Formulation and Evaluation of α-Pinene Loaded Self-emulsifying Nanoformulation for In-Vivo Anti-Parkinson's Activity

Page: [139 - 159] Pages: 21

  • * (Excluding Mailing and Handling)

Abstract

Aim: The present study was aimed to developed and optimize the self-nano emulsifying drug delivery system of α-pinene (ALP-SNEDDS) and evaluate its in-vivo anti-Parkinson's activity.

Background: Different lipid-based drug delivery technologies have been researched to upgrade drug bioavailability and expand their clinical adequacy upon oral administration. Self-emulsifying drug delivery systems (SEDDS) have pulled in developing the interest specifically for self nano emulsifying drug delivery systems (SNEDDS).

Objective: The present work was attempted to improve the bioavailability of the ALP by defining the role of self-nano emulsifying formulations for its neuroprotective effect.

Methods: Miscibility of the ALP was estimated in various excipient components to select the optimized combination. Self-nano emulsification, thermodynamic stability, the effect of dilution on robustness, optical clarity, viscosity, and conductivity tests were performed. The in-vivo anti-Parkinson's activity of the ALP-SNEDDS formulations were done using Pilocarpine antagonism induced Parkinsonism in rodents. Behavioral tests like tremulous jaw movements, body temperature, salivation, and lacrimation are performed.

Results: Two optimized formulations, composed of Anise oil, Tween 80, and Transcutol-HP of Oil: Smix ratio (4:6 and 3:7) were selected. The Smix ratio for both the formulation was 2:1. The particle size was found to consistent with the increase in dilution. The mean negative zeta potential of the formulations was found to be increased with an increase in dilution. The TEM images of the formulations revealed spherical shape of the droplet. The in-vitro drug release profile was found to be significant as compared to plain ALP suspension.

Conclusion: The results of in-vivo studies indicate that nanosizing and enhanced solubilization of oral ALP-SNEDDS formulations significantly improved the behavioral activities compared to plain ALP suspension.

Keywords: Self-nano emulsifying drug delivery system, alpha pinene, anti Parkinson's, in-vitro, in-vivo, pseudo-ternary phase diagram, blood-brain barrier.

Graphical Abstract

[1]
Naik RR, Shakya AK, Khalaf NA, Abuhamdah S, Oriquat GA, Maraqa A. GC-MS analysis and biological evaluation of essential oil of Zanthoxylum rhesta (roxb.) dc pericarp. Jordan J Pharm Sci 2015; 8(3): 181-93.
[http://dx.doi.org/10.12816/0030449]
[2]
Choudhary KM, Mishra A, Poroikov VV, Goel RK. Ameliorative effect of curcumin on seizure severity, depression like behavior, learning and memory deficit in post-pentylenetetrazole-kindled mice. Eur J Pharmacol 2013; 704(1-3): 33-40.
[http://dx.doi.org/10.1016/j.ejphar.2013.02.012] [PMID: 23461849]
[3]
Sanmukhani J, Satodia V, Trivedi J, et al. Efficacy and safety of curcumin in major depressive disorder: A randomized controlled trial. Phytother Res 2014; 28(4): 579-85.
[http://dx.doi.org/10.1002/ptr.5025] [PMID: 23832433]
[4]
Zhang L, Luo J, Zhang M, Yao W, Ma X, Yu SY. Effects of curcumin on chronic, unpredictable, mild, stress-induced depressive-like behaviour and structural plasticity in the lateral amygdala of rats. Int J Neuropsychopharmacol 2014; 17(5): 793-806.
[http://dx.doi.org/10.1017/S1461145713001661] [PMID: 24405689]
[5]
Kazi M, Shahba AA, Alrashoud S, Alwadei M, Sherif AY, Alanazi FK. Bioactive self-nanoemulsifying drug delivery systems (Bio-SNEDDS) for combined oral delivery of curcumin and piperine. Molecules 2020; 25(7): 1703.
[http://dx.doi.org/10.3390/molecules25071703] [PMID: 32276393]
[6]
Lee GY, Lee C, Park GH, Jang JH. Amelioration of scopolamine-induced learning and memory impairment by α-pinene in C57BL/6 mice. Evid Based Complement Alternat Med 2017; 2017(Oct): 4926815.
[http://dx.doi.org/10.1155/2017/4926815] [PMID: 29234406]
[7]
Kasuya H, Okada N, Kubohara M, Satou T, Masuo Y, Koike K. Expression of BDNF and TH mRNA in the brain following inhaled administration of α-pinene. Phytother Res 2015; 29(1): 43-7.
[http://dx.doi.org/10.1002/ptr.5224] [PMID: 25230317]
[8]
Miyazawa M, Yamafuji C. Inhibition of acetylcholinesterase activity by bicyclic monoterpenoids. J Agric Food Chem 2005; 53(5): 1765-8.
[http://dx.doi.org/10.1021/jf040019b] [PMID: 15740071]
[9]
Chang HJ, Kim HJ, Chun HS. Quantitative Structure-Activity Relationship (QSAR) for neuroprotective activity of terpenoids. Life Sci 2007; 80(9): 835-41.
[http://dx.doi.org/10.1016/j.lfs.2006.11.009] [PMID: 17166521]
[10]
Astani A, Reichling J, Schnitzler P. Comparative study on the antiviral activity of selected monoterpenes derived from essential oils. Phytother Res 2010; 24(5): 673-9.
[http://dx.doi.org/10.1002/ptr.2955] [PMID: 19653195]
[11]
Porres-Martínez M, González-Burgos E, Carretero ME, Gómez-Serranillos MP. In vitro neuroprotective potential of the monoterpenes α-pinene and 1,8-cineole against H2O2-induced oxidative stress in PC12 cells. Z Natforsch C J Biosci 2016; 71(7-8): 191-9.
[http://dx.doi.org/10.1515/znc-2014-4135] [PMID: 27352445]
[12]
Khoshnazar M, Parvardeh S, Bigdeli MR. Alpha-pinene exerts neuroprotective effects via anti-inflammatory and anti-apoptotic mechanisms in a rat model of focal cerebral ischemia-reperfusion. J Stroke Cerebrovasc Dis 2020; 29(8): 104977.
[http://dx.doi.org/10.1016/j.jstrokecerebrovasdis.2020.104977] [PMID: 32689608]
[13]
Khoshnazar M, Bigdeli MR, Parvardeh S, Pouriran R. Attenuating effect of α-pinene on neurobehavioural deficit, oxidative damage and inflammatory response following focal ischaemic stroke in rat. J Pharm Pharmacol 2019; 71(11): 1725-33.
[http://dx.doi.org/10.1111/jphp.13164] [PMID: 31523814]
[14]
Aydin E, Türkez H, Geyikoğlu F. Antioxidative, anticancer and genotoxic properties of α-pinene on N2a neuroblastoma cells. Biologia 2013; 68(5): 1004-9.
[http://dx.doi.org/10.2478/s11756-013-0230-2]
[15]
Türkez H, Aydın E. In vitro assessment of cytogenetic and oxidative effects of α-pinene. Toxicol Ind Health 2016; 32(1): 168-76.
[http://dx.doi.org/10.1177/0748233713498456] [PMID: 24081629]
[16]
Juergens LJ, Tuleta I, Stoeber M, Racké K, Juergens UR. Regulation of monocyte redox balance by 1, 8-cineole (eucalyptol) controls oxidative stress and pro-inflammatory responses in vitro: A new option to increase the antioxidant effects of combined respiratory therapy with budesonide and formoterol? Synergy 2018; 7: 1-9.
[http://dx.doi.org/10.1016/j.synres.2018.05.001]
[17]
Porres-Martínez M, González-Burgos E, Carretero ME, Gómez-Serranillos MP. Major selected monoterpenes α-pinene and 1,8-cineole found in Salvia lavandulifolia (Spanish sage) essential oil as regulators of cellular redox balance. Pharm Biol 2015; 53(6): 921-9.
[http://dx.doi.org/10.3109/13880209.2014.950672] [PMID: 25474583]
[18]
Rufino AT, Ribeiro M, Judas F, et al. Anti-inflammatory and chondroprotective activity of (+)-α-pinene: Structural and enantiomeric selectivity. J Nat Prod 2014; 77(2): 264-9.
[http://dx.doi.org/10.1021/np400828x] [PMID: 24455984]
[19]
Perry NS, Bollen C, Perry EK, Ballard C. Salvia for dementia therapy: Review of pharmacological activity and pilot tolerability clinical trial. Pharmacol Biochem Behav 2003; 75(3): 651-9.
[http://dx.doi.org/10.1016/S0091-3057(03)00108-4] [PMID: 12895683]
[20]
Schmidt L, Göen T. Human metabolism of α-pinene and metabolite kinetics after oral administration. Arch Toxicol 2017; 91(2): 677-87.
[http://dx.doi.org/10.1007/s00204-015-1656-9] [PMID: 26679931]
[21]
Kohlert C, van Rensen I, März R, Schindler G, Graefe EU, Veit M. Bioavailability and pharmacokinetics of natural volatile terpenes in animals and humans. Planta Med 2000; 66(6): 495-505.
[http://dx.doi.org/10.1055/s-2000-8616] [PMID: 10985073]
[22]
Schwarz J, Weisspapir M. Ophthalmic preparation containing menthyl ester of indomethacin United States patent US 8,097,646, 2012.
[23]
Yadav K, Chauhan NS, Saraf S, Singh D, Singh MR. Challenges and need of delivery carriers for bioactives and biological agents: an introduction Advances and avenues in the development of novel carriers for bioactives and biological agents. Academic Press 2020; pp. 1-36.
[http://dx.doi.org/10.1016/B978-0-12-819666-3.00001-8]
[24]
Singh MR, Singh D, Chauhan NS, Kanwar J, Eds. Advances and avenues in the development of novel carriers for bioactives and biological agents Academic Press: Cambredge,. 2020.
[25]
Simona AD, Florina A, Rodica CA, Evelyne O, Maria-Corina S. Nanoscale delivery systems: actual and potential applications in the natural products industry. Curr Pharm Des 2017; 23(17): 2414-21.
[http://dx.doi.org/10.2174/1381612823666170220155540] [PMID: 28228070]
[26]
Rai VK, Narang RK, Pottoo FH, Barkat MA. Pharmacokinetics, interaction, and toxicological profile of nanophytomedicine nanophytomedicine. Singapore: Springer 2020; pp. 133-49.
[27]
Rajagopal S, Ponnusamy M. Channelopathies: Application of natural products using nanotechnology Calcium signaling: From physiology to diseases. Singapore: Springer 2017; pp. 73-86.
[http://dx.doi.org/10.1007/978-981-10-5160-9_6]
[28]
Karpuz M, Gunay MS, Ozer AY. Liposomes and phytosomes for phytoconstituents. In: Advances and avenues in the development of novel carriers for bioactives and biological agents. Amsterdam: Academic Press 2020; pp. 525-53.
[http://dx.doi.org/10.1016/B978-0-12-819666-3.00018-3]
[29]
Zhao Y, Wang C, Chow AH, et al. Self-Nanoemulsifying Drug Delivery System (SNEDDS) for oral delivery of Zedoary essential oil: formulation and bioavailability studies. Int J Pharm 2010; 383(1-2): 170-7.
[http://dx.doi.org/10.1016/j.ijpharm.2009.08.035] [PMID: 19732813]
[30]
Khoo SM, Humberstone AJ, Porter CJ, Edwards GA, Charman WN. Formulation design and bioavailability assessment of lipidic self-emulsifying formulations of halofantrine. Int J Pharm 1998; 167(1-2): 155-64.
[http://dx.doi.org/10.1016/S0378-5173(98)00054-4]
[31]
Shafiq S, Shakeel F, Talegaonkar S, Ahmad FJ, Khar RK, Ali M. Development and bioavailability assessment of ramipril nanoemulsion formulation. Eur J Pharm Biopharm 2007; 66(2): 227-43.
[http://dx.doi.org/10.1016/j.ejpb.2006.10.014] [PMID: 17127045]
[32]
El-Badry M, Haq N, Fetih G, Shakeel F. Solubility and dissolution enhancement of tadalafil using self-nanoemulsifying drug delivery system. J Oleo Sci 2014; 63(6): 567-76.
[http://dx.doi.org/10.5650/jos.ess13236] [PMID: 24770562]
[33]
Kassem AA, Mohsen AM, Ahmed RS, Essam TM. Self-nanoemulsifying drug delivery system (SNEDDS) with enhanced solubilization of nystatin for treatment of oral candidiasis: Design, optimization, in vitro and in vivo evaluation. J Mol Liq 2016; 218: 219-32.
[http://dx.doi.org/10.1016/j.molliq.2016.02.081]
[34]
Zargar-Shoshtari S, Wen J, Alany RG. Formulation and physicochemical characterization of imwitor 308 based self microemulsifying drug delivery systems. Chem Pharm Bull (Tokyo) 2010; 58(10): 1332-8.
[http://dx.doi.org/10.1248/cpb.58.1332] [PMID: 20930400]
[35]
Pouton CW. Lipid formulations for oral administration of drugs: non-emulsifying, self-emulsifying and ‘self-microemulsifying’ drug delivery systems. Eur J Pharm Sci 2000; 11(Suppl. 2): S93-8.
[http://dx.doi.org/10.1016/S0928-0987(00)00167-6] [PMID: 11033431]
[36]
Yang SC, Benita S. Enhanced absorption and drug targeting by positively charged submicron emulsions. Drug Dev Res 2000; 50(3‐4): 476-86.
[http://dx.doi.org/10.1002/1098-2299(200007/08)50:3/4<476:AID-DDR31>3.0.CO;2-6]
[37]
Yang S, Gursoy RN, Lambert G, Benita S. Enhanced oral absorption of paclitaxel in a novel self-microemulsifying drug delivery system with or without concomitant use of P-glycoprotein inhibitors. Pharm Res 2004; 21(2): 261-70.
[http://dx.doi.org/10.1023/B:PHAM.0000016238.44452.f1] [PMID: 15032307]
[38]
Wang D, Chi DF. Morphology and release profile of microcapsules encapsulated alpha-pinene by complex Coacervation. Adv Mat Res 2013; 602: 1285-8.
[39]
Zhao T. Self-Nanoemulsifying Drug Delivery Systems (SNEDDS) for the oral delivery of lipophilic drugs. [PhD thesis], University of Trento, 2015.
[40]
[41]
Vogel HG, Ed. Drug discovery and evaluation: Pharmacological assays. Springer Science & Business Media 2002.
[http://dx.doi.org/10.1007/3-540-29837-1]
[42]
Podurgiel S, Collins-Praino LE, Yohn S, et al. Tremorolytic effects of safinamide in animal models of drug-induced parkinsonian tremor. Pharmacol Biochem Behav 2013; 105: 105-11.
[http://dx.doi.org/10.1016/j.pbb.2013.01.015] [PMID: 23360954]
[43]
Salamone JD, Mayorga AJ, Trevitt JT, Cousins MS, Conlan A, Nawab A. Tremulous jaw movements in rats: a model of parkinsonian tremor. Prog Neurobiol 1998; 56(6): 591-611.
[http://dx.doi.org/10.1016/S0301-0082(98)00053-7] [PMID: 9871939]
[44]
Singh AK, Chaurasiya A, Awasthi A, et al. Oral bioavailability enhancement of exemestane from Self-Microemulsifying Drug Delivery System (SMEDDS). AAPS PharmSciTech 2009; 10(3): 906-16.
[http://dx.doi.org/10.1208/s12249-009-9281-7] [PMID: 19609837]
[45]
Balakumar K, Raghavan CV, Selvan NT, Prasad RH, Abdu S. Self Nanoemulsifying Drug Delivery System (SNEDDS) of rosuvastatin calcium: design, formulation, bioavailability and pharmacokinetic evaluation. Colloids Surf B Biointerfaces 2013; 112: 337-43.
[http://dx.doi.org/10.1016/j.colsurfb.2013.08.025] [PMID: 24012665]
[46]
Parmar N, Singla N, Amin S, Kohli K. Study of cosurfactant effect on nanoemulsifying area and development of lercanidipine loaded (SNEDDS) Self Nanoemulsifying Drug Delivery System. Colloids Surf B Biointerfaces 2011; 86(2): 327-38.
[http://dx.doi.org/10.1016/j.colsurfb.2011.04.016] [PMID: 21550214]
[47]
Fahmy UA, Ahmed OA, Hosny KM. Development and evaluation of avanafil self-nanoemulsifying drug delivery system with rapid onset of action and enhanced bioavailability. AAPS PharmSciTech 2015; 16(1): 53-8.
[http://dx.doi.org/10.1208/s12249-014-0199-3] [PMID: 25168449]
[48]
Hosny KM, Banjar ZM. The formulation of a nasal nanoemulsion zaleplon in situ gel for the treatment of insomnia. Expert Opin Drug Deliv 2013; 10(8): 1033-41.
[http://dx.doi.org/10.1517/17425247.2013.812069] [PMID: 23795561]
[49]
Constantinides PP. Lipid microemulsions for improving drug dissolution and oral absorption: Physical and biopharmaceutical aspects. Pharm Res 1995; 12(11): 1561-72.
[http://dx.doi.org/10.1023/A:1016268311867] [PMID: 8592652]
[50]
Hathout RM, Woodman TJ, Mansour S, Mortada ND, Geneidi AS, Guy RH. Microemulsion formulations for the transdermal delivery of testosterone. Eur J Pharm Sci 2010; 40(3): 188-96.
[http://dx.doi.org/10.1016/j.ejps.2010.03.008] [PMID: 20304048]
[51]
Baker RC, Florence AT, Ottewill RH, Tadros TF. Investigations into the formation and characterization of microemulsions. II. Light scattering conductivity and viscosity studies of microemulsions. J Colloid Interface Sci 1984; 100(2): 332-49.
[http://dx.doi.org/10.1016/0021-9797(84)90439-9]
[52]
Singh AK, Chaurasiya A, Singh M, Upadhyay SC, Mukherjee R, Khar RK. Exemestane loaded Self-Microemulsifying Drug Delivery System (SMEDDS): development and optimization. AAPS PharmSciTech 2008; 9(2): 628-34.
[http://dx.doi.org/10.1208/s12249-008-9080-6] [PMID: 18473177]
[53]
Shah NH, Carvajal MT, Patel CI, Infeld MH, Malick AW. Self-Emulsifying Drug Delivery Systems (SEDDS) with polyglycolyzed glycerides for improving in vitro dissolution and oral absorption of lipophilic drugs. Int J Pharm 1994; 106(1): 15-23.
[http://dx.doi.org/10.1016/0378-5173(94)90271-2]
[54]
Agrawal AG, Kumar A, Gide PS. Formulation of solid self-nanoemulsifying drug delivery systems using N-methyl pyrrolidone as cosolvent. Drug Dev Ind Pharm 2015; 41(4): 594-604.
[http://dx.doi.org/10.3109/03639045.2014.886695] [PMID: 24517575]
[55]
Badran MM, Taha EI, Tayel MM, Al-Suwayeh SA. Ultra-fine self nanoemulsifying drug delivery system for transdermal delivery of meloxicam: Dependency on the type of surfactants. J Mol Liq 2014; 190: 16-22.
[http://dx.doi.org/10.1016/j.molliq.2013.10.015]
[56]
Choi KO, Aditya NP, Ko S. Effect of aqueous pH and electrolyte concentration on structure, stability and flow behavior of non-ionic surfactant based solid lipid nanoparticles. Food Chem 2014; 147: 239-44.
[http://dx.doi.org/10.1016/j.foodchem.2013.09.095] [PMID: 24206712]
[57]
Singh SK, Verma PR, Razdan B. Glibenclamide-loaded self-nanoemulsifying drug delivery system: development and characterization. Drug Dev Ind Pharm 2010; 36(8): 933-45.
[http://dx.doi.org/10.3109/03639040903585143] [PMID: 20184416]
[58]
Gupta S, Kesarla R, Omri A. Formulation strategies to improve the bioavailability of poorly absorbed drugs with special emphasis on self-emulsifying systems. Int Sch Res Notices 2013; Article ID 848043..
[http://dx.doi.org/10.1155/2013/848043]
[59]
Lawrence MJ, Rees GD. Microemulsion-based media as novel drug delivery systems. Adv Drug Deliv Rev 2000; 45(1): 89-121.
[http://dx.doi.org/10.1016/S0169-409X(00)00103-4] [PMID: 11104900]
[60]
Dixit AR, Rajput SJ, Patel SG. Preparation and bioavailability assessment of SMEDDS containing valsartan. AAPS PharmSciTech 2010; 11(1): 314-21.
[http://dx.doi.org/10.1208/s12249-010-9385-0] [PMID: 20182825]
[61]
Shakeel F, Haq N, Alanazi FK, Alsarra IA. Polymeric solid self-nanoemulsifying drug delivery system of glibenclamide using coffee husk as a low cost biosorbent. Powder Technol 2014; 256: 352-60.
[http://dx.doi.org/10.1016/j.powtec.2014.02.028]
[62]
Xi J, Chang Q, Chan CK, et al. Formulation development and bioavailability evaluation of a self-nanoemulsified drug delivery system of oleanolic acid. AAPS PharmSciTech 2009; 10(1): 172-82.
[http://dx.doi.org/10.1208/s12249-009-9190-9] [PMID: 19224372]
[63]
Elnaggar YS, El-Massik MA, Abdallah OY. Self-nanoemulsifying drug delivery systems of tamoxifen citrate: Design and optimization. Int J Pharm 2009; 380(1-2): 133-41.
[http://dx.doi.org/10.1016/j.ijpharm.2009.07.015] [PMID: 19635537]
[64]
Zhang P, Liu Y, Feng N, Xu J. Preparation and evaluation of self-microemulsifying drug delivery system of oridonin. Int J Pharm 2008; 355(1-2): 269-76.
[http://dx.doi.org/10.1016/j.ijpharm.2007.12.026] [PMID: 18242895]
[65]
Martín MJ, Calpena AC, Fernández F, Mallandrich M, Gálvez P, Clares B. Development of alginate microspheres as nystatin carriers for oral mucosa drug delivery. Carbohydr Polym 2015; 117: 140-9.
[http://dx.doi.org/10.1016/j.carbpol.2014.09.032] [PMID: 25498619]
[66]
Marín-Quintero D, Fernández-Campos F, Calpena-Campmany AC, Montes-López MJ, Clares-Naveros B, Del Pozo-Carrascosa A. Formulation design and optimization for the improvement of nystatin-loaded lipid intravenous emulsion. J Pharm Sci 2013; 102(11): 4015-23.
[http://dx.doi.org/10.1002/jps.23711] [PMID: 23970386]
[67]
Fernández-Campos F, Clares Naveros B, López Serrano O, Alonso Merino C, Calpena Campmany AC. Evaluation of novel nystatin nanoemulsion for skin candidosis infections. Mycoses 2013; 56(1): 70-81.
[http://dx.doi.org/10.1111/j.1439-0507.2012.02202.x] [PMID: 22574899]
[68]
Fernández Campos F, Calpena Campmany AC, Rodríguez Delgado G, López Serrano O, Clares Naveros B. Development and characterization of a novel nystatin-loaded nanoemulsion for the buccal treatment of candidosis: Ultrastructural effects and release studies. J Pharm Sci 2012; 101(10): 3739-52.
[http://dx.doi.org/10.1002/jps.23249] [PMID: 22777575]
[69]
Martín-Villena MJ, Fernández-Campos F, Calpena-Campmany AC, Bozal-de Febrer N, Ruiz-Martínez MA, Clares-Naveros B. Novel microparticulate systems for the vaginal delivery of nystatin: Development and characterization. Carbohydr Polym 2013; 94(1): 1-11.
[http://dx.doi.org/10.1016/j.carbpol.2013.01.005] [PMID: 23544502]
[70]
Heshmati N, Cheng X, Eisenbrand G, Fricker G. Enhancement of oral bioavailability of E804 by Self-Nanoemulsifying Drug Delivery System (SNEDDS) in rats. J Pharm Sci 2013; 102(10): 3792-9.
[http://dx.doi.org/10.1002/jps.23696] [PMID: 23934779]
[71]
Thomas N, Müllertz A, Graf A, Rades T. Influence of lipid composition and drug load on the in vitro performance of self-nanoemulsifying drug delivery systems. J Pharm Sci 2012; 101(5): 1721-31.
[http://dx.doi.org/10.1002/jps.23054] [PMID: 22294458]
[72]
Beg S, Swain S, Singh HP, Patra ChN, Rao ME. Development, optimization, and characterization of solid self-nanoemulsifying drug delivery systems of valsartan using porous carriers. AAPS PharmSciTech 2012; 13(4): 1416-27.
[http://dx.doi.org/10.1208/s12249-012-9865-5] [PMID: 23070560]
[73]
Kanuganti S, Jukanti R, Veerareddy PR, Bandari S. Paliperidone-loaded Self-Emulsifying Drug Delivery Systems (SEDDS) for improved oral delivery. J Dispers Sci Technol 2012; 33(4): 506-15.
[http://dx.doi.org/10.1080/01932691.2011.574920]
[74]
Pomara N, Block R, Demetriou S, Fucek F, Stanley M, Gershon S. Attenuation of pilocarpine-induced hypothermia in response to chronic administration of choline. Psychopharmacology (Berl) 1983; 80(2): 129-30.
[http://dx.doi.org/10.1007/BF00427955] [PMID: 6410440]
[75]
Rupniak NM, Jenner P, Marsden CD. Cholinergic manipulation of perioral behaviour induced by chronic neuroleptic administration to rats. Psychopharmacology (Berl) 1983; 79(2-3): 226-30.
[http://dx.doi.org/10.1007/BF00427817] [PMID: 6133305]
[76]
Salamone JD, Lalies MD, Channell SL, Iversen SD. Behavioural and pharmacological characterization of the mouth movements induced by muscarinic agonists in the rat. Psychopharmacology (Berl) 1986; 88(4): 467-71.
[http://dx.doi.org/10.1007/BF00178508] [PMID: 3085134]
[77]
Stewart BR, Jenner P, Marsden CD. The pharmacological characterisation of pilocarpine-induced purposeless chewing behaviour in the rat. Psychopharmacology (Berl) 1988; 96(1): 55-62.
[http://dx.doi.org/10.1007/BF02431533] [PMID: 2906443]