R ₂ As defined in claim 1.

9. Use of an osthole derivative according to any of claims 1-2 and pharmaceutically acceptable salts thereof for the manufacture of a medicament for inhibiting aldoketoreductase 1C3 activity.

10. The use of osthole derivatives and pharmaceutically acceptable salts thereof as claimed in any one of claims 1-2 for the preparation of medicaments for the prevention and treatment of prostate cancer diseases.