hERG, temperature & pharmacology

Ion Channel Retreat Vancouver August 9 - 11 2017

Anders Lindquist

Distributor Sophion Direct Sales Office

Sales Office/demo lab Boston, MA USA

Sophion Headquarters, R&D, TS &

Manufacturing Copenhagen, Denmark

Sales Office Shanghai, China

Sales Office/demo lab Tokyo, Japan

Location Total Head Count

Copenhagen, Denmark 44

Tokyo, Japan 4

Shanghai, China 1

Boston, USA 6

Cambridge, UK 1

Total 56

Global Company with Local Reach

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Why hERG

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Kannankeril et al. Pharmacolog Rev 2010

hERG pharmacology?

4 Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Hardly the exciting NKOTB anymore • Center stage for safety screening

and for the CiPA initiative. • hERG pharmacology is a complex

function of voltage protocol, temeprature, solutions and other factors.

.

IC50 cisapride as f(VP, T, ...) Milnes et al. JPTM, 2010

QUBE Automated patch-clamp system

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• 384 well instrument w/ 384 amplifiers

• Voltage-gated, ligand-gated and current clamp recordings

• Combine freely in the same experiment

• Temperature control • Both heat and cool

• Unattended operation and up to 18.000 wells tested/day

• Single or multi-hole micro-fluidic consumables

QChip 384

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Individual electrode pairs for each well

Ensure stable recordings over a long period of time

Instantly ready for experiments

Maintenance free

Liquid interface from the top

Electronic interface from the bottom

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Qube Temperature Control

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Control Unit

Temperature Unit Biochip Interface

Pump

Sensor Heating/ cooling system

hERG tail current vs temperature

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Plate-view of standard protocol currents

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20C

34C 31C

25C

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Steady-state activation

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Temperature V1/2 (mV)

20 8.7

25 1.3

31 -4.6

34 -11

20C

34C

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Activation cont’d

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Temperature tau (ms)

20 820

25 320

31 100

34 50

20C

34C

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Steady-state inactivation

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20C

34C

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Temperature V½ (mV)

20 -52

25 -44

31 -37

34 -28

Pharmacology

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Quinidine 10 µM @ 25 C

Astemizole Bepridil

Quinidine Verapamil

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

Astemizole 41 nM @ 25 C

Temp. (C) IC50 (µM)

20 0.008

25 0.008

Temp. (C) IC50 (µM)

20 0.25

25 0.18

Temp. (C) IC50 (µM)

20 1.2

25 1.5

Temp. (C) IC50 (µM)

20 0.51

25 0.44

Verapamil Quinidine

Pharmacology

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hERG and temperature

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• No simple yes and no answer but depends on the circumstances there are temperature dependent compounds but a minority? longer protocols tend to decrease temperature-dependent potency difference.

Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017

• Possible to run temperature controlled testing on routine basis • Find other primerary drivers for your project than ”just” potency

Milnes et al. JTPM, 2010

Click to edit Master subtitle style

Thank you!

Questions?

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Acknowledgements Mette Christensen Pia Andersen Daniel Sauter Mads Korsgaard Rasmus Jacobsen Melanie Schupp Weifeng Yu

Structure by Wang et al.

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Cryo-EM Structure of the Open Human Ether-a-go-go-Related K+ Channel hERG Wang W, MacKinnon R Cell (2017) 169 p.422-430

Inactivation and recovery from inactivation

17 Anders Lindqvist Ion Channel Retreat Vancouver August 9 - 11 2017