Small Ions, Large Antibodies, and Everything in between

Small Ions, Large Antibodies, and
Everything in between
Real-Time, Target-Site Monitoring with Open Flow
Microperfusion
PK/PD at target-site
2
!
Isn’t blood just perfect?
Microdialysis
working principle
3
Interstitial fluid
c=0
ISF is filtered by a membrane
Interstitial fluid
c≠0
Microdialysis (MD)
overview
Substance / Drugs …
MD
… small
YES
… hydrophilic(small)
YES
… larger + large
YES
& NO
… lipophilic
(super lipophilic)
NO
… protein-bound
NO
… (nano)carrier / cells
NO
membrane,
nm-µm pores
100 µm
5
Let’s get rid of the membrane…
Over 10 years ago
2014
Speakers
6
Dr. Frank Sinner
Deputy Director HEALTH
JOANNEUM RESEARcH
Austria
Europe
Dr. Thomas Alsted
Research Scientist
Novo Nordisk A/S
Denmark
Europe
Dr. Gerard Bruin
Senior Investigator II
Novartis Pharna AG
Switzerland
Europe
Open Flow Microperfusion (OFM)
Open Flow Microperfusion for Target-Site PK/PD
Monitoring of a Wide Range of Substances
Open Flow Microperfusion
working principle
8
Interstitial fluid
c=0
ISF is diluted but unfiltered
Interstitial fluid
c≠0
Open Flow Microperfusion
overview
Substance / Drugs …
MD
OFM
… small
YES
YES
… hydrophilic(small)
YES
YES
… larger + large
YES
& NO
YES
… lipophilic
(super lipophilic)
NO
YES
(?)
… protein-bound
NO
YES
… (nano)carrier / cells
NO
YES
100 µm openings
in exchange area
of OFM probe
Open Flow Microperfusion
lipophilic substances
Successfully sampled drugs
using OFM
Substance Doxorubicin BCT 194
Fluorescein Betamethason-17-valerate Clobetasol-17-propionate Fluoxetine Amitryptiline Lipophilicity
Log (P) 1.3
3.1
3.4
3.6
4.0
4.6
4.9
Open Flow Microperfusion
high-molecular weight substances
Successfully sampled analytes
using OFM
Substance BDNF IL-1b TNF - alpha Albumin Humanized antibody
Nanoparticles Entire cells
Size
14 kDa 31 kDa 51 kDa 66 kDa ~150 kDa
~100 nm ~ 8 µm
Open Flow Microperfusion
12
set-up
Clinical
•  Skin tissue
•  Fat tissue
Pre-clinical
•  Skin tissue
•  Fat tissue
•  Brain tissue
aOFM / dOFM models
ex vivo compound PK testing
13
!
Available models:
pig and human skin
!
Duration:
1-2 days
!
Application sites:
from 2 up to 8 sites per skin flap
!
OFM material:
same as in preclinical and clinical models
!
Time resolution:
determined by analytics (5 to 120 min)
aOFM / dOFM models
preclinical in vivo testing
14
!
Available models:
pigs, rats and mice
!
Duration:
up to 14 hours
!
Application sites:
up to 6 probes per rat; up to 24 probes per pig
!
OFM material:
same material for preclinical and clinical
!
Time resolution:
determined by analytics (5 to 120 min)
cOFM models
preclinical in vivo testing
15
!
Available models:
rats and mice
!
Duration:
up to 10 hours
!
Application sites:
up to 2 probes per rat
!
OFM material:
preclinical only
!
Time resolution:
determined by analytics (5 to 120 min)
aOFM / dOFM models
clinical in vivo testing
16
!
Available models:
healthy volunteers and
patients
!
Duration:
up to 48 hours
!
Application:
up to 15 probes
per subject
!
OFM material:
same material for preclinical and clinical
!
Time resolution:
determined by analytics
(5 to 120 min)
Open Flow Microperfusion
strengths
17
1.  Minimal tissue response at target site
2.  PK/PD of lipophilic APIs at target site
3.  Bioequivalence at target site
4.  PK of albumin bound APIs at target site
5.  PK/PD of large APIs at target site
6.  OUTLOOK: Metabolomics at target site
1. Minimal tissue response at target site
cOFM
18
Tissue response to a microdialysis and cOFM probe 2 weeks after probe implantation
µD
probe
trauma layer surrounding microdialysis membrane
(Benveniste and Diemer 1987)
cOFM
probe
no trauma layer surrounding cOFM
No continuous glial scar formation around the cOFM probe
after 30 days of implantation.
18
2. PK/PD of lipophilic API’s at target site
19
dOFM
2. PK/PD of lipophilic APIs at target site
highly potent corticoid
20
!
Clinical study in psoriatic patients
!
12 dOFM probes per subject
!
26 h dOFM sampling
on day 1 and day 14
!
Analysis
!
!
Clobetasol-17 propionate (PK)
10 cytokines (PD)
lesion1
drug
lesion2
vehicle
unaffected1
drug
unaffected2
vehicle
2. PK/PD of lipophilic APIs at target site
exemplary CP-17 profiles
Concentration ng/ml
PK
2
ofm04
lesional
ofm05
1.5
Concentration [ng/ml]
1st subject - 1st day - 1st dose (dOFM: 26hrs)
!
ofm06
1
0.5
non lesional
ofm11
1.5
ofm12
1
1.17mm
2
600
ofm04
ofm05
500
ofm06
3
4
5
6
7
0
8
700
sample interval
Concentration pg/ml
1
700
concentration [pg/ml]
ofm10
1.07mm
0.5
0
PD
2
400
300
200
100
0
1
2
3
ofm10
600
ofm11
500
ofm12
4
5
6
7
8
6
7
8
sample interval
400
300
200
100
0
1
2
3
4
5
sample interval
6
7
8
1
2
3
4
5
sample interval
2. PK/PD of lipophilic APIs at target site
summary
22
!
Summary and Conclusions
!
!
!
!
26 h x 12 probes with
wearable set-up
high reproducibility between
probes at same site
PK (CP17)
!
detectable in dermis
after ~12 h
!
differences found between
L-D, L-V, NL-D, NL-V
PD
!
Four cytokine profiles are detectable and altered by drug
3. Bioequivalence at target site
dOFM
23
!
Topical generics can only show bioequivalence in
clinical endpoint studies
!
Need to facilitate the development of generics
!
Ongoing FDA co-financed project to investigate
bioequivalence and non-bioequivalence in skin of
topical drugs using dOFM
4. PK of albumin bound APIs at target site
aOFM
24
!
Preclinical study to investigate
!
PK at target site à profile of different insulins in
adipose tissue after i.v. administration
!
PK at target site versus PD in blood
Preclinical study presented by Thomas Junker Alsted, Novo
Nordisk:
“Using OFM in adipose tissue to correlate pharmacokinetics of
insulin analogues to pharmacodynamics”
5. PK/PD of large APIs at target site
dOFM
25
!
Clinical study to investigate
!
PK at target site à the absolute concentration of
secukinumab in skin after s.c. administration
!
PD at target site à effect of secukinumab on IL17 and
downstream biomarker
Clinical Study presented by Gerard Bruin, Novartis:
“Absolute quantification of the anti-Il-17A antibody secukinumab
and relevant biomarkers in skin by dOFM; an assessment in
healthy volunteers and patients with moderate-to-severe plaque
psoriasis”
6. OUTLOOK: Metabolomics at target site
OFM meets metabolomics
!
Combination of OFM and metabolomics
!
to investigate the pathogenesis of diseases and the
influence of APIs on the metabolome
Thank you for your attention
JOANNEUM RESEARCH
Forschungsgesellschaft mbH
HEALTH
Institute for Biomedicine
and Health Sciences
Dr. Frank Sinner
Deputy Director
[email protected]
Neue Stiftingtalsstrasse 2
8010 Graz
+43 316 876-4000
www.joanneum.at/health
4. PK of albumin bound API’s at target site
aOFM
28
!
Pre-clinical Study to investigate
!
PK at target site à profile of different insulin’s in
adipose tissue after i.v. administration
!
PK at target site versus PD in blood
Pre-clinical Study presented by Thomas Alsted, Novo Nordisk:
“Using OFM in adipose tissue to investigate pharmacokinetics of
an albumin associated insulin analogue”
5. PK/PD of large APIs at target site
dOFM
29
!
Clinical study to investigate
!
PK at target site à the absolute concentration of
secukinumab in skin after s.c. administration
!
PD at target site à effect of secukinumab on IL17 and
downstream biomarker
Clinical Study presented by Gerard Bruin, Novartis:
“Absolute quantification of the anti-Il-17A antibody secukinumab
and relevant biomarkers in skin by dOFM; an assessment in
healthy volunteers and patients with moderate-to-severe plaque
psoriasis”
Open Flow Microperfusion
Conclusion
30
!
OFM shows..
minimal tissue reactions
! excellent acceptance by subjects
!
!
OFM allows sampling of diluted interstitial fluid for..
PK/PD of lipophilic APIs at target sites
! PK/PD of large APIs at target sites
!
31
Tissue response to
cOFM probe implantation
Quantification of
astrocytes
15 days after cOFM
probe implantation
Quantification of
microglia
15 days after cOFM
probe implantation

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