co2 assisted nano amorphization techniques nanosuspension preparation

1. Preparation Characterization and in
vivo evaluation of amorphous
tacrolimus nano suspensions produced
using CO2-assisted in situ nano
amorphization method
SUCHANDRA BAGCHI
M.S. PHARM (PHARMACEUTICS)
FIRST YEAR
NIPERA1517PE10

2. INTERNATIONAL JOURNAL OF
PHARMACEUTICS
ELSEVIER
3.56

3. FLOW OF PRESENTATION
INTRODUCTION
MATERIALS AND METHODS
CHARACTERIZATION OF NANOSUSPENSION
RESULTS AND DISCUSSION
CONCLUSION

4. INTRODUCTION
“Very finely colloid biphasic, dispersed and solid drug particles
in aqueous vehicle, size below 1µm without any matrix
material stabilized by surfactant and polymers and prepared
by suitable methods for drug delivery applications through
various routes of administration”

5. MATERIALS
 Tacrolimus- Huifengda Chemical Industrial co.ltd(Jinan,China)
 Prograf - Astellas Ireland Co. ltd.(Ireland)
 Poloxamer 407 and 188 –BASF
 D-alpha-tocopheryl polyethylene glycol 1000 succinate –Aladdin reagent
 Hydroxy propyl methylcellulose – Anhui Shanhe Pharmaceutical Excipients
 Citrazinic acid, sodium hydrogen carbonate and ammonium phosphate
monobasic- Tiajin Bodi
 Polyethylene glycol 6000- Sinopharm Chemical Reagent
 Ethanol –Tiajin damao
 Acetonitrile, dichloromethane and n-hexane- Concord Technology

6. COMPOSITION
 FORMULATION TABLE for FK506 NANOSUSPENSIONS
PHASE COMPONE
NTS %
(w/w)
FK506-
TPGS
FK506-F68 FK506-
F127
FK506-
HPMC
Organic
acid phase
FK506 1 1 1 1
TPGS 1 - - -
F68 - 1 - -
F127 - - 1 -
HPMC - - - 1
Citrazinic
acid
3 3 3 3
Carbonated
aqueous
phase
Sodium
hydrogen
carbonate
3.6 3.6 3.6 3.6

7. PREPARATION OF TACROLIMUS
NANOSUSPENSION

8. CHARACTERIZATION OF
NANOSUSPENSIONS
 PARTICLE SIZE ANALYSIS OF FK506 NANOSUSPENSIONS
FORMULATION PARTICLE SIZE PDI
FK506/TPGS 348.2±16.7 0.238±0.004
FK506/F68 402.7±17.9 0.127±0.086
FK506/F127 302.8±2.0 0.158±0.098
FK506/HPMC - -

9. LYOPHILIZATION
To check physicochemical properties of the aqueous nano suspension (FDU-1100)
•FK506-NA + 4%(w/v) PEG6000
•FK506-NB+ 6%(w/v) PEG6000
•FK506-NC+ 3%(w/v)PEG6000
The samples were frozen in a refrigerator at -80 ºC for 12 hours then transferred
to FDU-1100
Subsequently temperature was maintained at -40 ºC for 3 hours
Then the samples were lyophilized at the temperature of -20 ºc for 15 minutes
At the end of freeze drying stage the chamber temperature was holding at 30 ºC
for 3hours.

10. CHARACTERIZATION

11. CHARACTERIZATION
 PARTICLE SIZE OF FK506 NANOSUSPENSION
PREPARATION BEFORE
LYOPHILIZATION
AFTER LYOPHILIZATION
FK506-NA 167.3±2.8 180±10.1
FK506-NB 302.8±2.0 339.1±15.02
FK506-NC 513.5±15.1 491.1±14.71

12. MORPHOLOGY
 FK506-NA FK506-NB FK506-NC
 BY TEM NA and NC were having spherical shape whereas NB was having worm like
due to F127 amphiphilic triblock copolymer consisted of PEO(polyethylene oxide)
and polypropylene oxide (PPO)
 Hydrophilic PEO chains outer layer of nanoparticles stretching to water phase and
Hydrophobic PPO chain might fix on the surface particles

13. CRYSTALLINE STATE
DSC PROFILE
DSC curves of coarse
FK506 (a),
blank excipients(b),
physical mixtures (c),
FK506-NA (d),
FK506-NB (e) and
FK506-NC (f).

14. XRPD
X-ray diffraction results of
coarse FK506 (a),
blank excipients (b),
physicalmixture (c),
FK506-NA (d),
FK506-NB (e) and
FK506-NC (f).

15. STABILITY STUDIES
Diameter and Polydispersity index of FK506-NA (A),
FK506-NB (B) and
FK506-NC (C) at different time .The data are mean ± SD (n=3).

16. IN VITRO DISSOLUTION STUDIES
Dissolution profiles of
Prograf®, FK506-NA,
FK506-NB, FK506-NC and
FK506-D.

17. PHARMACOKINETIC STUDIES
Whole blood
concentration-time
profiles of FK506 after
oral administration of
five different
formulations in rats (n
= 5).

18. RESULT
PARAMETERS PROGRAF FK506-NA FK506-NB FK506-NC FK506-D
Cmax(µg/ml) 385.57±122.
20
666.43±17
3.14
789.98±94.7
9
565.19±146.
51
205.62±94.75
Tmax(min) 150±34.64 72±61.30 52.5±8.66 32±12.55 69±63.29
T ½(min) 136.07±28.2
8
109.69±21.
30
97.06±14.76 269.44±130.
15
163.68±55.17
AUC 0-12 83896.6±224
26.6
115919.5±
20251.9
125597.5±20
839.3
99523.0±179
89.9
40490.6±20689.8
F% 100 138 150 119 48
The pharmacokinetic parameters of FK506 after oral
administration of Prograf®, FK506-NA, FK506-NB, FK506-
NC and FK506-D
to rats (n = 5).

19. CONCLUSION
In this study FK506 nanosuspensions with different particle sizes were prepared
using in situ nanoamorphization method which was proved to be simple,
effective and robust technique for amorphous nanosuspensions of poorly water
soluble drugs. Physical characterizations illustrated that the FK506
nanosuspensions were in amorphous state.
From the in vitro dissolution and pharmacokinetic study these results indicated
that the dissolution rate and the bioavailability were increased as surface area
augmenting.
In conclusion the in situ amorphization method is valid technique to produce
amorphous FK506 nanosuspensions and has great potential for increasing
bioavailability