CN105017408B - Pegylation thrombopoietin mimic peptide homotetramer and application thereof - Google Patents
Pegylation thrombopoietin mimic peptide homotetramer and application thereof Download PDFInfo
- Publication number
- CN105017408B CN105017408B CN201410181486.3A CN201410181486A CN105017408B CN 105017408 B CN105017408 B CN 105017408B CN 201410181486 A CN201410181486 A CN 201410181486A CN 105017408 B CN105017408 B CN 105017408B
- Authority
- CN
- China
- Prior art keywords
- npc
- mimic peptide
- homotetramer
- pegylation
- ala
- 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.)
- Active
Links
Abstract
The preparation method and its usage of the present invention relates to a kind of Pegylation thrombopoietin mimic peptide homotetramer with long-acting.Pegylation thrombopoietin mimic peptide homotetramer made of being coupled by the thrombopoietin mimic peptide dimer that amino terminal is cysteine residues with peg molecule both ends, have the function of long-acting raising peripheral blood platelet counts, it can be used as thrombopenia caused by a kind of therapeutic agent of thrombopenia, including Idiopathic Thrombocytopenic Purpura, chemotherapy, decrease of platelet syndrome and myelodysplastic syndrome etc. caused by hepatitis.
Description
Technical field
The present invention relates to the preparations and purposes of a kind of Pegylation thrombopoietin mimic peptide homotetramer, belong to
Biotechnology and drug field.
Background technique
Thrombopenia is the disease being clinically commonly encountered, and is common in tumor radiotherapy, chemotherapy and Patients Following Bone Marrowtransplantation,
See Idiopathic Thrombocytopenic Purpura (ITP), thrombocytopenic syndromes, the myelodysplastic syndrome that hepatitis induces
(MDS) and chronic liver disease.Thrombopenia is a kind of common blood disease, after tumor patient receives chemotherapy and radiation, marrow
Inhibited by different degrees of, causes anaemia, granulocyte reduction and decrease of platelet.Clinically have the long-acting and short-acting of recombination
EPO and G-CSF treatment anaemia and agranulocytosis, short-acting recombination IL-11 and TPO are used for decrease of platelet after chemicotherapy
Disease, long-acting Nplate are used for the treatment of idiopathic thrombocytopenic purpura.
Thrombopoietin (Thrombopoietin, TPO) is to adjust the most important cell factor of thrombocytopoiesis, its energy
The growth and development of single-minded promotion megacaryocyte and the generation of blood platelet.Human endogenous TPO is made of 332 amino acid, and N-terminal is
Its receptor binding site, C-terminal are high glycosylation part (Bartley et al., Cell 77:1117-1124 (1994);
Chang et al., Journal of Biological Chemistry 270:511-514 (1995)).TPO is mainly in liver
Dirty, marrow stromal cell and kidney synthesis, by the way that in conjunction with its receptor c-Mpl, activation signal access promotes megakaryocyte proliferation
Differentiation to mediating platelet generates (Blood 82:1395-1401,1993；Blood 87:2162-2170,1996).
Thrombopoietin mimic peptide (Thrombopoietin Mimetic Peptide, TMP) is to TPO with similar
Function but the peptide material for not having homologous structure screen high activity from filamentous phage display peptide library by Cwirla etc.
TPO simulating peptide (AF12505).The thrombopoietin mimic peptide is linear peptides, and amino acid group becomes Ile-Glu-Gly-
Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-Ala has stimulation TPO dependent cell strain Ba/ F3-mpl
Expanding capacity, EC50 is 400nmol/L (Science 276:1696-1699,1997).But in the active peptide body of the structure
Biological activity is relatively low, and half-life period is extremely short, it is difficult to meet a kind of needs of the clinical application as curative drug.And further
The study found that TMP will not be as recombination TPO, induction generates the neutralizing antibody (Nature with hTPO cross reaction in vivo
Biotechnology 15:1261-1265,1997), prompt the potential applicability in clinical practice for having better than TPO.
To overcome TMP polypeptide half-life period too short defect in vivo, Amgen company designs and has succeeded in developing recombination
TMP-FC fusion protein does not cut off chronic immunity thrombocytopenic purpura (ITP) for clinical treatment splenectomy and spleen；Tool
There is one week long-acting being subcutaneously administered once, obtains U.S. FDA approval listing, trade name Nplate in August, 2008.
Nplate includes 2 identical single-stranded subunits, and each single-stranded comprising the constant region domains IgG Fc and TPO simulating peptide, C-terminal is by two
Sulfide linkage links, and wherein TPO simulating peptide includes 2 concatenated TMP polypeptide sequences, and internal long-acting function (CN is realized in the region Fc
1810832A).
The present invention provides a kind of dimers of thrombopoietin mimic peptide derived from TPO simulating peptide AF12505, should
Dimer passes through poly- at homologous four containing the sulfydryl and bis-activated peg molecule covalent coupling that amino terminal is cysteine
Body conjugate.
The present inventor it has been investigated that, the Pegylation thrombopoietin mimic peptide homotetramer conjugate realize
The very big extension of internal blood halflife remarkably promotes peripheral blood platelet count after single injection administration in experimental animal body
The effect of amount increases can be used as the drug or pharmaceutical composition of a kind of novel long-acting treatment thrombopenia.
Summary of the invention
The object of the present invention is to provide a kind of homotetramer of thrombopoietin mimic peptide with long-acting, with
And realize the preparation method of the homotetramer, for the drug as treatment thrombopenia.
The amino acid sequence of thrombopoietin mimic peptide dimer (code name NPC) is derived from TPO simulating peptide in the present invention
AF12505 has the feature that
NH2-Cys-(Gly)n-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-
Ala-(Gly)m-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-Ala-COOH
Wherein n and m is the integer of 5-10, and (Gly) n is preferably 5, i.e. GlyGlyGlyGlyGly；(Gly) m is preferably 8, i.e.,
GlyGlyGlyGlyGlyGlyGlyGly。
Furthermore technical staff can be designed the sequence that (Gly) n and (Gly) m is 5-10 Gly by conventional method in that art, and
The realization of this patent is not influenced.
Thrombopoietin mimic peptide dimer sequence SEQ ID NO:1 provided by the invention is noteworthy characterized by N-terminal ammonia
Base acid residue is cysteine.
It is a further object of the present invention to provide the preparation methods and reality that prepare thrombopoietin mimic peptide homotetramer
Apply process.Pass through free sulfhydryl groups (or thio group) In of thrombopoietin mimic peptide dimer amino terminal cysteine
Disulfide bond is formed under certain reaction condition to form.
Another program that the present invention is implemented is by bis-activated peg molecule as conjugate, with thrombocytopoiesis
It is even that the free sulfhydryl groups of element simulation peptide dimer amino terminal cysteine form covalent polyethylene glycol under certain reaction condition
Join homotetramer, has the feature that
Bis-activated peg molecule (PEG) is a kind of methoxyl group or hydroxyl, both-end of not containing by identical group
Activation, the polyethylene glycol that covalent coupling can occur with thio group.Including bis-activated maleimide polyethylene glycol, feature is
mal-PEG-mal；Or including bis-activated mercapto-polyglycol, feature SH-PEG-SH;Or including bis-activated two sulphur pyrrole of neighbour
Pyridine polyethylene glycol, feature OPSS-PEG-OPSS；Or including bis-activated vinyl sulfone polyethylene glycol, feature VS-PEG-
VS。
Peg molecule for thrombopoietin mimic peptide analog homodimer in the present invention can be small point
Son or macromolecular, molecular weight ranges are from 5,000-40,000 dalton.It is greater than 5,000 peg molecule by molecular weight
It can realize as dimer conjugate by the significant extension of thrombopoietin mimic peptide analog dimer half-life period in vivo.
Preferred peg molecule size is between 20,000-40,000 dalton.
It is a further object of the present invention to provide the chemical solid phase conjunctions for being used to prepare thrombopoietin mimic peptide dimer
At or recombinant DNA technology method.Recombinant DNA technology, which refers to, generates plain simulating peptide two using expression vector insertion encoding platelet
After the cDNA genetic fragment of aggressiveness, host cell is converted.Fermented again or culture, purifying prepare the technology mistake of corresponding recombinant polypeptide
Journey.
Another object of the present invention provides separation and prepares thrombopoietin mimic peptide analog homodimer
Method, including commonly known ion-exchange chromatography, affinity chromatography, hydrophobic chromatography and reversed thin layer chromatography, molecular exclusion color
The technological means such as spectrum and ultrafiltration.
Another object of the present invention, outside the raising for providing Pegylation thrombopoietin mimic peptide homotetramer
All blood Platelets can be used as the drug or pharmaceutical composition for the treatment of thrombopenia.
As a kind of pharmaceutical composition, it includes pharmaceutically acceptable carrier or excipient or diluent, and effective
The homotetramer polypeptide polyethylene glycol conjugation object of the present invention of dosage.
" homotetramer " used in the present invention refers to amino acid sequence through covalent bond and forms complete phase
The tetramer that same thrombopoietin mimic peptide dimer is formed in amino terminal covalent bond.
" polyethylene glycol conjugation " used in the present invention, which refers to, to be distinguished by the polyethylene glycol of different size molecular weight at both ends
Covalent and generation the Pegylation homotetramer with the amino terminal of a thrombopoietin mimic peptide dimer.
In the present invention used " expression vector ", refer to containing expressing the Escherichia coli that promoter includes T7, Tac, Trp, lac
Expression vector, or the Yeast expression carrier containing α secretion factor and AOX or GAP expression promoter.
So far the present invention is described in detail, by reference to following Examples, can be had to the present invention apparent
Understanding.
Figure of description explanation:
1 NPC-PEG of attached drawing35KThe SDS-PAGE electrophoresis and RP-HPLC of-NPC analyzes map.Figure A is SDS-PAGE electrophoresis
Figure, wherein swimming lane 1 is albumen Marker, and swimming lane 2 is NPC-PEG35K- NPC, PEG conjugate compare theory because mobility is slack-off
It is worth bigger than normal；Scheming B is that RP-HPLC analyzes map
2 pMAL-C2X-Nplate plasmid construction schematic diagram of attached drawing.
Attached drawing 3 NPC, NPC-PEG35K- NPC and NPC-PEG40KGroup is in the intracorporal promotion peripheral blood blood platelet of normal mouse
Increased effect.
4 NPC-PEG of attached drawing20K- NPC, Nplate and NPC-PEG35KThe thrombopenia mouse that-NPC is induced in carboplatin
Intracorporal promotion peripheral blood blood platelet increasing action.
5 NPC-PEG of attached drawing20K- NPC, Nplate and NPC-PEG35K- NPC is in the intracorporal promotion periphery of ITP model mice
Blood blood platelet increasing action.
6 NPC-PEG of attached drawing35K- NPC beasle dog subcutaneous administration pharmacokinetic trial.
Following examples are not intended to limitation of the present invention only for illustration purpose.
Specific embodiment
The chemical solid phase synthesis of example 1, thrombopoietin mimic peptide dimer
NPC polypeptid acid sequence is as follows:
SEQ ID NO:1(NPC):
Cys-Gly-Gly-Gly-Gly-Gly-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-
Ala-Arg-Ala-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-
Trp-Leu-Ala-Ala-Arg-Ala
When C-terminal is carboxyl, Wang Shuzhi is selected to carry out the chemical solid phase synthesis of two kinds of polypeptides.After synthesis, will
To the polypeptide resin of Side chain protective group cracked, the C-terminal of NPC polypeptide is broken to form carboxyl from resin.
The above synthesis in solid state is carried out on the Peptide synthesizer of ABI company.First by the alanine threaded tree of C-terminal
Rouge, then amino acid one by one is carried out from C-terminal to N-terminal.After to be synthesized, Deprotection and from resin be broken after,
NPC polypeptide is purified with reverse hplc chromatography (Waters company, C18 prepare column).Purification condition is that A phase contains 0.5%(V/
V) the aqueous solution of acetic acid, B phase are containing 80% acetonitrile and 0.5%(V/V) aqueous solution of acetic acid, 0-100%B liquid gradient elution.Collect mesh
Polypeptide, it is freeze-dried at dry powder.Through mass spectral analysis, the molecular weight of NPC polypeptide is 3990.5 dalton.
Example 2, NPC polypeptide being prepared by recombinant in Escherichia coli
According to the amino acid sequence of NPC in example 1, its cDNA sequence is designed using Escherichia coli preference codon, by big
Lian Baosheng biotech firm full genome is artificial synthesized.By the cDNA segment of synthesis (in insertion pMB-19T plasmid), with BamH2 and
It is recycled after III double digestion of Hind, large fragment will be recycled after the same double digestion of recombination, amalgamation and expression pMAL-c2X plasmid.In T4 connection
Under enzyme effect, NPC genetic fragment and pMAL-c2X are ligated and transformed into DH5 α bacterium.The NPC gene containing insertion is selected through flat screen
Recombinant plasmid is named as pMAL-c2X-NPC.CaCl is used again2Method conversion expression host strain BL21(DE3) in, obtain and recombinantly express bacterium
Strain generates the fusion protein of MBP-NPC through 1mMol/LIPTG inducing expression.The fusion protein chromatographs pure through Ni-Sepharose
After change, MPB(myelin basic protein is removed with enterokinase digestion), then NPC polypeptide is recombinated with C18 reverse phase column purification, it is lyophilized into dry
Powder.
The cDNA sequence of SEQ ID NO:2(NPC):
tgtggtggtg gtggtggtat tgaaggtcct actttgagac aatggttggc tgctagagct 60
ggtggtggtg gtggtggtgg tggtattgaa ggtcctactt tgagacaatg gttggctgct 120
agagct 126
Example 3, NPC-PEG20KThe preparation of-NPC homotetramer
Polyethylene glycol (the mal-PEG that the bismaleimide that average molecular weight is 20kD is activated20K- mal) it presses
20.0mg/ml is dissolved in 100mMol/L acetate buffer (pH6.5), under the conditions of being stirred at room temperature, is gradually added into NPC polypeptide to two
The molar concentration of person reaches mal-PEG20K- mal: NPC is 1:2 ~ 4.After reaction 4-12 hours, with RP-HPLC through ELSD (evaporation
Light) detection PEG coupling dimer reach 85% or more.The TFA for being added 1% terminates reaction, is separated off with reversed C18 column purification
Unmodified NPC polypeptide, free mal- PEG20K-- mal and mono-modified object PEG20K-- NPC, NPC- PEG after purification20K--
NPC homotetramer is lyophilized into dry powder preservation.
Example 4, NPC-PEG35KThe preparation of-NPC homotetramer
Polyethylene glycol (the mal-PEG that the bismaleimide that average molecular weight is 35kD is activated35K- mal) it presses
20.0mg/ml is dissolved in 100mMol/L acetate buffer (pH6.5), under the conditions of being stirred at room temperature, is gradually added into NPC polypeptide to two
The molar concentration of person reaches mal-PEG35K- mal: NPC is 1:2 ~ 4.After reaction 4-12 hours, with RP-HPLC through ELSD (evaporation
Light) detection PEG coupling dimer reach 80% or more.The TFA for being added 1% terminates reaction, is separated off with reversed C18 column purification
Unmodified NPC, free mal- PEG35K- mal and mono-modified object PEG35K- NPC, NPC-PEG after purification35K- NPC is homologous
The tetramer is lyophilized into dry powder preservation.
Example 5, NPC-PEG40KThe preparation of homodimer
By the polyethylene glycol (mal-PEG for single maleimide activation that average molecular weight is 40kD40K) press 20.0mg/ml
Be dissolved in 100mMol/L acetate buffer (pH6.5), under the conditions of being stirred at room temperature, be gradually added into NPC polypeptide to both it is mole dense
Degree reaches PEG40K: NPC 1:1.After reaction 4-12 hours, reached with RP-HPLC through ELSD (evaporative light) detection PEG coupling
85% or more.The TFA for being added 1% terminates reaction, and unmodified NPC and free mal- are separated off with reversed C18 column purification
PEG40K, NPC-PEG after purification40KHomodimer is lyophilized into dry powder preservation.
Example 6, polyethylene glycol thrombopoietin mimic peptide homotetramer the blood platelet-increasing drug effect in normal mouse body
Effect
Kunming mice is taken, 20-25 grams of weight, every group 10, is divided into 4 groups.Blank control group is physiological saline group, experimental group
For NPC, NPC-PEG35K- NPC and NPC-PEG40KGroup presses the abdominal cavity 10.0ug/kg single injection respectively.The 3rd day after administration,
6 days, the 9th day and the 13rd day eyeball ischemic 120ul measure peripheral blood platelet counts.Experimental result is shown, with control group phase
Than three experimental group platelet counts are significantly raised, wherein NPC-PEG35K- NPC organizes the 6th day platelet counts and increases 4-5
Times；It NPC group (not polyethyleneglycol modified) platelet counts elevated levels and holds time and is below polyethyleneglycol modified group of (NPC-
PEG35K- NPC and NPC-PEG40KGroup).
Example 7, Pegylation thrombopoietin mimic peptide homotetramer induce decrease of platelet animal in carboplatin
The blood platelet-increasing drug action of model
Kunming mice is taken, 20-25 grams of weight, is divided into 4 groups, every group 10: control group after carboplatin 150mg/kg is injected intraperitoneally
Injecting normal saline, positive controls are Nplate(Amgen production), experimental group NPC-PEG20K- NPC group, NPC-PEG35K-
NPC group presses the abdominal cavity 25.0ug/kg single injection.The 3rd day after administration, the 6th day, the 9th day and the 13rd day eyeball take blood
120ul measures peripheral blood platelet counts.The experimental results showed that three experimental group platelet counts are bright compared with model group
It is aobvious to increase, the 13rd day NPC-PEG20K- NPC group and NPC-PEG35KIt is normal that-NPC organizes platelet recovery.
Example 8, Pegylation thrombopoietin mimic peptide homotetramer are in ITP decrease of platelet animal model
Blood platelet-increasing drug action
Idiopathic Thrombocytopenic Purpura (ITP) model mice is taken, every group 10, is divided into 4 groups.Experimental group is NPC-
PEG20K- NPC group, NPC-PEG35K- NPC group and Nplate(Amgen production) group, press the abdominal cavity 25.0ug/kg single injection.In
The 3rd day, the 6th day, the 9th day and the 13rd day eyeball takes blood 120ul after administration, measures peripheral blood platelet counts.Experimental result table
Bright, three experimental group platelet counts are significantly raised.
Pharmacokinetics in example 9, Pegylation thrombopoietin mimic peptide homotetramer beasle dog body
Take beasle dog 4, male and female each 2.NPC-PEG is given by 100 μ g/kg dose subcutaneous35KAfter-NPC, respectively at 0
Hour, 0.5 hour, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 24 hours, 30 hours, 36 hours, it is 48 small
When, 72 hours, 96 hours, 120 hours, 144 hours, 168 hours, 192 hours blood sampling 0.5ml, be centrifugated to obtain serum, -80
DEG C save.The peripheral blood platelet counts at corresponding time point are accordingly measured simultaneously.Using self-built double fastener heart Elisa kit,
Measurement blood concentration simultaneously draws Drug-time curve.The experimental results showed that NPC-PEG35KThe half-life period of-NPC is 96 hours, in single
After administration, peripheral blood platelet counts are significantly increased, and can be maintained 7 days or more.
SEQUENCE LISTING
<110>Chongqing Pai Jin Biotechnology Co., Ltd
<120>Pegylation thrombopoietin mimic peptide homotetramer and application thereof
<160> 2
<210> SEQ ID NO:1
<211> 42
<212> PRT
<213> Artificial sequence
<400> 1
Cys Gly Gly Gly Gly Gly Ile Glu Gly Pro Thr Leu Arg Gln Trp Leu
1 5 10 15
Ala Ala Arg Ala Gly Gly Gly Gly Gly Gly Gly Gly Ile Glu Gly Pro
20 25 30
Thr Leu Arg Gln Trp Leu Ala Ala Arg Ala
35 40
<210> SEQ ID NO:2
<211> 126
<212> DNA
<213> Artificial sequence
<400> 2
tgtggtggtg gtggtggtat tgaaggtcct actttgagac aatggttggc tgctagagct 60
ggtggtggtg gtggtggtgg tggtattgaa ggtcctactt tgagacaatg gttggctgct 120
agagct
Claims (5)
1. Pegylation thrombopoietin mimic peptide homotetramer, it is characterized in that passing through thrombopoietin mimic peptide two
The cysteine residues of aggressiveness amino terminal are formed with bis-activated polyethylene glycol conjugation, wherein
(1) sequence signature of thrombopoietin mimic peptide dimer (code name NPC) are as follows:
NH2-Cys-(Gly)n-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-Ala-
(Gly)m-Ile-Glu-Gly-Pro-Thr-Leu-Arg-Gln-Trp-Leu-Ala-Ala-Arg-Ala-COOH
Wherein n and m is the integer of 5-10；
(2) structure feature of Pegylation thrombopoietin mimic peptide homotetramer is NPC- (CH2-CH2)X-NPC
Wherein x is the integer of 100-1000, CH2-CH2For ethylene glycol；
(3) polyethylene glycol of the sulfydryl of the amino terminal cysteine of thrombopoietin mimic peptide dimer (NPC) and activation
Molecule covalent coupling.
2. Pegylation thrombopoietin mimic peptide homotetramer according to claim 1, it is characterised in that
The bis-activated molecular weight polyethylene glycol is 10000-40000 dalton.
3. according to claim 1 with Pegylation thrombopoietin mimic peptide homotetramer described in any one of 2,
It is characterized in that the active group of the bis-activated peg molecule is one of the following: the poly- second two of maleimide activation
The polyethylene glycol that alcohol, the polyethylene glycol of vinyl sulfone activation, the polyethylene glycol of iodoacetamide activation, pyridine disulfide activate.
4. Pegylation thrombopoietin mimic peptide homotetramer of any of claims 1-3 can in preparation
As the application promoted in the increased drug of platelet counts or pharmaceutical composition.
5. application according to claim 4, it is characterised in that the drug or pharmaceutical composition are suitable for treatment idiopathic
Thrombopenia caused by thrombocytopenic purpura, chemotherapy, decrease of platelet syndrome and myelosis caused by hepatitis
The thrombopenias such as abnormal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410181486.3A CN105017408B (en) | 2014-04-30 | 2014-04-30 | Pegylation thrombopoietin mimic peptide homotetramer and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410181486.3A CN105017408B (en) | 2014-04-30 | 2014-04-30 | Pegylation thrombopoietin mimic peptide homotetramer and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105017408A CN105017408A (en) | 2015-11-04 |
| CN105017408B true CN105017408B (en) | 2019-11-05 |
Family
ID=54407752
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410181486.3A Active CN105017408B (en) | 2014-04-30 | 2014-04-30 | Pegylation thrombopoietin mimic peptide homotetramer and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105017408B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1235979A (en) * | 1999-04-06 | 1999-11-24 | 华子春 | Design of cell factor or growth factor analogue polymer and its use |
| CN1810832A (en) * | 1998-10-23 | 2006-08-02 | 安姆根有限公司 | Dimeric thrombopoietin peptide mimetics binding to MP1 receptor and having thrombopoietic activity |
| CN101227921A (en) * | 2003-06-06 | 2008-07-23 | 医学免疫公司 | Use of EphA4 and modulator of EphA4 for diagnosis, treatment and prevention of cancer |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101993485B (en) * | 2009-08-20 | 2013-04-17 | 重庆富进生物医药有限公司 | Peptide analog homologous dimer capable of accelerating insulin secretion and application thereof |
-
2014
- 2014-04-30 CN CN201410181486.3A patent/CN105017408B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1810832A (en) * | 1998-10-23 | 2006-08-02 | 安姆根有限公司 | Dimeric thrombopoietin peptide mimetics binding to MP1 receptor and having thrombopoietic activity |
| CN1235979A (en) * | 1999-04-06 | 1999-11-24 | 华子春 | Design of cell factor or growth factor analogue polymer and its use |
| CN101227921A (en) * | 2003-06-06 | 2008-07-23 | 医学免疫公司 | Use of EphA4 and modulator of EphA4 for diagnosis, treatment and prevention of cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105017408A (en) | 2015-11-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101993485B (en) | Peptide analog homologous dimer capable of accelerating insulin secretion and application thereof | |
| KR101651703B1 (en) | FGF21 Mutants and Uses Thereof | |
| TWI621626B (en) | Novel insulin analog and use thereof | |
| CN104271588B (en) | The engineered polypeptide of immunogenicity with enhanced acting duration and reduction | |
| CN103403019A (en) | Engineered polypeptides having enhanced duration of action | |
| KR100694994B1 (en) | Human Granulocyte-Colony Stimulating Factor Isoforms | |
| CN103957926A (en) | Engineered polypeptides having enhanced duration of action and reduced immunogenicity | |
| CN116098989A (en) | FGF21 variants, fusion proteins and uses thereof | |
| WO2011060583A1 (en) | Nonnatural collagen-like protein and use thereof | |
| JP7174149B2 (en) | GLP1-Fc fusion protein and complex thereof | |
| WO2021222150A2 (en) | Interleukin-2 polypeptides and fusion proteins thereof, and their pharmaceutical compositions and therapeutic applications | |
| EP3237441A1 (en) | Methods of improving yield in recombinant protein production | |
| US20110003741A1 (en) | Novel neurturin conjugates for pharmaceutical use | |
| AU2017357327B2 (en) | Pegylated endostatin analogue and application thereof | |
| IL273368B2 (en) | Pharmaceutical constructs with enhanced binding affinity with albumin | |
| CN101585864B (en) | Nitrogen-terminal fixed-point coupling method for colony stimulating factor of column chromatography granulocyte and coupled product | |
| CN101503457B (en) | Column chromatography PEG and interferon analogue protein N-terminal site-directed coupling method and product thereof | |
| CN102131518B (en) | Comprise the biopolymer conjugate of interleukin-11 analog | |
| CN110172103B (en) | GLP-1 analogue-Fc fusion protein, and preparation method and application thereof | |
| CN105017408B (en) | Pegylation thrombopoietin mimic peptide homotetramer and application thereof | |
| CN113166271A (en) | Fusion polypeptide conjugates with extended half-life | |
| CN112225820B (en) | Recombinant human serum albumin-collagen binding domain fusion protein of tumor specific targeting matrix and application thereof | |
| CN101376676B (en) | PEGylated erythropoietin protein long-acting preparation | |
| KR20230106149A (en) | Growth hormone fusion protein, manufacturing method and use thereof | |
| CN115232215A (en) | Fusion protein ABD/Fc/IL-2 and encoding gene, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 400 700 No. 106 Fenghe Road, Beibei District, Chongqing Applicant after: CHONGQING PAIJIN BIOTECHNOLOGY CO., LTD. Address before: 401332 Chongqing District of Shapingba city science and technology innovation service productivity Xiyong three road building 8 floor Applicant before: CHONGQING PAIJIN BIOTECHNOLOGY CO., LTD. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |