CN104592411B - Phosphoric acid esterification snakegourd peel polysaccharide and medical application thereof - Google Patents
Phosphoric acid esterification snakegourd peel polysaccharide and medical application thereof Download PDFInfo
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Abstract
The invention discloses phosphoric acid esterification snakegourd peel polysaccharide and a medical application thereof. The phosphoric acid esterification snakegourd peel polysaccharide is characterized by being prepared by performing phosphoric acid esterification modification on snakegourd peel polysaccharide by using sodium tripolyphosphate and sodium trimetaphosphate. The phosphoric acid esterification snakegourd peel polysaccharide disclosed by the invention has a significant anti-ageing function, does not have toxic or side effects, cannot cause damages to internal organs, and can be used for preparing anti-ageing medicaments.
Description
Technical field
The present invention relates to a kind of modified Pericarpium Trichosanthis polysaccharide, be specifically related to a kind of Phosphation Pericarpium Trichosanthis polysaccharide and medical usage thereof.
Background technology
Aging is the process that body is respectively organized, organ dysfunction increases with the age and occurs degeneration to change.According to " ethnic drug Neo-Confucianism is miscellaneous
Will " (Journal of Ethnopharmacology, 2010,132:512-517) introduction, the increase of living radical can damage body
Tissue or cell, the damage accumulation caused when free radical has exceeded the repair ability of body, can accelerate the process of body aging.
Along with the quickening of world's most countries aged tendency of population trend, increasing scholar is absorbed in research body antioxidation, anti-ageing
The research of old acting formulations.Therefore, from natural product, find the antiaging agent having no side effect to have great importance.
Vegetable polysaccharides is by the biomacromolecule material of multiple monosaccharide molecule condensation, is prevalent in plant, including forming sediment
Powder, cellulose, polysaccharide, pectin etc..Vegetable polysaccharides is soluble in water, and its biological activity is extensive, has regulation immunologic function,
The biological activitys such as blood sugar lowering, suppresses tumor, anti-bacteria and anti-virus.
Chinese juniper beach wormwood (Trichosanthes kirilowii Maxim) is the perennial liana of one that Cucurbitaceae Chinese juniper beach wormwood belongs to, and is distributed widely in
China Yangtze river basin and northern area thereof.The crust of the mature fruit of Chinese juniper beach wormwood is referred to as Pericarpium Trichosanthis.Pericarpium Trichosanthis is rich in there being abundant amino
Acid, mineral element and active polysaccharide etc., have higher medical value.As China " Chinese Journal of New Drugs " (2010,20:
1872-1874), " Shanxi Medicine magazine " (2009,38:67-68) and " China Dispensary " (2009,20:648-651) describes
Pericarpium Trichosanthis injection has the effect for the treatment of stable angina pectoris, and Pericarpium Trichosanthis decoct is to dysentery bacterium, streptococcus pneumoniae, hemolytic chain
Coccus and diphtheria corynebacterium etc. all have inhibitory action, can improve immune function of mice；China " Chinese pharmacists " (2004,7:562-563)
Describe employing water-bath extraction separates from Pericarpium Trichosanthis obtain one water soluble polysaccharide, this polysaccharide is by rhamnose, arabinose
Forming with glucose, its mol ratio is: 1.0:2.4:4.7；China " HeFei University of Technology's report (natural science edition) "
(2014,03:359-363) describe microwave radiation exaraction Pericarpium Trichosanthis polysaccharide and there is stronger antioxidant activity.China Patent No.
201310133957.9 disclose a kind of Pericarpium Trichosanthis polysaccharide composite capsule and preparation method thereof, use with Fructus Trichosanthis polysaccharide, the Rhizoma Anemarrhenae Radix Rehmanniae
Composite powder is primary raw material and is aided with other materials, prepares a kind of fructus trichosanthis polysaccharide composite capsule, this polysaccharide composite capsule at normal temperatures
Easily it is absorbed by the body, Fructus Trichosanthis, the Radix Rehmanniae, the nutritive value of the Rhizoma Anemarrhenae and medical value can be played fully.
The research being modified Pericarpium Trichosanthis polysaccharide the most both at home and abroad is less.There are no open source literature both at home and abroad at present to mention Fructus Trichosanthis
Skin polysaccharide takes Phosphation to modify to be modified as Phosphation polysaccharide, and as preparing the application aspect of antiaging agent
Report.
Summary of the invention
It is an object of the invention to provide a kind of Pericarpium Trichosanthis polysaccharide taking Phosphation modification and preparing antiaging agent
In application.
The present invention solves technical problem, adopts the following technical scheme that
The Phosphation Pericarpium Trichosanthis polysaccharide of the present invention, its feature is: described Phosphation Pericarpium Trichosanthis polysaccharide is by with trimerization phosphorus
Acid sodium and sodium trimetaphosphate carry out Phosphation modification acquisition to Pericarpium Trichosanthis polysaccharide.
The Phosphation Pericarpium Trichosanthis polysaccharide of the present invention is prepared as follows:
Sodium tripolyphosphate and sodium trimetaphosphate 4.5-5.5:1 in mass ratio are mixed and constitutes phosphorylation agent；By 12-13g phosphoric acid
Change reagent to be dissolved in distilled water, add 1.0-1.2g sodium sulfate and add distilled water and be settled to 100mL, adding 1.00g Pericarpium Trichosanthis
Polysaccharide, then with concentration for 0.1mol L-1NaOH solution regulation pH to 7.5-8, it is thus achieved that reaction system；By reaction system
Reacting 6.5-8.0h in 65-70 DEG C of water bath with thermostatic control, be subsequently adding the dehydrated alcohol of 4-4.5 times of volume, precipitate with ethanol 24h being centrifuged obtains
Obtain precipitate with ethanol polysaccharide；Precipitate with ethanol polysaccharide is redissolved after vacuum lyophilization in 60 DEG C of water, be 13000Da then at molecular cut off
Bag filter is first dialysed 24-48h with tap water reverse flow, then dialyses 24-48h with distilled water, in collecting bag filter, retain liquid
And rotary evaporation concentrates, after lyophilization, it is thus achieved that Phosphation Pericarpium Trichosanthis polysaccharide.
The present invention further discloses the medical usage of above-mentioned Phosphation Pericarpium Trichosanthis polysaccharide, its feature is: Phosphation Fructus Trichosanthis
The application in preparing antiaging agent of the skin polysaccharide.The present invention is by the effect pharmacological evaluation to exhausted mining areas defying age, really
Found the anti-aging effects of Phosphation Pericarpium Trichosanthis polysaccharide.
The Phosphation Pericarpium Trichosanthis polysaccharide of the present invention will be able to be had by the customary preparation methods of pharmaceutics when preparing antiaging agent
This Phosphation Pericarpium Trichosanthis polysaccharide of effect amount and pharmaceutic adjuvant hybrid process, be prepared as oral tablet or capsule.
Compared with the prior art, beneficial effects of the present invention is embodied in:
The Phosphation Pericarpium Trichosanthis polysaccharide of the present invention has significant anti-senescence function, and has no side effect, will not be to intracorporeal organ
Cause damage.The inventive method simple economy, can be applied to prepare tablet or capsule formulation by this Phosphation Pericarpium Trichosanthis polysaccharide
Antiaging agent.
Accompanying drawing explanation
Fig. 1 is the infrared spectrogram of the Pericarpium Trichosanthis polysaccharide (TPP-1) before Phosphation modification；
Fig. 2 is that to use sodium tripolyphosphate and sodium trimetaphosphate that Pericarpium Trichosanthis polysaccharide carries out the Phosphation Pericarpium Trichosanthis after modification many
The infrared spectrogram of sugar (PTPP-1)；
Fig. 3 is that Pericarpium Trichosanthis polysaccharide TPP-1 (A in Fig. 3) before modified and Phosphation Pericarpium Trichosanthis polysaccharide PTPP-1 is (in Fig. 3
B)13C NMR spectra figure；
Fig. 4 is Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1)31P NMR spectra figure.
Detailed description of the invention
Below by embodiment and combine accompanying drawing the present invention is made further concrete detailed description.
Embodiment 1
One, the preparation of Pericarpium Trichosanthis polysaccharide (TPP-1):
The present embodiment prepares TPP-1 the most as follows:
Pericarpium Trichosanthis cleans up, dries and pulverizes acquisition Fructus Trichosanthis corium farinosum；Petroleum ether and anhydrous second it is sequentially added in Fructus Trichosanthis corium farinosum
Alcohol, backflow 8h carries out preliminary defat and decolouring respectively, the most naturally dries acquisition Pericarpium Trichosanthis dry powder；Matter by Pericarpium Trichosanthis dry powder
Amount is 1:35mg/mL with the volume ratio of distilled water, addition distilled water in Pericarpium Trichosanthis dry powder, employing ultrasonic temperature 60 DEG C,
Ultrasonic time 30min, the ultrasonic wave added water-bath restricted-access media 3 times of ultrasonic power 300W, be filtered to remove filtering residue, merges filter
Liquid obtains Pericarpium Trichosanthis extracting solution；
Pericarpium Trichosanthis extracting solution is concentrated by application Rotary Evaporators (Asia, Shanghai is flourish), after being centrifuged, adds 95% ethanol of 4 times of volumes,
4 DEG C of precipitate with ethanol 12-24h, then 4000r min-1Centrifugal 10min, it is thus achieved that precipitate with ethanol polysaccharide；Use distilled water to dissolve precipitate with ethanol polysaccharide to obtain
To Pericarpium Trichosanthis crude polysaccharides solution, lyophilization obtains Pericarpium Trichosanthis polysaccharide crude.
Again this Pericarpium Trichosanthis polysaccharide crude distilled water is redissolved, use Sai Wojiefa (Sevag) deproteination (acutely to shake 20min
It is placed on static 20min in separatory funnel, separate aqueous layer and the Denatured protein of solvent layer intersection, repeats to aqueous phase and solvent phase
Interface without gluey denatured protein), add 30% hydrogen peroxide constant temperature decolouring.By the Pericarpium Trichosanthis polysaccharide after deproteinization, decolouring
Crude product aqueous solution is placed in the bag filter that molecular cut off is 13000Da, and tap water reverse flow dialysis 48h, distilled water is dialysed
24h.Freeze-dried Pericarpium Trichosanthis polysaccharide (TPP-1).Gained Pericarpium Trichosanthis polysaccharide is 1.0:3.3:4.3:6.0's by mol ratio
L-arabinose, D-MANNOSE, D-Glucose and D-galactose composition.
Two, the preparation of Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1)
The Pericarpium Trichosanthis polysaccharide that the present embodiment method described above is obtained is raw material, is prepared as follows Phosphation Pericarpium Trichosanthis polysaccharide:
9.82g sodium tripolyphosphate and 2.18g sodium trimetaphosphate are dissolved in distilled water, add 1.2g sodium sulfate and add distilled water calmly
Hold to 100mL, add 1.00g TPP-1, then with concentration for 0.1mol L-1NaOH solution regulation pH to 8,
Obtain reaction system；Reaction system is reacted in 70 DEG C of waters bath with thermostatic control 6.5h, is subsequently adding 95% ethanol of 4 times of volumes,
Precipitate with ethanol 24h centrifugal acquisition precipitate with ethanol polysaccharide；Precipitate with ethanol polysaccharide is redissolved after vacuum lyophilization in 60 DEG C of water, then at retaining
Molecular weight is first to use tap water reverse flow dialysis 36h in 13000Da bag filter, then (uses distillation water logging with distilled water dialysis
Bubble also changes distilled water every 4h) 24h, retain liquid in collecting bag filter and rotary evaporation concentrates, after lyophilization, obtain phosphorus
Esterification Pericarpium Trichosanthis polysaccharide PTPP-1.
Phosphate content in test product phosphoric acid esterification Pericarpium Trichosanthis polysaccharide as follows:
Phosphate radical Specification Curve of Increasing and phosphate radical measure what reference China " Food Science " (2011,32 (24): 73-77) was reported
Method.
Phosphate radical Specification Curve of Increasing: draw respectively 0mL, 0.5mL, 1mL, 1.5mL, 2mL, 2.5mL, 3mL,
3.5mL, 4mL, 4.5mL and 5mL concentration is the KH of 0.1mg/mL2PO4Standard solution, in test tube, supplements distillation
Water, to 5mL, adds 3mL tris buffer solution, takes the V that 1mL mass concentration is 20% respectivelyCAqueous solution, 1mL are dense
Degree is 3mol L-1H2SO4Join after solution and ammonium molybdate aqueous solution mix homogeneously that 1mL mass concentration is 3%
KH2PO4In the mixed solution of standard solution and tris buffer solution, after reacting 30min at 45 DEG C, in 580nm (752 points
Light photometer) place's mensuration absorbance, with absorbance as vertical coordinate, phosphate concentration is abscissa, draws phosphate radical standard curve.
Phosphate radical measures: takes 0.1g sample and adds heating in the test tube filling 1mL concentrated sulphuric acid and 1mL concentrated nitric acid, after cooling
Add 1mL H2O2Solution (30%), is slowly heated, and repeat above step to reaction sample liquid be faint yellow or water white transparency.
Adding 1mL concentration is that after the heating of 6mol/L hydrochloric acid makes acid thoroughly reaction, constant volume is in 50mL volumetric flask.According still further to above-mentioned phosphorus
Acid group standard curve method measures absorbance, and calculates phosphate content according to standard curve.
In above-mentioned test understands the present embodiment gained Phosphation Pericarpium Trichosanthis polysaccharide, phosphate radical mass fraction is 14.36%.
Three, the structure verification of Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1)
Infrared spectrum measurement: take above-mentioned Pericarpium Trichosanthis polysaccharide 2mg before modified respectively, with employing sodium tripolyphosphate and sodium trimetaphosphate
Phosphation Pericarpium Trichosanthis polysaccharide 2mg prepared by modification, with the quality (mg) of polysaccharide and quality (mg) ratio of KBr for 1 30,
TPP-1 or PTPP-1 is mixed homogeneously with KBr, tabletting after grinding, use Fourier transform infrared spectrometer (Nexus 670)
Carrying out IR spectrum analysis, sweep limits is 4000-400cm-1。
Fig. 1 is the infrared spectrogram of Pericarpium Trichosanthis polysaccharide (TPP-1) before modified；Fig. 2 is for using sodium tripolyphosphate and three Metaphosphoric acids
Sodium carries out the infrared spectrogram of Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1) after modification to Pericarpium Trichosanthis polysaccharide.Relative analysis this
The result of two infrared spectrograms shows, before modified after Pericarpium Trichosanthis polysaccharide be respectively provided with the characteristic absorption peak of Pericarpium Trichosanthis polysaccharide, from figure
At PTPP-1 at 1140cm in 2-1The stretching vibration peak at place shows, PTPP-1 contains pyranoid form glycosidic bond.Additionally, 2920cm-1
-the CH at place2Stretching vibration absworption peak, 1640cm-1The stretching vibration absworption peak of the C=O at place and 1410cm-1The C-O's at place
, the most there is not significant change in stretching vibration absworption peak before and after modifying, illustrates that Phosphation modifies the knot not destroying TPP-1
Structure；In Fig. 2, the infrared spectrum of PTPP-1 shows, at 1232cm-1Place is the characteristic absorption peak caused by P=O stretching vibration,
Another is at 976cm-1Place is the characteristic absorption peak caused by P-O-C stretching vibration, and this result shows sodium tripolyphosphate and three inclined
Sodium phosphate success polysaccharide-modified to Pericarpium Trichosanthis.
Nuclear magnetic resonance spectrometry: take 50mg TPP-1 and PTPP-1 respectively and be dissolved in 1.0mL D2In O, use Bruker
AV-500 nuclear magnetic resonance spectrometer is carried out13C NMR analyzes.Take 50mg PTPP-1 and be dissolved in 1.0mL D2In O, use Bruker
AV-500 nuclear magnetic resonance spectrometer is carried out31P NMR analyzes.
Fig. 3 is Pericarpium Trichosanthis polysaccharide TPP-1 (A) before modified and Phosphation Pericarpium Trichosanthis polysaccharide PTPP-1 (B)13C NMR spectra
Figure.Fig. 3 (A) C compared with (B)1、C2、C3、C5Chemical shift all do not occur significantly to change, and Phosphation Pericarpium Trichosanthis is many
Sugar is at C4、C6Chemical shift at signal peak be obviously reduced, this result is due to-H2PO3Group and C4And C6Hydroxyl
Cause in conjunction with caused.
Fig. 4 is Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1)31P NMR spectra figure.As shown in Figure 4,0.946～1.727ppm
Between, phosphated derivant shows strong multiple signal formant, and result shows, phosphorylation modification success, and phosphoric acid
Group is positioned in several different magnetic field environment, thus instead of the diverse location in polysaccharide molecule chain.
Above-mentioned infrared absorption spectroscopy and nuclear magnetic resonance result show: the phosphorylation modification success of Pericarpium Trichosanthis polysaccharide, and the phosphorus modified
Acid groups is connected to C4And C6On.
Four, Pericarpium Trichosanthis polysaccharide (TPP-1) and the anti-aging effects pharmacological evaluation of Phosphation Pericarpium Trichosanthis polysaccharide (PTPP-1):
(1) experimental raw: prepare the normal saline solution of TPP-1 and PTPP-1:
The normal saline that TPP-1 mass percentage concentration is 0.9% is become the polysaccharide solution of 10mg/mL and 40mg/mL,
As low concentration TPP-1 normal saline solution and high concentration TPP-1 normal saline solution；
By molten for polysaccharide that the normal saline that PTPP-1 mass percentage concentration is 0.9% becomes 10mg/mL and 40mg/mL
Liquid, as low concentration PTPP-1 normal saline solution and high concentration PTPP-1 normal saline solution；
(2) experiment animal used: kunming mice 70, male, body weight 20 ± 2g, SPF level, it is randomly divided into 7 groups.
(3) experiment used kit: superoxide dismutase (SOD) testing cassete, catalase (CAT) test kit,
Glutathion peroxidase (GSH-PX) test kit and malonaldehyde (MDA) test kit, build up biological engineering for Nanjing and grind
Study carefully produced product.
(4) experimental technique:
First, after 70 healthy kunming mice adaptabilities being fed one week, weigh.Take 60 healthy kunming mices, lumbar injection
Concentration is D-galactose (Sigma Co., USA) aqueous solution (dosage 100mg/kg) of 2%, once a day, injects continuously
After 6 weeks, mice lethargy is depressed, body slight of stature, and presents the oldest and the most feeble feature, shows that D-gal exhausted mining areas is built
Mould success；Simultaneously to 10 normal kunming mice lumbar injection 0.9% normal saline (dosage 100mg/kg) of residue, 1 time/d,
Injection 6 weeks continuously.
Then, 60 D-gal exhausted mining areas are divided into following six groups:
D-gal model group: every day gavage 10mL kg-1The normal saline of dosage；
Positive controls: every day gavage 100mg kg-1The V of dosageC；
TPP-1 low dose group: every day gavage 100mg kg-1TPP-1 (the i.e. gavage every day low concentration TPP-1 physiology salt of dosage
The dosage of aqueous solution is 10mL kg-1)；
TPP-1 high dose group: every day gavage 400mg kg-1TPP-1 (the i.e. gavage every day high concentration TPP-1 physiology salt of dosage
The dosage of aqueous solution is 10mL kg-1)；
PTPP-1 low dose group: every day gavage 100mg kg-1(i.e. gavage every day low concentration PTPP-1 is raw for the PTPP-1 of dosage
The dosage of reason saline solution is 10mL kg-1)；
PTPP-1 high dose group: every day gavage 400mg kg-1(i.e. gavage every day high concentration PTPP-1 is raw for the PTPP-1 of dosage
The dosage of reason saline solution is 10mL kg-1)；
Additionally, using 10 normal kunming mices as Normal group, every day gavage 10mL kg-1The normal saline of dosage)；
By above seven groups of mice timing every day according to dosage gavages, 1 time/d, continuous 30 days, each group was often daily freely drunk water and takes food
Standard feed.During gavage, observe the situations such as the mice mental status, body every day；After last gavage, water 12h is can't help in fasting,
After weighing, pluck eyeball and take blood and be placed in the centrifuge tube that anticoagulant processes, be centrifuged 3min with the rotating speed of 5000rpm and obtain serum and put
Standby in-20 DEG C；De-cervical vertebra puts to death mice, takes out mouse spleen, thymus, liver, brain, frozen standby.
(4) index of correlation measures:
Mouse spleen, the mensuration of thymus index；
In mouse liver, brain, serum, SOD enzyme activity, the work of GSH-PX enzyme, CAT enzyme are lived and the mensuration of MDA enzyme work: little
Rathole ball takes blood, and 5000r/min is centrifuged 3min, take serum be put in-20 DEG C standby.Liver, brain, serum recover before use to
Room temperature, operates according to test kit description step, measure SOD enzyme activity, GSH-PX enzyme activity, CAT enzyme live and
MDA content.
(5) the pharmacological results:
Table 1 gives above-mentioned Pericarpium Trichosanthis polysaccharide and Phosphation Pericarpium Trichosanthis polysaccharide antagonism mouse aging body weight thereof and the shadow of organ index
Ring.
Table 1 TPP-1 and PTPP-1 antagonism mouse aging body weight and the impact of organ index
Note: compared with D-gal model group,aP < 0.05,bp<0.01。
Each group index parameter in table 1 is compared analysis, it can be seen that TPP-1 and PTPP-1 is to each process group mice phase
Close the impact of index.D-gal model group mice lethargy, fur is matt, and perpendicular hair phenomenon is obvious, and activity significantly reduces,
PTPP-1, TPP-1 impact such as table 1 on exhausted mining areas body weight and organ index:
From table 1 it follows that compared with D-gal model group mice, TPP-1, PTPP-1 body weight to exhausted mining areas
All there is increase in various degree, and PTPP-1 high dose group body weight increases more notable, illustrates what D-gal was caused by PTPP-1
Mouse Weight increasess slowly the effect of increasing significantly；Compared with D-gal model group, TPP-1, PTPP-1 are little to aging model
The thymus index of Mus, index and spleen index all have the thymus of rising in various degree, PTPP-1 high dose group mice and low dose group mice
Index, index and spleen index the most relatively TPP-1 group is high, shows the aging of the notable slow down aging model mice immune organ of PTPP-1 energy,
Strengthen its immunologic function.
Table 2 gives SOD, GSH-PX, CAT activity in TPP-1 and PTPP-1 antagonism mouse aging liver, brain, serum
And the impact of MDA content.
SOD, GSH-PX, CAT activity and the impact of MDA content in table 2 TPP-1 and PTPP-1 antagonism mouse aging liver, brain, serum
Note: compared with model group,aP < 0.05,bp<0.01。
Each group index parameter in table 2 is compared analysis, it can be seen that TPP-1 and PTPP-1 refers to each group of mice is relevant
Target affects.As shown in table 2, compared with D-gal model group mice, TPP-1, PTPP-1 group and positive controls mice
Liver, brain and serum in SOD, GSH-PX, CAT activity is notable raises, MDA content significantly drops, and low D-gal exists
Metabolic process can make body produce the free radical of excess, form lipid peroxide, destroy body Antioxidative Defense System.Its
Middle PTPP-1 high dose group liver homogenate, brain homogenate and activity of SOD in serum add 49.55%, 33.16% and 31.23% respectively,
GSH-PX activity adds 21.68%, 21.91% and 12.97% respectively, and CAT activity adds 56.56%, 46.45% respectively
With 28.22%, MDA content reduces 26.50%, 34.14% and 33.63% respectively.Description of test, Pericarpium Trichosanthis polysaccharide has
Significantly activity of fighting against senium, and the Phosphation polysaccharide after sodium tripolyphosphate and sodium trimetaphosphate modification of same dose
Defying age ability is higher, shows that Phosphation modifies the defying age ability that improve this polysaccharide.
Above-mentioned pharmacological experiment analysis determines the most representational index of mice age, as can be seen from the above results, is being administered
When dosage is 100-400mg/kg, Pericarpium Trichosanthis polysaccharide has significant anti-aging effects, is remarkably improved the body of exhausted mining areas
Weight and organ index, be effectively increased SOD, GSH-PX, CAT enzyme and live, reduce MDA content, have obvious defying age
Activity.And the defying age of the Phosphation Pericarpium Trichosanthis polysaccharide after sodium tripolyphosphate and sodium trimetaphosphate modification of same concentrations
Activity is remarkably reinforced.
Five, the processing of Phosphation Pericarpium Trichosanthis polysaccharide formulation
1, oral tablet processing
After the Phosphation Pericarpium Trichosanthis polysaccharide of 200-350g is mixed with pharmaceutic adjuvant diluent 200-300g, fluidizer 20-50g,
Mixing homogeneously with 200-400mL ethanol and prepare soft material, soft material sieves and obtains wet granular, and dried granulate is eventually adding lubricant
Tabletting after 5-10g mixing, makes 1000 tablets.Described pharmaceutic adjuvant diluent selects dextrin or dried starch；Described fluidizer
Use Pulvis Talci, micropowder silica gel or microcrystalline Cellulose；Described lubricant uses magnesium stearate or zinc stearate.
The preference of the tablet processing that the present embodiment is recommended is: took the above-mentioned Phosphation Pericarpium Trichosanthis polysaccharide 300 of 80-100 mesh sieve
G, diluent dextrin 200g, fluidizer Pulvis Talci 40g, add 400mL ethanol mix homogeneously and prepare soft material after mix homogeneously, soft
Material sieves and obtains wet granular, dried granulate, is eventually adding tabletting after magnesium stearate lubricant 8g mixing, makes 1000,
Tablet weight 0.94g, polyoses content 0.3g/ sheet.
2, capsule processing
After pharmaceutic adjuvant diluent 200-300g, fluidizer 50-70g, wetting agent 2-6g mixing, then with 300-600g's
Above-mentioned Phosphation Pericarpium Trichosanthis polysaccharide mix homogeneously, fills 1# Capsules, prepares capsule 1000.Described pharmaceutic adjuvant is dilute
Release agent and select dextrin or dried starch；Described fluidizer uses Pulvis Talci or micropowder silica gel or microcrystalline Cellulose；Described wetting agent uses
Sodium lauryl sulphate or dioctyl sulphosuccinate.
The capsule processing preference that the present embodiment is recommended is: take diluent dried starch 200g, fluidizer Pulvis Talci 40g, moistening
After agent sodium lauryl sulphate 5g mix homogeneously, then Phosphation Pericarpium Trichosanthis polysaccharide above-mentioned with 400g mix homogeneously after fill 1# hollow
Capsule 1000, polyoses content 0.4g/ grain.
Embodiment 2
The present embodiment prepares TPP-1 the most as follows:
Being cleaned up by Pericarpium Trichosanthis, dry, pulverize, be separately added into petroleum ether and ethanol, backflow 10h carries out preliminary defat respectively
And decolouring, dry for 60 DEG C and obtain Pericarpium Trichosanthis dry powder；By being 1 30mg/ with the volume ratio by the quality of Pericarpium Trichosanthis dry powder Yu distilled water
ML, adds distilled water in Pericarpium Trichosanthis dry powder, uses ultrasonic temperature 55 DEG C, ultrasonic time 40min, ultrasonic power 350W
Ultrasonic wave added water-bath restricted-access media 3 times；It is filtered to remove filtering residue and obtains Pericarpium Trichosanthis extracting solution；
Pericarpium Trichosanthis extracting solution rotary evaporation is concentrated, after being centrifuged, adds 95% ethanol of 3 times of volumes, 4 DEG C of precipitate with ethanol 12h；So
Rear 4000r min-1Centrifugal 15min, incline supernatant, it is thus achieved that precipitate with ethanol polysaccharide；Use distilled water to dissolve precipitate with ethanol polysaccharide and obtain Fructus Trichosanthis
Skin crude polysaccharides solution, lyophilization obtains Pericarpium Trichosanthis polysaccharide crude.
Again this Pericarpium Trichosanthis polysaccharide crude distilled water is redissolved, use Sai Wojiefa (Sevag) deproteinization, add 30% dioxygen
45 DEG C of constant temperature decolourings of water.Pericarpium Trichosanthis polysaccharide crude aqueous solution after deproteinization, decolouring is placed in molecular cut off is 13000Da
Bag filter in, tap water reverse flow dialysis 48h, then dialyse 24h with distilled water.Freeze-dried Pericarpium Trichosanthis polysaccharide
(TPP-1)。
The Pericarpium Trichosanthis polysaccharide that the present embodiment method described above is obtained is raw material, is prepared as follows Phosphation Pericarpium Trichosanthis polysaccharide:
11.00g sodium tripolyphosphate and 2.00g sodium trimetaphosphate are mixed and constitutes phosphorylation agent；By 12-13g phosphorylation agent
It is dissolved in distilled water, adds 1.0g sodium sulfate and add distilled water and be settled to 100mL, adding 1.00g Pericarpium Trichosanthis polysaccharide, so
After with concentration for 0.1mol L-1NaOH solution regulation pH to 7.8, it is thus achieved that reaction system；By reaction system 65 DEG C of perseverances
Tepidarium is reacted 8h, adds 95% ethanol of 4.5 times of volumes, precipitate with ethanol 20h centrifugal acquisition precipitate with ethanol polysaccharide.Precipitate with ethanol polysaccharide
Redissolve after vacuum lyophilization in 60 DEG C of water, then at molecular cut off be in 13000Da bag filter first reverse with tap water
Flowing dialysis 24h dialyses 24h with distilled water again, collects and concentrates solution, after lyophilization, obtain Phosphation Pericarpium Trichosanthis many
Sugar (PTPP-1).By the method in embodiment 1, Phosphation Pericarpium Trichosanthis polysaccharide is carried out phosphate content mensuration, its phosphate radical matter
Amount mark is 13.59%.Similar to the phosphate content in embodiment 1.
Then above-mentioned Pericarpium Trichosanthis polysaccharide and normal saline that Phosphation polysaccharide mass percentage concentration is 0.9% thereof are prepared respectively
Becoming the aqueous solution of 5mg/mL and 10mg/mL, being administered volume is 10mL kg-1。
Pericarpium Trichosanthis polysaccharide and the Phosphation polysaccharide thereof of above-mentioned each dosage are carried out exhausted mining areas anti-aging effects pharmacological evaluation,
The mode step used and other conditions are the most in the same manner as in Example 1, have also been obtained the result similar with embodiment 1.
The description of test of above-described embodiment, Pericarpium Trichosanthis polysaccharide and the phosphate ester after sodium tripolyphosphate and sodium trimetaphosphate modification
Change Pericarpium Trichosanthis polysaccharide and be respectively provided with significant anti-senescence function, and have no side effect, its preparation method simple economy, therefore can be by Fructus Trichosanthis
Phosphation Pericarpium Trichosanthis polysaccharide after skin polysaccharide and employing Phosphation modification, by the customary preparation methods of pharmaceutics, by effective dose
Phosphation Pericarpium Trichosanthis polysaccharide and appropriate pharmaceutic adjuvant hybrid process be prepared as the Kangshuaining mixture of oral tablet or capsule formulation
Thing.
Claims (3)
1. a Phosphation Pericarpium Trichosanthis polysaccharide, it is characterised in that: described Phosphation Pericarpium Trichosanthis polysaccharide is by with tripolyphosphate
Sodium and sodium trimetaphosphate carry out Phosphation modification acquisition to Pericarpium Trichosanthis polysaccharide, are prepared the most as follows:
Sodium tripolyphosphate and sodium trimetaphosphate 4.5-5.5:1 in mass ratio are mixed and constitutes phosphorylation agent；By 12-13g phosphoric acid
Change reagent to be dissolved in distilled water, add 1.0-1.2g sodium sulfate and add distilled water and be settled to 100mL, adding 1.00g Pericarpium Trichosanthis
Polysaccharide, then with concentration for 0.1mol L-1NaOH solution regulation pH to 7.5-8, it is thus achieved that reaction system；By reaction system
Reacting 6.5-8.0h in 65-70 DEG C of water bath with thermostatic control, be subsequently adding the dehydrated alcohol of 4-4.5 times of volume, precipitate with ethanol 24h being centrifuged obtains
Obtain precipitate with ethanol polysaccharide；Precipitate with ethanol polysaccharide is redissolved after vacuum lyophilization in 60 DEG C of water, be 13000Da then at molecular cut off
Bag filter is first dialysed 24-48h with tap water reverse flow, then dialyses 24-48h with distilled water, in collecting bag filter, retain liquid
And rotary evaporation concentrates, after lyophilization, it is thus achieved that Phosphation Pericarpium Trichosanthis polysaccharide.
2. the medical usage of Phosphation Pericarpium Trichosanthis polysaccharide described in a claim 1, it is characterised in that: Phosphation Pericarpium Trichosanthis
Polysaccharide application in preparing antiaging agent.
Medical usage the most according to claim 2, it is characterised in that: prepared antiaging agent is normal by pharmaceutics
Regulation Preparation Method, the oral tablet prepared for active substance with Phosphation Pericarpium Trichosanthis polysaccharide or capsule.
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| CN106432521B (en) * | 2016-09-22 | 2019-05-24 | 上海应用技术大学 | A kind of preparation method of Phosphation bletilla polysaccharide |
| CN108623699B (en) * | 2017-03-17 | 2021-10-08 | 上海医药集团股份有限公司 | Trichosanthes peel polysaccharide GLP-5-1, preparation method and application thereof |
| CN107216404A (en) * | 2017-06-06 | 2017-09-29 | 威海温喜生物科技有限公司 | A kind of alga oligosaccharides phosphate and preparation method thereof |
| CN108530559A (en) * | 2018-04-21 | 2018-09-14 | 潍坊医学院 | A kind of preparation method of marine algae polysaccharide derivant |
| CN110483654B (en) * | 2019-08-27 | 2021-04-27 | 武汉轻工大学 | Preparation method and application of modified huperzia serrata polysaccharide |
| CN114469988B (en) * | 2022-03-01 | 2022-08-16 | 北京天赋神奇科技有限公司 | Biological preparation containing water-soluble fatty acid and trichosanthes bark polysaccharide |
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