HAL Id: inserm-01375596
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Amorphous calcium carbonate based-microparticles for
peptide pulmonary delivery
Frederic Tewes, Oliviero Gobbo, Carsten Ehrhardt, Anne Healy
To cite this version:
Frederic Tewes, Oliviero Gobbo, Carsten Ehrhardt, Anne Healy. Amorphous calcium carbonate based-microparticles for peptide pulmonary delivery. ACS Applied Materials & Interfaces, Washington, D.C. : American Chemical Society, 2015, 8 (2), pp.1164-1175. �10.1021/acsami.5b09023�. �inserm-01375596�
Supporting Information
Amorphous calcium carbonate based-microparticles for peptide
pulmonary delivery
Frederic Tewes1, 2*, Oliviero L. Gobbo1, Carsten Ehrhardt1 and Anne Marie Healy1*
1: School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland
2: INSERM U 1070, Pôle Biologie-Santé, Université de Poitiers, Faculté de Médecine & Pharmacie, 1 rue Georges Bonnet, 86022 Poitiers Cedex, France
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Figure S1: Spray drying process scheme.Spray drying: The microparticles were manufactured in a one-step procedure using a Mini Spray Dryer B-290 (Büchi, Flawil, Switzerland). Two solutions were prepared separately and mixed during the process in a Y shape tubing connector fed to the spray dryer. One solution was composed of salmon calcitonin acetate salt (sCT) (0, 0.2 or 0.4 g L-1, Polypeptide Laboratories, Sweden) Ca(OH)2 (0.8 g L-1), hyaluronic acid sodium salt from Streptococcus equii (0.1 –
0.6 g L-1, Sigma-Aldrich, Dublin, Ireland) and 0.1% v/v of formic acid. The other solution was composed of ammonium carbonate ((NH4)2CO3)(1.2 – 6 g L-1) and alpha
1-antitrypsin (AAT) (0 or 0.2 g L-1). sCT and AAT were in separate solutions as their mixing induced the formation of a precipitate. The spray dryer was operated in an open-cycle suctionmode as follows: A 2-fluid nozzle was used to disperse the solution in the same direction as the flow of hot drying air (flowing at 630 L min-1); spraying air nozzle flow rate was 15 L min-1; inlet temperature ranged from 100 to 140 °C; feeding peristaltic pump was set at 30 % for the 2 solutions. These conditions resulted in an outlet temperature ranging from 47 to 70 °C.
S-2
Figure S2: FTIR spectra of Ca/HA composite microparticlesA) Formulated at different concentrations of HA, B) Loaded with sCT and AAT.
900 1100 1300 1500 1700 Norma liz ed ab so rbance Wavenumber (cm-1) HA-: 0.6 g/L HA-: 0.4 g/L HA-: 0.2 g/L HA-: 0.1 g/L 1592 1495 1421 1387 1368 1050 1081 1158 957 700 800 900 1000 1100 1200 Wavenumber (cm-1) HA-: 0.6 g/L HA-: 0.4 g/L HA-: 0.2 g/L HA-: 0.1 g/L 953 874 866 1047 1081 1155 793 774 900 1100 1300 1500 1700 No rmaliz ed ab sorba nce Wavenumber (cm-1) blank particles sCT CaCO3 sCT-AAT CaCO3 1476 1359 1050 1081 1158 1408 1591 1486 1361 1373 1385 1205 700 800 900 1000 1100 1200 Wavenumber (cm-1) Blank particles sCT-CaCO3 sCT AAT CaCO3 866 877 1050 1081 1158 792 777 777 724 1145
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Figure S3: TGA thermograms of Ca/HA composite particles formulated at different spray drying inlet temperatures (Tinlet).Thermogravimetric Analysis (TGA) was
performed using a Mettler TG 50 (Mettler Toledo, Greifensee, Switzerland). Samples were loaded into 40 µL aluminium open pans and run at a heating rate of 10 ºC min-1 from 25 – 420 ºC.
Thermogravimetric analysis of the particles formulated at various temperatures showed a 3-step decrease in mass with the increase in temperature on each thermogram. The first 3-step of 12 wt.% occurred at a temperature in the TGA lower than 200 °C and is assumed tobe related to water release from ACC. The second step of 10 wt.% had an onset temperature of 230 °Cand maybe due to the decomposition of HA. The third step of 10 wt.% had an onset temperature at 360-380 °C and corresponds to the melting/decomposition temperature of calcium formate.
0
200
400
Mass c
h
ang
e
(%)
Temperature (
o
C)
20%
120 °C
110 °C
100°C
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Figure S4: SEM micrographs of composite particles formulated withdifferent
concentrations of(NH4)2CO3. Carb1 (1.2 g L-1), Carb2 (2.4 g L-1), Carb3 (4.8 g L-1),
Carb4 (6 g L-1).
Carb2
Carb3
Carb4
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Figure S5: SEM micrographs of Ca/HA composite particles formulated withdifferent
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0 10 20 30 40 50 60 70 80 90 100 0 5 10 15 20 25 30 35 40 0 200 400 600 800 1000 1200 1400 1600 % P/P o Chang e In Mas s ( % ) Time (min)sCT-AAT CaCO3 particle % P/Po 15 25 35 45 55 In tensit y a.u. 2 qdegree sCT CaCO3 after DVS sCT CaCO3 Before DVS sCT-AAT CaCO3 after DVS sCT-AAT CaCO3 before DVS
F 26,3 F 31,3 F 16,0 F 41,1 V 27,1 F 28 V 24,9 V 32,8 F 49,8V 46,5
Figure S6: A) sCT-AAT Ca/HA composite particle water sorption versus time profiles
obtained at different % relative humidity, B) sCT-AAT Ca/HA particle water
sorption-desorption isotherm, C) XRD patterns of sCT-AAT and sCT Ca/HA particles after DVS
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Figure S7: sCT-AAT-loaded microparticles’aerodynamic diameter size distribution.
The aerodynamic diameter (AD) distribution of the particles was measured using an
Andersen cascade impactor (ACI). The flow rate was adjusted to 60 L min-1 in the powder
inhaler (Handihaler®, BoehringerIngelheim) and the time of aspiration was adjusted to obtain
4 L. The inhaler was filled with gelatin n°3 capsule loaded with 20 ± 2 mg of powder (n = 3). After inhaler actuation, particle deposition on the ACI was determined by calcium assay. The amount of particles with AD≤ 5.0 μm, expressed as a percentage of the emitted recovered dose, was considered as the fine particle fraction (FPF). The mass median aerodynamic diameter (MMAD) and FPF were calculated as previously described. (Tewes, F.; Gobbo, O. L.; Amaro, M. I.; Tajber, L.; Corrigan, O. I.; Ehrhardt, C.; Healy, A. M., Evaluation of HPβCD-PEG microparticles for salmon calcitonin administration via pulmonary delivery.
