Peptide and peptide mimetic binding antagonists of polo-like kinase 1 polo box domain and methods of use
Inventors
Burke, Jr., Terrence R. • Hymel, David T. • TSUJI, Kohei
Assignees
US Department of Health and Human Services
Publication Number
US-10905769-B2
Publication Date
2021-02-02
Expiration Date
2035-11-13
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Abstract
The description provides novel compounds that may serve as anticancer therapeutics. The compounds of the description bind to polo-like kinases through the polo-box domain. The peptide derivatives of the description have achieved improved efficacy in biochemical assays against Plk1. Exemplary compounds of the description include macrocyclic peptidomimetics with high affinity and selectivity for polo-like kinases, which may provide the basis for a new genre of anticancer therapeutics. Other exemplary compounds of the description include bi-valent compounds with that bind to polo-like kinases through both kinase domain and polo-box domain simultaneously by incorporating additional moieties that target Plk1 kinase domain, which significantly enhances affinitity relative and may provide the basis for a new genre of anticancer therapeutics. The description also provides methods of use, methods of preparation, compositions, and kits thereof. Further, the description provides a novel method of design and/or synthesis of phosphoryl-derived peptide derivatives useful as therapeutic agents.
Core Innovation
The present disclosure provides novel compounds, particularly peptide derivatives and peptidomimetics, that inhibit polo-like kinases, especially Polo-like kinase 1 (Plk1), by binding to the polo-box domain (PBD). These compounds include macrocyclic peptidomimetics with high affinity and selectivity for polo-like kinases, and bi-valent compounds capable of binding both the kinase domain (KD) and PBD simultaneously, leading to significantly enhanced affinity. The use of the phosphonate analog, (2S,3R)-2-amino-3-methyl-4-phosphonobutanoic acid (Pmab), confers phosphatase stability, preventing inactivation by cellular phosphatases, which is a limitation of phosphothreonine-containing ligands.
The problem addressed arises from the critical role of Plk1 in cell proliferation and tumorigenesis and the need for selective inhibitors to avoid off-target effects associated with ATP-competitive kinase domain inhibitors. PBD-directed antagonists provide a selective alternative due to unique PBD presence in Plk family kinases. However, existing PBD-binding peptides suffer from poor potency in cell-based assays, likely due to poor membrane permeability and susceptibility to phosphatase-mediated deactivation. Furthermore, while high affinity ligands for individual Plk1 domains have been reported, the absence of structural data showing the relative orientation of the KD and PBD domains has impeded the design of multi-valent ligands.
The disclosure overcomes these challenges by designing novel high affinity peptide mimetics that incorporate phosphatase stable pThr analogs, specifically Pmab and its C-3 substituted derivatives, leading to improved biochemical potency and cellular uptake. Cyclized iterations of these peptidomimetics yield macrocyclic ligands with enhanced affinity, potency, and pharmacokinetic properties. Moreover, bi-valent ligands incorporating moieties that simultaneously bind to the KD and PBD domains have been developed with effective linkers, resulting in significantly enhanced binding affinity and kinase inhibition, thus representing a potent class of anti-cancer therapeutics.
Claims Coverage
The patent includes four independent claims focusing on bivalent peptido-mimetic ligands, their components, and pharmaceutical compositions thereof. The main inventive features encompass the design of bivalent ligands that bind both kinase and polo-box domains, specific structural features of these ligands, and their pharmaceutical applications.
Structure of bivalent peptido-mimetic ligand
A bivalent peptido-mimetic ligand defined by the structure KD-IDL-PBD, wherein KD is a kinase domain-binding ligand, IDL is a flexible interdomain linker comprising a bond or chemical group, and PBD is a polo-box domain-binding ligand.
Polo-box domain ligand structure
The PBD ligand in the bivalent peptido-mimetic ligand has specified structural features including substituents, linkers, and aryl or heteroaryl groups as defined in the patent, enabling high affinity and selectivity for the Plk1 PBD.
Kinase domain ligand structure
The KD ligand component is defined by specific chemical structures with substitutions including alkyl, cycloalkyl, or aromatic rings, designed to bind the kinase domain selectively and effectively.
Selection of specific bivalent ligand structures
Bivalent peptido-mimetic ligands comprising specific covalent linkages and motifs that optimize simultaneous binding to both KD and PBD, with variations in linker length, attachment points (N- or C-terminal), and incorporation of pharmaceutically acceptable salts, solvates, hydrates, or stereoisomers.
The claims collectively cover the design of bivalent peptido-mimetic ligands that simultaneously target the kinase and polo-box domains of Plk1 via specified flexible linkers, including detailed structural definitions of ligand components and their pharmaceutical compositions, offering potent and selective anti-cancer agents.
Stated Advantages
Enhanced inhibitory potency against Plk1 in biochemical assays through utilization of phosphatase-stable pThr analogs and macrocyclization of peptide mimetics.
Significant enhancement of binding affinity and specificity by designing bi-valent ligands capable of simultaneous binding to kinase and polo-box domains.
Improved pharmacokinetic properties and cellular uptake of peptide derivatives and peptidomimetics due to macrocyclization and structural modifications.
Potential avoidance of off-target effects and cytotoxicity issues seen with ATP-competitive kinase inhibitors by targeting the PBD selectively.
Documented Applications
Use of novel compounds as anticancer therapeutics targeting hyperproliferative disorders such as various human cancers including Acute Lymphoblastic Leukemia, Acute Myeloid Leukemia, Breast Cancer, Colon Cancer, Lung Cancer, Melanoma, Pancreatic Cancer, Prostate Cancer, and others enumerated in the patent.
Methods for prevention, amelioration, or treatment of subjects with hyperproliferative disorders by administering effective amounts of the compounds.
Use in pharmaceutical compositions and kits for delivering anticancer agents specifically inhibiting Plk1 activity through PBD and kinase domain binding.
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