Photosensitive Antitumor Activity Of A Series Of Polyamine Phthalocyanines And Their Water-soluble Derivatives

Photodynamic therapy(photodynamic therapy,PDT)means tumor tissue under the effect of photosensitizer and light together,the organism cells or biological molecules change their morphological or functional form,in serious cases lead to cell damage and necrosis.This is a new therapie of being studied and rapid developing,which has become one of the most active research areas in the World Cancer Prevention science.Phthalocyanine complexes have advantages of high efficiency and low toxicity has become the focus of anticancer drugs as the photosensitizers.This article selects two series of multi-amine phthalocyanines and its corresponding different ion type phthalocyanine derivatives as research subjects.2(3),9(10),16(17),23(24)-tetra-(((2-(trityl-amino)ethylamino)methyl)phenoxy)phthalocyanine zinc(Ⅱ)(ZnPc1);2(3),9(10),16(17),23(24)-tetra-(((2-amino-ethyl-amino)methyl)phenoxy)phthalocyanine zinc(Ⅱ)(ZnPc2);quaternized 2(3),9(10),16(17),23(24)-tetra-(((2-amino-ethyl-amino)methyl)phenoxy)phthalocyanine zinc(Ⅱ)(ZnPc3);2(3),9(10),16(17),23(24)-tetra-(((2-(diethylamino)ethylamino)methyl)phenoxy)phthalocyanine zinc(Ⅱ)(ZnPc4);quaternary amination of 2(3),9(10),16(17),23(24)-tetra-(((2-(diethylamino)ethylamino)methyl)phenoxy)phthalocyanine phthalocyanine zinc(Ⅱ)(ZnPc5).This article is based on the structural characteristics of the phthalocyanine,classified ZnPc1、ZnPc2 and ZnPc3 as the series one,classified ZnPc4 and ZnPc5 as the series two.First,compare the photophysical and photochemical properties of the multi-amine phthalocyanine and its corresponding different ion type phthalocyanine derivatives.Secondly,detecting phthalocyanines have effects on tumor cell proliferation by MTT assay;it confirmed that phthalocyanines have effect of inhibiting the proliferation of Hela and SGC-7901 cells in a concentration range of 10～25 μgmol/L,by filtering appropriate drug dose and finaly selected 5 μmol/L concentration range as the experimental dose.Finally,studied the anti-tumor activity of the phthalocyanine photosensitizers,nuclear chromatin changes and generation of reactive oxygen species generation(ROS)levels in intracellular,which provided further study of containing the the polyamine structure of zinc phthalocyanine in photodynamic therapy as foundation.The main findings are as follows:(1)A hydrogen atom on the phthalocyanine ring is substituted by a quaternary ammol/Lonium base group,the series one:the singlet oxygen generating capacity relationship of phthalocyanine:ZnPc3>ZnPcl>ZnPc2;the relationship of the reactive oxygen species level generated within the cells:ZnPc3>ZnPc1>ZnPc2;the relationship of the inhibition ability of tumor cells:ZnPc3>ZnPc1>ZnPc2.Series two:the singlet oxygen generating capacity relationship of phthalocyanine:ZnPc5>ZnPc4;the relationship of the reactive oxygen species level generated within the cells:ZnPc5>ZnPc4;the relationship of the inhibition ability of tumor cells:ZnPc5>ZnPc4.Containing quaternary ammol/Lonium salt water-soluble phthalocyanine singlet oxygen generation capability significantly stronger than non-quaternized phthalocyanine,and the intracellular level of reactive oxygen species generation capacity and the capacity of photosensitive anti-tumor activity consistent with singlet oxygen generating ability level.(2)Series one:the lipid-water partition coefficient of phthalocyanines:ZnPc1>ZnPc2>ZnPc3;the zeta potential of phthalocyanine solution:ZnPc3>ZnPc2>ZnPcl;the drug uptake rate:ZnPc3>ZnPc2>ZnPcl.Series two:the lipid-water partition coefficient of phthalocyanines:ZnPc4>ZnPc5;the zeta potential of phthalocyanine solution:ZnPc5>ZnPc4;the drug uptake rate:ZnPc5>ZnPc4.The smaller is lipid-water partition coefficient,the more positively electricity of phthalocyanine,the higher the rate of uptake of the drugs on the cells.Because the positively charged of containing quaternary ammol/Lonium salt water-soluble phthalocyanine solution,and the cell solution was negatively charged,polyamine phthalocyanine with the positive charge of the N+ by inducing action and will produce a strong electrostatic interaction between the negatively charged cells,it’s conducive to the absorption of the cells to drugs.(3)The studies of phthalocyanine reactive oxygen species in the case by ADPA light oxidation bleaching determination of singlet oxygen and DCFH-DA probe to detect intracellular reactive oxygen species.The results show that:the generation of singlet oxygen of water-soluble quaternary ammonium salt derivatives phthalocyanine was stronger than the non-quaternized phthalocyanine,and positively correlated with the level of intracellular reactive oxygen species.Mainly in:(1)the relationship of singlet oxygen generation speed:ZnPc3(k=0.1111)>ZnPc1(k=0.01967)>ZnPc2(k=0.00948);ZnPc5(k=0.03934)>ZnPc4(k=0.02767);(2)the relationship of intracellular level of reactive oxygen species(ROS)generation:ZnPc3>ZnPc1>ZnPc2;ZnPc5>ZnPc4.(4)By MTT assay studies showed that five kinds of phthalocyanine have photosensitive anti-tumor activity.Mainly in:(1)resulting change in tumor cell morphology and nuclear chromatin damage;(2)the relationship of the photosensitive inhibition ability to tumor cells:ZnPc3>ZnPc1>ZnPc2;ZnPc5>ZnPc4.The above experimental results show that the five kind amine phthalocyanine and water-soluble derivatives have good safety and photosensitive antitumor activity ability in this paper.The anti-tumor activity ability of drugs was positively correlated with intracellular reactive oxygen species and singlet oxygen.Quaternary ammonium derivatives of phthalocyanine having a higher Zeta potential,a smaller lipid-water distribution coefficient and high tumor uptake,and thus has a better photodynamic activity.Which provide a guide to design and synthesis new structure phthalocyanine that have good photosensitivity,but also for further promote the phthalocyanine photosensitizer in the the PDT field of clinical application process.