阅读说明：本技术 用于调节τ蛋白表达的组合物 (Compositions for modulating tau protein expression ) 是由 H·柯达赛维茨 E·E·斯韦兹 S·M·弗赖尔 H·布维 于 2014-07-21 设计创作，主要内容包括：本文公开了用于降低τ蛋白mRNA和蛋白质表达的反义化合物和方法。这种方法、化合物和组合物可用于治疗、预防或改善τ蛋白相关疾病、病症和病状。(Disclosed herein are antisense compounds and methods for reducing tau mRNA and protein expression. Such methods, compounds, and compositions are useful for treating, preventing, or ameliorating tau protein-associated diseases, disorders, and conditions.)

1. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19 or at least 20 consecutive nucleobases of any one of the nucleobase sequences SEQ ID NOS 20-2443 and SEQ ID NO 2478 and 2483.

2. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the nucleobase sequences SEQ ID NOs 446, 313, 321, 1634, and 2309.

3. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of SEQ ID NO 446.

4. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of SEQ ID NO 313.

5. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of SEQ ID NO 321.

6. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of SEQ ID NO 1634.

7. The compound of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of SEQ ID NO 2309.

8. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19 or at least 20 consecutive nucleobases of any one of the nucleobase sequences SEQ ID NO 2444 and 2484 and 2565.

9. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any one of the nucleobase sequences SEQ ID NOs 20-2565.

10. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising an equal length portion of at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19 or at least 20 contiguous nucleobases complementary to nucleobases 135783 and 135980 of SEQ ID NO: 1.

Technical Field

Compositions and methods for reducing expression of tau mRNA and protein in an animal are provided. Such methods are useful for treating, preventing or ameliorating neurodegenerative diseases including tauopathies, Alzheimer's Disease, frontotemporal dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), chronic traumatic brain Disease (CTE), corticobasal ganglionic degeneration (CBD), epilepsy and Dravet's Syndrome by inhibiting the expression of tau protein in an animal.

The major function of tau is to bind and stabilize microtubules, which are important cytoskeletal structural components involved in mitosis, cytokinesis and vesicle transport. Tau protein is found in a variety of tissues, but is particularly abundant in axons of neurons. In humans, there are six tau isoforms produced by alternative splicing exons 2, 3 and 10. Splicing exons 2 and 3 at the N-terminus of the protein results in the inclusion of 0,1 or 2 acidic domains of 29 amino acids, and are referred to as 0N, 1N or 2N tau protein, respectively. The effect of these domains on tau function is not completely clear, but may play a role in interactions with the plasma membrane. Inclusion of exon 10 at the C-terminus results in inclusion of the microtubule binding domain encoded by exon 10. Since there are 3 microtubule-binding domains elsewhere in the tau protein, this isoform of tau (including exon 10) is called 4R tau protein, where 'R' refers to the number of repeats of the microtubule-binding domain. Tau without exon 10 is called 3R tau. Since more microtubule binding domains (4R compared to 3R) would increase binding to microtubules, 4R tau protein is postulated to significantly increase microtubule binding and assembly. The ratio of 3R/4R tau protein is developmentally regulated, with fetal tissue expressing only 3R tau protein and adult human tissue expressing approximately equal amounts of 3R/4R tau protein. Deviations from the normal ratio of 3R/4R tau are characteristic of neurodegenerative FTD tau pathology. It is unknown how varying the 3R/4R tau ratio at a later stage in adult animals will affect tau pathogenesis.

Serine-threonine directed phosphorylation regulates the microtubule binding ability of tau protein. Hyperphosphorylation promotes tau detachment from microtubules. Other post-translational modifications of tau have been described; however, the meaning of these modifications is ambiguous. Phosphorylation of tau is also developmentally regulated, with higher phosphorylation in fetal tissues and much lower phosphorylation in adults. One characteristic of neurodegenerative disorders is abnormally increased tau phosphorylation.

The microtubule network is involved in many important processes (including structural integrity) within the cell that are required to maintain the morphology of the cell and to manipulate transport mechanisms. Tau protein may be a key mediator of some of these processes and disruption of normal tau protein in neurodegenerative diseases may disrupt some of these key cellular processes, since tau protein binding to microtubules stabilizes microtubules.

One early indication that tau protein may be important in neurodegenerative syndromes is the recognition that tau protein is a key component of the content of nerve fibers in alzheimer's disease. In fact, the nerve fiber content is an aggregate of hyperphosphorylated tau. Together with plaques containing beta amyloid, the nerve fiber content is a hallmark of alzheimer's disease and is significantly associated with cognitive impairment. The accumulation of 95% tau protein in AD is found in neuronal processes and is called neuroinflammatory dystrophy. The process by which this microtubule-associated protein becomes derived from microtubule detachment and formation of protein accumulation and how this is associated with neuronal toxicity is not well understood.

Neuronal tau inclusion is not only a pathological feature of alzheimer's disease, but also a subset of frontotemporal dementia (FTD), PSP and CBD. The association between tau and neurodegeneration was consolidated by finding that mutations in the tau gene would cause a subset of FTDs. These genetic data have also emphasized the importance of the 3R:4R ratio of tau protein. Many tau mutations that cause FTD result in tau splicing changes that result in preferential inclusion of exon 10, and thus an increase in 4R tau. The overall tau protein content was normal. It remains unknown whether changes in tau isoforms or amino acids or both lead to neurodegeneration. Recent data indicate that PSP can also be correlated with an increase in the 4R:3R tau protein ratio.

To help understand the effect of tau ratio on neurodegeneration, pocket-sized genes comprising the tau promoter and flanking intron sequences of exon 10 have been used to generate mouse models based on a splice tau mutation (N279K). As in humans, these mice show increased levels of 4R tau protein compared to transgenes expressing WT tau protein, and develop behavioral and motor abnormalities and accumulation of aggregated tau protein in the brain and spinal cord.

The protein "tau" has been associated with a variety of brain diseases including Alzheimer's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal ganglion degeneration, dementia pugilistica, chromosome-associated Parkinson's disease (parkinsonism), Lytico-Bodig's disease (Lytico-Bodig's disease), tangle-predominant dementia (tangle-predominant dementia), ganglioglioma, gangliocytoma, meningioangiomatosis, subacute sclerosing panencephalitis, plumbagic encephalopathy, tuberous sclerosis, Harlervorden-Spatz disease (Hallervorden-Spatz disease), Pick's disease, argentious granule disease (argyrophilic gain disease), corticobasal degeneration or frontotemporal degeneration, and other diseases. Tau-associated disorders such as AD are the most common cause of dementia in the elderly. AD affects an estimated 1500 million people worldwide and 40% of the population is over the age of 85. AD is characterized by two pathological hallmarks: tau nerve fiber inclusions (NFT) and beta amyloid (Α β) plaques.

There is currently a lack of acceptable options for treating such neurodegenerative diseases. It is therefore an object herein to provide methods for treating such diseases.

Summary of The Invention

Provided herein are methods, compounds, and compositions for modulating tau protein mRNA and protein expression. In certain embodiments, the compounds useful for modulating tau mRNA and protein expression are antisense compounds. In certain embodiments, the antisense compound is an antisense oligonucleotide.

In certain embodiments, modulation may be in a cell or tissue. In certain embodiments, the cell or tissue is in an animal. In certain embodiments, the animal is a human. In certain embodiments, tau mRNA levels are reduced. In certain embodiments, tau protein content is reduced. This reduction may occur in a time-dependent manner or in a dose-dependent manner.

Also provided are methods, compounds, and compositions useful for preventing, treating, and ameliorating diseases, disorders, and conditions. In certain embodiments, such tau-associated diseases, disorders, and conditions are neurodegenerative diseases. In certain embodiments, such neurodegenerative diseases, disorders, and conditions include tauopathy, alzheimer's disease, frontotemporal dementia (FTD), FTDP-17, Progressive Supranuclear Palsy (PSP), chronic traumatic brain disease (CTE), corticobasal ganglionic degeneration (CBD), epilepsy, and Dravet syndrome.

Such diseases, disorders, and conditions may have one or more risk factors, etiologies, or outcomes in common. Some risk factors and etiologies for developing neurodegenerative disorders include aging, having a personal or family history, or genetic predisposition. Certain symptoms and outcomes associated with the development of neurodegenerative disorders include, but are not limited to: hyperphosphorylated tau protein, presence of nerve fiber inclusions, reduced neurological function, reduced memory, reduced motor function, reduced motor coordination and confusion.

In certain embodiments, the methods of treatment comprise administering to an individual in need thereof a tau protein antisense compound. In certain embodiments, the method of treatment comprises administering to an individual in need thereof a tau protein antisense oligonucleotide.

The present disclosure provides the following non-limiting numbered embodiments:

embodiment 1: a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19 or at least 20 consecutive nucleobases of any one of the nucleobase sequences SEQ ID NOS 20-2443 and SEQ ID NO 2478-2483.