Copolymer Microgels for Oral

Delivery of Therapeutic Proteins

Table of Contents
Introduction
 Therapeutic proteins and use of Microgels for their
oral delivery
Objectives
Microgels
 Synthesis and structure
 Characterization of dry particles of microgel
 Microgels Compositions
 Microgels in Aqueous Conditions
Microgels
 Loading Capacity
 Protein Release Profiles
Conclusions
Perspectives
Acknowledgement
Introduction

Therapeutic Proteins
 Marketed Drugs based on Erythropoietin (Epo),
Interferon and Interleukin-2
 Advantages:
 High selectivity
 Lower risks of side effects
 Low dosage
 Complication:
 Administration by injection
Introduction

Microgels for Oral Delivery of Therapeutic

Proteins
Microgels:
 pH and temperature sensitive1
 protect API from proteolytic enzymes and low pH of
gastric fluids
 possess mucoadhesive properties2
 provide prolonged release of API at neutral pH

1
Bromberg L. et al., Langmuir, 2002
2
Bromberg L. et al., Polym. Prep., 1997
General Objectives
To synthesize and study copolymer microgels,
which potentially may be used for oral delivery
of therapeutic proteins
Pluronic®

( O ) PEO-PPO-PEO
( O ) ( O )y x H
x MW 1kD – 13kD
OH

x = 0 - 300
y = 15 - 80
x y x
Synthesis: block copolymers

70%
PEO content

20%

0%

Pluronic® F127 Pluronic® L92 Poly(propylene

glycol)
Synthesis: solution A
Synthesis
Synthesis: microgel structure

Bromberg L. et al., Langmuir, 2002

Bromberg L., Ind. Eng. Chem. Res, 2001
Size Distribution

90.00%
80.00%
70.00%
60.00%
% Weight

POP microgel
50.00%
L92 microgel
40.00%
F127 microgel
30.00%
20.00%
10.00%
0.00%
0.86 0.38 0.14 0.074
Mesh Opening mm
Microscopy
Microgel F127

Microgel POP

Microgel L92
 Flowability :Carr’s compressibility index

Lactose
37%

42%
Carbopol 1

41%
Mg POP

17%
Mg L92

28%
Mg F127

Lower Carr’s indices of compressibility are related to

less cohesiveness and greater fluidity2
1
Carbopol= Carbopol 971 NF (Noveon)
2
Carr RL. Evaluating flow properties of solids. Chem Eng. 1965
Degree of bonding
 Microgel wash-outs from all the washing steps
for each batch were evaporated and analyzed by
FTIR and HPLC to quantitate unbound
copolymers and acrylic acid monomers
 Dry microgel particles for each batch were
analysed by DSC to determine the efficacy of
washing
Content of Pluronic
Degree of Pluronic Bonding

98.00%
97.00% 96%
96.00%
95.00%
94.00%
93%
DoB %

93.00%
92%
92.00%
91.00%
90.00%
89.00%
88.00%
87.00%
POP microgel L92 microgel F127 microgel
Content of Poly(acrylic Acid)
No acrylic acid residues were
detected in the wash-outs by
either HPLC or FTIR analysis
Microgels Compositions
F127 Microgel L92 microgel

PEO
PEO 7%
25%
PPO
28%
PPO
Acrylic Acid Acrylic Acid
10%
65% 65%

POP microgel

PPO
36%

Acrylic Acid
64%
Microgels Swelling
20
18
16
14
12 pop
10 L92
S

8 F127

6
4
2
0
0 5 10 15
pH
Formulation

 Microgels were blended with 2%,10%,20% and

50% w/w of protein (BSA) and 0.5% w/w of
Magnesium Stearate was added as lubricant
 300mg tablets were compressed with 5000 Kgs
force applied
Experiment
 Release was evaluated at 37ºC for 48 hours
under simulated GI tract conditions
 First two hours – pH 2.2 fluid, low shaking
 Next 46 hours – phosphate buffer pH 6.8, vigorous
shaking
 BSA release was evaluated with the Bio-Rad RC
Protein Assay
Microgel F127 - Effect of Loading on Release
BSA release from Pluronic F127 Microgel

100.00%

90.00%

80.00%

70.00%

60.00%
BSA release (%)

50.00%

40.00%
30.00%
20.00%
10.00%
48
0.00% 24
8
50% BSA
5
4
3
20% BSA
2.5 pH2.2
10% BSA
2
2% BSA 1 pH6.8
0
Microgel L92 - Effect of Loading on Release
BSA release from L92 M icrogel

100.00%

90.00%

80.00%

70.00%
BSA release(%)

60.00%

50.00%

40.00%

30.00%

20.00%

10.00%
48
0.00% 24
8
5
pH2.2 50% BSA 4
)
3 e( H
20% BSA
2.5 Ti m
10 % BSA 2
pH6.8 2% BSA
1
0
Microgel POP - Effect of Loading on Release
BSA release from POP Microgel

100.00%

90.00%

80.00%

70.00%
BSA release (%)

60.00%

50.00%

40.00%

30.00%

20.00%

10.00%
48
0.00% 24
8
5
50% 4
)
BSA 20%
3
e (H
Ti m
pH2.2
2.5
BSA 10% 2
1
BSA 2%
pH6.8
0

BSA
Protein Release from Microgel Tablets

100.00
90.00
80.00
70.00

BSA release (%)

60.00
50.00
40.00
30.00
20.00
10.00
0.00
48
8 24
Lactose

5
Microgel F127

3 4
Microgel L92

2
Microgel POP

0 1
Carbopol

)
time (h

pH2.2

pH6.8
Conclusions
 Microgels L92 and F127, directly compressed
with 10% w/w loading of protein, protect the
API from low pH fluids for 2 hours
 These microgels provide 7 hour release of
proteins under simulated GI tract conditions
with a total release up to 98% of loaded
Perspectives
 Microgels L92 and F127 loaded with number of
therapeutic proteins and peptides are to be
tested in vivo
 Synthesis and testing of microgels, based on
other poloxamers, is in progress
Acknowledgment
 Faculté de pharmacie, Université de Montréal:
 Jean Norbert McMullen, Ph.D.
 Hildgen Patrice, Ph.D.
 Nekka Fahima, Ph.D.
 Supratek Pharma Inc.:
 Grzegorz Pietrzynski Ph.D
 Valery Alakhov, Ph.D.
 Kishore Patel, Ph.D.
 Tomasz Popek, Ph. D.
 Massachusetts Institute of Technology:
 Lev Bromberg, Ph. D.