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Plant Science Today (PST)

Research Articles

Early Access

Formulation and characterization of 6,6′-dihydroxythiobinupharidine and phloroglucinol nanoemulsion for intranasal approach to neurodegenerative disorders

DOI
https://doi.org/10.14719/pst.9150
Submitted
28 April 2025
Published
10-11-2025
Versions

Abstract

Neurodegenerative diseases involve complex pathological mechanisms, including oxidative stress and neuroinflammation, which are not adequately addressed by single-target therapies. Therefore, a dual-targeted strategy is to be employed by combining phloroglucinol, a potent antioxidant, with 6,6′-dihydroxythiobinupharidine, a selective protein kinase C inhibitor. Thus, the present study aimed to formulate and evaluate phloroglucinol and 6,6′-dihydroxythiobinupharidine nanoemulsion (NE) for intranasal administration and a dual-targeted approach. NE was formulated using coconut oil, Span80, Tween80 and PEG400 in the ratio of 4.5:4.5:5.5:5.5. After formulation, it was evaluated for size, shape, drug release and stability. NE was homogenously opaque white and clear i.e., without any visible drug particles. It was an oil-in-water type of NE with a circular shape and size of <150 nm. It possessed a polydispersity index of 0.238, a zeta potential of -26.2 mV and was stable for more than 90 days. It was readily dispersible upon addition of water and showed no separation of oil and water. There was no sign of precipitation or cracking and exhibited a percentage transmittance of 93.4 ± 0.67. Nanoemulsified phloroglucinol and 6,6′-dihydroxythiobinupharidine had a cumulative release rate of 88.79 ± 1.92 and 90.39 ± 1.26, respectively, after 2 hr and 1 hr. Over 90 days, an increase in the particle size and a faint odor were observed. Therefore, these results indicate that the NE of phloroglucinol and 6,6′-dihydroxythiobinupharidine enables efficient and targeted delivery of drugs directly to the brain and improves treatment effectiveness for neurodegenerative disorders.

References

  1. 1. Lofts A, Abu-Hijleh F, Rigg N, Mishra RK, Hoare T. Using the intranasal route to administer drugs to treat neurological and psychiatric illnesses: rationale, successes, and future needs. CNS Drugs. 2022;36:739-70. https://doi.org/10.1007/s40263-022-00930-4
  2. 2. Wen P, Ren C. Research progress on intranasal treatment for Parkinson's disease. Neuroprotection. 2024;2(2):79-99. https://doi.org/10.1002/nep3.42
  3. 3. Koo J, Lim C, Oh KT. Recent advances in intranasal administration for brain-targeting delivery: a comprehensive review of lipid-based nanoparticles and stimuli-responsive gel formulations. Int J Nanomedicine. 2024;19:1767-807. https://doi.org/10.2147/IJN.S439181
  4. 4. Keller LA, Merkel O, Popp A. Intranasal drug delivery: opportunities and toxicologic challenges during drug development. Drug Deliv Transl Res. 2022;12:735-57. https://doi.org/10.1007/s13346-020-00891-5
  5. 5. Djupesland PG, Messina JC, Mahmoud RA. The nasal approach to delivering treatment for brain diseases: an anatomic, physiologic, and delivery technology overview. Ther Deliv. 2014;5(6):709-33. https://doi.org/10.4155/tde.14.41
  6. 6. Park C, Cha HJ, Hwangbo H, Ji SY, Kim DH, Kim MY, et al. Phloroglucinol inhibits oxidative-stress-induced cytotoxicity in C2C12 murine myoblasts through NRF-2-mediated activation of HO-1. Int J Mol Sci. 2023;24(5):4637. https://doi.org/10.3390/ijms24054637
  7. 7. Khan F, Tabassum N, Bamunuarachchi NI, Kim Y. Phloroglucinol and its derivatives: antimicrobial properties toward microbial pathogens. J Agric Food Chem. 2022;70(16):4817-38. https://doi.org/10.1021/acs.jafc.2c00532
  8. 8. Xie Y, Lu J, Yang T, Chen C, Bao Y, Jiang L, et al. Phloroglucinol, a clinical-used antispasmodic, inhibits amyloid aggregation and degrades the pre-formed amyloid proteins. Int J Biol Macromol. 2022;213:675-89. https://doi.org/10.1016/j.ijbiomac.2022.06.008
  9. 9. Yang E-J, Mahmood U, Kim H, Choi M, Choi Y, Lee J-P, et al. Phloroglucinol ameliorates cognitive impairments by reducing the amyloid β peptide burden and pro-inflammatory cytokines in the hippocampus of 5XFAD mice. Free Radic Biol Med. 2018;126:221-34. https://doi.org/10.1016/j.freeradbiomed.2018.08.016
  10. 10. Boubaker H, Boukef R, Claessens Y, Bouida W, Grissa MH, Beltaief K, et al. Phloroglucinol as an adjuvant analgesic to treat renal colic. Am J Emerg Med. 2010;28(6):720-3. https://doi.org/10.1016/j.ajem.2009.04.030
  11. 11. Weiss S, Waidha K, Rajendran S, Benharroch D, Khalilia J, Levy H, et al. In vitro and in vivo therapeutic potential of 6,6'-dihydroxythiobinupharidine (DTBN) from Nuphar lutea on cells and K18-hACE2 mice infected with SARS-CoV-2. Int J Mol Sci. 2023;24(9):8327. https://doi.org/10.3390/ijms24098327
  12. 12. Dalvie ED, Gopas J, Golan-Goldhirsh A, Osheroff N. 6,6′-Dihydroxythiobinupharidine as a poison of human type II topoisomerases. Bioorg Med Chem Lett. 2019;29(15):1881-5. https://doi.org/10.1016/j.bmcl.2019.06.003
  13. 13. Landau D, Khalilia J, Arazi E, Tobar AF, Benharroch D, Golan-Goldhrish A, et al. A Nuphar lutea plant active ingredient, 6,6′-dihydroxythiobinupharidine, ameliorates kidney damage and inflammation in a mouse model of chronic kidney disease. Sci Rep. 2024;14:7577. https://doi.org/10.1038/s41598-024-58055-1
  14. 14. Waidha K, Anto NP, Jayaram DR, Golan-Goldhirsh A, Rajendran S, Livneh E, et al. 6,6'-Dihydroxythiobinupharidine (DTBN) purified from Nuphar lutea leaves is an inhibitor of protein kinase C catalytic activity. Molecules. 2021;26(9):2785. https://doi.org/10.3390/molecules26092785
  15. 15. Singh N, Nandy SK, Jyoti A, Saxena J, Sharma A, Siddiqui AJ, et al. Protein Kinase C (PKC) in neurological health: implications for Alzheimer's disease and chronic alcohol consumption. Brain Sci. 2024;14(6):554. https://doi.org/10.3390/brainsci14060554
  16. 16. Gong C-X, Dai C-L, Liu F, Iqbal K. Multi-Targets: An unconventional drug development strategy for Alzheimer’s Disease. Front Aging Neurosci. 2022;14:837649. https://doi.org/10.3389/fnagi.2022.837649
  17. 17. Dighe S, Jog S, Momin M, Sawarkar S, Omri A. Intranasal drug delivery by nanotechnology: advances in and challenges for Alzheimer's disease management. Pharmaceutics. 2023;16(1):58. https://doi.org/10.3390/pharmaceutics16010058
  18. 18. Kaur A, Nigam K, Bhatnagar I, Sukhpal H, Awasthy S, Shankar S, et al. Treatment of Alzheimer's diseases using donepezil nanoemulsion: an intranasal approach. Drug Deliv Transl Res. 2020;10(6):1862-75. https://doi.org/10.1007/s13346-020-00754-z
  19. 19. Song Y, Wang X, Wang X, Wang J, Hao Q, Hao J, et al. Osthole-loaded nanoemulsion enhances brain target in the treatment of Alzheimer's disease via intranasal administration. Oxid Med Cell Longev. 2021;2021:8844455. https://doi.org/10.1155/2021/8844455
  20. 20. Ting P, Srinuanchai W, Suttisansanee U, Tuntipopipat S, Charoenkiatkul S, Praengam K, et al. Development of chrysin loaded oil-in-water nanoemulsion for improving bioaccessibility. Foods. 2021;10(8):1912. https://doi.org/10.3390/foods10081912
  21. 21. Kannavou M, Karali K, Katsila T, Siapi E, Marazioti A, Klepetsanis P, et al. Development and comparative in vitro and in vivo study of BNN27 mucoadhesive liposomes and nanoemulsions for nose-to-brain delivery. Pharmaceutics. 2023;15(2):419. https://doi.org/10.3390/pharmaceutics15020419
  22. 22. Jiang Y, Jiang Y, Ding Z, Yu Q. Investigation of the “Nose-to-Brain” pathways in intranasal HupA nanoemulsions and evaluation of their in vivo pharmacokinetics and brain-targeting ability. Int J Nanomedicine. 2022:17:3443-56. https://doi.org/10.2147/IJN.S369978
  23. 23. Shen X, Cui Z, Wei Y, Huo Y, Yu D, Zhang X, et al. Exploring the potential to enhance drug distribution in the brain subregion via intranasal delivery of nanoemulsion in combination with borneol as a guider. Asian J Pharm Sci. 2023;18(6):100778. https://doi.org/10.1016/j.ajps.2023.100778
  24. 24. Handa M, Sanap SN, Bhatta RS, Patil GP, Palkhade R, Singh DP, et al. Simultaneous intranasal codelivery of donepezil and memantine in a nanocolloidal carrier: optimization, pharmacokinetics, and pharmacodynamics studies. Mol Pharm. 2023;20(9):4714-28. https://doi.org/10.1021/acs.molpharmaceut.3c00454
  25. 25. Abdulla NA, Balata GF, El-Ghamry HA, Gomaa E. Intranasal delivery of Clozapine using nanoemulsion-based in-situ gels: an approach for bioavailability enhancement. Saudi Pharm J. 2021;29(12):1466-85. https://doi.org/10.1016/j.jsps.2021.11.006
  26. 26. Zhu Y, Chen T, Feng T, Zhang J, Meng Z, Zhang N, et al. Fabrication and biological activities of all-in-one composite nanoemulsion based on Blumea Balsamifera oil-tea tree oil. Molecules. 2023;28(15):5889. https://doi.org/10.3390/molecules28155889
  27. 27. de Barros AOdS, Pinto SR, dos Reis SRR, Ricci-junior E, Alencar LMR, Bellei NCJ, et al. Polymeric nanoparticles and nanomicelles of hydroxychloroquine co-loaded with azithromycin potentiate anti-SARS-CoV-2 effect. J Nanostruct Chem. 2023;13:263-81. https://doi.org/10.1007/s40097-022-00476-3
  28. 28. Khalid A, Arshad MU, Imran A, Haroon Khalid S, Shah MA. Development, stabilization, and characterization of nanoemulsion of vitamin D3-enriched canola oil. Front Nutr. 2023;10:1205200. https://doi.org/10.3389/fnut.2023.1205200
  29. 29. Gaber DA, Alsubaiyel AM, Alabdulrahim AK, Alharbi HZ, Aldubaikhy RM, Alharbi RS, et al. Nano-emulsion based gel for topical delivery of an anti-inflammatory drug: in vitro and in vivo evaluation. Drug Des Devel Ther. 2023;17:1435-51. https://doi.org/10.2147/DDDT.S407475
  30. 30. Krisanti EA, Kirana DP, Mulia K. Nanoemulsions containing Garcinia mangostana L. pericarp extract for topical applications: development, characterization, and in vitro percutaneous penetration assay. PLoS One. 2021;16(12):e0261792. https://doi.org/10.1371/journal.pone.0261792
  31. 31. Hashem AH, Doghish AS, Ismail A, Hassanin MMH, Okla MK, Saleh IA, et al. A novel nanoemulsion based on clove and thyme essential oils: characterization, antibacterial, antibiofilm and anticancer activities. Electron J Biotechnol. 2024;68:20-30. https://doi.org/10.1016/j.ejbt.2023.12.001
  32. 32. Ahmad N, Ahmad R, Al-Qudaihi A, Alaseel SE, Fita IZ, Khalid MS, et al. Preparation of a novel curcumin nanoemulsion by ultrasonication and its comparative effects in wound healing and the treatment of inflammation. RSC Adv. 2019;9:20192. https://doi.org/10.1039/c9ra03102b
  33. 33. Kotta S, Aldawsari HM, Badr-Eldin SM, Alhakamy NA, Shadab Md. Coconut oil-based resveratrol nanoemulsion: optimization using response surface methodology, stability assessment and pharmacokinetic evaluation. Food Chem. 2021;357:129721. https://doi.org/10.1016/j.foodchem.2021.129721
  34. 34. Patel MR, Patel MH, Patel RB. Preparation and in vitro/ex vivo evaluation of nanoemulsion for transnasal delivery of paliperidone. Appl Nanosci. 2016;6(8):1095-104. https://doi.org/10.1007/s13204-016-0527-x
  35. 35. Pavoni L, Perinelli DR, Bonacucina G, Cespi M, Palmieri GF. An overview of micro- and nanoemulsions as vehicles for essential oils: formulation, preparation and stability. Nanomaterials. 2020;10:135. https://doi.org/10.3390/nano10010135
  36. 36. Khani S, Keyhanfar F, Amani A. Design and evaluation of oral nanoemulsion drug delivery system of mebudipine. Drug Deliv. 2015;23(6):2035-43. https://doi.org/10.3109/10717544.2015.1088597
  37. 37. Elbardisy B, Galal S, Abdelmonsif DA, Boraie N. Intranasal Tadalafil nanoemulsions: formulation, characterization and pharmacodynamic evaluation. Pharm Dev Technol. 2019;24(9):1083-94. https://doi.org/10.1080/10837450.2019.1631846
  38. 38. Chong W-T, Tan C-P, Cheah Y-K, Lajis AMB, Dian NLHM, Kanagaratnam S, et al. Optimization of process parameters in preparation of tocotrienol-rich red palm oil-based nanoemulsion stabilized by Tween80-Span 80 using response surface methodology. PLoS One. 2018;13(8):e0202771. https://doi.org/10.1371/journal.pone.0202771
  39. 39. Taher SS, Al-Kinani KK, Hammoudi ZM, Ghareeb MM. Co-surfactant effect of polyethylene glycol 400 on microemulsion using BCS class II model drug. J Adv Pharm Educ Res. 2022;12(1):63-9. https://doi.org/10.51847/1h17TZqgyI
  40. 40. Yuliani S, Noveriza R. Effect of carrier oil and co-solvent on the formation of clove oil nanoemulsion by phase inversion technique. IOP Conf Ser: Earth Environ Sci. 2019;309:012036. https://doi.org/10.1088/1755-1315/309/1/012036
  41. 41. Ferreira AC, Sullo A, Winston S, Norton IT, Norton-Welch AB. Influence of ethanol on emulsions stabilized by low molecular weight surfactants. J Food Sci. 2020;85(1):28-35. https://doi.org/10.1111/1750-3841.14947
  42. 42. Miksusanti, Apriani EF, Bihurinin AHB. Optimization of tween 80 and PEG-400 concentration in Indonesian virgin coconut oil nanoemulsion as antibacterial against Staphylococcus aureus. Sains Malaysiana. 2023;52(4):1259-72. https://doi.org/10.17576/jsm-2023-5204-17
  43. 43. 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. https://doi.org/10.1016/j.ejpb.2006.10.014
  44. 44. Akhtar J, Siddiqui HH, Fareed S, Badruddeen Khalid M, Aqil M. Nanoemulsion: for improved oral delivery of repaglinide. Drug Deliv. 2016;23(6):2026-34. https://doi.org/10.3109/10717544.2015.1077290
  45. 45. Roohinejad S, Oey I, Wen J, Lee SJ, Everett DW, Burritt DJ. Formulation of oil-in-water β-carotene microemulsions: effect of oil type and fatty acid chain length. Food Chem. 2015;174:270-8. https://doi.org/10.1016/j.foodchem.2014.11.056
  46. 46. Balata GF, Essa EA, Shamardl HA. Zaidan SH, Abourehab MAS. Self-emulsifying drug delivery systems as a tool to improve solubility and bioavailability of resveratrol. Drug Des Dev Ther. 2016;10:117-28. https://doi.org/10.2147/DDDT.S95905
  47. 47. Hassan AK. Effective surfactants blend concentration determination for O/W emulsion stabilization by two nonionic surfactants by simple linear regression. Indian J Pharm Sci. 2015;77(4):461-9.
  48. 48. Partal P, Guerrero A, Berjano M, Gallegos C. Influence of concentration and temperature on the flow behavior of oil-in-water emulsions stabilized by sucrose palmitate. J Am Oil Chem Soc. 1997;74(10):1203-12. https://doi.org/10.1007/s11746-997-0046-8
  49. 49. Abdul Aziz ZA, Nasir HM, Ahmad A, Setapar SH, Ahmad H, Noor MHM, et al. Enrichment of eucalyptus oil nanoemulsion by micellar nanotechnology: transdermal analgesic activity using hot plate test in rats’ assay. Sci Rep. 2019;9:1-16. https://doi.org/10.1038/s41598-019-50134-y
  50. 50. Schreiner TB, Santamaria-Echart A, Ribeiro A, Peres AM, Dias MM, Pinho SP, et al. Formulation and optimization of nanoemulsions using the natural surfactant saponin from quillaja bark. Molecules. 2020;25:1538. https://doi.org/10.3390/molecules25071538

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