CN108586628A - A kind of preparation method and applications with the yew amylose for dropping high lithemia effect - Google Patents
A kind of preparation method and applications with the yew amylose for dropping high lithemia effect Download PDFInfo
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Abstract
The invention discloses a kind of preparation method and application for the yew amylose for having and dropping high lithemia effect, the preparation method of specific yew amylose includes the following steps：A it) pre-processes；B) water extract-alcohol precipitation；C it) washs；D) deproteinized；E it) dialyses；F it) is freeze-dried.And the application of the yew amylose that is prepared using this method in anti-trioxypurine is disclosed, and discloses the medicament for the anti-trioxypurine being prepared using the yew amylose that this method is prepared.Present invention extraction yew amylose is at low cost, and recovery rate and purity are high, and obtained yew amylose has effects that reduce high lithemia, has protective effect to kidney.
Description
Technical field
The invention belongs to pharmaceutical technology field more particularly to a kind of preparation sides of the yew amylose with anti-trioxypurine effect
Method and application.
Background technology
Hyperuricemia refers to a kind of fuselage state of the uric acid concentration in blood beyond normal range (NR), it is considered that male
When uric acid concentration in blood is more than 400 μm of ol/L, the uric acid concentration in women blood is considered as when being more than 360 μm of ol/L as height
Uricacidemia.In recent years studies have shown that hyperuricemia is not only the most important biochemical basis of gout, and be also to cause high blood
Pressure, hyperlipidemia, atherosclerosis, obesity, insulin resistance, metabolic syndrome angiocardiopathy occur and death it is important
Factor, it has also become threaten one of main metabolic disease of human health.
The chemicals of clinical treatment hyperuricemia at present, mainly to inhibit the Allopurinol and uricosuric of uric acid generation
Based on the drugs such as eccritic probenecid, Benzbromarone.These adverse drug reactions are more, and patient tolerability is poor, significantly impact height
The treatment of uricacidemia.In recent years, traditional Chinese medicine achieved very big effect in terms of reducing serum uric acid level.
China Patent Publication No. is that the patent of invention of CN101879264A discloses a kind of " Chinese traditional medicine composition for anti-trioxypurine
Object " extracts various composition as raw material using smilax, rhizoma alismatis, is prepared into oral preparation, injection, is used for anti-trioxypurine；Xiao Fuzhou
" medicinal herb tea for reducing uric acid of patient with gout ", using desmodium, Asiatic plantain, Radix Angelicae Sinensis and Ilex Latifolia Thunb as raw material, is crushed into powder, medicine is made
Tea, patient drink tea daily, can anti-trioxypurine；Yi Lian " Chinese medicine preparation and composition of prevention and treatment hyperuricemia and its preparation
Method " forms or extracts alkaloid, volatile oil and flavones therein etc. with " Cortex Phellodendri, rhizoma atractylodis, smilax, caulis lonicerae " proportioning
It matches, finds that there is notable anti-trioxypurine activity；From wherein it will be seen that smilax, rhizoma alismatis are edema-alleviating diuretic；Vehicle
Preceding grass is stranguria-relieving diuretic；Desmodium is damp-excreting anti-icteric medicinal；These can be classified as damp-clearing drug.It can be seen that being controlled in terms of Chinese medicine
It is based on clearing damp and promoting diuresis to treat high lithemia.
Chinese yew has the function of clearing damp and promoting diuresis as medicinal material, and Chinese medicine treats high lithemia based on decocting, through research
It was found that Chinese yew, in addition to it can extract anti-cancer medicine paclitaxel, the main component of water decoction is yew amylose, therefore of the invention
Just based on this point, efficiently using the branches and leaves of yew after industrially extracted complete taxol and 10-DABIII is
Raw material extracts yew amylose, and does purification process, not only economizes on resources, but also further excavates the medicinal of yew amylose
Value is mankind's medical and health field to realize the reasonable of rare Chinese yew resource and maximized effectively application
The exploitation of resource provides more possibilities.
Invention content
The object of the present invention is to provide a kind of yew amylose and preparation method thereof, realize that it is preparing treatment antihyperuricemic
Application in disease drug.
A kind of preparation method of yew amylose provided by the invention comprising following steps：
A it) pre-processes
The dry waste material of Chinese yew after extracted complete taxol and 10-DABIII is taken, is crushed, ethyl alcohol is added, carries out Soxhlet
Extraction, it is spare to be collected by filtration the dregs of a decoction.
B) water extract-alcohol precipitation
Step A) described in the dregs of a decoction appropriate purified water is added, stirring to the dregs of a decoction is waterlogged, and Soxhlet extraction filters, decompression
Concentration；Ethyl alcohol, stand at low temperature is added in concentrate, and filtration obtains extract.
C it) washs
Step B) in it is extract obtained be slowly added dropwise absolute ethyl alcohol, acetone, ethyl acetate successively, it is dry after crude product.
D) deproteinized
Step C) in gained crude product be dissolved in appropriate hot water, with Sevage methods be added 1 times of volume n-butanol/petroleum ether
Mixed liquor stirs, and centrifugation removes a layer aqueous, obtains the polysaccharide solution after removing protein.
E it) dialyses
Step D) in gained polysaccharide solution, using flowing water dialysis remove solution small molecular substance.
F it) is freeze-dried
Step E) in solution decompression concentration after dialysis, be freeze-dried after precooling, obtain Chinese yew Thick many candies.
Preferably, the step A) in be added 80% ethyl alcohol Soxhlet extraction solid-liquid ratio be 1:5-15.
This case is before water extract-alcohol precipitation first with 80% alcohol pre-treatment Chinese yew powder, in order to remove remaining in medicinal material
Fat-soluble compound and part monosaccharide, can improve step B polysaccharide from medicinal materials extraction efficiencies in this way, increase recovery rate and carry
The purity of high red bean polysaccharide.The solid-liquid ratio of ethyl alcohol Soxhlet extraction influences whether the degree of removal of impurities, and the present invention is by repeatedly attempting, really
The solid-liquid ratio of fixed 80% ethyl alcohol Soxhlet extraction is 1:With good dust removal rate when 5-15, and cost is minimum, further preferably
80% ethyl alcohol Soxhlet extraction solid-liquid ratio be 1:8-12, further preferred 1:10.
Step B) water extraction twice is carried out, it is 1 that purified water solid-liquid ratio is added for the first time:8-12, second of addition purified water
Solid-liquid ratio is 1:5-10, step B) in be concentrated under reduced pressure into a concentration of 0.8-15g/mL of crude drug；3-6 times of volume second is added in concentrate
The temperature of alcohol, concentration of alcohol 85-97%, stand at low temperature is 2-8 DEG C, time of repose 15-36h.
Water extract-alcohol precipitation is that current polysaccharide from medicinal materials extracts most common method, and method is similar, but is lacked to extraction process
There are larger differences, and production efficiency is improved in the case where keeping recovery rate, reduces cost, and the present invention carries out water twice and extracts,
It is 1 that purified water solid-liquid ratio is added for the first time:8-12, it is 1 that second, which is added purified water solid-liquid ratio,:5-10, further preferred the
Solid-liquid ratio is 1:10, second is 1:8.
In order to ensure that washing step can effectively remove polysaccharide loss when impurity reduces washing, inventor is washed by observing
The method that polyoses content is tested in journey cleaning solution color change and cleaning solution confirms washers to investigate best washing methods
Skill.
Finally determining washing methods is absolute ethyl alcohol, acetone, ethyl acetate to be added with this, and the volume ensured is all
Southerm yew feeds intake 0.05-0.2 times, preferably 0.1 times of quality.
Step E) in the dialysis bag retention molecular weight 8000-14000 that uses of dialysis.
The present invention also provides a kind of yew amyloses to prepare the application in treating antihyperuricemic disease drug.
Yew amylose in preparing antihyperuricemic disease drug, can be made and pharmaceutically be connect as main active
The various preparations received, the dosage form of preparation include but not limited to：Enema；Tablet；Capsule, such as hard capsule；Oral solution；Mouthful
Containing agent；Granule；Electuary；Pill；Pill；Powder；Paste；Sublimed preparation；Suspension；Pulvis；Solution；Suppository；Liniment；Frost
Agent；Spray；Powder spray；Aerosol；Drops；Pastille and patch.
The auxiliary material for preparing various dosage forms is the excipient substance of pharmaceutical field routine, and auxiliary material includes but not limited to：Adhesive is filled out
Fill agent, diluent, tablet agent, lubricant, disintegrant, colorant, flavoring agent, wetting agent and surfactant etc..
In above-mentioned application, it will be understood by those skilled in the art that can be produced by the conventional production process of pharmaceutical field, red bean
The active component content of China fir polysaccharide is in 1%-99.5%.
The present invention usage amount can according to administration route, the variations such as the weight of patient, the severity of hyperuricemia,
Its dose can be 0.01-2g/kg, can be in single or divided doses.
The present invention is using the dry waste material of Chinese yew after extracted complete taxol and 10-DABIII as extraction Chinese yew
The raw material of polysaccharide reduces cost, while increasing the utilization rate of Chinese yew, and it is more by the setting of technique to improve Chinese yew
The recovery rate and purity of sugar, while it is non-toxic to demonstrate the yew amylose that extraction obtains.
The effect of present invention finds yew amyloses to reducing high lithemia, and corresponding preparation has been prepared, and verify
With yew amylose be active constituent made of drug in the experiment of high lithemia mouse species, show that yew amylose is excellent
Anti-trioxypurine and its depressor muscle acid anhydride effect, and have protective effect to kidney；It is that active constituent is made drug and exists with yew amylose
Yew amylose is showed in clinical trial apparent anti-trioxypurine effect.
Specific implementation mode
To be best understood from the present invention, further specific elaboration is made to the present invention below by following embodiment, but not
It can be regarded as limitation of the invention.
1 southerm yew total starches preparation process preprocess method of embodiment is explored
Inventor is before water extract-alcohol precipitation first with 80% alcohol pre-treatment Chinese yew powder, in order to remove residual in medicinal material
The fat-soluble compound and part monosaccharide deposited can improve step B polysaccharide from medicinal materials extraction efficiencies in this way, increase recovery rate.Hair
A person of good sense compares four kinds of solid-liquid ratios 1 respectively:5、1:10、1:15、1:20, it is found that solid-liquid ratio is 1:When 5, removal of impurities is incomplete, and feed liquid
Than being 1:15 and 1:Although 20 effects are very good, production cost can be improved, is 1 it is advantageous to solid-liquid ratio:10.
2 southerm yew total starches preparation process water extract-alcohol precipitation Research on Methods of embodiment
Water extract-alcohol precipitation is that current polysaccharide from medicinal materials extracts most common method, and method is similar, but but to extraction process
Verification experimental verification is needed, production efficiency is improved in the case where keeping recovery rate, reduces cost.Inventor uses solid-liquid ratio 1：5、1:
8、1:10、1:15 and extraction 1,2,3 time, using extract recovery rate as reference standard, extraction process is explored, confirm most
Good extraction process：Twice, first time solid-liquid ratio is 1 for extraction:10, second is 1:8.
3 southerm yew total starches preparation process washing methods of embodiment is explored
In order to ensure that washing step can effectively remove polysaccharide loss when impurity reduces washing, inventor is washed by observing
The method that polyoses content is tested in journey cleaning solution color change and cleaning solution, to investigate best washing methods, it is thus identified that washing
Technique：Be added absolute ethyl alcohol, acetone, ethyl acetate volume be all 0.1 times (mass/volume) of southerm yew inventory.
The selection of 4 southerm yew total starches preparation process bag filter of embodiment
Bag filter aperture determines that polysaccharide molecular weight, polysaccharide molecular weight determine its drug effect.Inventor is consulting a large amount of texts
It offers, and by the animal experiment of different molecular weight yew amylose, it is found that when dialysis bag retention molecular weight is 8000-14000,
Drug effect and recovery rate are most suitable.
It is prepared by 5 southerm yew polysaccharide of embodiment
Southerm yew powder 1kg is taken, adds 80% ethyl alcohol 10L, after impregnating 1h, 80 DEG C of Soxhlet extractions 2 hours are collected by filtration
The dregs of a decoction.10L purified waters are first added in the dregs of a decoction, and stirring to the dregs of a decoction is waterlogged, 80 DEG C of Soxhlet extraction 2h, filtration, and residue adds 8L water again
It repeats to extract once, merges extracting solution twice, filtering is concentrated under reduced pressure into a concentration of 1g/mL of crude drug；4 times of volumes are added in concentrate
95% ethyl alcohol, 4 DEG C stand 24 hours, filtration obtain extract.It is extract obtained that the anhydrous second of 100mL is slowly added dropwise successively successively
Alcohol, acetone, ethyl acetate obtain yew amylose crude product after dry.Gained crude product is dissolved in appropriate hot water, is added with Sevage methods
Enter n-butanol/petroleum ether (volume ratio 1 of 1 times of volume:4) mixed liquor, stirring centrifuge after 30 minutes, remove a layer aqueous；It repeats
Aforesaid operations 3 times, obtain the polysaccharide solution after removing protein.It is 8000-14000's that gained polysaccharide solution, which is packed into molecular cut off,
In dialysis band, using 72 hours removal solution small molecular substances of flowing water dialysis.Solution decompression after dialysis is concentrated into crude drug
Concentration 3.0g/mL is freeze-dried after precooling, obtains Chinese yew Thick many candies.
6 southerm yew Thick many candies safety testing of embodiment
A) acute toxinology experiment of the present invention
5 Thick many candies of embodiment used herein.When experiment with distilled water be configured to required concentration by test solution.
The present invention selects SPF grades of kunming mices, each 20 of male and female, 18~22g.It is tested using maximal tolerance dose (MTD) method
The disposable gavage of object is given, poisoning dosage 15.0gkgd-1, 0.8~1.0ml of gavage capacity (20g)-1.It is dynamic before experiment
Object fasting (can't help water) 16h, records the poisoning manifestations and death condition of animal, 14d is observed continuously.
Result of the present invention is as described below：After gavage gives tested material, apparent poisoning symptom is had no, animal is without death in 14d.
The acute toxicity MTD ＞ 15.0gkg of 2 kinds of gender mouse of the tested material pair-1.According to acute toxicity grading criteria, the sample category
Practical nontoxic grade.
B) Genetic toxicity of the present invention
Salmonella reversion test
Using conventional panel incorporation methods, given the test agent Chinese yew Thick many candies are configured to 8,40,200,1000,5000ug/
Ware is tested.After the sterile suction method degerming of solution, be added in top agar respectively 0.1mL test strains enrichment liquids,
0.1mL tested materials solution and 0.5mL S9 mixed liquors (when needing metabolism activation), are poured into after mixing on bottom culture medium flat plate.Together
When be untreated control, solvent control and positive control (TA97a[-S9]:500ug/ ware atabrine；TA98[-S9]:6.0ug/ ware
Daunomycins；TA100[-S9] and TA102[-S9]:1.0uL/ ware methyl methylsulfonates；TA97a[+S9♂]、TA98[+S9] and TA100[+
S9]：10.0ug/ ware 2- aminofluorenes；TA102[+S9]：50.0ug/ ware 1,8- dihydroxy anthraquinones).
As a result as shown in Table 1 and Table 2：Under each concentration of this experiment, regardless of whether S9 mixed liquors are added, return and become clump count
It is not more than beam back parameter certainly 2 times, and has no dose-response relationship.And each positive controls show strong mutagenesis
Effect.According to Salmonella reversion test result criterion, the tested material of this experimental test has no mutagenesis.Illustrate to have no this
Tested material has genetoxic to mouse sperm.
Table 1 first time Salmonella reversion test result (x ± s, n=3)
Note：Positive control：It is not added with and uses S9：TA97a500.0ug/ ware atabrine；TA986.0ug/ ware daunomycins；TA100
And TA1021.0ul/ ware methyl methylsulfonates；Add and uses S9：TA97a, TA98, TA10010.0ug/ ware 2- aminofluorenes；
TA10250.0ug/ ware 1.8- dihydroxy anthraquinones.
The 2nd Salmonella reversion test result (x ± s, n=3) of table 2
Micronucleus test
Each 50 of ♀ ♂ mouse, 25~30g.Sample gavage is given.5 groups are randomly divided by weight, negative control group is (pure
Change water), given the test agent Chinese yew Thick many candies 600,1200,2400mgkg-1Group, positive control (cyclophosphamide, 40mgkg-1) group, each 5 of every group of ♀ ♂.With 0.4mL (20g)-12 gavages (interval is for 24 hours), 6h takes breastbone film-making mirror after the 2nd gavage
Inspection.
As a result：For test group animal under each test dose, polychromatic erythrocyte (PCE) Rate of Appearance Micronuclei, PCE/ maturations are red
Cell (RBC) compared with negative control group, difference there are no significant (P>0.05).And positive controls and negative control group phase
Than the significant (P of difference<0.05) strong mutagenesis, is shown.Test result shows the tested material in this experiment agent
Mutagenesis is had no under amount.
Mouse inbred strain
Each 25 of ♂ mouse, 25~30g.Sample gavage is given.5 groups are randomly divided by weight, negative control group (purifying
Water), given the test agent Chinese yew Thick many candies 600,1200,2400mgkg-1Group, positive control (cyclophosphamide, 40mgkg-1)
Group, every group 5.With 0.4mL (20g)-1Weight gavage, continuous 5d take bilateral epididymal film-making in the 30th day after gavage for the first time
Microscopy.
As a result：The rate of teratosperm of negative control group is 1.99%, the teratospermia of positive controls (cyclophosphamide group)
Rate is 5.26%, compared with negative control group, the significant (P of difference<0.05).And test the rate of teratosperm of 3 dosage groups
Respectively 2.06%, 2.02%, 2.12%, with negative control group than no significant difference (P>0.05).
7 observation experiment of embodiment
A) grouping and dosage setting：It weans SD rats 120,4 groups is randomly divided by weight, every group 30, ♀ ♂ each 15
Only.Tested material Chinese yew Thick many candies are given through feed incorporation.If Normal group：Give full nutrition mixed feed；Test sample
Product are low, in, high 3 dosage groups：600,1200,2400mgkg-1·d-1.Animal single cage is raised, water of freely ingesting, continuously
Observation 30d terminates to feed.
B) observation index
General indicator：Observation animal is primary daily, record animal hair color, behavior and death condition；Respectively at the of experiment
It weighs within 7,15,22,30 days primary, observes weight situation of growth；Weekly plus feed 2 times, record of weighing, and the feeding for spreading mistake is recorded
Doses calculates food consumption and utilization rate.
Hematological examination：In experiment the 30th day, femoral artery took blood to carry out hemoglobinometry, and red blood cell, leucocyte, blood are small
Plate counts and leukocyte differential count measures；Detach serum, using Hitachi -7150 automatic clinical chemistry analyzer carry out total protein (TP),
Albumin (ALB), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST), glucose (GLU), creatinine (CR), urea nitrogen (BUN),
Cholesterol (T-CHE) and triacylglycerol (TG) measure.
As a result：It is the hemoglobin of surveyed each group animal, red blood cell, leucocyte (including classification), total serum protein (TP), white
Albumen (ALB), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST), glucose (GLU), creatinine (CR), urea nitrogen (BUN), courage
Sterol (T-che) and triacylglycerol (TG) are in range of normal value.
C) Organ weight and Histopathology inspection
Off-test, rat use femoral artery sacrificed by exsanguination, and dissection immediately carries out gross examination；Weigh liver, spleen, Bilateral Renal
And testis, calculate separately dirty body ratio；Liver,kidney,spleen, Stomach duodenum, ovary, testis are fixed with formalin.Paraffin embedding is cut
Piece, HE dyeing carry out histopathologic examination.
As a result：Dirty body ratio：The internal organs absolute mass and its liver quality/weight, spleen matter of each dosage group of Chinese yew Thick many candies
Amount/weight, kidney quality/weight, testis quality/body mass ratio there are no significant difference.
Tissue pathology checking：When gross anatomy, control group and each dosage group rat liver,kidney,spleen of yew amylose, stomach, ten
Two duodenum 12, ovary, testis observe no abnormality seen in body, therefore high dose group are only selected to carry out histopathological examination.It is checked under mirror
Except control group and high dose group have individual mile abnormalities (fatty degeneration of liver, renal interstitial stove cell infiltration, stomach and duodenum
Mucosal epithelial cells partial exfoliation simultaneously has outside a small amount of cell infiltration, remaining each example rat tissue pathological examination result is just
In normal range, high dose group relatively has no apparent pathology difference with blank control group, prompt yew amylose to liver,kidney,spleen,
Stomach duodenum, ovary, testis are without apparent damaging action.
The configuration of 8 high lithemia mouse model modeling liquid of embodiment and model construction
A) modeling liquid making method：Weigh 97% Oteracil Potassium (1gkg-1·d-1) and adenine (100mgkg-1·
d-1), respectively 18.0g and 1.8g, mixing are dissolved in 225ml distilled water, you can.
B) model building method：By mouse according to weight, gender, ensure every group of average mice body weight, each semi-uniform of gender,
It is grouped, respectively Normal group (C), model group (M), yew amylose high dose (H-600), the high agent of yew amylose
Measure (H-300), yew amylose high dose (H-150), Allopurinol positive drug group (Y).It is grouped concrete condition such as following table：Such as table 3
It is shown：
3 animal packet situation table of table
In addition to Normal group, every mouse of remaining each group is administered according to 0.25ml/20g, continues 5 days.
C) yew amylose high dose (H-600), yew amylose high dose (H-300), yew amylose high dose (H-
150) test medicine preparation method：Southerm yew total starches 1.5g, 0.75g, 0.375g are weighed respectively, are respectively dissolved in 25ml steamings
In distilled water, it is configured to the test solution of 600mg/kg, 300mg/kg, 150mg/kg concentration, it is more to be respectively designated as Chinese yew
Sugared high dose (H-600), yew amylose high dose (H-300), yew amylose high dose (H-150).
D) allopurinol positive drug is prepared:Allopurinol pharmaceutical specifications are 0.1g/ pieces, take 2, are dissolved in 25.00ml steamings
In distilled water, you can.
E it) is grouped situation table according to step B tables 3, modeling is administered the 6th day, after giving modeling liquid 1h, is given according to 0.2ml/20g
Give medicine Chinese yew Thick many candies height, middle low dosage and Allopurinol positive drug, continued administration 7d.After 7d is administered,
Am12:00 starts to pluck eyeball and takes blood, centrifuges, takes supernatant, be transferred to 5mlEP pipes, stores -20 DEG C of refrigerators and preserve, to be measured.
F) Thick many candies of the present invention reduce the observation of high lithemia mouse
In view of from, compared with normal group, treatment group's animal state is relatively good, and model group animal activity is relatively
Difference；According to H-600, H-300, H-150, Y, animal appearance coat gloss, weight, behavioral activity, food ration, water intake is successively
It reduces.
Compared with normal group, H-600 groups, left and right sides kidney is without atrophy phenomenon, and double kidney colors are slightly whitened, but it is whole with just
Normal group is no different, the uneven unobvious of surface particles.Y positive drug groups, nephrarctia most serious, wherein the most with left side nephrarctia
Seriously, double kidney colors are obviously whitened, and surface particles are uneven apparent.Therefore speculate positive drug Allopurinol in the same of anti-trioxypurine
When, because take for a long time, drug accumulation and there are serious injury of kidney.I.e.：H-600 groups not will produce kidney injury, and to kidney function
There can be protective effect.
G) UA, BUN, Cr result difference are as shown in table 4 in serum：
UA, BUN, Cr test result (X ± S) in 4 mice serum of table
Note：" * " is compared with normal group, and P ＜ 0.05, there are significant differences for model group；" # " is the P compared with model group
＜ 0.05, there are significant differences for test medicine group.
From table 4 it can be shown that model group is compared with normal group, uric acid index, urea nitrogen index and creatinine index are deposited
At significant difference (P ＜ 0.05), and it is apparently higher than the level of normal group mouse, shows mouse hyperuricemia modeling success.
Shown in 4 Serum Indexes of table, uric acid (UA) index, test medicine each group is compared with model group, positive drug Allopurinol group
(Y) and the significant difference of yew amylose H-600 groups (P ＜ 0.05), uric acid index is substantially reduced.And yew amylose H-
The uric acid level that two groups of 300 and H-150 is declined compared with model group, but does not have significant difference.
Shown in urea nitrogen (BUN) index, compared with model group, positive drug Allopurinol group (Y) has significantly test medicine each group
Sex differernce (P ＜ 0.05), positive drug group urea nitrogen levels conspicuousness reduces, and test medicine yew amylose H-600, H-300
There is drop urea nitrogen effect with H-150, but does not have conspicuousness (P>0.05).
Shown in creatinine (Cr) index, for test medicine each group compared with model group, positive drug Allopurinol group (Y), Chinese yew are more
The sugared significant difference of H-600 groups (P ＜ 0.05), creatinine (Cr) conspicuousness reduce, and yew amylose H-300 and H-150 according to
It is secondary to have downward trend, but do not have significant difference (P>0.05).
The conclusion that the present invention obtains：Chinese yew Thick many candies H-600 groups (600mg/kg) have the effect of anti-trioxypurine and depressor muscle acid anhydride
Fruit, and have protective effect to kidney.
9 southerm yew Thick many candies clinical test of embodiment
A) present invention reduces facing for hyperuricemia by random, control, blind clinical test, evaluation Chinese yew Thick many candies
Bed curative effect and safety.
B) 120 Patients with Hyperuricemia of present invention selection, are randomly divided into control group and treatment group, every group each 60, into
The treatment and observation of 4 weeks behavior phases.Two groups are given Primary Care (diet, movement etc.), and treatment group's medicinal preparation for oral administration is thick containing Chinese yew
Polysaccharide 5gd-1(being calculated with human body weight 70kg), control group takes placebo, by comparing two groups of pretherapy and post-treatment tcm clinical practice diseases
Shape, serum uric acid, blood fat (TC, TG, LDL-C), fasting blood-glucose index, evaluate the clinical efficacy of this product；Before observing two groups of treatments
Blood routine, liver function (ALT, AST), renal function (BUN, Cr), electrocardiogram and adverse reaction afterwards, evaluate its safety.
C) treatment group of the present invention is after 4 weeks, TCM syndrome total effective rate 84%, (the P ＜ of control group total effective rate 25%
0.001)；Treatment group's serum uric acid reduces, compared with before treatment, P<0.01, compared with after control group treatment, P ＜
0.05；Treatment group's patient's LDL-C levels reduce, compared with before treatment, P ＜ 0.05；Two groups of pretherapy and post-treatment blood routines, liver functions
(ALT, AST), renal function (BUN, Cr), electrocardiogram compare, no significant difference (P>0.05)；Equal nothing during two groups of observations
Adverse reaction and serious adverse events occur.
D) conclusion of the present invention：It is curative for effect with Chinese yew Thick many candies treatment hyperuricemia, and high lithemia can be obviously improved
The tcm clinical practice symptom of mass formed by blood stasis can safely and effectively reduce uric acid.
E) data cases of the present invention are as follows：
5 therapeutic effect of table
* * P ＜ 0.001, compared with the control group
6 two groups of reduction serum uric acid curative effects of table compare
* P ＜ 0.01, compared with the control group
The comparison of 7 two groups of pretherapy and post-treatment serum uric acids of table
* P ＜ 0.01, compared with before treatment；#P ＜ 0.05, compared with the control group
Comparison (the cB/mmolL of pretherapy and post-treatment TC, TG, LDL-C, GLU level of 8 two groups of table-1)
* P ＜ 0.05, compared with before treatment
9 two groups of pretherapy and post-treatment blood routines of table compare
The pretherapy and post-treatment Liver and kidney function Indexes Comparison of 10 two groups of table
Show that southerm yew Thick many candies of the present invention have good safety by embodiment 6,7；It is obtained by embodiment 8
Going out the Thick many candies has good anti-trioxypurine effect；It is obtained by embodiment 9, which has good application in clinic.
Claims (9)
1. a kind of preparation method with the yew amylose for dropping high lithemia effect, which is characterized in that include the following steps：
A it) pre-processes
The dry waste material of Chinese yew after extracted complete taxol and 10-DABIII is taken, is crushed, ethyl alcohol is added to carry out Soxhlet extraction,
It is spare that the dregs of a decoction are collected by filtration；
B) water extract-alcohol precipitation
Step A) described in the dregs of a decoction appropriate purified water is added, stirring to the dregs of a decoction is waterlogged, and Soxhlet extraction filters, and is concentrated under reduced pressure；
Ethyl alcohol, stand at low temperature is added in concentrate, and filtration obtains extract；
C it) washs
Step B) in it is extract obtained be slowly added dropwise absolute ethyl alcohol, acetone, ethyl acetate successively, it is dry after crude product；
D) deproteinized
Step C) in gained crude product be dissolved in appropriate hot water, with Sevage methods be added n-butanol/petroleum ether mixed liquor, stirring,
A layer aqueous is removed in centrifugation, repeats aforesaid operations 2-4 times, obtains the polysaccharide solution after removing protein；
E it) dialyses
Step D) in gained polysaccharide solution, using flowing water dialysis remove solution small molecular substance；
F it) is freeze-dried
Step E) in solution decompression concentration after dialysis, be freeze-dried after precooling, obtain Chinese yew Thick many candies；
Step A) in plus ethyl alcohol carry out Soxhlet extraction solid-liquid ratio be 1:8-12.
2. a kind of preparation method with the yew amylose for dropping high lithemia effect as described in claim 1, which is characterized in that
Step A) in smash the dry waste material of Chinese yew mesh number be 50-100 mesh, concentration of alcohol 75-90%, add ethyl alcohol carry out Soxhlet extraction
Solid-liquid ratio is 1:10, extraction time 1-5h；Step B) water extraction twice is carried out, it is 1 that purified water solid-liquid ratio is added for the first time:8-
12, it is 1 that second, which is added purified water solid-liquid ratio,:5-10, step B) in be concentrated under reduced pressure into a concentration of 0.8-1.5g/mL of crude drug；It is dense
3-6 times of volume ethanol is added in contracting liquid, and the temperature of concentration of alcohol 85-97%, stand at low temperature are 2-8 DEG C, time of repose 15-36h.
3. a kind of preparation method with the yew amylose for dropping high lithemia effect as described in claim 1, which is characterized in that
Step C) in be added absolute ethyl alcohol, acetone, ethyl acetate volume be that southerm yew feeds intake 0.05-0.2 times of quality；Step
D n-butanol/petroleum ether mixed liquor of 0.5-2 times of volume) is added, the volume ratio of n-butanol/petroleum ether is 1:3-5.
4. a kind of preparation method as described in claim 1 with the yew amylose for dropping high lithemia effect, which is characterized in that step
Rapid E) dialysis time be 2-5 days, the dialysis bag retention molecular weight 8000-14000 that middle dialysis uses；Solution after dialysis subtracts
Pressure is concentrated into crude drug concentration 2.90-3.10g/mL.
5. a kind of preparation method with the yew amylose of drop high lithemia effect according to any one of claims 1-4 is made
Application of the standby obtained yew amylose in drop high lithemia.
6. a kind of preparation method with the yew amylose of drop high lithemia effect as claimed in claim 5 is preparation-obtained
Application of the yew amylose in drop high lithemia, which is characterized in that yew amylose is prepared as main active for high urine
The drug of acidaemia, drug are the various preparations pharmaceutically received.
7. a kind of preparation method with the yew amylose of drop high lithemia effect according to any one of claims 1-4 is made
The medicament for hyperuricemia that standby obtained yew amylose is prepared, using yew amylose as chief active at
Point, the dosage form of medicament includes：Enema, capsule, oral solution, mouth containing agent, granule, electuary, pill, pill, dissipates tablet
Agent, paste, sublimed preparation, suspension, pulvis, solution, suppository, liniment, creme, spray, powder spray, aerosol, drops, ingot
Agent, patch.
8. a kind of preparation method with the yew amylose of drop high lithemia effect as claimed in claim 7 is preparation-obtained
The medicament for hyperuricemia that yew amylose is prepared, which is characterized in that the auxiliary material for preparing medicament include adhesive,
One kind in filler, diluent, tablet agent, lubricant, disintegrant, colorant, flavoring agent, wetting agent and surfactant or
It is a variety of.
9. a kind of preparation method with the yew amylose of drop high lithemia effect as claimed in claim 7 is preparation-obtained
The medicament for hyperuricemia that yew amylose is prepared, which is characterized in that in medicament yew amylose activity at
Divide content in 1%-99.5%.
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| CN110511290A (en) * | 2018-05-07 | 2019-11-29 | 武汉紫杉红技术开发有限公司 | A kind of preparation method and applications of the yew amylose with drop high lithemia effect |
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| CN112023019A (en) * | 2020-09-15 | 2020-12-04 | 上海健康医学院 | Medicine containing active polysaccharide with effect of reducing uric acid and preparation method thereof |
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| CN101735330A (en) * | 2009-10-12 | 2010-06-16 | 于荣敏 | Preparation method of yew polysaccharide with function of resisting tumors and application thereof |
| CN102397314B (en) * | 2011-11-07 | 2013-08-21 | 赵冬迎 | Health-care product for preventing and treating renal disease of newborn Tibetan mastiff in low-altitude region |
| CN104829743B (en) * | 2015-05-25 | 2017-07-11 | 萧丽雅 | A kind of preparation method and its usage of Polysaccharides from Leaves of Moringa oleifera |
| CN108586628A (en) * | 2018-05-07 | 2018-09-28 | 武汉紫杉红技术开发有限公司 | A kind of preparation method and applications with the yew amylose for dropping high lithemia effect |
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Application publication date: 20180928 |