Lipid drug conjugate nanoparticle as a potential nanocarrier for the oral delivery of pemetrexed diacid: Formulation design, characterization, ex vivo, and in vivo assessment
Introduction
Pemetrexed diacid (N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo-[2,3-d]-pyrimidin-5-yl)-ethyl]-benzoyl]-l-glutamic acid belongs to a class of chemotherapeutic drugs known as folate antimetabolites [1], [2]. Pemetrexed diacid (PTX) developed by Eli Lilly and Company, was approved by the United States Food and Drug administration in 2004 for the treatment of malignant pleural mesothelioma in combination with cisplatin, a platinum-containing chemotherapeutic drug and a second line agent for the treatment of advanced or metastatic non-small cell lung cancer. Currently, the drug is used as a single agent or in combination with other chemotherapeutic agents for the treatment of other types of cancer such as breast cancer, bladder cancer, colorectal carcinoma and cervical cancer [3], [4]. It is available in the form of intravenous injection, marketed by Eli Lilly as Pemetrexed disodium Injection (Alimta®)100 mg and 500 mg lyophilized Powder. Dosing Regimen followed is 500 mg/m2 i.v. on day 1 of each 21-day cycle [5]. Oral bioavailability has not been evaluated since pemetrexed is intended for use by short-term i.v. infusion. However, oral absorption was evaluated in mice using 20 mg/kg i.v. and oral doses of 14C radiolabeled pemetrexed. Results from mice indicate that the oral absorption is low with only 13% of an oral dose absorbed in mice [6]. Although it is sparingly soluble (0.0905 mg/ml) in water, the permeation of the drug is low, which may be the cause of low oral bioavailability. Till date no oral formulation of the drug has been available [7]. So, we aimed to formulate an oral dosage form which can solve the problem of solubility as well as permeability in intestine and increase the overall oral bioavailability of the drug which can lead to increase patient compliance for the therapy.
Lipid-based nanoparticles represent an alternative drug carrier system to traditional colloidal carriers, such as liposome, nanoemulsions, polymeric nanoparticles and polymeric microparticles, and also they possess advantages of controlled drug release, drug targeting, increase in intestinal permeability, and increase in bioavailability. Moreover, they avoid the need for use of organic solvents [8], [9], [10]. Hence, the current focus of the research based on the search of bio-compatible lipids as a carrier for low bioavailable drugs to minimize problem of the mammalian tissue toxicity due to use of organic solvents, limited physical stability and leakage of drug during storage [11], [12]. Stearic acid has been already used for the delivery of many drugs as lipid-based nanoformulation as well as Lipid drug conjugate (LDC) nanoparticle [13]. The drug molecule shows the presence of carboxylic and amino groups, which is an ideal condition for linking with lipids such as stearic acid. It has been widely reported that drugs incorporated into lipid and stabilized by the use of various surfactants show high permeability of the intestine because lipids and these surfactants act as a good permeation enhancer of drugs from the gastrointestinal tract by solubilization of the drug in the intestinal milieu and reduce the first-pass metabolism of the drug by transport of the drug through a lymphatic route to the systemic circulation [14], [15]. The surfactants (Labrasol, capryol 90 and Tween 80) which are used for the stabilization of the LDC nanoparticle have their own role in the increase in the permeability of the drug through the intestine by inhibiting the P-glycoprotein efflux pump that is present in the intestinal brush border region [16], [17]. These lipid nanoparticles are becoming one of the most acceptable drug delivery systems for both lipophilic as well as hydrophilic drugs because of ease of production, easy scalability and, most advantageously, the use of lipid excipients with generally regarded as safe (GRAS) status, avoiding the use of organic solvents [18].
The literature search revealed no reports on the LDC nanoparticle and other oral dosage forms for PTX. Therefore, the specific objective of this study was to develop a LDC nanoparticle for the oral delivery of PTX by using stearic acid as a lipid using cold homogenization technique. The role of key independent variables influencing on the dependent variables (particle size, polydispersity index and entrapment efficiency) was determined by constructing a Box–Behnken design. Also the characterization of the optimized LDC nanoparticle was made for various parameters and an ex vivo gut permeation study was performed to determine the apparent permeability coefficient of the formulation in comparison with a plain suspension of the PTX. Finally the efficacy of the developed formulation against cancer cells was evaluated.
Section snippets
Material
Pemetrexed diacid (PTX) was obtained as a gift sample from Dabur research foundation, India. Labrasol and capryol 90 were obtained from gattefosse (Mumbai, india) as gift samples. Stearic acid was purchased from qualikem fine chemicals, India. Tween 80 was purchased from S.D fine chemicals, India. Methanol HPLC grade, ortho-phosphoric acid, triethylamine, HPLC grade water and ethanol (SRL, Mumbai, India) were used. All the other reagents used in the study were of analytical grade.
Excipients screening
Stearic acid
Screening of excipients
Selection of stearic acid was made for the production of LDC nanoparticles due to the presence of carboxyl group which helps in conjugating with amino and hydroxyl group of the drug PTX. The surfactant combination selected was tween 80, capryol 90 and labrasol in the ratio 2:1:1. This was selected on the basis of stability of the coarse dispersions.
Optimization of LDC nanoparticles
Box–Behnken statistical screening design was used to statistically optimize the formulation parameters and evaluate main effects, interaction
Conclusion
LDC nanoparticles of PTX were successfully prepared by the cold homogenization technique by using stearic acid as the lipid which successfully conjugated with the PTX. The formulation was optimized by RSM using Design-Expert software, and the optimized batch was characterized for various parameters. The optimized formulation was found to be efficient as suggested by in vitro, ex vivo and in vivo assessments. A much deeper penetration to the intestine was observed with LDC nanoparticles by using
Conflict of interest
The authors report no conflicts of interest.
Acknowledgments
The authors are thankful to the Central Instrumental Facility (CIF) Laboratory, Jamia Hamdard, New Delhi, India to carry out the present research work. The authors would like to convey their heartiest gratitude to Yub Raj Neupane of the Faculty of Pharmacy, Jamia Hamdard, for providing timely support and suggestions related to formulation development.
Reference: (33)
- et al.
Pemetrexed: a multitargeted antifolate
Clin. Ther.
(2005) - et al.
Oral bioavailability of cyclosporine: solid lipid nanoparticles (SLNR) versus drug nanocrystals
Int. J. Pharm.
(2006) - et al.
Lipid nanoparticles for brain targeting I. formulation optimization
Int. J. Pharm.
(2011) - et al.
Lipid-drug conjugate nanoparticles of the hydrophilic drug diminazene-cytotoxicity testing and mouse serum adsorption
J. Control. Release
(2004) - et al.
Statistical optimization and characterization of pH-independent extended-release drug delivery of cefpodoxime proxetil using Box–Behnken design
Chem. Eng. Res. Des.
(2014) - et al.
Optimization and formulation design of gels of Diclofenac and Curcumin for transdermal drug delivery by Box-Behnken statistical design
J. Pharm. Sci.
(2011) - et al.
Formulation of extended release cefpodoxime proxetil chitosan-alginate beads using quality by design approach
Int. J. Biol. Macromol.
(2014) - et al.
Development of topotecan loaded lipid nanoparticles for chemical stabilization and prolonged release
Eur. J. Pharm. Biopharm.
(2011) - et al.
Particle size-dependent organ distribution of gold nanoparticles after intravenous administration
Biomaterials
(2008) - et al.
Lipid based nanocarrier system for the potential oral delivery of decitabine: formulation design, characterization, ex vivo, and in vivo assessment
Int. J. Pharm.
(2014)
Formulation of extended release cefpodoxime proxetil chitosan-alginate beads using quality by design approach
Int. J. Biol. Macromol.
Solvent injection as a new approach for manufacturing lipid nanoparticles—evaluation of the method and process parameters
Eur. J. Pharm. Biopharm.
Oral bioavailability of cyclosporine: solid lipid nanoparticles (SLN) versus drug nanocrystals
Int. J. Pharm.
Antineoplastics and immunosuppressants
Martindale˗The Complete Drug Reference
The Merck Index: An Encyclopedia of Chemicals, Drugs, and Biologicals
Drug Facts and Comparisons
Cited by (35)
Apolipoprotein E3 functionalized lipid-drug conjugated nanoparticles of Levetiracetam for enhanced delivery to the brain: In-vitro cell line studies and in-vivo study
2024, International Journal of Biological MacromoleculesLymphatic transport system to circumvent hepatic metabolism for oral delivery of lipid-based nanocarriers
2021, Journal of Drug Delivery Science and TechnologyCitation Excerpt :MTT assay was carried out on human lung adenocarcinoma cell lines (A 549) and reported that LDC NPs loaded PTX showed significant cytotoxicity at 25, 50, and 100 μg/mL, respectively, compared to plain PTX solution because of the biphasic release performance of PTX from optimized LDC NPs. A pharmacokinetic study was studied using a non-compartmental method and reported that LDC NPs loaded PTX at 20 mg/kg showed significant enhancement of Cmax, Tmax, and AUC0-24 compared to plain PTX solution suggest controlled release of PTX from LDC NPs responsible for the enhancement of oral bioavailability of PTX [135]. Ashwini Kumar et al. have developed 5-fluorouracil-palmitic acid conjugate (5-FUDIPAL) incorporated in polyester nanoparticles using the dual technique, i.e., double emulsion-solvent evaporation method, and evaluated its efficacy for controlled release and cytotoxicity in HCT -116 cell line (colon cancer).
Big family of nano- and microscale drug delivery systems ranging from inorganic materials to polymeric and stimuli-responsive carriers as well as drug-conjugates
2021, Journal of Drug Delivery Science and TechnologyCitation Excerpt :Thus, lipid-nicotine conjugate nanoarchitectures of large loading efficacy produced using hydrogenated sunflower oil (lipid), Kolliwax® S and stearic acid (counter ion) could be used in prevention of lung cancer. Lipid-drug conjugates were prepared as efficient nanomaterials to orally deliver pemetrexed diacid and evaluate their in vivo, ex vivo and in vitro characteristics [292]. For this purpose, pemetrexed diacid salt was formed using stearic acid that was followed via cold homogenization to achieve LDCs nanoformulations.
Oral lipid nanomedicines: Current status and future perspectives in cancer treatment
2021, Advanced Drug Delivery ReviewsRemoval of Pemetrexed from aqueous phase using activated carbons in static mode
2021, Chemical Engineering JournalCitation Excerpt :Kleywegt et al. reported that the use of granulated AC could increase the removal efficiency of carbamazepine from 71 to 93% for water treatment in Ontario, Canada [12]. Pemetrexed (PEME) is a novel generation of anti–folate pharmaceutical showing encouraging activity in the treatment of a variety of tumors, e.g., malignant mesothelioma, non–small cell lung cancer, breast cancer, bladder cancer, colorectal carcinoma and cervical cancer [13]. Consequently, PEME consumption increases year by year, especially in European countries and it is worth noting that PEME environmental concentrations in France for years 2004 and 2008 were reported to be 0.02 and 0.85 ng.L−1, respectively [14].
Nano lipid based carriers for lymphatic voyage of anti-cancer drugs: An insight into the in-vitro, ex-vivo, in-situ and in-vivo study models
2020, Journal of Drug Delivery Science and TechnologyCitation Excerpt :Further, the Papp value for the LDC nanoparticulate system of the drug was found to be 3.056 × 10−4 cm min−1, whereas it was only 0.5082 × 10−4 cm min−1 for the plain drug solution. These findings suggested that the LDC can be effectively used for enhanced intestinal permeability of anti-cancer drugs [124]. More recently Azandaryani et al., 2019 used a non-everted model to assess the permeability coefficients (Papp) of hydrophobic drug letrozole (an aromatase inhibitor) from hybrid lipid-based nanoparticles (chitosan-lipid).