CN107235842B

CN107235842B - Phenylpropanoate derivative and preparation method and application thereof

Info

Publication number: CN107235842B
Application number: CN201710561999.0A
Authority: CN
Inventors: 陈福欣; 侯彬彬; 龚频; 文和; 郑超; 李刚; 周安宁
Original assignee: Xian University of Science and Technology
Current assignee: Xian University of Science and Technology
Priority date: 2017-07-11
Filing date: 2017-07-11
Publication date: 2021-02-26
Anticipated expiration: 2037-07-11
Also published as: CN107235842A

Abstract

The invention discloses a phenylpropionate derivative, a preparation method and an application thereof, wherein the structure of the phenylpropionate derivative is shown as a formula I:

wherein R is¹、R²Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl, alkyl; r³Is H, CH₃、i‑Pr、t‑Bu、CF₃Or OCH₃. The rat diabetes model induced by the OGTT and STZ of a normal mouse shows that the derivative has obvious hypoglycemic activity, can reduce the blood sugar of a rat of the diabetes model in a dose-dependent manner, has a certain protection effect on a mouse with high-fat diabetes, proves the hypoglycemic effect and the lipid-lowering effect of the derivative, and can be used for preparing related medicaments for the adjuvant therapy of diabetes, particularly obesity-type diabetes and diabetic complications.

Description

Phenylpropanoate derivative and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmacy, and relates to a phenylpropionate derivative and application thereof in sugar reduction and kidney protection, in particular to application of the derivative in preparation of relevant medicines for resisting diabetes and complications thereof and the like.

Background

Diabetes mellitus is a chronic disease caused by disorders of the endocrine system. It is expected that the population suffering from diabetes worldwide will increase from 2.85 to 5.66 billion in 2013 by 2030. The diabetes mellitus period has long action time, is characterized by hyperglycemia, and is easy to cause damage to other organs such as eyes, kidneys, blood vessels, heart and the like. It has the phenomena of high morbidity, high mortality, high disability rate, low awareness rate, low treatment rate and low control rate. Diabetes is classified into type 1 diabetes and type 2 diabetes. Type 1 clinically manifests as polydipsia, diuresis, polyphagia and emaciation, type 2 fatigue and weakness. Type 1 diabetes is most common in teenagers, generally less than 30 years old, and type 2 diabetes is common in middle-aged and elderly people, and accounts for over 70% of patients with diabetes.

The research on the existing sugar-reducing medicines such as sulfonylureas, thiazolidinediones, alpha-glucosidase inhibitors, DPP-IV inhibitors, SGLT2 inhibitors and the like mainly focuses on the sugar-reducing effect, has little focus on the fat-reducing effect, and needs to develop new diabetes mellitus treatment medicines with treatment effects on diabetic complications, obese diabetes mellitus and the like.

Disclosure of Invention

The invention aims to provide a phenylpropionate derivative and a preparation method and application thereof.

In order to achieve the purpose, the invention adopts the following technical scheme:

a phenylpropionate derivative, the structure of which is shown in formula 1:

wherein R is¹Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl or alkyl (alkyl preferably having 5 or less carbon atoms); r²Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl or alkyl (alkyl preferably having 5 or less carbon atoms); r is selected from

R³Is H, CH₃、i-Pr、t-Bu、CF₃Or OCH₃。

R is selected from

The structure of the phenylpropionic acid ester derivative is shown as a formula IV, a formula V, a formula VI or a formula VII:

the preparation method of the phenylpropionate derivative comprises the following steps:

the phenylpropionic acid or the phenylpropionic acid derivative and the 1- (halogenated methyl) naphthalene derivative or the 2- (halogenated methyl) naphthalene derivative are subjected to condensation reaction, and then the phenylpropionic acid derivative is obtained through extraction, washing, drying and column chromatography purification in sequence.

The preparation method of the phenylpropionate derivative specifically comprises the following steps:

1) dissolving a raw material A in an organic solvent to obtain a solution I, wherein the raw material A is phenylpropionic acid or a phenylpropionic acid derivative;

2) adding alkali into the solution I, and stirring for 0.5-1 h at 1-30 ℃ (generally at room temperature) to obtain a solution II;

3) sequentially adding iodized salt and the raw material B into the solution II to obtain reaction liquid; adding iodized salt and adding a raw material B at an interval of 10-20 min, wherein the raw material B is a 1- (halogenated methyl) naphthalene derivative or a 2- (halogenated methyl) naphthalene derivative, heating the reaction solution to 40-80 ℃, and reacting for 3-12 hours;

4) after the reaction is finished (the extraction can be carried out after cooling, or the extraction can be carried out directly), firstly, extracting by using a mixed solution of saturated sodium chloride aqueous solution and ethyl acetate, wherein the volume ratio of the saturated sodium chloride aqueous solution to the ethyl acetate in the mixed solution is 1: 1-2: 1 (the use amount of the mixed solution is calculated by 1mmol of the raw material A and is 9-15 mL), continuously extracting the water phase obtained by extraction by using ethyl acetate (calculated by 1mmol of the raw material A and 2-5 mL each time and 2-3 times), then washing the organic phase obtained by extraction, drying anhydrous sodium sulfate after washing, and then carrying out rotary evaporation to remove the solvent to obtain a solid crude product, wherein the crude product is purified by column chromatography to obtain the phenylpropionate derivative.

The organic solvent is selected from aprotic organic solvents (the dosage is 1-2 mL calculated by 1mmol of the raw material A), preferably organic solvents containing N with a boiling point higher than that of water, such as DMF (dimethylformamide), DMSO (dimethyl sulfoxide), N-methylpyrrolidone (for the preferred reason, 1) and no active H; 2) the solvent is relatively polar); the base is selected from carbonate or bicarbonate; the iodine salt is selected from cesium iodide, potassium iodide, sodium iodide or lithium iodide.

The structure of the phenylpropionic acid derivative is shown as a formula 2:

wherein R is¹Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl or alkyl (alkyl preferably having 5 or less carbon atoms), R²Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl or alkyl (alkyl preferably having 5 or less carbon atoms);

the structure of the 1- (halogenated methyl) naphthalene derivative or the 2- (halogenated methyl) naphthalene derivative is shown as a formula 3:

wherein R is³Is H, CH₃、i-Pr、t-Bu、CF₃Or OCH₃X is Cl, Br or I, and CH₂X-has two different substituent positions of alpha and beta (namely, 1 position and 2 position), namely, the 1- (halomethyl) naphthalene derivative has the structure

Or the 2- (halomethyl) naphthalene derivative has the structure

According to the material quantity ratio, the dosage ratio of the raw material A to the alkali is 1: 1-1: 3, the dosage ratio of the raw material A to the iodized salt is 1: 1-1: 4, and the dosage ratio of the raw material A to the raw material B is 1: 1-1: 3.

The specific steps of the washing are as follows: and (3) sequentially washing the organic phase obtained by extraction with a saturated copper sulfate aqueous solution (calculated by 1mmol of the raw material A, DMF, DMSO, N-methylpyrrolidone and the like are removed for 3-6 times at a time) and a saturated sodium chloride aqueous solution (calculated by 1mmol of the raw material A, DMF, DMSO, N-methylpyrrolidone and the like are removed for 1.5-6 mL, 1-2 times at a time), or washing the organic phase obtained by extraction with only a saturated sodium chloride aqueous solution (calculated by 1mmol of the raw material A, 2-6 mL, 1-2 times at a time) when an organic solvent with a boiling point higher than that of water containing N is not used, so as to remove residual water-soluble impurities.

In the column chromatography, gradient elution is carried out according to the volume ratio of petroleum ether to ethyl acetate of 10: 1-2: 1, and the specific conditions of the column chromatography are as follows:

the type of the column chromatography silica gel is 100-300 meshes; the dosage is 10-100 times of the mass of the solid crude product; adding column chromatography silica gel twice (by mass ratio) to the crude product, uniformly stirring, and then carrying out column chromatography;

the gradient elution was:

gradient I, wherein petroleum ether and ethyl acetate are 10:1 (volume ratio), and the total volume (unit: ml) is 10-20 times of the mass (unit: g) of the solid crude product;

gradient II, petroleum ether and ethyl acetate are 5:1 (volume ratio), and the total volume (unit: ml) is 4-15 times of the mass (unit: g) of the solid crude product;

gradient III, wherein the petroleum ether and the ethyl acetate are 2:1 (volume ratio), and the total volume (unit: ml) is 5-10 times of the mass (unit: g) of the solid crude product;

collecting 4-5 mL of each test tube, identifying the same chromatographic peak by TLC (GF254), collecting, combining, and removing the solvent by rotary evaporation at-0.08 MPa and 40 ℃.

The multiple of the total volume to the mass of the solid crude product varies depending on the specific structure of the phenylpropionate derivative to be prepared, thereby forming the above multiple range.

The phenylpropionic acid ester derivative can be applied to the preparation of medicines for preventing and treating diabetes, medicines for assisting in treating diabetic complications and medicines for reducing blood sugar.

The application of the phenylpropionate derivative in preparing lipid-lowering medicines is provided.

The application of the phenylpropionate derivative in preparing medicines for preventing and treating obesity-type diabetes and hyperlipidemia-type diabetes is provided.

The invention has the beneficial effects that:

the synthetic method of the phenylpropionic acid ester derivative is simple, synthetic raw materials are easy to obtain, the remarkable hypoglycemic activity of the phenylpropionic acid ester derivative is verified through in-vivo experiments of diabetes model animals, and the lipid-lowering effect of the phenylpropionic acid ester derivative is preliminarily proved through detection of biochemical indexes such as total cholesterol, triglyceride and the like. The pharmacokinetic experiment preliminarily proves the good pharmacokinetic property of the compound. The phenylpropionate derivative can be used as a medicinal active ingredient to prepare medicaments for resisting diabetes, improving lipid metabolism and assisting in treating diabetic complications.

On the basis of the existing synthesis process, the invention optimizes the synthesis route and process parameter conditions, improves the yield, is easy to separate and purify the product, simultaneously expands the application range of the substrate, and has better applicability to substrates substituted by various functional groups.

Detailed Description

The present invention will be described in detail with reference to examples.

A series of phenylpropionic acid ester derivatives are synthesized and screened, and biological activity data show that the compounds have very good in-vivo hypoglycemic effect and ideal pharmacokinetic property, so that new application is provided for the compounds, and the compounds have very important significance particularly in the aspect of medicine and pharmacology.

The series of phenylpropionate derivatives are obtained by condensation reaction of phenylpropionic acid (or phenylpropionic acid derivatives) and 1 or 2- (halogenated methyl) naphthalene derivatives.

The structure of the phenylpropionate derivative is shown as a formula I:

wherein R is¹、R²Is H, OH, OCH₃Isopentenyl, 3-methylbut-2-enyl or alkyl; r³Is H, CH₃、i-Pr、t-Bu、CF₃Or OCH₃And there are two different substituent positions alpha and beta (i.e., positions 1 and 2).

The invention adopts the OGTT experiment of a normal mouse and the STZ induced II-type rat diabetes model to evaluate the influence of the phenylpropionate derivative on the glucose metabolism in the diabetes model, and compared with other evaluation models, the invention can more truly reflect the glucose metabolism environment of a diabetic patient; meanwhile, pharmacokinetic experimental data show that part of compounds have very good drug properties. Biochemical test data show that part of compounds also have obvious lipid-lowering activity, so the phenylpropionate derivatives are particularly suitable for further research on obese diabetes.

Synthesis examples

EXAMPLE 1 Synthesis of Dihydrocaffeic acid-2-naphthylmethyl ester (abbreviated as PN1, shown in formula II)

1) 1.10mmol of dihydrocaffeic acid was weighed out, dissolved in 1.5mL of DMSO and 1.32mmol of Na was added₂CO₃After stirring at room temperature for 0.5h, 1.10mmol of potassium iodide and 1.20mmol of 2- (chloromethyl) naphthalene were added in succession. The reaction mixture was stirred at 60 ℃ for 6h (TLC check reaction complete).

2) After the reaction is finished, cooling to room temperature, adding 5mL of saturated sodium chloride solution and 5mL of ethyl acetate, uniformly mixing, and standing for layering; the aqueous phase was extracted twice with ethyl acetate (3 mL. times.2), the organic phases were combined and washed with saturated copper sulfate solution (3 mL. times.4) and saturated sodium chloride solution (2 mL. times.2), dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation (-0.08MPa, 45 ℃ C.) to give the crude solid product.

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I is 20 times, gradient II is 5 times, and gradient III is 5 times), and then dihydrocaffeic acid-2-naphthyl methyl ester (PN1220.4mg, 65% yield) is obtained. Number of structural identificationsThe following is provided: LC-MS:323.1(M + 1)⁺)。¹HNMR(400MHz,DMSO-d6)δ:8.85(s,2H),8.01-7.90(m,3H),7.61-7.51(m,3H),7.48(t,J＝7.6Hz,1H),6.73-6.64(m,2H),6.52(dd,J＝8.2,2.0Hz,1H),5.56(s,2H),2.71(d,J＝7.4Hz,2H),2.61(d,J＝7.4Hz,2H)。

Example 2 Synthesis of 1-naphthylmethyl p-hydroxyphenylpropionate (abbreviated as PN2, shown in formula III)

1) 1.10mmol of p-hydroxyphenylpropionic acid are weighed out, dissolved in 1.5mL of acetone and 2.2mmol of NaHCO are added₃After stirring at room temperature for 1h, 2.2mmol of lithium iodide and 2.2mmol of 1- (bromomethyl) naphthalene were added in succession. The reaction mixture was stirred at 40 ℃ for 12h (TLC check reaction complete).

2) After the reaction is finished, cooling to room temperature, adding 10mL of saturated sodium chloride solution and 5mL of ethyl acetate, uniformly mixing, and standing for layering; the aqueous phase was extracted twice with ethyl acetate (3 mL. times.2), the organic phases were combined, washed with saturated sodium chloride solution (2 mL. times.2), dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation (-0.08MPa, 45 ℃ C.) to give the crude solid product.

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I is 20 times, gradient II is 5 times, and gradient III is 5 times), to obtain p-hydroxyphenylpropionic acid-1-naphthyl methyl ester (PN2229.6 mg, 68% yield). The structure identification data is as follows: LC-MS:307.1(M + 1)⁺)。¹HNMR(400MHz,DMSO-d6)δ:9.22(s,1H),7.91(dd,J＝8.9,4.0Hz,3H),7.83(s,1H),7.53(p,J＝4.9Hz,2H),7.43(d,J＝8.4Hz,1H),7.01(d,J＝8.1Hz,2H),6.65(d,J＝8.0Hz,2H),5.24(s,2H),2.78(t,J＝7.4Hz,2H),2.66(t,J＝7.4Hz,2H)。

EXAMPLE 3 Synthesis of 1-naphthalene- (5-isopropyl) -methyl dihydrocaffeate (abbreviated as PN3, shown in formula IV)

1) 1.10mmol of dihydrocaffeic acid was weighed out, dissolved in 1.5mL of DMF and 3.3mmol of KHCO were added₃After stirring at room temperature for 1h, 3.3mmol of sodium iodide and 1.20mmol of 1- (chloromethyl) - (5-isopropyl) -naphthalene were added in succession. The reaction mixture was stirred at 80 ℃ for 3h (TLC check for reaction completion).

2) After the reaction is finished, cooling to room temperature, adding 5mL of saturated sodium chloride solution and 5mL of ethyl acetate, uniformly mixing, and standing for layering; the aqueous phase was extracted twice with ethyl acetate (3 mL. times.2), the organic phases were combined and washed with saturated copper sulfate solution (4 mL. times.4) and saturated sodium chloride solution (2 mL. times.2), dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation (-0.08MPa, 45 ℃ C.) to give the crude solid product.

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I of 20 times, gradient II of 10 times and gradient III of 5 times) to obtain dihydrocaffeic acid-1-naphthalene- (5-isopropyl) -methyl ester (PN3180.6mg, 45% yield). The structure identification data is as follows: LC-MS:365.2(M + 1)⁺)。¹H NMR(400MHz,DMSO-d6)δ8.94(s,1H),8.88(s,1H),7.92(m,3H),7.80(s,1H),7.54(m,1H),7.45(d,J＝8.3Hz,1H),6.66(m,2H),6.53(d,J＝8.3,1H),5.53(s,2H),3.19(m,1H),2.72(d,J＝7.4Hz,2H),2.60(d,J＝7.4Hz,2H),1.36(d,J＝8.0Hz,6H)。

Example 43 Synthesis of 2-naphthalen- (5-methyl) -methyl (3-methoxy-4-hydroxyphenyl) -propionate (abbreviation PN4, formula V)

1) 1.10mmol of 3- (3-methoxy-4-hydroxyphenyl) -propionic acid are weighed out, dissolved in 2mL of DMSO and 1.1mmol of Li are added₂CO₃After stirring at room temperature for 1h, 4.4mmol of cesium iodide and 2.2mmol of 2- (chloromethyl) - (5-methyl) -naphthalene were added in succession. The reaction mixture was stirred at 60 ℃ for 10h (TLC check reaction complete).

2) After the reaction is finished, cooling to room temperature, adding 5mL of saturated sodium chloride solution and 8mL of ethyl acetate, uniformly mixing, and standing for layering; the aqueous phase was extracted twice with ethyl acetate (3 mL. times.2), the organic phases were combined and washed with saturated copper sulfate solution (3 mL. times.3) and saturated sodium chloride solution (2 mL. times.2), dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation (-0.08MPa, 45 ℃ C.) to give the crude solid product.

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I15-fold, gradient II 10-fold, gradient III 10-fold) gave 3- (3-methoxy-4-hydroxyphenyl) -propionic acid-2-naphthalen- (5-methyl) -methyl ester (PN 4201 mg, 52% yield). The structure identification data is as follows: LC-MS:351.2(M + 1)⁺)。¹HNMR(400MHz,CDCl3)δ:8.10(s,1H),8.03-7.93(m,3H),7.64-7.53(m,2H),7.46(t,J＝7.6Hz,1H),6.84(d,J＝2Hz,1H),6.65(m,2H),5.25(s,2H),3.83(s,3H),2.90(d,J＝7.4Hz,2H),2.69(d,J＝7.4Hz,2H),2.53(s,3H)。

Example Synthesis of 53- (4-hydroxy-3, 5-diisopentenylphenyl) -propionic acid-1-naphthalene- (5-trifluoromethyl) -methyl ester (abbreviated as PN5, formula VI)

1) 1.10mmol of 3- (4-hydroxy-3, 5-diisopentenylphenyl) -propionic acid are weighed out, dissolved in 2mL of N-methylpyrrolidone and 1.8mmol of Cs are added₂CO₃After stirring at room temperature for 1h, 1.20mmol of cesium iodide and 3.20mmol of 1- (chloromethyl) - (5-trifluoromethyl) -naphthalene were added in succession. The reaction mixture was stirred at 60 ℃ for 8h (TLC check reaction complete).

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I is 20 times, gradient II is 10 times, ladderDegree III 5 fold) to give 3- (4-hydroxy-3, 5-diisopentenylphenyl) -propionic acid-1-naphthalen- (5-trifluoromethyl) -methyl ester (PN 5400 mg, 71% yield). The structure identification data is as follows: LC-MS:511.2(M + 1)⁺)。¹HNMR(400MHz,DMSO-d6)δ:8.06(s,1H),8.03-7.98(m,2H),7.69-7.49(m,3H),7.43(t,J＝7.6Hz,1H),6.83-6.73(m,2H),5.55(s,2H),5.29-5.34(m,2H),3.21(d,J＝7.2Hz,4H),2.92(d,J＝7.4Hz,2H),2.71(d,J＝7.4Hz,2H),1.79(s,6H),1.77(s,6H)。

Example Synthesis of 63- (4-hydroxy-3-isopentenylphenyl) propanoic acid-1-naphthalen- (5-methoxy) -methyl ester (abbreviated as PN6, shown in formula VII)

1) 1.10mmol of 3- (4-hydroxy-3-isopentenylphenyl) propionic acid was weighed out, dissolved in 1.5mL of DMSO, and 1.32mmol of KHCO was added₃After stirring at room temperature for 1h, 1.20mmol of cesium iodide and 2.60mmol of 1- (chloromethyl) - (5-methoxy) -naphthalene were added in succession. The reaction mixture was stirred at 50 ℃ for 5h (TLC check reaction complete).

2) After the reaction is finished, cooling to room temperature, adding 5mL of saturated sodium chloride solution and 10mL of ethyl acetate, uniformly mixing, and standing for layering; the aqueous phase was extracted twice with ethyl acetate (3 mL. times.2), the organic phases were combined and washed with saturated copper sulfate solution (3 mL. times.4) and saturated sodium chloride solution (2 mL. times.2), dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation (-0.08MPa, 45 ℃ C.) to give the crude solid product.

3) The crude product was purified by column chromatography, column chromatography (the total volume times the mass of the solid crude product: gradient I15 times, gradient II 15 times, gradient III 5 times) to give 1-naphthalen- (5-methoxy) -methyl 3- (4-hydroxy-3-isopentenylphenyl) propanoate (pn6187.1mg, 42% yield). The structure identification data is as follows: LC-MS 405.2(M + 1)⁺)。¹HNMR(400MHz,DMSO-d6)δ:8.19(s,1H),7.75-7.54(m,2H),7.20-7.09(m,2H),6.92-6.82(m,2H),6.83-6.73(m,2H),6.64(s,1H),5.51(s,2H),5.32(m,1H),3.80(s,3H),3.21(d,J＝7.2Hz,2H),2.91(d,J＝7.4Hz,2H),2.70(d,J＝7.4Hz,2H),1.79(s,3H),1.77(s,3H)。

(II) oral glucose tolerance test in Normal mice

Assay methods reference standard methods reported, briefly described as: before the experiment, the C57BL/6 mice in each half of the males and females are fasted for 12h, and are subjected to intragastric administration at the glucose dose of 2g/kg (concentration of 0.4g/mL) body weight, and 30min before the intragastric administration (intragastric administration). Blood Glucose (BG) was measured with a glucometer at 0, 0.5, 1, 1.5, 2h after gavage with glucose, respectively. Each group of experimental animals contained 7 animals, 56 animals in total.

Mode of administration

(1) Blank group: physiological saline (20mL/kg) in equal volume with the other groups;

(2) PN group (group administered with phenylpropionate derivative, same molarity of PN1-PN6, dissolution with physiological saline, small amount of DMSO as cosolvent): the medicine is administrated by gastric lavage according to 20 mu mol/kg;

(3) positive group: dissolving metformin in ultrapure water to a final concentration of 10g/L, and performing intragastric administration according to 20 mu mol/kg;

tail vein blood sampling, and determination by an ease type Roche glucometer; the experimental data of each group were averaged.

The test results and data analysis (see table 1 for results) show that: (1) compared with the blank group, the PN group can reduce the blood sugar to different degrees; (2) the phenylpropionate derivatives prepared in the above examples had slightly lower or close hypoglycemic ability to metformin compared to the positive group, wherein the hypoglycemic ability of PN5 and PN6 was very close to that of metformin.

Therefore, for a normal mouse model, the prepared phenylpropionate derivative has obvious hypoglycemic capability.

TABLE 1 results of oral glucose tolerance test in Normal mice

(III) diabetes model of STZ-induced rat and hypoglycemic effect of drug

The model construction and drug screening method reference literature report standard method, briefly described as: SD mice are taken and divided into a model group and a normal control group by a drawing method. After the model group is fasted for 12 hours without water prohibition, the left lower abdominal cavity is injected with STZ 40mg/kg (0.01mol/L, pH 4.2 citric acid buffer solution is used as a solvent) for 1 time/d and continuously for 5 days. The normal control group is fasted and is not forbidden to inject citric acid buffer solution with equal volume for 12 hours, 1 time/d and 5 days continuously. Within 1 week after the last injection, blood glucose was measured by cutting off the tail and taking blood every day. Diabetic rats were treated with continuous 3 days of blood glucose >16.7mmol/L and drug screening experiments were performed on day 4.

Drug screening experiments: rats successfully modeled were fasted for 12h and gavaged with a 2g/kg (0.4 g/mL) body weight glucose dose with concurrent gavage. Measuring Blood Glucose (BG) with a glucometer at 0, 0.5, 1, 1.5, 2h after administration; the area under the curve (AUC) at time of administration reflects glucose tolerance.

Mode of administration

(1) Blank control group (blank group for short): physiological saline (20mL/kg) in equal volume to the other groups; this group was different from the normal control group, and the blank control group was also a model animal, but was not administered, and was given physiological saline only.

(2) Phenylpropionate derivatives experimental group (PN group for short, concentration of PN2-PN5 is the same): the derivative is administrated by intragastric administration according to 20 mu mol/kg;

(3) positive control drug group (positive group for short): dissolving metformin in ultrapure water to a final concentration of 10g/L, and performing intragastric administration according to 20 mu mol/kg;

5 experimental animals in each group, and the blood sugar determination and data processing method are the same as the second method.

The test results and data analysis (see table 2) show that: the sugar-reducing ability of PN5 and PN6 in the STZ-induced SD rat diabetes model is slightly higher than that of metformin, the regulation of blood sugar in vivo is more gradual, and the method is favorable for further preclinical research of antidiabetic drugs.

TABLE 2 STZ-induced results of the SD rat diabetic model

(IV) Biochemical detection of lipid lowering

The model construction and drug screening method reference literature report standard method, briefly described as: 42 Kunming mice were selected and classified into model group (n ═ 28) and blank control group (n ═ 7) by the extraction method. The model groups are all fed with high-fat high-sugar feed, and the feeding amount is as follows: the mice had free food, about 6g a day, without water. The high-fat high-sugar feed comprises the following components in parts by mass: 18% of lard, 20% of cane sugar, 3% of egg yolk and 59% of daily maintenance type feed for experimental mice. After 8 weeks of feeding, each group of mice was fasted for 12 hours, and Triglyceride (TG), Total Cholesterol (TC), high density lipoprotein (HDL-C), and low density lipoprotein (LDL-C) were measured by a full-automatic biochemical analyzer (HITACHI 7600).

The administration mode is as follows:

(1) blank control group: feeding normal feed, and simultaneously performing intragastric administration and equal volume of normal saline (20mL/kg) of other groups;

(2) model control group: feeding high-fat high-sugar feed, and simultaneously performing intragastric administration and equal volume of physiological saline (20mL/kg) of other groups;

(3) protection group (PN5 and PN6 group): feeding high-fat high-sugar feed, and simultaneously performing intragastric administration (normal saline is dissolved, and 5% Tween 80 is used for helping dissolution), wherein the dosage is 20 mu mol/kg/day, and the total period is 8 weeks;

(4) positive group: feeding high-fat high-sugar feed, dissolving atorvastatin (Lipitor) in ultrapure water, assisting in dissolving with DMSO to obtain a final concentration of 10mg/mL, and performing intragastric administration at a dose of 20 μmol/kg/day;

TABLE 3 Biochemical test results for lipid lowering model mice

Test results and data analysis (see Table 3 for results)

(1) Compared with a model group, PN5 and PN6 have a protective effect on various biochemical indexes of blood fat, and can effectively reduce blood fat; wherein, the content of TG can be reduced to normal level; it has a better normalising effect on TC relative to atorvastatin;

(2) compared with a model group, PN5 and PN6 groups have very good protection effect on LDL-C, and the content of LDL-C can be greatly reduced; meanwhile, the protective effect on HDL-C is basically the same.

The lipid-lowering effect of PN1-PN 4 is inferior to that of PN5 and PN 6.

The phenylpropionic acid ester derivative disclosed by the invention is shown to have obvious hypoglycemic activity through a rat diabetes model induced by OGTT and STZ of a normal mouse, can reduce the blood sugar of the rat of the diabetes model in a dose-dependent manner, has a certain protection effect on a mouse with high-fat diabetes mellitus, proves the hypoglycemic effect and the lipid-lowering effect of the derivative, and can be used for preparing related medicines for diabetes mellitus, particularly obesity-type diabetes mellitus.

(V) pharmacokinetic experiments

The pharmacokinetic experiment adopts a method reported in the literature, and is briefly described as follows: taking a diabetic rat successfully modeled by the STZ, and administrating PN6(5mg/kg) through intragastric administration; blood is taken at 0min, 15min, 30min, 45min, 60min, 90min, 120min and 150min, 600 muL of methanol is added into 150 muL of plasma for protein removal, after SPE purification, UPLC-MS/MS detects the plasma concentration of PN6 in a sample (MRM scanning mode, ion pair is 405.2-188.1m/z), and pharmacokinetic properties are predicted in a two-chamber mode (Table 4).

TABLE 4 pharmacokinetic Properties of PN6(5mg/kg intragastric administration)

Preliminary pharmacokinetic experiments show that the in vivo half-life period of the compound PN6 is 19.3h, the plasma clearance rate is lower and is 17.2L/h/kg, the pharmacokinetic property is ideal, and the druggability is good.

Compared with the clinical hypoglycemic drugs, the phenylpropionate derivative prepared by the invention has a remarkable hypoglycemic effect, and even if the hypoglycemic capability of the phenylpropionate derivative is slightly lower than that of metformin, the phenylpropionate derivative has a certain relieving effect on the original hyperglycemic phenomenon. The phenylpropionic acid ester derivative prepared by the invention can be used as a lead compound of a medicament for preventing and treating diabetes.

Through a blood sugar reducing model and a blood fat reducing experiment, a compound PN6 is preliminarily screened, has a certain blood fat reducing effect while reducing blood sugar under the experimental condition, has better medicament performance, and can be used as a blood sugar reducing medicament, in particular to a lead compound for hyperlipemia and diabetes mellitus.

Claims

1. A phenylpropionate derivative characterized in that: the structure of the phenylpropionate derivative is shown as a formula 1:

wherein R is₁Is isopentenyl; r₂Is H or isopentenyl; r is selected from

R₃Is CF₃Or OCH₃。

2. The phenylpropionate derivative according to claim 1, wherein: r is selected from

3. A phenylpropionate derivative according to claim 1 or 2, wherein: in the formula 1, the structure of the phenylpropionic acid ester derivative is shown as a formula VI or a formula VII:

4. the process for the preparation of a phenylpropionate derivative according to claim 1, wherein: the method comprises the following steps:

the phenylpropionic acid derivative and 1- (halogenated methyl) naphthalene derivative or 2- (halogenated methyl) naphthalene derivative are subjected to condensation reaction, and then are subjected to extraction, washing, drying and column chromatography purification in sequence to obtain the phenylpropionic acid ester derivative.

5. The method of claim 4, wherein: the preparation method of the phenylpropionate derivative specifically comprises the following steps:

1) dissolving a raw material A in an organic solvent to obtain a solution I, wherein the raw material A is a phenylpropionic acid derivative;

2) adding alkali into the solution I, and stirring for 0.5-1 h to obtain a solution II;

4) after the reaction is finished, extracting with a mixed solution of a saturated sodium chloride aqueous solution and ethyl acetate, wherein the volume ratio of the saturated sodium chloride aqueous solution to the ethyl acetate in the mixed solution is 1: 1-2: 1, continuously extracting the water phase obtained by extraction with ethyl acetate, washing the organic phase obtained by extraction, drying with anhydrous sodium sulfate after washing, and then performing rotary evaporation to remove the solvent to obtain a solid crude product, wherein the crude product is purified by column chromatography to obtain the phenylpropionate derivative.

6. The method of claim 5, wherein: the organic solvent is selected from aprotic organic solvents; the base is selected from carbonate or bicarbonate; the iodine salt is selected from cesium iodide, potassium iodide, sodium iodide or lithium iodide.

7. The method of claim 5, wherein: according to the material quantity ratio, the dosage ratio of the raw material A to the alkali is 1: 1-1: 3, the dosage ratio of the raw material A to the iodized salt is 1: 1-1: 4, and the dosage ratio of the raw material A to the raw material B is 1: 1-1: 3.

8. The method according to claim 4 or 5, characterized in that: the specific steps of the washing are as follows: washing the organic phase obtained by extraction with a saturated copper sulfate aqueous solution and a saturated sodium chloride aqueous solution in sequence, or washing the organic phase obtained by extraction with a saturated sodium chloride aqueous solution only; in the column chromatography, gradient elution is carried out according to the volume ratio of petroleum ether to ethyl acetate of 10: 1-2: 1.

9. Use of the phenylpropionate derivative according to claim 1 for the preparation of a lipid lowering drug.