Lung, NASH, Scleroderma - Collagen I translation inhibitors
Six compound clusters showing favorable SAR properties are optimized towards different fibrosis indications, Lung and Liver fibrosis and Scleroderma.
Fibrosis represents a large group of diseases that remains without effective treatment. It is caused when a wound healing process goes out of control and progresses into a continuing scarring process in the affected organs. Pulmonary (lung) fibrosis affects around 5 million people around the world. Renal (kidney) fibrosis affects 57 million and liver fibrosis is one of the top ten causes of death worldwide.
The basic fibrotic process, in all types of fibrosis, is related to the over-production of the protein collagen, in response to an injury. The key issue in finding anti-fibrotic drugs is that collagen is not a "target" in the common sense – it is not an enzyme and is not involved in chemical reactions. It exists in huge quantities as it is used to build the body's skin, bones, and connective tissues. Thus, searching for small molecules that will bind to collagen to "neutralize" it is not a valid therapeutic strategy.
During protein translation, ribosomes chain amino acids that are carried by tRNAs molecules. Anima’s Protein Synthesis Monitoring (PSM) technology uses fluorescently labelled tRNA pairs, specifically selected for each target protein. When such a labeled pair enters the ribosome, it emits a light pulse. The light pulses tell us when, where and how much of the target proteins being made by ribosomes, in real time.
We follow Collagen I translation by labeling a the tRNA pair tRNApro, tRNAgly. This pair repeats in high frequency in Collagen I mRNA and is therefore its “signature” pair.
The images below show Anima's technology, PSM, being used to detect the collagen’s I translation. The dots of light represent ribosomes actively translating Collagen Type I in activated fibroblasts. Using siRNA specific to Collagen I (middle and right images), less light is detected, indicating a reduction in Collagen I translation. In our screens, hit compounds reduce the light as they decrease the production of collagen.
Our platform works in high content screening, 384-well plate format. A diverse library of 100,000 compounds was screened, generating a set of 20 million images. The big data is then analyzed by our cloud based software using proprietary image analysis and machine learning algorithms identifying “hits”, compounds that are actively inhibiting the production of Collagen.
Discovered Hit compounds were shown to be specific to Collagen I translation, not inhibiting general protein translation. As a further validation, we showed with Immunofluorescence against collagen that hits reduce collagen translation, but do not completely inhibit it. This indicates that the compounds only affect the induction of collagen expression, but not steady state levels.
Hits were shown to be active in a dose dependent manner in down regulating collagen I translation