CN104623623A - Preparation method of gecko maggot protein and application of gecko maggot protein to preparation of drug for treating uterine leiomyoma - Google Patents
Preparation method of gecko maggot protein and application of gecko maggot protein to preparation of drug for treating uterine leiomyoma Download PDFInfo
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Abstract
The invention discloses a preparation method of a gecko maggot protein and application of the gecko maggot protein to preparation of a drug for treating uterine leiomyoma. The preparation method comprises the following steps of puffing and smashing a gecko maggot in a normal-temperature puffing device; then, adding the smashed gecko maggot into an expanding solvent system extracting device to extract a crude gecko maggot protein; collecting a solution containing the crude gecko maggot protein, and filtering by using a micropore filter to remove impurities; carrying out ultrafiltration treatment by using an ultrafilter, and collecting a gecko maggot protein ultrafiltration solution; dissolving the ultrafiltration solution into acetonitrile, separating by using a solid-phase extraction column, and collecting an eluted component; dissolving the eluted component into a solvent system, and separating and purifying by using an ultrasonic stationary wave preparative liquid chromatography; and preparing freeze-dried protein powder by using a vacuum freeze-drying machine, wherein the freeze-dried protein powder can be used for preparing the drug for treating uterine leiomyoma. The purified gecko maggot protein separated by using the preparation method is a non-uniform protein, has the relative molecular weight of 10.9-12.6KPa and does not contain glycosyl; the pH value of an isoelectric point is 8.8-10.2; and the content of the purified gecko maggot protein is 96.7%.
Description
Technical field
The present invention relates to a kind of preparation method and novelty teabag thereof of medicine, especially the preparation method of Gekko Swinhonis maggot protein and the application in the anti-hysteromyoma medicine of preparation thereof.
Background technology
Hysteromyoma is also known as leiomyoma of uterus, and be the common a kind of benign tumors of female sex organs, major part is asymptomatic, and small number of patients shows as the symptoms such as vaginal hemorrhage, menoxenia, leucorrhoea grow in quantity.
Now conventional Therapeutic Method has: the excision of Drug therapy, palace laparoscopic surgery, tumor Excision etc., but the side effect that operative treatment brings is more.
CN102370851A discloses the Chinese herbal medicine for the treatment of hysteromyoma regulating menstruation, is made up of 12 taste Chinese herbal medicine such as Radix Salviae Miltiorrhizae, Radix Angelicae Sinensis, Flos Carthami, Flos Rosae Chinensis, Flos Celosiae Cristatae, Squama Manis, Flos Chrysanthemi, Herba Pteridis Multifidae, Spica Prunellaes; CN102319392A discloses a kind of Traditional Chinese Medicine Dropping Pill for the treatment of hysteromyoma and preparation method thereof, is made up of Chinese herbal medicine such as Concha Ostreae, Rhizoma Cyperi, Rhizoma Sparganii, Rhizoma Curcumae, Herba Hedyotidis Diffusaes; CN102327531A discloses a kind of Chinese patent medicine and Chinese medicine decoction of effective treatment hysteromyoma, is made up of 12 taste Chinese herbal medicine such as Radix Angelicae Sinensis, Semen Persicae, Rhizoma Curcumae, Rhizoma Cyperi, Semen Vaccariae, Rhizoma Sparganii; CN102247581A discloses a kind of medicine for the treatment of hysteromyoma, is pulverized form by 15 kinds of Chinese herbal medicine such as Rhizoma Cyperi, Radix Angelicae Sinensis, Rhizoma Chuanxiong, Semen Persicae, Rhizoma Sparganii, Radix Salviae Miltiorrhizae, Radix Scrophulariae, Cortex Moutan, Radix Glycyrrhizae; CN102302649A discloses a kind of Chinese medicine composition for the treatment of hysteromyoma, is made up of Rhizoma Dioscoreae Bulbiferae, Semen corchori, Caulis Cucurbitae, Exocarpium Citri Grandis, Radix Foeniculi, Nodus Nelumbinis Rhizomatis etc.
In existing Therapeutic Method, have and adopt GUIZHI FULING JIAONANG associating Mifepristone Treating Uterine Leimyoma to achieve certain curative effect, also have fragrant rib ball in employing " Ji the side of seating " to add taste and treat hysteromyoma; Also the doctor had achieves certain curative effect from intending four conjunctions gram tumor side's treatment hysteromyoma; The useful mifepristone of Drug therapy, danazol, tamoxifen androgen and promoting sexual gland hormone.
201210315707.2 techniques disclosing artificial culture Gekko Swinhonis maggot, to the invention describes with Gekko Swinhonis etc. as raw material, grow, cultivate through training, prepared by post processing become Gekko Swinhonis maggot.Process reconfiguration Gekko Swinhonis gene of the present invention, the Gekko Swinhonis maggot composition of generation has the function of anti-hysteromyoma, can produce in batches.
In principle, above-mentioned several Chinese medicine preparation may be used to the preparation for the treatment of hysteromyoma medicine.But, because aforesaid several prescription Chinese crude drug is various, often kind of Chinese herbal medicine due to the difference of plantation region, weather, kind, collecting time, the process of preparing Chinese medicine and processing method, therefore, the concordance ensureing product is difficult to, the also quality of restive product in suitability for industrialized production.
Summary of the invention
The object of this invention is to provide a kind of preparation method of Gekko Swinhonis maggot protein and the application in the anti-hysteromyoma medicine of preparation thereof, utilize Gekko Swinhonis maggot for raw material, through separation and purification protein, again with the medicine of the various dosage form of this protein preparation, this process is briefly clear, drug ingredient is clear and definite, and concordance is good, and curative effect is obvious.
The object of the invention is to be achieved through the following technical solutions:
(1) by the expanded pulverizing under normal temperature condition in room temperature expanding apparatus (200520108339.X) of 1000g Gekko Swinhonis maggot.
In this step, described bulking pressure is 0.1 ~ 5MPa, and the time is 10 ~ 60min.
(2) material after expanded pulverizing joins in expanded solvents system extraction element (201210179076.6), adds expanded solvents and is forced into 2 ~ 10MPa, forms expanded solvents system, extracts Gekko Swinhonis maggot protein crude product.
In this step, described expanded solvents is by 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL methanol and 100 ~ 300gNH
3h
2o forms or is made up of 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL ethanol and 100 ~ 300g ammonium chloride.
In this step, described Extracting temperature is 10 ~ 20 DEG C, and the time is 10 ~ 30min.
(3) collect the solution containing Gekko Swinhonis maggot protein crude product and cross with microfilter and filter to remove impurity.
In this step, the aperture of described microfilter is 0.5 ~ 50 μm.
(4) use ultrafilter hyperfiltration treatment, collect the Gekko Swinhonis maggot protein ultrafiltration solution containing corresponding molecular weight.
In this step, the aperture of described ultrafilter is 50 ~ 100nm.
(5) ultrafiltrate is dissolved in suitable solvent and is separated with solid-phase extraction column, collects eluting composition.
In this step, described solvent is acetonitrile.
(6) by eluting component dissolves in suitable dicyandiamide solution, with ultrasonic standing wave preparative liquid chromatogram (200920274485.8) separation and purification.
In this step, described dicyandiamide solution is made up of n-butyl alcohol-ethyl acetate-water (volume ratio 1.25:3.75:5) or 4.5% Polyethylene Glycol-7% Dextran T 500 (weight ratio) forms.
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder.
In this step, described vacuum lyophilization temperature is-30 ~ 50 DEG C, the time is 18 ~ 26h.
(8) give different medicinal auxiliary material, be prepared into injection, oral agents, suppository respectively, can be used for preparing anti-hysteromyoma medicine.
The purification Gekko Swinhonis maggot protein be separated in the technical program records non-homogeneous protein in polyacrylamide gel electrophoresis, two-dimensional polyacrylamide gel electrophoresis and agar polysaccharide immunoelectrophoresis spectrum; Recording relative molecular weight by SDS-polyacrylamide gel electrophoresis and sephadex G-75 gel filtration is 10.9 ~ 12.6KPa; Sending out its isoelectric point, IP of mensuration with isoelectrofocusing is pH8.8 ~ 10.2; Conventional chemistry analysis result shows it not containing glycosyl; The Gekko Swinhonis maggot protein content of determined by ultraviolet spectrophotometry purification is 96.7%.
Detailed description of the invention
Below in conjunction with embodiment, technical scheme of the present invention is further described; but do not limit to so; everyly technical solution of the present invention modified or equivalent to replace, and not departing from the spirit and scope of technical solution of the present invention, all should be encompassed in protection scope of the present invention.
embodiment 1:
(1) 1000g Gekko Swinhonis maggot dry product is loaded in room temperature expanding apparatus (200520108339.X), to being filled with carbon dioxide in expanding apparatus and being forced into 0.1 ~ 5MPa, time 10 ~ 60min, then release pressure is to normal pressure rapidly, and Gekko Swinhonis maggot is by expanded one-tenth powder body;
(2) the Gekko Swinhonis maggot powder body after expanded pulverizing joins in expanded solvents system extraction element (201210179076.6), adds 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL methanol and 100 ~ 300gNH
3h
2o, is forced into 2 ~ 10MPa, forms expanded solvents system, and keep temperature 10 ~ 20 DEG C, time 10 ~ 30min, the proteolytic in Gekko Swinhonis maggot powder body, is then depressurized to normal pressure, generates Gekko Swinhonis maggot protein crude product and methanol-NH
3h
2the mixed liquor of O;
(3) collect the solution containing Gekko Swinhonis maggot protein crude product, filter place to go solid impurity with the microfilter in 0.5 ~ 50 μm, aperture;
(4) the solution ultrafilter hyperfiltration treatment of aperture 50 ~ 100nm, pressure 0.3 ~ 0.8MPa, temperature is room temperature, collects the Gekko Swinhonis maggot protein ultrafiltration solution containing corresponding molecular weight;
(5) ultrafiltration solution 20% acetonitrile dissolves, C
18solid-phase extraction column 50 ~ 100% acetonitrile activating pretreatments, add 0.02 ~ 0.1% trifluoroacetic acid-water balance, the ultrafiltration solution upper prop dissolved with 10% acetonitrile solution, wash with the acetonitrile gradient of 10 ~ 80% is de-under room temperature, flow velocity 1 ~ 10mL/min, collects the eluting composition of associated gradients;
(6) by eluting component dissolves in 4.5% Polyethylene Glycol-7% Dextran T 500 (weight ratio) dicyandiamide solution, with ultrasonic standing wave preparative liquid chromatogram (200920274485.8) separation and purification, ultrasonic power 0.2 ~ 3.0KW, frequency 500 ~ 1000KHz, flow velocity 50 ~ 500mL/min, UV-detector wavelength 260 ~ 280nm;
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder in temperature-30 ~ 50 DEG C, time 18 ~ 26h drying.
embodiment 2:
(1) 1000g Gekko Swinhonis maggot dry product is loaded in room temperature expanding apparatus (200520108339.X), to being filled with carbon dioxide in expanding apparatus and being forced into 0.1 ~ 5MPa, time 10 ~ 60min, then release pressure is to normal pressure rapidly, and Gekko Swinhonis maggot is by expanded one-tenth powder body;
(2) the Gekko Swinhonis maggot powder body after expanded pulverizing joins in expanded solvents system extraction element (201210179076.6), adds 5Kg liquid carbon dioxide, 300mL methanol and 200gNH
3h
2o, is forced into 5MPa, forms expanded solvents system, and keep temperature 15 DEG C, time 20min, the proteolytic in Gekko Swinhonis maggot powder body, is then depressurized to normal pressure, generates Gekko Swinhonis maggot protein crude product and methanol-NH
3h
2the mixed liquor of O;
(3) collect the solution containing Gekko Swinhonis maggot protein crude product, filter place to go solid impurity with the microfilter in 30 μm, aperture;
(4) the solution ultrafilter hyperfiltration treatment of aperture 100nm, pressure 0.5MPa, temperature is room temperature, collects the Gekko Swinhonis maggot protein ultrafiltration solution containing corresponding molecular weight;
(5) ultrafiltration solution 20% acetonitrile dissolves, C
18solid-phase extraction column 80% acetonitrile activating pretreatment, adds 0.05% trifluoroacetic acid-water balance, the ultrafiltration solution upper prop dissolved with 10% acetonitrile solution, and wash with the acetonitrile gradient of 10 ~ 80% is de-under room temperature, flow velocity 5mL/min, collects the eluting composition of associated gradients;
(6) by eluting component dissolves in 4.5% Polyethylene Glycol-7% Dextran T 500 (weight ratio) dicyandiamide solution, with ultrasonic standing wave preparative liquid chromatogram (200920274485.8) separation and purification, ultrasonic power 1.0KW, frequency 500KHz, flow velocity 200mL/min, UV-detector wavelength 260nm;
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder in temperature-30 ~ 40 DEG C, time 20h drying.
embodiment 3:
(1) 1000g Gekko Swinhonis maggot dry product is loaded in room temperature expanding apparatus (200520108339.X), to being filled with carbon dioxide in expanding apparatus and being forced into 0.1 ~ 5MPa, time 10 ~ 60min, then release pressure is to normal pressure rapidly, and Gekko Swinhonis maggot is by expanded one-tenth powder body;
(2) the Gekko Swinhonis maggot powder body after expanded pulverizing joins in expanded solvents system extraction element (201210179076.6), add 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL ethanol and 100 ~ 300g ammonium chloride, be forced into 2 ~ 10MPa, form expanded solvents system, keep temperature 10 ~ 20 DEG C, time 10 ~ 30min, the proteolytic in Gekko Swinhonis maggot powder body, then be depressurized to normal pressure, generate the mixed liquor of Gekko Swinhonis maggot protein crude product and ethanol-ammonium chloride;
(3) collect the solution containing Gekko Swinhonis maggot protein crude product, filter place to go solid impurity with the microfilter in 0.5 ~ 50 μm, aperture;
(4) the solution ultrafilter hyperfiltration treatment of aperture 50 ~ 100nm, pressure 0.3 ~ 0.8MPa, temperature is room temperature, collects the Gekko Swinhonis maggot protein ultrafiltration solution containing corresponding molecular weight;
(5) ultrafiltration solution 20% acetonitrile dissolves, C
18solid-phase extraction column 50 ~ 100% acetonitrile activating pretreatments, add 0.02 ~ 0.1% trifluoroacetic acid-water balance, the ultrafiltration solution upper prop dissolved with 10% acetonitrile solution, wash with the acetonitrile gradient of 10 ~ 80% is de-under room temperature, flow velocity 1 ~ 10mL/min, collects the eluting composition of associated gradients;
(6) by eluting component dissolves in n-butyl alcohol-ethyl acetate-water (volume ratio 1.25:3.75:5) dicyandiamide solution, with ultrasonic standing wave preparative liquid chromatogram (200920274485.8) separation and purification, ultrasonic power 0.2 ~ 3.0KW, frequency 500 ~ 1000KHz, flow velocity 50 ~ 500mL/min, UV-detector wavelength 260 ~ 280nm;
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder in temperature-30 ~ 50 DEG C, time 18 ~ 26h drying.
embodiment 4:
(1) 1000g Gekko Swinhonis maggot dry product is loaded in room temperature expanding apparatus (200520108339.X), to being filled with carbon dioxide in expanding apparatus and being forced into 2MPa, time 40min, then release pressure is to normal pressure rapidly, and Gekko Swinhonis maggot is by expanded one-tenth powder body;
(2) the Gekko Swinhonis maggot powder body after expanded pulverizing joins in expanded solvents system extraction element (201210179076.6), add 6Kg liquid carbon dioxide, 200mL ethanol and 150g ammonium chloride, be forced into 6MPa, form expanded solvents system, keep temperature 20 DEG C, time 10min, the proteolytic in Gekko Swinhonis maggot powder body, then be depressurized to normal pressure, generate the mixed liquor of Gekko Swinhonis maggot protein crude product and ethanol-ammonium chloride;
(3) collect the solution containing Gekko Swinhonis maggot protein crude product, filter place to go solid impurity with the microfilter in 50 μm, aperture;
(4) the solution ultrafilter hyperfiltration treatment of aperture 100nm, pressure 0.5MPa, temperature is room temperature, collects the Gekko Swinhonis maggot protein ultrafiltration solution containing corresponding molecular weight;
(5) ultrafiltration solution 20% acetonitrile dissolves, C
18solid-phase extraction column 60% acetonitrile activating pretreatment, adds 0.08% trifluoroacetic acid-water balance, the ultrafiltration solution upper prop dissolved with 10% acetonitrile solution, and wash with the acetonitrile gradient of 10 ~ 80% is de-under room temperature, flow velocity 8mL/min, collects the eluting composition of associated gradients;
(6) by eluting component dissolves in n-butyl alcohol-ethyl acetate-water (volume ratio 1.25:3.75:5) dicyandiamide solution, with ultrasonic standing wave preparative liquid chromatogram (200920274485.8) separation and purification, ultrasonic power 2.0KW, frequency 1000KHz, flow velocity 300mL/min, UV-detector wavelength 280nm;
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder in temperature-30 ~ 50 DEG C, time 18h drying.
embodiment 5:
Get Gekko Swinhonis maggot lyophilized protein powder 100 ~ 150g, lecithin 1 ~ 15g, liquid paraffin 200 ~ 350g, three is stirred, pasty state is ground well into 30 ~ 60r/min with grinder, separately get oleum sapii 100 ~ 200g heat in water-bath 40 ~ 60 DEG C fusing after, be cooled to 25 ~ 38 DEG C, pour in the Gekko Swinhonis maggot protein-lecithin-liquid paraffin mixture ground well and stir rapidly, be cast in while hot in suppository moulds, condense to 10 ~ 20 DEG C, from suppository moulds, take out the Gekko Swinhonis maggot protein suppository being molding, arrange smooth rear packed products.
embodiment 6:
Get Gekko Swinhonis maggot lyophilized protein powder 120g, lecithin 10g, liquid paraffin 250g, three is stirred, pasty state is ground well into 60r/min with grinder, separately get oleum sapii 150g heat in water-bath 60 DEG C fusing after, be cooled to 30 DEG C, pour in the Gekko Swinhonis maggot protein-lecithin-liquid paraffin mixture ground well and stir rapidly, be cast in while hot in suppository moulds, condense to 15 DEG C, from suppository moulds, take out the Gekko Swinhonis maggot protein suppository being molding, arrange smooth rear packed products.
embodiment 7:
Prepared by lipidosome injection:
(1) after the 1mol Phosphatidylserine glass planar method of forming makes thin film, add 1000 ~ 3000L normal saline hydration-treated, then use ultrasound wave at frequency 500 ~ 2000KHz, power 0.01 ~ 5KW process 1 ~ 5min.
(2) 0.1 ~ 1000mLCaCl that above-mentioned solution 1 ~ 100mL joins content 0.001 ~ 0.01mol/L is got
2in solution, under rotating speed 30 ~ 75r/min condition, stir 10 ~ 30min, then under 25 ~ 50 DEG C of conditions, leave standstill 30 ~ 120min.
(3) product is placed in centrifuge, at rotating speed 1500 ~ 3000r/min centrifugalize 5 ~ 30min, after being taken out from centrifuge by centrifugal thing, add 2 ~ 50g Gekko Swinhonis maggot protein mix homogeneously, under 10 ~ 25 DEG C of conditions, leave standstill 10 ~ 120min.
(4) above-mentioned object adds in the normal saline solution 0.1 ~ 10L of the ethylenediaminetetraacetic acid containing 0.01 ~ 10mol/L, and mix homogeneously leaves standstill 30 ~ 180min, obtains unilamellar liposome.
(5) get 1 ~ 10mg unilamellar liposome to mix with 10 ~ 200mL normal saline solution, preparation becomes injection.
embodiment 8:
Prepared by lipidosome injection:
(1), after the 1mol Phosphatidylserine glass planar method of forming makes thin film, 2000L normal saline hydration-treated is added, then with ultrasound wave at frequency 1000KHz, power 2KW process 5min.
(2) 1000mLCaCl that above-mentioned solution 50mL joins content 0.005mol/L is got
2in solution, under rotating speed 45r/min condition, stir 30min, then under 30 DEG C of conditions, leave standstill 60min.
(3) product is placed in centrifuge, at rotating speed 2000r/min centrifugalize 20min, after being taken out from centrifuge by centrifugal thing, add 20g Gekko Swinhonis maggot protein mix homogeneously, under 15 DEG C of conditions, leave standstill 60min.
(4) above-mentioned object adds in the normal saline solution 5L of the ethylenediaminetetraacetic acid containing 1mol/L, and mix homogeneously leaves standstill 80min, obtains unilamellar liposome.
(5) get 5mg unilamellar liposome to mix with 100mL normal saline solution, preparation becomes injection.
embodiment 9:
The preparation of enteric coated capsule:
Get 10 ~ 100g Gekko Swinhonis maggot protein, 500 ~ 2000g pregelatinized starch and 5-50g lactose fully to mix, being packed into preparation in hydroxypropyl emthylcellulose enteric coated capsule becomes Gekko Swinhonis maggot protein enteric coated capsule.
embodiment 10:
The preparation of enteric coated capsule:
Get 50g Gekko Swinhonis maggot protein, 1000g pregelatinized starch and 30g lactose fully to mix, being packed into preparation in hydroxypropyl emthylcellulose enteric coated capsule becomes Gekko Swinhonis maggot protein enteric coated capsule.
embodiment 11:
Hysteromyoma diagnostic criteria:
(1) change in menstrual cycle characteristics, some patients can touch hypogastric region enclosed mass, the symptoms such as companion's leucorrhoea grow in quantity, urine urgency-frequency, dysuria, infertile, secondary anemia.
(2) B ultrasonic prompting: uterus increases, matter is hard.
(3) detect uterine cavity to increase or distortion.
(4) examine when scraping and touch crowning in uterine cavity.
(5) B ultrasonic and uteroscopy can assist diagnosis.
Therapeutic Method:
Patient needs strict restriction wine, by body weight 60Kg, every day liposome local injection 2 doses, every agent 250 ~ 300 μ g, 2 weeks 1 courses for the treatment of, or every day vagina administration suppository 1 piece, 2 weeks 1 courses for the treatment of, or take enteric coated capsule every day 2 times, each 2,2 weeks 1 courses for the treatment of.
Criterion of therapeutical effect:
Recovery from illness: symptom all disappears, ovary chocolate cyst disappears substantially through the local such as gynecologial examination, ultrasound diagnosis pelvic lump sign;
Effective: symptom disappears substantially, ovary chocolate cyst reduces more than 1/2 through gynecologial examination, ultrasound diagnosis Endometrial adenocarcinoma three footpath sum, clinical symptoms local or all disappear;
Effective: symptom alleviates, ovary chocolate cyst reduces more than 1/3 through gynecologial examination, ultrasound diagnosis cyst three footpath sum, clinical symptoms partial disappearance or alleviate;
Invalid: more than 1 course for the treatment of of taking medicine continuously, ultrasound diagnosis cyst three footpath sum is without change or reduce less than 1cm, and subjective symptoms alleviates or without change.
Therapeutic outcome:
Claims (10)
1. a preparation method for Gekko Swinhonis maggot protein, is characterized in that described preparation method step is as follows:
(1) by the expanded pulverizing under normal temperature condition in room temperature expanding apparatus of 1000g Gekko Swinhonis maggot;
(2) material after expanded pulverizing joins in expanded solvents system extraction element, adds by 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL methanol and 100 ~ 300gNH
3h
2the expanded solvents that O forms or is made up of 1 ~ 10Kg liquid carbon dioxide, 100 ~ 500mL ethanol and 100 ~ 300g ammonium chloride is also forced into 2 ~ 10MPa, forms expanded solvents system, extracts Gekko Swinhonis maggot protein crude product;
(3) collect the solution containing Gekko Swinhonis maggot protein crude product and cross with microfilter and filter to remove impurity;
(4) use ultrafilter hyperfiltration treatment, collect Gekko Swinhonis maggot protein ultrafiltration solution;
(5) ultrafiltrate is dissolved in acetonitrile and is separated with solid-phase extraction column, collects eluting composition;
(6) by eluting component dissolves by n-butyl alcohol-ethyl acetate-water dicyandiamide solution that 1.25:3.75:5 forms or is made up of by weight 4.5:7 Polyethylene Glycol-Dextran T 500 by volume, use ultrasonic standing wave preparative liquid chromatogram separation and purification;
(7) the Gekko Swinhonis maggot protein vacuum freeze drier after purification is prepared into lyophilized protein powder.
2. the preparation method of Gekko Swinhonis maggot protein according to claim 1, it is characterized in that described bulking pressure is 0.1 ~ 5MPa, the time is 10 ~ 60min.
3. the preparation method of Gekko Swinhonis maggot protein according to claim 1, it is characterized in that described Extracting temperature is 10 ~ 20 DEG C, the time is 10 ~ 30min.
4. the preparation method of Gekko Swinhonis maggot protein according to claim 1, is characterized in that the aperture of described microfilter is 0.5 ~ 50 μm.
5. the preparation method of Gekko Swinhonis maggot protein according to claim 1, is characterized in that the aperture of described ultrafilter is 50 ~ 100nm.
6. the preparation method of Gekko Swinhonis maggot protein according to claim 1, is characterized in that described vacuum lyophilization temperature is-30 ~ 50 DEG C, the time is 18 ~ 26h.
7. the Gekko Swinhonis maggot protein that prepared by the arbitrary described method of claim 1-6 is preparing the application in anti-hysteromyoma medicine.
8. the application of Gekko Swinhonis maggot protein according to claim 7 in the anti-hysteromyoma medicine of preparation, it is characterized in that the dosage form of described anti-hysteromyoma medicine is suppository, preparation method is as follows:
Get Gekko Swinhonis maggot lyophilized protein powder 100 ~ 150g, lecithin 1 ~ 15g, liquid paraffin 200 ~ 350g, three is stirred, pasty state is ground well into 30 ~ 60r/min with grinder, separately get oleum sapii 100 ~ 200g heat in water-bath 40 ~ 60 DEG C fusing after, be cooled to 25 ~ 38 DEG C, pour in the Gekko Swinhonis maggot protein-lecithin-liquid paraffin mixture ground well and stir rapidly, be cast in while hot in suppository moulds, condense to 10 ~ 20 DEG C, from suppository moulds, take out the Gekko Swinhonis maggot protein suppository being molding.
9. the application of Gekko Swinhonis maggot protein according to claim 7 in the anti-hysteromyoma medicine of preparation, it is characterized in that the dosage form of described anti-hysteromyoma medicine is lipidosome injection, preparation method is as follows:
(1) after the 1mol Phosphatidylserine glass planar method of forming makes thin film, add 1000 ~ 3000L normal saline hydration-treated, then use ultrasound wave at frequency 500 ~ 2000KHz, power 0.01 ~ 5KW process 1 ~ 5min;
(2) 0.1 ~ 1000mLCaCl that above-mentioned solution 1 ~ 100mL joins content 0.001 ~ 0.01mol/L is got
2in solution, under rotating speed 30 ~ 75r/min condition, stir 10 ~ 30min, then under 25 ~ 50 DEG C of conditions, leave standstill 30 ~ 120min;
(3) product is placed in centrifuge, at rotating speed 1500 ~ 3000r/min centrifugalize 5 ~ 30min, after being taken out from centrifuge by centrifugal thing, add 2 ~ 50g Gekko Swinhonis maggot protein mix homogeneously, under 10 ~ 25 DEG C of conditions, leave standstill 10 ~ 120min;
(4) above-mentioned object adds in the normal saline solution 0.1 ~ 10L of the ethylenediaminetetraacetic acid containing 0.01 ~ 10mol/L, and mix homogeneously leaves standstill 30 ~ 180min, obtains unilamellar liposome;
(5) get 1 ~ 10mg unilamellar liposome to mix with 10 ~ 200mL normal saline solution, preparation becomes injection.
10. the application of Gekko Swinhonis maggot protein according to claim 7 in the anti-hysteromyoma medicine of preparation, it is characterized in that the dosage form of described anti-hysteromyoma medicine is enteric coated capsule, preparation method is as follows:
Get 10 ~ 100g Gekko Swinhonis maggot protein, 500 ~ 2000g pregelatinized starch and 5-50g lactose fully to mix, be packed in hydroxypropyl emthylcellulose enteric coated capsule, preparation becomes Gekko Swinhonis maggot protein enteric coated capsule.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102702308A (en) * | 2012-06-02 | 2012-10-03 | 哈尔滨工业大学 | Expansion solvent system extraction device and process for separating toad maggot proteins by using same |
| CN102783461A (en) * | 2012-08-31 | 2012-11-21 | 哈尔滨工业大学 | Process for artificially breeding wall gecko maggots |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102702308A (en) * | 2012-06-02 | 2012-10-03 | 哈尔滨工业大学 | Expansion solvent system extraction device and process for separating toad maggot proteins by using same |
| CN102783461A (en) * | 2012-08-31 | 2012-11-21 | 哈尔滨工业大学 | Process for artificially breeding wall gecko maggots |
Non-Patent Citations (2)
| Title |
|---|
| 胭苒: "子宫肌瘤，我该如何处置你", 《食品与健康》 * |
| 陈国才等: "云南SLS-壁虎组织注射液研究简报", 《中药通报》 * |
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Application publication date: 20150520 |