WO2008009672A1 - Process for the manufacture of peptides - Google Patents

Process for the manufacture of peptides Download PDF

Info

Publication number
WO2008009672A1
WO2008009672A1 PCT/EP2007/057362 EP2007057362W WO2008009672A1 WO 2008009672 A1 WO2008009672 A1 WO 2008009672A1 EP 2007057362 W EP2007057362 W EP 2007057362W WO 2008009672 A1 WO2008009672 A1 WO 2008009672A1
Authority
WO
WIPO (PCT)
Prior art keywords
process according
group
merrifield
amino
peptide
Prior art date
Application number
PCT/EP2007/057362
Other languages
French (fr)
Inventor
Satish Joshi
Shima Joshi
Bernhard Freimann
Manuel Ramos
Roland Callens
Original Assignee
Solvay (Societe Anonyme)
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 Solvay (Societe Anonyme) filed Critical Solvay (Societe Anonyme)
Priority to CN200780034795.6A priority Critical patent/CN101516903B/en
Priority to EP07787628A priority patent/EP2049561A1/en
Priority to US12/374,344 priority patent/US8415454B2/en
Priority to JP2009521219A priority patent/JP5220012B2/en
Publication of WO2008009672A1 publication Critical patent/WO2008009672A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/50—Cyclic peptides containing at least one abnormal peptide link
    • C07K7/54—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring
    • C07K7/56—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring the cyclisation not occurring through 2,4-diamino-butanoic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers

Definitions

  • the present invention concerns a process for the manufacture of peptides and intermediates useful therein.
  • Peptides are potentially biologically active and quite a lot of useful peptide-based drugs are known today.
  • a particular example of a biologically active peptide is Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-OH
  • PT- 141 which is a candidate drug useful for treatment of erectile dysfunction (Drugs Fut 2004, 29(6), p.645).
  • Different synthetic approaches have been developed for this molecule. For example, Flora et al. Bioorg. Med Chem. Lett. 2005 (1065-1068) describes a solid phase peptide synthesis strategy for the molecule. However, this process requires use of costly Wang type resins and extremely costly protection and deprotection technology.
  • the invention concerns in consequence a process for manufacturing a cyclic peptide which comprises
  • the process according to the invention allows for industrial scale manufacture of cyclic peptides or peptide derivatives with reagents having low cost.
  • the cyclic peptide obtained in the process according to the invention has high purity and can be purified more easily, use of HF commonly used for removing peptide from Merrifield-type resins is avoided according to the process according to the invention.
  • This is particularly advantageous, as the use of HF is an obstacle to industrial scale peptide synthesis with Merrifield-type resins.
  • Merrifield-type resin intends to denote in particular a chloromethylated styrene-divinylbenzene copolymer, which can optionally be substituted by substituents.
  • An unsubstituted chloromethylated styrene-divinylbenzene copolymer is more particularly preferred.
  • the cyclic structure of the cyclic peptide is generally formed by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 14, 15, 16, 17, 18, 19 or 20 amino acids.
  • the cyclic structure of the cyclic peptide is preferably formed by 5, 6, 7, 8, 9, 10, 11 or 12 amino acids and more preferably by 6 or 7 amino acids.
  • the cyclic peptide has preferably an amide bond formed through intramolecular reaction of a carboxylic group with an amino group.
  • the transesterification is advantageously carried out with an alcohol or a mercaptane on particular an alkyl alcohol or an alkyl mercaptane, in particular a C1-C3 alcohol, preferably selected from methanol and ethanol. Methanol is more particularly preferred. Methanethiol or ethanethiol is preferred as mercaptane, in particular in the form of their alkali metal salts.
  • the transesterification is carried out with an alcohol, the cyclized supported peptide is preferably suspended in the alcohol or in a mixture of the alcohol with a supplementary suitable solvent.
  • the cyclized supported peptide is preferably suspended in a solution of a salt of the mercaptane in a suitable solvent.
  • a transesterification catalyst may suitably be used which can be selected, for example, from bases, in particular nucleophilic bases such as for example amines, in particular tertiary amines or sterically hindered alkoxides such as tert. alkoxides e.g potassium tert.-butoxide, anion-exchange resins in particular quaternary anion-exchange resins and alkalimetal salts of nuclephilic anions selected in particular from cyanides and fluorides such as potassium cyanide or potassium fluoride.
  • Tertiary amines are preferred in particular diisopropylethyl amine and most preferably triethyl amine.
  • the concentration in the mixture catalyst/solvent is generally from 0.5 to 3 mole/liter, preferably from 1 to 2.5 mole/liter and most preferably about 2 mole/liter.
  • the transesterification is carried out with an aminoalcohol, in particular an aminoalcohol having a tertiary amine function and a primary alcohol function such as in particular 2-dimethylaminoethanol.
  • use of supplemental catalyst is not required as the amino function catalyses the reaction.
  • the use of toxic catalysts such as thallium derivatives as in Savoie et al. Can J. Chem. 1974 p. 2832-9 can be avoided.
  • the transesterification is generally carried out at a temperature of from 0 0 C to 100 0 C and preferably from 10 0 C to 60 0 C. A temperature of about 25°C is particularly preferred.
  • the transesterification is generally carried out at atmospheric pressure.
  • the free cyclic peptide acid can suitably be obtained from a cyclic peptide ester obtained in step (b) by reaction with a base selected for example from alkaline, quaternary ammonium or earth- alkaline metal hydroxides or carbonates.
  • a base selected for example from alkaline, quaternary ammonium or earth- alkaline metal hydroxides or carbonates.
  • Particular examples include tetrabutylammonium hydroxide, potassium carbonate and lithium hydroxide. Lithium hydroxide is preferred.
  • cleavage is generally carried out at a pH of from 8 to 10.
  • the optional cleavage is generally carried out at a temperature of from 0 0 C to 100 0 C and preferably from 10 0 C to 60 0 C. A temperature of about 25°C is particularly preferred.
  • the invention concerns a process for manufacturing a cyclic peptide having an amide bond formed through intramolecular reaction of a carboxylic group with an amino group which comprises
  • the linear peptide is generally cyclized in the presence of an amide bond forming reagent.
  • the amide bond forming reagent is suitably - A -
  • amide bond forming agents are selected from mixed anhydrides and other acylating agents such as activated esters or acid halogenides for example isobutyl-chloroformate or pivaloyl chloride. They may be carbodiimides
  • a preferred amide bond forming reagent is dicylohexylcarbodiimide. It has been found that this cheap reagent gives good results in the cyclization.
  • the process according to the invention further comprises providing a precursor linear peptide having a protected amino group and a protected carboxylic group.
  • the amino group is protected by a first protective group.
  • the first protective group is selected for example from protective groups which can be removed under conditions leaving the bond of the linear peptide to the Merrif ⁇ eld resin substantially intact.
  • Preferred examples of such protective groups include amino protective groups which can be removed by acidolysis selected for example from t.butoxycarbonyl, para-methoxybenzyloxycarbonyl, benzhydryloxycarbonyl, and, preferably, benzyloxycarbonyl.
  • protective groups include amino protective groups which can be removed by reduction with metals or metal compounds such as for example trichloroethoxycarbonyl.
  • the carboxylic group is protected by a second protective group.
  • the second protective group is selected for example from protective groups which can be removed under conditions leaving the bond of the linear peptide to the Merrif ⁇ eld resin substantially intact.
  • Preferred examples of such protective groups include carboxyl protective groups which can be removed by acidolysis selected for example from para-methoxybenzyl, benzhydryl, and, preferably, t.butyl.
  • protective groups include protective groups which can be removed by reduction with metals or metal compounds such as for example trichloroethyl.
  • first and second protective groups are preferably selected to be removable in a single reaction step. Accordingly, the protective groups of the precursor peptide can be removed in a single reaction to provide the linear peptide.
  • suitable combinations of first and second protective groups which can be removed by acidolysis a single reaction step include benzhydryloxycarbonyl/benzhydryl; para-methoxybenzyloxycarbonyl/ para-methoxybenzyl and, preferably, t.butoxycarbonyl/ t.butyl.
  • suitable combinations of first and second protective groups which can be removed by reduction with metal or metal compounds in a single reaction step include trichloroethoxycarbonyl/ trichloroethyl
  • suitable reagents are selected for example from perfluoroalkyl carboxylic acids and solutions of HCl in organic solvents preferably polar organic solvents such as dioxane or ethyl acetate. Trifluoroacetic acid is preferred as reagent for acidolysis.
  • a suitable reagent is, for example, zinc metal.
  • the process according to the invention is particularly suitable when the linear peptide comprises an amino acid having a side chain containing an amino group.
  • amino acid having a side chain containing an amino group include lysine, ornithine, diaminobutyric acid and diaminopropionic acid.
  • a preferred example of amino acid having a side chain containing an amino group is lysine.
  • the linear peptide is bonded to the Merrifield resin through the amino acid having a side chain containing an amino group.
  • the linear peptide can suitably comprise an amino acid having a side chain containing a carboxylic group.
  • amino acids having a side chain containing a carboxylic group include glutamic acid and aspartic acid.
  • a preferred example is aspartic acid.
  • the linear peptide comprises both an amino acid having a side chain containing a carboxylic group and an amino acid having a side chain containing an amino group.
  • the side chain carboxylic group or side chain amino group can be suitably protected by the respective protective groups described above.
  • the invention concerns in particular a process for the manufacture of Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-OH
  • the supported cyclic peptide according to the invention is preferably as described herein before.
  • the invention also concerns P-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin I I wherein P is an amino protecting group, in particular an acyl group and preferably an acetyl group (Ac).
  • the invention also concerns
  • P-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin wherein P is as defined here before.
  • Fmoc SPPS strategy used for synthesis at a concentration of 10 ml/gm.
  • Amino acid derivatives used for synthesis The following Amino acid derivatives used for synthesis :
  • the Peptide was cleaved from the resin utilizing the following procedure : Peptide resin stirred for 2 days at ambient temperature in 15 % TEA in Methanol. After cleavage from the resin 5 % DI water and 5eq. LiOH added and continued to stir for 2 hours. Acetic acid was added to acidify reaction mixture. Resin filtered off and washed with TFA A Buffer containing 30 % Acetonitrile. Filtrate evaporated or diluted with TFA A Buffer until organic solvent component was below 20 %. Solution filtered and purified. Purification :

Landscapes

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

Abstract

A process for manufacturing a cyclic peptide which comprises providing a cyclic peptide bonded to a Merrifield-type resin and cleaving the cyclic peptide from the Merrifield type resin by transesterification.

Description

Process for the manufacture of peptides
The present application claims benefit of US provisional 60/832,649, the contents of which is incorporated by reference into the present application.
The present invention concerns a process for the manufacture of peptides and intermediates useful therein.
Peptides are potentially biologically active and quite a lot of useful peptide-based drugs are known today. A particular example of a biologically active peptide is Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-OH
(PT- 141) which is a candidate drug useful for treatment of erectile dysfunction (Drugs Fut 2004, 29(6), p.645). Different synthetic approaches have been developed for this molecule. For example, Flora et al. Bioorg. Med Chem. Lett. 2005 (1065-1068) describes a solid phase peptide synthesis strategy for the molecule. However, this process requires use of costly Wang type resins and extremely costly protection and deprotection technology.
It is an object of the present invention to provide an industrializable process for manufacture of cyclic peptides having improved yield of cyclic product.
The invention concerns in consequence a process for manufacturing a cyclic peptide which comprises
(a) providing a cyclic peptide bonded to a Merrifield-type resin and
(b) cleaving the cyclic peptide from the Merrifield type resin by transesterification.
It has been found, surprisingly, that the process according to the invention allows for industrial scale manufacture of cyclic peptides or peptide derivatives with reagents having low cost. The cyclic peptide obtained in the process according to the invention has high purity and can be purified more easily, use of HF commonly used for removing peptide from Merrifield-type resins is avoided according to the process according to the invention. This is particularly advantageous, as the use of HF is an obstacle to industrial scale peptide synthesis with Merrifield-type resins. For the purposes of the present invention "Merrifield-type resin" intends to denote in particular a chloromethylated styrene-divinylbenzene copolymer, which can optionally be substituted by substituents. An unsubstituted chloromethylated styrene-divinylbenzene copolymer is more particularly preferred.
In the process according to the invention, the cyclic structure of the cyclic peptide is generally formed by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 14, 15, 16, 17, 18, 19 or 20 amino acids. The cyclic structure of the cyclic peptide is preferably formed by 5, 6, 7, 8, 9, 10, 11 or 12 amino acids and more preferably by 6 or 7 amino acids.
In the process according to the invention, the cyclic peptide has preferably an amide bond formed through intramolecular reaction of a carboxylic group with an amino group.
In the process according to the invention, the transesterification is advantageously carried out with an alcohol or a mercaptane on particular an alkyl alcohol or an alkyl mercaptane, in particular a C1-C3 alcohol, preferably selected from methanol and ethanol. Methanol is more particularly preferred. Methanethiol or ethanethiol is preferred as mercaptane, in particular in the form of their alkali metal salts. When the transesterification is carried out with an alcohol, the cyclized supported peptide is preferably suspended in the alcohol or in a mixture of the alcohol with a supplementary suitable solvent.
When the transesterification is carried out with a mercaptane, the cyclized supported peptide is preferably suspended in a solution of a salt of the mercaptane in a suitable solvent.
In the process according to the invention, a transesterification catalyst may suitably be used which can be selected, for example, from bases, in particular nucleophilic bases such as for example amines, in particular tertiary amines or sterically hindered alkoxides such as tert. alkoxides e.g potassium tert.-butoxide, anion-exchange resins in particular quaternary anion-exchange resins and alkalimetal salts of nuclephilic anions selected in particular from cyanides and fluorides such as potassium cyanide or potassium fluoride. Tertiary amines are preferred in particular diisopropylethyl amine and most preferably triethyl amine.
When a transesterification catalyst is used, its concentration in the mixture catalyst/solvent is generally from 0.5 to 3 mole/liter, preferably from 1 to 2.5 mole/liter and most preferably about 2 mole/liter. In a particular embodiment, the transesterification is carried out with an aminoalcohol, in particular an aminoalcohol having a tertiary amine function and a primary alcohol function such as in particular 2-dimethylaminoethanol. In this embodiment use of supplemental catalyst is not required as the amino function catalyses the reaction. In this embodiment, the use of toxic catalysts such as thallium derivatives as in Savoie et al. Can J. Chem. 1974 p. 2832-9 can be avoided.
In the process according to the invention, the transesterification is generally carried out at a temperature of from 00C to 1000C and preferably from 100C to 600C. A temperature of about 25°C is particularly preferred.
In the process according to the invention, the transesterification is generally carried out at atmospheric pressure.
In the process according to the invention, the free cyclic peptide acid can suitably be obtained from a cyclic peptide ester obtained in step (b) by reaction with a base selected for example from alkaline, quaternary ammonium or earth- alkaline metal hydroxides or carbonates. Particular examples include tetrabutylammonium hydroxide, potassium carbonate and lithium hydroxide. Lithium hydroxide is preferred.
In the process according to the invention, cleavage is generally carried out at a pH of from 8 to 10.
In the process according to the invention, the optional cleavage is generally carried out at a temperature of from 00C to 1000C and preferably from 100C to 600C. A temperature of about 25°C is particularly preferred.
In a preferred embodiment, the invention concerns a process for manufacturing a cyclic peptide having an amide bond formed through intramolecular reaction of a carboxylic group with an amino group which comprises
(a) providing a linear peptide bonded to a Merrifield-type resin comprising a first amino acid having a free amino group and a second amino acid having a free carboxylic group
(b) cyclizing the supported peptide by intramolecular reaction of the carboxylic group with the amino group
(c) cleaving the cyclized supported peptide from the Merrifϊeld type resin by transesterification. In this embodiment, the linear peptide is generally cyclized in the presence of an amide bond forming reagent. The amide bond forming reagent is suitably - A -
selected from coupling reagents for peptide synthesis known in the art such as those described in Bodansky, M., Principles of Peptide Synthesis, 2nd ed. Springer Verlag Berlin/Heidelberg, 1993. Particular examples of amide bond forming agents are selected from mixed anhydrides and other acylating agents such as activated esters or acid halogenides for example isobutyl-chloroformate or pivaloyl chloride. They may be carbodiimides
(e.g. l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide), activated benzotriazine- derivatives (DEPBT : 3-(diethoxyphosphoryloxy)-l,2,3-benzotriazine-4(3H)- one) or uronium or phosphonium salt derivatives of benzotriazol. A preferred amide bond forming reagent is dicylohexylcarbodiimide. It has been found that this cheap reagent gives good results in the cyclization.
In an advantageous embodiment, the process according to the invention further comprises providing a precursor linear peptide having a protected amino group and a protected carboxylic group. In this embodiment the amino group is protected by a first protective group. The first protective group is selected for example from protective groups which can be removed under conditions leaving the bond of the linear peptide to the Merrifϊeld resin substantially intact. Preferred examples of such protective groups include amino protective groups which can be removed by acidolysis selected for example from t.butoxycarbonyl, para-methoxybenzyloxycarbonyl, benzhydryloxycarbonyl, and, preferably, benzyloxycarbonyl. Other examples of such protective groups include amino protective groups which can be removed by reduction with metals or metal compounds such as for example trichloroethoxycarbonyl. In this embodiment, the carboxylic group is protected by a second protective group. The second protective group is selected for example from protective groups which can be removed under conditions leaving the bond of the linear peptide to the Merrifϊeld resin substantially intact. Preferred examples of such protective groups include carboxyl protective groups which can be removed by acidolysis selected for example from para-methoxybenzyl, benzhydryl, and, preferably, t.butyl. Other examples of such protective groups include protective groups which can be removed by reduction with metals or metal compounds such as for example trichloroethyl.
In this embodiment, first and second protective groups are preferably selected to be removable in a single reaction step. Accordingly, the protective groups of the precursor peptide can be removed in a single reaction to provide the linear peptide. Examples of suitable combinations of first and second protective groups which can be removed by acidolysis a single reaction step include benzhydryloxycarbonyl/benzhydryl; para-methoxybenzyloxycarbonyl/ para-methoxybenzyl and, preferably, t.butoxycarbonyl/ t.butyl. Examples of suitable combinations of first and second protective groups which can be removed by reduction with metal or metal compounds in a single reaction step include trichloroethoxycarbonyl/ trichloroethyl
When the removal of protective groups is carried out by acidolysis, suitable reagents are selected for example from perfluoroalkyl carboxylic acids and solutions of HCl in organic solvents preferably polar organic solvents such as dioxane or ethyl acetate. Trifluoroacetic acid is preferred as reagent for acidolysis.
When the removal of protective groups is carried out by reaction with a metal or metal compound, a suitable reagent is, for example, zinc metal. The process according to the invention is particularly suitable when the linear peptide comprises an amino acid having a side chain containing an amino group. Examples of amino acid having a side chain containing an amino group include lysine, ornithine, diaminobutyric acid and diaminopropionic acid. A preferred example of amino acid having a side chain containing an amino group is lysine.
In a particularly preferred embodiment, the linear peptide is bonded to the Merrifield resin through the amino acid having a side chain containing an amino group.
In the process according to the invention the linear peptide can suitably comprise an amino acid having a side chain containing a carboxylic group. Examples of amino acids having a side chain containing a carboxylic group include glutamic acid and aspartic acid. A preferred example is aspartic acid.
In a particularly preferred aspect, the linear peptide comprises both an amino acid having a side chain containing a carboxylic group and an amino acid having a side chain containing an amino group.
The sequence
Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin is more particularly preferred
The side chain carboxylic group or side chain amino group can be suitably protected by the respective protective groups described above.
The invention concerns in particular a process for the manufacture of Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-OH
which comprises providing
Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin, cyclizing the supported peptide by intramolecular reaction of the carboxylic group with the amino group and cleaving the cyclized supported peptide from the Merrifield type resin by transesterifϊcation. The transesterification is generally followed by an operation to cleave the ester such as in particular a saponification to yield the cyclic peptide acid. The invention also concerns a cyclic peptide which is bonded to a
Merrifield-type resin. The supported cyclic peptide according to the invention is preferably as described herein before.
The invention also concerns P-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin I I wherein P is an amino protecting group, in particular an acyl group and preferably an acetyl group (Ac).
The invention also concerns
P-NIe- Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin wherein P is as defined here before.
The example here after is intended to illustrate the invention in a non- limitative manner :
The abbreviations used herein are as follows :
Ac Acetyl
NIe Norleucine
Asp Aspartic acid
His Histidine
Phe Phenylalanine
Arg Arginine
Trp Tryptophan
Lys Lysine
HF Hydrogen Fluoride
Fmoc 9-Fluorenylmethyloxycarbonyl
Boc t-butyloxycarbonyl
RCM resin chloromethyl (Merrifield resin)
SPPS Solid phase peptide synthesis Pbf 2,2,4,6, 7-Pentamethyldihydrobenzofuran-5-sulfony]
Trt trityl tBu t-butyl
TFA trifluoroacetic acid
DI distilled
TIPS triisopropylsilane
DTE dithioerythreitol
DCM dichloromethane
IPA isopropanol
DIEA diisopropylethylamine
DMF dimethylformamide
HOBT hydroxybenzotriazole
DCC dicyclohexylcarbodiimide
TEAP triethylammonium phosphate
LiOH Lithium hydroxide
Example
Sequence : Ac-NIe- Asp-His-D-Phe-Arg-Trp-Lys-OH
Synthesis : 5Og Fmoc-Lys(Boc)-RCM resin (Subst. 0.8 mmol/g) used for synthesis.
Fmoc SPPS strategy used for synthesis at a concentration of 10 ml/gm. The following Amino acid derivatives used for synthesis :
Fmoc-Trp(Boc)-OH
Fmoc-Arg(Pbf)-OH Fmoc-D-Phe-OH
Fmoc-His(Trt)-OH
Fmoc-Asp(OtBu)-OH
Fmoc-Nle-OH
Deprotection Acetylation utilizing Acetic anhydride
Deprotection of peptide on the resin by utilizing the following reagent :
84 % TFA, 5 % Anisole, 5 % DI water, 3 % Thioanisole, 3 % TIPS containing
2 % DTE. Cleaved for 1 to 2 hours.
Washing procedure used after cleavage : 2 x DCM
I x IPA 2 x 5 % DIEA in DCM for 4 minutes each (check pH paper) I x DCM 2 x Methanol 2 x DCM 1 x DMF
Cyclization on the resin by using 5eq. HOBT/ lOeq. DCC in DMF for 48 hours-72 hours.
The Peptide was cleaved from the resin utilizing the following procedure : Peptide resin stirred for 2 days at ambient temperature in 15 % TEA in Methanol. After cleavage from the resin 5 % DI water and 5eq. LiOH added and continued to stir for 2 hours. Acetic acid was added to acidify reaction mixture. Resin filtered off and washed with TFA A Buffer containing 30 % Acetonitrile. Filtrate evaporated or diluted with TFA A Buffer until organic solvent component was below 20 %. Solution filtered and purified. Purification :
TEAP system and then Acetic acid system to yield 5.0486 g pure CS341 (HPLC > 99 %)
Overall yield : 12.3 %

Claims

C L A I M S
1. A process for manufacturing a cyclic peptide which comprises
(a) providing a cyclic peptide bonded to a Merrifield-type resin and
(b) cleaving the cyclic peptide from the Merrifϊeld type resin by transesterification.
2. The process according to claim 1, wherein the transesterification is carried out with a C1-C3 alcohol, preferably methanol.
3. The process according to claim 1, wherein the transesterification is carried out with an alkyl thiol, preferably ethane thiol, or a salt thereof.
4. The process according to any one of claims 1 to 3, wherein the transesterification is carried out at a pH of 8-10
5. The process according to any one of claims 1 to 4 wherein the cyclic structure of the cyclic peptide is formed by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 14, 15, 16, 17, 18, 19 or 20 amino acids.
6. The process according to any one of claims 1 to 5 wherein the cyclic peptide is bonded to the Merrifield-type resin through an amino acid which forms part of the cyclic structure.
7. The process according to any one of claims 1 to 6 wherein the cyclic peptide has an amide bond formed through intramolecular reaction of a carboxylic group with an amino group.
8. The process according to claim 7 wherein the cyclic peptide bonded to the Merrifield resin is provided by
(a) providing a linear peptide bonded to the Merrifield-type resin which linear peptide comprises a first amino acid having a free amino group and a second amino acid having a free carboxylic group and
(b) cyclizing the linear peptide bonded to the Merrifield-type resin by intramolecular reaction of the carboxylic group with the amino group.
9. The process according to claim 8, wherein the linear peptide is cyclized in the presence of an amide bond forming reagent selected from mixed anhydrides, activated esters, acid halogenides, carbodiimides, activated benzotriazine-derivatives and uranium or phosphonium salt derivatives of benzotriazol.
10. The process according to claim 8 or 9, which further comprises providing a precursor linear peptide bonded to the Merrifϊeld-type resin having a protected amino group and a protected carboxylic group.
11. The process according to claim 10, wherein the amino group is protected by a first protective group selected from amino protective groups which can be removed by acidolysis selected for example from t.butoxycarbonyl, para-methoxybenzyloxycarbonyl, benzhydryloxycarbonyl, and, preferably, benzyloxycarbonyl and the carboxylic group is protected by a second protective group selected from carboxyl protective groups which can be removed by acidolysis selected for example from para-methoxybenzyl, benzhydryl, and, preferably, t.butyl.
12. The process according to claim 11, wherein the removal of protective groups is carried out by reaction with trifluoroacetic acid.
13. The process according to claim 10, wherein the amino group is protected by a first protective group selected from amino protective groups which can be removed by reduction with metals or metal compounds and the carboxylic group is protected by a second protective group selected from carboxyl protective groups which can be removed by reduction with metals or metal compounds.
14. The process according to claim 13, wherein the removal of protective groups is carried out by reaction with zinc metal.
15. The process according to claim any one of claims 10 to 14, wherein the protective groups of the precursor linear peptide are removed in a single reaction to provide the linear peptide.
16. The process according to any one of claims 7 to 15, wherein the linear peptide bonded to the Merrifield-type resin comprises an amino acid having a side chain containing an amino group.
17. The process according to claim 16, wherein the amino acid having a side chain containing an amino group is lysine.
18. The process according to claim 16 or 17 wherein the linear peptide is bonded to the Merrifϊeld resin through the amino acid having a side chain containing an amino group.
19. The process according to any one of claims 7 to 18, wherein the linear peptide comprises an amino acid having a side chain containing a carboxylic group.
20. The process according to anyone of claims 1 to 19, wherein the transesterifϊcation is followed by an operation to cleave the ester such as in particular a saponification to yield the cyclic peptide acid.
21. The process according to any one of claims 1 to 20, wherein the cyclic peptide bonded to the Merrifϊeld resin is
Ac-Nle-Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin
22. A cyclic peptide which is bonded to a Merrifield-type resin
23. The cyclic peptide according to claim 22 which is
P-Nle-Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin
wherein P is an amino protecting group.
24. The cyclic peptide according to claim 23 wherein P is acetyl.
25. A linear peptide which is bonded to a Merrifield-type resin, having the formula
P-Nle-Asp-His-D-Phe-Arg-Trp-Lys-Merrifield resin
wherein P is an amino protecting group.
26. The linear peptide according to claim 24 wherein P is acetyl.
PCT/EP2007/057362 2006-07-21 2007-07-17 Process for the manufacture of peptides WO2008009672A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
CN200780034795.6A CN101516903B (en) 2006-07-21 2007-07-17 Process for the manufacture of peptides
EP07787628A EP2049561A1 (en) 2006-07-21 2007-07-17 Process for the manufacture of peptides
US12/374,344 US8415454B2 (en) 2006-07-21 2007-07-17 Process for the manufacture of peptides
JP2009521219A JP5220012B2 (en) 2006-07-21 2007-07-17 Method for producing a peptide

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US83264906P 2006-07-21 2006-07-21
US60/832,649 2006-07-21

Publications (1)

Publication Number Publication Date
WO2008009672A1 true WO2008009672A1 (en) 2008-01-24

Family

ID=38543936

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2007/057362 WO2008009672A1 (en) 2006-07-21 2007-07-17 Process for the manufacture of peptides

Country Status (6)

Country Link
US (1) US8415454B2 (en)
EP (1) EP2049561A1 (en)
JP (1) JP5220012B2 (en)
CN (1) CN101516903B (en)
TW (1) TW200813083A (en)
WO (1) WO2008009672A1 (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101094867B (en) 2004-10-19 2011-08-24 隆萨股份公司 Method for solid phase peptide synthesis
CN106589111B (en) * 2015-10-16 2020-12-29 江苏开元医药化工有限公司 Synthetic method of bremer langdan
CN109306366B (en) * 2017-07-26 2021-12-21 深圳翰宇药业股份有限公司 Method for synthesizing PT141
CN116854805B (en) * 2023-09-05 2023-12-15 杭州湃肽生化科技有限公司 Preparation method of telipopeptide

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993025580A1 (en) * 1992-06-18 1993-12-23 Immunopharmaceutics, Inc. Cyclic peptides that modulate endothelin activity

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0356395A3 (en) * 1988-08-26 1991-09-04 Ciba-Geigy Ag Flanking peptides and process for their preparation
HUT72734A (en) * 1993-04-08 1996-05-28 Ciba Geigy Ag Proces for producing bone-resorption-inhibiting cyclic compounds and compositions
DE19610103A1 (en) * 1996-03-15 1997-09-18 Basf Ag Cycloalkyl derivatives and their synthesis on solid phase
US7250397B2 (en) * 2000-02-23 2007-07-31 Zealand Pharma A/S Antiarrhythmic peptides
AU2003292326A1 (en) * 2002-11-08 2004-06-03 Societe D'extraction Des Principes Actifs (Vincience) Cosmetic or pharmaceutical composition comprising peptides with the sequence arg-gly-ser

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993025580A1 (en) * 1992-06-18 1993-12-23 Immunopharmaceutics, Inc. Cyclic peptides that modulate endothelin activity

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
DAVID FLORA ET AL.: "Detection and control of aspartimide formation in the synthesis of cyclic peptides", BIOORGANIC AND MEDICINAL CHEMISTRY LETTERS, vol. 15, 15 February 2005 (2005-02-15), pages 1065 - 1068, XP002454279 *
E. E. SALEM: "Solid-phase synthesis of a cyclic decapeptide, analog of the antibiotic polymyxin M", DIE PHARMAZIE, vol. 35, no. 12, December 1980 (1980-12-01), pages 761 - 763, XP008084619 *
FAHED AL-OBEIDI ET AL.: "Potent and Prolonged Acting Cyclic Lactam Analogues of alpha-Melanotropin: Design Based on Molecular Dynamics", JOURNAL OF MEDICINAL CHEMISTRY, vol. 32, December 1989 (1989-12-01), pages 2555 - 2561, XP002454278 *
M. T. M. MIRANDA, F. C. THEOBALDO, M. TOMINAGA: "Transesterification of peptide esters and peptidyl resins in methanol-containing calcium acetate", INTERNATIONAL JOURNAL OF PEPTIDE AND PROTEIN RESEARCH, vol. 37, no. 5, 1991, pages 451 - 456, XP008084614 *
MARIA A BEDNAREK ET AL.: "Structure-function studies on the cyclic peptide MT-II, lactam derivative of alpha-melanotropin", PEPTIDES, vol. 20, March 1999 (1999-03-01), pages 401 - 409, XP002454277 *
PETER W. SCHILLER, THI M-D NGUYEN, JACK MILLER: "Synthesis of side-chain to side-chain cyclized peptide analogs on solid supports", INTERNATIONAL JOURNAL OF PEPTIDE AND PROTEIN RESEARCH, vol. 25, 1985, pages 171 - 177, XP008084679 *
See also references of EP2049561A1 *
SYLVIE E BLONDELLE (ED): "Understanding Biology Using Peptides: Proceedinds of the Nineteenth American Peptide Symposium (June 18-23, 2005, San Diego, California)", 23 June 2005, SPRINGER, NEW YORK, XP008084600 *

Also Published As

Publication number Publication date
JP5220012B2 (en) 2013-06-26
US8415454B2 (en) 2013-04-09
JP2009544652A (en) 2009-12-17
CN101516903B (en) 2014-01-01
TW200813083A (en) 2008-03-16
US20090326195A1 (en) 2009-12-31
EP2049561A1 (en) 2009-04-22
CN101516903A (en) 2009-08-26

Similar Documents

Publication Publication Date Title
EP2873677B1 (en) Method of producing self-assembling peptide derivative
US9346850B2 (en) Method of producing peptide
JP5996618B2 (en) Bivalirudine production method
EP3802554B1 (en) Method for solution-phase peptide synthesis and protecting strategies therefore
JP2014517023A (en) Peptide extraction method and its use in liquid phase peptide synthesis
EP3765488A1 (en) Process for the manufacture of pthrp analogue
US8415454B2 (en) Process for the manufacture of peptides
US20220112234A1 (en) Method for producing peptide compound, protective group-forming reagent, and condensed polycyclic compound
WO2017175233A1 (en) Process for large scale liquid phase synthesis of carbetocin and its novel intermediates
EP3781586B1 (en) A method for production of high purity icatibant
US20140099655A1 (en) Methods for assaying enzyme activities
CA2721644C (en) Indolesulfonyl protecting groups for protection of guanidino and amino groups
US20220033440A1 (en) An improved process for the preparation of plecanatide
EP2303914B1 (en) Peptide manufacturing process
US6982315B2 (en) Process for the preparation of carboxamides
JP2008517016A (en) Peptide cyclization on resin
CN116724022A (en) Method for producing peptide, reagent for protecting group formation, and condensed polycyclic compound
CN114621311A (en) Method for synthesizing cyclic heptapeptide Mortiamides and analogues thereof under assistance of DPKO carrier
CN116997559A (en) Method for producing amino acid or peptide, reagent for forming protecting group, and compound
Chen Ionic liquids as novel reaction media for the chemical synthesis of peptides
Carey et al. N α-α, α-dimethyl-3, 5-dialkoxybenzylcarbonyl amino acid 3, 4-dihydro-3-hydroxy-4-oxo-1, 2, 3-benzotriazin-3-yl and pentafluorophenyl esters

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 200780034795.6

Country of ref document: CN

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07787628

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 12374344

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 369/CHENP/2009

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2009521219

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2007787628

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: RU