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WO1990010692A1 - Anticorps monoclonal utilise pour la detection et le traitement de la leucemie infantile - Google Patents

Anticorps monoclonal utilise pour la detection et le traitement de la leucemie infantile Download PDF

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Publication number
WO1990010692A1
WO1990010692A1 PCT/US1990/001392 US9001392W WO9010692A1 WO 1990010692 A1 WO1990010692 A1 WO 1990010692A1 US 9001392 W US9001392 W US 9001392W WO 9010692 A1 WO9010692 A1 WO 9010692A1
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Prior art keywords
cells
monoclonal antibody
cell line
antibody
nalm
Prior art date
Application number
PCT/US1990/001392
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English (en)
Inventor
William F. Cassano
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University Of Florida
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Publication date
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Publication of WO1990010692A1 publication Critical patent/WO1990010692A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • 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
    • A61K47/6867—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 a cell of a blood cancer
    • 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
    • C07K16/3061—Blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57426—Specifically defined cancers leukemia
    • 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 currently used technique involves a monoclonal antibody or a cocktail of antibodies against tumor surface proteins with which the isolated bone marrow is incubated.
  • Monodisperse polystyrene microspheres 4.5 um in size, with covalently attached sheep anti-mouse IgG (e.g., Dynal M450, Dynal Inc., Great Neck, NY) are then added to the mixture in order to bind to the tumor cell-attracted monoclonal antibodies (MoAbs) (rosette formation).
  • the magnetic cells are removed by strong magnets and the clean bone marrow is re-injected into the patient.
  • the effectiveness of ABMT procedures depends upon several factors including the type of beads used, how the beads are coated, and the nature of the antibodies used to coat the beads.
  • the antibodies used it is advantageous to have antibodies which bind only to the target cells and do not remove desirable cells. Also, the binding to the target cells must be of a nature that a high proportion of these target cells are removed.
  • the antibodies and mixtures of antibodies which have been used to coat magnetic microspheres used in bone marrow purging systems have had moderate success. Lack of selectivity and low rates of removal of target cells have limited the effectiveness of these procedures.
  • the subject invention concerns a novel monoclonal antibody (MoAb), the cell line used to produce this MoAb, and magnetic microspheres coated with the novel MoAb.
  • MoAb monoclonal antibody
  • the MoAb of the subject invention recognizes the common acute lymphocytic leukemia (ALL) antigen (CALLA, CD10, BgplOO).
  • ALL common acute lymphocytic leukemia
  • This novel antibody is a high affinity murine IgGl, kappa (IgGlk) antibody which can be used in the diagnosis and identification of childhood leukemia.
  • the novel antibody which has been designated WCMH15.14, can also be used in the therapy of childhood leukemia.
  • the therapy involves immunomagnetic purging of residual leukemic cells in preparation for autologous bone marrow transplantation in children with acute lymphocytic leukemia.
  • the monoclonal antibody may also find therapeutic application in hybrid toxin systems where the MoAb is coupled with a toxin such as diphtheria or reicin. These systems selectively seek out and destroy the target cancer cells.
  • the monoclonal antibody of the subject invention is a murine IgGlk antibody produced from the fusion of mouse spleen oocytes immunized with the Nalm-6 cell line and fused to the NS1 myeloma cell line. Successful hybrids were selected in HAT median and cloned by limiting dilution on thymocyte feeder layers.
  • the fusion which ultimately produced the MoAb possessing the advantageous features described here resulted in approximately 2000 successful hybrids.
  • the production of hybrids is a very random event; no two hybrids are alike, and it is impossible to predict or control all the relevant characteristics of the resulting hybrids.
  • the 2000 successful hybrids resulting from this fusion only 80 were found to produce antibody which bound to Nalm-6. Of those 80 hybrids producing the desired antibody, less than half were found to have the desired selectivity. That is, 37 of the hybrids produced an antibody which bound to Nalm-6 but not Jurkat cells. Even in the unlikely event that a cell line produces the desired antibody and has the necessary selectivity, the cell line may be of no value if it is not stable.
  • the subject culture deposit will be stored and made available to the public in accord with the provisions of the Budapest Treaty for the Deposit of Microorganisms, i.e., it will be stored with all the care necessary to keep it viable and uncontaminated for a period of at least five years after the most recent request for the furnishing of a sample of the deposits, and in any case, for a period of at least 30 (thirty) years after the date of deposit or for the enforceable life of any patent which may issue disclosing the culture.
  • the depositor acknowledges the duty to replace the deposit should the depository be unable to furnish a sample when requested, due to the condition of the deposit. All restrictions on the availability to the public of the subject culture deposit will be irrevocably removed upon the granting of a patent disclosing it.
  • the monoclonal antibody produced by the deposited cell line has several uses in the detection and therapy of children with acute lymphocyte leukemia.
  • the antibody can be used in standard immunoassay procedures in order to detect the presence of leukemia antigens in a biological sample suspected of containing evidence of the disease.
  • WCMH15.14 MoAb can be used in the treatment of leukemia patients is the coating of magnetic microbeads which are to be used in the purging of bone marrow from these patients.
  • the characteristics of a monoclonal antibody that would be most useful for clinical use in purging marrow of residual leukemic cells prior to autologous marrow transplants for children with acute lymphoblastic leukemia would include: (1) the ability of the antibody to bind to the leukemic cells in a high percentage of patients, (2) the ability of the antibody to efficiently purge leukemic cells from marrow when used with magnetic microspheres in the immunomagnetic purging procedure, and (3) the antibody should not bind to stem cells which are needed to reconstitute normal marrow function after transplantation.
  • the monoclonal antibody of the subject invention binds target cells selectively, and with a high degree of affinity, WCMH15.14, or immunological equivalents, can be used alone or in combination with a cocktail of other MoAbs to increase the efficiency of the bone marrow purging system.
  • microspheres can be used to coat the beads using standard procedures well known to those skilled in the art. See, for example, Treleaven et al. (1984), supra: Kemshead et al. (1986), supra: Kvalheim, G., O. Fodstad, A. Pihl, K.
  • Another procedure which utilizes the novel monoclonal antibody of the subject invention in the therapy of leukemia patients is the selective destruction of target cells with appropriate toxins.
  • This technique involves the coupling of the novel MoAbs to a toxin so that the toxin is specifically delivered to the target cells.
  • This procedure can be used in vivo and the antibody coupled to a number of different toxins, including poisonous lectins, ricin, abrin, modeccin, botulina, and diphtheria toxin.
  • the construction of such a hybrid toxin could proceed, for example, according to the disclosures of United States Patent Nos. 4,675,382
  • the cells were plated out into four 96-well flat bottom plates with approximately 200 microliters per well. The plating was done with a 10 cc pipette to minimize shearing forces of the newly formed hybrids. The four plates were then incubated at 37°C with 5% C0 2 in air for 10 days, at which time vigorous growth in each well was apparent and beginning to exhaust the nutrient supply.
  • the cells were washed 3 times with cold PBS, reacted with a goat anti-mouse peroxidase conjugated antibody for 30 minutes, washed twice again, and the peroxidase substrates o-phenylenediamine and hydrogen peroxide were added.
  • the brown color indicating a positive reaction identified the wells containing monoclonal antibody binding to the particular cell.
  • the results of this screen indicated that 80 wells contained antibody reacted with Nalm-6 cells (20% of the 400 wells) and that 37 wells contained antibody that bound to Nalm-6 cells but not to Jurkat cells.
  • the remainder of the clones are stable and underwent extensive characterization with a variety of immunoassays, as well as immunoprecipitation experiments involving 1-125 surface labeled Nalm-6 cells.
  • One antibody, WCMH15.14 precipitated a 100 kilodalton surface glycoprotein, blocked the biotinylated CA LA antibody, and exhibited the characteristic cell binding distribution of other CD10 antibodies.
  • the other 11 stable clones do not bind the common ALL antigen.
  • the antibodies were isotyped using the Zymed mouse isotyping kit.
  • WCMH15.14 was found to be a murine IgGl, kappa monoclonal antibody.
  • Purging experiments were conducted where magnetic microbeads were coated with a variety of monoclonal antibodies to test the ability of these antibody-coated beads to bind to the target cells and to assess the selectivity and range of the binding.
  • Cells from the Na_m-6 pre-B cell line were used as representative leukemia cells. 1 x 10 6 cells per well were placed in a 96-well microtiter plate and cooled to 4°C. One hundred ml of a 1:200 dilution of the selected monoclonal antibody in phosphate buffered saline (PBS) was incubated at 4°C for 30 minutes. The cells were then washed 3 times with PBS and transferred in cold PBS to a 12x75 mm plastic tube containing 4 ml of PBS.
  • PBS phosphate buffered saline
  • WCMH15.14 can be expected to have a high probability of effectively removing leukemic cells for a broad range of patients suffering from childhood leukemia.
  • the ability to bind with leukemia cells from a high percentage of the patients tested also has important implications in terms of the antibody's use as a diagnostic tool. Because it recognizes a broad range of leukemic cells, WCMH15.14 would be less likely to give false negatives when used to detect the presence of leukemic cells in a biological sample.
  • Example 2 Use of WCMH15.14 in Cocktail with Other Monoclonal Antibodies
  • monoclonal antibodies that recognize malignant and normal pre-B cells have been shown to be useful in a cocktail along with WCMH15.14 to purge leukemic bone marrow.
  • These antibodies include DU- A L-1, a CD9 antibody, and HD37, a CD19 antibody.
  • CD9 and CD19 antibodies, as well as other unclustered antibodies may also be useful in this purging process in combination with WCMH15.14.
  • Antibody WCMH15.14 along with other CALLA antibodies have been used to phenotype malignant cells from patients with leukemia and lymphomas. It has been clearly established through many published studies (see, for example,

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  • Health & Medical Sciences (AREA)
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Abstract

L'invention a trait à une nouvelle lignée cellulaire ainsi qu'à l'anticorps monoclonal produit par ladite lignée cellulaire. Ledit nouvel anticorps monoclonal se lie avantageusement et spécifiquement à l'antigène de la leucémie lymphotique aiguë. On peut par conséquent utiliser cet anticorps pour la détection de la leucémie infantile ainsi que dans des interventions thérapeutiques telles que la purge de moëlle osseuse. On peut aussi utiliser le nouvel anticorps monoclonal pour former des protéines de toxines hybrides capables de chercher et d'éliminer sélectivement des cellules cibles.
PCT/US1990/001392 1989-03-15 1990-03-14 Anticorps monoclonal utilise pour la detection et le traitement de la leucemie infantile WO1990010692A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US32364689A 1989-03-15 1989-03-15
US323,646 1989-03-15

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WO1990010692A1 true WO1990010692A1 (fr) 1990-09-20

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994007138A1 (fr) * 1992-09-14 1994-03-31 Fodstad Oystein Detection de cellules cibles specifiques dans une population de cellules specialisees ou mixtes et solutions contenant des populations de cellules mixtes
EP0671950A1 (fr) * 1992-10-19 1995-09-20 Dana Farber Cancer Institute Purge immunologique de cellules tumorales de la moelle osseuse a l'aide de microspheres et d'anticorps monoclonaux
US6265229B1 (en) 1994-03-10 2001-07-24 Oystein Fodstad Method and device for detection of specific target cells in specialized or mixed cell populations and solutions containing mixed cell populations
US7198787B2 (en) 1996-03-13 2007-04-03 Oystein Fodstad Method of killing target cells in harvested cell populations with one or more immuno-toxins
JP2007302696A (ja) * 1993-03-19 2007-11-22 Speywood Lab Ltd 細胞活性を制御するための新規作用薬

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Cancer Research, Volume 47, 1 February 1987, (Philadelphia, PA, US), G. KVALHEIM et al.: "Elimination of B-Lymphoma Cells from Human Bone Marrow: Model Experiments using Monodisperse Magnetic Particles Coated with Primary Monoclonal Antibodies", see pages 846-851 *
Cancer Research, Volume 48, 15 August 1988, (Philadelphia, PA, US), H. HARA et al.: "Efficient Transplantation of Human Non-T-Leukemia Cells into Nude Mice and Induction of Complete Regression of the Transplanted Distinct Tumors by Ricin A-Chain Conjugates of Monoclonal Antibodies SN5 and SN6", see pages 4673-4680 *
Eur. J. Cancer, Volume 37, 1986, (Oxford, GB), K. SUGITA et al.: "Use of a Cocktail of Monoclonal Antibodies and Human Complement in Selective Killing of Acute Lymphocytic Leukemia Cells", see pages 351-357 *
Experimental Hematology, Volume 17, No. 8, August 1989, (New York, US), C.J. BIEVA et al.: "Malignant Leukemic Cell Separation by Iron Colloid Immunomagnetic Adsorption", see pages 914-920 *
Proc. Natl. Acad. Sci. USA, Volume 79, July 1982, (Washington, DC, US), R. UEDA et al.: "Serological Analysis of Cell Surface Antigens of Null Cell Acute Lymphocytic Leukemia by Mouse Monoclonal Antibodies", see pages 4386-4390 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994007138A1 (fr) * 1992-09-14 1994-03-31 Fodstad Oystein Detection de cellules cibles specifiques dans une population de cellules specialisees ou mixtes et solutions contenant des populations de cellules mixtes
US6184043B1 (en) 1992-09-14 2001-02-06 FODSTAD øYSTEIN Method for detection of specific target cells in specialized or mixed cell population and solutions containing mixed cell populations
US6893881B1 (en) 1992-09-14 2005-05-17 Abbott Laboratories, Inc. Method for detection of specific target cells in specialized or mixed cell population and solutions containing mixed cell populations
USRE43979E1 (en) 1992-09-14 2013-02-05 Abbott Laboratories Method for detection of specific target cells in specialized or mixed cell population and solutions containing mixed cell populations
EP0671950A1 (fr) * 1992-10-19 1995-09-20 Dana Farber Cancer Institute Purge immunologique de cellules tumorales de la moelle osseuse a l'aide de microspheres et d'anticorps monoclonaux
EP0671950A4 (fr) * 1992-10-19 1996-07-31 Dana Farber Cancer Inst Inc Purge immunologique de cellules tumorales de la moelle osseuse a l'aide de microspheres et d'anticorps monoclonaux.
JP2007302696A (ja) * 1993-03-19 2007-11-22 Speywood Lab Ltd 細胞活性を制御するための新規作用薬
JP4740206B2 (ja) * 1993-03-19 2011-08-03 イプセン ディベロプメンツ リミテッド 細胞活性を制御するための新規作用薬
US6265229B1 (en) 1994-03-10 2001-07-24 Oystein Fodstad Method and device for detection of specific target cells in specialized or mixed cell populations and solutions containing mixed cell populations
US7198787B2 (en) 1996-03-13 2007-04-03 Oystein Fodstad Method of killing target cells in harvested cell populations with one or more immuno-toxins

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