CN114652720A

CN114652720A - Application of epilupine and derivatives thereof in preparation of medicines for treating depression

Info

Publication number: CN114652720A
Application number: CN202210161755.4A
Authority: CN
Inventors: 杜静; 吉腾飞; 阚伟京; 王刚
Original assignee: Institute of Materia Medica of CAMS; Beijing Anding Hospital
Current assignee: Institute of Materia Medica of CAMS; Beijing Anding Hospital
Priority date: 2022-02-22
Filing date: 2022-02-22
Publication date: 2022-06-24
Anticipated expiration: 2042-02-22
Also published as: CN114652720B

Abstract

The invention provides application of epilupine (1-epilupine) and derivatives thereof in preparing a medicament for treating depression. The application dosage is 0.01-30mg of epilupine or derivatives thereof per kilogram of body weight, and the antidepressant effect can be produced within 60 minutes after administration. After 5 days of administration, both low and high doses produced significant antidepressant effects. The research result of the invention shows that in the tail suspension experiment of the mammal mouse, the epilupine or the derivative thereof has the effect of quickly resisting depression and is more effective than the conventional antidepressant drug imipramine. The invention adopts forced swimming experiment and mouse tail suspension experiment as drug screening experiment, and simultaneously carries out mouse Open field test (Open field test) to test the autonomous activity of the mouse, thereby avoiding the interference of central stimulant. The antidepressant drug has the advantages of obvious antidepressant effect, quick response, small dosage, small side effect and the like, and is expected to become a new antidepressant drug with high speed, high efficiency and a new mechanism.

Description

Application of epilupine and derivatives thereof in preparation of medicines for treating depression

Technical Field

The invention belongs to the technical field of medicines, and relates to application of epilupine (1-epilupine) and derivatives thereof in preparation of medicines for treating depression and prepared antidepressant medicines.

Background

Major Depressive Disorder (MDD) is a serious, recurrent disabling psychotic disorder that accounts for approximately 3 billion depression worldwide. The incidence of depression in china is 4.2%, with about 5600 ten thousand patients suffering depression. Depression is the second leading cause of life loss (years) due to disability, and has become a major public health problem that urgently needs to be solved. Depression is clinically manifested as depressed mood, hyposis, and even pessimistic and boredom, with suicide attempts or behaviors. The current first-line clinical antidepressants mainly comprise selective 5-hydroxytryptamine reuptake inhibitors, 5-hydroxytryptamine and norepinephrine reuptake inhibitors and the like, but the drugs have slow effect, narrow action spectrum and easy relapse after drug withdrawal. Despite the use of multiple antidepressant medications, about 30% of patients become refractory Depression (TRD). Patients with this type of refractory depression present a higher risk of social burden and risk of suicide. Therefore, the search for new antidepressant drugs with good compliance, few side effects and novel pharmacological mechanisms has great clinical requirements, and is also a hot spot of international research at present.

Epilonine (1-Epilupinine), also known as Epilupinine, is isolated from lupine (Lupinus polyphyllus) belonging to the genus Lupinus of the family Leguminosae. Epilotoinine is a quinolizidine alkaloid with molecular formula C₁₀H₁₉NO, molecular weight 169.26, melting point 77-78 deg.C, and dissolving in chloroform, methanol, and ethanol. Can be used for preparing chemical pesticide.

The subacute toxicity test proves that the epilupine has no obvious influence on the functions of the bone marrow, the liver, the kidney and the like of the rat. At present, reports of using epilupine as an active ingredient for preventing or treating depression are not seen.

Disclosure of Invention

The invention aims to provide an application of epilupine and derivatives thereof in preparing a medicament for treating depression, and the medicament has the advantages of remarkable anti-depression effect, quick response, small dosage, small side effect and the like.

The invention provides an application of epilupine or derivatives thereof in preparing a medicament for treating depression,

further, the derivatives include: organic/inorganic acid salt type products of lupine, and esterification products or etherification products of 11-position hydroxyl group.

Further, the dosage for application is 0.01-30mg of epilupine or its derivative per kg of body weight.

The invention also provides an anti-depression drug, which comprises the following components: epilupine or its derivatives, and pharmaceutically acceptable auxiliary components.

The medicinal components also comprise medicinal components which have positive effects on treating depression after being simultaneously applied.

Further, the pharmaceutical ingredients which have positive effects on treating depression after simultaneous administration are as follows: monoamine oxidase inhibitor MAOI, tricyclic antidepressant TCA, selective serotonin reuptake inhibitor SSRI, serotonin and norepinephrine reuptake inhibitor SNRI, noradrenergic and specific 5-HT receptor antagonist NASSA, 5-HT antagonist and reuptake inhibitor, or melatonin and 5-HT2C receptor antagonist.

Further, tricyclic antidepressants (TCA) including amitriptyline, imipramine, chlorpromazine, doxepin, clomipramine and the like; such Selective Serotonin Reuptake Inhibitors (SSRIs) are for example: fluoxetine (BAUTOU), paroxetine (Cyclet), fluvoxamine (Radil), sertraline (Sertral ine, levofloxacin), citalopram (citalopram, Simplica), escitalopram (escitalopram), etc.; the Serotonin and Norepinephrine Reuptake Inhibitors (SNRI), e.g., venlafaxine, duloxetine, etc.; the noradrenergic and specific 5-HT receptor antagonists (NASSA), e.g., mirtazapine, etc.; the 5-HT antagonist and reuptake inhibitor: such as nefazodone and trazodone; the melatonin and 5-HT2C receptor antagonists: such as agomelatine.

Further, the pharmaceutical ingredients which have a positive effect on the treatment of depression after simultaneous administration are herbs such as: john's wort, and tianeptine, Vortioxetine (also known as Vortioxetine), Bupropion (bupapion), and the like.

Further, the pharmaceutical ingredients which have positive effects on treating depression after simultaneous administration are as follows: ketamines, including S-ketamine or ketamine nasal feeds.

Further, the effective dose of the medicine is 0.01-30mg of epilupine and derivatives thereof per kilogram of body weight.

The above medicinal preparation is in the form of powder, granule, tablet, capsule, pill, solution, suspension or injection.

The epilupine is white powder, HRESI-MS M/z [ M + H ]]⁺170.1539, it is suggested that its molecular formula is C₁₀H₁₉NO。

¹H NMR(400MHz,CDCl₃)δ3.73-3.46(m,1H),2.86-2.68(m,1H),2.01(ddd,J＝14.6,11.4,2.9Hz,1H),1.92-1.79(m,1H),1.79–1.63(m,1H),1.59(t,J＝3.4Hz,1H),1.51-1.35(m,OH),1.30-1.09(m,1H).

¹³C NMR(100MHz,CDCl₃)δ64.67,64.31,56.92,56.64,43.96,29.78,28.23,25.60,25.04,24.58。

The medicament can be administered in unit dosage form, and the administration dosage form can be liquid dosage form or solid dosage form. For example, the liquid dosage form can be true solution, colloid, microparticle, emulsion, or suspension. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, powder, solution, suspension, emulsion, granule, suppository, lyophilized powder for injection, etc.

The administration route can be intestinal or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosal, cutaneous, peritoneal or rectal.

The route of administration of the drug of the present invention may be administration by injection. The injection includes intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, acupoint injection, etc.

The medicine of the present invention may be prepared into common preparation, slow released preparation, controlled releasing preparation, targeting preparation and various particle medicine feeding system.

In order to prepare the unit dosage form into tablets, various carriers well known in the art can be widely used. In the vector: diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate, and the like; wetting agents and binders such as water, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrating agents such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecylsulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors such as sucrose, glyceryl tristearate, cacao butter, hydrogenated oil and the like; absorption accelerators such as quaternary ammonium salts, sodium lauryl sulfate and the like; lubricants, for example, talc, silica, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, and the like. The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.

For making the administration units into pills, a wide variety of carriers well known in the art can be used. In the vector: for example, diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oils, polyvinylpyrrolidone, Gelucire, kaolin, talc, and the like; binders such as acacia, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste or batter, etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecylsulfate, methylcellulose, ethylcellulose, etc.

For making the administration unit into a suppository, various carriers well known in the art can be widely used. In the vector: such as polyethylene glycol, lecithin, cocoa butter, higher alcohols, higher alcohol enzymes, gelatin, semisynthetic glyceroses, and the like.

To encapsulate the administration units, the active ingredient is mixed with the various carriers described above, and the mixture thus obtained is placed in hard gelatin capsules or soft gelatin capsules. Or making into microcapsule, suspending in aqueous medium to form suspension, or making into hard capsule or injection.

For example, the composition of the present invention is formulated into an injectable preparation, such as a solution, a suspension solution, an emulsion, a lyophilized powder, which may be aqueous or non-aqueous, and may contain one or more pharmaceutically acceptable carriers, diluents, binders, lubricants, preservatives, surfactants or dispersants. For example, the diluent may be selected from water, ethanol, polyethylene glycol, 1, 3-propanediol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitol fatty acid enzyme, etc. In addition, for the preparation of isotonic injection, an appropriate amount of sodium chloride, glucose or glycerol may be added to the preparation for injection, and in addition, a conventional cosolvent, a buffer, a pH adjuster, and the like may be added. These adjuvants are conventional in the art.

In addition, if desired, colorants, preservatives, flavors, flavorings, sweeteners, or other materials may also be added to the pharmaceutical preparation.

The dose of the pharmaceutical composition of the present invention to be administered depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, body weight, character and individual response of the patient or animal, the administration route, the number of administrations, etc., and thus the therapeutic dose of the present invention can be widely varied. Generally, the dosage of the compounds of the present invention used is well known to those skilled in the art. The actual effective amount of the drug contained in the final preparation of the medicinal composition can be properly adjusted according to the requirement of the effective amount of the drug so as to achieve the treatment of obesity, abnormal glucose tolerance, hyperlipidemia, diabetes, complications thereof and other diseases; and the use of chronic inflammation, insulin resistance, cerebrovascular disease and atherosclerosis in relation to the above mentioned diseases.

The invention provides an anti-depression medicament, which comprises two stages of prevention and treatment. The dosage of the anti-depression drug in the prevention stage is as follows: 0.001-3mg per kg body weight per day; the dosage of the anti-depression drug in the treatment period is as follows: providing 0.01-30mg of epilupine (1-epilupine) and derivatives thereof per kg of body weight per day. The administration mode of the anti-depression drug is oral administration, nasal spray, instillation or injection; the subject to be administered is a mammal, including a human.

The anti-depression drug provided by the invention is used for treating depression, and the dosage of the epilupine (1-epilupine) and the derivative thereof is 0.01-30mg per kilogram of body weight; the antidepressant effect can be achieved within 60 minutes after administration. After 5 days of administration, both low and high doses produced significant antidepressant effects. The medicine has the advantages of remarkable antidepressant effect, quick action, small dosage, small side effect and the like. In forced swimming experiments of mammal mice, the epilupine (1-epilupine) and the derivatives thereof have more remarkable antidepressant effect than the conventional antidepressant drug imipramine.

Compared with the prior art, the invention has the beneficial technical effects that:

1. the invention provides application of epilupine (1-epilupine) or derivatives thereof in preparing anti-depression drugs. The research result of the invention shows that in the tail suspension experiment of a mammal mouse, the epilupine (1-epilupine) or the derivative thereof has the effect of quickly resisting depression and is more effective than the conventional antidepressant drug imipramine.

2. The invention adopts forced swimming experiment and mouse tail suspension experiment as drug screening experiment. Forced swimming experiments and mouse tail suspension experiments are two common animal behavior despair depression model experiments, and can better ensure the reliability of screening results. Forced swimming experiments in mice have been used for screening of many antidepressants. Moreover, most of the antidepressants with clinical treatment effects are also proved to be effective in reducing immobility time in forced swimming experiments.

By immobile, it is meant that "the animal stops struggling in water, or is in a floating state, only exposing nostrils to keep breathing, and only having small limb movements to keep the head floating on the water". Drugs to be screened were administered prior to the assay. Animals cannot escape from the harsh environment due to downstream swimming in a forced state, resulting in despair behavior of the animals. The model method is simple, convenient and reliable, and is widely used for screening and evaluating the antidepressant. The tail suspension experiment of the mouse is that the mouse does not struggle under the tail suspension state and presents a special quiet immobility state, and the antidepressant can obviously shorten the duration time of the immobility state. During the test, the tail of the mouse is fixed and suspended upside down. The tail of the mouse is not twisted and folded. And recording the immobility time. The immobility indexes are as follows: animal limbs and trunk struggle without twisting. The tail suspension experiment is sensitive to various antidepressants, and avoids the interference of temperature and animal movement dysfunction in the swimming experiment, so that the results of the forced swimming experiment can be effectively verified and supplemented when some mice are used for screening the antidepressants.

3. Shows stronger antidepressant effect than imipramine. In a tail suspension experiment of a mouse animal model, the epilupine (1-epilupine) and derivative thereof in a high-dose group mouse (10mg/kg) have the effect of relieving depression symptoms 60 minutes after injection, namely the activity is more frequent than that of a blank control group, and the immobility time is obviously shortened. Epilupine (1-epilupine) and its derivatives started to significantly shorten the immobility time of tail suspension 5 days after injection. Epilonine (1-epilupine) and its derivatives show a greater antidepressant effect than imipramine at doses (10mg/kg) lower than that of imipramine (15 mg/kg).

4. The invention also carries out an Open field test (Open field test) of the mouse to check the autonomous activity of the mouse and avoid the interference of central stimulant. Since the shortening of the immobility time of animals in classical animal models of depression may be due to the central excitatory action of the drug, the present invention also simultaneously performs mouse open-field experiments. The results show that high concentration of lupine (1-epilupine) and derivatives thereof have no significant effect on the autonomic activity of mice compared to the control group, so that the possibility of mania caused by the stimulant can be excluded, and that the lupine (1-epilupine) and derivatives thereof indeed have significant antidepressant effect.

5. The antidepressant drug taking the epilupine (1-epilupine) or the derivative thereof as the active ingredient is expected to become a new antidepressant drug with high speed, high efficiency and a new mechanism.

Drawings

FIG. 1 shows structural formula of epilupine (1-epilupine);

FIG. 2. epilupine (1-epilupine)¹H NMR；

FIG. 3 preparation of epilupine (1-epilupine)¹³C NMR；

FIG. 4 MS of epilupine (1-epilupine);

FIG. 5. Effect of epilupine (1-epilupine) injection on mice tail suspension test one hour later;

ordinate: tail suspension experiment mice immobility time(s), abscissa: from left to right, there were a DMSO control group, an epilupine (1-epilupine) low dose group (ELP-L, 5mg/kg), an epilupine (1-epilupine) high dose group (ELP-H,10 mg/kg), a saline control group, and a imipramine positive control group (15 mg/kg).

FIG. 6 Effect of epilupine (1-epilupine) injection on the swimming test of mice 5 days after injection

Ordinate: forced swimming test mice immobility time(s), abscissa: from left to right in sequence: DMSO control group, epi-lupine (1-epilupine) low dose group (ELP-L, 5mg/kg), epi-lupine (1-epilupine) high dose group (ELP-H,10 mg/kg), saline control group, imipramine positive control group (IMI, 15 mg/kg).

Detailed Description

The present invention is described in detail below by way of specific examples, it being understood that the following examples are intended only by way of illustration and description and are not intended to limit the scope of the present invention in any way. In the following embodiments, the biochemical reagents not specifically described are all conventional reagents in the art, and may be prepared according to conventional methods in the art or commercially available, and may be of laboratory pure grade.

Epilupine (1-epilupine), CAS No.: 486-71-5, made by the laboratory.

Example 1:

10 kg of lupin (Lupinus micropranthus) seeds are crushed, heated and refluxed by 95 percent ethanol for 3 times, 50 liters of the extract is carried out each time, the extraction time is two hours each time, the extract is decompressed and concentrated to obtain 2.5 kg of extract, the 95 percent ethanol extract is suspended in 6 liters of distilled water, the pH value of the aqueous solution is adjusted to 1-3 by 2mol/L dilute hydrochloric acid solution, and the aqueous solution is extracted by chloroform for 3 times, 6 liters each time; the aqueous acid solution was adjusted to pH L0-11 with 2mol/L NaOH solution and extracted 3 times with 6 liters each time with chloroform. Chloroform was recovered to obtain total alkaloid 80.0 g. The total alkaloids are subjected to silica gel column chromatography for coarse separation, and gradient elution with chloroform-methanol system (10:0-0:10) to obtain 6 fractions (Frs.A-F). Wherein Fr.B (9.6g) is separated and purified by silica gel column chromatography and Sephadex LH-20 to obtain 2g of compound epilupine (1-epilupine). Melting point of 77-78 deg.C, and dissolving in chloroform, methanol, and ethanol; HR-ESI-MS M/z [ M + H ]]⁺170.1539, it is suggested that its molecular formula is C₁₀H₁₉NO。

Example 2:

taking 150g of epilupine and a proper amount of auxiliary materials, sieving, uniformly mixing, and encapsulating to prepare 1000 epilupine capsules. And obtaining the content of epilupine according to quantitative analysis, and taking the content as the basis of the dosage.

Example 3:

diluting epilupine 150g with water, and flavoring with stevioside to obtain 10% liquid preparation, and making into epilupine oral liquid. And obtaining the content of epilupine according to quantitative analysis, and taking the content as the basis of the dosage.

Example 4:

dissolving 100 g of epilupine in 500 ml of absolute ethyl alcohol, dropwise adding a 3% dilute hydrochloric acid solution until the pH value is 6, standing to separate out epilupine hydrochloride crystals, filtering, and drying to obtain the epilupine hydrochloride; adding water for injection, fine filtering, bottling, and sterilizing to obtain injection.

Example 5:

dissolving 200 g of epilupine in 1000 ml of absolute ethanol, dropwise adding a 5% sulfuric acid ethanol solution until the pH value is 6, standing, separating out crystals, filtering and drying to obtain epilupine hydrochloride; dissolving in sterile water for injection, adding excipient, stirring to dissolve, filtering with sterile filter funnel, sterile fine filtering, packaging into 2 ampoules, freeze drying at low temperature, sterilizing, and sealing to obtain powder for injection.

Example 6:

adding excipient into the epilupine according to the weight ratio of the epilupine to the excipient of 9:1, and preparing into powder.

Example 7:

adding excipient according to the weight ratio of the epilupine to the excipient of 1:5-1:10, granulating and tabletting.

Example 8:

dissolving 50g of epilupine in 300 ml of absolute ethanol, dropwise adding a 3% phosphoric acid ethanol solution until the pH value is 6, standing, separating out crystals, filtering and drying to obtain epilupine phosphate; making into oral liquid by conventional method.

Example 9:

adding excipient into the epilupine according to the weight ratio of the epilupine to the excipient of 5:1, and making into capsule, granule or granule.

Example 10:

adding excipient into the epilupine according to the weight ratio of the epilupine to the excipient of 3:1, and making into capsule, granule or granule.

Example 11:

46.6 g of epilupine, 600 g of starch, 200 g of lactose, 3 g of menthol and 152 g of sodium carboxymethyl starch are added to prepare the buccal tablet which is used as a functional food.

Test example 1 Effect of Epilocamine (1-epilupine) on tail suspension test in animal depression model

Experimental animals: c57BL/6 mice, male, 30-40 grams in weight, supplied by sbefu (beijing) biotechnology limited, license number: SCXK (Jing) 2019-. The animals are raised in cages with a light and shade period of 12h/12h, the room temperature is 20-22 ℃, the water is free, and the feed is provided by the center of the large protein laboratory animals. Experimental drugs: epilonine (1-epilupine) is extracted from institute of medicine of Chinese academy of medical sciences, and its chemical purity and structure analysis are shown in FIGS. 1, 2, 3, and 4. Imipramine dihydrochloride (Imipramine hydrochloride) is a product of Sigma, the product number is I7379, and the batch number is O56K 1380. Experimental equipment: a cross bar; rubberized fabric; a camera (a mobile phone); JUNSO multifunctional timer. The experimental steps are as follows: male C57BL/6 mice were acclimatized for one week and divided into 5 groups of 6-8 mice each. The control group comprises a dimethyl sulfoxide DMSO control group (DMSO dissolved in normal saline), an epilupine (1-epilupine) low dose group (ELP-L,5mg/kg dissolved in DMSO), an epilupine (1-epilupine) high dose group (ELP-H,10mg/kg dissolved in DMSO), a traditional antidepressant drug imipramine positive control group (15mg/kg imipramine dissolved in normal saline) and a normal saline control group. The medicine is administrated by intraperitoneal injection at 10 am according to 0.1ml/10g body weight, and after 60 minutes of injection, tail suspension experiments are carried out on 6-8 mice in each group; in the tail suspension experiment of the mouse, the tail of the mouse is stuck on a horizontal cross bar at a position 2 cm away from the tail tip by using an adhesive tape, so that the animal is in an inverted hanging state, and the head of the animal is 15 cm away from a table top. The observation time was 6 minutes and the cumulative immobility time was recorded over 4 minutes. The immobility indexes are as follows: neither the animal's limbs nor body struggles. The statistical method comprises the following steps: the results are expressed as mean ± SE and three samples were tested by ANOVA. The average comparison of the two samples (imipramine and saline) was performed using the t-test. The mice in the high dose group of epilupine (1-epilupine) had initial relief of depression symptoms at 60 minutes after injection, i.e. struggling activity was more and more frequent than in the control group, significantly reducing immobility time. The activity of the imipramine positive control group and the physiological saline control group is not enhanced to be obvious within 60 minutes after injection. Results were observed 60 minutes after injection and: c57BL/6 male mice showed significant antidepressant effects in the mice tail suspension experiments 60 minutes after intraperitoneal injection of different doses of epilupine (1-epilupine) (10mg/kg) in the high dose group (animals per group N-6-10, ANOVA test p < 0.05, p < 0.01). As shown in figure 5, compared with DMSO control group, the immobility time of mice in high dose group of epilupine (1-epilupine) is reduced from 191.70 + -4.21 s to 164.6 + -6.39 s (P < 0.05) and 160.2 + -6.39 s, and the result shows that the epilupine (1-epilupine) can take effect in a short time, has obvious effect and can resist depression despair symptoms caused by forced tail suspension of the mice.

Test example 2 Effect of Epilocamine (1-epilupine) on forced swimming test in animal depression model

Experimental animals: c57BL/6 mice, male, offered by sbefu (beijing) biotechnology limited, license number: SCXK (Jing) 2019-. The animals are raised in cages one week before the experiment, the light and shade period is 12h/12h, the room temperature is 20-22 ℃, the water is free, and the feed is provided by the center of the Huada protein experimental animals. Experimental drugs: epilonine (1-epilupine) was provided by the institute of pharmacy, national academy of medicine. Imipramine is a product from Sigma, cat # I7379, lot # O56K 1380. An experimental instrument: glass cylinders (height 40cm, diameter 14 cm); a mouse open box (the length and the width are 50cm, the height is 40cm, and the bottom is 16 equal parts); a thermometer; JUNSO multifunctional timer.

The experimental steps are as follows: drug preparation same tail suspension experiment. Animal drug injection: c57BL/6 mice, age 7 weeks, weigh 30-40 grams, acclimate for one week and begin the experiment. Animals were randomly grouped into 5 groups of 8-10 animals each. Intraperitoneal injections were started at 10 am each day. The injection amount is 0.3 ml/30 g, the injection is performed intraperitoneally once a day, and a forced swimming experiment is started five days after the injection. Forced swimming experiment: each group of C57BL/6 mice was placed individually vertically into a plexiglas cylinder (40cm height by 14cm diameter) at a water depth of 25cm and a water temperature of 21-23 ℃. The administration group and the control group were recorded for 6 minutes, and the cumulative immobility time of each group of mice in the next 4 minutes was compared. And (3) judging the motionless time: the mouse floats on the water surface, does not try to climb out of the cylinder, and only does some action which is necessary to keep the head of the mouse on the water surface.

The statistical analysis method comprises the following steps: the results are expressed as means ± SE and the comparison of the mean of the two samples is performed by t-test. As a result: five days after intraperitoneal injection of different doses of epilupine (1-epilupine) in C57BL/6 male mice, a very significant antidepressant effect was shown in the forced swimming test of the mice. This antidepressant effect is more pronounced than that of the traditional antidepressant drug imipramine (N-8-10, p < 0.05, p < 0.01 for t test in each group of animals). As shown in FIG. 6, the immobility time of both the high dose and low dose groups of mice with epilupine (1-epilupine) was significantly reduced within 4 minutes of forced swimming compared to the DMSO control group, and the immobility time of the low dose and high dose groups of mice with epilupine (1-epilupine) was reduced from 58.13 + -14.00 s to 22.50 + -5.20 s (p < 0.01) and 23.63 + -4.37 s (p < 0.01) to 61.29% and 59.35% respectively compared to the 1% DMSO control group. The antidepressant effect of epilupine (1-epilupine) is greater than that of the traditional antidepressant drug imipramine (reduction of 24.32%).

Experimental example 3. mouse open field experiment:

since the reduction in immobility time of animals in classical animal models of depression may be due to the central excitatory action of drugs, the present invention performs a mouse open field experiment to examine the central excitatory action of epilupine (1-epilupine). The mice were placed in the middle of the open box and video recorded for 6 minutes. The experimental results show that the total distance of 5 min after the mice move in the open box is (15.64 ± 7.50m) and (15.26 ± 3.64m) after the high dose and low dose treatment of epilupine (1-epilupine), respectively, compared with the control group (16.62 ± 4.32m), without significant changes (Anova, p > 0.05, N ═ 21), so that the possibility of inducing mania can be excluded, and thus, the epilupine (1-epilupine) indeed has a significant antidepressant effect.

Finally, the above embodiments are only intended to illustrate the technical solutions of the present invention and not to limit the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions, and all of them should be covered by the claims of the present invention.

Claims

1. The application of the epilupine or the derivative thereof in the preparation of the medicine for treating the depression,

2. the use according to claim 1, wherein the derivative comprises: organic/inorganic acid salt type products of epilupine, and esterification products or etherification products of the 11-position hydroxyl group.

3. Use according to claim 1, wherein said use is carried out in a dose of 0.01-30mg of epilupine or derivative thereof per kg of body weight.

4. An anti-depression drug, wherein the drug comprises the following components: epilupine or its derivatives, and pharmaceutically acceptable auxiliary components.

5. The antidepressant as in claim 4, wherein said pharmaceutical components further comprise pharmaceutical components having a positive effect on the treatment of depression after the simultaneous administration.

6. The antidepressant as claimed in claim 5, characterized in that said pharmaceutical ingredients with a positive effect on the treatment of depression after the simultaneous administration comprise: monoamine oxidase inhibitor MAOI, tricyclic antidepressant TCA, selective serotonin reuptake inhibitor SSRI, serotonin and noradrenaline reuptake inhibitor SNRI, noradrenergic and specific 5-HT receptor antagonist NASSA, 5-HT antagonist and reuptake inhibitor or melatonin and 5-HT2C receptor antagonist.

7. An antidepressant drug according to claim 5, characterized in that said pharmaceutical ingredients having a positive effect on the treatment of depression after the simultaneous administration comprise herbs which are St.

8. The antidepressant drug according to claim 5, characterized in that said pharmaceutical composition having a positive effect on the treatment of depression after the simultaneous administration comprises ketamine, which is S-ketamine or ketamine nasal feed.

9. The antidepressant as claimed in claim 4, 5, 6, 7, 8 or 9, characterized in that said pharmaceutical form is a powder, granules, tablets, capsules, pills, solutions, suspensions or injections.