CN-121971315-A - Use of recombinant extracellular vesicle bioactive protein delivery systems for improving cellular oxidative stress

Abstract

The present disclosure provides the use of catalase or a variant or nucleic acid sequence or vector or delivery system thereof for the preparation of a pharmaceutical or cosmetic composition for scavenging Reactive Oxygen Species (ROS), alleviating oxidative stress.

Inventors

• LI MING
• CHEN YIYOU

Assignees

• 艾思曼生物科技有限公司

Dates

Publication Date

20260505

Application Date

20260202

Priority Date

20250311

Claims (18)

1. 1. Use of catalase or a variant or nucleic acid sequence or vector or delivery system thereof for the preparation of a pharmaceutical or cosmetic composition for scavenging Reactive Oxygen Species (ROS), alleviating oxidative stress.
2. 2. The use according to claim 1, wherein the catalase or variant thereof has at least the biological activity of scavenging Reactive Oxygen Species (ROS), alleviating oxidative stress.
3. 3. The use according to claim 1 or 2, wherein the catalase or variant thereof is a wild-type catalase or a modified catalase, wherein the modified catalase is selected from at least one of the group consisting of: (i) Wild-type catalase having one or more amino acid mutations, insertions, deletions and/or additions, or (Ii) Wild-type catalase modified with chemical groups.
4. 4. A use according to any one of claims 1-3, wherein the catalase or variant thereof is selected from at least one of the group consisting of: (i) A catalase having an amino acid sequence as set forth in SEQ ID No. 1, or a catalase having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% sequence identity to SEQ ID No. 1.
5. 5. The use according to any one of claims 1 to 4, wherein the nucleic acid sequence encodes a catalase or variant thereof as defined in any one of claims 2 to 4.
6. 6. The use according to any one of claims 1 to 5, wherein the vector comprises a nucleic acid sequence as defined in claim 5.
7. 7. The use according to any one of claims 1-6, wherein the delivery system comprises: (i) A catalase or variant thereof as defined in any one of claims 2-4, and/or, (Ii) A nucleic acid sequence as defined in claim 5 or a vector as defined in claim 6; (iii) A delivery vehicle, wherein the delivery vehicle has the function of delivering catalase or a variant thereof into a specific tissue or cell.
8. 8. The use of claim 7, wherein the delivery vehicle is selected from at least one of a liposome, an Extracellular Vesicle (EV), and a virus-like particle (VLP).
9. 9. The use according to claim 7 or 8, wherein the delivery vehicle is selected from at least one of a generic extracellular vesicle or a modified extracellular vesicle, Alternatively, the extracellular vesicles are universal exosomes or modified exosomes.
10. 10. The use of claim 9, wherein the extracellular vesicles are harvested from production cells; Alternatively, the extracellular vesicles are exosomes, and the exosomes may be modified to enhance production, exposure duration, tissue-specific targeting, or endosomal escape; More optionally, the exosomes comprise (i) peptides and/or proteins comprising GPI-anchored signal sequences, (ii) peptide/antibody fragment modifications, and/or (iii) other protein modifications.
11. 11. The use according to claim 10, wherein the producer cells overexpress at least one protein or protein fragment selected from the group consisting of CD46, CD52, CD55, CD58 and CD 59.
12. 12. The use according to claim 10 or 11, wherein the producer cell is a non-human mammalian cell line or a human cell line.
13. 13. The use according to any one of claims 10-12, wherein the producer cell is selected from a HEK 293F cell line, a HEK 293T cell line, a stem cell line or any combination thereof.
14. 14. Use according to any one of claims 1 to 13, wherein the delivery system is formed and secreted/released into the culture medium by production cells transfected with the vector as defined in claim 6 and produced intracellularly, followed by isolation and purification.
15. 15. The use of claim 14, wherein the producer cell is selected from a HEK 293F cell line, a HEK 293T cell line, a stem cell line, or any combination thereof.
16. 16. The use according to any one of claims 1 to 15, wherein the pharmaceutical composition comprises: (i) A catalase or a variant thereof as defined in any one of claims 2-4, or, (Ii) The nucleic acid sequence as defined in claim 5 or the vector as defined in claim 6, or, (Iii) A delivery system as defined in any one of claims 7 to 15, and (Iv) Pharmaceutically acceptable excipients.
17. 17. Use according to any one of claims 1 to 15, characterized in that the cosmetic composition comprises: (i) A catalase or a variant thereof as defined in any one of claims 2-4, or, (Ii) The nucleic acid sequence as defined in claim 5 or the vector as defined in claim 6, or, (Iii) A delivery system as defined in any one of claims 7 to 15, and (Iv) Acceptable excipients.
18. 18. The use according to any one of claims 1-17, wherein the pharmaceutical or cosmetic composition is for reducing ROS levels in skin, alleviating oxidative stress damage, and/or treating vitiligo.

Description

Use of recombinant extracellular vesicle bioactive protein delivery systems for improving cellular oxidative stress Cross reference The present application claims priority from PCT application No. PCT/CN2025/081856 filed on day 3 and 11 of 2025. The content of the prior PCT application is considered part of the present disclosure and is incorporated herein in its entirety. Technical Field The present disclosure relates to the field of biological medicine, in particular to the use of recombinant extracellular vesicle bioactive protein delivery systems in ameliorating cellular oxidative stress. Background Skin aging and related cellular aging are caused by exogenous factors (e.g., ultraviolet irradiation, smoking, pollution) and endogenous factors (e.g., time, genetic factors, hormones). Ultraviolet (UV) radiation is the strongest external driving factor for age-related changes in skin (i.e. "photoaging"). Facial aging is about 80% attributable to photoaging. Skin photoaging is clinically manifested mainly as photoelastic tissue degeneration, wrinkles and pigmentation. Ultraviolet radiation triggers the overproduction of Reactive Oxygen Species (ROS) in the epidermis and dermis of the skin, mitochondria being the primary site of ROS production in cells. ROS negatively affect skin cells, including keratinocytes and fibroblasts, by inducing oxidative damage and promoting inflammatory responses. These effects can lead to decreased cell viability, accelerated collagen degradation, and up-regulation of pro-inflammatory cytokines. Various features of skin aging are associated with inflammation. Elevated ROS levels are typically observed during polarization of M0 macrophages to pro-inflammatory M1 macrophages. At the same time, excessive ROS may further promote M1 polarization. Cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) are markers of M1 macrophages, whose levels are found to be upregulated in photoaged skin. The therapeutic use of small molecules is undesirable because they do not effectively scavenge intracellular reactive oxygen species. The reaction speed of antioxidant enzyme and these oxidants is thousands to millions times faster than that of small molecules, and is the main antioxidant defense mechanism. Catalase, as the most active antioxidant enzyme in nature and in the human body, plays a vital role in relieving oxidative stress by effectively scavenging ROS. The ability of catalase to decompose hydrogen peroxide into water and oxygen significantly reduces oxidative damage caused by ultraviolet radiation. This enzymatic activity not only protects keratinocytes and fibroblasts from ROS-induced cell damage, but also helps to preserve the integrity and elasticity of the skin. Catalase acts as an effective strategy for improving visible signs of photoaging, such as wrinkles and pigmentation. Extracellular Vesicles (EVs) are lipid bilayer nanoparticles that are released from all cell types into the extracellular space. EVs naturally transport the necessary cellular components (e.g., proteins or active enzymes, lipids, nucleic acids such as mRNA, micro-RNA) for intercellular communication. Thus, EVs hold great promise in delivering active ingredients, payloads, or therapeutic agents to target cells or within cells. As a natural secretion product of cells, EVs have numerous advantages as drug delivery systems/vectors compared to synthetic drug delivery vectors (e.g., liposomes, lipid nanoparticles, and viral vectors). EVs are naturally derived from cells, making them inherently biocompatible and less prone to eliciting immune responses than synthetic vectors. This property is critical to reducing potential side effects in therapeutic applications, EVs have inherent tissue targeting and the ability to cross biological barriers, EVs exhibit extremely high stability in biological fluids, which helps to extend circulation time in extracellular matrix and blood. EVs have been widely validated in scientific research and clinical applications, and their effectiveness and safety have been demonstrated in the field of medical cosmetology. Potential therapeutic applications include anti-aging, anti-pigmentation, wound healing, hair regrowth, and the like. However, the active ingredients in natural stem cell derived EVs are very limited. Catalase has a short half-life after entering the body due to rapid proteolytic degradation and immunogenicity. Thus, encapsulation of catalase in extracellular vesicles enhances its stability, extends its effective half-life, allows for better penetration into target tissues and cells, and provides for efficient scavenging of reactive oxygen species therein. Currently, a variety of methods have been developed for loading therapeutic agents into EVs, including sonication, electroporation, and passive incubation. However, these conventional methods involve multiple manufacturing steps, including catalase protein expression, EV preparation, and loading