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WO1993005065A1 - Preparation de peptides par une synthese en phase solide, et intermediaires pour la preparation - Google Patents

Preparation de peptides par une synthese en phase solide, et intermediaires pour la preparation Download PDF

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Publication number
WO1993005065A1
WO1993005065A1 PCT/GB1992/001567 GB9201567W WO9305065A1 WO 1993005065 A1 WO1993005065 A1 WO 1993005065A1 GB 9201567 W GB9201567 W GB 9201567W WO 9305065 A1 WO9305065 A1 WO 9305065A1
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WO
WIPO (PCT)
Prior art keywords
compound
general formula
group
solid support
amino acid
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.)
Ceased
Application number
PCT/GB1992/001567
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English (en)
Inventor
Ram Prakash Sharma
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.)
PORTON DEVELOPMENTS Ltd
Original Assignee
PORTON DEVELOPMENTS Ltd
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 PORTON DEVELOPMENTS Ltd filed Critical PORTON DEVELOPMENTS Ltd
Priority to EP92918056A priority Critical patent/EP0600996A1/fr
Publication of WO1993005065A1 publication Critical patent/WO1993005065A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/04—Esters of silicic acids
    • 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
    • C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
    • C07K1/061—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using protecting groups
    • C07K1/062—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using protecting groups for alpha- or omega-carboxy functions
    • 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
    • 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
    • C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
    • C07K1/08—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents
    • C07K1/088—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents containing other elements, e.g. B, Si, As

Definitions

  • the present invention relates to the preparation of
  • a tertiary butyl oxycarbonyl group is a typical amine protecting group, for example tert-butyl oxycarbonyl (tBoc), which may be removed either by hydrochloric acid gas in organic solvents or by using trifluoroacetic acid (TFA).
  • tBoc tert-butyl oxycarbonyl
  • trialkylsilyl protecting groups disclosed in this prior art document include trimethylsilyl- (TMS) and
  • tert-butyldimethylsilyl- (TBDMS) esters of amino acids it is essential to employ a temporary N-protecting group, for example tBoc, to prevent N-silyl derivatisation.
  • tBoc temporary N-protecting group
  • the subsequent removal of the N-protection with acid yields the amino acid trialkylsilyl ester acid salt, which must be neutralised before the amino group can participate in peptide bond formation.
  • the preparation of these amino acid trialkylsilyl esters is thus a three-step process.
  • An example for the production of an amino acid TMS-ester is shown below: tBoc. NH . CHR. COOH + TMSC1 + Base tBoc . NH . CHR. COOTMS . + Base-HC1
  • TBDMS-esters are used in solid-phase peptide synthesis.
  • This activation can be difficult to achieve and an accurate measure of the amount of amino acid bound to the resin is not easy to obtain.
  • this type of linkage is very stable and requires unfavourably harsh acidic conditions, such as liquid HF, for cleavage of the synthesised peptide.
  • N ⁇ -amino groups is known per se in conventional synthetic chemistry (e.g. cf. CARPINO, L.A. & HAN, G.Y. (1972) J. Org. Chem 37 3404) but harsh conditions have precluded the use of such linker compounds in peptide syntheses.
  • the present invention provides a process for the production of a peptide of general formula (I) (I)
  • the present invention provides a process for the production of a peptide of general formula (I) (I)
  • linker compounds with phosgene.
  • linkers may also be attached to any solid support W bearing a terminal amino function.
  • A) allows release of a newly synthesised peptide under mild conditions such as 15% piperidine in dimethyl formamide.
  • B) the conditions for peptide release are slightly more rigorous, requiring 90% trifluoroacetic acid (TFA).
  • Cleavage from C) can be achieved under mild basic conditions, such as 0.1N sodium hydroxide.
  • the solid support W is a resin having a terminal amino group, for example commercially available resins MBHA., BHA., or aminomethyl ated resins.
  • W and the linker compound are preferably reacted to form a W-Y bond in the presence of DCC/HOBt.
  • linker group containing solid supports W-Y for example:
  • Steps (c) or (d) and (d) or (e), respectively) can take place simultaneously.
  • Any reactive side chains on residues A are protected and subsequently deprotected, as necessary by known procedures.
  • the choice of side-chain protecting groups may be adjusted to suit the method of removal of the protecting group Z.
  • the invention further provides compounds of general formula (II); (II)
  • A is either Asp or Glu
  • Figure 1 is an HPL Chromatograph of Leucine-Enkephalin synthesised using tert-butoxysilyl (TBOS) esters;
  • Figure 2 is a FIB Mass Spec Analysis of Leu-Enkephalin synthesised using L-amino acid-TBOS esters;
  • Figure 3 is an HPL Chromatograph of Leucine Enkephalin Alcohol
  • Figure 4 is a FIB Mass Spec of Leucine Enkephalin Alcohol
  • Figure 5 is a FIB Mass Spec of Leucine Enkephalin Diol.
  • Figure 6 is a FIB Mass Spec of Leucine Enkephalin Chloromethyl Ketone. The invention will now be illustrated by way of example with reference to the following description.
  • This compound may be produced by a process which comprises the following steps:
  • a synthesis can be carried out using compound (9) according to the following procedure:
  • Benzyl acetate resin Chioromethylated co-polystyrene-2% divinylbenzene (5g. 1mmol./g.) suspended in 2-methoxyethanol
  • the resin was then swollen in dimethyl acetamide (2 x 10mls.) and treated with L-tryosine methyl ester hydrochloride (1.0mmol.; 0.231g.) and triethylamine (2.0mmol.; 0.112mls.) in
  • the leucine-enkephalin pentapeptide was assembled by four successive cycles of addition and deprotection to yield the desired peptidyl-resin.
  • a typical addition cycle consisted of treating the deprotected resin with the amino acid TBOS ester (1.0mmol.), HOBt (1.0mmol; 0.135g.) and diisoproplycarbodiimide (1.0mmol.; 0.156mls.) in DCM (10mls.) for 4 hours at room temperature. The resin would then be washed thoroughly with DCM before being deprotected by treatment with 25% TFA in DCM (10mls.) for 10 minutes at room temperature followed by washing with DCM (6 x 10mls.) before commencing the subsequent addition cycle.
  • Buffer A 0.1% TFA., H 2 O
  • the preferred carboxy-protected amino acids (II) are the tri-t-butoxysilyl esters. These can easily be produced in a one step process as follows:
  • the reaction is rapid and typically yields 85-90% product.
  • the simplicity of production contrasts favourably with the prior three step process for production of the silyl esters of W090/05738.
  • the tri-t-alkoxysilyl esters, alkyl-di-t-alkoxysilyl esters and dialkyl-t-alkoxysilyl esters can be made from free amino acids without the need for N ⁇ -protection (by groups such as tBoc).
  • the free amino acids are abundantly available and cheap.
  • tri-t-alkoxysilyl esters, alkyl-di-t-alkoxysilyl esters and di alkyl-t-alkoxysilyl esters show good stability when stored under anhydrous conditions, hence permitting long term storage. Large scale preparation of these derivatives may be carried out well in advance of the peptide synthesis.
  • esters are stable between pH 4.0 and pH 8.0 and are stable to alcoholic solvents. They can be readily removed at pH values less than 4 or greater than 8.
  • the procedure for the preparation of several tri-t-butoxysilyl amino acid esters is illustrated in more detail in the example below:
  • the pyridinium hydrochloride is removed by filtration, washing the precipitate with ethyl acetate (50ml). All solvents, (i.e.
  • the product was characterised by means of its infra-red (I.R.) spectrum, thin layer chromatography (t.l.c.) and Mass Spectrum (M.S.).
  • I.R. infra-red
  • t.l.c. thin layer chromatography
  • M.S. Mass Spectrum
  • the mild nature of the synthetic strategy proposed may provide a means of synthesising modified peptides, such as phosphopeptides or glycopeptides, that cannot withstand the conditions used at some stages of conventional synthetic procedures.
  • modified peptides such as phosphopeptides or glycopeptides
  • the reversal of the direction of synthesis from the conventional C- to -N terminal also opens up the possibility of carrying out solid-phase fragment coupling for the production of large peptides.
  • a further advantage of synthesising in the N to C direction is that the C-terminal remains free and this allows for modifying the C-terminal of peptides whilst they are bound on the solid support.
  • Examples of C-terminal modified peptides which have been prepared are enkephalin-alcohol, -diol and -chloromethylketone. Data on these analogues is shown in Figures 3-6. The products made
  • t.l.c solvent was chloroform: methanol (9:1).
  • Bz refers to Benzyl.

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  • 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

L'invention se rapporte à un procédé de préparation de polypeptides par synthèse en phase solide. Ledit procédé consiste à fixer des acides aminés protégés par carboxy à l'extrémité carboxyle d'une chaîne peptidique liée à son extrémité N-terminale à un support solide. La protection carboxy est faite par un groupe tertiaire alcoxy silyle. L'invention se rapporte également à de nouveaux esters d'acides aminés du groupe tertiaire alcoxy silyle pouvant être utilisés comme intermédiaires.
PCT/GB1992/001567 1991-08-30 1992-08-26 Preparation de peptides par une synthese en phase solide, et intermediaires pour la preparation Ceased WO1993005065A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP92918056A EP0600996A1 (fr) 1991-08-30 1992-08-26 Preparation de peptides par une synthese en phase solide, et intermediaires pour la preparation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9118669.2 1991-08-30
GB919118669A GB9118669D0 (en) 1991-08-30 1991-08-30 Preparation of peptides by a soliphase synthesis and intermediates therefor

Publications (1)

Publication Number Publication Date
WO1993005065A1 true WO1993005065A1 (fr) 1993-03-18

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1992/001567 Ceased WO1993005065A1 (fr) 1991-08-30 1992-08-26 Preparation de peptides par une synthese en phase solide, et intermediaires pour la preparation

Country Status (4)

Country Link
EP (1) EP0600996A1 (fr)
AU (1) AU2464992A (fr)
GB (1) GB9118669D0 (fr)
WO (1) WO1993005065A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5798035A (en) * 1996-10-03 1998-08-25 Pharmacopeia, Inc. High throughput solid phase chemical synthesis utilizing thin cylindrical reaction vessels useable for biological assay
WO2002094857A1 (fr) * 2001-05-23 2002-11-28 The Curators Of The University Of Missouri Synthese peptidique en phase solide inverse
WO2006097698A1 (fr) * 2005-03-14 2006-09-21 Activotec Spp Limited Synthese de peptide en phase solide inversee avec une etape de coiffage supplementaire
US7786259B2 (en) 2001-05-23 2010-08-31 The Curators Of The University Of Missouri Attachment and elaboration strategies for inverse peptide synthesis
USRE46830E1 (en) 2004-10-19 2018-05-08 Polypeptide Laboratories Holding (Ppl) Ab Method for solid phase peptide synthesis
WO2019069978A1 (fr) 2017-10-03 2019-04-11 日産化学株式会社 Procédé de production de composé peptidique
US20220306682A1 (en) * 2019-08-30 2022-09-29 Nissan Chemical Corporation Method for producing peptide compound

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005738A1 (fr) * 1988-11-19 1990-05-31 Public Health Laboratory Service Board Trialkylsilyle esters d'acides amines et leur emploi dans la synthese de peptides
DE3924257A1 (de) * 1989-07-19 1991-01-31 Gruszecki Wojciech Silikonester als neue schutzgruppen in der organischen chemie

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005738A1 (fr) * 1988-11-19 1990-05-31 Public Health Laboratory Service Board Trialkylsilyle esters d'acides amines et leur emploi dans la synthese de peptides
DE3924257A1 (de) * 1989-07-19 1991-01-31 Gruszecki Wojciech Silikonester als neue schutzgruppen in der organischen chemie

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 115, no. 19, 11 November 1991, Columbus, Ohio, US; abstract no. 208480g, W GRUSZECKI ET AL. 'stable tri-t-butoxysilyl esters of aspartic and glutamic acid as aa new protection of their carboxyl groups' page 1076 ;column RIGHT ; *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5798035A (en) * 1996-10-03 1998-08-25 Pharmacopeia, Inc. High throughput solid phase chemical synthesis utilizing thin cylindrical reaction vessels useable for biological assay
USRE37194E1 (en) 1996-10-03 2001-05-29 Pharmacopeia, Inc. High throughput solid phase chemical synthesis utilizing thin cylindrical reaction vessels useable for biological assay
WO2002094857A1 (fr) * 2001-05-23 2002-11-28 The Curators Of The University Of Missouri Synthese peptidique en phase solide inverse
US7214769B2 (en) 2001-05-23 2007-05-08 The Curators Of The University Of Missouri Method for inverse solid phase synthesis of peptides
US7786259B2 (en) 2001-05-23 2010-08-31 The Curators Of The University Of Missouri Attachment and elaboration strategies for inverse peptide synthesis
USRE46830E1 (en) 2004-10-19 2018-05-08 Polypeptide Laboratories Holding (Ppl) Ab Method for solid phase peptide synthesis
WO2006097698A1 (fr) * 2005-03-14 2006-09-21 Activotec Spp Limited Synthese de peptide en phase solide inversee avec une etape de coiffage supplementaire
GB2437901A (en) * 2005-03-14 2007-11-07 Activotec Spp Ltd Inverse solid phase peptide synthesis with additional capping step
WO2019069978A1 (fr) 2017-10-03 2019-04-11 日産化学株式会社 Procédé de production de composé peptidique
CN111164092A (zh) * 2017-10-03 2020-05-15 日产化学株式会社 肽化合物的制造方法
US20200291061A1 (en) * 2017-10-03 2020-09-17 Nissan Chemical Corporation Method for producing peptide compound
JPWO2019069978A1 (ja) * 2017-10-03 2020-11-05 日産化学株式会社 ペプチド化合物の製造方法
EP3693377A4 (fr) * 2017-10-03 2021-06-30 Nissan Chemical Corporation Procédé de production de composé peptidique
JP7196087B2 (ja) 2017-10-03 2022-12-26 日産化学株式会社 ペプチド化合物の製造方法
TWI811246B (zh) * 2017-10-03 2023-08-11 日商日產化學股份有限公司 胜肽化合物之製造方法
CN111164092B (zh) * 2017-10-03 2023-12-26 日产化学株式会社 肽化合物的制造方法
US20220306682A1 (en) * 2019-08-30 2022-09-29 Nissan Chemical Corporation Method for producing peptide compound
US12466850B2 (en) * 2019-08-30 2025-11-11 Nissan Chemical Corporation Method for producing peptide compound

Also Published As

Publication number Publication date
GB9118669D0 (en) 1991-10-16
AU2464992A (en) 1993-04-05
EP0600996A1 (fr) 1994-06-15

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