Non periferal konumlarda florlu gruplar içeren tetra-sübstitüe ftalosiyaninlerin sentezi, fotofizikokimyasal ve biyolojik özelliklerinin incelenmesi
Synthesis and investigation of photophysicochemical and biological activities of tetra substituted phthalocyanines containing fluorinated moieties on non-peripheral positions
Yazar: ÇETİN ÇELİK
Danışman: PROF. DR. MAKBULE KOÇAK
İstanbul Teknik Üniversitesi / Lisansüstü Eğitim Enstitüsü / Kimya Ana Bilim Dalı / Kimya Bilim Dalı
Derece: Yüksek Lisans, 2021
Konu: Kimya = Chemistry

Özet:
Ftalosiyaninler sahip oldukları benzersiz fiziksel ve kimyasal özellikleri sebebiyle ilgi çeken makrohalka bileşiklerindendir. Spektrumun görünür bölgesinde kırmızıya yakın alanda yaptıkları güçlü absorbsiyon, delokalize olmuş 18 π-elekron sistemi, yüksek kimyasal ve termal kararlılık bu özelliklerden birkaçıdır. 600-800 nanometre arası yaptıkları güçlü absorbsiyon sebebiyle mavi-yeşil renge sahip olan ftalosiyaninler uzun yıllardır pigment olarak kullanılmakta olsalar da günümüzde kullanım alanları bununla sınırlı değildir. Kataliz, organik güneş hücreleri, sensörler, elektrokromizm gibi birçok alanda kullanılan ftalosiyaninler için özellikle ilgi çeken bir diğer kullanım alanı fotodinamik terapidir. Fotodinamik terapi, bir fotosensitizer aracılığıyla hücre içinde reaktif oksijen türleri oluşturarak hücre ölümünün sağlandığı, noninvazif bir kanser tedavi yöntemidir.
Ftalosiyaninlerin sahip oldukları özellikler, onları fotodinamik terapide kullanılmak üzere iyi bir fotosensitizer adayı yapmaktadır. Ancak zayıf çözünürlük, agregasyon yapma ve hidrofobik olma gibi özellikleri, ftalosiyaninlerin bu alanda kullanımını zorlaştırmaktadır. Ftalosiyanin halkasına sübstitüentlerin eklenmesi ve merkez metal iyonunun değiştirilmesi ile bu engellerin önüne geçilebilmektedir.
Bu tez kapsamında fotodinamik terapide kullanılabilme potansiyellerini, antioksidan ve antimikrobiyal özelliklerini incelemek amacıyla 7 adet yeni ftalosiyanin bileşiği  sentezlenmiştir. Bu doğrultuda önce 3-nitroftalonitril, 3,5-bis(triflorometil)fenol bileşiği ile kuru dimetil formamid içerisinde, azot atmosferi altında, oda sıcaklığında ve potasyum karbonat varlığında  48 saat boyunca reaksiyona sokulmuş, 3-(3,5-bis(triflorometil)fenoksi) ftalonitril (1) bileşiği elde edilmiştir. Elde edilen ftalonitrilin siklotetramerizasyon reaksiyonları sonucu hedeflenen ftalosiyanin bileşiklerinin sentezleri gerçekleştirilmiştir. Dimetilaminoetanol içerisinde, 135 ℃'de, metal tuzları (çinko(II)asetat, kobalt(II)klorür, bakır(II)asetat, nikel(II)klorür) kullanılarak metalli (3-6), metal tuzu kullanılmadan da metalsiz ftalosiyanin (2) türevleri sentezlenmiştir. Kinolin içerisinde, galyum(III)klorür varlığında galyum ftalosiyanin (7), 1-pentanol içerisinde, 1,8-diazabisiklo[5.4.0]undek-7-ene ve indiyum(III)klorür varlığında ise indiyum ftalosiyanin (8) sentezi gerçekleşmiştir. Yapılan tüm reaksiyonlar azot atmosferi altında gerçekleştirilmiştir.
Sentezlenen 3, 7 ve 8 bileşiklerinin farklı çözücüler içerisinde, ayrıca dimetilsülfoksit içerisinde farklı derişimlerdeki elektronik spektrumları, UV-vis spektrometresi aracılığıyla ölçülmüş, çözücünün cinsi ve derişimin spektoskopik ve agregasyon özellikleri üzerindeki etkisi araştırılmıştır. Bu bileşiklerin fotodinamik terapide kullanıma uyumlarını incelemek amacıyla, UV-vis ve floresans spekroskopisi kullanılarak, floresans kuantum verimleri, singlet oksijen kuantum verimleri ve fotobozunma kuantum verimleri hesaplanmıştır.
2, 3, 4, ve 5 bileşiklerinin 2,2-difenil-1-pikrilhidrazil serbest radikalini tutma ve demir (II) iyonu şelatlama aktiviteleri ölçülerek antioksidan özellikleri, disk difüzyon yöntemi kullanılarak da antimikrobiyal özellikleri incelenmiş, ayrıca test edilen bakterilerin, yüksek antimikrobiyal aktivite gösteren 5 bileşiğine karşı renk açma etkileri araştırılmış ve UV-vis absorbans değerleri deneyin başında, 2, 5 ve 10 gün sonra gösterilmiştir. 5 bileşiğinin kullanıldığı bütün bakteri suşlarının 10 günün sonunda boyayı bozduğu bulunmuştur. Ancak 5 bileşiğinin metil laurat ile karıştıldığında bu etkinin yavaşladığı gözlenmiştir.
Sonuç olarak bu çalışmada bir adet ftalonitril ve bu ftalonitrilden yedi adet yeni ftalosiyanin bileşiği sentezlenmiştir. Sentezlenen bileşiklerin karakterizasyonları FT-IR, UV-vis, NMR, ve MALDI-TOF teknikleri kullanılarak gerçekleştirilmiştir. Karakterizasyonu gerçekleştirilen  ftalosiyaninlerin bir kısmının floresans kuantum verimleri, singlet oksijen kuantum verimleri ve fotobozunma kuantum verimleri hesaplanmış, bir kısmının ise antioksidan ve antimikrobiyal özellikleri incelenmiş, ilave olarak 5 bileşiğinin bakteri ortamında renksizleştirme çalışmaları yapılmıştır.

Abstract:
Phthalocyanines are a group of rather interesting macrocyclic compounds due to their unique physical and chemical properties such as strong absorption in the near red region of visible spectrum, delocalized 18 π-electron system, high chemical and thermal stability. Although phthalocyanines, which have a blue-green colour due to their strong absorption between 600-800 nm wavelength, have been used as pigments for many years, their usage areas are not limited to this. Some areas of interest for phthalocyanines can be listed as catalysis, organic solar cells, sensors, electrochromism and photodynamic therapy. Antioxidant and antimicrobial activities of various phthalocyanine compounds have also been investigated.
Photodynamic therapy is a non-invasive cancer treatment method in which cell death is achieved by creating reactive oxygen species inside the cell through a photosensitiser. The properties of phthalocyanines make them good candidates as photosensitisers for use in photodynamic therapy. However, their poor solubility, tendency to aggregate and hydrophobicity cause difficulties for their usage in this field. Nevertheless, these problems can be avoided by adding substituents to the phthalocyanine ring and replacing the central metal ion.
Phthalocyanines which are substituted by electron donating groups at their non-peripheral positions show higher red shift than that of peripherally substituted ones. This makes non-peripherally substituted phthalocyanines better candidates to be used in photodynamic therapy than their peripherally substituted counterparts. Diamagnetic metal ions show considerably more singlet oxygen quantum yield, which is a very important parameter for photosensitisers. Therefore phthalocyanines containing metal ions such as Zn+2, Ga+3 and In+3 in their core are preferred for photodynamic therapy applications.
Fluorinated phthalocyanines possess certain advantages over their non-fluorinated counterparts. It has been shown that fluorinated phthalocyanines display increased lipophilicity, higher solubility and less aggregation. These properties make phthalocyanines that has fluorinated groups as substituents good candidates in photodynamic therapy. 
With increasing drug resistance resulting from the broad use of antibiotic drugs, the search for new antimicrobial agents has gained importance. Phthalocyanines are a group of molecules that has shown good antimicrobial activity. Phthalocyanine compounds are very much available in many different ink formulations as colorants. It has been shown that different bacteria cause the biodegradation of  phthalocyanines. Phthalocyanine compounds with varying properties have been the subject of antimicrobial activity studies. However, there has not been any studies investigating the synergistic effect of phthalocyanines and other additives used in ink formulations.
This study aims to synthesize seven new phthalocyanine compounds and study their biological features including their potentials as photosensitisers in photodynamic therapy and their antioxidant and antimicrobial activity. To achieve this, first 3-nitrophthalonitrile was first reacted with 3,5-bis(trifluoromethyl)phenol in dry dimethyl formamide, under nitrogen atmosphere, at room temperature and in the presence of potassium carbonate for 48 hours and 3-(3,5-bis(trifluoromethyl)phenoxy) phthalonitrile (1) compound was obtained. The targeted phthalocyanine compounds were synthesized by the cyclotetramerisation reactions of the obtained phthalonitrile. Metallophthalocyanines (3-6) were synthesised using related metal salts (zinc(II)acetate, cobalt(II)chloride, copper(II)acetate, nickel(II)chloride) in dimethylaminoethanol, at 135 ℃. According to the same procedure, metal-free phthalocyanine (2) was obtained in the absence of metal salt. Synthesis of gallium phthalocyanine (7) was carried out in quinoline with the presence of gallium(III)chloride, while the synthesis of indium phthalocyanine (8) was done in 1-pentanol with the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene and indium(III)chloride. All reactions were carried out under nitrogen atmosphere.
Electronic spectra of the synthesized compounds 3, 7 and 8 in different solvents and also in dimethylsulfoxide at different concentrations were measured by UV-vis spectrometer, and the effects of solvent type and concentration on their spectroscopic and aggregation properties were investigated. All three compounds showed good solubility in both polar and apolar solvents. In dimethylsulfoxide all three compounds showed linear increase in absorbtion with respect to concentration. In order to examine the capacity of these compounds for potential use in photodynamic therapy, fluorescence quantum yields, singlet oxygen quantum yields and photodegradation quantum yields were calculated using UV-vis and fluorescence spectroscopy data. The comparative method against unsubstituted zinc phthalocyanine was used for the calculations. All three compounds showed similar fluorescence quantum yields to that of unsubstituted zinc phthalocyanine, moderate photostability and singlet oxygen quantum yields. 
The antioxidant properties of 2, 3, 4, and 5 compounds were observed by measuring their scavenging ability of the free radical 2,2-diphenyl-1-picrylhydrazyl as well as their ferrous ion chelating activity, and their antimicrobial properties were investigated using disc diffusion method. 4 showed the highest antioxidant activity. 2 and 5 showed similar antioxidant activity relative to the each other while 3 showed the least. When the synergistic effects of methyl laurate and phthalocyanines were investigated, it was found that they showed moderate synergistic effects for radical scavenging activities. In disc diffusion assay, against the four different strands of bacteria, 5 and 4 showed the highest antimicrobial activity. When the synergistic effects were investigated by mixing methyl laurate and phthalocyanines, it was observed that the inhibition zones increased substantially aside from 3 which showed the lowest antimicrobial activity. 
For compound 5 and its mixture with methyl laurate, which showed the highest antimicrobial activity, decolourisation effect of bacteria was investigated by measuring the decrease in UV-Vis absorbance at the start, 2, 5 and 10 days later. It was found out that all bacteria strands used were able to degradate compound 5 almost completely in 10 days. However, it was observed that mixing methyl laurate slowed the degradation process considerably. 
In this study, a phthalonitrile compound and seven novel phthalocyanine compounds were synthesized. Characterization of synthesised compounds were carried out using FT-IR, UV-vis, NMR, and MALDI-TOF techniques. Fluorescence quantum yields, singlet oxygen quantum yields, photodegradation quantum yields, antioxidant and antimicrobial properties of the characterised phthalocyanines were investigated. Additionally, decolourisation studies of compound 5 in bacteria media were carried out.