WO1998042320A2 - Prolongation de l'expression de proteines transgeniques par traitement immunomodulant avec de la 15-desoxyspergualine - Google Patents

Prolongation de l'expression de proteines transgeniques par traitement immunomodulant avec de la 15-desoxyspergualine Download PDF

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Publication number
WO1998042320A2
WO1998042320A2 PCT/EP1998/001439 EP9801439W WO9842320A2 WO 1998042320 A2 WO1998042320 A2 WO 1998042320A2 EP 9801439 W EP9801439 W EP 9801439W WO 9842320 A2 WO9842320 A2 WO 9842320A2
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WIPO (PCT)
Prior art keywords
use according
cells
medicament
transgenic
combination
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PCT/EP1998/001439
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German (de)
English (en)
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WO1998042320A3 (fr
Inventor
Gerhard Seemann
Günter CICHON
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Hoechst Aktiengesellschaft
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Application filed by Hoechst Aktiengesellschaft filed Critical Hoechst Aktiengesellschaft
Priority to AU70344/98A priority Critical patent/AU7034498A/en
Publication of WO1998042320A2 publication Critical patent/WO1998042320A2/fr
Publication of WO1998042320A3 publication Critical patent/WO1998042320A3/fr

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    • 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

Definitions

  • the invention relates to the use of 15-deoxyspergualin for the manufacture of a medicament in order to increase the tolerance of a mammal to transgenic cells.
  • the use of 15-deoxyspergualin significantly extends the production of the transgenic expression product even after stopping the immunosuppressive treatment.
  • transgenic cells are killed, for example, by cytotoxic immune cells, and thus the cellular immune reaction that terminates the expression of one or more proteins or peptides caused by the introduced genetic material.
  • a humoral immune response in turn makes it more difficult or prevents the reintroduction of the genetic material, e.g. B. by neutralizing the vectors using specific antibodies (Tripathy SK et al., Nat. Med: 2 (5), 1996, pages 545-550; Yang Y. et al., Gene Ther. 3 (2): 1996, Pages 137-144).
  • WO 96/12406 it was described that the humoral immune response against an adenovirus vector can be suppressed by an immunosuppressive accompanying therapy and that this enables repeated vector administration becomes.
  • steroids such as dexamethasone, especially cyclosporin A and 1-amino-19-guanidine-11-hydroxy-4, 9,12-triazanone decane-10,13-dione-trihydrochloride (15-deoxyspergualin; hereinafter referred to as DSG) recommended for this purpose.
  • the object of the invention is the identification of substances which sustainably prevent the rapid destruction of the transgenic cells and thus increase the tolerance of a mammal to transgenic cells, even after stopping the immunosuppressive accompanying therapy.
  • the expression of the transgenic product or products is thus retained longer in vivo. As a result, repeated application of the genetic material could be avoided or at least limited in frequency.
  • DSG has been found to have the desired effectiveness.
  • FK 506 and cyclosporin A which is particularly recommended in WO 96/12406, are not suitable.
  • the object is therefore achieved according to the invention by the use of the DSG for the production of a medicament or a medicament combination for increasing the tolerance of a mammal, in particular human beings, to transgenic cells.
  • DSG also means physiologically compatible salts, DSG derivatives, isomers and metabolites.
  • transgenic cells are able to produce the transgenic expression products in vivo longer than they are able to do in mammals of a non-immunosuppressed control group.
  • Transgenic cells are cells of any kind, that is to say animal, plant or bacterial cells, preferably cells of mammals, very particularly preferred human cells into which genetic material has been introduced.
  • the aim associated with the production of the transgenic cells is irrelevant to the general inventive concept. Furthermore, neither the production process of the transgenic cells nor the genetic material per se is of crucial importance for the invention.
  • the introduced genetic material can consist, for example, of one or more deoxyribonucleic acid and / or ribonucleic acid chains.
  • the nucleic acid chains can contain promoters, various other gene regulatory control functions, sequences for specific incorporation into the cellular genome and / or gene sequences which code for specific proteins or peptides.
  • the introduced genetic material can be alien or identical to the cellular genome or can even originate from the same individual. A combination of these is also possible.
  • the specific transgenic expression product (s) can be non-natural or natural, for example human, animal, vegetable, bacterial or viral proteins or peptides.
  • examples are enzymes, receptors, messenger substances, hormones, growth factors, coagulation factors, apolipoproteins, factors influencing the metabolism or cell division, anti-inflammatory factors, inhibitors or activators of the cell cycle as well as intracellular signal chains, tumor suppressors, elements of the cytoskeleton or of the connective tissue regulator, or antigens of disease agents, or antigens of antigen as well as tumor-associated antigens.
  • insulin examples are insulin; Hirudin; Factor VIII; Factor IX; Factor XIII; von Willebrand factor; Antibody; Erythropoitin; human growth hormone; "Growth factors” such as EGF, TGF ⁇ , TGFß, GM-CSF, PDGF, nerve growth factor and others; "Tumor necrosis factor”; Interferons; Interleukins; p53; Tumor therapy, hepatitis virus antigens; HIV antigens, herpes virus antigens; Borrelia antigens; Plasmodium antigens; Trypanosome antigens, Taenia antigens or human ⁇ -glucuronidase
  • the invention is further characterized that about 15 days after stopping the immunosuppressive drug or drug combination in the treated mammals more than 1.5 times, preferably more than twice, most preferably more than ⁇ times the transgenic expression product can be generated as in mammals in a non-immunosuppressed control group.
  • the detection of the expressed transgenic product does not have to be done on the 15th day; it is also possible to take measurements before or after this time. It is crucial that not just a few days, for example 10 or 20 days, after stopping the immunosuppressive treatment, the transgenic expression product is only produced in the same or a smaller amount as in non-immunosuppressed organisms of a control group.
  • the use according to the invention further comprises administration of the drug or the drug combination before, during and / or after the application of the transgenic cells produced in vitro or their generation in vivo. Also included is the use according to the invention for the support of a gene therapy treatment producing transgenic cells.
  • in vitro or in vivo genetic material in the form of one or more nucleic acid chains is introduced into cells. This takes place, for example, with the aid of viral vectors, such as adenoviruses, retroviruses or herpes viruses, or other methods, for example via transfection, by direct injection, by "gene gun", or with the aid of liposomes, virosomes or receptor-mediated transport systems.
  • viral vectors such as adenoviruses, retroviruses or herpes viruses
  • the gene therapy to be supported can treat diseases in which a protein or peptide is not, insufficiently or only faultily produced in the body of the mammal, but it is also used to vaccinate against various pathogens or against degenerate cells in the body and for the production of non-natural proteins or peptides in body cells that promote or inhibit the activities of enzyme or signal cascades or for the production of enzymes that specific Activate prodrugs.
  • the gene therapy to be supported can be used to treat congenital diseases such as cystic fibrosis, nerve and brain diseases such as Parkinson's, Alzheimer's or Jakop-Kreuzfeld syndrome; rheumatic diseases, osteoathritis, osteopoiesis or arthrosis, familial hypercholesterolemia, hemophilia, sickle cell anemia, phenylketonuria; metabolic disorders like diabetes; of inflammation; of cancers; infectious diseases, such as AIDS or hepatitis, or hormone and growth disorders.
  • Another area of gene therapy to be supported is the generation of vaccination protection against pathogens such as viruses, bacteria, fungi, single and multicellular parasites and against degenerate endogenous cells such as tumor cells.
  • the invention further includes a process for the manufacture of a medicament or combination of medicaments to increase the tolerance of a mammal to transgenic cells, which is characterized in that DSG is used alone or with other immunosuppressants with a physiologically acceptable carrier and other suitable active ingredient - or adjuvants in a suitable dosage form.
  • DSG or its physiologically compatible and pharmacologically active salts or derivatives are administered, for example, in a dose of 0.1 to 100 mg / kg, preferably 0.1 to 35 mg / kg, very particularly preferably 0.1 to 10 mg / kg.
  • the medicament according to the invention or the medicament combination according to the invention can be administered, for example, orally, intravenously, subcutaneously, intraperitoneally, percutaneously, cutaneously, topically, by inhalation, intramuscularly, intrathecally, intraocularly, ocularly, buccally, nasally or rectally, preferably intravenously or orally.
  • Example 1 serve to illustrate the invention. They are not to be understood as a limitation of any kind.
  • Example 1 serve to illustrate the invention. They are not to be understood as a limitation of any kind.
  • Cyclosporin A 50-100 mg / kg / day
  • DSG 10 mg / kg / day
  • adenovirus reporter gene ⁇ -galactosidase
  • the control group reached a maximum of the reporter gene expression in the liver on the 6th day, after which the expression was continuously reduced by the activity of cytotoxic T-cells and reached the initial level after about 21 days.
  • reporter gene expression in the cyclosporin A group fell to baseline over a period of 21 days (42 days after vector administration), while massive gene expression was still detectable in the DSG group.
  • a dose of 1 x 10 10 adenoviruses was administered to all mice via the tail vein.
  • the recombinant adenoviruses carry the gene for human alpha 1 -antitrypsin as a serum reporter gene. This gene has proven to be a viable reporter because it does not act as an antigen in the mouse, but it does can be distinguished from the marine alpha 1 -antitrypsin with a specific antibody.
  • the half-life of the antiprotease is about 3-4 days, so that the serum level reflects the current synthesis performance of the liver quite well.
  • the total collective was divided into five groups, each with 4 animals, which were treated as follows:
  • 1st group control group - no immunosuppressive treatment, application of saline i. p. for a period of 5 days after virus administration.
  • 2nd group FK 506 group - application of 1 mg / kg day FK 506 i. p. starting 1 day before virus administration and then for a period of 5 days.
  • 3rd group Cyclosporin A group - application of 20 mg / kg / day i. p. starting 1 day before virus administration and then for a period of 5 days.
  • 4th group DSG group - application of 10 mg / kg / day i. p. starting 1 day before virus administration and then for 5 days.
  • 5th group DSG ++ group - application of 10 mg / kg / day i. p. starting 1 day before virus administration and then daily for five days and then twice a week for 150 days.
  • NMRI mice were used because, due to their genetic heterogeneity, these mice develop an immunological reaction pattern that is closer to that of a human population than pure inbred strains, the price is an increased variability in immunological responsiveness. Nevertheless, the individual groups develop significant reaction patterns.
  • Table 1 shows the mean values of the serum level of human ⁇ 1 -antitrypsin.
  • a 5-day treatment with DSG leads to a significant increase and prolongation of the antitrypsin expression.
  • the quantity factor is approximately 1.8 days after stopping DSG.
  • Continuous treatment (twice a week) can no longer dramatically increase this effect.
  • the continuous DSG treatment increases the expression only slightly via transient administration, but the antibody load against viral proteins is reduced by approximately 90% after 150 days (see Table 2). Even the transient administration of DSG reduces the antibody response by more than 60% compared to the control with a clearly positive effect on expression.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne l'utilisation de 15-désoxyspergualine pour préparer un médicament destiné à augmenter la tolérance d'un mammifère vis-à-vis de cellules transgéniques. L'utilisation de 15-désoxyspergualine permet de prolonger de manière notoire la production du produit d'expression transgénique, même après arrêt du traitement immunosuppresseur.
PCT/EP1998/001439 1997-03-21 1998-03-12 Prolongation de l'expression de proteines transgeniques par traitement immunomodulant avec de la 15-desoxyspergualine WO1998042320A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU70344/98A AU7034498A (en) 1997-03-21 1998-03-12 Extension of the expression of transgenic proteins by immunomodulation with 15-deoxyspergualine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19711803.8 1997-03-21
DE1997111803 DE19711803A1 (de) 1997-03-21 1997-03-21 Verlängerung der Expression von transgenen Proteinen durch immunmodulierende Behandlung mit 15-Deoxyspergualin

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WO1998042320A2 true WO1998042320A2 (fr) 1998-10-01
WO1998042320A3 WO1998042320A3 (fr) 1998-12-23

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DE (1) DE19711803A1 (fr)
WO (1) WO1998042320A2 (fr)

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* Cited by examiner, † Cited by third party
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DE19923961A1 (de) 1999-05-25 2000-11-30 Euro Nippon Kayaku Gmbh Verwendung von 15-Deoxyspergualin zur Behandlung von hyperreaktiven entzündlichen Erkrankungen und Autoimmunerkrankungen

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996012406A1 (fr) * 1994-10-19 1996-05-02 Genetic Therapy, Inc. Therapie genique par administration concurrente et repetee d'adenovirus et d'agents immunodepresseurs
WO1997039776A1 (fr) * 1996-04-19 1997-10-30 Genetic Therapy, Inc. Therapie genique reposant sur l'administration conjointe et repetee d'adenovirus et d'agents immunosuppresseurs

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996012406A1 (fr) * 1994-10-19 1996-05-02 Genetic Therapy, Inc. Therapie genique par administration concurrente et repetee d'adenovirus et d'agents immunodepresseurs
WO1997039776A1 (fr) * 1996-04-19 1997-10-30 Genetic Therapy, Inc. Therapie genique reposant sur l'administration conjointe et repetee d'adenovirus et d'agents immunosuppresseurs

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
HALLORAN, P.F.: "Rethinking Immunosuppression in Terms of the Redundant and Nonredundant Steps in the Immune Response" TRANSPLANTATION PROCEEDINGS, Bd. 28, Nr. 6, 1996, Seiten 11-18, XP002076660 *
KALDEN ET AL.: "NEW THERAPEUTIC APPROACHES IN AUTOIMMUNE RHEUMATIC DISEASES, WITH SPECIAL EMPHASIS ON RHEUMATOID ARTHRITIS" BRITISH JOURNAL OF RHEUMATOLOGY, Bd. 34, Nr. 3, 1995, Seiten 193-196, XP002076661 *
KERR ET AL.: "DEOXYSPERGUALIN SUPPRESSES LOCAL MACROPHAGE PROLIFERATION IN RAT RENAL ALLOGRAFT REJECTION" TRANSPLANTATION, Bd. 58, Nr. 5, 1994, Seiten 596-601, XP002076657 *
NIKOLIC-PATERSON ET AL.: "Deoxyspergualin Inhibits Mesangial Cell Proliferation and Major Histocompatibility Complex Class II Expression" J. AM. SOC. NEPHROLOGY, Bd. 5, Nr. 11, 1995, Seiten 1895-1902, XP002076656 *
PERICO ET AL.: "Prevention of Transplant Rejection" DRUGS, Bd. 54, Nr. 4, 1997, Seiten 533-570, XP002076654 *
R[IS[NEN-SOKOLOWSKI ET AL.: "Mechanism of Action of 15-Deoxyspergualin in Allograft Arteriosclerosis in Rat Aortic Transplants" TRANSPLANTATION PROCEEDINGS, Bd. 26, Nr. 5, 1994, Seite 3224 XP002076655 *
REYNOLDS: "MARTINDALE - THE EXTRA PHARMACOPOEIA. " 1996 , ROYAL PHARMACEUTICAL SOCIETY , LONDON XP002074427 224540 siehe Seite 576, Spalte 2, Absatz 3-4 *
SMITH ET AL.: "Transient immunosuppression permits successful repetitive intravenous administration of an adenovirus vector" GENE THERAPY, Bd. 3, Nr. 6, 1996, Seiten 496-502, XP002076653 *
TEPPER M.A.: "Deoxyspergualin" ANN. N.Y. ACAD. SCI., Bd. 696, 1993, Seiten 123-132, XP002076658 *
WAAGA ET AL.: "In Vitro Analysis of the Mode of Action of the Immuno-suppressive Drug 15-Deoxyspergualin" ARCHIVUM IMMUNOLOGIAE ET THERAPIAE EXPERIMENTALIS, Bd. 44, Nr. 2-3, 1996, Seiten 155-163, XP002076659 *

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Publication number Publication date
AU7034498A (en) 1998-10-20
WO1998042320A3 (fr) 1998-12-23
DE19711803A1 (de) 1998-09-24

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