CN103110663A - Extract and preparation - Google Patents
Extract and preparation Download PDFInfo
- Publication number
- CN103110663A CN103110663A CN2012105286260A CN201210528626A CN103110663A CN 103110663 A CN103110663 A CN 103110663A CN 2012105286260 A CN2012105286260 A CN 2012105286260A CN 201210528626 A CN201210528626 A CN 201210528626A CN 103110663 A CN103110663 A CN 103110663A
- Authority
- CN
- China
- Prior art keywords
- extract
- preparation
- vaccinia virus
- acetylneuraminic acid
- injection
- 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.)
- Granted
Links
Abstract
The invention aims at providing a vaccinia virus inoculated rabbit inflammatory skin extract and a preparation containing the extract as an effective component. An amount of N-acetyl neuraminic contained in the vaccinia virus inoculated rabbit inflammatory skin extract and the preparation containing the extract as an effective component is used as an indicator, and thus the quality of the extract and the preparation is ensured to be relatively stable. According to the method, the extract and the preparation with relatively stable quality has relatively strict guarantee on effectiveness and safety, thereby being quite useful.
Description
Technical field
The present invention relates to contain the vaccinia virus inoculation rabbit inflammation skin extract of N-acetylneuraminic acid of ormal weight and the preparation take this extract as effective ingredient etc.
Background technology
Pharmaceuticals are only to guarantee that its quality just can be subject to making the commodity of the license of sale.In Japan, in the 14th of medicine thing method, this is stipulated.In other various countries, at pharmaceuticals in nature, such processing is also same basically.Like this in pharmaceuticals quality guarantee that being considered to very important is because quality can guarantee effectiveness and the safety of pharmaceuticals.Quality does not obtain the pharmaceuticals guaranteed conversely speaking,, can not guarantee effectiveness and safety, just lacks the qualification as pharmaceuticals.
In Japan, the raw material that uses in the production as pharmaceuticals and the material that becomes the effective ingredient of pharmaceuticals are called as " former medicine ".In addition, former medicine and pharmaceuticals must be guaranteed its quality too.Reason is the quality that the quality of pharmaceuticals depends on former medicine.In addition, in the relevant decree of medicine thing of Japan, former medicine is defined as and is specifically designed to the pharmaceuticals of making other pharmaceuticals, and in definition, former medicine is comprised in pharmaceuticals.But, in this application, for convenient, sometimes dividing another name pharmaceuticals and former medicine, the pharmaceuticals of this moment refer to remove former medicine pharmaceuticals in addition.
Generally speaking, pharmaceuticals and former medicine are by the manufacture method manufacturing of regulation, thus pharmaceuticals and the former medicine of manufacturing specified quality.In addition, whether have in practice the quality of regulation, confirmed by the test of carrying out after making and inspection.When the result of such test and inspection does not meet predetermined benchmark, the not licensed listing of these pharmaceuticals and former medicine, sale etc.Operation is managed its quality to pharmaceuticals and former pencil like this.
Vaccinia virus inoculation rabbit inflammation skin extract (below be sometimes referred to as " this extract ") is to contain the extract that extracts the non-albuminous active substance that separates the inflammation skin histology of the rabbit that sends out pox from the inoculation vaccinia virus.Be liquid under the state of this extract after being extracted, but also can make solid by drying.
Contain the preparation that this extract is effective ingredient (below be sometimes referred to as " this preparation "), as described later, very useful as pharmaceuticals.At this moment, because this extract is the effective ingredient of this preparation, so this extract just becomes the former medicine of this preparation.As this preparation, the applicant makes in the concrete commodity of selling in Japan to be had " preparation that contains vaccinia virus inoculation rabbit inflammation skin extraction liquid " (trade name: NEUROTROPIN ［ registered trade mark ］ is hereinafter referred to as " NTP preparation ").In the NTP preparation, injection (hereinafter referred to as " NTP injection ") and tablet (hereinafter referred to as " NTP tablet ") are arranged, be medical pharmaceuticals (ethical drug).Vaccinia virus inoculation rabbit inflammation skin extraction liquid (hereinafter referred to as " NTP extracting solution ") as NTP preparation effective ingredient is the former medicine of NTP preparation.The NTP extracting solution is included in this extract, and NTP preparation (NTP injection and NTP tablet) is included in this preparation.
The indication of NTP injection is " Lumbago, neck shoulder wrist syndrome, symptomatic neuralgia, follow the creeping chill-abnormal perception of the scratching where it itches of dermatosis (eczema, dermatitis, urticaria), allergic rhinitis, SMON (SMON) sequela-pain ".The indication of NTP tablet is " postherpetic neuralgia, Lumbago, neck shoulder wrist syndrome, scapulohumeral periarthritis, osteoarthrisis deformans knee ".The NTP preparation is the preparation that the applicant formulates and exploitation obtains as pharmaceuticals.With regard to the NTP preparation, the excellent characteristics in its effectiveness and safety is subject to higher rating, is sold all the year round, has established firm status in the pharmaceuticals market of Japan.In addition, the NTP preparation is exported to China now, sells with the trade name of " Neurotropin/NEUROTROPIN ".Indication at the NTP of China preparation is identical with Japan.
Like this, this preparation is very useful as pharmaceuticals, and this extract is also very useful as the former medicine of this preparation.But as mentioned above, this extract is to extract the extract that separates from the inoculation vaccinia virus the inflammation skin histology of the rabbit of a pox.Therefore, this extract contains extremely many materials (composition), uses this preparation of this extract manufacturing also still to contain extremely many materials (composition).The quality of therefore, how to manage this extract and this preparation is very important item.
Many pharmaceuticals are that usually, these materials are by the compound of chemosynthesis with a kind or maximum 2～3 kinds of materials (composition) preparation as effective ingredient.Therefore, measure the content of this compound in said preparation, if contain the amount of regulation, the quality from the aspect of the active constituent content of said preparation is just guaranteed.But this extract is from the inoculation vaccinia virus and the extract of the inflammation skin histology of the rabbit of a pox, contains the material of utmost point multiple types.Take this extract this preparation as effective ingredient, certainly contain too the material of utmost point multiple types.Like this, this extract and this preparation are not as effective ingredient, so can not carry out effective ingredient is carried out as material such qualitative control in the common pharmaceuticals of special provision with specific a kind or number of substances.Therefore, this extract that the applicant makes and this preparation, i.e. the effective ingredient of NTP extracting solution and NTP preparation quantitatively (quantitative determination) is undertaken by the method for measuring its biological activity (tire, titer).
Said method is to use the biological test method (" day pharmacology will ", the 72nd volume, No. 5,573-584 page, 1976) of stress (cold load repeatedly) animal obtaining the analgesia coefficient as the SART of the pain threshold chronic stress animal lower than the intact animal.The method of putting down in writing by the document, using stress (cold load repeatedly) animal as the SART of the pain threshold chronic stress animal lower than the intact animal, carries out the analgesic test by inflammation foot pressurization (Randall-Selitto) method and obtains the analgesia coefficient.The method is in the afterbody of mice the stimulation of exerting pressure, as index, to measure the method for analgesic effect until mice shows the pressurization weight of escape reaction.So-called analgesia coefficient be will throw with medicine after the pressurization weight that measure throw divided by medicine the value that the pressurization weight with front mensuration obtains.About NTP extracting solution and NTP preparation, positive as analgesic effect with the situation of the analgesia coefficient more than the certain value that shows applicant's regulation, obtain the ratio that is judged as positive animal number of elements, as analgesia effective percentage (%).From this value, obtain ED according to the result of measuring for the standard substance of various concentration for dilution
50Value." Neurotropin unit (NU) " of the unit of the biological activity (tiring) that uses about the NTP preparation as the applicant is by with ED
50Value is for throwing the Mouse Weight with amount 100mg/kg() the activity that shows of this extract 1mg be made as 1Neurotropin unit and define.Then, measure ED about each NTP preparation
50Value compares quantitative analgesic activities (effective ingredient amount) with standard substance.Below, in this application, the yardstick as the effective ingredient amount (tiring) of this extract and this preparation uses " unit " such expression, but in fact with NTP extracting solution and NTP preparation in " Neurotropin unit " same meaning of using.
But, this extract and this preparation of making the applicant, namely in NTP extracting solution and NTP preparation, above-mentioned analgesic activities quantitatively beyond, also carry out following a plurality of validation tests, regulation must meet these tests.
The amino acid whose validation test of being undertaken by liquid chromatography
The confirmation test of the ultraviolet section absorbing material that is undertaken by the UV, visible light determination of absorbance
The confirmation test of the phosphorus that is undertaken by the chromogenic reaction method
The confirmation test of the nucleic acid base that is undertaken by liquid chromatography
The kallidinogenase sample material that is undertaken by in vitro test method(s) produces inhibiting validation test
Generally speaking, for making from the extract of the such biology of animal and plant and containing the maker that this extract is the preparation of effective ingredient, due to the time, work, the expense that do not need for test and inspection, the manufacture underproof probability that becomes also reduces in addition, so the benchmark that said preparation must meet is favourable less.But the viewpoint of collateral security extract as described above and preparation quality is set out, and the benchmark that hope should meet is more strictly stipulated.The present invention is from such a viewpoint, about this extract and this preparation that the applicant makes, furthers investigate suitable benchmark, the invention that found that as the new benchmark that should meet.Thus, the quality of this extract and this preparation becomes more stable.In addition, thus, effectiveness and the safety of this extract and this preparation are subject to more strictly guaranteeing.
As the document that discloses this extract or this preparation, there is patent documentation 1 to 3.In these documents relevant for the record of the content of the aminoacid in this extract or this preparation and nucleic acid base.In addition, identical about this extract or this preparation, in patent documentation 4 relevant for the record of silicon class content.But, in these patent documentations 1 to 4, contain the such predetermined substance of how many N-acetylneuraminic acids but without any record about this extract or this preparation.Moreover, in these documents, about the N-acetylneuraminic acid content in this extract or this preparation had not both been put down in writing not enlightenment as the index that is used for this extract or the qualitative control of this preparation yet.
The prior art document
Patent documentation
Patent documentation 1: the open CN1205233A communique of Chinese patent
Patent documentation 2: international open WO2004/060381 communique
Patent documentation 3: the open CN1613305A communique of Chinese patent
Patent documentation 4: Japanese kokai publication hei 7-97336 communique
Summary of the invention
Invent problem to be solved
As mentioned above, in this extract and this preparation, do not identify that one matter is as effective ingredient.Therefore, the quality of this extract and this preparation, the bioactivity that is undertaken by the biological test of stress mice with SART obtaining the coefficient that eases pain and a plurality of validation tests etc. are guaranteed.In addition, however, the various compositions that comprise in this extract and this preparation inevitably have the deviation of certain degree in each is made batch.But, since this preparation uses as the pharmaceuticals for the treatment of disease, just wish that its quality keeps certain as far as possible.More stably guaranteeing the quality of this extract and this preparation, be related to and make effectiveness and the safety of this extract and this preparation more constant, is very significant.
Be used for solving the method for problem
The present invention guarantees the invention of the quality of this extract and this preparation with the amount of the N-acetylneuraminic acid that contains as index in this extract and this preparation.Based on this, the invention provides: contain this extract of N-acetylneuraminic acid of defined amount and this preparation, by the content management of N-acetylneuraminic acid or estimate this extract and this preparation quality method or carry out with such method this extract and this preparation that qualitative control obtains.
The effect of invention
This extract that the present invention relates to and this preparation are extract and the preparations that contains the N-acetylneuraminic acid of ormal weight.Thus, the quality of this extract and this preparation obtains more stable guaranteeing, effectiveness and the safety of this extract and this preparation become more constant.
The specific embodiment
N-acetylneuraminic acid is a kind of of sialic acid.Sialic acid is the general name of the acyl derivative of neuraminic acid, has multiple sialic acid at occurring in nature, and wherein, N-acetylneuraminic acid is maximum, and the NeuGc ALPHA2-3Gal second is many.Sialic acid extensively distributes in vivo as the constituent of glycoprotein, glycolipid matter, glycopeptide etc., is present in especially the surface of cell membrane of animal or antibacterial, is bearing the important biological functions such as specific recognition mechanism that participate in cell.In addition, sialic acid comes into one's own in medical science and pharmacy as the material relevant to cancer, inflammation, immunity, viral infection, cell differentiation, hormone receptor etc., is carrying out various researchs about the biological component, sialic acid and the derivant thereof that contain sialic acid.
About this sialic acid, especially in regard to N-acetylneuraminic acid; the applicant is surprised to find that this material has analgesic activity; have especially the effect to the non-inflammatory pain disease (non-inflammatory pain disease) of neuropathic pain (neuropathic pain disease) etc., obtained patent (Japan's special permission No. 4005115, Chinese Granted publication CN101600438B) based on such discovery.Because it is also therapeutic effect to neuropathic pain etc. that the applicant makes the feature of the NTP preparation of sale, so as the index of the quality that is used for more strictly managing this extract and this preparation, N-acetylneuraminic acid is rational material (composition).
Then, the method for making this extract and this preparation is described.
This extract can be sent out by inoculation vaccinia virus in the skin that is captured in rabbit the inflammation skin of pox, carry out fragmentation, after adding the extraction solvent processing, remove tissue, removing albumen processes, make it be adsorbed in adsorbent under acid condition it, then under alkali condition, the effective ingredient eluting is obtained.
Wherein, vaccinia virus (vaccinia virus) can be the vaccinia virus of any strain.As an example, can enumerate Li Site (Lister) strain, Dalian (Dairen) strain, Ikeda (Ikeda) strain, EM-63 strain, New York community health office (New York City Board of Health) strain etc.
In addition, as long as belonging to Lagomorpha, rabbit just can be rabbit arbitrarily.As an example, rabbit, rabbit (with the rabbit of rabbit domestication), hare (Japanese hare), pika, Lepus timidus etc. are arranged.Wherein, be fit to use rabbit.In Japan, have from the past raised as domestic animal or animal for research and the frequent rabbit that is called domestic rabbit (イ エ ウ サ ギ) of using, but this is also the another name of rabbit.There are a plurality of kinds (breed) in rabbit, but can preferably use the kind that is called Japan's white kind and New Zealand's white kind (New Zealand is white).
As the basic abstraction process of this extract, for example, use following such operation.
(A) gather the intradermal vaccination vaccinia virus, make the skin histology of the rabbit of its pox, pox organization is broken, after the extraction solvent that adds entry, phenol water, normal saline or be added with the glycerol liquor etc. of phenol carries out the extraction process of a few days, by filtering or centrifugalize, obtain the crude extract (filtrate or supernatant) that tissue is removed.
The crude extract that (B) will obtain in (A) is adjusted into acid pH and heats, and filtration or centrifugalize remove albumen and process.Then, the pH that will be adjusted into except the solution after albumen alkalescence heats, and refilters or centrifugalize, obtains except filtrate or supernatant after the albumen processing.
The filtrate that (C) will obtain in (B) or supernatant are adjusted into acid pH, make it be adsorbed in the adsorbent of activated carbon, Kaolin etc.
(D) the extraction solvent that adds entry etc. in (C) in the adsorbent that obtains is adjusted into the pH of alkalescence, by with the adsorption component eluting, obtains vaccinia virus inoculation rabbit inflammation skin extract (this extract).
Be more than basic operation, followingly will narrate in more detail these operations.
About (A)
Be captured in the skin of rabbit vaccinia virus is carried out the inflammation skin histology that intradermal vaccination makes its pox.The skin histology that gathers cleans, sterilizes with phenol solution etc.Broken this inflammation skin histology adds the extraction solvent of its 1 to 5 times of amount.Wherein, so-called fragmentation refers to use meat grinder etc. to be broken for meticulously the minced meat shape.In addition, as extracting solvent, can use distilled water, normal saline, faintly acid to weakly alkaline buffer etc., also can suitably add the salt etc. of the stabilizing agent, sodium chloride, potassium chloride, magnesium chloride etc. of the sterilization-antiseptic, glycerol etc. of phenol etc.At this moment, also can destroy by the processing of freezing thawing, ultrasound wave, cell membrane lyase or surfactant etc. cell tissue makes extraction become easy.The suspension that obtains was placed 5 to 12.During this, can the limit agitation as appropriate or do not stir the limit and be warmed to 30 to 45 ℃.Remove tissue by the liquid filtering that will obtain or centrifugalize etc., obtain crude extract (filtrate or supernatant).
About (B)
The crude extract that obtains in (A) is removed albumen to be processed.Except albumen can be implemented by the known method that usually carries out, can application of heat process, by processing, the isoelectric precipitation of protein denaturant (such as acid, alkali, urea, guanidine, acetone and other organic solvent etc.), the method for saltouing etc.Then, by removing the usual method of insoluble matter, such as the filtration of using filter paper (cellulose, celluloid etc.), glass filter, diatomite product, plug thatch screen plate etc., ultrafiltration, centrifugalize etc., filtrate or the supernatant of the insoluble protein of separating out of being removed.
About (C)
The filtrate that will obtain in (B) or supernatant are adjusted into acidity, preferably are adjusted into pH3.5 to 5.5, carry out the adsorption operations to adsorbent.As the adsorbent that can use, can enumerate activated carbon, Kaolin etc., add adsorbent and stir or make extracting solution pass through the adsorbent packed column in extracting solution, can make effective ingredient be adsorbed in this adsorbent.When adding adsorbent in extracting solution, remove solution by filter or centrifugalize etc., can access the adsorbent that absorption has active component.
About (D)
In order to make eluting (disengaging) in the adsorbent that active component obtains from (C), add eluting solvent in this adsorbent, be adjusted into alkalescence, preferably be adjusted into pH9 to 12, in room temperature or suitably heating or stir and carry out eluting, to filter or the usual way such as centrifugalize is removed adsorbent.As the eluting solvent that uses, can use basic solvent, for example, can use the water that is adjusted into alkaline pH, methanol, ethanol, isopropyl alcohol etc. or their suitable mixed solution, can preferably use the water that is adjusted into pH9 to 12.The amount of eluting solvent can suitably be set.Use as former medicine in order to operate like this eluent that obtains, pH suitably can be adjusted near neutral etc., finally obtain vaccinia virus inoculation rabbit inflammation skin extract (this extract).
This extract is liquid when making, therefore also can be by the extract that suitably concentrates, desired concentration is made in dilution.When making preparation from this extract, preferably apply heat sterilization and process.In order to make injection, for example, the solution that can add the preparations such as sodium chloride and normal saline etc. to ooze.In addition, also can carry out by this extract to liquid condition the operation of suitable concentrate drying curing etc., make the solid preparation for oral administration of tablet etc.From the concrete grammar of the such solid preparation for oral administration of this extract manufacturing, on the books in the description of No. the 4883798th, No. the 3818657th, Japan special permission and Japan's special permission.This preparation is the injection that obtains of like this operation or solid preparation for oral administration etc.
The content of the N-acetylneuraminic acid in this extract and this preparation can be measured by the common quantitative method of using.Particularly, for example, can use the algoscopy of utilizing liquid chromatography mass analyser (LC-MS) or capillary electrophoresis interfaced with mass spectrometry instrument (CE-MS), by DMB(1,2-diaminourea-4,5-methylenedioxy benzene) fluorescence labelling method, enzyme process etc.In any a kind of method, can both use to proofread and correct the calibration trace of the N-acetylneuraminic acid sample production of use, the N-acetylneuraminic acid in this extract and this preparation is carried out quantitatively.
Measure the content of N-acetylneuraminic acid contained in this extract that the applicant makes and this preparation with method as described above.Its result, in this extract and this preparation, although deviation is arranged, every 1 unit contains the above N-acetylneuraminic acid of 4000ng.Furthermore, in this extract and this preparation, every 1 unit contains the N-acetylneuraminic acid of 4000～18000ng.Therefore, with the content of the N-acetylneuraminic acid in this extract and this preparation during as the quality of this extract of INDEX MANAGEMENT and this preparation, judgement can be set as benchmark with such content.In addition, for the applicant, when having confirmed on the manufacture method of this extract and this preparation, change to be arranged, the content of N-acetylneuraminic acid is sometimes lower than below every 1 4000ng of unit.
Wherein, as seen from the above description, so-called " every 1 unit " refers to the meaning of the every active constituent content in this extract and this preparation.In this extract (NTP extracting solution) that the applicant makes, contain the effective ingredient of 1.2 units/mL.This preparation (NTP injection) of using at the injection that uses its manufacturing contains the effective ingredient of 1.2 units/mL too.In the NTP injection, there are the injection of capacity 3mL and the injection of 1mL.Therefore, contain the effective ingredient of 3.6 units in the preparation of 3mL, contain the effective ingredient of 1.2 units in the preparation of 1mL.On the other hand, in for oral use the preparation (NTP tablet) that the applicant makes, every 1 effective ingredient that contains 4 units.
Yet this extract can concentrate or dilute.In addition, this preparation also can be made the preparation that contains various units.At this moment, the amount of contained effective ingredient also can change in every part of component of this extract and this preparation (every 1mL, every 1mg, every 1 pipe, every 1 etc.).Therefore, basically mean with the content of the relationship between quantities regulation N-acetylneuraminic acid of the effective ingredient of this extract and this preparation.Reason is that this relates to the cause with the relation of the effectiveness of this extract and this preparation and safety.Therefore, the applicant stipulates the content of the N-acetylneuraminic acid in this extract and this preparation with the content (" every 1 unit ") of every effective ingredient.On the other hand, the applicant sells the NTP preparation from present manufacturing, with every 1mL and every 1 pipe of concrete injection or stipulate with every 1 of tablet, also due to throwing and the relationship between quantities on have certain meaning, so also carry out such regulation.
But, in the NTP injection, in Japan and Chinese Counterfeit Item (generic drug) or the similar formulations (similardrug) (following these are called " other company's injection ") that has applicant's company (hereinafter referred to as " other company ") manufacturing in addition.These other company's injections also similarly use the SART stress mice with the NTP preparation, take analgesic effect as index, carry out as the vaccinia virus inoculation rabbit inflammation skin extract (this extract) of effective ingredient quantitatively.In expression, the active constituent content of these other company's injections, as as " ROSEMORGEN In. " (registered trade mark) of a plurality of other company's injections of Japan, " NABUTOPINInj. " (registered trade mark) and " NOLPORT Inj. " (registered trade mark), existing simple table is shown the injection of " unit ", also has conduct to be expressed as like that the injection of " Analgecine unit " or " AGC unit " in " grace is suitable/ANALGECINE again " (registered trade mark) of other company's injection of China.But, even in these any preparations, the content of effective ingredient is similarly also 1.2 units or 1.2Analgecine(1.2AGC for every 1mL with the NTP injection) unit, contain 3.6 units or 3.6Analgecine(3.6AGC in goods 1 pipe of 3mL dress) unit.As a result, just method for expressing is different in " unit " or " Analgecine unit " that " the Neurotropin unit " that uses in the NTP preparation and other company use, and is identical yardstick as the unit of regulation active constituent content.Therefore, in this application, NTP preparation not only also comprises the injection of other all companies, all uses " unit " as the expression that the expression effective ingredient is the content of this extract.Like this, a plurality of with the preparation of vaccinia virus inoculation rabbit inflammation skin extract as effective ingredient, with " unit " expression effective ingredient, with regard to said preparation, " unit " expression is clear and definite for those skilled in the art.
From as can be known above, in this application, " this preparation " is the concept that comprises NTP preparation (NTP injection and NTP tablet) and other company's injection.In addition, when other company will make, sell tablet (hereinafter referred to as " other company's tablet ") as Counterfeit Item or the similar product of NTP preparation, be also the concept that also comprises other company's tablet.
Below, represent this extract and the concrete example of this preparation manufacturing and the measurement result of the N-acetylneuraminic acid content in this extract and this preparation as embodiment, but the present invention is not subjected to any restriction of the record of these embodiment.
Embodiment
The manufacturing of this extract of embodiment 1()
The skin of sending out pox is downcut and gathered to intradermal vaccination vaccinia virus in the skin of healthy mature rabbit.After the skin that gathers cleans, sterilizes with phenol solution, remove unnecessary phenol solution, carry out fragmentation, add phenol solution to mix, place after 3～7 days, and then heat at 35～40 ℃ in stirring limit on the 3rd～4, limit.Then, adjust to pH4.5～5.2 with the extracting solution that hydrochloric acid obtains solid-liquid separation, after 30 minutes, remove by filter albumen with 90～100 ℃ of heat treated.With sodium hydroxide, filtrate is adjusted to pH9.0～9.5 again, after 15 minutes, carry out solid-liquid separation with 90～100 ℃ of heat treated.
Adjust to pH4.0～4.3 with what hydrochloric acid will obtain except protein liquid, add the activated carbon except 2% amount of protein liquid quality, stir after 2 hours, carry out solid-liquid separation.Add water in the activated carbon that gathers, adjust to pH9.5～10 with sodium hydroxide, after stirring 90～100 minutes with 60 ℃, carry out centrifugalize and obtain supernatant.Add entry again in the activated carbon with the centrifugalize precipitation after, adjust to pH10.5～11 with sodium hydroxide, after stirring 90～100 minutes with 60 ℃, carry out centrifugalize and obtain supernatant.Merge two supernatant, with the hydrochloric acid neutralization, obtain this extract.
The assay method of embodiment 2(N-acetyl neuraminic acid content)
The following N-acetylneuraminic acid content of measuring this extract and this preparation with high performance liquid chromatography mass spectrometer (LC-MS).
With this extract (1.2 units/mL), inject LC-MS of 10 times of dilutions of water according to embodiment 1 manufacturing.
Use this preparation (NTP injection) of making according to this extract of embodiment 1 manufacturing too with 10 times of dilutions of water, inject LC-MS.
Use this preparation (NTP tablet) of making according to this extract of embodiment 1 manufacturing, 3 are cleaned 3 times with methanol/chloroform (1: 1) 3mL, except the striping coatings, after drying, add 12mL water to hang turbid (1 unit/mL), after centrifugal with supernatant with 10 times of dilutions of water, inject LC-MS.
With regard to N-acetylneuraminic acid, the standard solution of obtained aqueous solution is made calibration trace.
1100 series that LC-MS uses Agilent to produce in HPLC section use API3000(Applied Biosystems/MDS Sciex to produce in mass spectrometer).Analysis condition is as follows.
Post: Inertsil ODS-3(Φ 2.1 * 150mm)
Column temperature: 25 ℃, flow velocity: 200 μ L/ minutes
Mobile phase: methanol/0.05% formic acid
Methanol %/minute: 0/0-0/1-44/8-100/8.1-100/11
Sample size: 5 μ L, sample room design temperature: 4 ℃
Detect: cation detects MRM
The condition determination of expression LC-MS in table 1.The meaning of each parameter in table 1 is as follows.
DP: the voltage that puts on orifice plate
FP: the voltage that puts on focusing ring
CE: collision energy
CXP: the voltage that puts on the Q2 outlet
NEB: the pressure of nebulizer gas
CUR: the pressure of gas curtain gas
IS: the voltage of ion spraying day with fog
CAD: the pressure of collision gas
TEM: impeller gas temperature (turbo gas temperature)
［ table 1 ］
| Detect ion (Q1/Q3) | DP | FP | CE | CXP | NEB | CUR | IS | CAD | TEM |
| 310/274 | 20 | 100 | 20 | 20 | 12 | 10 | 5000 | 8 | 450 |
The measurement result of the N-acetylneuraminic acid content of this extract of embodiment 3()
Expression measures with the method for record in above-described embodiment 2 result that obtains with the N-acetylneuraminic acid content in this extract in table 2.Active constituent content in this extract is 1.2 units/mL.With every 1 unit ("/unit ") of this extract and every 1mL("/mL " of this extract) both represent the content of the N-acetylneuraminic acid in this extract.In addition, the A in batch sequence number～C mark represents the difference of applicant's manufacturing site location (facility).In addition, measured value is unified to 3 of significant digits (below, identical in whole measured values).
［ table 2 ］
The measurement result 1 of the N-acetylneuraminic acid content of this preparation of embodiment 4()
In this preparation, expression measures with the method for record in above-described embodiment 2 result that obtains with the N-acetylneuraminic acid content in NTP injection (every 1mL contains 1.2 units) in table 3.Result is with every 1 unit ("/unit ") of this preparation effective ingredient, every 1mL("/mL " of this preparation) and each content of every 1 pipe of ampoul tube ("/pipe ") of 3mL dress represent.
［ table 3 ］
The measurement result 2 of the N-acetylneuraminic acid content of this preparation of embodiment 5()
In this preparation, expression measures with the method for record in above-described embodiment 2 result that obtains with the N-acetylneuraminic acid content in NTP tablet (every 1 contains 4 units) in table 4.Result represents with every 1 unit ("/unit ") of effective ingredient and each content of every 1 ("/sheet ").
［ table 4 ］
The measurement result of the N-acetylneuraminic acid content of other company's injection of comparative example 1()
Then, represent similarly to measure with the NTP injection result of the N-acetylneuraminic acid content in other company's injection in table 5.In addition, " ROSEMORGEN Inj. " be the application sell end when filing an application, on market non-existent batch, following measurement result is applicant's past to buy the result of measuring on market.
［ table 5 ］
From above result as can be known, even identical vaccinia virus inoculation rabbit inflammation skin extraction liquid formulation, according to manufacturing company, the content of N-acetylneuraminic acid also has a great difference (table 3～5).In addition, also also comprise as can be known the applicant interior, even the preparation of same companies also has significantly different situation (table 3～5) of N-acetylneuraminic acid content.About the vaccinia virus inoculation rabbit inflammation skin extraction liquid as the former medicine of vaccinia virus inoculation rabbit inflammation skin extraction liquid formulation, because can not obtain the product of other company, so can only measure the vaccinia virus inoculation rabbit inflammation skin extraction liquid (NTP extracting solution) that the applicant makes.And, for this also as can be known, deviation (table 2) is to a certain degree arranged in the content of N-acetylneuraminic acid.
But, in this extract and this preparation that the applicant makes, contain the above N-acetylneuraminic acid (table 2～table 4) of 4000ng/ unit.In addition, in this extract and this preparation that the applicant makes, contain N-acetylneuraminic acid with the scope of 4000～22500ng/ unit.On the other hand, in this preparation that other company makes, there is not the preparation (table 5) with high like this N-acetylneuraminic acid content.Although do not know which kind of reason is the difference of the N-acetylneuraminic acid content in this preparation that each company makes result from, strongly infer it is to result from the difference of manufacture method of this preparation of each company.In a word; due to as mentioned above; N-acetylneuraminic acid has analgesic activity; has especially the effect for non-inflammatory pain diseases such as neuropathic pains; so be interpreted as that as the applicant this material (composition) is many than other company's content in this preparation (NTP injection) that the applicant makes, itself become feature and be preferred feature.
As mentioned above, the N-acetylneuraminic acid that contains in this extract and this preparation more than 4000ng/ unit is the feature of this extract and this preparation.Similarly; in this extract and this preparation; when measuring with the assay method (hereinafter referred to as " 10 times of dilution methods ") of this extract of record in embodiment 2 and the N-acetylneuraminic acid content in this preparation (injection, tablet), contain N-acetylneuraminic acid as the feature of this extract and this preparation take the scope of 4000～15000ng/ unit.The dilution ratio that utilization will be obtained by water in 10 times of dilution methods is made as the operation of 10 times; be set as when waiting assay method (hereinafter referred to as " other dilution methods ") of other dilution ratio to measure N-acetylneuraminic acid content in this extract and this preparation with 50 times of water dilutions or 250 times of dilutions, contain N-acetylneuraminic acid as the feature of this extract and this preparation take the scope of 4000～22500ng/ unit.Wherein, during than 10 times of dilution methods, on be limited to the reason of 1.5 times and narrate in the back.
In addition, in this preparation, contain the injection (hereinafter referred to as " this injection ") of 1.2 units about every 1mL, to contain N-acetylneuraminic acid more than 4800ng/mL as the feature of this injection.Similarly, during with 10 times of By Dilutions, contain N-acetylneuraminic acid as the feature of this injection take the scope of 4000～18000ng/ unit.During with the content of the N-acetylneuraminic acid in other these injections of By Dilution, contain N-acetylneuraminic acid as the feature of this injection take the scope of 4800～27000ng/mL.In addition, when this injection is the injection of ampoul tube dress of 3mL capacity (following such injection is called " this 3mL fill injection "), the N-acetylneuraminic acid that every 1 pipe contains more than the 14400ng/ pipe is the feature that this 3mL fills injection.Similarly, during with 10 times of By Dilutions, contain N-acetylneuraminic acid as the feature of this 3mL dress injection take the scope of 14400～54000ng/ unit.During with the content of the N-acetylneuraminic acid in this 3mL of other By Dilutions dress injection, contain N-acetylneuraminic acid as the feature of this 3mL dress injection take the scope of 14400～81000ng/ pipe.Wherein, during than 10 times of dilution methods, on be limited to 1.5 times reason also in the back the narration.
In addition, in this preparation, about every 1 tablet tablet (hereinafter referred to as " this tablet ") that contains 4 units, to contain N-acetylneuraminic acid more than the 16000ng/ sheet as the feature of this tablet.Similarly, during with 10 times of By Dilutions, contain N-acetylneuraminic acid as the feature of this tablet take the scope of 16000～60000ng/ unit.During with the N-acetylneuraminic acid content in other these tablets of By Dilution, contain N-acetylneuraminic acid as the feature of this tablet take the scope of 16000～90000ng/ sheet.Wherein, during than 10 times of dilution methods, on be limited to 1.5 times reason also in the back the narration.
From above-mentioned situation, can take the N-acetylneuraminic acid content in this extract and this preparation as index, manage the quality of this extract and this preparation.That is, measure the content of the N-acetylneuraminic acid of this extract and this preparation, if it is more than 4000ng/ unit, it is qualified just can be evaluated as.In addition; also can be with the N-acetylneuraminic acid content in this extract and this preparation be the scope of 4000～22500ng/ unit during take 10 times of By Dilutions during as 4000～15000ng/ unit, take other By Dilutions, be made as the judgment standard of the qualification of this extract and this preparation.
During this injection, when the N-acetylneuraminic acid content in this injection is 4800ng/mL when above, it is qualified to be evaluated as.In addition, can be with the N-acetylneuraminic acid content in this injection during take 10 times of By Dilutions as 4800～18000ng/ unit, be made as the judgment standard of this injection qualification during take other By Dilutions as the scope of 4800～27000ng/mL.In addition; similarly when this 3mL dress injection, can be to be made as the judgment standard of this 3mL dress injection qualification during more than 14400ng/mL or take 10 times of By Dilutions during as 14400～54000ng/ unit, take other By Dilutions as the scope of 14400～81000ng/mL with the content of N-acetylneuraminic acid.
During this tablet, when the N-acetylneuraminic acid content in this tablet is 16000ng/ sheet when above, it is qualified to be evaluated as.In addition, can be with the N-acetylneuraminic acid content in this tablet be made as the judgment standard of this tablet qualification during take 10 times of By Dilutions during as 16000～60000ng/ unit, take other By Dilutions as the scope of 16000～90000ng/ sheet.
In addition; the numerical value that is equivalent to the upper limit during with the content of the above-mentioned N-acetylneuraminic acid of Range Representation of lower limit～upper limit; when being measured by other dilution process; become than the measurement result that obtains in embodiment 3～5 (table 2～4) much bigger (approximately 1.5 times of left and right), because following reason causes.
That is, in the above-described embodiments, the content of N-acetylneuraminic acid is measured by LC-MS, but N-acetylneuraminic acid easily is subject to being mingled with the impact (ion inhibition) of ion.Inventors of the present invention measure 10 times of the sample dilutions of this extract etc. basically, but dilution is when being 50 times or 250 times, the high value of measured value when determining than 10 times of dilutions.This phenomenon can be thought to descend because be mingled with the impact of ion concentration when dilution factor is high.The result that inventors of the present invention relatively verify with the quantitative values of each dilution ratio in a plurality of samples, when having confirmed that dilution is 50 times or 250 times, the value (table 6) of the maximum 1.5 times of left and right when determining 10 times of dilutions.Based on this fact, in this application, set as mentioned above the numerical value that is equivalent to the N-acetylneuraminic acid upper content limit.
［ table 6 ］
From above content, can derive following content as the present invention.But these are illustration, and the present invention is not limited by these.
(1) every 1 unit contains the vaccinia virus inoculation rabbit inflammation skin extract of the above N-acetylneuraminic acid of 4000ng.
(2) every 1 unit contains the vaccinia virus inoculation rabbit inflammation skin extract of the N-acetylneuraminic acid of 4000～22500ng.
(3) a kind of vaccinia virus inoculation rabbit inflammation skin extract as liquid, wherein, every 1mL contains the above N-acetylneuraminic acid of 4800ng.
(4) a kind of vaccinia virus inoculation rabbit inflammation skin extract as liquid, wherein, every 1mL contains the N-acetylneuraminic acid of 4800～27000ng.
(5) a kind of preparation that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 unit contains the above N-acetylneuraminic acid of 4000ng.
(6) a kind of preparation that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 unit contains the N-acetylneuraminic acid of 4000～22500ng.
(7) a kind of injection that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1mL contains the above N-acetylneuraminic acid of 4800ng.
(8) a kind of injection that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1mL contains the N-acetylneuraminic acid of 4800～27000ng.
(9) a kind of injection that contains the 3mL dress of vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 pipe contains the above N-acetylneuraminic acid of 14400ng.
(10) a kind of injection that contains the 3mL dress of vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 pipe contains the N-acetylneuraminic acid of 14400～81000ng.
(11) a kind of tablet that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 contains the above N-acetylneuraminic acid of 16000ng.
(12) a kind of tablet that contains vaccinia virus inoculation rabbit inflammation skin extract, wherein, every 1 N-acetylneuraminic acid that contains 16000～90000ng.
(13) as the preparation of putting down in writing in (5) or (6) of analgesics.
(14) as the injection of putting down in writing in any one in (7)～(10) of analgesics.
(15) as the tablet of putting down in writing in (11) or (12) of analgesics.
(16) manage the method for this extract quality by the amount of measuring the contained N-acetylneuraminic acid of vaccinia virus inoculation rabbit inflammation skin extract.
(17) method of the management vaccinia virus of record inoculation rabbit inflammation skin extract quality in (16); wherein; in every 1 unit of this extract, the amount of the N-acetylneuraminic acid that this extract is contained is 4000ng when above, and the quality of setting this extract is qualified.
(18) method of the management vaccinia virus of record inoculation rabbit inflammation skin extract quality in (16); wherein; in every 1 unit of this extract, when the amount of the N-acetylneuraminic acid that this extract is contained was 4000～22500ng, the quality of setting this extract was qualified.
(19) carry out with the method for putting down in writing in any one in (16)～(18) the vaccinia virus inoculation rabbit inflammation skin extract that qualitative control obtains.
(20) manage the method for said preparation quality by the amount that is determined at N-acetylneuraminic acid contained in the preparation that contains vaccinia virus inoculation rabbit inflammation skin extract.
(21) management of record contains the method that vaccinia virus is inoculated the preparation quality of rabbit inflammation skin extract in (20); wherein; in every 1 unit of vaccinia virus in said preparation inoculation rabbit inflammation skin extract; in said preparation, the amount of contained N-acetylneuraminic acid is 4000ng when above, and the quality of setting said preparation is qualified.
(22) management of record contains the method that vaccinia virus is inoculated the preparation quality of rabbit inflammation skin extract in (20); wherein; in every 1 unit of vaccinia virus in said preparation inoculation rabbit inflammation skin extract; when in said preparation, the amount of contained N-acetylneuraminic acid was 4000～22500ng, the quality of setting said preparation was qualified.
(23) carry out with the method for any one record in (20)～(22) preparation that qualitative control obtains.
(24) in the injection that contains vaccinia virus inoculation rabbit inflammation skin extract, manage the method for this injection quality by the amount that is determined at N-acetylneuraminic acid contained in this injection.
(25) management of record contains the method that vaccinia virus is inoculated the injection quality of rabbit inflammation skin extract in (24); wherein; in this injection; the amount of contained N-acetylneuraminic acid is 4800ng when above in the every 1mL of this injection in this injection, and the quality of setting this injection is qualified.
(26) management of record contains the method that vaccinia virus is inoculated the injection quality of rabbit inflammation skin extract in (24); wherein; in this injection; when the amount of contained N-acetylneuraminic acid was 4800～27000ng in the every 1mL of this injection in this injection, the quality of setting this injection was qualified.
(27) management of record contains the method that vaccinia virus is inoculated the injection quality of rabbit inflammation skin extract in (24); wherein; in the injection of the 3mL dress that contains vaccinia virus inoculation rabbit inflammation skin extract; the amount of contained N-acetylneuraminic acid is 14400ng when above in every 1 pipe of this injection in this injection, and the quality of setting this injection is qualified.
(28) management of record contains the method that vaccinia virus is inoculated the injection quality of rabbit inflammation skin extract in (24); wherein; in the injection of the 3mL dress that contains vaccinia virus inoculation rabbit inflammation skin extract; when the amount of contained N-acetylneuraminic acid was 14400～81000ng in every 1 pipe of this injection in this injection, the quality of setting this injection was qualified.
(29) carry out with the method for putting down in writing in any one in (24)～(28) injection that qualitative control obtains.
(30) in the tablet that contains vaccinia virus inoculation rabbit inflammation skin extract, manage the method for this tablet quality by the amount that is determined at N-acetylneuraminic acid contained in this tablet.
(31) management of record contains the method that vaccinia virus is inoculated the tablet quality of rabbit inflammation skin extract in (30); wherein; in this tablet; the amount of contained N-acetylneuraminic acid is 16000ng when above in every 1 of this tablet in this tablet, and the quality of setting this tablet is qualified.
(32) management of record contains the method that vaccinia virus is inoculated the tablet quality of rabbit inflammation skin extract in (30); wherein; in this tablet; when the amount of contained N-acetylneuraminic acid was 16000～90000ng in every 1 of this tablet in this tablet, the quality of setting this tablet was qualified.
(33) carry out with the method for putting down in writing in any one in (30)～(32) tablet that qualitative control obtains.
(34) estimate the method for this extract by the amount that is determined at N-acetylneuraminic acid contained in vaccinia virus inoculation rabbit inflammation skin extract.
(35) estimate the method for said preparation by the amount that is determined at N-acetylneuraminic acid contained in the preparation that contains vaccinia virus inoculation rabbit inflammation skin extract.
(36) estimate the method for this injection by the amount that is determined at N-acetylneuraminic acid contained in the injection that contains vaccinia virus inoculation rabbit inflammation skin extract.
(37) estimate the method for this tablet by the amount that is determined at N-acetylneuraminic acid contained in the tablet that contains vaccinia virus inoculation rabbit inflammation skin extract.
Industrial utilizability
As mentioned above, the invention provides vaccinia virus inoculation rabbit inflammation skin extract or the preparation take this extract as effective ingredient of the N-acetylneuraminic acid that contains ormal weight.In addition, the invention provides by the N-acetylneuraminic acid of guaranteeing to contain ormal weight in such extract or preparation more strictly to this extract and preparation carry out the method for qualitative control or like this operation carry out extract or the preparation that qualitative control obtains.Such extract and preparation can more strictly be guaranteed the constancy of its quality owing to using the bio-tissue manufacturing, so be very useful.
Claims (37)
1. a vaccinia virus is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1 unit contains the above N-acetylneuraminic acid of 4000ng.
2. a vaccinia virus is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1 unit contains the N-acetylneuraminic acid of 4000～22500ng.
3. the vaccinia virus as liquid is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1mL contains the above N-acetylneuraminic acid of 4800ng.
4. the vaccinia virus as liquid is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1mL contains the N-acetylneuraminic acid of 4800～27000ng.
5. one kind contains the preparation that vaccinia virus is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1 unit contains the above N-acetylneuraminic acid of 4000ng.
6. one kind contains the preparation that vaccinia virus is inoculated rabbit inflammation skin extract, it is characterized in that:
Every 1 unit contains the N-acetylneuraminic acid of 4000～22500ng.
7. one kind contains the injection that vaccinia virus is inoculated rabbit inflammation skin extract, it is characterized in that: every 1mL contains the above N-acetylneuraminic acid of 4800ng.
8. one kind contains the injection that vaccinia virus is inoculated rabbit inflammation skin extract, and it is characterized in that: every 1mL contains the N-acetylneuraminic acid of 4800～27000ng.
9. one kind contains the injection that vaccinia virus is inoculated the 3mL dress of rabbit inflammation skin extract, it is characterized in that:
Every 1 pipe contains the above N-acetylneuraminic acid of 14400ng.
10. one kind contains the injection that vaccinia virus is inoculated the 3mL dress of rabbit inflammation skin extract, it is characterized in that:
Every 1 pipe contains the N-acetylneuraminic acid of 14400～81000ng.
11. a tablet that contains vaccinia virus inoculation rabbit inflammation skin extract is characterized in that: every 1 contains the above N-acetylneuraminic acid of 16000ng.
12. a tablet that contains vaccinia virus inoculation rabbit inflammation skin extract is characterized in that: every 1 N-acetylneuraminic acid that contains 16000～90000ng.
13. preparation as described in claim 5 or 6 is characterized in that:
It is analgesics.
14. injection as described in any one in claim 7～10 is characterized in that:
It is analgesics.
15. tablet as described in claim 11 or 12 is characterized in that:
It is analgesics.
16. a method of managing the quality of vaccinia virus inoculation rabbit inflammation skin extract is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described extract.
17. the method for the quality of management vaccinia virus inoculation rabbit inflammation skin extract as claimed in claim 16 is characterized in that:
In every 1 unit of described extract, the amount of the N-acetylneuraminic acid that described extract is contained is 4000ng when above, and the quality of setting described extract is qualified.
18. the method for the quality of management vaccinia virus inoculation rabbit inflammation skin extract as claimed in claim 16 is characterized in that:
In every 1 unit of described extract, when the amount of the N-acetylneuraminic acid that described extract is contained was 4000～22500ng, the quality of setting described extract was qualified.
19. a vaccinia virus inoculation rabbit inflammation skin extract is characterized in that:
It carries out qualitative control with the described method of any one in claim 16～18 and obtains.
20. a management contains the method for quality of the preparation of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described preparation.
21. management as claimed in claim 20 contains the method for quality of the preparation of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In every 1 unit of vaccinia virus in described preparation inoculation rabbit inflammation skin extract, the amount of the N-acetylneuraminic acid that described preparation is contained is 4000ng when above, and the quality of the described preparation of setting is qualified.
22. management as claimed in claim 20 contains the method for quality of the preparation of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In every 1 unit of vaccinia virus in described preparation inoculation rabbit inflammation skin extract, when the amount of the N-acetylneuraminic acid that described preparation is contained was 4000～22500ng, the quality of setting described preparation was qualified.
23. a preparation is characterized in that:
It carries out qualitative control with the described method of any one in claim 20～22 and obtains.
24. a management contains the method for quality of the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is by in described injection, measures the amount of the contained N-acetylneuraminic acid of described injection and carries out.
25. management as claimed in claim 24 contains the method for quality of the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In described injection, the amount of the N-acetylneuraminic acid that described injection is contained is 4800ng when above in the every 1mL of described injection, and the quality of setting described injection is qualified.
26. management as claimed in claim 24 contains the method for quality of the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In described injection, when the amount of the N-acetylneuraminic acid that described injection is contained was 4800～27000ng in the every 1mL of described injection, the quality of setting described injection was qualified.
27. management as claimed in claim 24 contains the method for quality of the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In the injection of the 3mL dress that contains vaccinia virus inoculation rabbit inflammation skin extract, in every 1 pipe of described injection, the amount of the N-acetylneuraminic acid that described injection is contained is 14400ng when above, and the quality of the described injection of setting is qualified.
28. management as claimed in claim 24 contains the method for quality of the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In the injection of the 3mL dress that contains vaccinia virus inoculation rabbit inflammation skin extract, in every 1 pipe of described injection, when the amount of the N-acetylneuraminic acid that described injection is contained was 14400～81000ng, the quality of setting described injection was qualified.
29. an injection is characterized in that:
It carries out qualitative control with the described method of any one in claim 24～28 and obtains.
30. a management contains the method for quality of the tablet of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is by in described tablet, measures the amount of the contained N-acetylneuraminic acid of described tablet and carries out.
31. management as claimed in claim 30 contains the method for quality of the tablet of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
In described tablet, the amount of the N-acetylneuraminic acid that described tablet is contained is 16000ng when above in every 1 of described tablet, and the quality of setting described tablet is qualified.
32. the method for the quality of the tablet that contains vaccinia virus inoculation rabbit inflammation skin extract as claimed in claim 30 is characterized in that:
In described tablet, when the amount of the N-acetylneuraminic acid that described tablet is contained was 16000～90000ng in every 1 of described tablet, the quality of setting described tablet was qualified.
33. a tablet is characterized in that:
It carries out qualitative control with the described method of any one in claim 30～32 and obtains.
34. an evaluation contains the method for vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described extract.
35. an evaluation contains the method for the preparation of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described preparation.
36. an evaluation contains the method for the injection of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described injection.
37. an evaluation contains the method for the tablet of vaccinia virus inoculation rabbit inflammation skin extract, it is characterized in that:
It is to be undertaken by the amount of measuring the contained N-acetylneuraminic acid of described tablet.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2012-225133 | 2012-10-10 | ||
| JP2012225133A JP5275502B1 (en) | 2012-10-10 | 2012-10-10 | Extracts and formulations |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103110663A true CN103110663A (en) | 2013-05-22 |
| CN103110663B CN103110663B (en) | 2016-04-06 |
Family
ID=48409064
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210528626.0A Active CN103110663B (en) | 2012-10-10 | 2012-12-10 | extract and preparation |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP5275502B1 (en) |
| CN (1) | CN103110663B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107635570A (en) * | 2015-05-29 | 2018-01-26 | 日本脏器制药株式会社 | Multipotent stem cells migrate accelerator |
| CN109512838A (en) * | 2017-09-15 | 2019-03-26 | 天津小西生物医药科技有限公司 | A kind of rabbit fur extractive and its preparation method and application |
| CN110179828A (en) * | 2018-02-20 | 2019-08-30 | 学校法人东海大学 | GalNAc transferase expression promotor containing Vaccination with vaccinia virus inflammatory tissue extract |
| CN111511403A (en) * | 2017-12-28 | 2020-08-07 | 学校法人兵库医科大学 | Lipocalin-type prostaglandin D2 synthase production promoter |
| CN113424063A (en) * | 2019-01-30 | 2021-09-21 | 刘军 | Preparation for inhibiting or relieving brain inflammation |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5930419B2 (en) | 2014-03-14 | 2016-06-08 | 株式会社カイジョー | Bonding equipment |
| JP5930423B2 (en) | 2014-05-09 | 2016-06-08 | 株式会社カイジョー | Bonding equipment |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101410124A (en) * | 2006-03-30 | 2009-04-15 | 国立大学法人京都大学 | Thioredoxin production promoting agent |
| CN101528947A (en) * | 2006-09-06 | 2009-09-09 | 日本脏器制药株式会社 | Method for study, determination or evaluation by gene expression analysis |
| CN101790377A (en) * | 2007-08-31 | 2010-07-28 | 国立大学法人九州大学 | Prophylactic or alleviating agent for peripheral nerve disorder induced by anti-cancer agent |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3649989B2 (en) * | 1999-04-16 | 2005-05-18 | 日本臓器製薬株式会社 | Test method |
| JP4005115B1 (en) * | 2007-02-08 | 2007-11-07 | 日本臓器製薬株式会社 | Pain disease treatment |
-
2012
- 2012-10-10 JP JP2012225133A patent/JP5275502B1/en active Active
- 2012-12-10 CN CN201210528626.0A patent/CN103110663B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101410124A (en) * | 2006-03-30 | 2009-04-15 | 国立大学法人京都大学 | Thioredoxin production promoting agent |
| CN101528947A (en) * | 2006-09-06 | 2009-09-09 | 日本脏器制药株式会社 | Method for study, determination or evaluation by gene expression analysis |
| CN101790377A (en) * | 2007-08-31 | 2010-07-28 | 国立大学法人九州大学 | Prophylactic or alleviating agent for peripheral nerve disorder induced by anti-cancer agent |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107635570A (en) * | 2015-05-29 | 2018-01-26 | 日本脏器制药株式会社 | Multipotent stem cells migrate accelerator |
| CN107635570B (en) * | 2015-05-29 | 2019-03-19 | 日本脏器制药株式会社 | Multipotent stem cells migrate promotor |
| CN109512838A (en) * | 2017-09-15 | 2019-03-26 | 天津小西生物医药科技有限公司 | A kind of rabbit fur extractive and its preparation method and application |
| CN109512838B (en) * | 2017-09-15 | 2022-05-10 | 天津小西生物医药科技有限公司 | Rabbit skin extract and preparation method and application thereof |
| CN111511403A (en) * | 2017-12-28 | 2020-08-07 | 学校法人兵库医科大学 | Lipocalin-type prostaglandin D2 synthase production promoter |
| CN110179828A (en) * | 2018-02-20 | 2019-08-30 | 学校法人东海大学 | GalNAc transferase expression promotor containing Vaccination with vaccinia virus inflammatory tissue extract |
| CN110179828B (en) * | 2018-02-20 | 2023-10-31 | 学校法人东海大学 | N-acetylgalactosamine transferase expression promoter containing vaccinia virus inoculated inflammatory tissue extract |
| CN113424063A (en) * | 2019-01-30 | 2021-09-21 | 刘军 | Preparation for inhibiting or relieving brain inflammation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103110663B (en) | 2016-04-06 |
| JP5275502B1 (en) | 2013-08-28 |
| JP2014076961A (en) | 2014-05-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103110663B (en) | extract and preparation | |
| CN103357006B (en) | Method for examining preparation containing extractive | |
| JP6680829B2 (en) | Extract and preparation containing the extract | |
| JP6043922B2 (en) | Extract and formulation containing the extract | |
| JP5490939B2 (en) | Extracts and formulations | |
| JP6412891B2 (en) | Extracts and formulations | |
| JP5844338B2 (en) | Extract and formulation containing the extract | |
| JP5844320B2 (en) | Extract and formulation containing the extract | |
| JP5438234B1 (en) | Extract and formulation containing the extract | |
| JP2018162311A (en) | Extract and formulation | |
| JP2016216518A (en) | Extract and formulation containing the same | |
| JP2014101381A (en) | Extract and preparation |
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 |