HAL Id: hal-02615965
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Submitted on 24 May 2020
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The same nucleoside crosslinking agent to develop hydrogel and organogel platforms
Marion Pitorre, Guillaume Bastiat, Le Pham, Jean-Pierre Benoît
To cite this version:
Marion Pitorre, Guillaume Bastiat, Le Pham, Jean-Pierre Benoît. The same nucleoside crosslinking agent to develop hydrogel and organogel platforms. 10th World Meeting on Pharmaceutics, Bio- pharmaceutics and Pharmaceutical Technology, Apr 2016, Glasgow, United Kingdom. 2016. �hal- 02615965�
The same nucleoside crosslinking agent to develop hydrogel and organogel platforms
Marion Pitorre
1,2, Guillaume Bastiat
1,2, Le Thuy Trang Pham
1,2and Jean-Pierre Benoit
1,21
LUNAM, MINT, Angers, France ;
2INSERM, U1066, Angers
• Gels are attractive pharmaceutical systems to develop local administration and/or drug sustained release therapy.
• Nanoparticle-loaded gels combine the gel advantages with the nanoparticle properties:
stealthiness, targeting site, decreased toxicity and protection of the encapsulated drug from degradation.
• Previously, the formulation of gemcitabine-loaded lipid nanocapsules (LNCs) formed an unexpected and spontaneous gelling [1].
• This hydrogel was due to H-bond interactions between gemcitabine moieties exposed at the LNC surface.
• The subcutaneous administration of this hydrogel in mice showed a sustained release of LNCs along with a progressive accumulation in lymph nodes, and allowed to combat mediastinal metastases issued from an orthotopic non-small-cell lung cancer model [2].
We decided to develop the same platform without the use of gemcitabine (highly toxic component), replacing it by an endogenous molecule with similar properties.
References
[1] E. Moysan et al, “An innovative hydrogel of gemcitabine- loaded lipid nanocapsules: when the drug is a key player of the nanomedicine structure,” Soft Matter, vol. 10, 1767–1777, 2014.
[2] N. Wauthoz et al, “Safe lipid nanocapsule-based gel technology to target lymph nodes and combat mediastinal metastases from an orthotopic non-small-cell lung cancer model in SCID-CB17 mice,” Nanomedicine Nanotechnol. Biol. Med., vol.
11, 1237–1245, 2015.
Acknowledgment
This work was financially supported by EuroNanoMed2
ERA-NET within the research program NICHE.
Contact
marion.pitorre@univ-angers.fr guillaume.bastiat@univ-angers.fr jean-pierre.benoit@univ-angers.fr
Conclusion
Cyt-C16 is a very promising crosslinking agent to obtain two gel platforms with opposite properties. The LNC-based hydrogel could allow sustained release of lipophilic drug loaded nanocarriers after subcutaneous administration. Whereas, sustained release of hydrophilic components could be obtained from the organogel.
Cyt-C16 was also able to form an organogel. The molecule was simply solubilized in an oil phase at high temperature and a gel was formed after cooling.
Viscoelastic properties of the organogel were enhanced with increasing Cyt-C16 concentration (Kinexus® rheometer, Malvern Instruments S.A,).
(mean ± sd; n=3)
• Rheological properties
A gel form could be obtained at a Cyt-C16 concentration of 1 or 2.5% (w/w Labrafac®) and LNC concentration higher than 0.2 g/mL, for LNC sizes from 50 to 100 nm. For all the gels, phase transition temperatures were higher than the body temperature (> 70°C) (Kinexus® rheometer, Malvern Instruments S.A). (mean ± sd; n=3)
• Cytotoxicity
Cell mortality study on a macrophage cell line (derived from THP-1 cell line) confirmed the safety of Cyt-C16, comparing LNCs with and without Cyt-C16.
(48h incubation, 37°C, MTT test, mean ± sd, n=3)
An administration with a syringe does not alter the gel state of the formulation. Inclusion of urea, sodium chloride and ethanol (60 mol for 1 mol Cyt-C16) in the formulation to inhibit physical association, did not change the rheological properties of the hydrogel.
1E+00 1E+01 1E+02 1E+03 1E+04 1E+05 1E+06
0 5 10
G' and G'' (Pa)
Cyt-C16 concentration (% w/w labrafac)
G' G''
Palmitoyl cytidine (Cyt-C16)
Cytidine is an endogenous
molecule and its structure is close to gemcitabine.
G’ : elastic modulus G’’ : viscous modulus
0,00 20,00 40,00 60,00 80,00
0,1 1 10
Cell mortality (%)
LNC concentration (mg/mL)
Blank LNC 50 nm Blank LNC 100 nm
LNC 50 nm Cyt-C16 2,5 % LNC 100 nm Cyt-C16 2,5%
Cyt-C16 loaded LNC :
cytidine moieties are exposed at the surface. LNCs were produced by the classical phase inversion process [1].
An hydrogel resulted from H-bond interactions between LNCs, forming a network in water similar to a tridimensional pearl necklace structure.
Organogel was due to
interactions between Cyt-C16, forming fiber network in oil.
Kolliphor® HS15 Span® 80
Cytidine-C16
Labrafac® WL 1349
• Release
The in vitro release of LNCs from hydrogel study was performed in artificial extracellular matrix (pH 7.4) at 37°C over 3 days. LNCs were labelled with nile red dye and followed by fluorescence.
(2.5% w/w Labrafac®, mean ± sd; n=3)
0,05 0,5 5 50 500
10 100 1000
G' and G'' (Pa)
LNC concentration (mg/mL)
Blank LNC 50 nm - G' Blank LNC 50 nm - G''
LNC 50 nm - Cyt-C16 2.5% - G' LNC 50 nm - Cyt-C16 2.5% - G''
0 10 20 30
0 20 40 60 80
% of LNCs in suspension
Time (hours)
LNC 50 nm Cyt-C16 2.5%
LNC 100 nm Cyt-C16 2.5%
