Recombinant adenoviral vectors and their utility in the treatment of various types of fibrosis: hepatic, renal, pulmonary, as well as hypertrophic scars

Abstract

SUMMARY OF THE INVENTION The use of gene therapy for the treatment of different kinds of fibrosis in human beings is disclosed. The purpose is the use of “therapeutic2 genes specifically directed to target organs to revert and/or prevent the development of the fibrosis process. The potential application of gene therapy to patients with fibrosis and/or cirrhosis will depend to a large extent on the successful delivery of genes which encode for therapeutic proteins to livers with severe fibrosis and that these genes which encode for proteins human MMP-8 active and latent, MMP-1, MMP-2, MMP-9 and MMP-13; human uPA wild type and/or modified (or its truncated version), the truncated receptor for TGF-β type II and Smad-7 can be directed by adenovirus and/or other recombinant vectors that cannot transduce (infect) others organs. The recombinant adenoviruses (AdR) are vectors highly efficient for the transduction of therapeutic genes to diverse target cells. We have proved that they can carry genes to cirrhotic livers. The delivery of therapeutic genes through such adenoviral vectors and other recombinant vectors could also be performed using cationic and anionic liposomes (DOTMA). Therefore, we propose the use of this patent to be applied in the same manner to: Renal fibrosis Pulmonary fibrosis Hypertrophic and keloid scars (skin fibrosis), and Other kinds of fibrosis.

1 .- A recombinant adenoviral vector which contains an adenoviral genome from which the open reading frames E 1  and/or E 3  have been deleted, but retains enough sequence to make the adenoviral vector able to replicate in vitro, said vector also contains a therapeutic gene or a DNA sequence of interest regulated by ubiquitous promoters and/or tissue-specific promoters that encodes for therapeutic proteins useful in fibrosis treatment of the fibrosis.  
     
     
         2 .- The recombinant adenoviral vector according to  claim 1 , in which the specific tissue-promoter is PEPCK.  
     
     
         3 .- The recombinant adenoviral vector according to  claim 1 , in which the therapeutic gene or the DNA sequence cloned in such adenoviral vector is selected from latent and active human metalloprotease gene MMP-8, MMP-1, MMP-2, MMP-9 and MMP-13; ; human urokinase Plasminogen Activator gene (uPA wild type and/or modified), gene of the truncated receptor for TGF-β type II; and Smad 7 which encode for therapeutic proteins, that degrade excess of collagenic proteins deposited in the cirrhotic organs.  
     
     
         4 .- The recombinant adenoviral vector according to  claim 3 , in which the therapeutic gene is a DNA sequence selected from the gene of the Hepatocyte Growth Factor (HGF), which encodes for proteins stimulators of hepatic regeneration with the purpose to re-establish the normal functions of the liver.  
     
     
         5 .- The recombinant adenoviral vector according to  claim 1 , in which the therapeutic proteins for the treatment of fibrosis are the latent and/or active protein MMP-8, MMP-1, MMP-2, MMP-9 and MMP-13; uPA wild type and/or modified; the truncated receptor for TGF-β type II; betaglycan; HGF and Smad 7.  
     
     
         6 .- The recombinant adenoviral vector according to  claim 1 , which contains also the delivery of therapeutic genes or DNA sequences which encode for therapeutic proteins intended for the treatment of fibrosis in cirrhotic liver.  
     
     
         7 .- The recombinant adenoviral vector according to  claim 6 , in which the delivery of the therapeutic genes is carried out in other organs with generalized fibrosis.  
     
     
         8 .- The recombinant adenoviral vector according to  claim 7 , in which the tissue-specific recognition of the therapeutic genes to the organs with fibrosis, is conducted by the administration route used.  
     
     
         9 .- The recombinant adenoviral vector according to  claim 8 , in which the administration route is endovenous.  
     
     
         10 .- The recombinant adenoviral vector according to  claim 6 , in which the organs with fibrosis are selected from liver, lung, heart, kidney, skin, and hypertrophic scars.  
     
     
         11 .- The recombinant adenoviral vector according to  claim 10 , in which the main target organ is the liver.  
     
     
         12 .- Recombinant adenoviral vectors according to  claims 1  to  11  in which the delivery of therapeutic genes is realised through the use of viral or non viral vectors.  
     
     
         13 .- The recombinant adenoviral vector according to  claim 12 , in which non viral vectors are selected from plasmids and cationic and anionic liposomes.  
     
     
         14 .- The recombinant adenoviral vector according to  claim 6 , in which the efficient sending of collagenase gene MMP-8 to cirrhotic liver, can induce the degradation of collagen by means of over-expression of metalloproteases.  
     
     
         15 .- The recombinant adenoviral vector according to  claim 1 , characterized because it is used for the treatment of the hepatic, pulmonary, renal, heart fibrosis, keloids and hypertrophic scars, and which does not induce lethal toxicity.  
     
     
         16 .- A process to prepare recombinant adenoviral vectors through the cloning of reporter genes Lac-Z and GFP and the therapeutic gene, which encodes for therapeutic proteins for the treatment of hepatic, pulmonary, renal, and/or heart fibrosis, keloids and hypertrophic scars.  
     
     
         17 .- The process according to  claim 16  in which the therapeutic gene is selected from the human metalloprotease gene MMP-8 latent and active, MMP-1, MMP-2, MMP-9 and MMP-13; gene for human uPA wild type and/or modified; Smad 7 and gene of the truncated receptor of TGF-β type II.  
     
     
         18 .- The process according with  claim 16  in which the recombinant adenoviral vector is pAdGFP-MMP-8.  
     
     
         19 .- A pharmaceutical composition containing a therapeutically effective amount with a regimen of unitary doses of viral particles of recombinant adenoviral vectors, according to  claim 1 , for the treatment of hepatic, pulmonary, renal, and heart fibrosis, keloids and hypertrophic scars, combined with a pharmaceutically compatible carrier.  
     
     
         20 .- The pharmaceutical composition according to  claim 19 , in which the unitary dose is of about 10 7 -10 14  viral particles for an individual with fibrosis.  
     
     
         21 .- The use of recombinant adenoviral vector according with  claim 1 , for the elaboration of a bio-medication for the treatment of hepatic, pulmonary, renal, and heart fibrosis, keloids and hypertrophic scars.