ºðºì²ÜÆ äºî²Î²Ü вزÈê²ð²ÜÆ ¶Æî²Î²Ü îºÔºÎ²¶Æð Ó×ÅÍÛÅ ÇÀÏÈÑÊÈ ÅÐÅÂÀÍÑÊÎÃÎ ÃÎÑÓÄÀÐÑÒÂÅÍ ÍÎÃÎ ÓÍÈÂÅÐ ÑÈÒÅÒÀ ´Ý³Ï³Ý ·ÇïáõÃÛáõÝÝ»ñ êè 1, 2007 Åñòåñòâåííûå íàó- Ȼɢ ɨ ɥ ɨ ɝ ɢ ɹ ɍȾɄ 612.014.4.083.36 Ɋ. Ɇ. ȺɊɍɌɘɇəɇ, Ƚ. Ƚ. ɈȽȺɇȿɋəɇ, Ɋ. ȿ. ɆɍɊȺȾəɇ, Ʉ. ɋ. ɆȺɊȽȺɊəɇ ɋɊȺȼɇɂɌ ȿɅɖɇɕɃ ȺɇȺɅɂɁ ȽȿɇɈɌ ɈɄɋɂɑȿɋɄɂɏ ɗɎɎȿɄɌ Ɉȼ ɇɈȼɕɏ ɉɈɊɎɂɊɂɇɈȼ ɆȿɌ ɈȾɈɆ ȾɇɄ-ɄɈɆȿɌ IN VIVO Ɇɟɬɨɞɨɦ ȾɇɄ-ɤɨɦɟɬ ɨɰɟɧɢɜɚɥɚɫɶ ɫɪɚɜɧɢɬɟɥɶɧɚɹ ɝɟɧɨɬɨɤɫɢɱɧɨɫɬɶ ɧɨɜɨɝɨ ɩɨɪɮɢɪɢɧɚ TOEt4PyP ɢ ɟɝɨ Ag-, Zn- ɢ ɋɨ-ɩɪɨɢɡɜɨɞɧɵɯ ɜ ɥɟɣɤɨɰɢɬɚɯ ɤɪɨɜɢ ɦɵɲɟɣ in vivo ɜ ɡɚɜɢɫɢɦɨɫɬɢ ɨɬ ɜɪɟɦɟɧɢ ɜɨɡɞɟɣɫɬɜɢɹ. ɉɨɜɵɲɟɧɢɟ ɭɪɨɜɧɹ ɩɨɜɪɟɠɞɟɧɢɣ ȾɇɄ ɱɟɪɟɡ 24 ɱ ɩɨɫɥɟ ɜɜɟɞɟɧɢɹ ɢɡɭɱɚɟɦɵɯ ɩɨɪɮɢɪɢɧɨɜ ɩɨ ɫɪɚɜɧɟɧɢɸ ɫ ɭɪɨɜɧɟɦ, ɧɚɛɥɸɞɚɟɦɵɦ ɱɟɪɟɡ 6 ɱɚɫɨɜ, ɫɜɢɞɟɬɟɥɶɫɬɜɭɟɬ ɨ ɤɭɦɭɥɹɬɢɜɧɨɦ ɷɮɮɟɤɬɟ ɢɯ ɜɨɡɞɟɣɫɬɜɢɹ, ɱɬɨ ɦɨɠɟɬ ɛɵɬɶ ɢɫɩɨɥɶɡɨɜɚɧɨ ɜ ɯɢɦɢɨɬɟɪɚɩɢɢ ɨɩɭɯɨɥɟɣ. ɂɫɫɥɟɞɨɜɚɧɢɹ ɩɨɪɮɢɪɢɧɨɜ ɢ ɢɯ ɚɧɚɥɨɝɨɜ ɡɚ ɩɨɫɥɟɞɧɟɟ ɞɟɫɹɬɢɥɟɬɢɟ ɫɬɪɟɦɢɬɟɥɶɧɨ ɪɚɡɜɢɜɚɸɬɫɹ. ɉɨɜɵɲɟɧɧɵɣ ɢɧɬɟɪɟɫ ɤ ɭɧɢɤɚɥɶɧɨɣ ɫɬɪɭɤɬɭɪɟ ɩɨɪɮɢɪɢɧɨɜ ɢ ɦɟɬɚɥɥɨɩɨɪɮɢɪɢɧɨɜ ɨɛɭɫɥɨɜɥɟɧ ɪɟɚɥɶɧɵɦɢ ɩɟɪɫɩɟɤɬɢɜɚɦɢ ɢɯ ɢɫɩɨɥɶɡɨɜɚɧɢɹ ɜ ɯɢɦɢɨɬɟɪɚɩɢɢ ɨɩɭɯɨɥɟɣ. ȼ ɱɚɫɬɧɨɫɬɢ ɜɟɫɶɦɚ ɩɟɪɫɩɟɤɬɢɜɧɨɣ ɹɜɥɹɟɬɫɹ ɢɯ ɭɧɢɤɚɥɶɧɚɹ ɫɩɨɫɨɛɧɨɫɬɶ ɫɟɥɟɤɬɢɜɧɨ ɧɚɤɚɩɥɢɜɚɬɶɫɹ ɜ ɨɩɭɯɨɥɟɜɵɯ ɬɤɚɧɹɯ ɢ ɭɞɟɪɠɢɜɚɬɶɫɹ ɬɚɦ ɞɥɢɬɟɥɶɧɨɟ ɜɪɟɦɹ [1]. Ɏɚɪɦɚɤɨɥɨɝɢɱɟɫɤɢɟ ɢ ɛɢɨɥɨɝɢɱɟɫɤɢɟ ɫɜɨɣɫɬɜɚ ɩɨɪɮɢɪɢɧɨɜ ɜɨ ɦɧɨɝɨɦ ɨɩɪɟɞɟɥɹɸɬɫɹ ɢɯ ɫɬɪɭɤɬɭɪɨɣ. ɉɨɷɬɨɦɭ ɜɨ ɦɧɨɝɢɯ ɥɚɛɨɪɚɬɨɪɢɹɯ ɡɚɧɢɦɚɸɬɫɹ ɰɟɥɟɧɚɩɪɚɜɥɟɧɧɵɦ ɫɢɧɬɟɡɨɦ ɧɨɜɵɯ ɩɨɪɮɢɪɢɧɨɜ ɫ ɰɟɥɶɸ ɩɨɥɭɱɟɧɢɹ ɧɚɢɛɨɥɟɟ ɚɤɬɢɜɧɵɯ ɢ ɛɟɡɨɩɚɫɧɵɯ ɫɨɟɞɢɧɟɧɢɣ. ɉɨɞɨɛɧɵɣ ɩɨɢɫɤ ɩɪɟɞɩɨɥɚɝɚɟɬ ɨɛɹɡɚɬɟɥɶɧɭɸ ɨɰɟɧɤɭ ɢɯ ɝɟɧɨɬɨɤɫɢɱɧɨɫɬɢ ɫ ɭɱɟɬɨɦ ɦɟɯɚɧɢɡɦɨɜ ɞɟɣɫɬɜɢɹ ɢɡɭɱɚɟɦɵɯ ɫɨɟɞɢɧɟɧɢɣ ɧɚ ȾɇɄ ɢ ɜɨɡɦɨɠɧɨɫɬɢ ɭɦɟɧɶɲɟɧɢɹ ɩɨɛɨɱɧɨɝɨ ɜɨɡɞɟɣɫɬɜɢɹ ɧɚ ɧɨɪɦɚɥɶɧɵɟ ɤɥɟɬɤɢ ɨɪɝɚɧɢɡɦɚ. ɂɫɫɥɟɞɨɜɚɧɢɹ ɬɨɤɫɢɱɧɨɫɬɢ ɢ ɩɪɨɬɢɜɨɨɩɭɯɨɥɟɜɨɣ ɚɤɬɢɜɧɨɫɬɢ ɧɨɜɵɯ ɩɨɪɮɢɪɢɧɨɜ, ɫɢɧɬɟɡɢɪɨɜɚɧɧɵɯ ɧɚ ɤɚɮɟɞɪɟ ɮɚɪɦɚɤɨɥɨɝɢɱɟɫɤɨɣ ɯɢɦɢɢ ȿɪȽɆɍ, ɩɪɨɜɨɞɹɬɫɹ ɜ ɥɚɛɨɪɚɬɨɪɢɢ ɩɪɟɞɤɥɢɧɢɱɟɫɤɨɣ ɬɨɤɫɢɤɨɥɨɝɢɢ ɂɧɫɬɢɬɭɬɚ ɬɨɧɤɨɣ ɨɪɝɚɧɢɱɟɫɤɨɣ ɯɢɦɢɢ ɇȺɇ ɊȺ ɫɨɝɥɚɫɧɨ ɦɟɬɨɞɢɱɟɫɤɢɦ ɪɟɤɨɦɟɧɞɚɰɢɹɦ [2]. Ɉɞɧɨɜɪɟɦɟɧɧɨ ɢɡɭɱɚɸɬɫɹ ɢɯ ɝɟɧɨɬɨɤɫɢɱɟɫɤɢɟ ɫɜɨɣɫɬɜɚ ɦɟɬɨɞɨɦ ȾɇɄ-ɤɨɦɟɬ. Ɇɟɬɨɞ ɲɢɪɨɤɨ ɢɫɩɨɥɶɡɭɟɬɫɹ ɞɥɹ ɨɰɟɧɤɢ ɩɨɜɪɟɠɞɟɧɢɣ ɢ ɪɟɩɚɪɚɰɢɢ ȾɇɄ ɜ ɥɸɛɵɯ ɩɨɩɭɥɹɰɢɹɯ ɷɭɤɚɪɢɨɬɢɱɟɫɤɢɯ ɤɥɟɬɨɤ in vitro ɢ in vivo [3]. ɐɟɥɶɸ ɧɚɫɬɨɹɳɟɝɨ ɢɫɫɥɟɞɨɜɚɧɢɹ ɹɜɥɹɥɨɫɶ ɢɡɭɱɟɧɢɟ ɝɟɧɨɬɨɤɫɢɱɟɫɤɨɣ ɚɤɬɢɜɧɨɫɬɢ ɦɟɡɨ-ɬɟɬɪɚ (4-N-ɨɤɫɢɷɬɢɥɩɢɪɢɞɢɥ) ɩɨɪɮɢɧɚ ɬɟɬɪɚɯɥɨɪɢɞɚ ɢ ɟɝɨ Ag-, Zn- ɢ ɋɨ-ɩɪɨɢɡɜɨɞɧɵɯ ɜ ɤɥɟɬɤɚɯ ɤɪɨɜɢ ɛɟɥɵɯ ɛɟɫɩɨɪɨɞɧɵɯ ɦɵɲɟɣ 153 oT O ET 4P yP C 4P yP Zn TO Et 4P yP Py P Ag TO Et TO ET 4 Ʉ ɨɧ ɬ ɪɨ ɥ ɶ % ȾɇɄɜ ɯɜɨɫɬ ɟ in vivo ɜ ɡɚɜɢɫɢɦɨɫɬɢ ɨɬ ɜɪɟɦɟɧɢ ɜɨɡɞɟɣɫɬɜɢɹ ɩɪɟɩɚɪɚɬɨɜ ɦɟɬɨɞɨɦ ȾɇɄɤɨɦɟɬ (ɝɟɥɶ-ɷɥɟɤɬɪɨɮɨɪɟɡ ɟɞɢɧɢɱɧɵɯ ɤɥɟɬɨɤ). Ɇɚɬɟ ɪɢɚɥ ɢ ɦɟɬɨɞɢɤɚ. ɗɤɫɩɟɪɢɦɟɧɬɵ ɩɪɨɜɨɞɢɥɢ ɧɚ ɛɟɥɵɯ ɛɟɫɩɨɪɨɞɧɵɯ ɦɵɲɚɯ ɜɟɫɨɦ 25–30 ɝ . Ⱦɥɹ ɤɚɠɞɨɝɨ ɷɤɫɩɟɪɢɦɟɧɬɚɥɶɧɨɝɨ ɜɚɪɢɚɧɬɚ ɛɵɥɨ ɢɫɩɨɥɶɡɨɜɚɧɨ ɩɨ 3 ɠɢɜɨɬɧɵɯ. Ɇɵɲɚɦ ɜɜɨɞɢɥɢ ɩɪɟɩɚɪɚɬɵ T OEt4PyP, ZnT OEt4PyP ɢ CoTOEt4PyP ɜ ɞɨɡɟ 25 ɦɝ /ɤɝ ɢ AgT OET4PyP ɜ ɞɨɡɟ 15 ɦɝ /ɤɝ , ɱɬɨ ɫɨɨɬɜɟɬɫɬɜɭɟɬ ½ LD50. Ɂɚɛɨɪ ɨɛɪɚɡɰɨɜ ɰɟɥɶɧɨɣ ɤɪɨɜɢ ɩɪɨɜɨɞɢɥɢ ɱɟɪɟɡ 6 ɢ 24 ɱɚɫɚ ɩɨɫɥɟ ɨɞɧɨɤɪɚɬɧɨɝɨ ɜɜɟɞɟɧɢɹ ɩɪɟɩɚɪɚɬɨɜ. Ⱦɥɹ ɨɰɟɧɤɢ ɢɧɞɭɤɰɢɢ ɤɨɦɟɬ ɩɪɢɦɟɧɹɥɢ ɦɟɬɨɞɢɤɭ ɉ. ɋɢɧɯɚ ɫ ɫɨɚɜɬɨɪɚɦɢ ɫ ɧɟɛɨɥɶɲɢɦɢ ɦɨɞɢɮɢɤɚɰɢɹɦɢ [4]. Ⱦɥɹ ɩɨɥɭɱɟɧɢɹ ɨɩɬɢɦɚɥɶɧɨɣ ɩɥɨɬɧɨɫɬɢ ɤɥɟɬɨɤ ɧɚ ɩɪɟɞɦɟɬɧɨɟ ɫɬɟɤɥɨ ɧɚɧɨɫɢɥɢ ɫɦɟɫɶ 15 ɦɤɥ ɰɟɥɶɧɨɣ ɤɪɨɜɢ ɦɵɲɟɣ ɢ 85 ɦɤɥ ɪɚɫɬɜɨɪɚ ɚɝɚɪɨɡɵ. ȼɫɟ ɨɫɬɚɥɶɧɵɟ ɷɬɚɩɵ ɨɫɭɳɟɫɬɜɥɹɥɢ ɩɨ ɫɬɚɧɞɚɪɬɧɨɣ ɦɟɬɨɞɢɤɟ [5]. ɂɡɨɛɪɚɠɟɧɢɹ ɤɨɦɟɬ ɚɧɚɥɢɡɢɪɨɜɚɥɢ ɫ ɩɪɢɦɟɧɟɧɢɟɦ ɩɪɨɝɪɚɦɦɵ ɚɜɬɨɦɚɬɢɱɟɫɤɨɝɨ ɚɧɚɥɢɡɚ Komet 4 (Kinetic Imaging, Liverpool). ȼ ɤɚɠɞɨɦ ɜɚɪɢɚɧɬɟ ɚɧɚɥɢɡɢɪɨɜɚɥɢ ɩɨ 150 ɤɥɟɬɨɤ. ɍɪɨɜɟɧɶ ɩɨɜɪɟɠɞɟɧɢɣ ȾɇɄ ɨɰɟɧɢɜɚɥɫɹ ɧɚ ɨɫɧɨɜɟ ɢɡɦɟɪɟɧɢɣ ɩɪɨɰɟɧɬɚ ȾɇɄ ɜ ɯɜɨɫɬɟ. ɋɬɚɬɢɫɬɢɱɟɫɤɢɣ ɚɧɚɥɢɡ ɩɨɥɭɱɟɧɧɵɯ ɪɟɡɭɥɶɬɚɬɨɜ ɩɪɨɜɨɞɢɥɢ ɫ ɩɨɦɨɳɶɸ ɩɪɨɝɪɚɦɦ Microsoft EXCEL ɢ ST AT GRAPHICS Plus 5.1 ɫ ɩɪɢɦɟɧɟɧɢɟɦ ɧɟɩɚɪɚɦɟɬɪɢɱɟɫɤɨɝɨ ɬɟɫɬɚ Mann-Whitney (U-test). Ɋɟ ɡɭɥɶɬɚɬɵ ɢ ɨɛɫɭɠɞɟ ɧɢɟ . ɑɟɪɟɡ 6 ɱɚɫɨɜ ɩɨɫɥɟ ɜɜɟɞɟɧɢɹ ɩɪɟɩɚɪɚɬɨɜ ɧɚɛɥɸɞɚɥɨɫɶ ɞɨɫɬɨɜɟɪɧɨɟ (p<0,05) ɩɨɜɵɲɟɧɢɟ % ȾɇɄ ɜ ɯɜɨɫɬɟ ɤɨɦɟɬ ɞɥɹ T OEt4PyP (10,41%), AgT OEt4PyP (9,32%) ɢ ZnTOEt4PyP (8,20%) ɩɨ ɫɪɚɜɧɟɧɢɸ ɫ ɤɨɧɬɪɨɥɟɦ (5,86 %). ɑɟɪɟɡ 24 ɱɚɫɚ ɩɨɫɥɟ ɜɜɟɞɟɧɢɹ ɬɚɤɠɟ ɛɵɥɨ ɨɬɦɟɱɟɧɨ ɞɨɫɬɨɜɟɪɧɨɟ (p<0,05) ɩɨɜɵ14 ɲɟɧɢɟ % ȾɇɄ ɜ ɯɜɨɫɬɟ ɞɥɹ 12 2 ɜɫɟɯ ɢɡɭɱɟɧɧɵɯ ɩɨɪɮɢɪɢɧɨɜ: 10 T OEt4PyP (12,18%), ZnT OEt4PyP 8 (11,94%), CoTOEt4PyP (10,58%) 1 6 ɢ AgT OEt4PyP (10,47%) ɩɨ ɫɪɚɜ4 ɧɟɧɢɸ ɫ ɤɨɧɬɪɨɥɟɦ (5,90 %). 2 ɍɪɨɜɧɢ ɩɨɜɪɟɠɞɟɧɢɣ ȾɇɄ 0 ɱɟɪɟɡ 24 ɱɚɫɚ ɩɨɫɥɟ ɜɜɟɞɟɧɢɹ ɩɨɪɮɢɪɢɧɨɜ ɛɵɥɢ ɞɨɫɬɨɜɟɪɧɨ ɜɵɲɟ, ɱɟɦ ɱɟɪɟɡ 6 ɱɚɫɨɜ (ɫɦ. ɪɢɫɭɧɨɤ). ɇɄ ɜ ɯɜɨɫɬɟ ɤɨɦɟɬ ɜ ɥɟɣɤɨɰɢɬɚɯ ɦɵɲɟɣ ɩɨɫɥɟ Ɍ ɚɤɢɦ ɨɛɪɚɡɨɦ, ɩɨɥɭɱɟɧɜɜɟɞɟɧɢɹ ɩɨɪɮɢɪɢɧɨɜ TOEt4PyP, AgTOEt4PyP, ɧɵɟ ɧɚɦɢ ɪɟɡɭɥɶɬɚɬɵ ɫɜɢɞɟɬɟɥɶZnTOEt4PyP ɢ CoTOEt4PyP: 1 – ɱɟɪɟɡ 6 ɱ ɩɨɫɥɟ ɫɬɜɭɸɬ ɨ ɤɭɦɭɥɹɬɢɜɧɨɦ ɷɮɮɟɤɬɟ ɜɜɟɞɟɧɢɹ; 2 – ɱɟɪɟɡ 24 ɱ. ɢɡɭɱɟɧɧɵɯ ɩɨɪɮɢɪɢɧɨɜ ɩɪɢ ɨɰɟɧɤɟ ɢɯ ɝɟɧɨɬɨɤɫɢɱɧɨɫɬɢ, ɱɬɨ ɫɨɡɞɚɟɬ ɩɪɟɞɩɨɫɵɥɤɢ ɞɥɹ ɢɯ ɜɨɡɦɨɠɧɨɝɨ ɩɪɢɦɟɧɟɧɢɹ ɜ ɯɢɦɢɨɬɟɪɚɩɢɢ ɨɩɭɯɨɥɟɣ. Ɋɚɛɨɬɚ ɜɵɩɨɥɧɟɧɚ ɩɪɢ ɩɨɞɞɟɪɠɤɟ ɝɪɚɧɬɚ Ⱥ-301.2. ɉɨɫɬɭɩɢɥɨ 13.02.2007 Ʉɚɮɟɞɪɚ ɝɟɧɟɬɢɤɢ ɢ ɰɢɬɨɥ ɨɝɢɢ Ʌ ɂ Ɍȿ Ɋ Ⱥ Ɍ ɍɊ Ⱥ 1. Vicente MG .H. – Current Med. Chem., Anti-Cancer Agents, 2001, v.1, p. 175–194. 154 2. ɋɬɟɩɚɧɨɜ Ⱥ.ȼ . Ⱦɨɤɥɢɧɢɱɟɫɤɢɟ ɢɫɫɥɟɞɨɜɚɧɢɹ ɥɟɤɚɪɫɬɜɟɧɧɵɯ ɫɪɟɞɫɬɜ. Ɇɟɬɨɞɢɱɟɫɤɢɟ ɪɟɤɨɦɟɧɞɚɰɢɢ. Ʉɢɟɜ, 2002. 3. T ice R.R., Agurell E., Anderson D., Burlinson B., Hartman A., Kobayashi H., Miyamae Y., Royas E., Ryu J.C., Sasaki Y.F. – Environ. Mol. Mutagen., 2000, v. 35, p. 206–221. 4. Singh N.P. et al. – Exp. Cell Research, 1988, v. 175, p. 184–191. 5. Hovhannisyan G ., Haroutiunian S., G hazaryan R., Aroutiounian R., Margaryan K. – Korean Journal of Environmental Biology, 2005, v. 23, ʋ 4, p. 379–382. è. Ø. вðàôÂÚàôÜÚ²Ü, ¶. ¶. ÐàìвÜÜÆ êÚ²Ü, è. º. Øàôð²¸Ú²Ü, ø. ê. زð¶²ðÚ²Ü Üàð äàðüÆðÆÜܺðÆ ¶ºÜàîàøêÆÎ ¾üºÎîܺðÆ Ð²ØºØ²î²Î²Ü ²Ü²ÈƼÀ ¸ÜÂ-¶Æê²êîÔºðÆ ØºÂà¸àì IN VIVO ²Ù ÷ á ÷ á õÙ ¸ÜÂ-·Çë³ëïÕ»ñÇ Ù»Ãá¹áí ·Ý³Ñ³ïí»É ¿ Ýáñ åáñýÇñÇÝ` T OEt4PyP Ç ¨ Ýñ³ Ag-, Zn- ¨ Co-³Í³ÝóÛ³ÉÝ»ñÇ Ñ³Ù»Ù³ï³Ï³Ý ·»ÝáïáùëÇÏáõÃÛáõÝÁ ÙÏÝ»ñÇ ³ñÛ³Ý É»ÛÏáóÇïÝ»ñáõÙ in vivo ` ϳËí³Í ³½¹Ù³Ý ųٳݳÏÇó: ¸ÜÂ-Ç íݳëí³ÍùÝ»ñÇ Ù³Ï³ñ¹³ÏÇ μ³ñÓñ³óáõÙÁ áõëáõÙݳëÇñíáÕ åáñýÇñÇÝÝ»ñÇ Ý»ñ³ñÏáõÙÇó 24 ų٠³Ýó (Ý»ñ³ñÏáõÙÇó 6 ų٠³Ýó íݳëí³ÍùÝ»ñÇ Ù³Ï³ñ¹³ÏÇ Ñ³Ù»Ù³ïáõÃÛ³Ùμ) íϳÛáõÙ ¿ Ýñ³Ýó ÏáõÙáõÉÛ³ïÇí ¿ý»ÏïÇ Ù³ëÇÝ, ÇÝãÁ ϳñáÕ ¿ ÏÇñ³éí»É áõéáõóùÝ»ñÇ ùÇÙÇáûñ³ådzÛáõÙ: R. M. HAROUTYUNIAN, G. G. HOVHANNISYAN, R.Ye. MURADYAN, K. S. MARGARYAN COMPARAT IVE ANALYSIS OF GENOT OXIC EFFECT S OF NEW PORPHYRINS IN COMET ASSAY IN VIVO S u m ma r y Comparative genotoxicity of new porphyrins TOEt4PyP and their Ag-, Zn- and Co derivatives was estimated by Comet-assay in mice leukocytes in vivo, depending on time of treatment. The increase of level of DNA damage after 24 hours in comparison with 6 hours of porphyrins administration confirms their cumulative effect, that can be applied in chemotherapy of tumors. 155