CN116284218A - Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance - Google Patents

Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance Download PDF

Info

Publication number
CN116284218A
CN116284218A CN202310264615.4A CN202310264615A CN116284218A CN 116284218 A CN116284218 A CN 116284218A CN 202310264615 A CN202310264615 A CN 202310264615A CN 116284218 A CN116284218 A CN 116284218A
Authority
CN
China
Prior art keywords
compound
compounds
ext
tag
dichloromethane
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310264615.4A
Other languages
Chinese (zh)
Inventor
徐华栋
李贤伟
沈美华
李亮
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Changzhou University
Original Assignee
Changzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Changzhou University filed Critical Changzhou University
Priority to CN202310264615.4A priority Critical patent/CN116284218A/en
Publication of CN116284218A publication Critical patent/CN116284218A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
    • C07C41/01—Preparation of ethers
    • C07C41/18—Preparation of ethers by reactions not forming ether-oxygen bonds
    • C07C41/26—Preparation of ethers by reactions not forming ether-oxygen bonds by introduction of hydroxy or O-metal groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/61—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds by reactions not involving the formation of >C = O groups
    • C07C45/64—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds by reactions not involving the formation of >C = O groups by introduction of functional groups containing oxygen only in singly bound form
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention belongs to the technical field of medicine synthesis, and particularly relates to a method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance. Firstly, synthesizing four liquid phase carriers TAG-A, TAG-B, TAG-C and TAG-D, then condensing the four liquid phase carriers with Fmoc-Lys (Boc) -OH to obtain compounds 4A-4D, and deprotecting to obtain compounds 5A-5D; condensing the compound 5A-5D with Fmoc-His (Trt) -OH to obtain a compound 6A-6D, deprotecting to obtain a compound 7A-7D, condensing the compound 7A-7D with Fmoc-protected glycine to obtain a compound 8A-8D, deprotecting to obtain a compound 9A-9D, condensing the compound 9A-9D with palmitic acid to obtain a full-protected peptide compound 10A-10D; finally, the palmitoyl tripeptide-1 compound can be obtained by trifluoroacetic acid cleavage.

Description

Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance
Technical Field
The invention belongs to the field of medicine synthesis, and particularly relates to a method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance.
Background
Palmitoyl Tripeptide-1 (Pal-Tripeptide-1) is composed of three amino acids and one fatty acid, and has an amino acid sequence of glycine-histidine-lysine (GHK), and palmitic acid can be connected with the N-terminal of GHK to form a target peptide after acylation treatment. Palmitoyl tripeptide-1 is a special amino acid polypeptide, acts on dermis layers, can enable skin to be more compact, enable wrinkles to be relaxed, promote metabolism of the skin, stimulate proliferation of cells, and can achieve the effects of removing wrinkles, whitening, resisting aging and increasing skin luster.
The synthesis method of palmitoyl tripeptide-l comprises two methods of solid-phase polypeptide synthesis and liquid-phase polypeptide synthesis, but the simple solid-phase method for synthesizing palmitoyl tripeptide-l has higher cost due to shorter peptide sequence, expensive solid-phase carrier and large amino acid consumption; the liquid phase synthesis method has the advantages of small amino acid consumption and high yield, but the existing liquid phase synthesis method has complex post-treatment process, uses a large amount of organic solvents, has high cost and is not environment-friendly.
CN108218956a discloses a liquid phase synthesis method of palmitoyl tripeptide-1. The method comprises the steps of synthesizing palmitoyl tripeptide-1 in a segmented manner, respectively synthesizing a dipeptide fragment H-His-Lys (Z) -OBzl and another fragment Pal-Gly-OH in the palmitoyl tripeptide-1 sequence, then synthesizing the full-protection peptide of palmitoyl tripeptide by using the two fragments in an activated ester method, and obtaining the final product palmitoyl tripeptide-1 through hydrogenation reaction. During the synthesis of Pal-Gly-OH, the Pal-OH residue is difficult to quantitatively detect, the synthesis of the fragment needs to calculate the feeding amount according to the synthesis condition of another fragment, meanwhile, the problem that the intermediate Pal-Gly-ONb has high activity and is difficult to preserve also leads to difficult mass production, and the final step of hydrogenation is easy to have explosion risk and is not suitable for mass production.
In the liquid phase synthesis of the disclosed palmitoyl tripeptide-1, both CN112409444A and CN114891063A react with Boc-Lys (Z) -OH and benzyl bromide to obtain Boc-Lys (Z) -OBzl, and then remove Boc to obtain H-Lys (Z) -OBzl; condensing with Boc-His-OH, and removing Boc to obtain H-His-Lys (Z) -OBzl; repeating the amino acid coupling step, and sequentially coupling Boc-Gly-OH and Pal-OH according to the peptide sequence to obtain Pal-Gly-His-Lys (z) -OBzl; finally hydrogenation debenzylation is carried out to obtain Pal-Gly-His-Lys-OH. The method is not beneficial to large-scale production due to the fact that various controlled reagents are used, palladium carbon used in the last step of hydrogenation reaction is dangerous, fire and explosion risks are prone to occur, and the method is not suitable for large-scale production.
The synthesis of palmitoyl tripeptide-1 is mostly carried out by adopting a strategy of protecting amino by Boc, wherein Boc-Lys (Z) -OBzl is firstly obtained and then condensed with Boc-His-OH and Pal-Gly dipeptide or glycine is firstly connected and then palmitic acid is firstly connected to obtain the full-protection peptide of palmitoyl tripeptide-1, and finally hydrogenation debenzylation is carried out to obtain the palmitoyl tripeptide-1. However, in the existing synthetic route, it is not difficult to find that the post-treatment purification process is complicated, the types of the used solvents are more, the solvent amount is larger, and some solvents belong to the controlled solvents, so that the mass production is not facilitated. Palladium on carbon and hydrogen are used in the final deprotection of the fully protected peptide, which also presents a safety hazard in production with the risk of explosion and ignition.
The liquid phase carrier technology developed in recent years combines the advantages of the solid phase polypeptide synthesis method and the traditional liquid phase polypeptide synthesis method, and the C end of the target polypeptide is additionally provided with a protecting group carrier with a long hydrophobic alkyl chain, each step of reaction can be carried out in halogenated solvent, and the reaction intermediate can be precipitated by using polar organic solvent. Compared with the traditional liquid-phase polypeptide synthesis method, the post-treatment operation is simple, and the purity of the intermediate in each step is relatively high; compared with the solid-phase polypeptide synthesis method, the method improves the utilization rate of amino acid and saves the cost. However, the existing liquid phase carrier assists in the synthesis of polypeptide drugs: (1) the method comprises the following steps The product may be slightly viscous during the deprotection step, (2): the product may have a reduced yield during settling in some steps due to the finer product particles and the presence of leaks during the filtration operation.
Disclosure of Invention
The invention aims at overcoming the defects of the prior art and provides a method for synthesizing palmitoyl tripeptide-1 based on the assistance of a liquid carrier.
Ext> theext> inventionext> firstlyext> usesext> 4ext>,ext> 4ext> -ext> dihydroxybenzophenoneext> (ext> compoundext> 1ext>)ext> asext> aext> rawext> materialext>,ext> andext> theext> compoundext> 2ext> isext> obtainedext> byext> theext> reactionext> ofext> alkylationext> reactionext> andext> 1ext> -ext> bromooctadecaneext>,ext> andext> thenext> theext> compoundext> 2ext> isext> reducedext> byext> sodiumext> borohydrideext> toext> obtainext> aext> hydrophobicext> liquidext> phaseext> carrierext> compoundext> 3ext> (ext> TAGext> -ext> Aext>)ext>.ext>
Wherein the dosage of the 1-bromooctadecane is between 2.2 and 3 equivalents; the sodium borohydride is used in an amount between 1.0 equivalent and 3.0 equivalents.
Figure BDA0004132695210000031
1 liquid phase Carrier Synthesis route one
Figure BDA0004132695210000032
2 liquid phase carrier synthesis route II
The synthesis of palmitoyl tripeptide-1 may also be performed using compound 14, compound 17, or compound 19 as a liquid phase carrier.
The compound 12 is first alkylated to give the compound 13, and the compound 13 is then reduced by sodium borohydride to give the compound 14 (TAG-B).
Compound 14 is reacted with thionyl chloride to give compound 15, compound 15 is reacted with SN 2 The reaction gave compound 16, compound 16 was then reduced with sodium borohydride to give compound 17 (TAG-C).
In addition, compound 15 can also be prepared by SN 2 The reaction gave compound 18, and compound 18 was then reduced with sodium borohydride to give compound 19 (TAG-D).
The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier comprises the following steps: condensing liquid phase carrier TAG-A, TAG-B, TAG-C or TAG-D (compound 3/compound 14/compound 17/compound 19) with Fmoc-Lys (Boc) -OH to obtain Fmoc-Lys (Boc) -TAG (compound)4A-4D), deprotection to give NH 2 -Lys (Boc) -TAG (compounds 5A-5D); condensing with Fmoc-His (Trt) -OH to obtain Fmoc-His (Trt) -Lys (Boc) -TAG (compounds 6A-6D), and deprotecting to obtain NH 2 -His (Trt) -Lys (Boc) -TAG (Compounds 7A-7D), condensing with Fmoc-protected glycine to give Fmoc-Gly-His (Trt) -Lys (Boc) -TAG (Compounds 8A-8D), deprotecting to give NH 2 -Gly-His (Trt) -Lys (Boc) -TAG (compounds 9A-9D), and condensing with palmitic acid to obtain full-protection peptide Pal-Gly-His (Trt) -Lys (Boc) -TAG (compounds 10A-10D). Finally, the palmitoyl tripeptide-1 (compound 11) can be obtained by trifluoroacetic acid cleavage.
Figure BDA0004132695210000041
Palmitoyl tripeptide-1 synthesis route
Further: the synthesis method of the compounds 4A-4D comprises the following steps: dichloromethane, a liquid carrier (TAG-OH) (compound 3/compound 14/compound 17/compound 19), fmoc-Lys (Boc) -OH (1.1 equivalent to 1.8 equivalents), N' -diisopropylcarbodiimide (1.1 equivalent to 1.8 equivalents), 4-dimethylaminopyridine (0.1 equivalent to 0.6 equivalents) were added to the flask at a time, stirred for 3 hours, then dichloromethane was concentrated, methanol was added under ice bath to precipitate a solid, and stirring was continued for 10 minutes, and the solid powder compounds 4A to 4D were collected by filtration.
The synthesis method of the compounds 5A-5D comprises the following steps: dichloromethane, the compounds 4A-4D and 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.0 equivalent-2.5 equivalent) are sequentially added into the flask, stirred for 30 minutes at room temperature, then the dichloromethane is concentrated, acetonitrile is added under ice bath, stirring is continued for 10 minutes after solid precipitation, and then the solid powder compounds 5A-5D are filtered and collected.
The synthesis method of the compounds 6A-6D comprises the following steps: dichloromethane, fmoc-His (Trt) -OH (1.1-1.8 equivalents), N-diisopropylethylamine (1.1-1.8 equivalents), N' -diisopropylcarbodiimide (1.1-1.8 equivalents) and 1-hydroxybenzotriazole (1.1-1.8 equivalents) are sequentially added into a round bottom flask, stirring is carried out for 10 minutes, then the compounds 5A-5D are added into a reaction liquid for continuous stirring for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, then filtration is carried out, and solid powder 6A-6D is collected.
The synthesis method of the compounds 7A-7D comprises the following steps: dichloromethane, compounds 6A-6D and 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.0 equivalent-2.5 equivalent) were added to the round bottom flask in sequence, stirred at room temperature for 30 minutes, then the dichloromethane was concentrated, acetonitrile was added under ice bath to precipitate solids, and then stirring was continued for 10 minutes, and then filtration was carried out to collect solid powder compounds 7A-7D.
The synthesis method of the compounds 8A-8D comprises the following steps: dichloromethane, fmoc-Gly-OH (1.1-1.8 equivalent), N-diisopropylethylamine (1.1-1.8 equivalent), N' -diisopropylcarbodiimide (1.1-1.8 equivalent) and 1-hydroxybenzotriazole (1.1-1.8 equivalent) are sequentially added into a round bottom flask, stirred for 10 minutes, then the compound 7A-7D is added into the reaction liquid, stirring is continued for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, solid is separated out, stirring is continued for 10 minutes, then filtration is carried out, and the solid powder compound 8A-8D is collected.
The synthesis method of the compounds 9A-9D comprises the following steps: dichloromethane, the compounds 8A-8D and 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.0 equivalent-2.5 equivalent) are sequentially added into a round bottom flask, stirred for 30 minutes at room temperature, then the dichloromethane is concentrated, acetonitrile is added under ice bath to precipitate solids, stirring is continued for 10 minutes, and then the solid powder compounds 9A-9D are filtered and collected.
The synthesis method of the compounds 10A-10D comprises the following steps: dichloromethane, palmitic acid (1.1-1.8 equivalent), N-diisopropylethylamine (1.1-1.8 equivalent), N' -diisocarbodiimides (1.1-1.8 equivalent) and 1-hydroxybenzotriazole are sequentially added into a round bottom flask, stirring is carried out for 10 minutes, then the compounds 9A-9D are added into a reaction liquid for continuous stirring for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, filtering is carried out, solid powder is collected, and the solid powder is dried in a vacuum drying oven at 30 ℃ to obtain white solid compounds 10A-10D.
The synthesis method of palmitoyl tripeptide-1 comprises the following steps: adding dichloromethane, compounds 10A-10D and cutting fluid into a round bottom flask in turn, continuously stirring for 3 hours after the reaction fluid is dissolved, concentrating the reaction fluid, continuously stirring for 30 minutes after isopropyl ether is added at 0 ℃ to precipitate solids, filtering, collecting a filter cake, stirring and dissolving the filter cake with 100 milliliters of water, filtering, preparing and purifying the filtrate, and freeze-drying the collected fluid by a freeze dryer to obtain a white solid final product palmitoyl tripeptide-1.
The beneficial effects are that:
the palmitoyl tripeptide-1 is synthesized by using the liquid phase carrier, the reaction condition is mild by using methylene dichloride as a solvent, the post-treatment process is simple, and the product can be precipitated out by using a large polar solvent methanol or acetonitrile after the reaction is finished, so that the palmitoyl tripeptide-1 is suitable for large-scale production.
The carrier has solubility, and the reaction is homogeneous phase reaction; improves the utilization rate of amino acid and saves the cost.
Description of the drawings:
fig. 1: ms plot of palmitoyl tripeptide-1.
Detailed Description
The present invention will be further explained below with reference to specific embodiments for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, but is not intended to limit the scope of the present invention.
Example 1
Figure BDA0004132695210000071
150 ml of N, N-dimethylformamide, compound 1 (10 g, 46.68 mmol), potassium carbonate (14.2 g, 102.7 mmol) and 1-bromooctadecane (34.4 g, 102.7 mmol) were successively added to a 500 ml round bottom flask at 25.+ -. 5 ℃ and then reacted at 90 ℃ for 4 hours. After the reaction, the mixture was poured into 750 ml of water while it was still hot, stirred at 25.+ -. 5 ℃ for 30 minutes, then filtered, and the filter cake was slurried with 500 ml of methanol and then filtered, and the white solid was collected and dried in a forced air drying oven at 45 ℃ to give 2.6 g of a white solid compound with a yield of 97%.
Figure BDA0004132695210000072
350 ml of tetrahydrofuran, compound 2 and 35 ml of methanol are added into a 1 liter round bottom flask in sequence at 25+/-5 ℃ and then heated to 60 ℃, and sodium borohydride (1.7 g, 45 mmol) is added to react for 2 hours after the reaction solution is dissolved. Ext> afterext> theext> reactionext> isext> finishedext>,ext> afterext> theext> reactionext> solutionext> isext> cooledext> toext> roomext> temperatureext>,ext> waterext> isext> slowlyext> addedext> intoext> theext> reactionext> solutionext>,ext> whenext> noext> obviousext> bubbleext> isext> generatedext> inext> theext> reactionext> solutionext>,ext> theext> reactionext> solutionext> isext> pouredext> intoext> 2ext> litersext> ofext> waterext>,ext> stirredext> forext> 30ext> minutesext> andext> thenext> filteredext>,ext> aext> filterext> cakeext> isext> pulpedext> byext> 500ext> millilitersext> ofext> methanolext> andext> thenext> filteredext>,ext> andext> aext> whiteext> solidext> isext> collectedext> andext> driedext> atext> 45ext> ℃ext> inext> aext> forcedext> airext> dryingext> ovenext>,ext> soext> thatext> 31.3ext> gext> ofext> aext> whiteext> solidext> compoundext> 3ext> (ext> TAGext> -ext> Aext>)ext> isext> obtainedext>,ext> andext> theext> yieldext> isext> 96ext>%ext>.ext>
Figure BDA0004132695210000081
Ext> 50ext> mlext> ofext> methyleneext> chlorideext>,ext> compoundext> 3ext> (ext> TAGext> -ext> Aext>,ext> 5ext> gext>,ext> 6.93ext> mmolext>)ext>,ext> Fmocext> -ext> Lysext> (ext> Bocext>)ext> -ext> OHext> (ext> commerciallyext> availableext> asext> Duext> Zhengext> Yuanext>)ext> (ext> 3.9ext> gext>,ext> 8.32ext> mmolext>)ext>,ext> Next>'ext> -ext> diisopropylcarbodiimideext> (ext> 1.6ext> mlext>,ext> 10.4ext> mmolext>)ext> andext> 4ext> -ext> dimethylaminopyridineext> (ext> 169ext> mgext>,ext> 1.387ext> mmolext>)ext> wereext> addedext> inext> thisext> orderext> toext> aext> 250ext> mlext> roundext> bottomext> flaskext> andext> stirredext> forext> 3ext> hoursext>,ext> afterext> whichext> theext> methyleneext> chlorideext> wasext> concentratedext> toext> aboutext> 5ext> mlext>,ext> andext> 100ext> mlext> ofext> methanolext> wasext> addedext> underext> iceext> bathext> toext> precipitateext> aext> solidext> andext> stirredext> forext> 10ext> minutesext>.ext> Then, the solid powdery compound 4A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000082
50 ml of dichloromethane, compound 4A and 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.036 ml, 6.93 mmol) were added sequentially to a 250 ml round bottom flask at 25+ -5 ℃ and stirred at room temperature for 30 minutes, then the dichloromethane was concentrated to about 5 ml, and 100 ml of acetonitrile was added under ice bath to precipitate solids, and stirring was continued for 10 minutes. Then, the solid powdery compound 5A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000091
50 ml of dichloromethane, fmoc-His (Trt) -OH (5.157 g, 8.32 mmol) (commercially available as Zheng Yuan), N-diisopropylethylamine (1.56 ml, 9 mmol), N' -diisopropylcarbodiimide (1.6 ml, 10.4 mmol) and 1-hydroxybenzotriazole (1.4 g, 10.4 mmol) were added sequentially to a 250 ml round bottom flask at 15.+ -. 5 ℃ and stirred for 10 minutes, then compound 5A was added to the reaction mixture and stirred for 3 hours. Then, methylene chloride was concentrated to about 5 ml, and 100 ml of methanol was added to the mixture in an ice bath to precipitate a solid, followed by stirring for 10 minutes. Then, the solid powdery compound 6A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000092
50 ml of dichloromethane, 6A of compound, 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.036 ml, 6.93 mmol) were added in sequence to a 250 ml round bottom flask at 25+ -5 ℃ and stirred at room temperature for 30 minutes, then the dichloromethane was concentrated to about 5 ml, and 100 ml of acetonitrile was added under ice bath to precipitate solids, and stirring was continued for 10 minutes. Then, the solid powdery compound 7A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000101
75 ml of methylene chloride, fmoc-Gly-OH (2.47 g, 8.32 mmol), N-diisopropylethylamine (1.56 ml, 9 mmol), N' -diisopropylcarbodiimide (1.6 ml, 10.4 mmol), 1-hydroxybenzotriazole (1.4 g, 10.4 mmol) were added in this order to a 250 ml round bottom flask at 20.+ -. 5 ℃ and stirred for 10 minutes, then compound 7A was added to the reaction mixture and stirred for 3 hours. Then, methylene chloride was concentrated to about 5 ml, and 100 ml of methanol was added to the mixture in an ice bath to precipitate a solid, followed by stirring for 10 minutes. Then, the solid powdery compound 8A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000102
50 ml of dichloromethane, compound 8A, 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.036 ml, 6.93 mmol) were added sequentially to a 250 ml round bottom flask at 25+ -5deg.C, stirred at room temperature for 30 min, then the dichloromethane was concentrated to about 5 ml, and 100 ml of acetonitrile was added under ice bath to precipitate solids, followed by stirring for 10 min. Then, the solid powdery compound 9A was collected by filtration, and the next step was taken out by suction.
Figure BDA0004132695210000103
75 ml of methylene chloride, palmitic acid (2.13 g, 8.32 mmol), N-diisopropylethylamine (1.56 ml, 9 mmol), N' -diiso-carbodiimides (1.6 ml, 10.4 mmol), 1-hydroxybenzotriazole, (1.4 g, 10.4 mmol) were added in this order to a 250 ml round bottom flask at 20.+ -. 5 ℃ and stirred for 10 minutes, then compound 9A was added to the reaction mixture and stirred for 3 hours. Then, methylene chloride was concentrated to about 5 ml, and 100 ml of methanol was added to the mixture in an ice bath to precipitate a solid, followed by stirring for 10 minutes. Then, the mixture was filtered, and the solid powder was collected and dried in a vacuum oven at 30℃to give 10.55 g of a white solid compound 10A in a yield of 93.8%.
Figure BDA0004132695210000111
50 ml of dichloromethane, compound 10A and 50 ml of cutting fluid (trifluoroacetic acid: triisopropylsilane: water=95:2.5:2.5) are added in sequence into a 500 ml round bottom flask at 10+ -5 ℃ and stirring is continued for 3 hours after the reaction solution is dissolved. The reaction mixture was concentrated to about 20 ml, and 200 ml of isopropyl ether was added at 0℃to precipitate a solid, followed by stirring for 30 minutes. Then filtering, collecting a filter cake, stirring and dissolving the filter cake with 100 milliliters of water, filtering, and preparing and purifying the filtrate. The collected liquid is freeze-dried by a freeze dryer to obtain a white solid final product, the yield is 62.5%, and the purity is more than 98%.
Example 2
Figure BDA0004132695210000112
150 ml of N, N-dimethylformamide, compound 12 (10 g, 72.46 mmol), potassium carbonate (30 g, 217 mmol), 1-bromooctadecane (53.14 g, 159 mmol) were added sequentially to a 500 ml round bottom flask at 25.+ -. 5 ℃ and then reacted at 90 ℃ for 4 hours. After the reaction, the mixture was poured into 750 ml of water while it was still hot, stirred at 25.+ -. 5 ℃ for 30 minutes, then filtered, and the filter cake was slurried with 500 ml of methanol and then filtered, and the white solid was collected and dried in a forced air drying oven at 45 ℃ to give 13.57 g of a white solid compound with a yield of 98%.
Figure BDA0004132695210000121
400 ml of tetrahydrofuran, 13 of compound and 45 ml of methanol are added into a 1 liter round bottom flask in sequence at 25+/-5 ℃ and then heated to 60 ℃, and sodium borohydride (2.68 g, 71 mmol) is added to react for 2 hours after the reaction solution is dissolved. After the reaction is finished, slowly dripping water into the reaction liquid after the reaction liquid is cooled to room temperature, pouring the reaction liquid into 2L of water when no obvious bubble is generated in the reaction liquid, stirring for 30 minutes, filtering, pulping a filter cake with 500 ml of methanol, filtering, collecting white solid, and drying at 45 ℃ in a forced air drying oven to obtain 45 g of white solid compound 14 (TAG-B), wherein the yield is 98%.
The present invention is based on an example of a liquid phase carrier-assisted synthesis method of palmitoyl tripeptide-1, and the procedure is the same as in example 1 except that a liquid phase carrier (compound 14 (TAG-B) is used in this example) is used differently from example 1. The yield of palmitoyl tripeptide-1 was 53%.
Example 3
Figure BDA0004132695210000122
100 ml of dichloromethane, 14 of compound and 1 ml of thionyl chloride are sequentially added into a 250 ml round-bottom flask at 0+/-5 ℃ to react for 1 hour at 0 ℃, after the reaction is finished, the reaction solution is concentrated to dryness, 100 ml of dichloromethane is added to wash once with 5% sodium bicarbonate aqueous solution, and then the dichloromethane is dried with anhydrous magnesium sulfate and concentrated to dryness to obtain 15.8 g of product compound with 93% yield.
Figure BDA0004132695210000131
80 ml of N, N-dimethylformamide, compound 15 (4.8 g, 7.2 mmol), potassium carbonate (2 g, 14.5 mmol) and p-hydroxybenzaldehyde (1.06 g, 8.7 mmol) were successively added to a 500 ml round bottom flask at 25.+ -. 5 ℃ and then reacted at 80 ℃ for 4 hours. After the reaction, the mixture was poured into 500 ml of water while it was still hot, stirred at 25.+ -. 5 ℃ for 30 minutes, then filtered, and the filter cake was slurried with 200 ml of methanol and then filtered, and the white solid was collected and dried in a forced air drying oven at 45 ℃ to give 16.1 g of a white solid compound with a yield of 94%.
Figure BDA0004132695210000132
45 ml of tetrahydrofuran, 16 of compound and 5 ml of methanol are sequentially added into a 250 ml round bottom flask at 25+/-5 ℃ and then heated to 60 ℃, and sodium borohydride (0.26 g, 6.8 mmol) is added to react for 2 hours after the reaction solution is dissolved. After the reaction is finished, after the reaction solution is cooled to room temperature, water is slowly added dropwise into the reaction solution, when no obvious bubble is generated in the reaction solution, the reaction solution is poured into 300 ml of water, stirred for 30 minutes and then filtered, a filter cake is pulped by 100 ml of methanol and then filtered, and a white solid is collected and dried at 45 ℃ in a forced air drying oven, so that 4.9 g of white solid compound 17 (TAG-C) is obtained, and the yield is 96%.
The present invention is based on one example of a liquid phase carrier-assisted synthesis of palmitoyl tripeptide-1, and the procedure is identical to that of example 1, except that the liquid phase carrier used is different from that of example 1, which uses compound 17 (TAG-C). The yield of palmitoyl tripeptide-1 was 65% as a result.
Example 4
Figure BDA0004132695210000141
80 ml of N, N-dimethylformamide, compound 15 (4.8 g, 7.2 mmol), potassium carbonate (2 g, 14.5 mmol), 4- (allyloxy) -2-hydroxybenzaldehyde (1.54 g, 8.7 mmol) were successively added to a 500 ml round bottom flask at 25.+ -. 5 ℃ and then reacted at 80 ℃ for 4 hours. After the reaction, the mixture was poured into 500 ml of water while it was still hot, stirred at 25.+ -. 5 ℃ for 30 minutes, then filtered, and the filter cake was slurried with 200 ml of methanol, then filtered, and the white solid was collected and dried in a forced air drying oven at 45 ℃ to give 18.3 g of a white solid compound with a yield of 91%.
Figure BDA0004132695210000142
45 ml of tetrahydrofuran, 18 of compound and 5 ml of methanol are sequentially added into a 250 ml round bottom flask at 25+/-5 ℃ and then heated to 60 ℃, and sodium borohydride (0.25 g, 6.6 mmol) is added to react for 2 hours after the reaction solution is dissolved. After the reaction is finished, after the reaction solution is cooled to room temperature, water is slowly added dropwise into the reaction solution, when no obvious bubble is generated in the reaction solution, the reaction solution is poured into 300 ml of water, stirred for 30 minutes and then filtered, a filter cake is pulped by 100 ml of methanol and then filtered, and a white solid is collected and dried at 45 ℃ in a forced air drying oven, so that 4.7 g of white solid compound 19 (TAG-D) is obtained, and the yield is 88%.
The procedure of example 2 was identical except that the liquid phase carrier used was different from example 2 (this example uses compound 19 (TAG-D)) based on one example of a liquid phase carrier-assisted synthesis method for palmitoyl tripeptide-1. The yield of palmitoyl tripeptide-1 was finally found to be 63%.
The above embodiments are only for illustrating the technical solution of the present invention, and the description thereof is more specific and detailed, but not limiting. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention.

Claims (10)

1. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier is characterized by comprising the following steps of: condensing four liquid phase carriers TAG-A, TAG-B, TAG-C or TAG-D with Fmoc-Lys (Boc) -OH respectively to obtain compounds 4A-4D, and deprotecting to obtain compounds 5A-5D; condensing the compound 5A-5D with Fmoc-His (Trt) -OH to obtain a compound 6A-6D, deprotecting to obtain a compound 7A-7D, condensing the compound 7A-7D with Fmoc-protected glycine to obtain a compound 8A-8D, deprotecting to obtain a compound 9A-9D, condensing the compound 9A-9D with palmitic acid to obtain a full-protected peptide compound 10A-10D; finally obtaining palmitoyl tripeptide-1 compound 11 through trifluoroacetic acid cleavage; the specific formula is as follows:
Figure FDA0004132695200000011
2. ext> theext> methodext> forext> synthesizingext> palmitoylext> tripeptideext> -ext> 1ext> basedext> onext> theext> liquidext> phaseext> carrierext> assistanceext> accordingext> toext> claimext> 1ext>,ext> whereinext> theext> synthesisext> methodext> ofext> TAGext> -ext> Aext> isext> asext> followsext>:ext> Ext> 4ext>,ext> 4ext> -ext> dihydroxybenzophenoneext> isext> usedext> asext> aext> rawext> materialext>,ext> theext> rawext> materialext> isext> reactedext> withext> 1ext> -ext> bromooctadecaneext> throughext> alkylationext> reactionext> toext> obtainext> aext> compoundext> 2ext>,ext> andext> theext> compoundext> 2ext> isext> reducedext> byext> sodiumext> borohydrideext> toext> obtainext> aext> hydrophobicext> liquidext> phaseext> carrierext> TAGext> -ext> Aext>,ext> whereinext> theext> specificext> formulaext> isext> asext> followsext>:ext>
Figure FDA0004132695200000012
The synthesis method of TAG-B comprises the following steps: the compound 12 is subjected to alkylation reaction to obtain a compound 13, and the compound 13 is reduced by sodium borohydride to obtain a liquid carrier TAG-B;
the synthesis method of TAG-C comprises the following steps: TAG-B under the action of thionyl chloride to obtain a compound 15, wherein the compound 15 is prepared by SN 2 ReactionObtaining a compound 16, and reducing the compound 16 by sodium borohydride to obtain a liquid carrier TAG-C;
the synthesis method of TAG-D comprises the following steps: TAG-B under the action of thionyl chloride to obtain a compound 15, wherein the compound 15 is prepared by SN 2 Obtaining a compound 18 through reaction, and reducing the compound 18 by sodium borohydride to obtain a liquid carrier TAG-D;
Figure FDA0004132695200000021
3. the method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 4A-4D is as follows: dichloromethane, a liquid carrier TAG-A, TAG-B, TAG-C or TAG-D, fmoc-Lys (Boc) -OH, N' -diisopropylcarbodiimide and 4-dimethylaminopyridine are sequentially added into a flask, the mixture is stirred for 3 hours, the dichloromethane is concentrated, methanol is added under ice bath, the mixture is stirred for 10 minutes after solid precipitation, and the solid powder compounds 4A-4D are filtered and collected.
4. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 5A-5D is as follows: dichloromethane, the compounds 4A-4D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the flask under ice bath, stirring is continued for 10 minutes after solid precipitation, then filtration is carried out, and solid powder compounds 5A-5D are collected.
5. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 6A-6D is as follows: dichloromethane, fmoc-His (Trt) -OH, N-diisopropylethylamine, N' -diisopropylcarbodiimide and 1-hydroxybenzotriazole are sequentially added into a round bottom flask, after stirring for 10 minutes, the compounds 5A-5D are added into a reaction liquid, stirring is continued for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, and then filtration is carried out, and solid powder compounds 6A-6D are collected.
6. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 7A-7D is as follows: dichloromethane, the compound 6A-6D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a round bottom flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the mixture under ice bath, stirring is continued for 10 minutes after solid precipitation, and the solid powder compound 7A-7D is filtered and collected.
7. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 8A-8D is as follows: dichloromethane, fmoc-Gly-OH, N-diisopropylethylamine, N' -diisopropylcarbodiimide and 1-hydroxybenzotriazole are sequentially added into a round bottom flask, after stirring for 10 minutes, the compounds 7A-7D are added into a reaction liquid, stirring is continued for 3 hours, then dichloromethane is concentrated, methanol is added under ice bath, stirring is continued for 10 minutes after solid precipitation, and then filtering is carried out, and solid powder compounds 8A-8D are collected.
8. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 9A to 9D is as follows: dichloromethane, the compounds 8A-8D and 1, 8-diazabicyclo [5.4.0] undec-7-ene are sequentially added into a round bottom flask, stirred for 30 minutes at room temperature, then dichloromethane is concentrated, acetonitrile is added into the round bottom flask under ice bath, stirring is continued for 10 minutes after solid precipitation, then filtration is carried out, and solid powder compounds 9A-9D are collected.
9. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of the compounds 10A to 10D is as follows: adding dichloromethane, palmitic acid, N-diisopropylethylamine, N' -diisopropylethylamine and 1-hydroxybenzotriazole into a round bottom flask in sequence, stirring for 10 minutes, adding the compounds 9A-9D into a reaction liquid, continuously stirring for 3 hours, concentrating the dichloromethane, adding methanol under ice bath, continuously stirring for 10 minutes after solid precipitation, filtering, collecting solid powder, and drying in a vacuum drying oven at 30 ℃ to obtain white solid compounds 10A-10D.
10. The method for synthesizing palmitoyl tripeptide-1 based on the liquid phase carrier assistance according to claim 1, wherein the synthesis method of palmitoyl tripeptide-1 is as follows: adding dichloromethane and compounds 10A-10D into a round bottom flask in sequence, adding cutting fluid after the reaction fluid is dissolved, continuously stirring for 3 hours, concentrating the reaction fluid, adding isopropyl ether at 0 ℃ to precipitate solids, continuously stirring for 30 minutes, filtering, collecting a filter cake, stirring and dissolving the filter cake with 100 milliliters of water, filtering, preparing and purifying the filtrate, and freeze-drying the collected fluid by a freeze dryer to obtain a white solid final product.
CN202310264615.4A 2023-03-17 2023-03-17 Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance Pending CN116284218A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310264615.4A CN116284218A (en) 2023-03-17 2023-03-17 Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310264615.4A CN116284218A (en) 2023-03-17 2023-03-17 Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance

Publications (1)

Publication Number Publication Date
CN116284218A true CN116284218A (en) 2023-06-23

Family

ID=86784754

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310264615.4A Pending CN116284218A (en) 2023-03-17 2023-03-17 Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance

Country Status (1)

Country Link
CN (1) CN116284218A (en)

Similar Documents

Publication Publication Date Title
CN104004064B (en) A kind of preparation method of buserelin
CN111732649B (en) Preparation of liraglutide by continuous flow solid phase reaction
CN111808165A (en) Synthetic method of GHK copper peptide
CN101357937B (en) Method for synthesizing atosiban acetate from solid phase polypeptide
CN107056894B (en) Method for solid-phase synthesis of ganirelix acetate by fragment method
CN110903352A (en) Preparation method of cetrorelix
CN1990501A (en) Preparing process for synthesizing oxytocin from solid-phase polypeptide
CN113667007B (en) Liquid phase preparation method of cable Ma Lutai side chain
CN111690037A (en) Method for synthesizing GHK acetate
CN116284218A (en) Method for synthesizing palmitoyl tripeptide-1 based on liquid phase carrier assistance
JP2001342200A (en) Method for producing octreotide and its derivative
CN116375792A (en) Short peptide Boc-L-Tyr (tBu) -Aib-OH and preparation method thereof
CN111393508B (en) Preparation method of atosiban
CN105541968B (en) A kind of preparation method and application of pentapeptide
WO1998018815A1 (en) Convergent process for the preparation of a growth hormone secretagogue
CN113667006B (en) Preparation method of cable Ma Lutai dipeptide side chain
CN111285921B (en) BDK auxiliary group and liquid phase total synthesis method of procalcitonin and analog based on BDK auxiliary group
CN112759628A (en) Synthetic method of brennuo peptide
CN113461774A (en) Preparation method of palmitoyl tripeptide-1
CN109293736B (en) Dipeptide for synthesizing relin medicines
CN113801190A (en) Method for preparing oligopeptide-1 hydrochloride
CN113880921B (en) Synthetic method of bremer langdan acetate
CN114685614B (en) Solid-phase synthesis method of atosiban
CN109810169B (en) Liquid phase preparation method of Reltecimod
CN102516362B (en) Method for preparing thymopentin

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination