CN104936610A - Purification method of GLP-1 analogue - Google Patents
Purification method of GLP-1 analogue Download PDFInfo
- Publication number
- CN104936610A CN104936610A CN201280077013.8A CN201280077013A CN104936610A CN 104936610 A CN104936610 A CN 104936610A CN 201280077013 A CN201280077013 A CN 201280077013A CN 104936610 A CN104936610 A CN 104936610A
- Authority
- CN
- China
- Prior art keywords
- glp
- eluent
- acetic acid
- aib
- foregoing
- 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
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
- B01D15/361—Ion-exchange
- B01D15/362—Cation-exchange
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/32—Bonded phase chromatography
- B01D15/325—Reversed phase
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
- B01D15/361—Ion-exchange
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/605—Glucagons
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/26—Glucagons
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Endocrinology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a purification process of (Aib8,35)GLP-1(7-36)NH2 comprising an ion-exchange chromatography purification step and a reversed phase HPLC purification step. The purification process results in high yield of a highly pure GLP-1 analog peptide.
Description
The present invention relates to the purification process of a human glucagon-like-peptide GLP-1, it is characterized in that the method can purification GLP-1 analog (Aib
8,35) GLP-1 (7-36) NH
2, its aminoacid sequence is: His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-IIe-Ala-Trp-Leu-Val-Lys-Aib-Arg-NH
2
GLP-1 analog can synthesize by a kind of mixed method, and described mixed method comprises solid-phase peptide synthesis (SPPS) and liquid phase fragments of peptides coupling method.
Polypeptide (Aib
8,35) GLP-1 (7-36) NH
2refer to mankind's natural polypeptides GLP-1 (7-36) NH
2the derivant of analog, it substituted for the residue of native peptides the 8th (Ala) and the 35th (Gly) with α-aminoacid (Aib).
This polypeptide, its treatment use and its solid-phase peptide synthesis (SPPS) described by disclosed PCT patent application WO2000/34331, but the preparation method described by this patent and be not suitable for large-scale commercial applications produce.
The PCT patent application WO2007/147816 announced describes a kind of synthetic method preparing this GLP-1 analog, and the method utilizes three peptide intermediate fragments to be coupled in the liquid phase to prepare.In addition, another PCT patent application WO2010/033254 then describes a kind of Fmoc solid-state chemical reaction method method and progressively prepares this polypeptide.But no matter using wherein any preparation method, purity is generally less than 95%.
Liquid chromatography people glicentin GLP-1 is used to be widely described in this area.
Such as, according to the description of the PCT patent application WO2007/147816 announced, two these GLP-1 analog of step reversed phase liquid chromatography (RP-HPLC) purification are used.The method employs oxolane as mobile phase, but oxolane eluent is unfavorable for using in extensive reversed phase high-performance liquid chromatography is produced, because it can form peroxide.
First the purification process that PCT patent application WO2011/161007 proposes adopts acid RP-HPLC mobile phase in reversed phase high-performance liquid chromatography, adopts the mobile phase of high ph-values subsequently.But under high ph-values condition, polypeptide is unstable, produces new impurity, between causing batch, purity is inconsistent.
Therefore, be necessary that exploitation commercial scale can obtain high-purity (Aib
8,35) GLP-1 (7-36) NH
2purification process.Be intended to reduce impurity content and corresponding side effect, to increase the toleration for the treatment of.
Applicant finds, by adding ion exchange chromatography as purification (Aib
8,35) GLP-1 (7-36) NH
2an orthogonal step, highly purified product can be obtained, be applicable to commercial mass production level.
The feature of the inventive method is to be separated better has the closely-related impurity of similar hydrophobicity, and these impurity are only by using single RP-HPLC method can not obtain effective purification.
In the present invention, the purity requirement of preferred embodiment final products is higher than 99.3%, and single impurity content is no more than 0.3%.Such as, the purity of final products is about 99.3%, is about 99.4%, is about 99.5%, is about 99.6%, is about 99.7%, is about 99.8%, is about 99.9%, and single impurity content is about 0.3%, is about 0.2%, is about 0.1%.In a further preferred embodiment, the purity of final products is at least 99.9%, and single impurity is less than 0.1%.Highly purified (Aib
8,35) GLP-1 (7-36) NH
2most important in clinical evaluation.
In addition, the load capacity due to ion exchange column is approximately 3 times of reversed phase high-performance liquid chromatography, thus drastically reduce the area purification time.Adopt a kind of preferred embodiment of the present invention, the content of acetate is not more than 0.3%.
Different from the method described by PCT patent application WO 2007/147816 and WO 2011/61007, another advantage of ion-exchange process is the acetonitrile liquid that the first step of purification does not use price higher, can reduce costs further like this.
Finally, the purification process of the present invention product yield that can bring.Such as, purification process of the present invention reaches the yield of at least 15%, the yield of at least 16%, the yield of at least 17%, the yield of at least 18%, the yield of at least 19%, and the yield of at least 20%.Adopt the preferred embodiments of the invention, the yield of about 21% can be reached.
Of the present inventionly theme as polypeptide (Aib
8,35) GLP-1 (7-36) NH
2a kind of purification process, mainly comprise following two steps:
A) ion exchange chromatography step, and
B) reversed phase high-performance liquid chromatography purification step.
In following text, unless otherwise showing, a series of values within the scope of certain comprise its limit value, especially when expression " scope is from certain value ".
Acidic buffer is a kind of containing buffer agent acid solution, can prevent the change of pH value.
" single impurity " refers to non-(Aib
8,35) GLP-1 (7-36) NH
2a peptide species.
" RP-HPLC " refers to reversed phase high-performance liquid chromatography.
Unless otherwise showing, the implication of the various terms that the present invention uses and scope are described below.
For the present invention, " orthogonal " term represents use two complementations but multi-form chromatography carrys out purifying compounds.
According to the present invention, orthogonal chromatographic purification process is made up of two steps:
A) ion-exchange purification method, this is the chromatography based on separated component charge differences, and
B) reversed phase high-performance liquid chromatography, this is based on the different chromatography of component hydrophobicity.
In preferred scheme, a) step performed before b), but also can carry out simultaneously.
According to the present invention, step is a) carry out in acidic buffer.
Step mobile phase a) can be an acidic buffer or its mixture.Therefore, the acidic buffer as step mobile phase a) preferably includes two eluents, eluent A and eluent B, and eluent A and eluent B summation are 100%.Eluent A and B can select from acidic buffer well known to those skilled in the art.Such as, the acidic buffer of acetic acid or its salt, especially its ammonium salt and sodium salt; The acidic buffer of formic acid or its salt, especially its ammonium salt and sodium salt; The acidic buffer of phosphoric acid or its salt, especially its ammonium salt and sodium salt.
Preferred eluent A and B is acetic acid and corresponding a certain salt thereof.Preferred eluent A and B is acetic acid and its corresponding ammonium acetate.
Another scheme of the present invention is, the pH value of step a) acidic buffer should from 1.0 to 6.0 scopes, or in 1.25 to 5.0 scopes, preferred pH value is in 1.5 to 4.6 scopes.
Another scheme of the present invention is, step a) is carried out with two kinds of eluents, i.e. eluent A and eluent B.Preferred version be step a) with ammonium acetate solution as eluent A and acetic acid solution as eluent B.More preferably scheme is, step a) will in step b) before perform, and select ammonium acetate solution and acetic acid solution respectively as step eluent A a) and eluent B.
Preferred version of the present invention is, selection ammonium acetate solution and acetic acid solution are respectively as step eluent A a) and eluent B, and pH value range is from 1.0 to 6.0,1.25 to 5.0, and preferred pH value range is from 1.5 to 4.6.
The present invention more preferably scheme is, step a) acetic acid content gradient increase in ammonium acetate solution mobile phase.
Preferred version is, step a) selects acetic acid solution as eluent B, and its Concentraton gradient scope should in 0 to 60% (v/v), select ammonium acetate solution as eluent A and concentration range from 100 to 40% (v/v).More preferably scheme is, step a) selects acetic acid solution as eluent B, and its Concentraton gradient scope is from 0 to 50% (v/v), and select ammonium acetate solution as eluent A, Concentraton gradient scope is from 100 to 50% (v/v).
An important aspect of the present invention is, ammonium acetate solution is selected in step enforcement a), and pH value range is from 4.0 to 5.0,4.1 to 4.9,4.2 to 4.8,4.3 to 4.7 or 4.4 to 4.6 etc.More preferably scheme is that step a) selects ammonium acetate solution, and pH value range is from 4.2 to 4.8, and preferred pH value range is from 4.4 to 4.5.
According to the present invention, step a) in select ammonium acetate solution, its concentration range is from 0.1 to 50mM, and 0.25 to 40mM, 0.5 to 30mM, or 0.75 arrives 25mM etc.More preferably scheme is, step a) in select ammonium acetate solution, its concentration range from 1 to 20mM, more preferably from 1 to 12mM, further preferred concentration range be 10mM (10 ± 0.5mM) left and right.
An important aspect of the present invention is, step a) selects acetic acid solution, and its pH value range is from 0.5 to 3.0,0.75 to 2.75,0.8 to 2.5,0.9 to 2.25 etc.More preferably scheme be acetic acid solution pH value range from 1.0 to 2.0, more preferably the scope of scheme pH is from 1.5 to 1.7 further.
For step a) acetic acid solution can be dilute solution of acetic acid, especially dilute with water as HPLC grade.
In preferred version, step a) selects acetic acid aqueous solution, and its concentration range is from 40% to 60%, and more preferably scope is 45% to 55%, and more preferably scope is 49 to 51% further.
According to the present invention, step b) adopt reversed-phase high-performance liquid chromatography gradient elution method.
In preferred version, step b) select acidic buffer.In preferred version, step b) select acidic buffer pH value range from 1.0 to 6.0,1.25 to 5.75,1.5 to 5.5, or 1.75 to 5.25 etc.More preferably scope be pH value from 2.0 to 4.5, more preferably scope is that pH value range is from 2.5 to 3 further.
According to the present invention, in step b) in, gradient elution can use one or more acidic effluent liquid, particularly uses two kinds of acidic effluent liquid, i.e. eluent C and eluent D, and two kinds of eluent C and eluent D summation are 100%.Eluent C and eluent D can be a kind of acidic aqueous solution or organic solution.Solution can be the organic solution as acetic acid aqueous solution or acetic acid.Organic solvent is as acetonitrile or ethanol, and wherein ethanol comprises methanol, ethanol and propanol.
In preferred version, step b) select acidic effluent liquid C and acidic effluent liquid D gradient elution, more preferably in scheme, select acetic acid aqueous solution as eluent C and acetic acid acetonitrile solution as eluent D.
Advantage of the present invention is, step b) select gradient from the acidic effluent liquid C of the acidic effluent liquid D and 100 to 30% (v/v) of 0 to 70% (v/v).More preferably scheme is, step b) select gradient from the acidic effluent liquid C of the acidic effluent liquid D and 100 to 40% (v/v) of 0 to 60% (v/v).
Advantage of the present invention is, step b) select acetic acid acetonitrile solution as acidic effluent liquid D, its acetonitrile concentration gradient from 0 to 70% (v/v), and selects acetic acid aqueous solution as acidic effluent liquid C, wherein water concentration gradients 100 to 30% (v/v).More preferably scheme is, step b) select acetonitrile concentration gradient from the acetic acid acetonitrile solution of 0 to 60% (v/v) as the acetic acid aqueous solution of acidic effluent liquid D and water concentration gradients 100 to 40% (v/v) as acidic effluent liquid C.
In another preferred embodiment, step b) when selecting eluent C and eluent D as mobile phase, acetic acid concentration scope is from 0.1 to 0.5N.More in preferred version, step b) select eluent C and eluent D as acetic acid concentration scope during mobile phase from 0.2 to 0.3N.
Advantage of the present invention is, the GLP-1 analog (Aib obtained by purification process of the present invention
8,35) GLP-1 (7-36) NH
2product acetic acid content is less than 10% (w/w).
In a preferred approach, the GLP-1 analog (Aib for preparing of purification process of the present invention
8,35) GLP-1 (7-36) NH
2acetic acid content is no more than 7% (w/w), and in preferred scheme, acetic acid content is no more than 6.5% (w/w).In other preferred version, acetic acid content is less than 6% (w/w).Illustrate with example, (Aib prepared by the inventive method
8,35) GLP-1 (7-36) NH
2containing 6% ± 1% (w/w) acetic acid, 5% ± 1% (w/w) acetic acid, 4% ± 1% (w/w) acetic acid, 3% ± 1% (w/w) acetic acid, 2% ± 1% (w/w) acetic acid, 1% ± 0.5% (w/w) acetic acid, or 0.5% ± 0.1% (w/w) acetic acid.
Spent ion exchange resin can carry out ion exchange fast as immobile phase.Suitable Ion exchange resins type can be chosen from weak cationic resin.Weak cationic resin can from PolyCAT
1000-5, Agilent
wCX, or
choose in WCX-10 resin.More suitably resin is PolyCAT
r resin.
Select silica gel absorber as immobile phase, RP-HPLC can be made to be carried out fast.Suitable silica gel type can be chosen from these silica gel absorbers following:
100-16-C18,
100-10-C18,
100-16-C8,
100-16-C4.
100-10-Phenyl,
eternity-5-C18,
100-5-C4,
c18SMB 100-15HE,
c8SMB 100-15HE,
c4SMB 100-15HE,
sP-120-15-ODS-AP,
sP-120-10-C4-Bio,
sP-200-10-C4-Bio,
100C18,
100C8,
100C4,
sT 150-10-Cl8,
100-10-C18, Gemini110-10-C18, YMC-Triart 120-5-C18 and YMC-Triart 200-10-C8.In above-mentioned silica gel absorber, select
silica gel absorber is especially suitable.
In addition, RP-HPLC is undertaken by selecting the immobile phase based on polymer.Polymer can from PLRP-S 100-10 or Amberchrom
(TM)choose in Profile XT20.
According to the present invention, the purity of the polypeptide products of purification is more than 99%.Such as, the purity of final products is approximately 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%.In preferred embodiment, the purity of polypeptide products more than 99.3%, more preferably over 99.4%.In addition in preferred examples of implementation, the purity of polypeptide products has exceeded 99.5%.
In the present invention, the preferred scheme of another one is, single impurity content is not more than 0.5%, and such as, single impurity content is about 0.3%, 0.2%, or 0.1%.In preferred scheme, single impurity content is no more than 0.3%.
Advantage of the present invention is, step a) and step b) the polypeptide total recovery of two-step purifying can higher than 15%, 16%, 17%, 18%, 19%, or 20%.In preferred scheme, polypeptide total recovery is more than 21%.
In order to obtain the final dry product being applicable to useful in preparing drug formulations, the polypeptide solution can prepared with the purification process of foregoing description is achieved by precipitation or lyophilizing or spray drying technology.
Following examples is only used for carrying out exemplary illustration to the present invention, does not represent restriction the present invention.
test portion
the preparation of embodiment 1:GLP-1 compound
According to the preparation method described by patent application WO 2007/147816 and WO 2009/074483, polypeptide (Aib
8,35) GLP-1 (7-36) NH
23 peptide intermediate fragments by first preparing 3 peptide intermediate fragments, then can be of coupled connections and preparing by crude product in the solution.
The chromatogram purification of product, under the condition of pH 2.75, first carries out ion exchange methods, then carries out purification by reversed phase high-performance liquid chromatography.
embodiment 2: purification process
A) ion exchange chromatography
Ion exchange chromatography method parameter is in table 1.
Table 1
By 100 milligrams of (Aib
8,35) GLP-1 (7-36) NH
2polypeptide dissolving crude product is in the ammonium acetate solution of 4.42 in 10mM pH value, and concentration is 6.67mg/mL, then sample is loaded onto weak cation exchange post.
Due to the protonation to weak-acid ion exchange resin, mobile phase increases the polypeptide of the positively charged that eluting is retained by ion exchange column because of acetic acid concentration gradient contained by it.
Above-mentioned ion exchange chromatography relevant parameter and purge process are respectively in table 2 and table 3
Table 2
Table 3
After fraction collection, the fraction of purity more than 95% carries out merging and lyophilizing.
Freeze-drying prods purity is about 97%, and product yield is about 38%.
The fraction merged uses reversed phase high-performance liquid chromatography purification further.
B) step 2, reversed phase high-performance liquid chromatography (RP-HPLC)
Reversed phase high-performance liquid chromatography purification process parameter is in table 4
Table 4
Ion exchange chromatography is merged the fraction collected to be filled on HPLC purification column, then carries out purification to product reversed phase high-performance liquid chromatography gradient elution.
Above-mentioned reversed phase high-performance liquid chromatography method and parameter are respectively in table 5 and table 6
Table 5
Parameter | Describe |
Eluent C | 0.25N acetic acid aqueous solution |
Eluent D | 0.25N acetic acid acetonitrile solution |
Table 6
Collect fraction, the fraction of purity on 99.3% merges then lyophilizing.
The fraction not reaching purity requirement merges, and then solvent removed by evaporation at reduced pressure uses step b) same condition purification again.
RP-HPLC purification step yield is 70%.Ion-exchange purification and the anti-phase purification total recovery of RP-HPLC are 21%.
GLP-1 analog (the Aib obtained thus
8,35) GLP-1 (7-36) NH
2purity is 99.9%.In purification process, such polypeptide products purity represents high level.
In addition, the GLP-1 analog (Aib obtained thus
8,35) GLP-1 (7-36) NH
2the acetic acid content of polypeptide products is less than 7% (w/w).
Consider the purification process that the present invention announces and index of correlation, other schemes and apply apparent to one skilled in the art.The document that the present invention quotes, comprises patent, patent application and patent open, specifically and intactly introduces text of the present invention at this.The example that the present invention announces and index are at this only as carrying out exemplary illustration, and scope of the present invention and implication are illustrated by following claims.
Claims (19)
1. (Aib
8,35) GLP-1 (7-36) NH
2purification process, comprise two steps:
A) ion exchange chromatography step;
B) reversed phase high-performance liquid chromatography (HPLC) purification step.
2. method according to claim 1, wherein step is a) in step b) carry out before.
3. method according to claim 1, wherein step a) is carried out in acidic buffer.
4. method according to claim 3, wherein step a) is carried out the acidic buffer of pH value range from 1.0 to 6.0, more preferably carries out the acidic buffer of pH value range from 1.5 to 4.6.
5. the method according to any one of foregoing Claims, wherein step a) is carried out as eluent B using ammonium acetate solution as eluent A and acetic acid solution.
6. the method according to any one of foregoing Claims, wherein step a) is carried out with acetic acid content gradient increase in ammonium acetate mobile phase.
7. method according to claim 6, wherein step gradient a) is the ammonium acetate solution of the acetic acid solution as 0% to 60% (v/v) of eluent B and 100% to 40% (v/v) as eluent A, more preferably, as the ammonium acetate solution of the acetic acid solution of 0% to 50% (v/v) of eluent B and 100% to 50% (v/v) as eluent A.
8. the method according to any one of foregoing Claims, wherein step b) correspond to the reverse-phase chromatography adopting gradient elution.
9. method according to claim 8, wherein step b) implement with acidic buffer.
10. method according to claim 9, wherein step b) adopt the acidic buffer of pH value range from 1.0 to 6.0 and more preferably, adopt the acidic buffer of pH value range from 2.0 to 4.5 to implement.
11. method according to claim 8, wherein step b) gradient elution implement as eluent D using acetic acid aqueous solution as eluent C and acetic acid acetonitrile solution.
12. methods according to claim 11, wherein step b) gradient be as eluent D containing 0% to 70% (v/v) acetonitrile solution of acetic acid, with 100% to 30% (v/v) aqueous solution containing acetic acid as eluent C, more preferably, as eluent D containing 0% to 60% (v/v) acetonitrile solution of acetic acid, with as 100% to 40% (v/v) aqueous solution containing acetic acid of eluent C.
13. methods according to claim 11, be wherein used as step b) in mobile phase eluent C and D in acetic acid concentration be 0.1 to 0.5N, more preferably 0.2 to 0.3N.
14. methods according to any one of foregoing Claims, the wherein GLP-1 analog (Aib of gained
8,35) GLP-1 (7-36) NH
2containing the acetic acid lower than 10% (w/w), more preferably, lower than the acetic acid of 7% (w/w).
15. methods according to any one of foregoing Claims, wherein from the GLP-1 analog (Aib that first and second step obtains
8,35) GLP-1 (7-36) NH
2purity be at least 99%, single impurity is lower than 0.5%.
16. methods according to any one of foregoing Claims, wherein from the GLP-1 analog (Aib that first and second step obtains
8,35) GLP-1 (7-36) NH
2purity be at least 99.3%, single impurity is lower than 0.3%.
17. methods according to any one of foregoing Claims, wherein from the GLP-1 analog (Aib that first and second step obtains
8,35) GLP-1 (7-36) NH
2purity be at least 99.9%, single impurity is lower than 0.1%.
18. methods according to any one of foregoing Claims, wherein from the GLP-1 analog (Aib that first and second step obtains
8,35) GLP-1 (7-36) NH
2productive rate be at least 15%.
19. methods according to any one of foregoing Claims, wherein from the GLP-1 analog (Aib that first and second step obtains
8,35) GLP-1 (7-36) NH
2productive rate be at least 20%.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/US2012/064852 WO2014077802A1 (en) | 2012-11-13 | 2012-11-13 | Purification method of a glp-1 analogue |
Publications (1)
Publication Number | Publication Date |
---|---|
CN104936610A true CN104936610A (en) | 2015-09-23 |
Family
ID=47278526
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201280077013.8A Pending CN104936610A (en) | 2012-11-13 | 2012-11-13 | Purification method of GLP-1 analogue |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN104936610A (en) |
WO (1) | WO2014077802A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109311960A (en) * | 2016-03-23 | 2019-02-05 | 巴切姆股份公司 | The purification process of glucagon-like peptide 1 analog |
CN115505035A (en) * | 2022-08-22 | 2022-12-23 | 南京汉欣医药科技有限公司 | Method for purifying semaglutide intermediate polypeptide |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA3228669A1 (en) * | 2021-08-10 | 2023-02-16 | Ganesh Ramachandran | Purification of liraglutide |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2348044A1 (en) * | 1999-03-15 | 2011-07-27 | Novo Nordisk A/S | Ion exchange chromatography of GLP-1, analogs and derivatives thereof |
CN102223890A (en) * | 2008-09-22 | 2011-10-19 | 益普生制造爱尔兰有限公司 | Process for the synthesis of (aib8,35)hglp-1(7-36)-nh2 |
WO2011161007A1 (en) * | 2010-06-21 | 2011-12-29 | F. Hoffmann-La Roche Ag | Reversed phase hplc purification of a glp-1 analogue |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE375393T1 (en) * | 1998-01-30 | 2007-10-15 | Asubio Pharma Co Ltd | METHOD FOR PRODUCING A PEPTIDE USING AN AUXILIARY PEPTIDE |
CZ295890B6 (en) | 1998-12-07 | 2005-11-16 | Societe De Conseils De Recherches Et D'application | GLP-1 analogues having aminoisobutyric acid in position 8 and D-arginine in position 36, their use and pharmaceutical compositions in which the analogues are comprised |
ES2341588T3 (en) | 2006-06-23 | 2010-06-22 | F.Hoffmann-La Roche Ag | SYNTHESIS OF INSULINOTROPIC PEPTIDES. |
BRPI0820765A2 (en) | 2007-12-11 | 2015-06-16 | Hoffmann La Roche | Methods for producing an insulinotropic peptide and peptides |
-
2012
- 2012-11-13 CN CN201280077013.8A patent/CN104936610A/en active Pending
- 2012-11-13 WO PCT/US2012/064852 patent/WO2014077802A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2348044A1 (en) * | 1999-03-15 | 2011-07-27 | Novo Nordisk A/S | Ion exchange chromatography of GLP-1, analogs and derivatives thereof |
CN102223890A (en) * | 2008-09-22 | 2011-10-19 | 益普生制造爱尔兰有限公司 | Process for the synthesis of (aib8,35)hglp-1(7-36)-nh2 |
WO2011161007A1 (en) * | 2010-06-21 | 2011-12-29 | F. Hoffmann-La Roche Ag | Reversed phase hplc purification of a glp-1 analogue |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109311960A (en) * | 2016-03-23 | 2019-02-05 | 巴切姆股份公司 | The purification process of glucagon-like peptide 1 analog |
CN115505035A (en) * | 2022-08-22 | 2022-12-23 | 南京汉欣医药科技有限公司 | Method for purifying semaglutide intermediate polypeptide |
CN115505035B (en) * | 2022-08-22 | 2023-09-05 | 南京汉欣医药科技有限公司 | Purification method of semaglutin intermediate polypeptide |
Also Published As
Publication number | Publication date |
---|---|
WO2014077802A1 (en) | 2014-05-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6593721B2 (en) | A chromatographic method to isolate and purify high-purity recombinant human serum albumin | |
CN103080128B (en) | Reversed phase HPLC purification of a GLP-1 analogue | |
JP4975895B2 (en) | Insulin isolation by high pressure liquid chromatography. | |
CN110903355A (en) | Preparation method of Tirzepatide | |
CN105622726A (en) | Leuprolide acetate preparing method | |
CN104936610A (en) | Purification method of GLP-1 analogue | |
IL248293A (en) | Purification process of gonadotropin | |
CN101555279B (en) | Urinary follicle stimulating hormone with high purity and preparation method thereof | |
CN103087180A (en) | Glucagon-like peptide 1 (GLP-1) analogues with long-acting effect and application thereof | |
CN106290601B (en) | The detection method of amino acid racemization and diastereoisomer impurity in polypeptide drugs | |
CN113024658B (en) | Method for purifying liraglutide | |
EP2931301B1 (en) | Process for purifying insulin and analogues thereof | |
CN106496302B (en) | Method for purifying protein by ion exchange chromatography | |
CN103992378A (en) | Method for preparing Degarelix acetate | |
Zhang et al. | Protein pre-fractionation with a mixed-bed ion exchange column in 3D LC–MS/MS proteome analysis | |
WO2020114002A1 (en) | Method for purifying polypeptide similar to glp-1 | |
AU2014310255B2 (en) | Purification process for PTH | |
CN105017401B (en) | Purification method of ziconotide | |
CN103467593B (en) | Purification method of thymalfasin | |
CN110240628B (en) | Purification method of hydrophilic short peptide | |
WO2014077801A1 (en) | Purification process for preparing highly pure taspoglutide | |
CN112279895B (en) | Preparation method of chemically synthesized acidic polypeptide | |
CN113683663A (en) | Purification method of organism-protected polypeptide crude product | |
CA2592014C (en) | Purified rhigf-i/rhigfbp-3 complexes and their method of manufacture | |
CA2907521C (en) | Synthesis of cyclic imide containing peptide products |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
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 |
Application publication date: 20150923 |