CN106176836B - A kind of pharmaceutical composition, preparation method and application - Google Patents
A kind of pharmaceutical composition, preparation method and application Download PDFInfo
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- CN106176836B CN106176836B CN201610671220.6A CN201610671220A CN106176836B CN 106176836 B CN106176836 B CN 106176836B CN 201610671220 A CN201610671220 A CN 201610671220A CN 106176836 B CN106176836 B CN 106176836B
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- A—HUMAN NECESSITIES
- 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
- A61K36/13—Coniferophyta (gymnosperms)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The present invention relates to a kind of drugs, and in particular to a kind of pharmaceutical composition, described pharmaceutical composition include southerm yew root extract and pharmaceutically acceptable carrier, wherein the weight percent of the southerm yew root extract is 0.1-99.9%.Described pharmaceutical composition source is natural, and preparation process is simple, and low to the toxicity of cell, has stronger immunosuppressive action, has broad application prospects.
Description
Technical field
The present invention relates to pharmaceutical fields, and in particular to a kind of pharmaceutical composition, preparation method and application.
Background technique
In recent years, the correlative study of related natural drug treatment autoimmune disease is the research hotspot of pharmacy circle.From
From the point of view of the related pharmaceutical research for treating autoimmunity disease to natural drug at present, Western medicine of the natural drug than current clinical use
Have apparent advantage, mainly pass through immunosupress, immunological regulation, it is anti-oxidant, induce cell apoptosis and influence nerve and pass
Matter and play treatment autoimmune disease effect.The ImmunopharmacologicaAction Action of natural drug is not independently of the single of other function
Effect, but the unified ergasia being closely associated with nerve, endocrine, Immune Regulative Network.Therefore, both at home and abroad from
Prefered method of the drug for the treatment of autoimmune disease as such drug research is found in nature.
Southerm yew (Taxus chinensis var.mairei) is also known as Bark of Taxus mairei, is Gymnospermae pine
China fir guiding principle Chinese yew mesh taxaceae Chinese yew genus plants, specialty in China, be distributed mainly on the Yangtze river basin, Nanling mountain mountain area with
And mountainous region or the trench of the provinces such as Henan, Shaanxi (Qinling Mountains), Gansu, it is that the plant most fast, distribution is most wide is grown in Taxus.
The dry root of square Chinese yew is extract part on the south southerm yew root extract, and chemical component is numerous, mainly
Contain several constituents such as diterpene-kind compound, steroid compound, lignanoid and flavone compound.Have now been found that south
Japanese yew root extract has significant antitumor and anti-inflammatory activity, but there has been no the reports of immunosuppressive activity.
Drug currently used for immunosuppressive therapy has four major class: chemicals, hormone, fungus metabolite and Chinese medicine,
And chemical pharmacy and fungus metabolite have the sub- selectivity of stronger cell, the side effect of hormone is big, and long-time service can cause
Metabolic disorder induces or aggravates infection, and Chinese medicine is comparatively safe and effective.
Summary of the invention
The purpose of the present invention is to provide a kind of pharmaceutical composition, preparation method and application, described pharmaceutical composition has
The effect of immunosuppressive activity, it is small to the toxic side effect of cell, it is with a wide range of applications.
In order to achieve that object of the invention, the invention adopts the following technical scheme:
In a first aspect, described pharmaceutical composition includes that root of Taxus mentions the present invention provides a kind of pharmaceutical composition
Taking object and pharmaceutically acceptable carrier, wherein the weight percent of the southerm yew root extract is 0.1-99.9%,
Such as can be 0.1%, 0.2%, 0.3%, 0.5%, 0.8%, 1%, 2%, 5%, 8%, 10%, 13%, 15%, 18%,
20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,
92%, 95%, 98%, 99% or 99.9%.
In the present invention, southerm yew root extract chemical component is numerous, the effect for having traditional Chinese medicine compound compatibility to use
Fruit can play one to cell due to containing the non-cell toxicities compounds such as lignanoid, flavonoids in southerm yew root extract
Fixed protective effect, compared to single immunosuppressive activity medicine or single taxol, toxic side effect is small, to cell growth
Inhibiting rate is free from side effects well below the inhibiting rate that immunosuppressive activity drug grows cell.
Preferably, the weight percent of the southerm yew root extract is 1-90%, preferably 20-85%, into one
Step is preferably 40-80%.
Preferably, the southerm yew root extract is any one in water extract, alcohol extracting thing or Chinese medical concrete
Kind or at least two mixture.
Preferably, the pharmaceutically acceptable carrier be mannitol, sorbierite, sorbic acid or sylvite, sodium pyrosulfite,
Sodium hydrogensulfite, sodium thiosulfate, cysteine hydrochloride, thioacetic acid, methionine, vitamin A, vitamin C, vitamin E, dimension
Raw element D, azone, EDETATE SODIUM, Ethylenediaminetetraacetic Acid Calcium Salt, the carbonate of monovalence alkali metal, acetate, phosphate and its aqueous solution, hydrochloric acid,
Acetic acid, sulfuric acid, phosphoric acid, amino acid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextrose
Glycosides, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and its derivates, alginates, gelatin, poly- second
Alkene pyrrolidone, propylene glycol, ethyl alcohol, soil temperature 60-80, Span-80, wax, lanolin, atoleine, hexadecanol, is not eaten glycerol
Sub- esters of gallic acid, agar, triethanolamine, basic amino acid, urea, allantoin, calcium carbonate, calcium bicarbonate, surfactant, poly- second
Glycol, cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talcum powder, calcium stearate, magnesium stearate, vegetable oil, phosphatide,
Polyethylene glycol phosphatide and its derivative, oleic acid and oleate, antioxidant, sodium hydrogensulfite, sodium thiosulfate, vitamin C or
In ionic complexing agent any one or at least two mixture.
Preferably, the vegetable oil is soybean oil, midchain oil, olive oil, tea oil, palm oil, Angelica oil, Seabuckthorn Oil, cowherb
Art oil, cnidium oil, pearl barley oil, safflower oil, Zanthoxylum essential oil, in garlic oil any one or at least two mixture.
Preferably, the phosphatide be egg yolk lecithin, it is soybean lecithin, hydrogenated yolk lecithin, hydrogenated soya phosphatide, artificial
In synthetic phospholipid any one or at least two mixture.
Preferably, the polyethylene glycol phosphatide is polyethylene glycol-cephalin, polyethylene glycol-cholesterol, polyethylene glycol-two-
In fatty glyceride, polyethylene glycol-aliphatic ester, polyethylene glycol-fatty amine or polyethylene glycol-fatty alcohol any one or
At least two mixture.
Preferably, the antioxidant is vitamin E.
Preferably, described pharmaceutical composition be tablet, sugar coated tablet, film coated tablet, enteric coated tablet, hard capsule,
Soft capsule, oral solution, mouth containing agent, granule, electuary, pill, powder, paste, sublimed preparation, suspension, solution, injection,
In suppository, ointment, emplastrum, creme, spray, drops or patch any one or at least two combination.
Second aspect, the present invention provide a kind of preparation method of pharmaceutical composition as described in relation to the first aspect, the south
The extracting method of japanese yew root extract be water mentions, alcohol extracting, ester mentions, ketone mentions, chromatograph, vacuum extraction, supercritical fluid extraction,
Any one in ultrasonic extraction, Enzymatic Extraction, microwave loss mechanisms, charged extraction method or semi-bionic extraction or at least two
Combination, preferably water mentions.
Preferably, the southerm yew root extract extracting method the following steps are included:
Root of Taxus is taken, is added water to cook, is filtered, filtrate decompression concentration, with 95% ethanol precipitation, it is small that refrigeration stands 24
When, filtering takes filtrate recycling ethanol, refrigerates and stands 24 hours, and filtering takes filtrate by resin column, is washed to close colourless, use
75% ethanol elution, collects eluent, and filtering is concentrated to obtain the final product.
Preferably, the root of Taxus and the weight ratio of water are 1:(5-15), for example, can be 1:5,1:6,1:7,
1:8,1:9,1:10,1:11,1:12,1:13,1:14 or 1:15, preferably 1:(9-12).
Preferably, the number of the decoction is 2-5 times, such as can be 2 times, 3 times, 4 times or 5 times, preferably 3 times.
Preferably, the time of the decoction is 2-5h, such as can be 2h, 3h, 4h or 5h, preferably 2h.
Preferably, the every 1ml of the concentrate of the concentration is equivalent to crude drug in whole 1-2g.
Preferably, the volume ratio of ethyl alcohol is 40-70% after the precipitating, for example, can be 40%, 41%, 42%, 43%,
45%, 48%, 50%, 52%, 55%, 58%, 60%, 62%, 65%, 68%, 69% or 70%, preferably 60%.
The third aspect, the present invention provide the pharmaceutical composition of one kind as described in relation to the first aspect and are preparing answering in immune drug
With.
Compared with prior art, the invention has the following advantages:
(1) the southerm yew root extract that the present invention extracts has compared to the extract at other positions of southerm yew
Specific immunosuppressive action, and the drug prepared is low to the toxicity of cell, there was only 11.3% to splenocyte growth inhibition ratio；
(2) pharmaceutical preparation of the invention is simple, and drug effect is significant, reachable to B cell conversion inhibiting rate in experiment in vitro
39.5%, to T cell conversion inhibiting rate up to 44.6%, and in experiment in vivo to mouse ear weight inhibiting rate up to 32.4%, it is right
Thymus index inhibiting rate is up to 22.4%, to index and spleen index inhibiting rate up to 15.2%, it is seen that drug of the present invention has stronger
Immunosuppressive action can be used as immunosuppressor use.
Specific embodiment
The technical scheme of the invention is further explained by means of specific implementation.Those skilled in the art should be bright
, the described embodiments are merely helpful in understanding the present invention, should not be regarded as a specific limitation of the invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The preparation of the pharmaceutical composition of the present invention of embodiment 1
The extracting method of the southerm yew extract the following steps are included:
Root of Taxus is taken, 10 times of waters is added to decoct three times, 2 hours every time, merges decoction liquor, filtering, filtrate decompression
It is concentrated into every 1ml and is equivalent to crude drug in whole 1-2g, with 95% ethanol precipitation to alcohol content up to 60%, refrigeration stands 24 hours, filtering,
Take filtrate recycling ethanol, refrigeration stands 24 hours, filtering, takes filtrate by macroporous resin column, be washed to it is close colourless, with 75% second
Alcohol elution, collects eluent, and filtering is concentrated to obtain the final product.
It will extract after obtained southerm yew root extract is mixed with polyethylene glycol and be fabricated to capsule medicine.
2 Validation in vitro of experimental example
Experiment is divided into blank control group (cell liquid is only added), Con A group, and LPS group is administered A group (only plus test medicine),
It is administered B group (adding Con A and test medicine), is administered C group (adding LPS and test medicine).
(1) it pulverizes, is sieved after taking out mice spleen, rinsed 3 times with RPMI 1640 culture medium.
(2) erythrocyte cracked liquid splitting erythrocyte is used, 1000 turns/min is centrifuged 5min.PBS is resuspended after centrifugation, counts thin
Born of the same parents.
(3) inoculating cell is into 96 porocyte culture plates, and 1 × 106Cells/well.
(4) 100 μ L of culture solution is only added in blank control group, Con A group, LPS group.Test drug is then added in administration A, B, C group
The 100 μ L of RPMI1640 culture solution of object.After 1h Con A group and administration B group be added ConA (final concentration of 5 μ g/mL), LPS group with
C group is administered, LPS (final concentration of 10 μ g/mL) is added, sets 5%CO2, 37 DEG C of culture 72h.
(5) 4h before terminating culture, is added MTT (final concentration of 5mg/mL) to each group, 100 μ L DMSO is added after 4h,
OD value is read at microplate reader 570nm.It is thin to calculate separately splenocyte growth inhibition ratio, B according to relevant control experiment for administration group
Dysuria with lower abdominal colic inhibiting rate, T cell convert inhibiting rate.
Activity Results are as shown in table 1 below.
Table 1
Note: Dex is positive control drug dexamethasone
By result in table as it can be seen that when 50 μ g/mL of administration concentration, drug of the present invention is lower relative to control medicine toxicity, and energy
Obvious to inhibit B cell, T cell conversion, inhibitory effect is suitable with 10 μM of comparison medicine dexamethasone, has apparent immunosupress living
Property, and above-mentioned confirmatory experiment equally is had also been made to Taxus leaf extract, although it is also relatively low to cytotoxicity, to B
The conversion of cell and T cell is substantially without inhibiting effect, it is seen that the drug of the present invention prepared by japanese yew root extract is to mouse
Splenic lymphocyte proliferation and conversion have immunosuppressive activity.
Verifying in 3 body of experimental example
Dinitrofluorobenzene is a kind of haptens, is applied to after skin of abdomen and is combined into comlete antigen with skin protein and makes
Sensitization of skin is reinforced once afterwards for 24 hours, is applied to ear again within sensitization the 5th day, causes delayed allergy.Experiment purpose
It is to observe inhibiting effect of the compound to mouse allergic dermatitis, its immunosuppressive activity of preliminary assessment.
Experimental method: selecting ICR mouse, be randomly divided into Normal group, model control group, administration group, and every group 10.By
Try extract daily administration.Abdomen depilation, is evenly coated in abdomen sensitization for 1%DNFB, and rear same procedure is reinforced primary for 24 hours, causes
Quick 5th day, 1%DNFB is uniformly applied to mouse right ear and is attacked, mouse is put to death afterwards for 24 hours, cuts left and right ear, use diameter
Auricle is made in 8mm blunderbuss, weighing, indicates swelling with left and right auricle weight difference, calculates swelling inhibiting rate (%).
Activity Results are as shown in table 2 below.
Table 2
Note: compared with model group, * p ﹤ 0.05
The results show that drug of the present invention shows apparent immunosuppressive activity under oral 100mg/kg dosage in table.
In conclusion japanese yew root extract of the invention is as a kind of natural drug, it is not only low to the toxicity of cell, no
Inhibit the growth of cell, and there is stronger immunosuppressive action, can be used as immunosuppressor use, is expected to replace amcinonide
Object has broad application prospects.
The Applicant declares that the present invention illustrates the process method of the present invention through the above embodiments, but the present invention not office
It is limited to above-mentioned processing step, that is, does not mean that the present invention must rely on the above process steps to be carried out.Technical field
Technical staff it will be clearly understood that any improvement in the present invention, equivalence replacement and auxiliary element to raw material selected by the present invention
Addition, selection of concrete mode etc., all of which fall within the scope of protection and disclosure of the present invention.
Claims (14)
1. a kind of immune medicinal composition, which is characterized in that the immune medicinal composition includes southerm yew root extract
And pharmaceutically acceptable carrier, wherein the weight percent of the southerm yew root extract is 0.1-99.9%；
The extracting method of the southerm yew root extract the following steps are included:
Root of Taxus is taken, is added water to cook, is filtered, filtrate decompression concentration, with 95% ethanol precipitation, ethyl alcohol after the precipitating
Volume ratio is 60%, and refrigeration stands 24 hours, filtering, takes filtrate recycling ethanol, and refrigeration stands 24 hours, filtering, takes filtrate logical
It crosses resin column, is washed to close colourless, with 75% ethanol elution, collect eluent, filtering is concentrated to obtain the final product.
2. immune medicinal composition according to claim 1, which is characterized in that the weight of the southerm yew root extract
Amount percentage is 1-90%.
3. immune medicinal composition according to claim 1, which is characterized in that the weight of the southerm yew root extract
Amount percentage is 20-85%.
4. immune medicinal composition according to claim 1, which is characterized in that the weight of the southerm yew root extract
Amount percentage is 40-80%.
5. immune medicinal composition according to claim 1, which is characterized in that the immune medicinal composition is sugar coated tablet
Agent, film coated tablet, enteric coated tablet, hard capsule, soft capsule, oral solution, mouth containing agent, granule, pill, powder, pellet
Any one in agent, suspension, solution, injection, suppository, ointment, emplastrum, creme, spray, drops or patch
Or at least two combination.
6. a kind of preparation method of immune medicinal composition according to any one of claims 1 to 5, which is characterized in that described
The extracting method of southerm yew root extract the following steps are included:
Root of Taxus is taken, is added water to cook, is filtered, filtrate decompression concentration, with 95% ethanol precipitation, refrigeration stands 24 hours,
Filtering, takes filtrate recycling ethanol, and refrigeration stands 24 hours, filtering, takes filtrate by resin column, be washed to it is close colourless, with 75%
Ethanol elution, collects eluent, and filtering is concentrated to obtain the final product；The volume ratio of ethyl alcohol is 60% after the precipitating.
7. preparation method according to claim 6, which is characterized in that the root of Taxus and the weight ratio of water are 1:
(5-15)。
8. preparation method according to claim 6, which is characterized in that the root of Taxus and the weight ratio of water are 1:
(9-12)。
9. preparation method according to claim 6, which is characterized in that the number of the decoction is 2-5 times.
10. preparation method according to claim 6, which is characterized in that the number of the decoction is 3 times.
11. preparation method according to claim 6, which is characterized in that the time of the decoction is 2-5h.
12. preparation method according to claim 6, which is characterized in that the time of the decoction is 2h.
13. preparation method according to claim 6, which is characterized in that the every 1ml of the concentrate of the concentration is equivalent to primary
Medicine 1-2g.
14. a kind of immune medicinal composition as described in claim 1 is preparing the application in immune drug.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1348797A (en) * | 2001-10-16 | 2002-05-15 | 邹跃翔 | Healthy taxad composite for treating menalgia |
| CN101244991A (en) * | 2008-03-21 | 2008-08-20 | 昆明自主择业集源生物科技有限公司 | Method for extracting and purifying sequoyitol by using solvent deposition |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1348797A (en) * | 2001-10-16 | 2002-05-15 | 邹跃翔 | Healthy taxad composite for treating menalgia |
| CN101244991A (en) * | 2008-03-21 | 2008-08-20 | 昆明自主择业集源生物科技有限公司 | Method for extracting and purifying sequoyitol by using solvent deposition |
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