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
  • C07K16/3015—Breast
• • 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
  • A61K47/68—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6835—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
  • A61K47/6851—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
• • 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
  • A61K47/68—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6835—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
  • A61K47/6851—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
  • A61K47/6855—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 an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell the tumour determinant being from breast cancer cell
• • 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
  • A61K51/1045—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 animal or human tumor cells or tumor cell determinants
• • 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
  • A61K51/1045—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 animal or human tumor cells or tumor cell determinants
  • A61K51/1051—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 animal or human tumor cells or tumor cell determinants the tumor cell being from breast, e.g. the antibody being herceptin
• • 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
  • 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

• This invention relates to monoclonal antibodies which bind to surface antigens of human carcinoma and more particularly, relates to a monoclonal antibody which demonstrates activity with a mucin-like glycoprotein complex of very high molecular weight on the surface of breast carcinomas and some carcinomas of other tissues, but which does not bind to normal human breast epithelial cells.
• Monoclonal antibodies have been developed that recognize a high molecular weight mucin-like glycoprotein complex present on the surface of normal human breast epithelial cells. Peterson et al., Im- perial Cancer Research Fund, London, England, March 2-3 (1981); Taylor-Papadimitriou et al. , Int. J. Cancer, 28:17-21 (1981); Ceriani et al., Somatic Cell Genetics, 9:415-427 (1983). Other investigators have developed monoclonal antibodies using both the human milk fat globule as the immunizing agent [Taylor-Papadimitriou et al., Int. J.
• mucin-like glycoproteins of the human milk fat globule membrane In biochemical studies of the large molecular weight mucin-like glycoproteins of the human milk fat globule membrane, it was suggested that these glycoproteins are complexes consisting of at least three distinct components which may represent at least three distinct molecular entities. Shimizu et al., Biochem J., 233:725-730 (1986). Thus, the mucin-like glycoproteins of the human milk fat globule membrane have been shown to be large molecular complexes which, due to their size, can be expected to have a myriad of epitopes.
• Monoclonal antibodies have been developed which bind to mucin-like glycoproteins on the surface of normal breast epithelial cells and malignant breast cells such as the Mc5 monoclonal antibody described in Ceriani et al., Somatic Cell Genetics, 9:415-427 (1983). Additionally, a monoclonal antibody D-274, specific to guinea pig milk fat globule membrane, was used to determine the distribution of mucin-like glycoproteins of greater than 400,000 daltons in both benign fibrocystic disease and infiltrating duct car ⁇ cinoma of the human breast. Greenwalt et al., Am. J. Pathol., 118:351-359 (1985).
• Prior art monoclonal antibodies have been devel ⁇ oped that will bind to normal breast cells, to breast carcinomas, as well as, to some epithelial cells of other tissues.
• these monoclonal antibodies exhibit binding specificities for normal breast tissue as well as carcinoma of the human breast. It would be highly advantageous to provide a monoclonal antibody which will bind to a unique epitope that is expressed on the surface of breast carcinomas and some carcinomas of other tissues but is not expressed on normal human breast epithelial cells.
• a monoclonal antibody which binds to a novel mucin- l ike glycoprotein antigen on human breast car- cinoma cells but which does not bind normal human breast epithelial cells.
• the monoclonal antibody recognizes adenocarcinoma cells of the breast, ovary, endometrium, lung, and pancreas and will not bind normal epithelial cells of the heart, gastrointestinal tract, pituitary, prostate, mesenchymal tissues, liver, parathyroid, breast, spleen, thyroid, testis, or ovary.
• the monoclonal antibody does not bind to adenomas generally.
• the antigen recognized by the monoclonal antibody is termed "BrEl” and is characterized as having a high molecular weight which may exceed 400,000 daltons.
• the specificity of the BrEl monoclonal antibody enables ad ⁇ vantageous diagnostic and possible therapeutic applica- tions in the treatment of breast carcinomas, especially in view of the recognized great incidence of breast cancer throughout the world.
• the BrEl monoclonal antibody was developed using normal delipidated human fat globule as the immunizing agent.
• HMFG human milk fat globule
• the harvested spleen cells thereafter were fused with P3-NSl/I-Ag4-l mouse myeloma using well known polyethylene glycol techniques.
• the screening for identifying the hybridoma or hybrid cell line which produced the monoclonal antibody of the invention was done using both a solid phase radioimmuno-plate binding assay and an ELISA assay using the HMFG and components of HMFG in wells of a microtiter plate as described in Ceriani et al., Somatic Cell Genetics, 9:415-427 (1983). Thereafter, the wells positive for HMFG were screened on cell lines from human breast carcinoma tis ⁇ sue, MCF-7 [Soule et al., J. Natl.
• the HMFG was separated by a 7% polyacrylamide gel electrophoresis and then electroblotted to nitrocellulose paper.
• the nitrocel- lulose was cut into strips and the strips incubated.
• HMFG was elec- trophoresed on 7% polyacrylamide gel as described by Laemmli VK, Nature, 227:680 (1970).
• the gel lane was sliced into fractions, the slices eluted, and the eluate dried onto microtiter plates. Binding of monoclonal antibody was tested by a radioimmunobinding assay technique.
• the monoclonal antibody BrEl identified a material found only at the origin of the polyacrylamide gel.
• the mucin-like glycoprotein antigen identi ⁇ fied by the monoclonal antibody BrEl remained at the origin and did not penetrate the polyacrylamide gel.
• the HMFG then was electrophoresed on less than 7% polyacrylamide gel using the solid-phase binding assay previously described.
• the molecular weight of the mucin-like glycoprotein antigen of HMFG identified by the monoclonal antibody BrEl was then calculated using high molecular weight standards. The molecular weight of this antigen was estimated to exceed 400,000 daltons. Studies were conducted between the BrEl and Mc5 monoclonal antibodies to study the competition be ⁇ tween them for the glycoprotein antigen.
• the monoclonal antibody BrEl was found not to compete for binding with the same epitope to which the Mc5 antibody bound. In assays of body fluids which contained breast carcinoma cells or molecules using the BrEl monoclonal antibody, we determined that there occurred binding also to a glycoprotein molecular entity which exhibited a molecular weight of less than 400,000 daltons. It is postulated that the high molecular weight mucin-like glycoprotein antigen to which the BrEl monoclonal antibody bound had become denatured in the body fluid such as to fragmentize. It appeared that the BrEl monoclonal antibody recognized a common epitope on the high molecular weight antigen and the molecular entity or fragment which would explain the phenomenon.
• the BrEl monoclonal antibody binds specifical ⁇ ly to the high molecular weight antigen exceeding 400,000 daltons, it can bind to a common epitope of the antigen found on such a molecular entity. This is pos ⁇ sible since the high molecular weight antigen appears to be a complex of molecular entities.
• the BrEl monoclonal antigen isotype was determined using a mouse immunoglobulin kit to be IgG2a.
• the BrEl monoclonal antibody was tested extensive ⁇ ly for binding to histological sections in order to further characterize its tissue specificity.
• Standard immunoperoxidase assay procedures were used for binding to histological sections of normal and cancerous human tissue.
• the tissues were prepared in the form of multi-tumor tissue blocks as described by Battifora H., Lab Invest., 55:244-248 (1986). Each tissue block was prepared from a collection of strips of fixed tissues by wrapping the tissues in peritoneal membrane or in- testine and embedding in paraffin. The blocks were then sliced in preparation for the binding studies.
• the breast block used to characterize the BrEl monoclonal antibody contained 21 different specimens from normal breast, 22 adenomas, and 33 breast car ⁇ cinomas.
• the BrEl monoclonal antibody bound to the breast carcinomas but did not bind to any of the normal breast sections that were tested, nor did it bind to the adenomas. This binding occurred in spite of the fact that the source of the immunogen, HMFG, was derived from normal breast epithelial cell membrane. This observation may indicate that a transformed phenotype, or an early precursor, that is expressed in the breast carcinoma already existed in the immunizing HMFG.
• the BrEl monoclonal antibody was also tested on a large panel of different normal tissues and tumor tis- sues other than breast carcinoma. In contrast to the characterization of BrEl using the breast block, the BrEl monoclonal antibody bound to normal lung tissue and a majority of lung carcinomas. Also, it bound equally to ovarian and breast carcinomas, but it did not bind to normal ovary or breast.
• the BrEl monoclonal antibody did not stain the following tumors: nasopharyngeal, melanoma, neuroblastoma, prostate, parathyroid, peritoneum, spleen, kidney, thyroid, or embryonal. Further, the major non-breast tumors it did stain were adenocar- cinomas of the ovary, endometrium, lung, and pancreas.
• a sample of the hybrid cell capable of producing BrEl monoclonal antibodies is on deposit with the Amer ⁇ ican Type Culture Collection, Rockville, Maryland 20852, as of June 9, 1988, and is assigned A.T.C.C. No. HB 9738.
• the BrEl monoclonal antibody is unique because of its exceptional specificity for a mucin-like glycoprotein complex of very high molecular weight present on the surface of breast carcinomas and which expresses no specificity for normal breast epithelial cells. Consequently, the BrEl antibody can be espe ⁇ cially useful in the diagnosis of breast cancer in hu- mans.
• the monoclonal antibody can be tagged with a detectable label, such as a dye, fluores ⁇ cent molecules, or a radioactive tracer for tumor imag ⁇ ing.
• a suitable tracer would be Indium 111, Technetium 99 or Iodine 131.
• the BrEl monoclonal antibody may be used for therapeutic applications where conjugated to a toxin, for instance, which can lyse tumor cells to which the BrEl monoclonal antibody binds and yet not impact on normal breast epithelial tissue cells.
• a suitable radioactive label such as Indium 111, Tech ⁇ netium 99 or Iodine 131.
• Immunoassay in which microspheres are utilized in conjunction with antigens or antibodies coated thereon and suitably tagged or labeled, can be employed for in vitro diagnostic applications with the BrEl monoclonal antibody.
• the labels or tags may be varied as dis ⁇ cussed herein and known in the art.
• the BrEl antibody may be provided in an as ⁇ say kit accompanied by other ingredients for completing the assay of a biological sample according to assay in ⁇ structions in insert literature, for instance. The same may be feasible for in vivo applications, both for diagnostic and therapeutic uses.
• the assay may be used with flow cytometric procedures to study cell differen- tiation and cell-type specificity. It may also be used as a prognostic tool in the histopathology of tumors.

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• 1989
  • 1989-01-23 WO PCT/US1989/000233 patent/WO1989007268A1/en not_active Application Discontinuation
  • 1989-01-23 AU AU30491/89A patent/AU3049189A/en not_active Abandoned
  • 1989-01-23 JP JP1502149A patent/JPH03503120A/ja active Pending
  • 1989-01-23 EP EP19890902325 patent/EP0401247A4/en not_active Withdrawn
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