Mechanism and Uses of Transmembrane Helix Insertion by Soluble Peptides

可溶性肽跨膜螺旋插入的机制和用途

• 批准号: 10089455
• 负责人: Oleg A Andreev
• 金额: $ 57.93万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10089455
• 关键词: Acidity; Algorithms; Amanitins; Antibodies; Antibody-drug conjugates; Area; Basic Science; Bioinformatics; Biological Markers; Breast Cancer Patient; C-terminal; Cell surface; Cells; Chemicals; Clinical Trials; Computer Models; Cyclic Peptides; DNA; Data Set; Databases; Development; Diagnosis; Disease; Excipients; Fluorescent Dyes; Goals; Grant; Hand; Hydrophobicity; Kinetics; Learning; Length; Link; Lipids; Malignant neoplasm of urinary bladder; Measures; Mediating; Medicine; Membrane; Memorial Sloan-Kettering Cancer Center; Methods; Modeling; Modernization; Molecular; Operative Surgical Procedures; Outcome; Peptides; Perfusion; Periodicity; Permeability; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Positron-Emission Tomography; Property; Publishing; RNA Polymerase II; RNA Polymerase Inhibitor; Research; Resistance; Solubility; Structure; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Therapeutic Studies; Toxin; Tubulin; Tumor Markers; Variant; Vesicle; Work; alpha helix; base; biophysical techniques; cancer cell; cancer imaging; clinical application; design; expectation; fluorescence imaging; fluorescence-guided surgery; image guided; imaging agent; in vivo; insight; iterative design; model design; molecular dynamics; neoplastic cell; non-drug; novel; novel diagnostics; nuclear imaging; pH gradient; polypeptide; side effect; targeted agent; targeted delivery; targeted imaging; targeted treatment; treatment strategy; tumor

PROJECT SUMMARY/ABSTRACT    The  understanding  we  now  have  of  the  pHLIPs  (pH-­Low  Insertion  Peptides)  enables  the  design  of  novel,  pH-­ sensitive  targeting  agents  that  use  the  acidity  of  tumor  cell  surfaces  as  a  biomarker.  The  work  supported  by  this  grant  has  already  yielded  a  new  diagnostic  nuclear  imaging  agent  (PET-­pHLIP)  and  a  new  fluorescent  imaging  agent  for  image-­guided  surgical  interventions  (ICG-­pHLIP),  which  are  advancing  to  clinical  trials  at  Memorial Sloan Kettering Cancer Center in 2019.   The  primary  focus  of  this  continuation  is  to  enable  targeted  intracellular  delivery  of  polar  and  moderately  hydrophobic  therapeutic  molecules.  We  propose  a  systematic  approach  using  representative  cargoes  from  3  classes  of  therapeutic  molecules  that  possess  different  physical,  chemical  and  functional  properties:  i)  a  moderately  hydrophobic,  sparingly  cell-­permeable,  small  drug  molecule:  mertansine;;  ii)  a  moderately  polar, cell-­impermeable,  cyclic,  rigid,  larger  drug:  amanitin;;  and  iii)  a  large,  polar,  cell-­impermeable  drug:     calicheamicin.  Each  of  these  drugs  is  currently  under  development  or  in  use  as  an  antibody-­drug  conjugate  warhead,  but  there  are  important  limitations  to  the  antibody  approaches,  including  limited  biomarker  availability,  resistance  selection,  a  narrow  therapeutic  window  and  limited  delivery,  often  with  <  1%  of  a  construct reaching the tumor. Our approach is based on targeting tumor acidity, especially cancer cell surface  pH that is a general parameter within and among different tumors, and independent of tumor perfusion.   We  propose  to  explore  variation  of  pHLIP  sequences,  introduce  pHLIP-­cycles  and  exploit  pHLIP-­bundles  for  delivery  of  these  therapeutic  cargoes.  The  pHLIP-­cargo  constructs  will  be  tested  for  insertion  stability  and  kinetics,  using  biophysical  methods  and  computational  modelling.  Activity  will  be  evaluated  in  cells,  and  promising  constructs  will  be  assessed  in  vivo.  We  will  accumulate  a  parameter  database  of  pHLIP-­cargo  properties  and  employ  modern  bioinformatics  algorithms  to  analyze  the  entire  data  set  to  reveal  major  design  principles.  By studies of these therapeutic agents with pHLIPs, we hope to find principles for targeted delivery of a range  of  other  compounds.  Should  we  be  successful,  the  limitations  of  antibody  drug  conjugates  for  therapy  will  be  overcome, and greater understanding of the membrane barrier will also be in hand.  Our expectation is to have  a candidate for the treatment of bladder cancer as the practical outcome of this grant renewal.

项目总结/文摘