CN100362022C - Cuttlebone polysaccharide CPS-1 and its preparation and use - Google Patents
Cuttlebone polysaccharide CPS-1 and its preparation and use Download PDFInfo
- Publication number
- CN100362022C CN100362022C CNB2005101100826A CN200510110082A CN100362022C CN 100362022 C CN100362022 C CN 100362022C CN B2005101100826 A CNB2005101100826 A CN B2005101100826A CN 200510110082 A CN200510110082 A CN 200510110082A CN 100362022 C CN100362022 C CN 100362022C
- Authority
- CN
- China
- Prior art keywords
- cps
- water
- preparation
- elutriant
- cuttlebone polysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The present invention relates to the technical field of medicine, particularly to polysaccharide CPS-1 extracted from cuttlefish bone, a preparation method thereof and an application thereof. The preparation method comprises the following steps: cuttlefish bone powder is boiled with water, and extracting solution is obtained; the extracting solution is deposited with alcohol, the deposited extracting solution is dialyzed, frozen and dried, and a cuttlefish bone polysaccharide crude product is obtained; and a DEAE-Sepharose F. F ion exchange column, a Sephacryl S-300 molecule coagulation column and a Sepharose CL-6B coagulation column are orderly used for separation and purification, and the cuttlefish bone polysaccharide CPS-1 is obtained. Through animal experiments, the polysaccharide CPS-1 has the function of treating ulcerative colitis. Therefore, the polysaccharide CPS-1 can be used for preparing medicaments for treating ulcerative colitis.
Description
Technical field
The present invention relates to medical technical field, is polysaccharide CPS-1-1 that extracts from Endoconcha Sepiae and its production and use.
Background technology
Endoconcha Sepiae has another name called Os Sepiae, is the inner casing of marine animal golden cuttlefish, Sepia andreana or sepiella maindroni de Rochebrune, is the traditional Chinese medicine material.Its property is salty, puckery, warm, astringing to arrest bleeding, relieving leukorrhea by astringents, relieving haperacidity is arranged, hold back effects such as sore, is widely used.Itself is as the waste material of food-processing industry, and minority reclaims as Chinese medicinal materials, and major part is not all well utilized.Endoconcha Sepiae often is used for the treatment of various ulcer with other drug matchings clinical, has better curative effect.But its effective constituent is not determined so far, does not see the report of relevant polysaccharide component yet.
Summary of the invention
The present invention extracts a kind of Cuttlebone polysaccharide with antiulcer action, called after CPS-1 from Endoconcha Sepiae.Through setting up mouse experiment ulcer colitis animal model, find that Cuttlebone polysaccharide CPS-1-1 has the effect of quickening chronic ulcer tissue healing, protection intestinal tissue.Therefore can be used for preparing the medicine for the treatment of ulcerative colitis.
The preparation method of Cuttlebone polysaccharide CPS-1 of the present invention-1 is as follows:
1. prepare water extract solution: the Endoconcha Sepiae solid is ground into powder, adds water boil 8-10 hour, suction filtration obtains filtrate while hot, filtrate is concentrated the centrifugal precipitation of abandoning.Supernatant liquor is that 4: 1 sevage method extraction repeatedly removes albumen with the volume ratio of chloroform and propyl carbinol, does not contain protein to water, obtains water extract solution.
2. preparation Cuttlebone polysaccharide crude product: said extracted thing solution is added ethanol sedimentation spend the night, the centrifugal supernatant of abandoning will precipitate and redissolve in distilled water, and the centrifugal insolubles that goes is with supernatant alcohol precipitation once more, the centrifugal supernatant that goes.The dissolving of precipitation adding distil water, flowing water dialysis at least 36 hours.With the dialyzate lyophilize, get the Cuttlebone polysaccharide crude product.
3. separation and purification, preparation Cuttlebone polysaccharide CPS-1-1: will be dissolved in distilled water in the hot water bath of gained Cuttlebone polysaccharide crude product, centrifugal collection supernatant is dissolved in distilled water in the hot water bath once more with insolubles, the centrifugal precipitation of abandoning.Twice supernatant liquor merged, be splined on DEAE Sepharose F.F ion exchange column, carry out wash-out with distilled water, 0.3mol/L sodium chloride solution successively, and elutriant is collected in the test tube successively with partly collecting instrument automatically.Use 5: 1 sulfuric acid-phynol method of volume ratio respectively the elutriant in each test tube to be taken a sample and carry out color reaction, then with the spectrophotometric instrumentation 480nm OD of place value.Number be X-coordinate with pipe successively, the OD value is an ordinate zou, makes elution curve.According to elution curve, the elutriant at sodium chloride solution elution peak position merged and concentrate it, flowing water dialysis at least 36 hours, lyophilize, faint yellow solid.Faint yellow solid is redissolved in distilled water, with being splined on SephacrylS-300 molecular gel post successively with quadrat method and Sepharose CL-6B gel column carries out separation and purification, the elutriant that merges elution peak, detect its purity with HPLC then, if the HPLC elution curve is single symmetrical peak, just illustrate that this elutriant purity is up to standard; If not being single symmetrical peak, after then elutriant being concentrated, use the same method once more and be splined on Sephacryl S-300 molecular gel post and Sepharose CL-6B gel column carries out separation and purification, elution curve up to HPLC is single symmetrical peak, again this elutriant is concentrated postlyophilization, promptly get Cuttlebone polysaccharide CPS-1 of the present invention-1.
Experimentation on animals is that Cuttlebone polysaccharide CPS-1-1 is mixed with injection liquid, and abdominal injection has the obvious treatment effect in the mouse that suffers from ulcerative colitis.Therefore Cuttlebone polysaccharide CPS-1 of the present invention-1 can be used for preparing the medicine for the treatment of ulcerative colitis.
Description of drawings
Fig. 1 is the elution curve of Cuttlebone polysaccharide crude product through DEAE Sepharose F.F gel column
Fig. 2 is for presenting the HPLC elution curve of single symmetrical peak behind the elutriant purifying
Fig. 3 pair respectively organizes the content influence of EGF in the mice serum, PDGF, TNF-a for Cuttlebone polysaccharide CPS-1 of the present invention-1
Embodiment
Now in conjunction with the embodiments, the present invention is described in detail.
1. material
The Endoconcha Sepiae dry product is purchased in Zhoushan, Zhejiang
2. reagent
Phenol, sulfuric acid, chloroform, ethanol (analytical pure) (Shanghai chemical reagents corporation of Chinese Medicine group)
Sodium borohydride (NaBH
4) methyl-sulphoxide (DMSO), glucose and Dextran series dextran standard (SIGMA company)
3. instrument
H-S electric-heated thermostatic water bath (Dongtai electrical apparatus factory)
Rotary Evaporators B CHI 011 type (Switzerland B CHI company) BECKMAN COULTER J-251 type supercentrifuge (BECKMAN COULTER company)
DEAE Sepharose F.F filler, Sephacryl S-300 filler, the SepharoseCL-6B filler, part is collected instrument (Pharmacia Biotech company) automatically
High performance liquid phase HP1100 (Agilent company) KS-804, KS-805 post (Agilent company)
Embodiment 1: preparation Cuttlebone polysaccharide CPS-1-1
(1) preparation Cuttlebone polysaccharide crude product
The 5kg Endoconcha Sepiae was pulverized 3 minutes, add water 16L and boiled 10 hours, filtered through gauze while hot, residue repeats to extract twice, merges 3 times the water extract, is concentrated into 12L with Rotary Evaporators, and 5000rpm abandons precipitation after centrifugal 10 minutes.Supernatant liquor is that 4: 1 sevage method extraction removes albumen 3 times with the volume ratio of chloroform and propyl carbinol, does not contain protein to water, adds 3 times of volume ethanol alcohol precipitations then and spends the night.The centrifugal supernatant of abandoning will precipitate and redissolve in distilled water, and 5000rpm is centrifugal 10 minutes again, discards insolubles, and supernatant is with ethanol alcohol precipitation for the second time, the centrifugal supernatant that goes, and the precipitation dissolved in distilled water, flowing water was dialysed 36 hours.The dialyzate lyophilize obtains Cuttlebone polysaccharide crude product 10g.
(2) separation and purification, preparation Cuttlebone polysaccharide CPS-1-1
Get Cuttlebone polysaccharide crude product 1g, add distilled water 5ml, 80 ℃ of heating in water bath dissolved 30 minutes, and centrifuging and taking supernatant, insolubles add distilled water 5ml again, and 80 ℃ of heating in water bath dissolved the centrifuging and taking supernatant 30 minutes.Merge supernatant twice, be splined on DEAE Sepharose F.F post, use distilled water successively, 0.3mol/L sodium chloride solution wash-out, collect instrument with automatic part and collect elutriant in test tube, respectively color reaction is carried out in the elutriant in each test tube sampling with 5: 1 sulfuric acid-phynol method of volume ratio, with the spectrophotometric instrumentation 480nm OD of place value, with pipe number is X-coordinate, the OD value is an ordinate zou, draw elution curve and see Fig. 1, merge the elutriant at 0.3mol/L sodium chloride solution elution peak position and concentrate it, flowing water dialysis 48 hours according to elution curve, lyophilize gets faint yellow solid 0.78g.
Above-mentioned faint yellow solid 0.5g is dissolved in 3ml distilled water, is splined on SephacrylS-300 molecular gel post, the distilled water wash-out.Collecting instrument with method with automatic part and collect elutriant, carry out color reaction with 5: 1 sulfuric acid-phynol method of volume ratio, with the spectrophotometric instrumentation 480nm OD of place value, number is X-coordinate to manage, and the OD value is an ordinate zou, the drafting elution curve.The elutriant that merges the elution peak position according to elution curve, sampling detects its purity with HPLC, observe the elution curve that computer generates, see that it is not single symmetrical peak, illustrate that purity is not enough, after event concentrates elutriant, use the same method and be splined on Sepharose CL-6B gel column and carry out separation and purification, the elutriant that merges the elution peak position, detecting elution curve through HPLC is single symmetrical peak, sees Fig. 2, shows that this elutriant purity is good, so elutriant is concentrated postlyophilization, gets Cuttlebone polysaccharide CPS-1 of the present invention-1 100mg.
(3) detect Cuttlebone polysaccharide CPS-1-1 sugar degree and apparent molecular weight
1. with spectrophotometry Cuttlebone polysaccharide CPS-1-1 sugar degree:
(1) drafting of typical curve: be mixed with the aqueous solution that concentration is respectively 50 μ g/ml, 100 μ g/ml, 200 μ g/ml, 400 μ g/ml, 800 μ g/ml with standard glucose earlier, use 5: 1 sulfuric acid-phynol method of volume ratio to carry out color reaction respectively then, measure the OD of 480nm place value with spectrophotometry again, with sugared concentration is X-coordinate, the OD value is an ordinate zou, the drawing standard curve.
(2) sugar degree of mensuration Cuttlebone polysaccharide CPS-1-1: the solution that Cuttlebone polysaccharide CPS-1-1 is mixed with 1mg/ml concentration, carry out color reaction with 5: 1 sulfuric acid-phynol method of volume ratio, with the spectrophotometric determination 480nm OD of place value, contrast glucose typical curve gets corresponding sugared concentration then.The sugar degree that calculating can get Cuttlebone polysaccharide CPS-1-1 is 96.6%.
2. measure the CPS-1 apparent molecular weight with the HPLC method:
(1) drafting of typical curve: Dextran T series standard molecular weight is respectively 5000,10000,100000,400000,1000000 dextran is mixed with the 2mg/ml aqueous solution, uses behind the 0.45 μ m pin type filtering with microporous membrane successively sample introduction to measure retention time in HPLC respectively.The chromatographic condition of HPLC is: separator column is KS-805, the KS-804 columns in series; Moving phase is water; It is 254nm that the VWD detector detects wavelength; Differential detector (RID) detection cell temperature is 40 ℃; Be 40 minutes analysis time; Sampling volume is 25 μ l.With the retention time is X-coordinate, and the molecular weight of standard dextran is an ordinate zou, makes typical curve.
(2) measure the apparent molecular weight of Cuttlebone polysaccharide CPS-1-1: Cuttlebone polysaccharide CPS-1-1 is mixed with the aqueous solution of 2mg/ml, with sample introduction behind the 0.45 μ m pin type filtering with microporous membrane in HPLC, the mensuration retention time.The chromatographic condition of HPLC is the same.According to retention time, contrast above-mentioned typical curve, its apparent molecular weight is 1.1 * 10
5Dalton.
Embodiment 2: preparation Cuttlebone polysaccharide CPS-1-1
The 5kg Endoconcha Sepiae was pulverized 5 minutes, add water 12L and boiled 9 hours, filtered through gauze while hot, residue repeats to extract twice, merges 3 times the water extract, is concentrated into 12L with Rotary Evaporators, and 5000rpm abandons precipitation after centrifugal 10 minutes.Supernatant liquor is that 4: 1 sevage method extraction removes albumen 3 times with the volume ratio of chloroform and propyl carbinol, does not contain protein to water, and the ethanol alcohol precipitation that adds 3 times of volumes then spends the night.The centrifugal supernatant of abandoning will precipitate and redissolve in distilled water, and 5000rpm is centrifugal 10 minutes again, discards insolubles, and supernatant is with ethanol alcohol precipitation for the second time, the centrifugal supernatant that goes, and the precipitation dissolved in distilled water, flowing water was dialysed 40 hours.The dialyzate lyophilize obtains Cuttlebone polysaccharide crude product 9g.
The sugar degree of separation and purification and mensuration Cuttlebone polysaccharide CPS-1-1 and the method for apparent molecular weight are with embodiment 1.The sugar degree that calculates Cuttlebone polysaccharide CPS-1-1 is 95.7%, and its apparent molecular weight is 1.1 * 10
5Dalton.
Embodiment 3: preparation Cuttlebone polysaccharide CPS-1-1
The 5kg Endoconcha Sepiae was pulverized 8 minutes, add water 14L and boiled 14 hours, filtered through gauze while hot, residue repeats to extract twice, merges 3 times the water extract, is concentrated into 12L with Rotary Evaporators, and 5000rpm abandons precipitation after centrifugal 10 minutes.Supernatant liquor is that 4: 1 sevage method extraction removes albumen 3 times with the volume ratio of chloroform and propyl carbinol, does not contain protein to water, and the ethanol alcohol precipitation that adds 3 times of volumes then spends the night.The centrifugal supernatant of abandoning will precipitate and redissolve in distilled water, and 5000rpm is centrifugal 10 minutes again, discards insolubles, and supernatant is with ethanol alcohol precipitation for the second time, the centrifugal supernatant that goes, and the precipitation dissolved in distilled water, flowing water was dialysed 48 hours.The dialyzate lyophilize obtains Cuttlebone polysaccharide crude product 12g.
The sugar degree of separation and purification and mensuration Cuttlebone polysaccharide CPS-1-1 and the method for apparent molecular weight are with embodiment 1.The sugar degree that calculates Cuttlebone polysaccharide CPS-1-1 is 98.5%, and its apparent molecular weight is 1.1 * 10
5Dalton.
Embodiment 4: the therapeutic test of Cuttlebone polysaccharide CPS-1-1 pair mouse ulcerative colitis
1 material
Cuttlebone polysaccharide CPS-1-1 is made by embodiment 1.
The BALA/C mouse is purchased the Experimental Animal Center confession in The 2nd Army Medical College, and in 9 ages in week, body weight is 25 ± 2.2g.
2 reagent
Dextran sulfate sodium (DSS) (Sigma company, relative molecular mass 5000)
Mouse EGF ELISA detection kit (Shanghai Xi Tang biotech firm)
Mouse TNF-a ELISA detection kit (Shanghai Xi Tang biotech firm)
Mouse PDGF ELISA detection kit (Shanghai Xi Tang biotech firm)
3 instruments
BIO-RAD 680 type microplate reader (BIO-RAD company)
BECKMAN table model high speed centrifuge (BECKMAN company)
4. experimental technique and result
4.1 the foundation of mouse ulcerative colitis treatment model
30 BALA/C mouse are divided into 5 groups at random, 6 every group, are respectively blank group, positive controls, negative control group, CPS-1 low concentration group and CPS-1 high density group.Except that the blank group is normal raise, all the other every group of mouse are freely drunk 5% the DSS aqueous solution and induce ulcerative colitis, drink totally 7 days, promptly build up acute ulcer colitis model.From drinking the 5%DSS aqueous solution the 3rd day, the mouse of positive controls is injected the 0.1ml ulcerative colitis medicine compound sulfonamide aqueous solution every day, and concentration is 2.5mg/ml; The CPS-1 low concentration group is injected the Cuttlebone polysaccharide CPS-1 that 0.1ml concentration is 2mg/ml-1 aqueous solution every day; CPS-1 high density group is injected the Cuttlebone polysaccharide CPS-1 that 0.1ml concentration is 5mg/ml-1 aqueous solution every day; Negative control group is then injected 0.1ml physiological saline every day.
4.2 the animal of experiment mice and cell levels are observed
4.2.1 the observation of experiment mice generalized case
Experiment mice rose drinking 5%DSS solution on the 2nd day, made fecal occult blood test every day respectively, and observed diarrhoea and naked eyes bloody stool situation.The variation of experiment mice body weight before and after the record modeling, each cell mean sees Table 1:
| Group | Body weight (g) before the model | Body weight behind the model (g) |
| Blank positive control negative control CPS-1 (lower concentration) CPS-1 (high density) | 25.21±0.56 24.86±0.37 25.62±0.13 24.79±0.59 25.53±0.22 | 35.15±0.23 30.84±0.85 24.47±0.63 23.57±0.76 33.11±0.74 |
Mouse body weight change before and after table 1 modeling (mean body weight ± SD)
By table 1 as seen, body weight and the blank winding of Cuttlebone polysaccharide CPS-1-1 high density treatment group mouse are near, and be slightly heavier than positive controls, apparently higher than negative control group, illustrates that the ulcerative colitis of Cuttlebone polysaccharide CPS-1-1 pair experiment mice has therapeutic action.
4.2.2 the experiment mice stereomicroscope is checked
Drink the 5%DSS aqueous solution after 7 days, the 8th day with the execution of every group of mouse usefulness cervical vertebra dislocation method.Open the abdominal cavity, isolate total colectomy, place D-hank ' s liquid.Cut off the intestines wall along the mesentery facies posterior hepatis, intestinal contents is rinsed the back well and on filter paper the intestines wall is launched, and the rearmounted stereomicroscope of Toluidine blue staining is observed the change of intestinal mucosa down.Extensively congested, the oedema of the intestinal mucosa of visible negative control group experiment mice under the stereomicroscope, there is erosion, hemorrhage the part, and can find tangible ulcer kitchen range, and is especially remarkable with the lower semisection colon.The intestinal mucosa of positive controls, the CPS-1 low dose group experiment mice degree of festering is not high, and significantly the ulcer kitchen range does not occur in the group on one's body every animal.And the intestinal mucosa of blank group and CPS-1 high dose group experiment mice slightly festers, and has tangible ulcer kitchen range to occur once in a while.This shows that the ulcerative colitis of Cuttlebone polysaccharide CPS-1-1 pair experiment mice has the obvious treatment effect, and Cuttlebone polysaccharide CPS-1-1 concentration is big more, result of treatment is good more.
4.4 the detection of cytokine content relatively in the experiment mice blood
4.4.1 the preparation of experiment mice blood specimen: every group of experiment mice carries out eye socket and gets blood 0.2ml, adds the heparin sodium anti-freezing, and 5000rpm abandons precipitation and obtains serum after centrifugal 15 minutes.Marking serial numbers is standby respectively.
4.4.2 EGF, TNF-a, PDGF Determination on content in the experiment mice blood: use mouse ELISA test kit that the EGF in the serum sample, TNF-a, PDGF content are carried out detection by quantitative.The typical curve that microplate reader reads behind the plate to provide in the contrast agents box as a result calculates the content of every kind of cytokine respectively, and carries out errot analysis and significant difference analysis with statistical method.At last the result is seen Fig. 3 with the mapping of EXCEL software.
As seen from Figure 3, Cuttlebone polysaccharide CPS-1-1 can significantly improve the expression level of EGF and PDGF in the experiment mice blood, and the expression level of TNF-a descends to some extent.The increase that EGF expresses helps the healing of chronic ulcer tissue, and PDGF plays active effect to the reparation of chronic ulcer tissue equally, and the expression level of TNF-a is the significant cytokine of chronic ulcer tissue inflammation degree always.Therefore, Cuttlebone polysaccharide CPS-1-1 can play active effect to the healing of chronic ulcer tissue, promotes the expression of EGF and PDGF on the one hand, accelerates the cytothesis process, on the other hand, suppresses the expression of TNF-a, thus amelioration of inflammation.
Preparation method of the present invention is simple, above-mentioned experiment confirm the ulcerative colitis of Cuttlebone polysaccharide CPS-1-1 pair experiment mice have significant therapeutic action, so Cuttlebone polysaccharide CPS-1 of the present invention-1 can be used for preparing the medicine for the treatment of ulcerative colitis.
Claims (3)
1. the preparation method of Cuttlebone polysaccharide CPS-1-1, step is as follows:
(1) preparation water extract solution: the Endoconcha Sepiae solid is ground into powder, adds water boil 8-10 hour, suction filtration obtains filtrate while hot, filtrate is concentrated the centrifugal precipitation of abandoning; Supernatant liquor is that 4: 1 sevage method extraction repeatedly removes albumen with the volume ratio of chloroform and propyl carbinol, does not contain protein to water, obtains water extract solution;
(2) preparation Cuttlebone polysaccharide crude product: said extracted thing solution is added ethanol sedimentation spend the night, the centrifugal supernatant of abandoning will precipitate and redissolve in distilled water, and the centrifugal insolubles that goes is with supernatant alcohol precipitation once more, the centrifugal supernatant that goes; The dissolving of precipitation adding distil water, flowing water dialysis at least 36 hours; With the dialyzate lyophilize, get the Cuttlebone polysaccharide crude product;
(3) separation and purification, preparation Cuttlebone polysaccharide CPS-1-1: will be dissolved in distilled water in the hot water bath of gained Cuttlebone polysaccharide crude product, centrifugal collection supernatant, insolubles is dissolved in distilled water in the hot water bath once more, the centrifugal precipitation of abandoning merges twice supernatant liquor, is splined on DEAE SepharoseF.F ion exchange column, carry out wash-out with distilled water, 0.3mol/L sodium chloride solution successively, and be collected in elutriant in the test tube successively with partly collecting instrument automatically; Use 5: 1 sulfuric acid-phynol method of volume ratio respectively the elutriant in each test tube to be taken a sample and carry out color reaction, then with the spectrophotometric instrumentation 480nm OD of place value; Number be X-coordinate with pipe successively, the OD value is an ordinate zou, makes elution curve; According to elution curve, the elutriant at sodium chloride solution elution peak position merged and concentrate it, flowing water dialysis at least 36 hours, lyophilize, faint yellow solid; Faint yellow solid is redissolved in distilled water, with being splined on SephacrylS-300 molecular gel post successively with quadrat method and Sepharose CL-6B gel column carries out separation and purification, the elutriant that merges the elution peak position, take a sample then and detect purity with HPLC, observe the elution curve that computer generates, if single symmetrical peak, show that purity is good, if not single symmetrical peak, after then elutriant being concentrated, carrying out separation and purification with Sephacryl S-300 molecular gel post and SepharoseCL-6B gel column once more, is single symmetrical peak up to the elution curve of HPLC; Again this elutriant is concentrated postlyophilization, promptly get Cuttlebone polysaccharide CPS-1 of the present invention-1.
2. the Cuttlebone polysaccharide CPS-1-1 of the described method of claim 1 preparation.
3. the application of claim 2 described Cuttlebone polysaccharide CPS-1-1 in preparation treatment ulcerative colitis medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101100826A CN100362022C (en) | 2005-11-07 | 2005-11-07 | Cuttlebone polysaccharide CPS-1 and its preparation and use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101100826A CN100362022C (en) | 2005-11-07 | 2005-11-07 | Cuttlebone polysaccharide CPS-1 and its preparation and use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1837243A CN1837243A (en) | 2006-09-27 |
| CN100362022C true CN100362022C (en) | 2008-01-16 |
Family
ID=37014767
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005101100826A Expired - Fee Related CN100362022C (en) | 2005-11-07 | 2005-11-07 | Cuttlebone polysaccharide CPS-1 and its preparation and use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100362022C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401815B (en) * | 2008-11-18 | 2011-05-04 | 中国人民解放军第二军医大学 | Cuttlebone polysaccharide extract CBP-S and uses thereof |
| CN104119428B (en) * | 2013-04-24 | 2017-09-12 | 启东盖天力药业有限公司 | A kind of Huai Er albumen and its production and use |
| CN103755820A (en) * | 2013-10-25 | 2014-04-30 | 南通瑞普埃尔生物工程有限公司 | Cuttlebone polysaccharide for treating empyrosis and preparing process thereof |
| CN103969384B (en) * | 2014-05-06 | 2016-04-20 | 济南康众医药科技开发有限公司 | A kind of content assaying method of blood clam polysaccharide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04138169A (en) * | 1990-09-28 | 1992-05-12 | Tottori Univ | Living body internal filler |
| CN1076859A (en) * | 1992-03-30 | 1993-10-06 | 河南省新乡市前卫制药厂 | The extraction of the flat effective ingredient of the cancer that disappears and dosage form preparation |
| JPH0840915A (en) * | 1994-07-27 | 1996-02-13 | San Five Kk | Anti-inflammatory agent |
| CN1419566A (en) * | 2000-03-30 | 2003-05-21 | 阿默森生物科学有限公司 | A method of production of LgG |
-
2005
- 2005-11-07 CN CNB2005101100826A patent/CN100362022C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04138169A (en) * | 1990-09-28 | 1992-05-12 | Tottori Univ | Living body internal filler |
| CN1076859A (en) * | 1992-03-30 | 1993-10-06 | 河南省新乡市前卫制药厂 | The extraction of the flat effective ingredient of the cancer that disappears and dosage form preparation |
| JPH0840915A (en) * | 1994-07-27 | 1996-02-13 | San Five Kk | Anti-inflammatory agent |
| CN1419566A (en) * | 2000-03-30 | 2003-05-21 | 阿默森生物科学有限公司 | A method of production of LgG |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1837243A (en) | 2006-09-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102600219B (en) | Total flavone extract of abelmoschus manihot and preparing method of total flavone extract | |
| Dandjesso et al. | Phytochemistry and hemostatic properties of some medicinal plants sold as anti-hemorrhagic in Cotonou markets (Benin) | |
| CN103222988B (en) | A kind of American-cockroach-extract and its preparation method and application | |
| CN103435580B (en) | Lingzhiol A and application of lingzhiol A in drug production and foods | |
| CN102617745B (en) | Preparation method and blood sugar lowering function of Ganoderma lucidum karst polysaccharide F31 | |
| CN104710538B (en) | A kind of sanchi flower arabogalactan and its production and use | |
| CN102652792B (en) | Anti-cancer application of composition containing garlic oil, garlic total polysaccharide and garlic total saponin | |
| KR20040074614A (en) | Medicinal preparation containing phenylethanoid glycosides extracted from herbaceous plant, cistanche tubulosa(schenk.)wight, process of making the same, and uses of the same | |
| CN105037577B (en) | Procoagulant blackberry seed polysaccharide, and extraction separation method and application thereof | |
| CN100362022C (en) | Cuttlebone polysaccharide CPS-1 and its preparation and use | |
| CN101829224B (en) | Traditional Chinese medicine composition preparation and preparation method and quality control method | |
| CN104107187B (en) | The application in treatment diabetes medicament of a kind of Herba Apocyni veneti acidic polysaccharose | |
| CN101028317B (en) | Use of hypericum japonicum in preparation of medicine against nephritis and renal insufficiency | |
| CN104910291B (en) | A kind of jackfruit leaf polyose and its preparation method and application | |
| CN107149631A (en) | A kind of method for separating and preparing of Kwangtung purple beautyberry extract and application thereof | |
| CN101468104B (en) | Chinese medicinal compound preparation for treating osteoporosis and method for preparing the same | |
| CN101948549B (en) | Sulphating modification method of gynostemma pentaphylla polysaccharide | |
| CN101695511A (en) | Pomegranate rind extract and production method and application thereof | |
| CN102319420B (en) | The application of turtle peptide in pharmacy | |
| CN110568120B (en) | Loranthus parasiticus quality control method based on double-substance components | |
| CN115998781A (en) | Application of birch juice separating liquid in treating gout | |
| CN102274262B (en) | Application of Scorzonera total extract in preparation of medicaments for treating hepatitis or protecting liver | |
| CN103897072A (en) | Bock greenbrier rhizome polysaccharide extract and preparation method thereof | |
| CN106176775B (en) | Forsythiaside B and poliumoside compatible composition and its application | |
| CN107998324A (en) | A kind of Chinese medicine preparation for being used to reduce chronic renal albuminuria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080116 Termination date: 20101107 |