[summary of the invention]
Insomnia be divided in pathogenic fire derived from stagnation of liver-QI, expectorant heat disturb, deficiency of both the heart and spleen, hyperactivity of fire caused by deficiency of YIN 4 kinds of pattern of syndrome, the present invention carries out determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs mainly for pathogenic fire derived from stagnation of liver-QI pattern of syndrome, adopt Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae), Semen Ziziphi Spinosae, Poria, the Radix Paeoniae Alba, Rhizoma Chuanxiong, Rhizoma et radix valerianae to be that Raw material processing is prepared into peroral dosage form, obtained medicine has effect of heat clearing away suppressing the hyperactive liver, tranquilizing by nourishing the heart.
For achieving the above object, the present invention is by the following technical solutions:
The invention provides a kind of pharmaceutical composition for the treatment of of insomnia patients, be prepared from by the crude drug of following weight parts:
Semen Ziziphi Spinosae 5 ~ 15, Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) 5 ~ 15, Poria 5 ~ 15, the Radix Paeoniae Alba 5 ~ 15, Rhizoma Chuanxiong 5 ~ 15, Rhizoma et radix valerianae 5 ~ 15.
Pharmaceutical composition provided by the invention, is prepared from by the crude drug of following weight parts:
Semen Ziziphi Spinosae 10 ~ 15, Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) 5 ~ 10, Poria 10 ~ 15, the Radix Paeoniae Alba 10 ~ 15, Rhizoma Chuanxiong 5 ~ 10, Rhizoma et radix valerianae 5 ~ 10;
Be prepared from more excellent by the crude drug of following weight parts:
Semen Ziziphi Spinosae 10, Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) 6, Poria 10, the Radix Paeoniae Alba 10, Rhizoma Chuanxiong 6, Rhizoma et radix valerianae 6.
Pharmaceutical composition provided by the invention can be prepared as any one the existing pharmaceutical dosage form in tablet, capsule, granule, oral liquid.
The invention provides a kind of method preparing the pharmaceutical composition for the treatment of of insomnia patients, comprise the steps:
A) get Semen Ziziphi Spinosae, its total amount 5 ~ 8 times amount 80% ethanol extraction secondary of Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae), extract 1 ~ 2 hour at every turn, filter, medicinal residues are for subsequent use; Merging filtrate, reclaims ethanol to without alcohol taste, is concentrated into the extractum that 60 DEG C are surveyed relative density 1.10 ~ 1.20;
B) medicinal residues after Semen Ziziphi Spinosae, Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae) ethanol extraction and Poria, the Radix Paeoniae Alba, Rhizoma Chuanxiong, Rhizoma et radix valerianae four Chinese medicine material is got, 6 ~ 10 times of water gagings adding its total amount decoct twice jointly, 2 hours first times, second time 1.5 hours, filter after merging twice decocting liquid, it is the extractum of 1.10 ~ 1.20 that filtrate is reduced to 60 DEG C of survey relative densities;
C) merge a), two kinds of extractum of b) step gained, stir evenly, add adjuvant or correctives makes clinical acceptable dosage form.
Pharmaceutical composition provided by the invention is as the application prepared in medicament for treating insomnia.
The crude drug that the present invention adopts:
Semen Ziziphi Spinosae: there is the effects such as tranquilizing soporific, analgesia, convulsion, enhancing immunologic function.Also obvious synergism is had with multiple sedative hypnotic.
Radix Stephaniae Sinicae (Radix Stephaniae Dielsianae): different name, Radix Cissi Kerrii (" conventional Chinese herbal medicine color atlas "), stone qin potato, Stephania cepharatha Hayata (" Guangxi Chinese herbal medicine "), DIWUGUI (Nanchuan " conventional Chinese herbal medicine handbook "), Radix Stephaniae Epigaeae (" mountain of papers Chinese herbal medicine ").Property bitter, cold.Sweet, micro-hardship, temperature.Gui Jing: return liver, stomach warp.Dissipating blood stasis stops blooding, subduing swelling and relieving pain.There is analgesia, hypnosis and sedation.
Rhizoma Chuanxiong, acrid in the mouth, warm in nature, return liver, gallbladder, pericardium channel.There are blood-activating and qi-promoting, wind-expelling pain-stopping effect.
Poria: property slightly sweet flavor is flat.Enter the heart, spleen, lung meridian.Eliminating dampness and diuresis, strengthening the spleen stomach function regulating, mind tranquilizing and the heart calming.Ancients claim Poria to be " when four god medicine ", because its effect widely, regardless of the four seasons, by it and various compatibility of drugs, no matter cold, temperature, wind, wet all diseases, can play its unique effects.
The Radix Paeoniae Alba: be the root of ranunculaceae plant Radix Paeoniae.Nature and flavor picric acid is cool.Enter liver, spleen channel.Nourishing blood to suppress the hyperactive liver, pain relieving in slow, yin fluid astringing receives antiperspirant.
Rhizoma et radix valerianae: be the root and rhizome of Valerianaceae plant Rhizoma et radix valerianae.Different name punching Herba lysimachiae capillipedis, deer grass, Radix Et Rhizoma Nardostachyos.Acrid in the mouth is bitter, warm in nature.Enter the heart, liver two warp.For neurasthenia, spirit is uneasy, has calmness of calming the nerves, wind dispelling spasmolytic, promoting regeneration of the tissue and arresting bleeding, pain relieving.
The drug action of the pharmaceutical composition for the treatment of of insomnia patients of the present invention is further illustrated below by way of pharmacodynamics test.
According to " study of tcm new drug guide ", the pharmacodynamics of compositions of the present invention comprises the following aspects: the pharmaceutical composition observed in various dose situation passes through spontaneous activity in mice and the test of outstanding tail, strychnine, pentetrazole and isoniazid cause mice convulsion test, mice sleep dosage and sub-threshold dose pentobarbital sodium Synergism Testing, the impact of mouse hot-plate analgesic test.
One, sedation research
1. on the impact of spontaneous activity in mice
(1) on the impact (spontaneous activity instrument writing-method) of spontaneous activity in mice
Get the healthy KM kind mice 120 that the number of times of screening activity is in advance close, male and female half and half, by body weight from light to renumbering, table of random numbers method is adopted at random animal to be divided into 6 groups: blank group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group.Each group of mice presses table 1 administration, every day 1 time, continuous 7d, and blank group gives isometric 0.5%CMC-Na solution.Before last administration, water 8h is can't help in fasting, and animal is put into autonomic activities instrument by 30min after last gavage, one, every room, after adapting to 3min, starts to clock, the movable number of times of record 5min, as autonomic activities index, takes statistics learn process with this index.The results are shown in Table 1.Experiment is all carried out every day afternoon 2 ~ 6 time.For reducing experimental error, experimentation takes operation repetitive.
Table 1 is on the impact of mice autonomic activities number of times
Compared with blank group, */* * p<0.05/0.01.
Table 1 result shows: high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all obviously can suppress spontaneous activity in mice (p<0.01 ~ 0.05).
(2) on the impact (walking time and the two forelimb method of act) of spontaneous activity in mice
Getting healthy KM kind mice 120, male and female half and half, by body weight from light to renumbering, adopting table of random numbers method at random animal to be divided into 6 groups: blank group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group.After mice being put into organic plastic box (30cm × 20cm × 15cm) endoadaptation 5min before administration, the number of times that in record 2min, walking time (s) and two forelimb are upwards praised is as normal value, press table 2 dosage ig administration again, continuous 7d, blank group gives isometric 0.5%CMC-Na solution.After last administration 30min, be measured in the same method the Activity value of mice.The results are shown in Table 2.Experiment is all carried out every day afternoon 2 ~ 6 time.For reducing experimental error, experimentation takes operation repetitive.
Table 2 is on the impact of mice autonomic activities number of times
Compared with blank group, */* * p<0.05/0.01.
Table 2 result shows: high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all obviously can suppress spontaneous activity in mice (p<0.01 ~ 0.05).
Two, syngignoscism research
1, on sub-threshold dose pentobarbital sodium induced mice sleep index and the impact of the length of one's sleep
Getting healthy KM kind mice 120, male and female half and half, by body weight from light to renumbering, adopting table of random numbers method at random animal to be divided into 6 groups: blank group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group.Each group of mice is by table 3ig administration, and every day 1 time, continuous 7d, blank group gives isometric 0.5%CMC-Na solution.Before last administration, water 8h is can't help in fasting, 30min after last gavage, each group of mouse peritoneal injection pentobarbital sodium 35mg/kg(determines through preliminary experiment), observe the incubation period of mice sleep (righting reflex loss reaches more than 1min), sleep animal number of elements and sleep time.The results are shown in Table 3.Experiment is all carried out every day afternoon 2 ~ 6 time.For reducing experimental error, experimentation takes operation repetitive.
Table 3 is on pentobarbital sodium sub-threshold dose mice dropping asleep latency sleep index and the impact of the length of one's sleep
Compared with blank group, */* * p<0.05/0.01.
Table 3 result shows: high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all have synergism with pentobarbital sodium, and namely high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all have inhibitory action (p<0.01 ~ 0.05) to nervous system.
2, on the impact of the above threshold dose of sodium pentobarbitone induced mice length of one's sleep
Getting healthy KM kind mice 120, male and female half and half, by body weight from light to renumbering, adopting table of random numbers method at random animal to be divided into 6 groups: blank group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group.Each group of mice is by table 4ig administration, and every day 1 time, continuous 7d, blank group gives isometric 0.5%CMC-Na solution.Before last administration, water 8h is can't help in fasting, 30min after last gavage, each group of mouse peritoneal injection pentobarbital sodium 50mg/kg(determines through preliminary experiment), observe incubation period and the sleep time of mice sleep (righting reflex loss reaches more than 1min), the results are shown in Table 4.Experiment is all carried out every day afternoon 2 ~ 6 time.For reducing experimental error, experimentation takes operation repetitive.
Table 4 is on pentobarbital sodium threshold dose mice dropping asleep latency and the impact of the length of one's sleep
Compared with blank group, */* * p<0.05/0.01.
Table 4 result shows: high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all significantly can shorten the dropping asleep latency of pentobarbital sodium threshold dose mice and extend its length of one's sleep (p<0.01 ~ 0.05), point out medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all to have the effect of collaborative pentobarbital sodium, there is syngignoscism.
3, on the impact of insomnic rats sleep phases
Get healthy male Wistar rat 70, body weight 210 ~ 230g, rat electrode for encephalograms embedding, with pentobarbital sodium intraperitoneal injection of anesthesia 40mg/kg, brain solid positioner is fixed, head cropping with sterilization after, cut off skin of head, cross mouth mosquito forceps is become to clamp corner, abundant exposure skull, peel off periosteum, dry absorbent cotton is cleaned, until surface dry is clean open 1mm cross point place respectively at other on the right side of 1mm before 1mm before coronal suture and lambdoid suture and skull center line, clamping 2 screws with special mosquito forceps hammers in skull, make it to contact with cerebral dura mater, but do not penetrate cerebral dura mater, for cortical electrode, for recording brain electricity, copper wire electrode is inserted respectively to record myoelectricity in the cervical muscle of both sides, 4 electrodes are welded on 4 line mini jacks respectively, be fixed on skull by dental cement, sew up, lumbar injection penicillin sodium, continuous injection three days is postoperative to be placed in special lucite cylinder by rat, single cage is raised, put in screened room, recover two days, to be measured.
To bury the rat of recording electrode, every morning, 9:00 was through gavage 0.5%CMC-Na, and recording the brain electricity (EEG) of the normal rat under non-stress state and myoelectricity (EMG) signal, to trace the time be 9:30 ~ 17:30, amounts to 8h.Morning next day, 9:00, gave the 0.5%CMC-Na of same rat same volume again through gavage, was then placed in by rat in insomnia stimulator, by the electric grid stimulation in rats vola portion 8h bottom cage.Stimulus parameter: electric current 0.5mA, the wide 15ms of ripple, frequency 1Hz.Between stimulation period, EEG and EMG record circuit can interrupt automatically, and electricity irritation can be started automatically by two adjustable timing devices.Like this, often stimulate 30s interval 30min, have a sleepless night to cause rat.The time of tracing is 9:30 ~ 17:30, and stimulate after terminating, analyze the tracer signal of EEG and EMG, the animal all reduced by SWS1 and SWS2 is as experimental rat.3rd day, the insomnic rats screened is divided into 6 groups at random by body weight, model control group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group, by table 5 dosed administration, every day 1 time, continuous 7d, 30min after last administration, rat is placed in insomnia stimulator, at the signal of electricity irritation 8h Nei Yi 16 road physiology signal recorder record EEG and EMG.The time of tracing is 9:30 ~ 17:30, and the brain Electrical change feature of com-parison and analysis rat different times before electricity irritation, after stimulation and after administration, with W, SWS1, SWS2, REMS, TST for observation index, the results are shown in Table 5.In experimentation, environment is ventilation, lucifuge, electromagnetic shielding, adopts self-timing light control system rat living environment to be carried out to the occulting light of 12h every day according to period treatment, experimental temperature 23 ± 2 DEG C, relative humidity 45% ~ 50%.For reducing experimental error, experimentation takes operation repetitive.
4, fenclonine (PCPA) causes the impact of rat insomnia model
Get healthy male Wistar rat 70, body weight 180 ~ 220g, by body weight from light to renumbering, table of random numbers method is adopted at random animal to be divided into 7 groups: blank group, model control group, the high, medium and low dosage group of medicine group of the present invention, mind-easing tonic bolus with arborvitate seed group, stable group.Except blank group, all the other respectively organize rats by intraperitoneal injection PCPA300mg/kg, continuous 2 days, copy insomnia animal model, blank group lumbar injection same volume normal saline.Within 3rd day, respectively group rat is by table 6ig administration, and every day 1 time, continuous 7d, blank group gives isometric 0.5%CMC-Na solution.Before last administration, water 8h is can't help in fasting, 60min after last gavage, by each group of rat sacrificed by decapitation, take out full brain, weigh after decerebellation, get right half side cerebral tissue be placed in 10% neutral buffered formalin solution fix, through the saturating wax of routine, embedding, section, prepare tissue morphology section, immunohistochemical staining measures the expression of cholecystokinin (CCK-8); All the other left half side cerebral tissue add the distilled water of pre-cooling in the ratio of every gram of 1:3, homogenate 6 ~ 8min, 12000r/min(-4 DEG C), centrifugal 10min, accurate Aspirate supernatant 0.5ml, add acetonitrile 0.5ml, whirlpool mixes 1 minute, in the centrifugal 10min of 10000r/min, draw 15 μ l injection liquid matter combined instruments to analyze, chromatographic condition: mobile phase: methanol-water (50:50), flow velocity: 0.5ml/min, C184.6 × 150mm, 5um; 5-HT: cation MRM pattern, mz177/115 impact energy 30eV; GABA: cation MIM pattern, mz104.2, impact energy 30eV.The results are shown in Table 6.For reducing experimental error, experimentation takes operation repetitive.
Table 6 fenclonine (PCPA) is had a sleepless night the impact that 5-HT, GABA level and CCK-8 express in rat model cerebral tissue
Compared with blank group, */* * p<0.05/0.01.
Table 6 result shows: high, the middle dosage of medicine group of the present invention and Chinese medicine contrast medicine mind-easing tonic bolus with arborvitate seed all significantly can increase the content (p<0.01 ~ 0.05) of 5-HT and GABA in PCPA induced insomnia rat model cerebral tissue, also significantly can increase the expression (p<0.01 ~ 0.05) of CCK-8.
The pharmaceutical composition that test adopts embodiment 6 to prepare.