Multisubunit ion channels can adopt pharmacologically distinct forms depending on subunit stoichiometry, accessory factors or post-translational modifications. Production and screening of in vitro correlates that accurately reflected NaV1.7 as it exists in vivo addressed this biological complexity and resulted in compounds that blocked pain in vivo. We used Chromovert® technology to produce numerous cell lines comprising NaV1.7 alpha and beta subunits. Comparisons to alpha-only cell lines demonstrated that minimized target entities do not approximate the intact, multisubunit ion channel. Moreover, cross-comparative pharmacological characterization of numerous multisubunit cell lines enabled selection of a subset that were demonstrated as accurate in vitro correlates of NaV1.7 activity in vivo. At least 20 structurally distinct chemical series identified by parallel HTS of a high-diversity chemical compound library demonstrated efficacy in animal pain models, some with potencies greater than Gabapentin. Efficacious and potent clinical leads with desired selectivity and safety for other complex targets may be developed by coupling previously existing drug development infrastructure and expertise with the new speed and scale with which it is now possible to implement physiologically relevant panels of cell based assays. Results for ENaC and panels of CFTR mutants, additional multisubunit targets and panels that had been out of reach in their intact and native forms, will also be presented.
Presentation: Novel proteolyzed ENaC isoforms and corresponding salt taste enhancing compounds
Modulators of novel proteolyzed human ENaC isoforms were identified and confirmed in sensory testing to either potentiate or block human salt taste perception. First, a stable ENaC cell line was produced using Chromovert(r), a method that enables testing of millions of individual cells to rapidly identify and isolate individual clones stably expressing all desired native subunits. Next, a functional 384-well assay specific for the activity of ENaC comprising alpha, beta and gamma subunits was produced. The cell line was treated with limiting proteolysis in conjunction with the functional assay and a series of novel proteolytic ENaC isoforms was defined based on the differing pharmacology of reference compounds. Interestingly, ENaC isoforms that were less sensitive to inhibition by amiloride were identified, providing a cell based platform consistent with both amiloride-sensitive and amiloride-insensitive components of human salt taste perception. High throughput screening of proteolyzed and non-proteolyzed isoforms resulted in at least 12 distinct chemical series with varied activity against the ENaC isoforms. The creation of and access to multiple distinct ENaC isoforms allowed discovery of corresponding compounds for use as research tools to determine which isoforms correlate with in vivo ENaC activity. Medicinal chemistry and testing of compounds in Ussing chamber models and against delta ENaC is being pursued to further improve nanomolar active compounds for desired safety and efficacy in taste and ENaC-mediated clinical indications including chronic obstructive pulmonary disease (COPD), Cystic Fibrosis and pulmonary edema. Creation of assays for previously inaccessible native and untagged targets comprising all required subunits can aid research in a broad range of applications.
The peace conference this year will be from Sept 19-23 in Jackson Hole, Wyoming. Kambiz Shekdar will be presenting Automated cell engineering: Production of cell based assays that are accurate in vitro correlates of in vivo target activity and faster biologics production on Sunday, September 20th.
The Chromovert drug discovery platform uses transient fluorogenic nucleic acid probes to scan millions of clones for accelerated isolation of rare, optimal cell lines even for targets that had eluded prior reduction to cell based assays. Within three months, cell based assays that represent accurate in vitro correlates of in vivo target activity “IVC” can be produced for any target regardless of its sequence or biology. Data across a diverse set of challenging targets is presented and compared to traditional production methods (e.g., addition of tags or reliance on truncated or modified receptors) and targets lacking all or accessory subunits.
Location: Hunter College of the City University of New York Department of Biological Sciences Fall 2009 Seminar Series
On September 15th, Kambiz Shekdar will be presenting Chromovert Technology: Rapid Cell Engineering for Systems Biology Applications.
For the first time, it is possible to accurately model complex ion channels and receptors as they exist in vivo with the scalability to effectively navigate systems biology applications. Many ion channels and receptors are expressed in specialized differentiated cells (e.g., neurons, epithelial cells) and have been difficult to reliably model in heterologous cell based assays (e.g., CHO, 293). Chromovert technology uses transient fluorogenic nucleic acid probes to scan millions of individual cells for accelerated isolation of rare, optimal cell lines even for heteromultimeric ion channels and receptors that had eluded prior reduction to cell based assays. Within three months, cell based assays that represent accurate in vitro correlates of in vivo target activity can be produced for any target regardless of its sequence or biology.
Title & Topics:
Non-Academic Options for Biology PhDs: Challenging Science and Business Opportunities in Industry.
From widget making to exploiting opportunities of unprecedented scale: the emergence of high caliber
industry science
Starting a company right after grad school
Establishing a creative setting for scientists
Setting and meeting goals: Unique requirements and challenges to securing positions and excelling in industry
Date: Thursday, 06/25/2009 Time: 6:00 PM Location: Robert Wood Johnson Medical School (UMDNJ Campus),
Dean’s Conference Room (Rm-123)
675 Hoes Lane West, Piscataway, NJ 08854
Dr. Kambiz Shekdar received his B.A. from Rutgers University in 1995 and his Ph.D. from the lab of Nobel Laureate Gunter Blobel at The Rockefeller University in 2003. Right after graduate school, Dr. Shekdar co-founded Chromocell Corporation to commercialize the Chromovert technology invented from the lab. The company is focused on several drug discovery programs in pain relief, anxiety treatment and respiratory disorders and also engaged in research and development for Chromovert technology applications in cell therapy, stem cells and other fields.
Please also find the attached flyer for details of this event. Although this is an open invitation and we encourage all to extend the invitation to any interested parties, we ask that ALL attendees please RSVP at the following e-mail address: secretary@rubiotech.org
About IESS: The Industry Expert Seminar Series (IESS) is intended to educate graduate, medical and post-doctoral students in the Rutgers University and UMDNJ community of the opportunities of advanced degree holders within industry, and to expose them to the non-academic career track. It is the hope of the Biotech & Pharma Association at Rutgers and UMDNJ (RUBiotech) that the IESS will not only facilitate awareness of strategies to prepare for positions in industry, but also foster dialogue and networking with current industry leaders.
North Brunswick, NJ –New Jersey Network featured Chromocell’s Intership Program on their Wednesday news section.
Courtesy of NJN
March 25th, 2009
North Brunswick, NJ - New Jersey Network featured Chromocell’s Ribbon Cutting and new research facility on their Friday news section. Courtesy of NJN
March 25th, 2009
North Brunswick, NJ – Ribbon cutting ceremony for new research facility in North Brunswick, NJ [read the press release] Photo by Arthur Paxton
March 19th, 2009
North Brunswick, N.J. – The founders of Chromocell along with the New Jersey Economic Development Authority will be celebrating the opening of the new research facility with a ribbon-cutting ceremony on Wed. March 25, 2009, 2 – 3 pm. For more information, please contact info@chromocell.com.
about chromovert
