Preparation and use of nmda receptor inhibitors based on the ginkgolide skeleton

Abstract

The application discloses preparation and application of an NMDA receptor inhibitor based on a gimalin skeleton, relates to the technical field of medicines, and particularly relates to a preparation method of gimalin derivatives shown in general formula I and II and an effect of the derivatives as neuroprotective agents, and mainly applies to anti-cerebral ischemia.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the preparation of a gemmaconone derivative and its application as a GluN2B-NMDA receptor antagonist. Background Technology

[0002] Alzheimer's disease is a heterogeneous disease caused by aging, genetics, and environmental factors, and it is also a common neurodegenerative disease. Current research indicates that the pathogenesis of Alzheimer's disease is related to dysfunction of the neurotransmitter system, particularly the cholinergic system, leading to cognitive impairment and neuropsychiatric disorders in patients. Its common clinical features include progressive memory loss, cognitive impairment, and abnormal behavioral activities.

[0003] Glutamate, as an important excitatory neurotransmitter, has receptors that are important excitatory amino acid receptors in the human body, including two main categories: ionotropic glutamate receptors and metabolic glutamate receptors. Ionotropic glutamate receptors can be further divided into NMDA receptors, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, and erythropoietin receptors, depending on the exogenous agonist.

[0004] As an important glutamate receptor, NMDA receptors regulate neuronal survival and play a crucial role in dendritic and axonal development, synaptic plasticity, neuronal circuit formation, and learning and memory activities. Furthermore, NMDA receptors are involved in multiple important physiological processes of the nervous system and are closely related to various neurodegenerative diseases such as pain, stroke, epilepsy, schizophrenia, depression, and Alzheimer's disease. Overactivation of NMDA receptors can lead to neuronal damage and even apoptosis, causing central nervous system dysfunction and subsequently triggering the aforementioned diseases. Therefore, NMDA receptors are considered to have significant potential for treating neurodegenerative diseases and are important therapeutic targets for various central nervous system disorders.

[0005] NMDA receptors are typically heterotetramers composed of two NR1 subunits and two NR2 subunits. NMDA receptors possess the following characteristics that significantly distinguish them from other glutamate receptors: 1) they are widely distributed both synapticly and extrasynaptactically; 2) they exhibit voltage-dependent Mg2+. 2+ 3) Blocking effect; on Ca 2+ 4) The slow release of glutamate leads to a slower dynamic process involving the NMDA receptor; 5) The activation of the NMDA receptor also requires the participation of glycine or serine in addition to glutamate; 6) The NMDA receptor itself is equipped with a series of regulatory sites, making it abnormally sensitive to the extracellular microenvironment.

[0006] Sesquiterpenoids extracted from plant volatile oils often possess anti-inflammatory, anticancer, antiviral, analgesic, and antitussive effects. Gemarone, an important component extracted from turmeric volatile oil, also exhibits antitumor, antibacterial, and anti-inflammatory pharmacological activities. Various in vitro and in vivo studies have shown that gemmarone is an effective anticancer compound, demonstrating significant pharmacological activity against breast cancer, liver cancer, prostate cancer, glioma cells, and gastric cancer. It can arrest the cell cycle and induce apoptosis in cancer cells by regulating different intracellular signal transduction pathways. Gemarone significantly improves motor dysfunction and spatial learning and memory impairment in mice induced by traumatic brain injury, and dose-relatedly reduces neuronal apoptosis and microglia activation. It also upregulates the expression of the antioxidant protein Nrf2 and inhibits the expression of the inflammatory protein p-p65. Studies have also confirmed that gemimazone can significantly reduce brain water content, infarct volume, and neurological deficits in a rat model of transient middle cerebral artery occlusion / reperfusion injury. Gemazone treatment also decreased MDA content and increased glutathione (GSH), SOD, and GSH-Px activities. Gemazone reduced Caspase-3 and Bax levels while increasing the expression of messenger RNA, Bcl-2, and p-Akt. Therefore, the neuroprotective effect of gemimazone is related to its regulation of neuroinflammation, oxidative stress, and anti-apoptosis.

[0007] NMDA receptors, as potential drug targets and their central nervous system-specific roles, have long attracted researchers' attention. These ligand-gated, voltage-dependent ion channels are widely distributed in the central nervous system. NMDA receptors can translate specific neuronal activity patterns into long-term changes in synaptic structure and function, thereby controlling memory formation and brain plasticity. Gemmazone, as a natural product, exhibits a variety of significant pharmacological effects. However, its simple structure results in limited interactions with target proteins, necessitating further structural optimization to achieve optimal neuroprotective effects. Therefore, research on the natural product gemmazone and its derivatives is of significant scientific importance, providing a foundation for the discovery of new and innovative drugs. Summary of the Invention

[0008] The purpose of this invention is to design a new type of GluN2B-NMDA receptor antagonist that can interact with the NMDA receptor cavity, based on the binding mode of the traditional selective GluN2B-NMDA receptor antagonist, effendil.

[0009] Another object of the present invention is to provide a method for preparing a gemmaconone derivative.

[0010] To achieve the objectives of this invention, the following technical solution is adopted:

[0011] This invention relates to gemcitabine derivatives having the following general formulas I and II, or pharmaceutically acceptable salts thereof, or optically active forms thereof:

[0012]

[0013] Wherein, R1, R2, R3, R4, R5, R6, R7, R8, and R9 are any one of the following groups: (1) H atom, (2) hydroxyl group, (3) nitro group, (4) amino group, (5) halogen group, (6) cyano group, (7) halo-C1-C6 alkyl group, (8) C1-6 alkyl group, C3-8 cycloalkyl group, C2-6 alkenyl group, C2-6 alkynyl group, C1-18 alkoxycarbonyl group, C1-6 alkoxy group, C2-6 alkenyloxy group, C2-6 alkynyloxy group, C3-8 cycloalkenyl group, (9) A carbonyl group substituted in various ways, which is substituted by one or more substituents selected from H atom, hydroxyl group, C1-6 alkyl group, amino group, C1-6 alkylamino group, C1-6 alkoxy group, and C3-8 cycloalkyl group; (10) An amino group substituted in various ways, which is substituted by one or more substituents selected from H atom, C1-6 alkyl group, C2-6 alkenyl group, C2-6 alkynyl group, C2-6 alkenylsulfonyl group, C1-6 alkyl carbonyl group, C2-6 alkenyl carbonyl group, and C2-6 alkynyl carbonyl group. The halogens shown include fluorine, chlorine, and bromine.

[0014] Preferably, R1, R2, R3, R4, R5, R6, R7, R8, and R9 are any one of the following groups: H atom, hydroxyl group, nitro group, amino group, halogen, cyano group, halogenated C1-C6 alkyl group, C1-6 alkyl group, and C3-8 cycloalkyl group.

[0015] More preferably, R1, R2, R3, R4, R5, R6, R7, R8, and R9 are any one of the following groups: H atom, hydroxyl group, halogen, halogenated C1-C6 alkyl group, or C1-6 alkyl group.

[0016] Optically active, the isomers include R and S configurations.

[0017] Most preferably, the present invention provides the following series of gemmaconone derivatives:

[0018]

[0019]

[0020] The pharmaceutically acceptable salt refers to the salt formed by a compound of general formula I or II with an acid selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid, or aspartic acid.

[0021] The synthetic route for preparing the gemmaconone derivative or its optically active form or racemate as shown in formula I or II of the present invention, and the diastereomeric mixture is as follows: wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are the corresponding groups at the corresponding positions of compounds 1a to 3c.

[0022] Route 1: Synthesis of 3a to 3c

[0023] Starting with substituted or unsubstituted benzaldehyde, a nitration reaction is carried out to obtain intermediate a4. Intermediate a4 undergoes a reductive amination reaction with L-glutamic acid to obtain intermediate a7. Intermediate a7 is cyclized in ethanol under reflux to obtain intermediate a10. Intermediate a10 undergoes a secondary cyclization under the catalysis of FeCl3·6H2O to obtain the key intermediate F1. R6, R7, R8, and R9 are the corresponding functional groups at the corresponding positions of compounds 3a–3c.

[0024]

[0025] Gemmaconone A1 is reduced to reduce the ketone carbonyl group to a hydroxyl group to obtain intermediate A2. Intermediate A2 and intermediate F1 undergo ester condensation under the action of a condensing agent to obtain final products 3a to 3c.

[0026]

[0027] Route 2: Synthesis of 1a to 2c

[0028] The substituted benzoic acid and alanine methyl ester salt were amide-condensed in the presence of a condensing agent to obtain intermediate D1. Intermediate D1 was then demethylated in a sodium hydroxide methanol solution to yield carboxylic acid intermediate E1. Intermediate E1 was esterified with the reduction product (A2) of gimazone to give products 1a–2c. Alanine methyl ester salts include D-form and L-form; the L-form was used to prepare compounds 1a–1c, and the D-form was used to prepare compounds 2a–2c. R1, R2, R3, R4, and R5 are the corresponding functional groups at the corresponding positions in compounds 1a–2c.

[0029]

[0030] A pharmaceutical composition comprising one or more of the following: a gemmaconone derivative of formula I or II and its stereoisomers, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

[0031] The dosage form of the pharmaceutical composition is selected from one or more of the following: tablets, capsules, injections, suppositories, patches, inhalable powder formulations, suspensions, emulsions, and ointments.

[0032] The use of gemmaconone derivatives of general formula I or II, or pharmaceutically acceptable salts thereof, or optically active forms thereof, or the pharmaceutical compositions thereof, in the preparation of medicines for the treatment and / or prevention of Alzheimer's disease.

[0033] The use of the gemmaconone derivatives of general formula I or II, or pharmaceutically acceptable salts thereof, or optically active forms thereof, or the pharmaceutical compositions thereof, in the preparation of GluN2B-NMDA receptor antagonists.

[0034] Advantages of this invention:

[0035] This invention synthesizes a series of neo-gemmazone derivatives, through... 1 H-NMR, 13 The structures of the compounds were confirmed by C-NMR and ESI-HRMS, and their cell-protective activities were evaluated. Pharmacological activity tests revealed that all gemcitabine derivatives enhanced cell protection rates, with compound 3a exhibiting the best activity, showing superior activity compared to the positive control drug efendil. Furthermore, the mechanism of action of compound 3a was determined through calcium ion influx experiments. Attached Figure Description

[0036] Figure 1 The protective rates (%) of compounds 1a-3c in the PC12 cell injury model of the present invention are shown in the figures.

[0037] Figure 2 Green fluorescence images of PC12 cells at different time points in this embodiment of the invention show intracellular Ca2+. 2+ Changes in concentration;

[0038] Figure 3 For the embodiment of the present invention Ca 2+ Fluorescence intensity curves for each group during the influx experiment detection period. Detailed Implementation

[0039] The following examples will provide a better understanding of the compounds of the present invention and their preparation. These examples are intended to illustrate, rather than limit, the scope of the invention.

[0040] Example 1: (S)-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline-1-carboxylic acid (F1-1)

[0041] Step A: Preparation of 2-nitrobenzaldehyde (a4)

[0042] Under ice bath conditions, potassium nitrate (3.90 g, 0.039 mol) was added to concentrated sulfuric acid (30 mL, d = 1.83 g / mL), and the reaction temperature was maintained at 0-3℃. Benzaldehyde (6.93 g, 0.037 mol) was slowly added dropwise, and the reaction was carried out at below 5℃ for 2 hours. After the reaction was completed, the reaction solution was poured into 160 mL of an ice-water mixture, and a white solid precipitated. The solid was filtered, dried, and separated by column chromatography (PE:EA = 30:1) to obtain 4.89 g of white needle-like crystals, with a yield of 74.63%.

[0043] Step B: Preparation of (S)-2-(2-nitrobenzylamino)-glutaric acid (a7)

[0044] Dissolve 2.38 g (0.06 mol) of NaOH in 20 mL of water under ice bath conditions. Then, slowly add L-glutamic acid (4.41 g, 0.03 mol) in portions to the NaOH solution under ice bath conditions. Separately, dissolve 3.71 g (0.02 mol) of o-nitrobenzaldehyde in 30 mL of anhydrous ethanol under stirring. After complete dissolution, slowly add this solution dropwise to the L-glutamic acid reaction solution under ice bath conditions, controlling the addition time to approximately 1 hour. After the addition is complete, transfer the reaction solution to room temperature and continue stirring for 1 hour. Then, return the solution to the ice bath to cool, and slowly add 0.76 g (0.02 mol) of NaBH4 in portions to the reaction solution. After the addition is complete, return the solution to room temperature and continue stirring for approximately 3 hours until the reaction solution becomes clear. After the reaction was monitored by TLC, a small amount of insoluble impurities in the reaction solution were removed by filtration using diatomaceous earth as the medium. The filtrate was concentrated to approximately 20 mL, extracted with ethyl acetate, and the organic layer was discarded. The pH of the aqueous layer was adjusted to 4-5 with 1 mol / L hydrochloric acid, resulting in the precipitation of a large amount of white solid. The mixture was stirred for another 30 min at room temperature, filtered, and the filter cake was washed twice with 30 mL of water. After drying, a white, lustrous powdery solid was obtained, weighing 4.39 g, with a yield of 78.36%.

[0045] Step C: Preparation of (S)-1-(2-nitrobenzylamino)-5-oxopyrrolidine-2-carboxylic acid (a10).

[0046] (S)-2-(2-nitrobenzylamino)glutaric acid (4.39 g, 0.016 mol) was added to anhydrous ethanol (45 mL), and the mixture was heated under reflux for 2 h. The reaction solution became almost clear. The solution was filtered while hot to remove a small amount of insoluble matter. After cooling, a white solid precipitated. The solid was filtered, the filter cake was collected, and the filtrate was concentrated to about 15 mL. The two filter cakes were then combined to give 4.01 g of white solid, with a yield of 97.23%.

[0047] Step D: Preparation of (S)-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline-1-carboxylic acid (F1)

[0048] (S)-1-(2-nitrobenzylamino)-5-oxopyrrolidine-2-carboxylic acid (4.01 g, 0.015 mol) was added to anhydrous ethanol (55 mL), heated to dissolve, and then cooled slightly. Activated carbon (0.530 g) and FeCl3·6H2O (0.20 g, 0.75 mmol) were added sequentially. The mixture was then refluxed vigorously for 72 h after reaching 70 °C. The mixture was filtered, and the filtrate was concentrated, leaving approximately 15 mL. 20 mL of acetone was slowly added dropwise with stirring. The mixture was then filtered under vacuum and dried to give 1.86 g of a white solid, with a yield of 56.78%.

[0049] Example 2: (S)-7-chloro-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline-1-carboxylic acid (F1-2)

[0050] The preparation method is the same as above.

[0051] Example 3: (S)-7-bromo-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline-1-carboxylic acid (F1-3)

[0052] The preparation method is the same as above.

[0053] Example 4: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2S)-2-(4-hydroxybenzamide)propionate (1a)

[0054] Step A: Preparation of (S)-2-(4-hydroxybenzamide)propionate methyl ester (D1)

[0055] 1.93 g (0.014 mol) of 4-hydroxybenzoic acid was dissolved in 10 mL of ultra-dry DMF. Under an ice-water bath, 8.38 g (0.022 mol) of HATU was slowly added, followed by dropwise addition of DIPEA (6.07 mL (0.037 mol)). After the addition was complete, the mixture was stirred under an ice-water bath for 15 min, then moved to room temperature for 30 min. The reaction was monitored by TLC, and the reaction solution was retained. Subsequently, 2.09 g (0.015 mol) of L-alanine methyl ester hydrochloride was added under an ice-water bath. The mixture was stirred for 15 min, then moved to room temperature for 2 h, and the reaction was monitored by TLC. The reaction solution was then slowly poured into 100 mL of water and extracted with DCM. The organic layers were combined and extracted with saturated brine. The organic layer was retained, dried, concentrated, and then subjected to column chromatography (DCM:MeOH = 50:1) to give 2.90 g of a white solid, with a yield of 89.05%.

[0056] Step B: Preparation of (S)-2-(4-hydroxybenzamide)propionic acid (E1)

[0057] At room temperature, the intermediate D1 (2.50 g, 0.011 mol) obtained above was dissolved in a methanol solution of 4 mol / L NaOH. After stirring for 1 h, the reaction was monitored by TLC. The reaction solution was then concentrated to dryness, and the pH was adjusted to 3-4 with 15% dilute hydrochloric acid. The solution was then extracted with ethyl acetate, and the organic layers were combined. The organic layer was extracted with saturated brine, and the organic layer was retained. After drying and concentration, a white solid was obtained, weighing 2.12 g, with a yield of 90.32%.

[0058] Step C: Preparation of (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecane 3,7-dien-1-ol (A2)

[0059] LiAlH4 (0.38 g, 0.010 mol) was placed in a 50 mL round-bottom flask, and 10 mL of tetrahydrofuran (THF) was added to form a suspension. Separately, gimafenone (2.00 g, 0.0092 mol) was dissolved in 5 mL of THF. Under ice bath conditions, the gimafenone THF solution was slowly added dropwise to the LiAlH4 THF suspension, controlling the addition time to approximately 2 min. After the addition was complete, the mixture was moved to room temperature and stirred for 10 min, with TLC monitoring of the reaction. Subsequently, 1 mL of water, 2 mL of 15% NaOH aqueous solution, and 3 mL of water were added sequentially to the reaction solution to quench the reaction. The mixture was extracted with petroleum ether, retaining the organic layer. After drying with anhydrous sodium sulfate, the residue was filtered and evaporated to dryness to obtain 1.85 g of a milky white oily substance, with a yield of 91.72%.

[0060] Step D: Preparation of (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2S)-2-(4-hydroxybenzamide)propionate (1a)

[0061] Intermediate E1 (200 mg, 0.97 mmol) was dissolved in 10 mL of ultra-dry DCM solution. EDCI (277 mg, 14.62 mmol) and DMAP (295 mg, 0.78 mmol) were slowly added under an ice-water bath. The mixture was then stirred for 15 min under an ice-water bath. Next, intermediate A2 (214 mg, 0.97 mmol) was added under an ice-water bath, and the mixture was stirred for another 15 min. The reaction was then allowed to continue at room temperature for 2 h, and the reaction was monitored by TLC. The mixture was then extracted with saturated brine, retaining the organic layer. After drying and concentration, column chromatography (DCM:MeOH = 50:1) yielded 198 mg of a white powder, with a yield of 49.87%.

[0062] 1H NMR(600MHz,Chloroform-d)δ9.68(s,1H),8.70(d,J=7.2Hz,1H),7.97(dd,J=8.6,5.6Hz ,2H),7.31(t,J=8.7Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3 .78(d,J=12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=50.1Hz,2H),2.02(d,J= 6.5Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H),1.40(d,J=7.3Hz,3H). 13 C NMR(150MHz,DMSO-d6)δ170.31,163.57,162.13,160.53,142.83,135.86,131.05,128.16,127.19,1 26.25,125.25,122.34,115.40(d,J=21.1Hz),64.54,44.90,34.49,30.07,24.34.ESI-HRMS:calced for C 25 H 33 NO4, [M+H] + ,found 412.1845.

[0063] Example 5: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2S)-2-(4-fluorobenzamide)propionate (1b)

[0064] In Example 4, step A, 4-hydroxybenzoic acid was replaced with 4-chlorobenzoic acid, and the preparation method was the same as in Example 4, with a yield of 45.82%.

[0065] 1H NMR(600MHz,Chloroform-d)δ8.70(d,J=7.2Hz,1H),7.97(dd,J=8.6,5.6Hz,2H),7.31 (t,J=8.7Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3.78(d,J =12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=50.1Hz,2H),2.02(d,J=65.0 Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=94.5Hz,6H),1.40(d,J=7.3Hz,3H). 13 C NMR (150MHz, DMSO-d6) δ170.20,163.68,143.21,131.54,131.09,130.21,129.09,125.65,122.82,64.61,44.74,30.12,24.25.ESI-HRMS:calced for C 25 H 32 ClNO3,[M+H] + ,found 430.0929.

[0066] Example 6: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2S)-2-(4-chlorobenzamide)propionate (1c)

[0067] In Example 4, step A, 4-hydroxybenzoic acid was replaced with 4-fluorobenzoic acid, and the preparation method was the same as in Example 4, with a yield of 40.58%.

[0068] 1 H NMR(600MHz,Chloroform-d)δ8.70(d,J=7.2Hz,1H),7.97(dd,J=8.7,5.7Hz,2H),7.3 1(t,J=8.8Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3.78(d ,J=12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=5.1Hz,2H),2.02(d,J=6.0 Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H),1.40(d,J=7.3Hz,3H).13 C NMR(150MHz,DMSO-d6)δ170.08,163.64,158.15,131.54,130.21,128.24,127.40,126.3 2,124.86,122.18,114.15,64.71,55.42,44.87,34.50,30.07,24.27.ESI-HRMS:calced for C 25 H 32 FNO3, [M+H] + ,found 414.0805.

[0069] Example 7: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2R)-2-(4-hydroxybenzamide)propionate (2a)

[0070] The synthesis method of compound 2a is the same as that of compound 1a, except that the intermediate L-alanine methyl ester hydrochloride is replaced with the intermediate D-alanine methyl ester hydrochloride, yielding a white powdery solid with a yield of 36.29%.

[0071] 1 H NMR(600MHz,Chloroform-d)δ9.68(s,1H),8.70(d,J=7.2Hz,1H),7.97(dd,J=8.6,5.6Hz ,2H),7.31(t,J=8.7Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3 .78(d,J=12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=5.1Hz,2H),2.02(d,J= 6.0Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H),1.40(d,J=7.3Hz,3H). 13 CNMR(150MHz,DMSO-d6)δ172.47,167.81,134.51,133.85,129.75,129.47,128.79,126.51,12 3.20,116.15,115.22,71.68,49.56,43.84,39.27,25.86,25.65,20.99,20.27,17.23,16.42. ESI-HRMS:calced for C 25 H 33 NO4, [M+Na] +,found434.0471.

[0072] Example 8: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2R)-2-(4-chlorobenzamide)propionate (2b)

[0073] The synthesis method of compound 2b is the same as that of compound 1b, except that the intermediate L-alanine methyl ester hydrochloride is replaced with the intermediate D-alanine methyl ester hydrochloride, yielding a white powdery solid with a yield of 48.29%.

[0074] 1 H NMR(600MHz,Chloroform-d)δ8.70(d,J=7.2Hz,1H),7.97(dd,J=8.6,5.6Hz,2H),7.3 1(t,J=8.7Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3.78(d ,J=12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=5.1Hz,2H),2.02(d,J=6.5 Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H),1.40(d,J=7.3Hz,3H). 13 C NMR (150MHz, DMSO-d6) δ170.20,163.68,143.21,131.54,131.09,130.21,129.09,125.65,122.82,64.61,44.74,30.12,24.25.ESI-HRMS:calced for C 25 H 32 ClNO3,[M+H] + ,found 431.0471.

[0075] Example 9: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecyl 3,7-dien-1-yl-(2R)-2-(4-fluorobenzamide)propionate (2c)

[0076] The synthesis method of compound 2c is the same as that of compound 1c, except that the intermediate L-alanine methyl ester hydrochloride is replaced with the intermediate D-alanine methyl ester hydrochloride, yielding a white powdery solid with a yield of 47.54%.

[0077] 1H NMR(600MHz,Chloroform-d)δ8.70(d,J=7.2Hz,1H),7.97(dd,J=8.7,5.7Hz,2H),7.3 1(t,J=8.8Hz,2H),4.79(s,1H),4.52(d,J=12.8Hz,1H),4.42(t,J=7.3Hz,1H),3.78(d ,J=12.5Hz,1H),3.22–2.47(m,2H),2.40(s,1H),2.23(d,J=5.1Hz,2H),2.02(d,J=6.5 Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H),1.40(d,J=7.3Hz,3H). 13 C NMR(150MHz,DMSO-d6)δ170.08,163.64,158.15,131.54,130.21,128.24,127.40,126.3 2,124.86,122.18,114.15,64.71,55.42,44.87,34.50,30.07,24.27.ESI-HRMS:calced for C 25 H 32 FNO3, [M+H] + ,found 414.1013.

[0078] Example 10: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecane 3,7-dien-1-yl-(S)-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazolin-1-carboxylate (3a)

[0079] Intermediate F1-1 (0.22 g, 1.09 mmol) was dissolved in 10 mL of 99.9% extraturkey in a 25 mL round-bottom flask. The reaction flask was then placed in an ice-water bath. Condensing agents EDCI (0.21 g, 1.09 mmol) and DMAP (0.089 g, 0.73 mmol) were weighed and slowly added to the reaction flask with stirring. After stirring for 20 min in the ice-water bath, intermediate A2 (0.20 g, 0.91 mmol) was added. The mixture was then moved to room temperature and stirred for 2 h. The reaction was monitored by TLC. The reaction solution was then extracted with saturated brine, retaining the organic layer. After drying with anhydrous calcium chloride, the solution was filtered, concentrated, and purified by column chromatography (PE:EA = 10:1) to obtain white needle-like crystals in 31.34% yield.

[0080] 1H NMR(600MHz,Chloroform-d)δ7.30(d,J=7.8Hz,2H),7.17(s,1H),6.99(d,J=7.8Hz,1H),4.86(d,J=8.2Hz,1 H),4.71(d,J=14.3Hz,1H),4.52(d,J=12.8Hz,1H),3.78(d,J=12.5Hz,1H),3.42(s,1H),3.16–2.81(m,1H), 2.58(d,J=6.1Hz,1H),2.46(d,J=5.0Hz,1H),2.40(s,2H),2.36(d,J=8.6Hz,1H),2.23(d,J=5.1Hz,1H),2.0 8(s,2H),1.97(s,2H),1.87(s,1H),1.78(d,J=3.3Hz,1H),1.69(d,J=9.1Hz,3H),1.59(s,3H),1.43(s,4H). 13 C NMR(150MHz,DMSO-d6)δ170.31,163.89,143.36,139.74,131.07,129.22,129.1 3,128.76,126.57,125.72,64.56,44.79,35.40,30.11,24.34.ESI-HRMS:calced forC 27 H 34 N₂O₂, [M+H] + ,419.0701,found 419.0749.

[0081] Example 11: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecane 3,7-dien-1-yl-(S)-7-chloro-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazolin-1-carboxylate (3b)

[0082] The synthesis method of compound 3b is the same as that of compound 3a, except that intermediate F1-1 is replaced with intermediate F1-2. The solid was purified by column chromatography (DCM:MeOH = 20:1) to obtain a brownish-yellow powder with a yield of 35.62%.

[0083] 1H NMR(600MHz,Chloroform-d)δ7.33–7.24(m,2H),6.95(d,J=8.3Hz,1H),4.79(s,1H),4.65(d,J=14.4Hz,1H) ,4.52(d,J=12.8Hz,1H),4.20(d,J=14.4Hz,1H),4.11(t,J=7.2Hz,1H),3.78(d,J=12.5Hz,1H),3.22–2.47(m ,2H),2.40(s,1H),2.56(ddd,J=11.4,6.3,3.3Hz,1H),2.49–2.41(m,1H),2.40(s,1H),2.36(dt,J=17.5,9.0 Hz,1H),2.23(d,J=5.1Hz,2H),2.02(d,J=6.5Hz,2H),1.87(s,1H),1.90–1.65(m,6H),1.51(d,J=9.5Hz,6H). 13 C NMR (150MHz, DMSO-d6) δ169.48,163.96,139.69,129.22,128.41,126.74,123.07,121.42,65.32,44.90,35.36,29.89,24.41.ESI-HRMS:calced for C 27 H 33 ClN2O2,[M+Na] + ,476.0925,found 476.0991.

[0084] Example 12: (3E,7E)-3,7-dimethyl-10-isopropylidene-cyclodecane-3,7-dien-1-yl-(S)-7-bromo-1,2,3,9-tetrahydropyrrolo[2,1-b]quinazolin-1-carboxylate (3c)

[0085] The synthesis method of compound 3c is the same as that of compound 3a, except that intermediate F1-1 is replaced with intermediate F1-3. The brown powder solid was purified by column chromatography (DCM:MeOH = 20:1) with a yield of 28.54%.

[0086] 1H NMR (600MHz, Chloroform-d) δ7.32–7.24(m,2H),6.95(d,J=8.4Hz,1H),4.86(d,J=8.2Hz,1H),4.65(d,J=14.4Hz,1H),4.52(d ,J=12.8Hz,1H),4.20(d,J=14.4Hz,1H),4.11(dd,J=7.9,6.4Hz,1H),3.78(d,J=12.5Hz,1H),3.16–2.81(m,1H),3.16–2.81(m ,1H),2.56(ddd,J=10.2,5.9,2.2Hz,1H),2.44(dd,J=10.2,4.5Hz,1H),2.40–2.31(m,3H),2.23(d,J=5.1Hz,1H),2.04(dtd,J =13.0,8.8,8.3,4.4Hz,1H),1.97(s,2H),1.87(s,1H),1.78(d,J=3.3Hz,1H),1.69(d,J=9.1Hz,3H),1.59(s,3H),1.43(s,4H). 13 C NMR (150MHz, DMSO-d6) δ170.30,163.68,143.25,135.87,131.09,129.11,125. 68,122.82,115.47,64.57,44.76,40.41,34.49,30.10,24.32.ESI-MS:calced for C 27 H 33 BrN2O2,[M+H] + ,498.3,found 498.3.

[0087] Example 13: Pharmacological study of the compounds of the present invention

[0088] (1) In vitro activity test of the compound

[0089] PC12 cells in logarithmic growth phase were seeded at a density of 5000 cells / well and a volume of 100 μL / well in 96-well plates and cultured for 24 h. The test groups were treated with compounds at final concentrations of 0.05, 0.5, and 5 μM / L, while the blank control and NMDA damage model groups were treated with an equal volume of blank culture medium. After 6 h of incubation, the culture medium was discarded and replaced with HEPES buffer (1×HMF, MgSO4). 2+-free, CC0074, Leagene, China). The test and model groups were added to HEPES buffer with a final concentration of 2 mM NMDA, while the blank group was added to normal HEPES buffer. Incubation was carried out for 30 min. The buffer was discarded, and the cells were cultured in normal medium for another 12 h. 10 μL of MTT solution (5 mg / mL) was added to the cell culture medium, and the cells were incubated at 37°C for 4 h. The supernatant was carefully aspirated and discarded. 150 μL / well DMSO was added, and the cells were vortexed to dissolve. The OD value was measured at 490 nm using a microplate reader (ELX 800, Bio-TEK instruments, Inc.), and the cell protection rate was calculated. Efenidyl was used as a positive control.

[0090] Cell protection rate: (1 - (OD test - OD model) / (OD blank - OD model)) × 100%

[0091] Test results are as follows Figure 1 As shown, gimazone exhibits certain neuroprotective activity at low concentrations, but shows significant cytotoxicity at high concentrations. When the gimazone hydroxyl group is incorporated into other structural formulas, the neuroprotective activity of the compounds is significantly better than that of the control drug efendil (3a-3c). The activity of the compounds is not significantly different when H, Br, or Cl atoms are introduced onto the benzene ring in the 1,2,3,9-tetrahydropyrrolo[2,1-b]quinazoline structure. Among them, when there are no substituents on the benzene ring, compound 3a has the best neuroprotective activity, reaching 29.63% at low concentrations. When the substituent of the gimazone hydroxyl group is an alanine-linked benzoic acid, the neuroprotective activity is increased compared to gimazone (1a-2c). The effect of different R / S configurations on the compound activity shows that the configuration of the alanine linker has little effect on the neuroprotective activity of the compound.

[0092] (2) Ca of compound 3a 2+ Inflow Experiment

[0093] PC12 cells in logarithmic growth phase (density 6.0 × 10⁻⁶) were harvested. 4 After incubating (samples / plate) for 24 h in a 3.5 cm confocal plate, treat with or without 0.05 μM of the target compound (efendil or compound 3a obtained above) for 6 h. Add serum-free DMEM fresh medium containing Fluo-4AM (2.5 μM, S1060, Beyotime, China) and incubate for another 0.5 h. Discard the medium and rinse with HEPES buffer (1×HMF, Mg... 2+Cells were washed twice with HEPES buffer (free, CC0074, Leagene, China). After adding 1 ml of HEPES buffer, cells were scanned using a laser scanning confocal microscope (LSMT-PMT, Observer). The scan was performed for 50 seconds (Z1, Zeiss) until the baseline stabilized. Then, NMDA (final concentration 500 μM) or an equal volume of HEPES buffer was added, and the fluorescence intensity was detected after 10 minutes of scanning.

[0094] Test results are as follows Figure 2 As shown, the cytoplasmic Ca of the control group 2+ The concentration remained stable within the detection time, while the cytoplasmic Ca in the model group after the addition of NMDA was observed. 2+ The concentration of intracellular Ca increased significantly after pretreatment with efenidil and 3a. 2+ Inland inflows have decreased significantly. Furthermore, from... Figure 3 The curves show that compound 3a exhibits an inhibitory effect similar to that of levonorgestrel. Compound 3a inhibits Ca2+. 2+ The overall effect of infusion is better than that of fenfenadil.

Claims

1. Gemmaconone derivatives of general formulas I and II, or pharmaceutically acceptable salts thereof, or optically active forms thereof: ； in, R1 and R6 are any one of the following groups: H atom, hydroxyl group, halogen; R2, R3, R4, R5, R7, R8, and R9 are H atoms.

2. The gemimazone derivative or its pharmaceutically acceptable salt or its optically active form according to claim 1, characterized in that, The pharmaceutically acceptable salt refers to the salt formed by the compound represented by general formula I or II and an acid selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid, or aspartic acid; optically active, and isomers include both R and S configurations.

3. The gemimazone derivative according to claim 1, or a pharmaceutically acceptable salt thereof, or an optically active form thereof, characterized in that, Selected from the following compounds: 。 4. The method for preparing the gemmaconone derivative of claim 3 or its pharmaceutically acceptable salt or its optically active form, characterized in that, Includes the following steps: Route 1: Synthesis of 3a~3c; Using substituted or unsubstituted benzaldehyde as a starting material, a nitration reaction is carried out to obtain intermediate a4. Intermediate a4 and... L - Glutamic acid undergoes a reductive amination reaction to obtain intermediate a7. Intermediate a7 is cyclized in ethanol under reflux to obtain intermediate a10. Intermediate a10 undergoes a secondary cyclization under the catalysis of FeCl3·6H2O to obtain the key intermediate F1. ； Gemmaconone A1 is reduced to reduce the ketone carbonyl group to a hydroxyl group to obtain intermediate A2. Intermediate A2 and intermediate F1 undergo ester condensation in the presence of a condensing agent to obtain final products 3a~3c. Among them, R6, R7, R8, and R9 are the corresponding groups at the corresponding positions of compounds 3a~3c. ； Route 2: Synthesis of 1a~2c; The substituted benzoic acid and alanine methyl ester salt were amide condensed in the presence of a condensing agent to obtain intermediate D1. Intermediate D1 was demethylated in the presence of sodium hydroxide methanol solution to obtain carboxylic acid intermediate E1. Intermediate E1 was esterified with the reduction product of gimazone to obtain products 1a-2c. Alanine methyl ester salt includes D-type and L-type. The L-type was used to prepare compounds 1a-1c, and the D-type was used to prepare compounds 2a-2c. R1, R2, R3, R4, and R5 are the corresponding groups at the corresponding positions in compounds 1a-2c. 。 5. A pharmaceutical composition, characterized in that, It comprises one or more of the gemmaconone derivatives of general formula I or II as claimed in claim 1, their stereoisomers or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, characterized in that, The dosage form is selected from tablets, capsules, injections, suppositories, patches, inhalable powders, suspensions, emulsions, and ointments.

7. The use of the gemmaconone derivative of claim 1 or a pharmaceutically acceptable salt thereof or an optically active form thereof, or the pharmaceutical composition of claim 5, in the preparation of a medicament for the treatment and / or prevention of Alzheimer's disease.

8. The use of the gemimadone derivative of claim 1 or a pharmaceutically acceptable salt thereof or an optically active form thereof, or the pharmaceutical composition of claim 5, in the preparation of a GluN2B-NMDA receptor antagonist.

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