Non-canonical anti-cancer, anti-metastatic, anti-angiogenic and immunomodulatory PDT potentials of water soluble phthalocyanine derivatives with imidazole groups and their intracellular mechanism of action.

Cancer is currently a leading health issue globally. Chemotherapy is a prominent treatment method but due to undesired side effects t, there has been a need for novel less toxic approaches. Photodynamic therapy may be listed among the alternatives for efficient and potentially less detrimental applications of cancer therapy. Canonical photodynamic therapy (PDT) approach requires a light source with a specific wavelength of light, a non-toxic photosensitizer and molecular oxygen. PDT creates the desired effect by the photochemical reaction created through interaction of these components to create reactive oxygen species that will act on the cancer cells to enable anti-cancer activities. In our study we focus on non-canonical PDT application. In this approach we are not only aiming to eliminate cancer cells in the environment but also test the anti-metastatic, anti-angiogenic and possible immunomodulatory activities of the novel photosensitizers. Moreover, in our approach, we studied the intracellular pathways that are crucial for carcinogenesis, cell cycle, apoptosis, angiogenesis, metastasis and immune function to decipher the mechanism of the action for each compound. Reactive oxygen species based explanation was not valid in our study, hence it brings out a non canonical approach to PDT applications. Our results suggests that Phthalocyanine derivatives with imidazole groups can be effectively used against lung, colon, breast and prostate cancer while differentially effecting metastasis, angiogenesis, cell cycle, apoptosis and immune system cells' activities. Based on the results, PDT application of these phthalocyanine derivatives can be an effective treatment option to replace chemotherapy to minimize the potential side effects.

Imaging and detection of cell apoptosis byIn vitrophotodynamic therapy applications of zinc (II) phthalocyanine on human melanoma cancer.

This study aims to investigate the photodynamic therapy (PDT) effects on MeWo (human melanoma cells) and HaCaT (normal human keratinocyte cells) by light stimulation of different concentrations of Zinc (II)-tetra-tert-butyl-phthalocyaninato (ZnPc). MTT viability assay data indicated that a 25 µM concentration of ZnPc is cytotoxic to the melanoma cancer cells while this concentration of ZnPc is not cytotoxic for the HaCaT cell line. Moreover, the results showed that photoactivated ZnPc at 12.5 µM concentration reduced the cell viability of the MeWo cell line to about 50 %. At this photosensitizing concentration, the efficacy of light doses of 20, 30, 40, and 50 J/cm2 was evaluated against MeWo and HaCaT cells. ZnPc at a concentration of 12.5 µM activated with a light dose of 50 J/cm2 was the most efficient for the killing of MeWo cells. In conclusion, the 12.5 µM of ZnPc with the treatment light dose of 50 J/cm2 from a RED light source was adequate to destroy MeWo cells by the ROS-induced apoptosis mechanism. It also exhibited low killing effects on healthy HaCaT cells. These findings are supported by the results of apoptosis with the Annexin V & Dead Cell Kit and fluorescence imaging.

Detection of Kallikrein-Related Peptidase 4 with a Label-free Electrochemical Impedance Biosensor Based on a Zinc(II) Phthalocyanine Tetracarboxylic Acid-Functionalized Disposable Indium Tin Oxide Electrode.

A new impedimetric biosensing system based on kallikrein-related peptidase 4 (KLK 4) antigen-specific antibodies and a zinc(II) phthalocyanine tetracarboxylic acid (Zn-PcTCa) matrix material was developed for the first time in this study. First, a Zn-PcTCa-coated indium tin oxide surface was used as an interface matrix material for the immobilization of anti-KLK 4 antibodies, and they bound to the platform via amide bonds. In the presence of KLK 4 antigens, the anti-KLK 4 antibodies specifically captured these antigens and caused changes in the electrochemical properties of the system. Randles equivalent circuit was utilized to evaluate the impedimetric signal, which was measured with the help of an electrochemical impedance spectroscopy method. After the specific interaction, the electron transfer resistance (Rct) was remarkably increased and displayed a linear relationship with the level of the KLK 4 antigen in the range of 0.02-15 pg/mL, with a a detection limit of 6.8 fg/mL. The designed biosensor was able to detect a KLK 4 antigen with good sensitivity, excellent specificity, and high stability. In addition, because of having a low-cost and robust procedure for fabrication, it could be repeatedly used in several areas including clinical diagnosis.

Photo induced anti-inflammatory activities of a Thiophene substituted subphthalocyanine derivative.

Materials that possess photo induced biological activities present opportunities for more localized, targeted and efficient treatment options that may also reduce side effects. There have been studies supporting photo induced effects of photosensitizers as anti-cancer, anti-inflammatory and anti-microbial agents. In this study, we tested photo activated anti-inflammatory effects of a Thiophene substituted subphthalocyanine (SubPc) derivative. Thiophene and its derivatives are well known for their anti-inflammatory and anti-microbial effects. There are unwanted side effects associated with Thiophene derivatives. By substituting this biologically active molecule to SubPc structure we acquired control over its activation. Upon light treatment this derivative exerted anti-inflammatory activity on the mammalian macrophages in vitro based on the substantial decrease in extracellular inflammatory cytokine levels. Our results suggest that Thiophene substituted SubPc derivative has photo induced anti-inflammatory activities. This material can be used for the treatment of patients suffering from chronic inflammation that are not associated with a bacterial burden such as autoimmune diseases and inflammatory or allergic reactions.

Anti-Cancer and Anti-Inflammatory Activities of Bromo- and Cyano-Substituted Azulene Derivatives.

Natural products and their synthetic derivatives gathered attention due to their pharmaceutical capacities. They have been in use against different types of diseases ranging from cancer to inflammatory disorders. In order to increase their efficacy and prevent the possible side effects, these natural compounds are manipulated at the laboratory conditions and modified according to our needs. Azulene is one of these compounds whose anti-inflammatory potential have been shown by the previous studies, but a detailed analysis of its effect at the cellular level in terms of pro-inflammatory cytokine production has not been studied yet. Moreover, its derivative potential has not been characterized extensively. In our study, we examined the cytotoxic, immunomodulatory and immunostimulatory potential of bromo- and cyano-substituted azulenes on the mammalian macrophages. These unique compounds had differential effects on the production of pro-inflammatory cytokines and they were anti-inflammatory immunomodulators. Furthermore, they exerted anti-proliferative effect on breast and prostate cancer cells which supports their anti-cancer potential as well.

Immunomodulatory activities of zinc(II)phthalocyanine on the mammalian macrophages through p38 pathway: Potential ex vivo immunomodulatory PDT reagents.

A series of zinc phthalocyanine having imidazolyl moieties was synthesized. These compounds' immunostimulatory and immunomodulatory activities were tested on the mammalian macrophages in vitro. In the absence of photo induction neither dmso soluble nor the water soluble imidazole Pc had any immunostimulatory or immunomodulatory effect on the macrophage activity based on the differences in the pro-inflammatory cytokine secretion levels compared to the control groups. Upon photo induction, especially, at 5 min exposure time both derivatives lead to an increased pro-inflammatory cytokine secretion level by LPS activated macrophages. Whereas, this effect was completely reversed after 10 min of light treatment and both derivatives gained stark anti-inflammatory potential. Our molecules were cell penetrating and exerted their effects by regulating the phosphorylation levels of p38. This study is one of its first examples suggesting differential immunomodulatory photo dynamic therapy applications of phthalocyanine derivatives depending on light exposure time as well as their possible route of modulating the intracellular signaling pathways.

Immunostimulatory effect of Zinc Phthalocyanine derivatives on macrophages based on the pro-inflammatory TNFα and IL1ß cytokine production levels.

In this study, we examined the effect of halogen substitution on the ability of ZnPcs to stimulate or regulate the immune system cells. There have been studies focusing on the usage of Pcs as treatment option against different cancer types. More attention should be paid on their possible positive or negative effects on the immune system for better prognosis rates. We designed and synthesized unique ZnPcs with iodine substitution and further tested their effect on mammalian macrophage cell line. Macrophages are crucial cell types that can define the immune response by cytokine production as well as by antigen presentation to the other immune system cells after phagocytosis of the danger associated molecules. Our results suggest an immunostimulatory role for the iodine substituted ZnPc on macrophages based on the changes in pro-inflammatory cytokine production levels (TNFα, IL1ß and IL6). This effect dependent on the presence of iodine; since non-substituted ZnPc did not exert such a stimulatory activity on the macrophages. These results support a possible use of the reagents against the tumor types that would get affected detrimentally by the pro-inflammatory environment.

Photophysicochemical properties and photodynamic therapy activity of highly water-soluble Zn(II) phthalocyanines.

The syntheses of two zinc(II) phthalocyanines (ZnPcs) having either imidazole (ZnPc 1) or pyridiloxy (ZnPc 2) moieties as their macrocycle substituents are reported. Quaternization of the ZnPcs with methyl iodide afforded water soluble cationic phthalocyanines. The photophysical, photochemical properties and photodynamic therapy (PDT) activity of the ZnPcs were studied in solution. The fluorescence quantum yield and lifetime of ZnPc 1 were higher as compared to ZnPc 2. ZnPc 2 afforded higher triplet state (ΦT) and singlet oxygen quantum yields (ΦΔ) in comparison to ZnPc 1. The PDT activity of ZnPcs was investigated against human breast adenocarcinoma cells (MCF-7). The two compounds afforded a very minimal in vitro dark cytotoxicity with 85% viable cells at concentration ≤80 µM. On irradiation of the cells having the ZnPcs, ≥50% cell death was recorded for ZnPc 1 which was also evidenced by the cells photo-micrograph.

Construction and amperometric biosensing performance of a novel platform containing carbon nanotubes-zinc phthalocyanine and a conducting polymer.

A novel glucose oxidase (GOx) based amperometric biosensor utilizing a conducting polymer (CP), multi walled carbon nanotubes (MWCNTs) and a novel water soluble zinc phthalocyanine (ZnPc) was constructed. For this purpose, a novel ZnPc was synthesized to examine the role of being a part of support material for enzyme deposition. High water solubility was achieved with the introduction of tetra quaternized imidazolyl moieties at the peripheral positions of phthalocyanine. In order to fabricate the proposed biosensor, a graphite electrode was firstly modified with poly[9,9-di-(2-ethylhexyl)- fluorenyl-2,7-diyl] end capped with N,N-Bis(4- methylphenyl)-4-aniline (PFLA) and MWCNTs. Then, GOx was co-immobilized with ZnPc onto the modified surface. To the best our knowledge, a sensor design which combines conjugated polymer/MWCNTs/ZnPc was attempted for the first time and this approach resulted in improved biosensor characteristics. The constructed biosensor showed a linear response for glucose between 0.025-1.0mM with a detection limit of 0.018mM. KMapp and sensitivity values were calculated as 0.53mM and 82.18µAmm-1cm-2, respectively. Moreover, scanning electron microscopy (SEM) and cyclic voltammetry (CV) techniques were used to investigate the surface modifications. Finally, fabricated biosensor was tested on beverages for glucose detection successfully.