WO2019168364A1 - Procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches - Google Patents

Procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches Download PDF

Info

Publication number
WO2019168364A1
WO2019168364A1 PCT/KR2019/002408 KR2019002408W WO2019168364A1 WO 2019168364 A1 WO2019168364 A1 WO 2019168364A1 KR 2019002408 W KR2019002408 W KR 2019002408W WO 2019168364 A1 WO2019168364 A1 WO 2019168364A1
Authority
WO
WIPO (PCT)
Prior art keywords
stem cells
thioredoxin
cells
pharmaceutical composition
ischemic brain
Prior art date
Application number
PCT/KR2019/002408
Other languages
English (en)
Korean (ko)
Inventor
박원순
장윤실
성동경
Original Assignee
사회복지법인 삼성생명공익재단
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
Priority claimed from KR1020190023053A external-priority patent/KR102242040B1/ko
Application filed by 사회복지법인 삼성생명공익재단 filed Critical 사회복지법인 삼성생명공익재단
Priority to US16/977,524 priority Critical patent/US20210000877A1/en
Publication of WO2019168364A1 publication Critical patent/WO2019168364A1/fr

Links

Images

Classifications

    • 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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor

Definitions

  • the present invention relates to a method for enhancing the expression of thioredoxin (TRX) of stem cells, and more particularly a method for enhancing the expression of thioredoxin in stem cells comprising the step of culturing stem cells in a hypoxic environment
  • TRX thioredoxin
  • the present invention relates to stem cells having enhanced thioredoxin expression by the above method, and to the use of the stem cells for ischemic brain disease treatment.
  • Thioredoxin is a low molecular weight protein with a molecular weight of 10,000 to 13,000 and was found as a coenzyme that donates hydrogen ions to ribonucleotide reductase, an enzyme essential for DNA synthesis in Escherichia coli.
  • Thioredoxin has an active site called -Cys-Gly-Pro-Cys- and is an oxidized form to form disulfide (SS) bond between two cysteine residues and a reduced form to form dithiol (-SH-SH). It is a redox control factor in the cells present.
  • Thioredoxin has been reported to have a number of biological activities, such as cells that act as growth factors and eliminate intracellularly toxic hydrogen peroxide and affect NF- ⁇ B signaling activity in eukaryotic cells, thereby affecting these signaling. It is reported to affect death and tumors. Because of this activity, thioredoxin has recently attracted attention in the field of anti-cancer drug development and antioxidant protein protection against cell damage.
  • ischemic brain disease is a type of cerebrovascular disease that can be subdivided into thrombosis, embolism, transient ischemic attack, and infarction, mainly due to thrombus and embolism. It refers to a disease caused by pathological abnormalities in blood vessels that supply blood flow to the brain.
  • transient cerebral ischemia in the cerebrum, the supply of oxygen and glucose is blocked, resulting in a decrease in ATP and edema in neurons, resulting in extensive damage to the brain.
  • Neuronal death occurs after a significant period of time after cerebral ischemia, which is called delayed neuronal death.
  • Neuronal cell death caused by cerebral ischemia occurs in two main mechanisms. One is that excess glutamate accumulates outside the cell by cerebral ischemia and this glutamate flows into the cell, causing neuronal cell death due to excessive intracellular calcium accumulation. Excitatory neuronal death mechanism, and another is the oxidative neuronal death mechanism caused by damage to DNA and cytoplasm due to the increase of radicals in vivo due to the sudden supply of oxygen during ischemia-reperfusion.
  • Drugs currently available for the treatment of ischemic brain disease are antithrombotic, antiplatelet or anticoagulants such as ticlopidine, cilostazole and prostacycline are often burdened with headaches, palpitations and liver. It has been reported to have limitations in use due to side effects.
  • the FDA-issued commercial tissue plasminogen activator dissolves the blood clots that induce cerebral ischemia and induces rapid oxygen and glucose supply, but it does not directly protect neurons and therefore requires early use. Due to its thrombolytic effect, excessive or frequent use causes thinning of the blood vessel walls, resulting in hemorrhagic cerebrovascular disease (Int J Stroke. 2014 Apr; 9 (3): 349-55).
  • the present invention has been made to solve the above-mentioned conventional problems, the present inventors observed that the expression of thioredoxin significantly increased when the stem cells were cultured in ischemic hypoxic conditions, unlike normal conditions specifically In addition, it was confirmed that the effect of inhibiting the death of neurons, thereby completing the present invention.
  • an object of the present invention is to provide a method of enhancing thioredoxin (TRX) expression of stem cells and a stem cell having enhanced expression of thioredoxin by the above method.
  • Another object of the present invention is to provide a pharmaceutical composition for treating ischemic brain disease, including stem cells having enhanced thioredoxin expression.
  • Another object of the present invention is to provide a thioredoxin delivery complex, including thioredoxin (TRX) and polydimethylsiloxane (PDMS) nanoparticles.
  • TRX thioredoxin
  • PDMS polydimethylsiloxane
  • Another object of the present invention is to provide a pharmaceutical composition for treating ischemic brain disease, comprising the complex.
  • the present invention provides a method for enhancing the expression of thioredoxin (TRX) of stem cells, comprising culturing the stem cells in a hypoxic environment.
  • TRX thioredoxin
  • the present invention provides a stem cell, the thioredoxin expression is enhanced by the above method.
  • the low oxygen environment may be an environment in which oxygen of 0.5 to 10% is supplied.
  • the stem cells in the group consisting of mesenchymal stem cells, human tissue-derived mesenchymal stromal cells, human tissue-derived mesenchymal stem cells, multipotent stem cells and amnion epithelial cells may be selected.
  • the mesenchymal stem cells may be derived from umbilical cord, umbilical cord blood, bone marrow, fat, muscle, nerve, skin, amniotic membrane, placenta, amniotic fluid or tonsil.
  • the present invention also provides a pharmaceutical composition for treating ischemic brain disease, including stem cells having enhanced thioredoxin expression.
  • the ischemic brain disease is neonatal hypoxic ischemic brain injury, stroke, cerebral infarction, cerebral ischemia, thrombosis, embolism, transient ischemic attack, lacune , Cerebral hemorrhage, head trauma, cerebral circulatory metabolic disorders, brain coma, traumatic brain injury, and hypoxic brain injury.
  • the present invention also provides a method for treating ischemic brain disease, comprising administering to a subject a pharmaceutical composition for treating ischemic brain disease comprising stem cells having enhanced thioredoxin expression.
  • the present invention provides a use of the pharmaceutical composition for the treatment of ischemic brain disease.
  • the present invention also provides a thioredoxin delivery complex, including thioredoxin (TRX) and polydimethylsiloxane (PDMS) nanoparticles.
  • TRX thioredoxin
  • PDMS polydimethylsiloxane
  • the nanoparticles may be prepared by mixing a tetraethyl orthosilicate (TEOS) and dimethyldiethoxysilane (DMDES), followed by a condensation reaction under a base catalyst.
  • TEOS tetraethyl orthosilicate
  • DMDES dimethyldiethoxysilane
  • the present invention also provides a pharmaceutical composition for treating ischemic brain disease, comprising the complex.
  • the present invention also provides a method for treating ischemic brain disease, comprising administering to a subject a pharmaceutical composition comprising the complex.
  • the present invention also provides a therapeutic composition for ischemic brain disease of the pharmaceutical composition comprising the complex.
  • the present inventors confirmed that when stem cells were cultured in hypoxic conditions, the expression of thioredoxin was significantly increased only in hypoxic conditions, unlike in normal conditions, and thiore in neurons induced injury under ischemic hypoxic environment. Apoptosis was inhibited, and the therapeutic effect of intra-ventricular administration of thioredoxin via delivery vehicle was confirmed in neonatal hypoxic ischemic encephalopathy in vivo model.
  • the stem cells may be usefully used for the prevention or treatment of ischemic brain disease.
  • Figure 1a is a result of confirming 61 kinds of proteins (OGD abundance) and their position in the cell is increased expression compared to the control stem cells (NC) from the culture of stem cells (OGD) cultured in hypoxic conditions.
  • Figure 1b is a result showing that the expression of thioredoxin (Thioredoxin) significantly increased in stem cells (OGD) cultured in hypoxic conditions compared to the control (NC).
  • Figure 2a is exposed to 1% oxygen concentration of the cerebral cortex-derived neurons in the rat fetus for 1 hour to induce cell damage and then treated with thioredoxin (OGD thioredoxin) compared with the untreated thioredoxin (OGD control) The results showed that apoptosis was significantly suppressed.
  • OGD thioredoxin thioredoxin
  • FIG. 2B shows cell survival after treatment with thioredoxin by concentration (1, 10, 100, 500, 1000 ng) or mesenchymal stem cells (MSC) in neurons cultured under the same conditions as in FIG. 2A. ) And free radical species (ROS) levels.
  • Figure 3a illustrates the process of transporter synthesis for the delivery of thioredoxin in neurons.
  • 3B is an in vitro result showing intracellular uptake of the carrier.
  • 3C shows ex vivo results showing the uptake of neurons in neurons and results showing uptake into brain cells upon intraventricular administration of rats.
  • Figure 4 is a result showing the improved therapeutic effect of damaged brain tissue during intraventricular administration of thioredoxin through the carrier in neonatal hypoxic ischemic encephalopathy in vivo model.
  • the present inventors observed that the expression of thioredoxin was significantly increased differently from normal conditions, and thus the neuronal cell death inhibition effect was confirmed.
  • the present invention has been completed.
  • the present invention provides a method for enhancing the expression of thioredoxin (TRX) of stem cells, comprising culturing the stem cells in a hypoxic environment.
  • TRX thioredoxin
  • the present invention provides a stem cell, the thioredoxin expression is enhanced by the above method.
  • the present inventors confirmed that the expression of thioredoxin is significantly enhanced as a result of culturing stem cells in a hypoxic environment. More specifically, when the culture medium of the stem cells cultured in hypoxic conditions (1 hour culture at 5% oxygen concentration and 6 hours at normal conditions) or normal conditions (7 hours culture at normal conditions) collected and cultured in hypoxic conditions As a result of examining the proteins with increased expression, 61 kinds of proteins were identified, and the expression level of thioredoxin protein was significantly increased (see Example 1).
  • the low oxygen environment may be an environment in which oxygen of 0.5 to 10% is supplied, more preferably may be an environment in which 1-5% of oxygen is supplied.
  • stem cell refers to a cell that is an undifferentiated cell and has the ability to differentiate into two or more different types of cells while having self-replicating ability.
  • Stem cells of the present invention may be autologous or allogeneic stem cells, may be from any type of animal, including humans and non-human mammals, and is not limited to whether the stem cells are derived from adults or embryos Do not.
  • the stem cells are selected from the group consisting of mesenchymal stem cells, human tissue-derived mesenchymal stromal cells, human tissue-derived mesenchymal stem cells, multipotent stem cells and amniotic epithelial cells More preferably, the stem cells of the present invention may be mesenchymal stem cells, the mesenchymal stem cells are derived from the umbilical cord, cord blood, bone marrow, fat, muscle, nerve, skin, amniotic membrane, placenta, amniotic fluid or tonsil It may be, but if derived from human in vivo tissue is not limited in its origin.
  • the present invention provides a pharmaceutical composition for treating ischemic brain disease, comprising stem cells with enhanced thioredoxin expression.
  • the effect of thioredoxin with increased expression from stem cells in a hypoxic environment was examined for neurons. More specifically, neurons isolated from the cerebral cortex of the rats were exposed to 1% oxygen for 1 hour to cause injury, then treated with thioredoxin and analyzed for cell viability. As a result, it was confirmed that the killing of neurons was significantly inhibited compared with the neurons not treated with thioredoxin, and this effect appeared in proportion to the treatment concentration of thioredoxin and the concentration of reactive oxygen species It was confirmed to decrease (see Example 2).
  • a carrier capable of delivering thioredoxin into cells was synthesized, and the carrier was confirmed to be absorbed into neurons. Furthermore, thioredoxin was included in a neonatal hypoxic ischemic encephalopathy in vivo model. As a result of administering the carrier into the ventricle, the treatment effect was confirmed to significantly improve the damaged brain tissue.
  • the ischemic brain disease may include neonatal hypoxic ischemic brain injury, stroke, cerebral infarction, cerebral ischemia, thrombosis, embolism, transient ischemic attack, lacune, cerebral hemorrhage, and head trauma. ), Brain circulation metabolic disorders, brain function coma, traumatic brain injury and hypoxic brain injury, but may be any one selected from the group consisting of, but is not limited thereto.
  • the present invention provides a thioredoxin delivery complex, including thioredoxin (TRX) and polydimethylsiloxane (PDMS) nanoparticles.
  • TRX thioredoxin
  • PDMS polydimethylsiloxane
  • the nanoparticles may be prepared by mixing a tetraethyl orthosilicate (TEOS) and dimethyldiethoxysilane (DMDES), and then condensation under a base catalyst, but the manufacturing method is not limited thereto. .
  • TEOS tetraethyl orthosilicate
  • DMDES dimethyldiethoxysilane
  • the present invention provides a pharmaceutical composition for treating ischemic brain disease, comprising the complex.
  • the pharmaceutical composition of the present invention may further contain one or more known auxiliary components having a therapeutic effect of ischemic brain disease together with stem cells or the complex having enhanced thioredoxin expression.
  • the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the treatment of ischemic brain diseases.
  • composition of the present invention may further comprise a suitable carrier commonly used in the manufacture of pharmaceutical compositions.
  • suitable carrier commonly used in the manufacture of pharmaceutical compositions.
  • Such pharmaceutically acceptable carriers are conventionally used in the preparation, and include, but are not limited to, saline solution, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, and the like. If necessary, other conventional additives such as antioxidants and buffers may be further included.
  • diluents, dispersants, surfactants, binders, lubricants and the like may be additionally added to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.
  • Suitable pharmaceutically acceptable carriers and formulations can be preferably formulated according to the individual components using methods disclosed in Remington's literature.
  • the pharmaceutical composition of the present invention is not particularly limited in formulation, but may be formulated as an injection, inhalant, or external skin preparation.
  • compositions of the present invention can be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is determined by the condition and weight of the patient, Depending on the extent, drug form, route of administration, and time, it may be appropriately selected by those skilled in the art.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat or diagnose a disease at a reasonable benefit / risk ratio applicable to medical treatment or diagnosis, and an effective dose level refers to a patient's disease type, severity, or drug. Can be determined according to the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, factors including the drug used concurrently and other factors well known in the medical field.
  • the pharmaceutical compositions according to the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.
  • the present invention also provides a method for treating ischemic brain disease, comprising administering to a subject a pharmaceutical composition comprising stem cells with enhanced thioredoxin expression.
  • the term "individual” means a subject in need of treatment of a disease, and more specifically, a human or non-human primate, mouse, rat, dog, cat, horse, cow, and the like. Mean mammal.
  • the present invention provides a therapeutic use of the ischemic brain disease of the pharmaceutical composition.
  • the present inventors conducted this experiment to develop a stem cell therapy for ischemic brain disease, an ischemic hypoxic environment.
  • stem cells were incubated for 1 hour at an oxygen concentration of 5% and then cultured for 6 hours under normal conditions, and then cultured cells were collected. After culturing for only 7 hours under the conditions, the culture solution of the cells was collected. Next, each collected culture solution was centrifuged at 3000 rpm for 30 minutes to remove debris from the culture solution, and then the medium was concentrated for protein analysis. Subsequently, after trypsin digestion, fractionation was performed, and mass analysis was performed to analyze proteins having increased expression in stem cells exposed to hypoxic conditions compared to control stem cells.
  • the inventors of the present invention found that i) an antioxidant protein being studied as a therapeutic target for ischemic hypoxic brain disease, ii) the price of a synthetic protein is significantly lower than that of a recombinant protein, and iii) Finally, after selecting thiorredoxin as a protein that satisfies all conditions for easy development of various types of carriers, further analysis was performed. The result of changing the expression level of the thioredoxin in a hypoxic environment is shown in Figure 1b.
  • Example 1 Based on the results of Example 1, the present inventors tried to verify the effect of thioredoxin on neurons in an ischemic hypoxic environment. To this end, we examined the inhibitory effect of thioredoxin on neuronal cell death in hypoxic-induced neuronal cell death in vitro.
  • the embryos were taken out of the white paper of 18.5 days of gestational age and the cerebral cortex was separated to culture neurons. Subsequently, the neuron was exposed to 1% oxygen for 60 minutes to induce cell injury, and then, by treating thioredoxin with neuronal cells induced cell death, the cell death inhibition effect was observed. (dojindo, korea).
  • neuronal cell death was significantly induced in the neuronal cells exposed to hypoxic conditions and not treated with thioredoxin, compared to the control group cultured under normal conditions (NC).
  • Neurode treated with thioredoxin OGD thioredoxin was confirmed that the cell death was inhibited to a significant level.
  • thioredoxin was treated with various concentrations (1, 10, 100, 500, 1000 ng) or mesenchymal stem cells (MSC) for the neurons cultured under the same conditions as described above. Cell survival was measured by.
  • the reactive oxygen species assay kit was used to measure changes in free radicals and used as an evaluation tool for treatment efficacy.
  • ROS reactive oxygen species
  • the present inventors synthesized a carrier capable of delivering thioredoxin into cells according to the following method. More specifically, as shown in FIG. 3A, tetraethyl orthosilicate (TEOS) and dimethyldiethoxysilane (DMDES) are mixed and hydrolyzed, followed by condensation under a base catalyst. Through) to prepare a siloxane-based PDMS nanoparticle (siloxane-based PDMS nanoparticle) carrier. Then, to verify whether the delivery medium prepared by the method uptake in the cell.
  • TEOS tetraethyl orthosilicate
  • DMDES dimethyldiethoxysilane
  • Example 3-1 In order to verify the cellular uptake of the carrier prepared according to the method of Example 3-1, the following experiment was carried out. More specifically, warming the 37 ° C water bath and the frozen cell tube was rapidly dissolved in the water bath for 2-3 minutes, and then completely centrifuged for 5 minutes at room temperature at a rate of 300 xg when the cells completely dissolved. After centrifugation, the supernatant was removed, the cell pellet was placed in a DMEM culture medium, and once again centrifuged under the same conditions. The supernatant was removed, and the cell pellet was resuspended in the culture medium.
  • Example 3-2 Based on the results of Example 3-2, the present inventors further verified whether the delivery vehicle is delivered to neurons and brain.
  • the embryos were taken out of the white paper after 18 days of pregnancy and separated from the cerebral cortex, and then separated into single cells by treatment with trypsin and pipetting. Cells were then counted and seeded to a density of 20,000 cells / cm 2 and incubated in an incubator. When the cells were about 50% grown, 1 ⁇ l of the tetramethyltamine-containing carrier was added to the culture solution of the cultured cells and incubated for 24 hours. Then, the fluorescent microscope was used to observe whether the carrier was delivered into the cells.
  • hypoxic ischemic encephalopathy HIE
  • HIE hypoxic ischemic encephalopathy
  • Staining was performed by dissolving TTC powder in 37 ° C. saline to make a 2% w / v TTC solution, and then immersing the brain tissue in the solution and incubating at 37 ° C. for 15-30 minutes.
  • the damaged brain tissue was significantly improved when the thioredoxin-containing carrier was administered (20% DMDES thioredoxin) compared to the control without the thioredoxin-containing carrier (20% DMDES thioredoxin). It was confirmed that one therapeutic effect appeared.
  • the method according to the present invention it is possible to significantly enhance the expression of thioredoxin in stem cells, and in addition to the inhibitory effect of neuronal cell death by thioredoxin, thiore through a carrier in neonatal hypoxic ischemic encephalopathy in vivo model It has been experimentally confirmed that the therapeutic effect during the intraventricular administration of single cells, stem cells with enhanced thioredoxin expression in a hypoxic environment by the method according to the present invention can be useful in the field of treatment of ischemic brain disease. will be.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Developmental Biology & Embryology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Epidemiology (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Urology & Nephrology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Vascular Medicine (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches et, plus particulièrement, un procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches, le procédé comprenant une étape de culture de cellules souches dans une condition hypoxique, les cellules souches ayant une expression régulée à la hausse de thiorédoxine par le même procédé, et une utilisation des cellules souches dans le traitement d'une maladie cérébrale ischémique. Lorsqu'elles sont cultivées dans une condition hypoxique, des cellules souches sont autorisées à augmenter l'expression de thiorédoxine et de cellules souches qui ont un niveau d'expression régulé à la hausse de thiorédoxine présentant un excellent effet thérapeutique sur une maladie cérébrale ischémique peuvent être obtenues par le procédé. Ainsi, les cellules souches peuvent être utilement appliquées dans la prévention ou le traitement d'une maladie cérébrale ischémique.
PCT/KR2019/002408 2018-03-02 2019-02-28 Procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches WO2019168364A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US16/977,524 US20210000877A1 (en) 2018-03-02 2019-02-28 Method For Upregulation Of Thioredoxin Expression In Stem Cells

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2018-0025278 2018-03-02
KR20180025278 2018-03-02
KR10-2019-0023053 2019-02-27
KR1020190023053A KR102242040B1 (ko) 2018-03-02 2019-02-27 줄기세포의 티오레독신 발현을 증진시키는 방법

Publications (1)

Publication Number Publication Date
WO2019168364A1 true WO2019168364A1 (fr) 2019-09-06

Family

ID=67806318

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2019/002408 WO2019168364A1 (fr) 2018-03-02 2019-02-28 Procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches

Country Status (1)

Country Link
WO (1) WO2019168364A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115120616A (zh) * 2022-06-10 2022-09-30 中国人民解放军海军军医大学 miR-199a-5p过表达工程干细胞外泌体在制备治疗芥子气致肺损伤药物中的应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998032863A2 (fr) * 1997-01-28 1998-07-30 Karo Bio Ab Thioredoxine mammalienne
US20160331875A1 (en) * 2013-11-27 2016-11-17 Trustees Of Boston University Stretch release drug delivery materials
WO2017075136A1 (fr) * 2015-10-27 2017-05-04 Purdue Research Foundation Produits thérapeutiques à base de polymère pour le brunissement inductif de graisse

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998032863A2 (fr) * 1997-01-28 1998-07-30 Karo Bio Ab Thioredoxine mammalienne
US20160331875A1 (en) * 2013-11-27 2016-11-17 Trustees Of Boston University Stretch release drug delivery materials
WO2017075136A1 (fr) * 2015-10-27 2017-05-04 Purdue Research Foundation Produits thérapeutiques à base de polymère pour le brunissement inductif de graisse

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PARK, K.-J. ET AL.: "Expression Pattern of the Thioredoxin System in Human Endothelial Progenitor Cells and Endothelial Cells Under Hypoxic Injury", KOREAN CIRCULATION JOURNAL, vol. 40, no. 12, 2010, pages 651 - 658, XP055635344 *
TAKAGI, Y. ET AL.: "Overexpression of thioredoxin in i transgenic mice attenuates focal ischemic brain damage", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES (PNAS), vol. 96, no. 7, 1999, pages 4131 - 4136, XP055635346 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115120616A (zh) * 2022-06-10 2022-09-30 中国人民解放军海军军医大学 miR-199a-5p过表达工程干细胞外泌体在制备治疗芥子气致肺损伤药物中的应用
CN115120616B (zh) * 2022-06-10 2024-04-30 中国人民解放军海军军医大学 miR-199a-5p过表达工程干细胞外泌体在制备治疗芥子气致肺损伤药物中的应用

Similar Documents

Publication Publication Date Title
Li et al. Mn (III) meso-tetrakis-(N-ethylpyridinium-2-yl) porphyrin mitigates total body irradiation-induced long-term bone marrow suppression
US20100178277A1 (en) Methods and compositions for stimulating cells
WO2013025042A2 (fr) Composition comprenant des microvésicules issues de cellules souches pour favoriser la neurogenèse
WO2019151840A1 (fr) Composition pharmaceutique comprenant des mitochondries isolées pour la prévention ou le traitement de la polyarthrite rhumatoïde
WO2014157918A1 (fr) Composition pharmaceutique pour inhiber la réponse immunitaire par induction de la différenciation en cellules t régulatrices et promouvoir la prolifération des cellules t régulatrices
WO2014036377A1 (fr) Procédés favorisant la viabilité cellulaire
WO2017146309A1 (fr) Nouvelle utilisation de l'eupatiline comme composition pharmaceutique pour prévenir et traiter la fibrose via la capacité de celle-ci à inhiber la transition épithélio-mésenchimateuse
Wang et al. A comparative study of Mesenchymal Stem Cells transplantation approach to antagonize age-associated ovarian hypofunction with consideration of safety and efficiency
JP2015159895A (ja) 脳梗塞治療のための多能性幹細胞
WO2021210872A1 (fr) Composition de prévention ou de traitement d'une maladie de la peau diabétique, comprenant un exosome dérivé de cellule souche traitée par de la thrombine
CA3052739A1 (fr) Nouvelles vesicules extracellulaires anti-angiogeniques
WO2020222483A1 (fr) Composition pharmaceutique pour le traitement de la septicémie ou du syndrome de réponse inflammatoire systémique, comprenant des mitochondries isolées en tant que principe actif
WO2013085303A1 (fr) Cellules souches multipotentes dérivées de la membrane amniotique canine
Li et al. Mild hypothermia facilitates mitochondrial transfer from astrocytes to injured neurons during oxygen-glucose deprivation/reoxygenation
WO2019168364A1 (fr) Procédé de régulation à la hausse de l'expression de thiorédoxine dans des cellules souches
Sefton et al. Fibroblast-derived Hgf controls recruitment and expansion of muscle during morphogenesis of the mammalian diaphragm
EP2730585B1 (fr) Complexe de protéine-peptide provenant de tissus d'embryons d'ongulés, utilisation sur la peau, procédé de production et composition pharmaceutique le comprenant
Ma et al. Recombinant human erythropoietin protects myocardial cells from apoptosis via the janus-activated kinase 2/signal transducer and activator of transcription 5 pathway in rats with epilepsy
Wang et al. Receptor-interacting protein 3/calmodulin-dependent kinase II/proline-rich tyrosine kinase 2 pathway is involved in programmed cell death in a mouse model of brain ischaemic stroke
KR102242040B1 (ko) 줄기세포의 티오레독신 발현을 증진시키는 방법
WO2014051338A2 (fr) Composition pharmaceutique destinée à être utilisée pour la prévention ou le traitement, comprenant une cellule mononuclée de sang périphérique comme principe actif
CN110051842A (zh) 诱导肿瘤细胞转变为神经元样细胞以抑制肿瘤生长的制剂
Zhang et al. HucMSC-EVs facilitate in vitro development of maternally aged preantral follicles and oocytes
WO2021096089A1 (fr) Nouvelle composition comprenant des exosomes dérivés de cellules souches et du polydésoxyribonucléotide en tant que principes actifs
WO2010077093A2 (fr) Composition favorisant la croissance des cheveux ou empêchant la perte de cheveux comprenant de l'érythropoïétine

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19760005

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19760005

Country of ref document: EP

Kind code of ref document: A1