Patents
1. Protein controlling synthesis of collagen and associated methods.
Inventor: Branko Stefanovic
U.S. Patent Application Serial No. 13/706,747
ADDMG Ref: 0112890_CON
FSU Ref: 10-042 Stefanovic
Invention. This invention relates to the biomolecular mechanism controlling synthesis of type I collagen, which is overproduced in various forms of fibrosis. The invention advantageously discloses the previously unknown protein with binding affinity for the 5' stem loop in collagen mRNAs. The present disclosure is the first to identify this protein, LARP6, as responsible for activating translation of collagen mRNAs. The invention discloses an isolated purified polypeptide having an amino acid sequence of LARP6 and having binding affinity for a 5' stem-loop in mRNAs encoding type I collagen. The polypeptide disclosed herein may be employed in a method of screening an agent for ability to interfere with collagen synthesis. The method comprises reacting the disclosed polypeptide with collagen mRNAs in presence of the agent, and detecting if the agent has interfered with binding of the polypeptide to the mRNAs.
Significance. The discovery that LARP6 regulates production of type I collagen in fibrosis opened a possibility that inhibitors of LARP6 binding to collagen mRNAs can be specific antifibrotic drugs. This is an attractive prospect for entities involved in drug discovery, such as pharmaceutical companies, research institutes and nonprofit organizations, especially because fibrosis is very common and there are no approved antifibrotic drugs. This patent protects the intellectual property of using LARP6 as the target for drug discovery.
2. Compositions and methods for inhibiting collagen production
Inventor: Branko Stefanovic
U.S. Patent Application Serial No. 13/646,001
ADDMG Ref: 0116975_UT
FSU Ref: 12-047 Stefanovic
Invention. The inventors have developed small molecule inhibitors of excessive type I collagen production in fibrosis. The inventors used a fluorescence-polarization technique to develop an assay for screening small molecules that could potentially inhibit the binding of LARP6 and the stem loop. The inventors described a few compounds that they had identified using this assay from a library of ~50,000 that had inhibitory effect on type I collagen. Several chemical compounds were identified in the screen and tested for their ability to inhibit type I collagen production by cells in culture. The compound 60D17 (also termed C9) strongly inhibited type I collagen.
Validation of 60D17 in animal models of hepatic fibrosis. The validation of antifibrotic activity of the compound 60D17 came from 3 animal studies. In the first therapeutic study bile duct ligation was used to model hepatic fibrosis. When left untreated, the fibrosis progressed until day 10 after the ligation, however, when treated with 1 mg/kg of 60d17 from day 2.5 to day 10, the progression of fibrosis was inhibited by >50%.
The second therapeutic model was ethanol/CCl4 induced liver fibrosis. After 3 weeks of induction a moderate fibrosis developed, which was treated with 1 mg/kg of 60D17. In rats receiving vehicle during the treatment period the fibrosis dramatically progressed. In 60D17 treated rats the progression of fibrosis was significantly attenuated.
In a prophylactic CCl4 model the induction of fibrosis and the treatment was at the same time. After 4 weeks an advanced fibrosis was developed in rats receiving vehicle but in animals receiving 60D17 the development of fibrosis was completely prevented.
Licensing of 60D17. The 60D17 compound was licensed to a start up company. The goal is to develop the first in class antifibrotic drug based on the structure of 60D17, which would be effective for treatment of all forms of fibrosis.
3. Characterization of type 1 collagen synthesis by fluorescence imaging and related drug screening
technique.
Inventor: Branko Stefanovic
EFS ID: 31382336
Confirmation Number: 3257
International Application Number: PCT/US18/12045
Invention. To develop the collagen biosensor chimeric genes were constructed including an mTagBFP2 sequence incorporated within the sequence of human collagen α1(I) polypeptide (COLα1(I)/mTagBFP2, blue fluorescence) and mEmerald sequence incorporated within the human collagen α2(I) polypeptide (COLα2(I)/mEmerald, green fluorescence). The chimeras were constructed in such a way that the sequences of the fluorescent proteins do not substantially interfere with normal synthesis, modifications and assembly of the collagen polypeptides. The visualization of the individual collagen polypeptides is achieved by fluorescence imaging in blue and green channels and their co-localization by double fluorescence imaging.
Significance. The technology visualizes synthesis of type I collagen in live cells. It demonstrated that the synthesis is organized in discrete granules, collagenosomes, which can be quantified. By scoring for collagenosome formation, chemical compounds active in blocking excessive type I collagen formation can be discovered. The technology is suitable for high throughput screening of chemical libraries.