ABSTRACT
The design and enzyme activities of a novel class of imidazo[2,1-b]thiazoles is presented.
Subject(s)
ErbB Receptors/antagonists & inhibitors , Protein Kinase Inhibitors/pharmacology , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Thiazoles/pharmacology , Models, Molecular , Protein Kinase Inhibitors/chemistry , Structure-Activity Relationship , Thiazoles/chemistryABSTRACT
Emerging clinical and pre-clinical data indicate that both insulin-like growth factor receptor (IGF-IR) and members of the epidermal growth factor (EGF) family of receptor tyrosine kinases (RTKs) exhibit significant cross-talk in human cancers. Therefore, a small molecule that successfully inhibits the signaling of both classes of oncogenic kinases might provide an attractive agent for chemotherapeutic use. Herein, we disclose the structure activity relationships that led to the synthesis and biological characterization of 14, a novel small molecule inhibitor of both IGF-IR and members of the epidermal growth factor family of RTKs.
Subject(s)
Antineoplastic Agents/chemical synthesis , Chemistry, Pharmaceutical/methods , ErbB Receptors/antagonists & inhibitors , ErbB Receptors/metabolism , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Receptors, Somatomedin/antagonists & inhibitors , Receptors, Somatomedin/metabolism , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Dimerization , Drug Design , Humans , Lung/metabolism , Models, Chemical , Neoplasms/metabolism , Phosphorylation , Pyrimidines/chemistry , Receptor Protein-Tyrosine Kinases/chemistry , Signal TransductionABSTRACT
Methionine aminopeptidase-2 (MetAP2) is a novel target for cancer therapy. As part of an effort to discover orally active reversible inhibitors of MetAP2, a series of anthranilic acid sulfonamides with micromolar affinities for human MetAP2 were identified using affinity selection by mass spectrometry (ASMS) screening. These micromolar hits were rapidly improved to nanomolar leads on the basis of insights from protein crystallography; however, the compounds displayed extensive binding to human serum albumin and had limited activity in cellular assays. Modifications based on structural information on the binding of lead compounds to both MetAP2 and domain III of albumin allowed the identification of compounds with significant improvements in both parameters, which showed good cellular activity in both proliferation and methionine processing assays.
Subject(s)
Aminopeptidases/chemistry , Antineoplastic Agents/chemical synthesis , Metalloendopeptidases/chemistry , Serum Albumin/chemistry , Sulfonamides/chemical synthesis , ortho-Aminobenzoates/chemical synthesis , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Cell Proliferation/drug effects , Humans , In Vitro Techniques , Mass Spectrometry , Methionine/metabolism , Models, Molecular , Protein Binding , Protein Conformation , Protein Structure, Tertiary , Rats , Structure-Activity Relationship , Sulfonamides/chemistry , Sulfonamides/pharmacology , ortho-Aminobenzoates/chemistry , ortho-Aminobenzoates/pharmacologyABSTRACT
We have screened molecules for inhibition of MetAP2 as a novel approach toward antiangiogenesis and anticancer therapy using affinity selection/mass spectrometry (ASMS) employing MetAP2 loaded with Mn(2+) as the active site metal. After a series of anthranilic acid sulfonamides with micromolar affinities was identified, chemistry efforts were initiated. The micromolar hits were quickly improved to potent nanomolar inhibitors by chemical modifications guided by insights from X-ray crystallography.
Subject(s)
Aminopeptidases/antagonists & inhibitors , Angiogenesis Inhibitors/pharmacology , Antineoplastic Agents/pharmacology , Glycoproteins/antagonists & inhibitors , Sulfonamides/pharmacology , Angiogenesis Inhibitors/chemical synthesis , Angiogenesis Inhibitors/chemistry , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Binding Sites , Cell Line , Cell Proliferation/drug effects , Crystallography, X-Ray , Drug Screening Assays, Antitumor , Humans , Manganese/chemistry , Mass Spectrometry/methods , Methionyl Aminopeptidases , Models, Molecular , Molecular Structure , Sensitivity and Specificity , Stereoisomerism , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/chemistryABSTRACT
Tumor vascularity is correlated with an aggressive disease phenotype in neuroblastoma, suggesting that angiogenesis inhibitors may be a useful addition to current therapeutic strategies. We previously showed that the antiangiogenic compound TNP-470, an irreversible methionine aminopeptidase 2 (MetAP2) inhibitor, suppressed local and disseminated human neuroblastoma growth rates in murine models but had significant associated toxicity at the effective dose. We have recently shown that a novel, reversible MetAP2 inhibitor, A-357300, significantly inhibits CHP-134-derived neuroblastoma s.c. xenograft growth rate with a treatment-to-control (T/C) ratio at day 24 of 0.19 (P < 0.001) without toxicity. We now show that the combination of A-357300 with cyclophosphamide at the maximal tolerated dose sustained tumor regression with a T/C at day 48 of 0.16 (P < 0.001) in the CHP-134 xenograft model. A-357300 also significantly inhibited establishment and growth rate of hematogenous metastatic deposits following tail vein inoculation of CHP-134 cells and increased overall survival (P = 0.021). Lastly, A-357300 caused regression of established tumors in a genetically engineered murine model with progression-free survival in five of eight mice (P < 0.0001). There was no evidence of toxicity. These data show that MetAP2 may be an important molecular target for high-risk human neuroblastomas. We speculate that the growth inhibition may be through both tumor cell intrinsic and extrinsic (antiangiogenic) mechanisms. The potential for a wide therapeutic index may allow for treatment strategies that integrate MetAP2 inhibition with conventional cytotoxic compounds.
Subject(s)
Aminopeptidases/antagonists & inhibitors , Chlorobenzenes/pharmacology , Metalloendopeptidases/antagonists & inhibitors , Neuroblastoma/prevention & control , Xenograft Model Antitumor Assays/methods , Animals , Antineoplastic Agents, Alkylating/therapeutic use , Chlorobenzenes/therapeutic use , Cyclophosphamide/therapeutic use , Drug Therapy, Combination , Humans , Mice , Mice, Inbred Strains , Mice, Nude , Mice, SCID , Mice, Transgenic , N-Myc Proto-Oncogene Protein , Neuroblastoma/pathology , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Oncogene Proteins/genetics , Oncogene Proteins/metabolism , Survival Analysis , Treatment Outcome , Tumor Cells, CulturedABSTRACT
Substituted 3-amino-2-hydroxyamides and related hydroxyamides and acylhydrazines were identified as inhibitors of human methionine aminopeptidase-2 (MetAP2). Examination of substituents through parallel synthesis and iterative structure-based design allowed the identification of potent inhibitors with good selectivity against MetAP1. Diacylhydrazine 3t (A-357300) was identified as an analogue displaying inhibition of methionine processing and cellular proliferation in human microvascular endothelial cells (HMVEC).
Subject(s)
Amides/chemistry , Amides/pharmacology , Aminopeptidases/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Metalloendopeptidases/antagonists & inhibitors , Binding Sites/drug effects , Cell Division/drug effects , Endothelium, Vascular/cytology , Endothelium, Vascular/drug effects , Humans , Methionine/drug effects , Models, Biological , Models, Molecular , Molecular StructureABSTRACT
Methionine aminopeptidase (MetAP)-2 has been suggested as a novel target for cancer therapy because the anticancer agent TNP-470 irreversibly inactivates the catalytic activity of this enzyme. However, the importance of MetAP2 in cell growth and tumor progression was uncertain because previous data were based on the chemically reactive TNP-470. Here we show that a rationally designed reversible MetAP2 inhibitor, A-357300, suppresses tumor growth preclinically without the toxicities observed with TNP-470. We have synthesized this bestatin-type MetAP2 inhibitor with the aid of crystal structures of the enzyme-inhibitor complexes and parallel synthesis. A-357300 induces cytostasis by cell cycle arrest at the G(1) phase selectively in endothelial cells and in a subset of tumor cells, but not in most primary cells of nonendothelial type. A-357300 inhibits angiogenesis both in vitro and in vivo and shows potent antitumor efficacy in carcinoma, sarcoma, and neuroblastoma murine models. These data affirm that MetAP2 plays a pivotal role in cell growth and establish that reversible MetAP2 inhibitors are promising novel cancer therapeutic agents.
