Skip to main content

Synthesis and Evaluation of Substituted Poly(organophosphazenes) as a Novel Nanocarrier System for Combined Antimalarial Therapy of Primaquine and Dihydroartemisinin

Abstract

Purpose

The synthesis and evaluation of novel biodegradable poly(organophosphazenes) (36) namely poly[bis-(2-propoxy)]phosphazene (3) poly[bis(4-acetamidophenoxy)]phosphazene (4)poly[bis(4-formylphenoxy)]phosphazene (5) poly[bis(4-ethoxycarbonylanilino)]phosphazene (6) bearing various hydrophilic and hydrophobic side groups for their application as nonocarrier system for antimalarial drug delivery is described.

Methods

The characterization of polymers was carried out by IR, 1H-NMR and 31P-NMR. The molecular weights of these novel polyphosphazenes were determined using size exclusion chromatography with a Waters 515 HPLC Pump and a Waters 2414 refractive index detector. The degradation behavior was studied by 200 mg pellets of polymers in phosphate buffers pH 5.5, 6.8 and 7.4 at 37°C. The degradation process was monitored by changes of mass as function of time and surface morphology of polymer pellets. The developed combined drugs nanoparticles formulations were evaluated for antimalarial potential in P. berghei infected mice.

Results

These polymers exhibited hydrolytic degradability, which can afford applications to a variety of drug delivery systems. On the basis of these results, the synthesized polymers were employed as nanocarriers for targeted drug delivery of primaquine and dihydroartemisinin. The promising in vitro release of both the drugs from nanoparticles formulations provided an alternative therapeutic combination therapy regimen for the treatment of drug resistant malaria. The nanoparticles formulations tested in resistant strain of P. berghei infected mice showed 100% antimalarial activity.

Conclusions

The developed nanocarrier system provides an alternative combination regimen for the treatment of resistant malaria.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Scheme I
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12

Abbreviations

1H-NMR:

Proton nuclear magnetic resonance

31P-NMR:

Phosphorus nuclear magnetic resonance

ALP:

Alkaline phosphatase

ANOVA:

Analysis of variance

CPCSEA:

Committee for the purpose of control and supervision on experiments on animals

D2O:

Deuterated water

DHA:

Dihydroartemisinin

DMSO:

Dimethyl sulphoxide

DSC:

Differential scanning calorimetry

EDTA:

Ethylene diamine tetraacetic acid

EE:

Entrapment efficiency

ELISA:

Enzyme linked immuno sorbent assay

HPLC:

High performance liquid chromatography

IAEC:

Institutional animal ethical committee

IR:

Infrared

MHz:

Mega hertz

MST:

Mean survival time

Mw :

Molecular weight

NIMR:

National institute of malaria research, New Delhi, India

NP:

Nanoparticles

o/w:

Oil/water

PBS:

Phosphate buffer saline

PEG:

Polyethyleneglycol

ppm:

parts per million

PQ:

Primaquine

RBCs:

Red blood cells

SEM:

Scanning electron microscopy

SGOT:

Serum glutamic oxaloacetic transaminase

SGPT:

Serum glutamic pyruvate transaminase

TEM:

Transmission electron microscopy

Tg :

Glass transition temperature

TGA:

Thermogravimetric analysis

THF:

Tetrahydrofuran

UV:

Ultraviolet

Wt:

Weight

References

  1. Allcock HR. Inorganic polymers. New York: Allyn and Bacon; 1995.

    Google Scholar 

  2. Potin PH, Jeager RD. Polyphosphazenes: synthesis, structure, properties, application. Eur Polym J. 1991;4(5):341–58.

    Article  Google Scholar 

  3. Allcock HR. Chemistry and applications of polyphosphazenes. Wiley: Hoboken; 2003.

    Google Scholar 

  4. Allcock HR. Recent developments in polyphosphazene materials science. Curr Op Solid St. 2006;10:231–40.

    CAS  Article  Google Scholar 

  5. Luten J, Steenbergen VMJ, Lok MC, Graaff DAM, Nostrum VCF, Talsma H, et al. Degradable PEG-folate coated poly(DMAEA-co-BA)phosphazene-based polyplexes exhibit receptor-specific gene expression. Eur J Pharm Sci. 2008;33:241–51.

    CAS  PubMed  Article  Google Scholar 

  6. Allcock HR, Steely LB, Singh A. Hydrophobic and superhydrophobic surfaces from polyphosphazenes. Polym Int. 2006;55:621–5.

    CAS  Article  Google Scholar 

  7. Sethuraman S, Nair LS, El-Amin S, Nguyen MT, Singh A, Krogman N, et al. Mechanical properties and osteocompatibility of novel biodegradable alanine based polyphosphazenes: Side group effects. Acta Biomater. 2010;6:1931–7.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  8. Ottenbrite RA, Albertsson AC, Scott G, Vert M, Feijen J, Albertsson A, Scott G, Chiellini E. Biodegradable Polymers and Plastics. 1992;73–92.

  9. Siepmanna J, Gpferichb A. Mathematical modeling of bioerodible, polymeric drug delivery system. Adv Drug Deliv Rev. 2001;48:229.

    Article  Google Scholar 

  10. Laurencin CT, Morris CD, Jacques HP, Schwartz ER, Keaton AR, Zou L. Osteoblast culture on bioerodible polymers: studies of initial cell adhesion and spread. Polym Adv Tech. 1992;3:359–64.

    CAS  Article  Google Scholar 

  11. Lee SM, Chun CJ, Heo JY, Song SC. Injectable and thermosensitive poly(organophosphazene) hydrogels for a 5-fluorouracil delivery. J Appl Polym Sci. 2009;113:3831–9.

    CAS  Article  Google Scholar 

  12. Kang GD, Heo JY, Jung SB, Song SC. Controlling the thermosensitive gelation properties of poly(organophosphazenes) by blending. Macromol Rapid Commun. 2005;26:1615–8.

    CAS  Article  Google Scholar 

  13. Sharma R, Rawal RK, Malhotra M, Sharma AK, Bhardwaj TR. Design, synthesis and ex-vivo release studies of colon-specific polyphosphazene–anticancer drug conjugates. Bioorg Med Chem. 2014;22:1104–14.

    CAS  PubMed  Article  Google Scholar 

  14. Singla N, Sharma R, Bhardwaj TR. Design, synthesis and in-vitro evaluation of polymer-linked prodrug of methotrexate for the targeted delivery to the colon. Lett Drug Des Discov. 2014;11:601–10.

    CAS  Article  Google Scholar 

  15. Murthy RSR, Sapariya B, Solanki A. Sustained release implants of chloroquine phosphate for posible use in chemoprophylaxix of malaria. Indian J Exp Biol. 2001;39:902–5.

    PubMed  Google Scholar 

  16. Aditya NP, Patankar S, Madhusudhan B, Murthy RSR, Souto EB. Arthemeter-loaded lipid nanoparticles produced by modified thin-film hydration: Pharmacokinetics, toxicological and in vivo anti-malarial activity. Eur J Pharm Sci. 2010;40:448–55.

    CAS  PubMed  Article  Google Scholar 

  17. Kumar S, Singh RK, Sharma R, Murthy RSR, Bhardwaj TR. Design, synthesis and evaluation of antimalarial potential of polyphosphazene linked combination therapy of primaquine and dihydroartemisinin. Eur J Pharm Sci. 2014. doi:10.1016/j.ejps.2014.09.023.

    Google Scholar 

  18. Sohn YS, Cho YH, Baek H, Jung OS. Synthesis and properties of low molecular weight polyphosphazenes. Macromolecules. 1995;28:7566–8.

    CAS  Article  Google Scholar 

  19. Gumusderelioglu M, Gur A. Synthesis, characterization: in vitro degradation and cytotoxicity of poly[bis(ethyl-4-aminobutyro)phosphazene]. React Funct Polym. 2002;52:71–80.

    CAS  Article  Google Scholar 

  20. Huang KJ, Zhu CH. The production and characteristics of solid lipid nanoparticles. Biomaterials. 2003;24:1781–5.

    PubMed  Article  Google Scholar 

  21. Peters W, Robinson BL. The chemotherapy of rodent malarial. Studies on puronaridine and other manich base antimalarials. Ann Trop Med Parasitol. 1992;86:455–65.

    CAS  PubMed  Google Scholar 

  22. Mengiste B, Makonnen E, Urga K. In vivo antimalarial activity of Dodonaea angustifolia seed extracts against Plasmodium berghei in mice model. MEJS. 2012;4:47–63.

    Google Scholar 

  23. Raina N, Goyal AK, Pillai CR, Rath G. Development and characterization of artemether loaded solid lipid nanoparticles. IJPER. 2013;47(2):123–8.

    Google Scholar 

  24. Burkersroda FV, Schedl L, Gopfericha A. Why degradable polymers undergo surface erosion or bulk erosion? Biomaterials. 2002;23:4221–31.

    Article  Google Scholar 

  25. Deng M, Nair LS, Nukavarapu SP, Kumbar SG, Jiang T, Weikel AL, et al. Porous structures: in situ porous structures: a unique polymer erosion mechanism in biodegradable dipeptide-based polyphosphazene and polyester blends producing matrices for regenerative engineering. Adv Funct Mater. 2010;20:2743.

    PubMed Central  PubMed  Article  Google Scholar 

  26. Lakshmi S, Katti DS, Laurencin CT. Biodegradable polyphosphazenes for drug delivery applications. Adv Drug Deliv Rev. 2003;55:467–82.

    CAS  PubMed  Article  Google Scholar 

  27. Reddy RC, Vathsala PG, Keshamouni VG. Curcumin for malaria therapy. Biochem Biophys Res Commun. 2005;326:472–4.

    CAS  PubMed  Article  Google Scholar 

  28. Matthew C, Amy C, Robyn TB. Inhibition of intestinal tumors by curcumin is associated with changes in the intestinal immune cell profile. J Surg Res. 2000;89:169–75.

    Article  Google Scholar 

  29. Golenser J, Domb A, Teomim D. The treatment of animal models of malaria with iron chelators by the use of a novel polymeric device for slow drug release. J Pharmacol Exp Ther. 1997;281:1127–35.

    CAS  PubMed  Google Scholar 

  30. Crommelin DJA, Eling WMC, Steerenberg PA. Liposome and immunoliposome for control release or site specific delivery of anti-parasitic drugs and cytostatics. J Control Release. 1991;161:47–154.

    Google Scholar 

  31. Aditya NP, Chimote G, Gunalan K, Banerjee R, Patankar S, Madhusudhan B. Curcuminoids-loaded liposomes in combination with arteether protects against Plasmodium berghei infection in mice. Exp Parasitol. 2012;131:292–9. doi:10.1016/j.exppara.2012.04.010.

    CAS  PubMed  Article  Google Scholar 

Download references

ACKNOWLEDGMENTS AND DISCLOSURES

The authors acknowledge the financial support received from Life Science Research Board (LSRB) of Defense Research and Development Organization (DRDO), New Delhi (India) DL/81/48222/LSRB-232/SH & DD/2011. We are also thankful to Sh. Parveen Garg, Chairman, I.S.F. College of Pharmacy, Moga (Punjab) (India) for providing the necessary facilities to carry out the research work. We also acknowledge Punjab Technical University, Jalandhar (Punjab) (India) for providing necessary facilities for research work.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to T. R. Bhardwaj.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 2601 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kumar, S., Singh, R.K., Murthy, R.S.R. et al. Synthesis and Evaluation of Substituted Poly(organophosphazenes) as a Novel Nanocarrier System for Combined Antimalarial Therapy of Primaquine and Dihydroartemisinin. Pharm Res 32, 2736–2752 (2015). https://doi.org/10.1007/s11095-015-1659-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-015-1659-5

KEY WORDS

  • biodegradable polyphosphazenes
  • degradation
  • dihydroartemisinin
  • drug-resistant malaria
  • nanoparticles
  • primaquine