A phenylpiperazine-based β-carboline derivative, and its preparation method and application

Abstract

The present invention discloses a phenylpiperazine-based β-carboline derivative, a preparation method and an application thereof; the phenylpiperazine-based β-carboline derivative has a structural formula shown in formula (I): #imgabs0# wherein R is one or more substituents, or R does not exist; R is independently selected from halogen, C 1～6 Alkyl, C 1～6 Alkoxy, C 1～6 of halogenated alkyl, cyano, nitro, C 1～6 The present invention uses the β-carboline structure as the mother core to design and synthesize a series of phenylpiperazine-based β-carboline derivatives, which exhibit strong α-glucosidase inhibition, with an IC 50 The values ​​ranged from 4.85±0.48μM to 10.84±1.02. It can be used as an α-glucosidase inhibitor to treat and / or prevent diabetes.

Description

Technical Field

[0001] The present invention relates to the technical field of small molecule drugs, and in particular to a phenylpiperazine-based β-carboline derivative, a preparation method thereof, and an application thereof. Background Art

[0002] Diabetes has become a global problem, seriously endangering human health. Despite increased health awareness, the incidence of diabetes continues to rise. Failure to effectively and promptly control blood sugar levels can lead to a series of functional disorders and damage in the human body, seriously threatening the patient's health and safety. Type 2 diabetes is the most common, accounting for over 90% of the prevalence. Type 2 diabetes, also known as non-insulin-dependent diabetes mellitus (NIDDM), is a chronic metabolic disease caused by insulin dysfunction or insulin resistance, primarily due to the interaction of genetic and environmental factors, such as gene mutations and dietary habits.

[0003] There are many oral hypoglycemic drugs currently on the market, which act on different targets through different pathways. Sulfonylureas (glibenclamide, glimepiride and mesylate) are insulin secretagogues that can activate voltage-gated Ca 2+ Channels are used to stimulate pancreatic beta cells to release insulin. Biguanides, such as metformin, are considered the first-line treatment for diabetes. Their mechanism of action is to lower blood sugar by inhibiting hepatic gluconeogenesis and hepatobiliary sugar output. Sodium-glucose transporter inhibitors (SGLT2) are drugs, with dapagliflozin being a representative example. They can inhibit the reabsorption of glucose, thereby controlling blood sugar and reducing calories. In addition, another way to maintain blood sugar levels is to inhibit the hydrolysis of α-glucosidase, thereby lowering postprandial blood sugar.

[0004] α-glucosidase aids digestion by hydrolyzing starch and carbohydrates into glucose and monosaccharides in the small intestine. α-glucosidase inhibitors competitively and reversibly inhibit this intestinal enzyme, preventing the hydrolysis of the non-reducing ends of oligosaccharides and reducing the release of α-glucose. This hinders the digestion and absorption of carbohydrates, thereby reducing insulin demand. In addition, α-glucosidase inhibitors can lead to a long-term, sustained increase in endogenous GLP-1, thereby stimulating the secretion of insulin and reducing the secretion of glucagon, thereby achieving the effect of controlling postprandial hyperglycemia. Currently, the types of α-glucosidase inhibitors available on the market are relatively limited, and some drugs (such as acarbose) have the problem of insufficient biological activity.

[0005] Therefore, it is necessary to develop a new compound that inhibits α-glucosidase. Summary of the Invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, in a first aspect, the present invention provides a phenylpiperazine-based β-carboline derivative that can effectively inhibit α-glucosidase, thereby being used to treat and / or prevent diabetes.

[0007] The second aspect of the present invention also provides a method for preparing phenylpiperazine-based β-carboline derivatives.

[0008] The third aspect of the present invention also provides a pharmaceutical composition.

[0009] The fourth aspect of the present invention also provides an α-glucosidase inhibitor.

[0010] The fifth aspect of the present invention also provides an application.

[0011] According to the first aspect of the present invention, a phenylpiperazine-based β-carboline derivative is provided, which has the structural formula shown in formula (I):

[0012]

[0013] Wherein, R is one or more substituents, or R does not exist; R is independently selected from halogen, C 1～6 Alkyl, C 1～6 Alkoxy, C 1～6 of halogenated alkyl, cyano, nitro, C 1～6 of an alkylsulfonyl group.

[0014] The phenylpiperazine-based β-carboline derivatives provided according to the embodiments of the present invention have at least the following beneficial effects:

[0015] The present invention uses β-carboline structure as the mother core to design and synthesize a series of phenylpiperazine β-carboline derivatives, which show strong α-glucosidase inhibition. 50 The values ​​range from 4.85±0.48 μM to 10.84±1.02. The compound can be used as an α-glucosidase inhibitor for treating and / or preventing diabetes.

[0016] According to a preferred embodiment of the present invention, R is one or more substituents, or R does not exist; R is independently selected from halogen, C 1～4 Alkyl, C 1～3 Alkoxy, C 1～3 of haloalkyl, cyano, nitro, and methylsulfonyl.

[0017] According to a preferred embodiment of the present invention, R is one or more substituents, or R does not exist; R is independently selected from halogen, methyl, tert-butyl, methoxy, trifluoromethyl, cyano, nitro, and methylsulfonyl.

[0018] According to a preferred embodiment of the present invention, the phenylpiperazine β-carboline derivative is selected from one of the following structural formulas:

[0019]

[0020]

[0021]

[0022] According to a second aspect of the present invention, there is provided a method for preparing the phenylpiperazine-based β-carboline derivatives described in the first aspect of the present invention, comprising the following steps:

[0023] Compound S4, compound S5 and a base are mixed and subjected to a condensation reaction to obtain the phenylpiperazine β-carboline derivative;

[0024] The structural formulas of compound S4 and compound S5 are shown below:

[0025]

[0026] According to a preferred embodiment of the present invention, the molar ratio of compound S4 to compound S5 is 1:(1-1.5). For example, the molar ratio of compound S4 to compound S5 can be selected from 1:1, 1:1.2, 1:1.3, 1:1.4, and 1:1.5.

[0027] According to a preferred embodiment of the present invention, the base is selected from at least one of potassium hydroxide and sodium hydroxide.

[0028] According to a preferred embodiment of the present invention, the condensation reaction is carried out in the presence of a solvent, wherein the solvent is selected from at least one of ethanol and methanol.

[0029] According to a preferred embodiment of the present invention, the condensation reaction further comprises a purification step, which includes water washing and recrystallization.

[0030] According to a preferred embodiment of the present invention, the compound S4 is prepared by the following method:

[0031] Compound S3, N-phenylpiperazine, N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)uronium hexafluorophosphate (HATU), N,N-diisopropylethylamine (DIPEA) and an organic solvent are mixed and reacted to obtain a product;

[0032] The structural formula of the compound S3 is as follows:

[0033]

[0034] According to a preferred embodiment of the present invention, the organic solvent is dimethylformamide.

[0035] According to a preferred embodiment of the present invention, the molar ratio of the compound S3 to N-phenylpiperazine is 1:(1-1.2), for example, 1:1, 1:1.1, or 1:1.2.

[0036] According to a preferred embodiment of the present invention, the molar ratio of the compound S3, HATU and DIPEA is 1: (1-1.2): (2-2.2).

[0037] According to a preferred embodiment of the present invention, the reaction further comprises a post-treatment step after completion; the post-treatment comprises: adding water for quenching, and filtering the solid to obtain compound S4.

[0038] According to a preferred embodiment of the present invention, the compound S3 is prepared by the following method:

[0039] Compound S2 was used as the raw material and hydrolyzed with LiOH to obtain compound S3;

[0040] The structural formula of the compound S2 is as follows:

[0041]

[0042] According to a preferred embodiment of the present invention, the solvent for the hydrolysis reaction is a combination of tetrahydrofuran, methanol and water.

[0043] According to a preferred embodiment of the present invention, the volume ratio of tetrahydrofuran, methanol and water is (3.0-3.2): (1.0-1.2): 1. For example, it can be 3:1:1, 3.1:1.1:1, or 3.2:1.2:1.

[0044] According to a preferred embodiment of the present invention, the temperature of the hydrolysis reaction is 30-80°C.

[0045] According to a preferred embodiment of the present invention, the compound S2 is prepared by the following method:

[0046] Compound S1 is used as the starting material and undergoes a cyclization reaction with methylglyoxal in the presence of an acid to obtain compound S2;

[0047] The structural formula of the compound S1 is as follows:

[0048]

[0049] According to a preferred embodiment of the present invention, the acid is selected from at least one of concentrated sulfuric acid, hydrochloric acid, and trifluoroacetic acid.

[0050] According to a preferred embodiment of the present invention, the molar ratio of the compound S1 to methylglyoxal is 1:(1-1.2), for example, about 1:1, 1:1.1, or 1:1.2.

[0051] According to a preferred embodiment of the present invention, the post-processing steps of the cyclization reaction are as follows:

[0052] Use NaOH solution to adjust the pH to neutral, let it stand and filter to obtain the product.

[0053] According to a third aspect of the present invention, a pharmaceutical composition is provided, comprising the phenylpiperazine-based β-carboline derivatives described in the first aspect of the present invention and pharmaceutically acceptable excipients.

[0054] According to a preferred embodiment of the present invention, the dosage form of the pharmaceutical composition is selected from tablets, capsules, oral solutions or injections.

[0055] The fourth aspect of the present invention provides an α-glucosidase inhibitor comprising the phenylpiperazine-based β-carboline derivatives described in the first aspect of the present invention.

[0056] In a fifth aspect, the present invention provides a use of the phenylpiperazine-based β-carboline derivatives, the pharmaceutical composition, or the α-glucosidase inhibitor in the preparation of a drug for treating and / or preventing diabetes.

[0057] Definitions and General Terms

[0058] “C 1-6 "alkyl" means an alkyl group having a total carbon number of 1 to 6, including C 1-6 Straight chain alkyl, C 1-6 Branched alkyl and C 3-6 The cycloalkyl group may be, for example, a straight-chain alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, a branched-chain alkyl group having 3, 4, 5 or 6 carbon atoms, or a cycloalkyl group having 3, 4, 5 or 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, methylcyclopropyl, ethylcyclopropyl, cyclopentyl, methylcyclopentyl, cyclohexyl, etc.

[0059] “C 1-6 "alkoxy" means an alkoxy group having a total carbon number of 1 to 6, including C 1-6 Straight chain alkoxy, C 3-6 Branched alkoxy and C 2-6The cycloalkoxy group can be, for example, a straight-chain alkoxy group having 1, 2, 3, 4, 5 or 6 carbon atoms, a branched-chain alkoxy group having 3, 4, 5 or 6 carbon atoms, or a cycloalkoxy group having 2, 3, 4, 5 or 6 carbon atoms, for example, a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group, etc.

[0060] The "halogen" includes any one or two or more of fluorine, chlorine, bromine and iodine.

[0061] “C 1～6 The alkyl halide and C 1-6 The definition of "alkyl" is similar, except that "C 1～6 At least one H atom in the "haloalkyl" is substituted by any halogen.

[0062] Substances that are "pharmaceutically acceptable excipients" include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances, such as phosphates; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water (including pyrogen-free water); salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silicon; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylene-polyoxypropylene-blocking polymers; lanolin; sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and Its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycol compounds such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; Ringer's solution; ethanol; phosphate buffer solution; and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate; colorants; release agents; coatings; flavorings; fragrances; preservatives and antioxidants.

[0063] Other features and advantages of the present invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0065] Figure 1 A graph showing the half-maximal inhibitory concentration of compound I-25 prepared in Example 25 of the present invention as an α-glucosidase inhibitor for α-glucosidase in vitro;

[0066] Figure 2 This is a diagram showing the enzyme kinetics of compound I-25 prepared in Example 25 of the present invention as an α-glucosidase inhibitor against α-glucosidase in vitro;

[0067] Figure 3 This is a kinetic diagram of the substrate activity of compound I-25 prepared in Example 25 of the present invention as an α-glucosidase inhibitor against α-glucosidase in vitro. DETAILED DESCRIPTION

[0068] The following are specific embodiments of the present invention, and the technical solutions of the present invention are further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0069] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0070] The reagents used in the embodiments of the present invention are as follows:

[0071] L-Tryptophan methyl ester hydrochloride (Compound S1) CAS No.: 7524-52-9, methylglyoxal CAS No.: 78-98-8, lithium hydroxide CAS No.: 1310-65-2, HATU CAS No.: 148893-10-1, DIPEA CAS No.: 7087-68-5, N-phenylpiperazine CAS No.: 92-54-6, potassium hydroxide CAS No.: 1310-58-3; all are commercially available.

[0072] Provided is a general method for preparing phenylpiperazine-based β-carboline derivatives, the reaction equation and steps are as follows:

[0073]

[0074] Synthesis of compound S2: Weigh 2 g of L-tryptophan methyl ester (compound S1; 8 mmol), add 400 mL of water, stir at room temperature, add 577 uL of methylglyoxal (9.6 mmol), and then slowly add 4 mL of concentrated sulfuric acid. React at room temperature for 24 hours and monitor with a TLC plate (PE:EA-2:1). After the reaction is completed, adjust the pH to 7 with NaOH solution, let it stand for 30 minutes, and filter out the solid to obtain the crude product S2.

[0075] Synthesis of compound S3: Weigh 500 mg of compound S2 into a 35 mL sealed tube, add 4.6 mL of a mixed solution (THF:MeOH:H0=3:1:1), add 156 mg of LiOH (2 mmol) in an ice bath, mix well, and heat to 50°C. After 2 h, monitor with a TLC plate (PE:EA=2:1). After the reaction is completed, add about 10 mL of dilute hydrochloric acid to quench the reaction, and then filter out the solid to obtain product S3.

[0076] Synthesis of compound S4: 870 mg of compound S3 (3.42 mmol) was weighed into a 100 mL two-necked flask, 1.3 g of HATU (3.42 mmol) was weighed into the flask, and 12 mL of anhydrous DMF was added. After stirring in an ice bath, 1.2 mL of DIPEA (6.84 mmol) was added. After 25 min, phenylpiperazine (3.76 mmol) was added. The atmosphere was replaced with nitrogen three times and reacted at room temperature for 24 h. The reaction was monitored by TLC plate spot plate (PE:EA=2:1). After completion of the reaction, about 10 mL of water was added to quench the reaction, and the solid was filtered to obtain product S4.

[0077] Synthesis of compounds I-1 to I-26: Weigh 120 mg of compound S4 (0.3 mmol) into a 25 mL flask, add 1 mL of ethanol and dissolve in an ice bath, then weigh 22 mg of KOH (0.39 mmol) in a small beaker, add 200 uL of water, dissolve and add dropwise to a round-bottom flask, and finally add compound S5 (0.33 mmol). After 2 hours, monitor with a TLC plate (DCM: MeOH = 20:1). After the reaction is completed, add water and filter to collect the solid, which is then recrystallized from DMF to obtain the final products I-1 to I-26.

[0078] Example 1

[0079] This example provides a phenylpiperazine β-carboline derivative I-1, whose structural formula is as follows:

[0080]

[0081] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0082] Its NMR and mass spectrometry data are as follows:

[0083] Yellow sold; yield 67%; mp298.3-299.6℃; 1H NMR (400 MHz, Chloroform-d) δ10.65(s,1H),8.78(s,1H),8.50(d,J=16.0 Hz,1H),8.18(d,J=8.0 Hz,1H),8.01(d,J=16.0 Hz,1H),7.73(s,1H),7.71(s,1H),7.64(d,J=4.0 Hz,2H),7.45-7.42(m,1H),7.40(s,1H),7.38(s,1H),7.37-7.35(m,1H),7.32(t,J=8.0 Hz,2H),7.02(s,1H),7.00(s,1H),6.95(t,J=8.0 Hz, 1H), 4.18 (t, J = 4.0 Hz, 2H), 4.13 (t, J = 4.0 Hz, 2H), 3.43 (t, J = 4.0 Hz, 4H); 13 C NMR(100 MHz,Chloroform-d)δ191.10,167.54,151.31,144.48,142.02,141.55,136.63,135.17,133.61,132.58,130.93,129.85,129.44,129.21 ,128.89,122.35,121.54,121.30,121.11,120.71,120.69,116.81,112.39,50.53,49.93,47.83,43.18; HRMS(ESI-MS)m / z:[M+H]+calcd for C 31 H 26 N4O2+:487.2136; found:487.2094.

[0084] Example 2

[0085] This example provides a phenylpiperazine β-carboline derivative I-2, whose structural formula is as follows:

[0086]

[0087] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0088] Its NMR and mass spectrometry data are as follows:

[0089] Yellow sold; yield 66%; mp341.9-343.0℃; 1H NMR (400 MHz, DMSO-d6) δ12.27(s,1H),8.87(s,1H),8.44-8.38(m,2H),7.95(d,J=12.0 Hz,1H),7.92(s,1H),7.91(d,J=4.0 Hz,3H),7.88(d,J=8.0Hz,1H),7.65(t,J=8.0Hz,1H),7.36(t,J=8.0Hz,1H),7.25(t,J=8.0Hz,3H),7.22 (t,J=7.0Hz,4H),7.01(s,1H),6.99(s,1H),6.86(t,J=8.0Hz,1H),3.97(d,J=12.0Hz,4H),3.34(s,1H); 13 C NMR(100MHz,DMSO-d6)δ189.26,166.56,164.62,162.15,151.02,142.25,141 .72,141.33,134.97,133.45,131.96,131.42(d,J=3.0Hz),130.99(d,J=9.0H z),129.40,128.99,122.24,121.19,120.73(d,J=10.0Hz),120.23,119.48,1 16.18,115.97,113.33,49.45,48.65,46.98,42.21; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 FN4O2 + :505.2042;found:505.2000.

[0090] Example 3

[0091] This example provides a phenylpiperazine β-carboline derivative I-3, whose structural formula is as follows:

[0092]

[0093] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0094] Its NMR and mass spectrometry data are as follows:

[0095] Yellow sold; yield 64%; mp318.9-320.0℃; 1H NMR(500MHz,Chloroform-d)δ10.61(s,1H),8.78(s,1H),8.48(d,J=15.0Hz,1H),8.19(d,J=10.0 Hz,1H),7.95(d,J=15.0Hz,1H),7.64(s,2H),7.51(d,J=10.0Hz,1H),7.42-7.38(m,2H),7.37-7.3 4(m,1H),7.31(t,J=10.0Hz,2H),7.13(t,J=10.0Hz,1H),7.00(d,J=5.0Hz,2H),6.95(t,J=10.0H z,1H),4.17(t,J=5.0Hz,2H),4.13(t,J=5.0Hz,2H),3.43(t,J=5.0Hz,2H),3.39(t,J=5.0Hz,2H); 13 C NMR (125MHz, Chloroform-d) δ 190.91, 167.53, 164.19, 162.23, 151.24, 142.98 (d, J = 2.5Hz), 142. 14,141.54,137.43(d,J=7.5Hz),136.66,133.45,132.65,130.75(d,J=8.8Hz),129.95,129.45,12 4.50(d,J=2.5Hz),122.16(d,J=58.8Hz),121.52(d,J=30.0Hz),121.08,120.77,117.73(d,J=21. 3Hz), 116.86, 115.32 (d, J = 21.3Hz), 112.40, 50.40, 50.01, 47.79, 43.15; HRMS (ESI-MS) m / z: [M+H] + calcd for C 31 H 25 FN4O2 + :505.2042;found:505.2000.

[0096] Example 4

[0097] This example provides a phenylpiperazine β-carboline derivative I-4, whose structural formula is as follows:

[0098]

[0099] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0100] Its NMR and mass spectrometry data are as follows:

[0101] Yellow sold; yield 62%; mp368.0-369.3℃; 1 H NMR(400MHz,Chloroform-d)δ10.63(s,1H),8.79(s,1H),8.58(d,J=16.0Hz,1H),8.18 (d,J=8.0Hz,1H),8.12(d,J=16.0Hz,1H),7.72(t,J=8.0Hz,1H),7.64(d,J=4.0Hz,2H), 7.43-7.35(m,2H),7.31(t,J=8.0Hz,2H),7.16-7.10(m,2H),7.01(d,J=8.0Hz,2H),6.9 4(t,J=7.3Hz,1H), 4.20(t,J=4.0Hz,2H), 4.13(t,J=4.0Hz,2H), 3.42(q,J=4.0Hz,4H); 13 C NMR(100MHz,Chloroform-d)δ191.12,167.46,163.32,160.79,151.35,142.09,141.54,137.04( d,J=2.0Hz),136.66,133.47,132.60,132.23(d,J=8.0Hz),129.95(d,J=3.0Hz),129.87,129.39, 124.75(d,J=4.0Hz),123.38,123.24(d,J=6.0Hz),122.35,121.51(d,J=11.0Hz),,121.11,120. 61,116.80,121.51(d,J=11.2Hz),,112.39,50.32,49.95,47.78,43.20; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 FN4O2 + :505.2042;found:505.2002.

[0102] Example 5

[0103] This example provides a phenylpiperazine β-carboline derivative I-5, whose structural formula is as follows:

[0104]

[0105] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0106] Its NMR and mass spectrometry data are as follows:

[0107] Yellow sold; yield 60%; mp312.6-313.8℃; 1 H NMR(400MHz,DMSO-d6)δ12.30(s,1H),8.88(s,1H),8.46(d,J=8.0Hz,1H),8.43 (s,1H),7.92(d,J=16.0Hz,1H),7.87(dd,J=12.0,4.0Hz,3H),7.65(t,J=8.0Hz, 1H),7.44(d,J=8.0Hz,2H),7.36(t,J=8.0Hz,1H),7.25(t,J=8.0Hz,2H),7.00( s,1H),6.99(s,1H),6.86(t,J=8.0Hz,1H),3.97(d,J=16.0Hz,4H),3.34(s,4H); 13 C NMR (125MHz, DMSO-d6) δ189.25,166.53,151.05,142.28,141.45,141.33,135.15,134.98,133.73,133.39,132.01,130.31,129.44,12 9.12,128.98,122.28,122.10,120.82,120.79,120.24,119.52,116.03,113.36,49.48,48.68,47.00,42.25; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 ClN4O2 + :521.1747; found:521.1707.

[0108] Example 6

[0109] This example provides a phenylpiperazine β-carboline derivative I-6, whose structural formula is as follows:

[0110]

[0111] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0112] Its NMR and mass spectrometry data are as follows:

[0113] Yellow sold; yield 64%; mp296.1-297.3℃; 1 H NMR(400MHz,Chloroform-d)δ10.61(s,1H),8.76(s,1H),8.46(d,J=16.0Hz,1 H),8.17(d,J=8.0Hz,1H),7.90(d,J=16.0Hz,1H),7.66-7.63(m,3H),7.59(d, J=8.0Hz,1H),7.38(d,J=8.0Hz,2H),7.31(d,J=8.0Hz,2H),7.29(s,1H),7.00 (d,J=8.0Hz,2H),6.94(t,J=5.0Hz,1H),4.17-4.12(m,4H),3.43-3.38(m,4H); 13 C NMR(101MHz,Chloroform-d)δ190.81,167.58,151.26,142.70,142.15,141 .56,136.99,136.63,135.21,133.45,132.63,130.66,130.40,129.93,129 .48,129.43,128.95,126.48,122.36,121.98,121.62,121.35,121.07,120 .77,116.93,112.40,50.58,49.84,47.81,43.13; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 ClN4O2 + :521.1747; found:521.1706.

[0114] Example 7

[0115] This example provides a phenylpiperazine β-carboline derivative I-7, whose structural formula is as follows:

[0116]

[0117] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0118] Its NMR and mass spectrometry data are as follows:

[0119] Yellow sold; yield 63%; mp379.0-380.2℃; 1H NMR (500 MHz, Chloroform-d) δ10.64(s,1H),8.77(s,1H),8.50-8.41(m,2H),8.19(d,J=5.0 Hz,1H),7.86(dd,J=10.0,5.0 Hz,1H),7.67-7.64(m,2H),7.47(dd,J=5.0,1.2 Hz,1H),7.38(ddd,J=8.0,5.5,2.6Hz,1H),7.34(td,J=10.0,5.0 Hz,1H),7.30(td,J=10.0,5.0 Hz,2H),7.21(t,J=10.0Hz,1H),6.98(d,J=10.0 Hz,2H),6.94(t,J=10.0 Hz,1H),4.15-4.11(m,4H),3.41(t,J=5.0 Hz,2H),3.38(t,J=5.0 Hz,2H); 13 C NMR(125 MHz,Chloroform-d)δ190.81,167.55,151.27,142.07,141.55,140.07,136.67,136.01,133.47,133.37,132.63,131.57,130.52,129.93,129.42 ,127.88,127.36,123.01,122.36,121.61,121.33,121.06,120.72,116.81,112.41,50.51,49.88,47.81,43.12; HRMS(ESI-MS)m / z:[M+H]+calcd for C 31 H 25 ClN4O2+:521.1747; found:521.1706.

[0120] Example 8

[0121] This example provides a phenylpiperazine β-carboline derivative I-8, whose structural formula is as follows:

[0122]

[0123] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0124] Its NMR and mass spectrometry data are as follows:

[0125] Yellow sold; yield 65%; mp349.5-350.3℃; 1H NMR (500 MHz, Chloroform-d) δ10.61 (s, 1H), 8.74 (s, 1H), 8.47 (d, J = 15.0 Hz, 1H), 8.16 (d, J = 10.0 Hz, 1H), 7.90 (d, J = 15.0 Hz, 1H), 7.64 (d, J = 5.0 Hz,2H),7.54(d,J=10.0 Hz,2H),7.47(d,J=10.0 Hz,2H),7.39-7.36(m,1H),7.34(t,J=5.0 Hz,2H),6.99(d,J=10.0 Hz,3H),4.13(t,J=5.0Hz,4H),3.43-3.39(m,4H); 13 CNMR(125 MHz,Chloroform-d)δ190.84,167.54,151.29,142.89,142.05,141.53,136.59,134.05,133.45,132.61,132.46,130.13,129.91,129.49 ,125.22,122.36,121.60,121.32,121.28,121.05,120.89,116.82,112.39,50.71,49.84,47.84,43.15; HRMS(ESI-MS)m / z:[M+H]+calcd forC 31 H 25 BrN4O2+:565.1242; found:565.1202.

[0126] Example 9

[0127] This example provides a phenylpiperazine β-carboline derivative I-9, whose structural formula is as follows:

[0128]

[0129] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0130] Its NMR and mass spectrometry data are as follows:

[0131] Yellow sold; yield 61%; mp330.8-332.1℃; 1H NMR(400MHz,Chloroform-d)δ10.49(s,1H),8.65(s,1H),8.34(d,J=16.0Hz,1H),8.06(d,J=8.0Hz,1H),7.77(d,J=16.0Hz,1H),7.70(s,1H),7.52(s,3H) ,7.41(d,J=8.0Hz,1H),7.18(t,J=8.0Hz,2H),7.14(s,1H),7.10(t,J=8.0Hz ,1H),6.88(d,J=8.0Hz,2H),6.82(t,J=8.0Hz,1H),4.02(s,4H),3.28(s,4H); 13 C NMR(100MHz,Chloroform-d)δ190.79,167.62,151.28,142.60,142.17,141.56,137.27,136.64,133.57,133.46,132.65,131.94,130.65,129.94 ,129.44,126.87,123.32,122.38,122.01,121.63,121.35,121.08,120. 77,116.97,112.40,50.65,49.78,47.83,43.12; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 BrN4O2 + :565.1242; found:565.1200.

[0132] Example 10

[0133] This example provides a phenylpiperazine β-carboline derivative I-10, whose structural formula is as follows:

[0134]

[0135] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0136] Its NMR and mass spectrometry data are as follows:

[0137] Yellow sold; yield 61%; mp328.5-329.4℃; 1H NMR(500MHz,Chloroform-d)δ10.64(s,1H),8.77(s,1H),8.46-8.37(m,2H),8.19(d,J=5.0 Hz,1H),7.86-7.84(m,1H),7.67(d,J=2.4Hz,1H),7.64(d,J=5.0Hz,2H),7.38(td,J=5.8,2. 9Hz,1H),7.32-7.28(m,2H),7.28-7.26(m,1H),7.26-7.24(m,1H),6.97(d,J=10.0Hz,2H), 6.94(t,J=5.0Hz,1H), 4.13(t,J=10.0Hz,4H), 3.41(t,J=5.0Hz,2H), 3.37(t,J=4.0Hz,2H); 13 C NMR(125MHz,Chloroform-d)δ190.74,167.55,151.27,142.71,142.07,141.56,136.67,135.11,133.80,133.45,132.64,131.73,129.94,129.42 ,128.02,128.00,126.54,123.21,122.37,121.62,121.33,121.06,120. 72,116.81,112.41,50.51,49.87,47.81,43.12; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 BrN4O2 + :565.1242; found:565.1196.

[0138] Example 11

[0139] This example provides a phenylpiperazine β-carboline derivative I-11, whose structural formula is as follows:

[0140]

[0141] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0142] Its NMR and mass spectrometry data are as follows:

[0143] Yellow sold; yield 61%; mp357.6-358.9℃; 1H NMR(500MHz,Chloroform-d)δ10.66(s,1H),8.77(s,1H),8.46(d,J=20.0Hz,1H),8.17(d,J= 10.0Hz,1H),7.98(d,J=20.0Hz,1H),7.63(d,J=5.0Hz,2H),7.61(d,J=5.0Hz,2H),7.38-7.3 5(m,1H),7.32(t,J=10.0Hz,2H),7.17(d,J=5.0Hz,2H),7.02(d,J=5.0Hz,2H),6.96(t,J=10 .0Hz,1H),4.18(t,J=5.0Hz,2H),4.13(t,J=5.0Hz,2H),3.43(t,J=5.0Hz,4H),2.40(s,3H); 13 C NMR(125MHz,Chloroform-d)δ191.13,167.56,151.34,144.53,141.92,141.54,141.52,136.59,133.70,132.51,132.45,129.96,129.79,12 9.42,128.91,122.32,121.48,121.22,121.09,120.68,119.71,116.84,112.37,50.53,49.92,47.84,43.18,21.75; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O2 + :501.2293;found:501.2253.

[0144] Example 12

[0145] This example provides a phenylpiperazine β-carboline derivative I-12, whose structural formula is as follows:

[0146]

[0147] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0148] Its NMR and mass spectrometry data are as follows:

[0149] Yellow sold; yield 60%; mp389.4-390.7℃; 1H NMR(400MHz,Chloroform-d)δ10.69(s,1H),8.81(s,1H),8.51(d,J=16.0Hz,1H),8.22(d,J=8.0Hz, 1H),8.02(d,J=16.0Hz,1H),7.67(d,J=4.0Hz,2H),7.58(d,J=8.0Hz,1H),7.54(s,1H),7.43-7.39( m,1H),7.35(d,J=8.0Hz,1H),7.31(d,J=4.0Hz,1H),7.30-7.27(m,2H),7.03(d,J=8.0Hz,2H),6.97 (t,J=8.0Hz,1H),4.21(t,J=4.0Hz,2H),4.16(t,J=4.0Hz,2H),3.46(t,J=4.0Hz,4H),2.34(s,3H); 13 CNMR(100MHz,Chloroform-d)δ191.16,167.65,151.31,144.75,142.02,141 .55,138.94,136.63,135.12,133.71,132.55,131.81,129.83,129.78,129.4 3,129.07,125.84,122.34,121.52,121.23,121.11,120.73,120.48,116.87, 116.85,112.38,50.64,49.81,47.86,43.14,21.41; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O2 + :501.2293;found:501.2249.

[0150] Example 13

[0151] This example provides a phenylpiperazine β-carboline derivative I-13, whose structural formula is as follows:

[0152]

[0153] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0154] Its NMR and mass spectrometry data are as follows:

[0155] Yellow sold; yield 61%; mp369.1-370.3℃;1 H NMR(500MHz,Chloroform-d)δ10.66(s,1H),8.79(s,1H),8.45(d,J=15.0Hz,1H),8.34(d,J= 15.0Hz,1H),8.20(d,J=5.0Hz,1H),7.83(d,J=5.0Hz,1H),7.65-7.64(m,2H),7.40-7.37(m,1 H),7.35-7.29(m,3H),7.26(d,J=10.0Hz,1H),7.18(t,J=5.0Hz,1H),6.99(d,J=5.0Hz,2H),6 .95(t,J=5.0Hz,1H),4.18(t,J=5.0Hz,2H),4.12(t,J=5.0Hz,2H),3.41(m,4H),2.56(s,3H); 13 CNMR(125MHz,Chloroform-d)δ191.18,167.51,151.29,142.03,141.97,14 1.53,138.84,136.69,134.02,133.65,132.58,131.20,130.73,129.85,12 9.42,126.68,126.63,122.37,121.55,121.51,121.33,121.11,120.70,11 6.84,112.36,50.43,49.99,47.82,43.18,20.03; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O2 + :501.2293;found:501.2250.

[0156] Example 14

[0157] This example provides a phenylpiperazine β-carboline derivative I-14, whose structural formula is as follows:

[0158]

[0159] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0160] Its NMR and mass spectrometry data are as follows:

[0161] Yellow sold; yield 64%; mp315.8-216.9℃; 1H NMR (400MHz, DMSO-d6) δ12.27(s,1H),8.86(s,1H),8.42(d,J=8.0Hz,1H),8.33(d ,J=16.0Hz,1H),7.92-7.86(m,2H),7.78(d,J=8.0Hz,2H),7.64(t,J=8.0Hz,1H), 7.34(t,J=8.0Hz,1H),7.26(t,J=8.0Hz,2H),7.02(d,J=8.0Hz,2H),6.92(d,J=8. 0Hz, 2H), 6.86 (t, J = 8.0Hz, 1H), 3.98 (d, J = 20.0Hz, 4H), 3.81 (s, 3H), 3.40 (s, 4H); 13 C NMR(100MHz,DMSO-d6)δ189.23,166.64,161.49,151.09,143.01,142.25,141.29,134.97,133.70,131.92,130.58,129.40, 129.05,127.37,122.25,120.78,120.56,120.26,119.55,118.69,116.05,114.59,113.35,55.44; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O3 + :517.2243; found:517.2202.

[0162] Example 15

[0163] This example provides a phenylpiperazine β-carboline derivative I-15, whose structural formula is as follows:

[0164]

[0165] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0166] Its NMR and mass spectrometry data are as follows:

[0167] Yellow sold; yield 63%; mp331.3-332.2℃; 1H NMR(400MHz,Chloroform-d)δ10.56(s,1H),8.70(s,1H),8.39(d,J=16.0Hz,1H),8.10(d,J=8.0H z,1H),7.89(d,J=16.0Hz,1H),7.55(d,J=4.0Hz,2H),7.30(dd,J=8.0,4.0Hz,1H),7.24(d,J=4.0H z,1H),7.23-7.21(m,2H),7.20-7.18(m,1H),7.13(s,1H),6.92(s,1H),6.90-6.88(m,2H),6.85( t,J=8.0Hz,1H),4.10(t,J=4.0Hz,2H),4.04(t,J=4.0Hz,2H),3.68(s,3H),3.33(t,J=4.0Hz,4H); 13 C NMR(100MHz,Chloroform-d)δ191.10,167.62,160.15,151.24,144.46,142.05,141.55,136.64,136.54,133.63,132.58,130.17,129.86,129.43,12 2.35,121.55,121.30,121.21,121.11,120.98,120.67,116.80,116.70,11 4.10,112.38,55.37,50.50,49.79,47.84,43.13; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O3 + :517.2243; found:517.2200.

[0168] Example 16

[0169] This example provides a phenylpiperazine β-carboline derivative I-16, whose structural formula is as follows:

[0170]

[0171] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0172] Its NMR and mass spectrometry data are as follows:

[0173] Yellow sold; yield 61%; mp347.5-348.4℃; 1H NMR(400MHz,Chloroform-d)δ10.69(s,1H),8.78(s,1H),8.54-8.38(m,2H),8.19(d,J= 8.0Hz,1H),7.76(d,J=8.0Hz,1H),7.63(d,J=4.0Hz,2H),7.40(t,J=8.0Hz,1H),7.38-7 .35(m,1H),7.30(t,J=8.0Hz,1H),6.99(s,1H),6.97(s,1H),6.96(d,J=4.0Hz,1H),6.9 4-6.91(m,2H),4.18(t,J=4.0Hz,2H),4.12(t,J=4.0Hz,2H),3.89(s,3H),3.41(m,4H); 13 C NMR(100MHz,Chloroform-d)δ191.52,167.65,159.10,151.30,141.95,141.53,139.78,136.65,133.90,132.47,132.27,129.75,129.41,12 8.89,124.22,122.32,121.44,121.13,121.01,120.59,116.74,112.37,111.50,55.79,50.38,49.89,47.85,43.17; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O3 + :517.2243; found:517.2200.

[0174] Example 17

[0175] This example provides a phenylpiperazine β-carboline derivative I-17, whose structural formula is as follows:

[0176]

[0177] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0178] Its NMR and mass spectrometry data are as follows:

[0179] Yellow sold; yield 64%; mp363.7-364.9℃; 1H NMR (400 MHz, Chloroform-d) δ10.60(s,1H),8.75(s,1H),8.54(d,J=16.0 Hz,1H),8.16(d,J=8.0 Hz,1H),7.96(d,J=16.0 Hz,1H),7.78(s,1H),7.76(s,1H),7.64(d,J=4.0 Hz,2H),7.59(s,1H),7.57(s,1H),7.40-7.36(m,1H),7.32(t,J=8.0 Hz,2H),7.00-6.98(m,2H),6.97(d,J=8.0 Hz,1H),4.13(s,4H),3.42(t,J=4.0Hz,4H); 13 C NMR(100 MHz,Chloroform-d)δ190.69,167.51,151.31,142.17(d,J=3.0 Hz),141.57,138.48,136.63,133.30,132.70,129.99,129.48,128.84,126.18(d,J=4.0 Hz),126.10(d,J=4.0 Hz),123.02,122.39,121.68,121.40,121.05,120.96,116.84,112.42,50.89,49.80,47.86,43.17; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 25 F3N4O2 + :555.2011; found:555.1963. Example 18

[0180] This example provides a phenylpiperazine β-carboline derivative I-18, whose structural formula is as follows:

[0181]

[0182] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0183] Its NMR and mass spectrometry data are as follows:

[0184] Yellow sold; yield 62%; mp316.4-317.7℃; 1H NMR (500 MHz, Chloroform-d) δ10.61(s,1H),8.76(s,1H),8.53(d,J=15.0 Hz,1H),8.19(d,J=5.0 Hz,1H),7.99(d,J=15.0 Hz,1H),7.93(d,J=5.0 Hz,1H),7.90(s,1H),7.66(d,J=15.0 Hz,3H),7.47(t,J=10.0 Hz,1H),7.40-7.37(m,1H),7.30(t,J=10.0Hz,2H),6.98(d,J=10.0Hz,2H),6.94(t, J=10.0Hz,1H),4.13(t,J=5.0Hz,4H),3.42(t,J=5.0Hz,2H),3.36(t,J=5.0Hz,2H); 13 CNMR(125MHz,Chloroform-d)δ190.75,167.66,151.22,142.45,142.19,141.56,136 .64,135.95,133.43,132.68,131.73(d,J=32.5Hz),130.85,129.99,129.71,129.43 ,127.18(d,J=3.8Hz),126.18(d,J=3.8Hz),122.39(d,J=2.1Hz),121.67,121.32,12 1.05,120.77,116.86,112.41,50.61,49.76,47.81,43.08; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 25 F3N4O2 + :555.2011; found:555.1964. Example 19

[0185] This example provides a phenylpiperazine β-carboline derivative I-19, whose structural formula is as follows:

[0186]

[0187] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0188] Its NMR and mass spectrometry data are as follows:

[0189] Yellow sold; yield 59%; mp339.0-340.1℃; 1H NMR(400MHz,Chloroform-d)δ10.63(s,1H),8.77(s,1H),8.47(d,J=16.0Hz,1H),8.38(d ,J=16.0Hz,1H),8.19(d,J=8.0Hz,1H),7.95(d,J=8.0Hz,1H),7.76(d,J=8.0Hz,1H),7.68 -7.62(m 2H),7.50(m,2H),7.39(t,J=8.0Hz,1H),7.30(t,J=8.0Hz,2H),6.94(t,J=8. 0Hz, 3H), 4.11 (t, J = 4.0Hz, 4H), 3.40 (t, J = 5.0Hz, 2H), 3.35 (t, J = 5.0Hz, 2H); 13 C NMR(100MHz,Chloroform-d)δ190.56,167.58,151.28,142.14,141.60,139.5 6,136.71,134.11,134.09,133.39,132.70,132.36,130.02(d,J=5.0Hz),129 .42,128.06,126.50(d,J=5.0Hz),124.54,122.38,121.67,121.32,121.06,1 20.74,116.81,112.42,50.56,49.83,47.81,43.10; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 25 F3N4O2 + :555.2011;found:555.1963.

[0190] Example 20

[0191] This example provides a phenylpiperazine β-carboline derivative I-20, whose structural formula is as follows:

[0192]

[0193] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0194] Its NMR and mass spectrometry data are as follows:

[0195] Yellow sold; yield 62%; mp378.4-379.4℃; 1H NMR(500MHz,Chloroform-d)δ10.57(s,1H),8.76(s,1H),8.58(d,J=20.0Hz,1H),8.21(d,J=10.0Hz,1H),7.95(d,J=20.0Hz,1H),7.78(d,J=5.0Hz,2H) ,7.67(m,2H),7.62(d,J=10.0Hz,2H),7.41(m,1H),7.33(t,J=10.0Hz,2H), 6.99(t,J=10.0Hz,3H),4.12(s,2H),4.08(s,2H),3.41(s,2H),3.38(s,2H); 13 C NMR(150MHz,DMSO-d6)δ189.22,166.50,151.01,142.29,141.40,140.62,139.32,134.99,133.26,132.81,132.07,129.48,129.18,126.8 4,124.76,122.29,120.87,120.22,119.48,118.52,115.99,113.35,112.31,79.17,49.45,48.61,46.96,42.20; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 25 N5O2 + :512.2089; found:512.2044.

[0196] Example 21

[0197] This example provides a phenylpiperazine β-carboline derivative I-21, whose structural formula is as follows:

[0198]

[0199] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0200] Its NMR and mass spectrometry data are as follows:

[0201] Yellow sold; yield 61%; mp316.0-317.2℃; 1H NMR(500MHz,Chloroform-d)δ10.60(s,1H),8.73(s,1H),8.51(d,J=15.0Hz,1H),8.18(d,J=10.0Hz ,1H),7.96(d,J=10.0Hz,1H),7.91(d,J=20.0Hz,2H),7.68(d,J=5.0Hz,1H),7.66(d,J=5.0Hz,2H), 7.44(t,J=10.0Hz,1H),7.41-7.37(m,1H),7.32-7.29(m,2H),6.98(d,J=5.0Hz,2H),6.95(t,J=5.0 Hz,1H),4.13(t,J=5.0Hz,2H),4.08(t,J=5.0Hz,2H),3.41(t,J=5.0Hz,2H),3.33(t,J=5.0Hz,2H); 13 C NMR(125MHz,Chloroform-d)δ190.56,167.65,151.20,142.24,141.57,141 .37,136.62,136.41,133.69,133.36,132.97,132.70,131.48,130.04,130 .00,129.47,123.06,122.38,121.71,121.20,120.99,120.82,118.26,116 .82,113.63,112.44,50.62,49.73,47.78,43.01; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 25 N5O2 + :512.2089; found:512.2047.

[0202] Example 22

[0203] This example provides a phenylpiperazine β-carboline derivative I-22, whose structural formula is as follows:

[0204]

[0205] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0206] Its NMR and mass spectrometry data are as follows:

[0207] Orange sold; yield 64%; mp361.3-362.5℃; 1H NMR (600MHz, DMSO-d6) δ12.34(s,1H),8.90(s,1H),8.57(d,J=12.0Hz,1H),8.44(d,J=6.0Hz, 1H),8.20(d,J=12.0Hz,2H),8.09(d,J=6.0Hz,2H),8.00(d,J=12.0Hz,1H),7.88(d,J=6.0Hz, 1H),7.66(t,J=6.0Hz,1H),7.36(t,J=6.0Hz,1H),7.24(t,J=6.0Hz,2H),6.99(d,J=12.0Hz,2 H),6.85(t,J=6.0Hz,1H),3.99(d,J=6.0Hz,2H),3.95(t,J=6.0Hz,2H),3.34(d,J=6.0Hz,4H); 13 C NMR(150MHz,DMSO-d6)δ189.12,166.47,151.02,148.03,142.30,141.40,141.19,139.99,135.02,133.19,132.10,129.58,129.50,12 8.94,125.39,124.07,122.30,120.97,120.89,120.23,119.50,115.96,113.36,49.42,48.65,46.96,42.25; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 N5O4 + :532.1987; found:532.1945.

[0208] Example 23

[0209] This example provides a phenylpiperazine-based β-carboline derivative I-23, whose structural formula is as follows:

[0210]

[0211] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0212] Its NMR and mass spectrometry data are as follows:

[0213] Orange sold; yield 62%; mp325.6-326.6℃; 1H NMR(500MHz,Chloroform-d)δ10.58(s,1H),8.77s,1H),8.60(d,J=20.0Hz,1H),8.53(t,J=5 .0Hz,1H),8.27(dd,J=10.0,5.0Hz,1H),8.21(d,J=5.0Hz,1H),8.05(d,J=10.0Hz,1H),8.02( d,J=20.0Hz,1H),7.69-7.65(m,2H),7.54(t,J=10.0Hz,1H),7.42-7.39(m,1H),7.31–7.28(m ,2H),6.97(d,J=10.0Hz,2H),6.94(t,J=10.0Hz,1H),4.12(s,4H),3.42(s,2H),3.36(s,2H); 13 C NMR(100MHz,Chloroform-d)δ190.51,167.64,151.19,148.87,142.27,141.59,141.21,136.94,136.63,133.46,133.32,132.74,130.16,130.05 ,129.42,124.94,123.76,123.60,122.38,121.73,121.30,121.02,120. 79,116.83,112.45,50.64,49.69,47.82,43.07; HRMS(ESI-MS)m / z:[M+H] + calcd for C 31 H 25 N5O4 + :532.1987; found:532.1945.

[0214] Example 24

[0215] This example provides a phenylpiperazine β-carboline derivative I-24, whose structural formula is as follows:

[0216]

[0217] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0218] Its NMR and mass spectrometry data are as follows:

[0219] Orange sold; yield 60%; mp347.8-348.9℃; 1H NMR (600MHz, DMSO-d6) δ12.34(s,1H),8.91(s,1H),8.57(d,J=18.0Hz,1H),8.44(d,J=6.0Hz,1H) ,8.12(s,1H),8.10(s,1H),8.00(d,J=18.0Hz,1H),7.96(s,1H),7.95(s,1H),7.88(d,J=12.0Hz, 1H),7.66(t,J=12.0Hz,1H),7.37(t,J=6.0Hz,1H),7.26(t,J=6.0Hz,2H),7.01(s,1H),7.00(s,1 H),6.85(t,J=6.0Hz,1H),4.01(t,J=6.0Hz,2H),3.95(t,J=6.0Hz,2H),3.36(m,4H),3.26(s,3H); 13 C NMR(150MHz,DMSO-d6)δ189.16,166.46,151.01,142.30,141.86,141.40,140.70,139.61,135.04,133.23,132.08,129.49,129.27,129.0 2,127.64,124.42,122.31,120.97,120.88,120.23,119.51,115.95,113.37,49.33,48.73,46.94,43.43,42.25; HRMS(ESI-MS)m / z:[M+H] + calcd for C 32 H 28 N4O4S + :565.1913; found:565.1830.

[0220] Example 25

[0221] This example provides a phenylpiperazine β-carboline derivative I-25, whose structural formula is as follows:

[0222]

[0223] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0224] Its NMR and mass spectrometry data are as follows:

[0225] Yellow sold; yield 65%; mp328.0-329.2℃; 1H NMR (400MHz, Chloroform-d) δ10.67(s,1H),8.79(s,1H),8.49(d,J=16.0Hz,1H),8.19(d,J=8. 0Hz,1H),8.00(d,J=16.0Hz,1H),7.68(s,1H),7.66(s,1H),7.64(d,J=4.0Hz,2H),7.40(s,1H) ,7.38(s,1H),7.34(t,J=8.0Hz,3H),7.05(s,1H),7.03(s,1H),6.97(t,J=8.0Hz,1H),4.20(t, J=4.0Hz,2H),4.13(t,J=4.0Hz,2H),3.49(t,J=4.0Hz,2H),3.44(t,J=4.0Hz,2H),1.35(s,9H); 13 C NMR(100MHz,Chloroform-d)δ191.15,167.57,154.72,151.39,144.44,141.96,141.55,136.64,133.71,132.56,132.42,129.81,129.45,128.8 0,126.22,122.34,121.50,121.29,121.13,120.72,119.78,116.88,112 .38,50.76,49.85,47.89,43.22,35.16,31.31; HRMS(ESI-MS)m / z:[M+H] + calcd for C 35 H 34 N4O2 + :543.2764; found:543.2719.

[0226] Example 26

[0227] This example provides a phenylpiperazine-based β-carboline derivative I-26, whose structural formula is as follows:

[0228]

[0229] The compound S5 was prepared by the general preparation method described above, and the structural formula thereof is:

[0230] Its NMR and mass spectrometry data are as follows:

[0231] Yellow sold; yield 62%; mp361.7-362.7℃; 1H NMR (400MHz, Chloroform-d) δ10.67(s,1H),8.74(s,1H),8.37(d,J=16.0Hz,1H),8.19(d,J=8.0Hz,1H),7.93(d,J=16.0Hz,1H),7.64(d,J= 4.0Hz,2H),7.40-7.36(m,1H),7.29(d,J=8.0Hz,2H),6.95(s,1H),6.94-6.90(m,4H),4.09(s,4H),3.89(s,3H),3.75(s,6H),3.39(s,4H); 13 C NMR(100MHz,Chloroform-d)δ190.93,168.02,153.63,151.05,144.74,142.06,141.56,140.83,136.63,133.81,132.58,130.62,129.88,129.5 0,122.34,121.55,121.09,121.03,120.83,119.82,116.70,112.38,106 .03,61.15,56.14,50.89,49.29,47.91,42.93; HRMS(ESI-MS)m / z:[M+H] + calcd for C 34 H 32 N4O5 + :577.2455; found:577.2406.

[0232] Performance testing

[0233] α-Glucosidase inhibition activity test

[0234] 1. Preparation of reagents and standard solutions

[0235] (1) 100 mM phosphate buffer (PBS, pH = 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve the dilution reagent.

[0236] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to a working concentration of 0.05 U / mL, and freeze in aliquots.

[0237] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex mix thoroughly and prepare it fresh before each experiment.

[0238] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0239] 2. Experimental steps

[0240] (1) 10 μL of α-glucosidase with a working concentration of 0.05 U / mL, 130 μL of 100 mM phosphate buffer (pH = 6.8), and 10 μL of different concentrations of compounds (phenylpiperazine-based β-carboline derivatives prepared in Examples 1 to 26) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of DMSO with a content equal to 5% by weight. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0241] (2) Subsequently, 50 μL of substrate PNPG was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the time was evenly distributed three times. The readings were read once at a wavelength of 405 nm during each time period. The readings were recorded as OD1, OD2, and OD3.

[0242] (3) The α-glucosidase inhibitory activity of the test compound was calculated according to the following formula:

[0243] Inhibition rate (%) = [(OD3-OD)-(OD1-OD)] / OD3-OD×100%

[0244] Where OD represents the absorbance value of the blank control group. Data processing: MS Excel was used to analyze and process the data, and Origin9.1 was used to calculate the half-maximal inhibitory concentration (IC 50 ), IC 50 It represents the concentration of the test compound required to inhibit the activity of α-glucosidase by 50% under the experimental conditions.

[0245] 3. Results Analysis

[0246] The α-glucosidase inhibitory activity of the synthesized compounds was evaluated by in vitro enzymatic assay, and the results are shown in Table 1:

[0247] Table 1 Evaluation of the in vitro α-glucosidase inhibitory activity of compounds I-1 to I-26

[0248]

[0249] The IC value of the positive control drug acarbose 50All the derivatives I-1 to I-26 prepared in Examples 1 to 26 have good α-glucosidase inhibitory activity, IC 50 The values ​​ranged from 4.85±0.48μM to 10.84±1.02μM, which were better than the positive control acarbose. 50 =4.85±0.48μM) had the most significant inhibitory effect, and its half-inhibitory concentration is shown in the figure. Figure 1 The results showed that these compounds showed strong binding affinity when interacting with α-glucosidase. 50 =180±3.20μM), the introduction of piperazine and chalcone structures can enhance its inhibitory activity against α-glucosidase.

[0250] The structural formula of β-carboline is:

[0251] Enzyme kinetics test

[0252] The α-glucosidase inhibitory activity of the synthesized compounds was evaluated using in vitro enzyme kinetics experiments:

[0253] 1. Preparation of reagents and standard solutions

[0254] 1) 100 mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and sodium dihydrogen phosphate and dissolve them in ultrapure water to dissolve the dilution reagent.

[0255] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to prepare working concentrations of 0.0375 U / mL, 0.05 U / mL, 0.0625 U / mL, and 0.075 U / mL, respectively, and freeze in aliquots.

[0256] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex mix thoroughly and prepare it fresh before each experiment.

[0257] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0258] 2. Experimental steps

[0259] (1) 10 μL of α-glucosidase at concentrations of 0.0375 U / mL, 0.05 U / mL, 0.0625 U / mL, and 0.075 U / mL, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compounds at different concentrations (Compound I-25 prepared in Example 25) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of 5% DMSO to replace 10 μL of the compound. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0260] (2) Subsequently, 50 μL of 0.25 mM PNPG was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the sample was read three times at a wavelength of 405 nm. The readings were recorded as OD1, OD2, and OD3.

[0261] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0262] 3. Results Analysis

[0263] The results of enzyme kinetic inhibition type evaluation test are as follows Figure 2 As shown. Figure 2 It can be seen that the inhibitory effect is reversible.

[0264] Substrate kinetics experiments

[0265] In vitro substrate kinetics experiments were used to evaluate the α-glucosidase inhibitory activity of the synthesized active compounds:

[0266] 1. Preparation of reagents and standard solutions

[0267] (1) 100 mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve the dilution reagent.

[0268] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100 mM PBS to an enzyme with an activity of 100 U to a working concentration of 0.05 U / mL, and freeze in aliquots.

[0269] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoglucoside (PNPG), dissolve it in 100 mM PBS solution, and prepare substrate working solutions with concentrations of (0.25 mM, 0.5 mM, 0.75 mM, 1 mM). Vortex mix thoroughly and prepare freshly before each experiment.

[0270] Preparation of test drugs: Accurately weigh an appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20°C in the dark. Before the experiment, dilute it with DMSO to the desired concentration (0-200 μM), with a DMSO content of 5%.

[0271] 2. Experimental steps

[0272] (1) 10 μL of 0.5 U / mL α-glucosidase, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of different concentrations of compound (Compound I-25 prepared in Example 25) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of 5% DMSO to replace 10 μL of compound. Acarbose was used as a positive control. Four replicate wells were set up in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37°C for 10 min.

[0273] (2) Subsequently, 50 μL of substrate PNPG of different concentrations was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the time was evenly distributed three times, and the reading was taken once at a wavelength of 405 nm during each time period. The readings were recorded as OD1, OD2, and OD3.

[0274] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0275] 3. Results Analysis

[0276] The results of the substrate kinetic inhibition type evaluation experiment are as follows Figure 3 As shown, Figure 3 The corresponding chalcone-containing β-carboline derivative I-25 has a substrate kinetics diagram for α-glucosidase in vitro. Figure 3 It can be seen that all the straight lines almost intersect at the x-axis, and the Michaelis-Menten constant (Km) remains unchanged, demonstrating that compound I-25 has the effect of a non-competitive inhibitor.

[0277] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the scope of the present invention.

Claims

1. A phenylpiperazine-β-carboline derivative, characterized in that: It has the structural formula shown in formula (I): Wherein, R is one or more substituents, or R does not exist; R is independently selected from F, Cl, Br, C 1～4 Alkyl, C 1～3 Alkoxy, C 1～3 of haloalkyl, cyano, nitro, and methylsulfonyl.

2. The phenylpiperazine-based β-carboline derivative according to claim 1, characterized in that: Selected from one of the following structural formulas:

3. A method for preparing the phenylpiperazine-based β-carboline derivatives according to claim 1 or 2, characterized in that: The steps include: Compound S4, compound S5 and a base are mixed and subjected to a condensation reaction to obtain the phenylpiperazine β-carboline derivative; The structural formulas of compound S4 and compound S5 are shown below:

4. The preparation method according to claim 3, characterized in that The molar ratio of the compound S4 to the compound S5 is 1:(1-1.5).

5. The preparation method according to claim 3, characterized in that The alkali is selected from at least one of potassium hydroxide and sodium hydroxide.

6. A pharmaceutical composition, characterized in that The invention comprises the phenylpiperazine-β-carboline derivative according to claim 1 or 2; and pharmaceutically acceptable excipients.

7. The pharmaceutical composition according to claim 6, characterized in that The dosage form of the pharmaceutical composition is selected from tablets, capsules, oral solutions or injections.

8. An α-glucosidase inhibitor, characterized in that The invention comprises the phenylpiperazine-based β-carboline derivatives according to claim 1 or 2.

9. Use of the phenylpiperazine-based β-carboline derivatives according to claim 1 or 2, the pharmaceutical composition according to claim 6 or 7, or the α-glucosidase inhibitor according to claim 8 in the preparation of a drug for treating and / or preventing diabetes.

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