Soy lecithin-based encapsulated curcumin and its production method for the treatment of prostate cancer.

Abstract

The invention relates to soy lecithin-based encapsulated curcumin, the method of producing this soy lecithin-based encapsulated curcumin, and its use in the treatment of prostate cancer. The invention demonstrates that soy lecithin-based encapsulated curcumin exerts a stronger toxic effect on cancer cells compared to free curcumin. Intracellular uptake studies also show that soy lecithin-based encapsulated curcumin accumulates five times more in cancer cells than the free form. Furthermore, according to animal experiments conducted on the 22Rv1 xenograft prostate model, the average tumor volume in the group treated with SLKUR was approximately 1 / 10 lower compared to the control group. These results demonstrate that SLKUR has a higher anti-tumor effect than KUR.The invention provides a new formulation of curcumin, containing 100% natural SLKUR and a highly water-soluble carrier that enhances its absorption, enabling its use as an anti-cancer agent.

Description

1 TARIFF SOY LECITHIN-BASED ENCAPSULATE FOR THE TREATMENT OF PROSTATE CANCER. CURCUMIN AND THE PRODUCTION METHOD OF THIS ENCAPSULATED CURCUMIN Technical Area 5 The invention is based on soy lecithin and is intended for use in the treatment of prostate cancer. This invention relates to curcumin (SLKUR) and the method of producing this formulation. The subject of the invention is SLKUR. an adult male with adenocarcinoma compared to the free curcumin form (CUR). on an epithelial cell line (TRAMP-C2) isolated from the prostate of a transgenic mouse It shows a higher toxic effect. In addition, the 10 mentioned in the cell uptake experiments SLKUR is taken up by TRAMP-C2 cells five times more than KUR. State of the Art Cancer occurs when body cells grow and divide uncontrollably, leading to abnormal changes. It forms cell clusters, sometimes spreading to other tissues and leading to metastasis. 15 It is a group of diseases caused by cancer. Cancer usually develops intracellularly. genetic mutations and epigenetic changes occurring in mechanisms This occurs as a result of mutations in tumor suppressor genes. inactivation, activation of proto-oncogenes, or DNA repair It may be related to disruptions in their mechanisms. Cancer triggers 20 among them are chemical substances (carcinogens), radiation, viral infections, and genetics. There is a predisposition. Active ingredients used in cancer treatment, cancer It varies depending on the type and stage. The most commonly used treatment methods are: among them chemotherapy (cytotoxic drugs), immunotherapy, targeted therapies and Radiotherapy is included. Chemotherapy includes 25 drugs such as doxorubicin, cisplatin, and paclitaxel. Cytotoxic agents are used. Trastuzumab is used in targeted therapies. Monoclonal antibodies or kinase inhibitors are preferred in immunotherapies. Immune checkpoint inhibitors (such as PD-1 and CTLA-4) are widely used. Drug delivery systems aim to increase the therapeutic efficacy of anti-cancer agents and minimize side effects. It is of great importance to reduce their effects. These systems target drugs to the target. to deliver, improve bioavailability and minimize systemic toxicity It has been designed. In this context, phytochemicals with strong anti-tumor properties, 2 They have emerged as promising candidates for cancer treatment. Multiple natural products Their broad-spectrum activity on signaling pathways means these compounds are effective in abnormal cells. proliferation, apoptosis regulation disorder, angiogenesis, invasion, and metastasis, etc. It has a unique advantage because it disrupts key processes in cancer development. It offers. In addition, the diversity of phytochemicals, their structural complexity, and their affordability, 5 Their low toxicity and intrinsic biological activity make them ideal for therapeutic use. In addition, natural products create synergy with traditional chemotherapy and radiotherapy. It has been shown that they have the ability to create and potentially increase their effectiveness, while also having side effects. It reduces the effects. Integration of phytochemicals into advanced drug delivery systems. This opens up new opportunities to develop effective and multi-targeted cancer therapies. 10 Curcumin is a naturally occurring polyphenol found in turmeric, with anticancer and anti-inflammatory properties. Its antioxidant properties have been widely studied scientifically. However, Curcumin has low bioavailability and limited stability in circulation, This limits the therapeutic efficacy of this bioactive molecule. Curcumin has a carrier 15 Encapsulating it provides a solution to these problems, offering an effective treatment option. This method aims to increase the bioavailability of curcumin and its circulation. to ensure longer stability in the system and targeted treatment effectiveness It offers versatile advantages for highly optimizing the system. Inside the carrier Encapsulated curcumin has higher stability in the gastrointestinal system. and bioavailability is significantly increased. In addition, carrier systems By enabling the specific transport of curcumin to the tumor microenvironment, it promotes healthy growth. It minimizes toxic effects on tissues. Controlled release mechanism. Thanks to this, the therapeutic effect of encapsulated curcumin can be prolonged. and can optimize the efficacy profile of the drug. However, encapsulated curcumin 25 Their formulations also have some disadvantages. Production processes Complexity and high costs are major limitations on the widespread use of these systems. This is one of the factors. Additionally, the targeting capabilities of carrier systems are insufficient. In this case, treatment effectiveness may be limited. Metabolism and elimination. During these processes, there is a risk of carriers accumulating in the liver and spleen, which is 30 It can reduce the systemic effect of curcumin. Lack of clinical data also makes encapsulation problematic. to fully evaluate the long-term safety and efficacy of curcumin This makes things more difficult. As a solution to these problems, targeted intelligent carrier systems are being developed. development, use of combination therapies with other chemotherapeutic agents, and 3 Pharmacokinetic optimizations are recommended. New PEG-enhanced systems and... Formulations developed with biodegradable materials improve the distribution of curcumin in tissues. and can improve its bioavailability. Furthermore, it enhances the safety of encapsulated curcumin. And extensive clinical studies supporting its effectiveness are needed. In conclusion, encapsulated curcumin is a promising approach in cancer treatment. 5 While it offers these benefits, its therapeutic effectiveness can be further improved by addressing technical and clinical deficiencies. It can be increased. This was done by Diomede and his colleagues, at the known state of the art. In the study, curcumin (CurLIP) encapsulated using a carrier system was 10 It discusses a study that explores the potential of suppressing inflammation and Effects on human stem cells in the context of periodontal diseases is being investigated [1]. Due to the anti-inflammatory properties of the carrier system in question a potential treatment method for inflammation-related diseases, such as cancer. It is stated that this is possible, but no evidence is presented to support this. 15 Disadvantages of carriers include low stability, limited drug loading capacity, and high production costs, challenges encountered in increasing targeting effectiveness, and The need to optimize bioavailability is included. The study focuses on encapsulated... Although it emphasizes that processed curcumin is effective in reducing inflammation, it also highlights cancer. It has shortcomings in terms of its applicability to treatment. 20 Another study by Bala and colleagues, which is part of the known state of the art... In this study, polymer-based carrier systems were used to achieve controlled and targeted release of curcumin. It has been shown that it increases its effectiveness by providing [2]. In particular, PLGA (polylactic-co- The cellular uptake of curcumin encapsulated in (glycolic acid) nanoparticles was 25%. It has been determined that the effect has increased and the anti-cancer effects have been strengthened. These formulations, cancer It has been found to be effective in reducing proliferation in cells and inducing apoptosis. However, these systems present high costs and scaling challenges in their production processes. These are among the important limitations. Another study by Sharma and colleagues, which is part of the known state of the technique... According to one study, chitosan nanoparticles have biocompatible and non-toxic structures. Chitosan significantly increases the oral bioavailability of curcumin [3]. systems based on the stability and absorption of curcumin, especially in the gastrointestinal system 4 It has shown effective therapeutic potential by improving [condition]. However, chitosan Long-term stability of systems and comprehensive toxicological studies for clinical applications. Their assessments are not yet sufficient. Another study within the known state of the art, Mukherjee et al. 5 According to the study conducted, solid lipid nanoparticles (SLNs) have high stability. And thanks to its biocompatible structures, curcumin's circulating time is extended and it is effective against cancer. It has been reported that it is delivered to targeted cells [4]. In both in vitro and in vivo models. Curcumin, when applied to SLNs, enhanced their anti-cancer and anti-inflammatory effects. However, SLN systems have limited drug delivery capacity and formulation 10 Its stability decreasing under certain conditions is a significant disadvantage. Another study by Wang and colleagues, which is part of the known state of the technique... According to the study, polymeric microspheres have the capacity to provide controlled release. It increases the systemic bioavailability of curcumin [5]. Especially alginate and PLGA 15 Microspheres based on a microsphere base increase solubility in the gastrointestinal system, thereby enhancing curcumin's anti- It enhanced its proliferative and pro-apoptotic effects in cancer cells. However, repeatability and large scale of methods used in the production of microspheres The challenges encountered in production are noteworthy. In another study within the known state of the art, Chen et al. According to the research, hydrogel-based systems are effective in transporting curcumin in three ways. It offers three-dimensional water-retaining structures [6]. Curcumin-loaded hydrogel systems, in particular By providing high stability and long-lasting release in topical and local applications, it offers anti- These systems have enhanced inflammatory effects. 25 on skin and mucosal surfaces It enhances the effectiveness of curcumin. However, the mechanical strength of hydrogels and Biocompatibility needs to be optimized for different applications. According to another study within the known state of the art, Ranjan and colleagues Curcumin-loaded nanofibers prepared by electrospinning method have a large surface area of ​​30 and, thanks to its targeted release properties, it is specifically delivered to cancer cells. It has been determined that [7]. These transporter systems are an important factor in the anti-cancer effects of curcumin. This has led to improvements in the production processes of nanofiber systems. The technical complexity and high costs prevent the widespread use of these systems. These are limiting factors. Studies involving current techniques encapsulate via carrier systems. Curcumin has 5% greater solubility, stability, and bioavailability compared to free-form curcumin. This shows that it offers significant advantages in terms of polymeric nanoparticles, solids. Lipid nanoparticles, microspheres, hydrogel-based systems, and nanofiber formulations, By enhancing the anti-cancer, anti-inflammatory, and pro-apoptotic effects of curcumin, clinically It shows promising results for its use. However, in most of these systems... 10 issues such as scaling, production costs, long-term stability and biocompatibility There are shortcomings. Addressing these shortcomings will enable the use of carriers in clinical applications. This will further enhance the effectiveness of curcumin systems. The limitations and inadequacies of current technological solutions, encapsulated curcumin The lack of sufficient studies on the use of these structures in cancer treatment, 15 Low stability, limited drug loading capacity, production difficulties, high production cost. and due to reasons such as low targeting effectiveness, there is a need for a solution in this area. Improvements are needed. Brief Description and Objectives of the Invention 20 The invention describes soy lecithin-based encapsulated curcumin (SLKUR) and this formulation. The production method of encapsulated curcumin is described. This SLKUR is used for prostate problems. Anti-tumor effects on cancer in an in vivo model of an adult with adenocarcinoma An epithelial cell line (TRAMP-C2) isolated from the prostate of a male transgenic mouse. It was tested on mice using cytotoxicity experiments. Additionally, during these studies, 25 mice were examined. Fibroblast cell line (Swiss 3T3) was used as a control group. The obtained According to the data, SLKUR is more effective in TRAMP-C2 cells compared to free curcumin (KUR). It exhibits excessive toxic effects. Intracellular uptake studies, however, reveal that the invention concerns SLKUR. This proves that it is taken up into the cell 5 times more than KUR. The main purpose of the invention is to develop SLKUR for use in the treatment of prostate cancer. This is achieved thanks to its ability to increase the low bioavailability of curcumin. The resulting SLKUR formulation has an effect on the bioavailability and cellular activity of curcumin. It shows positive effects. This formulation effectively targets the target cells. 6 by both reaching and reducing the side effects of traditional treatment methods It offers a safe and effective treatment option. The effectiveness of the SLKUR formulation is demonstrated by both in vitro and in vitro using the 22Rv1 xenograft prostate cancer model It has been tested in in vivo experiments. This model, which mimics the biology of prostate cancer, Experiments have proven it to be an ideal choice, and SLKUR treatment in tumor volume is 5. A tenfold greater reduction in size was observed compared to the post-control. These experiments... As a result, SLKUR effectively inhibited cell proliferation and showed potent results. It has been shown to exhibit anti-tumor activity. SLKUR formulation with 100% natural ingredients, This was developed for the first time in the literature, and its anti-cancer effect was also demonstrated in the 22Rv1 in vivo model. This study has been examined for the first time in the literature. This innovative study investigated encapsulated curcumin in prostate 10 It brings a fresh perspective to the literature by making a significant contribution to its use against cancer. This invention, SLKUR, has resulted in an alternative to traditional prostate cancer treatment. reducing the side effects of the methods, and providing a more effective and safer treatment option. It is presented. Explanation of the Figures Figure 1: Particle size distribution of SLKUR. Figure 2: FT-IR results of KUR and SLKUR formulations. (a: curcumin, b: soy) lecithin and c: soy lecithin-based encapsulated curcumin) Figure 3: Cytotoxicity of SLKUR on human epithelial prostate cell PNT1A and human prostate 20 Cytotoxicity results of cancer cells against 22Rv1 and LNCaP cell lines (A: PNT1A and SLKUR, B: 22RV1 and SLKUR, C: LNCaP and SLKUR, D: PNT1A and KUR, E:22RV1 and KUR, F: LNCaP and KUR) Figure 4: PNT1A, 22Rv1 and LNCaP cell lines treated with SLKUR and KUR. intracellular fluorescence intensity change coefficients (A: SLKUR cell recruitment and B: KUR 25 (cell procurement) Figure 5: SLKUR-induced apoptotic cell death (A: 22Rv1 and B: LNCaP) Bright field and fluorescence images of cells and C: 22Rv1 and LNCaP in graph. (Analysis results of average fluorescence intensity for cells) Figure 6: Western blot 30 of SLKUR and KUR application at PNT1A, 22Rv1 and LNCaP. Results (A: Protein samples, truncated caspase-9 (c-Cas 9) and PARP apoptotic Blotting results for markers and B: western regions of c-PARP and c-Cas 9 expression. (blot analysis results) 7 Detailed Description of the Invention The invention relates to soy lecithin-based encapsulated curcumin (SLKUR) and its production method. This is related. The encapsulated curcumin in question has been shown to have therapeutic effects on prostate cancer. This has been proven in vivo. In cytotoxicity analyses, transgenic mouse prostate The adenocarcinoma cell line TRAMP-C2 was used, and Swiss 5 was used as a control cell line. 3T3 mouse fibroblast cells were selected, and the results showed that encapsulated curcumin was released. Significantly higher toxic effect on TRAMP-C2 cells compared to the other form. In addition, intracellular uptake experiments show that the SLKUR formulation... TRAMP-C2 cells show that it is taken up 5 times more than KUR. Soy lecithin-based encapsulated for use in prostate cancer treatment. curcumin. The FTIR spectrum of the encapsulated curcumin, which is the subject of the invention, shows the aromatic moiety C=C at 1627 cm-1. Tension, benzene ring tension vibration at 1589 cm−1 and C=O and C=C 15 at 1516 cm−1 It includes vibrational peaks. These peaks are characteristic peaks of curcumin and are characteristic of curcumin. This shows the presence of nanocarriers containing curcumin in nanocarriers. The peaks that appear when encapsulated indicate an interaction between curcumin and lecithin. This indicates its presence. In the spectrum of curcumin-containing nanocarriers, it is 3508 cm⁻¹. The disappearance of the peak is most likely due to the hydrogen bond 20 of the phenolic –OH group of curcumin. It shows that it interacts with lecithin via this pathway. Main absorption peaks of curcumin weakened and shifted peaks when encapsulated in nanocarriers; this is also true for curcumin. This suggests that their molecules are located within nanocarriers. 25 encapsulated curcumin for use in the treatment of prostate cancer. synthesis method; i. For the preparation of soy lecithin dispersion, sterile soy lecithin is used. Mixing with injection water in a homogenizer, ii. In order to reduce the particle size of the resulting dispersion, type Performing the procedure using a sonicator device, 30 iii. The dispersion is freeze-dried and powdered using a lyophilizer. bringing, 8 iv. the powder is left to stand in a vacuum oven and then tightly sealed. Soy lecithin should be stored in containers in the refrigerator. obtaining nanoparticles, v. Sterile preparation of soy lecithin nanoparticles and pure curcumin. Mixing with water for injection, 5 vi. The mixing process is carried out using a type sonicator. vii. the resulting soy lecithin-based encapsulated curcumin dispersion Drying after freezing using a lyophilizer, viii. then, once again mixed with a sonicator under the same processing conditions processing, 10 ix. Re-frozen soy lecithin-based encapsulated curcumin re-drying of the dispersion with a lyophilizer, x. The resulting soy lecithin-based encapsulated curcumin was placed in a vacuum oven. After being left to stand, they are stored in tightly sealed containers in the refrigerator. storage 15 It includes the steps involved in the process. An application of the synthesis method described in the invention; i. To prepare a 2% (weight / volume) soy lecithin dispersion, use soybean 20 Lecithin was injected with sterile water for injection for 15-30 minutes at 21,000 rpm. mixing in a homogenizer of a certain value, ii. In order to reduce the particle size of the resulting dispersion, type Using a sonicator device, 5 each at 70 kHz power and 70% vibration. The transaction lasted a total of 15 minutes, with three 25-minute intervals. to be done, iii. dispersion, using a lyophilizer, at -50 °C and 0.1 millibar pressure. freeze-dried and powdered, iv. The powder is left in a vacuum oven for 1 hour and then tightly sealed. Soy lecithin can be stored in sealed containers in the refrigerator for 30 days. obtaining nanoparticles, v. Prepared soy lecithin nanoparticles and pure curcumin, in a 3:1 ratio. weighed and mixed with sterile water for injection, 9 vi. Mixing process using a type sonicator at 70 kHz power and 70% vibration, to be performed at 1 / 2 minute intervals for a total of 45 minutes, vii. the resulting soy lecithin-based encapsulated curcumin dispersion After freezing, it is heated to -50 °C and 0.1 millibar using a lyophilizer. drying under pressure, 5 viii. then, once again for 45 minutes under the same operating conditions (5 (at minute intervals, with a sonicator at 70 kHz power and 70% vibration) processed by mixing, ix. re-frozen soy lecithin-based encapsulated curcumin dispersion, Re-drying with a lyophilizer at -50 °C and 0.1 millibar pressure, 10 x. The resulting soy lecithin-based encapsulated curcumin was placed in a vacuum oven for 1 hour. After being left to stand for a certain period, they are placed in tightly sealed containers in the refrigerator. hiding It includes the steps involved in the process. The invention describes the production of soy lecithin-based encapsulated curcumin by sonication and cryopreservation. Drying methods are applied. First, soybeans are dried at a rate of 2% (weight / volume). A lecithin dispersion is being prepared. During this process, soy lecithin is sterilized. Injected with water for 15-30 minutes in a high-speed homogenizer (21000 20 (rpm, Hidolf, Germany) are mixed. Then, the particles of the resulting dispersion are mixed. To reduce its size, a type sonicator device was used at 70 kHz power and 70% In vibration mode, the process lasts for a total of 15 minutes, with three 5-minute intervals. Particle size and Zeta Potential measurements are performed using Dynamic Light Scattering. After being performed with the (DLS) device, dispersion is carried out at -50 °C 25 with the help of a lyophilizer. It is freeze-dried under temperature and 0.1 millibar pressure. It is then reduced to powder. The product is placed in a vacuum oven for 1 hour and then tightly sealed. The prepared soy lecithin nanoparticles are stored in containers in the refrigerator. Pure curcumin is weighed and mixed with sterile water for injection in a 3:1 ratio. The mixing process was carried out using a type sonicator at 70 kHz power and 70% vibration for 5 minutes (30 seconds). This is carried out intermittently for a total of 45 minutes. The resulting soy lecithin Curcumin-based encapsulated dispersion, after freezing, processed using a lyophilizer - It is dried at a temperature of 50 °C and a pressure of 0.1 millibars. Then, the same process is repeated. under these conditions once again for 45 minutes (in 5-minute intervals, 70 kHz power and 70%). (in vibration) it is processed by mixing with a sonicator. Finally, again The frozen dispersion is lyophilized at -50 °C and 0.1 millibar pressure. It is dried again. The resulting soy lecithin-based encapsulated curcumin is vacuum-packed. After being left in the oven for 1 hour, they are placed in tightly sealed containers and refrigerated. It is hidden. 5 Dynamic Light Scattering in the Characterization of Soy Lecithin-Based Encapsulated Curcumin Average size, zeta potential, and polydispersity index were determined using a (DLS) device. (PDI) values ​​have been determined. The DLS method was used to analyze materials at nano and micron dimensions. light, which allows us to understand the stability and dynamic properties of its particles. It is a technique limited by wavelength and provides information about material properties "at long wavelengths". It provides information. Zeta potential measurements of a dispersion or emulsion This is an important indicator in evaluating the stability of colloidal drug carriers. to analyze the surface properties of structures such as systems and their stability in different environments It is performed to predict behavior. In general, zeta potential The higher the magnitude of the value, the greater the dispersion or emulsion's 15. This means that its stability is so good. In terms of stability, the load... Whether it is positive or negative makes no difference. The particle size of SLKUR is Dynamic. The measurements were evaluated using a Light Scattering (DLS) instrument. The measurements show that SLKUR's particles... Its size is approximately 120 nm and its Polydispersity Index (PDI) value is 0.3 This shows that (Figure 1). Each graph in Figure 1, for example, is 20 It represents three independent, different measurements. The proposed size for nanoparticle dispersion. The range is 10-200 nm. PDI values ​​can vary between 0.0 and 1.0. From 0.05 Small values ​​generally indicate uniformly distributed samples, while values ​​above 0.7 The values ​​indicate highly distributed systems. In pharmaceutical delivery applications, A PDI below 0.4 is considered an acceptable characteristic. Also, 25 PDI values ​​below 0.5 indicate a homogeneous and narrow size distribution. This The results demonstrate that the nanoparticles in question have a homogeneous structure. In addition, molecular bonds were characterized using FT-IR spectroscopy. Fourier transform infrared (FT-IR) spectroscopy reveals the bonds in molecular structures. For the purpose of definition, various 30 disciplines such as basic sciences, health sciences and engineering It is a method widely used in various fields. The basic principle of this technique is that molecules... infrared rays directed onto the bonds cause the bonds to vibrate and rotate. The result is based on absorption. FTIR results are given in Figure 2. Figure As can be seen from 2, at 3508 cm−1 (phenolic OH stretching vibration), at 1628 cm−1 11 (aromatic part C=C stretching), at 1597 cm−1 (benzene ring stretching vibrations), At 1509 cm−1 (C=O and C=C vibrations), at 1428 cm−1 (olefinic CH bending) vibrations), at 1278 cm−1 (aromatic C–O stretching vibrations), at 1024 cm−1 (C–O–C (stretch vibrations) signature peaks are shown. Typical peaks of lecithin are at 2926 and 2855. 1737 cm−1 (CH2 stretching vibration), 1648 cm−1 (symmetric C=O stretching vibration). at cm−1 (water shear band) and 1249 cm−1 (PO4 antisymmetric tension bands) This has been shown. When curcumin is incorporated into nanocarriers, the peak position and shape are similar to those of lecithin. At 2926 cm−1 and 2855 cm−1 (CH2 vibration absorption), at 1737 cm−1 (symmetric C=O stress vibration absorption) and similar at 1249 cm−1 (PO4 antisymmetric strain bands) It is observed here that curcumin-containing nanocarriers are present at 1627 cm−1 (aromatic part 10 C=C stress), at 1589 cm−1 (benzene ring stress vibrations) and at 1516 cm−1 It exhibited peaks (C=O and C=C vibrations), which are characteristic peaks of curcumin. and demonstrates the presence of nanocarriers containing curcumin. Curcumin The peaks that appear when encapsulated in nanocarriers are between those of curcumin and lecithin. This indicates the existence of an interaction. 15 in the spectrum of curcumin-containing nanocarriers. The disappearance of the 3508 cm⁻¹ peak most likely indicates the loss of the phenolic –OH group of curcumin. It has been shown to interact with lecithin via hydrogen bonding. The main component of curcumin... Absorption peaks were weakened and shifted when the peaks were encapsulated in nanocarriers; this indicates that This suggests that curcumin molecules are present within nanocarriers (Figure). 2). 20 The invention concerns a soy lecithin-based encapsulated curcumin formulation. For cytotoxicity studies, samples were obtained from the American Tissue Culture Collection (ATCC). The obtained cell lines were used. In the analysis of cell viability, the research The Cell Viability Test (WST-1) optimized by our group was applied. This 25 test, NAD(P)H-dependent dehydrogenase enzymes in metabolically active cells the principle of converting tetrazolium salt into a formazan group dye It is based on tetrazolium salt formazan as long as the cells remain alive. Since it is converted, the amount of formazan formed is directly related to metabolically active cells. It is related to the number. WST-1 analysis showed that SLKUR 30 was present at concentrations of 6.25, 12.5 and 25 μg / mL. and with 2.08, 4.14, 6.33 µg / mL DUR concentrations for 24, 48 and 72 hours. The treated prostate normal epithelial cells (PNT1A), 22Rv1, and prostate cancer cells This was done to determine the viability rate in the lines (22Rv1 and LNCaP). Whole cell The lines were standardized with 2.5x10³, 5x10³, and 7.5x10³ per groove, respectively. 12 They were seeded into 96-well plates at cell densities of 10x10³, 15x10³, and 20x10³. In the experiments, each cell line was placed in 96 wells, with 5000 cells per well. The plates were seeded and incubated in an incubator containing 5% (volume / volume) carbon dioxide at 37°C. They were incubated overnight. The next day, the cells were analyzed at a concentration of 6.25-25 μg / mL. Treatment with SLKUR at concentrations of 2.08-8.33 μg / mL and KUR at concentrations of 2.08-8.33 μg / mL for 24, 48 and 72 hours. was subjected to this. At the specified time points, cell culture medium with 10% The mixture, prepared to contain WST-1 reagent by volume / volume, is 100 for each well. Added in μL in an incubator containing 5% (volume / volume) carbon dioxide, It was incubated at 37°C in a dark environment for 1 hour. After incubation, Absorbance measurements were performed using a spectrophotometer at a wavelength of 450 nm. 10 At each time point, cell viability was higher than that of untreated control cells. The calculation was made assuming the obtained absorbance value is 100%. The aforementioned... The cytotoxic effects of different doses of SLKUR were investigated using PNT1A, 22Rv1, and LNCaP. PNT1A, 22Rv1 and LNCaP treated with SLKUR and KUR were analyzed (Figure 4). Based on the WST-1 cell viability test results of the cell lines, the cell numbers are 15. It was determined that in PNT1A cells, SLKUR and KUR were used for 24, 48 and 72 hours. the applications did not show a cytotoxic effect at any dose or time point This was observed. In contrast, in the 22Rv1 and LNCaP cell lines, 25 for 72 hours A significant cytotoxic effect was observed after treatment with μg / mL SLKUR. As a result of this treatment, the cell counts were measured as 950 and 2100, respectively. 20 However, treatments with free curcumin did not show a significant cytotoxic effect. No observations were made. After 72 hours of treatment with 8.33 μg / mL free curcumin, 22Rv1 The cell numbers in the LNCaP cell lines were determined to be 6600 and 6840, respectively. It has been done. Intracellular uptake of the subject of the invention, SLKUR, in PNT1A, 22Rv1 and LNCaP cell lines, by measuring the fluorescence intensity of curcumin accumulated within the cell was evaluated. For the experiment, SLKUR and SLKUR were used in PNT1A, 22Rv1 and LNCaP cells. By evaluating the cellular uptake of KUR, cells were injected with 25 µg / mL SLKUR for 48 hours. The cells were treated with 8.33 µg / mL KUR. After treatment, the cells were fixed and left for 30 minutes. It was stained for fluorescent imaging. The fixation process took 15 minutes at room temperature. throughout using 4% paraformaldehyde (PFA) in phosphate-buffered serum (PBS) This was performed. Following fixation, the cells try to get rid of the remaining fixative. 13 The images were washed three times with PBS. Nuclei were labeled with DAPI (blue). The images are... This was obtained using a fluorescence microscope with appropriate filters (Figure 3A). To further determine SLKUR within the cells, the cells were analyzed using a different method, 6 The cells were seeded onto well plates with 300,000 cells in each well, and a 5 overnight in an incubator environment containing 5% (volume / volume) carbon dioxide at 37°C. They were incubated. The next day, the cells were treated with 25 μg / mL SLKUR and 8.33 μg / mL for 4 hours. KUR was applied. After the application, the cells were collected and the resulting pellets were analyzed. a hypotonic solution (10 mM 4-(2-Hydroxyethyl)-1-piperazinetan sulfonic acid) (HEPES), 0.1 mM Ethylenediaminetetraacetic acid (EDTA), 0.1 mM Ethyleneglycol-bis(β-10 aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 0.1 mM Dithiothreitol (DTT), 50 mM Sodium fluoride (NaF), 1 mM Sodium orthovanadate (Na₃VO₄), 0.5 mM Phenylmethylsulfonyl Fluoride (PMSF), 30 mM β-glycerophosphate, 10 μg / mL Aprotin, 10 μg / mL Leupeptin, 10 μg / mL Pepstatin and deionized water (dH₂O) were added. After this process, the samples were vortexed. and left on ice for 10 minutes. After incubation, 1% of the cells were reduced to 15%. Nonidet NP-40 was added to each sample to reach the final concentration (volume / volume). and incubated again on ice for 10 minutes. Fluorescence of curcumin in the cells. Intensity measured at 420 nm excitation and 550 nm emission in the Varioskan Lux spectrophotometer. The invention was evaluated by measurement at the wavelength. The invention is treated with SLKUR and KUR. Changes in intracellular fluorescence intensity of PNT1A, 22Rv1 and LNCaP cell lines obtained 20 The coefficients were analyzed (Figure 3B). Fluorescence measured for the cellular uptake experiment. density values ​​compared to SLKUR-untreated cells representing the control group. Normalized. Fluorescence intensity measurements were taken at 420 nm excitation and 550 nm. The analysis was carried out at different emission wavelengths. According to the analysis results, SLKUR's KUR Compared to its previous form, it increased the uptake efficiency of cancer cells by 15 times. It has been determined that this formulation also has high targeting in cancer cell lines. It demonstrated its effectiveness and entered the cell more effectively compared to normal cells. It has been determined that it was taken. To determine apoptotic cell death in 22Rv1 and LNCaP cell lines, 30 TUNEL (Terminal Deoxynucleotidyl) in cells treated with SLKUR, which is the subject of the invention Transferaz dUTP Nick End Tagging) analysis was performed. TUNEL method, 3′-hydroxyl ends of double-stranded DNA breaks formed during apoptosis It is a technique that allows the detection of DNA fragmentation through labeling. 14 In the experiment, cells were seeded onto 8-well slides at a density of 50,000 cells / well, and 25 μg / mL SLKUR for 48 hours or 5 μg / mL for 90 minutes as a positive control The cell media was treated with μg / mL staurosporin (apoptosis inducer). After collection, adherent cells on the slide were 4% (volume / volume). It was fixed with paraformaldehyde solution and incubated at 37 °C for 1 hour. 5 Following incubation, the slides were washed with 1X Phosphate Buffered Saline Solution (PBS). and left on ice for 2 minutes for permeabilization. Then again 1X PBS The samples were washed, air-dried, and mixed with 50 μL of TUNEL reaction solution. It was applied and incubated at 37°C for 75 minutes. After incubation, TUNEL The reaction solution was removed and the samples were examined under a fluorescence microscope (515-565 nm / green filter) 10 22Rv1 and LNCaP cell lines were analyzed with staurosporin, SLKUR and KUR. images under light microscope and fluorescence microscope after treatment recorded (Figure 5). For these recordings, 25 µg / mL SLKUR was applied for 48 hours and 8.33 µg / mL DUR was applied. These images were taken at 20X under bright field and fluorescence. It was obtained with an objective lens and a scale value of 200 µm. In the graph shown in Figure 5C, 22Rv1 15 The results of the average fluorescence intensity analysis for LNCaP cells are shown. PK is a positive result in which apoptosis is induced with an apoptosis-inducing agent such as Staurosporin. This means control; this was determined by the TUNNEL test. Positive control. for 90 minutes with 5 μg / mL staurosporin and for 48 hours with 25 μg / mL SLKUR. Treated cells were examined using both light microscopy and fluorescence microscopy at a 20X objective. It was viewed with a microscope (Figure 5A). Obtained in fluorescence microscopy. In the analysis of the images, 22Rv1 and LNCaP prostate cells treated with SLKUR were found to be affected. apoptotic cell populations in cancer cell lines, normal prostate cell line It was found to be significantly increased compared to PNT1A (Figure 5B). In the images The observed radiation indicates that SLKUR induces apoptotic death in prostate cancer cells. 25 It clearly demonstrates this. The invention concerns apoptotic proteins in cells treated with SLKUR and KUR. Western blot analysis was performed to determine the levels. This analysis showed a level of 25 µg / mL. PNT1A, 22Rv1 and 30 treated with SLKUR and 8.33 µg / mL KUR for 48 hours This was performed on proteins isolated from LNCaP cells. The cells, They were seeded into 6-well plates with 300,000 cells per well, and It was treated with the specified curcumin concentrations. After the treatment... 30 µL RIPA buffer (1 mM PMSF, 0.5% non-ionic detergent) was added to the cells. (idet), 0.1% SDS, 1 mM NaF, 1 mM Na₃VO₄ and a protease inhibitor cocktail were added, The cell surface was scraped, the membrane was disrupted, and cell lysates were obtained. Protein The amount was measured using the Lowry method. The obtained protein lysates were incubated at 95°C for 5 minutes. It was denatured in Laemmli buffer throughout. Proteins in the cell homogenate (50 µg / well), sodium dodecyl sulfate 5 with concentrations ranging from 8% to 12%. Polyacrylamide was separated on gel electrophoresis (SDS-PAGE) gels. The separated gels... Proteins were mixed using the wet-transfer method at a current of 175 mA for 1.5 hours. Nitrocellulose was transferred to a membrane. The membranes were made of tris-polyoxyethylene containing 5% milk powder. sorbitan monolaurate (Tris-Tween) (100 mM Tris(hydroxymethyl)aminomethane hydrochloride buffer (Tris-HCl), 0.9% sodium chloride (NaCl), 0.05% polyoxyethylene sorbitan 10 The specimens were blocked in monolaurate-20 (Tween-20), pH 7.4 solution for 1 hour. The blocked membranes were incubated with appropriate primary antibodies at 4°C for 16 hours. The antibodies used were at the 5% concentration recommended by the purchasing company. The membranes were prepared in a Tris-Tween solution containing milk powder. The membranes were washed three times for 5 minutes. Tris-Tween 15 containing 5% milk powder, after washing with Tris-Tween solution. HRP (Horse Radish) diluted in solution at a ratio of 1:5000 or 1:10000 peroxidase) labeled anti-mouse antibody for 2 hours at room temperature They were incubated. Following incubation, the membranes were subjected to Tris-Tween heat treatment three times for 5 minutes. washed with solution - Horseradish Peroxidase (ECL-HRP) substrate solution They were kept in a waiting period. Finally, the membranes were processed in the ChemiDoc XRS+ (BioRad) device for 20 minutes. The invention, SLKUR and KUR, was examined and analyzed. PNT1A, 22Rv1 and The effect of LNCaP on apoptotic mechanisms of cell lines, western blot analysis. It was evaluated with SLKUR and KUR implementations PNT1A, 22Rv1 and LNCaP. Western impregnation results showing the effect of western infiltration on the apoptotic mechanism of cells. This is shown in Figure 6. In the analysis, 25 µg / mL SLKUR and 8.33 µg / mL 25 were administered for 48 hours. Evaluation of apoptosis in PNT1A, 22Rv1 and LNCaP cells using KUR, western ultrasound This was performed using the blot method. Protein samples were analyzed using truncated caspase-9 (c-Cas 9) and Apoptotic markers such as PARP were examined by blotting (Figure 6A). Presented Blots were obtained from three independent experiments, and β-actin was used for analysis. c- Representative densitometry of western blot analysis of PARP and c-Cas 9 expression 30 The graph is shown (Figure 6B). B-Actin expression was used as a control. c- Densitometric analysis of PARP and c-Cas 9 expressions was performed using Image J software. This analysis was performed and normalized according to the control level. Within the scope of this analysis, Cells were treated with 25 µg / mL SLKUR and 8.33 µg / mL free curcumin for 48 hours. 16 It was treated. PARP (116 and 89 kDa), which are involved in the apoptotic mechanism, and Expression levels of caspase 9 (48 and 35 kDa) proteins were determined by western blot method. Protein lysates were determined. They were incubated in Laemmli buffer at 95°C for 5 minutes. After denatured, 12% sodium dodecyl sulfate-polyacrylamide (SDS-PA) They were loaded into gels. The separation process was carried out at 75 V for approximately 3 hours. The gel preparation and processing procedures were carried out using the Mini-PROTEAN Tetra Handheld Gel. It is manufactured using a casting system and a PowerPac™ Base Power Supply (BioRad). For comparative analysis of protein expression levels. For normalization purposes, β-actin protein was used as a reference. This method is effective for both... SLKUR and free curcumin have effects on the expression of apoptotic proteins 10 It presents this in detail. In a prostate cancer model, 22Rv1 xenograft Treatment efficacy was evaluated using the (Foxn1nu / Foxn1nu) model. Nude male mice were preferred. During model development, 12x10⁶ 22Rv1 cells were used in a 1:3 ratio. It is dissolved in Matrigel at a certain ratio and injected subcutaneously into the dorsal region of mice. It has been done. 15 In vivo experiments with soy lecithin-based encapsulated curcumin showed no control results. (Sterile PBS), soy lecithin (SL), free curcumin (KUR) and soy lecithin-based encapsulated Homozygous nude (Foxn1nu / Foxn1nu) males treated with curcumin (SLKUR). Tumors were isolated from mice (n=5). The volume of tumors collected from each animal was 20 The tumor volume was calculated using the formula “Tumor volume = 1 / 2(Length × (Width)2)” (Table 1). According to the results, the mean tumor volume in the animal group treated with SLKUR was greater than in the control group. It is 1 / 10 smaller than the group (SLKUR=143 mm3, control= 1032 mm3, (KUR=723 mm3, SL=584 mm3). Thus, the anti-tumor effect of SLKUR is greater than that of KUR. It has been determined that there are more than 25. On day 4 following cell implantation, animals were randomly divided into 4 groups: 1. Control Group: Group treated with sterile dH₂O (n=6), 2. Soy Lecithin (SL) Group: Soy lecithin administered intraperitoneally at a dose of 1.5 g / kg. The group that was treated (n=6), 30 3. Free Curcumin (CUR) Group: Administered intraperitoneally at a dose of 500 mg / kg. Group treated with free curcumin (n=6), 4. Soy Lecithin-Curcumin (SLKUR) Group: Administered intraperitoneally at a dose of 1.5 g / kg. Group treated with curcumin encapsulated with soy lecithin (n=6). 17 The first injection is given on the 4th day after the implantation of cancer cells. The procedure was performed and then injections were given every 4 days for 3 weeks. From the time of cell implantation, the animals were weighed every 4 days and Tumor volume was measured weekly. Tumor volume was calculated according to Formula 1. Formula 1. 𝑇ü𝑚ö𝑟 𝐻𝑎𝑐𝑚𝑖 0.5 𝑈𝑧𝑢𝑛𝑙𝑢𝑘 𝑥 𝐺𝑒𝑛𝑖ş𝑙𝑖𝑘 In the fourth week of the study, the animals were sacrificed and tumor tissues were pathologically examined. They were collected for examination. Histopathological analysis of the tumors was performed using the Gleason score method. An evaluation was conducted. In addition, 10 isolated samples were taken from all animals in the experimental groups. organs that are removed (brain, heart, lungs, liver, pancreas, stomach, kidneys, spleen, testes) The presence of toxic effects on the testicles and genitals has been investigated. Within this scope, the organ... Any signs of toxicity in the tissues were examined in detail. The subject of the invention... In vivo experiments were conducted to evaluate the anti-tumor efficacy of SLKUR. As a result, the control group (Sterile PBS) contained soy lecithin (SL), free curcumin (KUR), and 15 homozygous nude treated with soy lecithin-based encapsulated curcumin (SLKUR) Analysis of results from tumors isolated from (Foxn1nu / Foxn1nu) male mice (n=5) The volumes of tumors collected from each animal were determined using Formula 1. calculated (Faustino-Rocha et al., 2013). The results are given in Table 1. Table 1. Tumor volumes collected from each animal group. Control (mm3) SL (mm3) KUR (mm3) SLKUR (mm3) 3528 1590 1589 108 686 484 147 125 1013 324 845 63 350 400 1014 224 930 123 24 196 1032 584 723 143 (SL: Soy lecithin, KUR: Curcumin, SLKUR: Soy lecithin-based encapsulated curcumin) 18 According to the experimental results, the average tumor volume in the group treated with SLKUR was... It was found to be approximately 1 / 10 lower compared to the control group. Groups The average tumor volumes between them are as follows:  Control Group: 1032 mm³  Soy Lecithin Group: 584 mm³ 5  Free Curcumin Group: 723 mm³  Soy Lecithin-Based Encapsulated Curcumin Group: 143 mm³ These results show that SLKUR has a higher anti-tumor effect than KUR. This is demonstrated by the significant volume reduction, particularly in the SLKUR group. This shows that the formulation is superior in terms of targeting and efficacy (Figure 7). The studies conducted within the scope of the invention include the findings shown in Figures 3, 4, 5 and 6. All data analyses in the graphs were performed using the GraphPad program, and The results are graphed in this program. The data can be presented in a parametric or non-parametric format. After determining whether it has a distribution, data exhibiting a parametric distribution The analysis was performed using the Student-T test in comparison with the control group. Non-parametric. For data showing a distribution, Analysis of Variance (ANOVA) test was applied. Each experiment was repeated at least 5 times. A p-value <0.05 was considered statistically significant. The significance levels are indicated as follows: 20  *P ≤0.05  **P ≤0.01  ***P ≤0.001  ****P ≤0.0001 30 19 REFERENCES [1] Diomede, F.; Fonticoli, L.; Guarnieri, S.; Della Rocca, Y.; Rajan, T.S.; Fontana, A.; Trubiani, O.; Marconi, G.D.; Pizzicannella, J. The Effect of Liposomal Curcumin as an Anti-Inflammatory Strategy on Lipopolysaccharide from Porphyromonas gingivalis Treated Endothelial Committed Neural Crest Derived Stem Cells: Morphological and 5 Molecular Mechanisms. Int. J. Mol. Sci. 2021, 22, 7534. [2] Bala, P., et al. (2021). Controlled release of curcumin using polymer-based carriers and anti-cancer efficacy. Journal of Biomedical Research, 35(2), 150-160. [3] Sharma, R., et al. (2020). Enhancement of curcumin bioavailability using chitosan nanoparticles. International Journal of Pharmaceutical Sciences, 42(3), 250-260. 10 [4] Mukherjee, S., et al. (2019). Curcumin delivery using solid lipid nanoparticles and anti-cancer effects. Journal of Drug Delivery Science and Technology, 31, 20-30. [5] Wang, Y., et al. (2022). Controlled release of curcumin using polymeric microspheres. European Journal of Pharmaceutics and Biopharmaceutics, 40(1), 75- 85. 15 [6] Chen, L., et al. (2021). Hydrogel-based curcumin delivery systems. Journal of Advanced Materials, 36(5), 305-315. [7] Ranjan, A., et al. (2021). Preparation of curcumin-loaded nanofibers using electrospinning and their effects. Journal of Nanoscience and Nanotechnology, 34(4), 180-190. 20 30

Claims

REQUESTS 1. Curcumin encapsulated for use in the treatment of prostate cancer. Its distinguishing feature is that it is soy lecithin-based.

2. 5 encapsulated tablets according to Claim 1 for use in the treatment of prostate cancer. It is curcumin, and its characteristic feature is that the aromatic part C=C is present at 1627 cm-1 in the FTIR spectrum. stress, benzene ring stress vibration at 1589 cm−1 and C=O at 1516 cm−1 and C=C means it includes vibration peaks.

3. Encapsulated according to Claim 1 for use in the treatment of prostate cancer. It is a method of synthesizing curcumin and its characteristic is; 10 i. For the preparation of soy lecithin dispersion, sterile soy lecithin is used. Mixing with injection water in a homogenizer, ii. In order to reduce the particle size of the resulting dispersion, type Performing the procedure using a sonicator device, iii. The dispersion is freeze-dried and powdered using a lyophilizer. 15 bringing, iv. the powder is left to stand in a vacuum oven and then tightly sealed. Soy lecithin should be stored in containers in the refrigerator. obtaining nanoparticles, v. Prepared soy lecithin nanoparticles and pure curcumin in sterile 20 mixing with water for injection, vi. The mixing process is carried out using a type sonicator. vii. the resulting soy lecithin-based encapsulated curcumin dispersion Drying after freezing using a lyophilizer, viii. then, mixed once more with a sonicator under the same processing conditions 25 processing, ix. Re-frozen soy lecithin-based encapsulated curcumin re-drying of the dispersion with a lyophilizer, x. The resulting soy lecithin-based encapsulated curcumin was placed in a vacuum oven. After being left to stand, store in tightly sealed containers in the refrigerator for 30 days. hiding It includes the steps of the process. 21 4. Encapsulated according to Claim 1 for use in the treatment of prostate cancer. This is the method of synthesizing curcumin and its characteristic feature is; i. To prepare a 2% (weight / volume) dispersion of soy lecithin. Soy lecithin was injected with sterile water for 15-30 minutes at 21,000 rpm. mixing in a homogenizer with a value of 5 ii. In order to reduce the particle size of the resulting dispersion, type Using a sonicator device, 5 each at 70 kHz power and 70% vibration. The process takes place in three 15-minute intervals, for a total of 15 minutes. to be done, iii. The dispersion was lyophilized at -50 °C and 0.1 millibar 10°C using a lyophilizer. freeze-drying under pressure and grinding into powder, iv. The powder is left in a vacuum oven for 1 hour and then tightly sealed. Soy lecithin should be stored in sealed containers in the refrigerator. obtaining nanoparticles, v. Prepared soy lecithin nanoparticles and pure curcumin, in a 3:1 ratio 15 weighed and mixed with sterile water for injection, vi. Mixing process using a type sonicator at 70 kHz power and 70% in vibration mode, for a total of 45 minutes at 5-minute intervals. to be carried out, vii. 20 of the obtained soy lecithin-based encapsulated curcumin dispersion After freezing, it is processed using a lyophilizer at a temperature of -50 °C and 0.1 drying under millibar pressure, viii. then, once again for 45 minutes under the same operating conditions (5 (at minute intervals, with a sonicator at 70 kHz power and 70% vibration) mixed and processed, 25 ix. Re-frozen soy lecithin-based encapsulated curcumin dispersion of the mixture in a lyophilizer at -50 °C and 0.1 millibar pressure drying again underneath, x. The resulting soy lecithin-based encapsulated curcumin was placed in a vacuum oven for 1 hour. After being left to stand for a certain period, store in tightly sealed containers in the refrigerator for 30 days. hiding It includes the steps of the process.

5. For use in the treatment of prostate cancer, according to either Order 3 or 4. Curcumin encapsulated using soy lecithin synthesized by a specific method.

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