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KR-102961550-B1 - Pharmaceutical composition containing cell-derived natural or artificial nanovesicles loaded with anti-sense oligonucleotide-based drugs

KR102961550B1KR 102961550 B1KR102961550 B1KR 102961550B1KR-102961550-B1

Abstract

The present invention relates to a pharmaceutical composition capable of introducing a nucleic acid-based drug into a cell by loading the nucleic acid-based drug onto a cell-derived natural or artificial nanovesicle, and a method for manufacturing the same. According to the present invention, natural or artificial nanovesicles loaded with antisense oligonucleotides exhibit the characteristics of inhibiting the proliferation and migration of cancer cells and increasing drug sensitivity when administered in combination with existing chemotherapy drugs.

Inventors

  • 이재용
  • 안희진
  • 이대한
  • 이동열
  • 나규흠

Assignees

  • 주식회사 강스템테라퓨틱스

Dates

Publication Date
20260507
Application Date
20220630
Priority Date
20210630

Claims (16)

  1. An antisense oligonucleotide (ASO) designed to target a sequence within the Galectin-3 coding sequence (CDS) and modified in which at least one base in the sequence, either partially or entirely, is modified with a phosphorothioate and/or 2'-O-methyl group to enhance stability within the cell.
  2. A cell-derived natural or artificial nanovesicle loaded with an antisense oligonucleotide (ASO) designed using a base sequence within a Galectin-3 coding sequence (CDS) as a target, wherein the antisense oligonucleotide (ASO) is characterized in that some or all of at least one base in the base sequence is modified with a phosphorothioate and/or 2'-O-methyl group.
  3. delete
  4. A cell-derived natural or artificial nanovesicle loaded with an antisense oligonucleotide (ASO) designed using a base sequence within a Galectin-3 coding sequence (CDS) as a target, wherein the antisense oligonucleotide (ASO) is characterized in that some or all of at least one base in the base sequence is modified with a phosphorothioate and/or 2'-O-methyl group.
  5. In paragraph 2, the natural or artificial nanovesicles are cell-derived natural or artificial nanovesicles characterized by being secreted or prepared from human embryonic kidney cells or umbilical cord blood stem cells.
  6. delete
  7. A cell-derived natural or artificial nanovesicle characterized by being for the prevention or treatment of cancer in any one of paragraphs 2, 4, and 5.
  8. delete
  9. A cell-derived natural or artificial nanovesicle characterized in that, in any one of claims 2, 4, and 5, the antisense oligonucleotide (ASO) is designed using one or more sequences of SEQ ID NOs 1 to 3 for Galectin-3 targeting.
  10. Cell-derived natural or artificial nanovesicles characterized by being formulated for oral or nasal inhalation or intravenous injection in accordance with claim 2, 4, or 5.
  11. A cell-derived natural or artificial nanovesicle characterized by being formulated for the prevention or treatment of lung cancer in any one of paragraphs 2, 4, and 5.
  12. A cell-derived natural or artificial nanovesicle characterized by being administered in combination with a non-nucleic acid-based drug in any one of paragraphs 2, 4, and 5.
  13. A cell-derived natural or artificial nanovesicle characterized by reducing the proliferative and/or motility of target cells in any one of paragraphs 2, 4, and 5.
  14. A cell-derived natural or artificial nanovesicle characterized by increasing sensitivity to an anticancer agent administered in combination, in any one of paragraphs 2, 4, and 5.
  15. delete
  16. A Galectin-3 targeting antisense oligonucleotide (ASO) according to claim 1, characterized by being designed using one or more sequences of SEQ ID NOs 1 to 3.

Description

Pharmaceutical composition containing cell-derived natural or artificial nanovesicles loaded with antisense oligonucleotide-based drugs The present invention relates to a pharmaceutical composition characterized by containing cell-derived natural or artificial nanovesicles loaded with a nucleic acid-based drug. For example, the present invention relates to the development of an anticancer drug utilizing human cell-derived natural or artificial nanovesicles loaded with antisense oligonucleotides against cancer-inducing gene base sequences. Cancer is classified according to the location in the body organ where it originates, such as liver cancer, stomach cancer, or uterine cancer. Cancer that develops in the lungs is called lung cancer, or more precisely, primary lung cancer. In other words, it refers to cancer that arises in lung tissues such as the bronchi, bronchioles, and alveoli. In contrast, cases where cancer originates in other organs—such as breast or liver cancer—and metastasizes to the lungs are called metastatic lung cancer and are treated differently from primary lung cancer. Lung cancer has the highest probability of causing death from cancer. Since it progresses slowly, it can be treated surgically if detected early. Because the clinical course and treatment differ, lung cancer is classified into Non-Small Cell Lung Cancer (NSCLS) and Small Cell Lung Cancer (SCLC) based on the microscopic size and morphology of the cancer cells; NSCLS accounts for 85% of lung cancer patients. NSCLS is further divided into Non-Squamous Non-Small Cell Lung Cancer (NSCLC) and Squamous Cell Lung Cancer (SCLC). Non-Squamous NSCLC accounts for the majority, at 70–75%, while Squamous NSCLC accounts for approximately 25%. Although some drugs currently used as targeted anticancer agents for NSCLC demonstrate high therapeutic efficacy, there is currently no treatment method for resistant cancers caused by the EGFR T790M mutation. All cells exchange information with other cells or the external environment. To facilitate this information exchange, cells secrete various substances into the external environment, including growth factors, cytokines, chemokines, hormones, and neurotransmitters. Recently, in addition to these single substances, information exchange via extracellular vesicles (EVs) is receiving increasing attention. Bacterial exosomes may contain lipopolysaccharides or lipoteichoic acid. Extracellular vesicles are nanoscale vesicles secreted by all cells into the external environment for intercellular information exchange. Extracellular vesicles contain various substances exhibiting biological activity, such as proteins, lipids, nucleic acids, and metabolites. Furthermore, the components of extracellular vesicles reflect the type and state of the cell from which they originate and exist in various bodily fluids, including cell culture media, blood, urine, saliva, tears, semen, breast milk, ascites, and cerebrospinal fluid. Extracellular vesicles derived from various cells can perform diverse physiological and pathological functions by interacting with target cells. For example, stem cell-derived extracellular vesicles can induce tissue regeneration, while cancer cell-derived vesicles can influence cancer growth, metastasis, and angiogenesis. Bacterial-derived extracellular vesicles can perform functions such as antibiotic resistance, elimination of competing bacteria, delivery of virulence factors, and regulation of immune responses. Meanwhile, there are proteins commonly found in extracellular vesicles derived from various cells and body fluids, which confirms that proteins do not enter the extracellular vesicles in a disorderly manner but rather through regulated mechanisms. Analysis of extracellular vesicle proteomes based on their organelles of origin revealed that proteins derived primarily from the plasma membrane or cytosol were abundant, while proteins derived from mitochondria or the cell nucleus were relatively scarce. Proteins commonly found in extracellular vesicles are involved in their structure, formation, and transport; representative examples include tetraspanins (CD9, CD63, CD81, etc.), integrins, endosomal sorting complex proteins (TSG101, Alix), cytoskeletal proteins (actins, tubulin), and heat shock proteins (HSP70, HSP90). Proteins commonly found in extracellular vesicles derived from mammalian cells can be utilized as marker proteins when isolating extracellular vesicles. Meanwhile, extracellular vesicles also contain cell-type specific proteins, which are associated with performing physiological or pathological functions related to the cell type. Extracellular vesicles are composed of various biologically active substances and can perform diverse biological functions. Mechanisms by which extracellular vesicles interact with target cells include ligand-receptor interactions, fusion, and endocytosis. Extracellular vesicles derived from various cells interact with target cells, performing ph