Human Cell Based High-throughput Genotoxicity Screen

Anthem-GenTox

In the past year

• It is an engineered human cell line based screen that is compatible for high-throughput analysis. • The same cell line reports four parameters – cytotoxicity, and three genotoxicity read-outs. • The amount of compound required is minimal ~ 10 mg/compound • The assay can be completed and analyzed in 2 days, versus weeks for other conventional assays • Extensive validations have been conducted in-house, at external sites and in collaboration with labs in Europe (the data has been published *). • The sensitivity and specificity values far exceed any of the standard assays currently in use. • It is the best in class assay with respect to concordance with in vivo data.

There have been various assays developed to quantify genotoxicity. They include methods such as a.) bacterial AMES test that depends on reversion of mutations in response to genotoxic agents and counting the colonies that survive on special media, b) manually counting chromosomal aberration in mammalian cells, and c.) manually counting micronuclei in mammalian cells post DNA damage. The methods either rely on DNA damage response mechanisms in bacteria that is very different from those in mammalian/human cells, or tedious and error-prone manual counting of DNA damage events. These methods are also not adaptable to high throughput, generally takes longer, and is more expensive. In response to the many limitations discussed above, our competitor and Anthem came up with an engineered human cell line that would sense and report DNA damage, and thus accurately quantify genotoxicity. Our product differs from the competitor in three ways: a.) there are three DNA damage sensors in our system versus the competitor single sensor, b.) our DNA damage sensors are different from the competitor’s sensor (we use GADD153, p21 and p53 binding elements versus only GADD45a used by our competitor), and c.) our assays provide information on whether or not the compound effects the major DNA damage pathway (the p53 pathway). Our data indicates that the three reporter engineered cell line developed by Anthem is more concordant with animal study data than that of our competitor.

Pharmaceutical industry Chemical Industry Food and agrochemical industry Personal Care Research and Academia Environmental Protection Agency

- Traditional methods are time consuming (weeks) versus the Anthem GenTox is rapid (24-48 hour read-outs) - Traditional methods are prone to human error whereas the Anthem GenTox reporter based system is an enzymatic assay. As a result, it is more precise. - Traditional methods are low throughput. The Anthem GenTox assay is adaptable to high throughput.

- Anthem Biosciences is the inventor of this technology platform. - They employ over 300 scientists covering biology and chemistry - Anthem is a full service provider with offerings ranging from research to manufacturing to animal studies. - Their facility is cGMP and FDA inspected. Together with Anthem, DavosPharma is a marketing partner providing the following expertise. - DavosPharma is a leading provider of discovery services and custom cGMP manufacturing of APIs, NCEs and Biologics through carefully vetted global partners - Over 40 years of integrating drug discovery R&D and Commercialization - Over 300 years of collective Pharmaceutical experience - Paid on deliverables NOT “best efforts”

Pharmaceutical industry Chemical Industry Food and agrochemical industry Personal Care Research and Academia Environmental Protection Agency

Pricing varies on agreement. If screening is done on a small number of compounds, price ranges between $3K-$4K per compound. Otherwise, the technology can be licensed for in-house screening.

This innovation is a novel application (genotoxicity sensing assay) of a known DNA damage pathway.