WO2001019388A1 - Medicament contenant de la contactinhibine et medicament contenant un anticorps dirige contre la contactinhibine - Google Patents

Medicament contenant de la contactinhibine et medicament contenant un anticorps dirige contre la contactinhibine Download PDF

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Publication number
WO2001019388A1
WO2001019388A1 PCT/DE2000/002886 DE0002886W WO0119388A1 WO 2001019388 A1 WO2001019388 A1 WO 2001019388A1 DE 0002886 W DE0002886 W DE 0002886W WO 0119388 A1 WO0119388 A1 WO 0119388A1
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WIPO (PCT)
Prior art keywords
inhibin
contact
antibodies
antibody
contactinhibin
Prior art date
Application number
PCT/DE2000/002886
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German (de)
English (en)
Inventor
Raimund Wieser
Original Assignee
Raimund Wieser
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 Raimund Wieser filed Critical Raimund Wieser
Priority to AU74038/00A priority Critical patent/AU7403800A/en
Publication of WO2001019388A1 publication Critical patent/WO2001019388A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
    • 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
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/6415—Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins
    • 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
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/646—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies

Definitions

  • the present invention relates to a medicament which contains contact inhibin, variants or portions of contact inhibin which lead to cell lysis after induction of anti-contact inhibin antibodies, optionally in combination with a pharmaceutically acceptable carrier.
  • the present invention further relates to a medicament which contains anti-contact inhibin antibodies or fragments thereof, optionally in combination with a pharmaceutically acceptable carrier. These drugs are used in particular in tumor therapy.
  • the antigens should preferentially only occur on tumor cells, they must also be MHC-binding, peripheral T cells must recognize the presented antigens and the presented antigens must be helper T cell and cytotoxic T - Be cell activating.
  • an antigen was discovered that is very strongly expressed in 40% of all melanomas as well as in other tumors, while normal cells have only a low expression (van der Bruggen et al., Science 254 (1991), 1643). This could be used in a corresponding form as a vaccine, but a positive effect would probably only be expected in about 10% of all patients, since this antigen is HLA-Al-restricted, ie only immunogenic in combination with very specific cell surface molecules.
  • the immunogenicity of the tumor cells is increased by enabling them ex vivo by transfection of the corresponding gene to produce lymphokines (eg IL-2 or IL ⁇ -4), which after reinjection leads to local inflammation and, at least in some cases , there is a systemic reaction with tumor regression and increased T cell memory (Fearon et al., Cell 60 (1990), 397; Gansbacher et al., J.Exp. Med. 172 (1990), 1217).
  • a similar approach consists in not transferring the gene into the tumor cells themselves, but into other cells, for example cultured human fibroblasts (Veelken et al., Hum. Gen. Ther. 5 (1994), 1203-1210; Huang et al., J .Interferon cytokine Res., 15, (1995), to introduce 655-665) and then to inject, after mixing with the tumor cells.
  • the invention is essentially based on the technical problem of providing means which do not have the disadvantages of the means used in the prior art, i.e. when administered as a medicament, above all have a specific effect, that is to say essentially only influence the target cells with a change in growth, for example tumor cells.
  • This technical problem has been solved by providing the embodiments characterized in the patent claims.
  • the present invention thus relates to a medicament which contains contact inhibin, variants or portions of contact inhibin which, after administration, lead to the induction of anti-contact inhibin antibodies which cause cell lysis, optionally in combination with a pharmaceutically acceptable carrier.
  • the present invention further relates to a medicament which contains anti-contact inhibin antibodies or fragments thereof which, after administration, cause cell lysis, optionally in combination with a pharmaceutically acceptable carrier.
  • Contactinhibin is a cell membrane glycoprotein that is responsible for the cell-dependent growth regulation and thus for the homeostasis of tissues and organs. It is a precisely defined glycoprotein (60-70 kDa) with 25% N-glycosidic and 10% O-glycosidically bound oligosaccharide side chains with a pI value between 5.9 and 6.5 (Wieser et al., J. Cell Biol. 111 (1990), 2681). The N-glycosidically bound oligosaccharide side chains are probably the actual biologically active part. After binding these chains to the specific contact inhibin receptor (Gradl et al., Curr. Biol.
  • the decisive advantage over chemotherapy is that cells of the hemopoietic system or immune cells, which also have a high division rate also in the adult organism, do not express any contact inhibin molecules on the cell surface, consequently by the anti-contact inhibin antibodies produced or administered not be affected.
  • chemotherapy the destruction of the cells of the hemopoietic system or of immune cells is one of the serious side effects.
  • the antibodies can bind very efficiently to the dividing tumor cells and thus lead to cell lysis.
  • Contactinhibin is a clearly defined glycoprotein that can be isolated in its pure form, so interference occurs through contamination with other, unwanted proteins - as it e.g. the case with tumor cell extracts is - not on;
  • contactinhibin (apart from il-TMP, which only occurs on intestinal epithelial cells) is the only molecule whose causal role in the regulation of homeostasis and growth control has been clearly demonstrated.
  • N-glycans After clarifying the chemical structure of the N-glycans, they can be chemically synthesized and made available in large quantities.
  • Contact inhibin is preferably expressed in the human organism by epithelial cells.
  • the treatment is particularly successful for the main type of tumor in humans, carcinomas.
  • cells of the hematopoietic system or immune cells are not affected by therapy with contact inhibin or anti-contact inhibin antibodies.
  • Immunization with contact inhibin, variants or portions thereof will lead to the induction of anti-contact inhibin antibodies. These bind exclusively to dividing cells - preferentially to cells with uncontrolled growth, especially tumor cells - in the adult organism and trigger a complement cascade. This ultimately leads to the lysis of the cells recognized by the antibodies. The same applies if, instead of the contact inhibin for the induction of antibodies, anti-contact inhibin antibodies are already being administered. As a result, tumor cells can be eliminated very efficiently with these forms of therapy.
  • Contact inhibin can be obtained and purified, for example, as described in Example 1 below.
  • contact inhibin used here relates both to the native protein described in the prior art and to any contact inhibin variant which, compared to this form, has deletions, additions or substitutions of one or more amino acids and / or (a) modified amino acid (s) or modified oligosaccharide side chains, but is still biologically active, ie can be used to generate antibodies which lead to lysis of the desired target cell.
  • the extent to which these contact inhibin variants still have the desired biological properties can be investigated using the method described in Example 2.
  • portions of contact inhibin relates to all portions of the molecule which alone still have the biological properties discussed above.
  • These can be protein fragments, for example, or the oligosaccharide side chains originally glycosidically bound to the protein or modified forms thereof, which have a comparable effect.
  • These oligosaccharide side chains are preferably the N-glycosidically bound oligosaccaride side chains (N-glycans). These can be obtained by treating contact inhibin with N-glycanase. This enzyme cleaves the N-glycans directly from the asparagine of the amino acid chain.
  • the individual glycans are then separated and recovered using HPLC methods, with GlycoSep D and -H (from Glyco, London) being used in particular as carrier materials.
  • GlycoSep D and -H from Glyco, London
  • the glycans are labeled with the fluorescent dye ANTS (aminonaphthalinotrisulfonic acid, Glyco, London) so that it can be detected.
  • ANTS aminoonaphthalinotrisulfonic acid, Glyco, London
  • anti-contact inhibin antibody refers to any anti-contact inhibin antibody or fragment thereof which can lead to lysis of the desired dividing line in vivo. These antibodies can also be used in diagnostic assays, for example.
  • the antibodies can be monoclonal, polyclonal or synthetic antibodies or fragments thereof.
  • fragment means all parts of the monoclonal antibody (eg Fab, Fv or "Single chain Fv” fragments) which have the same epitope specificity as that have complete antibodies. The production of such fragments is known to the person skilled in the art.
  • the antibodies according to the invention are preferably monoclonal antibodies.
  • the antibodies according to the invention can be produced according to standard methods, the compounds according to the invention, for example contact inhibin, the contact inhibin receptor or parts thereof, for example short peptides, serving as immunogen.
  • Methods for obtaining monoclonal antibodies are known to the person skilled in the art.
  • the monoclonal antibody according to the invention is an antibody derived from an animal (for example a mouse), a humanized antibody or a chimeric antibody or a fragment thereof. Chimeric, human antibody-like or humanized antibodies have a reduced potential antigenicity, but their affinity for the target is not reduced.
  • the production of chimeras and humanized antibodies or of antibodies similar to human antibodies has been described in detail (see, for example, Queen et al., Proc.
  • Humanized immunoglobulins have variable scaffold regions, which essentially come from a human immunoglobulin (with the name acceptor immunoglobulin) and the complementarity of the determining regions, which essentially come from a non-human immunoglobulin (e.g. from the mouse) (with the name Donor immunoglobulin).
  • the constant region (s), if any, also originate essentially from a human immunoglobulin.
  • humanized (as well as human) antibodies offer a number of advantages over antibodies from mice or other species: (a) the human immune system should not recognize the framework or constant region of the humanized antibody as foreign, and therefore should Antibody response against such an injected antibody is lower than against a completely foreign mouse antibody or a partially foreign chimeric antibody; (b) since the Effector area of the humanized antibody is human, it interacts better with other parts of the human immune system, and (c) injected humanized antibodies have a half-life that is essentially equivalent to that of naturally occurring human antibodies, allowing smaller and less frequent ones Doses to be administered compared to antibodies of other species.
  • the present invention also relates to a hybridoma that produces the monoclonal antibody described above.
  • the anti-contact inhibin antibody or the fragment thereof is bound to a toxin in order to increase the therapeutic effect (destruction of the dividing target cell).
  • Suitable toxins and methods for coupling the toxin to the antibody are known to the person skilled in the art. Suitable toxins include, for example, the ricin A chain or a Pseudomonas toxin.
  • the medicament according to the invention is optionally in combination with a suitable pharmaceutical carrier.
  • suitable carriers and the formulation of such medicaments are known to the person skilled in the art.
  • Suitable carriers include, for example, phosphate-buffered saline solutions, water, emulsions, for example oil / water emulsions, wetting agents, sterile solutions, etc.
  • the medicament according to the invention is preferably in the form of an injection solution, the other dosage forms, such as tablets, ointments, suspensions or Emulsions are not excluded.
  • the mode of administration of the drug depends, among other things, on the form in which the active ingredient is present. It can be topical, intra-arterial (e.g.
  • the appropriate dosage is determined by the attending physician and depends on various factors, such as the age, gender, weight of the Patients, the type and stage of the disease, the mode of administration, etc.
  • Figure 1 Growth inhibition of human fibroblasts by immobilized contact inhibin
  • the specific activity was determined by determining the growth inhibition by the individual fractions in a proliferation assay according to Wieser et al. (Exp. Cell Res. 158: 493-499 (1985)).

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  • Proteomics, Peptides & Aminoacids (AREA)
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  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Molecular Biology (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Cell Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Zoology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un médicament qui contient de la contactinhibine, des variants ou des fractions de contactinhibine, qui, après leur administration, induisent la formation d'anticorps anticontactinhibine. L'invention concerne en outre un médicament qui contient des anticorps anticontactinhibine ou des fragments de ceux-ci. Ces médicaments trouvent des applications en particulier dans la thérapie des tumeurs.
PCT/DE2000/002886 1999-09-13 2000-08-23 Medicament contenant de la contactinhibine et medicament contenant un anticorps dirige contre la contactinhibine WO2001019388A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU74038/00A AU7403800A (en) 1999-09-13 2000-08-23 Medicament containing contactinhibin and medicament containing antibodies directed against contactinhibin

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19943839.0 1999-09-13
DE19943839A DE19943839A1 (de) 1999-09-13 1999-09-13 Contactinhibin enthaltendes Arzneimittel sowie Arzneimittel, das gegen Contactinhibin gerichtete Antikörper enthält

Publications (1)

Publication Number Publication Date
WO2001019388A1 true WO2001019388A1 (fr) 2001-03-22

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PCT/DE2000/002886 WO2001019388A1 (fr) 1999-09-13 2000-08-23 Medicament contenant de la contactinhibine et medicament contenant un anticorps dirige contre la contactinhibine

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AU (1) AU7403800A (fr)
DE (1) DE19943839A1 (fr)
WO (1) WO2001019388A1 (fr)

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
DATABASE CHEMABS [online] CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; SCHUETZ, SONJA ET AL: "Biosynthesis of contactinhibin a plasmamembrane glycoprotein involved in contact-dependent inhibition of growth", XP002156012, retrieved from STN Database accession no. 117:148109 HCA *
GBF MONOGR. (1991), 15(PROTEIN GLYCOSYLATION: CELL., BIOTECHNOL. ANAL. ASPECTS), 73-6 *
HEISNER ANJA ET AL: "CHARACTERIZATION OF A NOVEL CELL MEMBRANE-BOUND SIALIDASE FROM HUMAN FIBROBLASTS.", EUROPEAN JOURNAL OF CELL BIOLOGY, vol. 69, no. SUPPL. 42, 1996, pages 73, XP000971655, ISSN: 0171-9335 *
WIESER J RAIMUND ET AL: "IN VIVO MODULATED N-ACYL SIDE CHAIN OF N-ACETYLNEURAMINIC ACID MODULATES THE CELL CONTACT-DEPENDENT INHIBITION OF GROWTH.", FEBS LETTERS, vol. 395, no. 2-3, 1996, pages 170 - 173, XP002156011, ISSN: 0014-5793 *
WIESER R J ET AL: "Chemotactic migration of human diploid fibroblasts is inhibited by contactinhibin [letter].", IN VITRO CELLULAR AND DEVELOPMENTAL BIOLOGY, (1992 APR) 28A (4) 233-4., XP000971661 *
WIESER R J ET AL: "Contact-dependent inhibition of growth of normal diploid human fibroblasts by plasma membrane glycoproteins.", BIOCHIMIE, (1988 NOV) 70 (11) 1661-71., XP000971647 *
WIESER R.J. ET AL: "[Molecular basis of growth regulation by cell contact]. MOLEKULARE GRUNDLAGEN DER WACHSTUMSREGULATION DURCH ZELLKONTAKTE.", MEDIZINISCHE WELT, (1995) 46/5 (272-279)., XP000971644 *

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AU7403800A (en) 2001-04-17
DE19943839A1 (de) 2001-03-15

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