CN102657847A - Application of toad maggot protein in medicine used for treating rectal cancer - Google Patents
Application of toad maggot protein in medicine used for treating rectal cancer Download PDFInfo
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- CN102657847A CN102657847A CN2012101791190A CN201210179119A CN102657847A CN 102657847 A CN102657847 A CN 102657847A CN 2012101791190 A CN2012101791190 A CN 2012101791190A CN 201210179119 A CN201210179119 A CN 201210179119A CN 102657847 A CN102657847 A CN 102657847A
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Abstract
The invention discloses application of toad maggot protein in a medicine used for treating rectal cancer and belongs to the technical field of medicines. Aiming at the problems of the existing medicine used for treating rectal cancer, the invention combines the toad maggot protein and appropriate pharmaceutical necessities for use and provides a novel application of the toad maggot protein in a medicine used for treating rectal cancer. The invention is designed to aim at the special symptom of rectal cancer; the content of toad maggot protein which is extracted and purified is 96.5-98.1%; and the toad maggot protein with the purity is used for preparing a medicine in various forms. The medicine has definite ingredients, good consistency, controllable quality, good curative effect for rectal cancer and convenience for use.
Description
Technical field
The invention belongs to medical technical field, relate to the application of a kind of Bufo siccus maggot protein in treatment rectal cancer medicine.
Background technology
Rectal cancer is to be cancerated and formed by the rectal tissue cell, and its morbidity and social environment, dietary habit, inherited genetic factors are relevant, are one of digestive tract common malignancy.
CN1850183 (200610041831.9) discloses a kind of chemotherapeutics of treating rectal cancer, is made up of 5-fluorouracil, liquid paraffin, vaseline, lanoline, span, tween, herb liquid.
ZL200610104558.X discloses a kind of Chinese medicine composition of treating rectal cancer, is made up of 16 kinds of Chinese herbal medicine such as Radix Ginseng, Radix Angelicae Sinensis, Ganoderma, Colophonium, Rhizoma Chuanxiong, Radix Sophorae Flavescentis, Calculus Bovis, Squama Maniss.
CN101584824 (200810100694.0) discloses a kind of Chinese medicine of treating rectal cancer, is made up of 16 kinds of Chinese herbal medicine such as Fructus Akebiae, the Radix Aucklandiae, Caulis Sargentodoxae, Herba Hedyotidis Diffusae, Radix Sophorae Flavescentis, Radix Salviae Miltiorrhizae, Fructus Mume, Herba Pteridis Multifidae, Herba Scutellariae Barbataes.
ZL201010140829.3 discloses a kind of Chinese medicine composition of treating rectal cancer and preparation method thereof, is made up of 25 kinds of Chinese herbal medicine such as Poria, Radix Sophorae Tonkinensis, Radix Trichosanthis, Radix Scutellariae, Radix Sophorae Flavescentis, Radix Arnebiae (Radix Lithospermi), Flos Lonicerae, Herba Lobeliae Chinensis, Rhizoma Curcumae, Rhizoma Corydalis.
CN1568681 discloses and has adopted Chinese medicine to cultivate the method for Bufo siccus maggot, in batches suitability for industrialized production Bufo siccus maggot.
Aforesaid in principle several kinds of Chinese medicine preparation may be used to treat the preparation of rectal cancer medicine; But; Because aforesaid several prescription Chinese medicines are various; Every kind of Chinese herbal medicine is because the difference of plantation region, weather, kind, collecting time, the process of preparing Chinese medicine and processing method is difficult to guarantee consistency of product and quality control in suitability for industrialized production.
Summary of the invention
To the problem that existing treatment rectal cancer medicine exists, the present invention is used in combination Bufo siccus maggot protein with suitable pharmaceutical necessities, and the new purposes of a kind of Bufo siccus maggot protein in treatment rectal cancer medicine is provided.
Treatment rectal cancer medicine of the present invention prepares according to following step:
One, extract Bufo siccus maggot protein:
(1) the dry Bufo siccus maggot of 1000g is put into the room temperature expanding apparatus, charge into 1～10kg nitrogen, 0.1～0.5kg ammonia, be forced into 0.2～1.0Mpa, time 10～30min, gas in the abrupt release device then, the Bufo siccus maggot is by expanded pulverizing;
(2) expanded comminuting matter joins in the expanded solvents system extraction element, charges into 200～2000g carbon dioxide, 100～500g ethanol and 100～200g dimethyl formamide, is forced into 1～8Mpa; 10～25 ℃ of temperature; Time 10～60min, release pressure is collected mixed liquor to normal pressure then;
(3) mixed liquor separates with 1500～5000r/min with seperator, collects liquid phase;
(4) collect liquid phase and filter, get filtrate with membrane aperture 0.5～50 μ m Microfilter;
(5) filtrate is injected in the solvent resistant column of saphacryls-100HR under pH=3～12 conditions, filters;
(6) filtrating is dissolved in the dicyandiamide solution of being made up of according to the volume ratio of 4:3:3:2 chloroform-ethanol-methanol-water; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram; Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow velocity 50～500mL/min, UV-detector wavelength 280nm, separation of pure dissolves Bufo siccus maggot protein;
(7) purification Bufo siccus maggot protein under vacuum 200～800Kpa, temperature-50～30 ℃, the condition of time 6～24h, is dried to the freeze-dried protein finished product that contains moisture 5～8% with vacuum freeze drier;
Two, mix: mix according to proportioning, be processed into the treatment rectal cancer medicine of various dosage forms.
Bufo siccus maggot protein is the biological activity protein of extraction separation purification from the Bufo siccus maggot; Molecular structure is clear, and medicinal potency ratio is higher, high specificity; Toxic reaction relatively a little less than; Be difficult in vivo producing accumulating, have the awareness of molecule with the affinity of body inner recipient, and almost do not have alienation biological metabolism toxicity.
The present invention is directed to the rectal cancer characteristic symptom and design; A kind of Bufo siccus maggot protein content that extracts purification reaches 96.5～98.1%; The Bufo siccus maggot protein of this purity of reuse is prepared into the medicine of various dosage forms, and this drug ingredient is clear and definite, high conformity; Quality controllable, to rectal cancer disease good effect and easy to use.
Description of drawings
Fig. 1 is the structural representation of expanded solvents system extraction element.
The specific embodiment
The specific embodiment one: this embodiment is treated the medicine of rectal cancer according to following method preparation:
(1) the dry Bufo siccus maggot of 1000g is put into room temperature expanding apparatus (ZL200520108339.X); Charge into 1～10kg nitrogen, 0.1～0.5kg ammonia, be forced into 0.2～1.0Mpa, time 10～30min; Gas in the abrupt release device then, the Bufo siccus maggot is by expanded pulverizing;
(2) expanded comminuting matter joins in the expanded solvents system extraction element, charges into 200～2000g carbon dioxide, 100～500g ethanol and 100～200g dimethyl formamide, is forced into 1～8Mpa; 10～25 ℃ of temperature; Time 10～60min, release pressure is collected mixed liquor to normal pressure then;
(3) mixed liquor separates with 1500～5000r/min with seperator, collects liquid phase;
(4) collect liquid phase and filter, get filtrate with membrane aperture 0.5～50 μ m Microfilter;
(5) filtrate is injected in the solvent resistant column of saphacryls-100HR under pH=3～12 conditions, filters;
(6) filtrating is dissolved in the dicyandiamide solution of being made up of according to the volume ratio of 4:3:3:2 chloroform-ethanol-methanol-water; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram (ZL200920274485.8); Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow velocity 50～500mL/min, UV-detector wavelength 280nm, separation of pure dissolves Bufo siccus maggot protein;
(7) purification Bufo siccus maggot protein under vacuum 200～800Kpa, temperature-50～30 ℃, the condition of time 6～24h, is dried to the freeze-dried protein finished product that contains moisture 5～8% with vacuum freeze drier;
(8) get above-mentioned freeze-dried protein 10～15g, lecithin 0.1～1.5g, liquid paraffin 20～50g, stir, grind well into pasty state; Other gets oleum sapii 100～200g and in water-bath, is heated to 45～55 ℃ of fusing postcooling to 35～38 ℃; Pour in the protein that grinds well into pasty state and lecithin and the liquid paraffin mixture and grind rapidly evenly, be cast in while hot in the suppository mould, condense to 10～20 ℃; From the suppository mould, take out molding Bufo siccus maggot protein suppository, packed products.
As shown in Figure 1; The system of expanded solvents described in this embodiment extraction element is made up of spherical expanded solvents system extractor 1, head tank 2, solid-liquid separator 3, gas-solid separator 4, finished product jar 5, external source carbon dioxide cylinder 6, carbon dioxide storage tank 7, carbon dioxide circulating pressure pump 8 and solvent tank 9; Spherical expanded solvents system extractor 1 is connected with head tank 2, solid-liquid separator 3, carbon dioxide circulating pressure pump 8 and solvent tank 9 respectively through pipeline; Solid-liquid separator 3 is connected with gas-solid separator 4; Gas-solid separator 4 is connected with finished product jar 5 and carbon dioxide storage tank 7 respectively through pipeline, and carbon dioxide storage tank 7 is connected with external source carbon dioxide cylinder 6 and carbon dioxide circulating pressure pump 8 respectively through pipeline.
The specific embodiment two: this embodiment is treated the medicine of rectal cancer according to following method preparation:
(1) with the bright Bufo siccus maggot of 1000g with beater at 500～1000r/min, the 10～30min that pulls an oar, become slurry;
(2) adding contains 0.01～0.5% aqueous sodium carbonate, 5～20L in the slurry, stirs, and places 30～120min at normal temperatures;
(3), collect liquid phase with centrifuge centrifugal 10～30min under 1500～5000r/min condition;
(4) filter above-mentioned liquid phase with membrane aperture 0.2～5.0 μ m Microfilter, collect filtrate;
(5) filtrating is injected in the saphacryls-100HR solvent resistant column to cross in pH=3～12 filters filtrate;
(6) filtrate is dissolved in the dicyandiamide solution of being made up of according to the volume ratio of 4::3:3:2 chloroform-ethanol-methanol-water; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram; Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow 50～500mL/min; UV-detector wavelength 280nm, continuous purification separate Bufo siccus maggot protein;
(7) at vacuum 50～500Kpa, temperature-30～30 ℃ is dried under time 6～24h condition and contains moisture 5～8% finished products purification Bufo siccus maggot protein with vacuum freeze drier;
(8) join after 100g lecithin and 30～80g cholesterol are mixed and becomes I liquid in 500～1000ml ether, 10～50g Bufo siccus maggot protein joins in 3～10mmol/L PBS stirring and dissolving and processes II liquid, and I liquid is mixed with II liquid afterwards with ultrasound wave at power 0.1～0.5KW; Frequency 300～500KHz, sonic oscillation 10～30min under the room temperature heats 50～80 ℃ then in rotary evaporator; Vacuum 100～500Kpa, rotating speed 30～120r/min are evaporated to and present gel, take off the evaporation flask be placed on the gyrate shaker make the gel phase inversion with 30～200r/min vibration after; To evaporate flask again puts into water-bath and heats 60～80 ℃; Remove clean ether, again material is put into the inherent 15000～50000r/min ultracentrifugation of supercentrifuge 30～60min, separate and remove the Bufo siccus maggot protein that does not coat; Precipitate is with 1～2L deionized water wash 2～3 times; Centrifuge is sloughed water at 150～750r/min, promptly gets Bufo siccus maggot protein liposome, and collecting packing promptly gets.
The rectal cancer staging:
0 phase: cancer is confined to mucous layer, no lymphatic metastasis;
The I phase: tumor is confined in the muscularis propria, no lymphatic metastasis;
The II phase: neoplasm invasiveness surpasses muscularis propria, but does not have lymphatic metastasis;
The III phase: lymphatic metastasis;
The IV phase: metastasis (liver, lung etc.) or peritoneum shift.
Therapeutic Method:
The patient needs strict restriction to drink.By body weight 70kg, supp anal administration every day, 3 of every days, each 1, or the liposome local injection, 2 doses of every days, every dose 250～300 μ g, a course of treatment 2 Mondays.
The therapeutic outcome grade scale:
One, short term effect (4 weekends are relatively preceding with treatment)
1, alleviate fully: the focus complete obiteration, no new focus occurs;
2, part is alleviated: 50% before the treatment that focus is dwindled or littler, and no new focus occurs, and the neither one focus increased when many focal diseases became;
3, stable phase: 1. slightly alleviate: focus is dwindled area 20% or is increased area 25%, and subjective symptoms is improved, and physical basal conditions rises; 2. basicly stable: focus is dwindled less than 25% or is increased area and be no more than more than 25%, and subjective symptoms is improved, and physical basal conditions rises;
4, expansion: increase more than 25% before single area or the treatment of a plurality of focus total area ratio, or new pathological changes occurs, clinical symptoms increases the weight of, and physical basal conditions descends.
Two, intermediate period treatment: (5 week～1 year)
1, alleviates no tumor existence fully;
2, part is alleviated: 1. corpus carcinosus is returned to area before the treatment more than 50% or new focus occurs; 2. corpus carcinosus does not have increase or dwindles doing well,improving or disappearance; 3. physical basal conditions rises;
3, transfer and expansion: the cause of disease kitchen range enlarges development, new MET occurs, and clinical symptoms increases the weight of, and physical situation descends.
Three, late result: (1 year～5 years)
1, no tumor existence;
2, band tumor existence;
3, death.
Therapeutic outcome: see table 1
Table 1
| Short term effect | Alleviate fully | Part is alleviated | The stable phase expansion | Obvious effective rate % | Effective percentage % |
| Patient 22 people | 15 | 5 | 1 1 | 95.5 | 91 |
| Mid-term curative effect | Alleviate fully | Part is alleviated | Shift and expansion | Obvious effective rate % | Effective percentage % |
| Patient 19 people | 11 | 5 | 3 | 84 | 84 |
| Late result | No tumor existence | The existence of band tumor | Dead | Obvious effective rate % | Effective percentage % |
| Patient 20 |
7 | 8 | 5 | 75 | 75 |
Above embodiment has been merely the present invention has been further described, and scope of the present invention does not receive the limitation of the embodiment of lifting.
Claims (9)
1. the application of Bufo siccus maggot protein in treatment rectal cancer medicine.
2. the application of Bufo siccus maggot protein according to claim 1 in treatment rectal cancer medicine is characterized in that said treatment rectal cancer medicine is the suppository of being processed by Bufo siccus maggot protein 10～15g, lecithin 0.1～1.5g, liquid paraffin 20～50g, oleum sapii 100～200g.
3. the application of Bufo siccus maggot protein according to claim 1 and 2 in treatment rectal cancer medicine is characterized in that said Bufo siccus maggot protein obtains according to following method:
(1) the dry Bufo siccus maggot of 1000g is put into the room temperature expanding apparatus, charge into 1～10kg nitrogen, 0.1～0.5kg ammonia, be forced into 0.2～1.0Mpa, time 10～30min, gas in the abrupt release device then, the Bufo siccus maggot is by expanded pulverizing;
(2) expanded comminuting matter joins in the expanded solvents system extraction element, charges into 200～2000g carbon dioxide, 100～500g ethanol and 100～200g dimethyl formamide, is forced into 1～8Mpa; 10～25 ℃ of temperature; Time 10～60min, release pressure is collected mixed liquor to normal pressure then;
(3) mixed liquor separates with 1500～5000r/min with seperator, collects liquid phase;
(4) collect liquid phase and filter, get filtrate with membrane aperture 0.5～50 μ m Microfilter;
(5) filtrate is injected in the solvent resistant column of saphacryls-100HR under pH=3～12 conditions, filters;
(6) filtrating is dissolved in the dicyandiamide solution; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram; Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow velocity 50～500mL/min, UV-detector wavelength 280nm, separation of pure dissolves Bufo siccus maggot protein;
(7) purification Bufo siccus maggot protein under vacuum 200～800Kpa, temperature-50～30 ℃, the condition of time 6～24h, is dried to the freeze-dried protein finished product that contains moisture 5～8% with vacuum freeze drier.
4. the application of Bufo siccus maggot protein according to claim 3 in treatment rectal cancer medicine is characterized in that said dicyandiamide solution is made up of according to the volume ratio of 4:3:3:2 chloroform-ethanol-methanol-water.
5. the application of Bufo siccus maggot protein according to claim 1 in treatment rectal cancer medicine is characterized in that said treatment rectal cancer medicine is the liposome of being processed by 100g lecithin, 30～80g cholesterol, 10～50g Bufo siccus maggot protein.
6. according to claim 1 or the application of 5 described Bufo siccus maggot protein in treatment rectal cancer medicine, it is characterized in that said Bufo siccus maggot protein obtains according to following method:
(1) with the bright Bufo siccus maggot of 1000g with beater at 500～1000r/min, the 10～30min that pulls an oar, become slurry;
(2) adding mass concentration in the slurry is 0.01～0.5% aqueous sodium carbonate, 5～20L, stirs, and places 30～120min at normal temperatures;
(3), collect liquid phase with centrifuge centrifugal 10～30min under 1500～5000r/min condition;
(4) filter above-mentioned liquid phase with membrane aperture 0.2～5.0 μ m Microfilter, collect filtrate;
(5) filtrating is injected in the saphacryls-100HR solvent resistant column to cross in pH=3～12 filters filtrate;
(6) filtrate is dissolved in the dicyandiamide solution, pumps in the ultrasonic standing wave liquid phantom preparing chromatogram ultrasonic standing wave power 0.2～5.0KW; Frequency 500～1000kHz; Flow 50～500mL/min, UV-detector wavelength 280nm, continuous purification separate Bufo siccus maggot protein;
(7) at vacuum 50～500Kpa, temperature-30～30 ℃ is dried under time 6～24h condition and contains moisture 5～8% finished products purification Bufo siccus maggot protein with vacuum freeze drier.
7. the application of Bufo siccus maggot protein according to claim 6 in treatment rectal cancer medicine is characterized in that said dicyandiamide solution is made up of according to the volume ratio of 4::3:3:2 chloroform-ethanol-methanol-water.
8. the application of Bufo siccus maggot protein according to claim 1 in treatment rectal cancer medicine is characterized in that said treatment rectal cancer medicine prepares according to following method:
One, extract Bufo siccus maggot protein:
(1) the dry Bufo siccus maggot of 1000g is put into the room temperature expanding apparatus, charge into 1～10kg nitrogen, 0.1～0.5kg ammonia, be forced into 0.2～1.0Mpa, time 10～30min, gas in the abrupt release device then, the Bufo siccus maggot is by expanded pulverizing;
(2) expanded comminuting matter joins in the expanded solvents system extraction element, charges into 200～2000g carbon dioxide, 100～500g ethanol and 100～200g dimethyl formamide, is forced into 1～8Mpa; 10～25 ℃ of temperature; Time 10～60min, release pressure is collected mixed liquor to normal pressure then;
(3) mixed liquor separates with 1500～5000r/min with seperator, collects liquid phase;
(4) collect liquid phase and filter, get filtrate with membrane aperture 0.5～50 μ m Microfilter;
(5) filtrate is injected in the solvent resistant column of saphacryls-100HR under pH=3～12 conditions, filters;
(6) filtrating is dissolved in the dicyandiamide solution of being made up of according to the volume ratio of 4:3:3:2 chloroform-ethanol-methanol-water; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram; Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow velocity 50～500mL/min, UV-detector wavelength 280nm, separation of pure dissolves Bufo siccus maggot protein;
(7) purification Bufo siccus maggot protein under vacuum 200～800Kpa, temperature-50～30 ℃, the condition of time 6～24h, is dried to the freeze-dried protein finished product that contains moisture 5～8% with vacuum freeze drier;
Two, mix: mix according to the said proportioning of claim 1, be processed into the treatment rectal cancer medicine of various dosage forms.
9. the application of Bufo siccus maggot protein according to claim 1 in treatment rectal cancer medicine is characterized in that said Bufo siccus maggot method for extracting protein is:
(1) with the bright Bufo siccus maggot of 1000g with beater at 500～1000r/min, the 10～30min that pulls an oar, become slurry;
(2) adding mass concentration in the slurry is 0.01～0.5% aqueous sodium carbonate, 5～20L, stirs, and places 30～120min at normal temperatures;
(3), collect liquid phase with centrifuge centrifugal 10～30min under 1500～5000r/min condition;
(4) filter above-mentioned liquid phase with membrane aperture 0.2～5.0 μ m Microfilter, collect filtrate;
(5) filtrating is injected in the saphacryls-100HR solvent resistant column to cross in pH=3～12 filters filtrate;
(6) filtrate is dissolved in the dicyandiamide solution of being made up of according to the volume ratio of 4::3:3:2 chloroform-ethanol-methanol-water; Pump in the ultrasonic standing wave liquid phantom preparing chromatogram; Ultrasonic standing wave power 0.2～5.0KW, frequency 500～1000kHz, flow 50～500mL/min; UV-detector wavelength 280nm, continuous purification separate Bufo siccus maggot protein;
(7) at vacuum 50～500Kpa, temperature-30～30 ℃ is dried under time 6～24h condition and contains moisture 5～8% finished products purification Bufo siccus maggot protein with vacuum freeze drier.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1093584A (en) * | 1993-04-12 | 1994-10-19 | 钟俊生 | Hepatocarcinoma medicinal liquid and preparation method thereof |
| CN1568681A (en) * | 2004-05-10 | 2005-01-26 | 孔繁伟 | Method for cultivating toad evildoers using traditional Chinese medicine |
| CN102091318A (en) * | 2011-01-14 | 2011-06-15 | 哈尔滨工业大学 | Toad peptide antibiotics separated from toad maggots and preparation method of antibacterial drugs thereof |
-
2012
- 2012-06-02 CN CN201210179119.0A patent/CN102657847B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1093584A (en) * | 1993-04-12 | 1994-10-19 | 钟俊生 | Hepatocarcinoma medicinal liquid and preparation method thereof |
| CN1568681A (en) * | 2004-05-10 | 2005-01-26 | 孔繁伟 | Method for cultivating toad evildoers using traditional Chinese medicine |
| CN102091318A (en) * | 2011-01-14 | 2011-06-15 | 哈尔滨工业大学 | Toad peptide antibiotics separated from toad maggots and preparation method of antibacterial drugs thereof |
Non-Patent Citations (1)
| Title |
|---|
| 赵冬梅和刘素宾: "动物药在***中的应用及其药理研究", 《中医药学刊》, vol. 22, no. 4, 30 April 2004 (2004-04-30), pages 757 - 759 * |
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