CN104263790B - A kind of saline cistanche polypeptide extract, preparation method and its usage - Google Patents
A kind of saline cistanche polypeptide extract, preparation method and its usage Download PDFInfo
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Abstract
The invention provides a kind of method that saline cistanche polypeptide extract is extracted from saline cistanche, and its saline cistanche polypeptide extract and the purposes of the saline cistanche polypeptide extract that thus method is obtained.The present invention utilizes saline cistanche slag, drying is crushed, again by alkali extraction-acid precipitation processing, enzymolysis, ultrafiltration purification, the molecular weight finally given is the saline cistanche polypeptide of 500 10000 dalton, and the effect suffered from diarrhoea caused by saline cistanche polypeptide therapeutic cancer and cancer chemotherapy is studied.The present invention improves the utilization rate of Precious, Rare, Endangered medicinal material Herba Cistanches, it is set to produce higher economic value, provide a kind of high-efficiency low-toxicity for vast tumor patient simultaneously, it is cheap, it is easy to popularization available for mitigation cancer chemotherapy induced diarrhea and play the medicine of chemotherapeutic sensitivity.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to a kind of side that saline cistanche polypeptide extract is extracted from saline cistanche
Method, and its saline cistanche polypeptide extract and the purposes of the saline cistanche polypeptide extract that thus method is obtained.
Background technology
Saline cistanche (Cistanche deserticola Y.C.Ma) is Orobanchaceae (Orobanchaeceae) Cistanche deserticola
(Cistanche) fleshy stem of plant saline cistanche dry zone scale leaf, the smart bamboo shoot of fork name, vertical Rong, dried meat floss Rong, smart, big rue, have " husky
The good reputation of unconcerned ginseng ", begins to be recorded in《Sheng Nong's herbal classic》, top grade is classified as, is one of conventional Chinese medicine, with tonifying kidney and strengthening yang, replenishing essence
The effects such as mends marrow, enriching the blood to restore normal menstruation, happy color macrobiosis.About 18 kinds in this platymiscium whole world, respectively China's 1 mutation of about 4 kinds, meat
Desert cistanche, halophilous herbage, Cistanche tubulosa, spend salt saline cistanche and husky desert cistanche in vain, remove sand outside desert cistanche, other 4 kinds of main products are in China
Each provinces and regions in northwest.
Early 1980s Japanese scholars begin one's study the chemical composition of saline cistanche.To the late 1980s, state
Interior scholar begin one's study China's output saline cistanche chemical composition.With separation is extracted and detection technique is developed rapidly,
A variety of chemical compositions such as isolated benzyl carbinol glycoside, iridoids, volatile ingredient, lignanoids, polysaccharide, alkaloid.
Modern pharmacology research also demonstrates that saline cistanche in addition to tonifying kidney and strengthening yang, functions of loosening bowel relieving constipation, also with anti-aging
Effect, and without find adverse reaction.Saline cistanche is widely used in each similar drug and health products in recent years.
The traditional preliminary working method of Herba Cistanches has drying method, salt marsh method, stores sth. in a cellar method etc., and these methods are time-consuming, by certainly
The influence of right condition, and quality is unstable, has a strong impact on the utilization rate and curative effect of medicinal material；Also, traditional preliminary working method oneself
The need for a large amount of medicinal material preliminary working in the place of production far can not be met.Therefore, the utilization rate of Precious, Rare, Endangered medicinal material Herba Cistanches how is improved,
It is produced higher economic value, be witch's key issue to be solved.
Chinese patent CN102349650A discloses a kind of method for improving dissolution rate of active ingredients of Cistanche salsa, invention master
To cross Rapid Cleaning by fresh herba cistanches, then cutting, citric acid solution immersion, after microwave deactivating enzyme, add amylase and
Pectase is digested, and then adds filter aid, and enzymolysis liquid is separated.The technical problem that the patent is solved is to improve juice
Rate, makes crushing juice rate bring up to 65% by 54%, so as to improve the dissolution rate of saline cistanche active ingredient, the patent does not provide this hair
The technical inspiration of bright saline cistanche polypeptide and preparation method thereof.
This patent uses Takla Makan Desert high-quality saline cistanche for raw material, and peeling processing, gained are carried out to its fresh medicine material
Saline cistanche skin be dried and extracted again using ethanol, traditional chemical composition can be obtained；Juice extractor is used to pulp
Squeezed the juice, obtain water extract；Saline cistanche slag obtained by after to squeezing the juice carries out drying and processing, is ground into powder, then by alkali
The molten heavy method processing of acid, enzymolysis, ultrafiltration purification, the molecular weight finally given is the polypeptide of 500-10000 dalton, and is treated
The research of diarrhoea effect caused by cancer and cancer chemotherapy, and with Chinese medicine composition disclosed in Chinese patent CN103585546A
The research that compatibility is used.
Current cancer turns into a big killer of facing mankind, and the death rate is only second to angiocardiopathy, and on year by year
The trend risen.Tumor patient carries out chemotherapy, although have certain curative effect, but after chemotherapy most of patient occur it is different degrees of
Side effect, wherein chemotherapeutic be alone or the joint toxicity, being called of diarrhoea after chemotherapy of tumors such as can cause patient to suffer from diarrhoea
Treat induced diarrhea.Recently as chemotherapeutics such as 5- fluorouracils, taxanes, HCPT, Irinotecan, uncommon
Luo Da's etc. is emerged and popularization and application, and the incidence of Chemotherapy-induced Diarrhea is improved year by year, and if without active treatment, easily
Cause patient's dehydration, malnutrition, serum electrolyte disorder, acid-base imbalance etc., very big not only is come to the recovery band of body
It is unfavorable, also bring certain difficulty to periodic chemotherapy, therefore tumour is turned into effective treatment of Chemotherapy-induced Diarrhea
The important component of patient's wholistic therapy.
It there is no confrontation cancer chemotherapy that there is alleviating diarrhoea and the Chinese patent drug of synergistic effect, Chinese patent at present
Chinese medicine composition disclosed in CN103585546A is indefinite in the mechanism of action suffered from diarrhoea caused by treating cancer chemotherapy, and drug effect is failed to understand
It is aobvious, thus under the auxiliary of this Chinese medicine composition, research and develop a kind of high-efficiency low-toxicity, it is cheap, it is easy to can be used for of popularization, mitigates
Cancer chemotherapy induced diarrhea simultaneously plays the pharmaceutical composition of chemotherapeutic sensitivity and seems and be even more important.
The content of the invention
In order to solve defect present in prior art, it is an object of the invention to provide saline cistanche polypeptide extract, system
Preparation Method and application thereof.The method that saline cistanche polypeptide extract is prepared in a kind of powder from saline cistanche that the present invention is provided, including with
Lower step：
（1）It is 8-10, the extraction time 40-60min at 45-55 DEG C to add NaOH solution to pH value toward saline cistanche powder, so
Centrifugal treating, produces saline cistanche albumen afterwards；
（2）Use step（1）Obtained saline cistanche albumen configuration concentration of substrate 5-9% saline cistanche albumino reaction system, is adjusted
The pH value of saline cistanche albumino reaction system is saved to 6.0-7.0,60-70 DEG C of temperature is controlled, adds enzymolysis of the enzyme bottom than 3.0-4.0%
Protease, constant temperature stirring enzymolysis, enzymolysis time is 3.0-4.0h, produces enzymolysis liquid, and described enzymolysis protein enzyme is basic protein
At least one of enzyme, papain, neutral proteinase, pectase and amylase；
（3）To step（2）Obtained enzymolysis liquid progress 12-17min's boils destroy the enzyme treatment, and 2 mol/L are used after cooling
HCL solution be adjusted to pH value to neutrality, be then centrifuged for remove precipitation, take supernatant, supernatant contains saline cistanche protein peptides；
（4）By step（3）The resulting supernatant containing saline cistanche protein peptides, is 40-50 DEG C in temperature, pH value is
7.0-9.0, pressure is under 0.7-0.9MPa, selection 500Da and l0000Da milipore filter is to 500- in saline cistanche protein peptides
The saline cistanche polypeptide of 10000Da molecular weight is separated, and obtains the leaching for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C
Cream, as saline cistanche polypeptide extract.
Further, the side of saline cistanche polypeptide extract is prepared during the present invention improves a kind of optimal powder from saline cistanche
Method, is comprised the steps of：
（1）It is 9, the extraction time 50min at 50 DEG C to add NaOH solution to pH value toward saline cistanche powder, is then centrifuged for place
Reason, produces saline cistanche albumen；
（2）Use step（1）Obtained saline cistanche albumen configures the saline cistanche albumino reaction system of concentration of substrate 7%, regulation
The pH value of saline cistanche albumino reaction system controls 65 DEG C of temperature to 6.5, adds enzymolysis protein enzyme of the enzyme bottom than 3.5%, and constant temperature is stirred
Enzymolysis is mixed, enzymolysis time is 3.5h, produce enzymolysis liquid, described enzymolysis protein enzyme is papain；
（3）To step（2）Obtained enzymolysis liquid progress 15min's boils destroy the enzyme treatment, uses 2 mol/L's after cooling
HCL solution is adjusted to pH value to neutrality, is then centrifuged for removing precipitation, takes supernatant, supernatant contains saline cistanche protein peptides；
（4）By step（3）The resulting supernatant containing saline cistanche protein peptides, is 45 DEG C in temperature, pH value is 8.0,
Pressure is under 0.8MPa, selection 500Da and l0000Da milipore filter is to 500-10000Da molecular weight in saline cistanche protein peptides
Saline cistanche polypeptide is separated, and obtains the medicinal extract for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C, and as saline cistanche is more
Peptide extract.
By detection, the content of the active peptides in the saline cistanche polypeptide extraction that the above method is obtained is 99%, active peptides
Molecular weight ranges between 500-10000 dalton.
Second object of the present invention is to provide a kind of saline cistanche polypeptide obtained according to the method described above from saline cistanche powder and carried
Take in thing, saline cistanche polypeptide extract contain saline cistanche polypeptide 99%, and saline cistanche polypeptide molecular weight in 500-10000 roads
Between you pause.
Third object of the present invention is to provide saline cistanche polypeptide extract for preparing in treating cancer medicine
Purposes.
Fourth object of the present invention is that provide saline cistanche polypeptide extract causes for preparing treating cancer chemotherapy
Diarrhoea medicine in purposes.
Further, described medicine is the composition being made up of the raw material of following parts by weight：Saline cistanche polypeptide extract
5-16 parts, 7-9 parts of hawthorn, 5-8 parts of red yeast rice, 6-14 parts of radish seed, 3-9 parts of the coptis, 5-9 parts of gynostemma pentaphylla, 5-12 parts of sweet osmanthus, pseudo-ginseng
5-14 parts, 8-12 parts of the red sage root, 9-15 parts of the Radix Astragali, 6-14 parts of fructus amomi, 4-6 parts of radix scutellariae, 6-12 parts of forrest silkvine stem or root, 5-12 parts of japanese rose root,
6-10 parts of 5-9 parts of rhizoma zingiberis and Piper longum.
Further, the preparation method of described medicine includes following step：The above-mentioned medicinal material of recipe quantity is taken to be broken into carefully
Except powder, saline cistanche polypeptide extract, the ethanol solution that volumetric concentration is 75-90% is added according to 5-8 times of coarse powder gross weight,
Refluxing extraction three times, return time is 2-4h, filtering, adds and is concentrated to give at saline cistanche water extract, 60 DEG C in the filtrate of gained
To the medicinal extract that relative density is 1.20, that is, obtain drug extract of the present invention.
Further, described medicine is powder, aqua, tablet or capsule.
In a word, the present invention compared with prior art, with technical advantage prominent as follows：
（1）The present invention uses saline cistanche for raw material, and peeling processing is carried out to its fresh medicine material, and the saline cistanche skin of gained is done
It is dry to be extracted again using ethanol, traditional chemical composition can be obtained；Pulp is squeezed the juice using juice extractor, water extraction is obtained
Take thing；Saline cistanche slag obtained by after to squeezing the juice carries out drying and processing, is ground into powder, then handle, digest by alkali extraction-acid precipitation,
Ultrafiltration purification, the molecular weight finally given is the polypeptide of 500-10000 dalton, has prepared great drug value
Saline cistanche polypeptide extract, improves the utilization rate of Precious, Rare, Endangered medicinal material Herba Cistanches, it is produced higher economic value.
（2）The present invention prepares biologically active peptide using enzymatic isolation method, and imitates the normal digestion conditions in human body alimentary canal will
Saline cistanche Protein cleavage has larger advantage compared with inorganic acid, basic hydrolysis into specific proteolytie fragrnent, mild condition, it is to avoid water
Problem of environmental pollution in solution preocess.
（3）The present invention makes gained polypeptide using the polypeptide fragment in ultrafiltration means directional collecting 500-10000 dalton
Purity in the molecular weight ranges of this in composition is up to 99%, and process stabilizing, and reappearance is high.
（4）The present invention also proves that saline cistanche polypeptide extract of the present invention and medicine of the present invention can be notable by effect experiment
Suppress the growth of cancer cell, show that the medicine of the present invention has significant therapeutic effect to cancer.And saline cistanche polypeptide extract
Determined curative effect when being used for the treatment suffered from diarrhoea caused by cancer and cancer chemotherapy with corresponding medicine, adverse reaction and side effect are equal
Substantially reduction, it has significant synergy with the Chinese medicine suffered from diarrhoea caused by existing treating cancer chemotherapy in terms of antidiarrheal, and
And the symptom of diarrhea of cancer chemotherapy initiation can be greatly reduced, improve cancer patient and complied with for the acceptance of chemotherapy and medication
Property.
Embodiment
The present invention is further described below by way of specific embodiment, but the present invention is not limited only to following examples.At this
In the range of invention or do not departing from present disclosure, in spirit and scope, saline cistanche polypeptide of the present invention is being carried
Take thing, preparation method and purposes only to carry out change nominally, replace, be apparent to those skilled in the art
, they are considered as being included within the scope of the present invention.
The preparation of Part I, saline cistanche polypeptide extract
A kind of method that saline cistanche polypeptide extract is prepared in embodiment 1, powder from saline cistanche：
（1）It is 10, the extraction time 60min at 45 DEG C to add NaOH solution to pH value toward saline cistanche powder, is then centrifuged for place
Reason, produces saline cistanche albumen；
（2）Use step（1）Obtained saline cistanche albumen configures the saline cistanche albumino reaction system of concentration of substrate 5%, regulation
The pH value of saline cistanche albumino reaction system controls temperature 70 C to 6.0, adds enzymolysis protein enzyme of the enzyme bottom than 4.0%, and constant temperature is stirred
Enzymolysis is mixed, enzymolysis time is 4.0h, produce enzymolysis liquid, described enzymolysis protein enzyme is papain；
（3）To step（2）Obtained enzymolysis liquid progress 17min's boils destroy the enzyme treatment, uses 2 mol/L's after cooling
HCL solution is adjusted to pH value to neutrality, is then centrifuged for removing precipitation, takes supernatant, supernatant contains saline cistanche protein peptides；
（4）By step（3）The resulting supernatant containing saline cistanche protein peptides, is 40 DEG C in temperature, pH value is 7.0,
Pressure is under 0.7MPa, selection 500Da and l0000Da milipore filter is to 500-10000Da molecular weight in saline cistanche protein peptides
Saline cistanche polypeptide is separated, and obtains the medicinal extract for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C, and as saline cistanche is more
Peptide extract.
The content of active peptides in the saline cistanche polypeptide extract obtained by detection, the above method is 92%.
A kind of method that saline cistanche polypeptide extract is prepared in embodiment 2, powder from saline cistanche：
（1）It is 8, the extraction time 40min at 55 DEG C to add NaOH solution to pH value toward saline cistanche powder, is then centrifuged for place
Reason, produces saline cistanche albumen；
（2）Use step（1）Obtained saline cistanche albumen configures the saline cistanche albumino reaction system of concentration of substrate 9%, regulation
The pH value of saline cistanche albumino reaction system controls temperature 60 C to 7.0, adds enzymolysis protein enzyme of the enzyme bottom than 3.0%, and constant temperature is stirred
Enzymolysis is mixed, enzymolysis time is 3.0h, produce enzymolysis liquid, described enzymolysis protein enzyme is papain；
（3）To step（2）Obtained enzymolysis liquid progress 15min's boils destroy the enzyme treatment, uses 2 mol/L's after cooling
HCL solution is adjusted to pH value to neutrality, is then centrifuged for removing precipitation, takes supernatant, supernatant contains saline cistanche protein peptides；
（4）By step（3）The resulting supernatant containing saline cistanche protein peptides, is 50 DEG C in temperature, pH value is 9.0,
Pressure is under 0.9MPa, selection 500Da and l0000Da milipore filter is to 500-10000Da molecular weight in saline cistanche protein peptides
Saline cistanche polypeptide is separated, and obtains the medicinal extract for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C, and as saline cistanche is more
Peptide extract.
The content of active peptides in the saline cistanche polypeptide extract obtained by detection, the above method is 93%.
A kind of method that saline cistanche polypeptide extract is prepared in embodiment 3, powder from saline cistanche：
（1）It is 9, the extraction time 50min at 50 DEG C to add NaOH solution to pH value toward saline cistanche powder, is then centrifuged for place
Reason, produces saline cistanche albumen；
（2）Use step（1）Obtained saline cistanche albumen configures the saline cistanche albumino reaction system of concentration of substrate 7%, regulation
The pH value of saline cistanche albumino reaction system controls 65 DEG C of temperature to 6.5, adds enzymolysis protein enzyme of the enzyme bottom than 3.5%, and constant temperature is stirred
Enzymolysis is mixed, enzymolysis time is 3.5h, produce enzymolysis liquid, described enzymolysis protein enzyme is papain；
（3）To step（2）Obtained enzymolysis liquid progress 15min's boils destroy the enzyme treatment, uses 2 mol/L's after cooling
HCL solution is adjusted to pH value to neutrality, is then centrifuged for removing precipitation, takes supernatant, supernatant contains saline cistanche protein peptides；
（4）By step（3）The resulting supernatant containing saline cistanche protein peptides, is 45 DEG C in temperature, pH value is 8.0,
Pressure is under 0.8MPa, selection 500Da and l0000Da milipore filter is to 500-10000Da molecular weight in saline cistanche protein peptides
Saline cistanche polypeptide is separated, and obtains the medicinal extract for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C, and as saline cistanche is more
Peptide extract.
The content of active peptides in the saline cistanche polypeptide extract obtained by detection, the above method is 99%.
Part II, the pharmaceutical preparation of the invention containing saline cistanche polypeptide extract
Embodiment 4, medicine of the present invention and preparation technology：
The parts by weight of drug component：5 parts of saline cistanche polypeptide extract, 7 parts of hawthorn, 5 parts of red yeast rice, 6 parts of radish seed, the coptis 3
Part, 5 parts of gynostemma pentaphylla, 5 parts of sweet osmanthus, 5 parts of pseudo-ginseng, 8 parts of the red sage root, 9 parts of the Radix Astragali, 6 parts of fructus amomi, 4 parts of radix scutellariae, 6 parts of forrest silkvine stem or root, japanese rose root
6 parts of 5 parts, 5 parts of rhizoma zingiberis and Piper longum.The above-mentioned medicinal material of recipe quantity is taken to be broken into except fine powder, saline cistanche polypeptide extract, according to coarse powder
The ethanol solution that 5 times of addition volumetric concentrations of gross weight are 75%, refluxing extraction three times, return time is 3h, filtering, gained
The medicinal extract for being concentrated to give that relative density is 1.20 at saline cistanche water extract, 60 DEG C is added in filtrate, that is, obtains medicine of the present invention
Medicinal extract.Clinically conventional Chinese medicine pharmaceutical dosage form, such as tablet or capsule can be prepared by preparation process thereof commonly used in the art
Agent.
Embodiment 5, medicine of the present invention and preparation technology：
The parts by weight of drug component：16 parts of saline cistanche polypeptide extract, 9 parts of hawthorn, 8 parts of red yeast rice, 14 parts of radish seed, the coptis
9 parts, 9 parts of gynostemma pentaphylla, 12 parts of sweet osmanthus, 14 parts of pseudo-ginseng, 12 parts of the red sage root, 15 parts of the Radix Astragali, 14 parts of fructus amomi, 6 parts of radix scutellariae, forrest silkvine stem or root 12
10 parts of part, 12 parts of japanese rose root, 9 parts of rhizoma zingiberis and Piper longum.The above-mentioned medicinal material of recipe quantity is taken to be broken into fine powder, saline cistanche polypeptide extract is removed
Outside, the ethanol solution for being 90% according to 7 times of addition volumetric concentrations of coarse powder gross weight, refluxing extraction three times, return time is 2h,
The medicinal extract for being concentrated to give that relative density is 1.20 at saline cistanche water extract, 60 DEG C is added in filtering, the filtrate of gained, that is, is obtained
Drug extract of the present invention.Clinically conventional Chinese medicine pharmaceutical dosage form can be prepared by preparation process thereof commonly used in the art, such as
Tablet or capsule.
The Chinese medicine composition and preparation technology of diarrhoea caused by comparative example one, a kind for the treatment of cancer chemotherapy：
Choose purposes of the CN103585546A Chinese medicine compositions in anti-diarrhea drug caused by treating cancer chemotherapy is prepared
Embodiment 6.
The pharmacodynamic study of Part III, saline cistanche polypeptide extract of the present invention
The inhibitory action of embodiment 6, medicine of the present invention to S180 tumor-bearing mice tumour growths
1st, modeling：It is experimental subjects to choose SPF grades of KM mouse of male, and body weight is 20-24g, and illumination/12 hour are black within 12 hours
Secretly, feed and water are freely absorbed.In addition to normal group, take and extract ascites under the S180 ascites mouse of inoculation 7 days, aseptic condition, use
Physiological saline adjusts cell concentration to 5 × 106/ml.S180 cell suspensions are only seeded in mouse right fore armpit with 0.2ml/
Subcutaneously.
2nd, animal packet and administration
After inoculation 24 hours, mouse is randomly divided into 5 groups respectively, every group 10, following medicines are given respectively：
Control group：Physiological saline
Saline cistanche polypeptide extract made from the embodiment of the present invention 3：20 mg/kg
The medicine group of the embodiment of the present invention 4：50 mg/kg
The medicine group of the embodiment of the present invention 5：50 mg/kg
Above-mentioned each medicine is intraperitoneal injection, 1 time a day, continuous 10 days.After last dose 24 hours, put to death small
Mouse, strips tumor mass and weighs, and calculates per cell mean, calculates tumour inhibiting rate, tumour inhibiting rate=(control group knurl weight-administration group knurl weight)/control
Group knurl weight × 100%.
Each pharmaceutical composition antineoplastic of table 1 is to Inhibition test in ascites tumor S180 bodies
Saline cistanche polypeptide extract produced by the present invention is to S180 tumor-bearing mice tumour growths it can be seen from upper table 1
With good inhibiting effect, with drug matching using the medicine group for constituting the present invention, it is to S180 tumor-bearing mice tumour growths
Inhibitory action become apparent, especially optimal with the medicine group of the embodiment of the present invention 5, inhibiting rate highest, illustrate the present invention medicine
Very strong effect is played in terms of tumor suppression.
Embodiment 7, medicine of the present invention and CN103585546A Chinese medicine compositions are to the Mice with Spleen diarrhea mould caused by rheum officinale
The therapeutic action of type
This experimental example be Mice with Spleen diarrhea model testing saline cistanche polypeptide extract of the present invention caused by rheum officinale and
The effect that medicine of the present invention is moved to mouse small intestine, the pharmacodynamic action under doses is observed, and is as a result sent out
Existing, the medicine can obviously improve its sign to the splenasthenic diarrhea mouse caused by rheum officinale, suppresses the gastrointestinal motility of mice with spleen deficiency, confrontation and lets out
Rush down.
Method：Male mice in kunming is taken, is grouped at random by body weight, every group 10, the following medicine of gavage：
Model group：Physiological saline
Saline cistanche polypeptide extract made from the embodiment of the present invention 3：20mg/kg (with 1g/ml decoction)
The medicine group of the embodiment of the present invention 4：50mg/kg (with 1g/ml decoction)
The medicine group of the embodiment of the present invention 5：50mg/kg (with 1g/ml decoction)
CN103585546A Chinese medicine composition groups：50mg/kg (with 1g/ml decoction)
In addition to Normal group, the equal gavage of remaining each group gives 10% rhubarb powder suspension, each 0.15mL/10g,
2 times a day, successive administration 15 days, causes Mice with Spleen model.From the 3rd day of self-supporting rhubarb powder suspension, medicine of the present invention
Group and CN103585546A Chinese medicine composition groups start gavage relative medicine, and one time a day, continuous 13 days.Model and normal control
Group is to isometric distilled water.In administration process, excrement and the sign change of mouse are observed at any time, is weighed before being administered daily small
The body weight of mouse.Mouse is administered once on the 16th day again in 12h fasting before experiment, and 1.5 hours charcoal ends to mouse stomach 5% are mixed after medicine
After suspension 0.1mL/10g body weight, 20min, mouse takes off cervical vertebra and put to death, and opens abdominal cavity and carefully separates mesenterium, clip upper end is certainly deep and remote
Door, intestinal tube is gently not added with traction and is laid on glass plate by the intestinal tube of lower end to ileocecus, with meter ruler measure total small intestinal length and
The distance that charcoal end is promoted.The last propulsion rate of charcoal is calculated by following equation.
Charcoal end propulsion rate (%)=charcoal end is in enteral actuating length/total small intestinal length × 100%
Therapeutic action of each pharmaceutical composition of table 2 to the Mice with Spleen diarrhea model caused by rheum officinale
Compared with Normal group▲▲P ＜ 0.01；Compared with model control group★P ＜ 0.05,★★P ＜ 0.01.
As a result：As shown in table 2,10% rhubarb powder suspension successive administration 16 days, model group and each administration group mouse
Start soft stool on the day of the modeling after 4h, some animals diarrhea, then occur within the 4th day anorexia, activity reduce, be slow in action,
The insufficiency of the spleen symptoms such as perpendicular hair hogback, hair color tarnishing.Normal group is then without above-mentioned phenomenon.In the 3rd day each administration group after modeling
The corresponding medicine of gavage, the hair color and the state of mind of each administration group mouse are better than model group.Rong's desert wherein produced by the present invention
The medicine group of Rong's polypeptide extract and the embodiment of the present invention is superior to CN103585546A Chinese medicine composition groups, with present invention implementation
The diarrhoea inhibiting rate highest of the medicine group of example 5, diarrhoea inhibition is most obvious.As a result medicine of the present invention is pointed out to be suspended rhubarb powder
Mice with spleen deficiency gastrointestinal motility caused by liquid accelerates have obvious antagonism, gastrointestinal motility function is tended to be normal.
Claims (8)
1. the method for saline cistanche polypeptide extract is prepared in a kind of powder from saline cistanche, it is characterised in that comprise the steps of：
(1) it is 9, the extraction time 50min at 50 DEG C to add NaOH solution to pH value toward saline cistanche powder, is then centrifuged for processing, i.e.,
Obtain saline cistanche albumen；
(2) the saline cistanche albumen obtained using step (1) configures the saline cistanche albumino reaction system of concentration of substrate 7%, adjusts meat
The pH value of desert cistanche albumino reaction system controls 65 DEG C of temperature to 6.5, adds enzymolysis protein enzyme of the enzyme bottom than 3.5%, constant temperature stirring
Enzymolysis, enzymolysis time is 3.5h, produces enzymolysis liquid, and described enzymolysis protein enzyme is papain；
(3) enzymolysis liquid obtained to step (2) carries out the 15min HCl solution for boiling destroy the enzyme treatment, 2mol/L being used after cooling
PH value is adjusted to neutrality, is then centrifuged for removing precipitation, takes supernatant, supernatant contains saline cistanche protein peptides；
(4) it is 45 DEG C in temperature, pH value is 8.0, pressure by the supernatant containing saline cistanche protein peptides obtained by step (3)
Under 0.8MPa, to select 500Da and l0000Da milipore filter to the meat desert of 500-10000Da molecular weight in saline cistanche protein peptides
Rong's polypeptide is separated, and obtains the medicinal extract for being concentrated to give that relative density is 1.20 at separating liquid, 60 DEG C, and as saline cistanche polypeptide is carried
Take thing.
2. saline cistanche polypeptide extract prepared by a kind of method described in use claim 1.
3. saline cistanche polypeptide extract as claimed in claim 2, it is characterised in that contain in described saline cistanche polypeptide extract
There is saline cistanche polypeptide 99%, the molecular weight of saline cistanche polypeptide is 500-10000Da.
4. saline cistanche polypeptide extract as claimed in claim 2 is for preparing the purposes in treatment sarcoma medicine.
5. saline cistanche polypeptide extract as claimed in claim 2 is for preparing the medicine suffered from diarrhoea caused by treating cancer chemotherapy
In purposes.
6. the purposes as described in claim 4 or 5, it is characterised in that the medicine is to be made up of the raw material of following parts by weight
Composition：5-16 parts of saline cistanche polypeptide extract, 7-9 parts of hawthorn, 5-8 parts of red yeast rice, 6-14 parts of radish seed, 3-9 parts of the coptis, strand
Blue 5-9 parts, 5-12 parts of sweet osmanthus, 5-14 parts of pseudo-ginseng, 8-12 parts of the red sage root, 9-15 parts of the Radix Astragali, 6-14 parts of fructus amomi, 4-6 parts of radix scutellariae, winged celestial being
6-10 parts of 6-12 parts of rattan, 5-12 parts of japanese rose root, 5-9 parts of rhizoma zingiberis and Piper longum.
7. purposes as claimed in claim 6, it is characterised in that the preparation method of the medicine includes following step：Take prescription
The above-mentioned medicinal material of amount is broken into except fine powder, saline cistanche polypeptide extract, and adding volumetric concentration according to 5-8 times of coarse powder gross weight is
75-90% ethanol solution, refluxing extraction three times, return time is 2-4h, filtering, and saline cistanche water extraction is added in the filtrate of gained
The medicinal extract for being concentrated to give that relative density is 1.20 at thing, 60 DEG C is taken, that is, obtains drug extract of the present invention.
8. purposes as claimed in claim 6, it is characterised in that described medicine is powder, aqua, tablet or capsule.
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| CN106265211B (en) * | 2015-06-03 | 2020-03-31 | 上海家化生物科技有限公司 | Cistanche salsa supernatant mixture, preparation method thereof and skin barrier repair effect thereof |
| KR101881142B1 (en) * | 2016-02-29 | 2018-07-23 | 한국 한의학 연구원 | Composition for preventing, improving or treating disease caused by side effect of anticancer agent comprising Cistanchis Herba extract as effective component |
| CN105648016A (en) * | 2016-03-30 | 2016-06-08 | 蔡庭守 | Herba cistanche small molecule peptide extract, preparation method and application thereof |
| CN106389541A (en) * | 2016-11-02 | 2017-02-15 | 杏辉天力（杭州）药业有限公司 | Cistanche deserticola extract and preparation method and application thereof |
| CN106860851A (en) * | 2017-02-09 | 2017-06-20 | 赵德润 | It is a kind of to prevent and treat rape pollen active peptide of prostatic disorders and preparation method thereof |
| CN107586821A (en) * | 2017-10-31 | 2018-01-16 | 亿利耐雀生物科技有限公司 | A kind of extracting method and purposes of saline cistanche polypeptide |
| CN109468358B (en) * | 2018-12-28 | 2021-05-28 | 河北肽丰生物科技有限公司 | Method for extracting cistanche chelating peptide |
| CN112047997B (en) * | 2020-08-10 | 2022-09-23 | 苏秀兰 | Cistanche polypeptide mixture, preparation method and application thereof |
| CN114600868A (en) * | 2022-02-22 | 2022-06-10 | 张帅军 | Cell preservation solution and application thereof in preservation of CIK cells |
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