CN102430116A - Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer - Google Patents
Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer Download PDFInfo
- Publication number
- CN102430116A CN102430116A CN2011102402782A CN201110240278A CN102430116A CN 102430116 A CN102430116 A CN 102430116A CN 2011102402782 A CN2011102402782 A CN 2011102402782A CN 201110240278 A CN201110240278 A CN 201110240278A CN 102430116 A CN102430116 A CN 102430116A
- Authority
- CN
- China
- Prior art keywords
- coagulation factor
- heat treatment
- blood coagulation
- factor vii
- stabilizing agent
- 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.)
- Pending
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a dry heat treatment method for a human coagulation factor VIII preparation. Before a freeze-drying step, the stabilizer is added into human coagulation factor VIII solution, wherein the stabilizer comprises human serum albumin, Ca2+ soluble salt, amino acid, sodium citrate and sodium chloride; and the concentration percentage of the human serum albumin is 0.5 to 5 percent and the concentration of Ca2+ is 1mmol/L. The invention also discloses the dry heat treatment stabilizer for the human coagulation factor VIII preparation. The dry heat treatment method is performed for 72 hours at 80 DEG C and is effective on both lipid envelop virus and non-lipid envelop virus. Usually, heat treatment is the last step. The method is low in cost and higher in maneuverability and controllability.
Description
Technical field
The present invention relates to medical technical field, more specifically, the present invention relates to a kind of human blood coagulation factor VII I preparation that prevents at 80 ℃, in the xeothermic inactivation process of 72h, a kind of dry heat treatment method and dry heat treatment stabilizing agent that is used for human blood coagulation factor VII I preparation.
Background technology
Human blood coagulation factor VII I (Human Coagulation Factor VIII) claims " antihmemophilic globulin ", is one of important component of intrinsic coagulation system in the body.Factor IX and factors IX a, phospholipid and calcium ion form complex, in the activation process of factor X, play the cofactor effect.Human blood coagulation factor VII I preparation is to prevent and treat the hemorrhage specially good effect Blood Preparations of hemophilia A people.Because the particularity of blood products carries viruses such as AIDS, hepatitis A, hepatitis B easily, if inactivation of virus is not thorough, is easy to cause patient's infection so, so inactivation of virus is a committed step of producing human blood coagulation factor VII I.
Multiple virus inactivating methods such as the present Pasteur's deactivation of report at present, S/D inactivation of virus method, xeothermic deactivation method all have been applied in the production process of the dense preparation of the human blood coagulation factor VII I factor, as: Pasteur's deactivation that the product Humate-P of German Centeon Pharma GmbH company promptly adopts.In above-mentioned virus inactivating method; S/D virus inactivating method reaction condition is relatively gentle; Can not cause the loss of tiring of human blood coagulation factor VII I; Can only be but its action principle has limited it as the deactivation of lipid-coated virus, the thrombin goods of only handling with the S/D method still have the possibility of non-lipid-coated virus such as propagating hepatitis A, B19.Pasteur's deactivation method commonly used at present is comparatively reliable in interior temperature deactivation method, and especially xeothermic deactivation is more stable.
The patent No. is that CN1181974 discloses a kind of low sugar, and is stable, do not contain a kind of recombinant NiaState of white egg.But this recombinant NiaState does not contain the stabilizing agent of human albumin's rFVIII (recombinant human blood coagulation factor VII I), and this recombinant human blood coagulation factor VII I does not adopt xeothermic virus inactivation technology.
But in xeothermic inactivation process, finding is not having under the situation of stabilizing agent, and human blood coagulation factor VII I tires loss very seriously, may cause 50% the loss of tiring at most.This has not only caused a large amount of economic losses, has also increased production cost simultaneously, therefore changes the prescription of human blood coagulation factor VII I, and it is very necessary reducing the loss of tiring.
Summary of the invention
For solving the problems of the technologies described above, the object of the invention is to overcome or improves existing problem, the invention provides a kind of stabilizing agent of dry heat treatment of human blood coagulation factor VII I preparation.
For realizing above-mentioned purpose, technical scheme of the present invention is:
A kind of dry heat treatment method that is used for human blood coagulation factor VII I preparation is characterized in that, before step of freeze drying, in described human blood coagulation factor VII I solution, adds said stabilizing agent, and wherein said stabilizing agent comprises human albumin, Ca
2+Dissolves, aminoacid, trisodium citrate and sodium chloride, wherein said human albumin's percentage is 0.5%-5%, Ca
2+Concentration is 1mmol/L.
Another aspect of the present invention, described human albumin's percentage is 0.5%-2%.
Another aspect of the present invention, described aminoacid are glycine, lysine or its salt.
Another aspect of the present invention before step of freeze drying, in described human blood coagulation factor VII I solution, adds the glycine that described stabilizing agent contains 0.1-10%.
Another aspect of the present invention, before step of freeze drying, in described human blood coagulation factor VII I solution, adding described sodium chloride content is 0.01mmol/L-0.1mmol/L.
Another aspect of the present invention, said method are 80 ℃, xeothermic deactivation under 72 hours conditions.
Another embodiment of the present invention, described stabilizing agent comprises human albumin, Ca
2+Dissolves, aminoacid, trisodium citrate and sodium chloride, wherein said human albumin's percentage is 0.5%-5%, Ca
2+Concentration is 1mmol/L.
Another aspect of the present invention, described human albumin's percentage is 0.5%-2%.
Another aspect of the present invention, described aminoacid are glycine, lysine or its esters, and described stabilizing agent contains the glycine of 0.1-10%.
Another embodiment of the present invention, the application of a kind of dry heat treatment stabilizing agent in human blood coagulation factor VII I preparation.
Human blood coagulation factor VII I is a kind of blood products that uses human normal plasma as raw material, needs the virus that adopts xeothermic virus inactivation technology deactivation to exist.
Beneficial effect of the present invention is:
Stabilizing agent provided by the invention makes preparation at 80 ℃, and in the process of 72 hours dry heat treatment, the loss of human blood coagulation factor VII I reduces, and activity recovery can improve 5%-50%.In human blood coagulation factor VII I preparation, add the above stabilizing agent, can reach the effect of inactivation of virus, can reduce the loss of activity of blood coagulation factor VIII again.
The present invention is chosen in 80 ℃ of 72 hours following dry heat treatment, and is all effective to fat peplos and non-lipid-coated virus, and the last often step of dry heat treatment, and cost is lower, and operability and controllability are stronger.
The specific embodiment
Below in conjunction with instance to stabilizing agent of the present invention to human blood coagulation factor VII I preparation at 80 ℃, detailed explanation is done in the protective effect in 72 hours xeothermic inactivation process.
Embodiment 1: inactivation of virus is verified after adding stabilizing agent
(1) preparation of human blood coagulation factor VII I
Centrifugal blood plasma obtains cryoprecipitate under 0-4 ℃, and the dissolving cryoprecipitate is through Acid precipitation and clarification filtration; Add S/D solution to tween80 final concentration 1%, TNBP 0.3%, and 24-26 ℃ is incubated 6 hours down; Obtain containing the protein solution of human blood coagulation factor VII I then through column chromatography; The gained eluent is with containing 0.01-0.1MNaCl, 0.01M trisodium citrate, 0.1%-0.349% lysine hydrochloride; The dialysis solution of 0.1%-10% glycine carries out ultrafiltration desalination dialysis, obtains the concentrated solution of human blood coagulation factor VII I.
(2) add stabilizing agent, preparation human blood coagulation factor VII I lyophilized preparation
In concentrated solution, adding 20% human albumin concentration to the solution is 2%, adds CaCl
2To Ca
2+Concentration is to 1mmol/L.Degerming, packing is finished product through viral xeothermic deactivation in 80 ℃/72 hours after the lyophilizing.Before the lyophilization.
(3) inactivation of virus
Get three crowdes of human blood coagulation factor VII I respectively and carry out the inactivation of virus experiment, three batches of human blood coagulation factor VII I inactivation of virus experimental results are following:
Table 1 human blood coagulation factor VII I80 ℃ of 72 hours inactivation of virus result
Can find out that by table 1 behind 80 ℃ of 72 hours inactivation of virus, the human blood coagulation factor VII I preparation that adds stabilizing agent meets fully that " requirement of Chinese pharmacopoeia explain that the xeothermic viral inactivation treatment behind the adding stabilizing agent is effective.
Embodiment 2: human blood coagulation factor VII I stabilizing agent is to the influence of preparation in 80 ℃ of xeothermic deactivations in 72 hours
When preparation human blood coagulation factor VII I concentrated solution; Dialysis solution concentrates dialysis with the stabilizing agent of 0.9% glycine and made as dialysis solution respectively; Observe the protective effect of stabilizing agent to the human blood coagulation factor VII I preparation in lyophilizing and the xeothermic deactivation, the response rate of tiring after the lyophilizing and after the xeothermic deactivation is following:
Table 2 human blood coagulation factor VII I stabilizing agent protection experimental result
Can be found out by table 2, not adding stabilizing agent, only contain under the situation of excipient 0.9% glycine that the loss of tiring is up to 40%-50%, after adding stabilizing agent, the loss rate of tiring is higher than 90%.
Embodiment 3: human albumin and Ca in the dialysis solution
2+Protective effect to human blood coagulation factor VII I.
1. prepare human blood coagulation factor VII I by conventional method, remove CaCl
2Outside the human albumin, add stabilizing agent by standard, preparation human blood coagulation factor VII I preparation concentrated solution.
2. observe the activity recovery of redissolution back sample clarity and human blood coagulation factor VII I
With the concentrated solution of method for preparing, the human blood coagulation factor VII I that according to the form below adds stabilizing agent carries out lyophilizing, 80 ℃ of experiments in 72 hours, and the clarity of the solution after the observation, and detect and tire, observe the influence of protective agent to the human blood coagulation factor VII I in the inactivation of virus.The result sees table 3.
Table 3: the result of study of the final lysate of human blood coagulation factor VII I different formulations
Can find out from table 3, redissolve effect better (outward appearance is good, and it is stable to tire) before and after the white lysate secondary inactivation of virus with the calcium chloride composition of people to contain.Wherein with 2% human albumin, the human blood coagulation factor VII I preparation outward appearance that 1mmol/L calcium chloride prescription obtains with tire ideal.
Embodiment 4:NaCl is to the protection experiment of blood coagulation factor VIII preparation
By conventional method, obtain containing the eluent of high concentration NaCl solution.NaCl concentration in the dialysis solution is set at 0 respectively, 25mmol/L, 50mmol/L, 100mmol/L NaCl concentrates respectively and dialyses to dialysis solution NaCl concentration.Degerming, packing, lyophilizing is through 80 ℃ of 72 hours xeothermic inactivation of virus.Redissolve according to the quality inspection rules, mensuration is tired, and observes outward appearance, and the result sees table 4.
Table 4 variable concentrations NaCl is to the influence of the xeothermic deactivation of human blood coagulation factor VII I
Can find out that by last table when not adding sodium chloride, xeothermic loss reaches 25%-30%; Continuous increase along with sodium chloride concentration; Though outward appearance is had certain influence, can't cause product not meet system inspection rules, the loss of tiring simultaneously minimizing progressively.Because system inspection rules limit Na
+Concentration is no more than 160mmol/L, so the final concentration of NaCl is 100mmol/L.
Embodiment 5: glycine concentration is to the protection experiment of blood coagulation factor VIII
By conventional method, obtain containing the eluent of high concentration NaCl solution.Glycine concentration in the dialysis solution is set at 0.3%, 0.6%, 0.9%, 1.2%, 1.5%, 2% respectively.Concentrate respectively and dialyse to dialysate concentration.Degerming, packing, lyophilizing is through 80 ℃ of 72 hours xeothermic inactivation of virus.Redissolve according to the quality inspection rules, mensuration is tired, and observes outward appearance, and the result sees table 5.
Table 5 variable concentrations glycine is to the influence of the xeothermic deactivation of human blood coagulation factor VII I
Glycine is as a kind of excipient, and its concentration has certain influence to the outward appearance of human blood coagulation factor VII I.Carried out the experiment of various concentration respectively.Can find that through table 5 adding of glycine can improve the outward appearance of human blood coagulation factor VII I, but, therefore adopt the concentration of 0.9% glycine in the buffer system as dialysis solution to the not significant protection effect of tiring.
In sum, the behave dry heat treatment of blood coagulation factor VIII of the present invention provides effective stabilizer.
So far invention has been described to have combined embodiment.Be familiar with those skilled in the art and should be appreciated that under situation about not departing from the scope of the present invention with spirit, can easily make various other modifications said embodiment.Therefore, the scope that Rights attached thereto require is not limited to above-mentioned explanation, but broadly construe requirement.
Claims (10)
1. dry heat treatment method that is used for human blood coagulation factor VII I preparation is characterized in that: before step of freeze drying, in described human blood coagulation factor VII I solution, add said stabilizing agent, wherein said stabilizing agent comprises human albumin, Ca
2+Dissolves, aminoacid, trisodium citrate and sodium chloride, wherein said human albumin's percentage is 0.5%-5%, Ca
2+Concentration is 1mmol/L.
2. according to the method described in the claim 1, it is characterized in that: described human albumin's percentage is 0.5%-2%.
3. according to the method described in the claim 1, it is characterized in that: described aminoacid is glycine, lysine or its esters.
4. according to the method described in the claim 3, it is characterized in that: before step of freeze drying, in described human blood coagulation factor VII I solution, add the glycine that described stabilizing agent contains 0.1-10%.
5. according to the method described in the claim 1, it is characterized in that: before step of freeze drying, in described human blood coagulation factor VII I solution, adding described sodium chloride content is 0.01mmol/L-0.1mmol/L.
6. according to the method described in the claim 1, it is characterized in that: said method is at 80 ℃, xeothermic deactivation under 72 hours conditions.
7. dry heat treatment stabilizing agent that is used for human blood coagulation factor VII I preparation, it is characterized in that: described stabilizing agent comprises human albumin, Ca
2+Dissolves, aminoacid, trisodium citrate and sodium chloride, wherein said human albumin's percentage is 0.5%-5%, Ca
2+Concentration is 1mmol/L.
8. according to the stabilizing agent described in the claim 7, it is characterized in that: described human albumin's percentage is 0.5%-2%.
9. according to the stabilizing agent described in the claim 7, it is characterized in that: described aminoacid is glycine, lysine or its esters, and described stabilizing agent contains the glycine of 0.1-10%.
10. according to the application of arbitrary described dry heat treatment stabilizing agent in human blood coagulation factor VII I preparation among the claim 7-9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102402782A CN102430116A (en) | 2011-08-19 | 2011-08-19 | Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102402782A CN102430116A (en) | 2011-08-19 | 2011-08-19 | Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102430116A true CN102430116A (en) | 2012-05-02 |
Family
ID=45978700
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102402782A Pending CN102430116A (en) | 2011-08-19 | 2011-08-19 | Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102430116A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102924562A (en) * | 2012-11-19 | 2013-02-13 | 成都蓉生药业有限责任公司 | Dry heat treatment stabilizer for human blood coagulation factor VIII and application thereof |
| CN103611162A (en) * | 2013-12-11 | 2014-03-05 | 武汉生物制品研究所有限责任公司 | Freeze-drying protective agent for human blood coagulation factor VIII and preparation method of freeze-drying protective agent |
| CN106139127A (en) * | 2016-08-05 | 2016-11-23 | 无锡药明康德生物技术股份有限公司 | Factor Ⅷ,rDNA lyophilized formulations |
| CN107337727A (en) * | 2017-08-03 | 2017-11-10 | 国药集团武汉血液制品有限公司 | A kind of haematogenous human blood coagulation factors VIII preparation method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1181974A (en) * | 1996-07-12 | 1998-05-20 | 美国拜尔公司 | Stabilized albumin-free recombinant factor VIII preparation having low sugar content |
| CN101544683A (en) * | 2008-03-28 | 2009-09-30 | 上海莱士血液制品股份有限公司 | Method and substance for keeping fibrinogen activity in thermal treatment |
| CN101967188A (en) * | 2010-11-08 | 2011-02-09 | 江西博雅生物制药股份有限公司 | Process for preparing human antihemophilic factor VIII |
-
2011
- 2011-08-19 CN CN2011102402782A patent/CN102430116A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1181974A (en) * | 1996-07-12 | 1998-05-20 | 美国拜尔公司 | Stabilized albumin-free recombinant factor VIII preparation having low sugar content |
| CN101544683A (en) * | 2008-03-28 | 2009-09-30 | 上海莱士血液制品股份有限公司 | Method and substance for keeping fibrinogen activity in thermal treatment |
| CN101967188A (en) * | 2010-11-08 | 2011-02-09 | 江西博雅生物制药股份有限公司 | Process for preparing human antihemophilic factor VIII |
Non-Patent Citations (2)
| Title |
|---|
| 曹海军等: "人凝血因子VIII几种冻干保护剂的比较", 《中国输血杂志》, vol. 22, no. 7, 30 July 2009 (2009-07-30) * |
| 沈绮: "第四届血浆蛋白生物技术国际会议简介", 《药物分析杂志》, vol. 25, no. 6, 31 December 2005 (2005-12-31) * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102924562A (en) * | 2012-11-19 | 2013-02-13 | 成都蓉生药业有限责任公司 | Dry heat treatment stabilizer for human blood coagulation factor VIII and application thereof |
| WO2014075435A1 (en) * | 2012-11-19 | 2014-05-22 | 成都蓉生药业有限责任公司 | Dry heat treatment stabilizer for human blood coagulation factor viii and use thereof |
| CN102924562B (en) * | 2012-11-19 | 2014-07-23 | 成都蓉生药业有限责任公司 | Dry heat treatment stabilizer for human blood coagulation factor VIII and application thereof |
| CN103611162A (en) * | 2013-12-11 | 2014-03-05 | 武汉生物制品研究所有限责任公司 | Freeze-drying protective agent for human blood coagulation factor VIII and preparation method of freeze-drying protective agent |
| CN103611162B (en) * | 2013-12-11 | 2015-09-16 | 武汉生物制品研究所有限责任公司 | Human blood coagulation factors VIII freeze drying protectant and preparation method thereof |
| CN106139127A (en) * | 2016-08-05 | 2016-11-23 | 无锡药明康德生物技术股份有限公司 | Factor Ⅷ,rDNA lyophilized formulations |
| CN106139127B (en) * | 2016-08-05 | 2020-04-07 | 无锡药明生物技术股份有限公司 | Recombinant blood coagulation factor VIII freeze-dried preparation |
| CN107337727A (en) * | 2017-08-03 | 2017-11-10 | 国药集团武汉血液制品有限公司 | A kind of haematogenous human blood coagulation factors VIII preparation method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0315968B1 (en) | Plasma and recombinant protein formulations in low ionic strength media | |
| CN102580062B (en) | Human blood coagulation factor VII I and the dry heat treatment stabilizer of vWF complex or human blood coagulation factor VII I preparation | |
| CA1329760C (en) | Plasma and recombinant protein formulations in high ionic strength media | |
| US4876241A (en) | Stabilization of biological and pharmaceutical products during thermal inactivation of viral and bacterial contaminants | |
| JP5719266B2 (en) | Method for stabilizing plasma protein cryoprecipitates for use in virus-inactivated heat treatment | |
| CN104231073B (en) | Preparation method of human coagulation factor VIII | |
| JPH0348170B2 (en) | ||
| HRP931496A2 (en) | Process for the preparation of biological preparations not containing (active) viruses | |
| CN107849086B (en) | Method for producing hepatitis B immunoglobulins derived from plasma | |
| WO2014075435A1 (en) | Dry heat treatment stabilizer for human blood coagulation factor viii and use thereof | |
| CN102430116A (en) | Dry heat treatment method for human coagulation factor VIII preparation and dry heat treatment stabilizer | |
| JPH09503218A (en) | Virus inactivation method in the presence of polyalkylene glycol and pharmaceutical preparation obtained thereby | |
| CN104225601B (en) | Human blood coagulation factor VII I is freezed and dry heat treatment protective agent | |
| JPS6237010B2 (en) | ||
| EP0292003B1 (en) | Stabilzation of biological and pharmaceutical products during inactivation of viral and bacterial contaminants | |
| WO1998030230A1 (en) | Protein-containing compositions and process for producing the same | |
| JP4629831B2 (en) | Virus inactivation method | |
| CN102416171B (en) | Protective agent in process for performing dry heat virus inactivation on high-purity prothrombin complex concentrate products | |
| CN103041380B (en) | Stabilizer in dry heat treatment process of human antithrombase preparation | |
| CN106011116A (en) | Preparation method of human thrombin | |
| CN105879038B (en) | Dry heat treatment stabilizer for preparing human prothrombin complex and application thereof | |
| CN113577295B (en) | Human fibrinogen dry heat treatment stabilizer and application thereof | |
| JPH01305036A (en) | Heat-treatment of plasma protein component and drug preparation containing plasma protein component | |
| EP3624833A1 (en) | C1-esterase inhibitor preparation | |
| RU2785431C2 (en) | C1-esterase inhibitor 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 | ||
| C53 | Correction of patent of invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Yan Chen Inventor after: Xiong Keqiang Inventor before: Yan Chen Inventor before: Xiong Keqiang |
|
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120502 |
|
| RJ01 | Rejection of invention patent application after publication |