CN108794618A - A method of purifying Liraglutide - Google Patents
A method of purifying Liraglutide Download PDFInfo
- Publication number
- CN108794618A CN108794618A CN201810663443.7A CN201810663443A CN108794618A CN 108794618 A CN108794618 A CN 108794618A CN 201810663443 A CN201810663443 A CN 201810663443A CN 108794618 A CN108794618 A CN 108794618A
- Authority
- CN
- China
- Prior art keywords
- liraglutide
- purifying
- gly
- solution
- acetonitrile
- 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.)
- Granted
Links
Classifications
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Toxicology (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 invention discloses a kind of methods of purifying Liraglutide, include the following steps：Step 1, Liraglutide crude product pre-process to obtain the thick peptide aqueous solution of Liraglutide；Step 2, the first time HPLC purifying of removal segment impurity；Step 3 removes solvent, obtains Liraglutide first step sample solution；Step 4 removes second of HPLC purifying of the peptide for the more Gly sequences being synthetically produced, the peptide for lacking Gly sequences, epimer；Step 5 removes partial solvent and obtains Liraglutide second step sample solution, adjusts pH value to 6.5, obtains Liraglutide final sample solution.The present invention can remove the impurity such as peptide, peptide, the epimer for lacking Gly sequences of the more Gly sequences being synthetically produced, improve and draw Shandong peptide sample purity and yield.
Description
Technical field
The present invention relates to peptide purification field, especially a kind of purification process of synthesis in solid state Liraglutide.
Background technology
Liraglutide, English name：Liraglutide.
Peptide sequence is：
H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly--
Gln-
Ala-Ala-Lys(Pal-γ-Glu)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-
OH
First long-acting GLP-1 analog that Liraglutide is developed by Novo Nordisk Co., Ltd of Denmark, with people's glicentin sample
(GLP-1) homology of peptide -1 is up to 97%.Liraglutide has reduction blood glucose, promotes pancreatic cell regeneration, mild prolonged gastric emptying
Etc. a variety of effects, application prospect is extensive.
Liraglutide is as hypoglycemic drug of a new generation based on incretin, not only long action time, but also
It has been sufficiently reserved the multinomial physiological activity of natural GLP-1, it can safe and effective hypoglycemic and may be to a variety of cardiovascular risk factors
Protective effect.Liraglutide injection subcutaneous injection is administered, and peak reaching time of blood concentration is 20~14 hours, half-life period 11
~13 hours, daily injection was primary, provides glycemic control for 24 hours, pharmacokinetic properties are not by gender or age effects.Profit
It is absolutely diabetes B therapy field revolutionary character drug to draw Shandong peptide.
Synthesis in solid state Liraglutide will produce " more Gly ", " scarce Gly " and epimer (such as D-His) in synthetic
The impurity of the purity and yield of sample is influenced, low by the isolated Liraglutide sample purity of existing purification technique, yield is low；
The present invention solves this problem.
Invention content
It to solve the deficiencies in the prior art, can the purpose of the present invention is to provide a kind of method of purifying Liraglutide
It removes the peptide for the more Gly sequences being synthetically produced, lack peptide, epimer (such as D-His) impurity of Gly sequences, improve and draw Shandong
Peptide sample purity and yield.
In order to realize that above-mentioned target, the present invention adopt the following technical scheme that：
A method of purifying Liraglutide includes the following steps：
Step 1, Liraglutide crude product pre-process to obtain the thick peptide aqueous solution of Liraglutide；
Step 2, the first time HPLC purifying of removal segment impurity；
Step 3 removes solvent, obtains Liraglutide first step sample solution；
Step 4, remove the peptide for the more Gly sequences being synthetically produced, second of the peptide for lacking Gly sequences, epimer
HPLC is purified；Second of HPLC purifies the impurity removed：D-His1、D-Ser8、D-Ala19、D-Glu21、D-Arg30、
Endo-Gly31、Endo-Gly29、Des-Gly16。
Step 5 removes solvent and obtains Liraglutide second step sample solution, adjusts pH value to 6.5, obtains Liraglutide
Final sample solution.
A kind of method of purifying Liraglutide above-mentioned, step 1, Liraglutide crude product pre-processes to obtain Liraglutide thick
Peptide aqueous solution；Pretreated step includes：
1) Liraglutide crude product is dissolved in acetonitrile solution and obtains the thick peptide aqueous solution of Liraglutide；
2) Liraglutide aqueous solution membrane filtration is taken to remove insoluble granule.It is spare to collect filtrate.
A kind of method of purifying Liraglutide above-mentioned,
Step 2, the first time HPLC purifying of removal segment impurity, segment impurity packet
It includes:H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-
Gln-OH,
H-Ala-Ala-Lys(Pal-γ-Glu)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-
Gly-OH；
First time HPLC purifying specific steps include：Using eight alkyl silane bonded silica gel fillers as stationary phase, with phosphoric acid
For mobile phase A, using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC linear gradient elutions, collects and draws Shandong containing advantageous
The fraction of peptide sample.
A kind of method of purifying Liraglutide above-mentioned,
Mobile phase A uses 0.5% phosphoric acid solution, phosphoric acid solution to add 5ml phosphoric acid by 1000ml water, is uniformly mixed, with three second
Amine adjusts pH value and obtains to 7.0.
A kind of method of purifying Liraglutide above-mentioned, step 3, with rotary evaporator water bath temperature at 30~35 DEG C, very
Reciprocal of duty cycle removes partial acetonitrile below -0.09MPa, obtains Liraglutide first step sample solution, and sample solution is alkalinity.
A kind of method of purifying Liraglutide above-mentioned, step 4 remove the sequence for the more Gly being synthetically produced, lack Gly's
Second of HPLC of sequence and epimer is purified；
Second HPLC purifying the specific steps are：Using HILIC chromatographic column fillers as stationary phase, with trifluoroacetic acid solution with
The mixed liquor of acetonitrile is mobile phase A, and using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC gradient elution, collects and contain
There is the fraction of Liraglutide sample.
A kind of method of purifying Liraglutide above-mentioned, mobile phase A is 0.08% trifluoroacetic acid solution：Acetonitrile=70V:
The mixed liquor of 30V, 0.08% trifluoroacetic acid solution add trifluoroacetic acid 0.8ml to be mixed by 1000ml water.
A kind of method of purifying Liraglutide above-mentioned, step 5, with rotary evaporator water bath temperature at 30~35 DEG C, very
Reciprocal of duty cycle removes partial acetonitrile below -0.09MPa, obtains Liraglutide second step sample solution, Liraglutide second step sample
Solution contains trifluoroacetic acid, and solution is acidity, with ammonium hydroxide tune PH to 6.5.
The invention has the beneficial effects that：
It is combined with eight alkyl silane bonded silica gel fillers with mobile phase phosphoric acid, it is most not that Liraglutide can be removed
Know impurity and segment impurity (such as：(1-17) segment, 18-31 segments), the sample purity of collection is up to 96%；
It is stationary phase and trifluoroacetic acid solution to select HILIC chromatographic column fillers：Acetonitrile=70:30(V:V) it is mobile phase phase
In conjunction with chromatographic condition, to the peptides of the more Gly sequences being synthetically produced, lack the peptide and epimer (such as D-His) of Gly sequences
There is good separating effect Deng the purity of sample and the impurity of yield is influenced；The sample that purity is 99.6% or more can once be received
Product.It is not high to solve Liraglutide finished product purity in turn, the low problem of yield.
Description of the drawings
Fig. 1 is the mass spectrogram of finished product after purification of the invention；
Fig. 2 is 1/2 figure of the HPLC of Liraglutide crude product of the present invention；
Fig. 3 is 2/2 figure of the HPLC of Liraglutide crude product of the present invention；
Fig. 4 be first time HPLC of the invention after purification Liraglutide HPLC figure；
Fig. 5 be second of HPLC of the present invention after purification Liraglutide HPLC figure.
Specific implementation mode
Specific introduce is made to the present invention below in conjunction with the drawings and specific embodiments.
A method of purifying Liraglutide includes the following steps：
Step 1, Liraglutide crude product pre-process to obtain the thick peptide aqueous solution of Liraglutide；The pre-treatment step includes：
1) Liraglutide crude product is dissolved in acetonitrile solution and obtains the thick peptide aqueous solution of Liraglutide；
2) Liraglutide aqueous solution membrane filtration is taken to remove insoluble granule.It is spare to collect filtrate, as a preferred embodiment,
Filter membrane is 0.22 μm of filter membrane.
Step 2, the first time HPLC purifying of removal segment impurity, segment impurity packet
It includes:H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-
Gln-OH,
H-Ala-Ala-Lys(Pal-γ-Glu)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-
Gly-OH；
First time HPLC purifying specific steps include：Using eight alkyl silane bonded silica gel fillers as stationary phase, with phosphoric acid
For mobile phase A, using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC linear gradient elutions, collects and draws Shandong containing advantageous
The fraction of peptide sample；As a kind of embodiment, mobile phase A uses 0.5% phosphoric acid solution, phosphoric acid solution to add 5ml by 1000ml water
Phosphoric acid is uniformly mixed, and is obtained with triethylamine adjusting pH value to 7.0.
Step 3, with rotary evaporator water bath temperature at 30~35 DEG C, vacuum degree removes part second below -0.09MPa
Nitrile, obtains Liraglutide first step sample solution, and sample solution is alkalinity.
Step 4 removes the sequence for the more Gly being synthetically produced, lacks the sequence of Gly and second of HPLC of epimer
Purifying, including impurity：D-His1、D-Ser8、D-Ala19、D-Glu21、D-Arg30、Endo-Gly31、Endo-Gly29、Des-
Gly16；
Second HPLC purifying the specific steps are：Using HILIC chromatographic column fillers as stationary phase, with trifluoroacetic acid solution with
The mixed liquor of acetonitrile is mobile phase A, and using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC gradient elution, collects and contain
There is the fraction of Liraglutide sample.As a kind of embodiment, mobile phase A is 0.08% trifluoroacetic acid solution：Acetonitrile=70V:30V
Mixed liquor, 0.08% trifluoroacetic acid solution adds trifluoroacetic acid 0.8ml to be mixed by 1000ml water.It should be noted that：Herein
On the basis of make proportioning adjustment arbitrary proportion all within protection scope of the present invention.
Step 5, with rotary evaporator water bath temperature at 30~35 DEG C, vacuum degree removes part second below -0.09MPa
Nitrile obtains Liraglutide second step sample solution, and Liraglutide second step sample solution contains trifluoroacetic acid, and solution is acidity, is used
Ammonium hydroxide tune PH to 6.5.
In order to prove the purification process of the present invention, impurity can be removed, obtain purity height, the Liraglutide of high income at
Product do purification experiment with the Liraglutide crude product sample containing 2.9g, 8g, 32g, 72g with this method, and experimentation is as follows：
Embodiment 1：
Take Liraglutide crude product
Sample treatment：2.9g Liraglutides crude product (crude product weight will be contained：4 grams) sample is dissolved in acetonitrile solution, and it is completely molten
With 0.22 μm of membrane filtration after solution.It is spare to collect the thick peptide aqueous solution of filtered Liraglutide.
First step HPLC purifying
Chromatographic condition：With eight alkyl silane bonded silica gel filler stationary phases (30mm × 250mm, 10 μm) for chromatographic column；With
0.5% phosphoric acid (take 1000ml water, add 5ml phosphoric acid, be uniformly mixed, with triethylamine adjust pH value to 7.0) be mobile phase A；With second
Nitrile is Mobile phase B；Flow velocity is that 20mL is per minute；Detection wavelength is 230nm；Single needle applied sample amount is 0.57g.
Following table gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 90%.With rotary evaporator water bath temperature at 30~35 DEG C,
Vacuum degree removes partial acetonitrile below -0.09MPa.Obtain Liraglutide first step sample solution.
Second step HPLC purifying
Chromatographic condition：With HILIC chromatograph packing materials (30mm × 250mm, 10 μm) for chromatographic column；It is molten with 0.08% trifluoroacetic acid
Liquid (takes 1000ml water to add trifluoroacetic acid 0.8ml):Acetonitrile=70:30(V:V) it is mobile phase A；Using acetonitrile as Mobile phase B；Flow velocity
It is per minute for 20mL；Detection wavelength is 230nm；Applied sample amount is 0.36g.Following table gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 99.6%.With rotary evaporator water bath temperature 30~35
DEG C, vacuum degree removes partial acetonitrile below -0.09MPa, and through reference substance, quantitatively 2.40g containing Liraglutide, yield reach solution
83.0%.
Embodiment 2
Take Liraglutide crude product
Sample treatment：8g Liraglutides crude product (crude product weight will be contained：11.2 grams) it is dissolved in acetonitrile solution, after being completely dissolved
With 0.22 μm of membrane filtration.It is spare to collect filtered thick peptide aqueous solution.
First step HPLC purifying
Chromatographic condition：Using eight alkyl silane bonded silica gel fillers as the chromatographic column of stationary phase (50mm × 250mm, 10 μm)；
With 0.5% phosphoric acid (take 1000ml water, add 5ml phosphoric acid, be uniformly mixed, with triethylamine adjust pH value to 7.0) be mobile phase A；With
Acetonitrile is Mobile phase B；Flow velocity is that 60mL is per minute；Detection wavelength is 230nm；Single needle applied sample amount is 1.6g.Following table elution ladder
Degree is eluted.
Collect the fraction of Liraglutide sample of the purity more than 90%.With rotary evaporator water bath temperature at 30~35 DEG C,
Vacuum degree removes partial acetonitrile below -0.09MPa.Obtain Liraglutide first step sample solution.
Second step HPLC purifying
Chromatographic condition：Using HILIC chromatograph packing materials as the chromatographic column of stationary phase (50mm × 250mm, 10 μm)：With 0.08% 3
Fluoroacetic acid solution (takes 1000ml water to add trifluoroacetic acid 0.8ml):Acetonitrile=70:30(V:V) it is mobile phase A；It is flowing with acetonitrile
Phase B；Flow velocity is that 60mL is per minute；Detection wavelength is 230nm；Applied sample amount is 1.0g.Following table gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 99.6%.With rotary evaporator water bath temperature 30~35
DEG C, vacuum degree removes partial acetonitrile below -0.09MPa；Through reference substance, quantitatively 6.40g containing Liraglutide, yield reach solution
80.0%.
Embodiment 3
Take Liraglutide crude product
Sample treatment：32g Liraglutides crude product (crude product weight will be contained：44.9 grams) it is dissolved in acetonitrile solution, it is completely dissolved
Afterwards with 0.22 μm of membrane filtration.It is spare to collect filtered thick peptide aqueous solution.
First step HPLC purifying
Chromatographic condition：Using eight alkyl silane bonded silica gel fillers as the chromatographic column of stationary phase (100mm × 250mm, 10 μm)；
With 0.5% phosphoric acid (take 1000ml water, add 5ml phosphoric acid, be uniformly mixed, with triethylamine adjust pH value to 7.0) be mobile phase A；With
Acetonitrile is Mobile phase B；Flow velocity is that 200mL is per minute；Detection wavelength is 230nm；Single needle applied sample amount is 6.4g.Following table elution ladder
Degree is eluted.
Collect the fraction of Liraglutide sample of the purity more than 90%.With rotary evaporator water bath temperature at 30~35 DEG C,
Vacuum degree removes partial acetonitrile below -0.09MPa.Obtain Liraglutide first step sample solution.
Second step HPLC purifying
Chromatographic condition：Using HILIC chromatograph packing materials as the chromatographic column of stationary phase (100mm × 250mm, 10 μm)；With 0.08%
Trifluoroacetic acid solution (takes 1000ml water to add trifluoroacetic acid 0.8ml):Acetonitrile=70:30(V:V) it is mobile phase A；It is stream with acetonitrile
Dynamic phase B；Flow velocity is that 200mL is per minute；Detection wavelength is 230nm；Applied sample amount is 4.0g.Following table gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 99.6%.With rotary evaporator water bath temperature 30~35
DEG C, vacuum degree removes partial acetonitrile below -0.09MPa；Through reference substance, quantitatively 26.33g containing Liraglutide, yield reach solution
82.3%.
Embodiment 4
Take Liraglutide crude product
Sample treatment：72g Liraglutides crude product (crude product weight will be contained：101.1 grams) it is dissolved in acetonitrile solution, it is completely dissolved
Afterwards with 0.22 μm of membrane filtration.Collect that filtered thick peptide aqueous solution is spare, chromatogram such as Fig. 2 of Liraglutide crude product, 3 institutes
Show.First step HPLC purifying
Chromatographic condition：Using eight alkyl silane bonded silica gel fillers as the chromatographic column of stationary phase (150mm × 250mm, 10 μm)；
With 0.5% phosphoric acid (take 1000ml water, add 5ml phosphoric acid, be uniformly mixed, with triethylamine adjust pH value to 7.0) be mobile phase A；With
Acetonitrile is Mobile phase B；Flow velocity is that 600mL is per minute；Detection wavelength is 230nm；Single needle applied sample amount is 14.4g；Following table elutes
Gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 90%.With rotary evaporator water bath temperature at 30~35 DEG C,
Vacuum degree removes partial acetonitrile below -0.09MPa.Liraglutide first step sample solution is obtained, first step HPLC is after purification
Chromatogram it is as shown in Figure 4.
Second step HPLC purifying
Chromatographic condition：Using HILIC chromatograph packing materials as the chromatographic column of stationary phase (150mm × 250mm, 10 μm)；With 0.08%
Trifluoroacetic acid solution (takes 1000ml water to add trifluoroacetic acid 0.8ml):Acetonitrile=70:30(V:V) it is mobile phase A；It is stream with acetonitrile
Dynamic phase B；Flow velocity is that 600mL is per minute；Detection wavelength is 230nm；Applied sample amount is 9.0g.Following table gradient is eluted.
Collect the fraction of Liraglutide sample of the purity more than 99.6%.With rotary evaporator water bath temperature 30~35
DEG C, vacuum degree removes partial acetonitrile below -0.09MPa；Through reference substance, quantitatively 58.68g containing Liraglutide, yield reach solution
81.5%.
By 4 embodiments it is found that purifying Liraglutide by this method, the yield higher than 80% can be obtained, and obtain
Liraglutide purity be more than 99.6%, the chromatograms of second step HPLC after purification are as shown in Figure 5.
The present invention provides a kind of method of purifying Liraglutide, can remove the peptide for the more Gly sequences being synthetically produced, lack
Peptide, epimer (such as D-His) impurity of Gly sequences improve and draw Shandong peptide sample purity and yield, and yield is higher than 80%,
The purity of Liraglutide is more than 99.6%.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the invention is not limited in any way above-described embodiment, all to be obtained by the way of equivalent substitution or equivalent transformation
Technical solution is all fallen in protection scope of the present invention.
Claims (8)
1. a kind of method of purifying Liraglutide, which is characterized in that include the following steps：
Step 1, Liraglutide crude product pre-process to obtain the thick peptide aqueous solution of Liraglutide；
Step 2, the first time HPLC purifying of removal segment impurity；
Step 3 removes solvent, obtains Liraglutide first step sample solution；
Step 4, peptide, peptide, second of the HPLC of epimer of scarce Gly sequences for removing the more Gly sequences being synthetically produced are pure
Change；
Step 5 removes solvent and obtains Liraglutide second step sample solution, adjusts pH value to 6.5, it is final to obtain Liraglutide
Sample solution.
2. a kind of method of purifying Liraglutide according to claim 1, which is characterized in that step 1, Liraglutide are thick
Product pre-process to obtain the thick peptide aqueous solution of Liraglutide；The pretreated step includes：
1) Liraglutide crude product is dissolved in acetonitrile solution and obtains the thick peptide aqueous solution of Liraglutide；
2) it takes Liraglutide aqueous solution membrane filtration to remove insoluble granule, it is spare to collect filtrate.
3. a kind of method of purifying Liraglutide according to claim 1, which is characterized in that
Step 2, the first time HPLC purifying of removal segment impurity, the segment impurity include：
H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Le u-Glu-Gly-Gln-OH,
H-Ala-Ala-Lys(Pal-γ-Glu)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH；
First time HPLC purifying specific steps include：It is stream with phosphoric acid using eight alkyl silane bonded silica gel fillers as stationary phase
Dynamic phase A, using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC linear gradient elutions, collects containing Liraglutide sample
The fraction of product.
4. a kind of method of purifying Liraglutide according to claim 3, which is characterized in that
The mobile phase A uses 0.5% phosphoric acid solution, phosphoric acid solution to add 5ml phosphoric acid by 1000ml water, is uniformly mixed, with three second
Amine adjusts pH value and obtains to 7.0.
5. a kind of method of purifying Liraglutide according to claim 1, which is characterized in that step 3 uses rotary evaporation
For device bath temperature at 30~35 DEG C, vacuum degree removes partial acetonitrile below -0.09MPa, obtains Liraglutide first step sample
Solution, sample solution are alkalinity.
6. a kind of method of purifying Liraglutide according to claim 1, which is characterized in that step 4, removal synthesis production
Second of HPLC purifying of the sequence of raw more Gly, the sequence for lacking Gly and epimer；Second of HPLC purifying removal
Impurity includes：D-His1、D-Ser8、D-Ala19、D-Glu21、D-Arg30、Endo-Gly31、Endo-Gly29、Des-Gly16；Institute
State second HPLC purifying the specific steps are：Using HILIC chromatographic column fillers as stationary phase, with trifluoroacetic acid solution and acetonitrile
Mixed liquor is mobile phase A, and using acetonitrile as Mobile phase B, Detection wavelength 230nm carries out HPLC gradient elution, collects and is drawn containing advantageous
The fraction of Shandong peptide sample.
7. a kind of method of purifying Liraglutide according to claim 6, which is characterized in that mobile phase A is 0.08% 3
Fluoroacetic acid solution：Acetonitrile=70V:The mixed liquor of 30V, 0.08% trifluoroacetic acid solution add trifluoroacetic acid 0.8ml by 1000ml water
It is mixed into.
8. a kind of method of purifying Liraglutide according to claim 1, which is characterized in that step 5 uses rotary evaporation
For device bath temperature at 30~35 DEG C, vacuum degree removes partial acetonitrile below -0.09MPa, obtains Liraglutide second step sample
Solution, Liraglutide second step sample solution contain trifluoroacetic acid, and solution is acidity, with ammonium hydroxide tune PH to 6.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810663443.7A CN108794618B (en) | 2018-06-25 | 2018-06-25 | Method for purifying liraglutide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810663443.7A CN108794618B (en) | 2018-06-25 | 2018-06-25 | Method for purifying liraglutide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108794618A true CN108794618A (en) | 2018-11-13 |
| CN108794618B CN108794618B (en) | 2021-08-17 |
Family
ID=64071270
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810663443.7A Active CN108794618B (en) | 2018-06-25 | 2018-06-25 | Method for purifying liraglutide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108794618B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109354622A (en) * | 2018-12-05 | 2019-02-19 | 苏州汇通色谱分离纯化有限公司 | A kind of Suo Malu peptide purification filler special and its purification process |
| CN109456403A (en) * | 2018-12-31 | 2019-03-12 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of synthetic method of Liraglutide |
| CN110540587A (en) * | 2019-08-30 | 2019-12-06 | 杭州诺泰澳赛诺医药技术开发有限公司 | Chromatographic method for effectively improving purification yield of synthetic peptide |
| CN111269309A (en) * | 2018-12-04 | 2020-06-12 | 翰宇药业（武汉）有限公司 | Purification method of GLP-1 analog polypeptide |
| TWI738260B (en) * | 2019-03-25 | 2021-09-01 | 台灣神隆股份有限公司 | Process for purifying liraglutide |
| CN113984911A (en) * | 2020-07-27 | 2022-01-28 | 宁波鲲鹏生物科技有限公司 | Chromatographic method for simultaneously analyzing liraglutide and Boc-liraglutide main chain thereof |
| CN116183752A (en) * | 2022-12-29 | 2023-05-30 | 江苏诺泰澳赛诺生物制药股份有限公司 | Liquid chromatography detection method for polypeptide impurities |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102584982A (en) * | 2012-02-10 | 2012-07-18 | 深圳翰宇药业股份有限公司 | Method for purifying solid-phase synthetic coarse liraglutide |
| CN105017381A (en) * | 2015-07-20 | 2015-11-04 | 吉尔生化（上海）有限公司 | Purification method of liraglutide |
| CN107892717A (en) * | 2017-12-29 | 2018-04-10 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of method of purifying Suo Malu peptides |
| CN107903318A (en) * | 2017-12-29 | 2018-04-13 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of method for purifying Liraglutide |
-
2018
- 2018-06-25 CN CN201810663443.7A patent/CN108794618B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102584982A (en) * | 2012-02-10 | 2012-07-18 | 深圳翰宇药业股份有限公司 | Method for purifying solid-phase synthetic coarse liraglutide |
| CN105017381A (en) * | 2015-07-20 | 2015-11-04 | 吉尔生化（上海）有限公司 | Purification method of liraglutide |
| CN107892717A (en) * | 2017-12-29 | 2018-04-10 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of method of purifying Suo Malu peptides |
| CN107903318A (en) * | 2017-12-29 | 2018-04-13 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of method for purifying Liraglutide |
Non-Patent Citations (1)
| Title |
|---|
| 理查德 J. 辛普森: "《蛋白质组学中的蛋白质纯化手册》", 31 March 2009, 化学化工出版社 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111269309A (en) * | 2018-12-04 | 2020-06-12 | 翰宇药业（武汉）有限公司 | Purification method of GLP-1 analog polypeptide |
| CN111269309B (en) * | 2018-12-04 | 2022-03-08 | 翰宇药业（武汉）有限公司 | Purification method of GLP-1 analog polypeptide |
| CN109354622A (en) * | 2018-12-05 | 2019-02-19 | 苏州汇通色谱分离纯化有限公司 | A kind of Suo Malu peptide purification filler special and its purification process |
| CN109456403A (en) * | 2018-12-31 | 2019-03-12 | 江苏诺泰澳赛诺生物制药股份有限公司 | A kind of synthetic method of Liraglutide |
| TWI738260B (en) * | 2019-03-25 | 2021-09-01 | 台灣神隆股份有限公司 | Process for purifying liraglutide |
| US11459354B2 (en) | 2019-03-25 | 2022-10-04 | Scinopharm Taiwan, Ltd. | Process for purifying liraglutide |
| CN110540587A (en) * | 2019-08-30 | 2019-12-06 | 杭州诺泰澳赛诺医药技术开发有限公司 | Chromatographic method for effectively improving purification yield of synthetic peptide |
| CN113984911A (en) * | 2020-07-27 | 2022-01-28 | 宁波鲲鹏生物科技有限公司 | Chromatographic method for simultaneously analyzing liraglutide and Boc-liraglutide main chain thereof |
| CN116183752A (en) * | 2022-12-29 | 2023-05-30 | 江苏诺泰澳赛诺生物制药股份有限公司 | Liquid chromatography detection method for polypeptide impurities |
| CN116183752B (en) * | 2022-12-29 | 2023-09-19 | 江苏诺泰澳赛诺生物制药股份有限公司 | Liquid chromatography detection method for polypeptide impurities |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108794618B (en) | 2021-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108794618A (en) | A method of purifying Liraglutide | |
| CN108640985B (en) | Method for purifying somaglutide | |
| CN107903318A (en) | A kind of method for purifying Liraglutide | |
| CN107892717A (en) | A kind of method of purifying Suo Malu peptides | |
| CN103613655B (en) | Method for low-cost purification of exenatide | |
| CN101372504B (en) | Method for purifying desmopressin | |
| CN106699847B (en) | Method for purifying hexapeptide at low cost | |
| CN112661815B (en) | Method for purifying Tirzepatide | |
| CN110540587A (en) | Chromatographic method for effectively improving purification yield of synthetic peptide | |
| CN108373499B (en) | A kind of purifying and ionic control method of Teriparatide acetate | |
| CN106167522A (en) | A kind of method of extensive isolated and purified teriparatide (Teriparatide) | |
| CN102219849B (en) | Method for separating and purifying exenatide on large scale | |
| CN109354622A (en) | A kind of Suo Malu peptide purification filler special and its purification process | |
| CN103275209A (en) | Liraglutide preparation method | |
| CN114369142B (en) | Method for purifying desmopressin acetate | |
| CN112724241A (en) | Purification method of abamectin | |
| CN103265630B (en) | The preparation method of Exenatide | |
| CN108610391A (en) | A method of extracting polysaccharide and adenosine from Phellinus fructification | |
| CN111269309B (en) | Purification method of GLP-1 analog polypeptide | |
| CN108822164A (en) | The preparation process of high quality monosialotetrahexose ganglioside sodium | |
| CN109406685B (en) | High performance liquid chromatography method for separating carfilzomib and isomers thereof | |
| CN103694338B (en) | A kind of purification process of glucagon hydrochloride | |
| CN207614488U (en) | Ultrafiltration membrane separating device for saponin(e purification | |
| CN110194758A (en) | A method of the fast separating and purifying Aristolochic Acid compound from caulis aristologhiae manshuriensis | |
| CN111363024A (en) | Method for purifying polypeptide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |