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SAN DIEGO, CA, United States

Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 173.02K | Year: 2009

DESCRIPTION (provided by applicant): Ion channel targeted drug discovery has been hampered by the unavailability of cell-based assays that utilize physiological and pharmacological relevant measurements to characterize compound activity. This project will develop an assay to measure intracellular chloride ion and pH as direct physiological measurements to interrogate the chloride ion channels and proton transporter(s) for GI drug discovery. The Phase I of this proposal will focus on the synthesis, characterization and validation of the chloride ion sensing indicator using T84 cells. The Phase II of this proposal will focus on the development of the pH sensing indicator, integration of the pH sensing assay with the Cl assay; validation of the joint chloride and pH assay using T84 cells by delineating the intracellular coupling dynamics of [Cl-]i and pHi for GI drug discovery; and scaling up the manufacturing of the assays for commercialization. PUBLIC HEALTH RELEVANCE: Ion channels of chloride and proton transports are the two predominant drug discovery gastrointestinal (GI) target sites for the treatments of diarrhea, constipation and gastric reflux. Cytoptics Corporation proposes to develop a new generation of chloride ion and pH sensing indicators to measure intracellular chloride ion concentrations, [Cl-]i, and pHi simultaneously for cell-based GI drug discovery. This Phase I proposal will focus on the 1) synthesis and spectral characterization of chloride ion sensing indicator; 2) development of the loading protocols of the indicator; and 3) physio-pharmacological measurements of [Cl-]i in T84 cells. This will enable a chloride ion sensing assay kit to be readily marketed for GI drug discovery and development following the completion of the Phase I. The Phase II of this proposal will focus on the development of the pH sensing indicator, integration of the pH sensing assay with the Cl assay; validation of the joint chloride and pH assay using T84 cells by delineating the intracellular coupling dynamics of [Cl-]i and pHi for GI drug discovery; and scaling up the manufacturing of the assays for commercialization.


Cytoptics Corporation | Entity website

Cell-Based Fluorescent Microscopy & HTS Cytoptics provides to industry, academia and government, the evaluation of targeted agents using novel technologies and cell lines for ionic mechanism-dependent drug discovery, validation and safety pharmacology. Please send inquiry ...


Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 1.03M | Year: 2011

DESCRIPTION (provided by applicant): Ion channel drug discovery has been hampered by the lack of cell-based high throughput screening (HTS) assay to physiologically and pharmacologically characterize compound activities. Cytoptics Corporation (Cytoptics) proposes to develop a new generation of chloride ion and pH sensing luminous quantum dots (QD) indicators to measure intracellular chloride ion concentrations, [Cl-]i, and pHi for fluorescence microscopy and cell-based high throughput screening (HTS) targeting GI drug discovery. These fluorescence microscopy and cell-based HTS assays utilize the size-tunable, non- overlapping emission wavelengths, ion-selective QD to measure [Cl-]i and pHi simultaneously in T84WT /T84C1C2 as well as in HEK293WT/HEK293C1C-2 cells. Using the quantum dots (QD) as Forester Resonance Energy Transfer (FRET) donors and the respective Cl- and H+ chemical sensors conjugated to the QD as acceptors, these two nanosensors, Cl-QD and pH-QD, quench their fluorescence intensities of the QDaccording to the concentrations of the Cl- and H+ bound to their respective Cl- and H+ sensors. We have accomplished all the proposed Phase I objectives, namely, 1) synthesis and spectral characterization of chloride ion sensing luminous QD, Cl-QD; 2) development of a loading protocol of the Cl-QD into T84 cells; and 3) measurements of the physio-pharmacological [Cl-]i responses in T84WT and T84C1C-2 cells using the Cl- QDTM. In Phase II, Cytoptics will incorporate and extend the accomplishments of Phase Ito produce six assays to target the cell-based GI drug target discovery market. They are (1) the Cl-QDTM assay for fluorescence microscopy; (2) the Cl-QDTM assay for cell- based HTS; (3) the pH-QD assay for fluorescence microscopy; (4) the pH-QD assay forcell-based HTS; (5) the Cl-QDTM and pH-QD integrated assay for cell-based HTS; and (6) the Cl-QDTM and pH-QD integrated assay targeting C1C-2 channels using the stable T84C1C-2 and HEK293C1C-2 cells for GI drug discovery. These novel assays address the critical needs of cell-based HTS drug discovery and will also broadly impact drug discovery research in ionic signal transduction. PUBLIC HEALTH RELEVANCE: Chloride channels and proton transporters are the two predominant drug discovery gastrointestinal (GI) target sites for the treatments of diarrhea, constipation and gastric reflux. Cytoptics Corporation proposes to develop a new generation of chloride ion and pH sensing indicators (Cl-QD and pH-QD, respectively) to simultaneously measure the dynamicsof the intracellular chloride ion concentrations ([Cl-]i) and pH (pHi) in epithelial cells for the first time. These nanosensors will be utilized to interrogate the Cl-channels and H+-transporters as direct physiological and pharmacological assays for GIdrug discovery. Using the quantum dots (QD) with different emission wavelengths for these two nanosensors, Cl-QD and pH-QD emit separate colors with their non-interfering fluorescence intensities varied according to their respective concentrations of Cl- and H+, Cytoptics will produce six assays utilizing these two novel QD-based nanosensors to target the cell-based GI HTS drug discovery market. These novel assays will also provide unparallel insights in the ionic cellular signal transduction mechanisms that heretofore not been possible.


Cytoptics Corporation | Entity website

About us: Cytoptics was founded by Drs. Lid B ...


Cytoptics Corporation | Entity website

Quantum-Dots Nanobiosensors for Fluorescent Microscopy & Cell-Based HTS 1) Cl-QD (TM) -- The chloride indicator, Cl-QD, is a new generation of chloride indicator based on quantum dots technology. It is highly quantum efficient ...

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