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Treatment of eye disorders characterized by an elevated intraocular pressure by sirnas

A technology for eye diseases and uses, applied in DNA/RNA fragments, recombinant DNA technology, sensory diseases, etc., can solve problems such as accurate monitoring and prognosis

Inactive Publication Date: 2007-11-21
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0033] At present our existing treatments certainly fall short and the prognosis is plagued by a combination of practical difficulties related to the evaluation of outflow potential, precise monitoring of treatment and complexity of surgical technique; the dominant factor in all glaucomas is the retinal ganglion Degeneration of cells and thus neuroprotection by effective hypotension is a basic requirement for any treatment we use (for a recent review on this issue, see Khaw et al 2004b)

Method used

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  • Treatment of eye disorders characterized by an elevated intraocular pressure by sirnas
  • Treatment of eye disorders characterized by an elevated intraocular pressure by sirnas
  • Treatment of eye disorders characterized by an elevated intraocular pressure by sirnas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Example 1 In Vitro Assay.

[0131] To determine the inhibition of different targets involved in glaucoma using RNAi technology, the first step is to perform experiments in cell culture. For each target, several siRNAs were designed using specific software according to the aforementioned rules. The siRNAs were applied to cell cultures such as NPE, OMDC and HEK293. The effect of siRNAs on the target gene was analyzed by real-time PCR and semi-quantitative PCR according to standard protocols. Transcript levels of the gene targets were normalized using actin as a housekeeping gene. Table I below shows representative results of real-time PCR experiments for some of the above-mentioned target genes. The values ​​represent the mean and their standard deviation of the percentage of siRNA interference on each gene expression that had been normalized to control cells. Different transcript levels at 24 and 48h time points were all significantly reduced after siRNA treatment co...

Embodiment 2

[0149] Example 2 In vivo assay.

[0150] Prior to siRNA therapeutic application, in vivo assays were validated to determine correct siRNA delivery.

[0151] Those siRNAs selected by in vitro assays were applied to animal models, following the procedure described above. To avoid the effect of IOP fluctuations due to the circadian cycle, all administrations were performed simultaneously. To determine the siRNA effect, intraocular pressures (IOPs) were measured as previously mentioned.

[0152] Since glaucomatous pathology suggests an increase in intraocular pressure, the aim was to obtain a reduction in intraocular pressure levels following siRNA administration.

[0153] Most of the results for the different targets showed a significant reduction in IOP levels compared to controls and also with commercial drugs (latanoprost and dorzolamide) and animals treated with vehicle alone (negative controls) had a significant decrease in their IOP baseline There are no significant chan...

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Abstract

Sequences and protocols for treatment of eye conditions by use of RNA interference are disclosed. Target genes are selected from those responsible for aqueous flow or aqueous outflow, while particularly preferred conditions to be treated include glaucoma and uveitis.

Description

field of invention [0001] The present invention relates to methods and compositions useful in the treatment of eye diseases; particularly, though not exclusively, to the treatment of glaucoma. In a preferred embodiment, the present invention relates to the use of RNAi technology to down-regulate aqueous formation genes and aqueous outflow genes. Methods and compositions for treating eye diseases are also provided. Background of the invention [0002] RNAi as a tool to downregulate gene expression [0003] Gene targeting by homologous recombination is commonly used to determine gene function in mammals, but is a cost and time consuming method. Alternatively, the function of many genes can be determined following mRNA inhibition with ribozyme or antisense technology. Although successful in some cases, these techniques are difficult to apply universally. The advent of siRNA-directed "knockdown" has revolutionized somatic genetics, allowing cheap and rapid analysis of gene f...

Claims

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Application Information

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IPC IPC(8): C12N15/11A61P27/06
CPCC12N2310/14C12N15/1137C12N15/1138
Inventor 安娜・I・希门尼斯安吉拉・塞斯托何塞・P・罗曼伊雷妮・加斯孔贡萨洛・冈萨雷斯・德・布伊特拉格
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