Multiple-unit tablet of probiotic bacteria for improved storage stability, acid tolerability, and in vivo intestinal protective effect

Figures & data

Table 1 Composition of the probiotics-loaded ECPs-T formulation

Excipients	F1	F2	F3	F4	F5	F6	F7	F8	F9
ECPs	335	335	335	335	335	335	335	335	335
L-HPC 11	22.5	22.5	22.5	22.5	22.5	22.5	22.5	22.5	22.5
Crospovidone	5	5	5	5	5	5	5	5	5
HPC-EXF	20	20	20	20	20	20	20	20	20
Avicel PH102	37.5	–	–	–	–	–	37.5	37.5	37.5
Lactose monohydrate	–	37.5	–	–	–	–	–	–	–
Corn starch	–	–	37.5	–	–	–	–	–	–
Avicel PH105	–	–	–	37.5	–	–	–	–	–
Avicel PH101	–	–	–	–	37.5	–	–	–	–
Avicel PH200	–	–	–	–	–	37.5	–	–	–
Calcium silicate	–	–	–	–	–	–	15	25	35
Magnesium stearate	15	15	15	15	15	15	15	15	15

Note: Unit of measurement is mg; “–” indicates not added or not included in the formula.

Abbreviations: ECPs-T, enteric-coated pellets embedded tablet; ECPs, enteric-coated pellets; F, formulation; HPC, hydroxypropylcellulose; L-HPC, low-substituted hydroxypropyl cellulose.

Figure 1 Effect of cushioning agents on (A) flow properties of ECP powders; CI and angle of repose (indicated by the red circles), and (B) tablet hardness and disintegration time (indicated by the black circles) in distilled water.

Note: Data are expressed as mean ± SD values (n=3).
Abbreviations: CI, compressibility index; ECP, enteric-coated pellet; SD, standard deviation; F, formulation.

Figure 2 Effect of particle sizes of MCC granules on (A) flow properties of ECP powders; CI and angle of repose (indicated by the red circles), and (B) tablet hardness and disintegration time (indicated by the black circles) in distilled water.

Note: Data are expressed as mean ± SD values (n=3).
Abbreviations: CI, compressibility index; ECP, enteric-coated pellet; MCC, microcrystalline cellulose; SD, standard deviation; F, formulation.

Figure 3 Effect of calcium silicate combined with MCC on (A) flow properties of ECP powders; CI and angle of repose (indicated by the red circles), and (B) tablet hardness and disintegration time (indicated by the black circles) in distilled water.

Note: Data are expressed as mean ± SD values (n=3).
Abbreviations: CI, compressibility index; ECP, enteric-coated pellet; MCC, microcrystalline cellulose; SD, standard deviation; F, formulation.

Figure 4 Scanning electron microphotographs of (A) the surface of ECPs and (B) cross-section of ECPs-T.

Note: Arrow indicates the ECP embedded in the tablet.
Abbreviations: ECP, enteric-coated pellet; ECPs-T, ECPs embedded tablet.

Figure 5 Survival rate of probiotic bacteria during the dry powder coating process and under compression of ECPs into tablet dosage form.

Note: Data are expressed as mean ± SD values (n=3).
Abbreviations: ECPs, enteric-coated pellets; E. faecalis, Enterococcus faecalis; L. acidophilus, Lactobacillus acidophilus; SD, standard deviation.

Figure 6 Storage stability of (A) L. acidophilus and (B) E. faecalis in ECPs-T in comparison to corresponding bare bacteria, uncoated probiotic tablet, marketed products, and ECPs under ambient storage conditions (25°C/60% relative humidity) for 6 months.

Notes: To compare the bacterial storage viability, two marketed products, Andilac-S capsule (Il-Yang Pharm. Co., Ltd., Gyeonggi-do, Korea) or BIOR tablet (Unimed Pharmaceutical Inc., Seoul, Korea) containing L. acidophilus or E. faecalis, respectively, were used. Data represent the mean ± SD (n=3), and statistical analysis was performed using Student’s t-test; *P<0.05 versus bare bacteria; **P<0.05 versus ECPs.
Abbreviations: ECPs, enteric-coated pellets; ECPs-T, enteric-coated pellets embedded tablet; E. faecalis, Enterococcus faecalis; L. acidophilus, Lactobacillus acidophilus; SD, standard deviation.

Figure 7 Bacterial viability of (A) L. acidophilus and (B) E. faecalis inside ECPs-T after immersion in acidic medium in comparison to that of free bacteria, uncoated probiotic tablet, marketed products, and ECPs.

Notes: Data represent the mean ± SD (n=3), and statistical analysis was performed using Student’s t-test; *P<0.05 versus bare bacteria; **P<0.05 versus ECPs.
Abbreviations: ECPs, enteric-coated pellets; ECPs-T, enteric-coated pellets embedded tablet; E. faecalis, Enterococcus faecalis; L. acidophilus, Lactobacillus acidophilus.

Figure 8 Effect of repeated administration of ECPs-T formulation on plasma endotoxin concentrations in normal rats.

Notes: Data represent the mean ± SD (n=5), and statistical analysis was performed using Student’s t-test; *P<0.05 versus vehicle-treated group; **P<0.05 versus bare bacteria; and ***P<0.05 versus VSL#3 (Danisco, Madison, WI, USA). Plasma endotoxin levels were measured by LAL assay.
Abbreviations: ECPs-T, enteric-coated pellets embedded tablet; LAL, Limulus amebocyte lysate.