CN101514229A - Human interferon alpha derivative and polyethylene glycol modified substance thereof - Google Patents
Human interferon alpha derivative and polyethylene glycol modified substance thereof Download PDFInfo
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- CN101514229A CN101514229A CNA2009101311492A CN200910131149A CN101514229A CN 101514229 A CN101514229 A CN 101514229A CN A2009101311492 A CNA2009101311492 A CN A2009101311492A CN 200910131149 A CN200910131149 A CN 200910131149A CN 101514229 A CN101514229 A CN 101514229A
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Abstract
The invention discloses human interferon alpha derivative and polyethylene glycol modified substance thereof. The human interferon alpha derivative provided by the present invention is formed by connecting 4-7 glycines on amino end of human interferon alpha. The polyethylene glycol modified substance of the human interferon alpha derivative is formed by connecting polyethylene glycol to the amino end of the human interferon alpha derivative through amido bond. The invention also provides a preparation method for the polyethylene glycol modified substance of the human interferon alpha derivative and use thereof in treating and preventing viral infection or tumor medicament. The polyethylene glycol modified substance of the human interferon alpha derivative does not contain non-amino end modified isomer, and has purity of more than 95%, long in vivo retention time, and widely clinic application prospect.
Description
Technical field
The present invention relates to the biological medicine technology field, be specifically related to pegylated products thereof and the preparation and the purposes of human interferon alpha derivative, human interferon alpha derivative.
Background technology
(interferon is the active protein with broad-spectrum antiviral, antiproliferative and immunomodulatory function that exists in the class body IFN) to human interferon, is one of cytokine that is used for the earliest clinical treatment.Human interferon can be divided into three types, i.e. human interferon-alpha, and human interferon-β, human interferon-gamma also can be further divided into some hypotypes according to the aminoacid sequence difference of various IFN.Prove that through a large amount of clinical studyes human interferon-alpha is a kind of important antitumor and antiviral therapy medicine.At present China clinical most popular mainly be recombinant human interferon-alpha-1a, α-2a and α-1b.
Human interferon-alpha is made up of 165 amino acid, and molecular weight is about 19000 dalton.There is more than 20 gene member in the I type family of finder's interferon alpha at present, and wherein most of encode functional protein has about 90% homology each other on nucleotide level.But, no matter be human interferon-alpha-2a, human interferon-alpha-1b and human interferon-alpha-1a, as pharmaceutical grade protein, its antiviral specific activity only is 1 * 10
8IU/mg, and owing to easily produce antigen antibody reaction, poor stability, the many shortcomings of untoward reaction are very limited on clinical treatment when plasma clearance height, heavy dose.
At present, can be by the synthetic on a large scale human recombination protein of engineered means, prepare human interferon derivative or analogue, solved the immunogenicity problem that heterologous protein causes to a great extent, but still can't overcome poor stability, plasma clearance is fast and bioavailability is low shortcoming.
The result that these shortcomings cause is: need frequent injection human interferon just can reach effective plasma treatment concentration, and, all can cause after the per injection Plasma Concentration than great fluctuation process, form " Feng Yigu " effect of drug level, so just increased the risk of medical expense and untoward reaction.
Polyethyleneglycol modified proteinic technology is a kind of new technology that is used to improve pharmacokinetics character in the protein drug body that grows up over past ten years.It is that activated polyglycol molecule (PEG) is bonded on the protein molecule surface, thereby influences proteinic space structure, finally causes the change of the various biochemical properties of protein.
PEG is the amphipathic molecule of the inertia long-chain that is polymerized by the oxygen vinyl monomer.Now existing multiple different PEG can be for utilizing.Its active function gene of the PEG that is activated can be coupled to certain privileged sites (such as amino, sulfydryl or other nucleophiles) of treatment molecule, can effectively control the purity of modified outcome, makes the simpler and also easier evaluation of quality product of technology.
Sequential analysis is found: the 1st N-terminal of the α 1a of human interferon-alpha family, α 2a and α 1b is cysteine residues, its-SH and 29 Cys-disulfide linkage formed between the SH, thereby the peptide molecule that makes human interferon-alpha becomes curling sterie configuration, if the n terminal residue of existing human interferon-alpha is modified with Pegylation PEG, its Pegylation modification rate is very low, only reach 2～5%, and the pegylated products thereof that obtains contains non-aminoterminal modifies isomer, influences the purity of the pegylated products thereof of human interferon-alpha.
Summary of the invention
One of purpose of the present invention is to solve the low problem of human interferon-alpha Pegylation modification rate, a kind of human interferon alpha derivative is provided, described human interferon alpha derivative is to connect 4～7 glycine at the N-terminal of human interferon-alpha, and general formula is Gly (n)-IFN α, and wherein n is 4～7.
Two of purpose of the present invention provides a kind of pegylated products thereof of human interferon alpha derivative, promptly on the aminoterminal amino of above-mentioned human interferon alpha derivative, connect activated polyglycol molecule (PEG) by amido linkage, its general molecular formula is as follows, and wherein n is 4～7:
Polyoxyethylene glycol molecular-weight average of the present invention is 10000～40000 dalton.
Three of purpose of the present invention provides the preparation method of the pegylated products thereof of above-mentioned human interferon alpha derivative.
Four of purpose of the present invention provides human interferon alpha derivative and the purposes of pegylated products thereof in the medicine of preparation treatment or prophylaxis of viral infections or tumor disease thereof.
Technical scheme of the present invention is as follows:
Human interferon alpha derivative of the present invention is to connect the peptide segment of being made up of 4-7 glycine at the N-terminal of human interferon-alpha.This peptide segment has fully exposed aminoterminal dissociating-NH
2, help PEG to aminoterminal pointed decoration.This peptide fragment preferably is made up of 5 glycine.
Human interferon alpha derivative of the present invention comprises human interferon alpha 1 b (IFN-α 1b), α 2a (IFN-α 2a), α 2b (IFN-α 2b) three kinds of proteic derivatives, analogue, biological activity or pharmaceutical activity fragments.
The present invention also provides the nucleotide sequence of the described human interferon alpha derivative of encoding.In the specific embodiment of the present invention, provide a kind of can be in pichia spp the nucleotide sequence of the human interferon alpha derivative of high expression level, its nucleotide sequence is shown in SEQ ID No.2.
The preparation method of the pegylated products thereof of described human interferon alpha derivative provided by the invention comprises:
1) preparation human interferon alpha derivative;
2) polyoxyethylene glycol and human interferon alpha derivative linked reaction;
3) purifying of the pegylated products thereof of human interferon alpha derivative.
In the specific embodiment of the present invention, connect 4～7 glycine through gene recombination technology at the N-terminal of human interferon-alpha, express described human interferon alpha derivative with the pichia yeast expression system that does not need any inductor, the human interferon alpha derivative of acquisition has fully exposed aminoterminal dissociating-NH
2, help the aminoterminal of the right human interferon alpha derivative of PEG and dissociate-NH
2Modify, have higher modification rate and reach 50%.
Utilize PEG that the N-terminal of human interferon alpha derivative of the present invention is carried out the human interferon alpha derivative carbowax modifier that pointed decoration obtains, stability height, good water solubility, antigenicity are low, and do not contain non-aminoterminal and modify isomer, purity is up to more than 95%, retain in the body for up to 9 days, long-acting is obvious, has good potential applicability in clinical practice.
Description of drawings
Fig. 1 is the structure synoptic diagram of pGAP-IFN Yeast expression carrier;
Fig. 2 is that the PCR of positive expression bacterial classification goal gene identifies;
Fig. 3 is the SDS-PAGE electrophorogram of mPEG-Gly (5)-IFN α;
Fig. 4 is the concentration-time curve figure of mPEG-Gly (5)-IFN α in mouse blood;
Fig. 5 is the concentration-time curve figure of mPEG-Gly (5)-IFN α in rat blood;
Fig. 6 is the concentration-time curve figure of fugitive Interferon, rabbit in rat blood.
Embodiment
Describe the present invention by the following examples in detail.Related in the following embodiments experiment material all can buy or pass through preparation method's acquisition of this area routine if no special instructions by market.
Embodiment 1:gly-gly-gly-gly-gly-IFN α-2a is (hereinafter to be referred as Gly (5)-IFN
α) the secreting, expressing in pichia spp
1, the design of goal gene and acquisition:
The aminoacid sequence of IFN α-2a is shown in SEQ ID No.1.Make corresponding codon into the yeast preferences know the CDNA of IFN α-2a by Genebank after, and add the corresponding nucleotide sequence of gly-gly-gly-gly-gly at the N end.This CDNA sequence is used for the structure of the pGAPZ alpha expression plasmid (available from invitrogen) of pichia spp GS115 (available from invitrogen).Realize secreting, expressing after transforming GS115 host bacterium.Therefore add the enzyme recognition site CTC GAG AAA AGA of KEX2 in design, wherein CTC GAG is the XhoI restriction enzyme site.Two terminator codon TGA TAA of 3 ' end introducing simultaneously and NotI enzyme are cut sequence GCG GCCGC.The CDNA sequence of IFN that is used for Yeast expression carrier is shown in SEQ ID No.2.
Described CDNA sequence is synthetic by the Shanghai biotechnology total man worker of company limited gene, and on the pBluescript II SK (+) (available from Fermentas life sciences) that packs into after the order-checking, the insertion site is XhoI/NotI.Called after pblue-IFN.
2, the structure of plasmid pGAPZ α-IFN.
Goal gene among the pblue-IFN is reclaimed the purpose fragment with XhoI and NotI endonuclease double digestion glue, be equipped with connection.
PGAPZ α plasmid is reclaimed big fragment as carrier with endonuclease XhoI and NotI double digestion, be connected with the T4 ligase enzyme with above-mentioned IFN fragment, 16 ℃ are spent the night, and use CaCl again
2Method is transformed among the E.coli DH5 α, uses the enzyme cutting method screening positive clone, called after pGAPZ α-IFN, and the structure synoptic diagram of pGAPZ α-IFN plasmid is as shown in Figure 1.
3, express the acquisition of engineering bacteria
PGAPZ α-IFN plasmid is transferred among the yeast GS115 after with linearization of BspHI, and continues the selection of positive recombinant, its process is:
(1) preparation competence yeast GS115;
(2) the plasmid pGAPZ α-IFN of linearization of adding;
(3) join behind the mixing in the conversion cup;
(4) carry out electricity with electroporation apparatus and transform click conditional: 0.5kv, 2.5Uf, 1500hm;
(5) use 1.0ml, the 1.0M sorbyl alcohol washes out;
(6) coat and contain on the Zeocin flat board, 30 ℃ of cultivations grow single bacterium colony after 24 hours;
(7) picking list bacterium colony recoats and is distributed on the flat board that contains different concns Zeocin, with purifying and screening high expression level bacterium;
(8) with the goal gene in the method evaluation positive expression bacterial classification of PCR, the result shows that the positive expression bacterial classification contains goal gene as shown in Figure 2.
4, the expression of engineering bacteria
Picking mono-clonal engineering bacteria on the flat board of Zeocin is cultivated in the YPD of 10ml nutrient solution, and 30 ℃, 300rpm spends the night.Second day cultivation bacterium with 0.1ml is placed among the YPD of 50ml and cultivates with similarity condition, respectively 0,24,48,72,96 hours results 1ml bacterium liquid is with the method check protein expression of SDS-PAGE, experimental result shows that protein expression appearred in Gly (5)-IFN α at 24 hours, and is the highest at 48 hours expression amounts.
The purifying of embodiment 2:Gly (5)-IFN α
The first step: positively charged ion gel column (CM Sepharose F.F. gel is available from AmershamBiosciences) chromatography:
Adopt pH3.8～4.6 acetate buffers to carry out upper prop, wash-out, target compound is collected in the electrophoresis monitoring.Use pH7.5～8.5 Tris-HCl buffered soln that target compound is dialysed then.
Second step: negatively charged ion gel column (DEAE Sepharose F.F. gel is available from AmershamBiosciences) chromatography:
Adopt pH7.5～8.5 Tris-HCl buffered soln to carry out upper prop, wash-out, collect target compound.With pH7.5～8.5 phosphate buffer solns target compound is dialysed again.
The preparation and the purifying of sample modified in embodiment 3:PEG coupling
1, embodiment 1 gained Gly (5)-IFN α sample is dialysed with phosphate buffered saline buffer (PH is 6.0), the ALD-PEG 40KD (available from the Jiankai Science and Technology Co., Ltd., Beijing) that added mol ratio then and be 1: 3 modifies at 2～15 ℃, and the reaction times is 40 hours.The modification sample that obtains is carried out SDS-PAGE detect, test-results shows that after the PEG coupling was modified, molecular weight improved, and brings up to nearly 90000 dalton by 19000 original dalton, and the modification rate reaches about 60%, obtains target compound mPEG-Gly (5)-IFN α.
2, the purifying of mPEG-Gly (5)-IFN α:
Use acetate buffer to regulate (the pH value is 4.0～5.0) with termination reaction mPEG-Gly (5)-IFN α, Shangyang ionic gel post SP Sepharose F.F. gel column carries out purifying.Carry out wash-out with NaCl (0.15M) solution, collect target compound, purity reaches more than 95%.The result as shown in Figure 3.
Embodiment 4:ELISA method detects mPEG-Gly (5)-IFN α at intravital concentration of mouse and time-varying relationship
Get kunming mice male and female half and half, body weight is about the 22-25 gram, pegylated products thereof mPEG-Gly (the 5)-IFN α of mouse peritoneal injection embodiment 3 gained human interferon alpha derivatives, dosage is 50 microgram/kilograms, eye socket blood sampling in 24,72,120,168,216 hours after administration, every mouse blood sampling 0.8-1.0ml does not add antithrombotics, 4 mouse of at every turn taking a blood sample.Adopted centrifuge tube on the blood bonnet, left standstill 30 minutes, the glass capillary of taking the bacterium of having gone out then gently scrape a figure around the centrifuge tube tube wall, the blood that solidifies is scraped from tube wall.2000 rev/mins centrifugal 10 minutes, get supernatant and put-20 ℃ of refrigerators and preserve, treat that all serum has all been adopted then to detect according to the specification sheets on the ELISA test kit.The concentration of pegylated products thereof in mouse blood through different time human interferon alpha derivative behind the internal metabolism is as shown in table 1, and the change curve of its concentration and time as shown in Figure 4.
The concentration (pg/ml) of table 1mPEG-Gly (5)-IFN α in mouse blood
Experiment conclusion: use the content that the enzyme linked immunoassay method detects mPEG-Gly (5) in the mouse body-IFN α, as can be known behind 50 micrograms/kg mPEG-Gly (5)-IFN α sample intraperitoneal injection of mice, when being 9 days in 216 hours, can both detect, relatively with prior art in the interior retention time of pegylated products thereof 6-7 days body of human interferon alpha derivative, mPEG-Gly (5)-IFN α retention time in vivo obviously prolongs.
Embodiment 5:mPEG-Gly (5)-IFN α and recombinanthumaninterferon are in the intravital Plasma Concentration of rat relatively
Get 9 of cleaning level SD rats, male, body weight is about 200g.Be divided into 2 groups at random, be 2 of 7 of long-acting interferon groups and fugitive Interferon, rabbit groups.Pegylated products thereof mPEG-Gly (the 5)-IFN α dosage of long-acting interferon group abdominal injection embodiment 3 gained human interferon alpha derivatives is 50 microgram/kilograms, 24,72,120,168,216 and 264 hours eye socket blood sampling 0.8ml after administration, 2500 rev/mins centrifugal 10 minutes, get supernatant and put-20 ℃ of refrigerator cold-storages to be measured.Fugitive Interferon, rabbit group abdominal injection streat drug recombinant human interferon-alpha injection liquid (Huaxin Advanced Biotechnical Co., Ltd., Shanghai's production); dosage is 50 microgram/kilograms; at administration 4,10 and 12 hours blood sampling 0.8ml, 2500 rev/mins centrifugal 10 minutes, get supernatant and put-20 ℃ of refrigerator cold-storages to be measured.Treat the whole content that finish with human interferon-a in the enzyme linked immunoassay method survey serum of collecting of all serum.Shown in table 2,3, the change curve of its concentration and time is shown in Fig. 5,6 in the concentration in the mouse blood for the pegylated products thereof of different time human interferon alpha derivative behind the process internal metabolism.
The concentration (pg/ml) of table 2mPEG-Gly (5)-IFN α in rat blood
The concentration (pg/ml) of the fugitive Interferon, rabbit of table 3 in rat blood
Experiment conclusion: use the enzyme linked immunoassay method and detect the Plasma Concentration of Interferon, rabbit rat, dosage is that the fugitive Interferon, rabbit recombinant human interferon-alpha medicine of 50 micrograms/kg is 4 hours for peak value as can be known, and metabolic process is rapid, 10 hours basic baseline values that arrive; And behind the mPEG-Gly of 50 micrograms/kg (5)-IFN α sample abdominal injection rat, when being 9 days in 216 hours, can both detect, illustrate that its metabolism in vivo reaches long lasting effect.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
SEQUENCE?LISTING
＜110〉Hainan Sihuan Cardiovascular ﹠ Cerebrovascular Medicines Research Institute Company Limited
The Beijing Sihuan Pharmaceutical Co., Ltd
Hainan Sihuan Pharmaceutical Co., Ltd
＜120〉human interferon alpha derivative and pegylated products thereof thereof
<130>MP090344
<160>2
<170>PatentIn?version?3.3
<210>1
<211>165
<212>PRT
＜213〉IFN α-2a albumen
<400>1
Cys?Asp?Leu?Pro?Gln?Thr?His?Ser?Leu?Gly?Ser?Arg?Arg?Thr?Leu?Met
1 5 10 15
Leu?Leu?Ala?Gln?Met?Arg?Arg?Ile?Ser?Leu?Phe?Ser?Cys?Leu?Lys?Asp
20 25 30
Arg?His?Asp?Phe?Gly?Phe?Pro?Gln?Glu?Glu?Phe?Gly?Asn?Gln?Phe?Gln
35 40 45
Lys?Ala?Glu?Thr?Ile?Pro?Val?Leu?His?Glu?Met?Ile?Gln?Gln?Ile?Phe
50 55 60
Asn?Leu?Phe?Ser?Thr?Lys?Asp?Ser?Ser?Ala?Ala?Trp?Asp?Glu?Thr?Leu
65 70 75 80
Leu?Asp?Lys?Phe?Tyr?Thr?Glu?Leu?Tyr?Gln?Gln?Leu?Asn?Asp?Leu?Glu
85 90 95
Ala?Cys?Val?Ile?Gln?Gly?Val?Gly?Val?Thr?Glu?Thr?Pro?Leu?Met?Lys
100 105 110
Glu?Asp?Ser?Ile?Leu?Ala?Val?Arg?Lys?Tyr?Phe?Gln?Arg?Ile?Thr?Leu
115 120 125
Tyr?Leu?Lys?Glu?Lys?Lys?Tyr?Ser?Pro?Cys?Ala?Trp?Glu?Val?Val?Arg
130 135 140
Ala?Glu?Ile?Met?Arg?Ser?Phe?Ser?Leu?Ser?Thr?Asn?Leu?Gln?Glu?Ser
145 150 155 160
Leu?Arg?Ser?Lys Glu
165
<210>2
<211>536
<212>DNA
＜213〉the CDNA sequence of IFN α-2a
<400>2
ctcgagaaaa gaggaggcgg tgggggttgt gatcttccta aaacacactc
cttgggatct 60
aggcgtacat taatgttgtt agcccagatg agacgaatct ctttgttttc
gatagacatg attttggatt cccaaaagag gaattcggta accagtttca
aaaagctgaa 180
accattcccg tgctgcatga gatgattaaa aaaatattca acctattctc
tactaaggac 240
tcatccgcag cttgggacga aaccctgttg gataagttct acaccgaact
ttaccaaaaa 300
ctgaatgacc ttgaggcatg tgttattcag ggcgtcggtg taactgaaac
tccattgatg 360
aaagaggata gtatccttgc tgttaggaag tactttcagc gtattacact
atatctaaaa 420
gaaaaaaagt attctccttg tgcctgggaa gtcgttagag ccgaaatcat
gagatctttt 480
agtttgtcaa?ctaatctgca?ggagagtcta?agatcaaagg?agtaatgagc?ggccgc
536
Claims (10)
1. a human interferon alpha derivative is characterized in that, connects the peptide segment of being made up of 4-7 glycine at the N-terminal of human interferon-alpha.
2. human interferon alpha derivative according to claim 1 is characterized in that, described human interferon-alpha is selected from human interferon alpha 1 b, human interferon-alpha 2a, human interferon-alpha-2 b and described three kinds of proteic derivatives and analogue.
3. human interferon alpha derivative according to claim 1 is characterized in that, connects the peptide segment of being made up of 5 glycine at the N-terminal of human interferon-alpha.
4. the nucleotide sequence of each described human interferon alpha derivative of coding claim 1-3.
5. the expression vector that contains the described nucleotide sequence of claim 4.
6. according to the expression vector of claim 5, it is characterized in that described expression vector is a pichia spp.
7. according to the pegylated products thereof of each described human interferon alpha derivative of claim 1-3, it is characterized in that polyoxyethylene glycol is connected the N-terminal of described human interferon alpha derivative with amido linkage.
8. according to the pegylated products thereof of the described human interferon alpha derivative of claim 7, it is characterized in that the polyoxyethylene glycol molecular-weight average is 10000～40000 dalton.
9. according to the preparation method of the pegylated products thereof of claim 7 or 8 described human interferon alpha derivatives, may further comprise the steps:
1) preparation human interferon alpha derivative;
2) polyoxyethylene glycol and human interferon alpha derivative linked reaction;
3) purifying of the pegylated products thereof of human interferon alpha derivative.
10. according to the application in the medicine of preparation treatment or prophylaxis of viral infections or tumor disease of the carbowax modifier of claim 7 or 8 described human interferon alpha derivatives.
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| CN101514229A true CN101514229A (en) | 2009-08-26 |
| CN101514229B CN101514229B (en) | 2012-05-09 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8143214B2 (en) | 2007-08-16 | 2012-03-27 | Pharmaessentia Corp. | Protein-polymer conjugates |
| CN102453089A (en) * | 2010-10-25 | 2012-05-16 | 重庆富进生物医药有限公司 | Preparation and application of recombinant consensus interferon mutant polyethylene glycol conjugate |
| 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 |
| CN103933577A (en) * | 2010-10-25 | 2014-07-23 | 北京凯因科技股份有限公司 | Preparation and application of recombinant interferon variant polyethylene glycol conjugate |
| US11559567B2 (en) | 2014-11-06 | 2023-01-24 | Pharmaessentia Corporation | Dosage regimen for pegylated interferon |
| WO2024104412A1 (en) * | 2022-11-16 | 2024-05-23 | I-Mab Biopharma Co., Ltd. | Attenuated interferon proteins and fragments and multifunctional polypeptides and conjugates |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2005118752A (en) * | 2002-11-15 | 2006-02-10 | Ф.Хоффманн-Ля Рош Аг (Ch) | POSITIONAL ISOMERS OF PEGLIATED INTERFERON L-2A (PEG-IFN-L-2A CONJUGATE) |
| CN1544468A (en) * | 2003-11-24 | 2004-11-10 | 中国药科大学 | Polyethylene glycol-interferon connected with amido bond and its making method and uses |
-
2009
- 2009-04-03 CN CN2009101311492A patent/CN101514229B/en active Active
Cited By (8)
| 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 |
| CN102453089A (en) * | 2010-10-25 | 2012-05-16 | 重庆富进生物医药有限公司 | Preparation and application of recombinant consensus interferon mutant polyethylene glycol conjugate |
| CN102453089B (en) * | 2010-10-25 | 2014-06-04 | 北京凯因科技股份有限公司 | Preparation and application of recombinant consensus interferon mutant polyethylene glycol conjugate |
| CN103933577A (en) * | 2010-10-25 | 2014-07-23 | 北京凯因科技股份有限公司 | Preparation and application of recombinant interferon variant polyethylene glycol conjugate |
| US11559567B2 (en) | 2014-11-06 | 2023-01-24 | Pharmaessentia Corporation | Dosage regimen for pegylated interferon |
| WO2024104412A1 (en) * | 2022-11-16 | 2024-05-23 | I-Mab Biopharma Co., Ltd. | Attenuated interferon proteins and fragments and multifunctional polypeptides and conjugates |
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| Publication number | Publication date |
|---|---|
| CN101514229B (en) | 2012-05-09 |
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Effective date of registration: 20220318 Address after: 101114 Beijing city Tongzhou District Zhangjiawan Town Qi Shanzhuang East Village Patentee after: BEIJING SIHUAN PHARMACEUTICAL Co.,Ltd. Patentee after: Hainan four ring Pharmaceutical Co., Ltd. Address before: 570216 a06-2, Haikou Free Trade Zone, Hainan Province Patentee before: Hainan Sihuan Cardiovascular Drug Research Institute Co.,Ltd. Patentee before: BEIJING SIHUAN PHARMACEUTICAL Co.,Ltd. Patentee before: HAINAN SIHUAN PHARMACEUTICAL Co.,Ltd. |
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