Succinimide derivative as well as preparation method and application thereof

Abstract

The invention discloses a succinimide derivative in the field of organic chemistry, and the succinimide derivative has a structural general formula shown as a formula V. During preparation of # imgabs0, a compound shown as a general formula I reacts with a compound shown as a general formula II to obtain a compound shown as a general formula III; and reacting the compound shown as the general formula III with a compound shown as a general formula IV in the presence of a condensing agent to obtain the succinimide derivative V. The succinimide derivative disclosed by the invention can be applied to preparation of medicines for calming, soothing the nerves, promoting intelligence, resisting convulsion, resisting epilepsy, resisting neuropathic pain, resisting anxiety, resisting depression, resisting dementia and / or protecting brain injury.

Description

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on April 19, 2024, with application number 2024104773680 and application name “Succinimide derivatives, preparation methods and applications thereof”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The invention belongs to the field of organic chemistry, and particularly relates to succinimide derivatives and preparation methods and applications thereof. Background Art

[0003] Amber (Ambrum) is the resin of the ancient pine genus of the pine family. It is a representative traditional Chinese medicine for calming the nerves. The Chinese Materia Medica summarizes the effects of amber as "calming the nerves and calming the nerves"; "dispersing blood stasis and stopping bleeding"; "promoting diuresis and relieving stranguria"; and "removing cataracts and improving eyesight". The main ingredients of the prescription "Pediatric Amber Tranquilizing Pills" include amber, cinnabar, musk, gastrodia, plum tablets, and arisaema, etc. This prescription can treat acute and chronic convulsions, fever, convulsions, and coma in children. Succinic acid is a compound with a succinic acid structure, and its structure is widely present in natural products. Modern pharmacological studies have shown that succinic acid derivatives have a wide range of pharmacological activities such as anti-epileptic, anti-inflammatory, anti-tumor and antibacterial.

[0004] Gastrodin (GAS) is a small molecule active compound extracted from the rhizome of Gastrodia elata Blume. Its chemical name is 4-hydroxybenzyl alcohol-4-O-β-D-pyranoglucoside. It is the main pharmacologically active ingredient of the traditional Chinese medicine Gastrodia elata. Studies have shown that Gastrodin has pharmacological effects such as neuroprotection, anti-inflammatory, analgesia, sedation and hypnosis, anticonvulsant, and improvement of behavioral disorders.

[0005] Borneolum Syntheticum has a long history of use in my country. The New Compendium of Materia Medica records that it can treat evil spirits in the heart and abdomen, rheumatism, deafness, improve eyesight, and remove red and cataracts in the eyes. Modern pharmacology shows that borneol has the effects of awakening the mind, clearing away heat and detoxifying, and relieving pain. Tambe et al. found that borneol has the effects of protecting the heart and brain and anticonvulsant, and can significantly inhibit the occurrence of epilepsy in mice ignited by pentylenetetrazol, and inhibit the expression of brain neurogenic stress and neuroinflammatory markers [Tambe R, Naunyn Schmiedebergs Arch Pharmaco, 2016, 389(5):467].

[0006] Based on the excellent pharmacological activities of effective ingredients such as succinic acid, gastrodin, and borneol in sedation, tranquilization, anticonvulsant, and nerve cell protection, a series of succinimide derivatives including succinimide-amino acid-gastrodin ester, succinimide-amino acid-vanillyl alcohol ester, and succinimide-amino acid-borneol ester were synthesized, so as to screen out new chemical drugs with therapeutic effects in anti-epileptic, anticonvulsant, memory improvement, anti-anxiety, anti-depression, anti-dementia and brain damage improvement, which has positive significance. Summary of the invention

[0007] The present invention aims to provide a succinimide derivative having anti-epileptic, anticonvulsant, anti-neuropathic pain, memory improvement, antianxiety, antidepression, anti-dementia and / or brain damage improvement effects.

[0008] The succinimide derivatives in this embodiment have a general structural formula as shown in Formula V:

[0009]

[0010] Where: R 1 H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, branched or straight chain C1-8 alkyl, cycloalkyl C m H 2m-1 (m=3-6), methylcycloalkyl C m+1 H 2m+1 (m=3-6), substituted allyl, substituted phenyl, substituted benzyl, heterocyclic group, methoxymethyl, hydroxymethyl, 2-hydroxyethyl, methylthioethyl, heterocyclic ring; R 2 Methyl, ethyl, isopropyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, branched or straight chain C1-8 alkyl, cycloalkyl C m H 2m-1 (m=3-6), methylcycloalkyl C m+1 H 2m+1 CH 2 (m=3-6), substituted allyl, substituted cinnamyl, rac-2-bornyl, (-)-2-bornyl, (+)-2-bornyl, rac-menthyl, (-)-menthyl, (+)-menthyl, substituted phenyl, substituted benzyl, heterocyclic group, 2-methylpiperidine, dimethylamino, diethylamino, diisopropylamino, 2,2-dimethyl-3-(propylamino)propyl-1-yl, N,2-dimethyl-1-phenylpropyl-2-amino; R 3 is H, methyl, ethyl, phenyl; R 4 is H, methyl, ethyl, phenyl, spirocyclopentyl; or R 3 R 4It constitutes cyclohexyl, phenyl, (1R,4S)-bicyclo[2,2,1]hexane-2-enyl; X is O, NH or piperazinyl; n is 0-3.

[0011] Further, R1 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, 2-methyl-n-butyl, neopentyl, n-hexyl, 2-methyl-n-pentyl, 3-methylpentyl, 4-methylpentyl, 2-hexyl, 4-methylpentane-2-yl, 3-methylpentane-2-yl, 3-pentyl, n-heptyl, n-octyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl, cyclohexanemethyl, allyl, 3-methyl-2-buten-1-yl, substituted phenyl, substituted benzyl, methoxymethyl, hydroxymethyl, 2-hydroxyethyl, methylthioethyl, or aromatic heterocycle.

[0012] Furthermore, the R 1 The structural formula is as follows:

[0013]

[0014] Furthermore, the R 2 The structural formula is as follows:

[0015]

[0016]

[0017] Furthermore, the R 3 or R 4 The structural formula is shown below;

[0018]

[0019] or R 3 R 4 Together they form a cyclohexyl group, a phenyl group or a (1R,4S)-bicyclo[2,2,1]hexane-2-enyl group, the structural formula of which is shown below:

[0020]

[0021] With respect to the above-mentioned succinimide derivatives, the present application also provides a method for preparing succinimide derivatives, comprising the following steps:

[0022] The compound of general formula I reacts with the compound of general formula II to obtain the compound of general formula III; the compound of general formula III reacts with the compound of general formula IV in the presence of a condensing agent to obtain a succinimide derivative V.

[0023]

[0024] Furthermore, the specific preparation process of the compound of formula III is as follows: the compound of formula I and the compound of formula II are added to a reaction container in a molar ratio of 1.0 to 2.0:1.0, and reacted at 70° C. to 100° C. until the compound of formula I and / or the compound of formula II disappear; vacuum concentration is performed to remove the solvent, the residue is precipitated by adding water, filtered, and dried at 30-60° C. to obtain the compound of formula III.

[0025] Furthermore, when preparing succinimide derivatives V: the compound of formula III and the compound of formula IV, a condensing agent, and a catalyst are added to a reaction container in sequence, the molar ratio of the compound of formula III to the compound of formula IV is 1.0:1.0-1.5, and the reaction is carried out at 5°C-40°C until the compound of formula III and / or the compound of formula IV are consumed, pure water is added for washing, ethyl acetate is extracted, and the ethyl acetate is removed by vacuum concentration. The organic phase is dried, vacuum concentrated, and purified by column chromatography.

[0026] Furthermore, the condensing agent is one of DCC, DIC, EDC, BOP, PyBOP, HATU, HBTU, TBTU, ethyl chloroformate, phenyl chloroformate, isopropyl chloroformate, 1-n-propyl phosphoric anhydride, etc., preferably EDC.

[0027] Furthermore, the catalyst is one of Et3N, DIEA, DMAP, DPPY, 2,6-lutidine, DABCO, DBU, etc., preferably DMAP.

[0028] Furthermore, the stereoisomerism of the succinimide derivative or a mixture of different stereoisomers thereof.

[0029] Furthermore, the succinimide derivatives are used in the preparation of sedative, tranquilizing, intelligence-enhancing, anticonvulsant, anti-epileptic, anti-neuropathic pain, antianxiety, anti-depression, anti-dementia and / or brain damage protective drugs.

[0030] Furthermore, the stereoisomers of the succinimide derivatives or mixtures of different stereoisomer compounds thereof are used in the preparation of sedative, tranquilizing, intelligence-enhancing, anticonvulsant, anti-epileptic, anti-neuropathic pain, antianxiety, anti-depression, anti-dementia and / or brain damage protective drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Graph showing the effects of the compound on normal zebrafish.

[0032] Figure 2 Schematic diagram of the effects of compounds on PTZ-induced epileptic seizures in zebrafish.

[0033] Figure 3 This is a comparison of neuronal apoptosis in the CA1 region of mouse hippocampal tissue. DETAILED DESCRIPTION

[0034] The following is further described in detail through specific implementation methods:

[0035] The preparation method of succinimide derivative V, the reaction formula is as follows:

[0036]

[0037] Specific method: Add the compound of formula I and the compound of formula II in a molar ratio of 1.0-2.0:1.0 to a reaction vessel, react at 70-100°C for 3-10 hours, and monitor the reaction on a TLC plate until the raw material disappears. Concentrate under vacuum to remove the solvent, and add water to the residue to precipitate. Filter and dry at 40-60°C to obtain the compound of formula III;

[0038] The compound of formula III and the compound of formula IV, a condensing agent and a catalyst are sequentially added into a reaction vessel in a molar ratio of 1.0:(1.0-1.5):(1.0-1.5):(0.1-0.5), and reacted at 5-40°C for 12-24 hours until the raw materials disappear. Then pure water is added for washing, ethyl acetate is extracted, and the solvent is removed by vacuum concentration. The organic phase is dried with a desiccant, concentrated, and purified by column chromatography to obtain a compound of formula V; the catalyst used is DMAP, and the condensing agent used is EDC.

[0039] The invention will be better understood through the following description, which is merely illustrative and does not limit the advantageous embodiments of the invention to these examples.

[0040] Example 1

[0041] Succinimidyl-O-methylserine bornyl ester (RJ-1)

[0042]

[0043] Preparation of succinimide-O-methylserine: Place succinimide potassium salt (5.0 g, 50.5 mmol), O-methylserine (6.6 g, 55.5 mmol), and 60 mL of dioxane in a 100 mL reaction bottle, heat to 100°C, react for 6 hours, evaporate all the solvent under reduced pressure, add 20 mL of water to the residue, stir for 20 min, filter, and collect the off-white solid as succinimide-O-methylserine.

[0044] Preparation of succinimidyl-O-methylserine borneol ester: Place succinimidyl-O-methylserine (2.0 g 9.95 mmol) and 80 mL dichloromethane in a 100 mL reaction bottle and dissolve under stirring; add borneol (1.53 g, 9.95 mmol), EDC (2.85 g, 14.92 mmol), DMAP (243 mg, 1.99 mmol), and react at room temperature for 4-6 h until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (50 mL*2), collect the organic phase, wash with 20 mL of water phase and brine once in turn, dry the dichloromethane phase over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate. The product is 2.42 g of colorless oil; 72% yield.

[0045] Example 2

[0046] Succinimidyl-glycine 4-hydroxybenzyl ester (RJ-2)

[0047]

[0048] The preparation method of succinimide-glycine is the same as that of Example 1, except that the raw material is replaced with glycine, and the other operation steps are the same.

[0049] In a 100 ml reaction bottle, add succinimide-glycine (1.56 g 9.95 mmol) and 80 mL dichloromethane, stir and dissolve; add 4-hydroxybenzyl alcohol (1.23 g, 9.95 mmol), EDC (2.85 g, 14.92 mmol), DMAP (243 mg, 1.99 mmol), react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (50 mL*2), collect the organic phase, wash with 20 mL of water and brine once, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% ethyl acetate / petroleum ether-15% methanol / dichloromethane.

[0050] The product is 0.91 g of off-white solid; 42% yield; 1 H NMR (400 MHz, CDCl 3 )δ7.28(d,J=7.2Hz,2H),6.84(d,J=7.2Hz,2H),5.11(s,2H),4.28(s,2H),2.79(s,4H); 13 C NMR (101 MHz, CDCl 3)δ176.3,166.6,158.7,129.4,129.1,115.4,115.0,67.5,39.6,28.2; HRMS(ESI):Exactmass calcd for C 13 H 13 NO 5 ,[M+H] + ,264.0866.Found 264.0860.

[0051] Example 3

[0052] Succinimidyl-isoleucine bornyl ester (RJ-3)

[0053]

[0054] The preparation method of succinimide-isoleucine is the same as that of Example 1, except that the raw material is replaced with isoleucine, and the other operation steps are the same.

[0055] In a 100 ml reaction bottle, add succinimide-isoleucine (1.24 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine once, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.9 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate.

[0056] The product is 1.18 g of colorless oil; 68% yield). Major isomer: minor isomer = 2.0:1.0;

[0057] 1 H NMR (400 MHz, CDCl 3 )δ4.90-4.83(m,1H),4.53(d,J=8.0Hz,1H),2.76(s,4H),2.47-2.32(m,2H),1.73-1.68(m,4H),1.28-1.17(m,3 H),1.11(d,J=6.4Hz,1H),0.97(d,J=7.2Hz,3H),0.88(s,3H),0.86(s,3H),0.82(d,J=6.4Hz,3H),0.76(s,3H); 13 C NMR (101 MHz, CDCl 3)δ176.5,168.7,81.5,56.8,48.7,47.7,44.7,36.4,34.0,27.9,27.5,27.2,19.6,18.8,15.6,13.4,11.3; HRMS(ESI):Exact mass calcd for C 20 H 31 NO 4 ,[M+Na] + ,372.2145.Found 372.2153.

[0058] Example 4

[0059] Succinimidyl-Valine Bornyl Ester (RJ-4)

[0060]

[0061] The preparation method of succinimide-valine is the same as that of Example 1, except that the raw material is replaced with valine, and the other operation steps are the same.

[0062] In a 100 ml reaction bottle, add succinimide-valine (991 mg, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate.

[0063] The product is 1.24 g of colorless oil; 74.2% yield. Major isomer: minor isomer = 3.0:1.0.

[0064] 1 H NMR (400 MHz, CDCl 3 )δ4.91-4.88(m,1H),4.41(d,J=8.8Hz,1H),2.76(s,4H),2.72-2.64(m,1H),2.38-2.31(m,1H),1.77-1.66(m,4H),1.2 8-1.19(m,3H),1.15(d,J=6.8Hz,3H),1.05-0.98(m,1H),0.89(s,3H),0.86(s,3H),0.84(d,J=7.2Hz,3H),0.79(s,3H);13 C NMR (101 MHz, CDCl 3 )δ176.4,168.5,81.4,58.4,48.7,47.7,44.7,36.5,28.0,27.9,27.6,27.2,21.2,19.6,19.4,18.8; HRMS(ESI): Exact mass calcd for C 19 H 29 NO 4 ,[M+Na] + ,358.1989.Found358.1998.

[0065] Example 5

[0066] Succinimidyl-alanine bornyl ester (RJ-5)

[0067]

[0068] The preparation method of succinimide-alanine is the same as that of Example 1, except that the raw material is replaced with alanine, and the remaining steps are the same.

[0069] In a 100 ml reaction bottle, add succinimide-alanine (851 mg, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate.

[0070] The product is 1.18 g of colorless oil; 77.3% yield. Major isomer: minor isomer = 2.0:1.0,

[0071] 1 H NMR (400 MHz, CDCl 3 )δ4.99-4.82(m,2H),2.75(s,4H),2.40-2.32(m,1H),1.76-1.68(m,4),1.3 1-1.17(m,2H),1.03-1.00(m,2H),0.89(s,3H),0.86(s,3H),0.8s(s,1.5H); 13 C NMR (101 MHz, CDCl 3)δ176.1,169.2,81.6,48.8,48.3,47.8,44.8,36.5,28.1,27.9,27.1,19.6,18.8,14.2,13.3; HRMS(ESI): Exact mass calcd for C 17 H 25 NO 4 ,[M+Na] + ,330.1676.Found 330.1685.

[0072] Example 6

[0073] Succinimidyl-norvaline 4-hydroxybenzyl ester (RJ-6)

[0074]

[0075] The preparation method of succinimide-norvaline is the same as that of Example 1, except that the raw material is replaced with norvaline, and the other operation steps are the same.

[0076] In a 100 ml reaction bottle, add succinimide-norvaline (991 mg, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add 4-hydroxybenzyl alcohol (0.617 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine once, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with 50% ethyl acetate / petroleum ether as the eluent.

[0077] The product is 0.53 g of colorless oil; 34.6% yield; 1 H NMR (400 MHz, CDCl 3 )δ7.17(d,J=8.4Hz,2H),6.78(d,J=8.4Hz,2H),5.07(dd,J 1 =12.0Hz,J 2 =20.0Hz,2H),4.72(dd,J 1 =5.6Hz,J 2 =10.4Hz,1H),2.72(s,4H),2.15-2.05(m,2H),1.28-1.21(m,2H),0.90(t,J=7.2Hz,3H); 13 CNMR (101MHz, CDCl3 )δ176.8,169.0,156.1,130.3,127.1,115.4,67.5,52.7,29.7,28.0,19.5,13.3; HRMS(ESI): Exact mass calcd for C 16 H 19 NO 5 ,[M+H] + ,306.1336.Found306.1335.

[0078] Example 7

[0079] Succinimidyl-isoleucyl-4-fluorobenzylamine (RJ-7)

[0080]

[0081] The preparation method of succinimide-isoleucine is the same as that of Example 1, except that the raw material is replaced with isoleucine, and the other operation steps are the same.

[0082] In a 100 ml reaction bottle, add succinimide-isoleucine (1.06 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add EDC (1.43 g, 7.46 mmol), HOBt (673 mg, 4.98 mmol), DMAP (122 mg, 1.0 mmol), add 4-fluorobenzylamine (0.622 g, 4.98 mmol) after 10 min, and react at room temperature for 4-6 hours until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with saturated sodium bicarbonate solution, aqueous phase, and saturated brine in turn, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.8 g of crude product. The crude product is chromatographed on a silica gel column, and the product is 1.15 g of colorless oil; 72.5% yield;

[0083] 1 H NMR (400 MHz, CDCl 3 )δ7.15(dd,J 1 =5.2Hz,J 2 =8.8Hz,2H),6.93(t,J=8.8Hz,2H),4.38(dd,J 1 =6.0Hz,J 2 =14.5Hz,2H),4.32((t,J=11.6Hz,1H),4.21(dd,J 1 =5.6Hz,J 2=14.4Hz,1H),2.67(s,4H),2.53-2.50(m,1H),1.20-1.16(m,1H),0.90(d,J=6.8Hz,3H),0.76(d,J=7.6Hz,3H); 13 C NMR (101 MHz, CDCl 3 )δ177.5,168.4,163.3,1609,133.9,133.8,129.4,129.3,115.6,115.4,62.5,42.8,32.0,25.3,15.6,10.0; 19 F NMR (376 MHz, CDCl 3 )δ-115.0; HRMS(ESI):Exact mass calcd for C 17 H 21 FN 2 O 3 ,[M+H] + ,321.1609.Found321.1601.

[0084] Example 8

[0085] Succinimidyl-valyl benzylamide (RJ-8)

[0086]

[0087] The preparation method of succinimide-valine is the same as that of Example 1, except that the raw material is replaced with valine, and the other operation steps are the same.

[0088] In a 100 ml reaction bottle, add succinimide-valine (0.991 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add EDC (1.43 g, 7.46 mmol), HOBt (673 mg, 4.98 mmol), DMAP (122 mg, 1.0 mmol), add benzylamine (0.622 g, 4.98 mmol) after 10 min, react at room temperature for 4-6 h until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with saturated sodium bicarbonate solution, aqueous phase, and saturated brine in sequence, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.5 g of crude product. The crude product is chromatographed on a silica gel column with 50% ethyl acetate / petroleum ether as the eluent.

[0089] The product is 1.05 g of colorless oil; 73.4% yield; 1 H NMR (400 MHz, CDCl 3)δ7.55-7.26(m,5H),4.52(dd,J 1 =6.4Hz,J 2 =14.8Hz,1H),4.39-433(m,2H),2.83-2.74(m,1H),2.77(s,4H),1.07(d,J=6.4Hz,3H),0.82(d,J=6.4Hz,3H); 13 C NMR (101 MHz, CDCl 3 )δ177.4,168.2,137.9,128.7,128.0,127.5,63.9,43.6,27.9,26.6,19.7,19.3; HRMS(ESI):Exact masscalcd for C 16 H 20 FN 2 O 3 ,[M+H] + ,289.1547.Found 289.1541.

[0090] Example 9

[0091] Succinimidyl-norvaline bornyl ester (RJ-9)

[0092]

[0093] The preparation method of succinimide-norvaline is the same as that of Example 1, except that the raw material is replaced with norvaline, and the other operation steps are the same.

[0094] In a 100 ml reaction bottle, add succinimide-norvaline (991 mg, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water phase and brine once, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate. The product is 1.20 g of colorless oil; 72.6% yield.

[0095] 1 H NMR (400 MHz, CDCl 3 )δ4.90-4.86(m,1H),4.72(dd,J 1 =6.0Hz,J2 =9.6Hz,1H),2.76(s,4H),2.37-2.29(m,1H),2.14-2.09(m,2H),1.74-1.65(m,3H),1.30-1.25(m,4H),0.94(t,J 1 =7.2Hz,3H),0.88(s,3H),0.86(s,3H),0.82(s,3H); 13 C NMR (101 MHz, CDCl 3 )δ176.4,169.0,81.6,52.7,48.7,47.7,44.8,36.2,29.7,28.0,27.8,27.2,19.6,18.8,13.4; HRMS(ESI): Exact mass calcd for C 19 H 29 NO 4 ,[M+Na] + ,358.1989.Found 358.1998.

[0096] Example 10

[0097] Succinimidyl-phenylalanine bornyl ester (RJ-10)

[0098]

[0099] The preparation method of succinimide-phenylalanine is the same as that of Example 1, except that the raw material is replaced with phenylalanine, and the other operation steps are the same.

[0100] In a 100 ml reaction bottle, add succinimide-phenylalanine (1.23 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water phase and brine once, collect the organic phase, dry it over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate. The product is 1.49 g of colorless oil; 78.1% yield.

[0101] 1 H NMR (400 MHz, CDCl 3 )δ7.27-7.13(m,5H),5.03(dd,J 1=5.6Hz,J 2 =11.2Hz,1H),4.97-4.94(m,1H),3.50-3.40(m,2H),2.61-2.49(m,4H),2.41-2.33(m,1H),1.74-1.68(m,3H),1.29-1.21(m,2H),1.04(dd,J 1 =3.2Hz,J 2 =13.6Hz,1H),0.90(s,3H),0.87(s,3H),0.82(s,3H); 13 C NMR (101 MHz, CDCl 3 )δ176.2,168.4,136.6,128.8,128.5,126.9,81.9,53.7,48.9,47.8,44.8,27.1,19.6,18.8,13.4; HRMS(ESI):Exact mass calcd forC 23 H 29 NO 4 ,[M+Na] + ,406.1989.Found 406.1999.

[0102] Embodiment 11

[0103] Succinimidyl-leucine bornyl ester (RJ-11)

[0104]

[0105] The preparation method of succinimide-leucine is the same as that of Example 1, except that the raw material is replaced with leucine, and the other operation steps are the same.

[0106] In a 100 ml reaction bottle, add succinimide-leucine (1.06 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.768 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water phase and brine once, collect the organic phase, dry it over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate. The product is 1.33 g of colorless oil; 76.5% yield.

[0107] 1 H NMR (400 MHz, CDCl3 )δ4.90-4.85(m,1H),4.82-4.77(m,1H),2.74(s,4H),2.37-2.30(m,1H),2.18-2.10(m,1H ),1.95-1.88(m,1H),1.76-1.63(m,3H),1.46-1.39(m,1H),1.32-1.15(m,2H),1.00(dd,J 1 =3.2Hz,J 2 =13.6Hz,1H),0.93(s,3H),0.92(s,3H),0.88(s,3H),0.85(s,3H),0.79(s,2H); 13 C NMR (101 MHz, CDCl 3 )δ176.5,169.3,81.6,51.4,48.8,47.8,44.7,36.6,36.4,28.1,27.9,27.1,25.1,23.0,21.2,19.6,18.8,13.3; HRMS(ESI):Exact mass calcd for C 20 H 31 NO 4 ,[M+Na] + ,372.2145.Found 372.2155.

[0108] Example 12

[0109] Succinimidyl-leucyl-4-fluorobenzylamine (RJ-12)

[0110]

[0111] The preparation method of succinimide-leucine is the same as that of Example 1, except that the raw material is replaced with leucine, and the other operation steps are the same.

[0112] In a 100 ml reaction bottle, add succinimide-leucine (1.06 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add EDC (1.43 g, 7.46 mmol), HOBt (673 mg, 4.98 mmol), DMAP (122 mg, 1.0 mmol), add 4-fluorobenzylamine (0.622 g, 4.98 mmol) after 10 min, and react at room temperature for 4-6 h until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with saturated sodium bicarbonate solution, aqueous phase, and saturated brine in sequence, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.8 g of crude product. The crude product is chromatographed on a silica gel column with 50% ethyl acetate / petroleum ether as the eluent.

[0113] The product is 1.25 g of white solid; 83.1% yield; 1 H NMR (400 MHz, CDCl 3 )δ7.24-7.20(m,2H),7.01(t,J=8.4Hz,1H),6.48(brs,1H),4.80(dd,J 1 =5.2Hz,J 2 =10.8Hz,1H),4.44(m,2H),2.73(s,4H),2.34-2.26(m,2H),1.88-1.82(m,1H),1. 74-1.67(m,2H),1.42-1.35(m,1H),0.92(d,J=8.0Hz,3H),0.88(d,J=8.0Hz,3H); 13 C NMR (101 MHz, CDCl 3 )δ177.1,168.6,163.4,160.9,133.6,133.6,129.4,129.3,115.6,115.4,53.7,43.0,41.9,36.7,28.0,27.0,25.3,23.0,21.3; 19 FNMR (376MHz, CDCl 3 )δ-114.9; HRMS(ESI):Exact mass calcd forC 17 H 21 FN 2 O 3 ,[M+H] + ,321.1609.Found 321.1604.

[0114] Embodiment 13

[0115] Succinimidyl valine 4-hydroxybenzyl ester (RJ-13)

[0116]

[0117] The preparation method of succinimide-valine is the same as that of Example 1, except that the raw material is replaced with valine, and the other operation steps are the same.

[0118] In a 100 ml reaction bottle, add succinimide-valine (991 mg, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add 4-hydroxybenzyl alcohol (0.617 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 2.0 g of crude product. The crude product is chromatographed on a silica gel column with 50% ethyl acetate / petroleum ether as the eluent.

[0119] The product is 0.45 g of colorless oil; 30.3% yield; 1 H NMR (400 MHz, CDCl 3 )δ7.16(d,J=8.0Hz,2H);6.78(d,J=8.0Hz,2H),5.10-5.02(m,2H),4.41(d,J=8.8Hz ,1H),2.70(s,4H),2.72-2.62(m,1H),1.10(d,J=6.4Hz,3H),0.82(d,J=6.4Hz,3H); 13 C NMR (101 MHz, CDCl 3 )δ176.7,168.3,156.0,130.3,115.5,67.1,58.2,27.9,27.7,21.0,19.3; HRMS(ESI):Exactmass calcd for C 16 H 19 NO 5 ,[M+H] + ,306.1336.Found 306.1330.

[0120] Embodiment 14

[0121] Succinimidyl-isoleucine-4-hydroxybenzyl ester (RJ-14)

[0122]

[0123] The preparation method of succinimide-isoleucine is the same as that of Example 1, except that the raw material is replaced with isoleucine, and the other operation steps are the same.

[0124] In a 100 ml reaction bottle, add succinimide-isoleucine (1.06 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add 4-hydroxybenzyl alcohol (0.617 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine once, collect the organic phase, dry over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.5 g of crude product. The crude product is chromatographed on a silica gel column with 50% ethyl acetate / petroleum ether as the eluent.

[0125] The product is 0.57 g of colorless oil; 36.4% yield; 1 H NMR (400 MHz, CDCl 3 )δ7.16(d,J=8.4Hz,2H),6.78(d,J=8.4Hz,2H),5.90(brs,1H),5.09-5.01(m,2H),4.55-4.49(m,1H),2.68 (d,J=3.2Hz,4H),2.45-2.41(m,1H),1.40-1.33(m,1H),1.05(J=6.4Hz,2H),0.92(J=7.2Hz,1H),0.82(dd,J 1 =6.2Hz,J 2 =16.0 Hz, 3H); 13 C NMR (101 MHz, CDCl 3 )δ176.8,168.4,156.1,130.3,127.2,115.4,67.2,57.7,33.1,28.0,25.7,16.9,10.9; HRMS(ESI): Exact masscalcd for C 17 H 21 NO 5 ,[M+Na] + ,342.1317.Found 342.1325.

[0126] Embodiment 15

[0127] Succinimidyl-Isoleucine Vanillyl Ester (RJ-15)

[0128]

[0129] The preparation method of succinimide-isoleucine is the same as that of Example 1, except that the raw material is replaced with isoleucine, and the other operation steps are the same.

[0130] In a 100 ml reaction bottle, add succinimide-isoleucine (1.06 g, 4.98 mmol) and 50 mL of dichloromethane, stir and dissolve; add vanillyl alcohol (0.767 g, 4.98 mmol), EDC (1.43 g, 7.46 mmol), DMAP (122 mg, 1.0 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2.0 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water phase and brine once, collect the organic phase, dry it over anhydrous sodium sulfate, and concentrate it under vacuum to obtain the crude product. The crude product is chromatographed on a silica gel column with 70% ethyl acetate / petroleum ether as the eluent. The product is 0.46 g of colorless oil with a yield of 26.4%.

[0131] Products: Isomers are approximately 2:1; 1 H NMR (400 MHz, CDCl 3 )δ6.90-6.80(m,3H),5.75(s,1H),5.11-4.99(m,2H),4.55-4.48(m,1H),3.87(s,3H),2.70(s, 4H),2.45-2.41(m,1H),1.40-1.33(m,1H),1.05(J=6.4Hz,2H),0.92(J=7.2Hz,1H),0.82(dd,J 1 =6.2Hz,J 2 =16.0 Hz, 3H); 13 CNMR (101MHz, CDCl 3 )δ176.6,168.4,146.5,145.8,127.2,121.9,114.3,111.3,67.4,57.6,55.9,33.8,28.0,25.7,16.8,10.9; HRMS (ESI): Exact mass calcd for C 18 H 23 NO 6 ,[M+H] + ,350.1598.Found 350.1591.

[0132] Example 16

[0133] Succinimidyl-glycine ethyl ester (RJ-16)

[0134]

[0135] In a 100 ml reaction bottle, potassium succinimide (5.0 g, 50.5 mmol), glycine ethyl ester (5.72 g, 55.5 mmol), and 60 mL of dioxane were added, heated to 100 ° C, reacted for 6 hours, and all solvents were evaporated under reduced pressure. 20 mL of water was added to the residue, stirred for 20 minutes, and filtered to collect the off-white solid as succinimide-glycine ethyl ester. The crude product was subjected to silica gel column chromatography, and the eluent was 60% ethyl acetate / petroleum ether. The product was 9.4 g of light yellow solid, with a yield of 90.3%.

[0136] 1 H NMR (400 MHz, CDCl 3 )δ4.24(s,2H),4.20(q,J=7.2Hz,2H),2.79(s,4H),1.27(t,J=7.2Hz,3H); HRMS(ESI): Exact mass calcd for C 8 H 11 NO 4 ,[M+H] + ,186.0671.Found186.0665.

[0137] Embodiment 17

[0138] Succinimidyl-phenylglycine bornyl ester (RJ-17)

[0139]

[0140] The preparation method of succinimide-phenylglycine is the same as that of Example 1, except that the raw material is replaced with phenylglycine, and the other operation steps are the same.

[0141] In a 100 ml reaction bottle, add succinimide-phenylglycine (1.0 g, 4.23 mmol) and 50 mL of dichloromethane, stir and dissolve; add borneol (0.651 g, 4.23 mmol), EDC (1.23 g, 6.44 mmol), DMAP (103 mg, 0.84 mmol), and react at room temperature for 4-6 hours until the raw material disappears. Add 2 mol / L hydrochloric acid to adjust the pH to 4-5, extract with dichloromethane (30 mL*2), collect the organic phase, wash with 20 mL of water and brine once, collect the organic phase, dry it over anhydrous sodium sulfate, and concentrate under vacuum to obtain 1.8 g of crude product. The crude product is chromatographed on a silica gel column with an eluent of 10%-40% dichloromethane / ethyl acetate. The product is 1.25 g of colorless oil; 80.2% yield.

[0142] 1 H NMR (400 MHz, CDCl3 )δ7.51-7.49(m,2H),7.35-7.32(m,3H),5.84(d,J=2.4Hz,1H),4.98-4.91(m,1H),2.74(s,4H),2.42-2.31(m,1H), 1.69-1.62(m,3H),1.49-1.46(m,1H),1.24-1.03(m,3H),0.88(s,3H),0.84(s,3H),0.82(s,1.5H),0.77(s,1.5H); 13 C NMR (101 MHz, CDCl 3 )δ175.8,167.6,134.1,130.0,128.6,128.4,82.3,56.6,48.9,47.0,44.8,36.6,36.2,28.2,27.8,27.0,19.6,18.8,13.4; HRMS (ESI): Exact mass calcd forC 22 H 27 NO 4 ,[M+Na] + ,392.1832.Found 392.1841.

[0143] Study on the biological activities of succinimide derivatives

[0144] Succinimide derivatives are used to treat convulsions and epilepsy. This project uses the maximum electroconvulsive test (MES) to further study and evaluate the anti-epileptic activity of the drug, uses neurotoxicity experiments to evaluate the safety of the compound, uses the classic chemical model of pentylenetetrazol to study and evaluate the anticonvulsant pharmacological mechanism of the drug, uses the elevated plus maze test, open field test and fear box to examine the anxiety-like behavior and suggestive conditioned memory behavior of mice, uses HE staining technology to observe the growth of hippocampal neurons and their morphological changes in epileptic mice after administration, and conducts hepatotoxicity tests to examine the effects of the compound on mouse liver function.

[0145] (1) Maximum electroconvulsive test (MES) and neurotoxicity test

[0146] MES method: The mice are preliminarily screened before the experiment, and only qualified mice can be used for the next experiment. The screening method is as follows: One day before the experiment, the experimental mice are subjected to 50V, 50Hz electrical stimulation, two electrodes are clamped on the ears of the mice, and the electricity is stimulated for 0.2s. The mice with hind limb rigidity are reserved for the experiment and used for later experiments. The method for determining the maximum electroconvulsive test is as follows: After the test compound is dissolved, it is administered by gavage. 0.5h after administration, the ear electrodes are stimulated for 0.2s, 50V, 50Hz. If the mice do not have hind limb rigidity, it indicates that the test compound has anticonvulsant activity at this dose.

[0147] Neurotoxicity test method: Before the experiment, the mice were placed on an XZC-4B rotating rod with a radius of 0.4 cm and a rotation speed of 24 rap / min for training. The training was continued for 2 to 3 days, 1 to 3 times a day, to fully ensure that the mice used in this model could adapt to the experimental conditions before administration. Mice that did not fall from the fatigue meter within 3 minutes or fell less than 3 times were selected as subjects for the neurotoxicity experiment. Then the qualified mice were randomly divided into groups and a double-blind administration method was adopted. The mice in the administration group were intraperitoneally injected with 100 mg / kg of the test compound, and the mice were placed steadily on the fatigue meter at 0.5, 1 and 2 hours after administration, respectively, and rotated at 24 rap / min. If the test mice did not fall or fell less than 3 times within 3 minutes, it indicated that the compound had no neurotoxicity at this dose; if the number of falls exceeded 3 times or more, it indicated that the test compound had neurotoxicity at this dose. The number of mice with neurotoxic reactions was recorded to preliminarily evaluate the safety of the compound.

[0148] In the MES model, the antiepileptic activity of the compound was preliminarily evaluated 0.5 h after intraperitoneal administration; in the neurotoxicity model, the neurotoxicity of the compound was evaluated 0.5 h after intraperitoneal administration (Table 1). As shown in Table 1, the compound did not show neurotoxicity at the experimental dose.

[0149] Table 1 Results of MES and neurotoxicity experiments of succinimide derivatives (n=3)

[0150]

[0151]

[0152] a Number of mice without compulsive seizures / number of experimental mice;

[0153] b MES detection was performed 0.5 h after intraperitoneal injection of each compound;

[0154] c Number of mice without neurotoxicity / number of experimental mice;

[0155] In the MES model, the antiepileptic activity of the compounds was preliminarily evaluated 0.5 h after intraperitoneal administration. As shown in Table 1, succinimide derivatives can protect mice from rigidity under stimulation and have certain antiepileptic activity. In the neurotoxicity model, the neurotoxicity of the compounds was evaluated 0.5 h after intraperitoneal administration. As shown in Table 1, the compounds showed no neurotoxicity at the experimental doses.

[0156] (2) Experimental acute epilepsy model induced by pentylenetetrazol

[0157] Method 1: Mice were randomly divided into a drug-treated group (50 mg / kg), a positive control group and a model group, with 3 mice in each group. The drug-treated group was intraperitoneally injected with the test compound, the control group was intraperitoneally injected with the positive drug sodium valproate, and the model group was gavaged with normal saline. 30 minutes later, the mice were subcutaneously injected with 85 mg / kg pentylenetetrazol. The test animals were placed individually in a mouse cage for observation for 30 minutes. The latency time of clonic seizure, seizure grade, number of clonic seizures, number of tonic seizures and number of deaths of each group of mice were recorded (Table 2).

[0158] Table 2 Effects of succinimide derivatives on acute epileptic seizures induced by PTZ (85 mg / kg) in mice (n=3)

[0159]

[0160]

[0161] Data are expressed as mean ± SD;

[0162] a Number of mice with generalized tonic-clonic seizures / number of tested mice;

[0163] b Number of mice with clonus / number of tested mice;

[0164] c Number of dead mice / number of tested mice;

[0165] The PTZ dose in the model group and the fourteen compound groups was 85 mg / kg.

[0166] Method 2: The mice were randomly divided into a drug-treated group (25 mg / kg, 75 mg / kg), a positive control group (sodium valproate, VPA) and a model group, with 3 mice in each group. The drug-treated group was intraperitoneally injected with the test compound, the control group was intraperitoneally injected with the positive drug VPA, and the model group was gavaged with normal saline. After 30 minutes, the mice were subcutaneously injected with 85 mg / kg of pentylenetetrazol. The test animals were placed individually in a mouse cage for observation for 30 minutes. The latency time of clonic seizure, seizure grade, number of clonic seizures, number of tonic seizures and number of deaths of each group of mice were recorded (Table 3).

[0167] Table 3 Effects of succinimide derivatives on acute epileptic seizures induced by PTZ (85 mg / kg) in mice (n=3)

[0168]

[0169]

[0170] Data are expressed as mean ± SD;

[0171] a Number of mice with generalized tonic-clonic seizures / number of tested mice;

[0172] b Number of mice with clonus / number of mice tested;

[0173] c Number of dead mice / number of tested mice.

[0174] According to Table 2, in the acute convulsion model test against pentylenetetrazol chemical drugs, compared with the model group, most succinimide derivatives at the same dose can prolong the duration of clonic seizures in mice, reduce the grade of clonic seizures in mice, inhibit tonic seizures in mice, and reduce the mortality rate of mice. According to Table 3, most succinimide derivatives at different doses can also prolong the duration of clonic seizures in mice, reduce the grade of clonic seizures in mice, inhibit tonic seizures in mice, and reduce the mortality rate of mice. This indicates that succinimide derivatives may have the effect of inhibiting or reducing convulsions in mice induced by pentylenetetrazol.

[0175] (3) Pentylenetetrazol-induced chronic epilepsy model in mice

[0176] Methods: The experimental mice were divided into a blank group, a positive drug (VPA) and an epilepsy group. The blank group was given an equal volume of normal saline. The mice in the epilepsy group were intraperitoneally injected with PTZ (35 mg / kg) once every other day for 10 consecutive times. The seizures of the mice were observed and recorded for 30 minutes. After three consecutive subcutaneous injections of PTZ, the mice showed three consecutive seizures, which was a successful ignition, and they could be used in later experiments (Table 4). The mice with successful ignition were randomly divided into a drug administration group (25 mg / kg, 50 mg / kg, 75 mg / kg), a positive control group and a model group, with 6 mice in each group. The drug administration group was intraperitoneally injected with the test compound, the positive control group was intraperitoneally injected with the positive drug sodium valproate, and the model group was gavaged with normal saline. After 30 minutes, the mice were subcutaneously injected with PTZ (35 mg / kg), and the test animals were placed in a mouse cage for observation for 30 minutes. The latency time of clonic seizures, the number of clonic seizures, the number of tonic seizures and the number of deaths of each group of mice were recorded (Table 5).

[0177] Table 4 Effects of PTZ (35 mg / kg) on ​​chronic epileptic seizures in mice (n=6)

[0178]

[0179] Data are expressed as mean ± SD.

[0180] Table 5 Effects of succinimide derivatives on chronic epileptic seizures in mice induced by PTZ (35 mg / kg) (n=6)

[0181]

[0182] Data are expressed as mean ± SD;

[0183] a Number of mice with generalized tonic-clonic seizures / number of tested mice;

[0184] b Number of mice with clonus / number of tested mice;

[0185] c Number of dead mice / number of tested mice.

[0186] During the pentylenetetrazol modeling process, the duration of clonic seizures in mice gradually shortened until the modeling was successful. In the test of the convulsion model induced by pentylenetetrazol chemical drugs, the duration of clonic seizures in mice in the positive drug group was significantly prolonged compared with the model group, and the grade of clonic seizures in mice was reduced. Compared with the model group, the duration of clonic seizures in mice in the low, medium and high dose groups of succinimidyl-norvaline bornyl ester (RJ-9) was prolonged. Compared with the model group, the duration of clonic seizures in mice in the high dose group of succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14) was prolonged.

[0187] (4) Pentylenetetrazol-induced zebrafish epilepsy model

[0188] The experimental group selected zebrafish larvae, the model group was given PTZ; the positive drug group mice were given VPA (500μM) + PTZ; the drug groups were given the compound succinimidyl-norvaline bornyl ester (RJ-9) and the compound succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14), respectively. After drug administration, the swimming behavior of zebrafish in each group was detected by ZebraBox, and the total swimming distance and swimming speed were calculated. The effects of the compounds on the epileptic behavior of zebrafish were evaluated.

[0189] In the experiment, four groups of zebrafish larvae treated with drugs the day before and one group of zebrafish not treated with drugs were transferred to 15mmol PTZ, and then immediately placed in a 48-well plate, with one fish in each well. The 48-well plate was placed in a zebrafish behavior trajectory tracking system. After standing in the dark for about 10 minutes, the zebrafish behavior was recorded for 30 minutes, and data was collected every 5 minutes to track and record the behavior trajectory of each group. Measurements using a zebrafish behavior detector found that the compounds succinimidyl-norvaline borneol ester (RJ-9) and succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14) did not produce toxic reactions to zebrafish at the experimental doses ( Figure 1 ).

[0190] The study showed that the total movement distance of the larvae in the PTZ model group increased, and the movement trajectory diagram showed an overexcited state. Compounds RJ-9 and RJ-14 at 50 μM can significantly reduce the movement distance of zebrafish epilepsy symptoms, showing significant anti-epileptic activity ( Figure 2 ).

[0191] (5) Elevated plus maze (EPMT) test

[0192] On the 7th day after administration, EPMT was used to test epileptic mice behavior in a room where sound and light could be controlled. The elevated plus maze is composed of two opposite equal-length open arms and closed arms connected by a central area. The length of the four arms is 50 cm × 10 cm, and the size of the central area is 10 cm × 10 cm. The material is blue high-grade medical organic board, and the bottom of the maze is 50 cm from the ground. At the beginning of the experiment, the mouse was placed in the central area with its head facing away from the person and placed in the maze facing the open and closed arms. The head and forelimbs of the mouse were considered to enter the arm, and those outside the arm were considered to exit. After each mouse experiment, the four arms were carefully wiped with 75% ethanol and the mouse excrement was cleaned to avoid affecting the next experiment. The number of times the mouse entered the open arm and the retention time within 10 minutes were analyzed by video on the computer, and the corresponding percentage was calculated to reflect the anxiety-like behavior of the mouse.

[0193]

[0194] Table 6 Ratio of time and number of times each group of mice entered the elevated plus maze open arm on the 7th day of administration

[0195]

[0196]

[0197] Data are expressed as mean ± SD.

[0198] In the elevated plus maze (EPMT) test, as shown in Table 6, on the 7th day of administration, compared with the model group, the time ratio of mice entering the open arms of the elevated plus maze in the positive drug group and the low, medium and high dose groups of succinimide-norvaline bornyl ester (RJ-9) and the low and medium dose groups of succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) increased. Compared with the model group, the number of times mice entering the open arms of the elevated plus maze in the positive drug group and the low, medium and high dose groups of succinimide-norvaline bornyl ester (RJ-9) and the low and high dose groups of succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) increased. The results show that succinimide derivatives have the effect of improving the anxiety-like behavior of PTZ-induced epilepsy model mice.

[0199] (6) Open field (OFT) test

[0200] The open field test (OFT) was performed on the 7th day after administration. In this study, the open field test box was divided into nine equal-sized grids. After the mice were placed in the central area, the experimental operator immediately left the open field, and the entire experiment was completed in a ventilated, quiet, and dimly lit space. A video analysis system was used to record the movement distance and retention time of the mice in the central area for 5 minutes to reflect the anxiety-like behavior of the mice. After the experiment on each mouse, the experimental instruments were cleaned with 75% ethanol and the mouse excrement was removed.

[0201] Table 7 Movement distance and retention time of mice in each group in the central area of ​​the open field on the 7th day of administration

[0202]

[0203] Data are expressed as mean ± SD;

[0204] * Indicates significant difference from the model group (P<0.05);

[0205] ## It indicates significant difference compared with the blank group (P<0.01).

[0206] Mice with lower anxiety tend to move longer distances and stay longer in the center of the open field, i.e., the open area. As shown in Table 7, on the 7th day of administration, the movement distance of mice in the model group in the central area of ​​the open field decreased compared with the blank group (P<0.01); compared with the model group, positive drugs and low, medium, and high doses of succinimide derivatives can increase the movement distance of PTZ-induced epileptic mice in the central area of ​​the open field, among which 50 mg / kg succinimide-norvaline bornyl ester (RJ-9) and 25 mg / kg succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) can significantly increase the movement distance of PTZ-induced epileptic mice in the central area of ​​the open field (P<0.05), and the difference is statistically significant.

[0207] As shown in Table 7, on the 7th day of administration, compared with the blank group, the model group mice's retention time in the central area of ​​the open field was significantly reduced (P<0.05), and the difference was statistically significant; compared with the model group, positive drugs and low, medium, and high doses of succinimide derivatives can increase the retention time of PTZ-induced epileptic mice in the central area of ​​the open field, among which the high dose of succinimide-norvaline bornyl ester (RJ-9) can significantly increase the retention time of PTZ-induced epileptic mice in the central area of ​​the open field (P<0.05), and the difference is statistically significant. The results show that succinimide derivatives can improve the anxiety-like behavior of PTZ-induced epileptic model mice, improve the motor ability of mice, and increase the exploratory behavior of mice.

[0208] (7) Fear Box (FCT) Experiment

[0209] The experiment was divided into 2 phases:

[0210] ① Training period: The mice were placed on the partition of the electric shock generator in the fear box to adapt for 3 minutes, and the electric shock size was turned on with 0.35mA, an interval of 62s, a duration of 2s, and a cycle of 3 times. ② Testing period: The mice were placed in the fear box, the environment was consistent with the training stage, and they were adapted for 3 minutes without stimulation. The video analysis system was used to analyze the freezing time and number of mice in the fear box for 6 minutes. After the experiment of each mouse, the mouse excrement was removed, the venue was wiped with 75% ethanol, and it was fully dried before subsequent experiments.

[0211] Table 8 The duration and number of freezing in the fear box during the training period of each group of mice on the 7th day of drug administration

[0212]

[0213]

[0214] Data are expressed as mean ± SD;

[0215] *Indicates significant difference from the model group (P<0.05); ** Indicates significant difference from the model group (P<0.01); *** Indicates significant difference from the model group (P < 0.001);

[0216] ## It indicates significant difference compared with the blank group (P<0.01). ### It indicates that the difference is significant compared with the blank group (P<0.001).

[0217] As shown in Table 8, during the training period of the fear box (FCT) experiment, compared with the model group, the freezing time of PTZ-induced epileptic mice in the positive drug group, the low, medium and high dose groups of succinimide-norvaline bornyl ester (RJ-9), and the low and medium dose groups of succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) increased in the fear box, among which the high dose group of succinimide-norvaline bornyl ester (RJ-9) had a significant effect on increasing the freezing time of PTZ-induced epileptic mice in the fear box (P<0.0001), and the difference was statistically significant. The freezing times of PTZ-induced epileptic mice in the positive drug group and the low, medium and high dose groups of succinimide derivatives in the fear box increased.

[0218] During the FCT experiment, compared with the blank group, the immobility time of the model group mice in the fear box was significantly reduced (P<0.01), and the number of immobility times was significantly reduced (P<0.0001). Compared with the model group, the immobility time and number of immobility times of the PTZ-induced epileptic mice in the positive drug group and the low, medium and high dose groups of succinimide derivatives in the fear box were increased, among which the immobility time of the PTZ-induced epileptic mice in the positive drug group and the medium dose group of succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) in the fear box was significantly increased (P<0.0001); the immobility time of the PTZ-induced epileptic mice in the medium dose group of succinimide-norvaline bornyl ester (RJ-9) and the high dose group of succinimide-isoleucine 4-hydroxybenzyl ester (RJ-14) in the fear box was significantly increased (P<0.01), and the difference was statistically significant. The positive drug group, the low- and medium-dose groups of succinimidyl-norvaline bornyl ester (RJ-9), and the medium- and high-dose groups of succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14) significantly increased the number of freezing times of PTZ-induced epileptic mice in the fear box (P < 0.0001); the high-dose group of succinimidyl-norvaline bornyl ester (RJ-9) significantly increased the number of freezing times of PTZ-induced epileptic mice in the fear box (P < 0.01), and the difference was statistically significant.

[0219] There are many brain nuclei involved in fear memory, among which scene fear memory is related to the activity of mouse hippocampal neurons. For PTZ-induced epileptic mice, epileptic seizures can cause damage to the mouse hippocampus and neuronal apoptosis. The fear box experiment was used to determine the establishment and extinction of scene memory in PTZ-induced epileptic mice, which is conducive to exploring the protective effect of succinimide derivatives on hippocampal neurons in PTZ-induced epileptic mice.

[0220] As shown in Table 8, compared with the blank group, the model group had a reduced freezing time and frequency in PTZ-induced epilepsy mice, indicating that the episodic fear memory of the mice was extinguished, which may be caused by the damage of neurons in PTZ-induced epilepsy mice. Compared with the low-, medium-, and high-dose groups of succinimide derivatives, the model group was continuously treated with succinimide derivatives, and the freezing time and frequency of the mice increased, indicating that succinimide derivatives can improve the episodic fear memory impairment of PTZ-induced epilepsy model mice, indicating that they have a certain protective effect on hippocampal neurons.

[0221] (8) HE staining results of mouse hippocampus

[0222] HE staining, 200×, succinimidyl-norvaline bornyl ester (RJ-9), succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14), Figure 3 (Left 25 mg / kg, middle 50 mg / kg, right 75 mg / kg, left, middle, right represent the left, middle, and right positions respectively).

[0223] Depend on Figure 3 It can be seen that compared with the blank group, the neuronal apoptosis in the CA1 region of the hippocampal tissue of the model group mice was more serious. Compared with the model group, positive drugs and low, medium and high doses of succinimide derivatives can reduce the apoptosis of neurons in the CA1 region of the hippocampal tissue of mice, and the reduction of neuronal apoptosis in the CA1 region of the hippocampal tissue of mice in the positive drug group was more significant. This shows that succinimide derivatives can improve the apoptosis of neurons in the CA1 region of the hippocampal tissue of PTZ-induced epilepsy model mice.

[0224] (9) Compound hepatotoxicity test

[0225] ALT and AST are mainly present in liver cells. When liver cells are damaged or destroyed, these enzymes will leak from the cells into the blood, causing their concentrations in the blood to increase. Serum ALT and AST are early markers of acute liver injury, with high specificity and sensitivity. They are most commonly used clinically to reflect the degree of liver function damage, and their content is proportional to the degree of liver damage. Therefore, this experiment detected liver ALT and AST after the mice were given the drug. The detection method is as follows: After the experiment, the eyeballs were removed for blood collection, and after being stored in a 4°C refrigerator for 2 hours, they were placed in a 4°C centrifuge at a speed of 3000r / min (centrifugal radius 60mm), and low-temperature centrifugation was performed for 5 minutes to obtain serum, and the content of ALT and AST in the serum was identified using a multifunctional microplate reader. The specific method shall be subject to the instructions in the alanine aminotransferase (ALT) kit and the aspartate aminotransferase (AST) kit.

[0226] As shown in Table 9, the average ALT and AST levels in the model group were higher than those in the blank group. The results showed that the compounds succinimidyl-norvaline bornyl ester (RJ-9) and succinimidyl-isoleucine 4-hydroxybenzyl ester (RJ-14) had no significant liver toxicity.

[0227] Table 9 Effects of succinimide derivatives on mouse liver function

[0228]

[0229] In summary, the study found that succinimide derivatives are relatively safe and have significant anticonvulsant, anti-epileptic, anti-neuropathic pain, anti-depression, neuroprotective, brain damage protection, memory improvement and anti-anxiety activities, and are worthy of further development and use.

[0230] The above is only an embodiment of the present invention, and the common knowledge such as the known specific structure and characteristics in the scheme is not described in detail here. It should be pointed out that for those skilled in the art, several deformations and improvements can be made without departing from the structure of the present invention, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A succinimide derivative, characterized in that: Its general structural formula is shown in Formula V: Wherein: R1 is H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, branched chain, straight chain C1-8 alkyl, cycloalkyl C m H 2m-1 (m=3-6), methylcycloalkyl C m+1 H 2m+1 (m=3-6), substituted allyl, substituted phenyl, substituted benzyl, heterocyclic, methoxymethyl, hydroxymethyl, 2-hydroxyethyl, methylthioethyl, heterocyclic; R2 is methyl, ethyl, isopropyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl branched, straight chain C1-8 alkyl, cycloalkyl C m H 2m-1 (m=3-6), methylcycloalkyl C m+1 H 2m+1 CH2 (m = 3-6), substituted allyl, substituted cinnamyl, rac-2-bornyl, (-)-2-bornyl, (+)-2-bornyl, rac-menthyl, (-)-menthyl, (+)-menthyl, substituted phenyl, substituted benzyl, heterocyclic group, 2-methylpiperidine, dimethylamino, diethylamino, diisopropylamino, 2,2-dimethyl-3-(propylamino)propyl-1-yl, N,2-dimethyl-1-phenylpropyl-2-amino; R3 is H, methyl, ethyl, phenyl; R4 is H, methyl, ethyl, phenyl, spirocyclopentyl; or R3R4 constitutes cyclohexyl, phenyl, (1R,4S)-bicyclo[2,2,1]hexane-2-enyl; X is O, NH or piperazinyl; n is 0 to 3.

2. The succinimide derivative according to claim 1, characterized in that: The R1 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, 2-methyl-n-butyl, neopentyl, n-hexyl, 2-methyl-n-pentyl, 3-methylpentyl, 4-methylpentyl, 2-hexyl, 4-methylpentane-2-yl, 3-methylpentane-2-yl, 3-pentyl, n-heptyl, n-octyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropanemethyl, cyclobutanemethyl, cyclopentanemethyl, cyclohexanemethyl, allyl, 3-methyl-2-buten-1-yl, substituted phenyl, substituted benzyl, methoxymethyl, hydroxymethyl, 2-hydroxyethyl, methylthioethyl.

3. The method for preparing the succinimide derivatives according to claim 1 or 2, characterized in that: The succinimide derivative V is obtained by reacting a compound of the general formula III with a compound of the general formula IV in the presence of a condensing agent, and the reaction formula is as follows:

4. The method for preparing succinimide derivatives according to claim 3, characterized in that: When preparing succinimide derivatives V: add the compound of formula III and the compound of formula IV, a condensing agent, and a catalyst to a reaction container in sequence for reaction, the molar ratio of the compound of formula III to the compound of formula IV is 1.0:1.0-1.5, and react at 5°C-40°C until the compound of formula III and / or the compound of formula IV are consumed; add pure water for washing, extract with ethyl acetate, and concentrate under vacuum to remove ethyl acetate. The organic phase is dried, concentrated, and purified by column chromatography.

5. The method for preparing succinimide derivatives according to claim 4, characterized in that: The condensing agent is DCC, DIC, EDC, BOP, PyBOP, HATU, HBTU, TBTU, ethyl chloroformate, phenyl chloroformate, isopropyl chloroformate or 1-n-propyl phosphoric anhydride.

6. The method for preparing succinimide derivatives according to claim 4, characterized in that: The catalyst is: Et3N, DIEA, DMAP, DPPY, 2,6-lutidine, DABCO or DBU.

7. The method for preparing a succinimide derivative according to any one of claims 4 to 6, characterized in that: The compound of general formula III is obtained by reacting the compound of general formula I with the compound of general formula II, and the reaction formula is shown below:

8. The method for preparing succinimide derivatives according to claim 7, characterized in that: The specific preparation process of the compound of formula III is as follows: add the compound of formula I and the compound of formula II in a molar ratio of 1.0 to 2.0:1.0 to a reaction container in sequence, react at 70-100° C. until the compound of formula I and / or the compound of formula II disappear; concentrate under vacuum to remove the solvent, add water to the residue to precipitate, filter, and dry at 30-60° C. to obtain the compound of formula III.

9. The racemate or optical isomer of the succinimide derivative according to claim 1 or a mixture of its different stereoisomers.

10. Use of the racemic form or optical isomer of the succinimide derivatives according to claim 1 and / or the mixture of different stereoisomers of the succinimide derivatives according to claim 9 in the preparation of sedative, tranquilizing, intellectual, anticonvulsant, anti-epileptic, anti-neuropathic pain, antianxiety, anti-depression, anti-dementia and / or brain damage protective drugs.

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