CN113521078A - Aged cell killing agent and application thereof - Google Patents
Aged cell killing agent and application thereof Download PDFInfo
- Publication number
- CN113521078A CN113521078A CN202010318152.1A CN202010318152A CN113521078A CN 113521078 A CN113521078 A CN 113521078A CN 202010318152 A CN202010318152 A CN 202010318152A CN 113521078 A CN113521078 A CN 113521078A
- Authority
- CN
- China
- Prior art keywords
- apilimod
- aging
- cell
- aged
- cells
- 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
Links
- 239000003139 biocide Substances 0.000 title claims abstract description 22
- 230000022534 cell killing Effects 0.000 title claims abstract description 22
- 229950002889 apilimod Drugs 0.000 claims abstract description 57
- HSKAZIJJKRAJAV-KOEQRZSOSA-N n-[(e)-(3-methylphenyl)methylideneamino]-6-morpholin-4-yl-2-(2-pyridin-2-ylethoxy)pyrimidin-4-amine Chemical compound CC1=CC=CC(\C=N\NC=2N=C(OCCC=3N=CC=CC=3)N=C(C=2)N2CCOCC2)=C1 HSKAZIJJKRAJAV-KOEQRZSOSA-N 0.000 claims abstract description 57
- 239000003814 drug Substances 0.000 claims abstract description 31
- 230000003712 anti-aging effect Effects 0.000 claims abstract description 13
- 150000002148 esters Chemical class 0.000 claims abstract description 9
- 150000003839 salts Chemical class 0.000 claims abstract description 5
- 239000012453 solvate Substances 0.000 claims abstract description 5
- 230000032683 aging Effects 0.000 claims description 32
- 238000011282 treatment Methods 0.000 claims description 21
- 239000002552 dosage form Substances 0.000 claims description 6
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 5
- 239000002775 capsule Substances 0.000 claims description 4
- 239000000796 flavoring agent Substances 0.000 claims description 4
- 235000013355 food flavoring agent Nutrition 0.000 claims description 4
- 239000002671 adjuvant Substances 0.000 claims description 3
- 239000011230 binding agent Substances 0.000 claims description 3
- 239000003085 diluting agent Substances 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000000412 dendrimer Substances 0.000 claims description 2
- 229920000736 dendritic polymer Polymers 0.000 claims description 2
- 239000000945 filler Substances 0.000 claims description 2
- 239000010408 film Substances 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000002502 liposome Substances 0.000 claims description 2
- 239000000693 micelle Substances 0.000 claims description 2
- 239000003094 microcapsule Substances 0.000 claims description 2
- 239000004005 microsphere Substances 0.000 claims description 2
- 239000007923 nasal drop Substances 0.000 claims description 2
- 229940100662 nasal drops Drugs 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000000829 suppository Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 230000002147 killing effect Effects 0.000 abstract description 13
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 63
- 210000004027 cell Anatomy 0.000 description 57
- 239000001963 growth medium Substances 0.000 description 23
- 210000002950 fibroblast Anatomy 0.000 description 18
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 15
- 229960005420 etoposide Drugs 0.000 description 15
- 238000005406 washing Methods 0.000 description 12
- 229940079593 drug Drugs 0.000 description 11
- 239000000203 mixture Substances 0.000 description 10
- 210000002966 serum Anatomy 0.000 description 10
- 244000309466 calf Species 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 9
- 230000032677 cell aging Effects 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 238000004113 cell culture Methods 0.000 description 7
- 230000007170 pathology Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000005138 cryopreservation Methods 0.000 description 6
- -1 hydroxytyrosol carbonate compound Chemical class 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 6
- 239000002122 magnetic nanoparticle Substances 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 239000002246 antineoplastic agent Substances 0.000 description 4
- 229940095066 hydroxytyrosol Drugs 0.000 description 4
- JUUBCHWRXWPFFH-UHFFFAOYSA-N hydroxytyrosol Natural products OCCC1=CC=C(O)C(O)=C1 JUUBCHWRXWPFFH-UHFFFAOYSA-N 0.000 description 4
- 235000003248 hydroxytyrosol Nutrition 0.000 description 4
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 3
- 238000000116 DAPI staining Methods 0.000 description 3
- 229930040373 Paraformaldehyde Natural products 0.000 description 3
- 229920004890 Triton X-100 Polymers 0.000 description 3
- 239000013504 Triton X-100 Substances 0.000 description 3
- 229940041181 antineoplastic drug Drugs 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 238000003125 immunofluorescent labeling Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 229920002866 paraformaldehyde Polymers 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 230000003068 static effect Effects 0.000 description 3
- 102000001556 1-Phosphatidylinositol 4-Kinase Human genes 0.000 description 2
- 108010029190 1-Phosphatidylinositol 4-Kinase Proteins 0.000 description 2
- 102100038028 1-phosphatidylinositol 3-phosphate 5-kinase Human genes 0.000 description 2
- 101710145421 1-phosphatidylinositol 3-phosphate 5-kinase Proteins 0.000 description 2
- YCCILVSKPBXVIP-UHFFFAOYSA-N 2-(4-hydroxyphenyl)ethanol Chemical compound OCCC1=CC=C(O)C=C1 YCCILVSKPBXVIP-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 208000011231 Crohn disease Diseases 0.000 description 2
- 230000005778 DNA damage Effects 0.000 description 2
- 231100000277 DNA damage Toxicity 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 239000007884 disintegrant Substances 0.000 description 2
- 230000008030 elimination Effects 0.000 description 2
- 238000003379 elimination reaction Methods 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 206010039073 rheumatoid arthritis Diseases 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229940126585 therapeutic drug Drugs 0.000 description 2
- 239000005660 Abamectin Substances 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 229920002785 Croscarmellose sodium Polymers 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 208000025966 Neurological disease Diseases 0.000 description 1
- 101710138657 Neurotoxin Proteins 0.000 description 1
- 108010090931 Proto-Oncogene Proteins c-bcl-2 Proteins 0.000 description 1
- 102000013535 Proto-Oncogene Proteins c-bcl-2 Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 102000001742 Tumor Suppressor Proteins Human genes 0.000 description 1
- 108010040002 Tumor Suppressor Proteins Proteins 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 239000003560 cancer drug Substances 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 229940097217 cardiac glycoside Drugs 0.000 description 1
- 239000002368 cardiac glycoside Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 229960001681 croscarmellose sodium Drugs 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000007902 hard capsule Substances 0.000 description 1
- 239000003481 heat shock protein 90 inhibitor Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000002581 neurotoxin Substances 0.000 description 1
- 231100000618 neurotoxin Toxicity 0.000 description 1
- 230000005853 oncogenic activation Effects 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 229940125381 senolytic agent Drugs 0.000 description 1
- 230000009327 senolytic effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229930002534 steroid glycoside Natural products 0.000 description 1
- 150000008143 steroidal glycosides Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 108091035539 telomere Proteins 0.000 description 1
- 102000055501 telomere Human genes 0.000 description 1
- 210000003411 telomere Anatomy 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Images
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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to an aged cell killing agent and application thereof, wherein the aged cell killing agent comprises apilimod or pharmaceutically acceptable salts, esters and solvates thereof. The invention discovers for the first time that the medicine Apilimod (Apilimod) has specific killing effect on aged cells, and the killing rate is more than 70 percent, so the medicine Apilimod can be used as a novel aged cell killing agent, and is more likely to be used as a potential novel anti-aging medicine because the medicine Apilimod can effectively eliminate the aged cells.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to an aged cell killing agent and application thereof.
Background
Cell aging (Senescence) is an irreversible growth arrest state induced by a variety of factors. Cell aging occurs in response to various cellular stressors, such as telomere shortening, DNA damage, oxidative stress, and oncogenic activation. Cellular aging may play an important role in wound healing and prevention of tissue fibrosis, and is more a broad mechanism of tumor inhibition. In cancer Therapy using compounds, the phenomenon of cancer drugs inducing cell aging is referred to as Therapy-Induced cell aging (Therapy-Induced Cellular science). Recent studies have shown that TIS can serve as a new tumor suppressor target, inducing tumor cell aging after targeting by specific anti-tumor drugs or radiation therapy, and thus this process can serve as a new tumor treatment strategy.
Etoposide (ETO) is a topoisomerase inhibitor that induces cellular aging by causing DNA damage and is therefore commonly used as a chemotherapeutic agent to treat various types of cancer. The early-stage study on Apilimod (Apilimod) finds that the Apilimod is an inhibitor of IL-2/IL-23 and is a potential therapeutic drug for Crohn's disease and rheumatoid arthritis; however, recent studies show that Apilimod is also an inhibitor of phosphoinositide kinase PIKfyve and is a potential target for cancer treatment, and Apilimod is currently used as an anti-cancer drug for clinical research.
Cellular aging is a complex process associated with many human aging-related disorders, such as cardiovascular disease, type ii diabetes, osteoarthritis, and neurological disorders; the mouse model of aging proves that the inhibition of the gene expression of aging signal pathway and the elimination of aging cells are helpful for the improvement of aging diseases of the mouse model, so the elimination of relevant aging cells in the clinical treatment of human aging diseases is also one of important research directions. In recent years, an emerging strategy has been to specifically eliminate aging cells using compounds known as anti-aging (senolytics). Several classes of anti-aging compounds have been found to include cardiac glycosides, inhibitors of the BCL-2 protein family, avermectins (HSP90 inhibitors), etc., and anti-aging compounds have now entered the clinical validation phase.
CN105916835A also discloses a lipophilic hydroxytyrosol carbonate compound, which can be prepared by oxidizing a substituted hydroxybenzaldehyde compound with an oxidizing agent such as hydrogen peroxide under mild conditions. The method comprises enzymatically esterifying 4- (2-hydroxyethyl) phenol to form a corresponding ester, introducing a formyl group ortho to the phenolic hydroxyl group of the ester to form a lipophilic formyltyrosol ester; oxidizing the lipophilic formyltyrosol ester with a peroxide compound to form a lipophilic hydroxytyrosol ester compound; and reacting the lipophilic hydroxytyrosol ester compound with a carbonate derivative to form the lipophilic hydroxytyrosol carbonate compound. These compounds can be used as anti-aging compounds in a wide variety of personal care compositions that are stable to oxidation.
CN106176809A also discloses a Fe3O4Application of magnetic nanoparticles in preparing anti-aging agent is provided. The anti-aging agent is Fe3O4The magnetic nanoparticles are active ingredients and are Fe coated with edible nutrient substances3O4Magnetic nanoparticles, edible nutrient coated Fe3O4The magnetic nano-particles have higher water phase stability and biocompatibility, and can lead Fe3O4Magnetic nanoparticles enter cells; fe introduced into the cells3O4The magnetic nanoparticles can remarkably reduce the increase of intracellular ROS level and apoptosis induced by hydrogen peroxide or neurotoxin in a certain dosage range, eliminate excessive oxygen free radicals in cells, and relieve oxidative stress caused by external stimulation, aging or gene dysfunction, thereby having the effect of resisting aging.
Because of the complexity of the aging process, different anti-aging compounds inhibit a certain pathway, and thus the development of different anti-aging compounds is important.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide an aged cell killing agent and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
in one aspect, the present invention provides an aging cell killing agent comprising apilimod or a pharmaceutically acceptable salt, ester, solvate thereof.
The aging cell killing agent according to the present invention may include only apilimod, any one of apilimod pharmaceutically acceptable salt, apilimod pharmaceutically acceptable ester or apilimod pharmaceutically acceptable solvate, or a combination of at least two of apilimod pharmaceutically acceptable salt, apilimod pharmaceutically acceptable ester, apilimod pharmaceutically acceptable solvate, and the like, and any combination may be selected, which is not repeated herein.
Early research on the drug Apilimod (Apilimod) finds that the drug Apilimod is an inhibitor of IL-2/IL-23 and is a potential therapeutic drug for Crohn's disease and rheumatoid arthritis; recent studies indicate that Apilimod is also an inhibitor of phosphoinositide kinase PIKfyve and is a potential target for cancer treatment, and Apilimod is currently used in clinical research as an anti-cancer drug.
The invention discovers for the first time that the medicine Apilimod (Apilimod) has specific killing effect on aged cells including treatment-induced aged cells or natural aging, the killing rate is more than 70 percent, and the medicine has no damage to non-aged cells, so the medicine can be used as a novel aged cell killing agent, and can be more likely to be used as a potential novel anti-aging medicine because the medicine can efficiently remove the aged cells.
Preferably, the aging cells are therapy-induced aging cells and/or naturally aging cells.
According to the invention, aged cells formed by human fibroblasts induced by drug etoposide and aged human fibroblasts in a natural state are used as research models, and researches show that Apilimod (Apilimod) has a high killing effect on the aged cells formed by human fibroblasts induced by etoposide and the aged human fibroblasts in the natural state, and has no killing effect on non-aged cells.
Preferably, the dosage form of the aged cell killing agent comprises tablets, suspensions, disintegrants, capsules, granules, powders, emulsions, injections, sprays, films, suppositories, nasal drops or dropping pills.
Preferably, the aged cell killing agent further comprises a pharmaceutically acceptable adjuvant.
Preferably, the excipient comprises any one of, or a combination of at least two of, an excipient, a diluent, a carrier, a flavoring agent, a binder, or a filler. The combination of the at least two compounds, such as the combination of excipient and carrier, the combination of diluent and carrier, the combination of flavoring agent and binder, etc., can be selected in any combination manner, and will not be described in detail herein.
Preferably, the carrier comprises a liposome, micelle, dendrimer, microsphere or microcapsule.
The apilimod serving as the aged cell killing agent can be independently administered or can be matched with auxiliary materials to be prepared into a proper dosage form for administration, and when the dosage form is a tablet, the apilimod can contain excipients, such as microcrystalline cellulose, starch or calcium carbonate and the like; disintegrants may also be included, such as croscarmellose sodium and the like; when the dosage form is a capsule, the preparation can be prepared into a hard capsule or a soft capsule, and the apilimod and the auxiliary materials can be prepared into powder or granules to be filled into the capsule; when the preparation is suspension, flavoring agent, suspending agent, etc. can be added to adjust taste and mouthfeel. When the dosage form is emulsion, emulsifier and cosolvent can be added to adjust solubility and emulsifying degree for administration.
The apilimod serving as the aging cell killing agent can be matched with other medicines with the effect of killing aging cells or inhibiting the activity of the aging cells according to different proportions to form a medicine composition, and the medicine composition plays a role together.
In another aspect, the present invention provides a use of the above-mentioned aging cell killer for preparing an anti-aging drug.
Compared with the prior art, the invention has the following beneficial effects:
the invention discovers for the first time that the medicine Apilimod (Apilimod) has specific killing effect on aged cells including treatment-induced aged cells or natural aging, the killing rate is more than 70 percent, and the medicine has no damage to non-aged cells, so the medicine can be used as a novel aged cell killing agent, and can be more likely to be used as a potential novel anti-aging medicine because the medicine can efficiently remove the aged cells.
Drawings
Fig. 1 is a graph of the imaging of pathology for the apilimod-treated group and the DMSO-treated group in example 1 (a is the apilimod-treated group, b is the DMSO-treated group);
fig. 2 is a graph of the imaging of pathology for the apilimod-treated group and the DMSO-treated group in example 2 (a is the apilimod-treated group, b is the DMSO-treated group);
FIG. 3 is a graph of imaging pathology in the apilimod-treated group of example 3;
fig. 4 is an image of pathology for the DMSO-treated group in example 3.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
The following examples relate to complete media formulations: 89% DMEM (Dulbeccos Modified Eagles Medium, High Glucose, GE Healthcare Cell Culture) + 10% FBS (Total bone serum, NTC, SFBE) + 1% PS (penillin: Streptomyces solution, Hyclone)TM)。
The drug Apilimod (Apilimod) referred to in the following examples was purchased from: a ceramic biomass.
The drug Etoposide (Etoposide, ETO) referred to in the following examples was purchased from: abcam.
Wherein the human fibroblasts used in example 1 and example 2 (IMR-90:)
CCL-186TM) Cells at passage 15), cells were in a non-aged state (IMR 90P 15); human fibroblast used in example 3 (IMR-90: (
CCL-186TM) Cells were in a state of natural aging for 27 passages (IMR 90P 27).
Example 1
Step 1: the recovery of human fibroblasts (IMR 90P 15) was performed as follows:
(1) taking out the human fibroblast cryopreservation tube from the liquid nitrogen tank, and carrying out water bath at 37 ℃ until the human fibroblast cryopreservation tube is melted;
(2) opening the cover of the freezing tube, and adding a proper amount of complete culture medium;
(3) centrifuging at 1000rpm for 5 min;
(4) discarding supernatant, adding culture medium containing 10% calf serum, resuspending cells, inoculating to 10cm cell culture dish, and static culturing at 37 deg.C in incubator (0.5% CO)2，0.3％O2，RH 99％)；
(5) The culture medium was changed the next day and the cultivation was continued for 24 h.
Step 2: for human fibroblast (IMR-90: (
CCL-186TM) To perform resuscitation, the operation is as follows:
(1) removing the cells from the incubator, removing the medium using a pipettor, and washing for 3min with 10mL PBS;
(2) removing PBS, adding 1mL of trypsin-EDTA (0.25%), slowly shaking, mixing, and digesting at 37 deg.C for 3 min;
(3) adding 2mL culture medium containing 10% calf serum, and stopping digestion;
(4) collecting the mixture in a centrifuge tube, and centrifuging the mixture for 5min at 1000 rpm;
(5) removing supernatant, adding culture medium containing 10% calf serum, resuspending cells, counting cells, adjusting cell density, and inoculating to 96-well cell culture plate to make the number of cells in each well be 0.7 ten thousand;
(6) etoposide (Etoposide) was added to the cell culture medium used for plating so that the final concentration of Etoposide was 1 μ M.
And step 3: adding drugs for treatment, and setting a DMSO treatment group and an apilimod treatment group, wherein the operations are as follows:
(1) after etoposide treatment for 48h, dosing treatment is carried out, a DMSO treatment group is added into a complete culture medium containing 1 mu M etoposide and 1 mu M DMSO, and an apilimod treatment group is added into a complete culture medium containing 1 mu M etoposide, 1 mu M apilimod and 1 mu M DMSO;
(2) after the medicine is added for 48 hours, the complete culture medium is replaced to continue the culture for 24 hours.
And 4, step 4: immunofluorescent staining and cell counting, performed as follows:
(1) removing the culture medium, washing with PBS for 3 times, each for 5 min;
(2) fixing: fixing 4% paraformaldehyde at 25 deg.C for 15min, and washing with PBS for 5min for 3 times;
(3) permeability: PBS containing 0.5% Triton X-100 is transparent for 15min at 25 deg.C, and washed with PBS for 3 times, 5min each time;
(4) DAPI staining: diluting 5mg/mL DAPI with PBS according to the proportion of 1:1000, and incubating for 1h in a dark place with 100 mu L of each well;
(5) washing with PBS for 5min for 3 times;
(6) after the staining was completed, a photograph was taken using a full-automatic high-throughput high-content pathology imaging system (ImageXpress Micro consistent), as shown in fig. 1 (a is apilimod-treated group, b is DMSO-treated group);
(7) after the photographing is finished, the Cell counting is carried out by using the Multi wavelet Cell screening function built in the software, each sample is parallelly detected for 10 times, and the data result is presented as an average value. The results are shown in Table 1.
Example 2
Step 1: the recovery of human fibroblasts (IMR 90P 15) was performed as follows:
(1) taking out the human fibroblast cryopreservation tube from the liquid nitrogen tank, and carrying out water bath at 37 ℃ until the human fibroblast cryopreservation tube is melted;
(2) opening the cover of the freezing tube, and adding a proper amount of complete culture medium;
(3) centrifuging at 1000rpm for 5 min;
(4) discarding supernatant, adding culture medium containing 10% calf serum, resuspending cells, inoculating to 10cm cell culture dish, and static culturing at 37 deg.C in incubator (0.5% CO)2，0.3％O2，RH 99％)；
(5) The culture medium was changed the next day and the cultivation was continued for 24 h.
Step 2: for human fibroblast (IMR-90: (
CCL-186TM) To perform resuscitation, the operation is as follows:
(1) removing the cells from the incubator, removing the medium using a pipettor, and washing for 3min with 10mL PBS;
(2) removing PBS, adding 1mL of trypsin-EDTA (0.25%), slowly shaking, mixing, and digesting at 37 deg.C for 3 min;
(3) adding 2mL culture medium containing 10% calf serum, and stopping digestion;
(4) collecting the mixture in a centrifuge tube, and centrifuging the mixture for 5min at 1000 rpm;
(5) removing supernatant, adding culture medium containing 10% calf serum, resuspending cells, counting cells, adjusting cell density, and inoculating to 96-well cell culture plate to make the number of cells per well be 0.7 ten thousand.
And step 3: adding drugs for treatment, and setting a DMSO treatment group and an apilimod treatment group, wherein the operations are as follows:
(1) the DMSO-treated group was added to complete medium containing 1 μ M DMSO, the apilimod-treated group was added to complete medium containing 1 μ M apilimod and 1 μ M DMSO;
(2) after the medicine is added for 48 hours, the complete culture medium is replaced to continue the culture for 24 hours.
And 4, step 4: immunofluorescent staining and cell counting, performed as follows:
(1) removing the culture medium, washing with PBS for 3 times, each for 5 min;
(2) fixing: fixing 4% paraformaldehyde at 25 deg.C for 15min, and washing with PBS for 5min for 3 times;
(3) permeability: PBS containing 0.5% Triton X-100 is transparent for 15min at 25 deg.C, and washed with PBS for 3 times, 5min each time;
(4) DAPI staining: diluting 5mg/mL DAPI with PBS according to the proportion of 1:1000, and incubating for 1h in a dark place with 100 mu L of each well;
(5) washing with PBS for 5min for 3 times;
(6) after the staining was completed, a photograph was taken using a full-automatic high-throughput high-content pathology imaging system (ImageXpress Micro consistent), as shown in fig. 2 (a is apilimod-treated group, b is DMSO-treated group);
(7) after the photographing is finished, the Cell counting is carried out by using the Multi wavelet Cell screening function built in the software, each sample is parallelly detected for 10 times, and the data result is presented as an average value. The results are shown in Table 1.
Example 3
Step 1: the recovery of human fibroblasts (IMR 90P 27) was performed as follows:
(1) taking out the human fibroblast cryopreservation tube from the liquid nitrogen tank, and carrying out water bath at 37 ℃ until the human fibroblast cryopreservation tube is melted;
(2) opening the cover of the freezing tube, and adding a proper amount of complete culture medium;
(3) centrifuging at 1000rpm for 5 min;
(4) discarding supernatant, adding culture medium containing 10% calf serum, resuspending cells, inoculating to 10cm cell culture dish, and static culturing at 37 deg.C in incubator (0.5% CO)2，0.3％O2，RH 99％)；
(5) The culture medium was changed the next day and the cultivation was continued for 24 h.
Step 2: for human fibroblast (IMR-90: (
CCL-186TM) To perform resuscitation, the operation is as follows:
(1) removing the cells from the incubator, removing the medium using a pipettor, and washing for 3min with 10mL PBS;
(2) removing PBS, adding 1mL of trypsin-EDTA (0.25%), slowly shaking, mixing, and digesting at 37 deg.C for 3 min;
(3) adding 2mL culture medium containing 10% calf serum, and stopping digestion;
(4) collecting the mixture in a centrifuge tube, and centrifuging the mixture for 5min at 1000 rpm;
(5) removing supernatant, adding culture medium containing 10% calf serum, resuspending cells, counting cells, adjusting cell density, and inoculating to 96-well cell culture plate to make the number of cells per well be 0.7 ten thousand.
And step 3: adding drugs for treatment, and setting a DMSO treatment group and an apilimod treatment group, wherein the operations are as follows:
(1) the DMSO-treated group was added to complete medium containing 1 μ M DMSO, the apilimod-treated group was added to complete medium containing 1 μ M apilimod and 1 μ M DMSO;
(2) after the medicine is added for 48 hours, the complete culture medium is replaced to continue the culture for 24 hours.
And 4, step 4: immunofluorescent staining and cell counting, performed as follows:
(1) removing the culture medium, washing with PBS for 3 times, each for 5 min;
(2) fixing: fixing 4% paraformaldehyde at 25 deg.C for 15min, and washing with PBS for 5min for 3 times;
(3) permeability: PBS containing 0.5% Triton X-100 is transparent for 15min at 25 deg.C, and washed with PBS for 3 times, 5min each time;
(4) DAPI staining: diluting 5mg/mL DAPI with PBS according to the proportion of 1:1000, and incubating for 1h in a dark place with 100 mu L of each well;
(5) washing with PBS for 5min for 3 times;
(6) after staining was completed, photographs were taken using a full-automatic high-throughput high-content pathology imaging system (ImageXpress Micro consistent), as shown in fig. 3 (apilimod-treated group) and fig. 4 (DMSO-treated group);
(7) after the photographing is finished, the Cell counting is carried out by using the Multi wavelet Cell screening function built in the software, each sample is parallelly detected for 10 times, and the data result is presented as an average value. The results are shown in Table 1.
TABLE 1
| Group of | State of the cell | Apilimod treatment group | DMSO treatment group | Rate of killing |
| Example 1 | P15+ETO | 349 | 1369 | 74.5％ |
| Example 2 | P15 | 21578 | 23850 | 9.53％ |
| Example 3 | P27 | 331 | 2070 | 84.0％ |
As can be seen from the data in Table 1: apilimod (Apilimod) has obvious specific killing effect on aged cells formed by induction of etoposide or aged cells in a natural state (examples 1 and 3), the killing rate is over 70 percent, and almost no killing effect is generated on non-aged cells (example 2), and the Apilimod can be used as a novel aged cell killing agent to efficiently remove the aged cells.
The applicant states that the present invention is illustrated by the above examples to show an aged cell killing agent and its application, but the present invention is not limited to the above examples, i.e., it does not mean that the present invention must be implemented by the above examples. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
Claims (7)
1. An aging cell killing agent, wherein the aging cell killing agent comprises apilimod or pharmaceutically acceptable salts, esters and solvates thereof.
2. The aging cell killer of claim 1, wherein the aging cell is a treatment-induced aging cell and/or a natural aging cell.
3. The senescent cell killing agent according to claim 1 or 2, wherein the dosage form of the senescent cell killing agent comprises tablets, capsules, granules, powder, injections, sprays, films, suppositories, nasal drops or dropping pills.
4. The aging cell killer of any one of claims 1 to 3, further comprising a pharmaceutically acceptable adjuvant.
5. The aged cell killing agent of claim 4, wherein the adjuvant comprises any one or a combination of at least two of an excipient, a diluent, a carrier, a flavoring agent, a binder, or a filler.
6. The aging cell killer of claim 5, wherein the carrier comprises a liposome, a micelle, a dendrimer, a microsphere, or a microcapsule.
7. Use of the aging cell killer of any one of claims 1 to 6 for the preparation of an anti-aging medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010318152.1A CN113521078A (en) | 2020-04-21 | 2020-04-21 | Aged cell killing agent and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010318152.1A CN113521078A (en) | 2020-04-21 | 2020-04-21 | Aged cell killing agent and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113521078A true CN113521078A (en) | 2021-10-22 |
Family
ID=78123841
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010318152.1A Pending CN113521078A (en) | 2020-04-21 | 2020-04-21 | Aged cell killing agent and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113521078A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023147605A3 (en) * | 2022-01-31 | 2023-10-19 | Sens Research Foundation | Senescent cell surface markers |
-
2020
- 2020-04-21 CN CN202010318152.1A patent/CN113521078A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023147605A3 (en) * | 2022-01-31 | 2023-10-19 | Sens Research Foundation | Senescent cell surface markers |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102239149B (en) | Quinoline compounds as inhibitors of angiogenesis, human methionine aminopeptidase, and sirt1, and methods of treating disorders | |
| CN106456658A (en) | Compositions of selenoorganic compounds and methods of use thereof | |
| CN104109145A (en) | Flavonoid structure based hydrogen sulfide donor derivative and application in treatment of neuroinflammation related diseases thereof | |
| CN102552908A (en) | Pharmaceutical composition containing artemisinin, artemisinin derivatives and Bcl-2 inhibitor and application thereof | |
| Xing et al. | Astragalin mitigates inflammatory osteolysis by negatively modulating osteoclastogenesis via ROS and MAPK signaling pathway | |
| CN113521078A (en) | Aged cell killing agent and application thereof | |
| CN102441168B (en) | Medicine composition containing apigenin, apigenin derivant and Bc1-2 inhibitor and application thereof in preparation of medicines capable of treating cancer | |
| CN101940569B (en) | Medicament composition containing sorafenib, artemisinin and artemisinin derivative and application thereof in preparing medicament for treating cancer | |
| CN106748939B (en) | A kind of novel bromine phenol thiosemicarbazide compound and its preparation and drug and purposes | |
| CN102688493A (en) | Pharmaceutical composition containing resveratrol, resveratrol derivative and Bc1-2 inhibitor, and application thereof | |
| CN102688489B (en) | Pharmaceutical composition containing triptolide, triptolide derivative and Bc1-2 inhibitor and application thereof | |
| CN101461819A (en) | Use of mangiferin calcium salt as peroxisome proliferator-activated receptor agonist | |
| CN105193795A (en) | Application of two halogen-phenol compounds to effect of promoting angiogenesis | |
| CN114522168A (en) | Pharmaceutical composition for treating gastric cancer and application | |
| CN110575450A (en) | Application of 2, 5-furandimethanol in preparation of antitumor drugs | |
| CN112972455B (en) | Application of compound in preparation of antitumor drugs | |
| CN104926804A (en) | Compounds with anti-tumor effect, and preparation method and application of compounds | |
| CN110876745A (en) | Resveratrol and procyanidin composition with high anticancer activity | |
| CN110063988A (en) | A kind of pharmaceutical composition and preparation method thereof for treating neuroblastoma | |
| CN104208074B (en) | Purposes of the 2 α hydroxyl protopanoxadiols in tumor multi-drug resistance reversal agents are prepared | |
| CN103880793B (en) | Containing furan imine compound and its production and use | |
| CN103833766B (en) | Shore, Agra dimethylamine fumarate and the purposes in prepared by medicine thereof | |
| CN102688490A (en) | Pharmaceutical composition containing evodiamine, evodiamine derivative and Bc1-2 inhibitor, and the application | |
| CN101653607A (en) | Pharmaceutical composition containing hepatocyte growth factor receptor inhibitor and mitogen extracellular kinase inhibitor and application thereof | |
| CN106852926A (en) | Application of the triterpene glucoside unit in antineoplastic sensitizer is prepared |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20211022 |
|
| WD01 | Invention patent application deemed withdrawn after publication |