CN102973605B - Toad skin extract dry powder inhalant and preparation method and applications thereof - Google Patents
Toad skin extract dry powder inhalant and preparation method and applications thereof Download PDFInfo
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Abstract
The invention relates to a dry powder inhalant, and in particular relates to a dry powder inhalant made of a toad skin extract, a preparation method of the dry powder inhalant and applications thereof in preparing a medicament for treating lung tumor. The toad skin extract dry powder inhalant is characterized by consisting of a toad skin extract and a medically acceptable adjuvant, wherein the mass ratio of the toad skin extract to the adjuvant is 1:0 to 1:110. The dry powder inhalant is prepared with the toad skin extract, and the inhalant can directly acts on a lung infected part, thus reducing the occurrence of toxic and side effects, improving the bioavailability of the medicament, not only overcoming the shortcomings that an oral preparation is slow in action, has liver first pass effect and systemic toxic and side effects, is relatively low in preparation bioavailability, and the like, and at the same time solving the problems that an injection is inconvenient to use, poor in patient compliance, and the like.
Description
technical field:
The present invention relates to Foradil Aerolizer formoterol fumarate, specifically Cutis Bufonis extract make Foradil Aerolizer formoterol fumarate, the preparation method of described Foradil Aerolizer formoterol fumarate and the application in preparation treatment lung tumors medicine.
background technology:
Cinobufacin is the extract that the dry skin of Bufonidae (Bufonidae) animal Bufo siccus Bufobufo gargarizans Cantor obtains through extraction purification, there is effect of removing toxic substances, detumescence, pain relieving, in being used for the treatment of, the disease such as late tumor, chronic hepatitis B, its biological active substances is bufotoxin class (bufotoxins) and hydrolyzate bufotalin class (bufageins), bufotenine class (bufoteinines) etc. mainly.Current cinobufacin injection, oral liquid, tablet have been applied to clinical, have good curative effect to hepatocarcinoma, pulmonary carcinoma, gastric cancer etc.Especially HUACHANSU ZHUSHEYE, from the water soluble preparation of Skin of Bufo bufo gargarizans through being processed into, treatment primary hepatocarcinoma and medium and advanced lung cancer obtain the comprehensive effective percentage of 44% and 56% respectively, and tumor body minification is respectively 10% and 16%, and has synergism to chemotherapy and radiation.Although the clinical untoward reaction of HUACHANSU ZHUSHEYE is less, but still there is intravenous drip and there will be local vein reaction, cause instillation vein to shrink spasm and cause pain, long-time stimulus causes vein nonspecific inflammation; Dermoreaction, main manifestations is urticaria and the infringement of skin blister sample; Secondly, there is the cardiovascular system reactions such as premature beat, sinus bradycardia, sinus tachycardia, atrioventricular block in small part patient, the possible cause of generation is the cardiotoxicity that bufotalin a small amount of in cinobufacin produces.In addition, injection needs to use in hospital, and patient compliance is poor.And cinobufacin oral formulations need through the first pass effect of liver, onset is slow, and preparation bioavailability is lower.
Lung has great surface area, and alveolar wall is made up of cell monolayer, makes drug molecule very easily enter blood, and therefore lung inhalation has the plurality of advantages such as rapid-action, untoward reaction is little.Foradil Aerolizer formoterol fumarate (drypowder inhalers in lung inhalation, DPIs) propellant and solvent is not used, the air-flow produced by patient's autonomous respiration makes drug powder be atomized, and then make drug powder be delivered to pulmonary, have application convenient, absorb rapidly, cross without liver head, feature that rapid release and targeting are located, be a kind of novel drug-supplying system.
The sickness rate of pulmonary carcinoma rises rapidly in worldwide, has become first of the cancer cause of the death.Current antitumor drug great majority are all drug administration by injection, and this Formulations for systemic administration mode is unfavorable for that drug accumulation is in tumor locus, and cause the curative effect of medicine to reduce, untoward reaction increases.
Adopt site-specific delivery of drugs antitumor drug to be transported to the tumor patient part of human body, medicine is improved in the concentration of patient part, to the toxic and side effects of reduction medicine, the therapeutic effect of raising medicine, there is great clinical value.Adopt focus Cutis Bufonis extract directly can being transmitted pulmonary carcinoma through the inhalant of lung administering mode, be conducive to improving Cutis Bufonis extract to the curative effect of pulmonary carcinoma and reduce the toxic and side effects of its normal tissue.The research at present with the toad skin extract dry powder inhalant of resisting tumor of lung effect has no report.
summary of the invention:
Goal of the invention: pulmonary's dry powder inhaler formulations that an object of the present invention is to provide Cutis Bufonis extract and preparation method thereof.Another object of the present invention is to provide the application of Foradil Aerolizer formoterol fumarate of the present invention in preparation treatment lung tumors medicine.
For foregoing invention object, the invention provides following technical scheme:
A kind of toad skin extract dry powder inhalant, is characterized in that it is made up of Cutis Bufonis extract and medically acceptable adjuvant, and Cutis Bufonis extract (according to gained solids quality after drying) is 1:0-1:110 with the mass ratio of described adjuvant.
Described a kind of toad skin extract dry powder inhalant, be add by Cutis Bufonis extract the powder that adjuvant makes, powder agglomeration forms re-dispersable particles; Or Cutis Bufonis extract load forms granule on granules of accessories surface; The particle diameter of above-mentioned two kinds of granules is with D
50meter is between 0.5-1000 micron.
Described Cutis Bufonis extract, the extract that the skin for Bufonidae animal Bufo siccus or Bufo melanostictus etc. obtains through extraction purification.The preparation method of Cutis Bufonis extract is:
Get Cutis Bufonis 1 part, add 2-100 times of water gaging, backflow or decoct or ultrasonic or microwave or lixiviate or seepage pressure effects 1-10 time, each 0.1-48 hour, merge Aqueous extracts, being concentrated into relative density is 1.01-1.50(60-100 DEG C), add ethanol (65%-100%, W/W) precipitation, ethanol content is made to reach 50%(W/W) more than, standing or centrifugal or filtration treatment, gets supernatant and reclaims ethanol, obtains Cutis Bufonis extract after obtaining Cutis Bufonis extract or concentrated, drying.
Or, get Cutis Bufonis 1 part, add 2-100 times of water gaging, backflow or decoct or ultrasonic or microwave or lixiviate or seepage pressure effects 1-10 time, each 0.1-48 hour, merge Aqueous extracts, being concentrated into relative density is 1.01-1.50(60-100 DEG C), add ethanol (65%-100%, W/W) precipitate, ethanol content is made to reach 50%(W/W) more than, standing or centrifugal or filtration treatment, get supernatant and reclaim ethanol extremely without alcohol taste, add ethanol (65%-100%, W/W) precipitate, ethanol content is made to reach 60%(W/W) more than, standing or centrifugal or filtration treatment, get supernatant and reclaim ethanol extremely without alcohol taste, obtain Cutis Bufonis extract or concentrate, Cutis Bufonis extract is obtained after drying.
Or, get Cutis Bufonis 1 part, add 2-100 times of water gaging, backflow or decoct or ultrasonic or microwave or lixiviate or seepage pressure effects 1-10 time, each 0.1-48 hour, merge Aqueous extracts, through the ultrafilter membrane ultrafiltration of molecular weight 500-5000, after obtaining Cutis Bufonis extract or concentrated, drying, obtain Cutis Bufonis extract.
Or, get Cutis Bufonis 1 part, add the certain density ethanol (1%-100%, W/W) of 2-100 times amount, backflow or ultrasonic or microwave or lixiviate or seepage pressure effects 1-10 time, each 0.1-48 hour, merge extractive liquid, reclaims ethanol to without alcohol taste, adds 0-100 times of water, standing or centrifugal or filtration treatment, obtains Cutis Bufonis extract after obtaining Cutis Bufonis extract or concentrated, drying.
Or, get Cutis Bufonis 1 part, add the certain density ethanol (1%-100%, W/W) of 2-100 times amount, backflow or ultrasonic or microwave or lixiviate or seepage pressure effects 1-10 time, each 0.1-48 hour, merge extractive liquid, reclaims ethanol to without alcohol taste, adds 0-100 times of water, through the ultrafilter membrane ultrafiltration of molecular weight 500-5000, after obtaining Cutis Bufonis extract or concentrated, drying, obtain Cutis Bufonis extract.
One or more mixture in sugar, alcohol, aminoacid, phospholipid, surfactant, cyclodextrin, polymer substance, magnesium stearate, stearic acid, hard ester fumaric acid sodium, micropowder silica gel, Pulvis Talci selected by described adjuvant.
Described sugar, can select one or more mixture in galactose, lactose, glucose, fructose, sucrose, trehalose, Raffinose; Described alcohol, can select one or more mixture in mannitol, xylitol, maltose alcohol, sorbitol; Described aminoacid, can select one or more in glycine, Aspartic Acid, alanine, tryptophan, isoleucine, threonine, glutamic acid, phenylalanine, leucine, cystine, lysine, proline, arginine; Described phospholipid, can select one or more in soybean phospholipid, lecithin, phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE, synthetic phospholipid; Described surfactant, can select pulmonary surfactant as one or more mixture in dipalmitoyl phosphatidyl choline, DLPC, cholesterol; Described polymer substance, the macromolecule meeting safety requirements can be selected, one or more mixture in preferred biodegradable polymer substance such as albumin, starch, polylactic acid, poly lactic-co-glycolic acid, Polyethylene Glycol, poloxamer, hyaluronic acid, hyaluronate sodium, sodium alginate.
Preferred mode to be described adjuvant be suction lactose granule mean diameter is at 45-220 micron and the Micronised lactose mean diameter mixture at 1-10 micron, and wherein the mass ratio of lactose granule and Micronised lactose is 1000:1-1:1.Most preferred mode is the mass ratio of lactose granule and Micronised lactose is 100:1-7:1.
Prepare a method for toad skin extract dry powder inhalant, its preparation process is as follows:
By Cutis Bufonis extract by pulverize or spraying dry makes powder, be packaged in capsule or bubble-cap or Diskus; By Cutis Bufonis extract by pulverize or spraying dry makes powder, powder is mixed homogeneously with adjuvant and is made suction powder, is packaged in capsule or bubble-cap or Diskus; Or Cutis Bufonis extract is mixed homogeneously with adjuvant, by pulverize or spraying dry makes suction powder, is packaged in capsule or bubble-cap or Diskus; Or by partial supplementary material mix homogeneously in Cutis Bufonis extract and prescription, by pulverize or spraying dry makes powder, powder with remain adjuvant in prescription and mix homogeneously and make suction powder, be packaged in capsule or bubble-cap or Diskus.
Prepare a method for toad skin extract dry powder inhalant, its preparation process is as follows:
By Cutis Bufonis extract by pulverize or spraying dry makes powder, powder is mixed homogeneously with adjuvant, and formation re-dispersable particles of reuniting, is packaged in capsule or bubble-cap or Diskus; Or Cutis Bufonis extract is mixed homogeneously with adjuvant, by pulverize or spraying dry makes powder, reunites and form re-dispersable particles, be packaged in capsule or bubble-cap or Diskus; Or by partial supplementary material mix homogeneously in Cutis Bufonis extract and prescription, by pulverize or spraying dry makes powder, powder with remain adjuvant in prescription and mix homogeneously, formation re-dispersable particles of reuniting, be packaged in capsule or bubble-cap or Diskus; Or by Cutis Bufonis extract by pulverize or spraying dry makes powder, powder load forms granule on granules of accessories surface, is packaged in capsule or bubble-cap or Diskus.
A kind of toad skin extract dry powder inhalant is preparing the application in lung tumors medicine.
beneficial effect:
1, lung has great surface area, and alveolar wall is made up of cell monolayer, makes drug molecule very easily enter blood, and therefore lung inhalation has the plurality of advantages such as rapid-action, untoward reaction is little.Foradil Aerolizer formoterol fumarate (drypowder inhalers in lung inhalation, DPIs) propellant and solvent is not used, the air-flow produced by patient's autonomous respiration makes drug powder be atomized, and then make drug powder be delivered to pulmonary, have application convenient, absorb rapidly, cross without liver head, feature that rapid release and targeting are located, be a kind of novel drug-supplying system.There is the infection being used for treating respiratory tract and pulmonary about Foradil Aerolizer formoterol fumarate at present, but have no the treatment at lung tumors.Meanwhile, the research with the toad skin extract dry powder inhalant of resisting tumor of lung effect also has no report.Current Cutis Bufonis extract injection, oral liquid, tablet have been applied to clinical, have good effect to hepatocarcinoma, pulmonary carcinoma, gastric cancer etc.Especially HUACHANSU ZHUSHEYE, from the water soluble preparation of Skin of Bufo bufo gargarizans through being processed into, treatment primary hepatocarcinoma and medium and advanced lung cancer obtain the comprehensive effective percentage of 44% and 56% respectively, and tumor body minification is respectively 10% and 16%, and has synergism to chemotherapy and radiation.Although the clinical untoward reaction of HUACHANSU ZHUSHEYE is less, but still there is intravenous drip and there will be local vein reaction, cause instillation vein to shrink spasm and cause pain, long-time stimulus causes vein nonspecific inflammation; Dermoreaction, main manifestations is urticaria and the infringement of skin blister sample; Secondly, there is the cardiovascular system reactions such as premature beat, sinus bradycardia, sinus tachycardia, atrioventricular block in small part patient, the possible cause of generation is the cardiotoxicity that bufotalin a small amount of in cinobufacin produces.In addition injection needs to use in hospital, and patient compliance is poor.And Cutis Bufonis extract oral formulations need through the first pass effect of liver, onset is slow, and preparation bioavailability is lower.Use the Foradil Aerolizer formoterol fumarate of Cutis Bufonis extract mainly for lung tumors, directly act on affected part, there is not the first pass effect of liver simultaneously, rapid-action, and there is no above-mentioned untoward reaction, to new drug development, there is great prospect.
2, because Cutis Bufonis extract is mixture, wherein containing indole alkaloids, cinobufagin, bufogenin etc., find in practical operation, the specific adjuvant adding certain proportioning can overcome the reunion that the electrostatic interaction, cohesiveness etc. that produce when extract is crushed to certain particle diameter cause, and reduces hygroscopicity.Applicant is known through great many of experiments, and particularly adjuvant is selected the mixture of lactose granule and Micronised lactose or adopted powder agglomeration technology can make the excellent Foradil Aerolizer formoterol fumarate of intake performance.
Detailed description of the invention
Below in conjunction with example, the present invention is described in further detail, but scope of the present invention is not by any restriction of these examples.
Embodiment 1
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 10 times of water gagings, decoct 2 times, each 1.5 hours, merging filtrate, being concentrated into relative density is 1.1-1.2 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 60%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leave standstill 48 hours, filter, reclaim ethanol to thick paste, 60 DEG C of dryings, obtain Cutis Bufonis extract.
2, the preparation of inhalant: adopt comminution by gas stream to D Cutis Bufonis extract
50below 5 microns, with particle diameter 45-70 micron lactose in mass ratio 1:10 mix, drug powder is packaged in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 2
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 12 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2-1.3 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 70%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, spraying dry, obtains Cutis Bufonis extract powder, powder D
50below 5 microns.
2, the preparation of inhalant: Cutis Bufonis extract powder, particle diameter are mixed in the ratio of the leucine 10:100:1 in mass ratio of 20-50 micron at the lactose of 45-80 micron, particle diameter, is packaged in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 3
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 30 times of water gagings, microwave radiation exaraction 3 times, each 1 hour, merge extractive liquid, through the ultrafilter membrane ultrafiltration of molecular weight 1000, ultrafiltrate spraying dry obtains Cutis Bufonis extract powder, powder D
50below 5 microns.
2, the preparation of inhalant: Cutis Bufonis extract powder packets is incapsulated or in bubble-cap or Diskus, obtains inhalant.
Embodiment 4
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis clean, add 12 times amount 60%(W/W) ethanol, reflux, extract, 2 times, each 2 hours, merge extractive liquid, reclaims ethanol extremely without alcohol taste, deepfreeze 48 hours, filter, add the active carbon (W/V) of 0.3%, boil 30 minutes, filter, spraying dry, obtains Cutis Bufonis extract powder.
2, the preparation of inhalant: the ratio of Cutis Bufonis extract powder, lactose, mannitol, glycine, DLPC, lecithin, magnesium stearate 1:5:5:2:0.5:0.5:0.1 in mass ratio mixed, comminution by gas stream is to powder D
50below 5 microns, drug powder is packaged in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 5
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, add the dehydrated alcohol of 20 times amount, reflux, extract, 2 times, each 1 hour, merge extractive liquid, reclaimed ethanol, through the ultrafilter membrane ultrafiltration of molecular weight 3000, ultrafiltrate vacuum concentration to proportion is 1.20 (45 DEG C), and extractum 50 DEG C of vacuum dryings, obtain Cutis Bufonis extract.
2, the preparation of inhalant: Cutis Bufonis extract, lactose, cholesterol are mixed in the ratio of 1:8:1, comminution by gas stream is to powder D
50below 5 microns, drug powder is packaged in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 6
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, adds the 60%(W/W of 15 times amount) ethanol, reflux, extract, 3 times, each 1 hour, merge extractive liquid, reclaim ethanol extremely without alcohol taste, concentrated solution adds 2 times amount water precipitating, leave standstill 24 hours, high speed centrifugation, gets supernatant and obtains Cutis Bufonis extract.
2, the preparation of inhalant: respectively solid quality 5 times contained in above-mentioned Cutis Bufonis extract, lactose, polyethylene glycol 6000, the albumin of 1 times, 2 times are added in Cutis Bufonis extract, spraying dry, powder D
50below 5 microns, drug powder is packaged in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 7
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, adds the 10%(W/W of 10 times amount) ethanol, reflux, extract, 2 times, each 1 hour, merge extractive liquid, reclaim ethanol extremely without alcohol taste, concentrated solution high speed centrifugation, gets supernatant and obtains Cutis Bufonis extract, spraying dry, obtains Cutis Bufonis extract, powder D
50below 5 microns.
2, the preparation of inhalant: Cutis Bufonis extract drug powder, particle diameter are mixed in the ratio of the mannitol of 10-90 micron, micronized silica gel 10:89:1 in mass ratio, is packaged into after mix homogeneously in capsule or bubble-cap or Diskus, obtains inhalant.
Embodiment 8
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, adds the 50%(W/W of 10 times amount) ethanol, reflux, extract, 2 times, each 1 hour, merge extractive liquid, reclaims ethanol extremely without alcohol taste, concentrated solution high speed centrifugation, get supernatant and obtain Cutis Bufonis extract, spraying dry, obtain Cutis Bufonis extract powder.
2, the preparation of inhalant: the ratio of Cutis Bufonis extract drug powder, mannitol, leucine 10:100:3 in mass ratio mixed, add water after mixing appropriate, spraying dry obtains D
50being the powder of 4.8 microns, with there being the rotary blender of polymer inner liner to mix, making loose aggregate, the D of aggregate
50be 390 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 9
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, adds the 40%(W/W of 10 times amount) ethanol, reflux, extract, 3 times, each 0.5 hour, merge extractive liquid, reclaims ethanol extremely without alcohol taste, concentrated solution high speed centrifugation, get supernatant and obtain Cutis Bufonis extract, spraying dry, powder D
50below 5 microns.
2, the preparation of inhalant: the ratio of Cutis Bufonis extract powder, mannitol, leucine 10:80:2 in mass ratio being mixed, with there being the rotary blender of polymer inner liner to mix, making loose aggregate, the D of aggregate
50be 120 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 10
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 20 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2-1.3 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 70%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, spraying dry, powder D
50below 5 microns.
2, the preparation of inhalant: by Cutis Bufonis extract powder, German U.S. agent happy InhaLac 70 lactose, Micronised lactose (D
50=5.1 microns) in mass ratio the ratio of 5:90:5 mix, be packaged in capsule or bubble-cap or Diskus after mix homogeneously, obtain inhalant.
3, inhalant pulmonary deposition ratio is investigated
Investigate according to People's Republic of China's version States Pharmacopoeia specifications method in 2010, with bufothionine content meter in preparation, its pulmonary deposition ratio reaches 32.6%, and place after 18 months under room temperature condition, in preparation, the pulmonary deposition ratio of bufothionine does not have significant change.
Embodiment 11
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 20 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2-1.3 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 70%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, spraying dry, powder D
50below 5 microns.
2, the preparation of inhalant: by Cutis Bufonis extract powder, German U.S. agent happy InhaLac 70 lactose, Micronised lactose (D
50=4.9 microns) in mass ratio the ratio of 3:95:1 mix, be packaged in capsule or bubble-cap or Diskus after mix homogeneously, obtain inhalant.
3, inhalant pulmonary deposition ratio is investigated
Investigate according to People's Republic of China's version States Pharmacopoeia specifications method in 2010, with bufothionine content meter in preparation, its pulmonary deposition ratio reaches 30.2%, and place after 18 months under room temperature condition, in preparation, the pulmonary deposition ratio of bufothionine does not have significant change.
Embodiment 12
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 20 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2-1.3 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 70%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, spraying dry, powder D
50below 5 microns.
2, the preparation of inhalant: by Cutis Bufonis extract powder, German U.S. agent happy InhaLac 70 lactose, Micronised lactose (D
50=5.5 microns) in mass ratio the ratio of 3:77:11 mix, be packaged in capsule or bubble-cap or Diskus after mix homogeneously, obtain inhalant.
3, inhalant pulmonary deposition ratio is investigated
Investigate according to People's Republic of China's version States Pharmacopoeia specifications method in 2010, with bufothionine content meter in preparation, its pulmonary deposition ratio reaches 29.8%, and place after 18 months under room temperature condition, in preparation, the pulmonary deposition ratio of bufothionine does not have significant change.
Embodiment 13
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 10 times of water gagings, decoct 2 times, each 1 hour, merging filtrate, being concentrated into relative density is 1.3-1.4 (85 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 60%, leave standstill 48 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 80%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, spraying dry, powder D
50below 5 microns.
2, the preparation of inhalant: Cutis Bufonis extract powder, leucine are mixed in the ratio of 1:5, the D of granule after mixing
50be 292 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 14
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 6 times of water gagings, decoct 5 times, each 1 hour, merging filtrate, being concentrated into relative density is 1.05 (70 DEG C), be cooled to 40 DEG C, add alcohol settling, make alcohol content reach 65%, leave standstill 48 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 70%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, concentrating under reduced pressure, vacuum drying obtains Cutis Bufonis extract.
2, the preparation of inhalant: Cutis Bufonis extract is mixed with the ratio of leucine in 1:1, comminution by gas stream, powder D
50below 5 microns, this powder is mixed with the mannitol of 9 times amount again, the D of granule after mixing
50be 328 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 15
1, the preparation of Cutis Bufonis extract: get dry maxima skin and clean, add 30 times of water gagings, decoct 1 hour, it is 1.30 (60 DEG C) that extracting solution is concentrated into relative density, is cooled to 20 DEG C, adds alcohol settling, make alcohol content reach 70%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 90%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, concentrating under reduced pressure, vacuum drying obtains Cutis Bufonis extract.
2, the preparation of inhalant: the ratio of Cutis Bufonis extract and poloxamer 10:1 is in mass ratio mixed, comminution by gas stream, powder D
50below 5 microns, this powder is mixed with the lactose granule of 8 times amount, the magnesium stearate of 1 times amount again, the D of granule after mixing
50be 653 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 16
1, the preparation of Cutis Bufonis extract: get Cutis Bufonis and clean, add 10 times of water gagings, decoct 1 hour, it is 1.20 (50 DEG C) that extracting solution is concentrated into relative density, is cooled to room temperature, adds alcohol settling, make alcohol content reach 50%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 70%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, concentrating under reduced pressure, vacuum drying obtains Cutis Bufonis extract.
2, the preparation of inhalant: by Cutis Bufonis extract comminution by gas stream, powder D
50below 5 microns, this powder is mixed with the lactose granule of 20 times amount again, the D of granule after mixing
50be 286 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
Embodiment 17
Obtain inhalant according to embodiment 1-16, carry out pharmacodynamic experiment, wherein drugs compared is HUACHANSU ZHUSHEYE (commercially available), Cutis Bufonis injection A and B, and Cutis Bufonis injection A and B is prepared in accordance with the following methods:
Cutis Bufonis injection A:
Get dry maxima skin to clean, add 15 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2 (85 DEG C), is cooled to 20 DEG C, adds alcohol settling, make alcohol content reach 60%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leaves standstill 48 hours, filters, reclaim ethanol to without alcohol taste, add water to every milliliter and be equivalent to crude drug in whole 5g, regulate pH to 6.5,0.45 micron membrane filter filters, embedding, pressure sterilizing and get final product.
Cutis Bufonis injection B:
Get dry maxima skin to clean, add 20 times amount 60% ethanol, reflux, extract, 2 times, each 2 hours, merge extractive liquid, reclaimed ethanol extremely without alcohol taste, add 3 times of water gagings, deepfreeze 48 hours, filter, add the active carbon (W/V) of 0.3%, boil 30 minutes, filter, add water to every milliliter and be equivalent to crude drug in whole 5g, regulate pH to 7,0.45 micron membrane filter filters, embedding, pressure sterilizing and get final product.
Effect experiment:
The method establishment induced lung animal model for tumour of benzopyrene percutaneous puncture pulmonary injection, gets modeling rat 200, is divided into 20 groups at random, often organizes 10.Every day 1 time, medication 7 days.Administering mode: 1-16 group is the inhalant inhalation of embodiment 1-16, dosage is 15g crude drug/kg; 17th group of lumbar injection HUACHANSU ZHUSHEYE (commercially available), dosage: 15g crude drug/kg; 18th group of lumbar injection Cutis Bufonis injection A, dosage: 15g crude drug/kg; 19th group of lumbar injection Cutis Bufonis injection B, dosage: 15g crude drug/kg; 20th group is saline control group, lumbar injection 10mL/kg.Next day is put to death rat in drug withdrawal, dissects lungs, observes lung lesion situation.
Experimental result: the pulmonary of the rat that inhalant administration is respectively organized occur the incidence rate of lump or nodular lesion minimum be 10.5%, be up to 30.3%; HUACHANSU ZHUSHEYE group (commercially available) is 60.5%, Cutis Bufonis injection A group is 50.1%, Cutis Bufonis injection B group is 60.8%, normal saline group is 100%, and inhalant administration is respectively organized has significant difference (P<0.05) with HUACHANSU ZHUSHEYE group (commercially available), Cutis Bufonis injection A group, Cutis Bufonis injection B group, normal saline group in pulmonary lesion incidence rate.
Embodiment 18
Obtain inhalant according to embodiment 1-16, carry out pharmacodynamic experiment, wherein drugs compared is HUACHANSU ZHUSHEYE (commercially available), Cutis Bufonis injection A and B, and Cutis Bufonis injection A and B is prepared in accordance with the following methods:
Cutis Bufonis injection A:
Get dry maxima skin to clean, add 15 times of water gagings, decoct 2 times, each 2 hours, merging filtrate, being concentrated into relative density is 1.2 (85 DEG C), is cooled to 20 DEG C, adds alcohol settling, make alcohol content reach 60%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 85%, leaves standstill 48 hours, filters, reclaim ethanol to without alcohol taste, add water to every milliliter and be equivalent to crude drug in whole 5g, regulate pH to 6.5,0.45 micron membrane filter filters, embedding, pressure sterilizing and get final product.
Cutis Bufonis injection B:
Get dry maxima skin to clean, add 20 times amount 60% ethanol, reflux, extract, 2 times, each 2 hours, merge extractive liquid, reclaimed ethanol extremely without alcohol taste, add 3 times of water gagings, deepfreeze 48 hours, filter, add the active carbon (W/V) of 0.3%, boil 30 minutes, filter, add water to every milliliter and be equivalent to crude drug in whole 5g, regulate pH to 7,0.45 micron membrane filter filters, embedding, pressure sterilizing and get final product.
Effect experiment:
1, the foundation of rabbit VX2 original position lung cancer model
1) the making conventional recovery VX2 tumor frozen cell suspension of lotus tumor kind rabbit, makes 10
6the living cells suspension of/mL, gets 1mL and is inoculated in intramuscular outside 1 rabbit right rear leg, can to lay one's hand on and a diameter is about the lump of 30cm, namely make lotus tumor kind rabbit after 3 weeks in inoculation position.
2) tissue mass suspension prepare aseptic condition under cut intramuscular tumor outside lotus tumor kind rabbit leg, get in flesh of fish sample eugonic tumor tissues, be cut into pureed.RPMI 1640 culture fluid rinses 30 mesh sieves, removes individual cells and too small piece of tissue, and sieve upper part crosses 20 mesh sieves under RPMI RPMI-1640 rinses, and removes excessive piece of tissue.Get lower part of screen to divide, after centrifugation, taking precipitate 2mL, adds RPMI RPMI-1640 and is mixed with tissue mass suspension 20mL.
3) all rabbit right thoracic wall depilations of inoculation in lung, it is fixing after intramuscular injection Su Mian Xin mixture 0.8mL/kg anaesthetizes, routine disinfection, get aseptic three way cock, connect No. 12 injection needles respectively, suction has the 1mL syringe of rabbit self blood clot and the syringe of Xi You VX2 tumor tissue's suspension or cell suspension 1mL.Under X-ray examination, No. 12 syringe needles are thrust in rabbit inferior lobe of right lung, after resorption depletion of blood, inject 0.4mL tissue mass suspension or cell suspension, then inject 0.2mL rabbit self blood clot, extract syringe needle, inoculate complete.
4) the whole rabbit of the observation of growth of xenografted situation was from after inoculation the 3rd day, and every day carries out to chest the CT scan that layer is wide, thickness is 5mm, observed transfer case in tumor growth, thoracic cavity.
5) rabbit VX2 original position lung cancer model sets up result rabbit all has tumor growth in lung, success ratio of inoculation 100%.
2, effect experiment:
Get successfully modeling rabbit 200, be divided into 20 groups at random, often organize 10.Every day 1 time, medication 12 days.Administering mode: 1-16 group is the inhalant inhalation of embodiment 1-16, dosage: 15g crude drug/kg; 17th group of lumbar injection HUACHANSU ZHUSHEYE (commercially available), dosage: 15g crude drug/kg; 18th group of lumbar injection Cutis Bufonis injection A, dosage: 15g crude drug/kg; 19th group of lumbar injection Cutis Bufonis injection B, dosage: 15g crude drug/kg; 20th group is saline control group, lumbar injection 10mL/kg.Next day is put to death rabbit in drug withdrawal, dissects rabbit lungs, peels off tumor tissues, weighs.
Experimental result: the tumour inhibiting rate of the rabbit that inhalant administration is respectively organized is minimum is 58.7%, is up to 64.3%; HUACHANSU ZHUSHEYE group (commercially available) is 56.3%, Cutis Bufonis injection A group is 45.1%, Cutis Bufonis injection B group is 48.9%, normal saline is to tumor almost unrestraint effect, and inhalant administration is respectively organized has significant difference (P<0.05) with the tumour inhibiting rate of HUACHANSU ZHUSHEYE group (commercially available), Cutis Bufonis injection A group, Cutis Bufonis injection B group, normal saline group.
Claims (1)
1. a toad skin extract dry powder inhalant, is characterized in that it is made up of Cutis Bufonis extract and medically acceptable adjuvant;
Being prepared as of described Cutis Bufonis extract: get Cutis Bufonis and clean, add 10 times of water gagings, decoct 1 hour, at extracting solution is concentrated into 50 DEG C, relative density is 1.20, is cooled to room temperature, adds alcohol settling, make alcohol content reach 50%, leave standstill 24 hours, get supernatant and reclaim ethanol extremely without alcohol taste, concentrated solution adds ethanol makes alcohol content reach 70%, leave standstill 48 hours, filter, reclaim ethanol extremely without alcohol taste, concentrating under reduced pressure, vacuum drying obtains Cutis Bufonis extract;
Being prepared as of described inhalant: by Cutis Bufonis extract comminution by gas stream, powder D
50below 5 microns, this powder is mixed with the lactose granule of 20 times amount again, the D of granule after mixing
50be 286 microns, be packaged in capsule or bubble-cap or Diskus, obtain inhalant.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1803153A (en) * | 2005-09-01 | 2006-07-19 | 安徽威尔曼振星药业有限公司 | Toad essence capsule and its preparation method |
| CN1813802A (en) * | 2005-02-04 | 2006-08-09 | 安徽金蟾生化股份有限公司 | Bufonid skin extract production process |
| CN1857418A (en) * | 2006-04-07 | 2006-11-08 | 南京中医药大学 | Chinese medicine powder inhalant and its preparing method and application |
| CN101396373A (en) * | 2007-09-29 | 2009-04-01 | 四川省中医药科学院 | Cinobufacini extract and preparation method thereof |
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|---|---|---|---|---|
| CN1813802A (en) * | 2005-02-04 | 2006-08-09 | 安徽金蟾生化股份有限公司 | Bufonid skin extract production process |
| CN1803153A (en) * | 2005-09-01 | 2006-07-19 | 安徽威尔曼振星药业有限公司 | Toad essence capsule and its preparation method |
| CN1857418A (en) * | 2006-04-07 | 2006-11-08 | 南京中医药大学 | Chinese medicine powder inhalant and its preparing method and application |
| CN101396373A (en) * | 2007-09-29 | 2009-04-01 | 四川省中医药科学院 | Cinobufacini extract and preparation method thereof |
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