KR-102962433-B1 - Composition for preventing, improving, or treating the mitochondrial complex 1 associated diseases comprising recombinant adeno-associated virus vector loading Hydra NDi1

Abstract

The present invention relates to a composition for the prevention, improvement, or treatment of mitochondrial complex 1 abnormalities, comprising a recombinant adeno-associated virus vector loaded with Hydra NDi1. The inventors have confirmed that the scAAV-hydra NDi1 of the present invention has the effect of restoring the reduction of cellular respiration caused by rotenone and the effect of inhibiting the death of LHON patient-derived cells caused by rotenone. It is expected that this can be used for the prevention, improvement, or treatment of diseases caused by abnormalities in mitochondrial complex 1 as well as LHON.

Inventors

• 기창원
• 손성수
• 박경아

Assignees

• 사회복지법인 삼성생명공익재단

Dates

Publication Date

20260508

Application Date

20230630

Claims (7)

1. A pharmaceutical composition for the prevention or treatment of Leber's hereditary optic neuropathy (LHON), comprising a self-complementary adeno-associated virus (scAAV) vector loaded with the hydra NDi1 gene represented by SEQ ID NO. 1.
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4. In paragraph 1, The above vector is a pharmaceutical composition for the prevention or treatment of Leber hereditary optic neuropathy that inhibits the reduction of cellular respiration.
5. In paragraph 1, The above vector is a pharmaceutical composition for the prevention or treatment of Leber hereditary optic neuropathy that inhibits apoptosis.
6. In paragraph 1, A pharmaceutical composition for the prevention or treatment of Leber hereditary optic neuropathy, wherein the above-mentioned Leber hereditary optic neuropathy has a mutation in one or more genes selected from the group consisting of ND1 (NADH dehydrogenase subunit 1), ND4 (NADH dehydrogenase subunit 4), and ND6 (NADH dehydrogenase subunit 6).
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Description

Composition for preventing, improving, or treating the mitochondrial complex 1 associated diseases comprising recombinant adeno-associated virus vector loading Hydra NDi1 The present invention relates to a composition for the prevention, improvement, or treatment of mitochondrial complex 1 abnormalities, comprising a recombinant adeno-associated virus vector loaded with Hydra NDi1. Mitochondrial genetic diseases caused by mutations in the mitochondrial genome were first reported about 30 years ago. Since then, there has been a dramatic advancement in our understanding of the numerous diseases resulting from these mutations and in mitochondrial genetics, which is involved in the transmission of mitochondrial genomes. Approximately thousands of mitochondrial genomes exist within a cell, and most mutated genomes exist intermingled with normal genomes; this is referred to as heteroplasmy. The severity of disease symptoms varies depending on the proportion of mutated genomes; symptoms become more severe as the proportion of mutated genomes—that is, as the heteroplasmy level increases. Identical mitochondrial genome mutations result in highly diverse heteroplasmies depending on the specific tissue or individual, and much research is still required to elucidate the mechanisms that regulate heteroplasmy. Leber's hereditary optic neuropathy (LHON) is a mitochondrial genetic disorder characterized by sudden, painless bilateral vision loss due to distinctive optic nerve damage occurring in young adults. It was first described in 1871 by the German ophthalmologist Theodore Leber. It manifests more frequently in men than women (approximately 80%), although women tend to experience later onset and are more severely affected. Onset typically occurs between the ages of 18 and 30, but it can appear without warning at any age, leading to simultaneous or sequential vision loss in both eyes, either acutely or subacutely. The exact incidence rate is not yet known. LHON was the first genetic disorder discovered to be inherited through mutations in mitochondrial genes (mitochondrial DNA, mtDNA), and it is inherited exclusively through the maternal line. While most patients have a family history from their mother, 40% of cases occur spontaneously. Mitochondria are important intracellular bodies that produce the energy (ATP) used for biological activities, and unlike nuclear genes, they possess independent genes. While most genetic diseases arise from defects in genes within the nucleus, some can manifest due to abnormalities in mitochondrial genes. Although a pair of genes within the cell nucleus is inherited from each parent, mitochondrial genes are inherited only from the maternal side because, during fertilization, the mitochondria containing paternal genes detach from the sperm's tail and cannot enter the egg. LHON accounts for 30–50% of optic neuropathy of unknown etiology occurring in both eyes and 2% of legal blindness. Approximately 90% of LHON patients possess one of three point mutations: G11778A (about 70% globally), G3460A, or T14484C (most common in people of mixed French and Canadian ancestry), and several other point mutation genes have also been identified. Most patients possess a single mutated gene (homoplasic), but cases with two or more mutated genes (heteroplasic) are rare. Although the exact mechanism of the disease's development has not been elucidated, it is believed that defective mitochondrial genes involved in normal cell division and growth processes disrupt energy production in optic nerve cells, leading to cell destruction. Although the administration of vitamin C, vitamin B12 , or idebenone as drug treatments can alleviate damage to retinal ganglion cells in patients with the disease, it has limitations in that it is not widely used in clinical practice due to its minimal effect and cannot provide fundamental prevention. Therefore, there is currently no suitable treatment available, and only gene therapy can be a fundamental cure. Accordingly, the inventors produced a recombinant viral vector containing hydra NDi1 to effectively treat mitochondrial disorders, and completed the present invention by experimentally demonstrating that mitochondrial disorders can be prevented, improved, and treated using this. Figure 1 is a schematic diagram showing the production of ssAAV-2/DJ-GFP, ssAAV-2/DJ-IRES-GFP, or scAAV-2/DJ-GFP recombinant adeno-associated virus. Figure 2 is a schematic diagram showing the production of ssAAV-2/DJ-yeast NDi1, ssAAV-2/DJ-hydra NDi1, and scAAV-2/DJ-hydra NDi1 recombinant adeno-associated viruses. Figure 3a shows the cleavage map of the recombinant adeno-associated virus vector used in experiments to confirm the gene expression intensity and stability of ssAAV and scAAV. Figure 3b shows the results of confirming the green fluorescence expression levels of ssAAV-GFP and scAAV-GFP. Figure 4 shows the results of adding rotenone to skin fibroblasts derived from LHON patients infected w