Potensi senyawa(1)-N-(3,4-dimetoksibenzoil)1,10-fenantrolin-bromida dan senyawa (1)-N-(1-metoksibenzoil)-1,10-fenantrolin-bromida sebagai antijamur: kajian aktivitas dan toksisitas in vitro dan kemungkinan mekanisme aksinya

MAULINA DIAH SUKARLAN, DRA., APT., M.KES, Prof. Dr. Mustofa, M. Kes, Apt

2016 | Disertasi | S3 Ilmu Kedokteran

Latar Belakang. Candida sp adalah flora normal dijumpai dalam organ manusia. Namun Candida sp dapat bersifat patogen karena kemampuannya menginfeksi sel inang dan mengakibatkan penyakit apabila kondisi sistem imun inang lemah. Senyawa turunan 1.10 fenantrolin merupakan kelator logam yang memiliki efek antijamur. Pada studi pendahuluan senyawa (1)-N-2 metoksibenzil-1.10 fenantrolin bromide dan senyawa (1)-N-(1-metoksibenzoil)-1,10-fenantrolin bromida menghambat pertumbuhan jamur C. albicans. Tujuan. Mengkaji aktivitas antijamur in vitro senyawa 1, senyawa 2 dan flukonazol terhadap C. albicans . Mengkaji toksisitas in vitro nya pada kultur sel normal selain itu juga mengkaji pengaruhnya terhadap penghambatan pembentukan dan pengurangan pembentukan biofilm C. albicans dan mengkaji integritas membran sel C. albicans. Hasil Penelitian. Penelitian KHM senyawa 1 dan senyawa 2 terhadap C. albicans ATCC 10231, isolat klinik Candida 2, Candida 20 antara 0,78-3.13 mikrogram/mL. KHM flukonazol antara 0,25-0,50 mikrogram/mL. Penelitian KBM senyawa 1, 2 dan flukonazol terhadap C. albicans ATCC 10231 berturut-turut adalah 100 mikrogram/mL, 100 mikrogram/mL dan >64 mikrogram/mL. Senyawa 1, 2 dan flukonazol terhadap Candida 2 adalah >100 mikrogram/mL >100 mikrogram/mL dan >64 mikrogram/mL. Senyawa 1, 2 dan flukonazol terhadap Candida 20 adalah >64 mikrogram/mL, >64 mikrogram/mL dan >64 mikrogram/mL. Penelitian Inhibitory Concentration 50 (IC 50) senyawa 1,2 pada Candida 2 adalah 56,17 plus/minus 21,59 mikrogram/mL; 84,93 plus/minus 21,09 mikrogram/mL dan flukonazol > 1000 mikrogram/mL. IC50 senyawa 1 dan 2 pada Candida 20 adalah 56,17 plus/minus 21,59 mikrogram/mL ; 84,93 plus/minus 21,09 mikrogram/mL dan flukonazol > 1000 mikrogram/mL. IC 50 senyawa 1, 2 pada C. albicans adalah 84,93 plus/minus 21,09 mikrogram/mL 56,17 plus/minus 21,59 mikrogram/mL dan flukonazol > 1000 mikrogram/mL. Indek sitotoksik ( IS) senyawa 1, 2 dan flukonazol pada Candida 2 berturut turut 17,9 ; 27,1 dan > 4000. IS senyawa 1, 2 dan flukonazol pada Candida 20 dan flukonazol berturut turut 54,44 ; 36,0 dan > 2000. IS senyawa 1, 2 dan flukonazol pada C. albicans berturut turut 54,44 ; 36,0 dan > 2000. Penelitian Kadar Hambat Minimal (KHBM) Nilai KHBM50 C. albicans, Candida 2 dan Candida 20 adalah 0,41;0,72 dan 0,48 mikrogram/mL. Nilai KHBM80 senyawa 1 pada C. albicans, Candida 2 dan Candida 20 adalah 0,21 ; < 0,04 dan < 0,04 mikrogram/mL. Nilai KHBM50 senyawa 2 pada C. albicans, Candida 2 dan Candida 20 adalah 1,53 ; 1,21 dan 0,82 mikrogram/mL. Nilai KHBM80 senyawa 2 pada C. albicans, Candida 2 dan Candida 20 adalah < 0,80 0,29; 0,39 mikrogram/mL. Nilai KHBM50 flukonazol pada C. albicans, Candida 2 dan Candida 20 antara 0,53 ; 1,46 dan 0,83 mikrogram/mL. Nilai KHBM80 flukonazol pada C. albicans, Candida 2 dan Candida 20 adalah 0,18 ; 0,40 dan 0,30 mikrogram/mL. Penelitian Kadar Reduksi Biofilm Minimal (KRBM). Nilai KRBM50 senyawa 1 pada C. albicans, Candida 2 dan Candida 20 berkisar antara 1,59 ; 3,67 dan 2,48 mikrogram/mL. KRBM80 senyawa 1 pada C. albicans, Candida 2 dan Candida 20 adalah 0,71 ; 1,79 dan 1,93 mikrogram/mL. KRBM50 senyawa 2 pada C. albicans, Candida 2 dan Candida 20 adalah 1,08 - 3,89 dan 1,24 mikrogram/mL. KRBM80 senyawa 2 pada C. albicans, Candida 2 dan Candida 20 adalah 0,47 ; 2,31 dan 0,69 mikrogram/mL. KRBM50 flukonazol pada C. albicans, Candida 2 Candida 20 adalah 0,60 ; 2,58 dan 1,09 mikrogram/mL. KHBM80 flukonazol pada ketiga isolat C. albicans antara 0,25 ; 0,74 dan 1,09 mikrogram/mL. Penelitian integritas membran sel Candida 2, peningkatan dosis senyawa 1 dan 2 yaitu 2 x KHM, 4 x KHM dan 8 x KHM sebagai berikut: senyawa 1 dengan nilai berturut-turut 46,50 plus/minus 1,57 mikrogram/mL ; 50,00 plus/minus 3,01 mikrogram/mL ; 69,62 plus/minus 1,12 mikrogram/mL. Sementara senyawa 2 memiliki nilai berturut-turut adalah 58,71 plus/minus 0,71 mikrogram/mL; 67,77 plus/minus 0,35 mikrogram/mL; 63,34 plus/minus 9,43 mikrogram/mL. Pada flukonazol dengan konsentrasi 8 x Kadar Hambat Minimal (KHM) adalah 51,47 plus/minus 1,29 mikrogram/mL. Pada Candida 20, peningkatan dosis senyawa 1 dan 2 (2 x Kadar Hambat Minimal (KHM), 4 x Kadar Hambat Minimal (KHM) dan 8 x Kadar Hambat Minimal (KHM) sebagai berikut senyawa 1 dengan nilai berturut-turut 9,77 plus/minus 0,27 mikrogram/mL; 19,91 plus/minus 0,21 mikrogram/mL; dan 21,60 plus/minus 0,65 mikrogram/mL. Sementara senyawa 2 memiliki nilai berturut-turut 20,95 plus/minus 0,45 mikrogram/mL; 32,39 plus/minus 0,66 mikrogram/mL dan 27,55 plus/minus 0,25 mikrogram/mL. Pada flukonazol dengan konsentrasi 8x Kadar Hambat Minimal (KHM) adalah 27,36 plus/minus 0,78 mikrogram/mL. Pada Candida albicans ATCC 10231 peningkatan dosis senyawa 1 dan 2 (2 x Kadar Hambat Minimal (KHM), 4 x Kadar Hambat Minimal (KHM) dan 8 x Kadar Hambat Minimal (KHM): senyawa 1 dengan nilai berturut-turut 13,08 plus/minus 0,62 mikrogram/mL; 42,03 plus/minus 0,91mikrogram/mL dan 31,16 plus/minus 0,36 mikrogram/mL. Sementara senyawa 2 memiliki nilai berturut-turut 26,91 plus/minus 0,44 mikrogram/mL; 20,63 plus/minus 0,15 mikrogram/mL dan 65,94 plus/minus 1,20 mikrogram/mL. Pada flukonazol dengan konsentrasi 8 x Kadar Hambat Minimal (KHM) adalah 28,02 plus/minus 0,30 mikrogram/mL. Kelipatan dosis senyawa 1 dan 2 tidak selalu berefek meningkatkan kerusakan membran sel C. albicans ATCC 10231, isolat klinik Candida 2 dan Candida 20. Senyawa 1, senyawa 2 dan flukonazol menghambat dan membunuh C. albicans ATCC 10231 isolat klinik Candida 2 dan Candida 20. Senyawa 1, senyawa 2 dan flukonazol menghambat pertumbuhan sel Vero sampai 50 % terhadap C. albicans ATCC 10231, isolat klinik Candida 2 dan Candida 20. Senyawa 1, senyawa 2 dan flukonazol aman terhadap C. albicans ATCC 10231, isolat klinik Candida 2 dan Candida 20, terlihat pada nilai indek sitotoksiknya. Senyawa 1, senyawa 2 dan flukonazol menghambat dan mengurangi pertumbuhan C. albicans ATCC 10231, isolat klinik Candida 2 dan Candida 20. Kenaikan dosis senyawa 1 dan senyawa 2 tidak selalu meningkatkan kerusakan dinding sel C. albicans ATCC 10231, isolat klinik Candida 2 dan Candida 20.

Background. The 1.10 phenantroline derivative compound is a metal-based drugs possessing effects as an anti plasmodium, anti-cancer and anti-fungal. In a preliminary study of the compound (1)-N-2-methoxy-benzyl 1.10 phenantroline bromide proven to inhibit the growth of fungi Candida spp. Aim. Reviewing invitro activity of the anti-fungal, the in vitro toxicity in normal cell culture, anti-fungal activity through its influence on the formation of biofilm and membrane integrity of the (1)-N-(3,4-dimethoxy benzyl -1.10fenantrolin bromide [1st compound], (1)-N-(1-methoxy-3methoxy benzyl -1.10 fenantrolin bromide [2nd compound] and fluconazole against Candida albicans ATCC 10231, clinical isolates of Candida 2 and 20. Results. MIC values of the 1st and 2nd compounds towards Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20 were between 0,78-3.13 microgram/mL, while the MIC values of fluconazole were between 0,25-0,50 microgram/mL. The research on MBC values of 1st and 2nd compounds and fluconazole towards Candida albicans ATCC 10231 were 100 microgram/mL, 100 microgram/mL and >64 microgram/mL, respectively. As the MBC values of 1st and 2nd compounds and fluconazole towards Candida 2 were >100 microgram/mL >100 microgram/mL and >64 microgram/mL, consecutively; while the MBC values of 1st and 2nd compounds and fluconazole towards Candida 20 were >64 microgram/mL, >64 microgram/mL and >64 microgram/mL, respectively. The research on Inhibitory Concentration50 (IC50) values of 1st and 2nd compounds for Candida 2 were 56,17 plus/minus 21,59 microgram/mL; 84,93 plus/minus 21,09 microgram/mL as fluconazole was >1000 microgram/mL. IC50 values on 1st and 2nd compounds for Candida 20 were 56,17 plus/minus 21,59 microgram/mL; 84,93 plus/minus 21,09 microgram/mL as fluconazole were >1000 microgram/mL, while the IC50 values for Candida albicans were 84,93 plus/minus 21,09 microgram/mL; 56,17 plus/minus 21,59 microgram/mL and fluconazole was >1000 microgram/mL. Index Cytotoxic (IC) of the 1st and 2nd compounds, and fluconazole towards Candida 2 consecutively were 17,9; 27,1 and > 4000. The IC values towards Candida 20, respectively were 54,44; 36,0 and > 2000, while IC values towards Candida albicans were 54,44; 36,0 and > 2000. The results of MBIC50 values 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,41;0,72 and 0,48 microgram/mL. The MBIC80 values of 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,21; < 0,04 and < 0,04 microgram/mL. The MBIC50 of 2nd compound for Candida albicans, Candida 2 and Candida 20 were 1,53; 1,21 and 0,82 microgram/mL. The MBIC80 2nd compound for Candida albicans, Candida 2 and Candida 20 were < 0,80; 0,29; and 0,39 microgram/mL. The MBIC50 fluconazole for Candida albicans, Candida 2 and Candida 20 were between 0,53; 1,46 and 0,83 microgram/mL. MBIC80 values for fluconazole for Candida albicans, Candida 2 and Candida 20 were 0,18; 0,40 and 0,30 microgram/mL. The MBRC50 values of 1st compound for Candida albicans, Candida 2 and Candida 20 ranged between 1,59; 3,67 and 2,48 microgram/mL. MBRC80 values of 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,71; 1,79 and 1,93 microgram/mL. fluconazole was >1000 microgram/mL. IC50 values on 1st and 2nd compounds for Candida 20 were 56,17 plus/minus 21,59 microgram/mL; 84,93 plus/minus 21,09 microgram/mL as fluconazole were >1000 microgram/mL, while the IC50 values for Candida albicans were 84,93 plus/minus 21,09 microgram/mL; 56,17 plus/minus 21,59 microgram/mL and fluconazole was >1000 microgram/mL. Index Cytotoxic (IC) of the 1st and 2nd compounds, and fluconazole towards Candida 2 consecutively were 17,9; 27,1 and > 4000. The IC values towards Candida 20, respectively were 54,44; 36,0 and > 2000, while IC values towards Candida albicans were 54,44; 36,0 and > 2000. The results of MBIC50 values 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,41;0,72 and 0,48 microgram/mL. The MBIC80 values of 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,21; < 0,04 and < 0,04 microgram/mL. The MBIC50 of 2nd compound for Candida albicans, Candida 2 and Candida 20 were 1,53; 1,21 and 0,82 microgram/mL. The MBIC80 2nd compound for Candida albicans, Candida 2 and Candida 20 were < 0,80; 0,29; and 0,39 microgram/mL. The MBIC50 fluconazole for Candida albicans, Candida 2 and Candida 20 were between 0,53; 1,46 and 0,83 microgram/mL. MBIC80 values for fluconazole for Candida albicans, Candida 2 and Candida 20 were 0,18; 0,40 and 0,30 microgram/mL. The MBRC50 values of 1st compound for Candida albicans, Candida 2 and Candida 20 ranged between 1,59; 3,67 and 2,48 microgram/mL. MBRC80 values of 1st compound for Candida albicans, Candida 2 and Candida 20 were 0,71; 1,79 and 1,93 microgram/mL On Candida albicans ATCC 10231 For fluconazole with 8 x MIC were as follows: 1st compound with consecutive values 13,08 plus/minus 0,62 microgram/mL; 42,03 plus/minus 0,91 microgram/mL and 31,16 plus/minus 0,36 microgram/mL. As for 2nd compound were 26,91 plus/minus 0,44 microgram/mL; 20,63 plus/minus 0,15 microgram/mL and 65,94 plus/minus 1,20 microgram/mL, respectively. For fluconazole with 8 x MIC were 28,02 plus/minus 0,30 microgram/mL. The multiplication dose of the 1st and the 2nd compounds does not constantly increasing the damaging effect on Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20 membrane cells integrity. The 1st and the 2nd compounds, as well as fluconazole inhibits and killed Candida albicans ATCC 10231, clinical isolates Candida 2 and Candida 20. The 1st and the 2nd compounds, as well as fluconazole inhibits the growth of vero cell until 50 % towards Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20. The 1st and the 2nd compounds, as well as fluconazole were considered safe towards Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20 as seen on the Cytotoxic Index. The increasing doses of the 1st and the 2nd compounds, as well as fluconazole inhibits the growth of vero cell until 50% towards Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20. The multiplication dose of the 1st and the 2nd compounds does not constantly increasing the damaging effect on Candida albicans ATCC 10231, clinical isolates of Candida 2 and Candida 20 membrane cells integrity.

Kata Kunci : senyawa (1)-N-(3,4-dimetoksibenzoil)-1,10-fenantrolin-bromida dan senyawa (1)-N-(1-metoksibenzoil)-1,10-fenantrolin-bromida, flukonazol, KHM, KBM, sitotoksisitas, KHBM, KRBM, integritas membran