Suda çözünür yeni makrohalkalı bileşiklerin sentezi ve biyolojik özelliklerinin incelenmesi
Synthesis of water-soluble new macrocyclic compounds and investigation of their biological properties
Yazar: AYNUR KÜBRA MUT
Danışman: DOÇ. DR. BEHİCE ŞEBNEM SESALAN
İstanbul Teknik Üniversitesi / Lisansüstü Eğitim Enstitüsü / Kimya Ana Bilim Dalı / Anorganik Kimya Bilim Dalı
Derece: Yüksek Lisans, 2022
Konu: Kimya = Chemistry
Dizin: Floresan = Fluorescence ; Ftalosiyaninler = Phthalocyanines ; Spektrofotometri-ultraviyole = Spectrophotometry-ultraviolet ; Termal denatürasyon = Thermal denaturation

Özet:
Geçtiğimiz pandemi sürecinde DNA, mRNA kavramları hayatımıza çok hızlı ve kalıcı şekilde girmiştir. Bunun sebebi koronavirüsün hayatımızı tehdit eder boyutta hastalığa sebep olmasıdır. Tedavi amaçlı mRNA aşılarının kullanılması sadece DNA değil RNA üzerine odaklanmamıza sebep olmuştur. 
Pandemi sürecinde koronavirüse karşı ilaç geliştirilmesi çok büyük önem kazanmıştır. Bu noktada bildiğimiz şey şudur ki vücudumuzda enfeksiyona sebep olan bir bakteri veya virüsün vücut içinde kendi DNA'sının kopyalanmasını durdurduğumuz zaman virüsün  hücre çoğalması da  önlenmiş olur. DNA replikasyonunun durdurulmasının bir yolu DNA bazlarının tanınmaz hale gelmesini sağlamaktır.
Bu amaçla genel olarak ilaç tasarımında ilaçların etken maddelerinin DNA ile kompleks oluşturması beklenmektedir. Kimyasal maddeler DNA'ya bağlandıkları zaman replikasyon işlemi durmaktadır. Çünkü DNA enzimleri ilaca bağlanan bazları tanımaz.
Etkin ilaç üretmek için o etken maddenin veya kimyasal maddelerin özelliklerinin bilinmesi kritik bir noktadır. Kimyasalın toksik özelliği varsa daha çok kansere yönelik çalışmalarda kullanılabilir fakat bu toksisite sağlıklı hücreye zarar veriyorsa o ilaç etkin şekilde kullanılamaz. İşte tam bu noktada daha biyo-uyumlu ilaçlar ihtiyaç haline gelmiştir.
Son yıllarda biyomedikal alanda ikinci-nesil porfirin türevleri olan ftalosiyaninlere (Pc) daha fazla odaklanılmıştır. Sadece kanser tedavide değil antibakterial-antimikotik olarak, retina kalınlaşmasını tedavide, histopatolojide canlı ve ölü hücreyi birbirinden ayırmada Pc bileşikleri kullanılmaktadır. Pc bileşiklerinin özellikle DNA sarmalı ile etkileşim gösterdiği bilinmektedir. Pc türevlerinin her biri DNA ile farklı şekilde etkileşime girmektedir.  Metalli veya metalsiz periferal veya non-periferal sübstitüentli düzlemsel yapıda olan Pc'ler çoğunlukla interkalasyon yapabilmektedir. Fakat silisyum Pc gibi eksenel konumlara bağlı grupları olan Pc türevleri daha hacimli olduğundan DNA bazları veya fosfat-şeker omurgasıyla elektrostatik etkileşime girebilmektedir. Yeni sentezlenen Pc türevlerinin DNA ile etkileşime girip girmediğinin tespit edilmesi için ilk adım etkileşimin UV-vis ve floresans spektrumları ile gözlenmesidir.
İster antibakteriyel ister antikanserojen isterse antiromatizmal ilaç olsun biyoteknolojik alanda etken maddenin DNA ile etkileşiminin olup olmaması en kritik noktadır. 
Bu amaçla bu tez kapsamında yeni suda çözünür asimetrik Pc bileşiklerinin sentezi tasarlanmış ve elde edilenlerin DNA etkileşimleri UV-vis, floresans spektroskopileriyle incelenmiştir. DNA termal denaturasyon grafisinde DNA erime noktası azalttığı yani heliks sarmalının yeni Pc bileşiğiyle etkileşime girerek daha düşük sıcaklıklarda bozunduğu görülmüştür.

Abstract:
Recently, due to pandemic situation the concepts of DNA and RNA has contributed in our lives so fast because coronavirus caused so dangerous diseases that effect our whole lives. So we focused on not only DNA but also RNA by mRNA vaccines. During pandemic situation, the development of drugs for the treatment of coronovirus became very important for all the world. At that point, it is known that to stop the cell division of virus or bacteria in our body that caused infection, the replication of the viral DNA should be stopped. One of the ways to do that is the prevention of recognition of the bases on DNA.
For this purpose, generally, the complex formation between DNA and active components of drugs is expected during the design of new drugs. When whether active components of drugs or chemical compounds bound to bases on DNA, replication stops because the enzymes that proceed the replication does not recognize the bases which bound to drug.
To produce an active efficient drug, the properties of this active compound should be known. If it is highly toxic, then this compound can be used for cancer treatment. However, this toxicity may have adverse effects to healthy cells in the body. At that point, more biocompatible drug design is requred.
Recently, the developments in biomedical area focused on the second generation porphyrin derivatives; phthalocyanines (Pcs). The main phthalocyanine compound is synthetic macrocyclic structures with chemical formulas such as C32H18N8 or (C8H4N2)4H2, obeying Hückel's rule, with a system of 18 π-electrons surrounded by 2 hydrogen atoms in the core center and 16 hydrogens. They are also called 'tetrabenzotetraazaporphyrin' because they are the condensation product of four iminoisoindoline units.
Phthalocyanines are synthetic substances that have similar structures to porphyrins such as hemoglobin, chlorophyll a and vitamin B12, but are not found naturally like porphyrins. As the most general method to make phthalocyanines soluble, substituted phthalocyanines are obtained by adding functional groups such as amide, carboxylic acid, tertiary butyl units from peripheral or non-peripheral positions.
Within the scope of this experiment, 8 different purification methods were used. Of these, 2 (Thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC), column chromatography on silica gel, solvent evaporation or recrystallization ) were used much more frequently.
One of the most defining features of phthalocyanines is their color; It changes from red to blue and from blue to green. It has been observed that phthalocyanines containing alkali and alkaline earth metals do not sublimate at high temperatures and under vacuum.
Metal-free phthalocyanines are planar and have D2h symmetry. Metallic phthalocyanine molecules are in D4h symmetry, and by axial bonding of various molecules to the metal, the square planar structure turns into a five-coordinated pyramidal structure or a six-coordinated octahedral structure.
Phthalocyanine molecules are aromatic because they have 16 atoms and 18π-electrons according to Huckel's Rule.
The characteristic and defining absorption spectra of phthalocyanines are the Soret (B) band at 350 nm and the strong Q band at 675 nm.
The Q-band absorptions of red-shifted phthalocyanines compared to porphyrins are approximately twice as large as the Q-band absorptions of porphyrins, and the reason for this shift to different wavelengths is that the absorption spectrum of phthalocyanine molecules shifts to longer wavelengths, and the pyrrole groups conjugated with benzene rings are bonded to each other with nitrogen atoms.
Phthalocyanines are easily oxidized and reduced. Oxidation and reduction can occur in the metal atom or in the ring.
During the synthesis of phthalocyanines, which have a highly tense structure and consist of four iminoisoindole nuclei, the efficiency of the reactions in the production of metal phthalocyanines is higher than the metal-free phthalocyanines, due to the metal ion directing (template) effect in the environment. Aromatic o-dicarboxylic acids or nitrile, imide and amide derivatives of these acids can be used for the synthesis of phthalocyanines.
Metal-containing phthalocyanines can be collected in two parts as electrovalent and covalent. Electrovalent phthalocyanines generally contain alkali and alkaline earth metal phthalocyanines and do not show solubility in organic solvents. When it interacts with dilute inorganic acids, aqueous alcohol, and even water, the metal ion is separated from the molecule and metal-free phthalocyanine is obtained.
Phthalocyanines catalyze many important chemical reactions, thanks to the redox active metal ions they contain in their center. It is also used as a catalyst in oxidation reactions, homogeneous and heterogeneous catalysis systems. It is widely used as an effective catalyst in the anodic oxidation of sulfur dioxide, cathodic reduction of nitric oxide, carbon dioxide and oxygen in electrochemical processes. Metallic phthalocyanines, which are used as solid catalysts in reactions, are preferred because of their ease of recycling and recovery.
The molecule with the diameter of the metal ion corresponding to the diameter of the central cavity of the molecule shows a stable structure. If the diameter of the metal ion is larger or smaller than the cavity diameter of the phthalocyanine, there is no stability and in this case the metal ions are easily separated.
Metallic, neutral, etc. structural properties of phthalocyanines, the formation of peripheral substituents and the position of the substituted group, the size of phthalocyanines are very effective on aggregation, and this aggregation changes the physical and chemical properties of phthalocyanines.
A good NMR spectrum may not be obtained in terms of phthalocyanine aggregation that may occur in the solution, causing broadening of the peaks, and the peak positions may change drastically as a result of this aggregation. The peaks obtained in the spectrum can be improved by choosing high temperature, low concentration or suitable solvent.
Asymmetric or low symmetry phthalocyanines are named this way because of the different substituents in their peripheral positions.
Asymmetric phthalocyanines containing two different isoindole units (A and B) are synthesized according to the type and shape of the targeted product ( A3B, A2 B2, AB3).There are different methods such as statistical condensation method and subphthalocyanine approach for the synthesis of an asymmetric phthalocyanine type A3B , the most widely used synthesis method .
Silicon phthalocyanines (SiPc), which have a silicon atom in the center, have low aggregation tendencies and have high singlet oxygen yields, allow their use as a DNA-targeted tool as well as being used in photochemotherapy.
High quantum yields were observed in sulfonated silicon phthalocyanines. This feature makes them suitable candidates for PDT.
Silicon phthalocyanines can be synthesized with diiminoisoindoline, anhydride, dinitrile and metal-free phthalocyanines by different methods [52].
Phthalocyanines, with their colors ranging from blue to green, are widely preferred in textile, inkjet, fountain pen inks, coloring plastic and metal surfaces, and in the paper industry used as thin films for coating material layers used in semiconductor units and optical devices. At the same time, phthalocyanines with electrochromic properties are also used in the production of self-darkening mirrors, display panels, the control of heat and light passing through glass materials, and the production of smart materials.
These macrocyclic compounds not only for cancer treatment but also are used as antimicotic-antibacterial agents. They are used in treatment of macula and to discriminate the alive cell in histopathology. It is known that Pcs reacts with DNA helix. While metallic or non-metallic peripherally or non-peripherally substituted planar Pcs interact with DNA intercalatively, more bulky structure such as silicon Pcs bind with phosphate-sugar backbone of DNA electrostatically. The first step to test the new Pcs compounds whether they bind with DNA or not is to observe the interaction by the use of UV-vis and florescence spectra. 
Whether the drug is syhthesized as anticancer or antibacterial-antiviral or antiromatoid, the critical point is to see its interaction with DNA.
For this purpose, within the scope of this thesis, based on the experimental information in the literature, it was tried to synthesize new asymmetric metal Pcs, which are predicted to have the potential to interact with DNA by making them cationic. A new axially substituted silicon Pc ( 8 ) was synthesized, and then this compound was quaternized and a new water-soluble product was obtained by activating the plan B, since the synthesis was negative . Item 1 was characterized by elemental analysis with H-NMR, mass, UV-Vis spectra. The interactions of this compound with DNA were investigated by UV-Vis, fluorescence spectra and thermal denaturation graph, and it was thought that the water-soluble compound 9 had electrostatic interaction with DNA.
Keywords: DNA, phthalocyanine, UV-Vis-spectrophotometry, thermal denaturation, fluorescence spectrophotometry.