CN110201118B - Application of traditional Chinese medicine composition in preparation of medicine for treating or preventing vascular senile dementia - Google Patents
Application of traditional Chinese medicine composition in preparation of medicine for treating or preventing vascular senile dementia Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/55—Glands not provided for in groups A61K35/22 - A61K35/545, e.g. thyroids, parathyroids or pineal glands
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/62—Leeches; Worms, e.g. cestodes, tapeworms, nematodes, roundworms, earth worms, ascarids, filarias, hookworms, trichinella or taenia
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/236—Ligusticum (licorice-root)
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/286—Carthamus (distaff thistle)
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/29—Berberidaceae (Barberry family), e.g. barberry, cohosh or mayapple
- A61K36/296—Epimedium
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
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- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
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- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61K2236/30—Extraction of the material
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention relates to the technical field of traditional Chinese medicines, in particular to application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia. The Chinese medicinal composition is prepared from Saviae Miltiorrhizae radix, Notoginseng radix, Carthami flos, herba Epimedii, radix Puerariae, radix Curcumae, semen Persicae, rhizoma Ligustici Chuanxiong, Hirudo, Borneolum, Moschus, and caulis Et folium Ginseng total saponin. The traditional Chinese medicine composition can obviously improve the spatial cognitive memory capacity and explore behaviors of rats with vascular dementia, and the mechanism of the traditional Chinese medicine composition playing a role in neuroprotection is possibly clearly related to the anti-inflammatory and anti-oxidative stress injury functions of the traditional Chinese medicine composition. And also shows obvious advantageous effects compared with the comparison sample prepared by the prior art.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia.
Background
In recent years, the incidence of vascular dementia, the disease of which is increasing year by year as the population of the world is getting older, Vascular Dementia (VD) is the second dementia disease next to Alzheimer's Disease (AD). Not only seriously affects the normal life of the old, but also causes great burden to the family and the society. Therefore, the treatment of Vascular Dementia (VD) is also a hot topic in modern medical field.
At present, a plurality of pharmaceutical compositions for treating vascular dementia are available, such as: the patent publication No. CN109260260A discloses a medicine for treating coronary heart disease and angina pectoris and its preparation method, wherein the medicine comprises 1-3 parts of musk, 400 parts of Ligusticum wallichii 200-. The grant publication No. CN 1679787A discloses a Chinese medicinal preparation for treating vascular diseases, its preparation method and quality control method, wherein the main components of the preparation are capsule, soft capsule or dripping pill of Saviae Miltiorrhizae radix, Notoginseng radix, Carthami flos, herba Epimedii, radix Puerariae, radix Curcumae, Borneolum Syntheticum, Moschus, and total saponins of ginseng stem and leaf. The authorization publication number CN 108853294A discloses a medicinal composition for treating vascular dementia, a preparation method and application thereof, wherein the medicinal composition is prepared from the following raw materials in parts by weight: 10-30 parts of ligusticum wallichii, 5-20 parts of angelica sinensis, 1-15 parts of scutellaria baicalensis, 1-12 parts of rhizoma acori graminei, 1-12 parts of poria cocos, 2-12 parts of polygala tenuifolia, 1-12 parts of ginseng and the like. The formula disclosed above has more medicinal ingredients and complex components, and the quality of the product is difficult to control.
The treatment methods of Western medicines for VD are mainly used for controlling cerebrovascular risk factors, improving cerebral circulation, promoting cerebral metabolism, improving the level of ethylcumine in brain and the like, and although the cognitive and memory functions of a patient are improved to a certain extent, the defects of insignificant curative effect, strong medicine toxicity, multiple side effects and the like exist.
The Moschus XINNAOTONG tablet comprises Saviae Miltiorrhizae radix, Notoginseng radix, Carthami flos, herba Epimedii, radix Puerariae, radix Curcumae, semen Persicae, rhizoma Ligustici Chuanxiong, Hirudo, Borneolum, artificial Moschus, and caulis Et folium Ginseng total saponin. Mainly used for promoting blood circulation, removing blood stasis, inducing resuscitation and relieving pain, and used for treating apoplexy, meridian obstruction, coronary heart disease and angina pectoris caused by blood stasis and collateral obstruction, with the following symptoms: chest oppression, stabbing pain, facial distortion and hemiplegia. At present, no report related to the research of the application of the product to vascular dementia exists.
Disclosure of Invention
In view of the above, the invention provides an application of a Chinese medicinal composition in preparing a medicament for treating or preventing vascular senile dementia. The pharmaceutical composition can obviously improve the spatial cognitive memory capacity and research behaviors of rats with vascular dementia, and the mechanism of the pharmaceutical composition for playing the neuroprotective effect is definitely related to the anti-inflammatory and anti-oxidative stress injury effects of the pharmaceutical composition.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia, wherein the traditional Chinese medicine composition is prepared from salvia miltiorrhiza, pseudo-ginseng, safflower, epimedium, pachyrhizua angulatus, radix curcumae, peach kernel, ligusticum wallichii, leech, borneol, musk and total saponins of ginseng stem and leaf.
The traditional Chinese medicine treatment is carried out based on syndrome differentiation, such as tonifying liver and kidney, reducing phlegm, inducing resuscitation, promoting blood circulation, removing blood stasis and the like. The product can directly act on brain cell tissue to repair and activate damaged and dormant brain cells according to the theory of guiding medicine to human brain and assisting medicine to go upwards in traditional medicine. Therefore, the product has certain action advantages in treating vascular dementia.
Preferably, the traditional Chinese medicine composition is prepared from 10-12 parts by weight of salvia miltiorrhiza, 0.5-1 part by weight of panax notoginseng, 2-4 parts by weight of safflower, 4-6 parts by weight of epimedium, 4-6 parts by weight of pachyrhizua angulatus, 2-4 parts by weight of radix curcumae, 9-12 parts by weight of peach kernel, 1-3 parts by weight of ligusticum wallichii, 1-3 parts by weight of leech, 0.1-0.3 part by weight of borneol, 0.005-0.01 part by weight of musk and 0.05-0.1 part by weight of total saponins of ginseng stem and leaf.
Preferably, the traditional Chinese medicine composition is prepared from 10 parts by weight of salvia miltiorrhiza, 0.5 part by weight of pseudo-ginseng, 2.5 parts by weight of safflower, 4 parts by weight of epimedium, 4 parts by weight of pueraria thomsonii, 2.5 parts by weight of curcuma aromatica, 9.8 parts by weight of peach kernel, 1 part by weight of ligusticum wallichii, 1.3 parts by weight of leech, 0.1 part by weight of borneol, 0.008 part by weight of musk and 0.08 part by weight of total saponins of ginseng stem and leaf.
Preferably, the preparation method of the traditional Chinese medicine composition comprises the following steps:
(1) crushing leeches, adding 8-12 times of water by weight, homogenizing, soaking and filtering; taking filter residues, adding 3-5 times of water by weight, homogenizing, soaking and filtering; mixing filtrates, centrifuging, and collecting supernatant to obtain medicinal liquid A;
(2) pulverizing the salvia miltiorrhiza, the safflower, the peach kernel, the radix puerariae, the ligusticum wallichii, the radix curcumae and the epimedium herb, adding water with the weight of 6-10 times of that of the powder, soaking, carrying out reflux extraction, and filtering the water extract to obtain a liquid medicine B;
(3) mixing the medicinal liquid A and B, concentrating, and drying under reduced pressure to obtain extract A;
(4) pulverizing panax notoginseng, adding an ethanol aqueous solution with the weight of 8-10 times of that of the panax notoginseng, performing reflux extraction, filtering, recovering ethanol, performing reduced pressure concentration, loading the obtained product on macroporous resin, sequentially eluting with water, 30% ethanol and 70% ethanol, collecting 70% ethanol eluent, concentrating, and performing reduced pressure drying to obtain panax notoginseng saponins;
(5) pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin, and mixing.
In the embodiment provided by the invention, the step (1) is as follows: pulverizing Hirudo, adding 8 times of water, homogenizing, soaking, and filtering; taking filter residue, adding 3 times of water by weight, homogenizing, soaking and filtering; mixing filtrates, centrifuging, and collecting supernatant to obtain medicinal liquid A.
Preferably, in the step (1), the soaking temperature is 0-8 ℃, and the soaking time is 10-14 hours; homogenizing for 30-60 minutes; the rotating speed of the centrifugation is 3000-5000 r/min, and the time of the centrifugation is 5-10 minutes.
Preferably, in the step (1), the soaking temperature is 4 ℃, and the soaking time is 12 hours; the homogenization time was 30 minutes; the rotating speed of the centrifugation is 4000r/min, and the time of the centrifugation is 5 minutes.
In the embodiment provided by the invention, in the step (2), the salvia miltiorrhiza, the safflower, the peach kernel, the radix puerariae, the ligusticum wallichii, the radix curcumae and the epimedium are taken, crushed, added with 6 times of water by weight, soaked, extracted by refluxing, and filtered to obtain the liquid medicine B.
Preferably, in the step (2), the soaking time is 0.5-2 hours, the reflux extraction time is 1-3 hours each time, the reflux extraction frequency is 2-3 times, the water extract is filtered, and the filtrates are combined.
Preferably, in step (2), the soaking time is 0.5 hour, the time of each reflux extraction is 1 hour, the number of reflux extractions is 3 times, the water extract is filtered, and the filtrates are combined.
Preferably, in the step (3), the concentration is carried out until the relative density is 1.25 to 1.35 at 50 ℃, and the temperature for decompression drying is 40 to 60 ℃.
Preferably, in the step (3), the concentration is carried out until the relative density is 1.25-1.35 at 50 ℃, and the temperature for decompression drying is 50 ℃.
In the embodiment provided by the invention, in the step (4), the pseudo-ginseng is crushed and added with 8 times of ethanol water solution for reflux extraction.
Preferably, in the step (4), the ethanol aqueous solution has a volume percentage of 70-80%.
Preferably, in the step (4), the ethanol aqueous solution has a volume percentage content of 70%.
Preferably, in the step (4), the reflux extraction is performed for 2 to 3 times, each time for 1 to 3 hours.
Preferably, in step (4), the number of reflux extractions is 2, each for 2 hours.
Preferably, the medicament for treating or preventing vascular senile dementia also comprises pharmaceutically acceptable auxiliary materials.
Preferably, the dosage form of the medicament for treating or preventing vascular senile dementia is granules, tablets, capsules or dripping pills.
The invention provides an application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia, wherein the traditional Chinese medicine composition is prepared from salvia miltiorrhiza, pseudo-ginseng, safflower, epimedium herb, kudzu, turmeric root-tuber, peach kernel, szechuan lovage rhizome, leech, borneol, musk and ginseng stem leaf total saponin. The invention has the technical effects that:
the traditional Chinese medicine composition can obviously improve the spatial cognitive memory capacity and explore behaviors of rats with vascular dementia, and the mechanism of the traditional Chinese medicine composition playing a role in neuroprotection is possibly clearly related to the anti-inflammatory and anti-oxidative stress injury functions of the traditional Chinese medicine composition. And also shows obvious advantageous effects compared with the comparison sample prepared by the prior art.
The invention improves the preparation process of the existing prescription, combines the drug effect, determines the combined preparation method of the invention, improves the taste, reduces the dosage and has obvious advantages in the aspect of treating vascular dementia.
Detailed Description
The invention discloses application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia, and a person skilled in the art can use the contents to refer to the contents and appropriately improve process parameters to realize the purpose. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The formula (weight ratio) of the pharmaceutical composition of the invention is as follows:
10 to 12 portions of salvia miltiorrhiza, 0.5 to 1 portion of panax notoginseng, 2 to 4 portions of safflower, 4 to 6 portions of epimedium, 4 to 6 portions of kudzu, 2 to 4 portions of radix curcumae, 9 to 12 portions of peach kernel, 1 to 3 portions of rhizoma ligustici wallichii, 1 to 3 portions of leech, 0.1 to 0.3 portion of borneol, 0.005 to 0.01 portion of artificial musk and 0.05 to 0.1 portion of total saponins of ginseng stem leaves.
The preparation method of the pharmaceutical composition comprises the following steps:
(1) crushing leeches into coarse powder, adding 8-12 times of water, homogenizing for 30-60 minutes by a high-speed disperser, adding 8-12 times of water for supplementing, standing in a refrigerator, soaking for 12 hours, filtering, reserving filtrate, adding 3-5 times of water, homogenizing for 30-60 minutes by the high-speed disperser, adding 3-5 times of water for supplementing, standing in the refrigerator, soaking for 12 hours, filtering, combining the filtrates, centrifuging at a low speed of 4000r/min, centrifuging for 5-10 minutes, taking supernatant, and reserving in the refrigerator at a temperature of 4 ℃ for later use to obtain liquid medicine A.
(2) Crushing salvia miltiorrhiza, safflower, peach kernel, radix puerariae, ligusticum wallichii, radix curcumae and herba epimedii into coarse powder, soaking the coarse powder in 6-10 times of water for 0.5-2 hours, adding water for reflux extraction for 2-3 times, extracting the water for 1-3 hours each time, filtering the water extract, and combining the filtrates to obtain a liquid medicine B.
(3) Mixing the liquid medicine A and the liquid medicine B, concentrating to relative density of 1.25-1.35 at 50 deg.C, and drying under reduced pressure to obtain extract A.
(4) Pulverizing the panax notoginseng into coarse powder, performing reflux extraction for 2 times by using 70-80% ethanol in an amount which is 8-10 times that of the coarse powder, performing filtration for 2 hours each time, recovering the ethanol, performing vacuum concentration, loading the sample on macroporous resin, sequentially eluting by using water, 30% and 70% ethanol, collecting 70% ethanol eluent, concentrating, and performing vacuum drying to obtain the panax notoginseng saponins.
(5) Pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin powder into fine powder, mixing, adding adjuvant, and making into oral preparation such as granule, tablet, capsule, and dripping pill.
Has the advantages that: the invention improves the preparation process of the existing prescription, combines the drug effect, determines the combined preparation method of the invention, improves the taste, reduces the dosage and has obvious advantages in the aspect of treating vascular dementia. The invention has the advantages that: overcomes the complex compatibility of the traditional Chinese medicines and the obvious toxic and side effects of a large number of traditional Chinese medicines, and also shows obvious advantages compared with the blood sugar reducing sample prepared by the prior art. In order to facilitate understanding of the medicinal value of the invention in the aspect of vascular dementia, the pharmaceutical composition of the invention is shown as follows:
the raw materials or auxiliary materials used in the application of the traditional Chinese medicine composition in preparing the medicine for treating or preventing vascular senile dementia can be purchased from the market.
The invention is further illustrated by the following examples:
example 1
The formulation (weight ratio) of the pharmaceutical composition of this example is as follows:
10 portions of salvia miltiorrhiza, 0.5 portion of panax notoginseng, 2.5 portions of safflower, 4 portions of epimedium, 4 portions of kudzu, 2.5 portions of radix curcumae, 9.8 portions of peach kernel, 1 portion of rhizoma ligustici wallichii, 1.3 portions of leech, 0.1 portion of borneol, 0.008 portion of musk and 0.08 portion of total saponins of ginseng stem and leaf.
The preparation method of the pharmaceutical composition of this example is as follows:
(1) crushing leeches into coarse powder, adding 8 times of water, homogenizing for 30 minutes by a high-speed disperser, adding 8 times of water to supplement the water to 8 times, standing in a refrigerator, soaking for 12 hours, filtering, reserving filtrate for later use, adding 3 times of water, homogenizing for 30 minutes by the high-speed disperser, adding 3 times of water to supplement the water to 3 times, standing in the refrigerator, soaking for 12 hours, filtering, combining filtrates, centrifuging at a low speed of 4000r/min for 5 minutes, taking supernatant, and preserving in the refrigerator at 4 ℃ for later use to obtain liquid medicine A.
(2) Pulverizing Saviae Miltiorrhizae radix, Carthami flos, semen Persicae, radix Puerariae, rhizoma Ligustici Chuanxiong, radix Curcumae, and herba Epimedii into coarse powder, soaking in 6 times of water for 0.5 hr, extracting with water under reflux for 3 times, each for 1 hr, filtering the water extractive solution, and mixing filtrates to obtain medicinal liquid B.
(3) Mixing the medicinal liquid A and B, concentrating to relative density of 1.25-1.35(50 deg.C), and drying at 50 deg.C under reduced pressure to obtain extract A.
(4) Pulverizing Notoginseng radix into coarse powder, extracting with 8 times of 70% ethanol under reflux for 2 times, each for 2 hr, filtering, recovering ethanol, concentrating under reduced pressure, loading onto macroporous resin, sequentially eluting with water, 30% and 70% ethanol, collecting 70% ethanol eluate, concentrating, and drying under reduced pressure to obtain Notoginseng radix total saponin.
(5) Pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin powder into fine powder, mixing, adding 10 parts of sucrose, and making into granule.
Example 2:
the formulation (weight ratio) of the pharmaceutical composition of this example is as follows:
12 portions of salvia miltiorrhiza, 1 portion of pseudo-ginseng, 4 portions of safflower, 6 portions of epimedium, 6 portions of kudzu, 4 portions of radix curcumae, 12 portions of peach kernel, 1 portion of rhizoma ligustici wallichii, 3 portions of leech, 0.3 portion of borneol, 0.01 portion of musk and 0.1 portion of total saponins of ginseng stem leaves.
The preparation method of the pharmaceutical composition of this example is as follows:
(1) crushing leeches into coarse powder, adding 12 times of water, homogenizing for 60 minutes by a high-speed disperser, adding 12 times of water to supplement the water, standing in a refrigerator, soaking for 12 hours, filtering, reserving the filtrate for later use, adding 5 times of water, homogenizing for 60 minutes by the high-speed disperser, adding 5 times of water to supplement the water, standing in the refrigerator, soaking for 12 hours, filtering, combining the filtrates, centrifuging at the low speed of 4000r/min for 10 minutes, taking the supernatant, and preserving in the refrigerator at the temperature of 4 ℃ for later use to obtain the liquid medicine A.
(2) Pulverizing Saviae Miltiorrhizae radix, Carthami flos, semen Persicae, radix Puerariae, rhizoma Ligustici Chuanxiong, radix Curcumae, and herba Epimedii into coarse powder, soaking in 10 times of water for 2 hr, extracting with water under reflux for 2 times, 3 hr each time, filtering the water extractive solution, and mixing filtrates to obtain medicinal liquid B.
(3) Mixing with the medicinal liquid A, concentrating to relative density of 1.25-1.35(50 deg.C), and drying under reduced pressure at 60 deg.C to obtain extract A.
(4) Pulverizing Notoginseng radix into coarse powder, extracting with 10 times of 75% ethanol under reflux for 2 times, each for 2 hr, filtering, recovering ethanol, concentrating under reduced pressure, loading onto macroporous resin, sequentially eluting with water, 30% and 70% ethanol, collecting 70% ethanol eluate, concentrating, and drying under reduced pressure to obtain Notoginseng radix total saponin.
(5) Pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin powder into fine powder, mixing, granulating, adding magnesium stearate, mixing, tabletting, and making into tablet.
Example 3:
the formulation (weight ratio) of the pharmaceutical composition of this example is as follows:
11 parts of salvia miltiorrhiza, 0.8 part of pseudo-ginseng, 3 parts of safflower, 5 parts of epimedium, 5 parts of kudzu, 5 parts of radix curcumae, 12 parts of peach kernel, 2 parts of ligusticum wallichii, 3 parts of leech, 0.3 part of borneol, 0.01 part of musk and 0.08 part of total saponins of ginseng stem and leaf.
The preparation method of the pharmaceutical composition of this example is as follows:
(1) crushing leeches into coarse powder, adding 10 times of water, homogenizing for 45 minutes by a high-speed disperser, adding water to make up to 10 times, standing in a refrigerator, soaking for 12 hours, filtering, reserving filtrate for later use, adding 4 times of water, homogenizing for 45 minutes by the high-speed disperser, adding water to make up to 4 times, standing in the refrigerator, soaking for 12 hours, filtering, combining filtrates, centrifuging at a low speed of 4000r/min for 5 minutes, taking supernatant, and preserving in the refrigerator for later use at 4 ℃ to obtain liquid medicine A.
(2) Pulverizing Saviae Miltiorrhizae radix, Carthami flos, semen Persicae, radix Puerariae, rhizoma Ligustici Chuanxiong, radix Curcumae, and herba Epimedii into coarse powder, soaking in 8 times of water for 1 hr, extracting with water under reflux for 3 times, each for 2 hr, filtering the water extractive solution, and mixing filtrates to obtain medicinal liquid B.
(3) Mixing the medicinal liquid A and B, concentrating to relative density of 1.25-1.35(50 deg.C), and drying at 50 deg.C under reduced pressure to obtain extract A.
(4) Pulverizing Notoginseng radix into coarse powder, extracting with 8 times of 60% ethanol under reflux for 2 times, each for 2 hr, filtering, recovering ethanol, concentrating under reduced pressure, loading onto macroporous resin, sequentially eluting with water, 30% and 70% ethanol, collecting 70% ethanol eluate, concentrating, and drying under reduced pressure to obtain Notoginseng radix total saponin.
(5) Pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin powder into fine powder, adding corresponding adjuvants, mixing, and making into capsule.
Test examples drug efficacy test
Drug effect comparison samples:
the pharmaceutical composition refers to the pharmaceutical composition prepared according to example 1 of the present invention. Comparative sample 1 was prepared according to the grant publication No. CN109260260A specification example 1. Comparative sample 2 was obtained by preparing a sample according to the teaching of the published application No. CN 1679787a, example 2. Comparative sample 3 was prepared according to the teaching of the published application No. CN 108853294 a, example 2.
Instrumental reagent:
SMP500-18272-LSIO microplate reader: manufactured in China Designed in California USA; model LD5-2A low speed centrifuge: beijing medical centrifuge manufacturers; model JA1103N electronic balance: shanghai Minqiao precision scientific instruments, Inc.; morris rat water maze instrument, acetylcholine Ach and total cholinesterase TchE kit.
Experimental animals:
wistar rats, male and female halves, SPF grade, 180-.
The drug composition of the invention and the comparative samples 1, 2 and 3 are used for the comparative study of the drug effect experiment, and the method results are as follows:
1. effect of pharmaceutical composition on vascular dementia rats
1.1 model building
After the rats are purchased, the abnormality is observed for 1 week, swimming adaptability training and Morris water maze experiments are carried out, unqualified rats are removed, and a modified bilateral common carotid artery permanent ligation method (2-VO) is adopted to prepare a vascular dementia rat model.
Weighing a rat, then carrying out intraperitoneal injection (lg/kg) anesthesia by using 10% chloral hydrate, shaving the neck of the rat after the anesthesia takes effect, and disinfecting; a median incision of a rat neck is taken, skin is incised, tissues of all layers are separated in a blunt manner, bilateral common carotid arteries are found out, and double silk threads are ligated to avoid damage to a cervical sympathetic nerve and a vagus nerve. The control group was given the same procedure, but only the common carotid artery was isolated without ligation. The evaluation grading standard of the neurological deficit degree by referring to the zealonga method 4h after the rat is anesthetized and revived after the model building experiment is as follows, grade 0 is carried out, and no defect exists; grade 1, inability to extend contralateral forelimb; grade 2, contralateral forelimb flexion; level 3, slightly turning to the opposite side; grade 4, severe in-place revolutions; grade 5, paralysis of the contralateral side, or even death. And (4) selecting animals successfully molded (animals with the scores of 2 and 3 are selected according to a ZeaLonga grade 5 grading method), and screening 60 successful animal models.
1.2 grouping and administration
120 rats, 10 blank groups, false operation and model building of other rats, 80 survivors are taken and randomly divided into 8 groups: the model group, the comparison sample 1, the comparison sample 2, the comparison sample 3, the high, medium and low dose groups of the pharmaceutical composition are 1.2, 0.6, 0.3 g/(kg. d), and the brain rehabilitation group is 1 g/(kg. d), after operation for 7 days, rats in each group are administrated by gastric gavage, 1 time per day and 10ml/kg are continuously administrated by gastric gavage for 45 days, the model group is administrated with distilled water with the same dose, and the blank group is not treated.
1.3 detection index
1.3.1 behavioural experiments were tested using the Morris water maze: the water maze is a round stainless steel water pool with the diameter of 150cm, the height of 50cm, the water depth of 30cm, the water temperature controlled at (25 seconds 1) ° C, the water pool is divided into 4 quadrants, each quadrant marks a water inlet point, any quadrant is selected, a platform is placed in the center, the height of the platform is 28cm, the diameter of the platform is 15cm, a camera is arranged above the maze and connected with a video recorder and a display, and the swimming track of a rat is automatically recorded for analysis.
1.3.1.1 positional navigation experiment: the rats were placed in the water from the entry point of the two quadrants at the same time every morning for a total of 5 days, and the time it took for them to find the platform in 2 minutes (escape latency) and swim path were recorded. If the rat does not find a platform within 2 minutes, the latency is 120 s. And the rats were placed on the platform for 15s and returned to the cages. This experiment seeks the platform by training the rat to swim. Observing its escape latency and swimming path measures the learning ability of rats.
1.3.1.2 space exploration experiments: after the rats are subjected to a positioning navigation experiment, the platform is removed on the 6 th day, a water entry point is selected and put into water, the swimming track and time of the rats in the pool within 2 minutes are recorded, and the ratio of the swimming time and path of each group of rats in the quadrant of the original platform to the total swimming time and path is analyzed. The experiment observes the changes of space learning and memory of the rat after 5 days of training, and the rat with normal learning and memory can remember the space position of the platform, so that the platform position can be found quickly, after the platform is removed, the rat with normal learning and memory can repeatedly find the platform in the original platform quadrant within 2 minutes, and the rat with poor learning and memory can also find the platform in a maze without purpose. By calculating the ratio of the two, the spatial learning and memory ability of the rat can be further reflected.
1.3.2 index determination
1.3.2.1 rat brain tissue Hippocampus Acetylcholinesterase (ACHE), SOD activity and MDA level detection
After the water maze experiment is finished, 5 rats are randomly selected from each group, 10% chloral hydrate is used for intraperitoneal injection anesthesia, then brain tissues are taken, the hippocampus is separated, homogenate is carried out after cracking, centrifugation is carried out for 10 minutes at 12000 r/min, supernatant is left, and the MDA level and SOD activity detection of the hippocampus of the brain tissues of the rats are detected by adopting an ELISA method.
1.3.2.2 serum detection of IL-6 and TNF-alpha
After the water maze experiment is finished, 5 rats are randomly selected from each group, blood is taken from abdominal cavity after 10% chloral hydrate is used for abdominal cavity injection anesthesia, centrifugation is carried out for 15 minutes at 3000 r/min, serum is taken, and IL-6 and TNF-alpha are detected by adopting an enzyme linked immunosorbent assay.
1.4 results of the experiment
1.4.1 escape latency results in the first 5 days of rats
The model group looked for a significant increase in platform latency (P <0.001) compared to the blank control group. Compared with the model group, the latency of the brain rehabilitation group for searching the platform is obviously shortened in two to five days of training (P <0.05, P < 0.01). The latency of searching for the platform on the third day and the fifth day of the comparison sample 1 is obviously shortened (P is less than 0.05), the latency of searching for the platform on the third day, the fourth day and the fifth day of the comparison sample 2 is obviously shortened (P is less than 0.05), and the latency of searching for the platform on the second day and the fifth day of the comparison sample 3 is obviously shortened (P is less than 0.05). The latency is shortened (P is less than 0.05) in the low-dose group, the third-dose group, the fourth-dose group and the fifth-dose group of the pharmaceutical composition in the training of the second, the third, the fourth and the fifth days, the latency is shortened (P is less than 0.05, P is less than 0.01, P is less than 0.001) in the middle and the high-dose groups of the pharmaceutical composition in the training of the first, the second, the third, the fourth and the fifth days, and compared with the comparative samples 1, 2 and 3, the statistical difference (P is less than 0.05) is generated, so that the obvious action advantage is achieved. The results are shown in Table 1.
TABLE 1 Observation of escape latency results for the first 5 days of rats in each group
Note: comparison with model group#P<0.05，##P<0.01，###P<0.001; comparison with control 1 group<0.01; compare ^ P with control 2 groups<0.05; comparison with control 3 groups□P<0.05。
1.4.2 ratio of time and path of swimming in quadrant of original platform to total time and path of swimming of rat after platform removal
Compared with the blank group, the rats in the model control group have significant difference in the primary quadrant residence time ratio and the distance ratio (P < 0.001); compared with the model group, the brain rehabilitation group, the high and medium dose groups of the pharmaceutical composition have significant difference in the ratio of the residence time and the distance ratio at the primary image limit (P <0.05, P <0.01), and the low dose group of the pharmaceutical composition has shortened residence time at the primary image limit (P < 0.05); compared with the groups 1, 2 and 3, the group with high and medium dosage of the pharmaceutical composition has statistical difference (P is less than 0.05), and has obvious action advantage. The results are shown in Table 2.
TABLE 2 ratio of time and path of swimming in quadrant of original platform to total time and path of swimming for rat after platform removal
Note: comparison with model group##P＜0.01，###P is less than 0.001; comparison with control 1 group<0.05; compare ^ P with control 2 groups<0.05; □ P comparison with control 3 group<0.05。
1.4.3 determination of Acetylcholinesterase (ACHE), SOD activity and MDA level content in rat brain tissue
Compared with the blank group, the acetylcholinesterase content and MDA level of the model control group are obviously increased (P is less than 0.01, P is less than 0.001), and the SOD activity is obviously reduced (P is less than 0.001); compared with the model group, the SOD activity can be improved in each group (P <0.05, P <0.01 and P < 0.001); compared with the control samples 1, 2 and 3, the high-dose group and the medium-dose group of the pharmaceutical composition have obviously reduced acetylcholinesterase content and MDA content and have obvious differences (P <0.05, P <0.01 and P < 0.001). As in table 3.
TABLE 3 determination of acetylcholinesterase content, SOD activity and MDA level in rat brain tissue
Note: comparison with blank control group###P<0.001; comparison with model control group<0.05，**P<0.01，***P<0.001; comparison with control 3 groups□P<0.05。
1.4.4 Effect on IL-6, TNF-alpha content in rat serum
Compared with a blank control group, the IL-6 and TNF-alpha content of the model group is increased (P is less than 0.001); compared with the model group, each group can obviously reduce the content of IL-6 and TNF-alpha (P is less than 0.05, P is less than 0.01, P is less than 0.001), and compared with the groups of the comparison samples 1, 2 and 3, the group with high and medium dosage of the pharmaceutical composition has obvious reduction (P is less than 0.05) of IL-6 and TNF-alpha and has obvious action advantage. The results are shown in Table 4.
TABLE 4 determination of IL-6 and TNF-alpha content in rat serum
Note comparison with model group##P＜0.01，###P is less than 0.001; comparison with control 1 group<0.05; compare ^ P with control 2 groups<0.05; □ P comparison with control 3 group<0.05。
1.5 conclusion
The pharmaceutical composition can obviously improve the spatial cognitive memory capacity and explore behaviors of rats with vascular dementia, and the mechanism of the pharmaceutical composition playing the role of neuroprotection is possibly clearly related to the anti-inflammatory and anti-oxidative stress injury functions of the pharmaceutical composition. And also shows obvious advantageous effects compared with the comparison sample prepared by the prior art.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (9)
1. The application of a traditional Chinese medicine composition in preparing a medicine for treating or preventing vascular senile dementia is characterized in that the traditional Chinese medicine composition is prepared from 10-12 parts by weight of salvia miltiorrhiza, 0.5-1 part by weight of pseudo-ginseng, 2-4 parts by weight of safflower, 4-6 parts by weight of epimedium, 4-6 parts by weight of kudzu vine root, 2-4 parts by weight of radix curcumae, 9-12 parts by weight of peach kernel, 1-3 parts by weight of ligusticum wallichii, 1-3 parts by weight of leech, 0.1-0.3 part by weight of borneol, 0.005-0.01 part by weight of musk and 0.05-0.1 part by weight of total saponins of ginseng stem and leaf.
2. The application of claim 1, wherein the traditional Chinese medicine composition is prepared from 10 parts by weight of salvia miltiorrhiza, 0.5 part by weight of panax notoginseng, 2.5 parts by weight of safflower, 4 parts by weight of epimedium, 4 parts by weight of pachyrhizua angulatus, 2.5 parts by weight of radix curcumae, 9.8 parts by weight of peach kernel, 1 part by weight of ligusticum wallichii, 1.3 parts by weight of leech, 0.1 part by weight of borneol, 0.008 part by weight of musk and 0.08 part by weight of ginseng stem and leaf total saponins.
3. The use of claim 1 or 2, wherein the preparation method of the Chinese medicinal composition comprises the following steps:
(1) crushing leeches, adding 8-12 times of water by weight, homogenizing, soaking and filtering; taking filter residues, adding 3-5 times of water by weight, homogenizing, soaking and filtering; mixing filtrates, centrifuging, and collecting supernatant to obtain medicinal liquid A;
(2) pulverizing the salvia miltiorrhiza, the safflower, the peach kernel, the radix puerariae, the ligusticum wallichii, the radix curcumae and the epimedium herb, adding water with the weight of 6-10 times of that of the powder, soaking, carrying out reflux extraction, and filtering the water extract to obtain a liquid medicine B;
(3) mixing the medicinal liquid A and B, concentrating, and drying under reduced pressure to obtain extract A;
(4) pulverizing panax notoginseng, adding an ethanol aqueous solution with the weight of 8-10 times of that of the panax notoginseng, performing reflux extraction, filtering, recovering ethanol, performing reduced pressure concentration, loading the obtained product on macroporous resin, sequentially eluting with water, 30% ethanol and 70% ethanol, collecting 70% ethanol eluent, concentrating, and performing reduced pressure drying to obtain panax notoginseng saponins;
(5) pulverizing extract A, Notoginseng radix total saponin, Borneolum Syntheticum, Moschus, and caulis Et folium Ginseng total saponin, and mixing.
4. The use according to claim 3, wherein in the step (1), the soaking temperature is 0-8 ℃, and the soaking time is 10-14 hours; homogenizing for 30-60 minutes; the rotating speed of the centrifugation is 3000-5000 r/min, and the time of the centrifugation is 5-10 minutes.
5. The use of claim 3, wherein in the step (2), the soaking time is 0.5-2 hours, the time of each reflux extraction is 1-3 hours, the number of reflux extractions is 2-3 times, the water extract is filtered, and the filtrates are combined.
6. The use according to claim 3, wherein in step (3), the concentration is carried out to a relative density of 1.25 to 1.35 at 50 ℃ and the temperature for drying under reduced pressure is 40 to 60 ℃.
7. The use of claim 3, wherein in step (4), the ethanol aqueous solution has a volume percentage of 70% to 80%; the reflux extraction is carried out for 2-3 times, and each time lasts for 1-3 hours.
8. The use of claim 1, wherein the medicament for treating or preventing vascular senile dementia further comprises pharmaceutically acceptable auxiliary materials.
9. The use of claim 1, wherein the dosage form of the medicament for treating or preventing vascular senile dementia is granules, tablets, capsules or dripping pills.
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Effective date of registration: 20220901 Address after: 130103 no.1369 Shunda Road, Changchun City, Jilin Province Patentee after: JILIN XIUZHENG PHARMACEUTICAL NEW MEDICINE DEVELOPMENT Co.,Ltd. Patentee after: Fusong County Chinese Traditional Medicine Co.,Ltd. Address before: 130012 4-6 / F, amendment building, 1369 Shunda Road, Changchun City, Jilin Province Patentee before: JILIN XIUZHENG PHARMACEUTICAL NEW MEDICINE DEVELOPMENT Co.,Ltd. |