CN104940902B - A kind of stablizing solution of polyethylene glycol integrated interferon variant - Google Patents
A kind of stablizing solution of polyethylene glycol integrated interferon variant Download PDFInfo
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- CN104940902B CN104940902B CN201510289225.8A CN201510289225A CN104940902B CN 104940902 B CN104940902 B CN 104940902B CN 201510289225 A CN201510289225 A CN 201510289225A CN 104940902 B CN104940902 B CN 104940902B
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- 108010050904 Interferons Proteins 0.000 title claims abstract description 44
- 102000014150 Interferons Human genes 0.000 title claims abstract description 44
- 229940079322 interferon Drugs 0.000 title claims abstract description 44
- 229920001223 polyethylene glycol Polymers 0.000 title claims abstract description 31
- 239000002202 Polyethylene glycol Substances 0.000 title claims abstract description 25
- 239000000243 solution Substances 0.000 claims abstract description 50
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 19
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 17
- 229930182817 methionine Natural products 0.000 claims description 17
- 229920001214 Polysorbate 60 Polymers 0.000 claims description 16
- 230000006320 pegylation Effects 0.000 claims description 10
- 230000003139 buffering effect Effects 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 4
- 150000002148 esters Chemical class 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 26
- 239000007853 buffer solution Substances 0.000 abstract description 15
- 239000012634 fragment Substances 0.000 abstract description 3
- 125000000487 histidyl group Chemical class [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 abstract 1
- 239000012535 impurity Substances 0.000 description 28
- 230000003647 oxidation Effects 0.000 description 19
- 238000007254 oxidation reaction Methods 0.000 description 19
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 16
- 235000006109 methionine Nutrition 0.000 description 16
- 238000002347 injection Methods 0.000 description 12
- 239000007924 injection Substances 0.000 description 12
- 238000003860 storage Methods 0.000 description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 150000002411 histidines Chemical class 0.000 description 9
- 230000006641 stabilisation Effects 0.000 description 9
- 238000011105 stabilization Methods 0.000 description 9
- 230000015556 catabolic process Effects 0.000 description 8
- 238000006731 degradation reaction Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 7
- 239000003223 protective agent Substances 0.000 description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000000872 buffer Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 108010047761 Interferon-alpha Proteins 0.000 description 5
- 102000006992 Interferon-alpha Human genes 0.000 description 5
- 229910019142 PO4 Inorganic materials 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 5
- 239000010452 phosphate Substances 0.000 description 5
- 239000003381 stabilizer Substances 0.000 description 5
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 239000003963 antioxidant agent Substances 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000010525 oxidative degradation reaction Methods 0.000 description 4
- 230000002035 prolonged effect Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- -1 sorbierite Substances 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 230000001133 acceleration Effects 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 2
- 229940090438 infergen Drugs 0.000 description 2
- 108010010648 interferon alfacon-1 Proteins 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical class CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- NSENZNPLAVRFMJ-UHFFFAOYSA-N 2,3-dibutylphenol Chemical compound CCCCC1=CC=CC(O)=C1CCCC NSENZNPLAVRFMJ-UHFFFAOYSA-N 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102100026720 Interferon beta Human genes 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 235000001630 Pyrus pyrifolia var culta Nutrition 0.000 description 1
- 240000002609 Pyrus pyrifolia var. culta Species 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 159000000021 acetate salts Chemical class 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 229940102223 injectable solution Drugs 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 150000002742 methionines Chemical class 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229940068977 polysorbate 20 Drugs 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229940068968 polysorbate 80 Drugs 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
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- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
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- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to a kind of polyethylene glycol integrated interferon variant stablizing solutions, using histidine salt as buffer system, by controlling pH 5.0 5.5, can effectively control the problem of segment is broken in preparation, 18 months content of fragment are no more than 0.5% at 4 DEG C.
Description
Technical field
The present invention relates to a kind of stablizing solution of polyethylene glycol integrated interferon variant, i.e., a kind of injection system of stabilization
Agent belongs to technical field of medicine.
Background technology
Interferon be by virus function inactivating or living after permissive cell, generated by permissive cell genome encoding
One group of glycoprotein.Its activity and antigenicity all depend on the protein in molecule, and unrelated with its glycosyl.According to its source and
IEN points can be IFN-α, IFN-β, IFN-γ, IFN- λ s by structure, they are respectively by leucocyte, fibroblast and activation T
Cell generates.IFN-α is polygenes product, has more than ten to plant different subtype, but their bioactivity is essentially identical.IFN, which is removed, to be had
Outside antivirus action, also antitumor, immunological regulation, control cell Proliferation and cause fever the effects that.It clinically uses at present
Interferon obtained mostly by gene recombination technology.
To improve the activity of natural interferon, Amgen companies of the U.S. have synthesized a kind of non-natural interferon for the first time
(Consensus interferon, integrated interferon, trade name Infergen).Infergen is a kind of through computer optimum organization
Non-natural recombinantα-interferon, sequence is formed by 20 kinds of natural alpha-interferon subtype sequences optimum organizations, antiviral
Activity improves 5~10 times than natural interferon.
As other albumen, usually limited by some disadvantages using alpha interferon as drug, including antigenicity and
Half-life period is shorter, and antigenicity leads to the formation of neutralizing antibody and the loss of clinical response, and half-life period is shorter to be intended to
Frequent drug administration is with the treatment effective concentration of Protein requirement.These problems can be by by interferon and polymer such as polyethylene glycol
It is conjugated to overcome.However, although interferon-polyethylene glycol conjugate is clinically effective, such conjugate is clinical real
Being widely used in trampling, it is desirable to be able to it is stabilized within one storage and transportation time, with convenient when in use safe and effective, because
The stabilization formulations conducive to storage are made in this needs.
Composition injection, which is conjugated, in the open glycol interferon alpha of United States Patent (USP) 6180096 can lead to PEG and be done in storage
Disturb the change of plain α properties and conjugated degree.These changes include conjugate degradation (or " structural break ") into Free PEG and interference
Plain α, then free PEG is connected on another glycol interferon molecule or PEG molecules are from a position of conjugate
Another site of point through intramolecular Displacement transfer to identical intramolecular, it is that PEG interferon-' alpha 's are conjugated to solve this degradation problem
Lyophilized preparation is made in object.WO1999/048535, WO2006/020720 also disclose that a kind of similar with lyophilized form solution drop
The scheme of solution problem.
The amino acid sequence of Pegylation recombined human integrated interferon variant has particularity (see patent
ZL01102915.3), several amino acid (GSGGG) are added in conventional integrated interferon N-terminal, and by terminal amino group
The amino of sour (glycine, G) carries out the pegylation of single site, finds that structural break is usual to the research of its solution
It is happened in the GSGGG sequences of the N-terminal of above-mentioned addition (including at PEG connection sites).In patent CN201110189858.3
A kind of Pegylation integrated interferon variant injection is disclosed, prescription includes buffer solution, sorbierite, polysorbate
80, pH 4.5-7.5, wherein in being broken segment research, what storage process was stablized the most is with phosphate buffer (buffering
PH is 6.5-7.5) the stable injectable solution that configures, the formula free IFN-SA segments after structural break during storage
Also 5.4% or more.
However, influencing the factor of the special injection stability of Pegylation recombination integrated interferon variation, not only only
Having structural break, this is a kind of, and interferons product occurs oxidative degradation and has many reports, and Pegylation recombined human is integrated dry
It disturbs and contains several methionines (M) in the amino acid sequence of plain variant simultaneously, therefore oxidative degradation may theoretically occur.
And it is confirmed in long-term or acceleration for stabilization Journal of Sex Research, the increase of oxidation impurities is also the main reason for injection purity declines.
Obviously, in the factor for having numerous influence injections to stablize, the free segment only generated after structural break there is
So many degradation fragment is clearly very unfavorable to the quality of product, it is therefore desirable to provide one kind in prolonged storage more
Integrated interferon variant injection is recombinated for stabilization, the less Pegylation of degradation impurity content.
Goal of the invention
The object of the present invention is to provide a kind of polyethylene glycol weights of the more few stabilization of degradation impurity in prolonged storage
Group integrated interferon variant injection.
Researcher it has surprisingly been found that be buffer with histidine salt, and the pH of solution is controlled during preparation research
System is below 6.0 (such as 4.5-6.0), more preferably by pH controls below 5.5 (such as 5.0-5.5), in formulation storage process
In can more effectively control the formation of fracture segment and oxidative degradation impurity in preparation.
The preparation prescription for the stabilization that the present invention describes, can be by split pieces in prolonged storage (at 4 DEG C, 18 months)
Section control is 5% hereinafter, preferred be controlled at 3% hereinafter, 1% hereinafter, 0.5% hereinafter, more preferably controlling
0.1% hereinafter, being significantly reduced relative to the ejection preparation content of fragment in 201110189858.3 patents, to the quality of preparation
It can significantly improve.In addition, oxidation impurities can also be controlled 3% hereinafter, more preferably control 2% once, most preferably
Control is below 1.5%.
In particular it relates to a kind of stablizing solution formula of polyethylene glycol integrated interferon variant, the stabilization
Solution includes：Polyethylene glycol integrated interferon variant, histidine, protective agent and the stabilizer of therapeutically effective amount, the solution
PH be 4.5-6.0, preferably 5.0-6.0.
In stablizing solution of the present invention, the concentration of the histidine can be any range of 5-50mmol/L, it is preferable that 5-
The purpose of the present invention can be realized in the histidine of 10mmol/L, although greater concentrations of histidine may be more advantageous to polyethylene glycol
The stabilization of integrated interferon injection, but the concentration of bigger realizes that more stable effect less significantly, can but increase system
Agent cost.
In the present invention, protective agent can be selected from carbohydrate, such as sorbierite, sucrose, glucose, mannose, glucan, seaweed
Sugar or sodium chloride, preferably sorbierite, sucrose or trehalose, more preferable sorbierite, protectant concentration can be 1%-
15% (weight/volume, g/100ml solution).
In the present invention, the stabilizer is selected from poly yamanashi esters, preferably poly- such as polysorbate 20 or polyoxyethylene sorbitan monoleate
Sorb ester 80, a concentration of 0.001%-0.01% (weight/volume, g/100ml solution) of the stabilizer.
In the present invention, the pH of the stablizing solution is 4.5-6.0, preferably 5.0-6.0, more preferably 5.0-5.5.
Further, it is preferably to control polyethylene glycol integrated interferon variant solution in prolonged storage because of oxygen
Change and quality caused to decline, stablizing solution of the present invention can also include it is at least one selected from methionine, sodium thiosulfate,
The antioxidant of sodium sulfite, sodium hydrogensulfite, EDTA, vitamin C or dibutylphenol, preferably methionine, the antioxygen
A concentration of 0.01%-0.5% (weight/volume, g/100ml solution) of agent.
In one embodiment of the present invention, it is molten to be related to a kind of polyethylene glycol integrated interferon variant formed as follows
Liquid, including：The Pegylation integrated interferon variant of 50-500 μ g/ml, histidine, the w/v of 5-50mmol/L
Polyoxyethylene sorbitan monoleate that sorbierite, w/v for 1%-10% are 0.001%-0.005%, w/v are
The pH of the methionine of 0.05%-0.15%, the stablizing solution is 5.0-6.0.
In one embodiment of this invention, it by being investigated to different protective agents, finds when selecting sodium chloride, is detrimental to
Preparation stabilization, there are more fracture segments during storage, sorbierite and sucrose are relatively less.
In another embodiment of the present invention, the influence for selecting different buffer systems to evaluate it to preparation stability,
Under the conditions of same pH, it is found that acetate salt buffer system is unfavorable for controlling the crack conditions of preparation, and phosphatebuffer buffer system
Lacking has more oxidation impurities to be formed, (the alternatively referred to as histidine buffer system, by with salt however, histidine buffer system
Acid or sodium hydroxide regulation and control final preparation pH value) in preparation segment fracture and oxidation impurities control advantageously.
In another embodiment of the present invention, choose histidine salt as the buffer system of preparation after, by investigate not
Preparation stability is influenced with pH value, finds high pH (such as 6.7 or 7.2 or more), to the segment fracture of preparation and oxidation impurities
Control it is all unfavorable, and pH controls 6.0 or hereinafter, segment in preparation can be controlled more effectively are being broken and are aoxidizing miscellaneous
The formation of matter.Preferably, when controlling pH 5.5, again advantageously to the stability of preparation.
In a more specific embodiment, it is related to a kind of polyethylene glycol integrated interferon variant solution formed as follows,
Including：Stablizing solution according to any one of claims 8, the solution include the Pegylation integrated interferon variation of 150 μ g/ml
Polyoxyethylene sorbitan monoleate that sorbierite that body, the histidine of 10mmol/L, w/v are 4%, w/v are 0.004%,
The pH of the methionine that w/v is 0.1%, the stablizing solution is 5.0-5.5.
In the present invention, the preparation method of the stablizing solution is also related to, including：
(a) histidine, sorbierite, methionine, polyoxyethylene sorbitan monoleate are dissolved in the water obtained first solution；
(b) polyethylene glycol integrated interferon variant stoste is dissolved in histidine buffering liquid and the second solution is made；
(c) the first solution and the second solution are uniformly mixed；And
(d) pH of mixed solution in (c) step is adjusted to 5.0-5.5.
In the present invention, histidine salt buffer or histidine buffering liquid are used interchangeably, and use histidine as buffer solution,
In configuration process, finally need to adjust the pH value needed for buffering using sodium hydroxide and/or hydrochloric acid.
Description of the drawings
Fig. 1,2 respectively different stabilizers are to split pieces section and oxidation in polyethylene glycol integrated interferon variant injection
The influence of impurity.
Fig. 3,4 to split pieces section in polyethylene glycol integrated interferon variant injection and aoxidize miscellaneous for different buffer systems
The influence of matter.
Fig. 5,6 is, using histidine as buffer system, condition of different pH are to polyethylene glycol integrated interferon variant injections
The influence of middle split pieces section and oxidation impurities.
Specific implementation mode
Illustrate beneficial effects of the present invention from specific embodiment below, it should be pointed out that cited implementation
Example is not meant to limit the present invention, and those skilled in the art combine the prior art under the introduction of technical solution of the present invention
In Conventional wisdom, be easy to make different transformation.
The present invention is by designing different prescriptions, including different buffer systems, different protein stabilisers, the conditions such as different pH,
The stability of (40C) under acceleration environment is investigated, the situation of change of impurity and oxidation impurities is broken in more different prescriptions, to sieve
The best prescription of both degradations can be controlled by choosing.Due to macromolecular drug mechanism of degradation at different temperatures and dynamics
It often differs, therefore after filtering out candidate prescription under acceleration conditions, needs to carry out long-time stability to investigate to confirm control
Effect.
The different protective agents of embodiment 1 influence polyethylene glycol integrated interferon variant stability of solution
Formulation (table 1)
| Buffer system | Protective agent | Antioxidant | Surfactant | pH | |
| Prescription 3 | Phosphate | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
| Prescription 4 | Phosphate | Sucrose | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
| Prescription 5 | Phosphate | Sodium chloride | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
Oxidation impurities and fracture segment the result is shown in Figure 1 and Fig. 2.From Fig. 1,2 as can be seen that oxidation impurities are protected in by prescription
The influence of agent is smaller, and be broken containing the increase in sodium chloride prescription faster than other prescriptions.
The different buffer systems of embodiment 2 influence polyethylene glycol integrated interferon variant stability of solution
Formulation (table 2)
| Buffer system | Protective agent | Antioxidant | Surfactant | pH | |
| Prescription 3 | Phosphate | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
| Prescription 6 | Acetate | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
| Prescription 7 | Histidine salt | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
The buffer system of prescription is different to oxidation impurities and fracture impurity effect, and oxidation impurities increase in phosphate prescription
Add most soon, oxidation impurities increase is most slow (Fig. 3) in histidine salt prescription；And the fracture impurity increase most block in acetate prescription,
It is relatively slow (Fig. 4) that impurity is broken in phosphate and histidine prescription, comprehensive oxidation impurities and fracture segment consider, selection group
Propylhomoserin salt is as buffer system most beneficial for the stability of polyethylene glycol integrated interferon variant solution.
The different pH of 3 same buffer system of embodiment influence polyethylene glycol integrated interferon variant stability of solution
Formulation (table 3)
| Buffer system | Protective agent | Antioxidant | Surfactant | pH | |
| Prescription 2 | Histidine salt | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 7.2 |
| Prescription 8 | Histidine salt | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.7 |
| Prescription 3 | Histidine salt | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 6.0 |
| Prescription 9 | Histidine salt | Sorbierite | Methionine | Polyoxyethylene sorbitan monoleate | 5.5 |
The pH of prescription has apparent influence to oxidation impurities and fracture impurity, and with the reduction of pH, oxidation and fracture are miscellaneous
The increase of matter all obviously slows down, but to the no significant difference of influence of fracture impurity both when pH is reduced to 6.0 or 5.5
(Fig. 5,6) show that pH plays a crucial role the degradation of Pegylation integrated interferon in control prescription.
Embodiment 4 prepares stable polyethylene glycol integrated interferon variant stablizing solution
The preparation method of preparation
Histidine is dissolved in the water, and sorbierite, methionine, polyoxyethylene sorbitan monoleate are added while stirring, uses hydrogen-oxygen
Change sodium and/or hydrochloric acid and pH is adjusted to desired value, then the prepared solution that obtains is filtered by 0.22 μm of filter, is prepared
First solution；
The polyethylene glycol integrated interferon variant stoste for taking assay approval is prepared with appropriate histidine buffering liquid mixing
Obtain the second solution；
By the first solution and the second solution mixing, it is used in combination sodium hydroxide and/or hydrochloric acid tune pH to desired value, be sterile filtered,
It dispenses to obtain the final product.
Upper all operations carry out in gnotobasis.
By the stablizing solution being prepared in 2-8 DEG C of preservation.
Preparation composition is following (table 4)：
5 preparation stability of embodiment is investigated
It analyzes the content of preparation break sliver section and the content of oxidation impurities, preparation is placed 18 months under the conditions of 4 DEG C, made
Oxidation impurities are detected with Reversed phase high performance liquid chromatography, use the segment of gel chromatography efficient liquid phase detection of broken.And it is surveyed under equal conditions
The case where determining fracture and the oxidation impurities of the preparation prescription A in 201110189858.3 embodiment 4 of patent.
18 months fracture segments are placed under the conditions of 4 DEG C of 5 room temperature of table to investigate
18 months oxidative degradation impurity is placed under the conditions of 4 DEG C of 6 room temperature of table to investigate
It can be seen that after 4 DEG C of long term storages 18 months from upper table (table 5, table 6) result, every stability indicator is apparent
The optimal prescription in patent 201110189858.3 (the preparation A i.e. in patent Example 4) will be better than.
Claims (4)
1. a kind of polyethylene glycol integrated interferon variant stablizing solution comprising the integrated interference of the Pegylation of 150 μ g/ml
The poly- sorb that sorbierite that plain variant, the histidine of 10mmol/L, w/v are 4%, w/v are 0.004%
Ester 80, w/v be 0.1% methionine, and the pH of the stablizing solution be 5.0-5.5, wherein it is described stablize it is molten
After 4 DEG C of liquid is placed 18 months, the integrated interferon variant segment broken to form is no more than 1%.
2. stablizing solution according to claim 1 is broken to form wherein after 4 DEG C of the stablizing solution is placed 18 months
Integrated interferon variant segment is no more than 0.5%.
3. stablizing solution according to claim 2 is broken to form wherein after 4 DEG C of the stablizing solution is placed 18 months
Integrated interferon variant segment is no more than 0.1%.
4. a kind of method preparing stablizing solution described in any one claim in claim 1-3, including：
(a) histidine, sorbierite, methionine, polyoxyethylene sorbitan monoleate are dissolved in the water and the first solution is made；
(b) polyethylene glycol integrated interferon variant stoste is dissolved in histidine buffering liquid and the second solution is made；
(c) the first solution and second of solution are mixed；And
(d) pH of mixed solution in (c) step is adjusted to 5.0-5.5.
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| WO2008062481A2 (en) * | 2006-11-24 | 2008-05-29 | Cadila Healthcare Limited | Formulations of peg-interferon alpha conjugates |
| CN102266550A (en) * | 2011-07-08 | 2011-12-07 | 北京凯因科技股份有限公司 | Polyethylene glycol-consensus interferon mutant injection |
| CN103619324A (en) * | 2011-06-03 | 2014-03-05 | 株式会社Lg生命科学 | Stable liquid formulation of etanercept |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2008062481A2 (en) * | 2006-11-24 | 2008-05-29 | Cadila Healthcare Limited | Formulations of peg-interferon alpha conjugates |
| CN103619324A (en) * | 2011-06-03 | 2014-03-05 | 株式会社Lg生命科学 | Stable liquid formulation of etanercept |
| CN102266550A (en) * | 2011-07-08 | 2011-12-07 | 北京凯因科技股份有限公司 | Polyethylene glycol-consensus interferon mutant injection |
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| 不同添加剂对重组α干扰素溶液稳定性的影响;陈文琼 等;《中国新药杂志》;20081231;第17卷(第16期);1425-1428 * |
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