KR-20260065643-A - Composition for preventing or treating muscular atrophy or muscle damage comprising CU-rich RNA

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of muscular dystrophy or muscle damage or a composition for promoting muscle differentiation, comprising as an active ingredient a CU-rich RNA derived from the ChRO1 gene, which is a long non-coding RNA, or a homomorphism thereof. The composition has the effect of maintaining the staining central structure of muscle cells, reducing the expression of proteolytic factors, and increasing the expression of muscle activity-related factors, and thus can be used more safely and effectively for the prevention or treatment of muscular dystrophy or muscle damage.

Inventors

• 조은정
• 최진미
• 박상하

Assignees

• 성균관대학교산학협력단

Dates

Publication Date

20260511

Application Date

20241031

Claims (14)

1. A pharmaceutical composition for the prevention or treatment of muscular dystrophy or muscle damage comprising CU-rich RNA as an active ingredient.
2. In Article 1, A pharmaceutical composition in which the above CU-rich RNA is derived from the ChRO1 gene, which is a long noncoding RNA, or an isoform thereof.
3. In Article 1, A pharmaceutical composition wherein the above CU-rich RNA comprises a CU dinucleotide repeat sequence and complementary sticker sequences at both ends of the repeat sequence.
4. In Paragraph 3, A pharmaceutical composition in which the above CU dinucleotide repeat sequence and complementary sticker sequence are directly connected or connected by a linker sequence.
5. In Paragraph 3, A pharmaceutical composition in which the above CU dinucleotide repeat sequence is a sequence of length 100 to 900 nucleotides (nt).
6. In Paragraph 3, A pharmaceutical composition wherein the above CU dinucleotide repeat sequence comprises the nucleotide sequence of SEQ ID NO. 5 or SEQ ID NO. 10.
7. In Paragraph 3, A pharmaceutical composition wherein the above complementary sticker sequence is a sequence of length 9 to 12 nucleotides (nt).
8. In Paragraph 3, A pharmaceutical composition wherein the above complementary sticker sequence comprises the nucleotide sequences of SEQ ID NO. 6 and SEQ ID NO. 7, or the nucleotide sequences of SEQ ID NO. 11 and SEQ ID NO. 12.
9. In Article 1, A pharmaceutical composition in which the above CU-rich RNA forms a loop structure.
10. In Article 1, A pharmaceutical composition wherein the above CU-rich RNA comprises a nucleotide sequence selected from the group consisting of SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 8, and SEQ ID NO. 9.
11. In Article 1, A pharmaceutical composition in which the above-mentioned muscular atrophy is caused by decreased muscle function, muscle wasting, or muscle degeneration.
12. In Article 1, A pharmaceutical composition wherein the above-mentioned muscle injury is selected from the group consisting of muscle strain, muscle rupture, muscle tearing, contusion, distortion, rotator cuff syndrome, and myositis.
13. A composition for promoting muscle differentiation containing CU-rich RNA as an active ingredient.
14. A method for the prevention or treatment of muscular dystrophy or muscle damage comprising the step of administering a pharmaceutically effective amount of CU-rich RNA to an individual.

Description

Composition for preventing or treating muscular atrophy or muscle damage comprising CU-rich RNA The present invention relates to a pharmaceutical composition for the prevention or treatment of muscular atrophy or muscle damage or a composition for promoting muscle differentiation, comprising as an active ingredient CU-rich RNA derived from the ChRO1 gene, which is a long non-coding RNA, or a homomorphism thereof. Muscular dystrophy, characterized by a decline in skeletal muscle mass and function, is a myopathy caused by various factors. Muscles atrophy and muscle strength decline due to diseases of the muscles themselves, immobility resulting from limb fixation and bed rest, and malnutrition, as well as severe diseases such as aging, degenerative neurological disorders, acquired immunodeficiency syndrome (AIDS), chronic heart failure, sepsis, and cancer. Patients with muscular dystrophy experience a significant decline in quality of life due to limited movement caused by muscle loss, potentially reaching a point where they cannot even utilize the minimum muscles necessary for long-term survival. Furthermore, when muscular dystrophy accompanies severe diseases, the prognosis is poor, as it affects treatment methods and effectiveness and increases mortality rates. Therefore, while active treatment for muscular dystrophy alongside the disease is necessary, there are currently no effective preventive or therapeutic methods other than exercise and nutritional therapy. In particular, since exercise and nutritional therapy are difficult to apply to the elderly with muscular dystrophy and patients with severe diseases, the development of preventive and therapeutic substances is even more critical. RNA therapeutics are treatments that use RNA (ribonucleic acid) molecules to treat or prevent diseases. Unlike DNA-based therapies, they are considered safer as there is no risk of insertion into the host cell genome. Furthermore, RNA therapeutics possess high selective specificity as they can act on target molecules that cannot be administered as small molecules. Unlike DNA-based therapies, which must penetrate the cytoplasm and nuclear membrane, RNA therapeutics have the advantage of being able to regulate the activity of intracellular proteins simply by entering the cytoplasm. In addition, they can be developed more rapidly compared to protein therapeutics, such as small molecules or monoclonal antibodies, resulting in relatively lower development costs. Accordingly, the inventors completed the present invention by confirming that CU-rich RNA derived from the ChRO1 gene, which is a long noncoding RNA, or its isoform is effective in preventing or treating muscular dystrophy. Figure 1a shows the results of sequencing analysis of muscle-specifically expressed ChRO1 RNA (left: mouse ChRO1, mChRO1; and right: human ChRO1, hChRO1), 1b shows the results of ChIRP-seq data analysis of DNA sequences to which ChRO1 is bound in C2C12 myotube cells (GSE94498 for ChRO1; and GSE113248 for MyoDeRNA as a control), 1c shows the results of analyzing the sequences of mChRO1a and hChRO1a expressed in parts of exon1 and intron1 of ChRO1, 1d shows the results of structural prediction of mChRO1a and hChRO1a, and 1e shows the results of predicting the protein binding site of mChRO1a. Figure 2 shows the results of performing in vitro phase separation experiments using HP1a protein after dividing and cloning several fragments of ChRO1a and transcribing them with IVT. Figure 3a shows the results of observing the heterochromatin foci pattern with and without 1,6-HD treatment in C2C12 myoblasts capable of overexpressing CU-rich 413nt RNA (CUR 413nt), and Figure 3b shows the results of confirming the intracellular locations of chromatin proteins through immunostaining in C2C12 myoblasts capable of overexpressing CU-rich 413nt RNA (CUR 413nt). Figure 4a is an experimental schedule in which C2C12 myoblasts capable of overexpressing CU-rich 413nt RNA (CUR 413nt) were differentiated into myotubes and then treated with Dox and Dex for 2 days for analysis; 4b is the result of analyzing changes in heterochromatin aggregation patterns and the number of foci through DAPI staining; 4c is the result of confirming the expression levels of the proteolytic factors ATROGIN1 and MuRF1 ; 4d is the result of analyzing the thickness of myotubes; 4e is the result of performing RNA-seq analysis on muscle samples from adult males (RNA-seq data: PRJNA450495); and 4f is the result of analyzing the correlation between the expression of ChRO1 and genes. Figure 5a is the result of analyzing the sequence of human ChRO1a, 5b is the result of predicting the structure of human ChRO1a, 5c is a schematic diagram of an RNA molecule consisting of a CU-rich sequence and a sticker sequence capable of complementary binding to the ends, 5d is a CU-rich RNA molecule derived from the human ChRO1a sequence modified according to the RNA molecule schematic diagram, and 5e is the result of pre