+

WO1992008489A1 - Traitement d'affections intestinales inflammatoires - Google Patents

Traitement d'affections intestinales inflammatoires Download PDF

Info

Publication number
WO1992008489A1
WO1992008489A1 PCT/US1991/008257 US9108257W WO9208489A1 WO 1992008489 A1 WO1992008489 A1 WO 1992008489A1 US 9108257 W US9108257 W US 9108257W WO 9208489 A1 WO9208489 A1 WO 9208489A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
elam1
ibd
polypeptide
elaml
Prior art date
Application number
PCT/US1991/008257
Other languages
English (en)
Inventor
Roy R. Lobb
Daniel K. Podolsky
Original Assignee
Biogen, Inc.
The General Hospital Corporation
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 Biogen, Inc., The General Hospital Corporation filed Critical Biogen, Inc.
Publication of WO1992008489A1 publication Critical patent/WO1992008489A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1027—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against receptors, cell-surface antigens or cell-surface determinants
    • 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
    • C07K16/2851—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
    • C07K16/2854—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72 against selectins, e.g. CD62
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2123/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a treatment for inflammatory bowel disease (IBD). More particularly, this invention relates to the use of antibodies recognizing endothelial cell-leukocyte adhesion molecules (ELAMs) in the treatment of IBD.
  • ELAMs endothelial cell-leukocyte adhesion molecules
  • IBD Inflammatory bowel disease
  • Ulcerative colitis is confined to the large intestine (colon) and rectum, and involves only the inner lining of the intestinal wall.
  • Crohn's disease may affect any section of the gastrointestinal tract (i.e., mouth, esophagus, stomach, small intestine, large intestine, rectum and anus) and may involve all layers of the intestinal wall. Both diseases are characterized by abdominal pain and cramping, diarrhea, rectal bleeding and fever. The symptoms of these diseases are usually progressive, and sufferers typically experience periods of remission followed by severe flareups. IBD affects an estimated two million people in the United States alone. Although IBD is not considered a fatal illness, prolonged disease can lead to severe malnutrition affecting growth or to the formation of abscesses or intestinal scar tissue, leading in turn to infection or bowel obstruction.
  • IBD has no cure, and the exact causes of IBD are not yet understood.
  • Conventional treatments for IBD have involved anti-inflammatory drugs, immunosuppressive drugs and surgery.
  • Sulfasalazine and related drugs having the bioactive 5-amino-salicylic acid (5-ASA) moiety are widely used to control moderate IBD symptoms and to maintain remission.
  • Severe inflammation is often treated with powerful corticosteroids and sometimes ACTH or with immunosuppressants such as 6-mercaptopurine and azathioprine.
  • the most common surgical treatments for severe chronic IBD are intestinal resections and, ultimately, colectomy, which is a complete cure only for ulcerative colitis. Severe side effects are associated with these drugs, including nausea, dizziness, .
  • the present invention provides novel methods for the treatment of IBD and further provides new pharmaceutical compositions useful in the treatment of IBD.
  • the present invention provides a method comprising the step of administering to an IBD sufferer an effective amount of an anti-ELAM1 antibody, such as antibody BB11 , raised against recombinant human ELAM1 expressed in COS cells.
  • the anti-ELAMl antibody is advantageously administered in vivo to a patient with active IBD and serves to inhibit neutrophil migration and the associated release of inflammatory mediators and tissue damage associated with neutrophil accumulation. It is believed that the use of anti- ELAMl antibody in the treatment of IBD will be effective to ameliorate flareups and to induce and maintain remission of IBD symptoms.
  • an immortal cell line typically myeloma cells
  • lymphocytes typically splenocytes
  • Immunization may be accomplished using standard procedures.
  • the unit dose and immunization regimen depend on the species of mammal immunized, its immune status, the body weight of the mammal, etc.
  • the immunized mammals are bled and the serum from each blood sample is assayed for particular antibodies using appropriate screening assays.
  • anti-ELAMl antibodies were identified by testing the ability of the immune serum to block HL-60 binding to IL-l ⁇ -induced human umbilical vein endothelial cells (HUVECs).
  • the lymphocytes used in the production of hybridoma cells typically are isolated from immunized mammals whose sera have already tested positive for the presence of anti-ELAMl antibodies using such screening assays.
  • the immortal cell line (e.g., a myeloma cell line) is derived from the same mammalian species as the lymphocytes.
  • Preferred immortal cell lines are mouse myeloma cell lines that are sensitive to culture medium containing hypoxanthine, aminopterin and thymidine ("HAT medium").
  • HAT-sensitive mouse myeloma cells are fused to mouse splenocytes using polyethylene glycol ("PEG") 1500.
  • Hybridoma cells resulting from the fusion are then selected using HAT medium, which kills unfused and unproductively fused myeloma cells (unfused splenocytes die after several days because they are not transformed).
  • Hybridomas producing a desired antibody are detected by screening the hybridoma culture supernatants. .
  • hybridomas prepared to produce anti-ELAMl antibodies were screened by testing the hybridoma culture supernatant for secreted antibodies having the ability to bind to a stable ELAM1-expressing cell line, such as a COS 7 cell line stably transfected with a recombinant gene for ELAM1. Supernatant antibodies were also assayed for the ability to bind cytokine-stimulated HUVECs but not unstimulated (control) HUVECs.
  • hybridoma cells that tested positive in such screening assays were cultured in a nutrient medium under conditions and for a time sufficient to allow the hybridoma cells to secrete the monoclonal antibodies into the culture medium. Tissue culture techniques and culture media suitable for hybridoma cells are well known. The conditioned hybridoma culture supernatant may be collected and the anti-ELAM1 antibodies optionally further purified by well known methods.
  • the desired antibody may be produced by injecting the hybridoma cells into the peritoneal cavity of an unimmunized mouse.
  • the hybridoma cells proliferate in the peritoneal cavity, secreting the antibody which accumulates as ascites fluid.
  • the antibody may be- harvested by withdrawing the ascites fluid from the peritoneal cavity with a syringe.
  • Anti-ELAM1 monoclonal antibodies have been previously described (Benjamin et al. 1990 [5];
  • BB11 an anti-ELAM1 monoclonal antibody designated BB11 (obtained from Biogen, Inc., Cambridge, MA) was used.
  • a hybrid cell line capable of producing BB11 is on deposit with In Vitro International, Inc.
  • Antibodies such as BB11 and other anti-ELAM antibodies capable of inhibiting adhesion of leukocytes to IBD-involved tissue could specifically inhibit migration of leukocytes to inflamed sections of the gastrointestinal tract. This inhibition of leukocyte migration across endothelium could, in turn, prevent the secondary pathological effects of leukocyte infiltration, e.g., release of toxic substances, inducement of soluble inflammatory cell mediators, etc.
  • the method of the present invention uses the specific binding activity of anti-ELAMl antibodies to reduce inflamination.
  • the anti-ELAMl antibody can be administered in the form of a pharmaceutical composition comprising an effective amount of the antibody and a pharmaceutically acceptable carrier.
  • dosages of from 0.1 mg/kg-patient/day to 2.0 mg/kg-patient/day may be used, although higher or lower dosages may be indicated with consideration to the age, sensitivity, tolerance, and other characteristics of the patient, the acuteness of the flareup, the history and course of the disease, and other similar factors • routinely considered by an attending physician.
  • dosages from 0.1 mg/kg-patient/day to 2.0 mg/kg-patient/day may be used, although higher or lower dosages may be indicated and employed with advantageous effects considering the age, sensitivity, tolerance, and other characteristics of the patient, the pattern of flareups, the history and course of the disease, and other similar factors routinely considered by an attending physician.
  • Suitable pharmaceutical carriers include, e.g., sterile saline and like solutions.
  • the pharmaceutical compositions may additionally be formulated to control the release of the active ingredients or prolong their presence in the patient's system.
  • suitable drug delivery systems include, e.g., hydrogels, hydroxmethylcellulose, microcapsules, liposomes, microemulsions, microspheres, and the like.
  • appropriately radiolabeled antibodies may also be used to monitor the course of the disease in patients.
  • anti-ELAMl antibody labeled with a radioisotope is administered to a patient suffering from IBD, the labeled antibody becomes localized at the IBD-involved tissues, and radioimaging of the IBD-involved tissues can be performed using known techniques.
  • the selection of radioisotope is based on a number of factors, e.g., toxicity, biological half-life, and detectability.
  • Preferred radioisotopes include 125 I, 123 I and 1l1 In.
  • the antibodies may be labeled by any method known in the art, such as by employing a labeling reagent (e.g.,
  • the radiolabeled antibody is administered to a patient, e.g., intravenously, and IBD-involved tissue is detected, e.g., by a radioactivity-sensitive camera coupled to computer imaging equipment. In this manner, the path of the disease, its spread or its pattern of flareup can be regularly and accurately monitored.
  • anti-ELAM1 antibodies must be effective to block leukocyte binding to ELAM1 expressed in IBD-involved tissues. Accordingly, suitable recombinant antibodies capable of blocking leukocyte adhesion may be used alternatively to naturally produced antibodies.
  • Such recombinant antibodies include antibodies produced via recombinant DNA techniques, e.g., by transforming a host cell with a suitable expression vector containing DNA encoding the light and heavy immunoglobulin chains of the desired antibody, and recombinant chimeric antibodies, wherein some or all of the hinge and constant regions of the heavy and/or the light chain of the anti-ELAM1 antibody have been substituted with corresponding regions of an immunoglobulin light or heavy chain of a different species (i.e., preferably the same species being treated for IBD, to minimize immune response to the administered antibody).
  • a different species i.e., preferably the same species being treated for IBD, to minimize immune response to the administered antibody.
  • ELAMl antibodies such as Fab, Fab'-, F(ab' ) 2 , and F(y) fragments; heavy chain monomers or dimers; light chain monomers or dimers; and dimers consisting of one heavy chain and one light chain are also contemplated herein.
  • antibody fragments may be produced by chemical methods, e.g., by cleaving an intact antibody with a protease, such as pepsin or papain, or via recombinant DNA techniques, e.g., by using host cells transformed with truncated heavy and/or light chain genes.
  • Heavy and light chain monomers may similarly be produced by treating an intact antibody with a reducing agent such as dithiothreitol or ⁇ -mercaptoethanol or by using host cells transformed with DNA encoding either the desired heavy chain or light chain or both.
  • a reducing agent such as dithiothreitol or ⁇ -mercaptoethanol
  • an ELAM1 ligand or a fragment thereof may be administered to compete for the ELAM1 binding site with the leukocyte-bound form of the ligand, thereby decreasing the recruitment of leukocytes in a similar manner to the administration of anti-ELAMl antibodies.
  • Small molecules such as oligosaccharides that mimic the binding domain of an ELAMl ligand and fit the receptor domain of ELAM1 may also be employed. (See, Devlin et al., 1990 [10], Scott and Smith, 1990 [11], and U.S.
  • compositions contemplated herein may be administered by any suitable means such as orally, intranasally, subcutaneously, intramuscularly, intravenously, intra-arterially, or parenterally. Ordinarily intravenous (i.v.) or parenteral administration will be preferred to treat flareup conditions; oral administration in a timed release vehicle will be preferred to maintain remission.
  • Human colonoscopic biopsy tissue samples were obtained, with informed consent, and prepared either as frozen sections by mounting in OCT compound and quick freezing in isopentane/liquid nitrogen or as paraffin sections by fixing in 4% formaldehyde and embedding in paraffin. Frozen sections were also prepared from cotton-top tamarin colon samples.
  • the human colon samples were from normal colon, active ulcerative colitis colon (UC-active), inactive ulcerative colitis colon (UC-inactive), and uninvolved ulcerative colitis colon, as well as biopsies of colon polyps and samples of cancer-involved ulcerative colitis colon (Cancer/UC) .
  • the cotton-top tamarin samples were of active colitis, inactive colitis and intestinal tumor tissues.
  • Frozen sections (6 ⁇ ) placed on gelatin-coated slides (1% gelatin, heated at 60[ 1-2 min. , air dried, 1% formaldehyde, room temp., air dried) were air dried, fixed in acetone for five to ten minutes, and air dried again. Paraffin sections were prepared for testing by cutting thin sections ( ⁇ 6 ⁇ ) with a microtome and mounting on slides.
  • the excess fluid was blotted, and monoclonal antibody BB11 , which recognizes ELAM1 ; monoclonal antibody 4B9, recognizing VCAM1 ; and an anti-bovine serum albumin (anti-BSA) antibody (negative control) were applied to separate samples of each type of tissue as follows: The antibodies were diluted 1:25 in phosphate buffered saline/1% fetal bovine serum/0.1% sodium azide and the solution applied to cover the samples on each slide for 60 minutes.
  • the samples were then washed twice in phosphate buffered saline/0.2% gelatin and exposed to peroxidase- conjugated rabbit anti-mouse immunoglobulins (Dako Corp., Santa Barbara, CA) in phosphate buffered saline/1% fetal bovine serum/0.1% sodium azide/1% baboon serum for 60 minutes.
  • the samples were then washed as before, then exposed to 0.25 mg/ml 3-amino- 9-ethylcarbazole (Aldrich Chemical Co., Milwaukee, WI) in 2% N,N-dimethylformamide/0.1M acetate buffer (pH 5.2) with 0.08 hydrogen peroxide for six minutes.
  • the samples were washed with water and counterstained with 1% methyl green.
  • the anti-ELAMl and anti-VCAM1 antibodies were obtained from Biogen, Inc. (Cambridge, MA).
  • Anti-BSA antibody was obtained commercially from Sigma Chemical Co. (St. Louis, MO).
  • each preparation is calculated based on the weight of the cotton-top tamarins to provide approximately 2 mg/kg antibody per animal.
  • the preparations are administered by i.v. injection CTTs grouped as follows: Group Dose Schedule
  • a 1 dose/day of BB11 for 14 days B 1 dose/day of IgG (control) for 14 days
  • Serum samples are taken from each animal at 0, 1, 12, 24 hours and daily thereafter until 3-5 days after the last injection to determine average serum levels of antibody and half-life.
  • Biopsies of colon tissue are taken at the same times to assess leukocyte infiltration, Ig coating of the CTT endothelium, and possible vasculitis during the course of therapy.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Optics & Photonics (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Procédé de traitement d'affections intestinales inflammatoires. Le procédé consiste à administrer un anticorps, un polypeptide ou une autre molécule reconnaissant les ELAM1 (molécules d'adhésion de cellules endothéliales-leucocytes), une protéine induite à la surface de cellules endothéliales et dont on a découvert qu'elle est associée aux tissus intestinaux impliqués dans des affections intestinales inflammatoires.
PCT/US1991/008257 1990-11-08 1991-11-06 Traitement d'affections intestinales inflammatoires WO1992008489A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US61051290A 1990-11-08 1990-11-08
US610,512 1990-11-08

Publications (1)

Publication Number Publication Date
WO1992008489A1 true WO1992008489A1 (fr) 1992-05-29

Family

ID=24445320

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1991/008257 WO1992008489A1 (fr) 1990-11-08 1991-11-06 Traitement d'affections intestinales inflammatoires

Country Status (2)

Country Link
AU (1) AU9055091A (fr)
WO (1) WO1992008489A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993000908A1 (fr) * 1991-07-06 1993-01-21 Antisoma Limited Ligands de recepteur d'elam-1 utilises comme composes diagnostiques
EP0759769A4 (fr) * 1994-05-11 1998-01-28 Affymax Tech Nv Peptides et composes se liant aux selectines notamment du type elam-1

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0314863A2 (fr) * 1987-11-02 1989-05-10 Baylor College Of Medicine Utilisation du ICAM-1 ou de ses derivés fonctionels pour le traitement des inflammations non spécifiques
WO1990005539A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Anticorps specifiques contre elam-1 et leur utilisation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0314863A2 (fr) * 1987-11-02 1989-05-10 Baylor College Of Medicine Utilisation du ICAM-1 ou de ses derivés fonctionels pour le traitement des inflammations non spécifiques
WO1990005539A1 (fr) * 1988-11-14 1990-05-31 Brigham And Women's Hospital Anticorps specifiques contre elam-1 et leur utilisation

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Biochemical and Biophysical Research Communications, vol. 171, no. 1, 31 August 1990, C. BENJAMIN et al.: "A blocking monoclonal antibody to endothelial-leukocyte adhesion molecule-1 (ELAM1)", pages 348-353, see the whole document *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993000908A1 (fr) * 1991-07-06 1993-01-21 Antisoma Limited Ligands de recepteur d'elam-1 utilises comme composes diagnostiques
US5786322A (en) * 1992-05-06 1998-07-28 Affymax Technologies N.V. Peptides and compounds that bind selectins including endothelium leukocyte adhesion molecule 1
EP0759769A4 (fr) * 1994-05-11 1998-01-28 Affymax Tech Nv Peptides et composes se liant aux selectines notamment du type elam-1

Also Published As

Publication number Publication date
AU9055091A (en) 1992-06-11

Similar Documents

Publication Publication Date Title
EP0625912B1 (fr) Traitement de l'inflammation intestinale
US5223426A (en) Monoclonal antibodies reactive with defined regions of the t-cell antigen receptor
US7658923B2 (en) Anti-CD26 monoclonal antibodies as therapy for diseases associated with cells expressing CD26
Rasmussen et al. Lymphocyte recognition of lymph node high endothelium. VII. Cell surface proteins involved in adhesion defined by monoclonal anti-HEBFLN (A. 11) antibody.
US4675287A (en) Monoclonal antibody directed to human ganglioside GD2
US4628027A (en) Vitro diagnostic methods using monoclonal antibodies against connective tissue proteins
AU621304B2 (en) Method of reducing tissue damage at an inflammatory site using a monoclonal antibody
US5510466A (en) Scavenger receptor protein and antibody thereto
JP3541296B2 (ja) 腫瘍血管内皮細胞に特異的に結合するモノクローナル抗体とその利用方法
EA001200B1 (ru) Фармацевтическая композиция, способ лечения, способ ингибирования иммунного ответа
US5403919A (en) Method to control leukocyte extravasation
US8093010B2 (en) Angiogenesis inhibiting molecules, their selection, production and their use in the treatment of cancer
DE69431167T2 (de) Verfahren zum selektiven methioninentzug für maligne säugetierzellen
CA1251729A (fr) Methode de diagnostic in vitro faisant appel a des anticorps monoclonaux contre des proteines du tissu conjonctif
WO1992008489A1 (fr) Traitement d'affections intestinales inflammatoires
US5055559A (en) Anti-melanoma antibody MG-21 for diagnosis and therapy of human tumors
WO1992014757A1 (fr) Molecules d'adhesion cellule endotheliale-monocyte
Rosen et al. Monoclonal antibodies: their promise for tumor diagnosis, staging, and therapy
Stohl et al. A functionally unique anti-CD3 monoclonal antibody cross-reactive with basal keratinocytes
IL108741A (en) Active monoclonal antibodies with defined regions of the T cell antibody receptor
CA2005121A1 (fr) Reactif d'anticorps monoclonaux avec des regions definies des recepteurs d'antigenes des cellules t

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP KR NO

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载