CN101215324B - Exenatide short peptide simulation peptide and application thereof in preparing medicament for curing diabetes - Google Patents
Exenatide short peptide simulation peptide and application thereof in preparing medicament for curing diabetes Download PDFInfo
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- CN101215324B CN101215324B CN2007103003181A CN200710300318A CN101215324B CN 101215324 B CN101215324 B CN 101215324B CN 2007103003181 A CN2007103003181 A CN 2007103003181A CN 200710300318 A CN200710300318 A CN 200710300318A CN 101215324 B CN101215324 B CN 101215324B
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Abstract
The invention provides an exenatide short peptide and imitation peptide, which transforms the structure of exenatide, and is a novel compound. The invention further discloses a preparation process of exenatide short peptide and imitation peptide, which is suitable to be produced in industrialization. The invention further discloses an application on the preparation therapeutic medicaments of diabetes.
Description
Technical field
The present invention discloses a kind of Exenatide small peptide simulating peptide and preparation method thereof, also discloses its application in the preparation Remedies for diabetes simultaneously, belongs to biological pharmacy technical field.
Background technology
The Exenatide that the present invention relates to (hereinafter to be referred as: Exendin-4) be excretory one peptide species in the Gila monster sialisterium of Southwestern United Stares, contain 39 amino acid, glucagon-like-peptide-1 (glucagon-likepeptidel with the people, GLP-1) 53% homology is arranged, the height affinity is arranged and have similar physiological function to the GLP-1 acceptor.
Exendin-4 has pharmacological actions such as the insulin secretion of promotion, glucagon suppression release, triglyceride reducing level and delay gastric acid secretion activity, and Exendin-4 can also promote the beta Cell of islet hyperplasia, stimulates β cell new life and suppress the apoptotic function of β in addition.In Mammals, can resist the hydrolysis of dipeptidyl peptidase.Transformation period in its blood plasma is longer than GLP-1.Therefore have a lot of potential advantages than GLP-1 aspect the treatment diabetes B.
The Exendin-4 of synthetic is developed as the novel blood sugar lowing medicine by Amylin company and the joint research of EliLilly company at present, by the drugs approved by FDA listing, be used for the fully diabetes B patient of controlling blood sugar of N1,N1-Dimethylbiguanide, sulfonylurea or N1,N1-Dimethylbiguanide and sulfonylurea combined utilization in April, 2005.
Novel ofhypoglycemic medicine for the shorter sequence that obtains more to have pharmaceutical use, we transform the structure of Exendin-4, designed short simulating peptide sequence, and introduced the amino acid that β or γ replace, and their activity and stability measured, obtained many in the sequences that keep having on original active basis better stability.
Summary of the invention
The present invention discloses a kind of Exendin-4 small peptide simulating peptide, is that the structure of Exendin-4 is transformed, and is a kind of new compound.
The invention also discloses the preparation method of Exendin-4 small peptide simulating peptide, be applicable to suitability for industrialized production.
The invention also discloses provides the application of Exendin-4 small peptide simulating peptide in the preparation Remedies for diabetes.
The structure of Exendin-4 small peptide simulating peptide is as follows:
| Exendin-4 small peptide simulating peptide | Aminoacid sequence |
| E1 | Gln-Pro-Ser-Val-Gly-Met-Lys-Pro-Ser-Pro-Arg-His |
| E2 | Ser-Val-Ser-Val-Gly-Met-Lys-Pro-Ser-Pro-Arg-Pro |
| E3 | Ser-Val-Phe-Val-Gly-Met-Lys-Pro-Ser-Pro-Ser-Pro |
| E4 | Ser-Val-Ser-Gly-Gly-Met-Lys-Pro-Ser-Pro-Arg-Lys |
| Exendin-4 small peptide simulating peptide | Aminoacid sequence |
| E5 | Ser-Val-Ser-Val-His-Met-Lys-Pro-Ser-Pro-Arg-Pro |
| E6 | Gln-Pro-Ser-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-His |
| E7 | Ser-Val-Ser-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-Pro |
| E8 | Ser-Val-Phe-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Ser-Pro |
| E9 | Ser-Val-Ser-Gly-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-Lys |
| E10 | Ser-Val-Ser-Val-His-Met-β-Lys-Pro-Ser-Pro-Arg-Pro |
| E11 | Gln-Pro-Ser-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-His |
| E12 | Ser-Val-Ser-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-Pro |
| E13 | Ser-Val-Phe-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Ser-Pro |
| E14 | Ser-Val-Ser-Gly-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-Lys |
| E15 | Ser-Val-Ser-Val-His-Met-γ-Lys-Pro-Ser-Pro-Arg-Pro |
The present invention mainly transforms the structure of Exendin-4, has designed the small peptide simulating peptide:
The core texture of small peptide simulating peptide is X-Met-Lys-Pro-Ser-Pro-Y, and wherein X can be Gln-Pro-Ser-Val-Gly, or Gln wherein replaces with Ser; Or Pro replaces with Val; Or Ser replaces with Phe, or Val replaces with Gly, or Gly replaces with His.Y can be Arg-His, or Arg replaces with Ser, or His replaces with Pro or Leu.
The structure of Exendin-4 small peptide simulating peptide of the present invention also comprises wherein Lys with its corresponding beta-amino acids, the displaced aminoacid sequence of gamma-amino acid.
The preparation method of Exendin-4 small peptide simulating peptide adopts the common Fmoc solid phase method of peptide synthesis synthetic among the present invention:
Raw material and reagent: connecing peptide resin is Rink Amide mbha resin.
Amino acid derivative is: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Lys (Boc)-OH, Fmoc-Phe-OH, Fmoc-Arg (pbf)-OH, Fmoc-Val-OH, Fmoc-Met-OH, Fmoc-Gln (Trt)-OH, Fmoc-His (Trt)-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-γ-Lys (Boc)-OH.
The linked reaction condensing agent is: benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.
Cutting reagent is: trifluoroacetic acid.
Processing step is as follows: resin is used DMF swelling 30 minutes, and deprotection is 20 minutes then, washs with DMF; add amino acid derivative and condensing agent in the reactor, room temperature reaction 2 hours, washing; deprotection adds amino acid derivative and condensing agent again, repeats to connecting all amino acid.Add cutting reagent in the reactor again, room temperature reaction 1 hour, after-filtration, filtrate is precipitated in ether, gets crude product.
The purifying of thick product: adopt high performance liquid chromatography to carry out purifying.Elutriant is a trifluoroacetic acid aqueous solution, acetonitrile.Flow velocity is 20ml/min, and the monitoring wavelength is 214nm.
Exendin-4 small peptide simulating peptide among the present invention is also measured respectively its activity, vitro stability, and their pharmacological effect has been carried out preliminary study.The result shows that these small peptide simulating peptide are active suitable with Exendin-4 with external anti-dipeptidyl peptidase enzyme stability, and this invention provides certain foundation for the new peptide medicament of the more simple anti-type ii diabetes of development structure.
Pharmacodynamic experiment according to the Exendin-4 active isomer shows to have the effect of remarkable reduction laboratory animal blood sugar:
Experimental example 1
Cell proliferation experiment
1, the preparation of cell suspension
Rat is put to death, open the abdominal cavity and take out pancreas, add Hanks liquid, the pancreas tissue is shredded adding collagenase vibration 5 minutes, clean the back with Hanks and add Krebs-Ringer bicarbonate damping fluid, the collection pancreas islet. the pancreas islet of collecting is washed 1 time with PBS, be incubated 20 minutes with the PBS salts solution in 37 degrees centigrade. then pancreas islet is blown and beaten into individual cells, cell is positioned in the ice bath, adds nutrient solution, treat that cell attachment can be used for experiment.
2, reaction system
In 96 orifice plates, every porocyte concentration is 10 with the β cell inoculation
7About, the medicine that adds gradient concentration was cultivated after 68 hours, added MTT again, continue to cultivate 4 hours. take out 96 orifice plates then, in 2500r centrifugal 5 minutes, abandon supernatant, every hole adds 100 microlitre dimethyl sulfoxide (DMSO), measures absorption value down in the 570nm wavelength, calculates proliferation rate.
Cell proliferation experiment result is as shown in table 1:
Table 1 cell proliferation experiment
| Group | SR (%) | |||
| 40μM | 80μM | 100μM | 200μM | |
| Exendin-4 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 E13 E14 E15 | 10.87 11.03 7.80 10.23 11.45 8.78 9.96 12.64 6.32 6.12 9.20 8.36 7.36 8.59 10.53 10.23 | 18.32 16.64 10.68 15.33 15.63 10.68 15.05 18.55 14.58 12.55 17.25 11.54 10.62 12.39 16.12 18.52 | 22.57 18.54 15.22 19.56 19.26 17.60 17.83 21.24 17.68 16.35 21.11 16.98 16.33 22.56 19.56 25.86 | 36.68 34.61 25.76 27.63 32.59 28.13 26.48 32.74 23.45 20.83 30.47 29.34 28.77 31.06 31.77 33.42 |
Conclusion: Exendin-4 small peptide simulating peptide (E1-E15) promotes that the activity and the native peptides of beta Cell of islet propagation are suitable, and tangible dose-effect relationship is arranged.
Experimental example 2
The mensuration of external anti-dipeptidyl peptidase enzyme stability
For measuring the vitro stability of each Exendin-4 small peptide simulating peptide, sample 37 ℃ of hydrolysis of carrying out dipeptidyl peptidase (DPPIV), is analyzed hydrolysate with HPLC then, and compare with Exendin-4.
Method: 37 ℃ of water-baths of dipeptidyl peptidase enzyme solution 30 minutes, add sample again, the final concentration that makes sample is 50 μ g/ml, 37 ℃ of water-bath hydrolysis, the 1 minute sample thief 400 μ l in every interval add acetic acid 400 μ l termination reactions.Sample is asked and is calculated the transformation period, result such as table 2 with the concentration that HPLC detects hydrolysate.
The stability of the external anti-DPPIV of table 2 Exendin-4 small peptide simulating peptide
| Group | Transformation period (min) |
| GLP-1 Exendin-4 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 E13 E14 E15 | 4.5 618 611 623 598 684 609 613 679 667 657 686 671 655 682 632 651 |
Conclusion: Exendin-4 small peptide simulating peptide (E1-E15) obviously is better than GLP-1 in the ability of external anti-dipeptidyl peptidase, and is suitable with Exendin-4.
Experimental example 3
Pharmacodynamic experiment
Laboratory animal: non-obese diabetic mouse (NOD mouse), body weight 17g+-2g, fasting 2 hours
Physiological saline group: intraperitoneal injection of saline
Exendin-4 group: abdominal injection Exendin-4
Experimental group E1-E15: abdominal injection Exendin-4 small peptide simulating peptide
The different time blood sample collection, the blood sugar detection test kit that uses Ministry of Health's Shanghai institute of Biological Products to produce is measured, and calculates glycemic index:
Experimental result such as table 3.
| Group | Dosage (μ g/kg) | Number of animals (only) | Blood sugar concentration (mmol/L) | ||||||||
| 0 | 10 | 15 | 45 | 60 | 90 | 180 | 270 | 480(min) | |||
| Physiological saline Exendin-4E1E2E3E4E5E6E7E8E9E10E11E12E13E14E15 | 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 | 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 | 17.86 13.65 14.42 11.13 14.50 12.77 9.91 14.99 16.50 10.96 11.82 12.55 11.34 13.78 12.63 11.76 13.46 | 18.85 14.87 15.76 11.88 16.63 13.46 10.12 16.52 17.45 11.53 13.00 14.46 12.57 14.68 13.74 13.15 15.02 | 18.54 12.99 11.56 6.58 6.03 6.68 5.90 13.44 11.32 7.08 9.21 9.75 8.76 11.89 7.13 8.96 11.86 | 17.65 10.56 10.07 7.90 9.11 7.71 6.74 10.95 8.63 6.54 6.86 8.76 6.04 9.87 8.06 7.14 10.75 | 18.44 10.99 10.94 9.96 9.25 11.85 7.88 12.44 9.95 7.23 8.94 9.36 7.23 1053 11.46 7.99 11.02 | 17.2311.2312.039.2910.708.347.8613.5410.268.869.6310.037.9611.649.128.1211.99 | 16.68 12.12 13.30 10.45 14.29 10.85 7.77 13.01 10.04 11.25 11.82 11.69 8.59 11.74 10.99 9.12 12.06 | 18.5212.8813.2410.4214.4210.817.7114.5211.4010.2312.3312.369.6812.5311.1210.0512.89 | 17.4613.2114.0010.3510.2011.749.3514.6511.5011.6512.4712.4510.3313.6511.9610.9813.06 |
Conclusion: the ability and the Exendin-4 of Exendin-4 small peptide simulating peptide (E1-E15) hypoglycemic are suitable.
Embodiment
The following examples can illustrate in greater detail the present invention, but do not limit the present invention in any form.
Embodiment 1
The solid phase synthesis of compd E 1 (Gln-Pro-Ser-Val-Gly-Met-Lys-Pro-Ser-Pro-Arg-His)
Raw material: Fmoc-Gln (Trt)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-His (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-His (Trt)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gln (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 2
The solid phase synthesis of compd E 2 (Ser-Val-Ser-Val-Gly-Met-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 3
The solid phase synthesis of compd E 3 (Ser-Val-Phe-Val-Gly-Met-Lys-Pro-Ser-Pro-Ser-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH, Fmoc-Phe-OH
, the linked reaction condensing agent is benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Ser (But)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Phe-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 4
The solid phase synthesis of compd E 4 (Ser-Val-Ser-Gly-Gly-Met-Lys-Pro-Ser-Pro-Arg-Lys)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Lys (Boc)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 5
The solid phase synthesis of compd E 5 (Ser-Val-Ser-Val-His-Met-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Met-OH, Fmoc-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-His (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-His (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 6
Compd E 6 (the solid phase synthesis of Gln-Pro-Ser-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-His)
Raw material: Fmoc-Gln (Trt)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-His (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-His (Trt)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-β-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gln (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 7
Compd E 7 (the solid phase synthesis of Ser-Val-Ser-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-oH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-8-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 8
Compd E 8 (the solid phase synthesis of Ser-Val-Phe-Val-Gly-Met-β-Lys-Pro-Ser-Pro-Ser-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-oH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-Phe-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Ser (But)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-β-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Phe-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 9
Compd E 9 (the solid phase synthesis of Ser-Val-Ser-Gly-Gly-Met-β-Lys-Pro-Ser-Pro-Arg-Lys)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Lys (Boc)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-β-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 10
Compd E 10 (the solid phase synthesis of Ser-Val-Ser-Val-His-Met-β-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Met-OH, Fmoc-β-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-His (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-β-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-His (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 11
Compd E 11 (the solid phase synthesis of Gln-Pro-Ser-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-His)
Raw material: Fmoc-Gln (Trt)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-γ-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-His (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-His (Trt)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-γ-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gln (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 12
Compd E 12 (the solid phase synthesis of Ser-Val-Ser-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-γ-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-γ-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 13
Compd E 13 (the solid phase synthesis of Ser-Val-Phe-Val-Gly-Met-γ-Lys-Pro-Ser-Pro-Ser-Pro)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-γ-Lys (Boc)-OH, Fmoc-Phe-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Ser (But)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-γ-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Phe-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 14
Compd E 14 (the solid phase synthesis of Ser-Val-Ser-Gly-Gly-Met-γ-Lys-Pro-Ser-Pro-Arg-Lys)
Raw material: Fmoc-Ser (But)-OH, Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-Met-OH, Fmoc-γ-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Lys (Boc)-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-γ-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Gly-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 15
Compd E 15 (the solid phase synthesis of Ser-Val-Ser-Val-His-Met-γ-Lys-Pro-Ser-Pro-Arg-Pro)
Raw material: Fmoc-Pro-OH, Fmoc-Ser (But)-OH, Fmoc-Val-OH, Fmoc-Met-OH, Fmoc-γ-Lys (Boc)-OH, Fmoc-Arg (pbf)-OH, Fmoc-Hi s (Trt)-OH, linked reaction condensing agent are benzene a pair of horses going side by side triazole-1-oxygen-three (dimethylamino) phosphorus hexafluorophosphate (BOP), 1-hydroxyl benzotriazole (HOBT), N-methylmorpholine (NMM).Deprotection agent is 20% piperidines/DMF solution.Cutting reagent is a trifluoroacetic acid.
Reaction: take by weighing Rink Amide mbha resin, resin room temperature swelling 30 minutes in DMF is with DMF washing three times.Add 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction.With DMF washing three times, add Fmoc-Pro-OH again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution, 1 hour deprotection of room temperature reaction with DMF washing three times.With DMF washing three times, add Fmoc-Arg (pbf)-OH more again, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Pro-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-γ-Lys (Boc)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Met-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-His (Trt)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Val-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.Add Fmoc-Ser (But)-OH, room temperature reaction 3 hours, the back adds 20% piperidines/DMF solution with DMF washing three times, and 1 hour deprotection of room temperature reaction is again with DMF washing three times.
Cutting: add the cutting reagent trifluoroacetic acid behind the resin drying of above-mentioned reaction, room temperature reaction 2 hours filters, and filtrate is precipitated in ether, and drying obtains thick product.
Purifying: crude product is soluble in water, use C
18Reversed-phase column carries out purifying, and elutriant is that A is the 0.1%TFA aqueous solution mutually, and B is acetonitrile mutually, and flow velocity is 20ml/min, and the detection wavelength is 214nm, collects main peak, and freeze-drying obtains pure product.
Embodiment 16
The method that takes by weighing embodiment 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15 respectively obtains each 100mg of sample of E1-E15, be dissolved in 1000ml 0.9% sodium chloride solution, after mixing, be distributed into the injection liquid that 0.1mg/ml/ props up concentration and seal in medicine bottle, product is made in sterilization.
Embodiment 17
The method that takes by weighing embodiment 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15 respectively obtains each 100mg of sample of E1-E15, be dissolved in the 1000ml water and make the aqueous solution, after mixing, being distributed into the injection liquid that 0.1mg/ml/ props up concentration seals in medicine bottle, sterilization, finished product is made in freeze-drying.
Embodiment 18
The method that takes by weighing embodiment 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15 respectively obtains each 100g of sample of E1-E15, and medical starch 0.5kg makes capsule by known capsule technology of preparing and equipment, every 10mg.Other project should meet requirement under Pharmacopoeia of People's Republic of China version capsule in 2000 item.
Embodiment 19
The method that takes by weighing embodiment 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15 respectively obtains each 100g of sample of E1-E15, Microcrystalline Cellulose 560g, lactose hydrous 380g, Magnesium Stearate 200g, silicon oxide 30g, make tablet by known tabletting technology and equipment, every 10mg.Other project should meet requirement under Pharmacopoeia of People's Republic of China version tablet in 2000 item.
Claims (8)
1. Exendin-4 small peptide simulating peptide, it is characterized in that: the structure of peptide is X-Met-Lys-Pro-Ser-Pro-Y, and wherein, X is Gln-Pro-Ser-Val-Gly, and Y is Arg-His; Or X is Ser-Val-Phe-Val-Gly, and Y is Ser-Pro; Or X is Ser-Val-Ser-Gly-Gly, and Y is Arg-Lys; Or X is Ser-Val-Ser-Val-His, and Y is Arg-Pro; Or X is Ser-Val-Ser-Val-Gly, and Y is Arg-Pro, and the Lys among the Met-Lys-Pro-Ser-Pro is that β or γ replace.
2. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the X in the compound is Gln-Pro-Ser-Val-Gly, and Y is Arg-His.
3. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the X in the compound is Ser-Val-Phe-Val-Gly, and Y is Ser-Pro.
4. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the X in the compound is Ser-Val-Ser-Gly-Gly, and Y is Arg-Lys.
5. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the X in the compound is Ser-Val-Ser-Val-His, and Y is Arg-Pro.
6. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the Lys of the Met-Lys-Pro-Ser-Pro in the compound is that β or γ replace.
7. Exendin-4 small peptide simulating peptide according to claim 1 is characterized in that: the X in the compound is Ser-Val-Ser-Val-Gly, and Y is Arg-Pro, and the Lys among the Met-Lys-Pro-Ser-Pro is that β or γ replace.
8. the application of the described Exendin-4 simulating peptide of claim 1 in the preparation Remedies for diabetes.
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| CN106554403B (en) | 2015-09-25 | 2021-08-31 | 博瑞生物医药（苏州）股份有限公司 | Exenatide modifier and application thereof |
| IL270199B1 (en) * | 2017-05-04 | 2024-03-01 | Follicum Ab | Peptides for treatment of diabetes |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6498027B1 (en) * | 1998-05-20 | 2002-12-24 | Introgene B.V. | Targeted delivery through a cationic amino acid transporter |
| CN1162446C (en) * | 2001-05-10 | 2004-08-18 | 上海华谊生物技术有限公司 | Insulinotropic hormone secretion peptide derivative |
| CN1927879A (en) * | 2006-09-25 | 2007-03-14 | 吉林大学 | Thymopentapeptide active isomer and application thereof in pharmaceutical preparation |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6498027B1 (en) * | 1998-05-20 | 2002-12-24 | Introgene B.V. | Targeted delivery through a cationic amino acid transporter |
| CN1162446C (en) * | 2001-05-10 | 2004-08-18 | 上海华谊生物技术有限公司 | Insulinotropic hormone secretion peptide derivative |
| CN1927879A (en) * | 2006-09-25 | 2007-03-14 | 吉林大学 | Thymopentapeptide active isomer and application thereof in pharmaceutical preparation |
Non-Patent Citations (2)
| Title |
|---|
| 葛晓宇等.Exendin-4及其类似物的研究近况.药学进展31 9.2007,31(9),403-407. |
| 葛晓宇等.Exendin-4及其类似物的研究近况.药学进展31 9.2007,31(9),403-407. * |
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