GB CC Sirius 2015
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Transcript of GB CC Sirius 2015
1
New In Vitro Methods for Bio-Relevant Analysis of Both Small Molecules and Proteins
George ButcherTechnical Product Manager, Sirius Analytical
Forest Row, [email protected]
An introduction to Sirius Sirius was founded in 1989. We are a manufacturer and vendor of
instrumentation for measurement of physicochemical parameters. We also provide an Analytical Service, and measure thousands of
samples for hundreds of customers, worldwide, each year. In the US, we are based in Beverly, MA.
www.sirius-analytical.com
My presentation today:
- The need for improved in-vitro testing models
- Sirius inForm – dynamic dissolution testing instrument
- Introducing Sirius Scissor – a new instrument for biotherapeutics
- Example studies from both systems
4
GI Conditions & traditional dissolution
5
Historically:
• Develop pharmaceutical products to be manufactured in bulk
• Test criteria based on reproducibility of manufacture
The Rise of Formulation
Currently:
• Enabling techniques to improve bioavailability of BCS class 2 and 4 API’s.
• More recently formulation has become more focussed towards pharmacokinetics
• Requires a deeper understanding of the physicochemical properties of API’s in the presence
of the formulation ingredients for achieving required exposure levels
• Understanding the solubility and dissolution behaviour of API’s is an important part of
formulation design
6
Traditional dissolution testing is a quality performance test (drug release and QC)
Traditional Dissolution is not biorelevant…
Slide adapted from Gregory Amidon: “Oral Bioperformance and 21st century dissolution testing”?
7
New tools required for drug development
8
European IMI-OrBiTo* Program
* OrBiTo = Oral Biopharmaceutics Tools
http://www.orbitoproject.eu/
Platform for automated biorelevant dissolution and solubility testing with support for final dosage forms
http://www.sirius-analytical.com/products/inform
10
Sirius inForm - new instrument for use in Formulation studies
11
Automated dispensing of acid, base, solvents, buffers and SIFs during an assay
Stirrer, pH and UV probes. Sample vial in Peltier block. Automated sample addition.
Dispensers for Automated addition of media, titrants, solvents
12
Automated dispensing of acid, base, solvents, buffers and SIFs during an assay
Stirrer, pH and UV probes. Sample vial in Peltier block. Automated sample addition.
Robotic arms for vial handling and assay probe handling
13
Automated dispensing of acid, base, solvents, buffers and SIFs during an assay
Stirrer, pH and UV probes. Sample vial in Peltier block. Automated sample addition.
20 Vial autoloader, HPLC vial tray, sonicator, automated cleaning, vacuum manifold, tablet holder
14
The inForm measurement cell
15
pH electrode
UV probe
Temperature probe
Reagent capillaries
Stirrer
Inert gas
16
Sample tablet holder can be lowered from
above.
Basket for powders and
dosage forms.
Three ways to measure concentration
• pH-metric– Good for solutions whose pH is < 3 units from pKa
• In-situ UV– Good for drugs that absorb UV
• Automated Off-line sampling– Best method if samples are very turbid
17
18
Dru
g co
ncen
trati
on
Time
“Spring”
Equilibrium solubility
Diagram adapted fromBrouwers, J. et al. J. Pharm. Sci. 2009, 98 (8), 2549-2572.
Crystalline sample
“Spring” with “Parachute”
TRADITIONAL DISSOLUTION
“Spring and Parachute” model
19
Supersaturated, sample is precipitating
Subsaturated, sample is dissolving
Supersaturated, sample is fully dissolved
Precipitate appears
Equilibrium solubility
Time
0
2
4
6
8
10
0 30 60 90 120
Conc
entr
ation
(μM
)
Time (minutes)
20
Dissolution of Warfarin* powder at pH 3.1
* Warfarin is an acid, pKa 4.94
Equilibrium solubility
21
0 30 60 90 120Time (minutes)
0
100
200
300
400C
once
ntra
tion
(µM
)
Same graph, different scaling
Equilibrium solubility
0
100
200
300
400C
once
ntra
tion
of n
eutr
al s
pec
ies
(µM
)
22
Warfarin aqueous CheqSol: a “Spring”
Equilibrium solubility
Precipitate appears
Kinetic solubility
0 30 60 90 120Time (minutes)
0
100
200
300
400C
once
ntra
tion
of n
eutr
al s
pec
ies
(µM
)
23
0 30 60 90 120Time (minutes)
Warfarin and PVP*: a “Parachute”
* Polyvinylpyrrolidone. With thanks to Ashland Specialty Chemicals
24
How we measure concentrations by in-situ UV
In-situ UV
25
pH Versus Time
20:00 30:00Time(minutes:seconds)
3
5
7pH
N
O
O
CH3
OH
Cl
OCH3
IndomethacinAcid, pKa 4.1
IndomethacinTitration in linear buffer solutionMeasure spectra at each pH point
2 4 6 8pH (Concentration scale)
0.0
0.5
1.0
1.5
2.0
Abs
orba
nce
0
50
100
150
200
Species C
oncentration (µM)
5.11
250 300 350 400Wavelength (nm)
0.0
0.5
1.0
1.5
2.0A
bsor
banc
e
0
5000
10000
15000
20000
25000 Molar absorption (/cm
/M)
26
Molar Absorption Coefficients (MECs)
N
O
O
CH3
OH
Cl
OCH3
Comparing MEC with dissolution data
27
250 300 350 400Wavelength (nm)
0.0
0.5
1.0
1.5
Abs
orba
nce
N
O
O
CH3
OH
Cl
OCH3
28
inForm case studies
Client Evaluation 1 – A major US pharma company
29
• API is a weak acid with pKa = 9 and log P = 3
• Four forms supplied
– Crystalline API, powder
– Formulated crystalline API, extracted from a capsule
– Amorphous solid dispersion, powder
– Formulated amorphous solid dispersion, part of tablet
• Equal weight of API used in each experiment
• All experiments at 37°C
30
Dilution, FaSSIF introduced
Gastric period: Acetate-phosphate
buffer + HCl
Gastric emptying. pH raised with NaOH
Biorelevant dissolution. FaSSIF added
Intestinal period
31
Biorelevant dissolution
1
2
3
4
5
6
7
0
5
10
15
0 2 4 6
pH
Mas
s of d
isso
lved
API
(mg)
Time (hours)
Formulated crystal API
Crystal API
Solid dispersion
Formulated solid dispersion
pH
Crystalline samples
1
2
3
4
5
6
7
0
200
400
600
800
0 1 2
pH
Mas
s of d
isso
lved
API
(μg)
Time (hours)
Formulated crystal API
Crystal API
pH
Crystalline
Amorphous
Biphasic dissolution. Nonanol added
32
Gastric period: Acetate-phosphate
buffer + HCl
Dilution, lipid layer introduced
Gastric emptying. pH raised with NaOH
Drug dissolving in aqueous solution
Drug partitioned into lipid
Total amount dissolved
Intestinal period
1
2
3
4
5
6
7
0
5
10
15
0 1 2
pH
Mas
s of d
isso
lved
API
(mg)
Time (hours)
Crystal API, lipidFormulated crystal API, lipidFormulated crystal API, aq.Crystal API, aqueouspH
1
2
3
4
5
6
7
0
5
10
15
0 1 2
pH
Mas
s dis
solv
ed o
f API
(mg)
Time (hours)
Formulated solid dispersion, lipid
Solid disperion, lipid
Formulated solid dispersion, aq.
Solid dispersion, aqueous
pH
Biphasic dissolution
33
Crystalline API- may be overstimated
Formulated solid dispersion - may be overestimated
Crystalline
Amorphous
Client Evaluation 2 – Small US Pharma
• Controlled supersaturation• Solvent quench method: concentrations by UV• Two examples
– Bifonazole, Droperidol• FaSSIF and various additives used to test the
effect on supersaturation/precipitation rate
34
35
36
Tyndall-Rayleigh scattering correction
BEFORE
AFTER
OK for data analysis
37
Aqueous
Pluronics
PVP
Soluplus
FaSSIFHPMC
DroperidolBase, pKa 8.2Controlled supersaturation at pH 6.8
BifonazoleBase, pKa 6.6Controlled supersaturation at pH 5
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Summary of inForm PlatformThe Sirius inForm instrument is a useful tool for early development and
formulation scientists providing more predictive tools for drug performance
• Sirius inForm can set up a wide range of experimental conditions
• Automated biorelevant solubility & dissolution
• Automated biphasic dissolution
• Measuring supersaturation & precipitation behavior
• Dealing with turbidity
• Innovative tests for investigating IVIVC
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What about Protein formulations?
Introducing Sirius “Scissor”: Sub Cutaneous Injection Site SimulatOR
A New In Vitro Test for Injection Site Events for Biopharmaceuticals
Importance of Subcutaneous Injections
1989 to 2012 Biotechnology products (mostly proteins and peptides)
grew in number from 13 to 210 Sales increased to US$163 billion.
2001 to 2012 Biotech products accounted for 71% revenues for the
ten top-selling pharmaceuticals in 2012, up from 7% in 2001.
Move to subcutaneous (SC) injections Currently, ~ 40 protein and peptide drugs are given SC Therapies shifting care to home treatment will increase
this number of SC drugs
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Potential Issues for SC and IO injections
Current formulations are designed to• Keep API stable in a vial for several years• Minimize injection volume (high concentration)• Minimize pain upon injection
APIs are stressed upon injection by• Transition from formulation to homeostatic conditions immediately
after injection- Physical stress due to change in pH- Transition through isoelectric point?- Change in concentration of stabilizing agents- Altered interactions with stabilizing agents
42
Possible Events After Injection
There are currently no in vitro methods available to examine potential events that might be experienced by an API during its transition from an injected formulation to the steady-state conditions of the injection site environment.
43
What We Know and What is Needed
What we know• Site to site and patient to patient variability is seen for bioavailability (%BA) of
many biopharmaceuticals.• No animal model correlates to (%BA) observed in man.• Conditions/characteristics of physical and chemical environments of the
injection site are species specific.• Insolubility/precipitation upon injection can lead to cellular responses and
macrophage clearance.
What we need• A versatile in vitro model to examine the potential impact of specific, individual
post-injection events.• A dynamic system that emulates approximate time and conditions for post-
injection transitions.
44
Design of the Scissor instrument
45
Schematic of the Scissor system chamber featuring: injection cartridge acting as a simulated injection site; pH probe for monitoring the pH within the cartridge; light source and detector for monitoring aggregation events; chamber of physiological buffer; thermocouple and heater/stirrer
Sirius Scissor - Schematic
46
In vitro study of subcutaneous injection of two insulin formulations
Two insulin formulations; Insuman Rapid (fast acting) and Insuman Basal (slow acting)
Light %T of Insuman Rapid and Basal
PK of Insuman Rapid and Basal
Fraction appearing in ISF buffer (data by HPLC)
47
Pharma Study - Outcome Potentially Important for mAbs@
6hr
s
48
Our Approach
• No pre-clinical model has been identified that will correlate with human in vivo outcomes
• This model is not intended to examine cell-mediated or immune responses
• Our in vitro model simulates dynamic events at the site of injection site of a drug
• The model monitors ECM interactions, pH changes, protein turbidity, excipients fate and spectroscopic properties.
• The system models events for several hours at physiological conditions
• Future studies will focus on correlations pre-clinical human in vivo outcomes to establish a predictive tool.
49
Scissor - Acknowledgements
GenentechVikas SharmaStefan FicsherSreedhara AlavattamTom PatapoffAnn Daugherty
Sirius AnalyticalJohn ComerKarl BoxGeorge ButcherBrett Hughes
University of BathRandy MrsnyHanne KinnunenJenni SobleAlison EvansMatt Young
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•Thanks for listening!
•Any questions?
www.sirius-analytical.com
Make formulation decisions earlier with detailed bio-relevant data