Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

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

a technology of adenoviral vectors and fibrosis, which is applied in the direction of plant growth regulators, enzymes, biocide, etc., can solve the problems of patient's death, liver not being able to maintain the physiologic concentration of solutes, and hepatic failur

Inactive Publication Date: 2003-01-02
TGT LAB DE C V
View PDF18 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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.

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.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Recombinant adenoviral vectors and their utility in the treatment of various types of fibrosis: hepatic, renal, pulmonary, as well as hypertrophic scars
  • Recombinant adenoviral vectors and their utility in the treatment of various types of fibrosis: hepatic, renal, pulmonary, as well as hypertrophic scars
  • Recombinant adenoviral vectors and their utility in the treatment of various types of fibrosis: hepatic, renal, pulmonary, as well as hypertrophic scars

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0073] Methodology to demonstrate the activity of Metalloprotease or Collagenase (MMP-8) and how to regulate its function

[0074] a) Cell Culture

[0075] HepG2 cells is a cell line of parenchymal origin derived from a human hepatoma, and were cultured in 60 mm culture dish, 37.degree. C. in a wet atmosphere, with 95% air and CO.sub.2 5% atmosfere in Eagle's medium modified by Dulbecco (DMEM), supplemented with 10% fetal bovine serum, 2 mM L-Glutamax and antibiotics (100 U / ml penicillin and 100 g. / ml. streptomycin).

[0076] b) Vectors of Expression of Latent and Active MMP-8 Genes

[0077] Two plasmids were used with 2 kinds of MMP-8 genes to transfect the hepatic cells: The plasmid pcDNA-MMP-8 which contains the cDNA which encodes for latent MMP-8 (pro-MMP-8) together with the strong viral promoter of cytomegalovirus (CMV); and the plasmid pcDNA.sub.3MMP-8 containing the cDNA which encodes for the active MMP-8, together with the CMV promoter. This last one was created through subclonation us...

example 2

[0098] Results to Demostrate the Activity of Metalloprotease or Collagenase (MMP-8) and therefore to Regulate its Function

[0099] Subcloning permitted to incorporate MMP-8 cDNA encoding for the fully functional enzyme was subcloned in a vector appropriate to our needs. Thus, FIG. 15 shows an electrophoresis of the DNA fragments released by cutting MMP-8 plasmids with restriction enzymes. Lane A). Marker of bp of 1 Kb DNA ladder (Gibco BRL); B). Perfect DNA marker (Novagen, Inc.); 1) pcDNA-MMP-8 cutting with BamHI and XbaI; 2) pcDNA3-MMP-8 cutting with BamHI and XbaI; C) .phi.X174 marker (Gibco BRL); .lambda. Hind III Marker (Gibco BRL), in which the latent MMP-8 cDNA (lane 1) and the mature MMP (lane 2); were successfully subcloned in the expression vectors pcDNA and pcDNA3. The released inserts are observed after treatment with restriction enzymes BamH1 and XbaI. The bands stained with ethydium bromide correspond to each of cDNA (between 506 and 560 base pairs) for mature and latent...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
pHaaaaaaaaaa
pHaaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

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...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/76A61K35/761A61K38/46C12N15/09A61K48/00A61P1/16A61P9/00A61P11/00A61P13/12A61P17/02A61P37/02A61P43/00C12N9/64C12N15/861
CPCA61K48/00C12N7/00C12N9/6491C12N15/86C12N2710/10243C12N2830/008A61P1/16A61P11/00A61P13/12A61P17/02A61P37/02A61P43/00A61P9/00
Inventor ARMENDARIZ BORUNDA, JUANAGUILAR CORDOVA, ESTUARDO
Owner TGT LAB DE C V
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products