WO1999064449A2 - Peptide - Google Patents

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Publication number
WO1999064449A2
WO1999064449A2 PCT/GB1999/001848 GB9901848W WO9964449A2 WO 1999064449 A2 WO1999064449 A2 WO 1999064449A2 GB 9901848 W GB9901848 W GB 9901848W WO 9964449 A2 WO9964449 A2 WO 9964449A2
Authority
WO
WIPO (PCT)
Prior art keywords
peptide
positively charged
cell permeable
signal peptide
cell
Prior art date
Application number
PCT/GB1999/001848
Other languages
English (en)
Other versions
WO1999064449A3 (fr
Inventor
John Nelson
Patrick Harriott
Andrew Wallace
Original Assignee
The Queen's University Of Belfast
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
Priority claimed from GBGB9812376.3A external-priority patent/GB9812376D0/en
Priority claimed from GBGB9814888.5A external-priority patent/GB9814888D0/en
Application filed by The Queen's University Of Belfast filed Critical The Queen's University Of Belfast
Priority to EP99955476A priority Critical patent/EP1086126A1/fr
Priority to AU42819/99A priority patent/AU4281999A/en
Publication of WO1999064449A2 publication Critical patent/WO1999064449A2/fr
Publication of WO1999064449A3 publication Critical patent/WO1999064449A3/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • C07K14/003—Peptide-nucleic acids (PNAs)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/50—Fibroblast growth factor [FGF]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C12N15/625—DNA sequences coding for fusion proteins containing a sequence coding for a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]

Definitions

  • the present invention relates to the delivery of molecules into a cell and the use of modified signal peptides.
  • a modified analogue of the signal peptide sequence from Karposi syndrome fibroblast growth factor (kFGF) is used as a cell-permeant vehicle for the intracellular delivery of covalently linked anti-sense peptide nucleic acid sequences (PNAs) .
  • PNAs covalently linked anti-sense peptide nucleic acid sequences
  • PNAs have potential uses as antisense molecules for the control of gene expression. Since they are capable of binding tightly to DNA and RNA targets thus preventing DNA transcription to RNA and RNA translation to protein. These molecules thus have two potential uses of commercial importance:
  • a signal peptide is a short-lived _V-terminal sequence found only on nascent proteins which are synthesised m the endoplasmic reticulum.
  • Signal peptides consist of three domains:
  • Synthetic peptides consisting of only the hydrophobic cores are typically insoluble m water.
  • the signal peptide sequence of Karposi syndrome-derived FGF as an example, we have modified these insoluble sequences by the addition of positively charged ammo acids (for example lysmes) , which have the effect of rendering them water soluble without compromising their ability to translocate across cellular membranes.
  • ammo acids for example lysmes
  • a cell permeable peptide comprising at least the hydrophobic core of a signal peptide or an analogue thereof wherein the peptide is modified by the addition of at least one positively charged amino acids or positively charged analogues thereof .
  • the signal peptide may be a natural or synthetic signal peptide or a peptide which is substantially similar thereto.
  • a peptide which is substantially similar to a signal peptide is at least 60% homologous thereto.
  • At least one positively charged amino acid is chosen from lysine and/or arginine and/or any positively charged analogues thereof.
  • the cell permeable peptide is a modified analogue of Karposi syndrome fibroblast growth factor (kFGF) .
  • kFGF Karposi syndrome fibroblast growth factor
  • the positively charged amino acid consists of one or more lysine residues.
  • the invention further provides the use of cell permeable peptides as described herein for intracellular delivery of a molecule.
  • one or more lysine residues will be attached to the C terminal of the signal sequence peptide or signal sequence peptide analogue.
  • This positively charged lysine allows the linkage of a peptide nucleic acid, thus facilitating m vivo delivery of the said peptide nucleic acid.
  • the invention also provides a cell permeable peptide which contains multiple positively charged ammo acids or positively charged analogues thereof wherein a peptide nucleic acid may be conjugated to each positively charged residue and wherein the peptide nucleic acids conjugated by such a means are identical or different.
  • the invention also provides a cell permeable peptide which comprises at least one positively charged ammo acid residue or functionally equivalent positively charged analogue thereof conjugated or conjugatable to a lysine tree, to which multiple peptide nucleic acids may be joined for transport and presentation.
  • the linked peptide nucleic acid sequence may be antisense.
  • the peptide nucleic acid sequence will be covalently linked.
  • the present invention thus allows the use of cell permeable peptides as described herein to deliver peptide nucleic acids to ln-vivo targets.
  • PNAs Polymethyl methacrylate-N-(2-ammoethyl)-N-(2-ammoethyl) glycme units to which natural nucleobases are attached through methylenecarbonyl linkers.
  • N- (2-ammoethyl) glycme units to which natural nucleobases are attached through methylenecarbonyl linkers.
  • PNAs suffer from similar accessibility problems as phosphorothioates do, and passive diffusion of unmodified PNA across lipid membranes is not efficient ( ittung, P., et al . , 1995) .
  • a small number of native peptide sequences can translocate across membranes of living cells in an energy- independent and receptor- independent manner. These peptides have been used to import active cargo into the cell. For example a peptide from the homeodomain of Antennapedia has been successfully used to import both peptidal inhibitors of protein kinase C (Theodore, et al . , 1995) and conventional anti-sense oligonucleotides (Allinquant, et al . , 1995) .
  • the present invention provides use of cell permeable peptide import (CPPI) to deliver peptide nucleic acids (PNAs) .
  • CPPI cell permeable peptide import
  • PNAs peptide nucleic acids
  • the present invention provides use of the signal peptide sequence from Karposi syndrome fibroblast growth factor (kFGF) for delivery of antisense peptide nucleic acid sequences (PNAs) .
  • kFGF Karposi syndrome fibroblast growth factor
  • PNAs antisense peptide nucleic acid sequences
  • the invention provides use of a peptide as defined herein together with lysine residues for multiple presentation of peptide nucleic acids.
  • the invention further provides use of peptides as defined herein together with lysine residues in the simultaneous presentation of different peptides nucleic acids.
  • the present invention combines the two above technologies to use CPPI to deliver PNAs to in vi vo targets .
  • the modified signal peptides described m this invention can be used for the delivery of any cell-impermeant substance into cells.
  • the signal peptides described m this invention can be used to improve the delivery of substances of low permeability into cells.
  • the signal peptide delivery system has commercial value m therapeutic drug-delivery systems including, but not restricted to, gene therapy, cancer therapy and anti-mfectious agent therapy.
  • This system also has commercial value as a tool for biochemical and molecular biological research.
  • Figure 1 illustrates carboxyfluorescein labelled kFGF signal peptide-Lys .Lys .Lys - fluoresence calibration curve .
  • Figure 2 illustrates carboxfluorescein labelled cell permeant peptide incorporation by whole human endothelial cells.
  • Figure 3 depicts incorporation of carboxyfluorescein labelled signal peptide-Lys . Lys . Lys by cell.
  • Figure 4 illustrates subcellular distribution of labelled signal peptide in cells.
  • Figure 5 depicts incorporation of labelled kFGF peptide into human dermal endothelial cells.
  • Figure 6a sets out the signal peptide sequence and modifications.
  • Figure 6b illustrates simultaneous presentation of 3 PNAs directed to different sites on a target RNA.
  • Figure 6c illustrates multiple presentation of the single PNA species.
  • Table 1 describes carboxyfluorescein derivatised cell permeant peptides.
  • Table 2a sets out uptake of cell permeant peptides by cells.
  • Table 2b sets out cellular uptake of permeant peptides by BHK cells.
  • Table 3 sets out results of washing labelled antennapedia cells.
  • Table 4 sets out washing results for labelled signal peptide-KKK and cells.
  • the raw relative fluorescent units (RFU) values were converted to nMoles per 10 6 cells using a calibration curve constructed for each peptide.
  • An example of a fluorescence calibration curve of fluorescein labelled kFGF is shown in Figure 1.
  • the kFGF-KKK sequence shows similar high rates of cytosolic and nuclear incorporation compared with the antennapedia peptide (Table 2A) .
  • the PKC and substance P peptides show much lower incorporation Table 2A & 2B) .
  • Incorporation of the kFGF-KKK sequence is saturable, as can be seen from the data presented on Figure 2 and time-dependent as shown in Figure 3.
  • Table 2A shows that antennapedia is lost during subcellular fractionation. Unlike the antennapedia peptide, carboxyfluorescein-kFGF signal peptide-KKK is not loosely attached to the cell surface as shown in Tables 3 and 4. Unlike the antennapedia peptide, carboxyfluorescein-kFGF signal peptide-KKK does not remain membrane -bound as shown by the data presented in Figure 4.
  • oligonucleotide sequences or those in which the phosphodiester bonds are replaced with nuclease-resistant bonds may be conjugated to the kFGF-derived delivery system for intracellular delivery and subsequent specific blocking of gene translation or Rnase-targeted destruction of the mRNA in question.
  • nuclease-resistant bonds such as the phosphothiorates and the like
  • peptide nucleic acid sequences may be used, as m example 1.
  • PNA Peptide Nucleic Acid
  • Nuclear localisation signal (NLS) sequences such as are found on transcription factors like NF-kappaB may be conjugated to the kFGF-derived delivery system, as m Example 1. Intracellular delivery of NLS peptide sequences would act as 'bait' to selectively block the translocation of the selected transcription factor, thus preventing its action. In this way, genes under the control of the transcription factor could be identified on the basis of down regulated expression.
  • NLS Nuclear localisation signal
  • Signal transduction motifs such as phosphotyros e- containing peptide sequences (pYP's) act as docking sites for a large number of proteins.
  • Such signalling proteins contain domains that recognise (contextually) the phosphotyrosme residues and bind to them m a specific manner.
  • pYP's are recognised by SH-2 (Src- homology-2) domains and PTB (phosphotyrosme binding domains) .
  • Specificity is provided by short ammo acid sequences iV-and/or C-termmal of the phosphotyrosme.
  • Such peptide motifs could be conjugated to the kFGF peptide-de ⁇ ved delivery system as m Example 1, and could be used to mtracellularly deliver pYP's which would act as bait, thus allowing signal pathways to be 'interrogated'.
  • FIG. 6A The signal sequence of kFGF was modified to contain three lysmes at the C-termmal of the hydrophobic signal sequence. This procedure is illustrated m Figure 6A.
  • Figure 6A (I) shows the signal peptide with an attached reporter group.
  • Figure 6A Part II illustrates the addition of the t ⁇ -lysme extension to the C-termmal of the signal peptide sequence, thus providing three positive charges which aid solubility and cell permeability.
  • Figure 6A Part Illb the peptide nucleic acid forms part of the linear primary ammo acid sequence, with Part IV illustrating a t ⁇ -lysme C-termmal extension to the peptide nucleic acid sequence providing 3 positive charges and aiding solubility and cell permeability.
  • Part V of Figure 6A further shows a tri-lysyl extension at the N-termmal of the signal peptide which provides 3 positive charges aiding solubility and cell permeability.
  • the addition of the tri-lysyl extension proximal to the carboxyfluorescein reporter group enhances its fluorescence.
  • the peptide nucleic acid sequence initially forms part of the linear primary ammo acid sequence at the N- terminal of the original peptide, before a tri-lysyl extension is added to the N-termmal of the peptide nucleic acid extension.
  • This peptide therefore, can accommodate three PNAs, each bonded to a lysine epsilon amino group.
  • This can be extended using the Multiple Antigen Presentation (MAP) technology to present eight (or more) PNA' s on one kFGF signal sequence.
  • MAP Multiple Antigen Presentation
  • a 'lysine tree' constructed in this way accommodates eight copies of the same PNA, thus increasing the effective concentration delivered by each CPPI.
  • Part I An example of the addition of such a lysine tree is shown in Figure 6C Parts I -IV.
  • Part I a single lysine molecule added to the C-terminal of the kFGF signal peptide sequence allows the multiple PNA lysine tree to be added to the e-amino group of the lysine side chain.
  • a lysine molecule added to the N-terminal of the kFGF signal peptide sequence allows the multiple PNA lysine tree to be added to the e-amino group of the lysine side chain.
  • Part III of Figure 6C further shows that when a C- terminal tri-lysine extension is added to the signal peptide with N-terminal associated multiple PNA lysine tree, the 3 positive charges aid solubility and cell permeability of the molecule.
  • Part IV of Figure 6C add a tri-lysyl extension at the N-terminal of the signal peptide which is attached to the lysine group added to allow attachment of the multiple PNA lysine tree as originally illustrated in Figure 6C Part II.
  • the addition of the 3 positively charged molecules at this terminal of the molecule, proximal to the carboxyfluorescein reporter group enhances its fluorescence.
  • a carrier can be constructed containing three (or more) different PNAs directed towards different sites on the same target mRNA. This strategy has been termed 'molecular triangulation' (Branch, A.D. , 1998) .
  • Figure 6B illustrates this process of 'molecular triangulation' .
  • Figure 6B Part I shows the signal peptide with a C-terminal tri-lysyl extension which allows three different PNA sequences to be conjugated to the epsilon-amino groups of the three lysines.
  • Figure 6B Part III shows the addition of a further three lysines to the molecule of Part I, which adds three positive charges, which aid solubility and cell permeability.
  • Figure 6B Part III shows the addition of the tri-lysyl extension to the N-terminal of the molecule of Part I. Again the 3 positive charges aid the solubility and cell permeability of the molecule, which their proximal location to the carboxyfluorescein reporter group enhances its fluorescence.
  • Figure 6B Part IV, illustrates an N-terminal tri-lysyl extension added to the kFGF signal peptide sequence, which subsequently allows three different PNA sequences to be conjugated to the epsilon-amino groups of the lysines. Further, this molecule has 3 lysines added at the C- terminal to add positive charge which aid solubility and cell permeability.
  • Figure 6B Part V shows the signal peptide again with the three peptide nucleic acid associated tri-lysine extension at the N-terminal, but with the addition of the further 3 lysine groups also being made to the N-terminal where they will have the effect of aiding solubility and cell permeability, which also enhance the fluorescence of the carboxyfluorescein reporter group due to their proximity.
  • Lysine extensions comprising more or less than three lysine residues may also be useful to provide additional solubility and cell permeability.
  • the lysine extension may be provided next to a carboxyfluorescein reporter group to enhance its fluorescence.

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Abstract

La présente invention concerne un nouveau procédé d'apport de molécules à l'intérieur d'une cellule dans lequel on utilise un peptide signal modifié lié à un acide nucléique peptidique. Le peptide signal comprend au moins un résidu d'acide aminé de charge positive ou un équivalent fonctionnel d'un tel résidu. Ces résidus à charge positive ajoutés servent de groupe de liaison permettant la fixation des acides nucléiques peptidiques sur le peptide signal, augmentant de la sorte le nombre de séquences nucléotidiques peptidiques délivrées par le peptide signal et renforçant, par conséquent, l'efficacité fonctionnelle de ce dernier. L'extension du peptide signal à l'extrémité C ou N-terminale via l'adjonction d'un résidu à charge simple ou multiple ou d'analogues de ce dernier permet de modifier et d'améliorer la fonction d'apport du peptide signal en augmentant les caractéristiques de solubilité et de perméabilité cellulaire de celui-ci.
PCT/GB1999/001848 1998-06-10 1999-06-10 Peptide WO1999064449A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP99955476A EP1086126A1 (fr) 1998-06-10 1999-06-10 Peptide permeable aux cellules
AU42819/99A AU4281999A (en) 1998-06-10 1999-06-10 Peptide

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB9812376.3 1998-06-10
GBGB9812376.3A GB9812376D0 (fr) 1998-06-10 1998-06-10
GB9814888.5 1998-07-10
GBGB9814888.5A GB9814888D0 (fr) 1998-07-10 1998-07-10

Publications (2)

Publication Number Publication Date
WO1999064449A2 true WO1999064449A2 (fr) 1999-12-16
WO1999064449A3 WO1999064449A3 (fr) 2002-10-24

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PCT/GB1999/001848 WO1999064449A2 (fr) 1998-06-10 1999-06-10 Peptide

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EP (1) EP1086126A1 (fr)
AU (1) AU4281999A (fr)
WO (1) WO1999064449A2 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001027154A2 (fr) * 1999-09-27 2001-04-19 Mahony Daniel J O Systeme de relargage de medicaments a base d'un peptide de translocation membranaire
WO2001057072A2 (fr) * 2000-02-07 2001-08-09 Wisconsin Alumni Research Foundation Peptides antiviraux pharmacologiquement actifs et leurs methodes d'utilisation
WO2002055545A1 (fr) * 2001-01-12 2002-07-18 Rhobio Pseudopeptides antimicrobiens
EP1339431A1 (fr) * 2000-11-30 2003-09-03 Unigene Laboratories, Inc. Apport oral ameliore de peptides au moyen de translocateurs de membrane pouvant etre coupes par une enzyme
GB2346616B (en) * 1998-11-13 2004-04-21 Cyclacel Ltd Transport vectors
EP1862471A2 (fr) * 2000-02-07 2007-12-05 Wisconsin Alumni Research Foundation Peptides antiviraux pharmacologiquement actifs et leurs procédés d'utilisation
US7316819B2 (en) 2001-03-08 2008-01-08 Unigene Laboratories, Inc. Oral peptide pharmaceutical dosage form and method of production
US7432045B2 (en) 2003-12-01 2008-10-07 Wisconsin Alumni Research Foundation Method of inhibiting influenza infection with antiviral peptides
US8017727B2 (en) 1999-09-27 2011-09-13 Merrion Research Iii Limited Conjugates of membrane translocating agents and pharmaceutically active agents
US8088734B2 (en) 2003-01-21 2012-01-03 Unigene Laboratories Inc. Oral delivery of peptides
US8835377B2 (en) 2004-06-18 2014-09-16 Ugp Therapeutics, Inc. Oral delivery of peptide pharmaceutical compositions
US20150119340A1 (en) * 2013-10-29 2015-04-30 Samsung Electronics Co., Ltd. Fusion peptide and use thereof for cell membrane penetrating
US10072065B2 (en) 2015-08-24 2018-09-11 Mayo Foundation For Medical Education And Research Peptide-mediated delivery of immunoglobulins across the blood-brain barrier
US10377813B2 (en) 2009-07-14 2019-08-13 Mayo Foundation For Medical Education And Research Peptide-mediated non-covalent delivery of active agents across the blood-brain barrier
US10421967B2 (en) 2014-05-15 2019-09-24 Hoffmann-La Roche Inc. Oligomers and oligomer conjugates

Citations (1)

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WO1997004006A1 (fr) * 1995-07-18 1997-02-06 Friedhelm Herrmann Substance utilisee pour lutter contre la croissance tumorale et les infections virales

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WO1997004006A1 (fr) * 1995-07-18 1997-02-06 Friedhelm Herrmann Substance utilisee pour lutter contre la croissance tumorale et les infections virales

Non-Patent Citations (5)

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Title
ALLINQUANT B ET AL., : "Downregulation of amyloid precursor protein inhibits neurite outgrowth in vitro" THE JOURNAL OF CELL BIOLOGY, vol. 128, no. 5, March 1995 (1995-03), pages 919-927, XP000856389 cited in the application *
DU C ET AL., : "Conformational and topological requirements of cell-permeable peptide function" JOURNAL OF PEPTIDE RESEARCH, vol. 51 (3), March 1998 (1998-03), page 235-243 XP000856550 *
FULLER-PACE F ET AL., : "Cell transformation by kappaFGF requires secretion but not glycosylation" J. CELL. BIOL. , vol. 115, no. 2, 1991, pages 547-555, XP000856406 *
LIN Y-Z ET AL., : "Inhibition of nuclear translocation of transcription factor NF-kappaB by a synthetic peptide containing a cell membrane-permeable motif and nuclear localization sequence" JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 270, no. 24, 1995, pages 14255-14258, XP002050723 *
ROJAS M. ET AL., : "Controlling epidermal growth factor (EGF)-stimulated Ras activation in intact cells by a cell-permeable peptide mimicking phosphorylated EGF receptor" JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 271, no. 44, 1996, page 27456-27461 XP002124151 *

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2346616B (en) * 1998-11-13 2004-04-21 Cyclacel Ltd Transport vectors
US8017727B2 (en) 1999-09-27 2011-09-13 Merrion Research Iii Limited Conjugates of membrane translocating agents and pharmaceutically active agents
WO2001027154A3 (fr) * 1999-09-27 2002-06-20 Daniel J O'mahony Systeme de relargage de medicaments a base d'un peptide de translocation membranaire
WO2001027154A2 (fr) * 1999-09-27 2001-04-19 Mahony Daniel J O Systeme de relargage de medicaments a base d'un peptide de translocation membranaire
US7371809B2 (en) 2000-02-07 2008-05-13 Wisconsin Alumni Research Foundation Pharmacologically active antiviral peptides
WO2001057072A2 (fr) * 2000-02-07 2001-08-09 Wisconsin Alumni Research Foundation Peptides antiviraux pharmacologiquement actifs et leurs methodes d'utilisation
JP2003522185A (ja) * 2000-02-07 2003-07-22 ウイスコンシン アラムニ リサーチ ファンデーション 薬理活性抗ウイルスペプチドおよびそれらの使用法
US8748566B2 (en) 2000-02-07 2014-06-10 Wisconsin Alumni Research Foundation Pharmacologically active antiviral peptides and methods of use
EP1862471A2 (fr) * 2000-02-07 2007-12-05 Wisconsin Alumni Research Foundation Peptides antiviraux pharmacologiquement actifs et leurs procédés d'utilisation
JP4896333B2 (ja) * 2000-02-07 2012-03-14 ウイスコンシン アラムニ リサーチ ファンデーション 薬理活性抗ウイルスペプチドおよびそれらの使用法
US8748565B2 (en) 2000-02-07 2014-06-10 Wisconsin Alumni Reseach Foundation Pharmacologically active antiviral peptides and methods of their use
EP1862471A3 (fr) * 2000-02-07 2008-05-28 Wisconsin Alumni Research Foundation Peptides antiviraux pharmacologiquement actifs et leurs procédés d'utilisation
WO2001057072A3 (fr) * 2000-02-07 2002-10-31 Wisconsin Alumni Res Found Peptides antiviraux pharmacologiquement actifs et leurs methodes d'utilisation
EP1339431A1 (fr) * 2000-11-30 2003-09-03 Unigene Laboratories, Inc. Apport oral ameliore de peptides au moyen de translocateurs de membrane pouvant etre coupes par une enzyme
EP1339431A4 (fr) * 2000-11-30 2005-10-12 Unigene Lab Inc Apport oral ameliore de peptides au moyen de translocateurs de membrane pouvant etre coupes par une enzyme
WO2002055545A1 (fr) * 2001-01-12 2002-07-18 Rhobio Pseudopeptides antimicrobiens
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AU4281999A (en) 1999-12-30
WO1999064449A3 (fr) 2002-10-24
EP1086126A1 (fr) 2001-03-28

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