KR-102960618-B1 - Pharmaceutical composition for preventing or treating bone disease using peptide comprising N-acetylcysteine

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of bone diseases using a peptide containing N-acetylcysteine. More specifically, it was confirmed that a peptide containing N-acetylcysteine, a cell-permeable peptide, and a nuclear membrane-permeable peptide increased bone matrix and collagen fiber bundles, reduced ROS and DNA damage, and increased the expression of CD31 and RUNX2 in a mouse model of periodontitis, thereby allowing for various applications such as therapeutic agents for bone diseases and compositions for health functional foods.

Inventors

• 김재영
• 하정홍
• 이상일
• 장제희
• 박정현
• 김안나
• 김태영
• 손원주
• 성시진
• 안서영
• 안창현
• 정재광

Assignees

• 경북대학교 산학협력단

Dates

Publication Date

20260507

Application Date

20240508

Claims (10)

1. A pharmaceutical composition for the prevention or treatment of periodontitis comprising, as an active ingredient, a peptide consisting of an amino acid sequence represented by SEQ ID NO. 1 or SEQ ID NO. 2.
2. delete
3. delete
4. In claim 1, The above composition is a pharmaceutical composition characterized by increasing ALP activity.
5. In claim 1, The above composition is a pharmaceutical composition characterized by increasing the expression of bone matrix or collagen fiber bundles.
6. In claim 1, The above composition is a pharmaceutical composition characterized by increasing the expression of the CD31 or RUNX2 gene.
7. delete
8. A health functional food composition for preventing or improving periodontitis comprising, as an active ingredient, a peptide consisting of an amino acid sequence represented by SEQ ID NO. 1 or SEQ ID NO. 2.
9. delete
10. A reagent composition for increasing CD31 or RUNX2 expression comprising, as an active ingredient, a peptide consisting of an amino acid sequence represented by SEQ ID NO. 1 or SEQ ID NO. 2.

Description

Pharmaceutical composition for preventing or treating bone disease using a peptide comprising N-acetylcysteine The present invention provides a pharmaceutical composition for the prevention or treatment of bone diseases using a peptide containing N-acetylcysteine. Teeth are composed of hard tissues—enamel, dentin, and cementum—and soft tissue, the pulp. Enamel is a hard tissue that protects the tooth by resisting wear applied to the crown. Because enamel is located on the outermost layer of the tooth, it is susceptible to damage from external stimuli such as dental caries, trauma, and wear. Nevertheless, since the ameloblasts that form tooth enamel disappear immediately after tooth eruption, remodeling or self-healing by ameloblasts is difficult once the enamel is damaged. Dentin constitutes the majority of the tooth and plays a pivotal functional role; it is organized to resist masticatory pressures applied to the tooth in a complementary manner to enamel. Although dentin is less hard than enamel, it possesses flexibility that compensates for enamel's brittle nature. Dentin forms the pulp-dentin complex together with the pulp, which is composed of loose connective tissue. The odontoblasts that form dentin differentiate from undifferentiated mesenchymal cells of neural crest origin and are highly polar, well-organized cells arranged along the dentin-pulp interspace. Unlike bone, which undergoes continuous remodeling throughout a lifetime, dentin is considered a tissue that is very difficult to regenerate due to its unique anatomical and physiological characteristics, in which it is not remodeled once lost and there is no replacement tissue. To regenerate damaged dentin, undifferentiated mesenchymal cells present in the pulp must be differentiated into odontoblasts, similar to the developmental process. Representative factors known to date for odontoblast differentiation and dentin formation include TGF-β, Wnt, bone morphogenetic protein (BMP), Sonic Hedgehog (Shh), and fibroblast growth factor (Fgf). As such, while teeth do not spontaneously regenerate in adults, they are tissues that are continuously exposed to the outside and used in daily life, making them organs that can be easily damaged or lost due to cavities, periodontal disease, or external impact. Damage to or loss of teeth not only affects an individual's chewing, pronunciation, aesthetics, and psychological well-being but also has significant social and economic repercussions. In the past, teeth were often replaced using methods such as dentures or implants, but now there is a need for research on developing new technologies to biologically regenerate damaged tooth structures, moving beyond conventional treatment methods. Figure 1 shows the structure of N-acetylcysteine and the peptide sequences of NAC-CPP and NAC-NLS. Figure 2 shows the evaluation of cytotoxicity in MC3T3 cell lines through MTS analysis. Figure 3 shows the evaluation of ALP activity in MC3T3 cell lines after NAC-NLS treatment. Figure 4 shows the results of HE and MTC staining after one week of NAC-NLS treatment in a mouse model of periodontitis. Figure 5 shows the presence of ROS and DNA damage after one week of NAC-NLS treatment in a mouse model of periodontitis. Figure 6 shows the expression patterns of CD31, an angiogenesis marker, and RUNX2, a precursor osteoblast marker, after staining one week after NAC-NLS treatment in a mouse model of periodontitis. Figure 7 shows the results of Micro-CT imaging taken 2 weeks after NAC-NLS treatment in a mouse model of periodontitis. The present invention will be described in more detail below. The present invention provides a pharmaceutical composition for the prevention or treatment of bone disease comprising a peptide consisting of an amino acid sequence represented by SEQ ID NO. 1 as an active ingredient. The C-terminus of the above peptide may be amideated or the N-terminus may be acetylated. The above composition may be a peptide comprising an amino acid sequence represented by SEQ ID NO. 2, which further includes an amino acid sequence represented by SEQ ID NO. 3 in addition to a peptide comprising an amino acid sequence represented by SEQ ID NO. 1. The above composition can increase ALP activity. The above composition can increase the expression of bone matrix or collagen fiber bundles. The above composition can increase the expression of the CD31 or RUNX2 gene. The above bone diseases may be one or more selected from the group consisting of gingivitis, periodontitis, tooth loss, dental caries, peri-implantitis, dentin-pulp disease and traumatic fracture of the tooth, osteoarthritis, rheumatoid arthritis, osteoporosis and osteomalacia, but are not limited thereto. In another embodiment of the present invention, the pharmaceutical composition may further comprise one or more additives selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, leavening agents, lubricants, lubri