CN111840258A

CN111840258A - Application of cis-methyl isoeugenol in preparing anti-depression drug

Info

Publication number: CN111840258A
Application number: CN202010811759.3A
Authority: CN
Inventors: 颜露; 韦敏; 宋萍萍; 钱怡云; 郑生智
Original assignee: Institute of Botany of CAS
Current assignee: Institute of Botany of CAS
Priority date: 2020-08-13
Filing date: 2020-08-13
Publication date: 2020-10-30

Abstract

The invention discloses an application of cis-methyl isoeugenol in preparing an anti-depression medicament. The cis-methyl isoeugenol has obvious effect on treating depression. The invention relates to an oral administration drug which is prepared from cis-methyl isoeugenol and is used for treating depression.

Description

Application of cis-methyl isoeugenol in preparing anti-depression drug

Technical Field

The invention belongs to the technical field of medicines, relates to medical application of cis-methyl isoeugenol, and particularly relates to application of cis-methyl isoeugenol in a medicine for effectively treating depression.

Background

Depression is a type of mood disorder characterized primarily by marked and persistent mood swings, and is one of the four major diseases in the world. The prevalence has increased globally in recent years, and has become the second largest disease in humans after coronary heart disease. The incidence of depression in China is about 4%, depression patients cannot adapt to the society, and the health-care depression-treatment pillow is a high-risk group with suicide, self-disability and self-abuse and constitutes a serious health crisis for the society. Clinical studies find that depression is also a complication of various diseases such as parkinson's disease, diabetes and the like, and a serious neuropsychiatric burden is brought to patients. Therefore, depression has become a prominent social problem, and the research on depression has become a hot spot of concern at home and abroad.

Modern researches indicate that the pathogenesis of depression is complex and is related to factors such as heredity, environment, society and the like. The following hypotheses are mainly involved: 1 neurotransmitter hypothesis, central 5-hydroxytryptamine nervous system dysfunction, 5-hydroxytryptamine (Serotonin, 5-HT) in synaptic cleft, Norepinephrine (NE), and Dopamine (DA) levels are among the major pathogenesis of depression. 2, the neurotrophic factor hypothesis, Nerve Growth Factor (NGF), Brain-derived neurotrophic factor (BDNF), Glial cell line-derived neurotrophic factor (GDNF), etc. have important roles in the growth and differentiation of neurons and the formation of neural circuits. The expression level of the hippocampal neurotrophic factor is reduced, neurogenesis and neuron survival are influenced, and depression symptoms are caused; the expression of the homeopathic nutrition factors can be regulated, and the depression symptoms can be relieved. 3, under the neuroinflammation hypothesis, the level of central inflammatory factors such as Interleukin 1 beta (Interleukin-1 beta, IL-1 beta), Interleukin 6 (Interleukin-6, IL-6), Tumor necrosis factor-alpha (Tumor necrosis factor-alpha, TNF-alpha) and the like is increased, neurotransmitters, neurotrophic factors or nerve circuits related to the emotional modulation are influenced, and depression symptoms are generated; reducing the expression of inflammatory factors and relieving depression symptoms.

Clinically, the increase of synaptic cleft neurotransmitter level is the main strategy for treating depression. The anti-depressant drugs are mainly represented by: tricyclic antidepressants (TCAs), such as imipramine, and the like; selective Serotonin Reuptake Inhibitors (SSRIs), such as fluoxetine, and the like; monoamine oxidase inhibitors (MAOI), such as phenelzine, and the like. These drugs have various side effects and high tolerability, and there is a delay in treatment time, suggesting that it is difficult to achieve good effects for the prevention and treatment of depression only by regulating neurotransmitter levels. The neurotrophic factor and neuroinflammation hypothesis think that the onset of depression is closely related to inflammation, neurogenesis and the like, reduces the expression of inflammatory factors, increases the expression of neurotrophic factors, improves inflammation and neurogenesis, can effectively relieve depression, and is also helpful for explaining that the antidepressant needs a plurality of weeks to improve the inflammation and neurogenesis to play a curative effect. Therefore, neurotransmitter, neurotrophic factors and inflammatory factors are used as targets, and new antidepressant drugs are searched for to effectively prevent and treat the depression.

cis-Methyl isoeugenol (cis-Methyl isoeugenol) is a phenylpropanoid compound and is mainly distributed in Acorus tatarinowii Schott of Araceae, but the main active ingredients of the Acorus tatarinowii Schott are beta-asarone, alpha-asarone and Methyl eugenol, and the cis-Methyl isoeugenol has less content in the Acorus tatarinowii Schott. The cis-methyl isoeugenol is oily liquid, and has a molecular formula: C11H14O2, molecular weight: 178.23 has the following structure, and has effects of tranquilizing mind, improving sleep, and improving memory. At present, the application of cis-methyl isoeugenol in preparing the anti-depression medicament is not found.

Disclosure of Invention

The invention aims to solve the technical problem of providing the application of cis-methyl isoeugenol in preparing anti-depression drugs. The cis-methyl isoeugenol has the following structure:

the invention relates to an application of cis-methyl isoeugenol in resisting depression, which can obviously improve the level of neurotransmitter, up-regulate the psychotrophic factor and down-regulate the expression of inflammatory factor. Therefore, the cis-methyl isoeugenol can be applied to the preparation of anti-depression drugs, and the drugs have the application of treating depression.

Cis-methyl isoeugenol can be obtained from the market or prepared by adopting the prior art.

The cis-methyl isoeugenol applied by the invention can be prepared into a medicament with any pharmaceutically allowed auxiliary material or pharmaceutical excipient, and the preparation can be any pharmaceutically allowed dosage form, including but not limited to granules, tablets, medicinal granules, soft capsules or dropping pills.

The administration form of the medicament provided by the invention mainly comprises oral administration and the like.

The dosage of cis-methyl isoeugenol in the present invention varies depending on the condition, body weight, administration mode and the like of a patient. For example, the dosage is 0.1-5 mg/(kg.d) for intramuscular or enteral administration in the case of non-oral administration, and 0.4-20 mg/(kg.d) for oral administration.

Compared with the prior art, the invention has the beneficial effects that: the cis-methyl isoeugenol can obviously shorten the forced swimming and tail suspension accumulation immobility time of normal mice, has obvious antidepressant effect on chronic stress depressed mice, can obviously improve the 5-hydroxytryptamine, norepinephrine, 5-oxindole acetic acid/5-hydroxytryptamine level and striatal dopamine level of the hippocampus of depressed mice, and can up-regulate neurotrophic factors of the hippocampus and down-regulate the expression of inflammatory factors. Therefore, the cis-methyl isoeugenol can be applied to the preparation of the anti-depression drug. The invention provides a new clinical application of cis-methyl isoeugenol and enlarges the application range of the cis-methyl isoeugenol.

Drawings

FIG. 1 is a graph showing the results of a forced swimming test of a mouse.

FIG. 2 is a graph showing the results of the tail suspension test of mice.

FIG. 3 is a graph of the results of weight measurements of chronically depressed stress mice.

FIG. 4 is a graph showing the results of the carbohydrate preference test in chronic stress-depressed mice.

FIG. 5 is a graph showing the results of a forced swimming test for chronic stress-depressed mice.

FIG. 6 is a graph showing the results of neurotransmitter detection in chronically stressed depressed mice.

FIG. 7 is a graph of the result of detecting neurotrophic factor in hippocampal region of mice with chronic stress depression.

FIG. 8 is a graph showing the result of detecting inflammatory factors in hippocampal region of chronic stress depressed mice.

Detailed Description

The present invention will be further described with reference to specific examples, which are provided for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental animals are purchased from Changzhou Kavens experimental animals Limited company (production license number: SCXK (Su) 2016-. The cis-methyl isoeugenol reference substance is prepared by self, has the purity of more than or equal to 98 percent and is suspended in water to be prepared into the administration concentration (3.3, 10, 30 mg/mL).

EXAMPLE one Effect of cis-methyl Isoeugenol on cumulative immobility time in forced swimming of mice

ICR mice, male, were bred for 7 days to adapt to the new environment. Mice were divided into a blank control group, a cis-methylisobutyrin group [16.6, 50, 150 mg/(kg.d) ], an imipramine group [30 mg/(kg.d) ], a levodopa group [60 mg/(kg.d) ], and a galantamine group [1.5 mg/(kg.d) ], 12 mice each, administered by gavage at 10:00 a.m.for 7 days. After 1h of the last administration, a forced swimming test was performed. The test was performed in 2 days: the mice were allowed to swim in 22 + -2 deg.C deep water for 15min on day 1, and after 24 h, forced swimming was performed for 6 min under the same conditions, and the cumulative immobile time for swimming within 4min after counting (i.e., the mice stopped struggling or the mice were floating, and the limbs slightly moved to keep the head on the water surface) was shown in FIG. 1. The result shows that the accumulative immobility time of the forced swimming of the mouse after the positive drug and the cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ] are obviously different from the model group (p <0.05 or p < 0.01).

EXAMPLE two Effect of cis-methyl Isoeugenol on cumulative immobility time of mouse tail suspension

ICR mice, male, were bred for 7 days to adapt to the new environment. Mice were divided into a blank control group, a cis-methylisobutyrin group [16.6, 50, 150 mg/(kg.d) ], an imipramine group [30 mg/(kg.d) ], a levodopa group [60 mg/(kg.d) ], and a galantamine group [1.5 mg/(kg.d) ], 12 mice each, administered by gavage at 10:00 a.m.for 7 days. The tail suspension test was performed 1h after the last administration. The position about 1 cm away from the tail tip of the mouse is pasted on a bracket of a tail suspension box (25 multiplied by 30 cm) by using an adhesive tape to enable the mouse to be in an upside-down suspension state, the suspension time is 6 min, the accumulated dead time of tail suspension in the 6 min of the mouse is counted (namely the mouse stops struggling or has no movement), and the result is shown in figure 2. The result shows that the cumulative immobility time of the tail suspension of the mice after the positive drug, cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ], levodopa and galantamine act is obviously different from that of the model group (p is less than 0.05 or p is less than 0.01).

EXAMPLE III Effect of cis-methyl Isoeugenol on body weight in chronically stress depressed mice

C57/BL6 mice were used, male, and kept for 7 days to adapt to the new environment. Except for the placebo group, animals were randomized to the following series of chronic stress stimuli: (1) 2 h of noise; (2) the body is bound for 1 h; (3) wet cage for 15 h (200 mL water in cage); (4) cage tilt (45 °) 15 h; (5) stroboscopic for 2 h; (6) fasting for 6 h; (7) forced swimming for 6 min at 8 deg.C; (8) suspending tail for 6 min; (9) the day and night are reversed. The above stress stimulation was randomly scheduled to be completed within one week, and repeated for 6 weeks to establish a chronic stress depression model. The model mice were divided into a model group, a cis-methyl isoeugenol group [16.6, 50, 150 mg/(kg.d) ], an imipramine group [30 mg/(kg.d) ], a levodopa group [60 mg/(kg.d) ], and a galantamine group [1.5 mg/(kg.d) ], 12 mice each, administered by gavage at 10:00 a.m.daily for 6 weeks. Body weight measurements were made for each group of mice after the modeling was completed and the dosing was completed, and the results are shown in fig. 3. The results show that the model group has significant difference (p < 0.01) with the normal control group, and the body weight has significant difference (p < 0.05) with the model group after 6 weeks of filling positive drug, levodopa and cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ].

[ example IV ] Effect of cis-methyl Isoeugenol on sugar Water preference in Chronic stress Depression mice

Sugar water preference = sucrose water consumption/total liquid consumption x 100%.

After dosing was complete, 72 h before the sugar bias test, mice were trained to accommodate a 1% aqueous sucrose solution (weight/volume): 2 bottles of 1% sucrose in water were placed in each cage and after 24 h, 1 of these bottles was changed to drinking water for 24 h. After acclimation training was completed, the mice were deprived of water and food for 24 h. The sugar water bias test was started at 9:00 am, 1 mouse per cage, and the mice were free to choose 2 water bottles (containing 100 mL sucrose solution (1%, weight/volume) and 100 mL drinking water, respectively). After 3 h, the volume of sucrose and drinking water consumed by the mice was recorded. The calculation formula of the sugar water preference degree is as follows: sugar water preference =1% sucrose water consumption volume/(water consumption volume +1% sucrose water consumption volume) × 100%, the results are shown in fig. 4. The results show that the model group has significant difference (p < 0.01) with the normal control group, and the sugar water preference degree after 6 weeks of filling the positive drug and cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ] has significant difference (p < 0.05) with the model group.

EXAMPLE five Effect of cis-methyl Isoeugenol on cumulative immobility time in forced swimming of chronically stressed depressed mice

After completion of the administration, a forced swimming test of mice was performed. The test was performed in 2 days: the mice were allowed to swim in 22 + -2 deg.C deep water for 15min on day 1, and after 24 h, forced swimming was performed for 6 min under the same conditions, and the cumulative immobile time of swimming within 4min after counting (i.e., the mice stopped struggling or the mice were floating, and the limbs slightly moved to keep the head on the water surface) was counted, and the results are shown in FIG. 5. The results show that the model group has significant difference (p < 0.01) from the normal control group, and the cumulative immobility time of forced swimming after 6 weeks of positive drug infusion and cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ] has significant difference (p <0.05 or p < 0.01) from the model group.

EXAMPLE six Effect of cis-methyl Isoeugenol on neurotransmitter levels in chronically depressed stress mice

After behavioral assessment, animals were sacrificed and samples of hippocampal, striatal tissue were taken and tested for norepinephrine, 5-hydroxytryptamine, 5-oxindole acetic acid/5-hydroxytryptamine, and dopamine levels. The sample was treated with tissue lysate (0.6 mol/L perchloric acid, 0.5 mmol/LNa)₂EDTA, 0.1 g/L L-cysteine mixed aqueous solution), and then freezing and centrifuging to obtain supernatant; adding perchloric acid precipitant (1.20 mol/L K)₂HPO₄、2.00 mmol/L Na₂Mixed aqueous solution of EDTA), refrigerated centrifugation, and filtration. The chromatographic conditions and fluorescence detection parameters were as follows: agilent HPLC 1260, Shim-pack C18 column (250 mm. times.4.6 mm, 5 μm) (Shimadzu Japan); the mobile phase is citric acid-sodium acetate buffer (50 mmol/L citric acid, 50mmol/L sodium acetate, 0.5 mmol/L sodium 1-heptanesulfonate, 5 mmol/L triethylamine, 0.5 mmol/L Na₂EDTA) -methanol (83: 13, v/v) (pH 8.3); the flow rate is 1.0 mL/min; the sample injection amount is 10 mu L; the emission wavelength was 330 nm and the excitation wavelength was 280 nm, the results are shown in FIG. 6. The results showed that the model group was significantly different from the normal control group (p)<0.05 or p<0.01), drench cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d)]After 6 weeks the levels of 5-hydroxytryptamine, norepinephrine and 5-oxindole acetic acid/5-hydroxytryptamine and dopamine differed significantly from the model group (p)<0.05 or p<0.01）。

EXAMPLE seventy-The Effect of cis-methyl Isoeugenol on Hippocampus neurotrophic factor expression in chronically stressed depressed mice

After behavioral assessment, animals were sacrificed and hippocampal tissue samples were taken and tested for expression levels of hippocampal nerve growth factor, brain-derived neurotrophic factor and glial cell-derived neurotrophic factor. Extracting total RNA from the sample by using an RNA separation kit, and detecting the RNA content by using Nanodrop. A1 mu gRNA sample is taken, is subjected to reverse transcription by a reverse transcription kit to form cDNA, and a real-time quantitative PCR test is carried out by adopting an SYBR Green I chimeric fluorescence method, and the result is shown in figure 7. The results show that the model group has significant difference (p <0.05 or p < 0.01) with the normal control group, and the expression level of the neurotrophic factor after 6 weeks of positive drug infusion and cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ] has significant difference (p < 0.05) with the model group.

EXAMPLE VIII Effect of cis-methyl Isoeugenol on Hippocampus inflammatory factor expression in chronically depressed stress mice

After behavioral assessment, animals were sacrificed and hippocampal tissue samples were taken and tested for expression levels of interleukin-1 β, interleukin-6 and tumor necrosis factor- α in hippocampal regions. Extracting total RNA from the sample by using an RNA separation kit, and detecting the RNA content by using Nanodrop. A1 mu gRNA sample is taken, is subjected to reverse transcription by a reverse transcription kit to form cDNA, and a real-time quantitative PCR test is carried out by adopting an SYBR Green I chimeric fluorescence method, and the result is shown in figure 8. The results show that the model group has significant difference (p < 0.01) with the normal control group, and the expression level of the inflammatory factor has significant difference (p <0.05 or p < 0.01) with the model group after 6 weeks of positive drug infusion and cis-methyl isoeugenol [16.6, 50, 150 mg/(kg.d) ].

EXAMPLE nine cis-methyl Isoeugenol Capsule

100 g of cis-methyl isoeugenol, 80 g of starch, 24 g of sodium carboxymethyl starch, 17 g of dextrin and a proper amount of 70% ethanol, and the mixture is prepared into 1000 capsules (each capsule contains 100 mg of cis-methyl isoeugenol) according to a conventional method.

[ example ten ] cis-methyl Isoeugenol Capsule

15 g of cis-methyl isoeugenol, 10 g of starch, 4 g of sodium carboxymethyl starch, 12 g of dextrin and a proper amount of 70% ethanol, and the mixture is prepared into 1000 capsules (each capsule contains 15 mg of cis-methyl isoeugenol) according to a conventional method.

EXAMPLE eleven cis-methyl Isoeugenol tablet

20 g of cis-methyl isoeugenol, 6 g of lactose, 3 g of compressible starch, 3 g of hydroxypropyl cellulose, 1 g of magnesium stearate and a proper amount of 70% ethanol, and the mixture is prepared into 1000 tablets (each tablet contains 20 mg of cis-methyl isoeugenol) by a conventional tabletting method.

EXAMPLE twelve cis-methyl Isoeugenol capsules

20 g of cis-methyl isoeugenol, 13 g of starch, 5 g of sodium carboxymethyl starch, 5 g of dextrin and a proper amount of 70% ethanol, and the capsule is prepared into 1000 capsules (each capsule contains 20 mg of cis-methyl isoeugenol) by a conventional method.

[ EXAMPLE thirteen ] cis-methyl Isoeugenol Capsule

40 g of cis-methyl isoeugenol, 17 g of starch, 17 g of sodium carboxymethyl starch, 5 g of dextrin and a proper amount of 70% ethanol, and the capsule is prepared into 1000 capsules (each capsule contains 40 mg of cis-methyl isoeugenol) by a conventional method.

[ example fourteen ] cis-methyl Isoeugenol tablets

150 g of cis-methyl isoeugenol, 55 g of lactose, 15 g of compressible starch, 10 g of hydroxypropyl cellulose, 5 g of magnesium stearate and a proper amount of 70% ethanol, and 1000 tablets (each tablet contains 150 mg of cis-methyl isoeugenol) are prepared according to a conventional tabletting method.

[ example fifteen ] cis-methyl Isoeugenol granules

6 g of cis-methyl isoeugenol, 800 g of lactose powder, 194 g of magnesium stearate and a proper amount of 70% ethanol, and the granules are prepared into 1000 g of granules (each g of granules contains 6 mg of cis-methyl isoeugenol).

EXAMPLE sixteen cis-methyl Isoeugenol Soft capsules

20 g of cis-methyl isoeugenol, 10 g of soybean oil, 5 g of gelatin, 5 g of glycerol and 7 g of distilled water, and the raw materials are prepared into 1000 granules (each granule contains 20 mg of cis-methyl isoeugenol) according to the preparation method of the conventional soft capsule.

Claims

1. Application of cis-methyl isoeugenol in preparing antidepressant is provided.

2. The use according to claim 1, wherein said cis-methyl isoeugenol is used to increase neurotransmitter levels, up-regulate psychotrophic factors and down-regulate inflammatory factor expression.

3. The use of claim 1, wherein the cis-methyl isoeugenol is combined with any pharmaceutically acceptable adjuvant or pharmaceutical excipient to form any pharmaceutically acceptable pharmaceutical dosage form.

4. The use as claimed in claim 3, wherein the dosage form includes granules, tablets, granules, soft capsules, soft capsules or dripping pills.

5. The use according to claim 3, wherein the medicament comprises 0.1 to 1200 mg of cis-methyl isoeugenol.