CN101491682A - PEG-IFN omega conjugate and preparation technique thereof - Google Patents
PEG-IFN omega conjugate and preparation technique thereof Download PDFInfo
- Publication number
- CN101491682A CN101491682A CNA2008103014034A CN200810301403A CN101491682A CN 101491682 A CN101491682 A CN 101491682A CN A2008103014034 A CNA2008103014034 A CN A2008103014034A CN 200810301403 A CN200810301403 A CN 200810301403A CN 101491682 A CN101491682 A CN 101491682A
- Authority
- CN
- China
- Prior art keywords
- conjugate
- recombinant human
- polyethylene glycol
- rhifn
- human interferon
- 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
Images
Abstract
The invention discloses a pegylation recombinant interferon omega conjugate and a preparation process thereof. Compared with an unmodified interferon omega, the pegylation recombinant interferon omega conjugate has longer half-life in a human body, can obviously reduce the number of medication, reduce toxic and side effect, is particularly suitable for treating chronic viral infection sensitive to the interferon omega, and has wide application prospect clinically.
Description
Technical field
Relate to the medicine for treating viral infections preparation of a kind of treatment, especially a kind ofly can obviously reduce the medication number of times interferon ω sensitivity, the Pegylation recombinant human interferon alpha 2 conjugates with long-acting with and preparation technology.
Background technology
Interferon is one group of reactive protein (mainly being glycoprotein) with multiple function, is a kind of cytokine that is produced by mononuclear cell and lymphocyte.They have the broad-spectrum antiviral, influence cell growth and multiple biological activitys such as differentiation, adjusting immunologic function on allogenic cell.Amino acid structure, antigenicity and cell source according to ifn protein can be divided into it: IFN-α, IFN-β, IFN-γ.IFN-ω belongs to IFN-α family, and its structure and size are variant slightly with other IFN-α, but antigenicity has bigger difference.Since the eighties, many studies show that, interferon also has tangible cell proliferation effect except that having antiviral, immunoregulatory effect.
ω-interferon (interferon omega, IFN-ω) be one roughly the same interferon-ALPHA the new interferon molecule of 60% homology is arranged on gene order, have broad-spectrum antiviral, cell proliferation and immunoregulatory function, omega interferon is the I type interferon that Austrian scientist Rudelf Hamptman equals discovery in 1985, it is close with the interferon-alpha function, but the antigenicity difference.In recent years, it is similar that research worker finder's omega interferon has same interferon-alpha, IFN-and tumor necrosis factor, effects such as stronger antiviral duplicates, cell proliferation, and its clinical practice launches gradually.It is clinical that the clinical trial that is used for the treatment of third hepatitis as U.S. Biomedicines company recombinant human omega interferon has entered the II phase.
Pharmaceutical grade protein has become the focus of current drug development research, and the status in clinical treatment becomes more and more important.But the problem of pharmaceutical grade protein maximum is to have the restrictions such as immunogenicity that self stability is poor, the half-life short and may exist.And adopt polyethyleneglycol modified protein technology, can effectively solve this type of defective of pharmaceutical grade protein.
(polyethleneglycol is the polymer of a kind of safety non-toxic, non-activity PEG) to Polyethylene Glycol, is usually used in protein drug and modifies.Davis, Abuchowski and colleague thereof research and develop out the Pegylation technology at first in the seventies in 20th century.Studies show that Pegylation can change pharmacokinetics, and then can change the pharmacodynamics of treatment molecule.U.S. FDA has been ratified several pegylated protein medicines at present, comprising glycol interferon alpha.A large amount of clinical researches show it no matter be single with or share the curative effect of treatment chronic hepatitis C apparently higher than IFN-α with virazole without structural modification.
Clinically, the glycol interferon of extensive use comprise the exploitation of Schering Plough company glycol interferon alpha-2b (trade name, wear happy can, PegIntron); (trade name, Pai Luoxin Pegasys), are widely used in chronic type b and therapy for hepatitis C by uniting with ribavirin to the glycol interferon alpha-2a of Roche Holding Ag's exploitation.
The present invention is directed to the defective that adopts pharmaceutical grade protein self to exist, adopt the Pegylation technology to modify omega interferon, provide a kind of peg molecule to combine, and recombinant human interferon alpha 2 molecule is in conjunction with the preparation method of the Pegylation recombinant human interferon omega medicine of a peg molecule with the N-terminal of recombinant human interferon omega.Effectively improve pharmaceutical grade protein self stability, prolong half-life weak point and reduce immunogenicity, change pharmacokinetics, thereby changed the pharmacodynamics of medicine, aspect clinical effect, have good application prospects.
Summary of the invention
Primary technical task to be solved by this invention provides and a kind ofly helps suitability for industrialized production and can obviously reduce the medication number of times, be used for the Pegylation recombinant human interferon alpha 2 medicine of chronic hepatitis clinical treatment.
The present invention relates to a kind of interferon conjugate of chemical modification, its structure is shown in the I formula:
R-L-M-NH-(rhIFN-ω) I
Wherein, R represents to have C1 ~ water-soluble polymer of 4 alkane end groups,
L represents O, N, NH atom,
M represents C1 ~ 6 aliphatic saturated hydrocarbon subunits,
NH represents the N-terminal amino of rhIFN-ω,
RhIFN-ω represents human recombinant interferon ω.
Wherein water-soluble polymer comprises Polyethylene Glycol, polypropylene glycol, polylactic acid, polymeric amino acid, branch's Polyethylene Glycol, branch's polypropylene glycol, branching polymerization aminoacid etc., has the molecular weight between 5kDa ~ 100kDa, preferred water-soluble polymer is a Polyethylene Glycol, has the molecular weight between 5kDa ~ 40kDa; Preferred Polyethylene Glycol is that an end is the Polyethylene Glycol of methyl protection; L preferably is selected as O atom or NH atom described in the structural formula I formula; The preferred C3 of wherein said M ~ 4 aliphatic saturated hydrocarbon subunits, optimum selection is the straight chain propylidene.
The preparation method of the conjugate that patent of the present invention is mentioned is that the salinity scope of control buffer system is between 10 ~ 100mmol/L, between preferred 50 ~ 80mmol/L; The pH value scope is between 4.0 ~ 9.0, between preferred 4.5 ~ 6.5; Add recombinant human interferon omega and react with the polyethylene glycol aldehyde radical derivative, the molal weight of controlling two reactants than scope between 0.25 ~ 20, between preferred 1.0 ~ 10; Reaction time range is between 1～48 hour, between preferred 8～24 hours; Range of reaction temperature is between 4 ℃～37 ℃, between preferred 4 ℃ ~ 25 ℃; And the NaCNBH3 of adding catalytic amount is as catalyst, product is through too in sieve chromatograph, ion exchange chromatography, hydrophobic chromatography separation and purification, collect Pegylation recombinant human interferon omega that peg molecule connects a recombinant human interferon omega molecule through filtration sterilization, after the lyophilization promptly.Require a kind of pharmaceutical composition that can be used for clinical treatment simultaneously, wherein contain I formula conjugate and medicinal diluent, adjuvant or the carrier of effective dose.
Another technical task to be solved by this invention provides a kind of above-mentioned Pegylation recombinant human interferon alpha 2 medicine production technology.
The synthetic method of conjugate is known in the art shown in the structural formula I, in the presence of sodium cyanoborohydride, and the PEG meeting and the primary amine generation reduction amination of band aldehyde radical.Aldehyde radical is different with other close electroactive group, it only and amido react.Though the reactivity ratio NHS active ester of aldehyde radical is low, it has the reaction condition gentleness, is easy to make PEG to be connected with the surface of protein or other material.Therefore, under lower pH, mPEG-aldehyde can carry out selective reaction to proteinic N end.This reaction condition is between pH value 4.0-6.5, under the buffer system of 50-100mmol/L salinity; Temperature is between 4-25 ℃, and the response time is between 8-24 hour; Reaction mol ratio PEG: rhIFN-ω is 1: 1-10: between 1, generally speaking, select can obtain the end modified product of N-under low temperature, low pH value, the response time suitable condition, and reaction conversion ratio is higher.
Conjugate after the modification can adopt the cation-exchange chromatography purification.For example, behind the mixture upper prop, with the sodium acetate drip washing of pH=4.0 20mM; Reuse 0-1M sodium chloride gradient elution, pH value can be controlled between the 3-5.5, and the best is pH4.0.The component of collecting can use methods such as mass spectrum, SDS-PAGE to confirm by the detection molecules amount.
The factor that influences the PEGization reaction mainly contains 1) pH value; 2) temperature; 3) response time; 4) albumen and PEG reagent mol ratio; 5) protein concentration.By controlling one or more reaction factors, we can obtain desired modification sample
Reaction condition
Polymer molecular weight can be any size, and the peg molecule of structure, selected molecular weight can be that 1kDa is to about 100kDa (term approximately means in the Polyethylene Glycol goods, and some molecules are greater than described molecular weight, and other are less than described molecular weight).Choosing of PEG molecular weight depended on required therapeutic scheme (convenience of for example required slow-release time, activity, processing, antigenicity size or shortage and some other PEG are to the known effect of therapeutic protein and analog thereof).In general, the molecular weight of polymer is big more, and the polymer molecule number that is attached on the protein is few more.Polymer molecular weight high more (branch is many more), polymer should be high more to proteinic ratio.
Reaction rate of charge peg molecule is the same with their concentration in reactant mixture to the ratio of protein molecule, can change, in general, Polyethylene Glycol will be measured according to selected molecular weight polyethylene glycol the optimal proportion (refer to reaction efficiency, promptly do not have unnecessary unreacted protein and polymer molecule in the reactant mixture) of protein molecule.
PH can have influence on the ratio of all polymer molecule and protein molecule, and in general, if reactant liquor pH is lower than pK, the polymer molecule ratio should substantially exceed protein molecule for well; If pH is higher than pK, then polymer is unnecessary too big to the ratio of protein molecule.The difference of pH value of reaction system will cause polymer and combination of proteins site that significant difference is arranged.For PEG-aldehyde modifying protein-terminal amino acid optionally, will make pH value in the reaction system be higher than the pK value of α amino and be lower than the pK value of ε amino, just can reach ideal modification effect.
The response time extent of reaction in time prolongation and increase, relevant with reaction temperature.Typical temperature is high more, and the response time is short more, generally selects hour do not wait half an hour to 24.
Common polymer of reaction temperature and proteins react temperature are 4 degree or 25 degree room temperatures, be reflected at and carry out very slowly under the low temperature state, but help the stable of protein structure, obtaining identical modified outcome needs the time longer, high temperature then helps the carrying out that react, and correspondingly shortens the response time.Also can select a certain temperature conditions between 4 degree-25 degree room temperatures for use.
The separation and purification gel filtration chromatography is come separation component according to the molecular weight size, and therefore the protein after the modification can will be modified with the albumen of unmodified with gel filtration chromatography and effectively separate because molecular weight increases.Cation chromatography technology can be used as effective separation method according to the electric charge difference and separates modified mixture.For example, after cation dress post is good, with 20mM sodium acetate (pH4.0) flushing, carry out linear eluting with the buffer that contains 0-1M sodium chloride then, the pH of buffer can be between 3-3.5, preferred pH=4.0.Can differentiate its molecular weight with traditional method by the chromatography component that the cation chromatography obtains, as: mass spectrum, SDS-PAGE electrophoresis, or the method for other known authenticating compound molecular weight.
The coupling of rhIFN-ω and PEG aldehyde can adopt following method to carry out:
1) provides the buffer system of the salinity of pH 4.0～9.0,10～100mmol/L;
2) recombinant human interferon alpha 2 and polyethylene active ester reacted in the system that step 1) provides 1～48 hour, and temperature is controlled at 4～37 ℃, and proportion control is between 1～10;
3) through step 2) product that obtains of reaction is through behind the ion-exchange chromatography or behind the molecular sieve chromatography chromatography, collects the Pegylation recombinant human interferon alpha 2 eluting peak that mainly contains a peg molecule, is used for purity and active the detection.
The present invention relates to a kind of rhIFN-ω with the PEG pointed decoration, its conjugate structure does not have in prior art.Conjugate of the present invention can significantly increase the half-life of rhIFN-ω, increases the therapeutic effect of rhIFN-ω.
Compared with prior art, outstanding substantive distinguishing features that the present invention possessed and significant technological progress.
Mainly show following 3 aspects:
The advantage of Pegylation recombinant human interferon alpha 2 provided by the present invention-ω medicine provides in vivo the blood levels of longer time, has significantly long-lastingly, reduces frequency injection and makes the patient acceptant;
Recombinant human interferon alpha 2-ω of the present invention adopts engineered method, uses the patent carrier of this research department, use Pichia anomala expression, and through fermenting, obtain behind the purification, purity is greater than 98%, and keeps higher biological activity;
The present invention adopts the method for modifying high to the N-terminal selectivity of recombinant human interferon alpha 2-ω, the N-terminal that is polyethylene active aldehyde and recombinant human interferon omega is connected by covalent, the homogeneity of product is greatly enhanced, easy purification, the activity of purified product can keep preferably.
Description of drawings
Fig. 1 rhIFN-ω and 20K mPEG propionic aldehyde different mol ratio example modification reaction result's SDS-PAGE figure
Wherein swimming lane M, I, A, B, C are respectively:
M.Marker
I.rhIFN-ω
A.rhIFN-ω: mPEG propionic aldehyde=1: 1
B.rhIFN-ω: mPEG propionic aldehyde=1: 3
C.rhIFN-ω: mPEG propionic aldehyde=1: 5
Fig. 2 rhIFN-ω and 20K mPEG butyraldehyde different mol ratio example modification reaction result's SDS-PAGE figure
Wherein swimming lane M, I, A, B, C are respectively:
M.Marker
I.rhIFN-ω
A.rhIFN-ω: mPEG butyraldehyde=1: 1
B.rhIFN-ω: mPEG butyraldehyde=1: 3
C.rhIFN-ω: mPEG butyraldehyde=1: 5
Figure 32 0K mPEG propionic aldehyde and rhIFN-ω reactant gel chromatogram
Figure 42 0K mPEG propionic aldehyde and rhIFN-ω reactant ion exchange chromatography figure
Each component peaks SDS-PAGE figure of Fig. 5 ion exchange chromatography
Wherein P1 is that two PEG modify rhIFN-ω; P2 is that single PEG modifies rhIFN-ω; P3 is rhIFN-ω; I is rhIFN-ω; P modifies rhIFN-ω reactant mixture for the mPEG propionic aldehyde; M is Marker
The specific embodiment
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol propionic aldehyde (mPEG-propionaldehyde that contracts in recombinant human interferon omega and 1: 1 ratio of Polyethylene Glycol propionic aldehyde mol ratio, mPEG-ALD), stirring and dissolving, add NaCNBH3 again, room temperature reaction 48 hours.
Embodiment 2
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol propionic aldehyde (mPEG-propionaldehyde that contracts in recombinant human interferon omega and 1: 3 ratio of Polyethylene Glycol propionic aldehyde mol ratio, mPEG-ALD), stirring and dissolving, add NaCNBH3 again, room temperature reaction 24 hours.
Embodiment 3
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol propionic aldehyde (mPEG-propionaldehyde that contracts in recombinant human interferon omega and 1: 5 ratio of Polyethylene Glycol propionic aldehyde mol ratio, mPEG-ALD), stirring and dissolving, add NaCNBH3 again, room temperature reaction 12 hours.
Embodiment 4
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol butyral in recombinant human interferon omega and 1: 1 ratio of Polyethylene Glycol butyraldehyde mol ratio, stirring and dissolving adds NaCNBH3 again, room temperature reaction 48 hours.
Embodiment 5
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol butyral in recombinant human interferon omega and 1: 3 ratio of Polyethylene Glycol butyraldehyde mol ratio, stirring and dissolving adds NaCNBH3 again, room temperature reaction 24 hours.
Embodiment 6
The purity of the recombinant human interferon omega of Shi Yonging is greater than 98% in the present embodiment, and maintains higher biological activity; Recombinant human interferon alpha 2 is dissolved in acetic acid-sodium-acetate buffer (pH4.0) of 100mM, add the mono methoxy polyethylene glycol butyral in recombinant human interferon omega and 1: 5 ratio of Polyethylene Glycol butyraldehyde mol ratio, stirring and dissolving adds NaCNBH3 again, room temperature reaction 12 hours.
Embodiment 7
(mobile phase is for containing 0.15M NaCl phosphate buffer pH=7.0 through Superdex 200 gel filtration chromatographies for the product that the reaction of above-mentioned recombinant human interferon alpha 2 and Polyethylene Glycol is obtained, flow velocity 1ml/min) separation and purification, collect each component peaks, after desalination and concentration, be used for the detection of active and purity.
Embodiment 8
Recombinant human interferon alpha 2 and Polyethylene Glycol are reacted the product process cation-exchange chromatography SP-FF separation and purification that obtains, with buffer A (20mM sodium acetate, pH=4.0; Flow velocity 1ml/min) sample on the balance, buffer B (20mM sodium acetate+1M sodium chloride pH=4) linear elution is collected each component peaks, is used for the detection of active and purity after desalination and concentration.
Claims (11)
1. the interferon conjugate of a chemical modification, its structure is shown in the I formula:
R-L-M-NH-(rhIFN-ω) I
Wherein, R represents to have C1 ~ water-soluble polymer of 4 alkane end groups,
L represents O, N, NH atom,
M represents C1 ~ 6 aliphatic saturated hydrocarbon subunits,
NH represents the N-terminal amino of rhIFN-ω,
RhIFN-ω represents human recombinant interferon ω.
2. according to the described conjugate of claim 1, wherein said water-soluble polymer comprises Polyethylene Glycol, polypropylene glycol, polylactic acid, polymeric amino acid, branch's Polyethylene Glycol, branch's polypropylene glycol, branching polymerization aminoacid etc.
3. according to the described conjugate of claim 2, wherein said water-soluble polymer is a Polyethylene Glycol, has the molecular weight between 5kDa ~ 100kDa.
4. according to the described arbitrary conjugate of claim 1 ~ 3, the end group of wherein said water-soluble polymer is a methyl.
5. according to the described conjugate of claim 4, wherein said L represents O or NH.
6. according to the described conjugate of claim 5, wherein said M represents C3 ~ 4 aliphatic saturated hydrocarbon subunits.
7. according to the described conjugate of claim 5, wherein said M represents the straight chain propylidene.
8. the preparation method according to the described arbitrary conjugate of claim 5 ~ 7 is,
(1) provides the buffer system of the salinity of pH4.0 ~ 9.0,10 ~ 100mmol/L;
(2) molar ratio of recombinant human interferon omega and polyethylene glycol aldehyde radical derivative reaction is 0.25 ~ 20;
(3) polyethylene glycol aldehyde radical derivative and recombinant human interferon omega reacted 1 ~ 48 hour in the system that (1) step provides, and temperature is controlled at 4 ℃ ~ 37 ℃;
(4) product that obtains through step (3) reaction is collected the recombinant human interferon omega molecule (rhIFN-ω) of a peg molecule of coupling through sieve chromatography, ion-exchange chromatography, hydrophobic chromatography separation and purification, through filtration sterilization, after the lyophilization promptly.
9. the preparation method of described conjugate according to Claim 8, wherein reaction condition is:
(1) pH value is between 4.5 ~ 6.5, the buffer system of the salinity of 50 ~ 80mmol/L;
(2) molar ratio of recombinant human interferon omega and polyethylene glycol aldehyde radical derivative reaction is controlled between 1 ~ 10;
(3) temperature is controlled at 4 ℃ ~ 25 ℃;
(4) response time is 8 ~ 24 hours.
10. it is characterized in that according to Claim 8 or 9 described Pegylation recombinant human interferon omega (rhIFN-ω) medicine production technologies: Polyethylene Glycol and recombinant human interferon omega (rhIFN-ω) take place the covalent bond reaction needed with sodium cyanoborohydride as catalyst.
11. a pharmaceutical composition that can be used for clinical treatment contains arbitrary conjugate and medicinal diluent, adjuvant or the carrier in the claim 1 ~ 10 of being selected from of effective dose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008103014034A CN101491682A (en) | 2008-04-30 | 2008-04-30 | PEG-IFN omega conjugate and preparation technique thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008103014034A CN101491682A (en) | 2008-04-30 | 2008-04-30 | PEG-IFN omega conjugate and preparation technique thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101491682A true CN101491682A (en) | 2009-07-29 |
Family
ID=40922583
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008103014034A Pending CN101491682A (en) | 2008-04-30 | 2008-04-30 | PEG-IFN omega conjugate and preparation technique thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101491682A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8143214B2 (en) | 2007-08-16 | 2012-03-27 | Pharmaessentia Corp. | Protein-polymer conjugates |
| CN102617736A (en) * | 2010-07-20 | 2012-08-01 | “拜奥卡德”股份公司 | The new stable polyethylene glycol conjugate of interferon alpha, represented by one positional isomer |
| CN105330746A (en) * | 2014-08-14 | 2016-02-17 | 广东紫金正天药业有限公司 | Polyethylene glycol-modified human interferon and human antibacterial peptide fusion protein |
| CN105777892A (en) * | 2014-12-17 | 2016-07-20 | 深圳翰宇药业股份有限公司 | Pramlintide polyethylene glycol derivative, and preparation method and application thereof |
| US11559567B2 (en) | 2014-11-06 | 2023-01-24 | Pharmaessentia Corporation | Dosage regimen for pegylated interferon |
-
2008
- 2008-04-30 CN CNA2008103014034A patent/CN101491682A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8143214B2 (en) | 2007-08-16 | 2012-03-27 | Pharmaessentia Corp. | Protein-polymer conjugates |
| CN102617736A (en) * | 2010-07-20 | 2012-08-01 | “拜奥卡德”股份公司 | The new stable polyethylene glycol conjugate of interferon alpha, represented by one positional isomer |
| CN102617736B (en) * | 2010-07-20 | 2015-11-25 | “拜奥卡德”股份公司 | The stable interferon alpha polyoxyethylene glycol conjugate represented by a kind of positional isomers |
| CN105330746A (en) * | 2014-08-14 | 2016-02-17 | 广东紫金正天药业有限公司 | Polyethylene glycol-modified human interferon and human antibacterial peptide fusion protein |
| US11559567B2 (en) | 2014-11-06 | 2023-01-24 | Pharmaessentia Corporation | Dosage regimen for pegylated interferon |
| CN105777892A (en) * | 2014-12-17 | 2016-07-20 | 深圳翰宇药业股份有限公司 | Pramlintide polyethylene glycol derivative, and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2312976C (en) | Compositions comprising histidine-linked protein polymer conjugates | |
| EP0862455B1 (en) | Interferon-polymer conjugates and process for preparing the same | |
| US5981709A (en) | α-interferon-polymer-conjugates having enhanced biological activity and methods of preparing the same | |
| AU739359B2 (en) | Improved interferon polymer conjugates | |
| CN1088721C (en) | Interferon conjugates | |
| KR101502645B1 (en) | Polyethylene glycol modified interferon alpha 2b and preparation method and applications thereof | |
| CN102145178B (en) | PEG interleukin 15 | |
| KR100694994B1 (en) | Human Granulocyte-Colony Stimulating Factor Isoforms | |
| CN102153643B (en) | Mutant of granulocyte-colony stimulating factor(G-CSF) and chemically conjugated polypeptide thereof | |
| CN101491682A (en) | PEG-IFN omega conjugate and preparation technique thereof | |
| KR101483814B1 (en) | Interferon alpha 2a modified by polyethylene glycol, its synthesis process and application | |
| KR100521309B1 (en) | Glycosylated Human Granulocyte Colony Stimulating Factor(G-CSF) | |
| TW201002350A (en) | Polyethylenenized erythropoietin conjugate, preparation thereof and its use | |
| KR20040020817A (en) | Glycosylated human interferon alpha isoform | |
| KR20040083268A (en) | Human granulocyte-colony stimulating factor conjugate having enhanced stability in blood and process for the preparation thereof | |
| KR100353392B1 (en) | Method of Preparing the PEG-Protein Composition Having Enhanced Biological Activity | |
| KR20090103209A (en) | Human Interleukin-2 Isoforms | |
| KR100473659B1 (en) | Process for preparing hybrid protein replaced by available polymer at the saccharide chain by site-specific modification | |
| CN112175089A (en) | Granulocyte colony stimulating factor recombinant protein, conjugate and application thereof | |
| CN1765926A (en) | Chemically modified interleukin coupler and its preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20090729 |