A 1-(cyclobutylylidene methyl)-2,4,5-trimethoxy benzene compound, and a preparation method and application thereof

Abstract

The application provides a 1-(cyclobutylidene methyl)-2,4,5-trimethoxy benzene compound and a preparation method and application thereof, and belongs to the technical field of antiepileptic drugs. The 1-(cyclobutylidene methyl)-2,4,5-trimethoxy benzene compound prepared by the application can be added with pharmaceutically acceptable excipients to form different dosage forms, and can play the following roles: (1) prolonging the latent period of epileptic clonic seizure and tetanic seizure caused by pentylenetetrazole, reducing the grade and incidence of epileptic seizure, and the effect has a dose-dependent relationship; (2) significantly reducing the grade of acute phase of a rat epilepsy model caused by lithium-piroctone, and improving the survival rate of the rat, and the effect is better than that of a broad-spectrum antiepileptic drug, i.e. sodium valproate.

Description

Technical Field

[0001] This invention relates to the field of antiepileptic drug technology, and in particular to a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound, its preparation method, and its application. Background Technology

[0002] Epilepsy is a common neurological disorder characterized by spontaneous, recurrent seizures caused by abnormal electrical activity in the brain, leading to neuronal damage and even death, as well as alterations in neural networks. The pathogenesis of epilepsy includes an imbalance of excitatory and inhibitory neurotransmitters, and changes in the expression of neurotransmitter-activated receptors and ion channels. Based on clinical seizure type, epileptic seizures can be classified into generalized tonic-clonic seizures (grand mal seizures), absence seizures (petit mal seizures), partial seizures, special seizures, and status epilepticus. There are approximately 65 million people with epilepsy worldwide, including about 10 million in China. Drug therapy remains the preferred treatment method.

[0003] Most currently marketed and investigational antiepileptic drugs primarily target ion channels, exerting their pharmacological effects by reducing the excitability of the nervous system. Their mechanisms of action include: 1) Inhibition of voltage-gated sodium ion channels: Voltage-gated sodium ion channels are the main ion channels constituting the rapid depolarization branch of neuronal action potentials. Inhibiting sodium ion channel currents can inhibit the generation and transmission of action potentials; representative drugs are carbamazepine and phenytoin sodium. 2) Enhancement of GABA. A Receptor activity: GABA A The receptor is activated by the endogenous neurotransmitter GABA, producing inhibitory Cl... － Current enhances GABA A The activity of receptors can reduce cell membrane potential and decrease neuronal excitability; these are benzodiazepines. Targets of antiepileptic drugs: 3) Activation of voltage-gated potassium channels: After potassium channels open, potassium ions flow out in large quantities, and the cell membrane potential decreases. Retegafur, developed by GlaxoSmithKline, is the first potassium channel opener for the treatment of epilepsy. 4) Inhibition of ionotropic glutamate receptors: Glutamate is an important neurotransmitter in the central nervous system. Ionotropic glutamate receptors are coupled with cation channels and are divided into three types: NMDA, KA, and AMPA. Glutamate-mediated neuronal overexcitation plays a key role in inducing epileptic seizures, so inhibiting ionotropic glutamate receptors can treat epilepsy. Representative drugs include the NMDA receptor antagonist ketamine and the AMPA receptor antagonist perampanel. 5) Inhibition of voltage-gated calcium channels: Voltage-gated calcium channels can be divided into L-type, T-type, N-type, R-type, and P / Q-type calcium channels according to their activation voltage and current characteristics. The antiepileptic drugs gabapentin and pregabalin are N-type calcium channel inhibitors, while ethosuximide is a T-type calcium channel inhibitor.

[0004] Sodium valproate is a commonly used broad-spectrum antiepileptic drug in clinical practice, used to treat generalized and partial seizures in patients with epilepsy. Recent studies have shown that sodium valproate can also improve the control rate of status epilepticus. The mechanism of action of sodium valproate is mainly through increasing the concentration of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), thereby enhancing postsynaptic inhibition of neurons and achieving an antiepileptic effect. Furthermore, although there are dozens of commercially available antiepileptic drugs, approximately 30% of patients cannot control their epilepsy with existing medications due to side effects, specific reactions, or drug tolerance. Therefore, the development of novel, highly effective, and low-toxicity antiepileptic drugs remains of significant clinical value and importance. Summary of the Invention

[0005] The purpose of this invention is to provide a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound, its preparation method, and its application, in order to solve the technical problem that commercially available antiepileptic drugs in the prior art cannot effectively control the condition.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene compound having the structure shown in formula (Ⅰ):

[0008]

[0009] This invention provides a method for preparing a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound, comprising the following steps:

[0010] (1) Compound 1a was mixed with dichloromethane, and then compound 1b was added to react and compound 1c was obtained.

[0011] (2) Compound 1c, compound 1d, and aluminum trichloride were mixed with dichloromethane in sequence and reacted to obtain compound 1e;

[0012] (3) Compound 1e, sodium borohydride solution, and sodium hydroxide solution were mixed with tetrahydrofuran in sequence and reacted to obtain compound 1f;

[0013] (4) Compound 1f, sodium acetate and acetic anhydride were mixed and reacted to obtain 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound;

[0014] The compound 1a is The compound 1b is The compound 1d is

[0015] Preferably, in step (1), the mass-to-volume ratio of compound 1a, dichloromethane, and compound 1b is 23–27 g: 130–170 mL: 93–97 g.

[0016] Preferably, in step (1), the reaction temperature is 20-30°C, and the reaction is carried out under stirring conditions until no more bubbles are generated under stirring conditions and the reaction ends.

[0017] Preferably, in step (2), the mass-to-volume ratio of compound 1c, compound 1d, aluminum trichloride, and dichloromethane is 22-27g: 30-33g: 26-30g: 80-120mL;

[0018] The compound 1c is

[0019] Preferably, in step (2), the reaction temperature is 20-30°C and the reaction time is 2-3 hours.

[0020] Preferably, in step (3), the mass-to-volume ratio of compound 1e, sodium borohydride solution, and tetrahydrofuran is 40-45 g: 18-22 mL: 140-160 mL, wherein the concentration of sodium borohydride solution is 0.8-1.0 g / mL.

[0021] The compound 1e is

[0022] Preferably, in step (3), the reaction temperature is 50-65°C and the reaction time is 2-4 hours.

[0023] Preferably, in step (4), the mass-to-volume ratio of compound 1f, sodium acetate, and acetic anhydride is 38-45g: 7-10g: 200-230mL;

[0024] The compound 1f is

[0025] The reaction temperature is 120–140°C, and the reaction time is 2–4 hours.

[0026] This invention provides the application of 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound in the preparation of antiepileptic drugs.

[0027] The 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound provided by this invention can be formulated into different dosage forms with pharmaceutically acceptable excipients. It can be formulated into liquid dosage forms such as emulsion injections and oral emulsions. Those skilled in the art can also prepare it into solid dosage forms such as tablets and capsules using conventional techniques. When the dosage forms are used, they can be administered via intravenous injection, intramuscular injection, or oral administration, as long as the compound with the structure shown in formula (I) of this invention reaches the site of action and achieves an effective concentration.

[0028] The beneficial effects of this invention are:

[0029] (1) The 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound provided by the present invention can reduce the severity and incidence of epileptic seizures caused by pentylenetetrazol, and can also significantly prolong the latency of clonic and tonic seizures, and the effect is dose-dependent.

[0030] (2) The 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound provided by the present invention can significantly reduce the seizure severity in the acute phase of the lithium-pilocarpine-induced rat epilepsy model, improve the survival rate of rats, and has a better effect than the broad-spectrum antiepileptic drug sodium valproate. Attached Figure Description

[0031] Figure 1 This figure shows the survival status of SE model rats after acute phase drug intervention. Detailed Implementation

[0032] This invention provides a 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene compound having the structure shown in formula (Ⅰ):

[0033]

[0034] This invention provides a method for preparing a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound, comprising the following steps:

[0035] (1) Compound 1a was mixed with dichloromethane, and then compound 1b was added to react and compound 1c was obtained.

[0036] (2) Compound 1c, compound 1d, and aluminum trichloride were mixed with dichloromethane in sequence and reacted to obtain compound 1e;

[0037] (3) Compound 1e, sodium borohydride solution, and sodium hydroxide solution were mixed with tetrahydrofuran in sequence and reacted to obtain compound 1f;

[0038] (4) Compound 1f, sodium acetate and acetic anhydride were mixed and reacted to obtain 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound;

[0039] The compound 1a is The compound 1b is The compound 1d is

[0040] In this invention, in step (1), the mass-to-volume ratio of compound 1a, dichloromethane and compound 1b is 23-27g:130-170mL:93-97g, preferably 24-26g:140-160mL:94-96g, and more preferably 25g:150mL:94.5-95.5g.

[0041] In this invention, the mixing of compound 1a with dichloromethane is preferably carried out under nitrogen protection; the mixing of compound 1b with dichloromethane is preferably carried out dropwise.

[0042] In this invention, in step (1), the reaction temperature is 20-30°C, preferably 22-28°C, and more preferably 25°C; the reaction is carried out under stirring conditions until no bubbles are generated under stirring conditions and the reaction ends.

[0043] In this invention, in step (2), the mass-to-volume ratio of compound 1c, compound 1d, aluminum trichloride, and dichloromethane is 22-27g:30-33g:26-30g:80-120mL, preferably 23-26g:30.5-32.5g:27-29g:85-105mL, and more preferably 23.5-25.5g:30.7-32.3g:27.5-28.5g:90-100mL;

[0044] The compound 1c is

[0045] In this invention, the mixing temperature of aluminum trichloride is -2 to 2°C, preferably 0°C.

[0046] In this invention, in step (2), the reaction temperature is 20-30°C, preferably 22-28°C, and more preferably 25°C; the reaction time is 2-3 hours, preferably 2.5 hours.

[0047] In this invention, in step (3), the mass-to-volume ratio of compound 1e, sodium borohydride solution, and tetrahydrofuran is 40-45g:18-22mL:140-160mL, preferably 41-44g:19-21mL:145-155mL, and more preferably 42-43g:20mL:150mL; wherein the concentration of sodium borohydride solution is 0.8-1.0g / mL, preferably 0.85-0.95g / mL, and more preferably 0.87-0.93g / mL;

[0048] The compound 1e is

[0049] In this invention, the mixing temperature of the sodium borohydride solution is -2 to 2°C, preferably 0°C.

[0050] In this invention, the concentration of the sodium hydroxide solution is 8-11 wt%, preferably 9-10 wt%, and more preferably 10 wt%; the amount of sodium hydroxide solution used is preferably 10 drops.

[0051] In this invention, in step (3), the reaction temperature is 50-65°C, preferably 55-65°C, and more preferably 60°C; the reaction time is 2-4 hours, preferably 2.5-3.5 hours, and more preferably 3 hours.

[0052] In this invention, in step (4), the mass-to-volume ratio of compound 1f, sodium acetate and acetic anhydride is 38-45g: 7-10g: 200-230mL, preferably 39-44g: 7.5-9.5g: 205-225mL, and more preferably 40-43g: 8-9g: 210-220mL;

[0053] The compound 1f is

[0054] In this invention, the reaction temperature is 120–140°C, preferably 125–140°C, and more preferably 130–140°C; the reaction time is 2–4 h, preferably 2.5–3.5 h, and more preferably 3 h.

[0055] In this invention, the reaction route is as follows (compound 1 is a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound):

[0056]

[0057] This invention provides the application of 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound in the preparation of antiepileptic drugs.

[0058] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0059] Example 1

[0060] 25.00 g of compound 1a was dissolved in 150 mL of dichloromethane under nitrogen protection and stirred until homogeneous. Then, compound 1b was added dropwise to carry out the reaction. The amount of compound 1b used was 95.08 g. The reaction temperature was 25 °C and the reaction was carried out under stirring until no more bubbles were generated under stirring. The reaction was then stopped. After concentration, 25.00 g of orange-yellow liquid was obtained, which is compound 1c, with a yield of 84.4%.

[0061] 25.00 g of compound 1c and 31.95 g of compound 1d were dissolved in 100 mL of dichloromethane. When the mixture was cooled to 0 °C, 28.12 g of aluminum trichloride was added. The reaction system was then heated to 25 °C and reacted for 2.5 h. The reaction product was then quenched with 300 mL of water and extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to obtain the crude product. The crude product was purified by slurrying with 40 mL of ethanol for 1 hour to obtain 43.70 g of white solid, which was compound 1e, with a yield of 91.9%.

[0062] 43.70 g of compound 1e was dissolved in 150 mL of tetrahydrofuran. The mixture was cooled to 0 °C and 20 mL of sodium borohydride aqueous solution (0.925 g / mL) was added dropwise. Then, 10 drops of 10 wt% sodium hydroxide solution were added. The reaction system was then heated to 60 °C and reacted for 3 h. After cooling to 37 °C, the pH was adjusted to 7 with 1 M hydrochloric acid solution. The tetrahydrofuran was removed by concentration, and the mixture was extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to obtain 42.59 g of orange-yellow semi-solid, which was compound 1f, with a yield of 96.7%.

[0063] 42.59 g of compound 1f and 8.31 g of anhydrous sodium acetate were dissolved in 210 mL of acetic anhydride. The reaction system was heated to 140 °C and reacted for 3 hours. The acetic anhydride was then removed by concentration, and the mixture was quenched with 200 mL of water. The mixture was extracted with ethyl acetate (4 × 200 mL). The organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to obtain the crude product. Finally, the crude product was purified by recrystallization from 70% ethanol to obtain 23.47 g of 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound as a white solid with a yield of 59.4%.

[0064] 1H-NMR (400MHz, CDCl3) δ6.82 (s, 1H), 6.51 (s, 1H), 6.34 (t, J = 2.4Hz, 1H), 3.88 (s, 3H), 3. 82 (s, 3H), 3.81 (s, 3H), 3.13~2.95 (m, 2H), 2.89 (d, J=6.1Hz, 2H), 2.09 (p, J=7.8Hz, 2H).

[0065] 13 C-NMR (100MHz, CDCl3) δ150.7, 148.0, 143.1, 142.5, 119.0, 114.5, 111.4, 98.0, 56.9, 56.6, 56.2, 32.8, 32.6, 18.5.

[0066] Example 2

[0067] 27.00 g of compound 1a was dissolved in 170 mL of dichloromethane under nitrogen protection and stirred until homogeneous. Then, 97 g of compound 1b was added dropwise to carry out the reaction. The reaction temperature was 30 °C and the reaction was carried out under stirring until no more bubbles were generated. The reaction was then concentrated to obtain 26.62 g of orange-yellow liquid, which is compound 1c, with a yield of 83.6%.

[0068] 26.62 g of compound 1c and 33 g of compound 1d were dissolved in 120 mL of dichloromethane. When the mixture was cooled to 0 °C, 30 g of aluminum trichloride was added. The reaction system was then heated to 30 °C and reacted for 2 h. The reaction product was then quenched with 300 mL of water and extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to obtain the crude product. The crude product was purified by slurrying with 40 mL of ethanol for 1 hour to obtain 44.05 g of white solid, which was compound 1e, with a yield of 89.7%.

[0069] 44.05 g of compound 1e was dissolved in 160 mL of tetrahydrofuran. The resulting mixture was cooled to 0 °C, and 22 mL of sodium borohydride aqueous solution (concentration 1.0 g / mL) was added dropwise. Then, 10 drops of 10 wt% sodium hydroxide solution were added dropwise. The reaction system was then heated to 65 °C and reacted for 2 h. After cooling to 37 °C, the pH was adjusted to 8 with 1 M hydrochloric acid solution. The tetrahydrofuran was removed by concentration, and the mixture was extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to obtain 40.10 g of orange-yellow semi-solid, which was compound 1f, with a yield of 90.3%.

[0070] 40.10 g of compound 1f and 10 g of anhydrous sodium acetate were dissolved in 230 mL of acetic anhydride. The reaction system was heated to 140 °C and reacted for 2 hours. The acetic anhydride was then removed by concentration, and the mixture was quenched with 200 mL of water. The mixture was extracted with ethyl acetate (4 × 200 mL). The organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to obtain the crude product. Finally, the crude product was purified by recrystallization from 70% ethanol to obtain 22.59 g of 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound as a white solid with a yield of 59.2%.

[0071] Example 3

[0072] 23.00 g of compound 1a was dissolved in 130 mL of dichloromethane under nitrogen protection and stirred until homogeneous. Then, 93 g of compound 1b was added dropwise to carry out the reaction. The reaction temperature was 20 °C and the reaction was carried out under stirring until no more bubbles were generated. The reaction was then concentrated to obtain 22.87 g of orange-yellow liquid, which is compound 1c, with a yield of 84.0%.

[0073] 22.87 g of compound 1c and 30 g of compound 1d were dissolved in 80 mL of dichloromethane. When the mixture was cooled to 0 °C, 26 g of aluminum trichloride was added. The reaction system was then heated to 20 °C and reacted for 3 h. The reaction product was then quenched with 300 mL of water and extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to obtain the crude product. The crude product was purified by slurrying with 40 mL of ethanol for 1 hour to obtain 40.22 g of white solid, which was compound 1e, with a yield of 90.1%.

[0074] 40.22 g of compound 1e was dissolved in 140 mL of tetrahydrofuran. The mixture was cooled to 0 °C and 18 mL of sodium borohydride aqueous solution (0.8 g / mL) was added dropwise. Then, 10 drops of 10 wt% sodium hydroxide solution were added. The reaction system was then heated to 50 °C and reacted for 4 h. After cooling to 37 °C, the pH was adjusted to 7 with 1 M hydrochloric acid solution. The tetrahydrofuran was removed by concentration, and the mixture was extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, dried with magnesium sulfate, and concentrated to give 38.31 g of orange-yellow semi-solid, which was compound 1f, with a yield of 94.5%.

[0075] 38.31 g of compound 1f and 7 g of anhydrous sodium acetate were dissolved in 200 mL of acetic anhydride. The reaction system was heated to 120 °C and reacted for 4 hours. After concentration to remove acetic anhydride, 200 mL of water was added to quench the reaction, and the mixture was extracted with ethyl acetate (4 × 200 mL). The organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to obtain the crude product. Finally, the crude product was purified by recrystallization from 70% ethanol to obtain 21.10 g of 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound as a white solid with a yield of 59.3%.

[0076] Performance testing

[0077] To verify the protective effect of 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene (compound 1, English name 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene, hereinafter referred to as CTMB) on a pentylenetetrazol-induced epilepsy mouse model and on a lithium-pilocarpine-induced epilepsy rat model.

[0078] CTMB emulsion injection mainly consists of 5%–30% soybean oil, 0.6%–1.8% egg yolk lecithin, 1.0%–2.5% glycerol, appropriate amount of pH adjuster, 80%–95% water, and 0.1%–2% 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene compound. It can be prepared according to the conventional methods for drug-loaded emulsion injections. Unless otherwise specified, CTMB is administered via emulsion injection.

[0079] Test 1

[0080] To verify the protective effect of CTMB on a pentylenetetrazol-induced epilepsy mouse model:

[0081] (1) Reagents and materials

[0082] SPF grade adult male KM mice, weighing 30-35g.

[0083] (2) Grouping of animals

[0084] The mice were randomly divided into three groups: a saline group, a low-dose CTMB group (50 mg / kg, CTMB-L), a medium-dose CTMB group (75 mg / kg, CTMB-M), and a high-dose CTMB group (100 mg / kg, CTMB-H), with 10 mice in each group. All mice were administered the CTMB via intraperitoneal injection. The CTMB in each group was 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene.

[0085] (3) Evaluation methods and results

[0086] After a period of time following administration of the drug, mice in each group were subsequently injected intraperitoneally with 75 mg / kg of pentylenetetrazole, a drug used to induce epilepsy. Clonic and tonic seizures were used as evaluation indicators. The Racine score, as well as the incidence and latency of clonic seizures (Racine score 1–3) and tonic seizures (Racine score 4–5), were observed and recorded within 30 minutes in each group. Racine grading criteria: Grade 0: Normal non-epileptic activity; Grade 1: Wet dog-like shaking or scratching; Grade 2: Head nodding or tail wagging; Grade 3: Unilateral forelimb extension or unilateral limb clonic contractions; Grade 4: Multiple limb clonic or tonic contractions; Grade 5: Falling and generalized tonic-clonic seizures. If no epilepsy occurred within 30 minutes, the latency was recorded as 1800 s. Experimental results are shown in Table 1.

[0087] Table 1. Protective effect of CTMB on a pentylenetetrazol-induced mouse model of epilepsy.

[0088]

[0089]

[0090] Compared with the saline group, * P<0.05, ** P<0.01, *** P<0.001.

[0091] As shown in Table 1, CTMB can significantly prolong the latency of clonic and tonic seizures induced by pentylenetetrazol, reduce the seizure severity and incidence, and its antiepileptic effect is dose-dependent.

[0092] Test 2

[0093] Verify the protective effect of CTMB on a lithium-pilocarpine rat epilepsy model:

[0094] (1) Laboratory animals

[0095] SPF grade adult male SD rats, weighing 200-250g.

[0096] (2) Establishment of a lithium-pilocarpine rat epilepsy model

[0097] Weigh out methylhyoscyamine bromide, lithium chloride, and pilocarpine, and dissolve them in 0.9% physiological saline to the working concentration (lithium chloride solution 100 mg / mL, methylhyoscyamine bromide solution 1 mg / mL, pilocarpine solution 20 mg / mL). Intraperitoneally inject rats with lithium chloride (127 mg / kg), followed by intraperitoneal injection of methylhyoscyamine bromide 1 mg / kg 18–20 h later. Thirty min later, administer an initial intraperitoneal injection of 30 mg / kg pilocarpine, and observe the severity of epileptiform seizures. Thereafter, administer booster doses of 10 mg / kg pilocarpine intraperitoneally every 30 min until grade IV or higher, uninterrupted SE-like seizures occur, with a maximum dose of 60 mg / kg.

[0098] Model selection criteria

[0099] The modeling can be considered successful if experimental animals exhibit grade IV or higher seizures for 1 consecutive hour.

[0100] (3) Animal grouping

[0101] Rats that successfully developed the SE model were randomly divided into three groups: the model group, the CTMB group, and the sodium valproate (VPA) group. Drug administration was performed 1 hour after the SE attack. The model group received a blank emulsion via intraperitoneal injection, the treatment group received 50 mg / kg CTMB emulsion via intraperitoneal injection, and the VPA group received 300 mg / kg sodium valproate via oral gavage.

[0102] (4) Evaluation methods and results

[0103] Post-seizure behavioral assessment following drug intervention:

[0104] Following the Racine grading criteria, the epileptic seizure symptoms of rats in each group after drug intervention were observed, and the seizure severity in the acute phase was recorded. Racine grading criteria: Grade 0: Normal non-epileptic activity; Grade 1: Wet dog-like shaking or scratching; Grade 2: Head nodding or tail wagging; Grade 3: Unilateral forelimb extension or unilateral limb clonic contractions; Grade 4: Multiple limb clonic or tonic seizures; Grade 5: Falling and generalized tonic-clonic seizures; Grade 6: Severe, fatal seizures. The results are shown in Table 2.

[0105] Table 2. Effects of CTMB on the highest seizure severity score in SE model rats.

[0106] Seizure Score 5.44±0.53 4.38±0.52*** <![CDATA[5.00 # ]]>

[0107] Note: For comparison between the CTMB group and the model group, ***P<0.001, t-test; for comparison between the CTMB group and the VPA group, ... # P<0.05, t-test.

[0108] As shown in Table 2, CTMB (50 mg / kg) can significantly reduce the seizure severity in the acute phase of the SE model, and its effect is significantly better than that of the positive control drug sodium valproate (300 mg / kg).

[0109] One-week survival rate observation

[0110] The survival of SE model rats after acute phase drug intervention was observed for two weeks, and the results were as follows: Figure 1 As shown.

[0111] The differences in survival curves among the rat groups were analyzed using the log-rank test. The results showed that CTMB (50 mg / kg) significantly improved the survival rate of SE model rats (P<0.05), and its effect was better than that of the positive control drug sodium valproate (VPA).

[0112] As can be seen from the above examples and performance test results, the present invention provides a 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound, its preparation method and application. The 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound prepared by the present invention can be formulated into a suitable preparation with pharmaceutically acceptable excipients, and can play the following roles: (1) prolonging the latency of clonic and tonic seizures induced by pentylenetetrazol, reducing the seizure severity and incidence, and its anti-epileptic effect is dose-dependent; (2) significantly reducing the seizure severity in the acute phase of the rat epilepsy model induced by lithium-pilocarpine, improving the survival rate of rats, and the effect is better than that of the broad-spectrum anti-epileptic drug sodium valproate.

[0113] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene compound, characterized in that, The 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound has the structure shown in formula (Ⅰ):

2. The method for preparing the 1-(cyclobutylidenemethyl)-2,4,5-trimethoxybenzene compound according to claim 1, characterized in that, Includes the following steps: (1) Compound 1a was mixed with dichloromethane, and then compound 1b was added to react and compound 1c was obtained. (2) Compound 1c, compound 1d, and aluminum trichloride were mixed with dichloromethane in sequence and reacted to obtain compound 1e; (3) Compound 1e, sodium borohydride solution, and sodium hydroxide solution were mixed with tetrahydrofuran in sequence and reacted to obtain compound 1f; (4) Compound 1f, sodium acetate and acetic anhydride were mixed and reacted to obtain 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound; The compound 1a is The compound 1b is The compound 1d is 3. The preparation method according to claim 2, characterized in that, In step (1), the mass-to-volume ratio of compound 1a, dichloromethane, and compound 1b is 23–27 g: 130–170 mL: 93–97 g.

4. The preparation method according to claim 2 or 3, characterized in that, In step (1), the reaction temperature is 20-30°C, and the reaction is carried out under stirring conditions until no more bubbles are generated under stirring conditions and the reaction ends.

5. The preparation method according to claim 4, characterized in that, In step (2), the mass-to-volume ratio of compound 1c, compound 1d, aluminum trichloride and dichloromethane is 22-27g: 30-33g: 26-30g: 80-120mL; The compound 1c is 6. The preparation method according to claim 2, 3, or 5, characterized in that, In step (2), the reaction temperature is 20-30°C and the reaction time is 2-3 hours.

7. The preparation method according to claim 6, characterized in that, In step (3), the mass-to-volume ratio of compound 1e, sodium borohydride solution, and tetrahydrofuran is 40-45 g: 18-22 mL: 140-160 mL, wherein the concentration of sodium borohydride solution is 0.8-1.0 g / mL. The compound 1e is 8. The preparation method according to claim 5 or 7, characterized in that, In step (3), the reaction temperature is 50-65℃ and the reaction time is 2-4h.

9. The preparation method according to claim 8, characterized in that, In step (4), the mass-to-volume ratio of compound 1f, sodium acetate and acetic anhydride is 38-45g: 7-10g: 200-230mL; The compound 1f is The reaction temperature is 120–140°C, and the reaction time is 2–4 hours.

10. The use of the 1-(cyclobutyrylmethyl)-2,4,5-trimethoxybenzene compound of claim 1 in the preparation of an antiepileptic drug.

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