[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

CN111253287A - Method for synthesizing side chain of Somalutide in liquid phase convergence manner - Google Patents

Method for synthesizing side chain of Somalutide in liquid phase convergence manner Download PDF

Info

Publication number
CN111253287A
CN111253287A CN202010046905.8A CN202010046905A CN111253287A CN 111253287 A CN111253287 A CN 111253287A CN 202010046905 A CN202010046905 A CN 202010046905A CN 111253287 A CN111253287 A CN 111253287A
Authority
CN
China
Prior art keywords
compound
reaction
ethanol
protecting group
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
CN202010046905.8A
Other languages
Chinese (zh)
Inventor
张逢质
杨彬
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.)
Zhejiang University of Technology ZJUT
Original Assignee
Zhejiang University of Technology ZJUT
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 Zhejiang University of Technology ZJUT filed Critical Zhejiang University of Technology ZJUT
Priority to CN202010046905.8A priority Critical patent/CN111253287A/en
Publication of CN111253287A publication Critical patent/CN111253287A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C269/04—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups from amines with formation of carbamate groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C213/00—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton
    • C07C213/02—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton by reactions involving the formation of amino groups from compounds containing hydroxy groups or etherified or esterified hydroxy groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • C07C231/12—Preparation of carboxylic acid amides by reactions not involving the formation of carboxamide groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • C07C231/14—Preparation of carboxylic acid amides by formation of carboxamide groups together with reactions not involving the carboxamide groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C269/06—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups by reactions not involving the formation of carbamate groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/02—Ortho- or ortho- and peri-condensed systems
    • C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
    • C07C2603/06—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members
    • C07C2603/10—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings
    • C07C2603/12—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings only one five-membered ring
    • C07C2603/18—Fluorenes; Hydrogenated fluorenes
    • 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)
  • Peptides Or Proteins (AREA)

Abstract

Liquid phase convergent synthetic soymilkMethod for the Lupeptin side chain 1: the amino group of the raw material 2- (2-aminoethoxy) ethanol is replaced by R1Protecting, nucleophilic substitution reaction with ethyl bromoacetate, ester hydrolyzing in one pot to obtain compound 4, and reacting carboxyl group of compound 4 with R2Protection and removal of R1To obtain a compound 6, carrying out condensation reaction on the compound 6 and fluorenylmethyloxycarbonyl-L-glutamic acid 1-tert-butyl ester to obtain a compound 8, and carrying out condensation reaction on R of the compound 82Removing, performing coupling reaction with compound 6 to obtain compound 10, removing Fmoc of compound 10, performing amidation condensation coupling reaction with 18- (tert-butoxy) -18-oxooctadecanoic acid to obtain compound 13, removing R from compound 132To obtain a product 1; the method has effective and controllable synthesis process, low cost and high yield, and is suitable for large-scale production;

Description

Method for synthesizing side chain of Somalutide in liquid phase convergence manner
Technical Field
The invention belongs to the field of synthesis of polypeptide drugs, and particularly relates to convergent liquid-phase synthesis of a side chain of a polypeptide drug, namely, somaglutide, for treating type II diabetes.
Background
The somaglutide belongs to glucagon-like peptide-1 (GLP-1) receptor agonist, and its molecular formula is C187H291N45O59Molecular weight is 4113.58, CAS number is 910463-68-2. GLP-1 is a hormone inducing insulin secretion, and has beneficial effects on various important organs including pancreas, heart and liver. GLP-1 receptor agonist drugs have the advantages of effectively controlling blood sugar, obviously reducing incidence of hypoglycemic events, obviously reducing weight and reducing risk of cardiovascular events.
Somalutide was approved by the U.S. FDA in 2017 for marketing (0.5mg or 1mg injection) and was used clinically for the treatment of type II diabetes. The us FDA officially approved thaumautide oral insulin to be marketed by us FDA on 9/20/2019 for improving glycemic control in type II diabetic patients in combination with diet and exercise. The once daily oral sommoglutide tablet of norand nordhead is the first oral glucagon-like peptide-1 (GLP-1) to obtain FDA approval.
The structure of the side chain of the somaglutide is as follows:
Figure BDA0002369749250000011
can be seen to consist of 2 molecules of 8-amino-3, 6-dioxaoctanoic acid, one molecule of glutamic acid and one molecule of mono-tert-butyl octadecanedioate. The side chain can enable the somaglutide to be combined with protein more tightly, reduce the rate of clearance by the kidney and prevent rapid metabolic degradation, so that the half life of the drug is prolonged. At present, the somniferin and the side chain thereof are synthesized by coupling and cutting in a solid-phase synthesis mode commonly used in the industry, and the development of liquid-phase synthesis has potential application value in the industry due to poor economical efficiency of solid-phase synthesis atoms.
The invention adopts a liquid phase synthesis method to synthesize the side chain 1 of the Somaloutide.
Disclosure of Invention
The invention aims to synthesize the side chain of the Somaloutide in a low-cost, high-yield and short-period manner by using a convergent liquid phase synthesis manner, and provides a possibility for industrial application.
The final target product of the invention has the following structural formula:
Figure BDA0002369749250000012
the technical scheme of the invention is as follows:
a method for synthesizing a side chain 1 of Somalutide in a liquid phase convergence manner comprises the following steps:
(1) the amino group of the raw material 2- (2-aminoethoxy) ethanol is replaced by R1Protecting, then carrying out nucleophilic substitution reaction with ethyl bromoacetate to prolong a carbon chain, and carrying out ester hydrolysis in one pot to obtain a compound 4;
Figure BDA0002369749250000021
the R is1Is Fmoc, Alloc, Boc, PMB, Cbz, Trt, Tos, Mtt, Mmt, Bom, Sem or MEM, preferably Fmoc, Trt or Boc, more preferably Trt or Boc, R1The reaction solvent used for protection is selected from one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion, preferably ethanol or methanol;
the reaction solvent used for nucleophilic substitution reaction with ethyl bromoacetate is one or a mixture of several solvents in any proportion selected from methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone, and tetrahydrofuran is preferably selected from: methanol 1:1 (v: v);
ester hydrolysis is carried out under the action of inorganic base, wherein the inorganic base is selected from one or a mixture of more of sodium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, calcium hydroxide and barium hydroxide in any proportion, and preferably sodium hydroxide; the mass ratio of the inorganic base to the raw material 2- (2-aminoethoxy) ethanol is 0.5-4: 1, preferably 0.5 to 2: 1 more preferably 2: 1;
(2) the free carboxyl group of the compound 4 is substituted with R2Protection and removal of R1Protecting group to give compound 6;
Figure BDA0002369749250000022
the R is2Bn, Pfp, Me, Allyl, t-Bu, PMB, MEM or TBS, preferably Bn or Me, more preferably Bn;
removal of R1The protecting group is carried out under the action of an acid reagent, wherein the acid reagent is hydrochloric acid, acetic acid or trifluoroacetic acid, and preferably the trifluoroacetic acid; removal of R1The reaction solvent used for the protecting group is selected from one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion, and dichloromethane is preferred; particularly preferably, R is removed1Protecting groups are described in trifluoroacetic acid: volume ratio of dichloromethane 1: 1-2 (preferably 1: 1);
(3) carrying out condensation reaction on the compound 6 and fluorenylmethyloxycarbonyl-L-glutamic acid 1-tert-butyl ester to obtain a compound 8;
Figure BDA0002369749250000023
the solvent for condensation reaction is selected from one or more of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone, preferably 1, 2-dichloroethane or dichloromethane, more preferably dichloromethane; the condensation reaction is carried out under the action of a condensing agent selected from any single or multiple condensing agent of DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT, HATU/HOBT, preferably EDCI/DMAP, EDCI/HOBT or HATU, more preferably EDCI/HOBT, and the ratio of the amounts of EDCI and HOBT is 1: 0.5 to 2, preferably 1: 1; the condensation reaction temperature is 20-70 ℃, preferably 20-40 ℃, and more preferably 30 ℃;
(4) r of compound 82Removing the protecting group, and then carrying out coupling reaction with a compound 6 to obtain a compound 10;
Figure BDA0002369749250000024
R2the method for removing the protecting group by hydrogenation uses palladium carbon, wherein the mass of palladium in the palladium carbon accounts for 5 percent of the total mass of the palladium carbon, and the using amount of the palladium accounts for 1 to 20 percent, preferably 5 to 15 percent and more preferably 5 percent of the mass of the compound 8; r2The reaction solvent used for removing the protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion, preferably methanol or acetonitrile, more preferably methanol;
the solvent for coupling reaction is selected from one or more of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone, preferably 1, 2-dichloroethane or dichloromethane, more preferably dichloromethane; the coupling reaction is carried out with the aid of a condensing agent selected from any single or complex condensing agent of DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT, HATU/HOBT, preferably EDCI/DMAP, EDCI/HOBT or HATU, more preferably EDCI/HOBT, and the mass ratio of EDCI and HOBT is 1: 0.5 to 2, preferably 1: 1; the temperature of the coupling reaction is 20-70 ℃, preferably 20-40 ℃, and more preferably 30 ℃;
(5) after removing Fmoc protecting group of compound 10, carrying out amidation condensation coupling reaction with 18- (tert-butoxy) -18 oxooctadecanoic acid to obtain compound 13;
Figure BDA0002369749250000031
the removal of the Fmoc protecting group is performed under the action of an organic base selected from diethylamine, N-diisopropylethylamine, triethylamine, piperidine, imidazole, pyridine or DBU, preferably triethylamine, DBU or diethylamine, more preferably diethylamine; the reaction solvent for removing the Fmoc protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion, preferably acetonitrile or methanol, more preferably acetonitrile;
the solvent for amidation condensation coupling reaction is selected from one or more of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone, and is a mixed solvent in any proportion, 1, 2-dichloroethane or dichloromethane is preferred, and dichloromethane is more preferred; the amidated condensation coupling reaction is carried out with the aid of a condensing agent selected from any single or complex condensing agent of DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT, HATU/HOBT, preferably EDCI/DMAP, EDCI/HOBT or HATU, more preferably EDCI/HOBT, and the ratio of the amounts of EDCI and HOBT is 1: 0.5 to 2, preferably 1: 1; the temperature of the amidation condensation coupling reaction is 20-70 ℃, preferably 20-40 ℃, and more preferably 30 ℃;
(6) removal of R from Compound 132Protecting groups to obtain a final target product 1;
Figure BDA0002369749250000032
R2the removal of the protecting group is carried out under the action of an inorganic baseSelected from potassium hydroxide, barium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide or zinc hydroxide, preferably potassium hydroxide or lithium hydroxide, more preferably lithium hydroxide, preferably the inorganic base is dosed in the form of an aqueous solution, e.g. 0.5-2M (preferably 1M) aqueous lithium hydroxide solution; removal of R2The reaction solvent used for the protecting group is selected from one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion, preferably ethanol or methanol, and more preferably methanol.
The final synthesized product of the invention can directly perform condensation reaction with free amino groups on the somaglutide Lys residues so as to be connected to the somaglutide main chain.
The invention has the advantages that: the convergent synthesis method is used for reducing the reaction cost and shortening the reaction time. The synthesis process is effective and controllable, has low cost and high yield, and is suitable for large-scale production.
The abbreviations used in the present invention have the following meanings:
his: histidine
Aib: 2-methylalanine
Glu: glutamic acid
Gly: glycine
Thr: threonine
Phe: phenylalanine
Thr: threonine
Ser: serine
Asp: aspartic acid
Val: valine
Leu: leucine
Gln: glutamine
Ala: alanine
Lys: lysine
Fmoc: fmoc group
And (3) Alloc: allyloxycarbonyl radical
Boc: tert-butyloxycarbonyl radical
PMB: p-methoxybenzyl
Trt: trityl radical
Tos: p-toluenesulfonyl group
Mtt: 4-methyl-trityl radical
Mmt: 4-Methoxytrityl group
And (5) Sem: trimethylsiloxyethylmethyl group
MEM: 2-methoxyethoxymethyl group
EDCI: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride
HOBT: 1-hydroxybenzotriazoles
HATU: o- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea
DIC: n, N' -diisopropylcarbodiimide
DCC: dicyclohexylcarbodiimide
HBTU: benzotriazole-1-tetramethylhexafluorophosphate
DMAP: 4-dimethylaminopyridine
PyBOP: 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphates
BOP: benzotriazole-1-tris (trimethylamino) -trifluorophosphate
TBTU: o- (1H-benzotriazol-1-yl) -N, N, N ', N' -tetramethylisourea boron tetrafluoride
Drawings
FIG. 1 is a synthesis scheme of example 1 of the present invention.
FIG. 2 is a MS plot of Compound 1.
FIG. 3 is an IR spectrum of Compound 1
Detailed Description
The present invention is further illustrated by the following specific examples, but the scope of the invention is not limited thereto.
With R1＝Boc，R2Liquid phase synthesis, with Bn as specific example 1, was performed according to the scheme for the synthesis of the side chain of somaglutide of figure 1
Example 1-1:
one-pot preparation
Figure BDA0002369749250000041
Dissolving 2- (2-aminoethoxy) ethanol 2 serving as a starting material (300mmol, 31.5g) in 500mL of ethanol solution, slowly dropwise adding di-tert-butyl dicarbonate (300mmol, 69mL) under the ice bath condition, removing the ice bath, reacting at room temperature for 2 hours, and distilling under reduced pressure to remove the ethanol solvent; dissolving tetrahydrofuran again, adding sodium hydride (1.2-1.5eq) at 0-10 ℃, keeping the ice bath and stirring for half an hour, slowly dripping bromoethyl acetate 3(1.2-1.5eq) into the reaction solution, removing the ice bath after finishing dripping, and stirring at room temperature overnight. MeOH was added to the reaction, and MeOH: THF 1:1, the reaction solution was yellow, clear and transparent. Solid NaOH (1-2.2eq) is weighed and added into the reaction solution, and the reaction solution is heated and refluxed for 2 hours. And after the reaction is finished, distilling under reduced pressure to remove the solvent, re-dissolving the residue with water, extracting twice with ethyl acetate, collecting a water layer, adjusting the pH value to 1-3, extracting twice with ethyl acetate, collecting an organic layer, and distilling under reduced pressure to obtain a yellow oily liquid 4. The yield thereof was found to be 82%.
Examples 1 to 2:
Figure BDA0002369749250000051
and (4) preparing.
(1) Under ice-bath conditions, benzyl chloroformate 5(100mmol, 12.9mL) was slowly added dropwise to a solution of compound 4(1-1.2eq) and triethylamine (1-1.2eq) in dichloromethane (100mL), and after stirring for 5min, DMAP (0.1-0.3eq) was added to the reaction mixture, followed by overnight reaction at room temperature and monitoring by TLC (CH)2Cl2: EA 10: 1). After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was redissolved with ethyl acetate, extracted with 10% citric acid (75 mL. times.2) and saturated sodium bicarbonate solution (75 mL. times.2), washed with saturated brine (100mL), and the organic layer was collected and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain a colorless oily liquid. The yield thereof was found to be 63%.
(2) The above colorless oily liquid was dissolved in a dichloromethane solution using trifluoroacetic acid: dichloromethane (v: v) ═ 1:1 was deprotected, and trifluoroacetic acid was distilled off under reduced pressure to give yellow oily liquid 6 for use.
Examples 1 to 3:
Figure BDA0002369749250000052
and (4) preparing.
Fmoc-L-glutamic acid 1-tert-butyl ester (1g, 2.35mmol) was weighed and dissolved in 25mL of dichloromethane, DIPEA (3eq, 1.2mL), the condensing agents EDCI (1.2eq, 541mg) and HOBT (1.2eq, 380mg) were added in this order, and after stirring for 30min, Compound 6(1.2eq) was added, stirring was carried out at 30 ℃, and the reaction was monitored by TLC. After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was redissolved with ethyl acetate, extracted with 10% aqueous citric acid solution and saturated sodium bicarbonate solution, washed with saturated brine (75mL), and the organic layer was collected and dried over anhydrous sodium sulfate. The solvent was removed by distillation under the reduced pressure, and the residue was purified by column chromatography to give yellow oily substance 8. The yield thereof was found to be 92%.
Examples 1 to 4:
Figure BDA0002369749250000053
and (4) preparing.
(1) Debenzylation; dissolving 2.64g (4mmol) of compound 8 in 15mL of methanol solution, placing the mixture in a three-neck flask, replacing air with nitrogen, adding a catalytic amount of 5% palladium-carbon (the mass fraction of palladium is 5%) 424mg (0.2mmol), inflating a balloon with hydrogen, replacing nitrogen in the flask, reacting at room temperature for 2 hours, filtering off the palladium-carbon by suction, and performing spin-drying to obtain compound 9 for later use with the yield of 90%.
(2) 2.05g of Compound 9 was dissolved in 25mL of methylene chloride, DIPEA (3eq, 1.8mL), the condensing agents EDCI (1.2eq, 830mg) and HOBT (1.2eq, 583mg) were added in this order, and after stirring for 30min, Compound 6(1.2eq) was added, stirring was carried out at 30 ℃ and the reaction was monitored by TLC. After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was redissolved with ethyl acetate, extracted with 10% citric acid (50 mL. times.2) and saturated sodium bicarbonate solution (50 mL. times.2), washed with saturated brine (75mL), and the organic layer was collected and dried over anhydrous sodium sulfate. The solvent was removed by distillation under the reduced pressure and purified by column Chromatography (CH)2Cl2: MeOH ═ 20:1) gave a yellow oil, 10. The yield thereof was found to be 85%.
Examples 1 to 5:
Figure BDA0002369749250000061
and (4) preparing.
The above starting material 10(1.7mmol, 1.36g) was dissolved in 30-40mL of anhydrous acetonitrile, diethylamine (3-4mL) was added dropwise, stirred at room temperature for 3h, and the reaction monitored by TLC. Purification by column Chromatography (CH)2Cl2: MeOH 10:1) gave 11 as a yellow oil. The yield thereof was found to be 66%.
Examples 1 to 6:
Figure BDA0002369749250000062
and (4) preparing.
A25 mL round bottom flask was taken and 18- (tert-butoxy) -18 oxooctadecanoic acid (0.83mmol, 308mg) was dissolved in 5mL of dichloromethane, followed by the sequential addition of DIPEA (3eq, 0.41mL), the condensing agent EDCI/HOBT (1.2eq), followed by stirring for 30min and the addition of intermediate 11(0.1mmol, 583mg) above, and the reaction was monitored by TLC. After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was redissolved with ethyl acetate, extracted with 10% citric acid (10 mL. times.2) and saturated sodium bicarbonate solution (10 mL. times.2), washed with saturated brine (20mL), and the organic layer was collected and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain an oily liquid 13 in a yield of 82%.
Examples 1 to 7:
Figure BDA0002369749250000063
and (4) preparing.
Taking a 25mL round-bottom flask, dissolving the intermediate 13 in 2mL MeOH, dropwise adding 1mol/L LiOH aqueous solution into the reaction solution, stirring at room temperature for 2h, removing the solvent by reduced pressure distillation, and purifying by column Chromatography (CH)2Cl2: MeOH ═ 5:1) to give final product 1. The yield thereof was found to be 90%.
With R1＝Trt，R2As specific example 2
Example 2-1:
Figure BDA0002369749250000064
and (4) preparing.
Dissolving 2- (2-aminoethoxy) ethanol 2 as a starting material (100mmol, 10.5g) in 150mL of dichloromethane solution, slowly dropwise adding triethylamine (100mmol, 14mL) under the condition of ice bath, keeping the temperature, stirring for half an hour, slowly dropwise adding Trt-Cl (1.2eq) into the reaction solution, removing the ice bath after dropwise adding is finished, and stirring for 1h at room temperature. After the reaction, the solvent was distilled off under reduced pressure. THF is redissolved, potassium carbonate (2eq) is added at 0 to-10 ℃, the temperature is kept and stirring is carried out for half an hour, benzyl bromoacetate (1.2 to 1.5eq) is slowly dropped into the reaction solution, the ice bath is removed after the dropping is finished, and stirring is carried out at 50 ℃ overnight. After the reaction, the solvent was distilled off under reduced pressure, extracted twice with ethyl acetate, washed with saturated brine, and the organic layer was collected and dried over anhydrous sodium sulfate. And carrying out reduced pressure distillation to obtain a target product. The yield thereof was found to be 90%.
Example 2-2:
Figure BDA0002369749250000071
and (4) preparing.
The product was dissolved in dichloromethane solution using trifluoroacetic acid: dichloromethane (v: v) ═ 1:1 was deprotected, and trifluoroacetic acid was distilled off under reduced pressure to give yellow oily liquid 6 for use.
The subsequent procedure for the preparation of the final product 1 was the same as in example 1.
This particular embodiment is provided to complement and illustrate the feasibility of the present invention, and many other variations and modifications may be made without departing from the spirit and scope of the present invention. Therefore, the scope of the appended claims should also encompass variations and modifications on the basis of the present invention.
The invention provides a convergent liquid-phase synthesis mode to complete the high-efficiency synthesis of the Somaloutide side chain 1, thereby reducing the reaction cost and providing possibility for industrial production.

Claims (7)

1. A method for synthesizing a side chain 1 of Somalutide in a liquid phase convergence manner is characterized by comprising the following steps:
(1) the amino group of the raw material 2- (2-aminoethoxy) ethanol is replaced by R1Protection, nucleophilic substitution reaction with ethyl bromoacetate to elongate carbon chain, and reactionHydrolyzing the ester in one pot to obtain a compound 4;
Figure FDA0002369749240000011
(2) the free carboxyl group of the compound 4 is substituted with R2Protection and removal of R1Protecting group to give compound 6;
Figure FDA0002369749240000012
(3) carrying out condensation reaction on the compound 6 and fluorenylmethyloxycarbonyl-L-glutamic acid 1-tert-butyl ester to obtain a compound 8;
Figure FDA0002369749240000013
(4) r of compound 82Removing the protecting group, and then carrying out coupling reaction with a compound 6 to obtain a compound 10;
Figure FDA0002369749240000014
(5) after removing Fmoc protecting group of compound 10, carrying out amidation condensation coupling reaction with 18- (tert-butoxy) -18 oxooctadecanoic acid to obtain compound 13;
Figure FDA0002369749240000015
(6) removal of R from Compound 132Protecting groups to obtain a final target product 1;
Figure FDA0002369749240000016
2. the liquid phase convergent method for synthesizing the side chain 1 of Somalutide according to claim 1, wherein in step (1), R is1Is Fmoc, Alloc, Boc, PMB,Cbz, Trt, Tos, Mtt, Mmt, Bom, Sem or MEM, R1The reaction solvent used for protection is selected from one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion;
the reaction solvent used for nucleophilic substitution reaction with ethyl bromoacetate is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion;
ester hydrolysis is carried out under the action of inorganic base, wherein the inorganic base is one or a mixture of more of sodium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, calcium hydroxide and barium hydroxide in any proportion; the mass ratio of the inorganic base to the raw material 2- (2-aminoethoxy) ethanol is 0.5-4: 1.
3. the liquid phase convergent method for synthesizing the side chain 1 of Somalutide according to claim 1, wherein in step (2), R is2Bn, Pfp, Me, Allyl, t-Bu, PMB, MEM or TBS;
removal of R1The protecting group is carried out under the action of an acid reagent, wherein the acid reagent is hydrochloric acid, acetic acid or trifluoroacetic acid; removal of R1The reaction solvent used for the protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion.
4. The liquid phase convergent method for synthesizing the Somalutide side chain 1 according to claim 1, wherein in the step (3), the solvent for the condensation reaction is selected from one or more of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone; the condensation reaction is carried out under the action of a condensing agent, and the condensing agent is any single condensing agent or composite condensing agent selected from DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT and HATU/HOBT; the temperature of the condensation reaction is 20-70 ℃.
5. The liquid phase convergent method for synthesizing the side chain 1 of Somalutide according to claim 1, wherein in step (4), R is2The method for removing the protecting group by using palladium-carbon hydrogenation comprises the following steps of (1) removing the protecting group by using palladium-carbon, wherein the mass of palladium in the palladium-carbon accounts for 5% of the total mass of the palladium-carbon, and the using amount of palladium accounts for 1% -20% of the amount of substances of a compound 8; r2The reaction solvent used for removing the protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion;
the solvent for coupling reaction is selected from one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion; the coupling reaction is carried out under the action of a condensing agent, wherein the condensing agent is any single condensing agent or composite condensing agent selected from DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT and HATU/HOBT; the temperature of the coupling reaction is 20-70 ℃.
6. The liquid phase convergent method for synthesizing the somaglutide side chain 1 according to claim 1, wherein in step (5), the Fmoc-protecting group removal is performed under the action of an organic base selected from diethylamine, N-diisopropylethylamine, triethylamine, piperidine, imidazole, pyridine or DBU; the reaction solvent for removing the Fmoc protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion;
the solvent for amidation condensation coupling reaction is one or more of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion; the amidation condensation coupling reaction is carried out under the action of a condensing agent, wherein the condensing agent is any single condensing agent or composite condensing agent selected from DIC, DCC, HBTU, PyBOP, BOP, HATU, TBTU, DIC/HOBT, DCC/DMAP, EDCI/HOBT and HATU/HOBT; the temperature of the amidation condensation coupling reaction is 20-70 ℃.
7. The liquid phase convergent method for synthesizing the side chain 1 of Somalutide according to claim 1, wherein in step (6), R is2The removal of the protecting group is carried out under the action of an inorganic base selected from potassium hydroxide, barium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide or zinc hydroxide; removal of R2The reaction solvent used for the protecting group is one or a mixture of several of methanol, ethanol, ethyl acetate, dichloromethane, 1, 2-dichloroethane, DMF, DMSO, tetrahydrofuran, acetonitrile and N-methylpyrrolidone in any proportion.
CN202010046905.8A 2020-01-16 2020-01-16 Method for synthesizing side chain of Somalutide in liquid phase convergence manner Pending CN111253287A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010046905.8A CN111253287A (en) 2020-01-16 2020-01-16 Method for synthesizing side chain of Somalutide in liquid phase convergence manner

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010046905.8A CN111253287A (en) 2020-01-16 2020-01-16 Method for synthesizing side chain of Somalutide in liquid phase convergence manner

Publications (1)

Publication Number Publication Date
CN111253287A true CN111253287A (en) 2020-06-09

Family

ID=70944029

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010046905.8A Pending CN111253287A (en) 2020-01-16 2020-01-16 Method for synthesizing side chain of Somalutide in liquid phase convergence manner

Country Status (1)

Country Link
CN (1) CN111253287A (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113461591A (en) * 2021-08-12 2021-10-01 浙江泽瑞生物医药有限公司 Preparation method of precursor raw material Pht-AEEA-AEEA for side chain of Somalutide
CN113896657A (en) * 2021-11-16 2022-01-07 山东盛安贝新能源有限公司南京分公司 Synthesis method and purification method of somaglutide protected amino acid
CN114213283A (en) * 2022-01-04 2022-03-22 攀枝花学院 Method for preparing [2- [1- (Fmoc-amino) ethoxy ] acetic acid by one-pot method
CN114805543A (en) * 2022-04-29 2022-07-29 四川普康药业有限公司 Synthesis method of somaglutide side chain
CN115197312A (en) * 2022-08-09 2022-10-18 奥锐特药业(天津)有限公司 Preparation method and intermediate of Somalutide

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105154498A (en) * 2007-12-28 2015-12-16 诺沃—诺迪斯克有限公司 Semi-recombinant preparation of GLP-1 analogues
CN105683156A (en) * 2013-08-29 2016-06-15 佩特雷化学和生物制药学实验室有限公司 Amino diacids containing peptide modifiers
CN105745222A (en) * 2013-10-17 2016-07-06 西兰制药公司 Acylated glucagon analogues
CN110041219A (en) * 2019-05-09 2019-07-23 南京工业大学 Liquid phase synthesis method of side chain of Somaloutide

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105154498A (en) * 2007-12-28 2015-12-16 诺沃—诺迪斯克有限公司 Semi-recombinant preparation of GLP-1 analogues
CN105683156A (en) * 2013-08-29 2016-06-15 佩特雷化学和生物制药学实验室有限公司 Amino diacids containing peptide modifiers
CN105745222A (en) * 2013-10-17 2016-07-06 西兰制药公司 Acylated glucagon analogues
CN110041219A (en) * 2019-05-09 2019-07-23 南京工业大学 Liquid phase synthesis method of side chain of Somaloutide

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
东圆珍等: "索玛鲁肽的制备", 《中国医药工业杂志》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113461591A (en) * 2021-08-12 2021-10-01 浙江泽瑞生物医药有限公司 Preparation method of precursor raw material Pht-AEEA-AEEA for side chain of Somalutide
CN113896657A (en) * 2021-11-16 2022-01-07 山东盛安贝新能源有限公司南京分公司 Synthesis method and purification method of somaglutide protected amino acid
CN114213283A (en) * 2022-01-04 2022-03-22 攀枝花学院 Method for preparing [2- [1- (Fmoc-amino) ethoxy ] acetic acid by one-pot method
CN114805543A (en) * 2022-04-29 2022-07-29 四川普康药业有限公司 Synthesis method of somaglutide side chain
CN115197312A (en) * 2022-08-09 2022-10-18 奥锐特药业(天津)有限公司 Preparation method and intermediate of Somalutide
WO2024032081A1 (en) * 2022-08-09 2024-02-15 扬州奥锐特药业有限公司 Preparation method for semaglutide, and intermediate

Similar Documents

Publication Publication Date Title
CN111253287A (en) Method for synthesizing side chain of Somalutide in liquid phase convergence manner
CN110041219B (en) Liquid phase synthesis method of side chain of Somaloutide
JP6500105B2 (en) Peptide-bound stable ascorbic acid derivative, method for producing the same, and cosmetic composition containing the same
CN111269137A (en) Method for preparing side chain of Somalutide by liquid phase method
CN112625087A (en) Dipeptide fragment derivative for synthesizing semaglutide and preparation method thereof
CN111378028A (en) Synthesis of acylated GLP-1 compounds and modified groups thereof
AU2019433421B2 (en) Preparation method for and intermediate of drug-linker for antibody drug conjugate MC-MMAF
CN109232743B (en) Long-acting hypoglycemic weight-loss peptide, preparation method thereof and application thereof as medicine
CN114685646B (en) Preparation method and application of polypeptide side chain analogue
TW202235429A (en) Method for preparing peptide compounds containing n-substited amino acid residue
CZ288529B6 (en) Cyclic amino acids, process for preparing D- and L-enantiomers thereof and intermediates for their preparation
CN114213503A (en) Synthesis method of snake venom-like tripeptide
CN107056926B (en) Exendin-4 analogues with ether bonds and application thereof
CN109879936B (en) Method for synthesizing palmitoyl hexapeptide by using micro-channel modular reaction device
CN108530518A (en) 10 analog of aplysiatoxin and its preparation method and application
CN105418737B (en) Solid-phase synthesis method and application of bregma cyclopeptide A
CN111454180A (en) Somalutide side chain intermediate and preparation method thereof
WO1991001976A1 (en) Thioacylating reagents and intermediates, thiopeptides, and methods for preparing and using same
CN111285780A (en) Convergent liquid phase synthesis method of side chain of Somaloutide
CN107298708B (en) Glucagon-like peptide-1 (GLP-1) analogue with ether bond and application thereof
CN116496345A (en) Polypeptide ceramide and synthesis method and application thereof
CN110183532B (en) Process method for synthesizing bivalirudin protected pentapeptide fragments by mass high-efficiency liquid phase method
CN111285921B (en) BDK auxiliary group and liquid phase total synthesis method of procalcitonin and analog based on BDK auxiliary group
FR2756566A1 (en) BRADYKININ B2 RECEPTOR B2 AGONIST PEPTIDES, PREPARATION METHOD AND THERAPEUTIC USE
CN112679408A (en) Synthesis method of side chain derivative of somasu peptide fatty acid

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
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20200609