A nanocomposite drug delivery system

Abstract

The present disclosure provides a drug delivery system comprising trimetallic nanoparticles coated with biocompatible and water-soluble coating polymers, encapsulating the anticancer agent proscillaridin, for enhancing targeted tumor cell delivery while minimizing damage to healthy cells. More particularly to a drug delivery system containing tri-metallic nanoparticles composite loaded with an anticancer agent with polymeric coating with a potential to improve the antitumor reactivity and selectivity toward specific type of tumor cell lines.

Description

A NANOCOMPOSITE DRUG DELIVERY SYSTEMTECHNICAL FIELD

[0001] The present disclosure relates to drug delivery systems, and more particularly to a drug delivery system containing tri-metallic nanoparticles composite loaded with an anticancer agent with polymeric coating with a potential to improve the antitumor reactivity and selectivity toward specific type of tumor cell lines.BACKGROUND

[0002] Metal oxide nanoparticles have emerged as highly promising materials for cancer detection and therapy due to their unique properties. They possess a high payload capacity, magnetic field responsiveness, surface modification versatility to overcome biological barriers, and biocompatibility. Additionally, it has the ability to encapsulate and bind various therapeutics further enhances their pharmacokinetic properties.

[0003] A study by Md ABdus Subhan titled “Advances with metal oxide-based nanoparticles as MDR metastatic breast cancer therapeutics and diagnostics” examined the cytotoxic activity of single metal oxide nanoparticles such copper oxide (CuO), iron oxide (yFe2O3) and multi-metal oxide nanoparticles such as zinc, iron and copper oxide (CuZnFe2O3) in human breast cancer cells and highlighted the great potential of using it as an anticancer against human breast cancer (MCF-7) cells.

[0004] A study by Nomfundo Mahlangeni et. al titled “Biosynthesis of manganese oxide nanoparticles using Urginea sanguinea and their effects on cytotoxicity and antioxidant activity” discloses an investigation of the cytotoxic effects and antioxidant activity for manganese oxide nanoparticles (MnOx), wich was synithisized using Urginea sanguinea and its isolates, a homoisoflavone (urgineanin E) and a mixture of bufadienolides (proscillaridin A, scillaren A and 5a-4,5-dihydroscillaren A).

[0005] A study by Huiyun Zhang et. al titled “Redox -responsive PEGylated self-assembled prodrug-nanoparticles formed by single disulfide bond bridge periplocymarin-vitamin E conjugate for liver cancer chemotherapy” discloses a redox-responsive prodrug- nanoparticles (MPSSV-NPs) self-assembled by co-nanoprecipitation of PPM-vitamin E conjugate and a PEG derivative of linoleate (mPEG2000-LA) in water. The researchshowcases the potential of these MPSSV-NPs as a safe and multifunctional solution for utilizing cardiac glycosides in cancer treatment.

[0006] None of the prior art documents discloses a drug delivery system of anti-tumor proscillaridin that is loaded onto trimetallic nanoparticles composite and coated with coating polymers for targeting specific tumor cell lines.SUMMARY

[0007] Therefore, it is an object of the present disclosure to provide a drug delivery system that may include proscillaridin encapsulated in trimetallic nanoparticles composite and coated with coating polymers.

[0008] In aspects of the present disclosure, the trimetallic nanoparticles may include the formula MlxM2yFe2O4.

[0009] In aspects of the present disclosure, Ml and M2 may be selected from the group including Zn+2, Ni+2, Al+3, Mn+2, Ag+, Y+3, Nd+3, Ti+4, Cd+2, Dy+3, Gd+3, Cu+2, Yb+3, Eu+3, Zr+4, In+3, Cr+2, Pr+3, Sm+3, Ho+3, Er+3, Mg+2, La+3, and Ce+3

[0010] In aspects of the present disclosure, x and y may have numeric value between 0 and 1.[OH] In aspects of the present disclosure, the total sum of x and y may equal 1.

[0012] In aspects of the present disclosure, the coating polymers may include polyethylene glycol (“PEG”) and chitosan.BRIEF DESCRIPTION OF THE DRAWINGS

[0001] The disclosure will now be described with reference to the accompanying drawings, without however limiting the scope of the disclosure thereof, and in which:

[0002] FIG. la illustrates XRD pattern of Cuo.5Zno.5Fe204 nanoparticles sample.

[0003] FIG. lb illustrates XRD pattern of Cuo.5Zno.5Fe204 nanoparticles sample coated with chitosan (“CS”).

[0004] FIG. 1 c illustrates XRD pattern of Cuo.5Zno.5Fe204 nanoparticles sample coated with PEG.

[0005] FIG. 2 illustrates Scanning transmission electron microscopy (“SEM”) image for uncoated Cuo.5Zno.5Fe204.

[0006] FIG. 3 illustrates an SEM image for Cuo.5Zno.5Fe204 coated with PEG and CS.DETAILED DESCRIPTION

[0013] Embodiments of the present disclosure provide a drug delivery system that may include proscillaridin encapsulated in bimetallic nanoparticles composite and coated with coating polymers.

[0014] In embodiments of the disclosure, the bimetallic nanoparticles may include the formula MlxM2yFe2O4.

[0015] In embodiments of the disclosure, Ml and M2 may be selected from the group including Zn+2,Ni+2,Al+3,Mn+2, Ag+, Y+3, Nd+3, Ti+4, Cd+2, Dy+3, Gd+3, Cu+2, Yb+3, Eu+3, Zr+4, In+3, Cr+2, Pr+3, Sm+3, Ho+3, Er+3, Mg+2, La+3, and Ce+3

[0016] In embodiments of the disclosure, x and y may have numeric value between 0 and 1.

[0017] In embodiments of the disclosure, the sum of x and y may equal 1.

[0018] In embodiments of the disclosure, the coating polymers may include PEG and CS.

[0019] In embodiments of the disclosure, the encapsulating proscillaridin in trimetallic nanoparticles composite may enhance the antitumor reactivity toward specific types of tumor cell lines.

[0020] In embodiments of the disclosure, the trimetallic nanoparticles loaded with proscillaridin and coated with PEG and CS may demonstrate improved selectivity towards specific types of tumor cell lines, minimizing damage to healthy cells.

[0021] In embodiments of the disclosure, the coating polymers may enhance biocompatibility, cellular uptake, and targeted delivery of the therapeutic agent to specific cell types.

[0022] The disclosure will be further illustrated on the basis of examples and a detailed description from which further features and advantages may be taken. It is to be noted that the following explanations are presented for the purpose of illustrating and descriptiononly; they are not intended to be exhaustive or to limit the disclosure to the precise form disclosed.

[0023] In Examples, reference will be made to FIGS. 1-2.Example 1Synthesis of Copper-Zinc ferrite nanoparticles Cuo.5Zno.sFe204 and proscillaridin

[0024] In this example, tri-metallic ferrite nanoparticles were synthesized using an aqueous extract of Boswellia Carteri. In about 20 ml of distilled water, about 0.56 g of FeSCU 7H2O, about 0.24 g of Zn(CH3CO2)22H2O, and about 0.24g of Q1SO45H2O were dispersed under stirring at room temperature for about 1 hour, resulting in a homogeneous solution. Then, about 25 ml of the aqueous plant extract was added dropwise, under continuous stirring for another about hour while controlling the pH to reach a range of 10-12. The solution was centrifuged, then the solid products were collected by magnetic filtration, washed several times with distilled water and ethanol, and finally dried in a vacuum oven at about 100 °C for about 6 hours.Coating of CuO.5ZnO.5Fe2Q4 nanoparticles and proscillaridin

[0025] In this process, Cuo.5Zno.5Fe204 nanoparticles and about 4pL of proscillaridin extract were added to theCS solution (10% by weight of CWO.JZWO.JF^CC nanoparticles of chitosan solution). Stirring continued until an opalescent hazy dispersion was visually observed, indicating the formation of the nanoparticles. Then PEG and CS solutions were prepared separately and mixed together then Cuo.5Zno.5Fe204 nanoparticles were added to the solution, followed by drop-wise addition of about 2ml Glutaraldehyde and ultrasonication for about 24 hours. After centrifugation, the resulting PEG-CS-Cuo.5Zno.5Fe204 were used for size and zeta potential analysis (dynamic light scattering, DLS) without further processing.Example 2Determination of Cuo.5Zno.sFe204 nanoparticles loading level

[0026] A stock solution of PEG-CS-Cuo.5Zno.5Fe204 nanocomposite was centrifuged, and the resulting sediment was freeze-dried to obtain PEG-CS-Cuo sZno 5Fe204 hybridnanospheres. These hybrid nanospheres were then weighed and subjected to thermal qravimetric analysis. The quantity of unbound Cuo.5Zno.5Fe204 nanoparticles in the supernatant was determined using UV-Visible spectroscopy. The Cuo.5Zno.5Fe204 nanoparticles loading levels were calculated using the following equation:Cu0.5Zn0.5Fe204 nanoparticles loading content weight of Cu0.5Zn0.5Fe204 nanoparticles in the hybrid nanospheres weight of the hybrid nanoshperes x 100%Example 3Evaluating cytotoxicity and anticancer potential

[0027] To explore the potential of the Cuo.5Zno.5Fe204 nanoparticles in anticancer applications, firstly, the size distribution of the nanoparticles was assessed using Polydispersity Index (“PDI”). Subsequently, tests on various cancer and normal human cell lines were conducted, including K562 human leukemia, MDA-MB-231 human breast cancer, MCF7 human breast cancer cell line, and human skin fibroblast. All the cells were cultured and maintained in a humidified incubator at about 37 °C with about 5% CO2. The RPMI-1640 culture medium was used for K562 and MCF7 cells, while MDA-MB-231 and fibroblast were cultured in Dulbecco's Modified Eagle Medium (“DMEM”). Both types of culture media were supplemented with about 10% fetal bovine serum, about 1% penicillin streptomycin, and about 1% L-Glutamine.

[0028] Cells were seeded in a 96-well plate at different seeding densities: 7* 103for MDA- MB- 231 and MCF7, 10* 103for fibroblast, and 15* 103for K562. To facilitate cell attachment, the cells were exposed to a serial dilution of the prepared delivery system(PEG- CS- Cuo.5Zno.5Fe204) after an overnight incubation, ranging from the highest concentration of about 100 pg / ml to the lowest concentration of about 1.6 pg / ml.

[0029] A stock solution of PEG-CS-Cuo.5Zno.5Fe204 nanoparticles at a concentration of 10 mg / ml was made. The nanoparticles were dissolved in a mixture of about 99.5% dimethyl sulfoxide (“DMSO”) and about 0.5% of about 5M nitric acid to enhance their solubility. Subsequently, ultrasonication was performed for about 30 minutes at room temperature. The DMSO concentration in the treatments did not exceed 1%, which is known to be safe and approved for cells, considering DMSO toxicity. After about 72 hours of treatment, theMTT cell viability assay was conducted using the Cell Titer 96® Non-Radioactive Cell Proliferation Assay, and the optical densities (O.Ds) were measured at about 570 nm.Example 4X-ray Diffraction (“XRD”) and Scanning Electron Microscope (“SME”)

[0030] The XRD patterns of all copper-zinc ferrite nanoparticles Cuo.5Zno.5Fe204 samples showed peaks that align well and are in good agreement with standard patterns. Analysis of the Cuo.5Zno.5Fe204 nanoparticles reveals an average crystallite size and lattice constant of the nanoparticles of approximately 85 nm. These values were determined from the most prominent peak (311) using the Scherrer formula. The XRD analysis confirms that the Cuo.5Zno.5Fe204 nanoparticles possess a single-phase spinel structure, displaying no ambiguous similarity and matching the standard pattern of Cuo.5Zno.5Fe204 phase (JCPDS: 00-051-0386) and CuO (JCPDS: 00-041-0254). The experimentally observed d-spacing values and relative intensities correspond well with literature documented values. The XRD spectra show d-values of 2.98, 2.54, and 2.11, which closely conform to the standard XRD pattern. The lattice parameters were within the expected range, confirming the composition of the CuxZn(i-X)Fe2O4 nanoparticles. The diffraction peaks indicate the presence of various phases, including the PEG phase, showing good agreement with the standard pattern JCPDS: 00-050-2158 at 23.48° and 26.1°, the chitosan phase with JCPDS: 00-039-1894 at 20.4°, 24°, and 26.1°, the Cu0.5Zn0.5Fe204 phase with JCPDS: 00-051-0386 at 30.0°, 35.4°, 43.1°, 53.5°, 57.0°, and 62.4°, as well as the ZnO with JCPDS: 01-079-0206 phase and CuO phase with JCPDS: 00-041-0254.

[0031] Comparing the XRD pattern of the Cuo.5Zno.5Fe204 nanoparticles (FIG. la) with the structural patterns of the capping agent (FIG. lb) and (FIG. 1c) attached to the surface of the core of Cuo.5Zno.5Fe204 nanoparticles, which were capped with PEG-CS, respectively, reveals a weakening in the intensity of the core peaks. This is attributed to the capping agent masking the metallic nanoparticles core.

[0032] Based on an average of approximately 200 particles, an SEM image was taken for uncoated Cuo.5Zno.5Fe204 nanoparticles (FIG. 2) and PEG-CS-Cuo sZno sFe2O4 nanoparticles (FIG. 3). The prepared nanoparticles exhibit a spherical shape, with average diameters of 85.1 nm and 340 nm, respectively.Example 5Size analysis, surface charge measurement, and stability studies under variable pH, CaCl2 concentration conditions

[0033] A range of polymeric particle systems with diverse surface charges was evaluated, as an increase in particle charge led to electrostatic interactions between the particle surface and other components, particularly the Cu0.5Zno.5Fe204 nanopatricles, thereby enhancing stability in terms of pH and CaCh that was utilized due to its chelating behavior with the coating polymer linkers. This comparison was performed on the same polymeric coating with different z-potential values, as indicated in stability tests. As shown in Table 1, the size of coated nanoparticles was generated and their stability was studied under different conditions. When the same coating type (“PEG”) was compared for two size ranges for Cuo.5Zno.5Fe204 nanoparticles with and without proscillaridin, it was observed that larger sizes in both cases exhibited lower stability in relation to pH and CaCE concentration, as they aggregated at lower pH values and CaCb concentrations (aggregation occurred at pH 6.4 and at 0.2 M CaCh or higher concentrations). The type of coating polymer used in nanoparticles fabrication significantly affected the Z-potential. The zeta potential value was used to estimate the surface charge and thus the stability of the synthesized nanoparticles. In this context; the zeta potential value on the NPs surface was + 28.5 and 25.3 mV, indicating that the PEG-CS-Cuo sZno sFe2O4 and CS-Cuo.5Zno.5Fe204 have high stability due to higher electrostatic repulsion when compared to uncoated.

[0034] Table 1*Each point represents at least duplicate measurements ± standard deviation. ND: not studied— : aggregation occurs

[0035] While embodiments of the present disclosure have been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various additions, omissions, and modifications can be made without departing from the spirit and scope thereof.

Claims

CLAIMSWhat is claimed is:1- A drug delivery system comprising proscillaridin encapsulated in trimetallic nanoparticles composite and coated with coating polymers.2- The drug delivery system of claim 1, wherein the trimetallic nanoparticles comprise the formula MlxM2yFe2O43- The drug delivery system of claim 2, wherein Ml and M2 are selected from a group comprising Zn+2, Ni+2, Al+3, Mn+2, Ag+, Y+3, Nd+3, Ti+4, Cd+2, Dy+3, Gd+3, Cu+2, Yb+3, Eu+3, Zr+4, In+3, Cr+2, Pr+3, Sm+3, Ho+3, Er+3, Mg+2, La+3, and Ce+34- The drug delivery system of claim 2, wherein x and y have numeric value between 0 and 1.5- The drug delivery system of claim 4, wherein the total sum of x and y equals 1.6- The drug delivery system of claim 1, wherein the coating polymers comprising polyethylene glycol and chitosan.

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