CN107266535A - A kind of purification process of Linaclotide - Google Patents
A kind of purification process of Linaclotide Download PDFInfo
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- CN107266535A CN107266535A CN201710672918.4A CN201710672918A CN107266535A CN 107266535 A CN107266535 A CN 107266535A CN 201710672918 A CN201710672918 A CN 201710672918A CN 107266535 A CN107266535 A CN 107266535A
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- mobile phase
- linaclotide
- hcl buffer
- buffer solutions
- eluent
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- KXGCNMMJRFDFNR-WDRJZQOASA-N linaclotide Chemical compound C([C@H](NC(=O)[C@@H]1CSSC[C@H]2C(=O)N[C@H]3CSSC[C@H](N)C(=O)N[C@H](C(N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N2)=O)CSSC[C@H](NC(=O)[C@H](C)NC(=O)[C@@H]2CCCN2C(=O)[C@H](CC(N)=O)NC3=O)C(=O)N[C@H](C(NCC(=O)N1)=O)[C@H](O)C)C(O)=O)C1=CC=C(O)C=C1 KXGCNMMJRFDFNR-WDRJZQOASA-N 0.000 title claims abstract description 54
- 108010024409 linaclotide Proteins 0.000 title claims abstract description 54
- 229960000812 linaclotide Drugs 0.000 title claims abstract description 53
- 238000000746 purification Methods 0.000 title claims abstract description 11
- 239000003480 eluent Substances 0.000 claims abstract description 25
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- 239000007853 buffer solution Substances 0.000 claims abstract description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 18
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 17
- 238000010828 elution Methods 0.000 claims abstract description 15
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000011068 loading method Methods 0.000 claims abstract description 14
- 239000012043 crude product Substances 0.000 claims abstract description 13
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 13
- 238000005349 anion exchange Methods 0.000 claims abstract description 11
- 238000010612 desalination reaction Methods 0.000 claims abstract description 10
- 239000011780 sodium chloride Substances 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims abstract description 10
- 125000000218 acetic acid group Chemical class C(C)(=O)* 0.000 claims abstract description 7
- 239000007864 aqueous solution Substances 0.000 claims abstract description 6
- 238000004255 ion exchange chromatography Methods 0.000 claims abstract description 5
- 230000005526 G1 to G0 transition Effects 0.000 claims abstract description 4
- 239000012071 phase Substances 0.000 claims description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 229920002981 polyvinylidene fluoride Polymers 0.000 claims description 5
- 238000012856 packing Methods 0.000 claims description 4
- 239000012564 Q sepharose fast flow resin Substances 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 238000004587 chromatography analysis Methods 0.000 claims description 2
- 238000005374 membrane filtration Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 abstract description 10
- 238000000926 separation method Methods 0.000 abstract description 3
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 9
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 206010010774 Constipation Diseases 0.000 description 6
- 230000013872 defecation Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000004506 ultrasonic cleaning Methods 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- 206010000060 Abdominal distension Diseases 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 208000003243 intestinal obstruction Diseases 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 206010000087 Abdominal pain upper Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 108010078321 Guanylate Cyclase Proteins 0.000 description 1
- 102000014469 Guanylate cyclase Human genes 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 244000097724 Mesua ferrea Species 0.000 description 1
- 235000010931 Mesua ferrea Nutrition 0.000 description 1
- 206010029333 Neurosis Diseases 0.000 description 1
- 235000005704 Olneya tesota Nutrition 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 206010034647 Peristalsis visible Diseases 0.000 description 1
- 235000008198 Prosopis juliflora Nutrition 0.000 description 1
- 208000005392 Spasm Diseases 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000011097 chromatography purification Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229940084408 linzess Drugs 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 208000015238 neurotic disease Diseases 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 210000001599 sigmoid colon Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 230000001148 spastic effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 208000022534 visible peristalsis Diseases 0.000 description 1
- 210000004916 vomit Anatomy 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
Abstract
The invention mainly relates to a kind of purification process of Linaclotide, belong to technical field of bioseparation.The method that purification process of the present invention uses ion-exchange chromatography combination high performance liquid chromatography, using anion-exchange column as stationary phase, using Tris HCl buffer solutions as mobile phase A, using the Tris HCl buffer solutions containing NaCl as Mobile phase B, Linaclotide crude product solution is handled using gradient elution method, desalination is carried out to above-mentioned eluent using high performance liquid chromatography afterwards and turns acetic acid salt treatment, using C18 posts as separating medium, after loading, using acetic acid aqueous solution as mobile phase A, gradient elution is carried out by Mobile phase B of acetonitrile, eluent obtains Linaclotide sterling by lyophilized.Purification process of the present invention is simple to operate, and separation costs are low, high income, is adapted to the large-scale production of Linaclotide, the sterling purity obtained by purifying is up to more than 99%, and impurity content is low, with good economical and practical value and is widely applied prospect.
Description
Technical field
The invention belongs to field of pharmaceutical chemistry technology, and in particular to a kind of purification process of Linaclotide.
Background technology
Constipation refers to defecation frequency very little, or a kind of alimentary canal that defecation is not smooth, laborious, difficult, excrement is dry and hard and amount is few
Common sympton.Most patients with chronic constipation show as difficult defecation, and excrement is dry and hard, a couple of days even 1 week ability defecation once, during defecation
Can there are the spastic pain of left abdomen and bearing down, some patients tell that bitter taste, anorexia, abdominal distension, lower abdomen are uncomfortable, exhaust is more or has head
The neurosis shape such as dizzy, headache, tired, but do not weigh typically.With tormina, vomit or the person of having blood in stool, be then considered as urgency
Property constipation caused by intestinal obstructions.General physical examination can often touch the intestinal tube or excrement block of spasm in colon descendens or sigmoid colon position,
But then disappeared after defecation.Intestinal obstruction person then often has abdominal distension, stomachache, visible peristalsis visible intestinal peristalsis and peristaltic wave.
Linaclotide（Linaclotide）It is a kind of guanylate cyclase excitement researched and developed by Ironwood companies of the U.S.
Agent, was approved listing by U.S. FDA, trade name Linzess on the 30th in August in 2012.The medicine is capsule, is mainly used in controlling
Treat constipation IBS (IBS-C) and chronic idiopathic constipation (CIC), be the first treatment with such a mechanism of action just
Secret polypeptide drugs.
The molecular structure of Linaclotide is a small peptide by 14 Amino acid profiles, and sequence is Cys-Cys-Glu-Tyr-
Cys-Cys-Asn-Pro-Ala-Cys-Thr-Gly-Cys-Thr-OH(1-6),(2-10),(5-13)-tris
(disulfide).The drug form, to the unformed powder of canescence, is slightly soluble in water and physiological saline to be white.Linaclotide
Molecular structure be
。
The presence of multipair disulfide bond brings larger difficulty to the purifying of Linaclotide in molecule, is embodied in following side
Face：First when synthesizing Linaclotide, the impurity of a variety of disulfide bond mispairing can be produced, this kind of impurity is difficult to separate with product, is led
Cause final products purity low；Secondly because there is multipair disulfide bond, Linaclotide in stereochemical structure with general polypeptide difference compared with
Greatly, conventional separating medium hardly results in the product of high-purity；More poly is easily finally produced in process of producing product
Body, is one of major impurity of product, and itself is extremely unstable, has a strong impact on color and luster, content and the security of Linaclotide.
Chinese invention patent CN105153284A provides a kind of method that reversed-phase liquid chromatography gradient elution is isolated and purified,
CN103848893A provides a kind of porous graphitic carbon stationary phase and combines the anti-phase liquid containing three-dimensional selective agent cyclodextrin mobile phase
The method of phase chromatographic separation and purification, both approaches are introduced directly into the thick peptide solution of high salt in high performance liquid chromatography system,
It is unfavorable for obtaining pure peptide product steadily in the long term.In addition, carrying out the purifying of Linaclotide using reversed-phased high performace liquid chromatographic
When, multipair disulfide bond present in the molecule can cause its purification effect undesirable, and the impurity such as polymer in product is difficult to remove
Go, influence the purity and content of product.
The content of the invention
For above-mentioned problem of the prior art, the invention provides a kind of purifying process of Linaclotide, handed over by ion
Colour changing spectrometry, uses anion-exchange column for separating medium, a large amount of impurity present in thick peptide is removed, with reference to high performance liquid chromatography
Further desalination and turn acetic acid salt treatment, can solve the above problems well, it is ensured that equipment can for a long time, stably, efficiently
The Linaclotide of high-purity is prepared, is adapted to industrialized production.
To realize above-mentioned technical purpose, the present invention provides a kind of purification process of Linaclotide, comprised the following steps：
1）Linaclotide crude product is added into dissolving in pure water and obtains Linaclotide solution；
2）Linaclotide solution is purified using ion-exchange chromatography, the ion-exchange chromatography is with anion exchange
Post is stationary phase, using Tris-HCl buffer solutions as mobile phase A, is carried out by Mobile phase B of the Tris-HCl buffer solutions containing NaCl
Gradient elution, collects eluent；
3）Desalination is carried out to step 2 gained eluent using high performance liquid chromatography and turns acetic acid salt treatment, the efficient liquid phase
Chromatography is using C18 posts as separating medium, after step 2 gained eluent loading, using acetic acid aqueous solution as mobile phase A, using acetonitrile as
Mobile phase B carries out gradient elution, and eluent obtains Linaclotide sterling by lyophilized.
Pass through 0.22 μm of pvdf membrane filtration treatment before Linaclotide solution loading described in step 2.
Anion exchange column packing described in step 2 is preferably SOURCE 15Q or Q-SepharoseFast Flow,
But it is not limited to both resins.
Anion exchange column packing described in step 2 is most preferably SOURCE 15Q.
Gradient elution described in step 2, elution program is flow velocity 0.5mL/min, in 30 min, the hundred of Mobile phase B
Point than being 0 ~ 50%, after Mobile phase B ratio is adjusted into 100% after sample appearance.
Mobile phase A described in step 2 is the Tris-HCl buffer solutions of pH8.2 ~ 8.8, and its concentration is 80 ~ 120 mmol/
L。
Mobile phase A described in step 2 is preferably pH8.5 100 mmol/L Tris-HCl buffer solutions.
Mobile phase B described in step 2 is pH8.2 ~ 8.8Tris-HCl bufferings containing 0.8 ~ 1.2 mol/L NaCl
Liquid.
Mobile phase B described in step 2 is preferably the pH8.5Tris-HCl buffer solutions containing 1 mol/L NaCl.
The method of the invention can be to using Oxygen in Liquid after synthesis in solid state and positioning oxidation technology route or solid phase coupling
Linaclotide made from the technology path and other method of change is long-term, stably purify.Using method pair of the present invention
Linaclotide is purified, simple to operate, and the separation costs of product are relatively low, high income, is adapted to the large-scale production of Linaclotide,
The sterling purity obtained by purifying is up to more than 99%, and impurity content is low, with good economical and practical value and widely should
Use prospect.
Brief description of the drawings
Fig. 1 are the separation spectrogram of Linaclotide crude product（Linaclotide appearance time about 37min）.
Fig. 2 are Linaclotide ESI-MS spectrograms.
Fig. 3 analyze chromatogram for the HPLC of Linaclotide after purification（Linaclotide appearance time about 20min）.
Embodiment
Embodiment 1
1. the processing of Linaclotide crude product
0.5g Linaclotide crude products are weighed, 10mL pure water is added, well mixed be placed in ultrasonic cleaning instrument carries out ultrasound about
10min, promotes the dissolving of sample, draws the supernatant of mixed system, and insoluble impurities is removed with 0.22 μm of pvdf membrane.
2. the purifying of Linaclotide sample
Using anion-exchange column SOURCE 15Q as separating medium, after crude product solution loading, with 100 containing pH8.5
Mmol/L Tris-HCl buffer solutions are mobile phase A, using the pH8.5Tris-HCl buffer solutions containing 1 mol/L NaCl as flowing
Phase B carries out gradient elution, collects eluent；Loading after mobile phase A rinse-system, ready to balance is first used, flow velocity is 0.5mL/
Mobile phase A and Mobile phase B ratio are adjusted into 50% in min, 30min to be eluted.After Mobile phase B ratio is adjusted after sample appearance
It is rinsed to 100%, mobile phase A ratio is adjusted to 100% and balanced again.
3. the desalination of eluent
Desalination is carried out to eluent using high performance liquid chromatography and turns acetic acid salt treatment, the high performance liquid chromatography is with C18
After post is separating medium, step 2 gained eluent loading, using acetic acid aqueous solution as mobile phase A, carried out by Mobile phase B of acetonitrile
Gradient elution, collects eluent, and Linaclotide sterling is produced after freeze-drying, analyzes the pure of liquid phase measurement Linaclotide sterling
Degree, Linaclotide mean molecule quantity be 1526.74, ESI-MS determine Linaclotide molecular weight 1525.42, after purification profit that
Lip river peptide sterling purity is 99.54%, sees Fig. 1.
Embodiment 2
1. the processing of Linaclotide crude product
0.5g Linaclotide crude products are weighed, 10mL pure water is added, well mixed be placed in ultrasonic cleaning instrument carries out ultrasound about
10min, promotes the dissolving of sample, draws the supernatant of mixed system, and insoluble impurities is removed with 0.22 μm of pvdf membrane.
2. the purifying of Linaclotide sample
Using anion-exchange column Q-SepharoseFast Flow as separating medium, after crude product solution loading, to contain pH8.2
80mmol/L Tris-HCl buffer solutions be mobile phase A, using the pH8.2Tris-HCl buffer solutions containing 0.8mol/L NaCl as
Mobile phase B carries out gradient elution, collects eluent；Loading after mobile phase A rinse-system, ready to balance is first used, flow velocity is
Mobile phase A and Mobile phase B ratio are adjusted into 50% in 0.5mL/min, 30min to be eluted.After after sample appearance by Mobile phase B
Ratio is adjusted to 100% and is rinsed, and mobile phase A ratio is adjusted to 100% and balanced again.
3. the desalination of eluent
Desalination is carried out to eluent using high performance liquid chromatography and turns acetic acid salt treatment, the high performance liquid chromatography is with C18
After post is separating medium, step 2 gained eluent loading, using acetic acid aqueous solution as mobile phase A, carried out by Mobile phase B of acetonitrile
Gradient elution, collects eluent, Linaclotide sterling is produced after freeze-drying, purity is 99.44%.
Embodiment 3
1. the processing of Linaclotide crude product
0.5g Linaclotide crude products are weighed, 10mL pure water is added, well mixed be placed in ultrasonic cleaning instrument carries out ultrasound about
10min, promotes the dissolving of sample, draws the supernatant of mixed system, and insoluble impurities is removed with 0.22 μm of pvdf membrane.
2. the purifying of Linaclotide sample
Using anion-exchange column SOURCE 15Q as separating medium, after crude product solution loading, with 120 containing pH8.8
Mmol/L Tris-HCl buffer solutions are mobile phase A, using the pH8.8Tris-HCl buffer solutions containing 1.2mol/L NaCl as flowing
Phase B carries out gradient elution, collects eluent；Loading after mobile phase A rinse-system, ready to balance is first used, flow velocity is 0.5mL/
Mobile phase A and Mobile phase B ratio are adjusted into 50% in min, 30min to be eluted.After Mobile phase B ratio is adjusted after sample appearance
It is rinsed to 100%, mobile phase A ratio is adjusted to 100% and balanced again.
3. the desalination of eluent
Desalination is carried out to eluent using high performance liquid chromatography and turns acetic acid salt treatment, the high performance liquid chromatography is with C18
After post is separating medium, step 2 gained eluent loading, using acetic acid aqueous solution as mobile phase A, carried out by Mobile phase B of acetonitrile
Gradient elution, collects eluent, Linaclotide sterling is produced after freeze-drying, purity is 99.26%.
Claims (9)
1. a kind of purification process of Linaclotide, it is characterised in that comprise the following steps：
1）Linaclotide crude product is added into dissolving in pure water and obtains Linaclotide solution；
2）Linaclotide is purified using ion-exchange chromatography, the ion-exchange chromatography using anion-exchange column as
Stationary phase, using Tris-HCl buffer solutions as mobile phase A, gradient is carried out by Mobile phase B of the Tris-HCl buffer solutions containing NaCl
Elution, collects eluent；
3）Desalination is carried out to step 2 gained eluent using high performance liquid chromatography and turns acetic acid salt treatment, the efficient liquid phase
Chromatography is using C18 posts as separating medium, after step 2 gained eluent loading, using acetic acid aqueous solution as mobile phase A, using acetonitrile as
Mobile phase B carries out gradient elution, and eluent obtains Linaclotide sterling by lyophilized.
2. according to the method described in claim 1, it is characterised in that pass through before the Linaclotide solution loading described in step 2
0.22 μm of pvdf membrane filtration treatment.
3. according to the method described in claim 1, it is characterised in that the anion exchange column packing described in step 2 is preferably
SOURCE 15Q or Q-SepharoseFast Flow.
4. method according to claim 3, it is characterised in that the anion exchange column packing described in step 2 is most preferably
For SOURCE 15Q.
5. according to the method described in claim 1, it is characterised in that the gradient elution described in step 2, elution program is flow velocity
0.5mL/min, in 30 min, the percentage of Mobile phase B is 0 ~ 50%, after being adjusted to Mobile phase B ratio after sample appearance
100%。
6. according to the method described in claim 1, it is characterised in that the mobile phase A described in step 2 is pH8.2 ~ 8.8
Tris-HCl buffer solutions, its concentration is 80 ~ 120mmol/L.
7. method according to claim 6, it is characterised in that the mobile phase A described in step 2 is preferably the 100 of pH8.5
Mmol/L Tris-HCl buffer solutions.
8. according to the method described in claim 1, it is characterised in that the Mobile phase B described in step 2 is to contain 0.8 ~ 1.2
The Tris-HCl buffer solutions of mol/L NaCl pH8.2 ~ 8.8.
9. method according to claim 8, it is characterised in that the Mobile phase B described in step 2 is preferably to contain 1 mol/
L NaCl pH8.5Tris-HCl buffer solutions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710672918.4A CN107266535A (en) | 2017-08-08 | 2017-08-08 | A kind of purification process of Linaclotide |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710672918.4A CN107266535A (en) | 2017-08-08 | 2017-08-08 | A kind of purification process of Linaclotide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107266535A true CN107266535A (en) | 2017-10-20 |
Family
ID=60077065
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110256534A (en) * | 2018-03-12 | 2019-09-20 | 博瑞生物医药（苏州）股份有限公司 | A kind of purification process of multi-arm anticancer conjugate |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105153284A (en) * | 2015-08-18 | 2015-12-16 | 杭州和泽医药科技有限公司 | Purification method of linaclotide |
| WO2016012938A2 (en) * | 2014-07-22 | 2016-01-28 | Dr. Reddy’S Laboratories Limited | Improved process for preparation of amorphous linaclotide |
| CN106167514A (en) * | 2016-08-29 | 2016-11-30 | 杭州湃肽生化科技有限公司 | The synthesis of a kind of Linaclotide and purification process |
-
2017
- 2017-08-08 CN CN201710672918.4A patent/CN107266535A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016012938A2 (en) * | 2014-07-22 | 2016-01-28 | Dr. Reddy’S Laboratories Limited | Improved process for preparation of amorphous linaclotide |
| CN105153284A (en) * | 2015-08-18 | 2015-12-16 | 杭州和泽医药科技有限公司 | Purification method of linaclotide |
| CN106167514A (en) * | 2016-08-29 | 2016-11-30 | 杭州湃肽生化科技有限公司 | The synthesis of a kind of Linaclotide and purification process |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110256534A (en) * | 2018-03-12 | 2019-09-20 | 博瑞生物医药（苏州）股份有限公司 | A kind of purification process of multi-arm anticancer conjugate |
| CN110256534B (en) * | 2018-03-12 | 2023-08-25 | 高瑞耀业(北京)科技有限公司 | Purification method of multi-arm anticancer conjugate |
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Application publication date: 20171020 |