WO1997004006A1 - Substance for use against tumour growth and viral infections - Google Patents

Substance for use against tumour growth and viral infections Download PDF

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Publication number
WO1997004006A1
WO1997004006A1 PCT/DE1996/001286 DE9601286W WO9704006A1 WO 1997004006 A1 WO1997004006 A1 WO 1997004006A1 DE 9601286 W DE9601286 W DE 9601286W WO 9704006 A1 WO9704006 A1 WO 9704006A1
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cell
sequence
bcl
peptide
permeable peptide
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PCT/DE1996/001286
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German (de)
French (fr)
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Friedhelm Herrmann
Marion Brach
Michael Kiehntopf
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Friedhelm Herrmann
Marion Brach
Michael Kiehntopf
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Publication of WO1997004006A1 publication Critical patent/WO1997004006A1/en

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    • 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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence

Definitions

  • the invention relates to an agent against tumor growth and viral infections, which abolishes the resistance of tumor cells to various forms of therapy.
  • Fields of application of the invention are medicine, the pharmaceutical and the biotechnical industry.
  • tumor cells can become resistant to apoptosis induced by chemotherapy, glucocorticoid therapy or radiation therapy.
  • the anti-apoptosis protein Bcl-2 (Sentman et al, Cell 67: 879, 1991; Mijashita et al, Blood 81: 151, 1993; Zhong et al, Proc. Natl. Acad. Sei USA) is primarily responsible for this effect 90: 4533, 1993).
  • the aim of the invention is to reduce or completely eliminate the resistance of tumor lines to different forms of therapy.
  • the object of the invention is to develop a corresponding agent against tumor growth and viral infections, which can possibly also be used for cleaning bone marrow preparations in transplantations.
  • this object is achieved with an agent according to the claims.
  • this agent contains a cell-permeable peptide which blocks anti-apoptosis proteins, in particular the protein Bcl-2.
  • Further anti-apoptosis proteins which can be blocked according to the invention are the proteins Bcl-X-., E1B, Mcl-1, AI, Ced-9 and BHRF1.
  • peptides which are homologous with Bcl-2 in the sequence regions 100-120 and 40-65 are particularly preferred. These peptides correspond to the amino acid sequences that allow interaction with nip proteins. They consist of a signal sequence that determines cell permeability and a functional sequence that corresponds to the two interaction domains from Bcl-2. The signal sequence has a length of 17 amino acids, the functional sequence has one
  • CORRECTED SHEET (RULE 91) ISA / EP Length of 9 or 10 amino acids.
  • Specific examples are the proteins FP1 with the sequence ERQIKIWFQNRRMKWKK-RRYRRDFAEM and FP2 with the sequence ERQIKIWFQNRRMKWKK-AAPAPGIFS.
  • the invention also relates to other peptides which comprise the functional sequences RRYRRDFAEM or AAPAPGIFS or parts thereof, e.g. B. a combination of these functional sequences with other signal sequences, which also cause cell permeability.
  • This also applies to peptides that only comprise parts of these functional sequences, preferably for peptides with 7 to 9 matching amino acids. But even if there is less agreement, the effectiveness of the product is usually still given; as a minimum, 3 amino acids must match the functional sequences.
  • the peptides according to the invention have extensive applications for improving the response rate of tumor cells to various forms of therapy. They are not only suitable for ex vivo / in vitro use (for example in purging), but also after stabilization for in vivo application. They are the first cell-compatible peptides that are suitable for modulating cell-compatible behavior by preventing protein-protein interactions. The findings are the first in vivo indication of the functional importance of certain regions of Bcl-2 for its anti-apoptosis effect.
  • the new cell-capable peptides can block many forms of protein-protein interaction that influence cellular behavior, for example in response to growth factors or caused by activated oncogenes.
  • Granulocyte macrophage stimulating factor prevents constitutive apoptosis of blood neutrophils (Brach et al., Blood 80: 2920, 1992). In the presence of the fusion peptides (FP 1, FP 2), but not in the presence of the control peptide (KP), GM-CSF is no longer able to prevent the constitutive apoptosis of blood neutrophils.
  • the addition of all peptides does not influence the spontaneous apoptosis of Bcl-2 negative blood neutrophils.
  • the dosage to be administered is not toxic per se and can therefore be implemented for an ex vivo / in vitro procedure (eg bone marrow spurging of myeloma patients).
  • Bcl-2 expressing tumor cells per se become dose-dependent depending on the dose, but not in the presence of signal peptide (SP), control peptide (KP) or functional peptide signal peptide (FP-SP) alone.
  • SP signal peptide
  • KP control peptide
  • FP-SP functional peptide signal peptide
  • the functionally relevant part of the peptide is not taken up by cells without preceding the signal sequence (functional peptide 1-SP) and has no effect intracellularly (no apoptosis of tumor cells expressing Bcl-2).
  • cell-capable peptides were developed that are suitable for modulating cellular behavior by preventing protein-protein interactions.
  • the findings are the first in vivo indication of the functional importance of certain regions of Bcl-21 for its anti-apoptosis effect.
  • the peptides developed allow tumor cells to be sensitized for chemotherapy.
  • the peptides have far-reaching implications for improving the response rate of tumor cells to therapy. They are suitable for ex vivo / in vitro use (for example in purging), but stabilization for in vivo application is also conceivable.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Biochemistry (AREA)
  • Biophysics (AREA)
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  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention pertains to a substance intended for use against tumour growth and viral infections. The substance in question eliminates the resistance of tumour cells to various forms of therapy. Suitable areas of application are medicine, pharmaceutic and biotechnological industry. The substance according to the invention contains as its active ingredient a cell-permeable peptide which blocks anti-apoptosis proteins. The peptide in question consists of a signal sequence of seventeen amino acids and a functional sequence of nine or ten amino acids. Specific peptides disclosed are ERQIKIWFQNRRMKWKK-RRYRRDFAEM(FP1) and ERQIKIWFQNRRMKWKK-AAPAPGIFS(FP2) which are homologous with Bcl-2. Also disclosed are peptides containing the functional sequences of FP1 or FP2 or parts thereof.

Description

Mittel gegen Tumorwachstum und virale InfektionenAnti-tumor and viral infection remedies
Beschreibungdescription
Die Erfindung betrifft ein Mittel gegen Tumorwachstum und virale Infektionen, welches die Resistenz von Tumorzellen gegenüber verschiedenen Therapieformen aufhebt. Anwendungs¬ gebiete der Erfindung sind die Medizin, die pharmazeutische und die biotechnische Industrie.The invention relates to an agent against tumor growth and viral infections, which abolishes the resistance of tumor cells to various forms of therapy. Fields of application of the invention are medicine, the pharmaceutical and the biotechnical industry.
Eε ist bekannt, daß Tumorzellen gegen die durch Chemo-, Glukokortikoid- oder Strahlentherapie induzierte Apoptose resistent werden können. Maßgeblich verantwortlich für diesen Effekt ist das Anti-Apoptose-Protein Bcl-2 (Sentman et al, Cell 67: 879,1991; Mijashita et al, Blood 81: 151, 1993; Zhong et al, Proc. Natl. Acad. Sei USA 90: 4533, 1993) .It is known that tumor cells can become resistant to apoptosis induced by chemotherapy, glucocorticoid therapy or radiation therapy. The anti-apoptosis protein Bcl-2 (Sentman et al, Cell 67: 879, 1991; Mijashita et al, Blood 81: 151, 1993; Zhong et al, Proc. Natl. Acad. Sei USA) is primarily responsible for this effect 90: 4533, 1993).
Man hat versucht, Bcl-2-vermittelte Resistenz gegenüber Chemotherapie-induzierter Apoptose durch spezifische Unterdrückung der Translation des Bcl-2 Transkriptes in das Bcl-2 Protein mittelε Antisenεe-Oligonukleotiden aufzuheben. Auf Grund der langen Halbwertszeit von Bcl-2 ist dies jedoch nur sehr unzufriedenstellend , mit geringer Spezifitat möglich und erfordert eine lange Exposition der Tumorzellen mit den Antisense-Oligonukleotiden. (Reed et al, Canc.Res 50: 6565, 1995; Campos et al, Blood 84: 595, 1994). Damit eignet sich diese Verfahrensweise nicht für eine Applikation am Patienten, etwa für die Knochenmarksaufreinigung, die nur eine kurzfristige ex vivo Kultivierung der zur Transplantation vorgesehenen Zellen (bis zu 48 Stunden) erlaubt.Attempts have been made to abolish Bcl-2-mediated resistance to chemotherapy-induced apoptosis by specifically suppressing the translation of the Bcl-2 transcript into the Bcl-2 protein by means of antiseic oligonucleotides. However, due to the long half-life of Bcl-2, this is only very unsatisfactory, possible with little specificity and requires long exposure of the tumor cells to the antisense oligonucleotides. (Reed et al, Canc.Res 50: 6565, 1995; Campos et al, Blood 84: 595, 1994). This procedure is therefore not suitable for application to the patient, for example for bone marrow purification, which only allows short-term ex vivo cultivation of the cells intended for transplantation (up to 48 hours).
Es wurden kürzlich zwei Regionen sowohl in Bcl-2 als auch in einem anderen Protein, E1B, welcheε ebenfalls Apoptoεe verhindert, identifiziert (Boyd et al, Cell 79: 341, 1994; Tarodi et al, Virology 201: 404, 1994). Diese Regionen sind homolog, und sie sind in der Lage, Protein-Protein Interaktionen von Bcl-2 bzw. E1B mit putativen Effektor- Molekülen, Nip 1,2 und 3, BAX und BIK genannt, zu vermitteln (gezeigt in vitro im Yeast-Two Hybrid-System) . Mutationen innerhalb dieser Domänen im E1B Protein, die die Interaktion von E1B mit Nip aufheben, verhindern auch die anti- apoptotische Wirkung von E1B in zellulären Systemen (Subdramarian et al, Gene 124: 173, 1993). Die funktionelle Bedeutung dieser Regionen in Bcl-2 Protein ist dagegen nicht bekannt. Zudem ist unklar, ob diese Regionen notwendig und ausreichend für die anti-apoptotische Wirkung beider Proteine sind.Two regions have recently been identified both in Bcl-2 and in another protein, E1B, which also prevents apoptosis (Boyd et al, Cell 79: 341, 1994; Tarodi et al, Virology 201: 404, 1994). These regions are homologous and they are able to mediate protein-protein interactions of Bcl-2 or E1B with putative effector molecules, called Nip 1,2 and 3, BAX and BIK (shown in vitro in the Yeast- Two hybrid system). Mutations within these domains in the E1B protein that cancel the interaction of E1B with Nip also prevent the anti-apoptotic effect of E1B in cellular systems (Subdramarian et al, Gene 124: 173, 1993). However, the functional importance of these regions in Bcl-2 protein is not known. It is also unclear whether these regions are necessary and sufficient for the anti-apoptotic effects of both proteins.
Das Ziel der Erfindung besteht darin, die Resistenz von Tumorzeilen gegenüber verschiedenen Therapieformen zu vermindern bzw. ganz aufzuheben. Aufgabe der Erfindung ist die Entwicklung eines entsprechenden Mittels gegen Tumorwachstum und virale Infektionen, das möglicherweise auch zur Reinigung von Knochenmarkspräparationen bei Trans¬ plantationen eingesetzt werden kann.The aim of the invention is to reduce or completely eliminate the resistance of tumor lines to different forms of therapy. The object of the invention is to develop a corresponding agent against tumor growth and viral infections, which can possibly also be used for cleaning bone marrow preparations in transplantations.
Erfindungsgemäß wird diese Aufgabe mit einem Mittel gemäß den Ansprüchen gelöst. Dieses Mittel enthält als Wirkstoff ein zellpermeables Peptid, das Anti-Apoptose-Proteine, insbesondere das Protein Bcl-2, blockiert. Weitere Anti- Apoptose-Proteine, die gemäß der Erfindung blockiert werden können, sind die Proteine Bcl-X-., E1B, Mcl-1, AI, Ced-9 und BHRF1.According to the invention, this object is achieved with an agent according to the claims. As active ingredient, this agent contains a cell-permeable peptide which blocks anti-apoptosis proteins, in particular the protein Bcl-2. Further anti-apoptosis proteins which can be blocked according to the invention are the proteins Bcl-X-., E1B, Mcl-1, AI, Ced-9 and BHRF1.
Besonders bevorzugt ist der Einsatz von Peptiden, die mit Bcl-2 in den Sequenzbereichen 100-120 und 40-65 homolog sind. Diese Peptide entsprechen den Aminosäuresequenzen, die die Interaktion mit Nip-Proteinen erlauben. Sie bestehen aus einer die Zellpermeabilität bedingenden Signalsequenz und einer den zwei Interaktions-Domänen aus Bcl-2 entsprechenden Funktionssequenz. Die Signalsequenz hat eine Länge von 17 Aminosäuren, die Funktionssequenz hat eineThe use of peptides which are homologous with Bcl-2 in the sequence regions 100-120 and 40-65 is particularly preferred. These peptides correspond to the amino acid sequences that allow interaction with nip proteins. They consist of a signal sequence that determines cell permeability and a functional sequence that corresponds to the two interaction domains from Bcl-2. The signal sequence has a length of 17 amino acids, the functional sequence has one
BERICHTIGTES BLATT (REGEL 91) ISA/EP Länge von 9 bzw. 10 Aminosäuren. Konkrete Beispiele sind die Proteine FPl mit der Sequenz ERQIKIWFQNRRMKWKK-RRYRRDFAEM und FP2 mit der Sequenz ERQIKIWFQNRRMKWKK-AAPAPGIFS. Gegenstand der Erfindung sind auch andere Peptide, die die Funktionssequenzen RRYRRDFAEM oder AAPAPGIFS bzw. Teile davon umfassen, z. B. eine Kombination dieser Funktionssequenzen mit anderen Signalsequenzen, die ebenfalls eine Zellpermeabilität bedingen. Das gilt auch für Peptide, die nur Teile dieser Funktionssequenzen umfassen, bevorzugt für Peptide mit 7 bis 9 übereinstimmenden Aminosäuren. Aber auch bei einer geringeren Obereinstimmung ist im Regelfalle noch eine Wirksamkeit des Mittels gegeben; als Minimum müssen 3 Aminosäuren mit den Funktionssequenzen übereinstimmen.CORRECTED SHEET (RULE 91) ISA / EP Length of 9 or 10 amino acids. Specific examples are the proteins FP1 with the sequence ERQIKIWFQNRRMKWKK-RRYRRDFAEM and FP2 with the sequence ERQIKIWFQNRRMKWKK-AAPAPGIFS. The invention also relates to other peptides which comprise the functional sequences RRYRRDFAEM or AAPAPGIFS or parts thereof, e.g. B. a combination of these functional sequences with other signal sequences, which also cause cell permeability. This also applies to peptides that only comprise parts of these functional sequences, preferably for peptides with 7 to 9 matching amino acids. But even if there is less agreement, the effectiveness of the product is usually still given; as a minimum, 3 amino acids must match the functional sequences.
Die erfindungsgemäßen Peptide haben weitreichende Anwendungsmöglichkeiten zur Verbesserung der Ansprechrate von Tumorzellen auf verschiedene Therapieformen. Sie eignen sich nicht nur für einen ex vivo/in vitro-Einsatz (beispielsweise beim Purging), sondern auch nach Stabilisierung für eine in vivo Applikation. Eε εind die ersten zellgängigen Peptide, die geeignet sind, zellgängigeε Verhalten zu modulieren, indem sie Protein- Protein-Interaktionen unterbinden. Die Befunde sind der erste in vivo Hinweis auf die funktionelle Bedeutung bestimmter Regionen von Bcl-2 für seine Anti-Apoptose- Wirkung.The peptides according to the invention have extensive applications for improving the response rate of tumor cells to various forms of therapy. They are not only suitable for ex vivo / in vitro use (for example in purging), but also after stabilization for in vivo application. They are the first cell-compatible peptides that are suitable for modulating cell-compatible behavior by preventing protein-protein interactions. The findings are the first in vivo indication of the functional importance of certain regions of Bcl-2 for its anti-apoptosis effect.
Das hier erstmals aufgezeigte Prinzip läßt sich auf eine Vielzahl anderer Anwendungsmöglichkeiten übertragen. So lassen sich mit den neuen zellgängigen Peptiden prinzipiell viele Formen der Protein-Protein-Interaktion blockieren, die zelluläres Verhalten, etwa in Antwort auf Wachstumsfaktoren oder verursacht durch aktivierte Onkogene, beeinflussen.The principle shown here for the first time can be applied to a variety of other possible applications. In principle, the new cell-capable peptides can block many forms of protein-protein interaction that influence cellular behavior, for example in response to growth factors or caused by activated oncogenes.
Die Erfindung soll nachfolgend durch Ausführungsbeispiele und Abbildungen näher erläutert werden.The invention is intended to be described in the following by means of exemplary embodiments and illustrations are explained in more detail.
Ausführungsbeispieleembodiments
Es wurden die erfindungsgemäßen Peptide FPl und FP2 verglichenThe peptides FP1 and FP2 according to the invention were compared
- mit Peptiden, die nur die Signalsequenz enthalten (SP)- with peptides that only contain the signal sequence (SP)
- mit Peptiden, bei denen die Signalsequenz an solche Aminosäuren aus dem Bcl-2 Protein fusioniert wurde, die für die Interaktion mit Nip-Proteinen nicht notwendig sind (Kontrollpeptid KP) undwith peptides in which the signal sequence has been fused to those amino acids from the Bcl-2 protein which are not necessary for the interaction with nip proteins (control peptide KP) and
- mit Peptiden, die die Bcl-2 homologe Domäne des Funktionspeptids 1 repräsentieren, nicht aber an das Signalpeptid gekoppelt sind(FP-SP).- With peptides that represent the Bcl-2 homologous domain of the functional peptide 1, but are not coupled to the signal peptide (FP-SP).
Abbildung 1illustration 1
Die Zugabe aller Peptide ist dosisabhängig (bis zu lOOμg/ml) per se nicht toxisch in Zellen, die kein Bcl-2 exprimieren (humane Bcl-2 negative Tumorzell-Linie Karpaε) .The addition of all peptides is dose-dependent (up to 100 μg / ml) per se not toxic in cells that do not express Bcl-2 (human Bcl-2 negative tumor cell line Karpaε).
Abbildung 2AFigure 2A
Granulozyten-Makrophagen stimulierender Faktor verhindert die konstitutive Apoptose von Blut-Neutrophilen (Brach et al., Blood 80: 2920, 1992). In Anwesenheit der Fusionspeptide (FP 1, FP 2), nicht aber in Anwesenheit des Kontrollpeptides (KP), ist GM-CSF nicht mehr in der Lage, die konstitutive Apoptose von Blut-Neutrophilen zu verhindern.Granulocyte macrophage stimulating factor prevents constitutive apoptosis of blood neutrophils (Brach et al., Blood 80: 2920, 1992). In the presence of the fusion peptides (FP 1, FP 2), but not in the presence of the control peptide (KP), GM-CSF is no longer able to prevent the constitutive apoptosis of blood neutrophils.
Abbildung 2BFigure 2B
Die Zugabe aller Peptide beeinflußt nicht den spontanen Apoptoεe-Verlauf von Bcl-2 negativen Blut-Neutrophilen. Die zu verabreichende Dosierung ist per se nicht toxisch und somit für eine ex vivo/in vitro Vorgehensweise (z.B. Knochenmarkspurging von Myelom-Patienten) realisierbar.The addition of all peptides does not influence the spontaneous apoptosis of Bcl-2 negative blood neutrophils. The The dosage to be administered is not toxic per se and can therefore be implemented for an ex vivo / in vitro procedure (eg bone marrow spurging of myeloma patients).
Abbildung 3Figure 3
In Anwesenheit der Fusionspeptide (FP 1, FP 2) werden Bcl-2 exprimierende Tumorzellen per se dosisabhängig apoptisch, nicht jedoch in Anwesenheit von Signalpeptid (SP), Kontrollpeptid (KP) oder Funktionspeptid-Signalpeptid (FP- SP) allein. Der funktionell relevante Teil des Peptides wird ohne Vorschaltung der Signalsequenz (Funktionspeptid 1-SP) nicht von Zellen aufgenommen und entfaltet intrazellulär keine Wirkung (keine Apoptose von Bcl-2 exprimierenden Tumorzellen) .In the presence of the fusion peptides (FP 1, FP 2), Bcl-2 expressing tumor cells per se become dose-dependent depending on the dose, but not in the presence of signal peptide (SP), control peptide (KP) or functional peptide signal peptide (FP-SP) alone. The functionally relevant part of the peptide is not taken up by cells without preceding the signal sequence (functional peptide 1-SP) and has no effect intracellularly (no apoptosis of tumor cells expressing Bcl-2).
Abbildung 4Figure 4
In Anwesenheit des Funktionspeptides (FP 1) erfolgt zusammen mit Dexamethason synergistische Induktion von Apoptose in Tumor2ellen. In Anwesenheit der Kontrollpeptide (Signalpeptid oder Kontrollpeptid) sind diese Tumorzellen resistent gegenüber Dexamethason induzierter Apoptose.In the presence of the functional peptide (FP 1), synergistic induction of apoptosis in tumor cells takes place together with dexamethasone. In the presence of the control peptides (signal peptide or control peptide), these tumor cells are resistant to dexamethasone-induced apoptosis.
Abbildung 5Figure 5
In dieser Abbildung ist dargestellt, daß die Resistenz von E1B überexprimierenden 293-Zellen gegenüber Tumor-Nekrose Faktor induzierter Apoptose durch Einsatz zellpermeabler Peptide überkommen wird.This figure shows that the resistance of E1B overexpressing 293 cells to tumor necrosis factor-induced apoptosis will be overcome by using cell-permeable peptides.
BERICHTIGTES BLATT (REGEL 91) ISA/EP Der funktionell relevante Teil deε Peptides wird ohne Vorschaltung der Signalsequenz nicht von Zellen aufgenommen und entfaltet intrazellulär keine Wirkung (keine Apoptose von Bcl-2 exprimierenden Tumorzellen) .CORRECTED SHEET (RULE 91) ISA / EP The functionally relevant part of the peptide is not taken up by cells without preceding the signal sequence and has no effect intracellularly (no apoptosis of tumor cells expressing Bcl-2).
Befunde in allgemeiner Form:Findings in general form:
Erstmals wurden hier zellgängige Peptide entwickelt, die geeignet sind, zelluläres Verhalten zu modulieren, indem sie Protein-Protein-Interaktionen unterbinden. Die Befunde sind der erste in vivo Hinweis auf die funktionelle Bedeutung bestimmter Regionen von Bcl-21 für seine Anti-Apoptose- Wirkung. Die entwickelten Peptide erlauben eine Sensitivierung von Tumorzellen für Chemotherapie.For the first time, cell-capable peptides were developed that are suitable for modulating cellular behavior by preventing protein-protein interactions. The findings are the first in vivo indication of the functional importance of certain regions of Bcl-21 for its anti-apoptosis effect. The peptides developed allow tumor cells to be sensitized for chemotherapy.
AuswirkungenImpact
Die Peptide haben weitreichende Implikation zur Verbesserung der Ansprechrate von Tumorzellen auf Therapie. Sie eignen sich für einen ex vivo/in vitro-Einsatz (beispielsweise beim Purging) , denkbar ist aber auch eine Stabilisierung für eine in vivo Applikation.The peptides have far-reaching implications for improving the response rate of tumor cells to therapy. They are suitable for ex vivo / in vitro use (for example in purging), but stabilization for in vivo application is also conceivable.
Das hier erstmals aufgezeigte Prinzip läßt sich auf eine Vielzahl anderer Anwendungsmöglichkeiten übertragen. So lassen sich mit zellgängigen Peptiden prinzipiell viele Formen der Protein-Protein-Interaktion blockieren, die zelluläres Verhalten, etwa in Antwort auf Wachstumsfaktoren oder aber verursacht durch aktivierte Onkogene, beein¬ flussen.The principle shown here for the first time can be applied to a variety of other possible applications. In principle, many forms of protein-protein interaction can be blocked with cell-permeable peptides that influence cellular behavior, for example in response to growth factors or caused by activated oncogenes.
BERICHTIGTES BLATT (REGEL 91) ISA/EP CORRECTED SHEET (RULE 91) ISA / EP

Claims

Patentansprüche claims
1. Mittel gegen Tumorwachstum und virale Infektionen, enthaltend als Wirkstoff ein zellpermeableε Peptid, daε Anti-Apoptose-Proteine blockiert.1. Agent against tumor growth and viral infections, containing as active ingredient a cell-permeable peptide that blocks anti-apoptosis proteins.
2. Mittel nach Anspruch 1, enthaltend als Wirkstoff ein zellpermeableε Peptid, daε daε Anti-Apoptose-Protein Bcl-2 blockiert.2. Composition according to claim 1, containing as active ingredient a cell-permeable peptide that blocks anti-apoptosis protein Bcl-2.
3. Mittel nach Anspruch 1, enthaltend alε Wirkεtoff ein Peptid, daε mit Bcl-2 in den Sequenzbereichen 100-120 und 40-65 homologe Anti-Apoptose-Proteine blockiert.3. Composition according to claim 1, containing as an active substance a peptide that blocks homologous anti-apoptosis proteins with Bcl-2 in the sequence regions 100-120 and 40-65.
4. Mittel nach Anspruch 1 bis 3 , dadurch gekennzeichnet, daß daε Peptid auε einer die Zellpermeabilität bedingenden Signalεequenz und einer den Bcl-2-Domänen entsprechenden Funktionssequenz besteht.4. Means according to claims 1 to 3, characterized in that the peptide consists of a signal sequence which determines the cell permeability and a functional sequence which corresponds to the Bcl-2 domains.
5. Mittel nach Anspruch 1 bis 4, dadurch gekennzeichnet, daß das Peptid auε einer 17 Aminoεäuren großen Signalsequenz und einer 9 bzw. 10 Aminosäuren großen Funktionεεequenz beεteht.5. Composition according to claim 1 to 4, characterized in that the peptide consists of a 17 amino acid large signal sequence and a 9 or 10 amino acid large functional sequence.
6. Mittel nach Anspruch 1 bis 5, dadurch gekennzeichnet, daß eε folgende Sequenz enthält: ERQIKIWFQNRRMKWKK- RRYRRDFAEM(FPl) .6. Composition according to claim 1 to 5, characterized in that eε contains the following sequence: ERQIKIWFQNRRMKWKK- RRYRRDFAEM (FPl).
7. Mittel nach Anspruch 1 bis 6, dadurch gekennzeichnet, daß eε folgende Sequenz enthält: ERQIKIWFQNRRMKWKK- AAPAPGIFS(FP2) .7. Composition according to claim 1 to 6, characterized in that eε contains the following sequence: ERQIKIWFQNRRMKWKK- AAPAPGIFS (FP2).
8. Mittel nach Anεpruch 1 bis 4, dadurch gekennzeichnet, daß es ein Peptid enthält, das die Funktionssequenzen RRYRRDFAEM oder8. Composition according to Anεpruch 1 to 4, characterized in that it contains a peptide that the functional sequences RRYRRDFAEM or
AAPAPGIFS bzw. Teile davon umfaßt. AAPAPGIFS or parts thereof.
9. Zellpermeableε Peptid mit der Eigenschaft, daß es Anti- Apoptose-Proteine blockiert.9. cell permeable peptide with the property that it blocks anti-apoptosis proteins.
10. Zellpermeables Peptid nach Anspruch 9, dadurch gekennzeichnet, daß es das Anti-Apoptose-Protein Bcl-2 blockiert.10. Cell-permeable peptide according to claim 9, characterized in that it blocks the anti-apoptosis protein Bcl-2.
11. Zellpermeables Peptid nach Anspruch 9 und 10, dadurch gekennzeichnet, daß es mit Bcl-2 in den Sequenzbereichen 100-120 und 40-65 homologe Anti-Apoptose-Proteine blockiert.11. Cell-permeable peptide according to claim 9 and 10, characterized in that it blocks homologous anti-apoptosis proteins with Bcl-2 in the sequence regions 100-120 and 40-65.
12. Zellpermeables Peptid nach Anspruch 9 bis 11, dadurch gekennzeichnet, daß eε aus einer die Zellpermeabilität bedingenden Signalsequenz und einer den Bcl-2-Domänen entsprechenden Funktionsεequenz beεteht.12. Cell-permeable peptide according to claim 9 to 11, characterized in that eε consists of a signal sequence which determines the cell permeability and a function sequence corresponding to the Bcl-2 domains.
13. Zellpermeableε Peptid nach Anεpruch 9 bis 12, dadurch gekennzeichnet, daß eε auε einer 17 Aminosäuren großen Signalsequenz und einer 9 bzw. 10 Aminosäuren großen Funktionssequenz besteht.13. cell permeable peptide according to claim 9 to 12, characterized in that eε consists of a 17 amino acid signal sequence and a 9 or 10 amino acid functional sequence.
14. Zellpermeables Peptid nach Anspruch 9 biε 13, gekennzeich-net durch folgende Sequenz: ERQIKIWFQNRRMKWKK- RRYRRDFAEM(FPl) .14. Cell-permeable peptide according to claim 9 to 13, characterized by the following sequence: ERQIKIWFQNRRMKWKK-RRYRRDFAEM (FPI).
15. Zellpermeableε Peptid nach Anspruch 9 bis 13, gekennzeich-net durch folgende Sequenz ERQIKIWFQNRRMKWKK- AAPAPGIFS(FP2).15. Zellpermeableε peptide according to claim 9 to 13, marked by the following sequence ERQIKIWFQNRRMKWKK- AAPAPGIFS (FP2).
16. Zellpermeables Peptid nach Anεpruch 9 biε 13, gekennzeichnet durch die Funktionεsequenzen RRYRRDFAEM oder AAPAPGIFS bzw. Teile davon. 16. Cell-permeable peptide according to claim 9 to 13, characterized by the functional sequences RRYRRDFAEM or AAPAPGIFS or parts thereof.
PCT/DE1996/001286 1995-07-18 1996-07-16 Substance for use against tumour growth and viral infections WO1997004006A1 (en)

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EP0902088A2 (en) * 1996-03-26 1999-03-17 Razvan T. Radulescu Peptides with antiproliferative properties
WO1999064449A2 (en) * 1998-06-10 1999-12-16 The Queen's University Of Belfast Cell-permeable peptide

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JP2005529616A (en) * 2002-06-18 2005-10-06 アイアールエム エルエルシー Diagnosis and treatment of chemotherapy-resistant tumors

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997035873A2 (en) * 1996-03-26 1997-10-02 Radulescu Razvan T Peptides with antiproliferative properties
WO1997035873A3 (en) * 1996-03-26 1997-12-31 Razvan T Radulescu Peptides with antiproliferative properties
EP0902088A2 (en) * 1996-03-26 1999-03-17 Razvan T. Radulescu Peptides with antiproliferative properties
EP0902088A3 (en) * 1996-03-26 1999-09-15 Razvan T. Radulescu Peptides with antiproliferative properties
WO1999064449A2 (en) * 1998-06-10 1999-12-16 The Queen's University Of Belfast Cell-permeable peptide
WO1999064449A3 (en) * 1998-06-10 2002-10-24 Univ Belfast Cell-permeable peptide

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