INTRODUCTION:

Breast cancer is a leading cause of female mortality and poses a significant risk to women's lives. In developing nations, challenges such as limited resources and inadequate infrastructure hinder efforts to improve breast cancer prevention through timely check-ups, testing, and control.^1,2 Cancer can be cured if detected and treated promptly. Various treatments, such as surgery, radiation therapy, chemotherapy, immunotherapy, and hormonal therapy, are available, though each has its drawbacks.³ Chemotherapy drugs may harm healthy cells along with cancerous ones, emphasizing the need for an alternative method that is both effective and safe for treating cancer.^4,5 Certain plants with cancer-fighting properties are increasingly being used as natural alternatives to traditional cancer treatments, contributing to the growing popularity of herbal remedies. Ellagic acid and coumaric acid are natural polyphenols in the subclass of phenolic acids.⁶ The structure of ellagic acid resembles a dimeric benzoic acid derivative with four OH groups and two lactone rings, indicating its hydrophilic nature. It is naturally found in fruits and nuts like raspberries, strawberries, blueberries, pomegranates, and grapes, either as glycoside derivatives or ellagitannins. Research explores the potential benefits of ellagic acid, such as its antiproliferative properties against cancer, its ability to kill cancer cells, and its role in inhibiting angiogenesis and metastasis.⁷ Coumaric acid is chemically 4-hydroxy-cinnamic acid, is a vital anti-allergic compound found in grains, fruits, and vegetables. Studies on various tumor cell lines, such as those from hepatocellular cancer, lingual epithelium, and colorectal carcinoma, have shown that p-CoA inhibits their proliferation and spread.^8,9 However, the limited solubility and bioavailability of ellagic acid and coumaric acid hinder their full clinical use and medical benefits. This inadequate bioavailability is primarily due to poor absorption, metabolism in the GIT, the first-pass effect, and rapid excretion when administered orally. These problems can be successfully resolved and medication can be precisely administered with the use of nanotechnology.¹⁰

The research work involves encapsulation of polyphenols within natural polymers to develop the polyphenol nanoparticles. Polymeric encapsulation involves shielding, capturing, binding, or regulating the gradual release of medicinal moieties within their matrix.^11,12 By combining ellagic acid and p-coumaric acid within a nanoparticle formulation, we anticipate potential synergistic effects, resulting in enhanced efficacy compared to using them individually. Nanotechnology can encapsulate ellagic and coumaric acids in biodegradable polymeric nanoparticles.¹³ Polymeric nanoparticles have unique properties that can benefit human health by protecting, encapsulating, targeting, or releasing therapeutic substances for cancer treatment. This approach holds promise for targeting various pathways implicated in breast cancer development and progression. Sodium alginate, a biodegradable polymer, is used to encapsulate the drug. Glyceryl mono oleate (GMO) is used as lipid phase and with its sticky appearance, targets carcinoma cells. Howevr, poloxamer 188, a copolymer with anchoring characteristics is used to enhance the dissolution of hydrophobic substances. Besides, developement of novel dosage forms necessiates evidences for anti-cancerous therapeutic efficacy through its moderation and effective modulated targeted delivery systems.¹⁴

The present study represents development of polyphenolic nanoparticles as novel drug delivery systems being an advanced alteration from traditional polyphenol formulations. These nanoparticles offer several benefits, including improved bio-distribution, heightened therapeutic efficiency, and decreased nonspecific toxicity of potent anticancer agents. They effectively enhance solubility, stability, biocompatibility, and protection from degradation. Neverthless, these nanoparticles facilitate the targeted delivery of therapeutic agents to cancer cells in laboratory settings, leading to reduced dosages and improved pharmacological activity to fulfill patient’s unmet needs.¹⁵

MATERIAL AND METHODS:

Materials:

Poloxamer 188 (P188) was obtained from Research Lab. Chem. Mumbai, Glyceryl monooleate (GMO) received from Mohini Organics Mumbai, Sodium alginate and Dimethyl sulfoxide procured from Loba Chem. Pvt. Ltd. Mumbai and; Ellagic acid and Coumaric acid was gifted by Otto Biochemika Mumbai.

Methods:

Formulation of Sodium Alginate Polyphenolic Nanoparticles:

Sodium alginate nanoparticles were preapred by precisely weighing 10mg of ellagic acid and 10mg of p-coumaric acid. These nanoparticles were added to 100mL beaker to dissolve in 200mg of molten GMO. Furthermore, 6.5mL of P188 in varying concentrations of 0.5%, 1%, and 1.5% was added to the previously prepared mixture and sonicated for 10min using probe sonicator to produce an o/w emulsion. Once more, 6.5mL of the sodium alginate solution of different concentrations 1.2%, 2.4%, and 3.6% was dropwise added to the aforementioned emulsion and sonicated for 10min using probe sonicator. This phase was subjected to 40 cycles of high-pressure homogenization at 800 psi pressure to develop a nanoemulsion, which was freeze-dried over 48h using 2% w/v mannitol as a cryoprotectant.¹⁶

Development of Sodium Alginate Polyphenolic Nanoparticles:

A central composite design (CCD) with combined effects of two variables namely; A: P188 (0.5, 1% and 1.5 %) and B: sodium alginate (1.2%, 2.4% and 3.6%), each at three levels was explored to examine nine different combinations. A full factorial design employed for optimizing the formulation of polyphenolic nanoparticles with varying two independent varaibles at three levels each. The goal was to find the best proportion of polymer in combination yielding improved formulation characteristics. Factorial design aligns with quality by design and one factor at a time principle by systematically studying the factors influencing the formulation. Design of experiment (DOE) is a structured approach to experimentation that aims to optimize processes and understand their underlying relationships.¹⁷

Table 1: Composition of polyphenolic nanoparticles

Batch No.	Drug (mg)		GMO (ml)	P188 (%)	Sodium alginate (%)
Batch No.	Ellagic acid	p- coumaric acid	GMO (ml)	P188 (%)	Sodium alginate (%)
1	10	10	2	0.5	1.2
2	10	10	2	1	1.2
3	10	10	2	1.5	1.2
4	10	10	2	0.5	2.4
5	10	10	2	1	2.4
6	10	10	2	1.5	2.4
7	10	10	2	0.5	3.6
8	10	10	2	1	3.6
9	10	10	2	1.5	3.6

Evaluation of the Sodium Alginate Polyphenolic Nanoparticles:

a) Estimation of percent entrapment efficacy and percent loading efficiency:

Accurately weighed 20mg of polyphenolic nanoparticles were dissolved in 10mL of dimethyl sulfoxide (DMSO) and the acquired residue was rinsed and diluted with lightly shaking for 24h at 37 °C. The supernatant was collected after centrifugating mixture at 16,000g for 15min. Accurately, 1mL fraction of supernatant was diluted to 10mL with DMSO to liberate entrapped nanoparticles and the absorbance was measured using UV spectrophotometer at 254nm for ellagic acid and 285nm for p-coumaric acid. The percent entrapment efficacy (%EE) and percent loading efficacy (%LE) was calculated using eq. (1) and (2).¹⁸

Amount of drug incorporated in sodium alginate nanoparticles

% EE= --------------------------------------------------------- X 100

Total amount of drug added

(1)

Amount of drug present in sodium alginate nanoparticles

% LE= ---------------------------------------------------------- X 100

Total weight of nanoparticles (2)

b) Particle size and zeta potential analysis: Polyphenolic nanoparticles were tested for particle size analysis. The average size of the particles was measured using dynamic light scattering with the HORIBA SZ 100 zeta sizer. To ensure accurate results, the nanoparticle solution was diluted with phosphate buffer saline to eliminate multi-scattering. Analysis was carried thrice to achieve accuaracy and efficiecny of method. The sample of polyphenolic nanoparticle was diluted ten times before measurement. The disposable capillary cell was used, and the analysis was done three times for accuracy. Zeta potential indicates the surface charge of particles and can affect their stability and interactions.¹⁹

Characterization of Sodium Alginate Polyphenolic Nanoparticles:

a) Fourier Transform Infrared Analysis: The phytoconstituents were identified by analyzing its unique infrared absorption bands. Study represents phytoconstituent’s potential interactions with excipients by detecting changes in the FTIR spectrum. It help to assess the compatibility of polyphenol-loaded solid lipid nanoparticles by comparing the FTIR spectra of the unadulterated phytoconstituent and the optimized SLN formulations in the range of 4000 to 400 cm^-1.²⁰

b) X-ray Diffraction Analysis: X-ray diffractometer D2 Phaser, Bruker AXS instrument was empoyed to examine a phytoconstituents and its polyphenol-nanoparticles. XRD helps to determine the existance of crystallinity or amorphisation within the material. The analysis involved exposing the sample to X-rays (copper Kα radiation) at a specific energy (40kV and 45mA). The scan covered an angle range (2θ) from 5 to 90°, with the angle changing at a speed of 2θ per min. The resulting XRD patterns were studied to indentify changes in the peak intensity which indicates shifts between crystalline and amorphous forms.²¹

c) Differential Scanning Calorimetry (DSC) Analysis: DSC was used to find out the critical information of the phytoconstituents and excipient such as purity, stability, compatibility. Thus, investigated the thermal stability of compound (SDT Q600 V20.9 Build 20). Approximately 3-5mg of lyophilized sample was crimped in an aluminium pan with a heating rate (20-680˚c) at 10˚C min^-1.²²

d) Transmission Electron Microscopy (TEM) Analysis: The TEM examination was conducted by diluting the sample of polyphenolic nanoparticle and applying it to a carbon-coated grid. The grid was negatively stained with 1% phosphotungstic acid. The stained sample on the grid was allowed to air dry. The dried sample was examined using TEM at an accelerating voltage of 80kV at 25000. This process helps visualize the structure and features of the vesicles at a very high magnification.²³

In Vitro Dissolution Study:

In vitro drug release of polyphenolic nanoparticles was worked out to examine the drug release profile. The procedure included immersion of sodium alginate nanoparticles into phosphate buffer saline (PBS) solution of pH 7.4 using dialysis membrane sac over 24 h. A dialysis bag carrying medication was submerged into a beaker consisting 50mL of PBS solution. The container was placed into incubator programmed to shake continuously at 37°C (80 –100rpm). Accurately, 5 mL of the supernatant was withdrawn at definite time intervals, and examined using UV spectrophotometer to estimate the quantities of ellagic acid and p-coumaric acid, at wavelength 254 and 285nm, respectively.²⁴

Stability Study:

Stability study was carried out to examine the impact of environmental parameters like temperatures, light on the prepared herbal nanoparticles. According to ICH guidelines 3 months stability study was performed on herbal nanoparticle.²⁵

In Vitro Cytotoxicity by MTT Assay:

a) Cell Culture: In vitro cytotoxicity study of sodium alginate polyphenolic nanoparticles evaluated using MCF-7 human breast cancer cell line, were seeded in 96-well culture plates (1 × 10⁴ cells/well) and cell culture media DMEM medium supplemented with 10% Foetal Bovine Serum (FBS), MP Biomedicals, Germany, incubated at 37^oC and 5% CO₂atmosphere for 24h.²⁶

b) Cell Viability Assay: Each well of the 96-well microtiter plate received 200µl of the cell suspension, which was subsequently incubated for 24h at 37°C with 5% CO₂. After 24h, the utilized media was aspirated. The relevant wells received 200µL of varied test drug concentrations. Further, the plate was incubated for 24h at 37°C and 5% CO₂. The drug consisting medium was aspirated when the plate was taken out of the incubator. Accurately, 200µL medium containing 10% MTT reagent were added to each well to obtain a final concentration of 0.5mg/mL. The plate was then incubated for 3h at 37°C and 5% CO₂. Without disrupting the crystals already produced, the culture media was totally removed. To dissolve the produced formazan, 100µL of solubilizing solution (DMSO) was added, and plate was carefully shaken using gyratory shaker. Both, a 570nm and a 630 nm, wavelengths were used to measure the absorbance on a microplate reader. The quantity of test medicine required to inhibition cell growth by 50% (IC₅₀) was calculated using the cell line's dose-response curve after the background and the blank had been eliminated.²⁷

RESULTS AND DISCUSSION:

Formulation of Sodium Alginate Nanoparticles:

The optimal formulation identified consists of 2.4% sodium alginate and 0.5% poloxamer (188). Under these conditions, the nanoparticles achieved a particle size of 163.91nm and a zeta potential of -25.5mV, which indicates good stability. It represents that a combined effect of electrostatic and sterical stabilization a zeta potential of about -20mV can be sufficient for physical stability. Hence, formulation in investigation have shown prospects for physical stability as it has zeta potential of -25.5±1.17mV. Values of zeta potential showed that prepared polyphenolic sodium alginate based nanoparticles have sufficient charge and mobility to inhibit aggregation of particles. The negative zeta potential suggests that the particles are well-dispersed and less likely to aggregate, which is essential for the effective delivery of anticancer agents. The desirability index of 1.000 signifies that the optimized formulation meets all the desired criteria, including particle size, zeta potential, and overall stability, making it the most favorable option for further development in cancer treatment. Polyphenolic nanoparticles prepared by probe sonication method and high-pressure homogenizer by utilising GMO as lipid phase, sodium alginate as polymer, P188 as copolymer, ellagic and p- coumaric acid as active pharmaceutical ingradients suffices the counter ions for neutralization within electriacally double diffused layer and hinders stability.

Design of Experimention (DOE):

a) Effect of independent variables on particle size (Y₁):

The relationship between particle size in the polyphenolic nanoparticles, P188, and sodium alginate concentrations is illustrated through Table 2. The K4 formulation demonstrated the most favourable particle size. To capture the influence of the independent factors on particle size, a quadratic equation can be employed. This equation helps depict the impact of various factors on particle size within the context of study.

Particle size (Y₁)

= +142.67 + 5.50X₁ - 20.80X₂ (3)

Where, Y₁ is the Particle size, X₁ is the concentration of sodium alginate and X₂ is the concentration of P188. From the equation, it is clear that P188 has a negative effect on the particle size and the concentration of sodium alginate has a possitive effect. This means that the particle size of formulation decreases with increase in concentration of P188 increases and increases with an increase in the concentration of sodium alginate.^28,29

Table 2: Development of nanoparticle batches by applying DOE

Formulation Code	Independent parameters				Dependent parameters
Formulation Code	Std	Run	^X₁ Sodium alginate (%)	^X₂ P 188 (%)	^Y₁ Particle size (nm)	^Y₂ Zeta potential (mV)
K1	5	1	1.2	0.5	159.4	-18.5
K2	7	2	1.2	1	126.7	-31.2
K3	1	3	1.2	1.5	127.6	-43.5
K4	4	4	2.4	0.5	163.1	-24.3
K5	2	5	2.4	1	131.7	-36.9
K6	3	6	2.4	1.5	128.8	-46.2
K7	6	7	3.6	0.5	179.6	-30.8
K8	9	8	3.6	1	146.2	-38.9
K9	8	9	3.6	1.5	120.9	-49.8

The Model F- value of 14.75 implies the model is significant. There is only a 0.48 % chance that an F- value this large could occur due to noise p-values less than 0.0500 indicate model terms are significant. In this case B is a significant model term. Values greater than 0.1000 indicates the model terms are not significant. If there are many insignificant model terms (not counting those required to support hierarchy), model reduction may improve your model. The plot indicates a strong correlation between the predicted (fitted) response values generated by the regression model and the actual observed values. The middle line represents an unbiased 1:1 relationship between predicted and actual values. When the points on the graph are close, with no clear pattern in residuals and uncertainties are within limits, the predictions are accurate. Outliers, where points are significantly distant, suggest issues with the data. If all points are on the predicted side, it indicates overestimation of predicted values compared to actual values. Scatter plot is showing predicted values against real observed values with a fitted regression line through the points, with most of them are close to the line.

ANOVA table explaining data variability and model impact with reference to full factorial design. Response surface plot includes a diagonal reference line for perfect prediction alignment with label axes, title, and ensuring clarity in the presentation. It depicts the impact on one of the dependent variable particle size, study showed that increasing P188 concentration led to smaller polyphenolic nanoparticle sizes, while sodium alginate concentration resulted in negative correlation with reference to the same dependent variable.³⁰ These findings were represented using 3D- response graphs, visually depicting the impact of independent variables on particle size.

a) Effect of independent variables on zeta potential (Y₂): Study displyed zeta potential is linked to the concentration of P188 and sodium alginate of the formulation. The results of zeta potential of the polyphenolic nano formulations are shown in Table 2. The effect of independent variables on the zeta potential can be explained by the following quadratic equation:

Zeta potential (Y₂)

= -35.56X₁ - 4.37X₂ - 10.98X₁X₂+ 1.50 (4)

Where Y₂ is the dependent variable zeta potential, X₁ is the concentration of sodium alginate and X₂ is the concentration of polymer P188. From the equation, it is clear that P188 has negative correlation with reference to a zeta potential and the concentration of sodium alginate also has a negative effect. This means that the zeta potential of polyphenolic formulation decreases with an increase in the concentration of P188 as well as sodium alginate. Concentration of indepenedent variables are optimized in such a way that it exhibits suffcient stabilization to the diffused layer and avoids aggreration of nanoaprticles from delivery systems.³¹

The Model F- value of 416.14 implies the model is significant. There is only a 0.01% chance that an F- value could occur due to signal to noise ratio. p-values less than 0.0500 indicate model terms are significant. In this case A, B, AB are significant model terms. Values found to be greater than 0.1000 indicates the model terms are not significant. If there are many insignificant model terms, model reduction may improve your model. The three dimestional graphs show that increasing P188 concentration from 0.5% to 1.5% reduces nanoparticles zeta potential. This suggests that both P188 and sodium alginate concentrations affect surface charged particles from nanoformulations.³² Overlay graph for optimized Batch K4 depicting the design space for selecting optimal concentrations of P188 and sodium alginate. It shows that the optimized effective concentration of P188 is 0.5%, while for sodium alginate, it’s 2.4%. These optimal values align very closely with the model's design points space.

Analysis of percent entrapment efficiency and percent loading efficiency: Entrapment efficiencies play a vital role in the drug's initial immediate release lag followed by sustained release of actives. The respective formulation comprising sodium alginate and P188 based nanoparticles has entrapment efficiencies of 77.38±1.34% and 82.87±0.46%, with loading capacities of 4.07±1.46% and 4.25±1.06% for ellagic acid and coumaric acid, respectively.

Particle size and zeta potential analysis: The particle size of all the polyphenolic nanoparticles batches found in range between 120.9 nm to 179.6 nm. The nanoparticle size of optimized batch K4 was found to be 163.1 nm. (Figure 1 A) The zeta potential of all the polyphenolic batch K1-K9 which was found within the range -18.5 mV to -49.8mV. The zeta potential of optimized batch was found to be -24.3 mV. (Figure 1 B).

Characterization of Sodium Alginate Polyphenolic Nanoparticles

a) FTIR Study: In the spectrum of ellagic acid (Fig. 2A) there is a broad O-H stretching from hydroxyl groups appears at 2800–3700 cm⁻¹, often strong due to hydrogen bonding. C=O stretching from lactone rings shows around 1725 cm⁻¹. Coumaric acid (Figure 2B) broad O-H stretching from the hydroxyl and carboxylic groups appears at 2500–3500 cm⁻¹ due to hydrogen bonding.³³ C=O stretching from the carboxylic acid shows a strong peak around 1680–1720 cm⁻¹. The C=O functionality of GMO (Fig. 2C) was seen with a strong peak at 1738 cm^-1. P188 (Fig. 2D) showed a strong C-H stretching from alkyl chains appears at 2800–3000 cm⁻¹, with C-O-C ether stretches prominent at 1100–1200 cm⁻¹. C-O and C-H deformations show around 1350–1450 cm⁻¹ and sodium alginate (Fig. 2E) broad O-H stretching from hydroxyls appears at 3200–3600 cm⁻¹. Asymmetric and symmetric COO⁻ stretches showed at 1600–1650 cm⁻¹ and 1400–1450 cm⁻¹, respectively. Sodium alginate nanoparticles (Figure 2F) displayed OH stretching peak at 3191.60 cm⁻¹ and C=C stretching peaks at 1451.34, 1360.01, 1332.99, 1255.99, and 1194.81 cm⁻¹. The C-H stretching vibrations occur at 928.12 cm⁻¹ for carboxylic groups and at 707.94 and 625.68 cm⁻¹ for aromatic groups. In nanoparticle spectra, these peaks may shift or broaden due to interactions with drugs/polymers like alginate, indicating adsorption without chemical bonding.^34,35

b) XRD Analysis: X-ray diffraction peaks showed for ellagic acid at 28.14° (Fig. 3A), coumaric acid at 17.62° (Fig. 3B), GMO at 19.36° (Fig. 3C), P188 at 19.06° (Fig. 3D), sodium alginate at 13.28° (Fig. 3E), and sodium alginate nanoparticles at 9.70° (Fig. 3F) characterize the XRD spectrum. The disappearance of the diffraction angle indicates the integration of biomolecules into the nanoparticle preparation in powdered amorphization state. This results in high encapsulation efficiency of the formulations of amorphous biomolecules.³⁶

Figure 3. XRD of polyphenolic nanoparticles (A) Ellagic acid, (B) Coumaric acid, (C) GMO, (D) P188, (E) Sodium alginate, and (F) Sodium alginate nanoparticles

c) DSC Analysis: A DSC thermogram of ellagic acid (Fig. 4A) exhibits a sharp endothermic peak at 469.37 °C, while coumaric acid (Fig. 4B) displays a peak at 222.21 °C, corresponding to their m.p. The GMO (Fig. 4C) shows an endothermic peak at 39.97 °C, P188 (Fig. 4D) showed an at 62.13 °C, and sodium alginate (Fig. 4E) has a broad peak around 93.65 °C. The sodium alginate nanoparticles (Fig. 4F) demonstrated endothermic peaks at 173.27 °C and 354.91 °C, occurring at lower temperatures than their pure forms and showing no interactions, just the superimposition of the specific elements. The representation of thermograms which indicate a reduction in their crystalline structure, signifying the complete encapsulation of coumaric acid and ellagic acid.^37,38

Figure 4. DSC endothermic peak of (A) Ellagic acid, (B) Coumaric acid, (C) GMO, (D) Poloxamer 188, (E) Sodium alginate, and (F) Sodium alginate nanoparticles

d) TEM Analysis: Nanoparticle morphology is typically examined using TEM due to its superior resolution compared to SEM. TEM reveals spherical nanoparticle shapes and shows darker patches within the spherical matrix of ellagic acid and poloxamer-loaded sodium alginate nanoparticles. (Fig. 5) The core-shell structure indicates that sodium alginate fully coats the individual particles.³⁹

Figure 5. TEM image of sodium alginate nanoparticles

In vitro drug release study:

As a result, sodium alginate nanoparticles showed significantly rapid drug releases of 75.83% and 81.63%, respectively for ellagic acid and coumaric acid at 24 h, Fig. 6. In vitro tests using the sodium alginate nanoformulation suggest a controlled release, indicating minimal sudden release due to either limited surface binding or strong attachment of ellagic acid and coumaric acid to the sodium alginate layer. Sodium Alginate has intrinsic property as rheology modifier by imparting viscosity to developed layers. Due to such changes in rheology, it leads to sustained release of drugs from novel formulations. Given the hydrophobic nature of these biomolecules, they are likely concentrated in the GMO core rather than on the hydrophilic sodium alginate surface. Successful extended and regulated release methods are achieved with sodium alginate for ellagic acid and coumaric acid.⁴⁰

Figure 6. In vitro drug release study

Stability Study:

Over a duration of two months, the stability study was carried out. The optimized batch K4’s particle size and zeta potential were calculated as a result of storage time. There was change in particle size or zeta potential after two months of observation.

In vitro cytotoxicity by MTT Assay:

The cytotoxicity of standard cisplatin (Fig. 7A), sodium alginate nanoparticles (Fig. 7B ) and untreated cell (Fig. 7C) studied on MCF-7 cell with their different concentrations ranging from 0.001 to 10μg/mL The IC₅₀ values for cisplatin and sodium alginate nanoparticles are found to be 8.7μg/mL and 10.48μg/mL, respectively. It is observed that the MCF-7 cell line remains stable even at low concentrations.

Figure 7. Image of cytotoxicity effect of (A) Standard drug, (B) Sodium alginate nanoparticles, (C) Untreated cell

DISCUSSION:

The study aims to develop sodium alginate polyphenolic nanoparticles with the smallest particle size while ensuring they remain stable and functional. Thus, the optimization process utilized in this study aims to fine-tune the formulation of sodium alginate nanoparticles to achieve the smallest possible particle size while maintaining their stability and functionality. Using a central composite factorial design, the researchers systematically varied the concentrations of sodium alginate and poloxamer 188 to identify the optimal conditions for nanoparticle preparation. The central composite design allows for the exploration of a broad range of formulation variables, providing valuable insights into how different parameters influence the final formulation. The overlay plot generated from this design helps to visualize the relationship between the formulation factors and the desired outcomes, enabling the researchers to select the most effective combination of ingredients. This approach is valuable for designing nanoparticles that can be precisely tailored to achieve desired properties, contributing to better drug delivery systems in biomedical applications. Targeted drug delivery is a critical area of research because it can reduce side effects and improve the effectiveness of treatments. By optimizing the nanoparticle formulation, the study aims to enhance the controlled release and bioavailability of drugs, especially in difficult-to-reach areas in the body, such as tumors or organs with specific barriers to drug penetration.This optimization significantly enhances the potential of the nanoparticles for targeted drug delivery, improving their therapeutic efficacy and stability for oral administration.

CONCLUSION:

This study was focused on development of a novel nanoparticle-based drug delivery system for the targeted treatment of breast cancer. The system uses sodium alginate nanoparticles to encapsulate ellagic acid and coumaric acid, which are water-insoluble compounds known for their anticancer properties. By utilizing sodium alginate with its inherent rheology modfication characteristics, the nanoparticles offer enhanced bioadhesion, controlled release with potentially improvement in the therapeutic efficacy of these bioactive compounds. In vitro tests, particularly the MTT assay, have been used to evaluate the cytotoxicity of these nanoparticles on breast cancer cells. The results suggest that ellagic acid and coumaric acid act synergistically increasing their anticancer potential. The study also highlights the advantage of oral administration as a potential route of delivery, as the sodium alginate platform appears to be effective in ensuring that the nanoparticles target cancer cells directly, reducing the likelihood of adverse effects on healthy tissues. This approach could help improve treatment outcomes for breast cancer patients by increasing the concentration of the therapeutic agents at the tumor site while minimizing systemic side effects. Overall, this work shows promise and leaves footrprints for advancing targeted cancer therapy using natural compounds in a biocompatible nanoparticle formulation. İt leads the scope for further studies in-vivo for long acting systemic circulation to achieve enhanced therapeutic efficacy of these biomolecules.

CONFLICT OF INTEREST:

Authors declare that there is no conflict of interest.

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Received on 28.12.2025 Revised on 22.03.2026

Accepted on 10.05.2026 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):2187-2195.

DOI: 10.52711/0974-360X.2026.00315

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