SYNERGISTIC ANTI-TUMOR EFFECTS OF NANO EMULSION FORMULATION CONTAINING CURCUMIN AND BRUCEA JAVANICA OIL: FORMULATION AND
CHARACTERIZATION
Chinnabathini Anilkumar
Asst. Professor, Department of Pharmaceutics, Princeton College of Pharmacy, Hyderabad, Telangana, India
Nandru Ramya
Asst. Professor, Department of Pharmaceutics, Princeton College of Pharmacy, Hyderabad, Telangana, India
Abstract -
Objective: The study's objective was to prepare and further enhance the antitumor effect of a nano emulsion (NE) infused with brucea javanica oil (BJO) and curcumin (CUR).
Methods and Materials: In a shaking incubator, the soluble levels of CUR and BJO in a variety of vehicles were measured. Pseudo-ternary phase diagrams were used to create NE in order to determine the component concentration range. Size, zeta potential, morphologies, physical stability, in vivo antitumor activity, and in vivo pharmacokinetics were used to investigate the NE's physicochemical and biological characteristics.
Conclusions: In optimized NE, the formulation of the BJO-CUR combination was successful. An anti-tumor activity experiment was used to determine whether BJO, which was used as the oil phase, had a synergistic antitumor effect with CUR. When compared to pure CUR, both the dissolution rate and oral bioavailability were enhanced.
Keywords: Curcumin, nano emulsion, pharmacokinetics, antitumor effect, and oil of Brucea javanica.
1 INTRODUCTION
The ripe fruits of Brucea javanica [L.]
Merr are the source of the complex mixture of fatty acids and fatty acid derivatives known as Brucea javanica oil (BJO). [Simaroubaceae It is accounted for that BJO has different diseases strong pharmacological exercises, for example, cancer suppressive, mitigating and antimalarial exercises. Additionally, it has effects on the clinical treatment of malignant tumors of intermediate and advanced stages. Bone marrow hematopoietic stem cell safety and improved quality of life for patients undergoing tumor treatment have both been demonstrated by BJO studies.
Curcumin, also known as CUR, is a naturally occurring, hydrophobic polyphenol that comes from the rhizome of the herb Curcuma longa. It includes anti-oxidant and anti-microbial properties in its extensive pharmacological profile [8, 9]. It also suppresses tumors in breast,
head, colon, and other cell lines. CUR is a promising drug for clinical treatment because of its low toxicity, as demonstrated by pharmacological studies.
However, the dissolution rate, metabolism, and permeability had been impacted by CUR's poor bioavailability and low solubility. The therapeutic effects and clinical application will then be limited by the low solubility. The creation of a formulation loaded with two therapeutic agents has become increasingly common, particularly in the treatment of cancer. In the field of chemotherapy for cancer, there are very few cases in which the disease is treated by a single agent. Both CUR and BJO have extensive anti-tumor spectra and favorable therapeutic effects, making them important players in cancer treatment. The therapeutic superiority of combination treatment over single drug treatment has been demonstrated by a
number of studies. To the best our insight, there is no writing written about the planning both of these two medications as NE.
The CUR and BJO's therapeutic effects have been limited by their low solubility and poor bioavailability. The significance of this article lies in selecting the appropriate drug delivery method. In order to sustain pharmacological action and achieve therapeutic local concentrations in a reasonable amount of time, the rate and extent of drug release from the carrier are crucial. The technology for nanoemulsification of drugs has advanced over the past ten years. The nanoscale droplet diameter, relative high rate of drug release from oil droplets into aqueous media, and high solubilization properties of nanoemulsion were identified as promising delivery systems.
It has received increasing attention for its beneficial properties because it is an efficient drug carrier that increases the absorption and solubility of poorly water-soluble drugs. Water, oil, surfactants, and co-surfactants made up nano emulsions. NE is a formulation that is thermodynamically stable. Depending on the oil phase, it is possible to avoid hepatic first pass metabolism and increase lymph directivity; Consequently, NE can also improve the skin's delivery of lipophilic and hydrophilic drugs. Because of its small droplet sizes, it prevented phase separation and improved membrane adhesion.
The formulation of a nanoemulsion requires a certain amount of oil. However, there were no reports regarding the BJO formulation as an oil for NE. The preparation of a nanoemulsion with BJO acting as the oil phase was the primary focus of this study. After that, a CUR-BJO-NE NE formulation with BJO and CUR was created. We wished mix utilizing of them could accomplish synergistic antitumor impact and improve oral bioavailability.
Characterization, a stability test, and an evaluation of the formulation followed.
Bioavailability. The portrayal of nanoemulsion was directed by it appearance of utilizing transmission electron microscopy (TEM) and drop size.
CUR-BJO-NE's oral bioavailability was studied in rats to compare its absorption to that of CUR suspension (CUR powder dispersed in 0.4% CMC-Na solution).
2 MATERIALS AND METHODS 2.1 Materials
The Sinopharm Chemical Reagent Co.
Ltd. in Shanghai, China, supplied the CUR. Yaoda Pharmaceutical Co. Ltd., based in Shenyang, China, supplied BJO.
Gattefosse (Shanghai, China) obtained Capryol 90. Aotai Chemical Ltd., based in Jinan, China, supplied the ethyl oleate.
Yihai Jiali Food Marketing Ltd. in Beijing, China provided the peanut oil. Polyoxyl 40 hydrogenated castor oil 40 (Cremophor RH 40) and Polyethylene glycol 400 (PEG400) was bought from BASF Co., Ltd.
(Germany) and Huadong substance reagent plant (Tianjin, China), individually. Meilin Industry and Trade Co., Ltd., based in Tianjin, China, supplied Tween 80. Fuchen chemical reagent factory in Tianjin, China, supplied the sodium carboxymethycellulose (CMC- Na). Analytical reagent grade was used for all other chemicals and solvents.
2.2 Animals
Vital River Laboratory Animal Center, located in Beijing, China, provided the male Wistar rats (250 20 g). The Laboratory Animal Centre of Hebei Medical University in Shijiazhuang, China, supplied the Kunming mice (weighing between 18 and 22 grams).
2.3 Cell lines
The Chinese Academy of Sciences' cell bank in Beijing, China, provided the mouse ascetic turnout cell line S180.
3 PREPARATION OF NES 3.1 Solubility study
The following measurements were made and carried out on the solubility of CUR and BJO in various vehicles: First, a quantity of BJO was added to each centrifugal tube with 1 mL of selected vehicles. The mixture was then kept in centrifugal tubes for two days at 25°C in a shaking incubator to reach equilibrium.
The degree of uniformity in mixing quality was chosen as the vehicle indicator. In addition, excess CUR was separately added to 1 mL of various oils and surfactants in the centrifugal tube, mixed for 48 hours in a shaking incubator at 25°C, and the solution was centrifuged at 10000 rpm for 10 minutes to remove the excess CUR. The UV spectrophotometer measured the concentration of CUR in the supernatant after the appropriate dilution with ethanol.
3.2 Construction of Pseudo-ternary Phase Diagrams
To determine the component concentration range for the current NE region, pseudo-ternary phase diagrams were constructed. Distilled water was added drop by drop to the mixture under proper magnetic stirring at room temperature until the mixture became clear at a certain point. Different weight ratios of oil phase, surfactants, and co- surfactants were mixed together.
3.3 Pharmacokinetics study of CUR- BJO-NE in rats
Two groups of five Wistar rats were randomly divided into. The Institutional Animal Care and Use Committee gave its approval to this animal experiment, which followed all of the regulations. The rodents were abstained for the time being before explore different avenues regarding free admittance to water. CUR-BJO-NE (400 mg/kg of body weight) and CUR suspension (1000 mg/kg of body weight, pure CUR powder dispersed in 0.4%
CMC-Na solution) were administered
intragastically to rats. At predetermined intervals, 0.5 milliliters of blood were taken from the ophthalmic vein of rats and centrifuged for ten minutes at 4000 rpm. The blood samples were then kept at -20 degrees Celsius until the analysis. 0.2 ml plasma was added with 0.6 ml ethanol and it was for 5 min vortex. The mixture was then centrifuged for ten minutes at 4000 rpm. HPLC analysis was used to determine the supernatant's CUR concentration.
3.4 In Vivo Antitumor Activity
The CUR-NE in vivo antitumor activities were contrasted with the established CUR-BJO-NE against mouse model bearing the S180 cell line. The following was the structure of the experiments:
Ascites formed in about seven days after tumor-bearing mice were inoculated intraperitoneally with mouse sarcoma cells from the S180 cell line. Then, at that point, the ascites was removed and washed with PBS and acclimated to cell suspensions in the serum free medium at cell grouping of 2 × 107 cells/ml and 200 μL of the blend was embedded subcutaneously into the flank of bare mice to lay out cancer xenograft. At that point, the tumor mouse model was established. The mice were then evenly divided into four groups based on their respective weights. With ten mice per group, the treatment began.
3.5 Statistical Analysis
The data were presented as the mean minus the standard deviation for each of the study's three experiments. At p 0.05, a two-tailed unpaired Student's t-test was used.
3.6 Characterization of CUR-BJO-NE CUR-BJO-NE's optimized TEM morphology. NE had a uniform size and appeared to be spherical. The average size of the particles was about 51.5 nm.
Additionally, CUR-BJO-NE's physical stability was evaluated. The samples did
not change in appearance, homogeneity, or phase separation, breaking, or drug precipitation after centrifugation. After centrifugation, this demonstrated that the formulation remained physically stable.
4 DISCUSSION
The purpose of the solubility studies was to identify appropriate oils and surfactants in NE with a high drug- loading capacity for both CUR and BJO.
When choosing vehicles, other ingredients' emulsification properties were also taken into account. After that, PEG400 was chosen as the co-surfactant rather than Transcutol HP, despite the fact that Transcutol HP was more soluble than PEG400. As surfactants, Cremophor RH40 and Labrasol were able to achieve a greater emulsifying effect when combined than when used separately. This may be connected to a greater reduction in the oil–water interfacial tension. The combination of the following effects probably contributed to the increased bioavailability: First, CUR's dissolution rate was significantly increased in CUR- BJO-NE, and when the drug was kept in a dissolved state in NE, oral absorption could be increased; Second, the effect of P-glycoprotein on the drug efflux effect may also be diminished or inhibited by surfactant use; NE droplets were able to escape the reticuloendothelial system's uptake and phagocytosis due to the smaller droplet size, which also increased the drug's circulation time. The large surface area of NE and the use of surfactants in formulation may be the cause of the increased dissolution rate.
In comparison to the group of blank NE, the in vivo antitumor activity results demonstrated that CUR-BJO-NE and CUR-NE effectively inhibited tumor cell growth. It suggested that the S180 cell was protected from tumor growth by both CUR and BJO. Additionally, this investigated whether utilizing BJO as an oil phase to dissolve CUR had a synergistic anticancer effect. In
conclusion, the development of a potential CUR-BJO-NE formulation was strongly supported by all of the obtained results.
5 CONCLUSION
A NE system that successfully incorporated both BJO and CUR for oral administration was developed in this study. An anti-tumor activity experiment was used to determine whether BJO, which was used as the oil phase, had a synergistic antitumor effect with CUR.
During the centrifugation and water dilution experiment, the CUR-BJONE exhibited stability. When compared to pure CUR, the dissolution rate and oral bioavailability were significantly improved.
The current study proved that the combination of CUR and BJO could be used to treat cancer.
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