CN106854230A - A kind of solid phase fragment method synthesizes carbetocin - Google Patents
A kind of solid phase fragment method synthesizes carbetocin Download PDFInfo
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- CN106854230A CN106854230A CN201510897369.1A CN201510897369A CN106854230A CN 106854230 A CN106854230 A CN 106854230A CN 201510897369 A CN201510897369 A CN 201510897369A CN 106854230 A CN106854230 A CN 106854230A
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- Prior art keywords
- resin
- amino
- carbetocin
- overprotection
- solid phase
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- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of solid phase fragment preparation method of carbetocin, it is comprised the following steps：1) proline, cysteine, asparagine (Asn), glutamine, isoleucine, the tyrosine of amino and side chain through overprotection are sequentially connected by carrier of synthesis in solid state resin；2) cysteine sulfydryl protection group is removed；3) in the basic conditions, it is coupled bromo-butyric acid；4) after removing terminal amino group protection group, terminal amino group is coupled cyclization with cysteine side chain carboxyl；5) ring seven peptide fragment resin is cracked；6) glycine and leucine of the amino through overprotection, 7 are sequentially connected by carrier of synthesis in solid state resin) by step 5) gained ring seven peptide fragment is connected on dipeptide resin；8) carbetocin is obtained after cracking.Site between the application selection Pro-Leu, the carbetocin purity that preparation method of the invention is obtained is high.
Description
Technical field
The present invention relates to a kind of preparation method of cyclic peptide, and in particular to the solid phase synthesis process of carbetocin.
Background technology
Carbetocin (Carbetocin) is a kind of long-acting oxytocins nonapeptide class with agonist characteristics of synthesis
Like thing.Single-dose intravenous can be administered immediately after caesarean section under Epidural cavity or lumbar anesthesia, to prevent uterus tension force not
Foot and postpartum haemorrhage.
The clinic and pharmacological property of carbetocin are similar with naturally-produced oxytocins.As oxytocins, card
Shellfish oxytocin is combined with the ocytocin receptor of uterine smooth muscle, causes the Rythmic contractions characteristic in uterus, in original receipts
On the basis of contracting, increase its frequency and increase uterus tension force.Under non pregnant state, the ocytocin receptor in uterus contains
Amount is very low, increases in gestation, and peak is reached during childbirth.Therefore carbetocin does not have to the uterus of non-pregnant
Effect, but there is effective uterine contractile to act in the uterus in the uterus and firm production to gestation.
Research shows that Single-dose intravenous give carbetocin 100 immediately after caesarean section under Epidural cavity or lumbar anesthesia
Ug, in terms of the not enough postpartum haemorrhage with reduction of prevention uterus tension force, carbetocin is substantially better than placebo.
The early stage in postpartum gives carbetocin can also promote the restoration of old ways in uterus.
Carbetocin has following structure：
It is expressed as with amino acid abbreviations：
The preparation method early stage mainly liquid phase synthesis process of carbetocin and the like, complex operation, no
It is not high beneficial to industrial production, application value；In terms of the patent synthesis that carbetocin is declared abroad at present
Seldom, it is the synthetic method of solid liquid phase combination including an ES2115543 for company of Spain, main side
Method is：Conventional solid Peptide systhesis are used, is obtained using HOBt/DIC systems
4-chlorobutyl-Tyr (OMe)-Ile-Gln-Asn-Cys (Trt)-Pro-Leu-Gly-PAL-Nle-pMBHA, Ran Houyong
TFA/1-Dodecanethiol/H2O=8:1:1 (the note of cracking 2 hours：1-Dodecanethiol lauryl mercaptans),
Linear peptide 4-chlorobutyl-Tyr (OMe)-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2 is obtained, linear peptides are used
1：1 acetonitrile and water as solvent, pH=9 is adjusted with 1M NaOH, and cyclisation obtains carbetocin.Used by it
Alkali also includes LiOH, NaHCO3, DIEA, DMAP.
Czech patents CS：8605461, first cracked again with solid-phase peptide synthesis synthesis peptide resin and obtained
Z-Ile-Gln-Asn-Cys(Bzl)-Pro-Leu-Gly-NH2, then hydrogenate and obtain
Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2Reacted with 4-Bromobutyric acid again, obtained
Ile-Gln-Asn-Cys (C3H6COOH)-Pro-Leu-Gly-NH2 reacts with X-Tyr (OMe)-OH again, takes off
Protection, cyclisation obtains carbetocin.
In the method for these patents description, cyclization, the liquid phase cyclization of domestic patent are all carried out with liquid phase process
Also Hangzhou and brocade patent (application number：201010560715.4), the patent (Shen of the bright pharmacy of Shanghai Su Hao ease
Please number：201110001400.0) and Suzhou heavenly steed patent (application number 201510001735.0), this side
Method needs reaction raw materials to be reacted in extremely dilute solution, and side reaction is more, and a large amount of solvents are needed in large-scale production,
A large amount of waste liquids are produced immediately.The present invention utilizes solid phase pseudo-dilution principle, develops the efficient cyclisation method of solid phase, instead
Foreshortened to 2~3 hours between seasonable, waste reaction solution is reduced to liquid reactive less than 1/10.And can effectively reduce pair
Reaction, improves thick peptide purity, and then improve yield.
There are many to be also cyclized using solid phase method in domestic patent, can into amido link or first into thioether bond according to elder generation
It is divided into two classes：The first kind be first by bromobutyric acid by acid amides be coupled with the tyrosine amino coupled of peptide chain, then use
Alkali (such as DIPEA, NMM, LiCl, DMAP) effect is lower to be eliminated hydrogen bromide and forms thioether bond and cyclization.Adopt
Have with the patent of the method Shenzhen writing brush space medicine company the surge peptide of patent (application number 200910106889.0) and Hangzhou it is special
Sharp (application number 201410461695.3)；The another kind of sulfydryl for being first to be connected to the butyric acid of carboxy protective cysteine
On, butyric acid carboxyl-protecting group is removed after being coupled all residues, then be coupled and the tyrosine on peptide chain by acid amides
Amino coupled cyclization, the patent (application number for having the strong unit's medicine company in Shenzhen using the patent of the method
201010544419.5), the patent (application number 201110151928.6) of Chengdu sage promise science and technology, the patent of the triumphant profit in Wuxi
(application number 201210255959.0), the patent (application number 201410331088.5) of Chengdu Mt. Tiantai medicine company.
The Patent Domestic being coupled using fragment also has two.Safe patent (the application number of Hangzhou promise
201310412014.X), individually using the glutamine of carbon teminal an as fragment, remaining 8 residue is used as one
Fragment, cyclisation is carried out using liquid phase process.Hainan it is double into patent (application number 201410076731.4) use
Using the Pro-Leu-Gly of carbon teminal an as fragment, remaining 6 residue as a fragment, use bromo-butyric acid for
Butyric acid residue raw material, the alkali of solid phase cyclization uses DIPEA, TEA or LiOH.
The method for carrying out cyclization with liquid phase process needs reaction raw materials to be reacted in extremely dilute solution, and side reaction is more,
A large amount of solvents are needed in large-scale production, a large amount of waste liquids are produced immediately.The present invention utilizes solid phase pseudo-dilution principle,
The efficient cyclisation method of solid phase is developed, the reaction time foreshortens to 2~3 hours, and waste reaction solution is reduced to liquid reactive
Less than 1/10.And side reaction can be effectively reduced, thick peptide purity is improved, and then improve yield.
Synthesized using fragment, can solve to be coupled the crude product purity for causing one by one in order not high.Fragment method can
Have at three with the site for selecting segmentation, be three sites between Cys-Pro-Leu-Gly.Selection Cys-Pro it
Between (such as 201410076731.4), then Cys for fragment starting connection amino acid, due to Fmoc-Cys (Mmt)-OH
Easily there is racemization when being coupled with Wang resins, Cys enantiomeric impurities is significantly increased (see comparative example 1),
And Cys enantiomeric impurities are very big on purifying influence, fine peptide yield can be greatly reduced.Between selection Leu-Gly
Site (such as 201310412014.X), then an independent glycine is used as fragment, when coupling can be caused to dock, reaction
Steric hindrance is larger, glycine impurity is lacked in product and is raised, and is made troubles to purifying.Therefore the application selects Pro-Leu
Between site.
The content of the invention
One aspect of the invention is related to a kind of solid phase fragment preparation method of carbetocin, and it is comprised the following steps：
1) with synthesis in solid state resin as carrier, by connecting shielded amino acid, amino protecting group is removed, then
The method for connecting shielded amino acid is sequentially connected proline (Pro), the half Guang ammonia of amino and side chain through overprotection
Sour (Cys), asparagine (Asn), glutamine (Gln), isoleucine (Ile), tyrosine (Tyr),
Obtain linear hexapeptide fragment resin of the terminal amino group through overprotection；
2) cysteine sulfydryl protection group is removed, the hexapeptide fragment resin of exposed sulfydryl is obtained；
3) in the basic conditions, be coupled bromo-butyric acid, obtain heptapeptide fragment resin (heptapeptide be above-mentioned hexapeptide and
The conjugate of bromo-butyric acid)；
4) after removing terminal amino group protection group, under condensing agent effect, terminal amino group and cysteine side chain carboxyl
Coupling cyclization, obtains ring seven peptide fragment resin；
5) ring seven peptide fragment resin is cracked, ring seven peptide fragment is obtained；
6) with synthesis in solid state resin as carrier, by connecting shielded amino acid, amino protecting group is removed, then
The method for connecting shielded amino acid is sequentially connected glycine (Gly) and leucine of the amino through overprotection
(Leu) dipeptide resin, is obtained；
7) by step 5) gained ring seven peptide fragment be connected on dipeptide resin, obtain carbetocin peptide resin；
8) carbetocin peptide resin is cracked, carbetocin is obtained.
Further, wherein the amino and side chain are selected from Fmoc-Pro-OH through the proline of overprotection；It is described
Amino and side chain are selected from Fmoc-Cys (Mmt)-OH through the cysteine of overprotection；The amino and side chain are by protecting
The asparagine of shield is selected from Fmoc-Asn (Trt)-OH；The amino and side chain are selected from through the glutamine of overprotection
Fmoc-Gln(Trt)-OH；The amino and side chain are selected from Fmoc-Ile-OH through the isoleucine of overprotection；It is described
Amino and side chain are selected from Fmoc-Tyr (Me)-OH through the tyrosine of overprotection；The amino through overprotection sweet ammonia
Acid is selected from Fmoc-Gly-OH；The amino is selected from Fmoc-Leu-OH through the leucine of overprotection.
Further, step 1) and/or step 7) described in solid phase synthesis process be Fmoc solid-phase polypeptides close
Into method, the coupling agent of selection is DIPCDI+A or DIPEA+A+B, and wherein A is selected from HOBt or HOAt,
B is selected from one or more in PyBOP, PyAOP, HATU, HBTU, TBTU；Preferably, it is coupled
The ratio of each composition is calculated as DIPCDI with molar ratio in agent:A=1.1~1.5:1.0~1.4,
DIPEA:A:B=1.8~2.2:1.0~1.4:0.95~1.05；More preferably DIPCDI:A=1.3:1.2,
DIPEA:A:B=2.0:1.2:1.0.
Further, step 2) removing reagent be trifluoroacetic acid and tri isopropyl silane organic (dichloromethane)
Solution, trifluoroacetic acid concentration is 2~5%, and tri isopropyl silane concentration is 3~8%.
Further, step 3) used by alkali be DIPEA, TEA, NMM, NEM；Step 3) it is described
Solvent is preferably DMF or 1-METHYLPYRROLIDONE, and reaction end uses Ellman reaction detections.
Further, step 4) in cyclization used by condensing agent for TCTU and DIPEA combination, preferably
Ground, the ratio of the two is 1:0.8~1.2.
Further, step 1) used by resin be wang resins, 2-CTC resins, step 6) used by
Resin is Rink Amide resins, Rink Amide-AM resins, Rink Amide-MBHA resins, it is preferable that
Step 1) be wang resins, step 6) be Rink Amide resins, step 6) and described in resin substitution degree
It is 0.2~0.9mmol/g, more preferably 0.4~0.6mmol/g.
Further, step 5) and/or step 8) obtain products therefrom after, also including purification step, preferred institute
State purification step and be selected from recrystallization, reversed phase high-pressure liquid phase process；It is highly preferred that recrystallization method is that thick peptide is molten
In THF, MTBE is then added thereto to.
Further, step 5) lytic reagent that uses is TFA：TIS：H2O=85~95:2~8:2~8 (V:V).
Specific embodiment
Embodiment 1 prepares Fmoc-Pro-Wang resins
Weigh Wang resins 11.949g (13.9mmol) that substitution degree is 1.166mmol/L, DMF washings 2
It is secondary, with DMF swellable resins 30 minutes.Weigh DMAP 0.183g (1.39mmol) add reaction column in.Claim
29.69g (83.4mmol) Fmoc-Pro-OH, plus appropriate DCM dissolvings are taken, ice-water bath cooling is lower to be added
8.7ml (55.6mmol) DIPCDI, by this solution addition reaction column after activating 3 minutes, nitrogen bubble reacts 3h.
Reaction solution is extracted, DMF is washed 3 times, plus 23ml pyridines and 28ml acetic anhydrides carry out capping overnight.Extract
Reaction solution, DMF is washed 3 times, and methyl alcohol shrinkage resin 3 times, dry adsorbent obtains Fmoc-Pro-Wang trees
Fat 22.51g, detection substitution degree is 0.627mmol/g.
The preparation of the heptapeptide cyclic peptide of embodiment 2
Fmoc-Pro-Wang resins 15.959g (10mmol) of the gained of embodiment 1 are weighed, solid phase reaction is added to
In post, washed with DMF 2 times, after DMF swellable resins 30 minutes, DBLK deprotection 6min+8min,
DMF is washed 6 times.Weigh 18.47g (30mmol) Fmoc-Cys (Mmt)-OH and 4.46g (33mmol) HOBT
With DMF/DCM (V:V=1:1) dissolve, after 5.2mL (36mmol) DIPCDI activation 3min is added under ice-water bath,
Mixed liquor is added in reaction column, room temperature reaction 2 hours, reaction end is detected with ninhydrin.
Reaction terminates, and with DMF washing resins 3 times, adds DBLK deprotections 5min+7min, DMF to wash
Wash resin 6 times, ninhydrin detection resin has color.Weigh 17.90g (30mmol) Fmoc-Asn (Trt)-OH and
4.46g (33mmol) HOBT DMF/DCM (V:V=1:1) dissolve, 5.2mL (36mmol) is added under ice-water bath
After DIPCDI activation 3min, mixed liquor is added in reaction column, room temperature reaction 2 hours, is examined with ninhydrin
Survey reaction end.
Coupling Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH, Fmoc-Tyr (Me)-OH are obtained successively in the same way
To full guard hexapeptide resin
Removing Mmt protection groups：With DCM washing resins 6 times, by DCM：TFA：TIS=93:2:5 remove-insurance
Shield liquid is added in solid phase reaction post, after nitrogen air-blowing is reacted 10 minutes, is taken out, and adds deprotection liquid nitrogen tympanites
Solid/liquid/gas reactions 10 minutes, take out, and are repeated two more times deprotection reaction.
Coupling 4- bromo-butyric acids：16.7g (100mmol) 4- bromo-butyric acids are weighed, reaction is added after being dissolved with appropriate DCM
In post, then add 12.9g (100mmol) DIPEA, nitrogen bubble reacts 0.5 hour.With DTNB detections reaction eventually
Point.
Solid phase cyclization：With DMF washing resins 3 times, DBLK deprotections 5min+7min, DMF is added to wash
Wash resin 3 times, DCM：TFA：TIS=93:2:5 solution are washed 3 times, and DMF washs three times, ninhydrin again
Detection resin has color.12.9 are added after weighing 35.5g (100mmol) TCTU, plus DMF dissolvings
G (100mmol) DIPEA, is added in reaction column after stirring and dissolving, and nitrogen bubble reaction 2h is detected with ninhydrin
Reaction end.
Reaction terminates, and drains reaction solution, and DMF washing resins 3 times drain liquid.Methyl alcohol shrinkage resin 3 times,
Peptide resin vacuum drying obtains heptapeptide cyclic peptide resin 25.12g.
Heptapeptide cyclic peptide resin is added in 500ml there-necked flasks, nitrogen protection.Add the TFA for preparing in advance：
TIS：H2O=90:5:5(V:V) 250ml, room temperature reaction 2 hours filters resin, collects filtrate.With a small amount of
TFA washing resins, merging filtrate.Filtrate is slowly added to be precipitated in 2500ml ice ether, is centrifuged, ether
Washing 2 times, drying under reduced pressure obtains 8.0 grams of thick peptide, HPLC purity 88.3%.
By gained 8.0g heptapeptide cyclic peptide thick peptide 40ml THF stirring and dissolvings, 240ml MTBE are added dropwise while stirring,
After ice-water bath is stirred 1 hour, filtering, filter cake vacuum drying obtains heptapeptide cyclic peptide for white solid 7.3g, HPLC
Purity 98.1%.Synthesis yield 89.3%.
Embodiment 3：The preparation of carbetocin peptide resin
Rink Amide resins 5.40g (3.0mmol) that substitution degree is 0.557mmol/g is weighed, solid phase is added to
In reaction column, washed with DMF 2 times, after DMF swellable resins 30 minutes, DBLK deprotections
6min+8min, DMF are washed 6 times.Weigh 2.68g (9mmol) Fmoc-Gly-OH and
1.34g (9.9mmol) HOBT DMF dissolve, and add 1.55mL (10.8mmol) DIPCDI activation 3min
Afterwards, mixed liquor is added in reaction column, room temperature reaction 2 hours, reaction end is detected with ninhydrin.
Reaction terminates, and with DMF washing resins 3 times, adds DBLK deprotections 5min+7min, DMF to wash
Wash resin 6 times, ninhydrin detection resin has color.Weigh 3.18g (9mmol) Fmoc-Leu-OH and
1.34g (9.9mmol) HOBT DMF/DCM (V:V=1:1) dissolve, 1.55 are added under ice-water bath
After mL (10.8mmol) DIPCDI activation 3min, mixed liquor is added in reaction column, room temperature reaction 2 hours,
Reaction end is detected with ninhydrin.
Reaction terminates, and with DMF washing resins 3 times, adds DBLK deprotections 5min+7min, DMF to wash
Wash resin 6 times, ninhydrin detection resin has color.Weigh 7.3g (8.9mmol) heptapeptide cyclic peptide and
1.34g (9.9mmol) HOAT DMF/DCM (V:V=1:1) dissolve, 2.86g (8.9mmol) is added under ice-water bath
After TBTU and 3.1ml (18mmol) DIPEA activation 3min, mixed liquor is added in reaction column, room temperature is anti-
Answer 2 hours, reaction end is detected with ninhydrin.
Reaction terminates, and drains reaction solution, and DMF washing resins 3 times drain liquid.Methyl alcohol shrinkage resin 3 times,
Peptide resin is vacuum dried and obtains g carbetocins peptide resin 7.72, resin weightening 2.32g, theoretical weight gain 2.30g,
Rate of body weight gain 101%.
Embodiment 4：The preparation of carbetocin fine peptide
7.72 grams of the carbetocin peptide resin that embodiment 3 is obtained is added in 125ml there-necked flasks, and nitrogen is protected
Shield.Add the TFA for preparing in advance：TIS：H2O=90:5:5(V:V) 78ml, room temperature reaction 2 hours,
Filtering resin, collects filtrate.With a small amount of TFA washing resins, merging filtrate.Filtrate is slowly added to 780ml ice
Ether in precipitation, centrifugation, ether wash 2 times, drying under reduced pressure obtains 2.83 grams of thick peptide, HPLC purity 96.23%.
Prepared by high pressure liquid phase and purified, lyophilized to obtain carbetocin fine peptide 2.53g, purity 99.12% is maximum single miscellaneous
0.15%.Theoretical yield 2.96g, total recovery 85.5%.
Comparative example：Prepare 6 cyclic peptide
Fed intake 12.49g Wang resins, and Fmoc-Cys (Mmt)-Wang resins are prepared according to the method for embodiment 1,
Resin 29.4g is obtained, substitution degree is 0.26mmol/g.Method according to embodiment 2 is coupled successively
Fmoc-Asn (Trt)-OH, Fmoc-Gln (Trt)-OH, Fmoc-Ile-OH, Fmoc-Tyr (Me)-OH, removing
Mmt protection groups, are coupled 4- bromo-butyric acids, and solid phase cyclization is detected, thick peptide purity 64% after cracking without recrystallization,
Cys racemizations impurity 14%.There is notable difference with the thick peptide purity 88.3% of fragment in embodiment 2.
Claims (9)
1. the solid phase fragment preparation method of a kind of carbetocin, it is comprised the following steps：
1) with synthesis in solid state resin as carrier, by connecting shielded amino acid, amino protecting group is removed, then
The method for connecting shielded amino acid is sequentially connected proline (Pro), the half Guang ammonia of amino and side chain through overprotection
Sour (Cys), asparagine (Asn), glutamine (Gln), isoleucine (Ile), tyrosine (Tyr),
Obtain linear hexapeptide fragment resin of the terminal amino group through overprotection；
2) cysteine sulfydryl protection group is removed, the hexapeptide fragment resin of exposed sulfydryl is obtained；
3) bromo-butyric acid in the basic conditions, is coupled, heptapeptide fragment resin is obtained；
4) after removing terminal amino group protection group, under condensing agent effect, terminal amino group and cysteine side chain carboxyl
Coupling cyclization, obtains ring seven peptide fragment resin；
5) ring seven peptide fragment resin is cracked, ring seven peptide fragment is obtained；
6) with synthesis in solid state resin as carrier, by connecting shielded amino acid, amino protecting group is removed, then
The method for connecting shielded amino acid is sequentially connected glycine (Gly) and leucine of the amino through overprotection
(Leu) dipeptide resin, is obtained；
7) by step 5) gained ring seven peptide fragment be connected on dipeptide resin, obtain carbetocin peptide resin；
8) carbetocin peptide resin is cracked, carbetocin is obtained.
2. the solid phase fragment preparation method of carbetocin according to claim 1, wherein the amino and
Side chain is selected from Fmoc-Pro-OH through the proline of overprotection；The amino and side chain through overprotection cysteine
Selected from Fmoc-Cys (Mmt)-OH；The amino and side chain are selected from through the asparagine of overprotection
Fmoc-Asn(Trt)-OH；The amino and side chain are selected from Fmoc-Gln (Trt)-OH through the glutamine of overprotection；
The amino and side chain are selected from Fmoc-Ile-OH through the isoleucine of overprotection；The amino and side chain are through overprotection
Tyrosine be selected from Fmoc-Tyr (Me)-OH；The amino is selected from Fmoc-Gly-OH through the glycine of overprotection；
The amino is selected from Fmoc-Leu-OH through the leucine of overprotection.
3. the solid phase fragment preparation method of the carbetocin according to any one of claim 1 or 2, step
1) and/or step 7) described in solid phase synthesis process be Fmoc solid-phase peptide synthesis, the coupling of selection
Agent is DIPCDI+A or DIPEA+A+B, wherein A be selected from HOBt or HOAt, B be selected from PyBOP,
One or more in PyAOP, HATU, HBTU, TBTU；Preferably, in coupling agent each composition ratio
Example is calculated as DIPCDI with molar ratio:A=1.1~1.5:1.0~1.4,
DIPEA:A:B=1.8~2.2:1.0~1.4:0.95~1.05；More preferably DIPCDI:A=1.3:1.2,
DIPEA:A:B=2.0:1.2:1.0.
4. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-3, step 2)
Removing reagent be trifluoroacetic acid and tri isopropyl silane organic (dichloromethane) solution, trifluoroacetic acid concentration
It is 2~5%, tri isopropyl silane concentration is 3~8%.
5. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-4, step 3)
Alkali used is DIPEA, TEA, NMM, NEM；Step 3) solvent is preferably DMF or N- methyl
Pyrrolidones, reaction end uses Ellman reaction detections.
6. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-5, step 4)
In cyclization used by condensing agent be the combination of TCTU and DIPEA, it is preferable that the ratio of the two is
1:0.8~1.2.
7. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-6, step 1)
Resin used be wang resins, 2-CTC resins, step 6) used by resin for Rink Amide resins,
Rink Amide-AM resins, Rink Amide-MBHA resins, it is preferable that step 1) it is wang resins,
Step 6) be Rink Amide resins, step 6) described in resin substitution degree be 0.2~0.9mmol/g, it is more excellent
Select 0.4~0.6mmol/g.
8. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-7, step 5)
And/or step 8) obtain product after, also including purification step, preferably described purification step be selected from recrystallization, it is anti-phase
High pressure liquid phase process；It is highly preferred that recrystallization method is that thick peptide is dissolved in into THF, MTBE is then added thereto to.
9. the solid phase fragment preparation method of the carbetocin according to claim any one of 1-8, step 5)
The lytic reagent for using is TFA：TIS：H2O=85~95:2~8:2~8 (V:V).
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| CN110997697A (en) * | 2017-08-11 | 2020-04-10 | 辉凌公司 | Process for preparing pharmaceutical composition |
| CN108047323A (en) * | 2017-12-20 | 2018-05-18 | 深圳羽众医药科技有限公司 | A kind of solid phase segment method synthesis GpTx-1 and the like and synthetic method |
| CN113801199A (en) * | 2020-06-15 | 2021-12-17 | 深圳翰宇药业股份有限公司 | All-solid-phase synthesis method of carbetocin |
| CN113801199B (en) * | 2020-06-15 | 2024-03-05 | 深圳翰宇药业股份有限公司 | All-solid-phase synthesis method of carbetocin |
| CN112028973A (en) * | 2020-08-06 | 2020-12-04 | 山东辰龙药业有限公司 | Resin-carbetocin cracking device |
| CN112028973B (en) * | 2020-08-06 | 2024-05-07 | 山东辰龙药业有限公司 | Resin-carbetocin pyrolysis device |
| CN112592382A (en) * | 2020-11-27 | 2021-04-02 | 安徽大学 | Resin, preparation method thereof and application thereof in preparation of head-tail cyclic peptide |
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