CN103142629A - Application of neohesperidin as antidepressant drug - Google Patents
Application of neohesperidin as antidepressant drug Download PDFInfo
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Abstract
The invention discloses application of neohesperidin as an antidepressant drug. Dosage forms of the drug comprise pharmaceutically acceptable dosage forms, such as tablets, capsules, solutions, suspensions, injections and infusion solutions. The drug consists of neohesperidin and pharmaceutically acceptable aglycone and secondary glycoside of neohesperidin which are produced through hydrolysis or enzymolysis and comprises various drug dosage forms which are prepared in a manner that the drug is combined with pharmaceutical carriers. The drug has little toxicity and is free from the disadvantages of conventional antidepressant drugs, such as excited anxiety, constipation, anuresis, dizziness, sleepiness and tremor.
Description
Technical field:
The present invention relates to medical technical field, exactly it is that neohesperidin is as the application of antidepressant drug.
Background technology:
Neohesperidin (Neohesperidin) is the flavanone glycosides compound, for Rutaceae citrus plant secondary metabolite, is distributed widely in a lot of plants.Studies show that, neohesperidin has stronger biological activity at aspects such as promoting gastric motility, blood fat reducing, atherosclerosis, blood sugar lowering, promotion skin microcirculation.the patent No.: CN1836665B discloses the patent of invention that name is called " pharmaceutical applications of neohesperidin or its compositions ", the patent No.: CN1317938A discloses the patent of invention that name is called " be used for the treatment of or prevention and blood fat and blood sugar level raise the compositions that contains neohesperidin pair hydrogen chalcones of relevant disease ", the patent No.: CN102258528A discloses the patent of invention that name is called " the new purposes of neohesperidin ", neohesperidin is disclosed in treatment or prevention disease such as the hyperlipidemia relevant to blood fat or blood sugar level rising, arteriosclerosis, angina pectoris, apoplexy, flatulence, dyspepsia, infantile anorexia, improve and/or promote the application of skin microcirculation.Have no the new purposes of neohesperidin in preparation anti-depression aspect medicine.
Summary of the invention:
The invention provides the antidepressant new purposes of neohesperidin.Neohesperidin can be used for prevention or treatment anxiety state or depressive state.
Neohesperidin comprises and the pharmaceutical carrier combination, can be used for prevention or treatment depressive state.Wherein said sick body is mammal.Neohesperidin can be by oral, non-intestinal or topical routes, and form of administration can be the pharmaceutically acceptable dosage forms such as tablet, capsule, solution, suspension, injection, drip liquid, lyophilized powder.
Advantage of the present invention is: the present invention has disclosed neohesperidin and has had antidepressant effect, for its application provides new approach.And toxicity is little, without the excited uneasiness of conventional antidepressants, constipation, urine retention, dizzy, drowsiness, the shortcoming such as tremble.
The specific embodiment:
Below zoopery be to further description of the present invention, but and do not mean that any limitation of the invention.
Example 1. autonomic activities irritability experiments
1. laboratory animal
The Wistar rat, male, body weight 200～220g, the SPF grade standard is weighed animal, and numbering is selected 30 of healthy rats.By the sequence of body weight size, be divided into 3 groups with randomized blocks, 10 every group.If blank group, positive controls, neohesperidin administration group.
2. sample source and processing
(1) blank group: normal saline, NaCl content 0.9%.
(2) positive controls: get fluoxetine Hydrochloride and use the normal saline standardize solution in volumetric flask, fluoxetine Hydrochloride concentration is 1.8mg/ml.Dosage is 7.2mg/kg.
(3) neohesperidin administration group: get neohesperidin appropriate, in volumetric flask, neohesperidin concentration is 1.6mg/ml with the normal saline standardize solution.Neohesperidin sample purity used is more than 95%, and dosage is 8mg/kg.
3. experimental technique (open field test)
Adopting the bottom surface is foursquare spacious case, long 100cm, and wide 100cm, high 50cm, the bottom surface is divided into close fan-shaped of 100 areas with black line.During experiment, rat is placed on spacious case center, gets over lattice number (at least 3 pawls stride across the border) with horizontal movement in rat 5min and estimate its mobility; Estimate its inquiry with the upright number of times that moves both vertically (two fore paw built on stilts).Whether have autonomous excitation, simultaneously with the contrast of accompanying of the blank group that gives normal saline and the positive controls that gives the positive control drug fluoxetine Hydrochloride if investigating neohesperidin by autonomic movement situation of change before and after each administration group administration relatively.
3. experimentation
By observing the activity of rat in spacious case, record the interior horizontal movement of 5min and get over the lattice number and the upright number of times that moves both vertically before administration; After 7 days, observe the activity of rat in spacious case in administration, record the interior horizontal movement of 5min and get over the lattice number and the upright number of times that moves both vertically.Each group experimental data is processed with the SPSS statistical software, and the horizontal movement rate of change is as follows with the rate of change computational methods that move both vertically:
Before horizontal movement rate of change=(administration after 7 days horizontal movement more horizontal movement lattice number more before lattice number-administration)/administration, lattice number * 100% is got in horizontal movement
More lattice number * 100% that moves both vertically before rate of change=(administration move both vertically after 7 days the more lattice number that moves both vertically before more lattice number-administration)/administration moves both vertically
Each group experimental data is carried out the t check, investigate it and whether have significant difference.
4. experimental result
By statistics, each administration group rat autonomic movement situation sees Table 1.
Before table 1. administration with administration each administration group rat open field test autonomic movement situation of change after 7 days
Use SPSS software, carry out the t check, autonomic movement situation before and after more blank group respectively, neohesperidin group and the administration of fluoxetine Hydrochloride group, the same with positive control drug fluoxetine Hydrochloride group with the blank group, the rat horizontal movement of neohesperidin administration group and the difference (P＞0.05) that moves both vertically that there are no significant before and after the administration show after giving neohesperidin the central nervous system without autonomous excitation.
Example 2. chronic stress rat open field tests
1. laboratory animal
The Wistar rat, male, body weight 200～220g, the SPF grade standard is weighed animal, and numbering is selected totally 40 of healthy rats.By the sequence of body weight size, be divided into 4 groups with randomized blocks, 10 every group.If blank group, pathological model group, positive controls, neohesperidin administration group.
2. sample source and processing
(1) blank group: with embodiment 1.
(2) pathological model group: normal saline, NaCl content 0.9%.
(3) positive controls: with embodiment 1.
(4) neohesperidin administration group: with embodiment 1.
3. experimental technique
(1) preparation of rat chronic Stress model: chronic stress stimulates as a kind of non-damage, with the process of mankind's psychosoma disease, similarity is arranged, the simulation that chronic stress model reality is tested " stress state " that run in people's daily life, for the specific clinical antidepressants of screening, the good suitability is arranged, this experiment is just take the chronic stress model as the basis, give positive control drug and neohesperidin, investigate the antidepressant effect of neohesperidin.
In the preparation of rat chronic Stress model, make stress stimulation meet the characteristics of unpredictability as far as possible, 28d is by the intensity of every day 1 or 2 kind of stimulation continuously, give alternately that rat is following to be stimulated: fasting 24h, prohibit water 24h, restriction and ingest that 2h, pairing raise 12h, the 45 ° of 2h of cage that incline, the 12h that throws light on all night, moist (200ml water is added to the 100g bedding and padding) 12h, stroboscopic (100min/s) 1h, white noise (110dB) 1h, forced swimming (4 ℃ of water temperatures) 5min, the braking 1h of raising, preparation chronic stress model.It is as follows that each administration group stress reach the administration situation:
A. blank group: do not stimulate, give normal saline.
B. pathological model group: stimulate to give simultaneously normal saline.
C. positive controls: stimulate to give simultaneously positive drug, dosage is with embodiment 1.
D. neohesperidin administration group: stimulate to give simultaneously neohesperidin, dosage is with embodiment 1.
(2) open field test: adopting the bottom surface is foursquare spacious case, long 100cm, and wide 100cm, high 50cm, the bottom surface is divided into close fan-shaped of 100 areas with black line.During experiment, rat is placed on spacious case center, gets over lattice number (at least 3 pawls stride across the border) with horizontal movement in rat 5min and estimate its mobility; Estimate its inquiry with the upright number of times that moves both vertically (two fore paw built on stilts).Investigate neohesperidin by neohesperidin administration group before and after administration relatively with pathological model group autonomic movement rate of change (comprising the horizontal movement rate of change and the rate of change that moves both vertically) and whether have antidepressant effect, the while is with the contrast of accompanying of the blank group that gives normal saline and the positive controls that gives the positive control drug fluoxetine Hydrochloride.
4. experimentation
By observing the activity of rat in spacious case, record the interior horizontal movement of 5min and get over the lattice number and the upright number of times that moves both vertically before administration; After 28 days, observe the activity of rat in spacious case at chronic stress, record the interior horizontal movement of 5min and get over the lattice number and the upright number of times that moves both vertically.Horizontal movement rate of change and the rate of change computational methods that move both vertically are as follows:
The horizontal movement rate of change=(stress be after 28 days horizontal movement more the lattice number-stress before horizontal movement lattice number more)/stress before horizontal movement get over lattice number * 100%
Move both vertically rate of change=(the lattice number that stress move both vertically after 28 days more-stress before the more lattice number that moves both vertically)/stress before more lattice number * 100% that moves both vertically
Result obtains positive number for increasing, and negative is for descending, and each group experimental data is processed with the SPSS statistical software, carries out the t check, investigates neohesperidin administration group and pathological model group autonomic movement situation and whether has significant difference.
5. experimental result
By statistics, each administration group autonomic movement situation sees Table 2.
Each administration group open field test autonomic movement situation of change of table 2 chronic stress rat model
Use SPSS software, carry out the t check, relatively there is extremely significant difference (P＜0.01) in the pathological model group with blank group autonomic movement situation (horizontal movement rate of change and the rate of change that moves both vertically) before and after administration, show under the state of chronic stress, rat do not have under pharmaceutical intervention its autonomic movement emotionally condition be subject to impact, horizontal movement rate of change and the rate of change that moves both vertically all obviously descend, and prompting chronic stress model is successfully prepared; Each administration group and pathological model group autonomic movement situation before and after administration compares (horizontal movement rate of change and the rate of change that moves both vertically) and all has extremely significant difference (P＜0.01), show that neohesperidin and positive control drug fluoxetine Hydrochloride all can have significant resistant function to the rat horizontal movement rate of change that causes due to chronic stress and the rate of change decline that moves both vertically, the prompting neohesperidin has the pharmacologically active of resisting the autonomic activities minimizing that causes due to depression.
Example 3. chronic stress rat body weight increment rate determination experiments
1. laboratory animal
With embodiment 2.
2. sample source and processing
(1) blank group: with embodiment 1.
(2) pathological model group: with embodiment 2.
(3) positive controls: with embodiment 1.
(4) neohesperidin administration group: with embodiment 1.
3. experimental technique
(1) preparation of rat chronic Stress model: the preparation of rat chronic Stress model and each administration group stress reach the administration situation all with example 2.
(2) body weight increment rate experiment: rat is after being stimulated by chronic stress, will the performance of loss of appetite appears, the decline that the minimizing of feed will cause body weight to be advanced the speed, therefore, by the situation of change with pathological model group body weight increment rate before and after administration of neohesperidin group relatively, can investigate neohesperidin and resist the impaired body weight that causes of digestive system function that causes due to the rat chronic stress stimulation decline of advancing the speed; Investigate the difference of neohesperidin group and pathological model group administration front and back respectively organizing the rat body weight increment rate and whether have significance, the positive controls that gives simultaneously the blank group of normal saline and give the positive control drug fluoxetine Hydrochloride contrast of accompanying.
4. experimentation
Rat body weight is respectively organized in weighing respectively before administration, and chronic stress stimulated after 28 days, and rat body weight is respectively organized in weighing again, respectively organizes the rat body weight increment rate before and after investigation neohesperidin group and the administration of pathological model group.The computational methods of rat body weight increment rate are as follows:
The body weight increment rate=(stress be after 28 days rat body weight-stress before rat body weight)/stress before rat body weight * 100%
Each group experimental data is processed with the SPSS statistical software, carried out the t check, investigate neohesperidin and pathological model group rat body weight increment rate and whether have significant difference.
5. experimental result
By statistics, each administration group rat body weight increment rate situation sees Table 3.
Each administration group rat body weight increment rate situation of change of table 3
Use SPSS software, carry out the t check, relatively there is extremely significant difference (P＜0.01) in the pathological model group with blank group body weight increment rate before and after administration, show under the state of chronic stress, under pharmaceutical intervention, its digestive system function has been subject to impact to rat not having, body weight is advanced the speed and is starkly lower than blank group, points out simultaneously the chronic stress model to be successfully prepared; More all there is extremely significant difference (P＜0.01) in neohesperidin group, positive control drug group and pathological model group body weight increment rate before and after administration, show that neohesperidin and positive control drug fluoxetine Hydrochloride all can advance the speed to descend to the rat body weight that causes due to chronic stress significant resistant function is arranged, the prompting neohesperidin has the not normal pharmacologically active of digestive system function that opposing causes due to depression.
Example 4. chronic stress rat sucrose are degree of having a liking for determination experiment partially
1. laboratory animal
With embodiment 2.
2. sample source and processing
(1) blank group: with embodiment 1.
(2) pathological model group: with embodiment 2.
(3) positive controls: with embodiment 1.
(4) neohesperidin administration group: with embodiment 1.
3. experimental technique
(1) preparation of rat chronic Stress model: the preparation of rat chronic Stress model and each administration group stress reach the administration situation all with example 2.
(2) sucrose water degree of having a liking for experiment partially: rat is after being stimulated by chronic stress, with the performance that interest occurs and go down, at first this experiment is carried out sucrose water to rat and is partially had a liking for training before carrying out chronic stress, set up it to the preference of sucrose solution, after carrying out the chronic stress stimulation, can cause because of the disappearance of interest the decline of degree of having a liking for partially of sucrose water for the rat of antidepressant drug, therefore, by relatively each administration group and pathological model group sucrose water degree of having a liking for situation of change partially before and after administration, can investigate the anhedonia that the neohesperidin antagonism causes due to the rat chronic stress stimulation; Investigate and respectively to organize rat sucrose water before and after neohesperidin group and the administration of pathological model group and partially whether have significance between degree of having a liking for rate of change, the while is with the contrast of accompanying of the blank group that gives normal saline and the positive controls that gives the positive control drug fluoxetine Hydrochloride.
4. experimentation
First rat is carried out learning training before administration: water is prohibited in fasting, only gives 1% sucrose water 48h; Sucrose degree of having a liking for is partially measured: after training end, normal drinking water is raised 3d, then fasting taboo water 23h, then allows rat freely drink 2 bottles of different water, and wherein one bottle is 1% sucrose water, and one bottle is tap water.Carry out the amount of drinking water (g) of 1h and measure, by formula calculate sucrose degree of having a liking for partially.Rat conform after 1 week by sucrose partially degree of having a liking for 40 rat equilibriums are divided into 4 groups: chronic stress is measured rat sucrose water degree of having a liking for partially after stimulating 28 days again.Calculate simultaneously sucrose degree of having a liking for rate of change partially
Sucrose is degree of having a liking for=sucrose diseases caused by retention of fluid consumption/(sucrose diseases caused by retention of fluid consumption+drinking purpose of tap water amount) * 100% partially
Sucrose partially degree of having a liking for rate of change=(stress be after 28 days sucrose partially degree of having a liking for-stress before sucrose degree of having a liking for partially)/stress before sucrose degree of having a liking for * 100% partially
Each group experimental data is processed with the SPSS statistical software, is carried out the t check, investigate neohesperidin group and pathological model group rat sucrose partially degree of having a liking for rate of change whether have significant difference.
5. experimental result
By statistics, each sample sets rat sucrose partially degree of having a liking for rate of change situation see Table 4.
Use SPSS software, carry out the t check, pathological model group and blank group before and after administration sucrose partially degree of having a liking for rate of change relatively have extremely significant difference (P＜0.01), show under the state of chronic stress, rat is not having to have caused its disappearance to the interest of original foundation under pharmaceutical intervention, sucrose degree of having a liking for rate of change partially is starkly lower than blank group, points out simultaneously the chronic stress model to be successfully prepared; Neohesperidin group, positive control drug group and pathological model group before and after the administration sucrose partially degree of having a liking for rate of change more all has difference (P＜0.01) extremely significantly, show that neohesperidin and positive control drug fluoxetine Hydrochloride all can have significant resistant function to the rat body weight anhedonia symptom that causes due to chronic stress, the prompting neohesperidin has opposing and causes the pharmacologically active of the brain malfunction relevant to anhedonia due to depression.
Each sample sets rat sucrose of table 4. chronic stress rat model is degree of having a liking for rate of change partially
Example 5. monoamine oxidase A suppress active testing
1. sample and reagent
Neohesperidin: be purchased from Products in China and examine and determine institute, content 98%
Pargyline: sigma company product, specification 1g/ bottle
Sodium phosphate, sucrose all are purchased from Beijing chemical reagents corporation
MAO active testing test kit: be purchased from Nanjing and build up bio-engineering research institute, test kit forms:
Reagent one: aniline solution (0.016mol/L)
Reagent two: pH7.6Tris buffer
Reagent is crossed solution chlorate's (reaction terminating liquid) at three: 10%
Reagent four: cyclohexane extraction reagent
2. experimental technique
monoamine neurotransmitter is the medium of nervous system conducted signal, its normal presence is the prerequisite that nervous system normally plays a role, in the study of incident mechanism of depression, researcher finds that the minimizing of monoamine neurotransmitter and the morbidity of depression have dependency, and the activity of monoamine oxidase A (MAO-A) increases the monoamine neurotransmitter minimizing that directly causes synaptic space extremely, some antidepressants of clinical practice just are based on monoamine hypothesis design (as imipramine, phenelzine etc.), therefore, seeking oxidase inhibitor is to find to have one of approach of antidepressant activity medicine.
The oxidation under the MAO effect of this experimental basis benzylamine generates the principle of benzyl aldehyde, measures the amount of oxidation product, the activity of indication MAO-A in 242nm wavelength place trap with the cyclohexane extraction extract product.
3. experimentation
(1) preparation of the thick enzyme of rat liver mitochondria monoamine oxidase, MAO (MAO): with Sprague-Dawley rat sacrificed by decapitation, get liver, with 4 ℃ of buffer (pH=7.6, 0.2M sodium phosphate buffer) repeatedly rinse well, take 10g, shred rear grinding, break into homogenate with high speed disperser, the centrifugal 10min of 0.3M sucrose solution 3000rpm that adds pre-cooling with 1: 20 (w/v), get supernatant, the centrifugal 30min of 10000rpm, getting precipitation is dissolved in the 40ml buffer, high speed disperser is beaten even, divide and be filled to 40 pipes (1ml/ pipe), and mensuration protein content (biuret method),-80 ℃ of storages are standby.
(2) preparation of specimen solution: precision takes 50mg neohesperidin reference substance, ultrasonic dissolution in the 10m1 volumetric flask, standardize solution, draw respectively the solution of different volumes from this titer, be mixed with the test solution of a series of variable concentrations, make it add the concentration after enzyme liquid and reagent one and reagent two to be respectively 400,200,100,50,25,6.25,3.125 μ g/ml.
(3) MAO-A suppresses active assay method: add MAO-B type inhibitor 300 μ l pargyline pargyline (500nmol/L) to 30ml rat liver mitochondria enzyme, 37 ℃ of temperature are bathed half an hour, to suppress the activity of MAO-B.Add successively each 0.3ml of a series of testing sample solutions in measuring pipe, reagent one (benzylamine substrate liquid) 0.3ml, reagent two (pH7.6Tris buffer) 3ml, and 0.6ml enzyme liquid, blank tube is with physiologic saline for substitute enzyme liquid, and all the other are identical.Place 3h in 37 ℃ of water-baths, respectively add 10% after taking-up and cross chloric acid 0.3ml, respectively add cyclohexane extraction 3.0ml, rotation mixing 2min on vortex mixer, centrifugation (2000r/min) 10min gets supernatant in quartz colorimetric utensil, returns to zero with blank, measure absorbance in the 242nm wavelength, measure MAO-A active.Take the enzymatic activity of the matched group that only adds the substrate without inhibitor as 100%, calculate determinand to the suppression ratio of MAO-A activity.
MAO-A maximum inhibition=(matched group absorbance-test group absorbance)/matched group absorbance * 100%
4. experimental result
Neohesperidin is to the active IC that suppresses of MAO-A
50Value is 13.14 μ M, with positive control drug moclobemide IC
50Value (5.25 μ M) is close, and result shows, neohesperidin has the effect of significant inhibition MAO-A activity, and it is active that the prompting neohesperidin has the antidepressant pharmacology.
Above-mentioned experiment shows, neohesperidin has antidepressant activity, can comprise with pharmaceutical carrier combination pharmaceutically commonly used being prepared into preparation commonly used, as pharmaceutically acceptable dosage forms such as tablet, capsule, solution, suspension, injection, drip liquid.Can adopt oral, non-intestinal or topical routes.
Claims (3)
1. neohesperidin as the application of antidepressant drug, is characterized in that: the application of neohesperidin in the preparation antidepressant drug.
2. neohesperidin or its pharmaceutically acceptable aglycon or the application of secondary glycosides in the preparation antidepressant drug through hydrolysis or enzymolysis generation.
3. according to claim 1-2 described purposes, it is characterized in that: pharmaceutical dosage form is tablet, capsule, solution, suspension, injection, drip liquid or freeze-dried powder etc.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103341181A (en) * | 2013-06-17 | 2013-10-09 | 中南大学湘雅医院 | Method for detection of effects of neohesperidin for resisting depression and promoting gastrointestinal motility |
| CN104622881A (en) * | 2013-11-15 | 2015-05-20 | 中国药科大学 | Medicine composition and application thereof in preparation of medicines for treating depression |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1416880A (en) * | 2002-11-27 | 2003-05-14 | 北京中医药大学 | New use of powder for regulating liver and spleen and its active part |
| CN1416881A (en) * | 2002-11-27 | 2003-05-14 | 北京中医药大学 | Medicine composition for treating depression and its prepn |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1416880A (en) * | 2002-11-27 | 2003-05-14 | 北京中医药大学 | New use of powder for regulating liver and spleen and its active part |
| CN1416881A (en) * | 2002-11-27 | 2003-05-14 | 北京中医药大学 | Medicine composition for treating depression and its prepn |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103341181A (en) * | 2013-06-17 | 2013-10-09 | 中南大学湘雅医院 | Method for detection of effects of neohesperidin for resisting depression and promoting gastrointestinal motility |
| CN104622881A (en) * | 2013-11-15 | 2015-05-20 | 中国药科大学 | Medicine composition and application thereof in preparation of medicines for treating depression |
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Application publication date: 20130612 |