CN110386858B - Platycodon grandiflorum neoalcohol, and extraction method and medical application thereof - Google Patents

Abstract

The invention relates to platycodon grandiflorum new alcohol, an extraction method and medical application thereof, and belongs to the field of medicines. Taking dry platycodon grandiflorum leaves, crushing, adding 70-90% ethanol in an amount which is 3-5 times that of the crushed platycodon grandiflorum leaves, performing reflux extraction for 3-5 times, performing reflux extraction for 2-4 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, performing 200-mesh 400-mesh silica gel column chromatography on the ethyl acetate extract repeatedly, wherein an eluent is petroleum ether: ethyl acetate (80-1): (1-20); after preparative HPLC, the eluent was methanol: water (10-1): (1-10), and keeping the compound with the retention time of 30-60 min to obtain the platycodon grandiflorum new alcohol. The invention is used for preparing the antidepressant medicament and has the characteristic of remarkable curative effect. The dosage of the effective substance may vary depending on the administration mode, the age and weight of the patient, the severity of the condition, and other similar factors.

Description

Platycodon grandiflorum neoalcohol, and extraction method and medical application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to platycodon grandiflorum neoalcohol, an extraction method thereof and application thereof in preparation of antidepressant medicines.

Background

Platycodon grandiflorum (Jacq.) a.dc.) is a perennial plant of the genus Platycodon (Platycodon) of the family platycodonaceae (Campanulaceae). Is distributed in northeast and northeast China, Japan, Korea, Russia, etc. The ecological environment adaptive growth capability is very strong, the ecological environment adaptive growth capability is mainly achieved in shrubs and sunny herbaceous communities, and the ecological environment adaptive growth capability is rarely achieved in forests with the altitude of less than 2000 m. Platycodon grandiflorum was recorded in Shen nong Ben Cao Jing: "mainly treating chest and hypochondriac pain such as knife thorn, abdominal fullness and intestine sound, palpitation and fright, with pungent and mild taste, leading to lung meridian, and having the efficacy of eliminating phlegm, ventilating lung, relieving sore throat, etc., is a commonly used Chinese medicine.

The platycodon grandiflorum is rich in chemical components, including saponins, flavonoids, sterols, polyacetylenes, phenols, volatile oils, fatty acids, amino acids and the like; modern pharmacological studies show that platycodon grandiflorum has the functions of eliminating phlegm, relieving cough, resisting inflammation, resisting oxidation and tumors, protecting cardiovascular systems, promoting lipid metabolism and the like.

Much research on platycodon grandiflorum has focused on roots, while less research on aerial parts such as stems, leaves and seeds has been conducted.

Disclosure of Invention

The invention provides platycodon grandiflorum new alcohol, an extraction method and medical application thereof.

The invention adopts the technical scheme that platycodon grandiflorum neoalcohol with the following formula:

the molecular formula is as follows: c₁₄H₂₂O₅Molecular weight: 270.1476

Chemical name: 2,3,9,13,14-pentahydroxy-4,6-tetradecadiyne

Brown oil, and is easily soluble in methanol, ethanol, and pyridine. TLC detection, 10% H₂SO₄The ethanol solution is a color developing agent, and the color is blue when the mixture is heated at 105 ℃.

The extraction method of the platycodon grandiflorum neoalcohol comprises the following steps:

taking dry platycodon grandiflorum leaves, crushing, adding 70-90% ethanol in an amount which is 3-5 times that of the crushed platycodon grandiflorum leaves, performing reflux extraction for 3-5 times, performing reflux extraction for 2-4 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, performing 200-mesh 400-mesh silica gel column chromatography on the ethyl acetate extract repeatedly, wherein an eluent is petroleum ether: ethyl acetate (80-1): (1-20); after preparative HPLC, the eluent was methanol: water (10-1): (1-10), and keeping the compound with the retention time of 30-60 min to obtain the platycodon grandiflorum new alcohol.

Preferably:

taking dried platycodon grandiflorum leaves, crushing, adding 5 times of 80% ethanol for reflux extraction for 4 times, extracting for 3 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, subjecting an ethyl acetate extract to repeated 200-mesh silica gel column chromatography, wherein an eluent is petroleum ether: ethyl acetate 1: 1; after preparative HPLC, the eluent was methanol: and (3) keeping the compound with the retention time of 45min when the water is 10:3 to obtain the platycodon grandiflorum neoalcohol.

The invention relates to application of platycodon grandiflorum neoalcohol in preparation of antidepressant drugs.

When the invention is used for preparing the auxiliary medicine for treating depression, the oral administration or the parenteral administration is safe, and in the oral case, the auxiliary medicine can be administered in any conventional form, such as powder, granules, tablets, capsules, pills, solutions, suspensions, syrups, buccal tablets, sublingual tablets and the like: when the drug is administered parenterally, it may take any conventional form, such as injections: such as intravenous injection, ointment, suppository, transdermal administration, inhalant, etc.

The medicament for treating depression of the present invention is composed of an active ingredient monomer or an active ingredient together with a solid or liquid excipient, the solid or liquid excipient used herein is well known in the art, and the powder is an internal powder having, as specific examples, lactose, starch, dextrin, calcium carbonate, synthetic or natural aluminum sulfate, magnesium oxide, magnesium stearate, sodium bicarbonate, dried yeast, etc.; the excipient of the solution agent comprises water, glycerol, 1, 2-propylene glycol, simple syrup, ethanol, ethylene glycol, polyethylene glycol, sorbitol, etc.; the excipient of ointment can be fatty oil, hydrophobic agent or hydrophilic agent composed of aqueous lanolin, vaseline, glycerol, adeps Apis, lignan, liquid petrolatum, resin, and higher wax.

The invention has the beneficial effect that the new compound platycodon grandiflorum new alcohol. Can be used for preparing antidepressant drugs and has the characteristic of remarkable curative effect. The dosage of the effective substance may vary depending on the administration mode, the age and weight of the patient, the severity of the condition, and other similar factors.

Drawings

FIG. 1 is the main HMBC related signal of the compound of the present invention;

FIG. 2 is a HR-ESI-MS spectrum of a compound of the present invention;

FIG. 3 is a drawing of a compound of the present invention¹H NMR spectrum;

FIG. 4 is a DEPTQSP spectrum of a compound of the invention;

FIG. 5 is a HMQC spectrum of a compound of the invention;

FIG. 6 is an HMBC spectrum of a compound of the present invention;

FIG. 7 is a plot of sugar water preference rate of the present invention, where P represents comparison to model groups<0.05; comparison of representation with model group P<0.01；^##Comparison of representatives with the control group P<0.01；

FIG. 8 is a graph showing the change in body weight of a mouse according to the present invention;

figure 9 is a graph of the effect of food intake in mice of the invention, where represents P <0.01 compared to the model group;

figure 10A is a horizontal score of open field movement of mice of the invention, where represents P <0.01 compared to the model group;

figure 10B is a vertical score of the open field movement of mice of the invention, where represents P <0.01 compared to the model group;

FIG. 11 is the immobility time of the forced swim test of mice of the invention, where P <0.01 is represented compared to the model group;

figure 12 is the immobility time of the mouse tail suspension experiment of the invention, wherein represents P <0.01 compared to the model group;

figure 13A is the immobility time of the prazosin pretreated mice forced swim test of the invention, where P <0.01 compared to the model group is represented;

fig. 13B is the immobility time of forced swim experiments for haloperidol pretreated mice of the present invention, wherein x represents P <0.01 compared to model group;

figure 13C is the immobility time of alkali pretreated bicuculline mice forced swim experiments of the invention, represented by P <0.01 compared to model group;

fig. 13D is the immobility time of NMDA pre-treated mice forced swim experiments of the invention, where P <0.01 compared to model group is represented.

Detailed Description

Example 1

Taking 3kg of dried platycodon grandiflorum leaves, crushing, adding 70% ethanol in an amount which is 3 times that of the crushed platycodon grandiflorum leaves, performing reflux extraction for 3 times and 2 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, performing 200-mesh silica gel column chromatography on an ethyl acetate extract repeatedly, wherein an eluent is petroleum ether: ethyl acetate 80: 1; after preparative HPLC, the eluent was methanol: and (3) keeping the compound with the retention time of 60min when the water is 10:1 to obtain 14.5mg of platycodon grandiflorum neoalcohol.

Structure identification is carried out by adopting a spectroscopy method:

HR-ESI-MS gives the peak of the excimer ion M/z 315.1451[ M + HCOO]^-(theoretical value: 315.1444), and determining the molecular formula as C by combining NMR spectrum data₁₄H₂₂O₅

¹H-NMR(500MHz,C₅D₅N) gives a total of 17 proton signals, with 1 methyl proton signal given at high field: δ 1.20(3H, d, J ═ 5.3 Hz); the midfield gives the hydrogen signal on 6 vicinal oxygen carbons: 4.21(1H, d, J ═ 3.85Hz), 3.57(1H, m), 3.35(1H, m), 3.43(1H, m),3.99(1H, m), 3.74(1H, m); the hydrogen spectrum also gives 10 methylene hydrogen signals.

DEPTQSP(125.8MHz,C₅D₅N) gave a total of 14 carbon signals. Wherein 9 carbon signals are given in the middle field, and 4 quaternary carbon signals are identified by combining HMQC and HMBC spectrums: δ 66.8, 71.0, 76.1, 78.6, further presumed to be an alkyne carbon signal by literature reports, the remaining 5 being vicinal carbon signals: δ 68.2, 69.6, 71.3, 73.3, 80.3; giving 5 saturated carbon signals at high field: δ 18.5, 24.6, 28.4, 24.3, 27.1.

From the HMQC spectrum, 10 carbon signals and 17 proton signals were assigned, and 1 methyl carbon signal, 5 methylene carbon signals and 4 vicinal oxymethylene carbon signals were deduced.

In addition, in the HMBC spectrum, a methyl proton signal with a chemical shift of 1.20 is remotely correlated with δ 68.2, 71.3 two vicinal oxymethylene groups, and the hydrogens on the two vicinal oxymethylene groups are remotely correlated with 4 alkyne carbon signals, while two proton signals of δ 3.74(1H, m) and 4.21(1H, d, J ═ 3.85Hz) are also remotely correlated with a carbon signal with a chemical shift of 18.5; both hydrogens with chemical shifts δ 2.49 and 2.59 are correlated remotely with 4 alkyne carbon signals; the two hydrogen signals at δ 1.55 and 1.88 are remotely correlated with the two vicinal oxygen carbon signals at δ 69.6, 80.3; the hydrogen signal of delta 1.53 is remotely correlated with the oxygen-linked carbon signal of delta 80.3 and 73.3, other remotely correlated signals are also shown in the HMBC spectrum, the main remote correlation of the compound is shown in figure 1, and the HMBC spectrum is shown in figure 6.

Combine the above¹H-NMR, DEPTQSP, HMQC and HMBC spectra further belong to chemical shifts of carbon signals and hydrogen signals, and determine that the structure of the spectrum is 2,3,9,13,14-pentahydroxy-4,6-tetradecadiyne (2,3,9,13,14-pentahydroxy-4, 6-tetracadiyne). The information of the hydrocarbon attribution is shown in the table 1, and HMBC related signals, HR-ESI-MS spectrum,¹The H-NMR spectrum, DEPTQSP spectrum, HMQC spectrum and HMBC spectrum are shown in the attached figures 1-6 respectively.

Through comprehensive analysis and Scifinder search, the compound is not reported in the literature and is a new alkynol compound named platycodon neol (Platycodynol).

TABLE 1 preparation of platycodon neohydrin 1¹H-NMR and¹³C-NMR data

Example 2

Taking 3kg of dried platycodon grandiflorum leaves, crushing, adding 4 times of 80% ethanol for reflux extraction for 4 times, extracting for 3 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, subjecting an ethyl acetate extract to 200-mesh silica gel column chromatography repeatedly, wherein an eluent is petroleum ether: ethyl acetate 2: 1; after preparative HPLC, the eluent was methanol: the compound is retained for 45min at a water ratio of 1:1 to obtain 18.0mg of platycodon grandiflorum neoalcohol.

Example 3

Taking 3kg of dried platycodon grandiflorum leaves, crushing, adding 90% ethanol in an amount which is 5 times that of the crushed platycodon grandiflorum leaves, performing reflux extraction for 5 times and 4 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, subjecting an ethyl acetate extract to repeated 400-mesh silica gel column chromatography, wherein an eluent is petroleum ether: ethyl acetate 1: 20; after preparative HPLC, the eluent was methanol: and (3) taking the compound with the retention time of 30min as water at a ratio of 1:10 to obtain 9.7mg of platycodon grandiflorum neoalcohol.

Example 4

Taking dried platycodon grandiflorum leaves, crushing, adding 5 times of 80% ethanol for reflux extraction for 4 times, extracting for 3 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, subjecting an ethyl acetate extract to repeated 200-mesh silica gel column chromatography, wherein an eluent is petroleum ether: ethyl acetate 1: 1; after preparative HPLC, the eluent was methanol: and (3) keeping the compound with the retention time of 45min when the water is 10:3 to obtain the platycodon grandiflorum neoalcohol.

The present invention can be further illustrated by the following antidepressant test examples.

First, experimental material

1. Laboratory apparatus

R201D model constant temperature water bath, rotary evaporator (shanghai yukang scientific instruments ltd); an electronic balance of FA1104N type (precision scientific instruments ltd, national bridge, shanghai); FW177 type high-speed universal pulverizer (Guangming medical instruments, Beijing);

WB-07 mouse tail suspension device (self-made in laboratory, 25 cm. times.25 cm. times.35 cm).

2. Experimental Material

Platycodon grandiflorum neoalcohol: self-made in a laboratory, batch number: 20170919, respectively;

LPS: escherichia coli O127: B8, supplied by Sigma Co., lot number: 20171102, respectively;

fluoxetine hydrochloride: dalian Melam Biotechnology Ltd. Prazosin (alpha)₁-adrenoceptor antagonists), haloperidol (non-selective D)₂Receptor antagonists), bicuculline (competitive GABA antagonist), D-methyl-D-aspartate (NMDA, agonist of the glutamate site), purchased from Sigma company, usa.

3. Laboratory animal

The physical quality of the SPF-grade ICR male mice is 18-22 g, experiments are carried out by Yisi experimental animal technology Limited liability company in Changchun city, and the quality certification number is as follows: 201700020061. feeding the chickens in an environment with the room temperature of 22-24 ℃ and the relative humidity of 55-65%, and freely eating drinking water. The experimental protocol for all experimental mice of this study was approved by the ethical committee of the university of gilin.

Second, Experimental methods

1. Preparation of test article

Mixing ethanol extract of radix Platycodi neoalcohol with purified water to obtain high extract (20 mg. mL)^-1) Middle (10 mg. mL)^-1) Low (5 mg. mL)^-1) Solutions of 3 dose groups;

LPS was adjusted to a concentration of 0.83mg/kg with physiological saline^-1The solution of (4) is ready for use.

2. Grouping and administration of laboratory animals

In the 7 th day of adaptive feeding, 48 mice are screened out according to sugar water preference experiment, food intake and autonomic motion ability, and are randomly divided into 6 groups according to weight, wherein each group comprises 8 mice, and the experiments are specifically divided into the following groups:

normal control group (N): distilled water (1 mL. kg)^-1) Continuously performing intragastric administration for 7 days, and injecting physiological saline (0.83mg/kg) after intragastric administration for 0.5h on 7 days;

model control group (M): distilled water (1 mL. kg)^-1) Continuously performing intragastric administration for 7d, and injecting LPS after intragastric administration for 0.5h on 7 d;

positive drug fluoxetine group (F): fluoxetine (20mg kg)^-1) Continuously performing intragastric administration for 7d, and injecting LPS after intragastric administration for 0.5h on 7 d;

platycodon grandiflorum neo-alcohol administration group: high dose group (HR,400 mg. kg)^-1) Medium dose group (MR,200 mg. kg)^-1) And low dose group (LR,100 mg. kg)^-1) Continuously performing intragastric administration for 7d, and injecting LPS after intragastric administration for 0.5h at 7 d;

each group of test mice was tested by a study experiment 24h after LPS or normal saline injection. The mice were subjected to video acquisition and data analysis of the behavioural experiments simultaneously at the same time period. To avoid interference, all experiments were conducted in a quiet environment with background noise less than 65 db.

3. Sweet water preference experiment

Sugar water Preference Experiment (Sugar Preference Experiment): before the experiment, the mice are subjected to sugar water adaptability training for 3d, two water bottles of common water and 1% sucrose water are given to each mouse raised in a single cage, and the positions of the two water bottles are changed every 12 h. After the last administration for 1h, the same amount of common water and 1% sucrose solution are given to each group of mice, the consumption of the common water and the 1% sucrose solution in the mice within 24h is calculated, and the sugar solution preference rate is calculated according to the following formula:

the sugar water preference rate is equal to sugar water consumption/(sugar water consumption + common water consumption). times.100%

4. Body weight and food intake

The weight was weighed once every 2d and 24h after LPS injection, the change in body weight of the mice was recorded and calculated. Meanwhile, the food intake of the mice was calculated 24h after the LPS injection.

5. Open field experiment

Open Field Test (OFT): the mouse behavior experiment is also called an open box experiment and is a commonly used mouse ethology experiment, and the occurrence frequency, the duration and the like of certain behaviors of a mouse in a novel environment are used for reflecting the autonomous behavior and the exploration behavior of the mouse in a strange environment. The spontaneous activity of the mouse reflects the activity of the mouse, and the number of times of erection reflects the curiosity of the mouse on a fresh environment. The open field experiment device is a box with the height of 35cm and the length and the width of 72cm, and the bottom surface of the box is divided into 64 equal parts. The mice were placed in the median lattice, allowed to move autonomously for 5min, and their spontaneous activity (horizontal score, the number of squares traversed horizontally by the mice, i.e. the number of times the limbs cross the lattice) and the number of times of standing (vertical score, the two front paws of the mice vacate and are more than 1cm above the ground, or climb on the wall) were recorded. After each mouse is finished, the mouse is placed back into the cage, and the bottom and the four walls of the cage are cleaned by using 75% ethanol.

6. Forced swimming experiment

Forced Swimming Test (Forced Swimming Test, FST): a single mouse was placed in an organic glass jar (22 cm in height, 14cm in diameter, 12cm in depth of water, 25. + -. 1 ℃ C.) and subjected to a forced swimming test. The experiment was performed for 6min, the first 2min was the adaptation phase, and the immobility time after 4min was recorded, i.e. the mice stopped struggling, floated on the water surface, and had only limbs moved slightly to keep the head above the water surface. The mice were wiped dry and returned to the cage, each mouse requiring replacement of the water in the jar before being subjected to the forced swim test. Immobility time can reflect the despair status of the mice.

7. Tail suspension experiment

Tail overhang Test (TST): a single mouse is placed in a self-made tail suspension device, and the tail tip of the mouse is stuck to a cross rod in the tail suspension device at a position of about 2cm by using a medical adhesive tape. The experiment was carried out for 6min, the first 1min was the adaptation phase, and the immobility time of the last 5min was recorded, i.e. the mice stopped struggling to overcome abnormal body position, appeared with intermittent suspension immobility, and the time of disappointing state was displayed.

Third, experimental results

1. Sugar water preference rate

The sugar water preference rate for each group of mice in the sugar water preference experiment is shown in figure 7. The sugar water preference rate of the LPS-induced depression mice is significantly lower than that of the control group mice (P <0.01), indicating that the LPS-induced depression mice have a certain loss of interest; compared with the model group, the sugar water preference rate of the depression mice can be remarkably increased by the positive drug group (P <0.01), the medium dose group (P <0.01) and the high dose group (P <0.05), and the sugar water preference rate of the model group mice is not remarkably changed by other groups, which shows that fluoxetine, the medium dose platycodon neol and the high dose platycodon neol can improve the interest loss of the depression mice induced by LPS to a certain extent.

2. Body weight and food intake

The weight of the mice with the depression induced by LPS is obviously lower than that of the mice in a control group (P <0.01), which shows that the weight of the mice with the depression induced by LPS is obviously reduced, and the difference has statistical significance; compared with the model group, the positive drug group (P <0.01) and the medium dose group (P <0.01) can obviously increase the weight of the depression mice (see table 2 and figure 8). The fluoxetine and the platycodon grandiflorum neoalcohol with the medium dosage can improve the weight loss of mice with the LPS-induced depression to a certain extent.

The food intake of the mice with the LPS-induced depression is significantly lower than that of the mice in a control group (P <0.01), and the difference has statistical significance; compared with the model group, the positive medicine group (P <0.01) and the medium dosage group (P <0.01) can obviously increase the food intake of depression mice, and the results show that fluoxetine, platycodon grandiflorum leaves of each dosage group and platycodon grandiflorum roots of the medium dosage group can improve the decrease of the food intake of the depression mice induced by LPS to a certain extent, and the results are shown in figure 9.

Table 2 change in mouse body weight (mean ± SD, n ═ 8)

3. Open field experiment

The open field experiment mainly observes the autonomous behavior of the mice, and as can be seen from fig. 10A, the horizontal motor score of the LPS-induced depression mice is significantly reduced (P <0.05) compared with the normal group, which has statistical significance; compared with the model group, the positive medicine group (P <0.01) and the medium dose group (P <0.01) can increase the horizontal movement score of the model group to a certain extent; as can be seen from fig. 10B, LPS-induced depression mice had significantly reduced vertical motor scores (P <0.05) compared to the normal group, with statistical significance; compared with the model group, the positive medicine group (P <0.01) and the medium dose group (P <0.01) can increase the vertical movement score of the model group to a certain extent.

4 forced swimming test

The forced swimming test can judge whether the mice are depressed or not and the degree of depression, compared with a control group, the mice with the depression induced by LPS have longer immobility time (P <0.01), the behavior of depression is increased, and the statistical significance is achieved; compared with the model group, the immobility time of the positive drug group (P <0.01) and the medium dose group (P <0.01) can be obviously shortened, which shows that the immobility time of the depression mice is reduced along with the treatment of fluoxetine and platycodon neol, and the figure is 11.

5. Tail suspension experiment

The tail suspension test can also judge whether the mouse is depressed or not and the degree of depression, compared with a control group, the LPS-induced depression mouse has longer immobility time (P <0.01), has a certain degree of depression-like behavior and has statistical significance; compared with the model group, the immobility time can be obviously shortened by the positive medicine group (P <0.01) and the platycodon grandiflorum neol medium dosage group (P <0.01), which shows that the treatment of fluoxetine and platycodon grandiflorum neol can adjust the immobility time of mice to be normal level, and the figure 12 shows that the treatment of fluoxetine and platycodon grandiflorum neol can adjust the immobility time of mice to be normal.

The results show that: the platycodon grandiflorum neoalcohol has a remarkable anti-depression effect, can remarkably increase the weight and food intake of mice with depression induced by LPS (lipopolysaccharide), improve the sugar water preference rate of the mice, increase the horizontal score and the vertical times in an open field experiment of the mice, and remarkably shorten the immobility time in a forced swimming experiment and a tail suspension experiment of the mice.

Research on antidepressant action mechanism of platycodon grandiflorum neol

1. Experimental methods

The results of the behavioral experiments show that the platycodon grandiflorum neoalcohol has a remarkable antidepressant effect. The pathogenesis of the depression is unknown, but a large number of experimental studies show that the content change of monoamine neurotransmitters such as 5-hydroxytryptamine (5-HT), Norepinephrine (NA) and Dopamine (DA) is closely related to the depression. Therefore, we intend to search for an antidepressant mechanism by observing the influence of platycodon grandiflorum neoalcohol on 5-HT, NA and DA of hippocampus of a mouse model with LPS-induced depression. In the 7 th day of adaptive feeding, 48 mice are screened out according to sugar water preference experiment, food intake and autonomic motion ability, and are randomly divided into 6 groups according to weight, wherein each group comprises 8 mice, and the experiments are specifically divided into the following groups:

normal control group (N): distilled water (1 mL. kg)^-1) Continuously performing intragastric administration for 7 days, and injecting physiological saline (0.83mg/kg) after intragastric administration for 0.5h on 7 days;

platycodon grandiflorum neo-alcohol administration group: high dose group (HR,400 mg. kg)^-1) Medium dose group (MR,200 mg. kg)^-1) And low dose group (LR,100 mg. kg)^-1) Continuously performing intragastric administration for 7d, and injecting LPS after intragastric administration for 0.5h at 7 d;

prazosin group: prazosin (1mg/kg) was given 60 minutes prior to the experiment. Platycodon grandiflorum neohydrin (10mg/kg)

+ prazosin (1mg/kg) group: prazosin was administered 30min before the administration of platycodon neol and the behavioural test was performed 60min later.

Haloperidol group: haloperidol (0.2mg/kg) was administered 60 minutes prior to the experiment. Platycodon grandiflorum neo-alcohol (10mg/kg) + haloperidol (0.2 mg/kg): haloperidol was administered 30min before the administration of platycodon neoalcohol, and the behavioural experiment was performed 60min later.

Dicentrine group: bicuculline (4mg/kg) was administered 60 minutes before the experiment. Platycodon grandiflorum neoalcohol (10mg/kg) + dicentrine (4 mg/kg): dicentrine was administered 30min before the administration of platycodon grandiflorum neo-alcohol, and the behavioural test was performed 60min later.

NMDA group: NMDA (75mg/kg) was administered 60 minutes prior to the experiment. Campanulanol (10mg/kg) + NMDA (75 mg/kg): NMDA was administered 30min before neoalcohol in platycodon grandiflorum and behavioral experiments were performed 60min later.

2. Results of the experiment

The results show that: the platycodon grandiflorum neol has remarkable antidepressant effect, and the antidepressant effect of the platycodon grandiflorum neol can be related to a dopamine system, a GABA (gamma-aminobutyric acid) system, a glutamic acid system and a norepinephrine system, and is shown in figures 13A-D.

Claims (4)

1. A platycodon grandiflorum new alcohol: characterized by the following formula:

the molecular formula is as follows: c₁₄H₂₂O₅Molecular weight: 270.1476, respectively;

chemical name: 2,3,9,13,14-pentahydroxy-4, 6-tetradecadiyne;

brown oil, easily soluble in methanol, ethanol, pyridine, TLC detection, 10% H₂SO₄The ethanol solution is a color developing agent, and the color is blue when the mixture is heated at 105 ℃.

2. The extraction method of platycodon grandiflorum neo-alcohol according to claim 1, characterized by comprising the steps of:

taking dry platycodon grandiflorum leaves, crushing, adding 70-90% ethanol in an amount which is 3-5 times that of the crushed platycodon grandiflorum leaves, performing reflux extraction for 3-5 times, performing reflux extraction for 2-4 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, performing 200-mesh 400-mesh silica gel column chromatography on the ethyl acetate extract repeatedly, wherein an eluent is petroleum ether: ethyl acetate (80-1): (1-20); by preparative HPLC, eluent methanol: water (10-1): (1-10), and keeping the compound with the retention time of 30-60 min to obtain the platycodon grandiflorum new alcohol.

3. The extraction method of platycodon grandiflorum neo-alcohol according to claim 2, characterized by comprising the steps of:

taking dried platycodon grandiflorum leaves, crushing, adding 5 times of 80% ethanol for reflux extraction for 4 times, extracting for 3 hours each time, recovering under reduced pressure to obtain an extract, suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, discarding a petroleum ether layer and a dichloromethane layer, subjecting an ethyl acetate extract to repeated 200-mesh silica gel column chromatography, wherein an eluent is petroleum ether: ethyl acetate 1: 1; by preparative HPLC, eluent methanol: and (3) keeping the compound with the retention time of 45min when the water is 10:3 to obtain the platycodon grandiflorum neoalcohol.

4. Use of platycodon neohydrin according to claim 1 in the preparation of an antidepressant medicament.

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