Nuciferine derivative as well as preparation method and application thereof

Abstract

The invention discloses a nuciferine derivative as well as a preparation method and application thereof, and relates to the technical field of medicines. The invention provides a nuciferine derivative and also provides a preparation method of the nuciferine derivative, the nuciferine derivative provided by the invention greatly improves the treatment effect of arrhythmia by inhibiting multiple ion channels, reduces the adverse reaction of drugs and reduces the risk of inducing ventricular arrhythmia, and the nuciferine derivative provided by the invention can be applied to treatment of ventricular arrhythmia. And the raw materials are low in price and easy to obtain, the production cost is greatly reduced, and the compound is suitable for large-scale production, has a certain treatment effect on the QT syndrome, and has a wide application prospect in preparation of medicines for resisting arrhythmia and treating the QT syndrome.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to nuciferine derivatives and preparation methods and applications thereof. Background Art

[0002] Arrhythmia refers to a cardiovascular disease caused by abnormalities in the frequency, rhythm, origin of heartbeats, direction and sequence of atrium and ventricle conduction. Common clinical symptoms include palpitations, and severe cases can lead to dizziness, blackouts, and even sudden death. Arrhythmia can exist alone or accompany other organic heart attacks. Its prognosis is closely related to the cause, inducement, and hemodynamics. The main pathogenesis of arrhythmia is that the origin and conduction function of cardiac activity are blocked, which leads to problems with the rhythm or frequency of heart beats. The occurrence of arrhythmia is usually accompanied by receptor blockage, changes in the current of myocardial cell ion channels, etc. Ideal antiarrhythmic drugs should act gently on the best targets, such as sodium ion channels and calcium ion channels.

[0003] There are two main ways to treat arrhythmias: one is drug therapy, and the other is pacemaker implantation. β-receptor agonists, M-cholinergic receptor blockers, phosphodiesterase inhibitors, etc. are all commonly used drugs in clinical practice. Isoproterenol, epinephrine, ephedrine, etc. are all β-adrenergic receptor agonists. M-cholinergic receptor blockers include atropine, etc., which can use cyclic adenosine monophosphate to enhance myocardial contractility and increase heart rate. In addition, aminophylline, glucocorticoids, thyroxine, etc. can work by specifically exciting the sinoatrial junction and improving atrioventricular conduction function. Although the use of Western medicine can significantly increase heart rate, it also has side effects such as dry mouth, facial flushing, dysuria, nausea and vomiting, irritability, insomnia, and rapid heart rate, and long-term use is generally not recommended. Pacemaker implantation is an effective strategy for the treatment of arrhythmias, but it is expensive and requires lifelong follow-up. The installation technology is limited and difficult to promote. There is also a certain risk of surgical trauma, which leads to accidents such as pacing failure and electrode dislocation.

[0004] In the field of TCM treatment, arrhythmia can be classified as "heart palpitations", and this disease is one of the dominant diseases in the field of TCM treatment. In the field of TCM, prescriptions used to treat arrhythmia are essentially natural compounds with safety, so TCM has a relatively greater advantage in treating arrhythmia.

[0005] Therefore, the development of a safe and highly effective antiarrhythmic drug has important scientific significance and economic value. Summary of the invention

[0006] The purpose of the present invention is to provide a nuciferine derivative and a preparation method and application thereof.

[0007] To achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0008] In one aspect, the present invention provides a nuciferine derivative, the structure of which is shown below:

[0009]

[0010] The R group structure is any of the following structures:

[0011]

[0012] In another aspect, the present invention provides a method for preparing the above-mentioned nuciferine derivatives, and the chemical reaction formula is as follows:

[0013]

[0014] The reaction conditions are: (a) EDCI, HOBt, DCM, rt, 24 hrs; (b) (i) PCl5, DCM, reflux, 20 mins; (ii) NaBH4, MeOH, 0℃ to rt, 2 hrs; (c) Et3N, TFAH, CH2Cl2, 1.5 hrs; (d) Pd(OAc)2, Phosphine, K2CO3, DMA, 130℃, rt to N2, 4 hrs; (e) EtOH, NaBH4, rt to N2, 4 hrs.

[0015] Specifically, the preparation method of the compound 2-(2-bromophenyl)-N-(3,4-dimethoxyphenethyl)acetamide shown in Formula 1 is as follows: 2-bromophenylacetic acid is dissolved in dichloromethane (DCM), 1-hydroxybenzotriazole (HOBT), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 3,4-dimethoxyphenethylamine and triethylamine (ET3N) are added to react to obtain the compound shown in Formula 1; the chemical reaction formula is as follows:

[0016]

[0017] The preparation method of the compound 1-(2-bromobenzyl)-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline shown in Formula 2 is as follows: dissolving the compound shown in Formula 1 with DCM, adding phosphorus pentachloride (PCl5), refluxing the reaction, then drying the DCM, adding MeOH and sodium borohydride, and reacting to obtain the compound shown in Formula 2; specifically, the reflux reaction time is 10-30 minutes; further, the reflux reaction time is 20 minutes; the chemical reaction formula is as follows:

[0018]

[0019] The preparation method of the compound 1-(1-(2-bromobenzyl)-6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethane-1-one shown in Formula 3 is as follows: dissolving the compound shown in Formula 2 with DCM, adding trifluoroacetic anhydride (TFAH) and Et3N, and reacting to obtain the compound shown in Formula 3; specifically, the reaction temperature is 0-5°C, and the reaction time is 1-3h; further, the reaction temperature is 0°C, and the reaction time is 2h; the chemical reaction formula is as follows:

[0020]

[0021] The preparation method of the compound 1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)-2,2,2-trifluoroethane-1-one shown in formula 4 is as follows: after dissolving the compound shown in formula 3 with N,N-dimethylacetamide (DMA), tri(4-fluorophenyl)phosphine ((FC6H4)3P), K2CO3 and palladium acetate (Pd(OAc)2) are added to react to obtain the compound shown in formula 4; specifically, the reaction temperature is 100-150°C and the reaction time is 5-10h; further, the reaction temperature is 130°C and the reaction time is 6h. The chemical reaction formula is as follows:

[0022]

[0023] The preparation method of the compound 1,2-dimethoxy-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline shown in Formula 5 is as follows: dissolve the compound shown in Formula 4 with DCM, add NaBH4, and react to obtain the compound shown in Formula 5. Specifically, the temperature for adding NaBH4 is 0°C, and the reaction time is 3-5h; further, the reaction time is 4h. The chemical reaction formula is as follows:

[0024]

[0025] The compound shown in Formula 5 generates a series of nuciferine derivatives under the action of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), N,N-diisopropylethylamine (DIEPA) or triethylamine: the chemical reaction formula is as follows:

[0026]

[0027] Specifically, when the R group structure is 6a, the preparation method of the compound 2-(2-bromophenyl)-1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)ethan-1-one shown in formula 6a is: dissolving the compound shown in formula 5 with DMF, adding 2-bromophenylacetic acid, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N-diisopropylethylamine (DIEPA), and reacting to obtain the compound shown in formula 6a; specifically, the reaction time is 15-20 hours, and further, the reaction time is 18 hours.

[0028] When the R group structure is 6b, the preparation method of the compound (4-chloromethylphenyl) (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo [de, g] quinolin-6-yl) ketone represented by formula 6b is: dissolving the compound represented by formula 5 with DMF, adding 4-(chloromethyl) benzoic acid, HATU and DIEPA, and reacting to obtain the compound represented by formula 6b; specifically, the reaction temperature is room temperature, the reaction time is 15-20 hours, and further, the reaction time is 18 hours.

[0029] When the R group structure is 6c, the preparation method of the compound represented by formula 6c (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(4-fluorophenyl)methanone is: dissolving the compound represented by formula 5 with DMF, adding 4-fluorobenzoic acid, HATU and DIEPA, and reacting to obtain the compound represented by formula 6c; specifically, the reaction temperature is room temperature, the reaction time is 15-20h, and further, the reaction time is 18h.

[0030] When the R group structure is 6d, the synthesis of the compound represented by formula 6d (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(4-methoxyphenyl)methanone: the compound represented by formula 5 is dissolved in DMF, 4-methoxybenzoic acid, HATU and DIEPA are added, and the compound represented by formula 6d is obtained after reaction; specifically, the reaction temperature is room temperature, the reaction time is 15-20h, and further, the reaction time is 18h.

[0031] When the R group structure is 6g, the synthesis of the compound 1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)prop-2-en-1-one shown in formula 6g: dissolve the compound shown in formula 5 in DMF, add acrylic acid, HATU and DIEPA, and react to obtain the compound shown in formula 6g; specifically, the reaction temperature is room temperature, the reaction time is 15-20h, and further, the reaction time is 18h.

[0032] When the R group structure is 6h, the synthesis of the compound represented by formula 6h (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(2-methylfuran-3-yl)methanone: dissolve the compound represented by formula 5 in DMF, add 2-methylfuran-3-carboxylic acid, HATU and DIEPA, and react to obtain the compound represented by formula 6h; specifically, the reaction temperature is room temperature, the reaction time is 15-20h, and further, the reaction time is 18h.

[0033] When the R group structure is 6e, the synthesis of the compound represented by formula 6e (1,2-dimethoxy-6-toluenesulfonyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline: the compound represented by formula 5 is dissolved in DCM, and p-toluenesulfonyl chloride and Et3N are added to react to obtain the compound represented by formula 6e; specifically, the reaction temperature is room temperature, the reaction time is 20-25h, and further, the reaction time is 24h.

[0034] When the R group structure is 6f, the synthesis of the compound represented by formula 6f (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(pyrrolidin-1-yl)methanone: dissolve the compound represented by formula 5 in DMF, add pyrrolidine-1-carbonyl chloride, potassium carbonate and potassium iodide, and react to obtain the compound represented by formula 6f; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0035] When the R group structure is 6i, the synthesis of the compound 1,2-dimethoxy-6-(methylsulfonyl)-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline represented by formula 6i: the compound represented by formula 5 is dissolved in DCM, and methylsulfonyl chloride and Et3N are added to react to obtain the compound represented by formula 6i; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0036] When the R group structure is 6j, the synthesis of the compound represented by formula 6j ((1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(4-methylpiperazine-1-yl)methanone): the compound represented by formula 5 is dissolved in DCM, and 4-methylpiperazine-1-carbonyl chloride, K2CO3 and KI are added and reacted to obtain the compound represented by formula 6j; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0037] When the R group structure is 6l, the synthesis of the compound shown in formula 6l ((1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-yl)(morpholinyl)methanone): the compound shown in formula 5 is dissolved in DCM, and morpholine-4-carbonyl chloride, K2CO3 and KI are added and reacted to obtain the compound shown in formula 6l; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0038] When the R group structure is 6m, the synthesis of the compound represented by formula 6m (N,N-diethyl-1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-carboxamide): the compound represented by formula 5 is dissolved in DCM, and diethylcarbamoyl chloride, K2CO3 and KI are added to react to obtain the compound represented by formula 6m; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0039] When the R group structure is 6n, the synthesis of the compound represented by formula 6n (N,N-dimethyl-1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-carboxamide): the compound represented by formula 5 is dissolved in DCM, and dimethylcarbamoyl chloride, K2CO3 and KI are added and reacted to obtain the compound represented by formula 6n; specifically, the reaction temperature is room temperature, the reaction time is 40-50h, and further, the reaction time is 48h.

[0040] When the R group structure is 6k, the synthesis of the compound shown in formula 6k (1,2-dimethoxy-6-(4-methoxybenzyl)-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline): the compound shown in formula 5 is dissolved in DCM, and p-methoxybenzaldehyde and sodium triacetylborohydride are added and reacted to obtain the compound shown in formula 6k. Specifically, the reaction temperature is room temperature, the reaction time is 20-30 hours, and further, the reaction time is 24 hours.

[0041] In another aspect, the present invention provides the use of the nuciferine derivative prepared by the above-mentioned preparation method in the preparation of drugs for treating arrhythmia.

[0042] Specifically, the arrhythmia includes sinus arrhythmia, atrial arrhythmia, supraventricular arrhythmia and ventricular arrhythmia.

[0043] Furthermore, the sinus arrhythmia includes sinus bradycardia, sinus tachycardia, sinus arrhythmia, sinus arrest and sinoatrial block; the atrial arrhythmia includes atrial premature beats, atrial tachycardia, and atrial premature contractions; the supraventricular arrhythmia includes paroxysmal supraventricular tachycardia; the ventricular arrhythmia includes ventricular premature beats, ventricular tachycardia, and ventricular fibrillation.

[0044] Furthermore, the arrhythmia is arrhythmia induced by barium chloride or arrhythmia induced by aconitine.

[0045] Preferably, the R group structure of the nuciferine derivative is 6a.

[0046] Specifically, the nuciferine derivative is used as an active ingredient in medicine.

[0047] Specifically, the dosage form of the drug is drops, mixtures, tinctures, injections, tablets, powders, oral liquids, capsules, granules, ointments, suspensions, powders, emulsions, solutions, pellets, pills, buccal tablets, lyophilized powder injections, gels, suppositories or aerosols.

[0048] Furthermore, the dosage form of the drug is an injection.

[0049] Specifically, the dosage of the drug is 1.5-15 mg / kg based on the dosage of the nuciferine derivative;

[0050] Furthermore, the dosage of the drug is 2.5-10 mg / kg based on the dosage of the nuciferine derivative.

[0051] In another aspect, the present invention provides the use of the nuciferine derivative prepared by the above-mentioned preparation method in the preparation of a drug for treating QT syndrome.

[0052] Specifically, the QT syndrome includes long QT syndrome and short QT syndrome.

[0053] Preferably, the R group structure of the nuciferine derivative is 6a.

[0054] Furthermore, the dosage form of the drug is an injection.

[0055] The beneficial effects of the present invention are:

[0056] Compared with existing products, the nuciferine derivatives provided by the present invention greatly improve the antiarrhythmic therapeutic effect, reduce adverse drug reactions, and reduce the risk of inducing ventricular arrhythmias. In addition, the raw materials are cheap and easily available, which greatly reduces the production cost and is suitable for large-scale production.

[0057] The nuciferine derivatives of the present invention can shorten the time for recovery from arrhythmia, prolong the time after the heart rate returns to normal, reduce the heart rate, shorten the QT interval time and QRS time, increase the RR interval time, QTc, R voltage, increase the time of ventricular premature beats, ventricular tachycardia, ventricular fibrillation and cardiac arrest, can significantly reduce the amplitude of action potential and resting potential, have strong affinity in the process from inactivation to activation of Nav channel and Cav channel, and have a certain therapeutic effect on short QT syndrome and long QT syndrome. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 is the hydrogen spectrum of the compound of formula 1.

[0059] Figure 2 It is the carbon spectrum of the compound of formula 1.

[0060] Figure 3 It is the hydrogen spectrum of the compound of formula 2.

[0061] Figure 4 It is the carbon spectrum of the compound of formula 2.

[0062] Figure 5 is the hydrogen spectrum of the compound of formula 3.

[0063] Figure 6 It is the carbon spectrum of the compound of formula 3.

[0064] Figure 7 It is the hydrogen spectrum of the compound of formula 4.

[0065] Figure 8 It is the carbon spectrum of the compound of formula 4.

[0066] Fig. 9 is the hydrogen spectrum of the compound of formula 5.

[0067] Fig.10 It is the carbon spectrum of the compound of formula 5.

[0068] Fig.11 is the hydrogen spectrum of the compound of formula 6a.

[0069] Fig.12 This is the carbon spectrum of the compound of formula 6a.

[0070] Fig.13 is the hydrogen spectrum of the compound of formula 6b.

[0071] Fig.14 It is the carbon spectrum of the compound of formula 6b.

[0072] Fig.15 is the hydrogen spectrum of the compound of formula 6c.

[0073] Fig.16 This is the carbon spectrum of the compound of formula 6c.

[0074] Fig.17 is the hydrogen spectrum of the compound of formula 6d.

[0075] Fig.18 This is the carbon spectrum of the compound of formula 6d.

[0076] Fig.19 is the hydrogen spectrum of the compound of formula 6e.

[0077] Fig. 20 This is the carbon spectrum of the compound of formula 6e.

[0078] Fig.21 is the hydrogen spectrum of the compound of formula 6f.

[0079] Fig. 22 This is the carbon spectrum of the compound of formula 6f.

[0080] Fig.23 This is the hydrogen spectrum of the compound of formula 6g.

[0081] Fig.24 This is the carbon spectrum of the compound of formula 6g.

[0082] Fig.25 is the hydrogen spectrum of the compound of formula 6h.

[0083] Fig.26 This is the carbon spectrum of the compound of formula 6h.

[0084] Fig. 27 is the hydrogen spectrum of the compound of formula 6i.

[0085] Fig.28 is the carbon spectrum of the compound of formula 6i.

[0086] Fig.29 is the hydrogen spectrum of the compound of formula 6j.

[0087] Fig.30 This is the carbon spectrum of the compound of formula 6j.

[0088] Fig.31 This is the hydrogen spectrum of the compound of formula 6k.

[0089] Fig.32 This is the carbon spectrum of the compound of formula 6k.

[0090] Fig.33 is the hydrogen spectrum of the compound of formula 61.

[0091] Fig.34 This is the carbon spectrum of the compound of formula 61.

[0092] Fig.35 is the hydrogen spectrum of the compound of formula 6m.

[0093] Fig.36 This is the carbon spectrum of the compound of formula 6m.

[0094] Fig.37 is the hydrogen spectrum of the compound of formula 6n.

[0095] Fig.38 This is the carbon spectrum of the compound of formula 6n.

[0096] Fig.39 The effects of nuciferine derivatives on aconitine-induced arrhythmia (n=6); in the figure, A is the amount of aconitine required for inducing VP in rats; B is the amount of aconitine required for inducing VT in rats; C is the amount of aconitine required for inducing VF in rats; D is the amount of aconitine required for inducing CA in rats; compared with NS: #P<0.05, ##P<0.01, ###P<0.001; compared with DMSO: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001; compared with Verapamil: +P<0.05, ++P<0.01, +++P<0.001; compared with Nucifenrine: *P<0.05, **P<0.01, ***P<0.001.

[0097] Fig.40 The effect of different concentrations of 6a on the recovery time of BaCl2-induced arrhythmia (n=6), compared with NS: ###P<0.001.

[0098] Fig.41 The effects of different concentrations of 6a on the normal rhythm maintenance time induced by BaCl2 (n=6); compared with NS: ###P<0.001; compared with DMSO: ▲P<0.05, ▲▲▲P<0.001.

[0099] Fig.42 The effects of 6a on the electrocardiographic parameters of normal rats (n=6); in the figure, A is the effect of 6a on HR of rats, B is the effect of 6a on RR of rats, C is the effect of 6a on QT of rats, D is the effect of 6a on QTc of rats, E is the effect of 6a on R voltage of rats, and F is the effect of 6a on QRS of rats; compared with 30s: *P<0.05, **P<0.01, ***P<0.001.

[0100] Fig.43The figure shows the effect of the compound represented by formula 6a on APD of rat cardiomyocytes (n=9). In the figure, A is the effect of 6a at a concentration of 10 μM on APD of rat cardiomyocytes; B is the effect of 6a at a concentration of 10 μM on APA of rat cardiomyocytes; C is the effect of 6a at a concentration of 10 μM on resting potential of rat cardiomyocytes; D is the effect of 6a at a concentration of 10 μM on maximum AP depolarization velocity (Vmax); EG are the effects of 6a at a concentration of 10 μM on repolarization of rat cardiomyocytes to 10%, 50% and 90% (APD10, 50, 90), respectively; compared with the control group: *p<0.05, ***p<0.001.

[0101] Fig.44 The compound represented by formula 6a has an effect on rat myocardial cell I Na Effects of activation, inactivation and recovery; A is the effect of 6a at 10 μM on I Na The effect of steady-state activation current curve; the left side of B is I Na IV relationship diagram; the middle of B is the steady-state activation curve calculated based on the IV curve; the right of B is I Na Typical schematic diagram of Na Steady-state inactivation current trajectory; D is I Na Steady-state inactivation curve; E is I Na Recovery curve current trajectory after inactivation; F is N av Recovery curve after inactivation.

[0102] Fig.45 The compound represented by formula 6a has an effect on rat ventricular myocytes I Ca Effects of activation, inactivation and recovery; A is I Ca Steady-state activation current curve; left in B is I Ca IV relationship diagram; the middle of B is the steady-state activation curve calculated based on the IV curve; the right of B is I Ca Typical schematic diagram of Ca Steady-state inactivation current curve; D is I Ca Steady-state inactivation curve; E is I Ca The current trajectory of the recovery curve after inactivation; F is the recovery curve of Cav after inactivation.

[0103] Fig.46 is the effect of the compound shown in formula 6a on the Nav1.5 sodium current (INa) of HEK293 cells; A is the effect of compound 6a (1 μM-100 μM) on I Na B is the effect of compound 6a on I in the resting state or semi-inactivated state Na Curve C is the inhibitory effect of 6a on Nav1.5 in the resting state or semi-inactivated state (n=4).

[0104] Fig.47 is the effect of the compound shown in Formula 6a on the Cav1.2 channel; A is the H A is the effect of different concentrations of 6a on Cav1.2 channels when A = -60mV; B is the current curve of 6a acting on Cav1.2; C is the inhibitory effect of 6a on Cav1.2 channel current.

[0105] Fig.48 A is the effect of the compound represented by formula 6a on the hERG channel; A is the effect of compound 6a at different concentrations on the hERG channel; B is the IKr current curve after treatment with 6a; C is the inhibitory effect of compound 6a on IKr current; D is the effect of 6a on Ikr current at different concentrations.

[0106] Fig.49 A is the effect of cisapride on hERG channel; A is the effect of different concentrations of cisapride on hERG channel; B is the IKr current curve after cisapride treatment; C is the inhibitory effect of cisapride on IKr current. DETAILED DESCRIPTION

[0107] In order to make the technical means, creative features, objectives and effects of the present invention easy to understand, the present invention is further explained in conjunction with specific embodiments below, but the following embodiments are only preferred embodiments of the present invention, not all. In the following embodiments, unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same.

[0108] Example 1

[0109] Dissolve 2-bromophenylacetic acid (5.00 g) in dichloromethane (DCM), stir for 10 min under ice bath, add 1-hydroxybenzotriazole (HOBT) 3.15 g and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) 4.45 g, react for 20 min under ice bath, add 0.8 equivalent of 3,4-dimethoxyphenylethylamine (3.80 g) and 1.1 equivalent of triethylamine (ET3N) 2.25 g. After reacting for 1 h under ice bath, remove the ice bath, continue to react for 24 h at room temperature, extract with dilute hydrochloric acid aqueous solution with pH 4-5, dry the organic layer with anhydrous sodium sulfate, and concentrate the organic layer. Add DCM to the concentrated product until it is completely dissolved, then add frozen ethyl acetate (EA) until crystals are just precipitated, seal and put in -20°C refrigerator to store crystals. After 24 hours, the solution was filtered to obtain precipitated crystals, which were detected to be the compound of formula 1 with relative purity. The hydrogen spectrum of the compound of formula 1 is as follows: Figure 1 As shown, the carbon spectrum of the compound of formula 1 is as follows Figure 2 shown.

[0110] 1 H NMR(400MHz,CHLOROFORM-d)d ppm 2.73(t,J=6.93Hz,2H)3.46-3.54(m,2H)3.69(s,2H)3.88(s,3H)3.86(s,3H)5.47(br.s.,1H)6.58-6.6 7(m,2H)6.75(d,J=8.17Hz,1H)7.13-7.20(m,1H)7.26-7.29(m,1H)7.30(s,1H)7.57(d,J=8.04Hz,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 35.04,40.81,44.10,55.86,55.96,111.36,111.81,120.61,124.98,127. 97,129.09,131.11,131.68,133.10,134.82,147.66,149.04,169.48; HRMS m / z calcd for[C 18 H 20 BrNO3+H] + 380.0627, found 380.0657.

[0111] Example 2

[0112] Dissolve the compound of formula 1 (3.50 g) in DCM, add 3 times the amount of phosphorus pentachloride (PCl5) 5.80 g in an ice bath, react in an ice bath for 10 minutes, then remove the ice bath, heat to reflux for 2 hours, adjust the pH to neutral with saturated sodium bicarbonate (NaHCO3), extract with water and DCM, dry the organic layer with anhydrous sodium sulfate, and concentrate on a rotary evaporator to obtain an oily substance. Add methanol to dissolve the spin-dried oily substance, add 3 times the amount of sodium borohydride (NaBH4) 1.05 g in small amounts and multiple times at 0°C, react at 0°C for 30 minutes, and then react overnight at room temperature. Spin the reaction solution to dryness, add DCM and water for extraction, take the organic layer and dry it with anhydrous sodium sulfate, and purify it with a chromatographic column to obtain the compound of formula 2 (2.68 g) with a yield of 79.57%. The hydrogen spectrum of the compound of formula 2 is shown as follows Figure 3 As shown, the carbon spectrum of the compound of formula 2 is as follows Figure 4 shown.

[0113] 1H NMR(400MHz,CHLOROFORM-d)d ppm 1.75(br.s.,1H)2.72-2.84(m,2H)2.96-3.09(m,2H)3.29(dt,J=12.43,6.15Hz,1H)3.39(dd,J=13.61,3.84Hz,1H)3.89(s,3H)3.85 (s,3H)4.30(dd,J=9.90,3.71Hz,1H)6.63(s,1H)6.76(s,1H)7.15(dt,J=8.41,4.33Hz,1H)7.29-7.32(m,2H)7.63(d,J=7.92Hz,1H); 13 CNMR(101MHz,CHLOROFORM-d)d ppm29.47,40.06,43.23,54.92,55.88,55.97,109.76,111.77,124.97,127 .22,127.44,128.24,130.60,132.02,133.10,138.89,147.10,147.57; HRMS m / z calcd for[C 18 H 20 BrNO2+H] + 362.0667, found 362.0677.

[0114] Example 3

[0115] Dissolve 2.68g of the compound of formula 2 in DCM, add 1.2 times the amount of trifluoroacetic anhydride (TFAH) 1.86g and 2.5 times the amount of Et3N (1.87g) in an ice bath. After reacting at 0°C for 2h, extract with a dilute hydrochloric acid solution of pH=5-6, take the organic layer, extract again with saturated sodium chloride (NaCl) and saturated NaHCO3 solution, take the organic layer and dry it with anhydrous NaSO4, concentrate it, and purify it with a chromatographic column to obtain the product of formula 3 (2.58g) with a yield of 75.68%. The hydrogen spectrum of the compound of formula 3 is as follows Figure 5 As shown, the carbon spectrum of the compound of formula 3 is as follows Figure 6 shown.

[0116] 1H NMR(400MHz,CHLOROFORM-d)d ppm 2.83(dt,J=16.11,3.51Hz,1H)2.98(ddd,J=16.24,11.04,5.51Hz,1H)3.21( dd,J=13.61,8.78Hz,1H)3.39-3.47(m,1H)3.75(s,3H)3.77-3.83(m,1H)3.8 8(s,3H)4.02-4.12(m,1H)5.81(dd,J=8.66,5.94Hz,1H)6.49(s,1H)6.60-6. 65(m,1H)7.09-7.18(m,2H)7.20-7.27(m,1H)7.56(dd,J=7.92,0.99Hz,1H); 13 CNMR(101MHz,CHLOROFORM-d)d ppm 28.77,40.02,40.06,41.55,53.83,55.85,55.92,110.08,111.06,124.86,12 5.40,126.62,127.36,128.71,131.48,132.86,136.54,147.71,148.32; HRMS m / z calcd for[C 20 H 19 BrF3NO3+H] + 460.0450, found 460.0500.

[0117] Example 4

[0118] Dissolve 1 times the amount of compound 3 (2.58 g) in N,N-dimethylacetamide (DMA), add 0.2 times the amount of tri(4-fluorophenyl)phosphine ((FC6H4)3P) 0.35 g and 1 times the amount of K2CO3 (0.78 g), react at 130°C for 3 hours under N2 protection, then add 0.1 times the amount of palladium acetate (Pd(OAc)2) 0.13 g to continue the reaction. After 3 hours, after TLC detection, the reaction solution was filtered with silica gel, extracted with water and DCM, and purified by chromatographic column to obtain 1.83 g of compound 4, with a yield of 89.29%. The hydrogen spectrum of compound 4 is shown as follows: Figure 7 As shown, the carbon spectrum of the compound of formula 4 is as follows Figure 8 shown.

[0119] 1 H NMR(400MHz,CHLOROFORM-d)ppm 2.76(d,J=15.65Hz,1H)2.83

[0120] -2.91(m,1H)2.94(dd,J=12.53,3.73Hz,1H)3.06(dd,J=13.45,4.03Hz,1H)3.30

[0121] -3.41(m,1H)3.68(s,3H)3.90(s,3H)4.18-4.27(m,1H)5.03(dd,J=13.75,3.85 Hz,1H)6.67(s,1H)7.23-7.26(m,1H)7.30-7.38(m,1H)8.45(d,J=7.82Hz,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 28.79,30.39,33.58,52.29,53.83,55.85,55.94,56.03,60.10,111.28,127.35 ,128.05,128.39,128.56,128.64,128.73,131.31,131.48,132.90,135.77; HRMS m / z calcd for[C 20 H 18 F3NO3+Na] + 400.1239, found 400.1205.

[0122] Example 5

[0123] EtOH was added to 1 times the amount of the compound of formula 4 (1.83 g) to dissolve it, and 20 times the amount of NaBH4 (3.82 g) was added in small amounts at 0°C for several times. The mixture was reacted at 0°C for 4 hours, and extracted with a dilute hydrochloric acid aqueous solution with a pH of 5-6 and DCM after TLC detection. The organic layer was dried with anhydrous Na2SO4, and purified by a chromatographic column. The yield of the compound of formula 5 (1.13 g) was 79.89%. The hydrogen spectrum of the compound of formula 5 is shown in FIG. Fig. 9 As shown, the carbon spectrum of the compound of formula 5 is as follows Fig.10 shown.

[0124] 1 H NMR(400MHz,CHLOROFORM-d)ppm 2.65-2.80(m,3H)2.81-2.88(m,1H)2.97(s,1H)3.01(d,J=8.68Hz,2H)3.34-3.40(m,1H)3.67(s,3H)3. 89(s,3H)6.60(d,J=3.42Hz,1H)6.65(s,1H)7.21-7.25(m,2H)7.29-7.34(m,1H)8.39(d,J=7.82Hz,1H); 13C NMR(101MHz,METHANOL-d4)d ppm 27.23,35.72,42.09,53.19,55.01,59.19,111.74,126.22,126.30,126.8 0,127.39,127.61,128.03,128.17,131.78,135.18,145.25,152.65; HRMS m / z calcd for[C 18 H 19 NO2+H] + 282.1416, found 282.1497.

[0125] Example 6

[0126] Dissolve 0.20g of 1-fold amount of compound 5 in DMF for later use. Add 0.18g of 1.5-fold amount of 2-bromophenylacetic acid and 0.40g of 1.8-fold amount of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) in a 25ml reaction bottle and dissolve in DMF. Stir and react at room temperature for 0.5h. Then, drop the compound of formula V dissolved in DMF into the reaction bottle and continue to add 0.14g of 1.8-fold amount of N,N-diisopropylethylamine (DIEPA). React at room temperature for 18h under N2. After TLC detection, dry on a rotary evaporator and purify by a normal silica gel column to obtain compound 6a (0.23g) with a yield of 73.15%. The hydrogen spectrum of compound 6a is shown in Fig.11 As shown, the carbon spectrum of the compound of formula 6a is as follows Fig.12 shown.

[0127] 1 H NMR(400MHz,DMSO-d6)d ppm 2.70-3.04(m,4H)3.23-3.33(m,1H)3.61(s,3H)3.86(s,3H)3.94-4.06(m,2H)4.19-4.27(m,1H)4.79-4.91(m,1 H)6.91-6.98(m,1H)7.16-7.23(m,1H)7.27-7.32(m,2H)7.36-7.41(m,3H)7.55-7.68(m,1H)8.23-8.37(m,1H); 13C NMR(101MHz,DMSO-d6)d ppm30.49,34.09,41.16,41.36,50.62,56.30,60.05,112.75,125.19,126.10,126.98,127.35,128.07,128. 16,128.38,128.89,129.12,130.01,131.71,132.29,132.70,136.63,137.27,145.51,152.19,168.46; HRMS m / z calcd for[C 26 H 24 BrNO3+Na] + 500.0950, found 500.0917.

[0128] Example 7

[0129] In a 25 ml reaction bottle, add 1.5 times the amount of 4-(chloromethyl)benzoic acid (0.18 g), 1 times the amount of the compound of formula 5 (0.20 g), 1.8 times the amount of HATU (0.40 g), and 1.8 times the amount of DIEPA (0.14 g). The other operations are the same as those in Example 6. After purification by chromatographic column, the compound of formula 6b (0.25 g) is obtained with a yield of 81.52%. The hydrogen spectrum of the compound of formula 6b is shown in Fig.13 As shown, the carbon spectrum of the compound of formula 6b is as follows Fig.14 shown.

[0130] 1 H NMR(400MHz,CHLOROFORM-d)ppm 2.64(d,J=15.41Hz,1H)2.86-3.01(m,2H)3.12(br.s.,1H)3.24(d,J=12.72Hz,1H)3.68(s,3H)3.90(s,3H)4.59-4.64(m,3H)6. 67(s,1H)7.22-7.25(m,1H)7.29-7.35(m,1H)7.46(s,3H)7.49(d,J=8.31Hz,1H)8.09(d,J=8.31Hz,1H)8.44(d,J=7.82Hz,1H); 13CNMR(101MHz,CHLOROFORM-d)d ppm 19.17,30.72,45.30,45.62,56.01,60.04,77.23,111.39,126.09,127.11,127.17,127.55,127.84,128.4 6,128.56,128.78,129.28,130.61,130.92,131.52,136.47,136.82,138.87,142.98,145.93,152.25; HRMS m / z calcd for[C 26 H 24 ClNO3+H] + 434.1455, found 434.1513.

[0131] Example 8

[0132] In a 25 ml reaction bottle, add 1.5 times the amount of 4-fluorobenzoic acid (0.15 g), 1 times the amount of the compound of formula 5 (0.20 g), 1.8 times the amount of HATU (0.40 g), and 1.8 times the amount of DIEPA (0.14 g). The other operations are the same as those in Example 6. After purification by a normal silica gel column, the compound of formula 6c (0.19 g) is obtained with a yield of 68.35%. The hydrogen spectrum of the compound of formula 6c is shown in FIG. Fig.15 As shown, the carbon spectrum of the compound of formula 6c is as follows Fig.16 shown.

[0133] 1 H NMR(600MHz,CHLOROFORM-d)d ppm 2.65(d,J=15.44Hz,1H)2.86-2.97(m,2H)3.25(br.s.,1H)3.68(s,3H)3.90(s,3H)6.67(s,1H)7.11(t, J=8.45Hz,2H)7.22-7.25(m,1H)7.30-7.36(m,1H)7.46(dd,J=8.36,5.45Hz,2H)8.44(d,J=7.81Hz,1H); 13 C NMR(151MHz,CHLOROFORM-d)d ppm 18.42,30.68,55.99,60.03,111.36,115.58,115.72,126.04,127.11,127.84,128.46,128. 93,128.98,131.50,132.78,132.80,136.44,145.91,152.24,162.47,164.13,169.91; HRMS m / z calcd for[C25 H 22 FNO3+H] + 405.1617, found 405.1681.

[0134] Example 9

[0135] In a 50 ml reaction bottle, add 1.5 times the amount of 4-methoxybenzoic acid (0.16 g), 1 times the amount of the compound of formula 5 (0.20 g), 1.8 times the amount of HATU (0.40 g), and 1.8 times the amount of DIEPA (0.14 g). The other operations are the same as those in Example 6. The compound of formula 6d (0.21 g) is purified by chromatographic column with a yield of 73.36%. The hydrogen spectrum of the compound of formula 6d is as follows: Fig.17 As shown, the carbon spectrum of the compound of formula 6d is as follows Fig.18 shown.

[0136] 1 H NMR(400MHz,CHLOROFORM-d)d ppm 2.64(d,J=15.41Hz,1H)2.87-2.98(m,2H)3.14(d,J=11.37Hz,1H)3.20-3.31(m,1H)3.68(s,3H)3.81-3.85(m,3H) 3.90(s,3H)6.67(s,1H)6.90-6.96(m,2H)7.24(s,1H)7.26-7.35(m,2H)7.40-7.47(m,2H)8.44(d,J=7.70Hz,1H); 13 C NMR(151MHz,CHLOROFORM-d)d ppm 18.44,29.70,30.77,34.75,55.36,55.99,58.48,60.03,111.34,113.80,126.40,127.03,127.79,12 7.85,128.42,128.51,128.68,128.98,129.17,131.56,136.66,145.84,152.15,160.62,170.74; HRMS m / z calcdfor[C 26 H 25 NO4+H] + 417.1817, found 417.1876.

[0137] Example 10

[0138] Add 1 times the amount of compound 5 (0.15 g) to a 25 ml reaction bottle, dissolve it in DCM, add 2.5 times the amount of Et3N (0.13 g) at 0°C, add 1.1 times the amount of p-toluenesulfonyl chloride (0.11 g) after 10 minutes, react at 0°C under N2. After 15 hours, TLC detection, extract with saturated NaHCO3 solution and DCM, take the organic layer and dry it with anhydrous Na2SO4, and concentrate to obtain a yellow oil. After purification by chromatography, the compound 6e (0.13 g) is obtained with a yield of 55.32%. The hydrogen spectrum of the compound 6e is as follows Fig.19 As shown, the carbon spectrum of the compound of formula 6e is as follows Fig. 20 shown.

[0139] 1 H NMR(400MHz,CHLOROFORM-d)d ppm 2.41(s,3H)2.42-2.54(m,2H)3.05(t,J=13.79Hz,1H)3.20(dd,J=13.92,4 .27Hz,1H)3.32(ddd,J=14.13,11.47,3.09Hz,1H)3.67(s,3H)3.88(s,3H) 4.08-4.19(m,1H)4.63(dd,J=13.79,4.14Hz,1H)6.57(s,1H)7.27(d,J=7. 92Hz,2H)7.30-7.40(m,3H)7.73(d,J=8.29Hz,2H)8.44(d,J=7.92Hz,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm21.52(s,1C)28.80(s,1C)37.92(s,1C)40.96,55.91,60.08,111.41,125.38,126.94,127.18 ,127.93,128.43,128.54,128.93,129.90,131.48,136.50,137.98,143.32,145.75,152.26; HRMS m / z calcd for[C 25 H 25 NO4S+Na] + 458.1504,found458.1396.

[0140] Embodiment 11

[0141] In a 25 ml reaction bottle, add 1 times the amount of compound 5 of formula 5 (0.06 g), 1.2 times the amount of pyrrolidine-1-carbonyl chloride (0.05 g), 1.2 times the amount of potassium carbonate (0.06 g), and 4 times the amount of potassium iodide (0.14 g), and completely dissolve them with an appropriate amount of DMF, protect with N2, and react at room temperature for 48 hours. After TLC detection, the reaction solution is extracted 3 times with ethyl acetate and water, the organic layer is dried over anhydrous Na2SO4, and purified by a chromatographic column to obtain a compound of formula 6f (0.05 g) with a yield of 63%. The hydrogen spectrum of the compound of formula 6f is as follows Fig.21 As shown, the carbon spectrum of the compound of formula 6f is as follows Fig. 22 shown.

[0142] 1 H NMR(400MHz,METHANOL-d4)ppm 2.57-2.66(m,1H)2.72(s,2H)2.82-2.91(m,1H)3.19-3.22(m,5H)3.28(d,J=6.11Hz,3 H)3.52(s,3H)3.78(s,3H)4.48(s,1H)6.71(s,1H)7.10-7.21(m,3H)8.19-8.26(m,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 11.99,12.05,12.24,18.99,19.04,19.40,21.09,23.08,28.92,29.70,31.66,31. 92,36.52,37.27,42.23,42.30,50.18,51.25,71.83,121.72,138.31,121.72; HRMS m / z calcd for[C 23 H 26 N2O3+H] + 401.1943, found 401.1905.

[0143] Example 12

[0144] In a 25 ml reaction bottle, 1 times the amount of the compound of formula 5 (0.10 g), 1.5 times the amount of acrylic acid (0.04 g), 1.8 times the amount of HATU (0.21 g), and 1.8 times the amount of DIEPA (0.07 g) were dissolved in DMF. The other operations were the same as those in Example 6. After purification by silica gel chromatography, the compound of formula 6g (0.08 g) was obtained with a yield of 66.43%. The hydrogen spectrum of the compound of formula 6g is shown in FIG. Fig.23 As shown, the carbon spectrum of the compound of formula 6g is as follows Fig.24 shown.

[0145] 1H NMR(400MHz,CHLOROFORM-d)ppm 2.65-2.79(m,1H)2.79-2.95(m,2H)3.67(s,3H)3.85(d,J=13.94Hz,1H)3.90(s,3H)5.69-5.75(m ,1H)6.63(br.s.,1H)6.69(s,1H)7.26-7.30(m,2H)7.33(d,J=7.58Hz,1H)8.45(d,J=7.95Hz,1H); 13 C NMR(151MHz,CHLOROFORM-d)d ppm 27.94,35.52,43.10,55.54,55.92,55.98,60.04,109.78,111.39,124.82,12 6.62,126.85,126.97,127.84,127.91,128.91,132.42,132.75,165.67; HRMS m / z calcd for[C 21 H 21 NO3+H] + 358.1521, found 358.1403.

[0146] Example 13

[0147] Dissolve 1 times the amount of the compound of formula 5 (0.10 g), 1.5 times the amount of 2-methylfuran-3-carboxylic acid (0.07 g), 1.8 times the amount of HATU (0.21 g), and 1.8 times the amount of DIEPA (0.07 g) in DMF. The other operations were the same as those in Example 6. After purification by silica gel chromatography, the compound of formula 6h (0.09 g) was obtained with a yield of 66.55%. The hydrogen spectrum of the compound of formula 6h is shown in FIG. Fig.25 As shown, the carbon spectrum of the compound of formula 6h is as follows Fig.26 shown.

[0148] 1 H NMR(400MHz,CHLOROFORM-d)ppm 2.41(s,3H)2.59(s,1H)2.65-2.74(m,1H)2.90(t,J=12.35Hz,2H)3.25(br.s.,2H)3.65-3.70(m,3H)3.90(s, 3H)6.41(d,J=1.96Hz,1H)6.68(s,1H)7.22-7.25(m,1H)7.27(s,2H)7.30-7.35(m,1H)8.44(d,J=7.82Hz,1H); 13CNMR(101MHz,CHLOROFORM-d)dppm 13.00,30.80,56.01,60.05,77.22,109.78,110.93,111.33,116.34,126.29,127.08,1 27.81,128.45,129.10,131.56,136.54,140.38,145.87,152.19,153.49,165.71; HRMS m / z calcd for[C 24 H 23 NO4+H] + 390.1627, found 390.1685.

[0149] Embodiment 14

[0150] 0.08 g of compound 5 was added to a 25 ml reaction bottle, and 2.5 times the amount of Et3N (0.07 g) was added at 0°C. After 10 minutes, 1.2 times the amount of methylsulfonyl chloride (0.04 g) was added. N2 was reacted at 0°C. After 24 hours, TLC was detected, and the organic layer was extracted with saturated NaHCO3 and DCM. The organic layer was collected, dried with anhydrous Na2SO4, concentrated, and purified by normal silica gel chromatography to obtain yellow-brown needle-shaped crystals of compound 6i (0.04 g). The yield was 40.06%. The hydrogen spectrum of compound 6i is shown in Fig. 27 As shown, the carbon spectrum of the compound of formula 6i is as follows Fig.28 shown.

[0151] 1 H NMR(400MHz,CHLOROFORM-d)2.71(td,J=2.32,15.89Hz,1H),2.83-3.11(m,6H),3.20-3.39(m,1H),3.58-3.71(m,3H),3 .91(s,3H),4.04-4.20(m,1H),4.52(dd,J=5.62,12.23Hz,1H),6.68(s,1H),7.27-7.37(m,3H),8.42(d,J=7.82Hz,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 29.52,37.35,39.67,40.47,52.98,55.98,60.08,111.56,125.24,127.2 6,127.98,128.04,128.49,128.54,128.60,131.36,136.32,152.49; HRMS m / z calcd for[C 19 H 21NO4S+H] + 360.1191, found 360.1253.

[0152] Embodiment 15

[0153] In a 25 ml reaction bottle, add 1 times the amount of the compound of formula 5 (0.10 g) and dissolve it in DCM, 1.2 times the amount of 4-methylpiperazine-1-carbonyl chloride (0.07 g), 4 times the amount of KI (0.23 g), and 1.2 times the amount of K2CO3 (0.06 g). The other operations are the same as those in Example 11. After purification by a chromatographic column, the compound of formula 6j (0.094 g) is obtained with a yield of 68.21%. The hydrogen spectrum of the compound of formula 6j is shown in FIG. Fig.29 As shown, the carbon spectrum of the compound of formula 6j is as follows Fig.30 shown.

[0154] 1 H NMR(400MHz,CHLOROFORM-d)2.30(s,3H),2.38-2.67(m,7H),2.71-2.86(m,2H),2.86-3.12(m,2H),3.13-3.28(m,2H),3.33-3.45(m,1H),3.67(s ,3H),3.91(s,3H),4.30(d,J=9.54Hz,1H),5.03(dd,J=3.55,13.82Hz,1H ),6.69(s,1H),7.19-7.26(m,1H),7.28-7.34(m,1H),8.40-8.51(m,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 29.71,30.96,33.93,41.17,46.07,50.77,53.20,53.39,55.10,55.99,60.03,62.31,77.24,1 11.26,126.39,126.97,127.76,128.38,128.62,129.19,131.50,136.77,152.08,168.33; HRMS m / z calcd for[C 24 H 29 N3O3+K] + 446.2209, found 446.2372.

[0155] Example 16

[0156] In a 25 ml reaction bottle, add 1 times the amount of compound 5 (0.11 g), 1.5 times the amount of p-methoxybenzaldehyde (0.08 g), dissolve it with DCM, add 1.2 times the amount of sodium triacetoxyborohydride (0.1 g), N2, and react at room temperature. After 24 hours, TLC detection, extract with DCM and water, dry the organic layer with anhydrous Na2SO4, concentrate, and purify by chromatographic column to obtain compound 6k (0.12 g), with a yield of 75.63%. The hydrogen spectrum of compound 6k is as follows Fig.31 As shown, the carbon spectrum of the compound of formula 6k is as follows Fig.32 shown.

[0157] 1 H NMR(400MHz,CHLOROFORM-d)ppm 3.63(br.s.,3H)3.81(s,3H)3.88(s,3H)4.09(d,J=6.60Hz,1H)6.62(s,1H)6.91(d,J=7.34Hz,2 H)7.27-7.34(m,2H)7.36(d,J=8.56Hz,2H)7.54(dd,J=5.75,3.30Hz,1H)7.72(d,J=5.62Hz,1H); 13 C NMR(101MHz,CHLOROFORM-d)d ppm 19.20,29.74,35.20,48.74,55.31,55.89,57.95,60.14,60.29,71.83,77.27,111.29,113.77, 113.98,127.10,127.23,127.44,127.85,128.39,130.29,132.18,145.19,152.04,158.84; HRMS m / zcalcd for[C 26 H 27 NO3+H] + 402.1991,found402.2066.

[0158] Embodiment 17

[0159] In a 25 ml reaction bottle, add 1 times the amount of product 5 (0.3 g), 1.2 times the amount of morpholine-4-carbonyl chloride (0.19 g), 1.2 times the amount of K2CO3 (0.18 g), and 4 times the amount of KI (0.70 g), and dissolve them completely with an appropriate amount of DMF. The other operations are the same as those in Example 11, and after purification by chromatography, the compound of formula 61 (0.22 g) is obtained with a yield of 48.11%. The hydrogen spectrum of the compound of formula 61 is shown as follows: Fig.33 As shown, the carbon spectrum of the compound of formula 61 is as follows Fig.34 shown.

[0160] 1 H NMR(400MHz,CHLOROFORM-d)ppm 2.71(t,J=13.51Hz,1H)2.76-2.91(m,2H)3.02(dd,J=13.45,4.03Hz,1H)3.21-3.32(m,3H)3.37-3.45(m,2H)3.67(s,3H)3.68- 3.79(m,5H)3.90(s,3H)4.71(dd,J=13.57,3.91Hz,1H)6.66(s,1H)7.21-7.26(m,2H)7.29-7.35(m,1H)8.40(d,J=7.70Hz,1H); 13 CNMR(101MHz,CHLOROFORM-d)d ppm 18.13,29.19,34.55,43.10,46.05,46.21,51.44,54.94,59.09,65.59,65.71,76.21,110.1 0,125.87,125.99,126.65,127.16,127.31,127.80,130.63,135.57,144.63,151.05,163.0.

[0161] Embodiment 18

[0162] In a 25 ml reaction bottle, add 1 times the amount of compound 5 (0.15 g), 1.2 times the amount of diethylcarbamoyl chloride (0.09 g), 1.2 times the amount of K2CO3 (0.09 g), 4 times the amount of KI (0.35 g), and add DMF to completely dissolve it. The other operations are the same as those in Example 11. After purification by chromatography, the compound of formula 6m (0.10 g) is obtained with a yield of 50.21%. The hydrogen spectrum of the compound of formula 6m is shown in Fig.35 As shown, the carbon spectrum of the compound of formula 6m is as follows Fig.36 shown.

[0163] 1 H NMR(400MHz,CHLOROFORM-d)ppm 1.11-1.20(m,6H)2.65(t,J=13.45Hz,1H)2.76-2.97(m,2H)3.04(dd,J=13.45,3.91Hz,1H)3.13-3.35(m,3H)3.38-3. 53(m,3H)3.66(s,3H)3.90(s,3H)4.48(dd,J=13.45,3.91Hz,1H)6.67(s,1H)7.18-7.35(m,3H)8.38(d,J=7.82Hz,1H); 13C NMR(101MHz,CHLOROFORM-d)d ppm 13.26,13.44,29.94,35.37,41.73,42.29,45.49,53.39,55.95,60.14,77.27,111.14,126.90, 127.56,127.62,127.67,128.16,128.29,128.95,131.75,136.82,145.45,151.97,165.00; HRMS m / z calcd for[C 23 H 28 N2O3+Na] + 403.2100, found 403.2085.

[0164] Embodiment 19

[0165] In a 25 ml reaction bottle, add 1 times the amount of compound 5 (0.26 g), 1.2 times the amount of dimethylaminoformyl chloride (0.12 g), 1.2 times the amount of K2CO3 (0.26 g), 4 times the amount of KI (0.62 g), and add DMF to completely dissolve. The other operations are the same as those in Example 11, and the compound of formula 6n (0.10 g) is obtained after purification by normal silica gel chromatography, with a yield of 30.15%. The hydrogen spectrum of the compound of formula 6n is shown in Fig.37 As shown, the carbon spectrum of the compound of formula 6n is as follows Fig.38 shown.

[0166] 1 H NMR(400MHz,METHANOL-d4)ppm 2.55(t,J=13.57Hz,1H)2.73(s,1H)2.75-2.80(m,1H)2.83(s,6H)2.86(d,J=3.91Hz,1H)3.23-3.28(m,1H)3.38-3.4 7(m,1H)3.52(s,3H)3.78(s,3H)4.43(dd,J=13.45,3.91Hz,1H)6.71(s,1H)7.08-7.25(m,3H)8.22(d,J=7.58Hz,1H); 13 C NMR(101MHz,METHANOL-d4)d ppm 29.53,35.47,37.02,44.44,52.68,55.03,59.05,111.36,126.52,126.55,127 .03,127.32,127.68,127.91,129.23,131.61,136.64,145.37,152.12,165.57.

[0167] Example 20 Therapeutic effects of the compounds represented by formula 4-5 and formula 6a-6n on BaCl2-induced arrhythmia in SD rats

[0168] 10g sodium pentobarbital solution was dissolved in 90ml normal saline to obtain 10% sodium pentobarbital solution for use, 80.00mg BaCl2 was dissolved in 10ml normal saline to obtain 8mg / ml BaCl2 solution for use. 14.81mg verapamil solution was dissolved in 2ml DMSO to obtain 7.41mg / ml verapamil solution for use. 25mg nuciferine and nuciferine derivatives were dissolved in 1.35ml DMSO to obtain 18.52mg / ml nuciferine and nuciferine derivatives solution for use.

[0169] Normal male SD rats weighing 200-220g were randomly divided into groups, each group consisted of 6 rats, including normal saline group (NS group) and DMSO group as blank control group, nuciferine group (5mg / kg) and verapamil group (2mg / kg) as positive control group, and nuciferine derivative group (5mg / kg) as experimental group. After the rats were anesthetized by intraperitoneal injection of anesthetic, the BL-420N biological signal acquisition system was connected to record the normal electrocardiogram for 1min, and then the BaCl2 solution (4mg / kg) was injected sublingually. After 3 minutes, normal saline (0.27ml / kg) and DMSO (0.27ml / kg) were injected sublingually again according to the grouping as blank control group, nuciferine (5mg / kg) and verapamil (2mg / kg) as positive drug control group, and synthetic nuciferine derivative (5mg / kg). The II-lead electrocardiogram (ECG) was observed for 35 minutes under the monitoring of the BL-420N biological signal acquisition and analysis system. The antiarrhythmic activity of nuciferine derivatives was evaluated by recording the time required for the heart rate to return to normal after arrhythmia, the time to maintain normality, heart rate, RR interval, QT interval, QTc interval and R voltage in the ECG.

[0170] Example 20-1 Effect of Nuciferine Derivatives on Recovery Time and Maintenance Time of Arrhythmia Induced by BaCl2

[0171] The experimental results are shown in Table 1. After BaCl2 was injected into rats in the NS group and the DMSO group, the rats would have arrhythmia and it would be difficult to return to normal for a long time. This indicates that BaCl2 can induce arrhythmia in rats and the rats will be in an arrhythmic state for a long time. Compared with the NS group and the DMSO group, all the nuciferine derivatives, nuciferine and verapamil can significantly shorten the time for the rats' heart rate to return to normal (P < 0.001). Among them, 6a, 6c, 6f, 6g, 6j, 6m have a significant trend of shortening the recovery time compared with verapamil and nuciferine. Compared with the NS group, except for 6k, 6l, 6m, 6n, the other derivatives, verapamil and nuciferine can significantly prolong the maintenance time after the rats' heart rate returns to normal (P < 0.05). Compared with the DMSO group, all the compounds, nuciferine and verapamil can significantly prolong the maintenance time after the rats' heart rate returns to normal (P < 0.05). Compared with verapamil and nuciferine, 6a had a significant tendency to prolong the heart rate maintenance time of rats, and among the number of rats with a maintenance time ≥ 20 minutes, only 6 rats in all groups were in 6a.

[0172] Table 1 Effects of nuciferine derivatives on BaCl2-induced arrhythmia in rats (n=6)

[0173]

[0174]

[0175] Compared with the NS group: # P<0.05, ## P<0.01, ### P < 0.001;

[0176] Compared with DMSO group: ▲ P<0.05, ▲▲ P<0.01, ▲▲▲ P<0.001.

[0177] Example 20-2 Effect of Nuciferine Derivatives on BaCl2 Changes in Heart Rate (HR) in Rats

[0178] As shown in Table 2, the heart rate of rats significantly increased after BaCl2 injection compared with the normal group, because barium ions will inhibit the sodium and potassium ion channels on the myocardial cell membrane after entering the body, which will lead to a decrease in potassium ions and an increase in sodium ions in the cells, which will induce increased cardiac autonomicity and increased myocardial sympathetic nerve excitement, leading to an increase in heart rate. Injection of verapamil, nuciferine and nuciferine derivatives can reduce heart rate, indicating that nuciferine derivatives have a certain therapeutic effect on the increase in heart rate caused by BaCl2.

[0179] Table 2 Effects and significance of nuciferine derivatives on HR of rats (n=6)

[0180]

[0181]

[0182] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0183] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0184] Example 20-3 Effect of Nuciferine Derivatives on Changes in QT Interval in Rats by BaCl2

[0185] As shown in Table 3, compared with the normal group, the QT interval of rats after BaCl2 injection was significantly reduced, thus inducing Short QT Syndrome (SQTS), a syndrome characterized by shortened QT interval and causing sudden arrhythmic death. Compared with the normal group, the QT interval of all compounds except verapamil and the model group of nuciferine was significantly shortened (P < 0.05). After the injection of nuciferine derivatives and NS, DMSO, verapamil, and nuciferine, except for the QT interval of NS and DMSO, the QT intervals of the other groups were significantly increased (P < 0.05) and tended to return to the normal QT interval, which indicates that the synthesized nuciferine derivatives and nuciferine have a certain therapeutic effect on SQTS caused by BaCl2.

[0186] Table 3 Effects and significance of nuciferine derivatives on QT interval in rats (n=6)

[0187]

[0188]

[0189] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0190] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0191] Example 20-4 Effect of Nuciferine Derivatives on the Change of RR Interval in Rats

[0192] As shown in Table 4, after the rats were injected with BaCl2, the RR interval of normal rats showed a significant downward trend. After the injection of nuciferine derivatives, verapamil and nuciferine, the RR interval of rats showed a significant upward trend. Among them, the RR intervals of 6a, 6b, 6d, 6g, 6h and 6n were significantly increased compared with those of the model group (P < 0.05), and the increased RR intervals were not significantly changed compared with those of the normal group.

[0193] Table 4 Effects and significance of nuciferine derivatives on RR interval in rats (n=6)

[0194]

[0195]

[0196] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0197] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0198] Example 20-5 Effect of Nuciferine Derivatives on the Change of QTc Interval in Rats by BaCl2

[0199] BaCl2 injection will cause the RR interval to shorten and the heart rate to increase, and the increased heart rate will also cause the QT interval to shorten. The present invention investigates whether BaCl2 injection will cause SQTS and whether nuciferine derivatives have a therapeutic effect on SQTS caused by BaCl2 by statistically analyzing the QT interval corrected for heart rate, that is, QTc. The results are shown in Table 5. Normal rats will cause SQTS after BaCl2 injection (P < 0.05). After administration, except for nuciferine, 6e, 6g, 6j, and 6n, the QTc values ​​of the other nuciferine derivatives and verapamil are significantly increased (P < 0.05), especially 4, 5, 6a, 6b, 6c, 6d, 6f, 6g, 6i, 6k, and 6m. The increased QTc values ​​are not significantly different from those of the normal group, which indicates that the above compounds have a certain corrective effect on SQTS caused by BaCl2 injection.

[0200] Table 5 Effects and significance of nuciferine derivatives on QTc interval in rats (n=6)

[0201]

[0202]

[0203] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0204] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0205] Example 20-6 Effect of Nuciferine Derivatives on the Change of BaCl2 to R Voltage in Rats

[0206] The effects of nuciferine derivatives on R voltage of rats are shown in Table 6. After injection of BaCl2, the R voltage of rats decreased significantly (P < 0.05). After injection of verapamil and nuciferine, the R voltage increased significantly compared with the model group (P < 0.05) and had no significant difference from the normal group (P > 0.05). After injection of nuciferine derivatives, the R voltage of the drug-treated groups except 5, 6f, 6i, 6j, 6l, and 6n increased significantly compared with the model group (P < 0.05), and had no significant difference compared with the normal group (P > 0.05).

[0207] Table 6 Effects and significance of nuciferine derivatives on R voltage in rats (n=6)

[0208]

[0209]

[0210] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0211] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0212] Example 20-7 Effect of Nuciferine Derivatives on BaCl2-induced QRS Changes in Rats

[0213] The effects of nuciferine derivatives on QRS in rats are shown in Table 7. After the injection of barium chloride, the QRS time of rats except NS, 5, and 6f was significantly prolonged (P < 0.05). After the injection of nuciferine, nuciferine derivatives and verapamil, the QRS time of the other groups except 6b and 6h had no significant difference compared with the drug-treated group (P > 0.05). This indicates that nuciferine and nuciferine derivatives have a certain therapeutic effect on the prolongation of QRS time caused by the injection of barium chloride.

[0214] Table 7 Effects and significance of nuciferine derivatives on QRS in rats (n=6)

[0215]

[0216]

[0217] Compared with the normal group: #P<0.05, ##P<0.01, ###P<0.001;

[0218] Compared with the model group: ▲P<0.05, ▲▲P<0.01, ▲▲▲P<0.001.

[0219] Example 21 Therapeutic effects of some nuciferine derivatives on aconitine-induced arrhythmia in rats

[0220] After exploring the effects of nuciferine derivatives on aconitine-induced arrhythmia in rats, the present invention screened 4, 5, and 6a to explore the effects of the above compounds on aconitine-induced arrhythmia in rats. The injection concentrations of 4 and 5 were 5 mg / kg, and the concentrations of 6a were 1 mg / kg, 2.5 mg / kg, 5 mg / kg, and 10 mg / kg, respectively.

[0221] Dissolve 5.00 mg of aconitine in 5 mL of normal saline to obtain a 1 mg / ml aconitine stock solution. Take 1 ml of the aconitine stock solution and dilute it to 100 ml with normal saline to obtain a 0.01 mg / ml aconitine solution.

[0222] Healthy male SD rats weighing 200-220g were randomly divided into groups, with 6 rats in each group. The normal saline group and the DMSO group were blank control groups, verapamil and nuciferine were positive drug control groups, and 4, 5, and 6a were experimental groups.

[0223] After intraperitoneal injection of anesthetic, rats were fixed in supine position and connected to BL-420N biological signal acquisition system to record 1 min of normal lead II electrocardiogram (ECG). The blank control group was injected with normal saline (0.27 ml / kg) and DMSO (0.27 ml / kg) sublingually, the positive drug control group was injected with verapamil solution (2 mg / kg) and nuciferine solution (5 mg / kg) sublingually, and the experimental groups were injected with 4 (5 mg / kg), 5 (5 mg / kg), 6a (1 mg / kg), 6a (2.5 mg / kg), 6a (5 mg / kg) and 6a (10 mg / kg) sublingually. Two minutes later, a 0.01 mg / ml aconitine solution was injected into the femoral vein on one side of the rat's thigh at a constant rate of 100 uL / min using a microinjection pump. The ECG was observed and the time of ventricular premature beats (VP), ventricular tachycardia (VT), ventricular fibrillation (VF) and cardiac arrest (CA) in each rat was monitored, and the dosage of aconitine was calculated accordingly.

[0224] The experimental results are as follows Fig.39As shown, compared with verapamil, 5 (5 mg / kg), 6a (5 mg / kg), and 6a (10 mg / kg) can significantly increase the amount of VP induced by aconitine in rats (P < 0.05); compared with verapamil, 5 (5 mg / kg), 6a (5 mg / kg), and 6a (10 mg / kg) can significantly increase the amount of VT induced by aconitine in rats (P < 0.05); compared with verapamil, 4 (5 mg / kg), 5 (5 mg / kg), 6a (2.5 mg / kg), 6a (5 mg / kg), and 6a (10 mg / kg) can significantly increase the amount of VF induced by aconitine in rats (P < 0.05); compared with verapamil, 4, 5, 6a (2.5 mg / kg), 6a (5 mg / kg), and 6a (10 mg / kg) can significantly increase the amount of CA induced by aconitine in rats (P < 0.05). Compared with nuciferine, 6a (5 mg / kg) and 6a (10 mg / kg) can significantly increase the amount of VT induced by aconitine in rats (P < 0.05). Compared with nuciferine, 4 (5 mg / kg), 6a (2.5 mg / kg), 6a (5 mg / kg) and 6a (10 mg / kg) can significantly increase the amount of VF induced by aconitine in rats (P < 0.05). Compared with nuciferine, 6a (5 mg / kg) and 6a (10 mg / kg) can significantly increase the amount of CA induced by aconitine in rats (P < 0.01).

[0225] Example 22 Therapeutic effect of the compound represented by formula 6a on BaCl2-induced arrhythmia in rats

[0226] Through the effect of nuciferine derivatives on BaCl2-induced arrhythmia in rats, the present invention screened out the compound 6a with the best efficacy to explore the effect of 6a on BaCl2-induced arrhythmia in rats at different concentrations. The experimental results are as follows Fig.40 , Fig.41 As shown, compared with normal saline and DMSO, different concentrations of 6a significantly reduced the time for the recovery of normal heart rate after BaCl2-induced arrhythmia in rats (P < 0.001). Compared with DMSO, 2.5 mg / kg of 6a significantly increased the maintenance time after the recovery of BaCl2-induced arrhythmia (P < 0.05). Compared with normal saline and DMSO, 5 mg / kg and 10 mg / kg of 6a significantly increased the maintenance time after the recovery of BaCl2-induced arrhythmia (P < 0.001). Compared with nuciferine and verapamil, 5 mg / kg and 10 mg / kg of 6a showed a significant trend of increasing the maintenance time after the recovery of BaCl2-induced arrhythmia. From the above experimental results, it can be seen that within a certain concentration range, the therapeutic effect of 6a on BaCl2-induced arrhythmia in rats will gradually increase with the increase of 6a concentration.

[0227] Example 23 Effect of the compound represented by formula 6a on the heart rate of normal rats

[0228] The SD rats were randomly divided into groups and connected to the BL-420N biological signal acquisition system after anesthesia. The normal ECG was recorded for 1 minute. After the sublingual intravenous injection of the nuciferine derivative 6a (5 mg / kg) at 60 seconds, the ECG was observed for 35 minutes. The electrocardiogram (HR, QT, RR, QTc, R voltage) was recorded at 30 seconds (normal), 90 seconds, 3 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, and 35 minutes. Fig.42 As shown, 6a after injection, the heart rate of rats decreased significantly (P < 0.05), and the RR interval time increased significantly (P < 0.05) compared with 30s (normal). The other electrocardiogram indices (QT, QTc, R voltage) showed no significant differences compared with the normal group (P > 0.05).

[0229] Example 24 Effects of the compound represented by formula 6a on electrophysiological signals of rat ventricular myocytes

[0230] The voltage-dependent calcium current (I Ca ), sodium current (I Na ) and action potential duration (APD) as well as measurement of hERG.

[0231] Patch clamp data recording:

[0232] Whole-cell patch clamp experiments were performed in HEK293 cells in voltage clamp mode using an EPC-10 amplifier and patchmaster software (HEKA Elektronik, Germany) and analyzed by Clampfit 10.6.

[0233] For APD recording, the cell culture medium consisted of 137 mM NaCl, 4 mM KCl, 10 mM HEPES, 10 mM D-glucose, 1 mM MgCl2, and 2 mM CaCl2, and the pH was adjusted to 7.3 using NaOH. The solution in the glass electrode consisted of 4 mM NaCl, 1 mM MgCl2, 0.1 mM CaCl2, 138 mM K-gluconate, 10 mM HEPES, 1 mM EGTA, and 2 mM Mg-ATP, and the pH was adjusted to 7.3 using KOH.

[0234] For Nav1.5 recording, the cell culture medium consisted of 137mM NaCl, 10mM D-glucose, 4mM KCl, 1.8mM CaCl2, 1mM MgCl2 and 10mM HEPES, and the pH was adjusted to 7.4 with NaOH. The solution in the glass electrode: 50mM CsCl, 10mM NaCl, 10mM HEPES, 60mM CsF and 20mM EGTA, and the pH was adjusted to 7.2 with CsOH.

[0235] For Cav1.2 recordings, the solution in the glass electrode consisted of 110 mM CsCl, 10 mM EGTA, 5 mM HEPES, 1 mM CaCl2, 4 mM Na2-ATP and 4.5 mM Mg-ATP, and the pH was adjusted to 7.2 with CsOH; the cell culture medium consisted of 140 mM TEA-Cl, 1 mM MgCl2, 4 mM KCl, 10 mM CaCl2, 5 mM D-glucose, and 10 mM HEPES, and the pH was adjusted to 7.4 with TEA-OH.

[0236] For hERG recordings, the cell culture medium was the same as that used for recording APD from ventricular myocytes. The solution in the glass electrode consisted of 20 mM KCl, 120 mM K-aspartate, 10 mM HEPES, 10 mM EGTA, 5 mM MgCl2, and 5 mM Mg-ATP, adjusted to pH 7.2 with KOH.

[0237] Example 24-1 Effect of the compound represented by formula 6a on the action potential duration (APD) of rat cardiomyocytes

[0238] like Fig.43 As shown, compound 6a (10 μM) can significantly reduce the amplitude of action potential (p < 0.001) and resting potential (p < 0.05) in current clamp mode. At the same time, it has no effect on APD30 and APD50 of rat ventricular myocytes, but can prolong APD90, which indicates that compound 6a can prolong ventricular conduction in rats and mildly block sodium ion influx, thereby slightly reducing APA and resting potential.

[0239] Example 24-2 Effect of the compound represented by formula 6a on Nav and Cav currents in rat myocardial cells

[0240] Example 24-2-1 Effect of the compound represented by formula 6a on the Nav current of rat myocardial cells

[0241] like Fig.44 As shown, in the whole-cell patch clamp current clamp mode, the compound represented by formula 6a (10 μM) can significantly block the INa current, and does not change the current-voltage relationship of the activation potential and peak potential ( Fig.44 A). Fig.44 The BF in Figure 6 shows that the compound (10 μM) of Formula 6a does not affect the recovery of I in a voltage-dependent manner. Na Steady-state activation of I Na Steady-state inactivation and I Na This result also shows that the compound represented by formula 6a (10 μM) can change I Na The activation, inactivation and recovery characteristics of the channel. Fig.44 The current-voltage curve in B shows that 6a can significantly increase I Na The IV curve of the activation curve did not shift significantly to the right, and the V 1 / 2 The inactivation curve shifted to the right, V 1 / 2 The value of k shifted from (-84.45±7.360)mV to (-103.4±3.458)mV (n=13, P<0.001), and the value of k changed from (-5.076±0.9617)mV to (-4.210±0.8139)mV (n=13, P<0.05), indicating that the compound represented by Formula 6a binds to the Nav channel with strong affinity and accelerates the Nav inactivation process. After treatment with the compound shown in formula 6a, the recovery curve shifted to the right, the τ value changed from 7.255±0.7848 to 34.23±7.541 (n=13, P<0.01), and the k value changed from 0.1582±0.01723 to 0.04124±0.007670 (n=13, P<0.001), indicating that compound 6a can significantly change the Nav recovery kinetics after inactivation and delay the resurrection process of the sodium current channel. The above results show that compound 6a has a strong affinity in the process from inactivation to activation of the Nav channel and can connect well with the Nav channel.

[0242] Example 24-2-2 Effect of the compound represented by formula 6a on Cav current of rat ventricular myocytes

[0243] The results of the whole-cell patch clamp in current clamp mode showed that the compound represented by formula 6a (10 μM) did not change the I Ca Under the condition of the current-voltage curve trend, it can significantly block the I Ca Current (****p<0.001) ( Fig.45A). Fig.45 The BF in Figure 1 shows the I Ca Homeostatic activation, I Ca Steady-state inactivation and I Ca The recovery curve showed that the compound represented by formula 6a (10 μM) could change the I Ca Channel. Fig.45 The current-voltage curve in B shows that 6a can significantly increase I Ca The IV curve of the activation curve remains unchanged, V 1 / 2 The value of kinetics of the ... ca The inactivation curve of V 1 / 2 From (-3.343±3.817)mV to (-12.860±3.633)mV (n=7, P<0.001), k from (-4.025±1.065)mV to (-6.344±1.507)mV (n=7, P<0.05), indicating that compound 6a accelerated the inactivation process of Cav. After treatment with compound 6a, the recovery curve shifted positively along the time axis, the τ value changed from 0.08245±0.009048 to 0.1921±0.03795 (n=7, P<0.05), and the k value changed from 13.16±1.618 to 6.414±1.141 (n=7, P<0.01), indicating that compound 6a can reasonably change the recovery kinetics of Cav after inactivation and moderately delay the resurrection process of calcium current channels. This also means that compound 6a has a strong affinity in the process from inactivation to activation of Cav channels.

[0244] Example 24-2-3 Effect of the compound represented by formula 6a on rat ventricular myocytes Nav1.5

[0245] In order to provide compound 6a for the inward current I Na In order to understand the mechanism of the effect, the present invention studied the effect of compound 6a on Nav1.5 channels in HEK293 cells. Fig.46 As shown, compound 6a exhibited a concentration-dependent inhibitory effect on sodium current in Nav1.5 channels of HEK293. The IC in the resting state was calculated based on the dose-response curve. 50 The value is 23.886μM. In the half-inactive state, IC 50 The value is 4.9857 μM.

[0246] Example 24-2-4 Effect of the compound represented by formula 6a on rat cardiomyocyte Cav1.2

[0247] like Fig.47 As shown, compound 6a has a concentration-dependent inhibitory effect on Cav1.2 current in HEK293 cells. As the concentration of 6a gradually increased, I Ca Concentration response curve of peak inhibition. The inhibitory effect of compound 6a on Cav1.2 channels is usually determined by the state of Cav1.2 channels. In the half-inactivated state, the IC calculated from the concentration response curve is 50 The value is 4.6228μM, Fig.47 It can be seen that the voltage decreased after washing out 6a, indicating that the binding of 6a to Cav1.2 is partially reversible.

[0248] Example 24-2-5 Effect of the compound represented by formula 6a on rat myocardial cell hERG / IKr

[0249] If the inhibitory effect of the drug on hERG / IKr is too strong, it will cause a sharp prolongation of the QT interval and cause long QT syndrome. The inhibitory effect of the drug on hERG / IKr is usually determined by IC 50 To assess, when IC 50 <1μM is high risk, 1μM<IC 50 <10μM is medium risk, IC 50 >10μM is low risk, IC 50 >30μM is very low risk. In general, IC 50 The lower it is, the greater the risk of developing long QT syndrome. Fig.48 It can be seen that compound 6a inhibits Kv11.1 channels in a concentration-dependent manner, thereby blocking hERG currents. The IC of 6a for Kv11.1 channel current inhibition is 50 The inhibitory effect was reversible. Fig.49 As shown, the IC50 calculation result of the positive control drug cisapride is 13.914nΜ. Compared with the positive drug, the IC50 of 6a on Kv11.1 channel current is 50 Higher, from the above results, it can be concluded that the risk of 6a inducing long QT syndrome is extremely low.

[0250] According to all the experimental results above, all the synthesized nuciferine derivatives have different degrees of anti-arrhythmic effects, and the compound represented by formula 6a, which has the best activity, is better than verapamil in the treatment of arrhythmias caused by barium chloride or aconitine. In the whole-cell patch clamp, it was shown that the compound represented by formula 6a can bind to Nav1.5 and Cav1.2 and inhibit the protein activity of Nav1.5 and Cav1.2, causing I Na and I Ca Reduce; although the compound shown in Formula 6a can also bind to Kv11.1, its inhibition rate on Kv11.1 is lower than that of the positive drug cisapride, and the risk of causing long QT syndrome is lower. From the above results, it can be seen that the compound shown in Formula 6a has the characteristics of good activity and low toxicity for the treatment of arrhythmias.

[0251] In summary, the present invention provides a series of nuciferine derivatives for use in antiarrhythmics through total synthesis. Experimental results show that the compounds provided by the present invention have excellent antiarrhythmic effects. The compound shown in Formula 6a has a stronger therapeutic effect on arrhythmias than the positive drug verapamil and a lower probability of causing adverse reactions (long QT syndrome). Therefore, the compounds provided by the present invention provide a new option for preparing low-toxic and highly active antiarrhythmic drugs.

[0252] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A nuciferine derivative, characterized in that: The structure of the nuciferine derivative is shown below: The R group structure is any of the following structures:

2. The method for preparing the nuciferine derivative according to claim 1, characterized in that: The chemical reaction formula is as follows:

3. The preparation method according to claim 2, characterized in that: The preparation method of the compound 2-(2-bromophenyl)-N-(3,4-dimethoxyphenethyl)acetamide shown in Formula 1 is as follows: 2-bromophenylacetic acid is dissolved in dichloromethane, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3,4-dimethoxyphenethylamine and triethylamine are added and reacted to obtain the compound shown in Formula 1; The preparation method of the compound 1-(2-bromobenzyl)-6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline shown in Formula 2 is as follows: dissolving the compound shown in Formula 1 with DCM, adding phosphorus pentachloride, refluxing the reaction, drying the DCM, adding MeOH and sodium borohydride, and reacting to obtain the compound shown in Formula 2; The preparation method of the compound 1-(1-(2-bromobenzyl)-6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl)-2,2,2-trifluoroethane-1-one represented by formula 3 is as follows: dissolving the compound represented by formula 2 with DCM, adding trifluoroacetic anhydride and Et3N, and reacting to obtain the compound represented by formula 3; The preparation method of the compound 1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)-2,2,2-trifluoroethane-1-one shown in formula 4 is as follows: dissolving the compound shown in formula 3 with N,N-dimethylacetamide, adding tri(4-fluorophenyl)phosphine, K2CO3 and palladium acetate to react to obtain the compound shown in formula 4; The preparation method of the compound 1,2-dimethoxy-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline represented by Formula 5 is as follows: dissolving the compound represented by Formula 4 with DCM, adding NaBH4, and reacting to obtain the compound represented by Formula 5.

4. The preparation method according to claim 2, characterized in that: When the R group structure is 6a, the preparation method of the compound 2-(2-bromophenyl)-1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)ethan-1-one represented by formula 6a is as follows: dissolving the compound represented by formula 5 with DMF, adding 2-bromophenylacetic acid, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine, and reacting to obtain the compound represented by formula 6a; When the R group structure is 6b, the preparation method of the compound (4-chloromethylphenyl)(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)methanone represented by formula 6b is as follows: dissolving the compound represented by formula 5 with DMF, adding 4-(chloromethyl)benzoic acid, HATU and DIEPA, and reacting to obtain the compound represented by formula 6b; When the R group structure is 6c, the preparation method of the compound (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(4-fluorophenyl)methanone represented by formula 6c is as follows: dissolving the compound represented by formula 5 with DMF, adding 4-fluorobenzoic acid, HATU and DIEPA, and reacting to obtain the compound represented by formula 6c; When the R group structure is 6d, the synthesis of the compound represented by formula 6d (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(4-methoxyphenyl)methanone: dissolve the compound represented by formula 5 in DMF, add 4-methoxybenzoic acid, HATU and DIEPA, and react to obtain the compound represented by formula 6d; When the R group structure is 6g, the synthesis of the compound 1-(1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)prop-2-en-1-one shown in formula 6g: dissolve the compound shown in formula 5 in DMF, add acrylic acid, HATU and DIEPA, and react to obtain the compound shown in formula 6g; When the R group structure is 6h, the synthesis of the compound represented by formula 6h (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(2-methylfuran-3-yl)methanone: dissolve the compound represented by formula 5 in DMF, add 2-methylfuran-3-carboxylic acid, HATU and DIEPA, and react to obtain the compound represented by formula 6h; When the R group structure is 6e, the compound represented by Formula 6e (1,2-dimethoxy-6-toluenesulfonyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline) is synthesized by dissolving the compound represented by Formula 5 in DCM, adding p-toluenesulfonyl chloride and Et3N to react to obtain the compound represented by Formula 6e; When the R group structure is 6f, the synthesis of the compound represented by formula 6f (1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(pyrrolidin-1-yl)methanone: dissolving the compound represented by formula 5 in DMF, adding pyrrolidine-1-carbonyl chloride, potassium carbonate and potassium iodide, and reacting to obtain the compound represented by formula 6f; When the R group structure is 6i, the synthesis of the compound 1,2-dimethoxy-6-(methylsulfonyl)-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline represented by formula 6i: dissolve the compound represented by formula 5 in DCM, add methylsulfonyl chloride and Et3N, and react to obtain the compound represented by formula 6i; When the R group structure is 6j, the synthesis of the compound represented by formula 6j ((1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(4-methylpiperazine-1-yl)methanone) is as follows: the compound represented by formula 5 is dissolved in DCM, 4-methylpiperazine-1-carbonyl chloride, K2CO3 and KI are added and reacted to obtain the compound represented by formula 6j; When the R group structure is 61, the synthesis of the compound shown in formula 61 ((1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinolin-6-yl)(morpholinyl)methanone) is as follows: the compound shown in formula 5 is dissolved in DCM, and morpholine-4-carbonyl chloride, K2CO3 and KI are added and reacted to obtain the compound shown in formula 61; When the R group structure is 6m, the synthesis of the compound represented by formula 6m (N,N-diethyl-1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-carboxamide) is as follows: the compound represented by formula 5 is dissolved in DCM, and diethylcarbamoyl chloride, K2CO3 and KI are added and reacted to obtain the compound represented by formula 6m; When the R group structure is 6n, the synthesis of the compound represented by formula 6n (N,N-dimethyl-1,2-dimethoxy-4,5,6a,7-tetrahydro-6H-dibenzo[de,g]quinoline-6-carboxamide) is as follows: the compound represented by formula 5 is dissolved in DCM, and dimethylcarbamoyl chloride, K2CO3 and KI are added and reacted to obtain the compound represented by formula 6n; When the R group structure is 6k, the synthesis of the compound represented by formula 6k (1,2-dimethoxy-6-(4-methoxybenzyl)-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline): the compound represented by formula 5 is dissolved in DCM, and p-methoxybenzaldehyde and sodium triacetylborohydride are added and reacted to obtain the compound represented by formula 6k.

5. Use of the nuciferine derivative prepared by the preparation method according to any one of claims 2 to 4 in the preparation of drugs for treating arrhythmia.

6. The use according to claim 5, characterized in that: The arrhythmia includes sinus arrhythmia, atrial arrhythmia, supraventricular arrhythmia and ventricular arrhythmia.

7. The use according to claim 6, characterized in that: The sinus arrhythmia includes sinus bradycardia, sinus tachycardia, sinus arrhythmia, sinus arrest and sinoatrial block; the atrial arrhythmia includes atrial premature beats, atrial tachycardia, and atrial premature contractions; the supraventricular arrhythmia includes paroxysmal supraventricular tachycardia; the ventricular arrhythmia includes ventricular premature beats, ventricular tachycardia, and ventricular fibrillation.

8. The use according to claim 5, characterized in that: The medicine may also include a pharmaceutically acceptable carrier; the pharmaceutically acceptable carrier is selected from one or more of excipients, buffers, emulsifiers, stabilizers, diluents, adhesives, preservatives, lubricants, pH regulators, cryoprotectants, and flavoring agents.

9. The use according to claim 5, characterized in that: The dosage form of the drug is drops, mixtures, tinctures, injections, tablets, powders, oral liquids, capsules, granules, ointments, suspensions, powders, emulsions, solutions, pellets, pills, buccal tablets, freeze-dried powder injections, gels, suppositories or aerosols.

10. Use of the nuciferine derivative prepared by the preparation method according to any one of claims 2 to 4 in the preparation of a drug for treating QT syndrome.

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