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Design, Synthesis, and Biological Activity of Rhodium Metalloinsertors

Citation

Komor, Alexis Christine (2014) Design, Synthesis, and Biological Activity of Rhodium Metalloinsertors. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/KPCY-JS09. https://resolver.caltech.edu/CaltechTHESIS:05282014-143107089

Abstract

Deficiencies in the mismatch repair (MMR) pathway are associated with several types of cancers, as well as resistance to commonly used chemotherapeutics. Rhodium metalloinsertors have been found to bind DNA mismatches with high affinity and specificity in vitro, and also exhibit cell-selective cytotoxicity, targeting MMR-deficient cells over MMR-proficient cells.

Here we examine the biological fate of rhodium metalloinsertors bearing dipyridylamine ancillary ligands. These complexes are shown to exhibit accelerated cellular uptake which permits the observation of various cellular responses, including disruption of the cell cycle and induction of necrosis, which occur preferentially in the MMR-deficient cell line. These cellular responses provide insight into the mechanisms underlying the selective activity of this novel class of targeted anti-cancer agents.

In addition, ten distinct metalloinsertors with varying lipophilicities are synthesized and their mismatch binding affinities and biological activities studied. While they are found to have similar binding affinities, their cell-selective antiproliferative and cytotoxic activities vary significantly. Inductively coupled plasma mass spectrometry (ICP-MS) experiments show that all of these metalloinsertors localize in the nucleus at sufficient concentrations for binding to DNA mismatches. Furthermore, metalloinsertors with high rhodium localization in the mitochondria show toxicity that is not selective for MMR-deficient cells. This work supports the notion that specific targeting of the metalloinsertors to nuclear DNA gives rise to their cytotoxic and antiproliferative activities that are selective for cells deficient in MMR.

To explore further the basis of the unique selectivity of the metlloinsertors in targeting MMR-deficient cells, experiments were conducted using engineered NCI-H23 lung adenocarcinoma cells that contain a doxycycline-inducible shRNA which suppresses the expression of the MMR gene MLH1. Here we use this new cell line to further validate rhodium metalloinsertors as compounds capable of differentially inhibiting the proliferation of MMR-deficient cancer cells over isogenic MMR-proficient cells. General DNA damaging agents, such as cisplatin and etoposide, in contrast, are less effective in the induced cell line defective in MMR.

Finally, we describe a new subclass of metalloinsertors with enhanced potency and selectivity, in which the complexes show Rh-O coordination. In particular, it has been found that both Δ and Λ enantiomers of [Rh(chrysi)(phen)(DPE)]2+ bind to DNA with similar affinities, suggesting a possible different binding conformation than previous metalloinsertors. Remarkably, all members of this new family of compounds have significantly increased potency in a range of cellular assays; indeed, all are more potent than the FDA-approved anticancer drugs cisplatin and MNNG. Moreover, these activities are coupled with high levels of selectivity for MMR-deficient cells.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:DNA, chemotherapeutic, mismatch, metalloinsertion
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Chemistry
Awards:The Herbert Newby McCoy Award, 2014
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Barton, Jacqueline K.
Thesis Committee:
  • Gray, Harry B. (chair)
  • Barton, Jacqueline K.
  • Peters, Jonas C.
  • Rees, Douglas C.
Defense Date:23 May 2014
Non-Caltech Author Email:ackomor (AT) gmail.com
Record Number:CaltechTHESIS:05282014-143107089
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:05282014-143107089
DOI:10.7907/KPCY-JS09
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1080/02603594.2014.890099DOICh. 1: The Path for Metal Complexes to a DNA Target
http://dx.doi.org/10.1021/bi2015822DOICh. 2: Selective Cytotoxicity of Rhodium Metalloinsertors in Mismatch Repair-Deficient Cells
http://dx.doi.org/10.1021/ja3090687DOICh. 3: Cell-Selective Biological Activity of Rhodium Metalloinsertors Correlates with Subcellular Localization
http://dx.doi.org/10.1371/journal.pone.0078726DOICh. 4: An Inducible, Isogenic Cancer Cell Line System for Targeting the State of Mismatch Repair Deficiency
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8412
Collection:CaltechTHESIS
Deposited By: Alexis Komor
Deposited On:29 May 2014 21:46
Last Modified:04 Oct 2019 00:05

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