Esfandiari, Maryam

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    داﻧﺸﮕﺎه ﻋﻠﻮم ﭘﺰﺷﻜﻲ و ﺧﺪﻣﺎت ﺑﻬﺪاﺷﺘﻲ درﻣﺎﻧﻲ ﻛﺮﻣﺎن  داﻧﺸﻜﺪه داروﺳﺎزي و ﻋﻠﻮم داروﻳﻲ  داروﺳﺎزي ﻋﻤﻮﻣﻲﭘﺎﻳﺎن ﻧﺎﻣﻪ دﻛﺘﺮاي   ﻋﻨﻮان: ﻴﺐدر ﺗﺮﻛ ﻴﻞﻓﻨ ﻲﻣﺘﻮﻛﺴ يﺗﺮ- 3،4،5ﻓﺎرﻣﺎﻛﻮﻓﻮر  ﻴﺖاﻫﻤ ﻲﺑﺮرﺳ ‐5،4،3([‐5‐}onima]enedilyhtem)lynehporolhcid‐3،2([{‐4 enoiht‐3‐elozairt‐4،2،1‐H2‐]lyhtem )lynehpyxohtemirt  ﻴﻦﺗﻮﺑﻮﻟ ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﻲو ﻣﻬﺎر ﻛﻨﻨﺪﮔ ﻴﺘﻮﺗﻮﻛﺴﻴﻚاﺛﺮات ﺳ ﺑﺮ  ﺗﻮﺳﻂ: اﺳﻔﻨﺪﻳﺎريﻣﺮﻳﻢ   راﻫﻨﻤﺎ: اﺳﺎﺗﻴﺪ  دﻛﺘﺮ ﻋﺎﻟﻴﻪ ﻋﺎﻣﺮي  ﺣﻤﻴﺪ ﻓﺮوﺗﻦ ﻓﺮ دﻛﺘﺮ      ﺷﻤﺎره ﭘﺎﻳﺎن ﻧﺎﻣﻪ:                         7931ﻣﺎه آذر 1301    Kerman University of Medical Sciences Faculty of Pharmacy Pharm. D Thesis  Title: Evaluation of the importance of ٣،٤،٥-trimethoxyphenyl pharmacophore of ‐{[(٢،٣‐dichlorophenyl)methylidene]amino} ‐٥‐[(٣،٤،٥‐trimethoxyphenyl)methyl]‐٢H‐١،٢،٤‐triazole‐٣‐thione for cytotoxicity effects and inhibition of tubulin polymerization  By: Maryam Esfandiari   Supervisors: Dr. Alieh Ameri Dr. Hamid Forootanfar        December ٢٠١٨                                                        :ﻲﺧﻼﺻﻪ ﻓﺎرﺳﻫﺎي ﻃﺮاﺣﻲ و ﺳﻨﺘﺰ ﺗﺮﻛﻴﺒﺎت ﺑﺎ ﻣﻜﺎﻧﻴﺴﻤﻬﺎي ﺟﺪﻳﺪ ﻣﻬﺎر رﺷﺪ ﺳﻠﻮﻟﻬﺎي ﺳﺮﻃﺎﻧﻲ ﺑﻪ ﻳﻜﻲ از زﻣﻴﻨﻪ ﻫﺪف:ﺣﻀﻮر ﺗﺤﻘﻴﻘﺎﺗﻲ ﻣﻬﻢ ﺗﺒﺪﻳﻞ ﺷﺪه اﺳﺖ. در ﺗﺤﻘﻴﻖ ﺣﺎﺿﺮ ﻛﻪ در اداﻣﻪ ﭘﺮوژه ﻫﺎي ﻗﺒﻠﻲ ﺻﻮرت ﮔﺮﻓﺘﻪ، اﻫﻤﻴﺖ ( در اﻳﺠﺎد اﺛﺮ ﺳﻤﻴﺖ ﺳﻠﻮﻟﻲ و ﻗﺪرت ﻣﻬﺎر ﻛﻨﻨﺪﮔﻲ ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن PMTﺗﺮي ﻣﺘﻮﻛﺴﻲ ﻓﻨﻴﻞ )-3،4،5ﻓﺎرﻣﺎﻛﻮﻓﻮر  ﺗﻮﺑﻮﻟﻴﻦ ﻳﻚ ﺳﺮي از ﺷﻴﻒ ﺑﺎزﻫﺎي ﻫﻴﺒﺮﻳﺪي ﻣﻮرد ارزﻳﺎﺑﻲ ﻗﺮار ﮔﺮﻓﺘﻪ اﺳﺖ.اراﺋﻪ ﺷﺪه در ﺷﻜﻞ زﻳﺮ ﺳﻨﺘﺰ ﺷﺪﻧﺪ و ﺧﺼﻮﺻﻴﺎت آﻧﻬﺎ از  ﻃﺮحﺷﻴﻒ ﺑﺎزﻫﺎي ﻃﺮاﺣﻲ ﺷﺪه ﺑﺎ  ﻫﺎ: ﻣﻮاد و روشآﻧﻬﺎ ﺗﻬﻴﻪ ﮔﺮدﻳﺪ. ﻣﻴﺰان ﺳﻤﻴﺖ ﺳﻠﻮﻟﻲ ﺗﺮﻛﻴﺒﺎت ﺳﻨﺘﺰ  RMNC31و  RMNH1، RITFﺟﻤﻠﻪ ﻧﻘﻄﻪ ذوب، ﻃﻴﻔﻬﺎي ﻛﻪ ﺑﻪ ﺗﺮﺗﻴﺐ ﻣﺘﻌﻠﻖ ﺑﻪ ﺳﺮﻃﺎﻧﻬﺎي ﻓﻴﺒﺮوﺳﺎرﻛﻮﻣﺎ،  945Aو  7FCM، 2GpeH، 0801THرده ﺳﻠﻮﻟﻲ  4ﺷﺪه ﻋﻠﻴﻪ ( در ﺣﻀﻮر %ytilibaiVﺑﺮرﺳﻲ و درﺻﺪ زﻧﺪه ﻣﺎﻧﻲ )  yassA TTMﺪ ﺑﻪ ﻛﻤﻚ روشﻛﺒﺪ، ﭘﺴﺘﺎن و رﻳﻪ ﻣﻲ ﺑﺎﺷﻨﻣﺮﺑﻮﻃﻪ ﻣﺤﺎﺳﺒﻪ ﮔﺮدﻳﺪ. در ﻣﺮﺣﻠﻪ ﺑﻌﺪ ﺗﺮﻛﻴﺒﺎت واﺟﺪ  05CIﻏﻠﻈﺘﻬﺎي ﻣﺨﺘﻠﻒ اﻳﻦ ﺗﺮﻛﻴﺒﺎت و ﻫﻤﭽﻨﻴﻦ ﻣﻘﺎدﻳﺮ ﻣﻨﺎﺳﺐ اﻧﺘﺨﺎب و ﺗﻮاﻧﺎﻳﻲ آﻧﻬﺎ در ﻣﻬﺎر ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﺗﻮﺑﻮﻟﻴﻦ ﺑﻪ ﻛﻤﻚ ﻛﻴﺖ ﻣﺘﻌﻠﻖ ﺑﻪ ﺷﺮﻛﺖ  05CIﻣﺮﺑﻮﻃﻪ در ﺣﻀﻮر ﻏﻠﻈﺘﻬﺎي ﻣﺨﺘﻠﻒ ﺗﺮﻛﻴﺒﺎت  05CI. درﺻﺪ ﻣﻬﺎر ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن و ﺷﺪﺑﺮرﺳﻲ  noteleksotyCﻣﻨﺘﺨﺐ ﻣﺤﺎﺳﺒﻪ ﮔﺮدﻳﺪ. در ﻣﺮﺣﻠﻪ ﺑﻌﺪ و ﺑﻪ ﻣﻨﻈﻮر ﺑﺮرﺳﻲ ﻣﻜﺎﻧﻴﺴﻢ اﺣﺘﻤﺎﻟﻲ اﺗﺼﺎل و ﻣﺤﺎﺳﺒﻪ اﻧﺮژﻳﻬﺎي ﭘﻴﻮﻧﺪ   .ﺷﺪاﺳﺘﻔﺎده  0AS1 و ﺳﺎﺧﺘﺎر ﻛﺮﻳﺴﺘﺎﻟﻲ ﺗﻮﺑﻮﻟﻴﻦ ﺑﺎ ﻛﺪ kcoDotuA ﺗﺮﻛﻴﺒﺎت ﻣﻨﺘﺨﺐ ﺑﺎ ﺗﻮﺑﻮﻟﻴﻦ از ﻧﺮم اﻓﺰار    esab ffihcSﺳﻨﺘﺰ ﻣﺸﺘﻘﺎت  -1ﻃﺮح  ﺑﺮرﺳﻲ ﻃﻴﻒ ﺳﻨﺠﻲ ﺻﻮرت ﮔﺮﻓﺘﻪ ﻧﺸﺎن داد ﻛﻪ ﺗﺮﻛﻴﺒﺎت ﻣﻮرد ﻧﻈﺮ ﺳﻨﺘﺰ ﺷﺪه اﻧﺪ. ﻧﺘﺎﻳﺞ ارزﻳﺎﺑﻲ ﺳﻤﻴﺖ : ﻫﺎ ﻳﺎﻓﺘﻪﻋﻠﻴﻪ رده ﺳﻠﻮﻟﻲ  ، 05CI = 57/6±02/5 Mμ) a-30-Mﻧﺸﺎن داد ﻛﻪ ﻓﻘﻂ ﺗﺮﻛﻴﺐ  30-Mﺳﻠﻮﻟﻲ ﺗﺮﻛﻴﺒﺎت دﺳﺘﻪ ﻪ اي ﺑﺮوز ـ( از اﻳﻦ ﮔﺮوه ﻧﺘﻴﺠﻪ ﻣﻨﺎﺳﺐ ﻧﺸﺎن داد و ﺳﺎﻳﺮ ﺗﺮﻛﻴﺒﺎت اﻳﻦ ﮔﺮوه ﺳﻤﻴﺖ ﺳﻠﻮﻟﻲ ﻗﺎﺑﻞ ﻣﻼﺣﻈ0801THﻲ ـﻠﻮﻟـﺮ روي رده ﺳـﺑ d-40-Mو  b-40-Mو  a-40-Mﺳﻪ ﺗﺮﻛﻴﺐ  40-Mﺒﺎت دﺳﺘﻪ ـﻧﺪادﻧﺪ. از ﺑﻴﻦ ﺗﺮﻛﻴﻧﺸﺎن دادﻧﺪ. اﻳﻦ ﺳﻪ  441/67±4/01 Mμو  23/2±3/2 Mμ،  51/7±0/80 Mμ ﺎدلـﻣﻌ 05CIﻴﺐ ـﺑﻪ ﺗﺮﺗ 2GpeHﺗﺮﻛﻴﺐ ﺑﻪ ﻫﻤﺮاه ﻛﻠﺸﻲ ﺳﻴﻦ ﺑﻪ ﻋﻨﻮان ﻛﻨﺘﺮل ﻣﺜﺒﺖ در ﺗﺴﺖ ﻣﻬﺎرﻛﻨﻨﺪﮔﻲ ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﺗﻮﺑﻮﻟﻴﻦ ﻣﻮرد ﺑﺮرﺳﻲ  Mμو  63/27±1/37 Mμ، 11/81±0/32 Mμ، 0/71±0/10 Mμ آﻧﻬﺎ ﺑﻪ ﺗﺮﺗﻴﺐ 05CIﻗﺮار ﮔﺮﻓﺘﻨﺪ ﻛﻪ ﻣﻴﺰان   و ﻛﻠﺸﻲ ﺳﻴﻦ ﻣﺤﺎﺳﺒﻪ ﮔﺮدﻳﺪ. d-40-Mو  b-40-Mو  a-40-Mﺑﺮاي ﺗﺮﻛﻴﺒﺎت  0/91±0/10: ﺑﺮرﺳﻲ ﻫﺎي ﺻﻮرت ﮔﺮﻓﺘﻪ در اﻳﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن داد ﻛﻪ ﺑﻴﻦ ﻣﻴﺰان ﺳﻤﻴﺖ ﺳﻠﻮﻟﻲ و اﺛﺮ ﻣﻬﺎر ﻛﻨﻨﺪﮔﻲ ﮔﻴﺮي ﻧﺘﻴﺠﻪو ﻫﻤﭽﻨﻴﻦ ﻧﺘﺎﻳﺞ ﺣﺎﺻﻞ از داﻛﻴﻨﮓ ﺗﺮﻛﻴﺒﺎت از ﻟﺤﺎظ ﻣﻴﺰان اﻧﺮژي و ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﺗﻮﺑﻮﻟﻴﻦ ﺗﺮﻛﻴﺒﺎت ﺳﻨﺘﺰ ﺷﺪه راﺑﻄﻪ ﻣﻨﺎﺳﺒﻲ وﺟﻮد داﺷﺖ و ﺣﻀﻮر در ﺟﺎﻳﮕﺎه اﺗﺼﺎل ﺗﻮﺑﻮﻟﻴﻦ  142ﺗﻮاﻧﺎﻳﻲ اﺗﺼﺎل ﺑﻪ اﺳﻴﺪ آﻣﻴﻨﻪ ﺳﻴﺴﺘﺌﻴﻦ ﻣﻬﻢ و ﺿﺮوري ﻣﻲ  ﻴﻦﺗﻮﺑﻮﻟ ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﻲﻣﻬﺎر ﻛﻨﻨﺪﮔﺟﻬﺖ ﺑﺮوز اﺛﺮات ﺳﻤﻴﺖ ﺳﻠﻮﻟﻲ و  PMTﻓﺎرﻣﺎﻛﻮﻓﻮر   ﺑﺎﺷﺪ. ﻴﻦﺗﻮﺑﻮﻟ ﭘﻠﻴﻤﺮﻳﺰاﺳﻴﻮن ﻲﻣﻬﺎر ﻛﻨﻨﺪﮔ ﻲ،ﺳﻠﻮﻟ ﻴﺖﺳﻤ ﻴﺲ،ﺑ ﻴﻒ، ﺷ ﻴﻞﻓﻨ ﻲﻣﺘﻮﻛﺴ يﺗﺮ-3،4،5 ﻛﻠﻴﺪي: ﻛﻠﻤﺎت    :tcartsbA gnitibihni fo smsinahcem levon eht htiw sdnuopmoc fo sisehtnys dna ngised ehT :esopruP eht saw hcihw ,yduts tneserp eht nI .hcraeser fo dleif tnatropmi na emoceb sah htworg llec recnac-5،4،3 erohpocamrahp fo ecneserp eht fo ecnatropmi eht ,seiduts suoiverp eht fo gniwollof nilubut fo ytivitca yrotibihni dna ytivitca cixototyc eht no )PMT( lynehpyxohtemirt .serutcurts dirbyh fo seires a ni detaulave saw noitaziremylop emehcs eht ot gnidrocca dezisehtnys erew sesab ffihcS dengised ehT :sdohtem dna slairetaM RMNC31 dna ,RMNH1 ,RITF ,tniop gnitlem gnidulcni scitsiretcarahc rieht dna woleb detneserpspectra were determined. The cytotoxicity of the synthesized compounds against the four cell lines of HT1080, HepG2, MCF7, and A549 belonging to fibrosarcoma, liver, breast, and lung cancers, were investigated using the MTT assay method, and the percentage of viability in the presence of different concentrations of these compounds as well as the corresponding IC50 values were calculated. In the next step, the compounds with appropriate IC50 were selected and their ability to inhibit tubulin polymerization by means of the kit belonging to the Cytoskeleton Company was investigated. The percentage of inhibition of polymerization and corresponding IC50 in the presence of different concentrations of the selected compounds were calculated. In the next step, the AutoDock software and crystalline structure of tubulin with the PDB code 1SA0 was used to examine the mechanism of probable binding and calculation of binding energy of the selected compounds to tubulin.        ج    Scheme 1- Synthesis of Schiff base derivatives  Results: The obtained spectra of attained compounds showed that the desired compounds were synthesized. The results of cytotoxicity effect of M-03 compounds revealed that only M-03-a (IC50 = 75,6 ± 20,5 μM against the HT1080 cell line) exhibited suitable inhibitory effect and the other compounds of this group did not show appropriate cytotoxic effect. Among compounds of the M-04 group, the three compounds of M-04-a, M-04-b, and M-04-d inhibited the HepG2 cell line by IC50 of 15,70,8 μM, 32,23,2 μM, and 144,764,1 μM, respectively. Thereafter, the tubulin polymerization inhibitory activity of these three compounds in combination with colchicine, as positive control of tubulin inhibitor was investigated. The related IC50 values were found to be 0,170,01 μM, 11,180.23 μM, 36,721,73 μM, and 0,19 ± 0,01 μM for M-04-a, M-04-b, M-04-d and colchicine, respectively. Conclusion: To sum up, the obtained results of the present study showed suitable correlation between cytotoxicity effect and tubulin polymerization inhibitory activity as well as the results of docking studies (in terms of binding energy and the ability to interact with Cys241 of tubulin binding site of tubulin).  Keywords: 3،4،5-trimethoxyphenyl, Schiff base, cytotoxicity, tubulin polymerization inhibitory activity        

=Evaluation of the importance of 3,4,5-trimethoxyphenyl pharmacophore of ‐{[(2,3‐dichlorophenyl)methylidene]amino} ‐5‐[(3,4,5‐trimethoxyphenyl)methyl]‐2H‐1,2,4‐triazole‐3‐thione for cytotoxicity effects and inhibition of tubulin polymerization

http://eprints.kmu.ac.ir/29745/1/6935.pdf

=Evaluation of the importance of 3,4,5-trimethoxyphenyl pharmacophore of ‐{[(2,3‐dichlorophenyl)methylidene]amino} ‐5‐[(3,4,5‐trimethoxyphenyl)methyl]‐2H‐1,2,4‐triazole‐3‐thione for cytotoxicity effects and inhibition of tubulin polymerization

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