WO2025170079A1 - 慢性炎症を抑制するための医薬組成物 - Google Patents

慢性炎症を抑制するための医薬組成物

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Publication number
WO2025170079A1
WO2025170079A1 PCT/JP2025/004430 JP2025004430W WO2025170079A1 WO 2025170079 A1 WO2025170079 A1 WO 2025170079A1 JP 2025004430 W JP2025004430 W JP 2025004430W WO 2025170079 A1 WO2025170079 A1 WO 2025170079A1
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WO
WIPO (PCT)
Prior art keywords
mir
cells
hsa
pharmaceutical composition
derived
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/JP2025/004430
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English (en)
French (fr)
Japanese (ja)
Inventor
友宏 寺尾
隆 黒田
美砂 楠本
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cell Factor Inc
Original Assignee
Cell Factor Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cell Factor Inc filed Critical Cell Factor Inc
Priority to JP2025540402A priority Critical patent/JPWO2025170079A1/ja
Publication of WO2025170079A1 publication Critical patent/WO2025170079A1/ja
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
    • A61K35/545—Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives

Definitions

  • the present invention relates to a pharmaceutical composition for suppressing chronic inflammation.
  • Patent Document 1 discloses a therapeutic preparation for osteoarthritis or ligament or tendon injury, which contains a filtrate of adipose tissue-derived stem cell lysate as an active ingredient.
  • Patent Document 2 discloses a skin protective agent containing, as an active ingredient, a filtrate of a disruption solution obtained by subjecting adipose tissue-derived SVF fraction stem cells to disruption treatment, wherein the filtrate contains one or more proteins selected from the group consisting of heat shock proteins, cytokines and/or paracrine factors, and cell surface proteins, and the filtrate is a filtrate obtained by filtering only through a filter having a pore size of 0.1 ⁇ m to 0.45 ⁇ m.
  • the present invention aims to provide new pharmaceutical compositions and other substances that are effective in suppressing chronic inflammation.
  • RNA derived from iPS cells or adipose tissue-derived stem cells is effective in suppressing chronic inflammation, leading to the completion of the present invention.
  • a pharmaceutical composition for suppressing chronic inflammation comprising small RNA derived from iPS cells or adipose tissue-derived stem cells.
  • the present invention may be embodied as follows.
  • [13] A protoplasmic composition derived from iPS cells or adipose tissue-derived stem cells, from which the nucleus has been removed and which contains small RNA.
  • [14] The protoplast composition according to [13], wherein the iPS cells or adipose tissue-derived stem cells are human cells.
  • [17] The protoplast composition according to any one of [13] to [16], which is a cell lysate.
  • [E2] Use of the small RNA according to any one of [1] to [12], the pharmaceutical composition according to any one of [1] to [12], or the protoplast composition according to any one of [13] to [18] in the manufacture of a chronic inflammation inhibitor.
  • a chronic inflammation suppressing agent comprising small RNA derived from iPS cells or adipose tissue-derived stem cells, or a protoplasmic composition derived from iPS cells or adipose tissue-derived stem cells from which the nucleus has been removed and which contains small RNA.
  • a chronic inflammation inhibitor comprising the small RNA according to any one of [1] to [12], the pharmaceutical composition according to any one of [1] to [12], or the protoplast composition according to any one of [13] to [18].
  • the iPS cells are not particularly limited.
  • the upper limit of the number of subcultures is not particularly limited, but may be 20 times or less, 19 times or less, 18 times or less, 17 times or less, 16 times or less, 15 times or less, 14 times or less, 13 times or less, or 12 times or less.
  • the adipose tissue-derived stem cells of the present invention are preferably adipose tissue-derived stem cells that have reached a confluence of 75% or more, and more preferably 75% to 95%, or 80% to 90%.
  • the adipose tissue-derived stem cells of the present invention may be detached from adipose tissue-derived stem cells that have reached 75% or more, 75% to 95%, or 80% to 90% confluence and further subcultured to obtain adipose tissue-derived stem cells that have reached 75% or more, 75% to 95%, or 80% to 90% confluence again.
  • the multiple times of confluence may be two, three, four, five, or six times.
  • the pharmaceutical composition is used to suppress chronic inflammation.
  • suppression of chronic inflammation may mean treating or alleviating diseases or symptoms known as chronic inflammation, may mean exhibiting an inflammation suppressing effect, or may mean preventing diseases or symptoms known as chronic inflammation.
  • the pharmaceutical composition may be a chronic inflammation suppressant, may be a pharmaceutical composition for use in the treatment or prevention of chronic inflammation, or may be a pharmaceutical composition for use in the treatment or prevention of a disease or symptom associated with chronic inflammation.
  • the pharmaceutical composition may be a pharmaceutical composition for use in alleviating a disease or symptom associated with chronic inflammation.
  • RNA was extracted and purified from the homogenized sample using PureLink miRNA Isolation Kit (Invitrogen). Total RNA was dissolved in Nuclease-Free Water (Invitrogen). Each procedure was performed according to the manual.
  • EAE autoimmune encephalomyelitis
  • Cell lysis solution RIPA Buffer (Wako) 3x Sample Buffer 187.5mM Tris-HCl (pH 6.8), 6% SDS, 30% Glycerol Electrophoresis gel e-PAGEL (gradient gel, 10-20%) Buffer for Iemmli: SDS Tris-glycine buffer.
  • CBB staining solution 0.1% CBB, 30% methanol, 10% acetic acid.
  • Destaining solution 30% methanol, 10% acetic acid.
  • Detection antibody primary antibody
  • the cell lysate obtained in this example is considered to contain the small RNA obtained in Example 1, it is considered that the pharmacological effect in an experimental autoimmune encephalomyelitis (EAE) animal model can be confirmed by performing an evaluation experiment in an EAE animal model in the same manner as in Example 1.
  • EAE experimental autoimmune encephalomyelitis
  • Adipose tissue-derived stem cell lysates were prepared as follows: The adipose tissue-derived cells used were serial number PT-5006 (manufactured by Lonza). 1-1 Cell Culture 1-1-1 Frozen Cells Human Adipose-Derived Stem Cells x 1 Type (1 x 10 ⁇ 6 or more/vial, Transport Temperature: -80°C) (Manufactured by Lonza) 1-2 Extraction and purification of small RNA 1-2-1 Extraction reagent ISOSPIN Liquid Sample miRNA (Nippon Gene Co., Ltd.) Nuclease-Free Water (Invitrogen) DynaMarker RNA Low II (manufactured by BioDynamics Laboratory) NanoDrop One (Thermo Fisher Scientific) 2 Test Materials 2-1 Cell Culture 2-1-1 Cell Culture Medium (for human adipose-derived stem cells) ADSC-Adipose-Derived Stem Cells Growth Medium BulletKitTM (manufactured
  • Test Method 3-1 Cell Culture (Human Adipose-Derived Stem Cells) 3-1-1. Reactivation of frozen cells 1) Frozen cell stock was thawed in a 37°C water bath. 2) Suspended in culture medium. 3) Centrifuge at 200g for 3 minutes at room temperature. 4) The supernatant was removed, and the cells were suspended in culture medium and seeded onto a 100 mm or 150 mm culture dish at a cell density of approximately 5 x 10 3 cells/cm 2 . 5) The cells were cultured at 37°C in the presence of 5% CO2 . 3-1-2 Subculture 1) It was confirmed that the cells had proliferated to approximately 80-90% confluence.
  • the cells were seeded in a 150 mm culture dish containing 25 mL of culture medium at a cell density of approximately 5 x 103 cells/ cm2 . 9) The cells were cultured at 37°C in the presence of 5% CO2 . 10) Subculture was repeated until the number of cells reached 9.23 x 107 cells or more (because cell growth was slow after the second passage, the medium was changed to Mesenchymal Stem Cell Growth Medium 2 at the third passage. After the medium change, the growth rate increased from approximately 1.3-fold/day to approximately 2.3-fold/day). 3-1-3 Recovery and freezing of cells 1) It was confirmed that the cells in 3-1-2 had proliferated to approximately 80-90% confluence.
  • a cell suspension containing 6.03 x 10 7 cells was collected in a 50 mL tube. 9) Centrifuge at 200g for 3 minutes at room temperature. 10) The supernatant was removed, and the cell pellet was suspended in cell washing solution to a concentration of 4 ⁇ 10 6 cells/mL, and stored frozen at ⁇ 80° C. until use in step 3-2. 11) The remaining cell suspension was centrifuged at 200 g for 3 minutes at room temperature. 12) The supernatant was removed by aspirating, and a cryopreservation solution was added to adjust the cell density to 1.07 x 10 7 cells/mL. 13) The cell suspension was dispensed into cryovials in 1 mL portions. 14) Stored frozen at -80°C.
  • Example 4 The following experiment was also carried out in accordance with the evaluation of the effect of the present invention on the experimental autoimmune encephalomyelitis (EAE) model in Example 1.
  • the subjects (groups) to be administered are shown in Table 1 below.
  • the drug administration solution was administered on Days 2, 6, 10, and 14 as emulsion administration (Day 0).
  • the drug administration solution was administered on Days 2 and 6, and the vehicle administration solution was administered on Days 10 and 14.
  • a group in which the drug administration solution was administered 5 days and 2 days before elicitation was designated the two-times administration before elicitation group.
  • the average score on days 20-21 was lower than the average score at the peak.
  • the effect of the present invention was confirmed in an experimental autoimmune encephalomyelitis (EAE) model.
  • Example 5 In the same manner as in Example 1, to evaluate the effect of the present invention on the experimental autoimmune encephalomyelitis (EAE) model, the following experiment was also carried out by administering the compound on Day 14. When adipose tissue-derived cells were administered at 1.0 x 10 cells/animal, a further improvement in score was observed when cell lysate of adipose tissue-derived cells was administered in combination at 0.932 ⁇ g/animal (derived from 6.03 x 10 cells: 8 animals x 2 groups (16 animals)). The results are shown in Figure 3. Although the data is not disclosed, even in the group administered with cell lysate of adipose tissue-derived cells, the average score decreased from the peak average score on days 20 and 21.
  • EAE experimental autoimmune encephalomyelitis
  • hsa-miR-29c-3p-479229_mir hsa-miR-135a-5p-478581_mir hsa-miR-187-3p-477941_mir hsa-miR-196b-5p-478585_mir hsa-miR-197-3p-477959_mir hsa-miR-198-478749_mir hsa-miR-338-3p-478037_mir hsa-miR-504-5p-478144_mir hsa-miR-548a-5p-479501_mir hsa-miR-570-3p-479053_mir hsa-miR-589-5p-479073_mir hsa-miR-597-5p-478339_mir
  • the following seven miRNAs were confirmed to be expressed in common in iPS cells and adipose tissue-derived cells (undetermined in hDPSC cells).
  • hsa-miR-135a-5p-478581_mir hsa-miR-187-3p-477941_mir hsa-miR-197-3p-477959_mir hsa-miR-338-3p-478037_mir hsa-miR-504-5p-478144_mir hsa-miR-570-3p-479053_mir hsa-miR-589-5p-479073_mir
  • the following 18 miRNAs were not expressed in both iPS cells and adipose tissue-derived cells (although their expression was confirmed in hDPSC cells).

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Pulmonology (AREA)
  • Epidemiology (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Dermatology (AREA)
  • Rheumatology (AREA)
  • Reproductive Health (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Gynecology & Obstetrics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Pain & Pain Management (AREA)
  • Neurosurgery (AREA)
  • Otolaryngology (AREA)
  • Neurology (AREA)
  • Hematology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Urology & Nephrology (AREA)
  • Biochemistry (AREA)
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  • Heart & Thoracic Surgery (AREA)
PCT/JP2025/004430 2024-02-09 2025-02-10 慢性炎症を抑制するための医薬組成物 Pending WO2025170079A1 (ja)

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JP2019516771A (ja) * 2016-05-25 2019-06-20 エラスムス ユニバーシティ メディカルセンター ロッテルダムＥｒａｓｍｕｓ Ｕｎｉｖｅｒｓｉｔｙ Ｍｅｄｉｃａｌ Ｃｅｎｔｅｒ Ｒｏｔｔｅｒｄａｍ 間葉系幹細胞およびその一部の使用
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JP2021528984A (ja) * 2018-07-02 2021-10-28 リン、シー−ランLIN, Shi−Lung 成体幹細胞の拡大と誘導のインビトロでの誘発
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JP2023533556A (ja) * 2020-07-10 2023-08-03 シャリテ－ウニヴェルジテーツメディツィン・ベルリン 心臓修復のためのマイクロｒｎａ標的療法
JP2022045325A (ja) * 2020-09-08 2022-03-18 デクソンファーマシューティカルズ株式会社 サイトカインストーム抑制剤、サイトカインストーム抑制剤の使用方法およびスクリーニング方法
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