HRP20150510T1 - Äśestice mezenhimskih matiäśnih stanica - Google Patents
Äśestice mezenhimskih matiäśnih stanica Download PDFInfo
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- HRP20150510T1 HRP20150510T1 HRP20150510TT HRP20150510T HRP20150510T1 HR P20150510 T1 HRP20150510 T1 HR P20150510T1 HR P20150510T T HRP20150510T T HR P20150510TT HR P20150510 T HRP20150510 T HR P20150510T HR P20150510 T1 HRP20150510 T1 HR P20150510T1
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- Croatia
- Prior art keywords
- exosome
- disease
- mesenchymal stem
- kda
- complexes
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- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims 8
- 239000002245 particle Substances 0.000 title 1
- 210000001808 exosome Anatomy 0.000 claims 23
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 5
- 108090000623 proteins and genes Proteins 0.000 claims 5
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims 4
- 201000010099 disease Diseases 0.000 claims 4
- 102000004169 proteins and genes Human genes 0.000 claims 4
- 229920000858 Cyclodextrin Polymers 0.000 claims 3
- 230000004071 biological effect Effects 0.000 claims 3
- 239000008194 pharmaceutical composition Substances 0.000 claims 3
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 claims 3
- 208000027418 Wounds and injury Diseases 0.000 claims 2
- 235000012000 cholesterol Nutrition 0.000 claims 2
- 230000006378 damage Effects 0.000 claims 2
- 239000003599 detergent Substances 0.000 claims 2
- 208000014674 injury Diseases 0.000 claims 2
- 150000002632 lipids Chemical class 0.000 claims 2
- 238000000034 method Methods 0.000 claims 2
- 150000003904 phospholipids Chemical class 0.000 claims 2
- 229920002477 rna polymer Polymers 0.000 claims 2
- JSPNNZKWADNWHI-PNANGNLXSA-N (2r)-2-hydroxy-n-[(2s,3r,4e,8e)-3-hydroxy-9-methyl-1-[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoctadeca-4,8-dien-2-yl]heptadecanamide Chemical compound CCCCCCCCCCCCCCC[C@@H](O)C(=O)N[C@H]([C@H](O)\C=C\CC\C=C(/C)CCCCCCCCC)CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O JSPNNZKWADNWHI-PNANGNLXSA-N 0.000 claims 1
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 claims 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 claims 1
- 208000024827 Alzheimer disease Diseases 0.000 claims 1
- 206010059313 Anogenital warts Diseases 0.000 claims 1
- 208000009137 Behcet syndrome Diseases 0.000 claims 1
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 claims 1
- 201000003883 Cystic fibrosis Diseases 0.000 claims 1
- 102100025621 Cytochrome b-245 heavy chain Human genes 0.000 claims 1
- 201000004624 Dermatitis Diseases 0.000 claims 1
- 206010012438 Dermatitis atopic Diseases 0.000 claims 1
- 101150082674 E2 gene Proteins 0.000 claims 1
- 206010016654 Fibrosis Diseases 0.000 claims 1
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 claims 1
- 229930186217 Glycolipid Natural products 0.000 claims 1
- 206010019280 Heart failures Diseases 0.000 claims 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims 1
- 206010061216 Infarction Diseases 0.000 claims 1
- 206010061218 Inflammation Diseases 0.000 claims 1
- 208000002260 Keloid Diseases 0.000 claims 1
- 206010023330 Keloid scar Diseases 0.000 claims 1
- 208000007201 Myocardial reperfusion injury Diseases 0.000 claims 1
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 claims 1
- 208000018737 Parkinson disease Diseases 0.000 claims 1
- 201000004681 Psoriasis Diseases 0.000 claims 1
- 208000000453 Skin Neoplasms Diseases 0.000 claims 1
- 208000031673 T-Cell Cutaneous Lymphoma Diseases 0.000 claims 1
- 101150052863 THY1 gene Proteins 0.000 claims 1
- 229920004890 Triton X-100 Polymers 0.000 claims 1
- 208000025865 Ulcer Diseases 0.000 claims 1
- 241000934136 Verruca Species 0.000 claims 1
- 208000000260 Warts Diseases 0.000 claims 1
- 238000002835 absorbance Methods 0.000 claims 1
- 238000004458 analytical method Methods 0.000 claims 1
- 239000000427 antigen Substances 0.000 claims 1
- 102000036639 antigens Human genes 0.000 claims 1
- 108091007433 antigens Proteins 0.000 claims 1
- 101150010487 are gene Proteins 0.000 claims 1
- 201000008937 atopic dermatitis Diseases 0.000 claims 1
- 208000015322 bone marrow disease Diseases 0.000 claims 1
- 201000011510 cancer Diseases 0.000 claims 1
- 230000005961 cardioprotection Effects 0.000 claims 1
- 230000030833 cell death Effects 0.000 claims 1
- 229940106189 ceramide Drugs 0.000 claims 1
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 claims 1
- 229930183167 cerebroside Natural products 0.000 claims 1
- RIZIAUKTHDLMQX-UHFFFAOYSA-N cerebroside D Natural products CCCCCCCCCCCCCCCCC(O)C(=O)NC(C(O)C=CCCC=C(C)CCCCCCCCC)COC1OC(CO)C(O)C(O)C1O RIZIAUKTHDLMQX-UHFFFAOYSA-N 0.000 claims 1
- 230000001684 chronic effect Effects 0.000 claims 1
- 208000016532 chronic granulomatous disease Diseases 0.000 claims 1
- 201000007241 cutaneous T cell lymphoma Diseases 0.000 claims 1
- 206010012601 diabetes mellitus Diseases 0.000 claims 1
- 208000035475 disorder Diseases 0.000 claims 1
- 238000002296 dynamic light scattering Methods 0.000 claims 1
- 230000008482 dysregulation Effects 0.000 claims 1
- 238000001493 electron microscopy Methods 0.000 claims 1
- 230000004761 fibrosis Effects 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 201000011066 hemangioma Diseases 0.000 claims 1
- 230000028993 immune response Effects 0.000 claims 1
- 238000000338 in vitro Methods 0.000 claims 1
- 230000007574 infarction Effects 0.000 claims 1
- 230000004054 inflammatory process Effects 0.000 claims 1
- 208000037906 ischaemic injury Diseases 0.000 claims 1
- 210000001117 keloid Anatomy 0.000 claims 1
- 230000003902 lesion Effects 0.000 claims 1
- 239000012528 membrane Substances 0.000 claims 1
- 108091070501 miRNA Proteins 0.000 claims 1
- 239000002679 microRNA Substances 0.000 claims 1
- 238000010172 mouse model Methods 0.000 claims 1
- 201000005962 mycosis fungoides Diseases 0.000 claims 1
- 208000010125 myocardial infarction Diseases 0.000 claims 1
- 208000031225 myocardial ischemia Diseases 0.000 claims 1
- 208000015122 neurodegenerative disease Diseases 0.000 claims 1
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 claims 1
- 230000000399 orthopedic effect Effects 0.000 claims 1
- 230000036542 oxidative stress Effects 0.000 claims 1
- 230000001575 pathological effect Effects 0.000 claims 1
- 239000000546 pharmaceutical excipient Substances 0.000 claims 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims 1
- 150000003905 phosphatidylinositols Chemical class 0.000 claims 1
- 238000013310 pig model Methods 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 208000025638 primary cutaneous T-cell non-Hodgkin lymphoma Diseases 0.000 claims 1
- 206010039083 rhinitis Diseases 0.000 claims 1
- 238000000926 separation method Methods 0.000 claims 1
- 201000009890 sinusitis Diseases 0.000 claims 1
- 238000001542 size-exclusion chromatography Methods 0.000 claims 1
- 201000000849 skin cancer Diseases 0.000 claims 1
- 208000017520 skin disease Diseases 0.000 claims 1
- 201000010153 skin papilloma Diseases 0.000 claims 1
- 239000002904 solvent Substances 0.000 claims 1
- 150000003431 steroids Chemical class 0.000 claims 1
- 230000007838 tissue remodeling Effects 0.000 claims 1
- 230000001960 triggered effect Effects 0.000 claims 1
- 230000036269 ulceration Effects 0.000 claims 1
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
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Claims (14)
1. Egzosom izoliran iz mezenhimske matične stanice i s veličinom između 50-100 nm, potvrđenu elektronskom mikroskopijom, s time da egzosom sadrži najmanje jedno biološko svojstvo mezenhimske matične stanice kao što je kod biološke aktivnosti u kontroliranom mediju mezenhimkih matičnih stanica (KM-MMS).
2. Egzosom u skladu sa zahtjevom 1, pri čemu biološka aktivnost sadrži kardioprotekciju.
3. Egzosom u skladu sa zahtjevom 1 ili 2, koji može reducirati veličinu infarkta kao što je primjerice ispitano na mišjem ili svinjskom modelu miokardijalne ishemije ili reperfuzijske ozljede, ili koji može reducirati oksidativni stres kao što je primjerice ispitano in vitro analizom vodikovim peroksidom(H2O2) kod inducirane stanične smrti.
4. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu egzosom sadrži najmanje 70% proteina iz kontroliranog medija mezenhimskih matičnih stanica (KM-MMS), kao što su izabrani iz tablice D1 ili E2, ili su genski produkti gena prikazanih na tablici D2, ili sadrže jedan ili više miRNA kao što je prikazano na tablici E3.
5. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu se egzosom sastoji od:
a) Kompleksa molekulske mase >100 kDa, primjerice sadržavajući proteine <100 kDa;
b) Kompleksa molekulske mase >300 kDa, primjerice sadržavajući proteine <300 kDa;ili
c) Kompleksa molekulske mase >1000 kDa.
6. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu je egzosom veličine između 50 i 100 nm, određeno filtracijom s 0,2 µM filtrom i koncentracijom s membranom granične molekulske mase do 10 kDa, ili hidrodinamičkim radijusom ispod 100nm, kao što je između 30 i 70 nm, 40 i 60 nm, 45 i 55 nm ili oko 50 nm, kao što je određeno primjerice laserskom difrakcijom ili dinamičkim raspršivanjem svjetla.
7. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu egzosom sadži lipid izabran iz skupine koja se sastoji od: fosfolipida, fosfatidil serina, fosfatidil inozitola, fosfatidil kolina, sfingomijelina, ceramida, glikolipida, cerebrozida, steroida, kolesterola, pri čemu je primjerice odnos kolesterola i fosfolipida veći od 0,3-0,4(mol/mol).
8. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu egzosom sadrži lipidni raft, ili pri čemu je egzosom netopljiv u neionskom detergentu, posebice Triton-X100, ili pri čemu je egzosom takav da proteini molekulskih masa definiranih zahtjevom 4 većinski ostanu unutar kompleksa molekulskih masa definiranih tim zahtjevima kada je tretiran s neionskim detergentom, ili je egzosom osjetljiv na ciklodekstrin, poželjno 20 mM ciklodekstrin, tako da tretiranje ciklodekstrinom uzrokuje značajno razdvajanje kompleksa definiranih u zahtjevu 5.
9. Egzosom u skladu s bilo kojim od prethodnih zahtjeva, pri čemu egzosom sadrži ribonukleinsku kiselinu(RNA), pri čemu je poželjno da egzosom ima apsorbanciju u vrijednosti od 1.9 (260:280 nm), ili pri čemu egzosom sadrži površinski antigen izabran iz skupine koja se sastoji od: CD9, CD109 and thy-1.
10. Metoda proizvodnje egzosoma prema bilo kojem od prethodnih zahtjeva, koja sadrži:
a) Dobivanje kontroliranog medija mezenhimskih matičnih stanica (KM-MMS);
b) Koncentriranje kontroliranog medija mezenhimskih matičnih stanica;
c) Stavljanje koncentriranog kontroliranog medija mezenhimskih matičnih stanica na kromatografiju isključenjem prema veličini;
d) Odabir frakcija koje apsorbiraju UV zrake.
11. Farmaceutski pripravak koji sadrži egzosom u skladu s bilo kojim od zahtjeva 1-9 i farmaceutski prihvatljiv ekscipijens, otapalo ili nosač.
12. Egzosom u skladu s bilo kojim od zahtjeva 1-9 ili farmaceutski pripravak u skladu sa zahtjevom 11 za uporabu u metodi za liječenje bolesti.
13. Uporaba egzosoma prema bilo kojem od zahtjeva 1-9 za pripremu farmaceutskog pripravka za liječenje bolesti, pri čemu je bolest izabrana iz skupine koja se sastoji od: srčanog zatajenja, bolesti koštane srži, kožnih bolesti, opeklina i degenerativnih bolesti kao što su dijabetes, Alzheimerova bolest, Parkinsonova bolest, rak, miokardijalni infarkt, kutana ozljeda, dermatološki poremećaj, dermatološka lezija, dermatitis, psorijaza, kondilom, veruka, hemangiom, keloid, rak kože, atopički dermatitis, Behcetova bolest, kronična granulomatozna bolest, kutani T-stanični limfom, ulceracija, patološko stanje određeno s inicijalnom ozljedom koja je potakla upalu i disregulaciju imunološkog odgovora što vodi do kroničnog remodeliranja tkiva koje uključuje fibrozu i gubitak funkcije, renalnu ishemijsku ozljedu, cističnu fibrozu, sinusitis i rinitis ili ortopedsku bolest.
14. Farmaceutski oblik za primjenu egzosoma koji sadrži egzosom, u skladu s bilo kojim od zahtjeva 1-9, zajedno s dozatorom koji ima ulogu dostave egzosoma do mete.
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| PT2254586E (pt) | 2015-05-18 |
| WO2009105044A1 (en) | 2009-08-27 |
| CN104127438A (zh) | 2014-11-05 |
| JP2011513217A (ja) | 2011-04-28 |
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