RESEARCH ARTICLE – Pharmaceutical Nanotechnology
Intranasal Piperine-Loaded Chitosan Nanoparticles as Brain-Targeted Therapy in Alzheimer's Disease: Optimization, Biological Efficacy, and Potential Toxicity

https://doi.org/10.1002/jps.24557Get rights and content

ABSTRACT

Piperine (PIP) is a phytopharmaceutical with reported neuroprotective potential in Alzheimer's disease (AD). Oral PIP delivery suffers from its hydrophobicity and pre-systemic metabolism. In this article, mono-disperse intranasal chitosan nanoparticles (CS-NPs) were elaborated for brain targeting of PIP. Formula optimization was based on particle size (PS), zeta potential (ZP), polydispersity index (PDI), % entrapment efficiency (% EE), release studies, and transmission electron microscopy. AD was induced in 48 male Wistar rats on which full behavioral and biochemical testing was conducted. Brain toxicity was assessed based on Caspase-3 assay for apoptosis and tumor necrosis factor for inflammation. Spherical NPs with optimum % EE (81.70), PS (248.50 nm), PDI (0.24), and ZP (+ 56.30 mV) were elaborated. PIP-NPs could significantly improve cognitive functions as efficient as standard drug (donpezil injection) with additional advantages of dual mechanism (Ach esterase inhibition and antioxidant effect). CS-NPs could significantly alleviate PIP nasal irritation and showed no brain toxicity. This work was the first to report additional mechanism of PIP in AD via anti-apoptosis and anti-inflammatory effects. To conclude, mucoadhesive CS-NPs were successfully tailored for effective, safe, and non-invasive PIP delivery with 20-folds decrease in oral dose, opening a gate for a future with lower AD morbidity. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:3544–3556, 2015

Section snippets

INTRODUCTION

Alzheimer’s disease (AD) is the most common cause of dementia worldwide; it affects more than 35 million people, and this number is expected to triple in the next 40 years because of the increase in life expectancy.1 AD is a neurodegenerative disorder that is characterized by not only progressive impairment of memory and cognitive functions but also disturbances in behavior.2., 3.

A smaller proportion (5%–10%) of all Alzheimer cases was reported to be caused by genetic abnormalities of

Materials

Chitosan with medium molecular weight (Mw = 100,000–300,000 Da) and degree of deacetylation > 90% and PIP (Mw = 285.34 Da, purity 98%) were purchased from Alpha Aeser (Ward Hill, MA 01835, United States, Massachusetts), sodium TPP was purchased from Sigma–Aldrich (St. Louis, Missouri). Poloxamer 188, Tween 80, glacial acetic acid, methanol AR, and all other reagents were of analytical grade. Colchicine, donepezil hydrochloride, thiobarbituric acid (TBA), trichloracetic acid (TCA), nitroblue

Preparation and Characterization of CS-NPs

Elaboration of various nanocarriers for solving drug delivery obstacles have been recently adopted by our work group31., 36., 37., 53., 54., 55., 56., 57., 58., 59. Among others, cross-linked CS-NPs have been rationally selected in this article and prepared by ionic gelation method.33 The latter offers simple and mild preparation conditions in aqueous environment and relies on the pH-dependent electrostatic interaction between positively charged amino groups in CS and the negatively charged

CONCLUSIONS

In the current study, hydrophobic drug PIP could be successively loaded in hydrophilic CS NPs with high entrapment and ZP sufficient for stabilization. Impact of nine optimization factors (CS solution concentration, CS:TPP ratio, stirring rate, stabilizer incorporation, CS temperature, homogenization, loading dose, and loading method) on size tuning of NPs was investigated. Adjusted interplay between such factors could elaborate the desired PS fitting the cut off for brain delivery. PIP

REFERENCES (86)

  • Y. Aktas et al.

    Preparation and in vitro evaluation of chitosan nanoparticles containing a caspase inhibitor

    Int J Pharm

    (2005)
  • M. Fazil et al.

    Development and evaluation of rivastigmine loaded chitosan nanoparticles for brain targeting

    Eur J Pharm Sci

    (2012)
  • R. D’Hooge et al.

    Applications of the Morris water maze in the study of learning and memory

    Brain Res Brain Res Rev

    (2001)
  • M.H. Veerendra Kumar et al.

    Intracerebroventricular administration of colchicine produces cognitive impairment associated with oxidative stress in rats

    Pharmacol Biochem Behav

    (2002)
  • Y. Kono

    Generation of superoxide radical during autoxidation of hydroxylamine and an assay for superoxide dismutase

    Arch Biochem Biophys

    (1978)
  • G.L. Ellman et al.

    A new and rapid colorimetric determination of acetylcholinesterase activity

    Biochem Pharmacol

    (1961)
  • O.H. Lowry et al.

    Protein measurement with the Folin phenol reagent

    J Biol Chem

    (1951)
  • Y.S. Elnaggar et al.

    Lecithin-based nanostructured gels for skin delivery: An update on state of art and recent applications

    J Control Release

    (2014)
  • M.S. Freag et al.

    Development of novel polymer-stabilized diosmin nanosuspensions: In vitro appraisal and ex vivo permeation

    Int J Pharm

    (2013)
  • M.A. Elsheikh et al.

    Rationale employment of cell culture versus conventional techniques in pharmaceutical appraisal of nanocarriers

    J Control Release

    (2014)
  • W.M. El-Refaie et al.

    Novel curcumin-loaded gel-core hyaluosomes with promising burn-wound healing potential: Development, in-vitro appraisal and in-vivo studies

    Int J Pharm

    (2015)
  • Y. Xu et al.

    Effect of molecular structure of chitosan on protein delivery properties of chitosan nanoparticles

    Int J Pharm

    (2003)
  • X. Zhang et al.

    Preparation and characterization of insulin-loaded bioadhesive PLGA nanoparticles for oral administration

    Eur J Pharm Sci

    (2012)
  • G. Riedel et al.

    Glutamate receptor function in learning and memory

    Behav Brain Res

    (2003)
  • V.F. Prado et al.

    Mice deficient for the vesicular acetylcholine transporter are myasthenic and have deficits in object and social recognition

    Neuron

    (2006)
  • J. Dai et al.

    Impaired axonal transport of cortical neurons in Alzheimer’s disease is associated with neuropathological changes

    Brain Res

    (2002)
  • M.R. Roberson et al.

    Cholinergic activity and amyloid precursor protein metabolism

    Brain Res Brain Res Rev

    (1997)
  • A. Kumar et al.

    Differential effects of cyclooxygenase inhibitors on intracerebroventricular colchicine-induced dysfunction and oxidative stress in rats

    Euro J pharmacol

    (2006)
  • S. Md et al.

    Bromocriptine loaded chitosan nanoparticles intended for direct nose to brain delivery: Pharmacodynamic, pharmacokinetic and scintigraphy study in mice model

    Eur J Pharm Sci

    (2013)
  • X. Wang et al.

    Preparation of estradiol chitosan nanoparticles for improving nasal absorption and brain targeting

    Euro J Pharm Biopharm

    (2008)
  • A.J. Friedman et al.

    Antimicrobial and anti-inflammatory activity of chitosan-alginate nanoparticles: A targeted therapy for cutaneous pathogens

    J Invest Dermatol

    (2013)
  • P. Shrivastava et al.

    Anti-apoptotic and anti-inflammatory effect of Piperine on 6-OHDA induced Parkinson’s rat model

    J Nutr Biochem

    (2013)
  • M. Prince et al.

    The global prevalence of dementia: A systematic review and metaanalysis

    Alzheimers Dement

    (2013)
  • A.K. Desai et al.

    Behavioral disturbance in dementia

    Curr Psychiatry Rep

    (2012)
  • L.E. Hebert et al.

    Alzheimer disease in the US population: Prevalence estimates using the 2000 census

    Arch Neurol

    (2003)
  • S.S. Matsuyama et al.

    Hypothesis: Microtubules, a key to Alzheimer disease

    Proc Natl Acad Sci USA

    (1989)
  • C.F. Flaherty et al.

    Effects of intrahippocampal administration of colchicine on incentive contrast and on radial maze performance

    Behav Neurosci

    (1989)
  • V. Calabrese et al.

    Nutritional antioxidants and the heme oxygenase pathway of stress tolerance: Novel targets for neuroprotection in Alzheimer’s disease

    Ital J Biochem

    (2003)
  • K. Kontush et al.

    Vitamin E in neurodegenerative disorders: Alzheimer’s disease

    Ann N Y Acad Sci

    (2004)
  • N. Qizilbash et al.

    WITHDRAWN: Tacrine for Alzheimer’s disease

    Cochrane Database Syst Rev

    (2007)
  • R. D’Hooge et al.

    Anticonvulsant activity of piperine on seizures induced by excitatory amino acid receptor agonists

    Arzneimittelforschung

    (1996)
  • L.H. Lai et al.

    Piperine suppresses tumor growth and metastasis in vitro and in vivo in a 4 T1 murine breast cancer model

    Acta Pharmacol Sin

    (2012)
  • S.A. Lee et al.

    Piperine from the fruits of Piper longum with inhibitory effect on monoamine oxidase and antidepressant-like activity

    Chem Pharm Bull (Tokyo)

    (2005)
  • Cited by (244)

    View all citing articles on Scopus
    View full text