| G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease -> Monitor Keywords |
|
|
  G-rich polynucleotides as a novel therapeutic for the treatment of huntington's diseaseUSPTO Application #: 20080051363Title: G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease Abstract: The present invention relates to oligonucleotide compositions and therapeutic uses thereof to modify protein-protein interactions. In particular, the invention relates to the use of a guanidine-rich oligonucleotides to disrupt disease-causing protein aggregates, for example, Huntington's Disease (HD) protein aggregates (end of abstract) Agent: Mccarter & English, LLP Basil S. Krikelis - Wilmington, DE, US Inventors: Eric B. Kmiec, Hetal Parekh-Olmedo USPTO Applicaton #: 20080051363 - Class: 514044000 (USPTO) Related Patent Categories: Drug, Bio-affecting And Body Treating Compositions, Designated Organic Active Ingredient Containing (doai), O-glycoside, , Nitrogen Containing Hetero Ring, Polynucleotide (e.g., Rna, Dna, Etc.) The Patent Description & Claims data below is from USPTO Patent Application 20080051363. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part of U.S. patent application Ser. No. 11/544,278, filed Oct. 6, 2006, which claims the benefit under 35 U.S.C. .sctn.119(e) of U.S. Provisional Patent Application No. 60/724,085, filed Oct. 6, 2005, entitled: G-Rich Polynucleotides as a Novel Therapeutic for the Treatment of Huntington's Disease; the disclosures of which are hereby incorporated by reference in their entirety. INCORPORATION BY REFERENCE OF SEQUENCE LISTING [0002] The present application hereby incorporates by reference, in its entirety, the Sequence Listing, and identical CRF of the Sequence Listing filed herewith. The CRF contains nucleic acid sequences, SEQ. ID NO. 1-7, in file: "GRO_Kmiec.txt;" created: Oct. 4, 2006; OS: MS Windows XP; Software: PatentIn 3.3; size: 6 KB. The information contained in the Sequence Listing submitted, herewith, in the instant application is identical to the sequence information contained in the computer readable form. FIELD OF THE INVENTION [0003] The present invention relates to oligonucleotide compositions and methods of use thereof to modify protein-protein interactions. In particular, the invention relates to the use of a guanosine-rich oligonucleotides to disrupt disease-causing protein aggregates, for example, Huntington's Disease (HD) protein aggregates. BACKGROUND [0004] Huntington's Disease (HD) is an inherited autosomal dominant genetic disorder caused by expansions of CAG repeats (polyglutamine-polyQ) at the N-terminus, within exon 1, of the HD protein. The expansion of CAG repeats results in an increase in the length of the poly(Q) tract in the huntingtin (Htt) protein, which leads to protein misfolding and results in changes its solubility and induces protein aggregation that is the hallmark of HD. [0005] Pathologically, HD is marked by neuronal tissue degeneration due to the development of the protein aggregates. Aggregation occurs in two general phases, nucleation and elongation, and therefore, agents designed to block either phase are being considered as potential therapeutics. Recent studies suggest that mutant Htt can nucleate protein aggregation and interfere with a multitude of normal cellular functions. [0006] The extent of polyglutamine expansion is correlated with the severity of the symptoms and their onset while the pathology of the disease and neuronal cell death are thought to be associated with protein misfolding and protein aggregation. These aggregates are usually seen in the nucleus but can also be found in the cytoplasm. Protein aggregates develop via a complex biochemical process with intermediates being visible during the process. PolyQ tracts within the pathogenic range induce a protein insolubility whereas Htt with nonpathogenic length maintains a measured degree of solubility. [0007] Consistent with the aggregate toxicity hypothesis, inhibition of aggregate formation has been shown to have beneficial effects on the progression of HD in the R6/2 mouse model. The implication of the polyQ aggregates in cytotoxicity validates them as targets for novel therapeutics. Despite the lack of details surrounding the molecular structure of the polyQ aggregates, high throughput screening for compounds that inhibit their formation have produced some promising results. Several compounds, including Congo Red and Clioquinol, have been reported to inhibit the aggregation process in the R6/2 mouse model but their neurotoxicity tempers enthusiasm. Thus, identifying molecules that show efficacy with minimal toxicity should be an important consideration in the search for HD therapeutics. [0008] Much of the focus on developing therapeutics that block aggregate formation comes from a wealth of data associating HD pathogenesis with the presence of cellular inclusion bodies. But, recent evidence from in vitro and in vivo studies suggest that Htt inclusions may not be toxic to the cell or lead to neuronal degeneration. In fact, Hayden and colleagues have created an exciting mouse model that shows no long term effect of Htt inclusions on behavior or viability. It may be true that inclusion bodies are neuroprotective and eliminating them may actually increase the potential for neurotoxicity. [0009] Intracellular aggregates of Htt have long been considered phenotypic evidence of the neurodegenerative disorder Huntington's Disease. It is, however, not clear how the appearance of such inclusion bodies relates to the pathogenesis of the disease. A number of model systems have been designed to screen for therapeutic agents that can inhibit aggregation. Some of these assays measure the inhibition of fusion protein aggregation, proteins containing a fragment of Htt (here, GST-Q58-Htn) and a marker/reporter protein, often eGFP. The Htt component of this fusion protein harbors an expanded polyQ stretch. [0010] To date, most therapeutic treatments for HD are merely palliative. As such, efforts to find a therapy for HD have focused on agents that disrupt or block the mutant Htt aggregation pathway. However, there exists an ongoing need in the art for compounds that are capable of therapeutic intervention at alternative points in the HD cascade that demonstrate clinically relevant efficacy, are safe, and that are relatively inexpensive to manufacture. SUMMARY [0011] The present invention is based in part on the surprising and unexpected discovery that certain oligodeoxynucleotides (ODNs) are capable of inhibiting or ameliorating the cellular events that lead to HD disease pathology, and therefore, represent a new and useful class of therapeutics for the treatment or prevention of HD. An additional advantage is that synthetic oligodeoxnucleotides (ODNs) provide a model category of reagents that can be produced in highly purified quantities in a cost-effective way, and because of their small size are typically well-tolerated. [0012] Recently, we showed that GROs can inhibit and/or reduce protein aggregation, for example, the aggregation of Htt that occurs with HD. See J. Mol. Neurosci. 2004;24(2):257-67. While not being limited to any particular theory, the inventors theorize that the GROs may directly interact with the Htt proteins to inhibit nucleation, elongation, or both. [0013] There is some support for the notion of a direct interaction between certain GROs and proteins. Previously, GROs have been shown to bind directly to STAT3 and disrupt its function by interacting with regions of the protein that enable dimerization. In another instance, GROs have been shown to interact with HIV integrase and block integration of the HIV into the host chromosome. In addition, it has been reported that treatment of tumor cells with GROs inhibits cell cycle progression by interfering with DNA replication, as opposed to normal skin cells that exhibited minimal disruption of the cell cycle when treated with the same GROs (Xu et al.). However, no one has previously reported the ability of certain GROs to disrupt or reduce protein aggregation in general or Htt protein aggregation specifically. [0014] Therefore, in certain aspects the invention relates to guanosine-rich ODNs (GROs) compositions and therapeutic formulations thereof that are useful for inhibiting and/or reducing Htt protein aggregation. In additional aspects, the invention relates to methods of treating or preventing a disease related to protein aggregation, for example HD, comprising administering a therapeutically effective amount of a composition comprising a GRO in combination with at least one of a pharmaceutically acceptable carrier, excipient or both. [0015] Additional advantageous features and functionalities associated with the compositions, methods, and processes of the present invention will be apparent from the drawings presented herein, as well as the detailed description which follows. The publications and other materials used herein to illuminate the background of the invention, and in particular cases, to provide additional details respecting the practice, are incorporated by reference, and for convenience are listed in the appended bibliography. BRIEF DESCRIPTION OF THE DRAWINGS [0016] FIG. 1 is a diagram of the biochemical model screening assay obtained from Wang et al. (2005) and illustrating the steps involved in the biochemical/immunoblotting assay. The fusion protein GST-Q58-Htn (20 .mu.g/ml) was mixed with thrombin (0.5 unit/.mu.g protein) for 30 minutes and the mixture centrifuged to remove aggregated protein. The soluble protein was mixed with an ODN in a 96-well PCR plate and incubated for 24 hours at room temperature (RT). SDS was added to a final concentration of 2% and the mixture heated at 99.degree. C. for 5 minutes. Filtration through a 0.22 micron acetate cellulose membrane filter was followed by detection of aggregated Q58-Htn fragment by immunoblotting with an antibody (HP1) and ECL. Quantitation was carried out using an ImageQuant program. The blot displays both positive and negative results--positions lacking a black spot indicate that aggregation was inhibited by the ODN. [0017] FIG. 2(A) is the DNA sequence of two G-rich ODNs that form the G-quartet structure (SEQ ID NOs: 1 and 2). (B) is a dot blot analysis of T40216 (SEQ ID NO:2) and T30923 (SEQ ID NO:1) activity on aggregation. The zero (0) hour control represents reactions that were stopped immediately after addition of the protein; 24-hour reactions carried out in the absence of the ODN and stopped after 24-hours of incubation; Congo red, level of aggregation 24 hours after addition of Congo Red (10 .mu.M). [0018] FIG. 3 is a bar graph depicting aggregation inhibition by GROs. Here, T40216 (SEQ ID NO:2) and T30923 (SEQ ID NO:1) and Congo Red are used: Data are presented from five independent reactions, as shown in (B) for each point with standard deviation. *, denotes significance p<0.05 as compared to Congo Red (control) as determined by a one way ANOVA with Tukey's post hoc test. Continue reading... Full patent description for G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease or other areas of interest. ### Previous Patent Application: Composition for attenuating neuropathic pain comprising a recombinant ventor expressing gad65 Next Patent Application: Novel human lysosomal protein and methods of its use Industry Class: Drug, bio-affecting and body treating compositions ### FreshPatents.com Support Thank you for viewing the G-rich polynucleotides as a novel therapeutic for the treatment of huntington's disease patent info. IP-related news and info Results in 2.45493 seconds Other interesting Feshpatents.com categories: Canon USA , Celera Genomics , Cephalon, Inc. , Cingular Wireless , Clorox , Colgate-Palmolive , Corning , Cymer , |
||
