Spherical Amorphous Nanoparticles from Birch Bark Triterpenoids: Study of the Conditions for the Formation

Triterpenoids from birch bark are valuable biologically active substances. They exhibit important biological properties. However, triterpenoids are insoluble in water due to their hydrophobicity. In order to increase their bioavailability we developed a technique for obtaining spherical amorphous nanoparticles (SANP) by their precipitation with water from a solution in THF. In the form of nanoparticles, triterpenoids show additional useful properties: they can be loaded with hydrophobic substances. SANP were also shown to be effective immunological adjuvants. This article is devoted to the study of conditions that could optimize their production. As a result, it was shown that among the three investigated solvents miscible with water - acetone, dioxane, THF -the latter gives the best results. Lowering temperature decreases the size of SANP. An increased temperature leads to the formation of betulin crystals. When the concentration of triterpenoids in THF is higher than 5 mg/ml, crystals of betulin also begin to form. The results of this study lead to the assumption that the main parameter of the process determining the formation of SANP is the rate of the solvent diffusion into water.

1. Sergeev G.B. Dimensional effects in nanochemistry. Rossiyskiy khimicheskiy zhurnal (Zhurnal Rosssiyskogo khimicheskogo obshchestva im. D. I. Mendeleyeva) [Russian Chemical Journal (Journal of the D.I. Mendeleev Russian Chemical Society)] 2002; XLVI(5):22-29. (in Russ.). URL: http://www.chem.msu.su/rus/jvho/2002-5/22.pdf

2. Sakurai Y, Kajimoto K., Hatakeyama H., Harashima H. Advances in an active and passive targeting to tumor and adipose tissues. Expert Opin. Drug Deliv. 2015; 12(1):41-52. https://doi.org/10.1517/17425247.2015.955847

3. Chang C., Meikle T.G., Su Y, Wang X., Dekiwadia C., Drummond C.J., Conn C.E., Yang Y. Encapsulation in egg white protein nanoparticles protects anti-oxidant activity of curcumin. Food Chem. 2019; 280:65-72. https://doi.org/10.1016/j.foodchem.2018.11.124

4. Stvolinsky S.L., Antonova N.A., Kulikova O.I., Lopachev A.V., Abaimov D.A., Al-Baidani I., Lopacheva O.M., Fedorova T.N., Kaplun A.P., Sorokoumova G.M. Lipoilcarnosine: Synthesis, study of physico-chemical and antioxidant properties, biological activity. Biomeditsinskya khimiya [Biomedical Chemistry]. 2018. 64(3):268-275. (in Russ.). URL: https://elibrary.ru/title_about.asp?id=25613

5. Ong H.J., Pinal R. Drug solubilization by means of a surface-modified edible biopolymer enabled by hot melt extrusion. J. Pharm. Sci. 2018; 107(1):402-411. https://doi.org/10.1016/j.xphs.2017.10.022

6. Mineeva M.F. Antihypoxic agent: patent RU 2003137066. Priority from 12/24/2003. (in Russ.)

7. YingMeei Tan, Rong Yu, Pezzuto J.M. Betulinic acid - induced programmed cell death in human melanoma cells involves mitogen-activated protein kinase activation. Clin. Cancer Res. 2003; 9:2866-2875. https://doi.org/10.1016/j.xphs.2017.10.022

8. Cichewicz R.H., Kouzi S.A. Chemistry, biological activity, and chemotherapeutic potential of betulinic acid for the prevention and treatment of cancer and HIV infection. Med. Res. Rev. 2004; 24(1):90-114. https://doi.org/10.1002/med.10053

9. Akkar A., Miller R. H. Solubilisation by emulsification. Novel formulation principle of poorly soluble drugs for intravenous administration. Pharm. Ind. 2004; 66(12): 1537-1544.

10. Pacheco L.F., Carmona-Ribeiro A.M. Effects of synthetic lipids on solubilization and colloid stability of hydrophobic drugs. J. Colloid Interface Sci. 2003; 258(1):146-154. https://doi.org/10.1016/S0021-9797(02)00103-0

11. Palma S., Manzo R.H., Allemandi D., Fratoni L., Lo Nostro P Solubilization of hydrophobic drugs in octanoyl-6-O-ascorbic acid micellar dispersions. J. Pharm. Sci. 2002; 91(8):1810-1816. https://doi.org/10.1002/jps.10180

12. Ismail A.A., Gouda M.W., Motawi M.M. Micellar solubilization of barbiturates. I. Solubilities of certain barbiturates in polysorbates of varying hydrophobic chain length. J. Pharm. Sci. 1970; 59(2):220-224. PMID: 5411344

13. Shahgaldian P., Gualbert J., Aissa K., Coleman A.W. A study of the freeze-drying conditions of calixarene based solid lipid nanoparticles. Eur. J. Pharm. Biopharm. 2003; 55(2) P. 181-184. https://doi.org/10.1016/S0939-6411(02)00196-0

14. Chistyakov A.N., Presnova G.A., Balakshin V.V., Kaplun A.P. The composition of biologically active substances and a method for its nanodispersion: patent RU 2322091. Publ. 04/04/2008. Bull. No. 11. (in Russ.)

15. Kaplun A.P., Bezrukov D.A., Popenko VI., Shvets VI. Spherical amorphous nanoparticles from birch bark triterpenoids - a novel type of submicronic vehicle for drug delivery. Biofarmatsevticheskiy zhurnal = Russian Journal of Biopharmaceuticals. 2011; 3(2):28-40. (in Russ.)

16. Gambaryan A.S., Boravlyova E.Y, Bikova N.V., Kaplun A.P. Comparative testing of subunit, adjuvanted whole-virion inactivated and live anti-influenza vaccines against homologous and heterosubtypic challenge. Meditsinskaya virusologiya [Medical Virology]. 2014; 28(1):37-58. (in Russ.)

17. Krasilnikov I.V, Kulish D.M., Brazhkin A.L., Doronin A.N. The method of obtaining an adjuvant for vaccines: patent RU 2545717. Publ. 04/10/2015. Bull. No. 10. (in Russ.)

18. Kaplun A.P., Pomogaev A.I., Lonina N.N., Dubovik E.G. Reduction of silver ions on the surface of spherical amorphous nanoparticles from birch bark triterpenoids. Unikal'nyye issledovaniya XXI veka [Unique studies of the XXI century]. 2015; 7(7): 148-152. (in Russ.). URL: http://elibrary.ru/item.asp?id=23942980

19. Boguslavsky L.I., Buslaeva T.M., Fomichev V.V., Kopylova E.V, Kaplun A.P., Popenko VI. Synthesis of BaSO4 nanoparticles in the water - tetrahydrofuran system. Russian Journal of Physical Chemistry. 2015; 89(2):256-260.

20. Khlebnikov VK., Boguslavsky L.I., Popenko V.I., Kaplun A.P., Shvets VI. Method of studying depth distribution of therapeutic and diagnostic substances inside spherical amorphous nanoparticles. Rossiyskiye nanotekhnologii = Nanotechnologies in Russia. 2010; 5(9-10):709-714. (in Russ.)

21. Schubert M.A., Muller-Goymann C.C. Solvent injection as a new approach for manufacturing lipid nanoparticles - evaluation of the method and process parameters. Eur. J. Pharm. Biopharm. 2002; 20:300-307. https://doi.org/10.1016/S0939-6411(02)00130-3