asta-z-7557 and Leukemia--Lymphoid

Topics: Adolescent; Adult; Animals; Bleomycin; Bone Marrow; Bone Marrow Transplantation; Cell Separation; Child; Child, Preschool; Clinical Trials as Topic; Cyclophosphamide; Drug Evaluation; Etoposide; Graft vs Host Disease; Hematopoietic Stem Cells; Humans; Leukemia; Leukemia, Lymphoid; Lymphoma; Lysophosphatidylcholines; Methylprednisolone; Neoplasms, Experimental; Phospholipid Ethers; Pyrimidinones; Transplantation, Autologous; Transplantation, Isogeneic

Topics: Adolescent; Adult; Animals; Bleomycin; Bone Marrow; Bone Marrow Transplantation; Cell Separation; Child; Child, Preschool; Clinical Trials as Topic; Cyclophosphamide; Drug Evaluation; Etoposide; Graft vs Host Disease; Hematopoietic Stem Cells; Humans; Leukemia; Leukemia, Lymphoid; Lymphoma; Lysophosphatidylcholines; Methylprednisolone; Neoplasms, Experimental; Phospholipid Ethers; Pyrimidinones; Transplantation, Autologous; Transplantation, Isogeneic

Topics: Adult; Bone Marrow; Bone Marrow Transplantation; Cell Separation; Child; Clinical Trials as Topic; Cyclophosphamide; Etoposide; Humans; Leukemia, Lymphoid; Transplantation, Autologous

Leukemia cells were mixed with normal human bone marrow cells to simulate bone marrow from leukemia patients; the mixture was then treated with a combination of stabilized derivative of cyclophosphamide [Mafosfamide (ASTA Z 7557)] and pokeweed antiviral protein-containing immunotoxin. The ability of this protocol for selective elimination of B-ALL cells was evaluated by clonogenic assay. The monoclonal antibody (B43) portion of the immunotoxin was directed against human B-cells and was linked to pokeweed antiviral protein by a disulfide bond. The combination of ASTA Z 7557 and immunotoxin was superior to either ASTA Z 7557 or the immunotoxin alone and produced nearly 7 logs of elimination of leukemia cells from the cell mixtures. About 5 logs of contaminating tumor cells were eliminated from a 200-fold excess of normal marrow under conditions where fewer than 50% of pluripotent stem cells were lost. Moreover, this manipulation did not inhibit subsequent production of pluripotent stem cells in long-term bone marrow cultures, indicating that the more primitive progenitors were not harmed.

Topics: Antibodies, Monoclonal; Antiviral Agents; B-Lymphocytes; Bone Marrow Cells; Cell Line; Cell Survival; Cells, Cultured; Chloroquine; Cyclophosphamide; Drug Synergism; Flow Cytometry; Humans; Leukemia, Lymphoid; Lymphoma; N-Glycosyl Hydrolases; Plant Proteins; Ribosome Inactivating Proteins, Type 1; Tumor Stem Cell Assay

ASTA Z 7557 potentiated the ex vivo efficiency of a T-cell directed immunotoxin containing pokeweed antiviral protein (PAP). We used an immunotoxin of pan-T monoclonal antibody 3-A1 directed against p41 antigen expressed both on normal and leukemic T-cells. Treatment with 3A1-PAP in combination with ASTA Z 7557 produced 7-8 logs elimination of target lymphoblastic leukemia cells. Our data suggest that this new strategy shows potential for more effective ex vivo marrow purging in autologous marrow transplantation for acute lymphoblastic leukemia.

Topics: Antibodies, Monoclonal; Antibody Specificity; Antibody-Dependent Cell Cytotoxicity; Bone Marrow Transplantation; Cell Line; Cyclophosphamide; Humans; Leukemia, Lymphoid; Neoplasm Proteins; T-Lymphocytes

We evaluated a novel ex vivo "purging" protocol for selective elimination of neoplastic T cells from human marrow by using a sensitive clonogenic assay. Immunotoxins (IT) were synthesized by conjugating ricin (R) to four different monoclonal antibodies (MoAb) directed against distinct markers of T cell lineage. Treatment with anti-p67-R produced effective elimination of leukemic T cells from human marrow. The cyclophosphamide congener mafosfamid (ASTA Z 7577) markedly enhanced the target cell cytotoxicity of IT and extended the final level of clonogenic kill 2 to 3 logs. Our data show that anti-p67-R in combination with mafosfamid resulted in a maximum elimination of 6.2 logs of neoplastic T cells with minimal toxicity to normal bone marrow progenitors. The efficiency of this protocol was not reduced in the presence of excess normal bone marrow cells. Similar findings were obtained by using a cocktail of four different anti-T cell IT. This approach is unique in combining both immunologic (IT) and chemical (mafosfamid) strategies for more effective ex vivo bone marrow purging in autologous bone marrow transplantation for T cell acute lymphoblastic leukemia/lymphoblastic lymphoma.

Topics: Antibodies, Monoclonal; Antineoplastic Combined Chemotherapy Protocols; Bone Marrow; Bone Marrow Transplantation; Burkitt Lymphoma; Cell Line; Cyclophosphamide; Growth Inhibitors; Humans; Leukemia, Lymphoid; Lymphocyte Depletion; Ricin; T-Lymphocytes; Tumor Stem Cell Assay

Studies were performed to evaluate the anti-tumor activity of mafosfamid, a new synthetic derivative of cyclophosphamide. We tested its ability to eliminate lymphoblastic leukemia cells from autologous bone marrow grafts following a 30 min preincubation in a highly sensitive clonogenic assay. Treatment with 50-100 micrograms mafosfamid/ml eliminated more than 4 logs of contaminating clonogenic tumor cells from a 200-fold excess of normal bone marrow. Flow cytometric studies showed differences in cell cycle kinetics between mafosfamid-resistant and mafosfamid-susceptible tumor cell clones. Compared to drug susceptible clonogenic tumor cells, clones that resisted treatment with 100 micrograms mafosfamid/ml exhibited a smaller percentage of cells in S-phase, indicating that mafosfamid is mostly cytotoxic to rapidly cycling tumor cells. The combination of mafosfamid and a target cell selective immunotoxin containing pokeweed anti-viral protein was superior to mafosfamid alone or immunotoxin alone for purging mafosfamid-resistant leukemic cells from human marrow.

Topics: Bone Marrow; Bone Marrow Transplantation; Cyclophosphamide; DNA, Neoplasm; Drug Resistance; Hematopoietic Stem Cells; Humans; Kinetics; Leukemia, Lymphoid; Tumor Stem Cell Assay

Topics: Adolescent; Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Bone Marrow; Bone Marrow Transplantation; Cell Separation; Combined Modality Therapy; Cyclophosphamide; Follow-Up Studies; Humans; Leukemia, Lymphoid; Middle Aged; Transplantation, Autologous

The contamination of autologous marrow with clonogenic tumor cells has been the main argument against ABMT in acute leukemia. In a preclinical study we evaluated an active cyclophosphamide derivative named "ASTA Z 7557". We observed that the toxic effect of this drug on CFU-GM growth was dependent on nucleated cell concentration as well as on red blood cell contamination. The potency of the drug was in close relationship with the incubation temperature. The growth of leukemic CFU was inhibited with an ASTA Z dose higher than 30 micrograms/ml. In our system, beyond 40 micrograms/ml more than 95% of committed stem cells are destroyed. Fifteen patients had autotransplant because of AML for 10 patients and because of ALL for 5 patients (4 patients were grafted in relapse and 11 patients in remission). We demonstrated that the marrow take was possible although the inoculum is CFU-GM depleted. Five of the 10 AML patients are alive and remain disease-free at 45+, 65+, 190+, 345+ and 570+ days from ABMT without any maintenance treatment. Four of the 5 ALL patients are alive, three of them in complete remission (40+, 110+, 250+ days). The number of patients reported in this clinical study was relatively small and more cases should be evaluated to be conclusive. Nevertheless the feasibility of chemopurified ABMT was demonstrated.

Topics: Adult; Bone Marrow Transplantation; Cell Division; Cell Separation; Child; Child, Preschool; Colony-Forming Units Assay; Cyclophosphamide; Drug Evaluation; Female; Hematopoietic Stem Cells; Humans; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Male; Neoplastic Stem Cells; Stem Cells; Transplantation, Autologous