Preclinical Toxicity and Pharmacokinetics of the Bruton's Tyrosine Kinase-Targeting Anti-leukemic Drug Candidate, α-Cyano-β-Hydroxy-β-Methyl-N-(2,5-Dibromophenyl) Propenamide (LFM-A13)

Abstract Theleflunomide (CAS 75706-12-6) metabolite (LFM) analog α-cyano-β-hydroxy-β-methyl-N-(2,5-dibromophenyl)-propena-mide (LFM-A13, DDE-28, CAS 244240-24-2) is a rationally-designed specific inhibitor of the TEC family protein tyrosine kinase, Bruton's tyrosine kinase (BTK). LFM-A13 exhibited favorable pharmacokinetics in CD-I mice, BALB/c mice, rats, and dogs. The intraperitoneal bioavailability was estimated to be ~100%, while the oral bioavailability was ~30%. LFM-A13 enters, but does not bind to multiple tissues followed by a rapid elimination from most of the tissues. Limited distribution of LFM-A13 to extravascular tissues and its corresponding low volume of distribution could be attributed to its plasma protein binding. LFM-A13 was not toxic to mice, rats, or dogs at daily dose levels as high as 100 mg/kg. LFM-A13 formulated as a suspension and hard gelatin capsules for oral administration showed a rapid absorption and favorable pharmacokinetic features. These preclinical research studies provide the basis for future pre-IND studies and clinical development of LFM-A13 as an intravenously or orally administered new anti-leukemia agent targeting BTK.

[1]  F. Uckun,et al.  Targeting JAK3 and BTK tyrosine kinases with rationally-designed inhibitors. , 2006, Current drug targets.

[2]  J. Rosado,et al.  A role for cofilin in the activation of store-operated calcium entry by de novo conformational coupling in human platelets. , 2006, Blood.

[3]  P. Gaynon.  Childhood acute lymphoblastic leukaemia and relapse , 2005, British journal of haematology.

[4]  Ulrich Göbel,et al.  Long-term outcome in children with relapsed ALL by risk-stratified salvage therapy: results of trial acute lymphoblastic leukemia-relapse study of the Berlin-Frankfurt-Münster Group 87. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[5]  J. Rowley,et al.  Deficiency of Bruton's tyrosine kinase in B cell precursor leukemia cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[6]  G. Paré,et al.  Signaling through CD16b in human neutrophils involves the Tec family of tyrosine kinases , 2005, Journal of leukocyte biology.

[7]  G. Salido,et al.  Ca2+-independent activation of Bruton's tyrosine kinase is required for store-mediated Ca2+ entry in human platelets. , 2005, Cellular signalling.

[8]  R. Kim,et al.  Recent advances in understanding the cell death pathways activated by anticancer therapy , 2005, Cancer.

[9]  S. Rhee,et al.  Mechanism of B-Cell Receptor-Induced Phosphorylation and Activation of Phospholipase C-γ2 , 2004, Molecular and Cellular Biology.

[10]  R. Seshadri,et al.  Outcomes of unrelated cord blood transplants and allogeneic-related hematopoietic stem cell transplants in children with high-risk acute lymphocytic leukemia , 2004, Bone Marrow Transplantation.

[11]  S. Fulda,et al.  Targeting apoptosis pathways in cancer therapy. , 2004, Current cancer drug targets.

[12]  F. Uckun,et al.  Dual targeting of Bruton's tyrosine kinase and Janus kinase 3 with rationally designed inhibitors prevents graft‐versus‐host disease (GVHD) in a murine allogeneic bone marrow transplantation model , 2004, British journal of haematology.

[13]  F. Uckun,et al.  In vivo Toxicity and Antithrombotic Profile of the Oral Formulation of the Antileukemic Agent, LFM-A13-F , 2004, Arzneimittelforschung.

[14]  P. Krammer,et al.  Death receptors in chemotherapy and cancer , 2004, Oncogene.

[15]  F. Uckun,et al.  Tyrosine kinases as new molecular targets in treatment of inflammatory disorders and leukemia. , 2004, Current pharmaceutical design.

[16]  F. Uckun,et al.  Therapeutic potential of inhibiting Bruton's tyrosine kinase, (BTK). , 2004, Current pharmaceutical design.

[17]  P. Veys,et al.  Long‐term follow‐up of relapsed childhood acute lymphoblastic leukaemia , 2003, British journal of haematology.

[18]  D. Vaux,et al.  Alterations in the apoptotic machinery and their potential role in anticancer drug resistance , 2003, Oncogene.

[19]  E. Lam,et al.  BCR targets cyclin D2 via Btk and the p85α subunit of PI3-K to induce cell cycle progression in primary mouse B cells , 2003, Oncogene.

[20]  F. Uckun,et al.  The anti-leukemic Bruton's tyrosine kinase inhibitor alpha-cyano-beta-hydroxy-beta-methyl-N-(2,5-dibromophenyl) propenamide (LFM-A13) prevents fatal thromboembolism. , 2003, Leukemia & lymphoma.

[21]  M. Relling,et al.  Childhood Acute Lymphoblastic Leukaemia , 2007 .

[22]  Juhana E. Heinonen,et al.  Silencing of Bruton's tyrosine kinase (Btk) using short interfering RNA duplexes (siRNA) , 2002, FEBS letters.

[23]  M. Tolomeo,et al.  Drug resistance and apoptosis in cancer treatment: development of new apoptosis-inducing agents active in drug resistant malignancies. , 2002, Current medicinal chemistry. Anti-cancer agents.

[24]  A. Poole,et al.  Interaction of Bruton's tyrosine kinase and protein kinase Ctheta in platelets. Cross-talk between tyrosine and serine/threonine kinases. , 2002, The Journal of biological chemistry.

[25]  Scott W. Lowe,et al.  Apoptosis A Link between Cancer Genetics and Chemotherapy , 2002, Cell.

[26]  F. Uckun,et al.  A Dual Function Anti-leukemic Agent with Anti-thrombotic Activity , 2002, Leukemia & lymphoma.

[27]  F. Uckun,et al.  In vivo pharmacokinetic features, toxicity profile, and chemosensitizing activity of alpha-cyano-beta-hydroxy-beta- methyl-N-(2,5-dibromophenyl)propenamide (LFM-A13), a novel antileukemic agent targeting Bruton's tyrosine kinase. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[28]  F. Uckun,et al.  Crystal Structure of Bruton's Tyrosine Kinase Domain Suggests a Novel Pathway for Activation and Provides Insights into the Molecular Basis of X-linked Agammaglobulinemia* , 2001, The Journal of Biological Chemistry.

[29]  Fatih M. Uckun,et al.  Transcription Factor STAT5A Is a Substrate of Bruton's Tyrosine Kinase in B Cells* , 2001, The Journal of Biological Chemistry.

[30]  X. P. Liu,et al.  Structure-based design of novel anticancer agents. , 2001, Current cancer drug targets.

[31]  F. Uckun,et al.  Apoptosis Inducing Novel Anti-Leukemic Agent, Bis(4,7-Dimethyl-1,10 Phenanthroline) Sulfatooxovanadium(IV)[VO(SO4)(Me2-Phen)2] Depolarizes Mitochondrial Membranes , 2001, Leukemia & lymphoma.

[32]  H. Cavé,et al.  Long-term results of three randomized trials (58831, 58832, 58881) in childhood acute lymphoblastic leukemia: a CLCG-EORTC report , 2000, Leukemia.

[33]  F. Uckun,et al.  SPIKET and COBRA compounds as novel tubulin modulators with potent anticancer activity. , 2000, Current opinion in investigational drugs.

[34]  F. Uckun,et al.  A rationally designed anticancer drug targeting a unique binding cavity of tubulin. , 2000, Bioorganic & medicinal chemistry letters.

[35]  H. Cavé,et al.  Long-term results of three randomized trials (58831, 58832, 58881) in childhood acute lymphoblastic leukemia: a CLCG-EORTC report. Children Leukemia Cooperative Group. , 2000, Leukemia.

[36]  C. Felix,et al.  Pediatric Acute Lymphoblastic Leukemia: Challenges and Controversies in 2000. , 2000, Hematology. American Society of Hematology. Education Program.

[37]  N. Heerema,et al.  Treatment outcome and prognostic factors for infants with acute lymphoblastic leukemia treated on two consecutive trials of the Children's Cancer Group. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[38]  F. Uckun,et al.  Bruton’s Tyrosine Kinase as an Inhibitor of the Fas/CD95 Death-inducing Signaling Complex* , 1999, The Journal of Biological Chemistry.

[39]  N. Heerema,et al.  Clinical significance of Philadelphia chromosome positive pediatric acute lymphoblastic leukemia in the context of contemporary intensive therapies , 1998, Cancer.

[40]  F. Uckun,et al.  Bruton's tyrosine kinase (BTK) as a dual-function regulator of apoptosis. , 1998, Biochemical pharmacology.

[41]  S. Richards,et al.  Comparison of bone marrow transplant and chemotherapy for relapsed childhood acute lymphoblastic leukaemia: the MRC UKALL X experience , 1998, British journal of haematology.

[42]  H. Sather,et al.  Clinical features and treatment outcome of children with myeloid antigen positive acute lymphoblastic leukemia: a report from the Children's Cancer Group. , 1997, Blood.

[43]  H. Sather,et al.  Clinical features and treatment outcome of childhood T-lineage acute lymphoblastic leukemia according to the apparent maturational stage of T-lineage leukemic blasts: a Children's Cancer Group study. , 1997, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[44]  T. Kurosaki,et al.  BTK as a Mediator of Radiation-Induced Apoptosis in DT-40 Lymphoma B Cells , 1996, Science.

[45]  J. Shuster,et al.  Bone marrow transplants from HLA-identical siblings as compared with chemotherapy for children with acute lymphoblastic leukemia in a second remission. , 1994, The New England journal of medicine.

[46]  F. Uckun,et al.  Pretransplantation burden of leukemic progenitor cells as a predictor of relapse after bone marrow transplantation for acute lymphoblastic leukemia. , 1993, The New England journal of medicine.

[47]  J. Burchenal,et al.  Treatment of acute lymphoblastic leukemia. , 1972, Annual review of medicine.