Isoquinoline alkaloid compounds from picraline and preparation and application and composition thereof

Abstract

The present application relates to a kind of new isoquinoline alkaloid compound and its preparation method and purposes.The new compound (Corybungine A-L) provided by the present application is extracted and separated from Papaveraceae Corydalis bungeana Turcz., a kind of novel isoquinoline alkaloid with novel structure.Biological activity experiment shows that the alkaloid compound has dopamine D2 receptor antagonistic activity, and can be applied in the preparation of dopamine D2 receptor related pain, schizophrenia, Lesch-Nyhan syndrome and other nervous system diseases.

Description

[0001] The present application is a divisional application of a Chinese invention patent application with the original application date of November 30, 2021, the application number of 202111441010.5, the title of Corybungine A-L and the preparation and application thereof and the composition, the publication number of CN116199694A; due to the original application having the problem of single pointed out by the examiner, the applicant proposes a divisional application. TECHNICAL FIELD

[0002] The present application belongs to the field of natural medicinal chemistry, and relates to a new isoquinoline alkaloid compound. More specifically, it relates to a new isoquinoline compound (Corybungine A-L) and a preparation method thereof and the use thereof as a dopamine D2 receptor active ingredient for treating diseases related to the nervous system, such as pain, schizophrenia, Lesch-Nyhan syndrome, and the like. In vitro activity shows that the above-mentioned compound has significant dopamine D2 receptor antagonistic activity, and can be used for developing new drugs or lead compounds for treating or inhibiting diseases related to the nervous system. BACKGROUND

[0003] Screening of lead compounds with pharmacological activity from natural products is an important way for drug research and development. Natural products have the characteristics of structural complexity and structural diversity, and the toxic and side effects are often small, which is a good class of compounds for drug development. According to statistics, among the 1881 new drugs listed from 1981 to 2019, about 50% are directly or indirectly derived from natural products. According to literature reports, among the natural products with pharmacological activity, alkaloids account for about 50%. Systematic separation and purification of such alkaloid-rich medicinal plants is conducive to the discovery of more potential high-activity alkaloid compounds, and provides a guide for the discovery of lead compounds and the development of new drugs.

[0004] Corydalis bungeana Turcz. is a traditional Chinese and Mongolian medicine material, which is the dried whole herb with roots of Corydalis bungeana Turcz. of the family Papaveraceae. Corydalis bungeana Turcz. is bitter and cold in nature, and is often used for clearing heat and resolving toxins, resolving masses and relieving swelling, and treating various diseases such as cold and cough, and has good effects on rheumatism and myocarditis. According to literature reports, Corydalis bungeana Turcz. contains flavonoids and alkaloids. Alkaloids are the most common chemical components in Corydalis bungeana Turcz., and so far, more than thirty alkaloids have been found in Corydalis bungeana Turcz., including corybulbine, acetyl corybulbine, protopine, dihydrosanguinarine, and the like. Modern pharmacological studies have shown that Corydalis bungeana Turcz. has various pharmacological effects such as analgesic and anti-inflammatory, sedative and hypnotic, antiviral, antibacterial, immunosuppressive, and cytotoxic effects.

[0005] Dopamine receptors are a class of G protein-coupled receptors. Currently discovered dopamine receptors can be mainly divided into two categories: D1 class receptors mainly including D1 and D5 receptors; and D2 class receptors mainly including D2, D3 and D4 receptors. G protein-coupled dopamine receptors (D1, D2, D3, D4 and D5) mediate all physiological functions of catecholamine neurotransmitter dopamine, including voluntary movement and excitement to regulating hormones and hypertension. Dopamine D2 receptor antagonistic activity has been reported to be related to many nervous system diseases, such as pain, schizophrenia, Lesch-Nyhan syndrome and the like. Modern pharmacological research shows that kudou-ding has analgesic anti-inflammatory, sedative and hypnotic effects, and therefore, it is of important research value to find compounds with dopamine D2 receptor activity from kudou-ding.

[0006] The six aporphine isoquinoline compounds (Corybungine A-F), one protoberberine isoquinoline compound (Corybungine G) and five open-ring protoberberine isoquinoline compounds (Corybungine H-L) in the present application are new compounds which are extracted and separated from kudou-ding for the first time, and have not been reported in other natural sources, and there is no report about the compounds and their pharmacological activities in the existing literatures. The pharmacological activity test shows that the compounds have significant dopamine D2 receptor antagonistic activity, and can be applied in the preparation of medicines for treating diseases such as pain, schizophrenia, Lesch-Nyhan syndrome and the like. SUMMARY

[0007] The present application provides a novel isoquinoline compound, or different crystal forms of the compound, or chiral isomers thereof, or glycosides thereof, or pharmaceutically acceptable salts thereof, or solvates thereof, or prodrugs thereof, or metabolites thereof, and the structural general formula (I) thereof is as follows:

[0008]

[0009] wherein R1-R 14 are each independently hydrogen, chlorine, hydroxyl, methoxyl, ethoxyl, O-glycosyl or OAr; the Ar is substituted or unsubstituted phenyl; and the glycosyl is monosaccharide glycosyl or various disaccharide glycosyl and polysaccharide glycosyl formed by monosaccharide.

[0010] Further, the stereochemical configuration of the compounds 1-6 is selected from one or two of 6aR and 6aS, and the stereochemical configuration of the compound 7 is selected from one or two of 14R and 14S; R1-R 14In the above-mentioned formula, Ar is a substituted or unsubstituted phenyl; the substituted phenyl refers to a phenyl group optionally substituted with one or more substituents, wherein each substituent can be independently a hydroxyl group, a hydroxymethyl group, a methoxyl group or OAr'; Ar' is a substituted or unsubstituted phenyl; the substituted phenyl Ar' refers to a phenyl group optionally substituted with one or more substituents, wherein each substituent can be independently a hydroxyl group, a hydroxymethyl group, a methoxyl group or an ethoxyl group. 14 In the above-mentioned formula, the sugar group refers to, but is not limited to, a glucosyl group, a glucuronosyl group, a mannosyl group, a galactosyl group, an allosyl group, a fructosyl group, a sorbosyl group, a fucosyl group, a rhamnosyl group, a cinamoyl group, an arabinosyl group, a lyxosyl group, a xylosyl group, a ribosyl group, and various disaccharide groups and polysaccharide groups formed by the above-mentioned monosaccharides.

[0011]

[0012] Further, the stereo configuration of compounds 1-4 and 6 is preferably 6aR, respectively, the stereo configuration of compound 5 is preferably 6aS, and the stereo configuration of compound 7 is preferably 14S; compounds 1-12 have the structure in the following formula (II), and are named as Corybungine A-L in sequence, respectively.

[0013] The present application also provides a method for preparing the above-mentioned compound (II), which comprises the following steps: grinding dry whole plants of Kudou Ding to obtain medicinal material powder, extracting the medicinal material powder with 50-90% (by volume) ethanol at 60-70°C for 6-24 hours, filtering the extract with a 80-mesh filter screen, and concentrating the extract under reduced pressure to obtain extract powder; dissolving the extract powder in 0.01-0.2 mol / L sulfuric acid, adjusting the pH to 2-3, adding petroleum ether in a volume ratio of 1:1-1:2 to the solution, and taking the water layer; adjusting the pH of the obtained water layer to 9-10 with 0.01-2 mol / L sodium hydroxide, adding dichloromethane in a volume ratio of 1:1-1:2 to the solution, and taking the dichloromethane layer; and separating the dichloromethane layer through multi-step high-performance preparative liquid chromatography to obtain compounds Corybungine A-L.

[0014] The obtained isoquinoline alkaloid compound is subjected to activity test of dopamine D2 receptor, and the activity test is performed by using Chinese hamster ovary cells CHO transfected with dopamine D2 receptor, and the results show that the compound exhibits low micromolar antagonistic activity on dopamine D2 receptor, and can become a potential lead compound for treating nervous system diseases such as pain, schizophrenia, Lesch-Nyhan syndrome and the like.

[0015] In the present application, the sugar group refers to, but is not limited to, a glucosyl group, a glucuronyl group, a mannosyl group, a galactosyl group, an allosyl group, a fructosyl group, a sorbosyl group, a fucosyl group, a rhamnosyl group, a cineroyl group, an arabinosyl group, a lyxosyl group, a xylosyl group, a ribosyl group, and various disaccharide groups and polysaccharide groups formed by the above-mentioned monosaccharides.

[0016] The compound of the present application can be obtained by purification from plants or synthesized by chemical methods well known to those skilled in the art.

[0017] The compound of the present application can be used alone or in combination, and can be combined with a pharmaceutically suitable carrier or excipient to form an oral or non-oral dosage form according to conventional methods.

[0018] Obviously, according to the above content of the present application, other various forms of modification, replacement or change can be made without departing from the above-mentioned basic technical idea of the present application according to the ordinary technical knowledge and common means in the art.

[0019] The novel compound (Corybungine A-L) provided by the present application is a novel isoquinoline alkaloid extracted and separated from Corydalis bungeana Turcz. of the Corydalis genus of the Papaveraceae family. Bioactivity experiments show that the alkaloid compound has dopamine D2 receptor antagonistic activity and can be applied to the preparation of drugs for dopamine D2 receptor related nervous system diseases such as pain, schizophrenia, Lesch-Nyhan syndrome, etc.

[0020] The present application has the following advantages: the target compound is a novel isoquinoline alkaloid compound; the compound has significant dopamine D2 receptor antagonistic activity and has application prospects for the development of drugs for currently highly concerned central nervous system diseases such as pain, schizophrenia, Lesch-Nyhan syndrome, etc. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Compound 1-12 1 H NMR and 13 C NMR spectrum, wherein a is the H NMR and 1 H NMR and 13 C NMR spectrum, b is the H NMR and 1 H NMR and 13 C NMR spectrum, c is the H NMR and 1 H NMR and 13 C NMR spectrum, d is the H NMR and 1 H NMR and 13 C NMR spectrum, e is the H NMR and1 HNMR and 13 CNMR spectrum, f is compound 6 1 HNMR and 13 CNMR spectrum, g is compound 7 1 HNMR and 13 CNMR spectrum, h is compound 8 1 HNMR and 13 CNMR spectrum, i is compound 9 1 HNMR and 13 CNMR spectrum, j is compound 10 1 HNMR and 13 CNMR spectrum, k is compound 11 1 HNMR and 13 CNMR spectrum, 1 is compound 12 1 HNMR and 13 CNMR spectrum;

[0022] Figure 2 Characterization of the antagonistic activity of compounds 1-12 on D2 receptors; 1 H, 1 H-COSY and key HMBC;

[0023] Figure 3 Characterization of the antagonistic activity of compounds 1, 3, 8 on D2 receptors;

[0024] Figure 4 Structure of compounds 1-12, which are preferred embodiments of the application. DETAILED DESCRIPTION

[0025] The following examples are intended to illustrate the application and not to further limit it, the application can be practiced in any way described in the summary.

[0026] Example of preparation of compounds of formula (II) of the application:

[0027] Preparation of compounds and structure identification:

[0028] In the following preparation examples, the preparation system includes Waters Alliance, including e2695 separation unit, 2998 PDA detector, data processing by empower 3; Waters AutoP automatic purification system, including 2545 separation unit, 2767 sample manager, 2489 dual wavelength detector; NovaSep HPLC industrial high pressure liquid preparation chromatography and HIPERSEP software. Reagents include chromatographic grade methanol and acetonitrile purchased from Fisher Scientific (Loughborough, UK), chromatographic grade formic acid, acetic acid, ammonia, triethylamine, ammonium formate, sodium monobasic phosphate, sodium dihydrogen phosphate purchased from Biotang (Hebei, China), laboratory water from Milli-Q ultrapure water purification system (Billerica, MA, USA), preparation grade methanol purchased from Shanghai Xingke High Purity Solvent Co., Ltd. (Shanghai, China). Preparation column and semi-preparation column: C18HCE (15 μm, 100 mm x 250 mm), C18HCE (10 μm, 100 mm x 325 mm), C18CE (7 μm, 50 mm x 250 mm), FC8HL (3.5 μm, 30 mm x 150 mm) and C18HCE (5 μm, 10 mm x 150 mm) (Dalian Sipu Jinggong Co., Ltd.).

[0029] The nuclear magnetic resonance spectrum used for structure identification was determined by Bruker AVIII-600 nuclear magnetic resonance spectrometer (Bruker, German), and the compounds were dissolved in deuterated methanol (MeOD). Mass spectrometry was performed by Agilent 1290 Infinity LC / 6540 Q-TOF MS liquid chromatography-mass spectrometry system for sample separation and analysis.

[0030] The preparation and compound identification steps are as follows:

[0031] (1) Extraction of medicinal materials: 100 kg of dry root whole grass of Ixeridium compositum was obtained by crushing. 1000 liters of 70% ethanol by volume was heated to 70°C for 6 hours, and the extract was filtered through an 80 mesh filter. The solvent was removed by rotary evaporation to obtain the extract. The extract was dissolved in 0.01 mol / L sulfuric acid, adjusted to pH 2, and extracted with petroleum ether at a volume ratio of 1:1. The water layer was obtained. The obtained water layer was adjusted to pH 10 with 2 mol / L sodium hydroxide, and extracted with dichloromethane at a volume ratio of 1:1. The dichloromethane layer obtained was the crude base.

[0032] (2) The crude base obtained in step (1) was subjected to first dimensional separation and purification using C18HCE (15um, 100x250mm, reversed-phase column) at a flow rate of 320 mL / min, mobile phase A being (0.1% by volume concentration) formic acid-methanol and B being (0.1% by volume concentration) formic acid-water, with gradient elution conditions being: 0-8 min: 25% A (by volume ratio, the same below), 8-23 min: 40% A, 23-38 min: 80% A, 38-55 min: 100% A, and samples being taken at 4.2-26.0 min, and after concentration, the total alkaloids of Corydalis Bungeana were obtained.

[0033] (3) The total alkaloids of Corydalis Bungeana obtained in step (2) were subjected to second dimensional separation and purification using C18HCE (10um, 100x325mm, reversed-phase column) at a flow rate of 320 mL / min, mobile phase A being (0.1% by volume concentration) formic acid-methanol and B being (0.1% by volume concentration) formic acid-water, with gradient elution conditions being: 0-5 min: 20% A, 5-50 min: 20-45% A, 50-60 min: 95% A, and 11 sub-fractions F1-F11 were obtained by collection according to chromatographic peaks.

[0034] (4) Sub-fraction F2 obtained in step (3) was subjected to third dimensional separation and purification using C8CE (reversed-phase column) at a flow rate of 320 mL / min, mobile phase A being (0.1% by volume concentration) 25% ammonia water-methanol and B being (0.1% by volume concentration) 25% ammonia water-water, with gradient elution conditions being: 0-5 min: 5% A, 5-50 min: 5-95% A, 50-60 min: 95% A, and 14 sub-fractions F2-1-F2-14 were obtained by collection according to chromatographic peaks.

[0035] (5) Sub-fraction F2-10 obtained in step (4) was subjected to fourth dimensional separation and purification using FC8HL (reversed-phase column) at a flow rate of 320 mL / min, mobile phase A being (20 mM) ammonium formate-95% methanol / water (by volume concentration) and B being (20 mM) ammonium formate-water, with gradient elution conditions being: 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, 13.88-32 min: 100% A, and 5 sub-fractions F2-10-1-F2-10-5 were obtained by collection according to chromatographic peaks.

[0036] (6) Sub-fraction F2-10-1 obtained in step (5) was subjected to fifth dimensional separation and purification using C18HCE (reversed-phase column) at a flow rate of 320 mL / min, mobile phase A being (0.1% by volume concentration) formic acid-methanol and B being (0.1% by volume concentration) formic acid-water, with gradient elution conditions being: 0-8 min: 40% A, 8-20 min: 55% A, and compound 4 (t R = 8.0 min) was obtained by collection according to chromatographic peaks, and was named Corybungine D.

[0037] (7) The sub-fraction F3 obtained in step (3) was subjected to third dimensional separation and purification using C8CE (reversed-phase chromatographic column) with mobile phase A of (0.1% by volume) 25% ammonia water-methanol and B of (0.1% by volume) 25% ammonia water-water, and gradient elution conditions of 0-5 min: 5% A, 5-50 min: 5-95% A, and 50-60 min: 95% A, to obtain 12 sub-fractions F3-1 to F3-12 by peak collection.

[0038] (8) The sub-fraction F3-12 obtained in step (7) was subjected to fourth dimensional separation and purification using FC8HL (reversed-phase chromatographic column) with mobile phase A of (20 mM) ammonium formate-95% methanol / water (by volume) and B of (20 mM) ammonium formate-water, and gradient elution conditions of 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, and 13.88-32 min: 100% A, to obtain 8 sub-fractions F3-12-1 to F3-12-8 by peak collection.

[0039] (9) The sub-fraction F3-12-5 obtained in step (8) was subjected to fifth dimensional separation and purification using C18HCE (reversed-phase chromatographic column) with mobile phase A of (0.1% by volume) formic acid-methanol and B of (0.1% by volume) formic acid-water, and gradient elution conditions of 0-15 min: 29% A, to obtain compound 5 (t R = 8.5 min), named as Corybungine E.

[0040] (10) The sub-fraction F4 obtained in step (3) was subjected to third dimensional separation and purification using C8CE (reversed-phase chromatographic column) with mobile phase A of (0.1% by volume) 25% ammonia water-methanol and B of (0.1% by volume) 25% ammonia water-water, and gradient elution conditions of 0-5 min: 5% A, 5-50 min: 5-95% A, and 50-60 min: 95% A, to obtain 15 sub-fractions F4-1 to F4-15 by peak collection.

[0041] (11) The sub-fraction F4-13 obtained in step (10) was subjected to fourth dimensional separation and purification using FC8HL (reversed-phase chromatographic column) with mobile phase A of (20 mM) ammonium formate-95% methanol / water (by volume) and B of (20 mM) ammonium formate-water, and gradient elution conditions of 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, and 13.88-32 min: 100% A, to obtain 10 sub-fractions F4-13-1 to F4-13-10 by peak collection.

[0042] (12) The sub-fraction F4-13-4 obtained in step (11) was subjected to fifth dimensional separation and purification by C18HCE (reversed-phase column) with mobile phase A of (0.1% v / v) formic acid-methanol and B of (0.1% v / v) formic acid-water under gradient elution conditions of 0-5 min: 25% A, 5-15 min: 25-45% A, 15-20 min: 45% A, and 18 compounds 3 (tR= 10.0 min), 7 (tR= 15.4 min), 8 (tR= 13.6 min), 9 (tR= 18.1 min) were collected as chromatographic peaks and named as Corybungine C, G, H and I, respectively. R R R R

[0043] (13) The sub-fraction F4-13-7 obtained in step (11) was subjected to fifth dimensional separation and purification by C18HCE (reversed-phase column) with mobile phase A of (0.1% v / v) formic acid-methanol and B of (0.1% v / v) formic acid-water under gradient elution conditions of 0-12 min: 25% A, 12-15 min: 25-45% A, 15-20 min: 45% A, and 2 compounds 1 (tR= 18.1 min) and 2 (tR= 18.9 min) were collected as chromatographic peaks and named as Corybungine A and B, respectively. R R

[0044] (14) The sub-fraction F6 obtained in step (3) was subjected to third dimensional separation and purification by C8CE (reversed-phase column) with mobile phase A of (0.1% v / v) 25% ammonia-methanol and B of (0.1% v / v) 25% ammonia-water under gradient elution conditions of 0-5 min: 5% A, 5-50 min: 5-95% A, 50-60 min: 95% A, and 18 sub-fractions F6-1 to F6-18 were collected as chromatographic peaks.

[0045] (15) The sub-fraction F6-8 obtained in step (14) was subjected to fourth dimensional separation and purification by FC8HL (reversed-phase column) with mobile phase A of (20 mM) ammonium formate-95% methanol / water (v / v) and B of (20 mM) ammonium formate-water under gradient elution conditions of 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, 13.88-32 min: 100% A, and 15 sub-fractions F6-8-1 to F6-8-15 were collected as chromatographic peaks.

[0046] ​​​​​​(16) The sub-fraction F6-8-11 obtained in step (15) was subjected to fifth dimensional separation and purification by C18HCE (reversed phase column), mobile phase A was (0.1% v / v) formic acid-methanol, B was (0.1% v / v) formic acid-water, gradient elution condition was 0-15 min: 24% A, and compound 11 (tR= 12.8 min) was collected according to the chromatographic peak, named as Corybungine K. R

[0047] (17) The sub-fraction F8 obtained in step (3) was subjected to third dimensional separation and purification by C8CE (reversed phase column), mobile phase A was (0.1% v / v) 25% ammonia water-methanol, B was (0.1% v / v) 25% ammonia water-water, gradient elution condition was 0-5 min: 5% A, 5-50 min: 5-95% A, 50-60 min: 95% A, and 14 sub-fractions F8-1 to F8-14 were collected according to the chromatographic peak.

[0048] (18) The sub-fraction F8-10 obtained in step (17) was subjected to fourth dimensional separation and purification by FC8HL (reversed phase column), mobile phase A was (20 mM) ammonium formate-95% methanol / water (v / v), B was (20 mM) ammonium formate-water, gradient elution condition was 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, 13.88-32 min: 100% A, and 7 sub-fractions F8-10-1 to F8-10-7 were collected according to the chromatographic peak.

[0049] (19) The sub-fraction F8-10-1 obtained in step (18) was subjected to fifth dimensional separation and purification by C18HCE (reversed phase column), mobile phase A was (0.1% v / v) formic acid-acetonitrile, B was (0.1% v / v) formic acid-water, gradient elution condition was 0-15 min: 20% A, and compound 12 (tR= 12.1 min) was collected according to the chromatographic peak, named as Corybungine L. R

[0050] (20) The sub-fraction F8-11 obtained in step (17) was subjected to fourth dimensional separation and purification by FC8HL (reversed phase column), mobile phase A was (20 mM) ammonium formate-95% methanol / water (v / v), B was (20 mM) ammonium formate-water, gradient elution condition was 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, 13.88-32 min: 100% A, and 8 sub-fractions F8-11-1 to F8-11-8 were collected according to the chromatographic peak.

[0051] ​​(21) The subfraction F8-11-2 obtained in step (20) was subjected to fifth dimensional separation and purification by C18HCE (reversed-phase column), mobile phase A was (0.1% v / v) formic acid-acetonitrile, B was (0.1% v / v) formic acid-water, gradient elution condition was 0-10 min: 45% A, and compound 10 (tR= 6.4 min) was collected according to the chromatographic peak, named as Corybungine J. R = 6.4 min), named as Corybungine J.

[0052] (22) The subfraction F8-13 obtained in step (17) was subjected to fourth dimensional separation and purification by FC8HL (reversed-phase column), mobile phase A was (20 mM) ammonium formate-95% methanol / water (v / v), B was (20 mM) ammonium formate-water, gradient elution condition was 0-2.38 min: 50% A, 2.38-13.88 min: 50%-100% A, 13.88-32 min: 100% A, and seven subfractions F8-13-1 to F8-13-7 were collected according to the chromatographic peak.

[0053] (23) The subfraction F8-13-2 obtained in step (22) was subjected to fifth dimensional separation and purification by C18HCE (reversed-phase column), mobile phase A was (0.1% v / v) formic acid-methanol, B was (0.1% v / v) formic acid-water, gradient elution condition was 0-10 min: 30% A, and compound 6 (tR= 4.6 min) was collected according to the chromatographic peak, named as Corybungine F. R = 4.6 min), named as Corybungine F.

[0054] (24) The above compound has the following physicochemical properties and spectroscopic characteristics:

[0055] Corybungine A: brown powder; UV (MeOH) λ max (log ε) 305 (4.11), 283 (4.22), 218 (4.46), 202 (4.50) nm; ECD (MeOH) λ max (Δε) λ 307 (+ 0.83), λ 275 (+ 0.35), λ 239 (- 3.79), λ 217 (+ 1.45); HRESIMS m / z 418.1652 [M+H] + (calcd for C 25 H 24 NO3, 418.1654).

[0056] Corybungine B: brown powder; UV (MeOH) λ max (log ε) 307 (4.05), 281 (4.08), 221 (4.39), 202 (4.40) nm; ECD (MeOH) λ max (Δε) λ 309 (+0.30), λ 279 (+0.20), λ 238 (-1.39), λ 212 (+0.68); HRESIMS m / z 418.1663 [M+H] + (calcd for C 25 H 24 NO3, 418.1654).

[0057] Corybungine C: brown powder; UV (MeOH) λ max (log ε) 325 (3.42), 265 (3.92), 207 (4.29) nm; ECD (MeOH) λ max (Δε) λ 272 (+0.78), λ 232 (-3.64), λ 212 (+0.74); HRESIMS m / z 458.1801 [M+H] + (calcd for C 24 H 28 NO8, 458.1815).

[0058] Corybungine D: black powder; UV (MeOH) λ max (log ε) 317 (4.26), 263 (4.64), 205 (4.80) nm; ECD (MeOH) λ max λ 233 (-0.52); HRESIMS m / z 446.1817 [M+H] + (calcd for C 23 H 28 NO8, 446.1815).

[0059] Corybungine E: black powder; UV (MeOH) λ max (log ε) 308 (3.39), 267 (3.76), 223 (4.28) nm; ECD (MeOH) λ max (Δε) λ 269 (-0.80), λ235 (+2.21); HRESIMS m / z 376.1310 [M+H] + (calcd for C 20 H 22 35 ClNO4, 376.1316); m / z 378.1286 [M+H] + (calcd for C 20 H 22 37 ClNO4, 378.1286).

[0060] Corybungine F: Black powder; UV(MeOH) λ max (logε) 317(3.18), 279(3.58), 216(3.85) nm; ECD(MeOH) λ max (Δε) λ 276 (+0.38), λ 236 (-2.17), λ 212 HRESIMS m / z 328.1543 [M+H][[ID=​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​,462.1911). Corybungine I: brown powder; UV(MeOH)λ max (logε)283(4.14),254(4.52),210(4.61)nm; HRESIMS m / z554.2168[M] + (calcd for C 33 H 32 NO7 + ,554.2173). Corybungine J: brown powder; UV(MeOH)λ max (logε)293(3.58),253(3.82),220(4.23),210(4.22)nm; HRESIMS m / z 402.1549[M+H] + (calcd forC 21 H 24 NO7,402.1553); m / z 424.1368[M+Na] + (calcd for C 21 H 23 NNaO7,424.1372).

[0063] Corybungine K: Yellow powder; UV(MeOH)λ max (logε)312(3.58),257(4.29),210(4.30)nm; HRESIMS m / z366.0970[M] + (calcd for C 20 H 16 NO6 + ,366.0972). Corybungine L: yellow powder; UV(MeOH)λ max (logε)289(3.15),235(3.34),207(3.64)nm; HRESIMS m / z 416.1341[M+H] + (calcd for C 21 H 22 NO8, 416.1345).

[0064] Compounds 1-12 1 H-NMR and 13 The C-NMR data are shown in Tables 1, 2, 3, and 4. 1 H-NMR and 13 The C-NMR spectrum is as follows: Figure 1 As shown, the key two-dimensional NMR information of the compound is as follows:Figure 2 as shown.

[0065] Table I: H-NMR data (δ 1 H-NMR data (δ H , J in Hz, MeOD)

[0066]

[0067]

[0068] Table II: C-NMR data (δ 13 C-NMR data (δ C , MeOD)

[0069] No. 1 2 3 4 5 6 1 144.0,C 144.2,C 145.0,C 147.2,C 144.6,C 143.4,C 1a 117.1,C 117.4,C 117.4,C 128.4,C 126.1,C 121.3,C 1b 123.0,C 123.8,C 124.7,C 129.2,C 128.2,C 122.2,C 2 149.5,C 149.5,C 149.6,C 152.0,C 153.4,C 147.0,C 3 107.8, CH 108.1, CH 108.6, CH 116.5, CH 113.1, CH 113.6, CH 3a 126.1,C 126.4,C 126.0,C 129.4,C 130.3,C 122.2,C 4 27.7, CH2 28.0, CH2 27.7, CH2 30.1, CH2 29.0, CH2 27.1, CH2 5 54.0, CH2 54.1, CH2 54.0, CH2 56.6, CH2 53.6, CH2 54.1, CH2 6a 63.4, CH 63.5, CH 63.1, CH 66.6, CH 63.6, CH 63.7, CH 7 33.1, CH2 32.8, CH2 26.0, CH2 35.5, CH2 32.2, CH2 26.3, CH2 7a 130.8,C 126.4,C 123.9,C 137.1,C 122.5,C 128.3,C 8 114.0, CH 119.6,C 155.8,C 129.4, CH 144.0,C 146.5,C 9 152.3,C 148.1,C 118.0, CH 128.8, CH 150.8,C 152.9,C 10 146.5,C 151.1,C 129.1, CH 128.0, CH 113.7, CH 111.8,C 11 120.3, CH 113.2 123.1, CH 129.2, CH 127.3,C 126.3,C 11a 124.5,C 126.9,C 132.7,C 133.2,C 124.2,C 127.0,C <![CDATA[-OCH2O-]]> 102.6, CH2 102.7, CH2 102.7, CH2 OCH3-1 61.2, CH3 62.4, CH3 OCH3-2 56.5, CH3 OCH3-8 61.2, CH3 OCH3-9 56.5, CH3 56.9, CH3 56.2, CH3 OCH3-10 56.7, CH3 <![CDATA[N-CH3]]> 42.0, CH3 42.3, CH3 42.1, CH3 43.4, CH3 41.8, CH3 1′ 152.1,C 151.2,C 103.7, CH 102.7, CH 2′ 119.3, CH 120.7, CH 74.9, CH 75.0, CH 3′ 116.9, CH 117.1, CH 78.1, CH 78.3, CH 4′ 153.9,C 154.7,C 71.4, CH 71.5, CH 5′ 116.9, CH 117.1, CH 78.3, CH 78.4, CH 6′ 119.3, CH 120.7, CH 62.5, CH2 62.6, CH2

[0070] Table III: H-NMR data (δ 1 H-NMR data (δ H , J in Hz, MeOD)

[0071]

[0072]

[0073] Note: * indicates signal obscured by solvent.

[0074] Table IV: C-NMR data (δ 13 C-NMR data (δ C , MeOD)

[0075]

[0076]

[0077] Note: * indicates signal obscured by background. a indicates signal read from HMBC correlation.

[0078] Activity test examples:

[0079] Cell culture and transfection experiments:

[0080] Chinese hamster ovary (CHO) cells were obtained from the Nation Collection of Authenticated Cell Cultures. CHO cells were cultured in F-12K medium supplemented with 10% fetal bovine serum (FBS) at 37 °C in 5% CO2 in air. The generation of CHO cells stably expressing the dopamine D2 receptor was achieved using standard lipofectamine methods. Human D2 receptor was transfected into CHO cells as follows: CHO cells were transfected with 8 μg of D2 plasmid mixed with 24 μL of lipofectamine 2000 reagent (Invitrogen). Twenty-four hours after transfection, cells were cultured in the presence of 600 μg / mL zeocin for two weeks. Untransfected cells would die during these periods, and then cell cloning was performed using complete medium containing 600 μg / mL zeocin. Stable clonal cells (CHO-D2 cells) were further cultured in modified F-12K medium supplemented with 10% fetal bovine serum (FBS) and 300 μg / mL Zoecin at 37 °C in 5% CO2 in air for two more weeks.

[0081] Dynamic mass redistribution assay (DMR):

[0082] The Epic system (Corning Inc.) is a wavelength interrogation read system tailored for the resonant waveguide grating (RWG) biosensor in microtiter plates, which was used for DMR assay. For whole cell DMR experiments, CHO-D2 cells were seeded directly into the Epic 384-well biosensor plate at 15,000 cells per well and incubated overnight to form a confluent monolayer in cell culture medium. The culture medium in the 384-well biosensor plate was then replaced with 30 μL of Hank's balanced salt solution (1x HBSS), and measurements were taken after a further 1 h incubation inside the system. Compounds were dissolved in dimethyl sulfoxide at a concentration of 100 mM and diluted in HBSS buffer to different concentrations before being automatically added to the wells. Meanwhile, 6.25 nM of dopamine was prepared as a D2 receptor agonist probe.

[0083] For the determination of the dopamine D2 dose activity of compounds 1, 3, 8, a 2 minute baseline was first established, then the first step was to add the compounds in a gradient dilution, the highest final concentration was 100 μM, 4-fold gradient dilution, 7 concentration points (100 μM, 25 μM, 6.25 μM, 1.563 μM, 0.391 μM, 0.098 μM, 0.024 μM), and the DMR response triggered by the compounds within 1 hour was recorded. The second step was to add the drug: reestablish the baseline, then add the D2 agonist dopamine with a final concentration of 6.25 nM per well, and then record the DMR response triggered by dopamine and the test compound within 1 hour, and the results are shown in Table 5. Figure 3 All the IC 50 values described in the present application are calculated based on the maximum DMR signal within 1 hour after the compound stimulation, and the DMR response induced by the assay buffer containing 0.1% dimethyl sulfoxide is used as a negative control, and all the DMR responses are background corrected.

[0084] Table 5 Dopamine D2 receptor antagonistic activity data of compounds

[0085]

[0086] The dopamine D2 receptor antagonistic activity data of Corybungine A, C, H are shown in Table 5, and the test results show that the three compounds all cause dopamine desensitization, indicating that they have obvious D2 receptor antagonistic activity. Therefore, the compounds of the present application can be used to develop lead compounds for the treatment of nervous system diseases such as pain, schizophrenia, Lesch-Nyhan syndrome, etc.

Claims

1. An isoquinoline alkaloid compound, characterized in that: the structural formula is as follows: 。 2. A process for the preparation of a compound according to claim 1, characterized in that: comprising the following steps: (1) medicinal material extraction: taking dry whole grass with roots of Kudou- ding, crushing to obtain medicinal material powder; heating extraction with 50%-90% volume concentration ethanol at 60-70°C for 6-24 hours, filtering the extraction liquid with a 70-80 mesh filter screen, and removing the solvent by rotary evaporation to obtain an extract; dissolving the extract with 0.01-0.2 mol / L sulfuric acid, adjusting the pH to 2-3, adding petroleum ether in a volume ratio of 1:1-1:2 to extract, and taking the water layer; adjusting the pH of the obtained water layer to 9-10 with 0.01-2 mol / L sodium hydroxide, adding dichloromethane in a volume ratio of 1:1-1:2 to extract, and obtaining the crude base; (2) purifying the crude base obtained in step (1) by first-dimensional separation with a C18HCE reverse phase chromatographic column, using 0.1%-2% formic acid-methanol as mobile phase A and 0.1%-2% formic acid-water as mobile phase B, gradient elution conditions being 25% A for 0-8 min, 40% A for 8-23 min, 80% A for 23-38 min, and 100% A for 38-55 min, taking the sample at 4.2-26 min, and concentrating to obtain the total alkaloid sample of Kudou-ding; (3) purifying the total alkaloid of Kudou-ding obtained in step (2) by second- dimensional separation with a C18HCE reverse phase chromatographic column, using 0.1%-2% formic acid-methanol as mobile phase A and 0.1%-2% formic acid-water as mobile phase B, gradient elution conditions being 20% A for 0-5 min, 20-45% A for 5-50 min, and 95% A for 50-60 min, and collecting 11 sub-fractions F1-F11 according to the chromatographic peaks; (4) purifying the sub-fraction F4 obtained in step (3) by third-dimensional separation with a C8CE reverse phase chromatographic column, using 0.1%-2% ammonia water-methanol as mobile phase A, wherein the mass concentration of ammonia water is 20-25%, and using 0.1%-2% ammonia water-water as mobile phase B, wherein the mass concentration of ammonia water is 20-25%, gradient elution conditions being 5% A for 0-5 min, 5-95% A for 5-50 min, and 95% A for 50-60 min, and collecting 15 sub-fractions F4-1-F4-15 according to the chromatographic peaks; (5) purifying the sub-fraction F4-13 obtained in step (4) by fourth-dimensional separation with an FC8HL reverse phase chromatographic column, using 5-50 mM ammonium formate-95% volume concentration methanol / water as mobile phase A and 5-50 mM ammonium formate-water as mobile phase B, gradient elution conditions being 50% A for 0-2.38 min, 50%-100% A for 2.38-13.88 min, and 100% A for 13.88-32 min, and collecting 10 sub-fractions F4-13-1-F4-13-10 according to the chromatographic peaks; (6) The sub-fraction F4-13-4 obtained in step (5) is subjected to fifth dimensional separation and purification by a C18HCE reversed-phase chromatographic column, the mobile phase A is 0.1%-2% formic acid-methanol by volume concentration, B is 0.1%-2% formic acid-water by volume concentration, the gradient elution condition is 0-5 min: 25% A, 5-15 min: 25-45% A, 15-20 min: 45% A, and compound 8 is collected according to the chromatographic peak, t R = 13.40-13.90 min, named as Corybungine H.

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