CN103520123B - Cytochrome C freeze-dried powder injection solution - Google Patents
Cytochrome C freeze-dried powder injection solution Download PDFInfo
- Publication number
- CN103520123B CN103520123B CN201310529187.XA CN201310529187A CN103520123B CN 103520123 B CN103520123 B CN 103520123B CN 201310529187 A CN201310529187 A CN 201310529187A CN 103520123 B CN103520123 B CN 103520123B
- Authority
- CN
- China
- Prior art keywords
- cytochrome
- solution
- pharmaceutical composition
- acid
- freeze
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a cytochrome C freeze-dried powder injection solution, and particularly relates to a medicine composite. The cytochrome C freeze-dried powder injection solution comprises cytochrome C, freeze-dried excipients, antioxygen and optional acidifying or alkalizing agents. The cytochrome C freeze-dried powder injection solution can be used for auxiliary treatment of various tissue oxygen lack first aids such as carbon monoxide poisoning, hypnotics poisoning, cyanide poisoning, neonatal asphyxia, oxygen deficit in severe shock stages, cerebrovascular accidents, postconcussional sequelae, breath difficulty caused by anesthesia and pulmonary diseases and treatment of myocardial anoxia caused by various heart diseases.
Description
Technical field
The invention belongs to medical art, relate to a kind of pharmaceutical preparation, particularly relate to a kind of pharmaceutical composition such as lyophilized injectable powder of cytochrome C.Cytochrome C pharmaceutical composition of the present invention can be used for the auxiliary treatment of various histanoxia first aid, the treatment of the myocardial ischemia that the dyspnea caused as carbon monoxide poisoning, hypnotics poisoning, cyanide poisoning, asphyxia of newborn, serious shock phase anoxia, cerebrovas-cularaccident, cerebral concussion sequela, anesthesia and pulmonary disease and various cardiac disorder cause.
Background technology
Cytochrome C (Cytochrome C, Cyt C) or title cytochrome complex (Cytochrome Complex) are a kind of relevant with grain line body inner membrance, loosely organized and small hemoprotein.It is different with other cytochrome, and be a kind of highly soluble albumen, solubility reaches 100 grams often liter.Cytochrome C is the indispensable element of electron transport chain, is responsible for carrying an electronics, transmits between Complex II I and complex IV.In addition, it can accept oxidation and reduction reaction, but is not combined with oxygen.In human body, cytochrome C completes coding by CYCS gene.Cytochrome C is relevant with apoptosis.
The cytochrome C used clinically is a kind of Cellular respiration agonist, and to be present in cell mitochondrial a kind of take iron porphyrin as the protein of prothetic group for it.Cytochrome C is undertaken and is transmitted the effect of electronics, is the indispensable material of Cellular respiration.When histanoxia, exogenous cytochrome C can enter in cell, thus plays its correction Cellular respiration and substance metabolism effect.The protein of this product to be that to be present in cell mitochondrial a kind of with iron porphyrin be prothetic group, at present can purification & isolation.Pharmaceutical formulation is separated from animal hearts or yeast, and cytochrome is a ring of respiratory chain.It is indispensable that various cytochrome rearranges Cellular respiration in certain sequence.Cytochrome C can not permeate through cell membranes, therefore to normal person without effect, when histanoxia, permeability of cell membrane increases, and exogenous preparation can enter in cell, thus plays it and correct Cellular respiration and substance metabolism effect.
Prior art has many preparation methoies about cytochrome C and medicine thereof.Such as, Chinese Patent Application No. 94110756.6 (CN1106019A, Chen Qinggong) disclose a kind of production technology extracting crude cytochrome C from animal hearts, it by Feedstock treating, once extraction, second extraction, absorption, zeolite cleaning, eluting, saltout, precipitate, the processing step such as dialysis completes, it is characterized in that absorbing process adopts the method for layering and precipitating dynamic adsorption, adsorption time can be shortened like this, improve adsorption efficiency, avoid and cause mother solution to go bad because adsorption time is long.
Chinese Patent Application No. 200910229517.7 (CN101709086A, east really) discloses a kind of extracting method of cytochrome C.Take cardiac muscle of mammal as raw material, adopt the preparation of aluminum sulfate solution extraction method, in leaching process, the concentration of aluminum sulfate controls at 0.1-10%, the fixing fabric structure of aluminum sulfate solution is in 1-5 times amount myocardial volume, extracting pH value controls at 3-6, and reaction temperature controls at 0-30 DEG C, and extraction time controls at 1-4h.Again through absorb-elute, finally obtain cytochrome C.The present invention has that method cost is low, step is simple, extraction conditions is gentle, introduce that impurity is few, reactant is pollution-free, be applicable to the feature of industrialized great production.
Chinese Patent Application No. 201110094755.9 (CN102206265A, great achievement) discloses the production method extracting cytochrome c solution with the mammal heart.By mammalian heart through acid extraction, adsorbing material adsorbs, and sour water eluting, saltouts, and precipitation and dialysis obtain cytochrome c solution crude product, and crude product is through ion-exchange purification, and polishing obtains cytochrome c solution.
Chinese Patent Application No. 201110268301.9 (CN102295699A, Northwest University) disclose a kind of new technology for purifying of cytochrome C, comprise the following steps: (1) extracts the crude extract obtaining cytochrome C from animal viscera or yeast; (2) in crude extract, add inorganic salt to saltout, leave standstill the centrifugal foreign protein discarding precipitation, collect supernatant; (3) inorganic salt concentration to 20% ~ 35% in supernatant is regulated with buffer, with forward position hydrophobic chromatography, purification is carried out to it again, be splined on continuously by gained solution in the hydrophobic chromatography post using the inorganic salt solution of 20% ~ 35% equilibrated in advance, collect and penetrate liquid; (4) rinse chromatographic column to absorbance with 20% ~ 35% inorganic salt solution and reach stable, collect chromatograph effluent, and penetrate liquid and merge, be the cytochrome C after purification.The present invention is easy and simple to handle, and separation and purification is effective, and efficiency is high, and cost is low.
Chinese Patent Application No. 200710086421.0 (CN101019836A, Cai Haide) discloses nanometer cytochrome liposome medicine and preparation method thereof.Wherein nanometer cytochrome liposome medicine, comprises the raw material of following parts by weight: neutral phospholipid 40-80; Sitosterol 10-20; Cholesterol 20-40; Cytochrome C 10-30; Polyethylene Glycol 20-25; Reductive glutathione 5-10; Vitamin C 40-50.It is believed that liposome medicament that this invention provides can be used for first aid or the auxiliary treatment of clinical various anoxia.
Chinese Patent Application No. 201210578970.0 (CN103055305A, rich former) discloses the lyophilized formulations of a kind of injection containing cytochrome C pharmaceutical composition, comprises the component of following weight portion: cytochrome C 75 ~ 300 parts; L-arginine 30 ~ 60 parts; Disodium edetate 2 ~ 5 parts; 150 parts, mannitol.This lyophilized formulations selects disodium edetate and L-arginine as adjuvant, be conducive to the stability improving cytochrome C, drug solution preparing process selects mannitol as freeze-dried excipient, selects phosphoric acid or sodium radio-phosphate,P-32 solution as pH value regulator, and product quality can be made more stable; In mannitol solution, after charcoal treatment, add cytochrome C again, avoid the problem of the content reduction that cytochrome C causes because of activated carbon adsorption; The preparation method that this invention adopts, foreshortened to 14 ~ 16 hours by freeze-drying time by 23 hours, reduced energy consumption 30%, simultaneously when not increasing hardware and dropping into, can improve production capacity about 30%, significantly reduce production cost.
Because cytochrome C is a kind of chromoprotein containing ferrum element, it is a very important enzyme in enzymatic oxidising system in organism.Cytochrome C is iron content porphyrin slag conjugated protein, elementary composition by N, S, Fe, C, H five kinds.The Fe content of cytochrome C extracted from pig myocardium is 0.43%, and molecular weight is 13000, isoelectric pH 10.
There is the difficulty that many needs overcome in its Formulation of the macromolecular chemistry structures shape of cytochrome C.The clarity of such as preparation is exactly frequent produced problem.Such as, it is good not that Yu Funian has inquired into cytochron-Sinjection15 clarity in (medical industry, cytochron-Sinjection15 technological innovation, 08 phase in 1980), and inquiring into its main cause is caused by cytochrome C raw material, foreign protein does not eliminate.Again such as, Jiang Hongzhi etc. (medical industry, the improvement of cytochron-Sinjection15 clarity, 10 phases in 1986) are from raising material purity, improved filtering method, controls the aspects such as ingredients pH value and carries out studying to obtaining the cytochron-Sinjection15 with better clarity.
Therefore, provide a kind of cytochrome C product with Good Pharmacy performance such as physical property and/or chemical property and/or biological property, remain that those skilled in the art extremely expect.
Summary of the invention
The object of the present invention is to provide a kind of cytochrome C product of the cytochrome C product with Good Pharmacy performance such as physical property and/or chemical property and/or biological property, particularly a kind of injectable.The present inventor has been surprisingly found that, the cytochrome C pharmaceutical composition with feature of the present invention has excellent pharmaceutical property.The present invention is based on this find and be accomplished.
For this reason, first aspect present invention provides a kind of pharmaceutical composition, wherein comprises: cytochrome C, freeze-dried excipient, antioxidant and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said freeze-dried excipient is selected from mannitol, lactose, sucrose, glucose, sorbitol, Cys, dextran, sodium chloride and combination thereof.The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said freeze-dried excipient is mannitol.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of freeze-dried excipient is 7.5 ~ 150 weight portions, such as 7.5 ~ 100 weight portions, such as 7.5 ~ 75 weight portions.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said antioxidant is selected from sodium sulfite, sodium sulfite and combination thereof.The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said antioxidant is the combination of sodium sulfite and sodium sulfite.The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said antioxidant is that both sodium sulfite and sodium sulfite are with the combination of weight ratio 1:0.5 ~ 2 ratio, being such as with the combination of weight ratio 1:0.75 ~ 1.5 ratio, such as, is with the combination of weight ratio 1:0.8 ~ 1.2 ratio.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of antioxidant (as combinationally use antioxidant then this amount be the summation of whole antioxidant) be 1 ~ 10 weight portion, such as 1 ~ 5 weight portion, such as 2 ~ 3 weight portions.
The pharmaceutical composition of arbitrary embodiment, wherein also optionally comprises Cys according to a first aspect of the present invention.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of Cys is 1 ~ 10 weight portion, such as 1 ~ 5 weight portion, such as 1 ~ 2 weight portion.
The pharmaceutical composition of arbitrary embodiment, wherein also optionally comprises acid-base modifier according to a first aspect of the present invention.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein said acid-base modifier (also can be described as pH adjusting agent in this area) is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulphuric acid or its combination.In one embodiment, described acid-base modifier is hydrochloric acid solution or sodium hydroxide solution, such as 1M hydrochloric acid solution or 1M sodium hydroxide solution.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is dissolved in water and/or dilutes the solution made containing cytochromeC3 mg in every 1ml, the pH value of this solution is 6.0 ~ 7.5, and the pH value of such as this solution is 6.5 ~ 7.5, and the pH value of such as this solution is 7.0 ~ 7.5.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 150 weight portion, antioxidant 1 ~ 10 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 100 weight portion, antioxidant 1 ~ 5 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 75 weight portion, antioxidant 2 ~ 3 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment, wherein also comprises Cys according to a first aspect of the present invention.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 150 weight portion, Cys 1 ~ 10 weight portion, antioxidant 1 ~ 10 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 100 weight portion, Cys 1 ~ 5 weight portion, antioxidant 1 ~ 5 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein comprises: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 75 weight portion, Cys 1 ~ 2 weight portion, antioxidant 2 ~ 3 weight portion and optional acid-base modifier.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is dissolved in water and dilutes the solution made containing cytochromeC3 mg in every 1ml, the pH value of this solution is 6.0 ~ 7.5, and the pH value of such as this solution is 6.5 ~ 7.5, and the pH value of such as this solution is 7.0 ~ 7.5.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is lyophilization injectable powder.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, its solution before lyophilization, except comprising cytochrome C, freeze-dried excipient, antioxidant and optional Cys and optional acid-base modifier, also comprises water for injection.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, its solution before lyophilization also comprises water for injection except comprising cytochrome C, freeze-dried excipient, antioxidant and optional Cys and optional acid-base modifier, the solid content of described solution is 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it redissolves to substantially identical with solution before lyophilization volume with water for injection, solid content in gained solution is 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).
As everyone knows, the lyophilization injectable powder (usually referred to as lyophilized injectable powder or freeze-dried powder) obtained through freezing-vacuum drying, it is first by each material dissolution with solvents (being typically with water dissolution), be mixed with a solution, then this solution is made to carry out freezing, carry out evacuation, distillation again, substantially anhydrous (typically water content is lower than 8% for drying and the one that obtains, particularly be usually less than 5%, be particularly usually less than 3%) Powdered thing or block.Therefore, the acid-base value of this solid lyophilized products regulates the pH value of solution to control by process for preparation usually; Or the pH value that the solid lyophilized products of acquisition can be made under the dissolve/dilute degree of regulation to control this dissolve/dilute liquid by prescription adjustment controls (this is called the acid-base value controlling solid lyophilized products); A rear mode more generally uses usually, such as, in pharmacopeia contained many lyophilized injectable powders control the acid-base value of goods all in this way, and the acid-base value that this mode controls product usually can not the recipe quantity of concrete regulation acid-base modifier, and only specify the acid-base value of finished product.Be equally applicable to of the present inventionly be, pharmaceutical composition according to a first aspect of the present invention described in arbitrary embodiment, the amount of wherein said optional acid-base modifier is, the amount of pH value in 6.0 ~ 7.5 scopes of this solution during the solution making described lyophilized injectable powder water for injection be dissolved into containing active component 3mg/ml concentration, the amount of pH value in 6.5 ~ 7.5 scopes of such as this solution.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein also optionally comprises the excipient being selected from other following kind: mannitol, lactose, sucrose, glucose, sorbitol, Cys, dextran, sodium chloride and combination thereof.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is the pharmaceutical composition of solid type.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is cryodesiccated pharmaceutical composition.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is aseptic freeze-dried injectable powder.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is aseptic freeze-dried injectable powder, its when the preparation of medicinal liquid through activated carbon adsorption processing procedure.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is aseptic freeze-dried injectable powder, and it through activated carbon adsorption processing procedure, with active carbon together with is also added with paper pulp in described activated carbon adsorption process when the preparation of medicinal liquid.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is aseptic freeze-dried injectable powder, its when the preparation of medicinal liquid through activated carbon adsorption processing procedure, this activated carbon adsorption processing procedure is: in the medicinal liquid comprising active component, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, stir 10 ~ 30 minutes.In one embodiment, described paper pulp is prepared with dry filter paper.In one embodiment, the amount of the paper pulp added together with active carbon, in dry filter paper, the concentration in medicinal liquid is 0.05% ~ 0.2% (w/v), preferably 0.05% ~ 0.1% (w/v).
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is aseptic freeze-dried injectable powder, and it obtains according to following methods preparation:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection;
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50-60 DEG C, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, insulated and stirred 10 ~ 30 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, if desired by the scope that acid-base modifier regulates this solution to conform with the regulations to pH;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, the scope that wherein step (5) described pH conforms with the regulations refers to: when this solution water for injection is diluted to the concentration of 3mg/ml, the pH value of gained dilute solution is in 6.0 ~ 7.5 scopes, the pH value of such as this dilute solution is in 6.5 ~ 7.5 scopes, and the pH value of such as this dilute solution is 7.0.Although the activity component concentration of step (b) gained solution measures the normal concentration of pH value higher than finished product, but step (b) solution can easily suitably dilute by those skilled in the art (to the concentration of about 3mg/ml) and measure the pH value of gained diluent, need according to this pH value determining step (b) gained solution the amplitude regulating Acidity of Aikalinity.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, wherein mend to inject water to described in step (5) and refer in right amount and make solid content in this medicinal liquid reach 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).Thus, be dissolved in appropriate water for injection described in step (1) by each solid adjuvant material and cytochrome C, this appropriate water for injection makes wherein solid concentration higher than the middle liquid solid concentration of step (5).
Have been found that when carrying out charcoal absorption, the injection adding appropriate paper pulp acquisition has better physical stability in storage, and particularly particulate matter change (increase) is not obvious.
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, the amount of the paper pulp added in wherein said step (3), in dry filter paper, the concentration in medicinal liquid is 0.05% ~ 0.2% (w/v), preferably 0.05% ~ 0.1% (w/v).The occupation mode of the paper pulp when preparing injection as well known to those skilled in the art, such as, can prepare paper pulp according to under type: get common large filter paper and tear to bits, soak in good time, be broken into atherosclerotic with bruisher with appropriate water for injection.This area also has other to prepare the method for paper pulp, such as, in the recording high document of Hou Yu (Hou Yusong, etc., Chinese patent medicine, the 19th volume the 4th phase page 4 in 1997).The concentration of paper pulp can need to determine according to use usually, usually can be such as 10% ~ 50% broad range in.Literary composition is prepared in the technique of injection under the invention, and if not otherwise indicated, paper pulp used is the paper pulp (being equivalent to the slurry that the dry filter paper water for injection of 30g makes 100ml in right amount) of use 30%; The amount of the paper pulp added in step (2), be converted to dry filter paper and calculate, dry filter paper concentration is in the solution 0.05% ~ 0.2% (w/v), preferably 0.05% ~ 0.1% (w/v).
The pharmaceutical composition of arbitrary embodiment according to a first aspect of the present invention, it is lyophilization injectable powder, it checks according to " first method (light blockage method) " in Pharmacopoeia of People's Republic of China version in 2010 two (in the present invention can referred to as " Chinese Pharmacopoeia 2010 editions two " or similar address) annex IX C particulate matter inspection technique, particle number containing >=10 μm in each test sample container is less than 6000, such as be less than 3000, such as be less than 1500, such as be less than 1000, such as be less than 500, such as, be less than 250; Such as, such as, such as, such as, such as, and/or the particle number containing >=25 μm in each test sample container is less than 600, is less than 300, is less than 150, be less than 100, be less than 50, be less than 25.
The present invention has been surprisingly found that, in adding in the compositions of the present invention when the appropriate mannitol as excipient and Cys, can give the preparation performance that lyophilization powder injection composition of the present invention is good.
Further, second aspect present invention provides a kind of method preparing cytochrome C pharmaceutical composition such as lyophilization injectable powder described in the arbitrary embodiment of cytochrome C pharmaceutical composition such as first aspect present invention, comprises: cytochrome C, freeze-dried excipient, antioxidant and optional acid-base modifier in described cytochrome C pharmaceutical composition; The method comprises the following steps:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection;
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50-60 DEG C, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, insulated and stirred 10 ~ 30 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, if desired by the scope that acid-base modifier regulates this solution to conform with the regulations to pH;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.
The method of arbitrary embodiment according to a second aspect of the present invention, the scope that wherein step (5) described pH conforms with the regulations refers to: when this solution water for injection is diluted to the concentration of 3mg/ml, the pH value of gained dilute solution is in 6.0 ~ 7.5 scopes, the pH value of such as this dilute solution is in 6.5 ~ 7.5 scopes, and the pH value of such as this dilute solution is 7.0.Although the activity component concentration of step (b) gained solution measures the normal concentration of pH value higher than finished product, but step (b) solution can easily suitably dilute by those skilled in the art (to the concentration of about 3mg/ml) and measure the pH value of gained diluent, need according to this pH value determining step (b) gained solution the amplitude regulating Acidity of Aikalinity.
The method of arbitrary embodiment according to a second aspect of the present invention, wherein mend to inject water to described in step (5) and refer in right amount and make solid content in this medicinal liquid reach 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).Thus, be dissolved in appropriate water for injection described in step (1) by each solid adjuvant material and cytochrome C, this appropriate water for injection makes wherein solid concentration higher than the middle liquid solid concentration of step (5).
Method according to a second aspect of the present invention described in arbitrary embodiment, wherein said acid-base modifier is the aqueous solution being selected from following acid-base modifier: sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulphuric acid or its combination.The concentration of these aqueous solutions well known to a person skilled in the art, such as 1 ~ 10%, such as 2% ~ 5%.In one embodiment, described acid-base modifier is hydrochloric acid solution or sodium hydroxide solution, such as 1M hydrochloric acid solution or 1M sodium hydroxide solution.
Method according to a second aspect of the present invention described in arbitrary embodiment, wherein in step (6) after removing moisture in gained lyophilization material moisture lower than 10%, preferably lower than 8%, preferably lower than 5%, more preferably less than 3%.
Method according to a second aspect of the present invention described in arbitrary embodiment, wherein said paper pulp is prepared with dry filter paper.In one embodiment, the amount of the paper pulp added together with active carbon, in dry filter paper, the concentration in medicinal liquid is 0.05% ~ 0.2% (w/v), preferably 0.05% ~ 0.1% (w/v).
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 150 weight portion, antioxidant 1 ~ 10 weight portion and optional acid-base modifier.
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 100 weight portion, antioxidant 1 ~ 5 weight portion and optional acid-base modifier.
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 75 weight portion, antioxidant 2 ~ 3 weight portion and optional acid-base modifier.
The method of arbitrary embodiment according to a second aspect of the present invention, also comprises Cys in wherein said pharmaceutical composition.
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 150 weight portion, Cys 1 ~ 10 weight portion, antioxidant 1 ~ 10 weight portion and optional acid-base modifier.
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 100 weight portion, Cys 1 ~ 5 weight portion, antioxidant 1 ~ 5 weight portion and optional acid-base modifier.
The method of arbitrary embodiment according to a second aspect of the present invention, comprises in wherein said pharmaceutical composition: cytochrome C 7.5 weight portion, freeze-dried excipient 7.5 ~ 75 weight portion, Cys 1 ~ 2 weight portion, antioxidant 2 ~ 3 weight portion and optional acid-base modifier.
Further, third aspect present invention provide cytochrome C pharmaceutical composition described in the arbitrary embodiment of first aspect present invention or the cytochrome C pharmaceutical composition prepared by the arbitrary embodiment of second aspect present invention for the preparation of various histanoxia first aid and auxiliary treatment, as carbon monoxide poisoning, hypnotics poisoning, cyanide poisoning, asphyxia of newborn, serious shock phase anoxia, cerebrovas-cularaccident, cerebral concussion sequela, purposes in the medicine of the treatment of the dyspnea that anesthesia and pulmonary disease cause and the myocardial ischemia that various cardiac disorder causes.
State on the invention in the step of preparation method, although its concrete steps described in some details or the language step described in preparation example that describes up and down literary composition detailed description of the invention part distinguish to some extent, but those skilled in the art can summarize the above method step completely according to the open in detail of the present invention's full text.
Arbitrary embodiment of either side of the present invention, can combine with other embodiment, as long as they there will not be contradiction.In addition, in arbitrary embodiment of either side of the present invention, arbitrary technical characteristic goes for this technical characteristic in other embodiment, as long as they there will not be contradiction.The invention will be further described below.
All documents that the present invention quotes from, their full content is incorporated to herein by reference, and if the implication expressed by these documents and the present invention inconsistent time, be as the criterion with statement of the present invention.In addition, the various term that the present invention uses and phrase have and well known to a person skilled in the art general sense, nonetheless, the present invention still wishes to be described in more detail at this these terms and phrase and to explain, the term mentioned and phrase, if any inconsistent with common art-recognized meanings, are as the criterion with the implication that the present invention states.
As above hereinafter described, have been surprisingly found that, in dosing process, use active carbon and paper pulp combination, can make the composition of liquid medicine of acquisition such as injection have good physical stability.
The pharmaceutical composition comprising cytochrome C of the present invention, although be provide with the injectable powder of solid type in certain embodiments, but be in some embodiments, medicinal liquid directly can also be dispensed in vial without lyophilization process by the injectable powder compositions of these solid types and make aqueous injection.The preparation method of the aqueous injection of solution-type well known to a person skilled in the art.
[assay]
This determination is used for the present invention, can measure the content of the various cytochrome C comprised in the sample of cytochrome C:
The preparation of need testing solution: precision measures this product appropriate (being about equivalent to cytochrome C1 00mg), puts in 10ml measuring bottle, is diluted with water to scale;
Precision measures need testing solution 1ml, put in 50ml measuring bottle, (sodium dihydrogen phosphate 1.38g and sodium hydrogen phosphate 31.2g is got with phosphate buffer, add water and make in right amount to be dissolved into 1000ml, adjust ph to 7.3) be diluted to scale, add sodium dithionite and be about 15mg, shake up, according to ultraviolet visible spectrophotometry (Chinese Pharmacopoeia version in 2010 two annex IVA), at the wavelength place of about 550nm, find out maximum absorption wavelength with interval 0.5nm, measure absorbance, by the absorptance of cytochrome C
be the content (mg/ml) of cytochrome C in 23.0 calculating test samples.
[vitality test]
This vitality test method is used for the present invention, can measure the various cytochrome C vigor comprising the sample of cytochrome C:
(1) phosphate buffer (0.2mol/L): get phosphoric acid hydrogen two and receive 71.64g, add water and make to be dissolved into 1000ml, as first liquid.Separately get sodium dihydrogen phosphate 27.60g, add water and make to be dissolved into 1000ml, as second liquid.Get first liquid 81ml and second liquid 19ml, mixing, adjust ph to 7.3;
(2) phosphate buffer (0.lmol/L): get phosphate buffer (0.2mol/L) 500ml, be diluted with water to 1000ml, adjust ph to 7.3;
(3) phosphate buffer (0.02mol/L): get phosphate buffer (0.2mol/L) 100ml, be diluted with water to 1000ml, adjust ph to 7.3;
(4) succinate solution: get succinic acid and each 4.72g of potassium hydroxide, add water and make to be dissolved into 100ml, adjust ph to 7.3;
(5) potassium cyanide solution: get potassium cyanide 0.65g, adds water after making to be dissolved into 100ml, by dilute sulfuric acid adjust ph to 7.3;
(6) Suspension of cytochrome C is removed: get 2, fresh pig (cattle) heart, except degrease and connective tissue, cut into inch strips, rub with meat grinder, put in gauze pocket, within about 2 hours, (often stir with tap water, extrude hematochrome), extract, wash with water for several times, extract, put in phosphate buffer (0.lmol/L) and soak about 1 hour, extract, repeat immersion 1 time, wash with water for several times, extract, put in tissue mashing machine, add phosphate buffer (0.02mol/L) and just make the submergence of pig (cattle) heart in right amount, be pounded homogenate, centrifugal 10 minutes (generic centrifuge), get upper strata suspension, on a small quantity on the rocks, rapid spirit of vinegar adjust ph is to about 5.5, centrifugal 15 minutes immediately, get precipitation, add isopyknic phosphate buffer (0.lmol/L), after even with glass homogenizer mill, be stored in refrigerator, face the used time and get 1.0ml, add phosphate buffer (0.lmol/L) and be diluted to 10ml,
(7) preparation of need testing solution: get test sample, add water the solution made and be about 3mg in every lml containing cytochrome C;
(8) algoscopy: get phosphate buffer (0.2mol/L) 5ml, succinate solution 1.0ml and need testing solution 0.5ml, put in 25ml color-comparison tube, add Suspension 0.5ml and the potassium cyanide solution 1.0ml of cytochrome C, be diluted with water to 10ml, shake up, do blank with same reagent, according to ultraviolet visible spectrophotometry (Chinese Pharmacopoeia version in 2010 two annex IV A), near the wavelength place of 550nm, interval 0.5min finds out maximum absorption wavelength, and measure absorbance, till absorbance no longer increases, as enzyme reduction absorbance, then respectively add sodium dithionite and be about 5mg, shake up, place about 10 minutes, measure absorbance, till absorbance no longer increases, as electronation absorbance at above-mentioned Same Wavelength place, be calculated as follows cytochrome C vigor:
Cytochrome C pharmaceutical composition provided by the invention, its active component is cytochrome C, can clinically for the auxiliary treatment of various histanoxia first aid, the treatment of the myocardial ischemia that the dyspnea caused as carbon monoxide poisoning, hypnotics poisoning, cyanide poisoning, asphyxia of newborn, serious shock phase anoxia, cerebrovas-cularaccident, cerebral concussion sequela, anesthesia and pulmonary disease and various cardiac disorder cause.
Cytochrome C pharmaceutical composition provided by the invention is as the injectable powder of solid type, and usually can be made into every bottled cytochrome C amount is the specification of 15mg.If make the aqueous injection of solution-type, the dosage form that specification is 2ml:15mg usually can be made.
Clinically, the usage and dosage of cytochrome C pharmaceutical composition provided by the invention can be: adult: intravenous injection or instillation, a 15 ~ 30mg, every day 30 ~ 60mg.During intravenous injection, add after 25% glucose injection 20ml mixes and slowly inject.Also can instil with 5 ~ 10% glucose injections or 0.9% sodium chloride injection dilution posterior vein.Child: intramuscular injection: <1 year, each 1.5 ~ 7.5mg; 1 ~ 8 year old, each 15mg; 9 years old, each 15 ~ 30mg, every day 1 time.Quiet note: <11 year, each 7.5mg; 1 ~ 8 year old, each 7.5 ~ 15mg; >9 year, each 15 ~ 30mg, every day 1 time; Quiet: <l8 year, each 15mg; >9 year, each 15 ~ 30mg, every day 1 time.
Typically, need to do hypersensitive test before cytochrome C pharmaceutical composition provided by the invention medication, the skin test cut genealogy of law 0.03% solution 1, drip and bend on surface skin in forearm, with pin thereon puncture once (single thorn) or many under (thorniness), at least measure extent of hemorrhage.Intradermal injection genealogy of law 0.03mg/ml solution 0.03 ~ 0.05ml intradermal injection.All observe 20 minutes, single thorn person local blush more than diameter 10mm or pimple more than diameter >7mm, thorniness and intradermal injection person's blush more than diameter 15mm or pimple more than diameter 10mm are positive.Skin test positive person is forbidden.When continuing medication again after stopping medication, anaphylaxis especially easily occurs, and must do skin test again, and the intracutaneous injection that application dosage is less.
In cytochrome C pharmaceutical composition provided by the invention, its active component cytochrome C is Cellular respiration agonist, and to be present in cell mitochondrial a kind of take iron porphyrin as the protein of prothetic group for it.Cytochrome C is undertaken and is transmitted the effect of electronics, is the indispensable material of Cellular respiration.When histanoxia, exogenous cytochrome C can enter in cell, thus plays its correction Cellular respiration and substance metabolism effect.The protein of this product to be that to be present in cell mitochondrial a kind of with iron porphyrin be prothetic group, at present can purification & isolation.Pharmaceutical formulation is separated from animal hearts or yeast, and cytochrome is a ring of respiratory chain.It is indispensable that various cytochrome rearranges Cellular respiration in certain sequence.Cytochrome C can not permeate through cell membranes, therefore to normal person without effect, when histanoxia, permeability of cell membrane increases, and exogenous preparation can enter in cell, thus plays it and correct Cellular respiration and substance metabolism effect.
As the cytochrome C pharmaceutical composition of the present invention of injectable powder type, the normally peach lyophilizing block of its character.As the cytochrome C pharmaceutical composition of the present invention of solution-type, its character can be orange-red clear liquid usually.The common holding conditions of these cytochrome C pharmaceutical compositions of the present invention is: airtight, preserves at cool dark place (lucifuge is no greater than 20 DEG C), and preserving usual effect duration with this understanding can reach 24 months.
According to the present invention, term " excipient " also can be described as adjuvant, filler etc." the acceptable excipient of pharmacy " used herein refers to the excipient that can be used for compounding pharmaceutical, it there is no harmful effect to organism, and normally organism can tolerate.
The preparation process of lyophilization injectable powder well known to a person skilled in the art pharmaceutical technology, such as following kind of the schematic freeze-drying curve of two shown in freeze-drying curve A and freeze-drying curve B:
Hereafter preparing in the instantiation in lyophilization injectable powder, if not otherwise specified, freeze-drying curve used is freeze-drying curve A.
Moisture in lyophilization injectable powder is general below 8%, preferably lower than 5%, more preferably less than 3%.Moisture Control is by suitably adjusting lyophilization program to control.Moisture in this lyophilization injectable powder can measure according to many known methods, such as dry weight-loss method.
In the present invention, in order to regulate the pH value of medicinal liquid where necessary, suitable pH adjusting agent can be added in compositions.Although the present inventor only regulates with not having the strong acid of buffer capacity or strong base solution such as a sodium hydrate aqueous solution and aqueous hydrochloric acid solution, but, those skilled in the art understand, if the pH requirement of system can be met with this pH adjusting agent process of not having buffer capacity, the pH adjusting agent then with buffer capacity will can realize the object of the invention more, therefore these buffer agents not only can adjust ph, and can stablize pH value.Therefore arbitrary pH adjusting agent listed by the present invention or its combination include in spirit and scope of the invention.
When preparing lyophilized injectable powder of the present invention, in the medicinal liquid prepared, solid content is 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).Obtain because lyophilized injectable powder normally carries out lyophilization in tubulose cillin bottle, those skilled in the art understand this product at acquisition finished product even before for doctor, usually a round pie is all presented, although in the volume theory of this cake, lecture is fewer than the volume of original aqueous solution (slightly reducing), but this reducing can not narrow down to former aqueous solution volume 50% usually usually, usual meeting is between the 80-120% of former aqueous solution volume, between the 90-100% being more typically in former aqueous solution volume, and can be observed in finished product cillin bottle former aqueous solution liquid level vestige (main body cake because of lyophilizing reduce after remain in liquid level vestige bottle wall, even if the dried frozen aquatic products in cillin bottle is Powdered because of reasons such as a variety of causes such as collide, usually original liquid level vestige can still be retained), vestige also can estimate the aqueous solution volume of this freeze-dried composition before lyophilization accordingly.Therefore, although the present invention is to provide a kind of substantially anhydrous lyophilization injectable powder, but still roughly can estimate it when preparing according to this injectable powder, medicine liquid volume at least before lyophilization starts, the weight of the dry end-product in the volume estimated according to this and cillin bottle, also can calculate when preparing lyophilized injectable powder of the present invention, the content of the solid content in the medicinal liquid prepared.Therefore, lyophilized injectable powder according to a first aspect of the present invention, its solid content of medicinal liquid when preparing is 1.5 ~ 20% (w/v), such as 2 ~ 15% (w/v), such as 2 ~ 12% (w/v).
Term " solid content " refers to solid matter (such as reactive compound of the present invention and whole excipient used, weight/gram) join in solvent (such as water for injection), a solution is obtained after dissolving, the weight of described solid matter such as, divided by the percent (weight/volume percent, g/100ml) of whole liquor capacity.Such as in the present invention, add appropriate aqueous solution for injection with 100mg reactive compound and other solid content amounting to about 50mg, be mixed with the solution that final volume is 1ml, its solid content is 15%.
In the present invention, symbol %, according to the linguistic context that it uses, can have the implication of those skilled in the art's easy understand.Such as when mentioning solid content, this symbol represents the percent (w/v, such as g/100ml) of weight/volume; Again such as when mentioning " water content " in lyophilization injectable powder, such as water content is below 8%, and now this symbol % represents the percent (w/w, g/100g) of w/w.Generally speaking, solid dispersal in a liquid time, % represents weight/volume percent; Solid dispersal in solids or liquid dispersion in solids (such as the water content of powder pin) time, % represents w/w percent.In other cases, unless otherwise noted, symbol % represents w/w percent.
When preparing medicinal liquid of the present invention, as well known to those skilled in the art, the microporous filter membrane of example 0.45um according to appointment can carry out coarse filtration filtration, by before in liquid medicine filling to cillin bottle, the microporous filter membrane of example 0.22um according to appointment can carry out fine straining and filter with degerming, can filter repeatedly if desired.
According to pharmaceutical composition of the present invention, it is lyophilization injectable powder.In one embodiment, this lyophilization injectable powder is single-dose preparations (injectable powder that such as XiLin is bottled), in per unit dosage, the amount (it is all converted in the present invention if not otherwise indicated in cytochrome C) of reactive compound can such as but not limited to about 7.5mg, about 15mg, about 30mg, about 50mg.
According to pharmaceutical composition of the present invention, it redissolves with water for injection, and typically the redissolution time is in 30 seconds, preferably in 20 seconds, more preferably in 15 seconds.
According to lyophilized injectable powder of the present invention, it is made in every 1ml containing the solution of reactive compound 3mg and according to the method mensuration under Chinese Pharmacopoeia version in 2010 two annex VI H items with water, the pH value of this solution is 6.0 ~ 7.5, the pH value of such as this solution is 6.5 ~ 7.5, and the pH value of such as this solution is 7.0 ~ 7.5.
Lyophilized injectable powder provided by the invention can be preserved at least 24 months in cool dark place, can meet the Storage Requirement of general lyophilization injectable powder.
Have been found that cytochrome C pharmaceutical composition of the present invention has good pharmaceutical properties.
Detailed description of the invention
Can be conducted further description the present invention by the following examples, but scope of the present invention is not limited to following embodiment.One of skill in the art can understand, and under the prerequisite not deviating from the spirit and scope of the present invention, can carry out various change and modification to the present invention.The present invention carries out generality and/or concrete description to the material used in test and test method.Although for realizing many materials that the object of the invention uses and operational approach is well known in the art, the present invention still describes in detail as far as possible at this.Following examples further illustrate the present invention, instead of restriction the present invention.In example below, the pH adjusting agent (in the present invention that is acid-base modifier) used, unless otherwise noted, 1M sodium hydroxide solution or 1M hydrochloric acid solution, its consumption is when the injectable powder of preparation (can also be applicable to aqueous injection) water for injection being dissolved and/or is diluted to containing active component 3mg/ml solution, reaches value or the scope of defined in following example.
The hereafter object of preparation process in order to illustrate, and based on each citing comparability and make some specific description, those skilled in the art therefrom can summarize the method obtaining the present invention and prepare lyophilized injectable powder or aqueous injection completely according to existing knowledge.Dosing is prepared in various compositions below, and if not otherwise indicated, the total dosing amount often criticized is 1000ml.But when listing formula and preparation process, for injectable powder or aqueous injection, illustrate formula and method for making with the composition of other material of every 7.5mg part cytochrome C and corresponding weight portion.No matter be liquid drugs injection or powder pin, when subpackage, every bottle is 15mg containing active component cytochrome C.
In addition, the method that the particulate matter of aqueous injection or injectable powder checks is carried out according to " first method (light blockage method) " in Chinese Pharmacopoeia 2010 editions two annex IX C particulate matter inspection techniques.
embodiment 1: prepare cytochrome C powder pin
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.75ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 55 DEG C, add 0.25% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.075% (w/v)), insulated and stirred 20 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stir, measure solution ph and optional mensuration active component content, regulate this solution to pH value 7.0 (this pH value is that this solution water for injection is diluted to value measured when activity component concentration is 3mg/ml, lower same) with acid-base modifier if desired;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture (making moisture lower than 3%, lower same), tamponade, to obtain final product.The numbering of this sample is designated as Ex1.
embodiment 2: prepare cytochrome C powder pin
| cytochrome C | 7.5mg, |
| mannitol | 7.5mg, |
| sodium sulfite | 1.2mg, |
| sodium sulfite | 1mg, |
| water for injection | in right amount, 0.85ml is added to. |
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.7ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 60 DEG C, add 0.05% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.1% (w/v)), insulated and stirred 10 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, regulates this solution to pH value 7.5 if desired with acid-base modifier;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.The numbering of this sample is designated as Ex2.
embodiment 3: prepare cytochrome C powder pin
| Cytochrome C | 7.5mg， |
| mannitol | 75mg, |
| sodium sulfite | 0.8mg, |
| sodium sulfite | 1mg, |
| water for injection | in right amount, 0.7ml is added to. |
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.5ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50 DEG C, add 0.5% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.05% (w/v)), insulated and stirred 30 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, regulates this solution to pH value 6.5 if desired with acid-base modifier;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.The numbering of this sample is designated as Ex3.
additional embodiment 1:
With reference to formula and the method for making of embodiment 1, different is only in step (3), the amount of paper pulp is used to be changed, change 0,0.01%, 0.02%, 0.04%, 0.05%, 0.08%, 0.1%, 0.125%, 0.15%, 0.25%, 0.5% (w/v) respectively into, obtain 11 injectable powder formula, its numbering is designated as Ex101, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex108, Ex109, Ex110, Ex111 respectively;
With reference to formula and the method for making of embodiment 1, different is only in step (3), do not use active carbon, and the amount of paper pulp changes 0.05%, 0.1%, 0.25%, 0.5%, 1.0% (w/v) respectively into, obtain 5 injectable powder formula, its numbering is designated as Ex121, Ex122, Ex123, Ex124, Ex125 respectively;
With reference to formula and the method for making of embodiment 2, different is only in step (3), the amount of paper pulp is used to be changed, change 0,0.02%, 0.04%, 0.05%, 0.08%, 0.1%, 0.125%, 0.25%, 0.5% (w/v) respectively into, obtain 9 injectable powder formula, its numbering is designated as Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex137, Ex138, Ex139 respectively;
With reference to formula and the method for making of embodiment 3, different is only in step (3), the amount of paper pulp is used to be changed, change 0,0.02%, 0.04%, 0.05%, 0.08%, 0.1%, 0.125%, 0.25%, 0.5% (w/v) respectively into, obtain 9 injectable powder formula, its numbering is designated as Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex147, Ex148, Ex149 respectively;
Adopt following formula: Cytochrome C (i.e. cytochrome C) 15mg, glucose 15mg, sodium sulfite 2.5mg, sodium sulfite 2.5mg, water for injection 0.7ml; Preparation method is with reference to embodiment 1, but adding paper pulp amount in the concentration of dry filter paper in medicinal liquid in step (3) is 0,0.02%, 0.05%, 0.08%, 0.1%, 0.15%, 0.5% (w/v), obtain 7 injectable powder formula, its numbering is designated as Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157 respectively;
With reference to formula and the method for making of embodiment 1, different is only that mannitol is wherein replaced with equivalent: lactose, sucrose, glucose, sorbitol, dextran, sodium chloride, obtain 6 injectable powder formula, its numbering is designated as Ex161, Ex162, Ex163, Ex164, Ex165, Ex166 respectively.
test 1: the pharmaceutical property investigating each compositions sample
Sample: each sample Ex1 of embodiment 1 ~ embodiment 3, Ex2, Ex3, each sample Ex101 of additional embodiment 1, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex108, Ex109, Ex110, Ex111, Ex121, Ex122, Ex123, Ex124, Ex125, Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex137, Ex138, Ex139, Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex147, Ex148, Ex149, Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157, Ex161, Ex162, Ex163, Ex164, Ex165, Ex166.
Method: each injectable powder sample is placed in 35 DEG C of dark places and places 6 months (in the present invention can referred to as disposing in high-temperature treatment or 35 DEG C-June or 35 DEG C-June), concurrently each injectable powder sample is placed in 5 DEG C of dark places to place 6 months (can dispose referred to as low temperature in the present invention or 5 DEG C-June or 5 DEG C-June are disposed) simultaneously, check the particulate matter of each sample when June, add up each sample >=10 μm of particle numbers.For each sample, calculate the percent change of >=10 μm of particle numbers after above-mentioned high-temperature process, this parameter is called microgranule percent change, and calculating formula is as follows:
Wherein, after high-temperature treatment after >=10 μm of particle numbers and low temperature are disposed >=10 μm of particle numbers are that sample obtains according to light blockage method assay method mentioned above.Above-mentioned microgranule percent change (%) more shows more stable close to 100%, increase higher than 100% particulate matter, and product is unstable, is worth more high more unstable.
Result: for Ex1, Ex2, Ex3, Ex105, Ex106, Ex107, Ex134, Ex135, Ex136, Ex144, Ex145, the sample of these paper pulp consumptions 0.05% ~ 0.1% of Ex146, and Ex108, Ex109, Ex110, Ex111, Ex137, Ex138, Ex139, Ex147, Ex148, the sample that these paper pulp consumptions of Ex149 are greater than 0.1%, their microgranule percent change (%) is all in 93 ~ 126% scopes, such as Ex1, Ex2, the microgranule percent change (%) of Ex3 three is respectively 112%, 104%, 96%, show very gratifying physical stability,
For Ex101, Ex102, Ex103, Ex104, Ex131, Ex132, Ex133, with these paper pulp consumptions of Ex141, Ex142, Ex143 lower than 0.05% sample, their microgranule percent change (%) is all in 179 ~ 297% scopes, the microgranule percent change (%) of such as both Ex101, Ex103 is respectively 234%, 287%, and display physical stability cannot make us accepting completely;
For Ex121, Ex122, Ex123, Ex124, Ex125, these only do not use the sample of active carbon (although paper pulp and active carbon equally usual be yet the adsorbent of injection) with paper pulp, their microgranule percent change (%) is all in 227 ~ 357% scopes, the microgranule percent change (%) of such as both Ex121, Ex123 is respectively 302%, 281%, and display physical stability cannot make us accepting completely;
For Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157, these change mannitol into glucose and adopt the sample of not commensurability paper pulp, and mannitol to be changed into the sample of other conventional injectable powder excipient by these for Ex161, Ex162, Ex163, Ex164, Ex165, Ex166, their microgranule percent change (%) is all in 193 ~ 366% scopes, the microgranule percent change (%) of such as both Ex151, Ex165 is respectively 288%, 258%, and display physical stability cannot make us accepting completely;
Above result display, the paper pulp adding more than 0.05% amount is extremely useful for improving the physical stability of product in follow-up Long-term Storage process, although paper pulp this be only when preparing for adsorbing contaminant usually used as the adsorbent in injection process for preparation, and prior art there is no, and any about this common adsorbents of paper pulp, it can contribute to improving the instruction of injection physical stability.But this result improving physical stability only occurs over just with mannitol when being injectable powder excipient, have no this technique effect when using other excipient.
test 2: investigate the active ingredient loss of each compositions sample in process for preparation
Sample: each sample Ex1 of embodiment 1 ~ embodiment 3, Ex2, Ex3, each sample Ex101 of additional embodiment 1, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex108, Ex109, Ex110, Ex111, Ex121, Ex122, Ex123, Ex124, Ex125, Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex137, Ex138, Ex139, Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex147, Ex148, Ex149, Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157.
Calculate the active ingredient loss of each sample in process for preparation of above embodiment 1 ~ embodiment 3 and additional embodiment 1, computational methods are, determine the amount (W1) of active component in raw material inventory, measure and in calculation procedure (5) after the microporous filter membrane aseptic filtration of 0.22 μm the amount (W2) of contained active component in the whole medicinal liquid of gained, be calculated as follows active ingredient loss (%):
Active ingredient loss (%)=[(W1-W2) ÷ W1] × 100%
Known in art technology people; for the preparation of injectable powder; usually dosing loss is had in charcoal adsorption process; this loss is disadvantageous for the more expensive medicine of this price of cytochrome C, and above-mentioned active ingredient loss (%) can reflect/evaluate this loss in product configuration process effectively.
Result: Ex1, Ex2, Ex3, Ex101, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex121, Ex122, Ex123, Ex124, Ex125, Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex151, Ex152, Ex153, Ex154, the sample that these paper pulp consumptions of Ex155 are lower or only do not use the sample of active carbon with paper pulp, their active ingredient loss (%) is all in 0.19% ~ 2.57% scope, such as Ex1, Ex2, the active ingredient loss (%) of Ex3 three is respectively 0.37%, 1.92%, 0.86%.But surprisingly, for Ex108, Ex109, Ex110, Ex111, Ex137, Ex138, Ex139, the higher active ingredient loss of sample in process for preparation (%) of these paper pulp consumptions of Ex147, Ex148, Ex149, Ex156, Ex157 is all in 11.2% ~ 19.6% scope, and the active ingredient loss (%) of such as both Ex108, Ex110 is respectively 13.6%, 16.7%.
Visible according to above result, in the dosing process of step (3), use 0.05 ~ 0.1% paper pulp to be useful, consumption is too high is to cause the dosing of active component to lose, and the compositions obtained when consumption is too low physical stability in Long-term Storage process is undesirable.
embodiment 4: prepare cytochrome C injectable powder
| cytochrome C | 7.5mg, |
| mannitol | 35mg, |
| cys | 1.2mg, |
| sodium sulfite | 1.25mg, |
| sodium sulfite | 1.25mg, |
| water for injection | in right amount, 1ml is added to. |
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.75ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 60 DEG C, add 0.05% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.1% (w/v)), insulated and stirred 10 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, regulates this solution to pH value 7.5 if desired with acid-base modifier;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.The numbering of this sample is designated as Ex4.
embodiment 5: prepare cytochrome C injectable powder
| cytochrome C | 7.5mg, |
| mannitol | 10mg, |
| cys | 1mg, |
| sodium sulfite | 2mg, |
| sodium sulfite | 1mg, |
| water for injection | in right amount, 1ml is added to. |
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.75ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50 DEG C, add 0.5% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.05% (w/v)), insulated and stirred 30 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, regulates this solution to pH value 6.5 if desired with acid-base modifier;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.The numbering of this sample is designated as Ex5.
embodiment 6: prepare cytochrome C injectable powder
| Cytochrome C | 7.5mg， |
| Mannitol | 75mg， |
| Cys | 2mg， |
| Sodium sulfite | 1mg， |
| Sodium sulfite | 1mg， |
| water for injection | in right amount, 0.75ml is added to. |
Method for making:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection and (make liquor capacity reach about 0.6ml);
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 55 DEG C, add 0.25% (w/v) active carbon, and add paper pulp and (prepare with dry filter paper, the amount of the paper pulp added, in dry filter paper, the concentration in medicinal liquid is 0.075% (w/v)), insulated and stirred 20 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, regulates this solution to pH value 7.0 if desired with acid-base modifier;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.The numbering of this sample is designated as Ex6.
additional embodiment 2:
With reference to formula and the method for making of embodiment 4, different is only change Cys consumption wherein into 0mg, 0.25mg, 0.5mg, 0.75mg, 1.0mg, 1.25mg, 1.5mg, 2mg, 2.5mg, 5mg, obtains its numbering of 10 injectable powder formula and is designated as Ex201, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex209, Ex210 respectively;
With reference to formula and the method for making of embodiment 4, different is only change Cys wherein into D-Cys 0.75mg, 1.0mg, 1.5mg, 2mg, 2.5mg, obtains its numbering of 5 injectable powder formula and is designated as Ex211, Ex212, Ex213, Ex214, Ex215 respectively; And change Cys wherein into DL-cysteine 0.75mg, 1.0mg, 1.5mg, 2mg, 2.5mg, obtain its numbering of 5 injectable powder formula and be designated as Ex216, Ex217, Ex218, Ex219, Ex220 respectively;
With reference to formula and the method for making of embodiment 5, different is only change Cys consumption wherein into 0mg, 0.5mg, 1.0mg, 1.5mg, 2mg, 3mg, 5mg, obtains its numbering of 7 injectable powder formula and is designated as Ex221, Ex222, Ex223, Ex224, Ex225, Ex226, Ex227 respectively;
With reference to formula and the method for making of embodiment 6, different is only change Cys consumption wherein into 0mg, 0.5mg, 1.0mg, 1.5mg, 2mg, 3mg, 5mg, obtains its numbering of 7 injectable powder formula and is designated as Ex231, Ex232, Ex233, Ex234, Ex235, Ex236, Ex237 respectively;
Adopt following basic recipe: Cytochrome C (i.e. cytochrome C) 7.5mg, glucose 7.5mg, sodium sulfite 1.25mg, sodium sulfite 1.25mg, water for injection 0.35ml, and add Cys 0mg, 0.5mg, 1.0mg, 1.5mg, 2mg, 3mg, 5mg wherein, preparation method is with reference to embodiment 4, obtain 7 injectable powder formula, its numbering is designated as Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, Ex247 respectively.
test 3: the pharmaceutical property investigating each compositions sample
Sample: each sample Ex4 of embodiment 4 ~ embodiment 6, Ex5, Ex6, the sample Ex201 of additional embodiment 2, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex209, Ex210, sample Ex211, Ex212, Ex213, Ex214, Ex215, sample Ex216, Ex217, Ex218, Ex219, Ex220, sample Ex221, Ex222, Ex223, Ex224, Ex225, Ex226, Ex227, sample Ex231, Ex232, Ex233, Ex234, Ex235, Ex236, Ex237, sample Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, Ex247.
Method: each injectable powder sample is placed in 35 DEG C of dark places with reference to the method in test 1 and places 6 months (referred to as high-temperature treatment or 35 DEG C-June or can dispose the 35C-6 month in the present invention), concurrently each injectable powder sample is placed in 5 DEG C of dark places to place 6 months (can dispose referred to as low temperature in the present invention or 5 DEG C-June or 5 DEG C-June are disposed) simultaneously, check the particulate matter of each sample when June, add up each sample >=10 μm of particle numbers.Calculate the microgranule percent change of each sample.
Result: Ex201 in each sample of the embodiment 4 ~ embodiment 6 of testing and additional embodiment 2, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex211, Ex212, Ex213, Ex214, Ex216, Ex217, Ex218, Ex219, Ex221, Ex222, Ex223, Ex224, Ex225, Ex231, Ex232, Ex233, Ex234, Ex235, Ex241, Ex242, Ex243, Ex244, these cysteine consumptions of Ex245 are lower than the sample equaling 2mg (for every 7.5mg active component), their microgranule percent change (%) is all in 95 ~ 122% scopes, such as Ex4, Ex5, the microgranule percent change (%) of Ex6 three is respectively 107%, 112%, 98%, show very gratifying physical stability.
But for Ex209, Ex210, Ex215, Ex220, Ex226, Ex227, Ex236, Ex237, these cysteine consumptions of Ex246, Ex247 are higher than the sample of 2mg, and their microgranule percent change (%) is all in 177 ~ 293% scopes, the microgranule percent change (%) of such as both Ex209, Ex226 is respectively 241%, 207%, and display cannot gratifying physical stability.
Visible, for the physical stability characterized with microgranule percent change, add in injectable powder for the physical stability that product characterized with microgranule percent change lower than the cysteine equaling 2mg for 7.5mg active component be do not have influential.
test 4: investigate the active ingredient loss of each compositions sample in process for preparation
Sample: each sample Ex4 of embodiment 4 ~ embodiment 6, Ex5, Ex6, each sample Ex201 of additional embodiment 2, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex209, Ex210, Ex211, Ex212, Ex213, Ex214, Ex215, Ex216, Ex217, Ex218, Ex219, Ex220, Ex221, Ex222, Ex223, Ex224, Ex225, Ex226, Ex227, Ex231, Ex232, Ex233, Ex234, Ex235, Ex236, Ex237, Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, Ex247.
Method in reference test 2 is carried out, investigate the active ingredient loss (%) of each sample, result shows whole sample, and (they are all in the dosing process of preparation process (3), use 0.05 ~ 0.1% paper pulp) active ingredient loss (%) all in 0.23% ~ 3.04% scope, the active ingredient loss (%) of such as Ex4, Ex5, Ex6 three is respectively 1.27%, 0.93%, 1.12%.Visible, although embodiment 4 ~ embodiment 6 and additional embodiment 2 these use in preparation process (3) in lower than each sample of 0.1% paper pulp gained and be all added with cysteine, cysteine on dosing loss without impact.
test 5: the biological stability investigating each compositions sample
Sample: each sample Ex1 of embodiment 1 ~ embodiment 3, Ex2, Ex3, each sample Ex101 of additional embodiment 1, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex108, Ex109, Ex110, Ex111, Ex121, Ex122, Ex123, Ex124, Ex125, Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex137, Ex138, Ex139, Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex147, Ex148, Ex149, Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157, Ex161, Ex162, Ex163, Ex164, Ex165, Ex166, and each sample Ex4, Ex5, Ex6 of embodiment 4 ~ embodiment 6, each sample Ex201, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex209, Ex210 of additional embodiment 2, Ex211, Ex212, Ex213, Ex214, Ex215, Ex216, Ex217, Ex218, Ex219, Ex220, Ex221, Ex222, Ex223, Ex224, Ex225, Ex226, Ex227, Ex231, Ex232, Ex233, Ex234, Ex235, Ex236, Ex237, Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, Ex247, its numbering of three injectable powder samples prepared with reference to formula and the method for the embodiment 1 of CN103055305A, embodiment 2, embodiment 3 is respectively Co1, Co2, Co3.
Method: each injectable powder sample is placed in 35 DEG C of dark places and places 6 months (in the present invention can referred to as disposing in high-temperature treatment or 35 DEG C-June or 35 DEG C-June), concurrently each injectable powder sample is placed in 5 DEG C of dark places to place 6 months (can dispose referred to as low temperature in the present invention or 5 DEG C-June or 5 DEG C-June are disposed) simultaneously, measures the cytochrome C vigor of each sample when June.For each sample, calculate it relative to the relative activity (%) after K cryogenic treatment after high-temperature process, relative activity calculating formula is as follows:
Relative activity (%)=(vigor after the vigor ÷ low temperature disposal after high-temperature treatment) × 100%
Close to 100%, above-mentioned relative activity (%) more shows that the biological stability of product is better, less then product biological stability is poorer.
Result: for whole samples not adding Cys of embodiment 1 ~ embodiment 3 and additional embodiment 1, and for Ex201 in additional embodiment 2, Ex202, Ex203, Ex204, Ex211, Ex212, Ex213, Ex214, Ex215, Ex216, Ex217, Ex218, Ex219, Ex220, Ex221, Ex222, Ex231, Ex232 these add that Cys amounts are lower or what add is the sample of D-Cys or DL-cysteine, and for Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, but these interpolation Cys excipient of Ex247 are not the samples using mannitol, and Co1, Co2, the sample that these art methods of Co3 prepare, they through June process after relative activity (%) all lower than 79%, all in 57 ~ 79% scopes, such as Ex202, Ex244, Co1 three is respectively 74.4% at relative activity (%) after process in June, 62.5%, 76.2% (CN103055305A description table 1 contained embodiment 1 sample was through the vigor data of 6 months, a comparability is not had with the result at this place of the present invention relative activity (%), because the present invention disposes 35 DEG C-June, and CN103055305A table 1 data clearly do not record disposal temperature, and the computational methods of the two are also different),
But for embodiment 4 ~ embodiment 6 sample Ex4, Ex5, Ex6, the sample Ex205 of additional embodiment 2, Ex206, Ex207, Ex208, Ex209, Ex210, sample Ex223, Ex224, Ex225, Ex226, Ex227, sample Ex233, Ex234, Ex235, Ex236, Ex237 these with the addition of the sample of the Cys being more than or equal to 1mg/ml, they through June process after relative activity (%) all in 96 ~ 105% scopes, such as Ex4, Ex5, Ex6 three is respectively 99.6% at relative activity (%) after process in June, 102.7%, 101.2%.
The result of above test 5 shows, the cytochrome C compositions being added with more than 1mg/ml Cys in the present invention has good biological stability.
And the result testing 3 above shows, adding for 7.5mg cytochrome C amount in injectable powder for the physical stability that characterizes with microgranule percent change for product lower than the cysteine equaling 2mg is do not have influential, but then has adverse influence to the physical stability of product when interpolation is greater than the Cys of relative quantity 2mg.Visible, for taking into account physical stability and the biological stability of compositions, in the present composition, use the Cys of 1 ~ 2mg for 7.5mg cytochrome C amount to be useful especially.
test 6: the chemical stability investigating each compositions sample
Sample: each sample Ex1 of embodiment 1 ~ embodiment 3, Ex2, Ex3, each sample Ex101 of additional embodiment 1, Ex102, Ex103, Ex104, Ex105, Ex106, Ex107, Ex108, Ex109, Ex110, Ex111, Ex121, Ex122, Ex123, Ex124, Ex125, Ex131, Ex132, Ex133, Ex134, Ex135, Ex136, Ex137, Ex138, Ex139, Ex141, Ex142, Ex143, Ex144, Ex145, Ex146, Ex147, Ex148, Ex149, Ex151, Ex152, Ex153, Ex154, Ex155, Ex156, Ex157, Ex161, Ex162, Ex163, Ex164, Ex165, Ex166, and each sample Ex4, Ex5, Ex6 of embodiment 4 ~ embodiment 6, each sample Ex201, Ex202, Ex203, Ex204, Ex205, Ex206, Ex207, Ex208, Ex209, Ex210 of additional embodiment 2, Ex211, Ex212, Ex213, Ex214, Ex215, Ex216, Ex217, Ex218, Ex219, Ex220, Ex221, Ex222, Ex223, Ex224, Ex225, Ex226, Ex227, Ex231, Ex232, Ex233, Ex234, Ex235, Ex236, Ex237, Ex241, Ex242, Ex243, Ex244, Ex245, Ex246, Ex247.
Method: each injectable powder sample is placed in 35 DEG C of dark places and places 6 months (in the present invention can referred to as disposing in high-temperature treatment or 35 DEG C-June or 35 DEG C-June), concurrently each injectable powder sample is placed in 5 DEG C of dark places to place 6 months (can dispose referred to as low temperature in the present invention or 5 DEG C-June or 5 DEG C-June are disposed) simultaneously, measures the content (mg/ml) of cytochrome C in each sample when June.For each sample, calculate its active component content percent change (%) after high-temperature process, calculating formula is as follows:
Close to 0, above-mentioned active component content percent change (%) more shows that the chemical stability of product is better, larger then product chemical stability is poorer.
Result: the whole samples tested, after process in 6 months, active component content percent change (%), all in-0.53% to 4.36% scope, shows whole sample and all has good chemical stability.
Although many samples show good chemical stability, but they but demonstrate different biological stability features and demonstrate the physical stability characteristics characterized with microgranule situation of change, this species diversity completely cannot from formula each adjuvant function construable logical.
industrial applicability
The invention provides a kind of pharmaceutical composition such as injectable powder of cytochrome C.Cytochrome C pharmaceutical composition of the present invention can be used for the auxiliary treatment of various histanoxia first aid, the treatment of the myocardial ischemia that the dyspnea caused as carbon monoxide poisoning, hypnotics poisoning, cyanide poisoning, asphyxia of newborn, serious shock phase anoxia, cerebrovas-cularaccident, cerebral concussion sequela, anesthesia and pulmonary disease and various cardiac disorder cause.
Claims (16)
1. a pharmaceutical composition, it is aseptic freeze-dried injectable powder, wherein comprises: the cytochrome C of 7.5 weight portions, the freeze-dried excipient mannitol of 7.5 ~ 150 weight portions, the antioxidant of 1 ~ 5 weight portion, the Cys of 1 ~ 2 weight portion and optional acid-base modifier; Described antioxidant is selected from sodium sulfite, sodium sulfite and combination thereof; This lyophilized injectable powder is when the preparation of medicinal liquid through activated carbon adsorption processing procedure, and this activated carbon adsorption processing procedure is: in the medicinal liquid comprising active component, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, stir 10 ~ 30 minutes; The amount of the described paper pulp added together with active carbon, in dry filter paper, the concentration in medicinal liquid is 0.05% ~ 0.1% (w/v).
2. pharmaceutical composition according to claim 1, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of freeze-dried excipient is 7.5 ~ 100 weight portions.
3. pharmaceutical composition according to claim 1, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of freeze-dried excipient is 7.5 ~ 75 weight portions.
4. pharmaceutical composition according to claim 1, in the cytochrome C of every 7.5 weight portions in the material wherein comprised, the amount of antioxidant is 2 ~ 3 weight portions.
5. pharmaceutical composition according to claim 1, described acid-base modifier is selected from sodium hydroxide, potassium hydroxide, sodium dihydrogen phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, hydrochloric acid, phosphoric acid, nitric acid, sulphuric acid or its combination.
6. pharmaceutical composition according to claim 1, it is dissolved in water and/or dilutes the solution made containing 3mg cytochrome C in every 1ml, and the pH value of this solution is 6.0 ~ 7.5.
7. pharmaceutical composition according to claim 1, it is dissolved in water and/or dilutes the solution made containing 3mg cytochrome C in every 1ml, and the pH value of this solution is 6.5 ~ 7.5.
8. pharmaceutical composition according to claim 1, it is dissolved in water and/or dilutes the solution made containing 3mg cytochrome C in every 1ml, and the pH value of this solution is 7.0 ~ 7.5.
9. pharmaceutical composition according to claim 1, its solution before lyophilization also comprises water for injection except comprising cytochrome C, freeze-dried excipient, antioxidant, Cys and optional acid-base modifier, and the solid content of described solution is 1.5 ~ 20% (w/v).
10. pharmaceutical composition according to claim 9, the solid content of described solution is 2 ~ 15% (w/v).
11. pharmaceutical compositions according to claim 9, the solid content of described solution is 2 ~ 12% (w/v).
12. pharmaceutical compositions according to claim 9, it redissolves to substantially identical with solution before lyophilization volume with water for injection, and the solid content in gained solution is 1.5 ~ 20% (w/v).
13. pharmaceutical compositions according to claim 12, the solid content in gained solution is 2 ~ 15% (w/v).
14. pharmaceutical compositions according to claim 12, the solid content in gained solution is 2 ~ 12% (w/v).
15. according to the pharmaceutical composition of any one of claim 1-14, and it is prepared in accordance with the following methods and obtains:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection;
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50-60 DEG C, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, insulated and stirred 10 ~ 30 minutes;
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, if desired by the scope that acid-base modifier regulates this solution to conform with the regulations to pH;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.
The method of the pharmaceutical composition of 16. any one of preparation claim 1-15, the method comprises the following steps:
(1) each solid adjuvant material and cytochrome C are dissolved in appropriate water for injection;
(2) pH value to 7.0 ~ 7.2 of solution are regulated with acid-base modifier;
(3) be heated to 50-60 DEG C, add 0.05 ~ 0.5% (w/v) active carbon, and add paper pulp, insulated and stirred 10 ~ 30 minutes; The amount of the described paper pulp added together with active carbon, in dry filter paper, the concentration in medicinal liquid is 0.05% ~ 0.1% (w/v);
(4) step (3) gained medicinal liquid is first used filter paper filtering, then filter with the microporous filter membrane of 0.8 μm and 0.45 μm successively, to remove active carbon and paper pulp;
(5) benefit injects water to appropriate, stirs, and measures solution ph and optional mensuration active component content, if desired by the scope that acid-base modifier regulates this solution to conform with the regulations to pH;
(6) be filled with nitrogen, 4-6 DEG C of cold preservation 48h, use the microporous filter membrane aseptic filtration of 0.45 μm, 0.22 μm successively, filtrate is filled in cillin bottle, and lyophilization removing moisture, tamponade, to obtain final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310529187.XA CN103520123B (en) | 2013-10-31 | 2013-10-31 | Cytochrome C freeze-dried powder injection solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310529187.XA CN103520123B (en) | 2013-10-31 | 2013-10-31 | Cytochrome C freeze-dried powder injection solution |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103520123A CN103520123A (en) | 2014-01-22 |
| CN103520123B true CN103520123B (en) | 2015-03-04 |
Family
ID=49922714
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310529187.XA Active CN103520123B (en) | 2013-10-31 | 2013-10-31 | Cytochrome C freeze-dried powder injection solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103520123B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106986929A (en) * | 2017-04-19 | 2017-07-28 | 烟台东诚药业集团股份有限公司 | A kind of cromoci preparation method |
| CN110507815B (en) * | 2019-09-30 | 2023-02-14 | 成都通德药业有限公司 | Cytochrome C for injection and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103055305B (en) * | 2012-12-27 | 2014-06-18 | 马鞍山丰原制药有限公司 | A lyophilized preparation of a cytochrome C-containing pharmaceutical composition for injection and a preparation method thereof |
-
2013
- 2013-10-31 CN CN201310529187.XA patent/CN103520123B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN103520123A (en) | 2014-01-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11369587B2 (en) | Injectable pharmaceutical composition of tecovirimat and preparation method thereof | |
| US20110312914A1 (en) | Carbon monoxide removal agent | |
| JP2012233007A (en) | Octanoate-reduced human albumin | |
| KR20150022854A (en) | Manufacture of degarelix | |
| CN103536910B (en) | Cytochrome c injection | |
| CN103520123B (en) | Cytochrome C freeze-dried powder injection solution | |
| CN103251565A (en) | Voriconazole freeze-dried powder injection for injection and preparation method thereof | |
| EP3485886B1 (en) | Oral pharmaceutical composition of tecovirimat and preparation method therefor | |
| CN103222963B (en) | Somatostatin freeze-dried powder injection | |
| CN102614492B (en) | Liquid pharmaceutical composition containing echinocandin antifungal agent micafungin | |
| Cong et al. | Discovery of an orally effective double-stapled peptide for reducing ovariectomy-induced bone loss in mice | |
| CN103169674B (en) | Omeprazole sodium freeze-dried powder injection for injection | |
| CN104414977A (en) | Artesunate and L-arginine composition for injection and preparation method thereof | |
| US8911790B2 (en) | Solid composition containing a hypothiocyanite salt | |
| CN104706655B (en) | Meglumine cyclic adenosine for injecta powder-injection pharmaceutical composition and preparation method | |
| CN108606955A (en) | The Pharmaceutical composition and preparation method thereof of trainingization Xihai sea horse peptide | |
| CN104434803A (en) | Artesunate and L-lysine composition for injection and preparation method therof | |
| CN104069063B (en) | Fasudic hydrochloride pharmaceutical composition and preparation method thereof | |
| CN103055305A (en) | A lyophilized preparation of a cytochrome C-containing pharmaceutical composition for injection and a preparation method thereof | |
| CN103340831B (en) | Ganciclovir powder-injection for injection | |
| CN104721153B (en) | Injection aminomethylbenzoic acid freeze-drying powder-injection pharmaceutical composition | |
| CN103536567B (en) | Thymopentin (TP-5) powder injection | |
| CN107019675A (en) | Adenosine cyclophosphate for injection freeze drying powder injection pharmaceutical composition and quality control method and preparation method | |
| EP3274000B1 (en) | Dye molecule and dye preparations, in particular for use in surgical methods of ophthalmic surgery and for dyeing proteins | |
| CN103536898A (en) | Thymopentin (TP-5) drug composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 611531 Tiantaishan Pharmaceutical Co., Ltd., 88 Tianxing Avenue, Qionglai City, Chengdu City, Sichuan Province Patentee after: Chengdu Tiantaishan Pharmaceutical Co.,Ltd. Address before: 611531 Tiantaishan Pharmaceutical Co., Ltd., 88 Tianxing Avenue, Qionglai City, Chengdu City, Sichuan Province Patentee before: CHENGDU TIANTAISHAN PHARMACEUTICAL Co.,Ltd. |