A benzothiophenone derivative, a preparation method and application thereof

Abstract

The application discloses a benzothiophenone derivative, a preparation method and application thereof. The benzothiophenone derivative, a tautomer, an optical isomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof, and the chemical structure of the benzothiophenone derivative is shown as formula (I), wherein R is any one of the following groups: a substituted alkane and N-benzyl-2-chloroformamide. The benzothiophenone derivative can regulate a PPARγ target as an anti-type 2 diabetes drug, can reserve insulin sensitivity, can avoid related side effects caused by a PPARγ full agonist, and has a 'highly efficient and safe' hypoglycemic effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a benzothiophenone derivative and a preparation method and application thereof. BACKGROUND

[0002] Peroxisome proliferator-activated receptor gamma (PPARγ) is considered as a key gene in adipocyte biology, which mediates a variety of important physiological functions such as lipogenesis, glucose and lipid metabolism, insulin sensitivity and adipocyte hormone secretion, and plays a core role in the regulation of adipogenesis and insulin sensitivity. Therefore, PPARγ is considered as one of the most promising targets for the treatment of type 2 diabetes. Thiazolidinediones (TZDs) are a class of complete agonists targeting PPARγ, which can improve glucose metabolism, increase insulin sensitivity and significantly reduce blood glucose levels. The listing of TZDs drugs opens a new era of efficient hypoglycemic drugs targeting PPARγ. However, due to its complete agonistic mode of PPARγ, TZDs drugs activate insulin-sensitive genes and also activate the expression of downstream adipocyte differentiation genes, thereby causing obesity, fluid retention, osteoporosis and cardiovascular diseases and other side effects, which adversely affect the health of diabetic patients and greatly limit the application of this class of drugs in today's clinical practice.

[0003] In view of this, more and more recent research has turned to the development of selective PPARγ modulators with partial or no agonistic activity. This class of compounds binds to PPARγ-LBD in a unique way, which can retain anti-diabetic properties while not causing serious side effects. CDK5-mediated phosphorylation of PPARγ-Ser 273 is involved in the pathogenesis of insulin resistance, and phosphorylation at the Ser 273 site leads to abnormal regulation of numerous insulin-sensitive genes downstream of PPARγ, such as reducing the expression of insulin sensitivity-related genes such as Adiponectin and Adipokine, and does not affect PPARγ transcriptional activation. The development of a class of PPARγ partial agonists or non-agonists that inhibit Ser 273 phosphorylation can selectively increase insulin sensitivity while not activating the transcription of side effect-related genes, and the discovery of this mechanism provides a new approach for the development of anti-diabetic drugs targeting PPARγ. SUMMARY

[0004] In order to solve the problems of long development cycle, high cost and low success rate in new drug development, the application provides a benzothiophenone derivative, a preparation method and application thereof, and the inventors develop a new PPAR gamma ligand from a marketed drug library by using high-throughput virtual screening and an old drug new use strategy, and further target design and synthesis of a series of benzothiophenone derivatives by taking the ligand as a mother nucleus structure and comprehensive biological evaluation, so as to obtain a safe candidate compound which can selectively regulate the biological function of PPAR gamma and further play a high-efficiency hypoglycemic effect.

[0005] A first object of the application is to provide a benzothiophenone derivative, a tautomer thereof, an optical isomer thereof, a hydrate thereof, a solvate thereof, a pharmaceutically acceptable salt thereof or a prodrug thereof, the chemical structure of the benzothiophenone derivative being shown as formula (I):

[0006]

[0007] wherein R is any one of the following groups: a substituted alkane, and N-benzyl-2-chloroformamide

[0008] Preferably, the substituted alkane is a halogenated C3-C 10 alkane or C3-C 10 cycloalkane.

[0009] Further preferably, the substituted alkane is specifically a chlorinated or brominated C3-C 10 alkane or C3-C 10 cycloalkane.

[0010] Further preferably, the benzothiophenone derivative is one of the following compounds:

[0011] (4-(hexyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone;

[0012] (5-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(octyloxy)phenyl)methanone;

[0013] (6-N-benzyl-2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-formyl)phenoxy)acetamide;

[0014] (7-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-propyloxyphenyl)methanone;

[0015] (4-butyloxyphenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone;

[0016] (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyl- oxy)phenyl)methanone;

[0017] (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3- yl)methanone;

[0018] (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyl- oxy)phenyl)methanone;

[0019] (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3- yl)methanone;

[0020] (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3- yl)methanone;

[0021] (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyl- oxy)phenyl)methanone;

[0022] (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3- yl)methanone.

[0023] The inventors screened 12 compounds from a marketed drug library (L1000) provided by Shanghai Tauto Biotech Co., Ltd. through high-throughput virtual screening, and determined that the benzothiophenone derivatives YGL-1 to YGL-12 are effective ligands of PPARγ through TR-FRET experiments and SPR experiments. It is further proved that raloxifene has excellent hypoglycemic activity through a series of in vitro experiments such as dual luciferase reporter gene experiments, adipocyte differentiation experiments and RT-qPCR experiments. Subsequently, a series of benzothiophenone derivatives are prepared, and the biological activities thereof are comprehensively determined.

[0024] A second object of the present application is to protect the preparation method of the benzothiophenone derivative, which comprises the following steps: compound 1 (6-methoxy-2-(4-methoxy) benzothiophene) and compound 2 (p-methoxy benzoyl chloride) are subjected to a Friedel-Crafts acylation reaction to obtain an intermediate A, the intermediate A is subjected to a demethylation reaction to obtain an intermediate B, the intermediate B is subjected to an alkylation reaction with a halogenated alkane or N-benzyl-2-chloroformamide to obtain an intermediate C, and the intermediate is subjected to a demethylation reaction to finally obtain a product D, i.e. a benzothiophenone derivative; and the reaction formula is shown as formula (II):

[0025]

[0026] Preferably, the molar ratio of compound 1 to compound 2 is 1:5, the Friedel-Crafts acylation reaction condition is 1-2h at room temperature; the molar ratio of intermediate B to substituted alkane or N-benzyl-2-chloroformamide is 1:2, and the reaction is carried out at room temperature for 12h.

[0027] The preparation method of the above-mentioned benzothiophenone derivative specifically comprises the following steps:

[0028] S1, Friedel-Crafts acylation reaction: a clean and dry 250mL reaction bottle is prepared, 6-methoxy-2-(4-methoxy) benzothiophene (6mmol, 1.0equiv.) is weighed, 100mL of super-dry dichloromethane is added to obtain a dichloromethane solution of 6-methoxy-2-(4-methoxy) benzothiophene with a molar concentration of 0.06mol / L, and then cooled to 0℃, followed by adding p-methoxybenzoyl chloride (9mmol, 1.5equiv.) to the cooled solution. After the reaction is stable, aluminum chloride (9.0mmol, 0.75equiv.) is added to the above mixture, and the reaction is carried out at room temperature for 1-2h. After the reaction is completed, the reaction solution is poured into ice water (150mL) for quenching, and the solution is extracted with dichloromethane, 1M NaOH solution and saturated brine, dried over anhydrous sodium sulfate, and the solvent is removed. Finally, intermediate A is purified by flash silica gel column chromatography, and the eluent is (PE / EA=3:1).

[0029] S2, demethylation reaction: a clean and dry 250mL three-necked flask is prepared, sodium ethanethiol (15mmol, 1.5equiv.) is added after nitrogen replacement, intermediate A (10mmol, 1equiv.) is dissolved in super-dry 50mL DMF to obtain a DMF solution of compound A with a concentration of 0.2mol / L, which is added to the round-bottom flask, and the reaction is carried out at 80℃ for 5h. The reaction is monitored by TLC until the reaction is completed. After the reaction is completed, it is cooled to room temperature, acidified with 10% HCl solution, extracted with diethyl ether, and the water layer is extracted again with diethyl ether. The combined diethyl ether layer is extracted with water and 5% NaOH, and the diethyl ether layer is retained, dried over anhydrous sodium sulfate, and the solvent is removed to obtain intermediate B.

[0030] S3, condensation reaction: take a clean 25 mL round-bottom flask, add intermediate B (0.2 mmol, 1 equiv.), cesium carbonate (0.4 mmol, 2 equiv.), potassium iodide (0.4 mmol, 2 equiv.), RX (0.4 mmol, 2 equiv.) and DMF (3 mL) into the flask, the molar ratio of intermediate B and RX is 1:2, stir at room temperature for 12 h, monitor by TLC, when the reaction is completed, add ice water for quenching, then extract with EA for 2-3 times, combine the organic layers, dry over anhydrous sodium sulfate, remove the solvent. Finally, purify the intermediate C by flash silica gel column chromatography, eluent: (PE / EA = 1:1). RX is halogenated alkane or N-benzyl-2-chloroformamide.

[0031] S4, demethylation reaction: prepare a dry and clean three-necked flask, under N2 atmosphere, dissolve intermediate C (0.2 mmol, 1 equiv.) in DCM (5 mL) to obtain a DCM solution with a molar concentration of 0.04 mol / L and add it into the flask, and place it in a low-temperature tank. When the temperature is lowered to -78℃, add BBr3 (1.2 mmol, 6 equiv.), and when the reaction is stable, slowly warm it to room temperature. Stir the mixture for 0.5-2 h, monitor the reaction by TLC (PE / EA = 1:1), and when the reaction is completed, quench it at 0℃ by adding water, extract the mixture with EA, separate and concentrate the organic layer, and purify to obtain product D.

[0032] A third object of the present application is to protect the use of benzothiophenone derivatives, their tautomers, their optical isomers, their hydrates, their solvates, their pharmaceutically acceptable salts or their prodrugs in the preparation of anti-diabetic drugs. The benzothiophenone derivatives are benzothiophenone derivatives represented by the above formula (I) or 2-(4-hydroxyphenyl)-3-{4-[2-(1-piperidinyl)ethoxy]benzoyl}-6-hydroxybenzo[b]thiophene hydrochloride. The chemical name of raloxifene proposed in the present application is: 2-(4-hydroxyphenyl)-3-{4-[2-(1-piperidinyl)ethoxy]benzoyl}-6-hydroxybenzo[b]thiophene hydrochloride.

[0033] Preferably, the anti-diabetic drug is a PPARγ-targeted anti-type II diabetes drug.

[0034] The benzothiophenone derivative can effectively play a role in reducing blood sugar, regulate the expression of downstream target genes by regulating the PPARγ target, and play a high-efficiency role in reducing blood sugar which can retain insulin-sensitizing effect and avoid related side effects caused by PPARγ full agonists. At present, the thiazolidinedione (TZDs) oral hypoglycemic drugs (such as rosiglitazone-avandia) with PPARγ as a target have a strong effect on improving insulin sensitivity, which brings a major breakthrough for T2DM treatment. TZDs are PPARγ However, the use of TZDs drugs in clinical use is accompanied by the occurrence of side effects such as weight gain, fluid retention, myocardial hypertrophy, liver toxicity and osteoporosis, which seriously hinders the use of TZDs drugs in clinical use. Therefore, it is an urgent need in the medical field to find a kind of oral hypoglycemic drugs which can retain insulin-sensitizing effect and reduce related side effects.

[0035] The benzothiophenone derivative can effectively play a role in reducing blood sugar, regulate the expression of downstream target genes by regulating the PPARγ target, and play a high-efficiency role in reducing blood sugar which can retain insulin-sensitizing effect and avoid related side effects caused by PPARγ full agonists. At present, the thiazolidinedione (TZDs) oral hypoglycemic drugs (such as rosiglitazone-avandia) with PPARγ as a target have a strong effect on improving insulin sensitivity, which brings a major breakthrough for T2DM treatment. TZDs are PPARγ However, the use of TZDs drugs in clinical use is accompanied by the occurrence of side effects such as weight gain, fluid retention, myocardial hypertrophy, liver toxicity and osteoporosis, which seriously hinders the use of TZDs drugs in clinical use. Therefore, it is an urgent need in the medical field to find a kind of oral hypoglycemic drugs which can retain insulin-sensitizing effect and reduce related side effects.

[0036] The fourth object of the present application is to protect an anti-diabetic drug containing an effective amount of the benzothiophenone derivative shown in the above formula (I) or 2-(4-hydroxyphenyl)-3-{4-[2-(1-piperidinyl)ethoxy]benzoyl}-6-hydroxybenzo[b]thiophene hydrochloride (i.e. raloxifene) as an active ingredient and a pharmaceutically acceptable carrier.

[0037] Further preferably, the anti-diabetic drug is an anti-type II diabetes drug with PPARγ as a target.

[0038] Compared with the prior art, the present application has the following advantages: the benzothiophenone derivative can be used as an anti-type II diabetes drug by regulating the PPARγ target, and can play a "highly safe" role in reducing blood sugar which can retain insulin-sensitizing effect and avoid related side effects caused by PPARγ full agonists. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 is the ability of the compound raloxifene in experimental example 1 to activate PPARγ;

[0040] Figure 2 is a statistical graph of the compound raloxifene in experimental example 1 inducing adipocyte differentiation;

[0041] Figure 3 is the expression of related genes in experimental example 1;

[0042] Figure 4 is the PPARγ activation ability of benzothiophenone derivatives 1-12 and raloxifene in Experimental Example 1;

[0043] Figure 5 is the PPARγ binding ability of benzothiophenone derivatives 1-12 and raloxifene in Experimental Example 1;

[0044] Figure 6 is the adipocyte differentiation induced by benzothiophenone derivative YGL-12 in Experimental Example 1;

[0045] Figure 7 is the expression of downstream related genes under the action of benzothiophenone derivative YGL-12 in Experimental Example 1;

[0046] Figure 8 is the fasting body weight, blood glucose and glucose tolerance of mice under the action of benzothiophenone derivative YGL-12 and raloxifene in Experimental Example 1;

[0047] Figure 9 is the organ weight of mice and the general morphology of mice under the action of benzothiophenone derivative YGL-12 and raloxifene in Experimental Example 1;

[0048] In the above figure, YGL-1, YGL-2, YGL-3, YGL-4, YGL-5, YGL-6, YGL-7, YGL-8, YGL-9, YGL-10, YGL-11 and YGL-12 correspond to compounds 1-12 in Examples 1-12 below respectively. DETAILED DESCRIPTION

[0049] The present application will be further explained in conjunction with the following examples, but the examples do not limit the present application in any form.

[0050] Examples

[0051] The preparation method of benzothiophenone derivatives specifically comprises the following steps:

[0052] S1, Friedel-Crafts acylation: prepare a clean and dry 250 mL reaction flask, weigh 6-methoxy-2-(4-methoxy) benzothiophene (6 mmol, 1.0 equiv.), add 100 mL of super dry dichloromethane to obtain a 0.06 mol / L molar concentration of 6-methoxy-2-(4-methoxy) benzothiophene dichloromethane solution, cool to 0°C, then add p-methoxybenzoyl chloride (9 mmol, 1.5 equiv.) to the cooled solution. After the reaction is stable, add aluminum chloride (9.0 mmol, 0.75 equiv.) to the above mixture, react at room temperature for 1-2 h, and after the reaction is complete, pour the reaction solution into ice water (150 mL) for quenching, extract the solution with dichloromethane, 1M NaOH solution and saturated brine, dry over anhydrous sodium sulfate, and remove the solvent. Finally, purify the intermediate A by flash silica gel column chromatography, eluent: (PE / EA = 3:1).

[0053] S2, demethylation reaction: prepare a clean and dry 250 mL three-necked flask, after adding sodium ethanethiol (15 mmol, 1.5 equiv.) and replacing nitrogen, dissolve intermediate A (10 mmol, 1 equiv.) in 50 mL of super dry DMF to obtain a 0.2 mol / L concentration of compound A in DMF solution, add to the round-bottom flask, and react at 80°C for 5 h, monitor the reaction by TLC until the reaction is complete. After the reaction is complete, cool to room temperature, acidify with 10% HCl solution, extract with ether, extract the aqueous layer with ether again, combine the ether layers and extract with water and 5% NaOH, retain the ether layer, dry over anhydrous sodium sulfate, remove the solvent to obtain intermediate B.

[0054] S3, condensation reaction: take a clean 25 mL round-bottom flask, add intermediate B (0.2 mmol, 1 equiv.), cesium carbonate (0.4 mmol, 2 equiv.), potassium iodide (0.4 mmol, 2 equiv.), 1-(2-chloroethyl) piperidine hydrochloride (0.4 mmol, 2 equiv.) and DMF (3 mL) to the flask, the molar ratio of intermediate B and RX is 1:2, stir at room temperature for 12 h, monitor by TLC, after the reaction is complete, add ice water for quenching, then extract with EA for 2-3 times, combine the organic layers, dry over anhydrous sodium sulfate, and remove the solvent. Finally, purify the intermediate C by flash silica gel column chromatography, eluent: (PE / EA = 1:1).

[0055] S4, demethylation reaction: prepare a dry and clean three-necked flask, under N2atmosphere, dissolve intermediate C (0.2 mmol, 1 equiv.) in DCM (5 mL) to obtain a DCM solution with a molar concentration of 0.04 mol / L and add it to the flask, and place it in a low-temperature tank, and wait until it is cooled to -78°C, then add BBr3(1.2 mmol, 6 equiv.), after the reaction is stable, slowly warm it to room temperature, stir the mixture for 0.5-2 h, monitor the reaction by TLC (PE / EA = 1:1), after monitoring the completion of the reaction, quench it with water at 0°C, extract the mixture with EA, separate and concentrate the organic layer, and purify to obtain the product D 2-(4-hydroxyphenyl)-3-{4-[2-(1-piperidinyl)ethoxy]benzoyl}-6-hydroxybenzo[b]thiophene hydrochloride (i.e. Raloxifene).

[0056] Example 1

[0057]

[0058] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 is replaced with 1-bromohexane, and (4-(hexyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 63%).

[0059] 1 H NMR (400 MHz, DMSO-d6): δ 7.65 (d, J = 8.9 Hz, 2H), 7.34 (d, J = 2.2 Hz, 1H), 7.26 (d, J = 8.8 Hz, 1H), 7.18 (d, J = 8.6 Hz, 2H), 6.95-6.78 (m, 3H), 6.68 (d, J = 8.6 Hz, 2H), 3.94 (t, J = 6.5 Hz, 2H), 1.65 (p, J = 6.6 Hz, 2H), 1.47-1.29 (m, 2H), 1.32-1.11 (m, 4H), 0.83 (t, J = 6.8 Hz, 3H).

[0060] Example 2

[0061]

[0062] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 is replaced with 1-bromohexane, and (4-(hexyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 63%).

[0063] 1H NMR (400 MHz, DMSO-d6): δ 9.80 (s, 2H), 7.65 (d, J = 8.9 Hz, 2H), 7.35 (s, 1H), 7.26 (d, J = 8.8 Hz, 1H), 7.18 (d, J = 8.7 Hz, 2H), 6.86 (d, J = 9.0 Hz, 3H), 6.68 (d, J = 8.7 Hz, 2H), 3.97-3.86 (t, J = 6.4 Hz, 2H), 1.72-1.52 (m, 2H), 1.36-1.26 (m, 2H), 1.24-1.14 (m, 8H), 0.81 (t, J = 7.0 Hz, 3H).

[0064] Example 3

[0065]

[0066] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by N-benzyl-2-chloroformamide, N-benzyl-2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)acetamide (yellow solid, yield 68%).

[0067] 1 H NMR (400 MHz, DMSO-d6): δ 9.77 (s, 2H), 8.67 (t, J = 6.1 Hz, 1H), 7.68 (d, J = 8.9 Hz, 2H), 7.34 (d, J = 2.1 Hz, 1H), 7.30-7.16 (m, 8H), 6.96 (d, J = 9.0 Hz, 2H), 6.85 (dd, J = 8.8, 2.3 Hz, 1H), 6.70-6.65 (m, 2H), 4.60 (s, 2H), 4.32 (d, J = 6.1 Hz, 2H).

[0068] Example 4

[0069]

[0070] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by 1-bromopropane, (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-yl)(4-propoxyphenyl)methanone (yellow solid, yield 65%).

[0071] 1H NMR (400MHz, DMSO-d6): δ9.85(d,J=19.4Hz,2H),7.67-7.64(m,2H),7.36(d,J=2.3Hz,1H),7.26(d,J=9.0Hz,1H),7.19- 7.15(m,2H),6.90-6.87(m,2H),6.71-6.67(m,2H),3.92(t,J=6.5Hz,2H),1.68(h,J=7.3Hz,2H),0.92(t,J=7.4Hz,3H).

[0072] Example 5

[0073]

[0074] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by 1-bromobutane, (4-butoxyphenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 56%).

[0075] 1 H NMR (400MHz, DMSO-d6): δ9.75(s,2H),7.65(d,J=8.6Hz,2H),7.34(s,1H),7.25(d,J=8.7Hz,1H),7.17(d,J=8.4Hz,2H),6.87(dd,J= 12.9,7.8Hz,3H),6.68(d,J=8.4Hz,2H),3.96(t,J=6.4Hz,2H),1.65(p,J=6.7Hz,2H),1.38(h,J=7.4Hz,2H),0.89(t,J=7.4Hz,3H).

[0076] Example 6

[0077]

[0078] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced with 1-bromopentane, ((6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyloxy)phenyl)methanone) (yellow solid, yield 50%).

[0079] 1H NMR (400 MHz, DMSO-d6): δ 9.79 (s, 2H), 7.67-7.63 (m, 2H), 7.34 (d, J = 2.6 Hz, 1H), 7.25 (d, J = 8.7 Hz, 1H), 7.19-7.14 (m, 2H), 6.90-6.84 (m, 3H), 6.69-6.66 (m, 2H), 3.95 (t, J = 6.5 Hz, 2H), 1.66 (p, J = 6.6 Hz, 2H), 1.36-1.26 (m, 4H), 0.89-0.82 (m, 3H).

[0080] Example 7

[0081]

[0082] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced with 1-bromoheptane, and (4-(heptyloxy)phenyl)(6-hydroxy-2-(4- hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 47%).

[0083] 1 H NMR (400 MHz, DMSO-d6): δ 9.80 (s, 1H), δ 9.75 (s, 1H), 7.67-7.62 (m, 2H), 7.34 (d, J = 2.3 Hz, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.18-7.15 (m, 2H), 6.91-6.87 (m, 2H), 6.85 (dd, J = 8.8, 2.3 Hz, 1H), 6.69-6.65 (m, 2H), 3.96 (t, J = 8.0 Hz, 2H), 1.67 (p, J = 6.5 Hz, 2H), 1.34 (dt, J = 10.5, 5.4 Hz, 2H), 1.24 (td, J = 7.8, 7.2, 3.7 Hz, 6H), 0.84 (t, J = 4.0 Hz, 3H).

[0084] Example 8

[0085]

[0086] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced with 1-bromononane, and (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(nonyloxy)phenyl)methanone (yellow solid, yield 49%).

[0087] 1H NMR (400 MHz, DMSO-d6): δ 9.79 (s, 2H), δ 9.76 (s, 2H), 7.65 (d, J = 8.9 Hz, 2H), 7.34 (d, J = 2.2 Hz, 1H), 7.24 (d, J = 8.7 Hz, 1H), 7.20-7.14 (m, 2H), 6.88 (d, J = 8.9 Hz, 2H), 6.85 (dd, J = 8.8, 2.3 Hz, 1H), 6.70-6.63 (m, 2H), 3.95 (t, J = 6.5 Hz, 2H), 1.65 (p, J = 6.5 Hz, 2H), 1.39-1.31 (m, 2H), 1.28-1.22 (m, 10H), 0.88-0.78 (m, 3H).

[0088] Example 9

[0089]

[0090] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by 1-bromodecane, and (4-(decyloxy)phenyl)(6-hydroxy-2-(4- hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 36%).

[0091] 1 H NMR (400 MHz, DMSO-d6): δ 9.78 (s, J = 16.5 Hz, 1H), 9.74 (s, J = 16.5 Hz, 1H), 7.68-7.61 (m, 2H), 7.33 (s, 1H), 7.24 (d, J = 8.7 Hz, 1H), 7.20-7.14 (m, 2H), 6.88 (d, J = 8.9 Hz, 2H), 6.85 (dd, J = 8.8, 2.3 Hz, 1H), 6.69-6.65 (m, 2H), 3.95 (t, J = 6.5 Hz, 2H), 1.66 (p, J = 6.6 Hz, 2H), 1.35 (p, J = 8.1 Hz, 2H), 1.30-1.21 (m, 12H), 0.87-0.80 (t, J = 14.9 Hz, 3H).

[0092] Example 10

[0093]

[0094] Referring to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by 3-bromohexane, and (4-(hexan-3-yloxy)phenyl)(6-hydroxy-2-(4- hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 52%).

[0095] 1 H NMR (400 MHz, DMSO-d6): δ 9.79 (s, 1H), δ 9.76 (s, 1H), 7.67-7.63 (m, 2H), 7.34 (dd, J = 2.3, 0.4 Hz, 1H), 7.25-7.22 (m, 1H), 7.19-7.15 (m, 2H), 6.93-6.90 (m, 2H), 6.84 (dd, J = 8.8, 2.3 Hz, 1H), 6.69-6.66 (m, 2H), 3.87 (d, J = 5.8 Hz, 2H), 1.64 (p, J = 6.0 Hz, 1H), 1.40-1.31 (m, 2H), 1.26-1.23 (m, J = 9.1, 5.6 Hz, 4H), 0.85 (t, J = 7.5 Hz, 6H).

[0096] Example 11

[0097]

[0098] With reference to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by 3-bromopropylbenzene, (6-hydroxy-2-(4- hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(3-phenylpropoxy)phenyl)methanone (yellow solid, yield 50%).

[0099] 1 H NMR (400 MHz, DMSO-d6): δ 9.74 (s, 2H), 7.66 (d, J = 8.9 Hz, 2H), 7.34 (d, J = 2.2 Hz, 1H), 7.29-7.23 (m, 3H), 7.21-7.15 (m, 5H), 6.91-6.84 (m, 3H), 6.72-6.66 (m, 2H), 3.97 (t, J = 6.3 Hz, 2H), 2.73-2.66 (m, 2H), 1.98 (dt, J = 13.5, 6.3 Hz, 2H).

[0100] Example 12

[0101]

[0102] With reference to the above preparation method, 1-(2-chloroethyl)piperidine hydrochloride in step S3 was replaced by bromomethylcyclohexane, (4-(cyclohexylmethoxy)phenyl)(6- hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone (yellow solid, yield 42%).

[0103] 1H NMR (400 MHz, DMSO-d6): δ 9.76 (s, 2H), 7.65 (d, J = 8.9 Hz, 2H), 7.34 (s, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.20-7.14 (m, 2H), 6.94-6.80 (m, 3H), 6.73-6.63 (m, 2H), 3.79 (d, J = 6.2 Hz, 2H), 1.81-1.57 (m, 6H), 1.30-1.08 (m, 4H), 0.99 (q, J = 12.9 Hz, 2H).

[0104] Experimental Example 1

[0105] This experimental example is used to illustrate the use of the compounds obtained in Examples 1-12.

[0106] 1. TR-FRET assay to detect the binding ability of compound Raloxifene to PPARγ, the specific experimental steps are as follows: first, prepare the compound stock solution and dilute to the corresponding concentration; prepare the standard Buffer solution, prepare the test compound, and the corresponding concentration reagent.

[0107] (1) In a 384-well plate, sequentially add the test compound (20 μL 2xTest Compound, 10 μL 4xFluormone TM Pan-PPAR Green, 10 μL 4xPPARγ-LBD / Tb-anti-GST Ab);

[0108] (2) Positive control (20 μl 2xControl Competitor), negative control (20 μl 2xTest Compound Solvent), the same as above;

[0109] (3) After addition, wrap the plate with tin foil to avoid light exposure and prevent liquid evaporation, and incubate at room temperature for 4-6 h on a shaker;

[0110] (4) Use a multifunctional enzyme marker to detect the fluorescence absorption signal at 495 nm and 520 nm, respectively;

[0111] (5) Calculate the TR-FRET ratio by the ratio of the absorbance values at 520 nm and 495 nm.

[0112] 2. Dual luciferase activity assay to detect the PPARγ activation ability of high-throughput virtual screening compounds, the specific experimental steps are as follows:

[0113] Cos-7 cells were purchased from ATCC and cultured in DMEM supplemented with 10% FBS at 37°C in a 5% CO2 incubator. After the cells entered the logarithmic growth phase, they were seeded in 24-well plates. When cell confluency reached approximately 70%, they were co-transfected with 50 ng of full-length hPPARγ, 100 ng of PPARγ, and 5 ng of Renilla luciferase plasmid using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. Twenty-four hours later, transfected cells were treated with 10 μM of each compound, with 10 μM rosiglitazone serving as a positive control and DMSO as a negative control. Luciferase activity was measured 24 hours after treatment using the Reporter Luciferase assay kit (Promega) with three independent wells per group.

[0114] Dual-luciferase reporter gene assay

[0115] Prepare cell lysis buffer: Mix Passive Lysis Buffer (PLB) and ddH2O at a volume ratio of 1:4, add to a 24-well plate, and incubate at room temperature for 20 minutes; Prepare Luciferase Assay Reagent II: Mix 1 vial Luciferase Assay Substrate with 10 mL Luciferase Assay Reagent II; Prepare Reagent: 50× Substrate and 50 volume Buffer was mixed evenly (prepared and used immediately); 20 μL of PLB lysis buffer containing cell lysate was added to a white opaque 96-well plate, followed by 100 μL of Luciferase Assay Reagent II. After mixing, firefly luciferase activity was detected using a microplate reader; immediately after the detection was completed, 100 μL Reagent, and immediately detect the Renilla luciferase activity after mixing; calculate the activity ratio of the two tests (first / second) to obtain the transcriptional activation data.

[0116] The experimental results show that ( Figure 1 ), compared with Rosi, Raloxifene only weakly activated PPARγ.

[0117] 3. Oil red O staining to detect the ability of the compound to induce adipocyte differentiation. The specific experimental steps are as follows: 3T3-L1 preadipocytes are purchased from ATCC and cultured in DMEM containing 10% FBS at 37°C in a 5% CO2 incubator. When the cells enter the logarithmic growth phase, they are inoculated in a 6-well plate. After confluence, the cells are cultured for another 2 days, and then the induction solution (10% FBS DMEM containing 0.5 mmol / L IBMX (3-isobutyl-1-methylxanthine), 1 μmol / L DEX (dexamethasone), and 850 nmol / L insulin) is added. After 72 h, the medium is replaced with 10% FBS high-sugar DMEM containing 850 nmol / L insulin, and the medium is replaced every 2 days. 10 μM rosiglitazone is used as a positive control, and DMSO is used as a negative control. The sample groups are 10 μM Raloxifene and YGL-12. Oil red O staining is performed on the 8th day of induction, and the differentiation rate is calculated using Image J software under a microscope.

[0118] The experimental results show that Figure 2 , Table 1), and the results show that the top 10 compounds provided by the present application that bind to PPARγ weakly induce adipocyte differentiation only slightly compared with the positive control drug rosiglitazone (Rosi). Among them, the compound Raloxifene (adipocyte differentiation rate 8.97%) can significantly inhibit the effect of the positive drug rosiglitazone (adipocyte differentiation rate 41.86%) on inducing adipocyte differentiation, indicating that the compounds proposed in the present application can better avoid the side effects caused by the positive rosiglitazone.

[0119] Table 1. Raloxifene-induced adipocyte differentiation rate table

[0120]

[0121]

[0122] 4. Real time PCR to detect the expression of lipogenic genes. The specific steps are as follows: 3T3-L1 preadipocytes are induced and differentiated according to the above-mentioned scheme, total RNA is extracted, and Real time PCR is performed according to the kit (Takara) scheme (Table 2). The method of ΔΔ-Ct is used, β-actin is used as an internal reference, and the relative expression of mRNA is calculated.

[0123] The expression of adipocyte differentiation-related genes is detected by RT-qPCR, and the specific operation is as follows:

[0124] (1) The cells are rinsed with PBS twice, 800 μL / well of Trizol is added, and the cells are detached by blowing with a pipette gun. Then, it is transferred to a 1.5 mL EP tube, and it is left at room temperature for 10 min;

[0125] (2) Add 300 μL of chloroform, cover the EP tube tightly, shake vigorously up and down for 15 seconds to mix thoroughly, let it stand at room temperature for 5 minutes, and then centrifuge at 12000 rpm at 4°C for 15 minutes;

[0126] (3) After centrifugation, the sample tube is separated into three layers. The upper aqueous phase is retained and transferred to a new EP tube;

[0127] (4) Add an equal volume of pre-cooled isopropanol to the sample tube, gently invert to mix, and let stand at room temperature for 10 minutes;

[0128] (5) Centrifuge at 12,000 rpm at 4°C for 10 min. The white precipitate formed at the bottom is RNA. Discard the supernatant and add 1 mL of 75% ethanol (prepared with DEPC water) to wash the RNA precipitate (gently invert to avoid blowing away the precipitate). Centrifuge at 10,000 rpm at 4°C for 5 min.

[0129] (6) Repeat step (5) once, and finally remove 75% ethanol, leaving the RNA precipitate, and air-dry it in a clean bench (30 min); add appropriate amount of DEPC water to dissolve the RNA, and quantify it using Nanodrop 2000.

[0130] Table 2 qPCR primer sequences

[0131]

[0132]

[0133] The experimental results show that ( Figure 3 Compared with the control group, Raloxifene only weakly activated the expression of downstream adipogenesis-related genes and selectively upregulated the expression of the insulin-sensitive gene Adiponectin. While the Rosi group also activated the expression of insulin-sensitive genes, it also significantly activated the expression of genes involved in adipocyte differentiation.

[0134] After confirming that Raloxifene is an effective PPARγ ligand, based on the structure, a series of benzothiophene ketone derivatives were prepared with the benzothiophene ketone structure of raloxifene as the parent core. Their biological activities were tested through a series of in vitro and in vivo experiments. The specific implementation plan is the same as above. The experimental results are as follows: Figure 6 and as shown in Table 3.

[0135] Table 3 Statistics of YGL-12-induced adipocyte differentiation

[0136] DMSO 7.55％ Rosi 41.86％ YGL-12 6.68％ YGL-12 + Rosi 13.58％

[0137] 5. The hypoglycemic effect of the compound raloxifene and YGL-12 is detected in the db / db model mice, and the specific steps are as follows: the mice used in this animal experiment are 6-week-old db / db mice (BKS-Lepr / em2Cd479 / Gpt). The mice are raised in a SPF environment at 25℃, 12h light / 12h dark, and adapted for one week. Then, the db / db mice are used as the model mice for subsequent T2DM research to carry out related animal experiments. All the mice are randomly divided into groups, of which 10 db / db mice are divided into two groups, 10 wild type mice are used as the model control group, 8 mice are used as the positive control drug group, 6 mice are used as the low concentration group of the compound, 7 mice are used as the medium concentration group of the compound, and 7 mice are used as the high concentration group of the compound. The administration interval is one day, the test compound is injected intraperitoneally, the low concentration group of the compound refers to 5mg / kg / day, the medium concentration group of the compound refers to 10mg / kg / day, the high concentration group of the compound refers to 20mg / kg / day, and the positive control drug group (10mg / kg / day) is administered orally. After administration, the random blood glucose and fasting blood glucose (overnight fasting for 12h) are detected once a week, the IPGTT is detected once every two weeks, the fasting blood glucose is recorded before intraperitoneal injection of glucose, and then the blood glucose is monitored and recorded at 30, 60, 90 and 120 minutes after injection of glucose, respectively. The ability of the mice to decompose and metabolize glucose after four weeks of administration is detected, and the hypoglycemic effect is comprehensively evaluated. Finally, the related blood, liver, spleen and other tissues are collected for subsequent related research. The results are shown in Figure 8 、 Figure 9

[0138] The experimental results show that:

[0139] (1) The compound of the present application has weak activation ability on PPARγ. Taking rosiglitazone, a diabetes treatment drug, as the positive control, the compound of the present application only weakly activates PPARγ at a concentration of 10μM, which indicates that the compound of the present application only produces weak side effects.

[0140] (2) The compound synthesized in the present application has strong binding force with PPARγ. Taking rosiglitazone, a diabetes treatment drug, as the positive control, it is shown that the compound synthesized in the present application binds to PPARγ well at a concentration of 10μM.

[0141] (3) The compound synthesized in the present application has the ability to inhibit adipocyte differentiation. Taking rosiglitazone, a diabetes treatment drug, as the positive control (the test value is 41.86%), it is shown that the adipocyte differentiation rate of the compound synthesized in the present application is only between 6.68-15.75% at a concentration of 10μM, which is much lower than that of rosiglitazone.

[0142] ​(4) The synthesized compound of the present application only weakly activates the expression of PPARγ downstream adipogenic genes, can selectively increase the expression of insulin-sensitive genes such as Adiponectin, and inhibit CDK5-mediated phosphorylation of PPARγ-Ser 273, thereby exerting good hypoglycemic effect.

[0143] (5) YGL-12 weakly induces the differentiation ability of 3T3-L1 preadipocytes and the expression of related adipogenic genes such as PPARγ, aP2, CD36, C / EBPα and Fasn, and does not cause body weight and major tissue weight increase.

[0144] The above description of the embodiments is only used to help understand the technical solutions of the present application and its core idea. It should be pointed out that for those skilled in the art, without departing from the principles of the present application, the present application can be improved and modified in several ways, and these improvements and modifications also fall within the protection scope of the claims of the present application.

Claims

1. The use of a benzothiophene ketone derivative in the preparation of an antidiabetic drug, characterized in that: The chemical structure of the benzothiophene ketone derivative is shown in formula (I): (I) Wherein, R is any one of the following groups: C3-C 10 Alkanes and N-benzyl-2-chloroformamide.

2. The use according to claim 1, characterized in that The benzothiophene ketone derivative is one of the following compounds: (4-(Hexyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(octyloxy)phenyl)methanone; N-Benzyl-2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carboxyl)phenoxy)acetamide; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-propoxyphenyl)methanone; (4-Butoxyphenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyloxy)phenyl)methanone; (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(nonyloxy)phenyl)methanone; (4-(Decyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (4-(heptane-3-oxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(3-phenylpropyloxy)phenyl)methanone; (4-(Cyclohexylmethoxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone.

3. The use according to claim 1 or 2, characterized in that The preparation method of the benzothiophene ketone derivative comprises the following steps: compound 1 and compound 2 are subjected to a Friedel-Crafts acylation reaction to obtain intermediate A, intermediate A is subjected to a demethylation reaction to obtain intermediate B, intermediate B is subjected to an alkylation reaction with a halogenated alkane or N-benzyl-2-chloroformamide to obtain intermediate C, and the intermediate is subjected to a demethylation reaction to finally obtain product D, i.e., a benzothiophene ketone derivative, wherein R is any one of the following groups: C3-C 10 Alkanes and N-benzyl-2-chloroformamide; the reaction formula is shown in formula (II): Formula (II).

4. The use according to claim 3, characterized in that The molar ratio of compound 1 to compound 2 is 1:5, and the Friedel-Crafts acylation reaction conditions are 1-2 h at room temperature; the molar ratio of intermediate B to halogenated alkane or N-benzyl-2-chloroformamide is 1:2, and the reaction is carried out at room temperature for 12 h.

5. The use according to claim 1, characterized in that The anti-diabetic drug is an anti-type II diabetes drug targeting PPARγ.

6. An antidiabetic drug, characterized in that: Contains an effective amount of a benzothiophene ketone derivative or 2-(4-hydroxyphenyl)-3-{4-[2-(1-piperidinyl)ethoxy]benzoyl}-6-hydroxybenzo[b]thiophene hydrochloride as an active ingredient and a pharmaceutically acceptable carrier; the benzothiophene ketone derivative is one of the following compounds: (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(octyloxy)phenyl)methanone; N-Benzyl-2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carboxyl)phenoxy)acetamide; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(pentyloxy)phenyl)methanone; (4-(heptyloxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(nonyloxy)phenyl)methanone; (4-(heptane-3-oxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone; (6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)(4-(3-phenylpropyloxy)phenyl)methanone; (4-(Cyclohexylmethoxy)phenyl)(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophen-3-yl)methanone.

7. The antidiabetic drug according to claim 6, characterized in that The anti-diabetic drug is an anti-type II diabetes drug targeting PPARγ.

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