The role of helper lipids in the design of lipid ...

The role of helper lipids in the design of lipid nanoparticle technology for nucleic acid delivery

PHOSPHOLIPID

SYMPOSIUM

September 14th, 2021

Dominik Witzigmann, PhDNanoVation Therapeutics Inc.

NanoMedicines Innovation Network (NMIN)

CRS Focus Group Gene Delivery and Gene Editing

International Symposium on Phospholipids 2019

2

Phospholipid Research Center -Researcher’s Day 2021

3

4

COVID-19 changed the world and…

Kulkarni & Witzigmann et al. 2021 Nature Nanotechnology

…the landscape of LNP-nucleic acid therapeutics

Silencing & Gene Expression

5

➢ The Importance of Lipid Nanoparticle Design• General Composition of approved LNP-RNA systems

➢ The role of helper (phospho)lipid• RNA entrapment, stability, tropism, potency

I

II

➢ Future Perspectives• The Liver, Vaccines and Beyond

III

A Breakthrough Technology

LNP Technology Enabling Gene Therapy

6

Approved LNP-nucleic acid therapeutics


Comirnaty

(BNT162b2)

ALC-0315

DSPC

Cholesterol

ALC-0159

mRNA

Onpattro

(patisiran)

MC3

DSPC

Cholesterol

PEG-DMG

siRNA

mRNA-1273

SM-102

DSPC

Cholesterol

PEG-DMG

mRNA

7Schoenmaker, Witzigmann et al. 2021 Int. J Pharm.

Witzigmann & Kulkarni et al. 2020 Advanced Drug Delivery Reviews



Comirnaty

(BNT162b2)

ALC-0315

DSPC

Cholesterol

ALC-0159

mRNA

Onpattro

(patisiran)

MC3

DSPC

Cholesterol

PEG-DMG

siRNA

mRNA-1273

SM-102

DSPC

Cholesterol

PEG-DMG

mRNA





Comirnaty

(BNT162b2)

ALC-0315

Onpattro

(patisiran)

MC3

mRNA-1273

SM-102



Branched lipid tails improve mRNA delivery

pKa 6.75 pKa 6.09pKa 6.44



Comirnaty

(BNT162b2)

ALC-0315

DSPC

Cholesterol

ALC-0159

mRNA

Onpattro

(patisiran)

MC3

DSPC

Cholesterol

PEG-DMG

siRNA

mRNA-1273

SM-102

DSPC

Cholesterol

PEG-DMG

mRNA





I

II


III



11

Impact on RNA entrapment


Helper lipids are vital for

entrapment and

physicochemical properties

12

Kulkarni & Witzigmann et al.

2019 Nanoscale

Impact on stability


Phospholipid/cholesterol

free systems are not

stable in serum

13

Unpublished Data

1 h 24 h

Sato et al. Acta Biomater. 2020

doi: 10.1016/j.actbio.2019.11.022

Unpublished Data

Impact on cellular tropism


Incorporation of anionic phospholipids into

LNP systems changes cellular tropism14

Unpublished data which cannot

be shared at this point. Please

contact me directly if you have

any questions. Happy to jump on

a call or answer via email.

Impact on tissue tropism


Cationic LNP systems preferentially transfect the lung,

e.g. alveolar macrophages, but result in enhanced toxicity15

Cheng et al. 2020

cDC1s

cDC2s

pDCs

Mac

sM

os

cMos

pMos

Neu

ts

B cells

T cells

HSPCs

0

20

40

60

80

Bone Marrow

% T

hy1.1

+

****

**** ****

****

****

**

****

*****

****

cDC1s

cDC2s

/Mac

s

pDCs

Kup

ffer C

ells

Neu

ts

B cells

T cells

CD45

-

0

50

100

150

Liver

% T

hy1.1

+****

********

******** *

** *

cDC1s

cDC2s

/Mac

s

pDCs

Alv M

acs

Neu

ts

B cells

T cells

CD45

-

0

10

20

30

40

50

Lungs

% T

hy1.1

+

*

****

****

**** ****

****

**

CTRL

BioNTech

BioNTech

cDC1s

cDC2s

pDCs

Mac

sM

os

cMos

pMos

Neu

ts

B cells

T cells

HSPCs

0

20

40

60

80

Bone Marrow

% T

hy1.1

+

****

**** ****

****

****

**

****

*****

****

cDC1s

cDC2s

/Mac

s

pDCs

Kup

ffer C

ells

Neu

ts

B cells

T cells

CD45

-

0

50

100

150

Liver

% T

hy1.1

+

****

********

******** *

** *

cDC1s

cDC2s

/Mac

s

pDCs

Alv M

acs

Neu

ts

B cells

T cells

CD45

-

0

10

20

30

40

50

Lungs

% T

hy1.1

+

*

****

****

**** ****

****

**

CTRL

BioNTech

BioNTech

Unpublished data which cannot

be shared at this point. Please

contact me directly if you have

any questions. Happy to jump on

a call or answer via email.

Impact on administration route


Different LNP systems

are needed dependent

on target tissue and

route of administration

16

Unpublished Data

Nasal Cavity Lung



I

II


III



17

LNPs: The Next-Generation Pharmaceuticals


18

Lipid nanoparticles (LNPs) have been developed for siRNA delivery to the

liver and currently similar formulations are used for mRNA vaccines

LNPs: The Next-Generation Pharmaceuticals


19

Lipid nanoparticles (LNPs) have been developed for siRNA delivery to the

liver and currently similar formulations are used for mRNA vaccines

Need for developing LNP formulations specifically designed and optimized for (extrahepatic)

tissues, payloads and applications

[email protected]

THANK YOU

https://www.nanomedicines.ca/

NanoMedicines Innovation Network Pieter R. Cullis and team

Christian Kastrup and teamMarco Ciufolini and team

Karen ChanValentina Francia

Jerry LeungSarah Hedtrich

Colin RossKenneth Harder

NanoVation TherapeuticsJayesh KulkarniAnthony TamDaniel Kurek

UPennNorbert Pardi

University of ZurichGerald Schwank and team

Leiden UniversityFred Campbell and team

Eindhoven UniversityRoy van der Meel

TAKE HOME MESSAGE

From the first-ever siRNA drug to COVID-19 mRNA vaccines

Lipid nanoparticles are revolutionizing medicines

STAY TUNED FOR THE FUTURE!

21

The role of helper lipids in the design of lipid ...

Documents