XXIII NMR Symposium 14 January 2020, CERN

Drug Delivery Systems examined by NMR

Martina Vermathen, Ilche Gjuroski, Max Meier, Sara Pfister,
Luca Sauser, Julien Furrer

University of Bern, Switzerland, Department of Chemistry & Biochemistry

 Drug delivery systems

Porphyrinic Photosensitizers N
HN
NH
 Exhibit stacking in water N N
HN
NH
R N
 Aggregation reduces PDT efficiency
R

Polymeric Delivery Systems Chemical

Physical entrapment
bonding
 Monomerize porphyrin aggregates
 Enhance solubility in water
 Prolonged stabilty +

 Nanoparticles  EPR effect

1 – 1000 nm
 Improved photophysical properties
 Types of Drug delivery systems

 Intravenous application
 Dermal application

Single molecules
Micelles (BCMs) Liposomes Bicelles
Molecular networks

PVP Triblock-copolymers ABA Phospholipids

Cyclodextrines A B A

Building blocks PEG - PPG - PEG

 1
H NMR spectra H-5

Nanoparticles Chlorin e6 derivatives H-20

NH N
H-10
N HN
Colloidal dispersions Aggregation
CO2H
O
HO2C OH
1 – 100 nm
CETM
CEDM
CE4
CEMED
ArgCE
LysCE H2O
TyrCE

N
SerCE HN
H-10 H-20 H-5 NH
CE6 N
R

H-10 H-5 H-20

DMSO

5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm 10 9 8 7 6 [ppm]
 Monitoring drug encapsulation
CE4 Kolliphor 188 (KP)
1
H NMR spectra 1
H DOSY H-3 H-5
CH3

O O
H O OH
NH N x y x
CE4: R = CH3 H-20 H-10

D
log [m2/s]

x 64

PBS 5
-11.0
11 10 9 8 7 6 5 4 3 2 1 ppm
10 9 8 7 ppm
NH N
SerCE: R = CH2CONH-Ser 20 10
N HN CE4 + KP
-10.5

R CO2H KP
HO2C -10.0

11 10 9 8 7 6 5 4 3 2 1 ppm
PBS CE4 / DMSO -9.5
10 9 8 7 ppm
10 8 6 4 2 ppm
I. Gjuroski et al., J. Controlled Release (2019), 316, 150-167
 Monitoring drug encapsulation
1
H 1H NOESY
PPG-CH3
CE4 CH3 KP
3 5
O O
H O OH
+ CE4 NH N x y x
20 10
+ SerCE
H-5 H-10
H-20 H-3’

F1 ppm
1.6 1.4 1.2 1.0 0.8 ppm
-CH3

1
KP intermolecular

2
PPG-CH3
PEG-CH2
CH3
intramolecular

3
O O
H O OH

4
x y x

PEG PPG PEG H-2’ CE4 – KP 3 : 10

10 9.5 9.0 8.5 ppm
I. Gjuroski et al., J. Controlled Release (2019), 316, 150-167
 Monitoring drug encapsulation
N HN

CH3

O O
H O OH R CO2R
x y x

RO2C

Tuning the micelle properties logP

HLB SerCE CE4 CEMED CEDM CETM

PEG PPG PEG

P84
L64
F127
F108
KP188

 HLB PBS
 MW 9.0 8.0 9.0 8.0 9.0 8.0 9.0 8.0 9.0 8.0 [ppm]
 PPG units
S. Pfister, L. Sauser, I. Gjuroski , J. Furrer, M. Vermathen, J. Porphyrins Phthalocyanines (2019) in press
 Monitoring drug encapsulation
N HN

CH3

O O
H O OH R CO2R
x y x

RO2C

Tuning the micelle properties logP

logP
HLB SerCE CE4 CEMED CEDM CETM
PEG PPG PEG P84
L64 P84 KP188 F127 F108
35
P8470
30 60 L64
25 L6450

PPG units
F127
HLB

20 40
F127
15 30
F108 F108
10 20
5 10
KP188 KP188
0 0
0 2 4 6 8 10 12 14 PBS
16
MW [kDa] 0 20 40 60 80 100
9.0 8.0 9.0 8.0 9.0 8.0 9.0 8.0 9.0 8.0 [ppm]
xCE encapsulated [%]

S. Pfister, L. Sauser, I. Gjuroski , J. Furrer, M. Vermathen, J. Porphyrins Phthalocyanines (2019) in press

 Comparing micelles and PVP
Ce6 5 Micelles (KP) PVP

NH N
20 10
Ce6-Aminoacid I [au]
PBS H-20 H-20
H-5 H-10 H-20 conjugates t 6 H-10
KP H-10 H-5 5
* Ce6 8d 4
PVP
PBS
3
KP KP
2
Ser-Ce6 PVP
PVP * 0d 1
KP 0
Intensity decrease 0 2 4 6 8 t [d]
* Lys-Ce6
PVP
KP Micelles and PVP:
* Tyr-Ce6
PVP Þ Good encapsulation for Ce6-AAs
KP Þ Prevention of aggregate growth over time
* Arg-Ce6
PVP I. Gjuroski, J. Furrer, M. Vermathen, ChemPhysChem (2018), 19, 1089-1102
ppm M. Hädener, I. Gjuroski, J. Furrer, M. Vermathen, J. Phys. Chem. B (2015), 119, 12117-12128
10 9.5 9
Comparing micelles and PVP: Dynamics

D D
log [m2/s] log [m2/s]

PVP KP
-11.0 -11.0
SerCE + PVP CE4KP
+ KP
x 64
PVP -10.5 -10.5
SerCE + KP
-10.0 -10.0

-9.5 -9.5
SerCE SerCE
Strong binding to polymer -9.0 Dynamic
Stronger exchange
binding for hydrophobic -9.0
CE4
10 8 6 4 2 ppm 10 8 6 4 2 ppm

I. Gjuroski, J. Furrer, M. Vermathen, ChemPhysChem (2018), 19, 1089-1102

M. Hädener, I. Gjuroski, J. Furrer, M. Vermathen, J. Phys. Chem. B (2015), 119, 12117-12128
 Comparing micelles and PVP: Release

Release profiles (PBS, 37°C) PVP SerCE CE 4

SerCE-KP + Tf
Chlorin release [%]

CE4-KP/PVP
10
SerCE-PVP + HSA

1 PBS
5 10 15 20 25 30
Time [h] 10 9.6 9.2 ppm 10 9.6 9.2 ppm

0
KP SerCE CE 4
 PVP: + Tf
Reduced reactivity towards proteins
+ HSA
 Micelles (KP):
Release in favor of HSA binding
PBS
10 9.6 9.2 ppm 10 9.6 9.2 ppm

I. Gjuroski et al., J. Controlled Release (2019), 316, 150-167

Bicelles for topical drug delivery

NH N
Topical PDT 5-ALA PPIX ?
N HN

Stratum Corneum (SC)

Epidermis
Intercellular route
Dermis
through lipid matrix

Hypodermis

Flexible liposomes Bicelles

 Bicelles for topical drug delivery
Phospholipids: long- and short-chain
 Small size
R
 Disc-shaped R

 Biocompatible
Bilayer DMPC (C-14) R = -(CH2)12CH3
Bilayer-forming
Bicelles
Micelles DHPC (C-6) R = -(CH2)4CH3
Edge-forming

DHPC DMPC
DHPC-DMPC
50 mM, q = 0.5

5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 ppm
 Bicelles for topical drug delivery
Bicelles 150 mM DHPC/DMPC q = 0.5 H1H NOESY:
1 5

with 5 mM xCE CE4 – DMPC/DHPC 82

N HN 20 10

H-5
H-10 181 H-20

CO2H H

(-CH2CO2H) ω-CH3
HO2C
Ce6 TME Intermolecular NOEs
-(COMe)3 CE4 DMPC
-(CH2)n

Ce6 DME
-(COMe)2

H-82
CE4
H-181
CE4 Intramolecular NOEs
10.0
10 9.5 9.0
9 8.5 8.0
8 7.5 7.0
7 6.5 6.0
6 ppm
ppm
 Bicelles for topical drug delivery: Outlook

Modifications:
Long chain phospholipids
PEG-ylated PLs
Charged PLs
PL- transition temperature
Additives

Short chain phospholipids q-value

 Prevent fusion
Low water solubility
Low cmc  Keep bicelle morphology
 Enhance stability
 Enhance encapsulation
Acknowledgement

Financial Funding:
Swiss National
Science Foundation THANK YOU
SNF
FOR
YOUR ATTENTION