Phenyl piperazinyl beta-carboline derivative as well as preparation method and application thereof

Abstract

The invention discloses a phenyl piperazinyl beta-carboline derivative as well as a preparation method and application thereof. The phenyl piperazinyl beta-carboline derivative has a structural formula as shown in a formula (I): # imgabs0 #, in which R is one or more substituent groups, or R does not exist; r is independently selected from halogen, a C1-6 alkyl group, a C1-6 alkoxy group, a C1-6 halogenated alkyl group, a cyano group, a nitro group and a C1-6 alkylsulfonyl group. A series of phenyl piperazinyl beta-carboline derivatives are designed and synthesized by taking a beta-carboline structure as a mother nucleus, the derivatives show a relatively strong alpha-glucosidase inhibition effect, and the IC50 value range of the derivatives is 4.85 + / -0.48 [mu] M to 10.84 + / -1.02. The compound can be used as an alpha-glucosidase inhibitor for treating and / or preventing diabetes mellitus.

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 and a preparation method and application thereof. Background Art

[0002] At present, diabetes has become a global problem, seriously endangering human health. Although people's health awareness has improved, the incidence of diabetes is still rising. If blood sugar levels are not effectively controlled in time, it will induce a series of functional disorders and damages in the human body, seriously threatening the health and safety of patients. Among them, type 2 diabetes is the most common, accounting for more than 90% of the number of patients. Type 2 diabetes, also known as non-insulin-dependent diabetes mellitus (NIDDM), is a chronic metabolic disease caused by insulin dysfunction or insulin resistance, mainly caused by the interaction of genetic and environmental factors, such as gene mutations, eating habits, etc.

[0003] There are many oral hypoglycemic drugs on the market, which act on different targets through different pathways. Sulfonylurea drugs (glibenclamide, glimepiride and mesitylamide) 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 hepatic bile 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. At present, 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, the first aspect of 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 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 is absent; R is independently selected from halogen, C 1～6 Alkyl, C 1～6 Alkoxy, C 1～6 haloalkyl, 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 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. 50 The value ranges 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 halogenated alkyl, cyano, nitro, 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 according to the first aspect of the present invention, comprising the following steps:

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

[0024] The structural formulas of compound S4 and compound S5 are as follows:

[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, 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, after the condensation reaction is completed, a purification step is further included, 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) urea 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, after the reaction is completed, a post-treatment step is further included; the post-treatment includes: 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 is used as a 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, 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 a 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 the 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] The fifth aspect of 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 1 to 6 carbon atoms, including C 1-6 Straight chain alkyl, C 1-6 The 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, for example, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a tert-butyl group, a n-pentyl group, an isopentyl group, a n-hexyl group, a cyclopropyl group, a methylcyclopropyl group, an ethylcyclopropyl group, a cyclopentyl group, a methylcyclopentyl group, a cyclohexyl group, and the like.

[0059] “C 1-6 "alkoxy" means an alkoxy group having 1 to 6 carbon atoms, including C 1-6 Straight chain alkoxy, C 3-6 Branched alkoxy and C 2-6The cycloalkoxy group may 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, such as methoxy, ethoxy, n-propoxy, isopropoxy, etc.

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

[0061] “C 1～6 The halogenated alkyl" and "C 1-6 The definition of "alkyl" is similar to that of "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 apparent 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 easily understood from the description of the embodiments in conjunction with the following drawings, in which:

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

[0066] Figure 2 The enzymatic kinetic diagram of compound I-25 prepared in Example 25 of the present invention as an α-glucosidase inhibitor on α-glucosidase in vitro;

[0067] Figure 3 This is a kinetic diagram of the substrate of α-glucosidase in vitro for compound I-25 prepared in Example 25 of the present invention as an α-glucosidase inhibitor. 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, acetone aldehyde 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 methyl acetone (9.6 mmol), and then slowly add 4 mL of concentrated sulfuric acid. React at room temperature for 24 hours and monitor with TLC plate spot plate (PE:EA-2:1). After the reaction is completed, adjust the pH to 7 with NaOH solution, let 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 and filter out the solid to obtain the product S3.

[0076] Synthesis of compound S4: 870 mg of compound S3 (3.42 mmol) was weighed into a 100 mL two-necked bottle, 1.3 g of HATU (3.42 mmol) was weighed therein, 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 the reaction was carried out at room temperature for 24 h. The reaction was monitored by TLC plate spot plate (PE:EA-2:1). After the reaction was completed, about 10 mL of water was added to quench the reaction, and the solid was filtered to obtain the 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, and add dropwise to a round-bottom flask after dissolution, 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, then recrystallize with 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 is prepared by the above general preparation method, 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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 is prepared by the above general preparation method, 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate for C 31 H 25 BrN4O2 + :565.1242; found:565.1196.

[0138] Embodiment 11

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

[0140]

[0141] The compound S5 is prepared by the above general preparation method, 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] + Calculate 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, the structural formula of which is as follows:

[0146]

[0147] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 28 N4O2 + :501.2293; found:501.2249.

[0150] Embodiment 13

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

[0152]

[0153] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 28 N4O2 + :501.2293; found:501.2250.

[0156] Embodiment 14

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

[0158]

[0159] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 28 N4O3 + :517.2243; found:517.2202.

[0162] Embodiment 15

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

[0164]

[0165] The compound S5 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate for C 32 H 28 N4O3 + :517.2243; found:517.2200.

[0174] Embodiment 17

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

[0176]

[0177] The compound S5 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate 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 is prepared by the above general preparation method, 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] + Calculate for C 32 H 25 F3N4O2 + :555.2011; found:555.1963.

[0190] Embodiment 20

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

[0192]

[0193] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 25 N5O2 + :512.2089; found:512.2044.

[0196] Embodiment 21

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

[0198]

[0199] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 25 N5O2 + :512.2089; found:512.2047.

[0202] Embodiment 22

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

[0204]

[0205] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 31 H 25 N5O4 + :532.1987; found:532.1945.

[0208] Embodiment 23

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

[0210]

[0211] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 31 H 25 N5O4 + :532.1987; found:532.1945.

[0214] Embodiment 24

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

[0216]

[0217] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 32 H 28 N4O4S + :565.1913; found:565.1830.

[0220] Embodiment 25

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

[0222]

[0223] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 35 H 34 N4O2 + :543.2764; found:543.2719.

[0226] Embodiment 26

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

[0228]

[0229] The compound S5 is prepared by the above general preparation method, 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] + Calculate for C 34 H 32 N4O5 + :577.2455; found:577.2406.

[0232] Performance Testing

[0233] α-Glucosidase Inhibitory 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 and dilute the 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), add 100 mM PBS solution to dissolve it, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex to mix well and prepare it fresh before each experiment.

[0238] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 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 compounds of different concentrations (phenylpiperazine 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%, and acarbose was used as a positive control. Four replicate wells were set in parallel for each group, and 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 incubation 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] 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 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 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 inhibition 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 improve its inhibitory activity against α-glucosidase.

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

[0251] Enzyme kinetics assay

[0252] The α-glucosidase inhibitory activity of the synthesized compounds was evaluated by 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 disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve and dilute the reagents.

[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 make 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), add 100 mM PBS solution to dissolve it, and prepare a substrate working solution with a concentration of 0.25 mM. Vortex to mix well and prepare it fresh before each experiment.

[0257] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 5%.

[0258] 2. Experimental steps

[0259] (1) 10 μL of α-glucosidase with 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 with 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 in parallel for each group, and 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 incubation time was evenly distributed three times. The readings were taken once at a wavelength of 405 nm during each time period. 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 △OD / min.

[0262] 3. Results Analysis

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

[0264] Substrate kinetics experiments

[0265] The in vitro substrate kinetics experiment was 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 sodium dihydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve and dilute the reagents.

[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), add 100 mM PBS solution to dissolve, and prepare substrate working solutions with concentrations of (0.25 mM, 0.5 mM, 0.75 mM, 1 mM). Vortex to mix well and prepare freshly before each experiment.

[0270] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 5%.

[0271] 2. Experimental steps

[0272] (1) 10 μL of 0.5 U / mL α-glucosidase, 130 μL (pH 6.8) 100 mM phosphate buffer, and 10 μL of different concentrations of compounds (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 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 readings were evenly distributed three times and read 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 △OD / 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 kinetic 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, proving 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 scope of ordinary technicians in the relevant technical field without departing from the purpose of the present invention.

Claims

1. A phenylpiperazine-based β-carboline derivative, characterized in that: It has the structural formula shown in formula (I): Wherein, R is one or more substituents, or R is absent; R is independently selected from halogen, C 1～6 Alkyl, C 1～6 Alkoxy, C 1～6 haloalkyl, cyano, nitro, C 1～6 of an alkylsulfonyl group.

2. The phenylpiperazine-based β-carboline derivative according to claim 1, characterized in that: R is one or more substituents, or R is absent; independently selected from halogen, C 1～4 Alkyl, C 1～3 Alkoxy, C 1～3 of halogenated alkyl, cyano, nitro, methylsulfonyl.

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

4. A method for preparing the phenylpiperazine-based β-carboline derivatives according to any one of claims 1 to 3, characterized in that: The steps include: Compound S4, compound S5 and a base are mixed for condensation reaction to obtain the phenylpiperazine β-carboline derivative; The structural formulas of compound S4 and compound S5 are as follows:

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

6. The preparation method according to claim 4, characterized in that: The alkali is selected from at least one of potassium hydroxide and sodium hydroxide.

7. A pharmaceutical composition, characterized in that The invention comprises the phenylpiperazine-based β-carboline derivative according to any one of claims 1 to 3; and pharmaceutically acceptable excipients.

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

9. An α-glucosidase inhibitor, characterized in that The invention comprises the phenylpiperazine-based β-carboline derivatives according to any one of claims 1 to 3.

10. Use of the phenylpiperazine-based β-carboline derivatives according to any one of claims 1 to 3, the pharmaceutical composition according to claim 7 or 8, or the α-glucosidase inhibitor according to claim 9 in the preparation of a drug for treating and / or preventing diabetes.

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