Water-soluble silicon phthalocyanine containing a pyridine group with high activity in lung cancer cell line

Abstract

The invention relates to silicon phthalocyanine compounds containing pyridine groups with anti-cancer (cancer-therapeutic) and antioxidant effects, which exhibit activity in cancer cells that show resistance to metal-containing compounds, have a broader activity spectrum compared to metal-containing compounds, possess lower toxicity and reduced side effects compared to metal-containing compounds, and can be used as potential anti-tumor drugs, as well as to the methods of synthesis of these compounds.

Description

[0001] DESCRIPTION WATER-SOLUBLE SILICON PHTHALOCYANINE CONTAINING A PYRIDINE GROUP WITH HIGH ACTIVITY IN LUNG CANCER CELL LINE

[0002] Technical Field

[0003] The invention relates to silicon phthalocyanine compounds containing pyridine groups with anti-cancer (cancer-therapeutic) and antioxidant effects, which exhibit activity in cancer cells that show resistance to metal-containing compounds, have a broader activity spectrum compared to metal-containing compounds, possess lower toxicity and reduced side effects compared to metal-containing compounds, and can be used as potential anti-tumor drugs, as well as to the methods of synthesis of these compounds.

[0004] State of the Art

[0005] According to the latest report published by the World Health Organization, cancer is among the top three leading causes of premature death before ages 30-69 in 177 out of 183 countries. According to 2022 data, approximately 20 million new cancer cases were reported, and about 10 million people died due to the disease. In general, cancer incidence and mortality are rapidly increasing worldwide, which is attributed to both the aging and the growth of the population. There is also an increasing prevalence of the main risk factors for cancer, many of which are associated with socioeconomic development.

[0006] Lung cancer is a type of cancer characterized by the uncontrolled proliferation and development of lung cells and the appearance of neoplasms within the lung. With an estimated 2.5 million new cancer cases and 1.8 million deaths, lung cancer is the third most commonly diagnosed cancer and the leading cause of cancer deaths in 2022. Lung cancer represents approximately one in 8 cancers diagnosed (12.40%) and one in 5 deaths (18.70%). Lung cancer is the leading cause of cancer morbidity and mortality in men and second only to breast cancer in women. In total, the incidence and mortality rate of lung cancer in men is approximately 2 times higher than in women. In North Africa and Eastern Europe, this rate is reported to be four to five times higher. Turkey is the country with the highest incidence of lung cancer in men. Lung cancer has many genetic and non-genetic risk factors. It is known that polymorphic changes observed in genes associated with enzymes involved in DNA repair mechanisms constitute an important stepping stone for lung cancer risk. Disruptions in genes involved in the production of interleukins that stimulate immune system cells and many other genes such as cyclooxygenase enzymes involved in inflammation, p53 gene which is a tumor suppressor gene, KRAS and EGFR genes which are oncogenes, pose a threat of lung cancer. In addition, tobacco and tobacco products are among the most important risk factors for lung cancer. Polycyclic aromatic hydrocarbons and N- nitrozoamine in tobacco products have a carcinogenic effect. Studies have shown that the three-dimensional structure of deoxyribonucleic acid (DNA), which has become a molecular target with the elucidation of its double-stranded structure, can be altered without changing the nucleotide sequence, thus changing its function. DNA is also gaining recognition in the search for anticancer drug molecules. The aim is for molecules to interact with DNA through various chemical bonds (hydrophobic, ionic, hydrogen bonding and Van der Waals) in the form of electrosatellite, intercalation or groove binding. And also to treat a variety of conditions, including cancer, genetic disorders and infectious diseases, by directly affecting the structure of DNA, replication, transcription, transcription, repair or other fundamental cellular processes involving DNA. Although it is known that various drugs targeting DNA are used in cancer treatment in clinic, the search for more selective and potent drug molecules that directly affect various cellular processes by interacting with DNA, increase treatment efficacy and minimize side effects by targeting molecules or pathways necessary for cancer growth continues.

[0007] In the development of Palchaudhuri and Hergenrother anti-cancer molecules, it is important to investigate drug-protein interactions as well as their interactions with DNA. Albumin, the most abundant protein in human blood, is a good candidate anti-cancer molecule carrier. With a long half-life, albumin facilitates colloidal solubilization and transport of long-chain fatty acids, amino acids, hormones, metal ions, and various drugs in the body. Furthermore, the fact that they are taken up and metabolized at a higher rate by rapidly growing cancer cells suffering from nutrient deficiency provides a significant advantage in targeting these areas. Today, despite the use of methods such as surgery, radiotherapy, and chemotherapy in the treatment of cancer, it is known that the methods used have side effects that negatively affect human life, such as neurological, dermatological, gastrointestinal, swallowing difficulties, hematological, and cardiac problems. Therefore, alternative treatment strategies and new molecules with safer side effect profiles are needed. Photodynamic Therapy (PDT), one of these alternative treatment methods, is used in the diagnosis and treatment of many types of cancer. The basis of Photodynamic Therapy (PDT) is based on the formation of reactive oxygen types as a result of photochemical reactions of molecular oxygen in the environment as a result of exposure of a photosensitive substance to light. It has been reported that reactive types interact with biomolecules (DNA, protein, etc.) and cause cell death. Photodynamic Therapy (PDT) has the advantages of low toxic activity in the dark, low post-treatment recurrence rate and minimally invasive treatment compared to the commonly preferred treatment methods.

[0008] In Photodynamic Therapy (PDT), photosensitizing agents such as hematoporphyrin and photofrin are frequently preferred in clinic, however, these agents have a number of negative effects such as their mixed chemical structure, low tumor selectivity, and prolonged excretion from the body.

[0009] Therefore, the new photosensitizing compounds planned for synthesis are expected to possess certain properties, such as purity of chemical structure, solubility in commonly used solvents, ease of synthesis steps, low cost, high singlet oxygen yield, and strong absorption between 600-800 nm. In this context, one of the most preferred molecule groups is phthalocyanines. The metal ion in the center of the phthalocyanine ring strongly affects the chemical and physical properties of the compound. The effects of phthalocyanine compounds containing silicon in the central cavity against various types of cancer have been investigated and found to be effective in in vitro, in vivo, and clinical studies in the literature. Pyridine analogs, the substituted group preferred to use in this study, are known to have rich biological and pharmacological effects. In addition, compounds bearing substituted groups positioned in the axial position are thought to reduce aggregation and this is thought to increase the effect of Photodynamic Therapy (PDT). It also increases the water solubility of the compounds by converting the quaternary ammonium structures in the pyridine structure into pyridinium group. Water- soluble photosensitizing compounds are also preferred for ease of application in Photodynamic Therapy (PDT).

[0010] In the patent with the registration number 201814010 in the state of the art, silicon phthalocyanine compounds that can interact with DNA, show topoisomerase inhibition and have anti-cancer effect are mentioned.

[0011] In the patent with the registration number 201814012, the anti-cancer properties of water-soluble silicon phthalocyanine compounds against lung, liver, breast, and melanoma cancer types were investigated.

[0012] In the patent with registration number 201923135, water soluble cobalt (ii) phthalocyanine compounds with acetylcholinesterase inhibitory effect were investigated.

[0013] Considering the studies in the state of the art, it is important that the synthesized compounds provide the solubility in water necessary for direct injection into the body in order to be used as drugs.

[0014] Considering the above explanations and studies, silicon phthalocyanine compounds containing anti-cancer (cancer therapeutic) pyridine side group which can be used as a potential anti-tumor drug, whose spectrum of action is wider than metal-containing compounds, whose toxic effect is lower than metal-containing compounds, whose side effects are reduced, and which, thanks to their water-solubility, can be directly injected into the body, have been synthesized.

[0015] Problems to Be Solved by the Invention

[0016] The object of this study was to investigate the DNA / BSA interactions of silicon (IV) phthalocyanine (DT-2,3-SiQ compound) containing a water-soluble pyridine side group and its in vitro photodynamic therapy (PDT) activity. Lung cancer, one of the most common types of cancer, can be caused by many genetic and non-genetic causes. According to reports published by the World Health Organization in 2024, approximately 2.5 million new cases of lung cancer were diagnosed, while approximately 1.8 million people lost their lives due to this disease. The incidence and mortality of this disease is increasing rapidly worldwide.

[0017] Considering that chemotherapeutic drugs such as carboplatin, doxorubicin and cisplatin, which are most commonly used in the treatment of lung cancer in clinic, have serious side effects, developing new strategies and molecules against this disease is among the priorities of scientists. Photodynamic Therapy (PDT) has become an area of interest in recent years because it selectively destroys tumor tissue in the presence of photosensitizer, oxygen, and light and has a low side effect profile.

[0018] The combination of pyridine group and silicon phthalocyanine compounds, which are known in the literature to contain groups rich in biological activity, is thought to be very valuable in terms of being new agents with the potential to be used in Photodynamic Therapy (PDT). There are a limited number of studies in the literature investigating the Photodynamic Therapy (PDT) effect of silicon compounds against lung cancer cell lines.

[0019] Due to the high incidence and mortality rates of lung cancer, the Photodynamic Therapy (PDT) effect potentials of the inventive compounds on the A549 cell line and their relationship with apoptotic processes constitute the technical effect of the invention. The DNA interaction of the inventive compounds introduced to the literature for the first time with the A549 cell line and their in vitro photodynamic therapy (PDT) efficacy have been investigated for the first time.

[0020] Drawings

[0021] Fig. 1a: Hydrolytic and photonuclease activities of DT-2,3-SiPcQ compound (dark)

[0022] Fig. 1 b: Hydrolytic and photonuclease activities of DT-2,3-SiPcQ compound (5 min light)

[0023] Fig. 1c: Hydrolytic and photonuclease activities of DT-2,3-SiPcQ compound (15 min dark)

[0024] Fig. 1d: Hydrolytic and photonuclease activities of DT-2,3-SiPcQ compound (30 min light)

[0025] Fig. 1e: Hydrolytic and photonuclease activities of DT-2,3-SiPcQ compound (60 min dark) Detailed Description of the Invention

[0026] STEP 1 The first step of the invention is the synthesis of the compound 2,3-bis(pyridin-4- ylthio)propan-1 -ol. The structure of the synthesis is given below in Formula 2.

[0027] FORML 2 STEP 2.

[0028] In the second step of the invention, 2,3-bis(pyridin-4-ylthio)propan-1-ol compound synthesized in the first step is used to synthesize 2,3-bis(pyridin-4- ylthio)propoxy)phthalocyaninato silicon (IV) compound. This synthesis is given below in Formula s.

[0029]

[0030] STEP 3. In the third stage of the invention, the 2,3-bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) iodide compound is obtained by using the 2,3-bis(pyridin-4- ylthio)propoxy)phthalocyaninato silicon (IV) compound synthesized in the second step. The diagram is shown below in Formula 1 .

[0031]

[0032] The invention relates to a water-soluble silicon phthalocyanine compound containing 2,3-bis(pyridin-4-thio)propoxy groups and a method of synthesis of this compound. The synthesized silicon phthalocyanine compound containing 2,3-bis(pyridin-4- ylthio)propoxy groups with the molecular name 2,3-bis(pyridin-4- thio)propoxy)phthalocyaninato silicon (IV) iodide is shown in Formula 1 .

[0033] Hydrolytic and photonuclease activities of DT-2,3-SiPcQ are shown in Figure 1 ; a) dark, b) 5 min light c) 15 min light d) 30 min light e) 60 min light. (Light: white, dose: 17.5 mW / cm2). Band 1 : DNA control; bands 2-4: DNA + 1 ,3 (0.5 pM - 1 pM - 10 pM); bands 5-7: DNA + 2,3 (0.5 pM - 1 pM - 10 pM); bands 8-10: DNA + Methylene blue (0.5 pM - 1 pM - 10 pM).

[0034] In vitro studies following the synthesis of the compound shown in Formula 1 , it is shown that it damages DNA in the presence of light and has phototoxic activity in lung cancer cell lines (Fig. 1a-1e). Due to the gastrointestinal problems, hematological, neuronal, dermatological, neuronal, dermatological, and cardiac side effects observed in anti-tumor drugs in the state of the art, target-specific new water-soluble silicon phthalocyanines of Formula 1 having alternative therapeutic strategies with a safer side effect profile have been introduced. Table 1. IC50 values of DT-2,3-SiPcQ compound in A549 cell line

[0035] In the in vitro studies carried out with the compound indicated by Formula 1 in Table 1 , it was observed that the compound did not damage DNA in the dark and damaged DNA upon light application. In addition, when its effects on lung cancer cell line were examined, it was revealed that it showed strong phototoxic effect.

[0036] The method of synthesis of the compound given in Formula 1 is given below in steps:

[0037] • Adding pyridine-4-thiol and potassium ter-butylate to a single-necked flask and stirring in dry DMSO until homogeneous,

[0038] • Removing dissolved oxygen several times by adding 2,3-dibromo propanol dropwise onto the dissolved mixture with a dropping funnel,

[0039] • Stirring the reaction mixture for two days at 70 °C under nitrogen atmosphere,

[0040] • Completely evaporating the mixture cooled to room temperature,

[0041] • Dissolving of the crude product remaining after evaporation in 100 ml of chloroform and extracting three times with 100 ml of distilled water,

[0042] • Drying the post-extraction organic phase with MgSO4to evaporate the chloroform,

[0043] • Eluting the obtained crude product through the aluminum oxide-packed column into 2 fractions, Fraction 2, with 10 ml of MeOH and 10 ml of chloroform, purifying, and obtaining the compound 2,3-bis(pyridin-4-ylthio)propan-1 -oL

[0044] • Adding to a one-neck flask 165 mg (0.267 mmol) silicon phthalocyanine di chloride and 150 mg (0.534 mmol) 2,3-bis(pyridin-4-ylthio)propan-1 -ol and dissolving in 20 ml toluene,

[0045] • Stirring for 10 minutes in a nitrogen atmosphere at room temperature,

[0046] • Quickly adding a spatula tip of NaH to the mixture obtained and removing dissolved oxygen under nitrogen atmosphere several times,

[0047] • Refluxing at 110 °C for 24 hours under nitrogen atmosphere,

[0048] • Completely evaporating the solvent of the mixture cooled to room temperature, • Eluting the crude product obtained from the column with dichloromethane and ethanol using neutral silica filler and purifying to obtain 2,3-bis(pyridin-4- ylthio)propoxy)phthalocyaninato silicon (IV) compound,

[0049] • Adding 2,3-bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) compound in a 50 ml flask and dissolving with 3 ml chloroform,

[0050] • Adding 2.5 ml of methyl iodide CH3I and stirring for 3 days at room temperature, covered with aluminum foil and sealed.

[0051] • Filtering the precipitated product in the medium, washing with chloroform, hexane, and drying in a vacuum oven to obtain the water soluble 2,3- bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) iodide (1 a) compound.

[0052] Industrial Application of the Invention

[0053] According to WHO reports, cancer is among the leading causes of death worldwide. Today, the methods used in lung cancer treatment are known to have serious side effects. Therefore, alternative treatment strategies and new molecules with safer side effect profiles are needed. The successful completion of the proposed project will lead to the emergence of new agents with the potential to be used in Photodynamic Therapy (PDT), an alternative method in the treatment of lung cancer, which has a high incidence and is very difficult to treat. Since the compounds will be used for the first time, it is thought to have patent potential if the results are effective.

Claims

CLAIMS1 . A water-soluble compound of formula 1 , characterized in that,FORMOL it is a silicon phthalocyanine compound containing 2,3-bis(pyridin-4- ylthio)propoxy) groups and having the formula 2,3-bis(pyridin-4- ylthio)propoxy)phthalocyaninato silicon (IV) iodide (1a).

2. The water-soluble Formula 1 according to claim 1 , characterized in that it has phototoxic activity in human lung (A549) cancer cell lines.

3. A method of synthesis of the water-soluble compound according to claim 1 , characterized in that it comprises the process steps of:• adding pyridine-4-thiol and potassium ter-butylate to a single-necked flask and stirring in dry DMSO until homogeneous,• removing dissolved oxygen several times by adding 2,3-dibromo propanol dropwise onto the dissolved mixture with a dropping funnel,• stirring the reaction mixture for two days at 70 °C under nitrogen atmosphere,• completely evaporating the mixture cooled to room temperature,• dissolving of the crude product remaining after evaporation in 100 ml of chloroform and extracting three times with 100 ml of distilled water,• drying the post-extraction organic phase with MgSC to evaporate the chloroform,• eluting the obtained crude product through the aluminum oxide-packed column into 2 fractions, Fraction 2, with 10 ml of MeOH and 10 ml of chloroform, purifying, and obtaining the compound 2,3-bis(pyridin-4- ylthio)propan-1 -ol,• adding to a one-neck flask 165 mg (0.267 mmol) silicon phthalocyanine di chloride and 150 mg (0.534 mmol) 2,3-bis(pyridin-4-ylthio)propan-1-ol and dissolving in 20 ml toluene,• stirring for 10 minutes in a nitrogen atmosphere at room temperature,• quickly adding a spatula tip of NaH to the mixture obtained and removing dissolved oxygen under nitrogen atmosphere several times,• refluxing at 110 °C for 24 hours under nitrogen atmosphere,• completely evaporating the solvent of the mixture cooled to room temperature,• eluting the crude product obtained from the column with dichloromethane and ethanol using neutral silica filler and purifying to obtain 2,3-bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) compound,• adding 2,3-bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) compound in a 50 ml flask and dissolving with 3 ml chloroform,• adding 2.5 ml of methyl iodide CH3I and stirring for 3 days at room temperature, covered with aluminum foil and sealed,• filtering the precipitated product in the medium, washing with chloroform, hexane, and drying in a vacuum oven to obtain the water soluble 2,3- bis(pyridin-4-ylthio)propoxy)phthalocyaninato silicon (IV) iodide (1 a) compound.

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