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

Abstract

<heading lvl="1">SUMMARY OF THE INVENTION < / heading> 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-beta 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.

Description

[0001] The present invention relates to the creation of RECOMBINANT ADENOVIRAL vectors bearing exogenous genes that encode for therapeutic proteins useful in the treatment of HEPATIC cirrhosis and generalized FIBROSIS, such as renal FIBROSIS, pulmonary FIBROSIS, HYPERTROPHIC scars and keloid of the skin, and / or in other target organs susceptible to suffer from it. It also relates to a mechanism of tissue-specific recognition of the affected cells by means of delivery of therapeutic genes to cirrhotic organs.[0002] Moreover, the invention provides an effective way for the treatment of fibrosis through the employment of recombinant adenoviral vectors which are claimed here, as well as the process to prepare these vectors, the pharmaceutical composition that contains them, and their therapeutic uses in the treatment of several fibrosis, which has great commercial expectancy in the pharmaceutical industry and also presents an important alternative as gene therapy for the treatment of ch...

AI Technical Summary

Benefits of technology

[0037] Another advantage resulting from the fibrosis treatment is that recombinant adenovirus does not induce lethal toxicity in none of the injected animals with the vectors.

[0038] Another objective of the invention allows us to discriminate the modification of the staining reaction with X-Gal between the endogenous tissue galactosidase activity and the bacterial -galactosidase induced by the infectious action of the adenoviral vector. The use of the green fluorescent protein permits us to verify the in vivo transduction of different organs in rats to verify if the vector administration was appropriate, if the expression remains, and besides not killing the animals it is possible to conduct follow up observation after surgery.

Problems solved by technology

Regardless of the ethiological agent and morphologic differences, all forms of cirrhosis have as a common end, hepatic failure causing the patient's death.

Thus, the liver is not able to maintain the physiologic concentration of solutes in the terminal hepatic vein, in other words, HEPATIC failure sets in.

With an excessive deposition of this protein, the free exchange of nutrients between blood and liver cells is impeded, the inactivation of toxic agents by this organ can not be carried out, becoming this the main cause of the pathophysiology of the disease.

To date, no therapeutic agent that could revert and / or prevent with a 100% effectiveness the progressive accumulation of hepatic collagen has been described.

However, precautions have to be taken since these vectors can generate potential replication-competent viruses.

On the other hand, up to date, no study has reported incidences of mutagenesis by insertion or activation of oncogenes by the incorporation of the replication-deficient retrovirus.

Nevertheless, the use of retroviral vectors to transduce genes to the liver is limited for the following considerations: 1) these vectors infect only cells which actively divide and 2) very low viral particles titers are obtained in the packing cell lines used to amplify these viruses (Graham F L, and Van Der Eb A J. A New Technique for the Assay of Infectivity of Human Adenovirus 5 DNA.

Hepatic cirrhosis is a chronic illness of the liver, where diffuse cell necrosis and a limited regeneration of parenchymal hepatic cells result in diffuse percentage increase of connective tissue, causing the distortion of lobular hepatic architecture and inducing hemodynamic alterations.

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