ANTICANCER COMPOUND

Abstract

The invention relates to the synthesis of organic compounds with high anticancer activity; these compounds are intended for use in various applications within the pharmaceutical industry, medicine and healthcare, as well as the food and chemical sectors. The chemical properties of the organic compounds developed within the scope of this invention are geared towards providing high efficacy and stability in different industrial and biological applications.

Description

1 TARIFF ANTICANCER COMPOUND TECHNICAL FIELD The invention relates to the synthesis of organic compounds with high anticancer activity; the wording is 5 The compounds in question are used in the pharmaceutical industry, pharmacy, medicine and health fields, as well as in food and chemistry. It belongs to a structure intended for use in various sectors. Within the scope of the invention... The chemical properties of the developed organic compounds are suitable for various industrial and biological applications. It is designed to provide high efficiency and stability in applications. PREVIOUS TECHNIQUE Chalcones are valuable members of the flavonoid family and are important components of natural products. They are involved and exhibit a wide range of biological activities, optical properties, and applications in pharmaceuticals, agriculture, They attract attention with their potential for use in industrial fields such as food and perfumery. They are also intermediates in the synthesis of many biologically important compounds. It is used as a suitable precursor, especially in the synthesis of 5, 6 and 7-membered heterocyclic compounds. It consists of materials. Halogenated organic compounds are used in various industries, including pesticide production. It plays an important role in applications. Halogen-containing chalcone derivatives, protein kinase 20 It exhibits an inhibitory effect on the progression of hyperglycemia and obesity through its mechanism, Therefore, most drugs and drug candidate compounds consist of halogen-containing structures. Chalcone derivative compounds have antibacterial, antifungal, anticancer, antidiabetic, and antilumbanic properties. a wide range of biological activities including antioxidant, antimicrobial, antitumor and antituberculosis It is known to exhibit a wide range of features. 25 Heterocyclic compounds containing electron-rich nitrogen atoms are used in various biological activities. It plays an important role. Current research into the chemistry of such compounds, especially Development of nitrogen atom-containing substituted molecules that enhance biological activity It is focused. Pyrazolines are five-membered 30-membered compounds containing two adjacent nitrogen atoms in the unsaturated ring. They are heterocyclic compounds and are found in the brain, stomach, liver, pancreas, cervix, lungs, breast, and colon. It shows efficacy against various cancer cell lines, therefore it is used as a cancer agent. is preferred. 2 Pyrimidines and their derivatives are the most important heterocyclic atoms containing electron-rich nitrogen atoms. It is one of the structures. Besides being a basic component of DNA and RNA, this structure also has antitumor properties. antibacterial, antifungal, anticonvulsant, anti-HIV, antileishmaniasis, antiviral, anti-inflammatory, anti-ulcer, antiallergic, diuretic, herbicidal, antihypertensive, antioxidant and MAO / KDR stand out with their kinase enzyme inhibitor properties. Therefore, pyrimidine derivatives, 5 It is also effective in the treatment of neurological disorders and metabolic disorders. It is used. In addition to the contents described above, compounds containing the BODIPY core are also used in science and... Organic boron-fluorine compounds have wide application areas in technology; dyes, fluorescent 10 chemosensor, photodynamic therapy agent, ion sensor, solar cell component, strong UV It functions as an absorber and energy transfer unit. BODIPY compounds have high energy transfer potential. light absorption and emission capabilities, sharp absorption and fluorescence peaks, low With its Stokes shift and high quantum efficiency, it is superior to other fluorescent materials. It exhibits these characteristics. Furthermore, in photodynamic therapy (PDT) applications, 15 light-sensitive electrodes are used. Their use as agents has made these compounds important in cancer treatment. All these studies aim to overcome problems such as drug resistance, selectivity, and toxicity. This clearly demonstrates the need for the discovery of new anticancer molecules. Today, research in the field of medicinal chemistry focuses on the development of new drug candidates. It focuses on innovative synthetic methods for this. In this way, the symptoms The aim is to alleviate the effects of illness, improve quality of life, and extend life expectancy. Chalcones and heterocyclic compounds derived from them exhibit different reactivity, geometry, and It differs from classic chemotherapeutic drugs thanks to its kinetic properties. In this respect, 25 a vast chemical potential that remains untapped in modern drug design and development It offers, in particular, chalcone-pyrazoleline, chalcone-pyrimidine and chalcone-boron complexes, which are used for cancer. They are considered to be promising new groups of compounds in treatment. Drugs used in cancer treatment are generally low molecular weight and cytotoxic. Therefore, in addition to their effects, they can cause bone marrow suppression, gastrointestinal lesions, and hair loss. It can cause serious side effects such as hair loss, nausea, and the development of drug resistance. Therefore, the design of new anticancer agents that exhibit selective action and reduce the development of resistance. This is of great importance. In this process, molecular modeling methods are used in drug development. 3 It stands out as an important tool in their studies. A precursor with known biological activity. Instead of synthesizing numerous derivatives of molecules, through computer-based modeling Identifying derivatives with the targeted properties saves time and resources. This method provides information on the bond lengths, bond angles, torsional angles, and energy of molecules. levels, dipole moment, atomic charges, molecular orbitals, and electrostatic potential 5 Features such as MEPs (Meaning of Expansion) maps can be calculated. Furthermore, HOMO- can be determined using this data. LUMO energy difference, molecular hardness (η), electronegativity (χ), and MEP surface map are among the parameters considered. Physical parameters are also obtained. In conclusion, the current technique could lead to a new generation of anticancer drugs that can be used in cancer treatment. In the design of compounds, chalcone-based structures, pyrazolin, pyrimidine and BODIPY derivatives Their combined use offers a strong advantage in terms of both biological activity and chemical diversity. This reveals that it offers potential. THE PURPOSE OF THE INVENTION 15 The primary aim of the invention is to synthesize new chalcones that have not been previously synthesized or reported in the literature. High levels of chalcone-boron complex, pyrazolin and pyrimidine derivative compounds have been developed. The goal is to obtain novel organic structures with anticancer activity. The chalcone compounds numbered 1-8, described in the Detailed Description of the Invention section, and 9-15 20 Acetylated chalcone derivative compounds numbered 16–21, chalcone-boron complexes numbered 22–30 Pyrazoline derivatives numbered 31–37, pyrimidine compounds numbered 38–41 Pyrimidine derivatives containing an acetamide group have been searched in the SciFinder and Reaxys databases. It has been determined that these compounds have not been synthesized or studied before. In this respect... The invention is based on the first synthesis and characterization of original compound groups. 25 The synthesized compounds are similar to 5-fluorouracil, which is used as an anticancer drug on the market. When compared, they were observed to exhibit similar or higher anticancer activity. This result demonstrates the potential of these compounds to be used as anticancer agents, and They demonstrate that they have the potential to be alternatives to existing chemotherapeutic agents. 30 The resulting compound derivatives with high anticancer activity are used as drug precursors or drug active ingredients. Thanks to its production in our country, our dependence on the import of pharmaceutical raw materials has decreased. This will decrease, therefore it will be possible to significantly reduce the current account deficit. This situation is the same. 4 at the same time, it will support sustainability in domestic pharmaceutical production and access to raw materials. It will make it easier. The synthesis methods developed within the scope of the invention and the compounds obtained save time and cost. In this respect, it directly benefits the end user. 5 imported pharmaceutical components. By eliminating the resulting bureaucratic procedures and additional costs, both the production process and This results in an economic advantage in the final product. The invention relates to the synthesis of organic compounds with high anticancer activity; The compounds in question are used in the pharmaceutical industry, pharmacy, medicine and health fields, as well as in food and chemistry. 10 It belongs to a structure intended for use in various sectors. Within the scope of the invention... The chemical properties of the developed organic compounds are suitable for various industrial and biological applications. It is designed to provide high efficiency and stability in applications. The inventions are organically based and possess biological activity, and therefore have applications in medicine and pharmacy. These are compounds that have the potential to be used in the discovery of new drugs in various fields. In this context, It is a chemical product that potentially possesses anticancer and antiproliferative properties, etc. Our inventions include chalcone (1-8 / 9-15) compounds, chalcone-boron complexes (16-21), pyrazolin (22-30), pyrimidine (31-41)) derivative compounds contain different skeletal structures and chemically derived, they exhibit very strong anticancer activity. These properties are found in cells 20 Anticancer effects on proliferation according to NCI-60 screening methodology It was evaluated and the MTT test was used for this purpose. Accepted for evaluation. values ​​with a GI50 value less than 3.0 µg / mL and a TGI value less than 100 µg / mL Compounds with anticancer effects have been identified. This value is for 5-fluorouracil, which is a positive control. (5FU) is determined according to the anticancer drug. Also, strong anticancer properties Topoisomerase I 25 With Inhibition Test, Cell Migration Test, DNA Banding Test and Cytotoxicity Test It has been proven. Its in-vitro biological properties have been studied, and it shows high anticancer activity. These discoveries have been supported by molecular docking methods, which are in silico approaches. Epidermal Growth Factor Receptor in silico medium via molecular docking method. Individual interaction with (EGFR) and the results of this interaction binding parameter 30 The values ​​have been determined. LIST OF FIGURES Figure 8. Beas2B in normal lung cell line 1 (column 3), 9 (column 4), 16 (column 5) column) and 19 (column 6); 1 (column 7), 9 (column 8) in the A172 brain cancer cell line. column), 16 (column 9) and 19 (column 10); 1 (column 11) in C6 brain cancer cell line column), 9 (column 12), 16 (column 13) and 19 (column 14) and B35 brain cancer cell 5 lines 1 (column 15), 9 (column 16), 16 (column 17) and 19 (column 18) Demonstration of DNA degradation caused by molecules in gel electrophoresis. Beas2B control (column 1), A172 control (column 2), C6 control (column 19) and B35 control (column 20). Figure 9. Beas2B normal lung cell line: 12 (row 1), 26 (row 2); C6 brain cell line. cancer cell line 12 (row 3), 26 (row 4); A172 brain cancer cell line 12 (row 7), 26 (row 8) and B35 brain cancer cell line 12 (row 9), 26 (row 10 DNA degradation caused by molecules (line number) in gel electrophoresis medium Showing. Beas2B check (line 5), A172 check (line 6), C6 check (line 11). line) and B35 control (line 12). Figure 10. 33 (column 1), 34 (column 2), 35 (column 3), 9 (column 4), 10 (column 5) column), 11 (column 6), 22 (column 7), 23 (column 8), 24 (column 9), 1 (column 10) column), 2 (column 11), 3 (column 12), 16 (column 13), 17 (column 14) and 18 (columns 15 and 20 (column no. 1) Effect of test substances and control (DMSO) (K) on topoisomerase I. Unit human recombinant topoisomerase I and superhelical pHOT1 DNA test substances TGI The samples were incubated at 37°C for 30 minutes at their respective concentrations. After incubation, the samples were... Agarose was loaded onto a gel, electrophoresis was performed, and the gel was photographed using an imaging device. Figure 11. Test items 12 (1), 26 (2), 27 (3), 28 (4), 19 (5), 20 (6), 21 (7), 37 (8) and 38 (9). and the effect of Control (DMSO) (K) on topoisomerase I. One unit of human recombinant Topoisomerase I and superhelical pHOT1 DNA were tested with the substances at TGI concentrations. The samples were incubated at 37°C for 30 minutes. After incubation, the samples were loaded onto an agarose gel. Electrophoresis was performed and photographed using a gel imaging device. 30 Figure 12. 25 µM DNA of 34, 35, 36, 9, 10 and 11 in absence and presence [6.25 μM (8), 12.5 μM (7), 25 μM (6), 50 μM (5), 100 μM (4), 200 μM (3), 400 μM (2) and 800 μM (1)] UV–Vis absorption spectrum shown (A). Plot graph indicating binding constant [DNA] 6 & [DNA] / εa – εf (B). 25 µM of 34, 35, 36, 9, 10 and 11 in the absence and presence of BSA [6.25 μM (8), 12.5 μM (7), 25 μM (6) 50 μM (5), 100 μM (4), 200 μM (3), 400 μM (2) and 800 μM (1)] UV–Vis absorption spectrum (C). Plot graph indicating the binding constant. [BSA] & [BSA] / εa – εf (D). Figure 13. 25 µM 27 and 28 in the absence and presence of DNA [6.25 μM (8), 12.5 μM (7), 25 UV–Vis Absorption spectrum (A). Plot graph indicating binding constant [DNA] & [DNA] / εa – εf (B). 25 µM 27 and 28 in the absence and presence of BSA [6.25 μM (8), 12.5 μM (7), 25 μM (6) 50 μM (5), 100 μM (4), 200 μM (3), 400 μM (2) and 800 μM (1)] UV–Vis 10 Absorption spectrum (C). Plot graph indicating the binding constant [BSA] & [BSA] / εa – εf (D). DETAILED DESCRIPTION OF THE INVENTION The invention is organic in origin and possesses biological activity, therefore it has applications in the fields of medicine and pharmacy. It relates to the development of compounds with the potential to be used in new drug discovery. This 15 The chemical products obtained within this scope have potential anticancer and antiproliferative properties. These are new generation organic compounds that carry this characteristic. The compounds covered by the invention include various derivatives with different chemical skeletal structures: Compounds 1–8 are chalcone derivatives, compounds 9–15 are acetylated chalcone derivatives, compounds 16–20 Compounds numbered 21 are chalcone-boron complexes, compounds numbered 22–30 are pyrazolin derivatives, Compounds 31–37 are pyrimidine compounds and compounds 38–41 are acetamide group compounds. These are pyrimidine derivatives containing different chemical skeletal structures. Thanks to this, it exhibits a high level of anticancer activity. Synthesis of chalcone (1-8 / 9-15) derivative compounds The chalcone derivative compounds synthesized within the scope of this invention are: It has the compound structure (2E)-1-(X-Aminophenyl)-3-(Y)prop-2-en-1-one, where X is 2, 3 and one of the 4 values, Y; “2,4-difluorophenyl”, “3,4-difluorophenyl”, “2-chloro-5-fluorophenyl” and “2- It can be one of the values ​​of “chloro-4-chlorophenyl”. According to this structure, 30 within the scope of the invention. The synthesized chalcone derivative compounds are as follows: • (2E)-1-(2-Aminophenyl)-3-(2,4-difluorophenyl)prop-2-en-1-one (1) • (2E)-1-(3-Aminophenyl)-3-(2,4-difluorophenyl)prop-2-en-1-one (2) • (2E)-1-(2-Aminophenyl)-3-(3,4-difluorophenyl)prop-2-en-1-one (3) 7 • (2E)-1-(2-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one (4) • (2E)-1-(4-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one (5) • (2E)-1-(2-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (6) • (2E)-1-(3-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (7) • (2E)-1-(4-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (8) 5 Synthesis of compounds 1-8: Chalcone compounds were synthesized using a conventional method called Claisen-Schmidt. It was synthesized according to the method. Water-ethyl alcohol (30 mL) solvent placed in a beaker. NaOH (5-25 mmol) is added to the mixture and it is cooked at room temperature using a magnetic stirrer. It was dissolved by stirring. 2' / 4'-10 mL of ethyl alcohol was added to the prepared NaOH solution. aminoacetophenone (1-10 mmol) is added separately until completely dissolved. It is mixed. 2,4 / 3,4 / 3,5-difluorobenzaldehyde (1-10 mL) in ethyl alcohol is added to the solution. The solution was reacted separately by adding it dropwise (mmol) over 30 minutes. The reactions were continued for 12 hours at room temperature, and the reaction progress was monitored using ICT. This was done. The water content in the mixture was increased to ensure complete sedimentation. The precipitated solid was 15 The mixture was left in the refrigerator overnight to allow the substance to mature. Then it precipitated. The sample was filtered from the crucible and washed with cold pure water. The resulting yellow compounds... It was dried in a lyophilizer. A certain amount of solid was taken, dissolved in chloroform, and LTD (fine) The purity of the substance obtained by (layer chromatography) was checked. The structure of the compound... spectroscopic methods (1H-NMR, 13C-APT, 1H-1HCOSY, LC-MS and FT-IR 20 This has been clarified. The synthesis equation for the compound is given in Figure 1. Synthesis of compounds 2 and 7: Acetylated chalcone derivative compound (10 mmol), 50 mL The mixture is placed in a round-bottomed flask and 20 mL of ethanol is added. The resulting mixture... 10 mL of 6 N HCl solution was added and it was stored at 100 ºC under reflux for 16 hours. It was mixed. The progress of the reaction was monitored by ICT and terminated. Cooled 25 Ethanol was removed from the reaction mixture using an evaporator. Flask contents: 100 The mixture is placed in a mL beaker and 5-50 mL of pure water is added. It is then placed on a magnetic stirrer. The acid formed in the environment is neutralized into the reaction mixture, and the targeted To cause the compound to precipitate, 5 N NaOH solution is added dropwise to the reaction material. The addition of NaOH solution was monitored with a pH meter until pH=7, then 30... The process has been completed. At this stage, the formation of a yellow precipitate in the environment has been observed. The yellow precipitate... It was left in the refrigerator overnight to mature. Once matured, the solid was washed with pure water. It was filtered from the crucible and dried in a lyophilizer. The purity of the compound was checked by ITC. 8 The structure of the compound was determined using spectroscopic methods (1H-NMR, 13C-APT, LC-MS and FT-). (IR) has been illuminated. The synthesis scheme of the compound is given in Figure 2. Figure 1. General synthesis scheme of halosubstituted aminochalcone compounds. Figure 2. Synthesis scheme of compounds 2 and 7. (2E)-1-(2-Aminophenyl)-3-(2,4-difluorophenyl) prop-2-en-1-one (1) 9 (2E)-1-(3-Aminophenyl)-3-(2,4-difluorophenyl)prop-2-en-1-one (2) (2E)-1-(2-Aminophenyl)-3-(3,4-difluorophenyl) prop-2-en-1-one (3) 5 (2E)-1-(2-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one (4) (2E)-1-(4-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one (5) (2E)-1-(2-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (6) (2E)-1-(3-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (7) (2E)-1-(4-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one (8) Acetylated chalcone derivative compounds synthesized within the scope of the invention: 5 It has the compound structure N-{X-[(2E)-3-(Y)prop-2-enoyl]phenyl}acetamide. Here X; 2, 3 and one of the 4 values, Y; one of the values ​​“3,Y1-Difluorophenyl” and “2-Chloro-Y2-fluorophenyl”; Y1 can be either 4 or 5, and Y2 can be either 4, 5, or 6. This structure... According to the invention, the acetyl chalcone derivative compounds synthesized within the scope of the invention are as follows: • N-{4-[(2E)-3-(2,4-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (9) 10 • N-{4-[(2E)-3-(3,4-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (10) • N-{2-[(2E)-3-(3,5-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (11) • N-{2-[(2E)-3-(2-Chloro-6-fluorophenyl)prop-2-enoyl]phenyl}acetamide (12) • N-{2-[(2E)-3-(2-Chloro-5-fluorophenyl)prop-2-enoyl]phenyl}acetamide (13) • N-{3-[(2E)-3-(2-Chloro-4-fluorophenyl)prop-2-enoyl]phenyl}acetamide (14) 15 • N-{4-[(2E)-3-(2-Chloro-4-fluorophenyl)prop-2-enoyl]phenyl}acetamide (15) Synthesis of acetylated chalcone derivative compounds numbered 9-15: Acetylated chalcone derivative compounds For synthesis, 2' / 3' / 4'-aminoacetophenols are first reacted separately with acetic anhydride. inserted, N-(2-acetylphenyl)acetamide, N-(3-acetylphenyl)acetamide and N-(4-acetylphenyl)acetamide 20 The compounds were obtained via nucleophilic acyl substitution reactions. The compound... The synthesis equation is given in Figure 3. For the synthesis of acetylated chalcone derivative compounds numbered 9-15; Claisen-Schmidt condensation. The method was used. 25 ml of water-ethyl alcohol (30 ml) solvent mixture was placed in a beaker. NaOH (25 mmol) is added and stirred at room temperature using a magnetic stirrer. It has been dissolved. The prepared NaOH solution was mixed with ethyl alcohol (10 mL) containing N-(2' / 3' / 4'- Acetylphenyl acetamide (10 mmol) was added and stirred until completely dissolved. Regarding the solution; for compounds 9-11, 2-chloro-6-fluorobenzaldehyde, 2- in ethyl alcohol (10 mL). chloro-5-fluorobenzaldehyde, 2-chloro-4-fluorobenzaldehyde (10 mmol); 2.4 / -30 for compounds 12-15 3,4 / 3,5-difluorobenzaldehyde (10 mmol) was added dropwise over 30 minutes, separately. 11 The reactions were initiated. The reactions were continued for 12 hours at room temperature, and the reaction... The progress was monitored with a sedimentation test. The sedimentation was completed by increasing the water content in the mixture. This has been ensured. To allow the precipitated solid to mature, the mixture will be left in the refrigerator overnight. It was left to stand. Then the settled portions were filtered from the crucible and washed with cold, pure water. The resulting compounds were dried in a lyophilizer. A quantity of the solid was taken and mixed with chloroform. The substance was dissolved and purity control was performed using ICT. The structure of the compound... The compound was elucidated using spectroscopic methods (1H-NMR, 13C-APT, LC-MS, and FT-IR). The synthesis equation is given in Figure 4. Figure 3. General synthesis equation for N-(2 / 3 / 4-acetylphenyl)acetamide derivative compounds. Figure 4. General synthesis scheme of halosubstituted N-acetyl chalcone derivative compounds. N-{4-[(2E)-3-(2,4-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (9) 12 N-{4-[(2E)-3-(3,4-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (10) N-{2-[(2E)-3-(3,5-Difluorophenyl)prop-2-enoyl]phenyl}acetamide (11) 5 N-{2-[(2E)-3-(2-Chloro-6-fluorophenyl)prop-2-enoyl]phenyl}acetamide (12) N-{2-[(2E)-3-(2-Chloro-5-fluorophenyl)prop-2-enoyl]phenyl}acetamide (13) N-{3-[(2E)-3-(2-Chloro-4-fluorophenyl)prop-2-enoyl]phenyl}acetamide (14) 15 13 N-{4-[(2E)-3-(2-Chloro-4-fluorophenyl)prop-2-enoyl]phenyl}acetamide (15) Chalcone-boron difluoride complexes synthesized within the scope of this invention: 5 It has the compound structure (2E)-1-(2-Aminophenyl)-3-(X)prop-2-en-1-on-boron difluoride. Here, X stands for “3,4-difluorophenyl”, “3,5-difluorophenyl”, “2-chloro-6-fluorophenyl”, “2-chloro-5-fluorophenyl” and may be one of the values ​​“2-chloro-4-fluorophenyl”. According to this structure, the invention falls within the scope of this. The synthesized chalcone-boron difluoride compounds are as follows: • (2E)-1-(2-Aminophenyl)-3-(2,4-difluorophenyl)prop-2-en-1-one-borondifluoride (16) 10 • (2E)-1-(2-Aminophenyl)-3-(3,4-difluorophenyl)prop-2-en-1-one-borondifluoride (17) • (2E)-1-(2-Aminophenyl)-3-(3,5-difluorophenyl)prop-2-en-1-one-borondifluoride (18) • (2E)-1-(2-Aminophenyl)-3-(2-chloro-6-fluorophenyl)prop-2-en-1-one-borondifluoride (19) • (2E)-1-(2-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one-borondifluoride (20) • (2E)-1-(2-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one-borondifluoride (21) 15 Synthesis of Chalcone-Bordifluoride Complexes (16-21) Chalcone-boron difluoride complexes (16-21) boron trifluoride diethyl ether as Lewis acid the complex through acid-base reactions in which chalcone is also used as a Lewis base It was synthesized. Chalcone compounds (1-10 mmol) were placed in a 50 mL round-bottomed flask and 20 They were dissolved separately in 1-10 mL of dry dichloromethane. Then 1-10 mL of triethylamine was added to them. It was added and mixed under reflux for 20 minutes. Dropping into the reaction medium. The BF3.Et2O complex is added dropwise to the funnel (1-10 mL) and the reaction is continued for 4 hours. The reaction was continued. Precipitation was observed inside the flask during the reaction. The precipitate obtained after evaporating the solvent content is filtered and washed with cold dichloromethane for 25 minutes. They were then dried at room temperature. The resulting compounds were purified using ICT and Its structure was elucidated using spectroscopic methods (1H-NMR, 13C-APT, LC-MS, and FT-IR). The synthesis equation for the compound is given in Figure 9. 14 Figure 5. Synthesis scheme of chalcone-bordifluoride complexes. (2E)-1-(2-Aminophenyl)-3-(2,4-difluorophenyl)prop-2-en-1-one-borondifluoride (16) 5 (2E)-1-(2-Aminophenyl)-3-(3,4-difluorophenyl)prop-2-en-1-one-borondifluoride (17) 15 (2E)-1-(2-Aminophenyl)-3-(3,5-difluorophenyl)prop-2-en-1-one-borondifluoride (18) (2E)-1-(2-Aminophenyl)-3-(2-chloro-6-fluorophenyl)prop-2-en-1-one-borondifluoride (19) (2E)-1-(2-Aminophenyl)-3-(2-chloro-5-fluorophenyl)prop-2-en-1-one-borondifluoride (20) (2E)-1-(2-Aminophenyl)-3-(2-chloro-4-fluorophenyl)prop-2-en-1-one-borondifluoride (21) Pyralozine derivative compounds synthesized within the scope of this invention: It has the compound structure {X-[5-(Y)-Z-1H-pyrazole-3-yl]phenyl}amine, where X represents 2, 3, and 4. One of the values ​​is Y; “2,4-Difluorophenyl”, “3,5-Difluorophenyl”, “2-Chloro-6-fluorophenyl”, “2- One of the values ​​"Chloro-5-fluorophenyl" and "2-Chloro-4-fluorophenyl" is Z; the value "4,5-dihydro" and It can be one of the "empty" values. According to this structure, pyralosin 15 synthesized within the scope of the invention Derivative compounds are as follows: • {2-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (22) • {3-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (23) • {4-[5-(2,4-Difluorophenyl)-1H-pyrazol-3-yl]phenyl}amine (24) • {2-[5-(3,5-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (25) 20 • {2-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (26) 16 • {3-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (27) • {4-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (28) • {2-[5-(2-Chloro-5-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (29) • {2-[5-(2-Chloro-4-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (30) Synthesis of pyrazolin (22-30) derivative compounds Synthesis of pyrazole (22-30) derivative compounds by acid-catalyzed hydrazine in chalcones This was achieved by the Michael addition reaction of monohydrate. Chalcone compounds (5 The solution was taken separately into a 50 mL round-bottomed flask (mmol) and 20 mL of dry ethanol was added to it. hydrazine monohydrate (10 mmol), 2 mL glacial acetic acid 10 ml were added to the resulting solution. The reaction mixtures were mixed at 80 °C for 6 hours under reflux. The reaction progress was monitored by TCT, terminated, and the cooled reaction contents were salted. It was poured over cold water containing [the substance] and precipitated. The resulting white solid substances came into contact with air. It was washed with pure water in a way that would not cause any damage, filtered from the crucible, and dried in a lyophilizer. The purity of the compounds was determined by ICT and their structure was analyzed using spectroscopic methods (1H-NMR, 15 13C-APT, LC-MS and FT-IR) elucidation was performed. The synthesis scheme of the compound is given in Figure 6. Figure 6. General synthesis scheme of pyrazolein compounds. {2-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (22) 17 {3-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (23) {4-[5-(2,4-Difluorophenyl)-1H-pyrazol-3-yl]phenyl}amine (24) 5 {2-[5-(3,5-Difluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (25) {2-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (26) {3-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (27) 18 {4-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (28) {2-[5-(2-Chloro-5-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (29) {2-[5-(2-Chloro-4-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl]phenyl}amine (30) Pyrimidine derivative compounds synthesized within the scope of the invention: 10 In one of the inventions, the compound structure is 4-(X-Aminophenyl)-6-(Y)pyrimidine-2-amine. It has. Here, X is one of the values ​​2, 3, and 4, and Y is “2,4-Difluorophenyl” or “3,4-Difluorophenyl”. “3,5-Difluorophenyl”, “2-chloro-4-fluorophenyl”, “2-chloro-5-fluorophenyl” and “2-chloro-6-fluorophenyl” It can be one of the values. In another structure of the invention, N-(3-(X-Amino-6- (Y)pyrimidine-4-yl)phenyl)acetamide has a compound structure. Here, X is 15 from the values ​​2, 3 and 4. One of them, Y; one of the values ​​“2,4-Difluorophenyl”, “3,5-Difluorophenyl” and “2-chloro-5-fluorophenyl”. This is possible. According to this structure, pyrimidine derivative compounds synthesized within the scope of the invention. It is as follows: • 4-(4-Aminophenyl)-6-(2,4-difluorophenyl)pyrimidine-2-amine (31) • 4-(3-Aminophenyl)-6-(3,4-difluorophenyl)pyrimidine-2-amine (32) 20 • 4-(2-Aminophenyl)-6-(3,5-difluorophenyl)pyrimidine-2-amine (33) • N-(3-(3-Amino-6-(2,4-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (34) • N-(3-(3-Amino-6-(3,5-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (35) • N-(3-(4-Amino-6-(3,5-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (36) • 4-(3-Aminophenyl)-6-(2-chloro-6-fluorophenyl)pyrimidine-2-amine (37) 25 • 4-(4-Aminophenyl)-6-(2-chloro-6-fluorophenyl)pyrimidine-2-amine (38) • 4-(2-Aminophenyl)-6-(2-chloro-5-fluorophenyl)pyrimidine-2-amine (39) • 4-(4-Aminophenyl)-6-(2-chloro-4-fluorophenyl)pyrimidine-2-amine (40) 19 • N-(3-(2-Amino-6-(2-chloro-5-fluorophenyl)pyrimidin-4-yl)phenyl)acetamide (41) Synthesis of pyrimidine (31-41) derivative compounds Synthesis of pyrimidine (31-41) derivative compounds, conjugation of guanidine hydrochloride (1,4- This was performed with Michael's addition. Chalcone compounds (5 mmol) were placed in 50 mL round bottles. Each sample was placed separately into a flask and dissolved in 10 mL of DMSO. Guanidine hydrochloride (5 mmol) and solid NaOH (5 mmol) were added. Reaction components 12 The reaction was stirred under a cooling system for an hour. The reaction was terminated by monitoring with ITK. Pure water was added to the mixtures. The resulting solid particles were washed away with pure water, filtered, and It was dried in a lyophilizer. The purity of the obtained solids was checked with ICT and the impurity level was 10 Purified by solid column chromatography. The structure of the obtained compounds is as follows: illuminated by spectroscopic methods (1H-NMR, 13C-APT, LC-MS and FT-IR) This has been confirmed. The general synthesis scheme of the compounds is given in Figure 7. Figure 7. General synthesis scheme of pyrimidine compounds. 4-(4-Aminophenyl)-6-(2,4-difluorophenyl) pyrimidine-2-amine (31) 20 4-(3-Aminophenyl)-6-(3,4-difluorophenyl) pyrimidine-2-amine (32) 4-(2-Aminophenyl)-6-(3,5-difluorophenyl)pyrimidine-2-amine (33) N-(3-(3-Amino-6-(2,4-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (34) N-(3-(3-Amino-6-(3,5-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (35) 21 N-(3-(4-Amino-6-(3,5-difluorophenyl)pyrimidine-4-yl)phenyl)acetamide (36) 4-(3-Aminophenyl)-6-(2-chloro-6-fluorophenyl)pyrimidine-2-amine (37) 5 4-(4-Aminophenyl)-6-(2-chloro-6-fluorophenyl)pyrimidine-2-amine (38) 4-(2-Aminophenyl)-6-(2-chloro-5-fluorophenyl)pyrimidine-2-amine (39) 4-(4-Aminophenyl)-6-(2-chloro-4-fluorophenyl)pyrimidine-2-amine (40) 22 N-(3-(2-Amino-6-(2-chloro-5-fluorophenyl)pyrimidin-4-yl)phenyl)acetamide (41) Performing and Evaluating Anticancer Activity Tests of Invention Compounds a. Evaluation of Anticancer Activity According to NCI60 Screening Methodology 5 The anticancer effects of the discovered compounds on cell proliferation were screened using the NCI-60 method. The MTT test was used to evaluate according to its methodology. Accepted for evaluation. The values ​​obtained were GI50 less than 3.0 µg / mL and TGI less than 100 µg / mL. Compounds with anticancer effects have been identified. This value is positive for 5-fluorouracil as a control. (5FU) was determined according to the anticancer drug. The LC50 value was 10 times less than 500 µg / mL in cancer cells. small but normal cells should be at least 5FU or larger than the control drug. It is expected. Table 1. GI50, TGI, and LC50 values ​​of chalcone compounds in cells A172, C6, and B35. Compound (µg / mL) A172 C6 B35 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 1 31.5 >500 >500 2.2 240.3 >500 1.9 11.1 >500 2 1.8 5.91 207.2 1.6 7.6 >500 1.2 2.2 15.0 3 8.1 >500 >500 1.6 8.4 >500 1.3 2.5 19.6 4 >500 >500 >500 1.8 317.6 >500 2.6 98.3 >500 391.4 >500 >500 2.0 >500 >500 3.4 >500 >500 6 59.5 >500 >500 1.4 5.9 >500 1.8 9.2 >500 7 196.5 >500 >500 1.9 32.8 >500 1.8 7.7 >500 8 >500 >500 >500 2.1 >500 >500 1.7 22.0 >500 5FU 1.4 71.4 423.8 1.5 77.2 481.6 1.4 78.3 469.1 Table 2. GI50, TGI and LC50 of chalcone compounds in A549, Calu1 and H1650 cells. values Compound (µg / mL) A549 Calu1 H1650 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 1 >500 >500 >500 2.4 >500 >500 1.0 21.7 >500 2 3.3 99.8 >500 1.4 7.6 >500 1.0 1.7 >500 3 5.4 >500 >500 1.0 >500 >500 1.2 7.6 >500 4 >500 >500 >500 1.6 >500 >500 1.6 >500 >500 >500 >500 >500 1.4 >500 >500 1.3 394.0 >500 6 26.4 >500 >500 1.2 >500 >500 1.3 >500 >500 23 7 27.1 >500 >500 1.3 313.2 >500 1.2 12.5 >500 8 >500 >500 >500 1.1 >500 >500 1.0 >500 >500 5FU 1.3 54.6 458.0 1.6 68.2 479.3 1.5 45.7 409.5 Table 3. GI50, TGI, and LC50 of chalcone compounds in Beas2B and HC normal cells. values Compound (µg / mL) Beas2B HC GI50 TGI LC50 GI50 TGI LC50 1 2.4 265.1 >500 1.3 >500 >500 2 1.8 8.8 >500 1.3 3.1 74.8 3 1.2 3.5 >500 1.2 3.2 361.2 4 1.6 6.4 >500 1.3 79.6 >500 2.5 >500 >500 1.2 >500 >500 6 1.3 4.3 >500 1.6 9.4 >500 7 1.6 19.6 >500 1.4 11.7 >500 8 1.6 10.7 >500 1.6 >500 >500 5FU 1.3 55.3 427.6 1.5 82.4 405.9 Table 4. GI50, TGI and LC50 of acetylated chalcone compounds in cells A172, C6 and B35. values Compounds (µg / mL) A172 C6 B35 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 9 3.0 >500 >500 2.5 166.0 >500 1.4 2.8 21.6 1.6 13.9 >500 4.8 >500 >500 1.7 9.7 >500 11 1.4 55.6 >500 293.5 >500 >500 1.4 18.2 >500 12 2.8 >500 >500 >500 >500 >500 1.1 2.0 16.6 13 5.3 >500 >500 >500 >500 >500 2.5 134.6 >500 14 3.0 >500 >500 4.0 >500 >500 1.3 3.1 138.5 1.1 168.4 >500 4.0 >500 >500 1.3 3.4 114.5 5FU 1.4 71.4 423.8 1.5 77.2 481.6 1.4 78.3 469.1 Table 5. Acetylated chalcone compounds in A549, Calu1 and H1650 cells GI50, TGI and LC50 values Compound (µg / mL) A549 Calu1 H1650 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 9 171.3 >500 >500 1.4 467.3 >500 1.0 23.1 >500 >500 >500 >500 1.8 143.6 >500 1.3 8.3 >500 11 >500 >500 >500 1.3 >500 >500 1.1 >500 >500 12 >500 >500 >500 1.7 >500 >500 11.8 >500 >500 13 >500 >500 >500 1.7 >500 >500 1.6 >500 >500 24 14 >500 >500 >500 1.2 >500 >500 1.0 1.9 473.5 >500 >500 >500 1.1 50.5 >500 1.1 4.3 >500 5FU 1.3 54.6 458.0 1.6 68.2 479.3 1.5 45.7 409.5 Table 6. GI50, TGI, and LC50 of acetylated chalcone compounds in Beas2B and HC cells. values Compounds (µg / mL) Beas2B HC GI50 TGI LC50 GI50 TGI LC50 9 1.0 1.4 >500 1.2 12.7 >500 1.0 1.5 >500 1.5 6.8 >500 11 1.0 1.0 >500 1.4 >500 >500 12 1.0 2.7 >500 1.0 3.5 >500 12 1.0 2.2 >500 1.3 >500 >500 14 1.0 1.3 30.4 1.4 11.5 >500 1.2 1.5 >500 1.2 35.6 >500 5FU 1.3 55.3 427.6 1.5 82.4 405.9 Table 7. Compounds of chalcone boron complexes in cells A172, C6 and B35 GI50, TGI and LC50 values Compounds (µg / mL) A172 C6 B35 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 16 12.0 >500 >500 1.2 22.6 >500 1.4 9.5 >500 17 6.3 >500 >500 1.3 8.3 >500 1.2 3.9 >500 18 6.6 >500 >500 1.3 4.0 >500 1.2 2.5 40.4 19 1.1 3.0 >500 1.2 4.9 >500 1.3 6.0 >500 2.4 >500 >500 1.1 >500 >500 1.4 >500 >500 21 1.2 22.3 >500 1.1 50.0 >500 1.0 3.8 >500 5FU 1.4 71.4 423.8 1.5 77.2 481.6 1.4 78.3 469.1 Table 8. Compounds of chalcone boron complexes in cells A549, Calu1 and H1650. GI50, TGI and LC50 values Compounds (µg / mL) A549 Calu1 H1650 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 16 >500 >500 >500 1.0 444.7 >500 1.4 7.9 >500 17 8.8 >500 >500 1.0 1.6 >500 1.4 11.2 >500 18 5.4 >500 >500 1.0 >500 >500 1.2 4.3 >500 19 29.9 >500 >500 1.2 5.2 >500 1.0 1.4 117.8 >500 >500 >500 1.1 492.3 >500 1.1 >500 >500 21 >500 >500 >500 1.8 >500 >500 1.2 >500 >500 5FU 1.3 54.6 458.0 1.6 68.2 479.3 1.5 45.7 409.5 Table 9. GI50, TGI of chalcone boron complex compounds in Beas2B and HC cells. and LC50 values Compounds (µg / mL) Beas2B HC GI50 TGI LC50 GI50 TGI LC50 16 1.4 79.5 >500 1.1 19.5 >500 17 1.5 16.9 >500 1.1 5.4 >500 18 1.6 8.8 >500 1.1 2.7 >500 19 1.6 17.8 >500 1.0 2.1 >500 2.3 >500 >500 1.0 >500 >500 21 1.3 4.4 >500 1.1 4.0 >500 5FU 1.3 55.3 427.6 1.5 82.4 405.9 Table 10. GI50, TGI, and LC50 values ​​of pyrazolin compounds in cells A172, C6, and B35. Compounds (µg / mL) A172 C6 B35 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 22 4.6 >500 >500 1.6 72.7 >500 1.7 147.8 >500 23 5.2 >500 >500 1.3 30.6 >500 1.8 29.4 >500 24 6.1 >500 >500 1.6 58.3 >500 1.5 49.8 >500 2.6 >500 >500 1.6 >500 >500 1.1 >500 >500 26 2.3 >500 >500 29.6 >500 >500 5.9 >500 >500 27 3.8 >500 >500 4.4 >500 >500 2.9 >500 >500 28 4.1 >500 >500 2.0 >500 >500 1.8 >500 >500 29 1.3 12.1 >500 1.4 146.5 >500 1.3 90.6 >500 9.8 >500 >500 3.9 >500 >500 3.4 >500 >500 5FU 1.4 71.4 423.8 1.5 77.2 481.6 1.4 78.3 469.1 Table 11. GI50, TGI, and LC50 of pyrazolin compounds in A549, Calu1, and H1650 cells. values Compounds (µg / mL) A549 Calu1 H1650 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 22 >500 >500 >500 1.0 44.5 >500 1.0 3.6 >500 23 >500 >500 >500 1.8 261.8 >500 1.0 3.3 >500 24 >500 >500 >500 1.4 33.0 >500 1.0 4.1 >500 >500 >500 >500 1.2 >500 >500 1.0 >500 >500 26 >500 >500 >500 1.2 52.4 >500 1.3 >500 >500 27 >500 >500 >500 1.0 3.1 >500 1.1 2.5 >500 28 >500 >500 >500 1.3 12.8 149.2 1.1 5.7 >500 29 >500 >500 >500 1.0 9.4 >500 1.0 2.3 >500 >500 >500 >500 1.7 >500 >500 1.0 18.0 >500 5FU 1.3 54.6 458.0 1.6 68.2 479.3 1.5 45.7 409.5 Table 12. GI50, TGI, and LC50 values ​​of pyrazolin compounds in Beas2B and HC cells. Compounds (µg / mL) Beas2B HC GI50 TGI LC50 GI50 TGI LC50 22 3.5 >500 >500 1.0 >500 >500 23 5.0 >500 >500 1.0 491.2 >500 24 2.8 >500 >500 1.1 227.1 >500 >500 >500 >500 1.1 >500 >500 26 1.0 >500 >500 2.5 >500 >500 27 1.2 40.6 >500 1.6 >500 >500 28 1.1 10.5 >500 1.1 >500 >500 26 29 1.4 212.2 >500 1.0 >500 >500 2.6 >500 >500 1.1 >500 >500 5FU 1.3 55.3 427.6 1.5 82.4 405.9 Table 13. GI50, TGI, and LC50 values ​​of pyrimidine compounds in cells A172, C6, and B35. Compounds (µg / mL) A172 C6 B35 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 31 1.9 >500 >500 3.7 >500 >500 1.5 19.9 >500 32 2.6 >500 >500 14.0 >500 >500 3.1 >500 >500 33 1.6 >500 >500 1.5 71.3 >500 1.2 34.7 >500 34 1.0 >500 >500 1.9 >500 >500 1.0 >500 >500 35 1.0 18.2 >500 1.4 >500 >500 1.1 30.1 >500 36 13.3 >500 >500 1.0 52.7 >500 1.1 67.1 >500 37 6.8 >500 >500 1.2 >500 >500 2.1 >500 >500 38 13.8 >500 >500 1.2 >500 >500 1.4 >500 >500 39 1.5 >500 >500 2.5 >500 >500 1.7 >500 >500 40 1.0 12.8 >500 1.8 >500 >500 1.1 9.4 >500 41 25.9 >500 >500 2.3 >500 >500 1.1 >500 >500 5FU 1.4 71.4 423.8 1.5 77.2 481.6 1.4 78.3 469.1 Table 14. GI50, TGI, and LC50 of pyrimidine compounds in A549, Calu1, and H1650 cells. values Compounds (µg / mL) A549 Calu1 H1650 GI50 TGI LC50 GI50 TGI LC50 GI50 TGI LC50 31 >500 >500 >500 1.0 43.4 >500 1.0 212.4 >500 32 >500 >500 >500 1.3 >500 >500 1.0 >500 >500 33 >500 >500 >500 1.0 1.3 >500 1.0 >500 >500 34 >500 >500 >500 1.0 8.8 >500 1.0 5.5 >500 35 >500 >500 >500 1.0 1.3 >500 1.0 6.6 >500 36 >500 >500 >500 1.2 409.9 >500 >500 >500 >500 37 >500 >500 >500 1.0 >500 >500 1.3 >500 >500 38 >500 >500 >500 1.0 >500 >500 1.1 >500 >500 39 >500 >500 >500 1.1 >500 >500 1.0 >500 >500 40 >500 >500 >500 1.0 3.1 >500 2.1 >500 >500 41 >500 >500 >500 1.0 5.6 >500 1.0 >500 >500 5FU 1.3 54.6 458.0 1.6 68.2 479.3 1.5 45.7 409.5 27 Table 15. GI50, TGI, and LC50 values ​​of pyrimidine compounds in Beas2B and HC cells. Compounds (µg / mL) Beas2B HC GI50 TGI LC50 GI50 TGI LC50 31 1.0 1.2 >500 1.0 >500 >500 32 1.2 16.0 >500 1.0 >500 >500 33 >500 >500 >500 1.0 388.3 >500 34 1.8 13.9 >500 1.0 >500 >500 35 2.5 >500 >500 1.4 >500 >500 36 1.0 >500 >500 1.0 >500 >500 37 18 2.9 >500 >500 1.0 >500 38 19 3.2 >500 >500 1.1 >500 39 20 12.7 >500 >500 1.0 >500 40 21 1.1 >500 >500 1.0 >500 41 22 1.7 10.5 >500 1.0 >500 5FU 1.3 55.3 427.6 1.5 82.4 405.9 Evaluation of the Cytotoxic Activities of Synthesized Compounds 5. Measurement of cytoplasmic LDH enzyme provides some information about the cytotoxic activities of compounds. It provides cytoplasmic lactate dehydrogenase activity, using an LDH cytotoxicity kit. The cytotoxicity caused by the compounds synthesized within the scope of this thesis is measured using TGI. Measurements were taken using concentrations. The measured LDH leakage value was in the range of 0%-10%. Those that were present were considered non-cytotoxic. Measured LDH leakage values ​​were 11%-20%. Those within this range are moderately cytotoxic, and those above this value are highly cytotoxic. It was considered cytotoxic. These values ​​were compared with 5FU anticancer, which was used as a positive control. It was determined according to the agent. Table 16. The effects of chalcone compounds on cells at 15% TGI concentrations. Cytotoxicity values % Cytotoxicity A549 Calu1 H1650 A172 C6 B35 Beas2B HC 1 3.0 3.3 12.5 3.2 8.1 22.1 7.7 3.9 2 8.6 24.2 29.8 25.6 27.3 29.0 24.4 28.9 3 2.9 3.5 27.6 3.4 24.6 27.9 28.6 27.1 4 2.1 3.6 2.5 3.5 7.2 9.8 25.0 10.0 2.2 2.8 6.7 2.9 2.8 2.5 3.7 3.6 6 2.2 2.6 2.6 2.7 28.2 24.3 28.1 28.2 7 2.7 7.3 21.6 2.6 15.8 26.6 12.2 24.5 8 3.1 2.7 3.3 3.0 2.7 27.0 24.3 3.8 5FU 15.5 16.3 17.1 14.9 16.7 15.0 17.2 16.4 28 Table 17. Acetylated chalcone compounds in cells at TGI concentrations. % Cytotoxicity values ​​exhibited against % Cytotoxicity A549 Calu1 H1650 A172 C6 B35 Beas2B HC 9 3.4 5.4 11.7 3.3 6.1 29.8 29.1 21.2 3.4 7.7 25.1 21.2 3.4 25.6 28.7 25.9 11 3.5 2.7 2.8 12.1 3.5 14.5 29.2 3.0 12 2.9 3.4 2.9 3.8 3.8 28.1 28.1 27.5 13 3.1 3.5 2.9 3.2 3.7 6.6 29.5 2.9 14 3.1 3.1 29.0 3.2 3.8 29.3 29.8 25.4 2.8 10.1 29.0 5.9 3.5 29.4 28.5 11.0 5FU 15.5 16.3 17.1 14.9 16.7 15.0 17.2 16.4 Table 18. Effects of chalcone-boron complexes on cells at TGI concentrations. % Cytotoxicity values % Cytotoxicity A549 Calu1 H1650 A172 C6 B35 Beas2B HC 16 3.8 5.7 25.7 3.5 18.1 27.2 10.0 17.3 17 3.7 28.2 22.2 2.0 23.4 27.1 12.5 26.2 18 3.7 3.6 21.9 2.1 27.2 28.1 24.7 29.2 19 3.7 26.8 29.3 27.6 24.3 22.0 10.8 28.4 3.5 5.3 3.1 3.4 2.0 2.2 3.7 2.1 21 3.9 3.5 3.3 12.8 10.70 27.0 28.1 27.1 5FU 15.5 16.3 17.1 14.9 16.7 15.0 17.2 16.4 Table 19. 10% TGI concentrations exhibited by pyrazole compounds against cells. Cytotoxicity values % Cytotoxicity A549 Calu1 H1650 A172 C6 B35 Beas2B HC 22 2.9 10.3 27.1 2.8 10.1 9.3 3.6 3.8 23 3.0 7.5 27.0 3.0 18.1 19.8 2.8 5.7 24 3.2 10.0 24.0 2.9 10.2 17.4 2.7 5.9 3.2 3.5 2.7 2.9 3.4 3.6 2.6 3.2 26 3.5 10.0 2.6 3.7 2.3 3.9 3.8 2.9 27 3.6 28.3 27.2 2.1 2.1 3.8 14.4 2.2 28 3.5 17.5 25.6 2.5 2.4 3.9 22.3 2.4 29 3.1 22.1 25.1 17.0 8.7 10.8 8.0 2.1 3.7 3.5 12.9 3.5 3.2 3.8 3.7 2.3 5FU 15.5 16.3 17.1 14.9 16.7 15.0 17.2 16.4 Table 20. Percentage of pyrimidine compounds against cells at TGI concentrations. Cytotoxicity values 29 % Cytotoxicity A549 Calu1 H1650 A172 C6 B35 Beas2B HC 31 2.4 11.8 6.9 3.1 3.4 20.0 29.5 2.5 32 3.4 2.9 3.1 3.5 3.9 3.4 20.8 2.5 33 3.8 29.2 3.9 3.2 10.8 12.9 2.6 5.4 34 2.8 28.1 24.1 3.3 3.7 3.0 24.7 3.5 35 3.0 29.0 27.2 18.3 2.6 11.5 3.1 3.8 36 3.0 5.5 2.7 3.5 10.9 67.1 2.7 3.0 37 2.9 3.8 2.7 3.6 2.5 3.1 3.7 3.5 38 3.6 3.1 3.6 3.7 2.8 3.9 3.8 3.5 39 3.9 2.9 3.7 3.9 2.4 3.8 3.2 3.4 40 3.7 27.8 3.3 21.4 3.7 25.3 2.6 2.7 41 3.6 28.0 3.0 3.3 3.8 3.8 24.2 2.5 5FU 15.5 16.3 17.1 14.9 16.7 15.0 17.2 16.4 a. Effect of Test Compounds on Cell Migration and ImageR Analysis The migration capacity of cells is a key feature of cancer development and new Pharmacological agents target cancer cells that have the capacity to migrate and undergo apoptosis. It can evade the mechanism. Therefore, newly developed pharmacological agents 5 One of the aims is to significantly reduce the migration capacity of cancer cells. According to migration tests conducted over time, the tested compounds are cancer-causing. reduced the migration capacity of the cells to a certain extent compared to the control cells It is understood. Table 21. Migration analysis results of compounds performed with ImageR. % Area A172 cell line (% Area) B35 cell line (% Area) Space Day 0 Space Day 1 Space Day 2 Space closing rate % Area Space Day 0 Space Day 1 Space Day 2 Space closing rate 16 52.15 71.05 80.73 -28.58 1 62.24 50.18 47.51 14.73 17 51.45 45.47 20.84 30.60 2 69.98 65.36 74.51 -4.53 18 51.34 78.04 84.18 -32.83 6 60.71 47.47 36.95 23.75 31 52.48 74.27 73.58 -21.10 7 74.83 55.27 39.11 35.72 32 41.98 74.85 70.09 -28.10 8 61.96 64.97 10.63 51.36 33 55.06 50.98 64.67 -9.61 10 72.47 39.82 26.92 45.55 34 53.80 53.51 56.83 -3.02 11 70.94 41.45 32.43 38.51 35 61.70 76.2 5 82.60 -20.90 12 68.42 42.95 36.73 31.68 37 49.56 72.81 59.54 -9.98 13 58.62 29.61 39.60 19.01 38 45.35 81.04 74.26 -28.90 19 70.66 65.26 62.58 8.08 39 52.27 83.98 60.22 -7.94 20 69.37 48.12 34.81 34.55 40 50.36 85.48 70.95 -20.59 21 80.02 46.82 19.11 60.91 41 52.18 83.69 72.40 -20.22 22 62.90 30.93 41.77 21.13 24 76.03 56.04 37.19 38.83 69.52 39.96 6.64 60.87 Control 75.58 16.52 0 75.58 Control 63.94 43.13 0 63.94 Table 22. Migration analysis results of compounds performed with ImageR. Compound HC cell line (% Area) Gap Day 0 Gap Day 1 Gap Day 2 Gap closing rate 3 81.25 36.16 34.43 46.81 9 85.93 75.53 68.46 17.46 4 57.13 35.85 21.54 35.58 51.24 45.65 44.16 7.07 14 77.76 68.28 44.27 33.49 59.77 67.10 43.68 16.08 23 88.20 47.45 33.43 54.76 26 65.21 16.32 0 65.21 27 67.13 40.65 32.99 34.13 28 65.45 26.48 11.01 54.43 29 55.18 58.08 61.45 -6.26 70.67 55.32 46.40 24.26 Control 90.77 11.22 0 90.77 Determination of DNA Banding Potentials of Compounds 1, 9, 16, and 19 5 The invention is based on the antiproliferative activity of compounds numbered 1, 9, 16 and 19. The existence of the apoptosis mechanism was investigated using the DNA laddering assay. For this purpose, Beas2B, C6, A172 and were treated with IC50 concentrations of the relevant compound. DNA was isolated from B35 cells, and the apoptotic effects of substances 1, 9, 16, and 19 on the cells were investigated. The effects were determined by observing the banding in the cell DNA. Figure 8 also shows 10. as seen, DNA isolated from control cells untreated with 1, 9, 16 and 19 In comparison, there is no obvious difference in DNA samples isolated from cells treated with the substance. Banding was observed. DNA banding test results for items 1, 9, 16 and 19. Its antiproliferative effect is most likely due to stimulating the apoptosis mechanism. It was interpreted as having accomplished something. 15 The mechanism of apoptosis based on the antiproliferative activity of compounds 12 and 26. The presence of the compound was investigated using the DNA laddering assay. For this purpose, the compound in question was... DNA from Beas2B, C6, A172, and B35 cells treated with IC50 concentrations. By isolating them, the apoptotic effects of substances 12 and 26 in the cell were investigated in 20 cells of the cell's DNA. This was determined by observing the banding. As seen in Figure 9, with 12 and 26 31 Compared to DNA isolated from untreated control cells, the substance A clear banding pattern was observed in DNA samples isolated from the treated cells. DNA The banding test results show that substances 12 and 26 have a very large antiproliferative effect. It was interpreted that this was likely achieved by stimulating the apoptosis mechanism. Topoisomerase I of Substances 33, 34, 35, 9, 10, 11, 22, 23, 24, 1, 2, 3, 16, 17 and 18 Determination of Inhibitory Activities Within the scope of the invention, articles 33, 34, 35, 9, 10, 11, 22, 23, 24, 1, 2, 3, 16, 17 and 18 are included. The effects on topoisomerase I activity were determined using the topoisomerase I inhibition test. According to the test results, items 33, 34, 35, 9, 10, 11, 22, 23, 24, 1, 2, 3, 16, 17 and 18 It halted the DNA relaxin activity of topoisomerase I (Figure 10). Based on these results, Substances 33, 34, 35, 9, 10, 11, 22, 23, 24, 1, 2, 3, 16, 17 and 18 have an anticancer effect. It can be said that topoisomerase I inhibition is responsible for this part. 15 The effects of compounds 12, 26, 27, 28, 19, 20, 21, 37 and 38 on topoisomerase I activity. The effects were determined by the topoisomerase I inhibition test. According to the test results, 12, 26, 27, 28, Substances 19, 20, 21, 37, and 38 inhibited the DNA-relaxing activity of topoisomerase I. (Figure 11). According to these results, substances 12, 26, 27, 28, 19, 20, 21, 37 and 38 have anticancer properties. It can be said that topoisomerase I inhibition is responsible for part of its effect. DNA / BSA Binding Results of Compounds 34, 35, 36, 9, 10 and 11 Evaluation Many of the anticancer or other effective drug molecules used today are pharmacological. 25 It sometimes exerts its effects by binding to DNA biomacromolecules. These molecules Its transport in the blood is also mostly via albumin protein (Human Serum Albumin, HSA). This Therefore, substances 34, 35, 36, 9, 10 and 11 bind to DNA / BSA (Bovine Serum Albumin). Their characteristics have been determined and are presented here. Figure 12 shows items 34, 35, 36, 9, 10, and 11. The maximum peaks of change observed with CT-DNA and BSA were 332, 342, 342, 256, 276, and 30, respectively. and were measured as 310 (for DNA), 330, 342, 346, 292, 294 and 292 (for BSA) nm. Figure As CT-DNA concentration increased in 12 (A), absorbance decreased (34, 35 and 36) or increased. (9, 10 and 11) are observed. This is called the hypochromic and hyperchromic effect, and the test substances It shows that it interacts with CT-DNA. As the BSA concentration increases, the absorbance increases as shown in Figure 12 (C). 32 It is observed that it either decreases (34, 35 and 36) or increases (9, 10 and 11). This is called hypochromic and This is called the hyperchromic effect and indicates that the test substances interact with BSA. This interaction is CT- It appears in DNA in the intercalation mode. Substances 34, 35, 36, 9, 10 and 11 are CT-DNA. The binding constants (Kb) with (Figure 12B) and BSA (Figure 12D) are 4.4x10³M¹־, respectively. 2.7x10⁴M, 1.7x10⁴M, 9.9x10³M, 1.7x10⁴M, and 4.8x10⁴M (for DNA), and 5 1.3x10⁴M,¹־ 3.5x10³M1.4 ,¹־x10⁴M,¹־ 1.6x10⁴M,¹־ 1.2x10⁴M¹־, and 1.2x10⁴M¹־ (for BSA) It has been measured as follows. Evaluation of DNA / BSA Binding Results of Compounds 27 and 28 The DNA / BSA (Bovine Serum Albumin) binding properties of substances 27 and 28 were determined. presented here. In Figure 13, molecules 27 and 28 are CT-DNA (Calf thymus DNA) (Figure 10 Interaction graphs with (Figure 13A) and BSA (Figure 13C) are shown. Figures 27 and 28 are shown in Figure 13. The maximum peaks where the changes in these substances occur with CT-DNA and BSA are 314 and 334, respectively. The measurements were 308 nm (for DNA) and 334 nm (for BSA). Figure 13 shows CT-DNA and BSA. It is observed that as the concentration increases, the absorbance decreases (27) or increases (28). This is called hypochromic and hyperchromic effect and refers to the interaction of test substances with CT-DNA and BSA. 15 This interaction appears in CT-DNA in the form of intercalation. Articles 27 and 28... The binding constants (Kb) with CT-DNA (Figure 13B) and BSA (Figure 13D) are as follows: The measurements were 1.0x10⁴M².9,¹־x10³M¹.2,¹־x10⁴M¹־ and 4.0x10³M¹־. Molecular Docking Studies of Invention Compounds 20 To determine ligand-target interactions in the investigation of compounds that may be potential drug candidates, and Molecular docking, an in silico method for interpretation, is frequently used. Protein data of synthesized compounds that experimentally showed effective anti-cancer activities The crystal structure obtained from its base was analyzed using the Schrödinger Maestro 2021-2 program. The results of the molecular docking study conducted in silico environment are presented in Table 23. 25 Molecular docking is used to examine the best results for outcomes studied as anticancer agents. An analysis has been conducted. Table 23. Results of binding parameter values ​​for the determined compounds. Compounds Docking score value XP gscore value ΔGBintegration value Glide energy value 12 -7.519 -7.519 -45.36 -38.887 19 -6.818 -6.818 -40.73 -35.955 -7.034 -7.034 -47.32 -38.231 21 -6.515 -6.515 -38.55 -37.481 26 -7.117 -7.118 -50.12 -38.860 27 -7.099 -7.099 -46.96 -36.700 33 28 -6.956 -6.958 -46.83 -37.700 34 -8.291 -8.291 -63.09 -52.592 35 -8.271 -8.271 -66.62 -49.183 36 -8.375 -8.375 -62.30 -48.380 37 -7.838 -7.839 -55.37 -44.852 38 -7.865 -7.865 -55.82 -45.576 10 20 30

Claims

34 REQUESTS 1. Pyralosin derivative is an anticancer compound with the following properties: {X-[5-(Y)-Z-1H-pyrazole-3- having the compound structure of [ilphenyl]amine; X is one of the values ​​2, 3, and 4, Y is “2,4-Difluorophenyl”, “3,5-Difluorophenyl”, “2-Chloro-6-5 One of the values ​​"fluorophenyl", "2-Chloro-5-fluorophenyl" and "2-Chloro-4-fluorophenyl" is Z; Having one of the values ​​"4,5-dihydro" and "empty" It is characteristic.

2. It is a compound according to claim 1, and its property is; {2-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazole- It is a compound of 3-yl]phenyl}amine, 10 Formula (22) It is expressed by formula (22).

3. It is a compound according to claim 1, and its property is; {3-[5-(2,4-Difluorophenyl)-4,5-dihydro-1H-pyrazole- It is a compound of 3-yl]phenyl}amine, 15 Formula (23) It is expressed by formula (23).

4. It is a compound according to claim 1, and its property is; {4-[5-(2,4-Difluorophenyl)-1H-pyrazole-3- [ilphenyl]amine compound, 20 Formula (24) It is expressed by formula (24). 35 5. It is a compound according to claim 1, and its property is; {2-[5-(3,5-Difluorophenyl)-4,5-dihydro-1H-pyrazole- It is a compound of 3-yl]phenyl}amine, Formula (25) 5 It is expressed by formula (25).

6. It is a compound according to claim 1, and its property is; {2-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H- It is a pyrazole-3-yl]phenyl}amine compound, Formula (26) 10 It is expressed by the formula (26).

7. It is a compound according to claim 1, and its property is; {3-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H- It is a pyrazole-3-yl]phenyl}amine compound, Formula (27) 15 It is expressed by formula (27).

8. It is a compound according to claim 1, and its property is; {4-[5-(2-Chloro-6-fluorophenyl)-4,5-dihydro-1H- It is a pyrazole-3-yl]phenyl}amine compound, Formula (28) 20 36 It is expressed by formula (28).

9. It is a compound according to claim 1, and its property is; {2-[5-(2-Chloro-5-fluorophenyl)-4,5-dihydro-1H- It is a pyrazole-3-yl]phenyl}amine compound, Formula (29) 5 It is expressed by the formula (29).

10. It is a compound according to claim 1, and its property is; {2-[5-(2-Chloro-4-fluorophenyl)-4,5-dihydro-1H- It is a pyrazole-3-yl]phenyl}amine amine compound, Formula (30) 10 It is expressed by formula (30). 20

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