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Article:

Manoharan, Anand, Rayner, Peter J. orcid.org/0000-0002-6577-4117, Iali, Wissam orcid.org/0000-0002-9428-2023 et al. (3 more authors) (2017) Achieving Biocompatible SABRE : An in vitro Cytotoxicity Study. CHEMMEDCHEM. ISSN 1860-7179

https://doi.org/10.1002/cmdc.201700725

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FU

A

A

B

[a]

[b]

Ab

Si

th

Es

ap

SABR

SABR

m

we

de

bip

in

In

Cl

dise

an

an

al

in

re

di

Po

ne

tu

no

pa

dist

Th

Ex

FULL PAP

Achievin

Anand Manoh

B. Duckett*[a

] Dr. A. Manoh

Department o

University of

Heslington

York

YO10 5DD (U

E-mail: simon

] Prof. V. H. Pe

School of Bio

University of

Southampton

Supporting in

bstract: Produ

Signal Amplificati

the door to simple

Essential to

pplications is th

SABRE reaction

SABRE approach

main cause of ob

e develop tw

eactivation and

phasic catalysis

vivo study.

Introduction

linical Magnetic

isease diagnosi

nd field gradie

natomy. Howeve

ll clinical applica

n the body. Hyp

esponses into

iagnostic tech

Polarization (DN

ecessary to tra

umors[1] whilst

oble gases has

An alternative

arahydrogen (p

istribution witho

This technique i

Exchange (SAB

Figure 1: Sch

PER

ng bioco

oharan,[a] Petea]

haran, Dr. P. J. Ra

of Chemistry

York

(UK)

n.duckett@york.ac

Perry

ological Sciences

Southampton

n (UK)

nformation (SI) for

uction of a bioc

ion By Reversi

e clinical diagno

successful pr

he determinatio

mixture. Here

h using in vitro

bserved toxicity

wo catalyst r

d ion-exchange

s. These routes

n

c Resonance I

sis. It utilizes st

ients to give

ver, due to MR

cations detect w

perpolarization

strong signals

chniques. For

NP) has been u

ack the in vivo m

Spin Exchan

s allowed the di

ve low cost app

p-H2) to creat

out changing o

is known as S

SABRE) and is sho

Schematic repre

ompatib

eter J. Rayne

ayner, Dr. W. Iali,

ac.uk

r this article is give

compatible hype

ible Exchange (

osis via magneti

rogression to

on of the toxic

we exemplify th

o cell assay. We

is due to the SABR

removal metho

e chromatograp

s produce a bol

Imaging (MRI)

strong magnetic

detailed imag

RI’s inherent low

water due to its

n methods turn

s and thus ope

r example,

used to create

metabolism of

nge Optical Pu

iagnosis of pul

proach to hype

te a non-Boltzm

of the identity m

Signal Amplific

own schematica

esentation of the

ble SAB

ner, [a] Wissa

Dr. M. J, Burns, P

en via a link at the

erpolarized bol

(SABRE) could

tic resonance im

pre-clinical/c

cology profile o

the cytotoxicity

We conclude th

SABRE catalyst.

ods, one inv

phy and the se

lus suitable for

is at the forefro

c fields, radio w

ges of the hu

w sensitivity, a

s high concent

n typically weak

en the door to

Dynamic Nu

e the signal str

pyruvate in pro

Pumping (SEO

lmonary disease

erpolarization ut

zmann nuclear

molecule of inte

cation by Reve

cally in Figure

SABRE effect

1

BRE: An

am Iali, [a] M

Prof. S. B. Duckett

e end of the docum

lus by

d open

maging.

/clinical

of the

of the

that the

t. Thus

volving

econd

future

ont of

waves

human

almost

ration

k MRI

o new

uclear

rength

ostate

EOP) of

ses. [2]

utilizes

r spin

erest.

versible

1. [3] It

can e

Reso

such

laten

typica

Simu

transf

low m

the

subst

A

nicot

simu

detect

in thi

to 50

appro

for in

meth

pharm

studi

Th

route

prima

throu

as si

The

proce

hydro

d2-M

cytot

An

SABR

the a

inform

There

toxici

subst

SABR

huma

catal

that co

n in vitr

Michael J. Bur

t

ment.

enhance the si

sonance (NMR)

ch as 1H, 13C, 1

nt polarization

cally [Ir(H)2(Su

ultaneous bind

sfer of polariza

magnetic field

catalytic comp

strate in solution

A recent study

tinamide and

ultaneously imp

ction and reduce

is study was m

0% polarizatio

oaching 2 minu

n vivo detectio

hyl nicotinate

maceutics, the

ied.

The isotopic lab

e to modify

arily affects the

ugh a pathway

significant rate ch

metabolic pat

ceeds via forma

oxy species ar

MN in compariso

toxicity analysis.

Another potent

BRE technique

adverse effects

mation is kn

refore we prese

city of the met

strate effects.

BRE reaction by

an cell lines a

lyst from soluti

could be progre

tro cytoto

urns, [a] V. H

signals detected

) spectroscopy15N and others.

of p-H2 throug

Sub)3(IMes)]Cl

ding of the su

zation through t

ds.[6] Subseque

plex allows th

n.

showed the e

d methyl

prove their SAB

ce magnetic re

methyl-4,6-d2-nico

on in conjunct

nutes.[7] This ma

ion. However,

e (MN) in

e effect of sele

belling of drug

their safety a

e biological fate

that involves h

changes occur

thway of nico

ation of the N-o

re also known.

son to MN is

s.

tial source of

is the iridium

s of iridium sa

known about

sent a thorough

tal catalyst in

We determin

by performing i

and present a

ion in order to

essed to in vivo

toxicity s

Hugh Perry[

d by MRI and N

across a wide

s.[4] It works by

gh binding to a

l,[5] as hy

substrate allow

the scalar cou

ent substrate d

he build-up o

effect of 2H-lab

nicotinate m

ABRE-enhanced

elaxation. The o

cotinate (d2-MN

ction with T1 r

makes d2-MN an

despite some

cosmetic

ective deuterati

molecules is a

and/or efficac

e of drugs that

hydrogen-carbo

due to the kine

cotinic acid der

oxide, though ro

. For this reaso

determined he

toxicity that

catalyst. Whilst

salts has been

organo-iridium

h investigation

n conjunction w

ne the biocom

in vitro cytotox

a method for

o create a bioco

vo measuremen

study[b] and Simon

Nuclear Magne

e range of nucl

y harnessing t

a metal cataly

ydride ligand

ws spontaneo

upling network

dissociation fro

of hyperpolarize

belling a series

molecules ca

d NMR and M

optimal substra

N) which gave

relaxation valu

n ideal candida

e applications

and veterina

ion has not be

a well-establish

cy.[8] Deuterati

t are metabolize

on bond breaki

etic isotope effe

rivatives typica

outes involving

son the toxicity

ere using in vi

arises from t

st some data

n reported[9], lit

m complexes[

into the potent

with solvent a

mpatibility of t

xicity analysis

deplet ion of t

compatible bol

nt.

on

etic

clei

the

yst,

ds.

ous

at

om

zed

s of

can

MRI

ate

up

ues

ate

of

ary

een

shed

ion

zed

king

ect.

cally

g 6-

y of

vitro

the

on

ittle[10].

ntial

and

the

on

the

lus

FU

R

Ev

M

hig

bi

co

to

ef

do

bi

fo

Ho

un

asc

to

in

est

eit

th

ob

wa

an

of

sim

IC

use

th

di

st

wh

pr

an

m

acr

Ef

SA

alco

so

aq

re

Ta

A5

MC

He

MD

FULL PAP

Results and

Evaluating the

Methyl-4,6-d2-nico

ighest 1H pol

iological persp

cosmetics due t

opical applicati

ffects at lower

oses.[11] Given

iological applic

or probing on

owever, wheth

nclear. In this st

scertain the bro

In order to

oxicity we beg

nhibitory conce

stablished cance

ither MN or d2-

he viability was

btained for each

as found to ran

nd interestingly

f the IC50 value

milar to MN. Im

C50 values (Ta

sed in a typica

hese results a

irectly in, the h

ill unclear how

here different

rovide a refere

nalysis. From t

methyl nicotinate

cross the cell li

Effect of SABR

SABRE induced

lcohol solvents

some applicatio

queous solutio

educed.[14] In o

able 1. Comparing

549

CF7

eLa

DA-MB231

PER

d Discussio

cytotoxicity of

cotinate (d2-M

larization leve

pective, MN is

to its percutan

ion and more

r doses and i

the substantia

cability, d2-MN

in vitro/in v

her d2-MN retai

study, we utilize

oader SABRE t

o establish the

gan by deter

centration) of

cer cell lines o

-MN at varying

s assessed by

ch cell lines is s

nge between 1

y, despite the d

es indicated th

mportantly, bot

able 1) and ar

al SABRE rea

are based on

high volume of

w the effect wou

solvents are

ence IC50 value

his analysis, w

e has no quant

ines studied he

RE solvents on

d polarization

s such as metha

ons can produ

on, though en

order to create

ring the IC50 of MN v

Methyl Nicot

(MN)

13.6

33.3

21.9

12.6

ion

of SABRE subs

MN) has been r

vels by SABRE

s widely used

neous penetrat

eover known f

inflammatory r

al SABRE enha

is considered

vivo tests for

ins the charact

ze d2-MN as a

toxicology prof

effect of select

rmining the I

MN and d2-M

of human origi

g concentration

y the MTT met

shown in Table

12.6-33.3 mM a

deuterium labe

hat the toxicity

th substrates h

re within the co

action (mM). It

the solubility

, the cell cultu

uld be under co

used. Nevert

e for optimizing

we conclude tha

tifiable effect in

ere.

on cell viability

n levels are t

anol-d4 or etha

uce hyperpola

nhancement l

e a biocompat

vs. d2-MN in huma

IC50 (mM)

tinate Meth

bstrate

reported to give

E to date. [7]

d as rubefacie

ion properties

for its vasodi

response at h

ancement level

d an ideal cand

SABRE dete

cteristics of MN

a model substra

ile.

ctive deuteratio

IC50 (half ma

MN. For this,

in were treated

ns for up to 48

thod. [12] The re

e 1. The IC50 fo

across the cell

elling, a compa

levels of d2-M

have millimolar

concentration r

is noted here

y of the subst

ure medium an

conditions of SAB

theless, these

g SABRE for fu

at deuteration

n modulating to

typically highe

anol-d6.[7, 13] How

arized substrat

levels are typ

tible hyperpola

an cancer cell line

hyl-4,6-d2-Nicotina

(d2-MN)

17.4

25.2

14.9

12.3

2

ve the

From

ent in

upon

ilatory

higher

ls and

didate

ection.

is still

rate to

on on

aximal

s, well

d with

h and

results

or MN

l lines

arison

MN are

(mM)

range

e that

strates

nd it is

SABRE,

data

urther

of the

oxicity

est in

wever

tes in

pically

arized

bolus

hype

dilutio

To

perfo

cells

D2O.

signif

cultu

over

show

durat

(Figu

ethan

over

solve

the re

evide

highe

solve

cytot

achie

durat

perfo

As

deter

quan

polar

mM

(30:7

found

at 9.4

supp

reduct

d6 as

enha

furthe

press

ongo

es.

ate

Fig

cell

h o

solv

and

**P<

viable);

s with high pola

erpolarization st

ion with D2O.

To assess th

ormed an appro

which were tr

. As shown in

ificantly reduce

ure medium an

the same tim

wed no chang

tions to 24 h,

ure 2 B). Where

nol-d6 in D2O (

long treatmen

vents mixtures (

results indicated

ent when the

er than or equa

vent mixtures is

toxicity analysi

eve an optimal

tion of exposu

orming SABRE

As the optim

rmined here i

ntify the effects

rization of d2-M

[IrCl(COD)(IM

70). After exposu

d that a 105 ±

.4 T. The corre

plementary info

ction in polariza

s solvent and p

ancement of ove

er optimization

ssure of p-H2 a

oing to improve

gure 2. Effect of d

ell viability performed

of treatment with

olvent volume in the

and D2O - deuterium

P<0.005 and ns -

iable); one-way ANOVA.

arization levels

step should be ca

he toxicity of

opriate cell viab

reated with va

Figure 2, viab

ced when ethano

nd treated for a

me point (6 h),

ge in cell viab

, however, sig

eas, we found

(30:70) solutio

nt times when

(Figure 2 B an

ed that a significa

solvent conta

als to 50%. Th

similar (see SI

sis on SABRE

biocompatibilit

sure on cells

E using higher (

mal solvent

is 30% ethan

s of using this

MN. Thus, we p

Mes)], 20 mM

sure to 3 bar p

22 fold total si

esponding 1H N

ormation (see Fi

zation when com

polarization und

ver 2800-fold.[7]

n may be ach

and isotopically

ve the polarizatio

deuterated solven

ed on the indicated

h deuterated solv

he cell culture me

ium oxide. Data

- Not significant

OVA.

s it has been su

carried out in e

these solve

bility assay on

various dilutions

bility of both th

nol-d6 (100%) w

a short time (6

, 50% ethanol

bility. Extendin

gnificantly redu

that treatment

on did not show

compared to

nd see SI Figu

icant reduction

ained ethanol-d

The behavior of

SI Figure S2). Fr

solvents we co

ity it is importan

in culture (or

(>30%) ethanol

mixture for

nol-d6 in D2O

solvent directly

prepared a sam

d2-MN in eth

p-H2 at 298 K in

signal enhancem

NMR spectra a

Figure S3). Th

mpared to usin

nder of 3 bar p-7] As we have p

chieved through

y labelled catal

on levels.

ent mixture on ce

ed cell lines after (

olvents at different

edium was 10%. E

represent mean

vs. the untreated

suggested that t

ethanol-d6 prior

nt mixtures w

A549 and MC

s of ethanol-d6

he cell lines we

was added to c

6 h). Converse

l-d6 in D2O (1

ng the treatme

uced the viabi

of cells in a 30

w toxicity to ce

other deuterat

ure S1). Togeth

n in cell viability

d6 concentratio

analogous pro

rom these in vi

conclude that

nt to consider t

r in vivo) wh

l-d6 solvent.

biocompatibi

O, we wished

ly for the SABR

mple containing

hanol-d6 in D

n a 60 G field w

ment is observe

are shown in t

his is a significa

ng 100% ethan

-H2 gave a sign

previously show

h using a high

lysts and work

cell viability: MTT

(A) 6 h and (B) 24

ratios. The final

EtOD - ethanol-d

+ SD. *P<0.05,

ed control (100%

the

r to

we

CF7

6 in

ere

cell

sely,

1:1)

ent

ility

0%

cells

ted

her

y is

ons

otio

vitro

to

the

hen

ility

to

SABRE

g 5

D2O

we

ved

the

cant

nol-

nal

wn,

her

k is

T

4

al

d6

5,

%

FU

Ev

Ha

to

M

d2

se

so

co

ce

re

hig

lin

wh

th

tre

de

by

wh

To

in

ex

of

tra

F

c

t

of

v

m

s

of

c

FULL PAP

Evaluating the

aving found t

oxicity to cells i

MN dissolved in

2-MN with MN

selective deute

solvents. Importa

composition and

cells when trea

espectively). Ho

igher than 5 m

nes (Figure 3 C

hen compared

he viability of M

eated for long

euteration migh

y itself or that t

hen mixed in a

Together, our da

n an ethanol-d6

xposure on cel

f our cytotoxici

ansient effect

Figure 3. Cytotoxi

cell viability data

treated for (upper

of MN and d2-MN

volume in the cell

mean + SD and ar

significant differenc

of MN) is shown.

control group; one

PER

cytotoxicity of

that 30% etha

in vitro, we the

n this solvents m

to exclude an

eration in d2-

antly, d2-MN sh

d it did not redu

ated for up t

owever, longer

mM, are shown

C and 3 D). Sur

d to MN, d2-MN

MCF7 cells at

g time (24 h)

ht affect the tox

toxicity is more

alcohol solvent

ata suggest tha

:D2O solvent m

lls in culture. It

city assessment

these compou

xicity of alcohol

showing (A and

panel) 6 and (low

solubilized in 30%

ell culture medium

are from 3 independ

ences from untreat

n. *P<0.05 and ns

e-way ANOVA.

of substrate in

anol-d6 in D2O

en performed vi

mixture. Again

ny toxic effect

MN when diss

shows good sol

uce the viability

to 6 h (Figur

r treatment time

n to affect the

Surprisingly, as sh

N induced a sig

this concentra

). This further

xicity effects of

e pronounced a

t at this long tim

at the in vitro cy

mixture depend

It is worth men

nt is to allow us

unds or solve

solubilized meth

C) A549 and (B

(lower panel) 24 h w

30% ethanol-d6 in D2

edium was 10%. Data

endent experiment

reated control group

ns - Not significant

n solvent

O does not in

viability assay fo

here, we comp

that arise from

ssolved in al

lubility in this so

y of A549 and M

re 3 A and

es at concentra

viability of bot

shown in Figure

gnificant reduct

ration (5 mM)

r indicates tha

the substrate e

as an additive

me (24 h) expo

cytotoxicity of d

ds on the durat

ntioning that the

s to understan

vents play unde

methyl nicotinate: M

and D) MCF7 cell

with different dilutions

2O. The final solvent

a are presented

ents (n=3). Statistically

(or from protio for

ant vs. the untreat

3

nduce

or d2-

pared

m the

lcohol

solvent

MCF7

3 B,

ations

th cell

e 3 D,

ction in

when

at the

either

effect

osure.

d2-MN

tion of

e aim

nd the

er the

state

bioco

the p

actio

appro

comp

utility

Eval

Give

d6:D2

SABR

be ta

effect

lines.

For t

of [Ir

d6:D2

cells

activa

differ

treatm

of tim

when

medi

SABR

point

To

catal

solut

[IrCl(

solub

active

activa

(see

emul

resul

d2-M

treatm

delet

be du

SI Fi

volum

mixin

volum

these

reduct

to the

Th

the a

elimi

Method

We h

catal

SABR

deact

polar

postu

MTT

ells

ons

ent

as

ally

orm

ated

ed conditions.

compatibility wo

pharmacokinet

on of the tre

oach requires

paratively shor

y and further re

luating the bioc

en that the cyto

2O (30:70) is

BRE reaction (co

aken as a bolu

ct of the SABR

s.

this, we prepare

rCl(COD)(IMes)

2O (30:70) and

to various bo

vated mixture

rent time perio

ment with the

me (1 h) did no

n lowest volum

ium. However,

BRE reaction m

t.

To distinguish

lyst in the m

tions containin

(COD)(IMes)] a

ble in ethanol-d

ve catalyst wi

vated with H2.

SI Figure S4

lsion to treat ce

lts. Nevertheless

MN alone and

ments over 24

terious effect o

ue to the prese

Figure S5). In co

mes of the [Ir

ng of the preci

mes and short

se data indicate

ction in cell via

e presence of a

Thus, we concl

amount of act

inated.

hod of catalys

have therefore

lyst from soluti

BRE reaction pr

ctivating the

rization levels

ulated that th

We are awa

ould be depend

tics of the org

eated materia

the contrast a

rt time prior to

educes toxicity

ocompatibility

otoxic dosage o

well above th

considering only

us for treatmen

RE reaction mi

red a typical SA

s)] together wit

d activated it w

olus volumes (

and assessed

ods by MTT ass

SABRE reacti

ot reduce the vi

me (e.g. 1.25%

, cells that were

mixture showed

the cytotoxic

mixture we pre

ng various con

alone. Unfortun

d6:D2O (30:70)

ithout the pre

Under these co

S4) and cytoto

cells is not ex

ess, we treated

no adverse

4 h on both ce

of the SABRE

sence of the cata

contrast, the ce

rCl(COD)(IMes)

cipitate showed

rt time points

e that the SAB

ability at higher

activated cataly

clude that in or

ctivated catalyst

st deactivation

e developed a p

ion. The additi

revents reversi

SABRE pro

whilst extendi

he addition of

are that in an

dent on physiolo

ganism and th

al. Nonetheless

agent to stay i

o excretion as

concerns.

y of SABRE rea

of d2-MN in the

he amounts use

y less than or e

nt) we sought t

ixture on A549

SABRE solution

h 20 mM of d

with 3 bar H2.

(1.25, 2.5, 5 a

d the viability

ssay. As illustra

ion mixture ove

viability of A549

%) was added

e treated with

d less viability a

c effect of the

epared in pa

ncentrations of

nately, [IrCl(CO

) and moreove

esence of a

conditions, a p

oxicity assess

xpected to pro

d the cells with

effect was ob

cell lines. This in

reaction mixtu

alyst and not th

cells treated w

s)], prepared b

d loss of cell vi

(see SI Figur

ABRE reaction m

r volumes and

yst.

rder to achieve

st has to be e

on and removal

protocol to rem

ion of a chelat

sible exchange

ocess without

ing T1 relaxat

bathophenan

n in vivo sett

logical status a

he mechanism

ss, the SABR

in the body for

relaxation lim

action mixture

e solvent ethan

sed for a typica

equal to 10% w

to investigate t

9 and MCF7 c

containing 5 m

d2-MN in ethan

We exposed t

and 10%) of t

of the cells

rated in Figure

ver a short peri

9 and MCF7 ce

d to cell cultu

10% bolus of t

at the same tim

e substrate a

arallel analogo

either d2-MN

OD)(IMes)] is le

ver cannot form

substrate wh

precipitate form

ssment using th

ovide comparab

a solution of t

bserved even

ndicates that t

ure on cells cou

he substrate (se

ith various bol

by homogeneo

viability at high

re S6). Togeth

mixture induces

this is likely d

ve biocompatibi

either reduced

move the SABR

ting ligand to t

of the substra

t affecting t

ion times.[15] W

nthrolinedisulfo

ing

and

of

RE

r a

mits

e

nol-

cal

will

the

cell

mM

nol-

the

the

at

4,

iod

cells

ure

the

me

and

ous

or

ess

an

hen

med

his

ble

the

at

the

uld

see

lus

ous

her

her

s a

due

ility

or

RE

the

ate,

the

We

onic

FU

aci

irid

sc

SA

ad

re

formation of a new hydride species at δ to

Fi

as

hig

bi

FULL PAP

cid disodium sa

idium center

schematically in

SABRE reaction

ddition of a so

eaction by 1H

formation of a new hydride species at δ o [Ir(IMes)(BPS)

Figure S7). Afte

s eluent less th

igh mass reco

iocompatible et

Figure 4. Evalu

2.5, 5 and 10 %)

are from 3 indep

PER

salt (BPS) wou

whilst giving

n Figure 5. F

n mixture in et

solution of 2.0 e

H NMR spect

formation of a new hydride species at δ BPS)(d2-MN)(H)2]C

er filtration thro

han 2% of the

overy of d2-MN

ethanol-d6:D2O

uating the biocomp

%) of SABRE reaction

independent experiment

uld see it irreve

an opportuni

First, we prep

thanol-d6:D2O so

eq. of BPS in D

ctroscopy reve

formation of a new hydride species at δ −19.6 which we attribute Cl and confirm

ough DEAE-Se

e catalyst rema

N, which can

solvent mixtur

mpatibility of SAB

ion mixture and cell

ents (n=3). *P<0.05

versibly bind t

ity to remove

pared the acti

solution prior t

D2O. Followin

als the imme

−19.6 which we attribute m by LC-MS (se

Sephadex® with

ains in solution

be delivered i

res. This proto

BRE reaction mi

ell viability was as

05, **P<0.005 and

4

to the

ve the

resul

proce

ivated

to the

ng the

ediate

−19.6 which we attribute see SI

h D2O

n and

in the

ocol is

there

after

were

when

d6:D2

supp

deact

seco

attrib

mixture: (A) A549 and

assessed 1, 6 and 24

ns - Not significant

ltant species

cedure

efore efficient a

Importantly, th

the deactivatio

e 74 ± 21 fold

n compared to

2O (30:70).

plementary inf

ctivation and d

conds longer th

bute the loss to

and (B) MCF7 cell

24 h thereafter by

ant vs. the untreated

via ion-exch

at removing the

he hyperpolariz

on and depletio

which represe

o the standard1H NMR sp

formation (se

depletion proce

han a standa

o relaxation effe

ells were treated wit

by MTT assay. Data

ed control (100% v

change chrom

is

e SABRE cataly

zed SABRE sig

on steps. The t

sents a 30% re

rd SABRE sam

spectra are

see SI Figure

cess takes a

ard sample m

ects.

ith different bolus

a are presented as

viable); one-way AN

matography. T

show

yst from solutio

gnal is still visib

total signal gai

eduction in sign

mple in ethan

shown in t

e S7). As t

minimum of

measurement, w

volumes (0, 1.25,

s mean + SD and

ANOVA.

This

wn

on.

ble

ins

nal

nol -

the

the

12

we

,

and

FU

Cy

m

Th

th

10

by

pe

Pl

SA

fo

F

[

c

Sep

FULL PAP

Cytotoxicity a

mixture

The cytotoxicity

hen evaluated

0 %) of the reco

y following the

erformed with

Pleasingly, 10%

SABRE reaction

or up to 6 h of t

Figure 5. Schem

[IrCl(COD)(IMes)],

catalyst [Ir(IMes)(B

Sephadex® leads

Figure 6. Achi

various volumes

experiments (n=

Figure 7. Evaluvarious volumespresented as mean

PER

assessment of

of the catalyst

by taking diff

constituted mix

e treatment co

the non-que

% of the bolus

n mixture did n

treatment (Figu

ematic presentati

, d2-MN and H2 in

)(BPS)(d2-MN)(H)2

leads to less than 2% c

ieving biocompati

es of catalyst deplet

(n=3). *P<0.05, **P<

uating the biocompes (0, 2.5, 5, 7.5 and

ean + SD and are

of catalyst

st depleted sa

ferent volumes

xture in ethanol

conditions in the

enched SABRE

s containing th

not alter the via

ure 6 A). Simila

tation of the catal

n ethanol-d6:D2O

2]Cl as confirmed

catalyst contamina

atible SABRE by

pleted SABRE rea

P<0.005 and ns - Not

mpatibility of biphand 10%) of the bolu

from 3 independen

depleted SA

mples on cells

s (1.25, 2.5, 5

l-d6:D20 (30:70

e same mann

E reaction mi

he catalyst dep

ability of A549

arly, MCF7 cell

alyst deactivatio

(30:70) solution. 2

ed by1H NMR spe

ination.

y deactivating th

eaction mixture for

ot significant vs. the

hasic SABRE reacbolus from the aque

dent experiments (n=

5

ABRE

s was

5 and

0) and

ner as

ixture.

pleted

9 cells

l lines

show

volum

to 1

Howe

at bo

and 6

it is

meta

these

catal

mixtu

on procedure: 1.

2. Deactivation -

pectroscopy and

the catalyst: MTT

or 1, 6 and 24 h.

he untreated cont

eaction mixture: Mqueous fraction ofn=3).

wed no change

mes (10 %) of

h and for up to

ever, longer tre

oth lower and h

6 B). It is noted

unlikely to sho

abolic activity

se data indicate

lyst could over

ure when treati

1. Activation - The

Addition of BPS l

LC-MS. 3. Deple

T viability assay s

. Data are presen

ontrol (100% viable);

MTT viability assaya SABRE reaction

es in cell viab

the catalyst d

o 6 h with lowe

eatments (24 h

higher volume

d that when ex

ow the same lo

and detoxifica

e that the de

rcome the adve

ing live cells un

he active catalys

leads to the immedia

epletion - Ion-exchan

showing (A) A549

ented as mean +

able); one-way ANOVA

ay showing (A) A54ion in biphasic solv

bility when tre

depleted SABR

er volumes (≤ 5 %, Figure 6 B). h) showed sign

es in both cell

xtrapolating to a

ong-term toxici

cation mechan

eactivation and

verse effect of

nder the conditi

st is formed thro

ediate formation of

hange chromatograp

9 and (B) MCF7 c

SD and are from

OVA.

A549 and (B) MCF7olvents for 1, 6 and

eated with high

RE mixture for

≤ 5 %, Figure 6 B). nificant decrea

lines (Figure 6

an in vivo mod

city due to high

nisms. Togeth

d removal of t

SABRE reacti

ions used here

hrough reaction of

of the deactivated

ography on DEAE-

cells treated with

m 3 independent

cells treated withand 24 h. Data are

her

up

≤ 5 %, Figure 6 B). ase

6 A

del,

her

her

the

ion

e.

of

ed

-

hare

FU

Ac

W

ap

wa

m

dich

ph

be

ph

10

ap

th

C

To

th

fu

pr

te

4,

leve

va

sh

ch

wi

co

ef15N

ar

m

ef

pr

6

et

so

co

Th

re

ach

diso

DE

m

ho

po

ap

bi

de

de

be

is

st

ca

FULL PAP

chieving bioc

We hypothesize

pproach to SA

ay to deplete

method the SAB

ichloromethane

hase. Converse

etween the two

hase was isola

0%) were adde

ppropriate viab

he previous se

Conclusions

To conclude, we

hat it is possib

uture in vivo de

re-clinical/clinica

echnology. We

,6-d2-nicotinate

evels and long m

values across a

shown that deut

change in toxico

ill provide impo

contrast agents

ffect of other co

N) as we furthe

re interested

molecules that ca

The solvent

ffect on cell su

roduced adverse

h. We overca

thanol-d6:D2O

solvent deuterati

We have sh

contributor in red

Therefore, we h

emove it from

chieve this

isodium salt (B

DEAE-Sephadex

minimal adverse

ours exposure

olarization. Fu

pproach to

iocompatibility a

We are curre

elivery method

epletion or sep

e used in a clin

the main so

rategies to re

catalyst.

PER

ocompatible SA

zed that using

BRE catalysis

the solution

ABRE catalyst

e (DCM) phas

sely, the hyper

o. For toxicity a

ated and variou

ed to the cell c

bility assay at d

ections. Treatm

s

e have shown t

ble to create a

etection. This

cal disease st

have exempli

e (d2-MN) which

magnetic lifetim

a number of h

teration of the

cology when com

ortant informati

in the future. W

common isotopi

er expand our

in the tube

can sustain long

composition w

survival rate in a

se effects on th

ame this by di

(30:70) showe

ion had no effe

shown that the

ducing the viab

have develope

solution by io

we add ba

BPS) and subse

x®. The elue

e effects on a

and retained 7

urthermore, t

SABRE cat

after separatio

ently working t

d that includes

paration method

nical setting. Ad

source of toxicit

educe toxicity

ABRE by Bipha

g the recently

could be a m

of the iridium

st is located i

se and minima

rpolarized sub

assessment on

us bolus volume

culture medium

different time po

ment with the

through in vitro

a biocompatible

is an importan

state diagnosis

ified our toxicit

ch shows stron

mes. [7] By deter

human cancer

substrate did n

mpared to its p

ion for the deve

We will also lo

ic labelling stra

substrate profi

rculosis drug,

g lived singlet st

was shown to

an in vitro assa

he cells over ti

ilution with D2O

ed no cell deat

ect over their pr

e iridium cata

bility of cells in

ed simple and

on-exchange ch

athophenanthro

sequently flush t

ent from this

a number of ce

70% of the init

the recently

talysis also

on of the aqueo

towards develo

s all of the pro

ds to exclude t

Additionally, as

ty, we are co

through furth

asic catalysis

y reported bip

more rapid and

m catalyst. [16] In

in a chlorofor

ally in the aqu

bstrate is distri

n cells, the aqu

es (2.5, 5.0, 7.5

m. We performe

oints as illustra

e phase sepa

o cytotoxicity st

e SABRE bolu

nt step toward

using the SA

ty study for m

ng hyperpolariza

rmination of the

cell lines we

not show sign

protio analogue

velopment of SAB

ook to determin

ategies (e.g. 13C

ile. For exampl

, isoniazid [17],

states [4f, 18].

have a subst

ay. 100% etha

ime periods of

O and found t

th over 24 h. Ag

rotio analogues.

alyst is the la

the SABRE m

d robust metho

chromatograph

olinedisulfonic

the solution thr

procedure sh

cell lines for up

tial SABRE ind

reported bip

leads to

ous phase.

oping an autom

oposed deactiva

the catalyst tha

the SABRE ca

considering syn

her changes to

6

phasic

facile

n this

rm or

ueous

ibuted

ueous

.5 and

ed the

ated in

arated

SABR

MCF7

minim

when

resul

react

produ

the

appro

Figur

bipha

d6:D2

studies

us for

ds the

SABRE

methyl -

zation

he IC50

have

nificant

e. This

SABRE

ne the

C and

le, we

, and

stantial

anol-d6

up to

that a

Again

s.

argest

mixture.

hod to

hy. To

acid

rough

howed

p to 6

duced

phasic

bolus

mated

vation,

at can

catalyst

nthetic

o the

Exp

Chem

Aldric

oxide

follow

methy

Cell c

cance

by Pr

adeno

Prof.

were

penic

(all fr

humid

media

conflu

Tryps

Treat

cells

exclus

well

treatm

exper

subst

maxim

origin

otherw

the to

MTT

tetraz

determ

functi

metho

it is d

repres

were

above

(untre

the c

BRE mixture di

F7 cells at any

mal cytotoxic e

n treated with

lt indicates tha

ction mixture is

uce a biocomp

polarization le

oximately 2000

re S10). Never

asic mixture is

2O solution

xperimental

micals and reag

ch, Fisher or Alfa

e (D2O) and chlor

wing compounds

yl 4,6-d2 nicotina

culture. The hu

er cells (MCF7),

rof. Christoph Bo

ocarcinoma ce

Thomas Kaufm

grown in DMEM

cillin (100 U/ml),

from Gibco, Life

dified atmospher

a were changed

uence (90%) th

sin-EDTA (Invitro

tment of cells.

were counted

usion. The require

tissue culture p

tment so that they

riment. Before

trate, catalyst or

mum of 10 µl in

nally prepared.

wise indicated) w

total volume of the

assay. The

zolium bromide)

mine in vitro cy

tional mitochondr

od, MTT - tetrazo

directly proportion

esents a measure

For this assay,

taken in triplica

e) or kept as c

eated) in cell gro

cell growth med

id not reduce

y of the time

effect seen he

the substrate

at the cytotoxici

negated by thi

patible bolus by

evel achieved

0-fold for the sa

rtheless, we co

s higher than

under 3 b

Section

gents. All chem

fa-Aesar. Deutera

roform-d (CDCl3)

were prepared

ate[19] and [IrCl(CO

uman alveolar ad

, cervical cancer

orner, IMMZ, Fre

ell line MDA

fmann, University

M supplemented w

streptomycin (10

fe Technologies).

re under standar

at regular interv

the cells were p

ogen) solution.

Immediately aft

in a haemocyto

red number of ce

plate (Nunc)) w

y are in exponen

treatment vario

r the SABRE re

the same solven

For cell assay

we kept the final

e cell growth med

MTT (3-(4,5-Di

assay is a com

ytotoxicity of the

rial dehydrogena

olium is reduced

nal to the numbe

e for cell viability

cells (normally 1

ates and treated

cell viability con

owth media. Afte

dia was replace

the viability o

points tested

ere is similar to

alone (see SI

city associated

is method. Whi

y this phase se

by the bipha

same substrate

conclude that p

that observed

bar p-H2 an

micals were purch

ated solvents (eth

)) were purchase

according to lite

OD)(IMes)].[20]

denocarcinoma c

r cells (HeLa) we

eiburg, Germany.

A-MB-231 was

of Bern, Switze

with 10% fetal bo

00 µg/ml) and L

. The cells were

rd conditions (37

vals and upon re

passaged after

fter Trypsin-EDT

tometer by Try

ells (normally 10

was seeded 24

ntial growth phas

ous volumes o

eaction mixture

nt from which the

ys, throughout t

amount of the so

dium (i.e. 10 µl in

imethyl-2-thiazoly

mmonly used colo

e given compoun

ase activity in liv

d to insoluble form

er living cells. Th

as well.

104 cells per wel

d with a range o

ntrol with no co

ter incubation at d

ed with fresh m

of either A549

(Figure 7). T

o the effect se

Figure S9). Th

with the SABR

ile we are able

eparation metho

asic catalysis

(d2-MN), (see

polarization in t

d in an ethan

nd at 298

hased from Sigm

hanol-d6, deuteri

ed from Sigma. T

erature procedur

cells (A549), bre

ere kindly provid

. The human bre

s a gift fr

erland. All the ce

ovine serum (FB

L-Glutamine (2 m

re maintained in

7°C; 5% CO2). T

eaching appropri

brief exposure

TA treatment via

ypan blue (Sigm

04 per well in a

hours (h) befo

se at the start of t

of the bolus fro

were diluted to

the compounds w

this study (unle

olvent to be 10%

n 100 µl).

yl)-2,5-diphenyl-2

orimetric method

nd by means o

ving cells [12]. In t

mazan crystals a

herefore, this ass

ll in a 96-well pla

of compounds (s

ompound treatme

desired time poi

media (100 µl) a

or

The

seen

This

SABRE

e to

od,

is

e SI

the

nol -

K.

ma-

ium

The

res:

east

ded

east

from

ells

BS),

mM)

n a

The

iate

to

able

ma)

96-

fore

the

from

to a

were

ess

% in

2H-

d to

of a

this

and

say

ate)

see

tment

ints

and

FU

inc

filte

me

cry

ab

nm

ab

DM

(i.e

of

the

Ea

an

Ev

dis

co

mi

MT

ca

Ca

mM

3.0

of

ba

2.0

elu

tre

eth

fur

Bip

[Ir

(1

an

ba

tre

ab

St

(n

the

an

(A

se

A

Th

fu

wi

C

Th

Ke

ca

R

FULL PAP

cubated with 10

ter (0.22 µm) ste

edium with MTT

ystals were solu

bsorbance of the

m using a mic

bsorbance value

MSO and the ab

.e. untreated) wa

the compound t

e untreated cont

ach assay was

nalysis of the data

valuation of IC

ssolved in cell

oncentrations of th

inimum of 1.25 m

TT assay after 4

alculated by using

atalyst deactiva

M) and d2-MN (2

.0 mL) was degas

3 bar. After 5 m

athophenanthrolin

.0 eq.) in water

uted through D

eatment on cells

thanol-d6 in D2O.

rther diluted as in

iphasic SABR

rCl(COD)(IMes)]

1.5 mL) and D2O

nd was degassed

ar. After phase se

eatment various

bove.

tatistical analys

n=3) and all data

e mean. The sig

nd the untreated

ANOVA) using G

et at P values <0.0

Acknowledg

The Wellcome

unding. We tha

ider CHyM tea

Conflict of in

The authors decl

eywords: SABR

atalysis • cytotoxi

References:

PER

µl of MTT (5 mg/

terilized) for 4 h in a humidified atmosphere at 37°C. The was then carefu

ubilized in 100 µ

e coloured (purpl

croplate reader

es (averaged ou

bsorbance of cell

as taken as 100 %

treated samples

rol sample and a

repeated for a

a.

IC50. For the e

growth media

the compound (ra

mM) in ½ serial d

48 h of treatmen

g Graphpad Priz

ation and remov

20 mM, 4.0 eq.)

ssed prior to the

minutes, the samp

nedisulfonic acid

(1.0 mL) was a

DEAE-Sephadex®

the eluent was v

. Prior to treatme

ndicated above.

RE - Biphas

(5 mM) and d2-

O (containing 0.9

d prior to the intro

separation the aq

volumes of the b

sis. All experimen

is presented as

gnificance of the

control was asse

GraphPad Prizm

.05.

gements

Trust (092506

ank Graham C

m for scientific

interest

clare no conflict

RE • hyperpola

icity

:

g/ml, dissolved in

 h in a humidified atmosphere at 37°C. The ully aspirated and

µl of dimethyl s

le) solution was

r (MultiskanGO,

ut of triplicates)

ls exposed to ce

% viable (i.e. co

were then calcul

are normally expr

minimum of thre

evaluation of IC

and cells were

ranging from a m

dilutions. Cell via

nt. IC50 values of

m software.

val. A solution o

in ethanol-d6:D2

e introduction of h

ple was opened t

d disodium salt h

added. The resu

® (2.5 g) with

vacuum dried an

ent various volum

sic separatio

-MN (20 mM, 4.0

9% NaCl) (1.5 m

oduction of hydro

queous layer was

bolus were furth

nts were perform

s mean and stan

e differences betw

essed by one-wa

Software. Statis

6 and 098335

Cheetham, Ga

c discussions.

ct of interest.

arization • bioco

n cell growth med

 h in a humidified atmosphere at 37°C. The d the formed form

sulfoxide (DMSO

then measured a

, Thermofisher)

were blanked a

ell growth medium

ntrol). The cell v

lated by normalis

ressed as % of c

ee times for sta

IC50, compounds

e treated with v

maximum of 80 m

ability was analys

of MN and d2-MN

of [IrCl(COD)(IMe

2O (30:70, total v

hydrogen at a pre

to air and a solu

hydrate (BPS) (1

ulting suspensio

water (11 mL

nd reconstituted i

mes of the bolus

n. A solutio

.0 eq.) in chlorof

ml) was mixed to

ogen at a pressu

s removed and p

her diluted as ind

med at least three

ndard deviation (

tween treated sa

ay analysis of va

stical significanc

5) are thanke

ary Green and

ompatibility • bip

7

dia and

 h in a humidified atmosphere at 37°C. The mazan

O). The

at 570

. The

against

m only

viability

sing to

control.

tatistical

s were

various

mM to a

sed by

N were

IMes)] (5

volume

essure

ution of

10 mM,

on was

L). For

in 30%

s were

on of

oform-d

together

ure of 3

prior to

dicated

e times

(SD) of

amples

ariance

ce was

ked for

d the

iphasic

[1]

[2]

[3]

[4]

[5]

[6]

[7]

[8]

[9]

[10]

[11]

[12]

[13]

[14]

(a) J. Kurhanew

Comment, C. H.

Leach, S. S. Raja

Neoplasia 2011

Johannesen, A.

A. Kjar, America

2015, 5, 548-560

(a) P. Nikolaou,

Newton, S. Barc

Rosen, S. Patz

Proceedings of t

of America 2013

Walkup, B. M.

Dabaghyan, K. R

E. Y. Chekmene

32, 541-550.

K. D. Atkinson,

Green, J. López

Chemical Societ

(a) T. Theis, G. X

Huhn, V. Blum,

Warren, Science

Coffey, R. V. Sh

Warren, E. Y. C

2015, 137, 1404

Chemical Comm

Rayner, G. G. R

Journal of Physic

A. Burt, P. J. Ra

Duckett, Chemic

Roy, P. Norcott,

– A European Jo

Shchepin, C. P.

S. Warren, E. Y.

M. J. Cowley, R

Duckett, G. G. R

R. E. Mewis, Jo

6134-6137.

(a) R. W. Adams

Green, Journal

Aspers, B. J. A

Wijmenga, M. Te

66.

P. J. Rayner, M.

Green, L. A. R. H

National Academ

114, E3188-E31

(a) B. Belleau, J

102-104; (b) T. G

A. Bergman, U.

17.

K. S. Egorova, V

(a) L. C. Jumbelic

2006, 19, 147-15

Contact Dermati

T. Mosmann, J I

L. S. Lloyd, A. A

Cowley, G. J. De

R. Highton, A. J

Mewis, A. D.

Technology 2014

(a) M. Fekete, C.

D. Roberts, F. C

7870-7880; (b) P

K. J. Hermkens,

Tinnemans, M. T

Chemistry – A

Colell, M. Emond

X. Ortiz, P. Span

wicz, D. B. Vignero

. Cunningham, R.

jan, R. R. Rizi, B.

1, 13, 81-97; (b

E. Clemmensen,

an journal of nuc

0.

A. M. Coffey, L. L

rcus, I. Muradyan,

z, M. J. Barlow,

the National Acad

3, 110, 14150-141

. Gust, N. Whit

Ranta, G. D. Moro

ev, B. M. Goodson

M. J. Cowley, P.

z-Serrano, A. C.

ty 2009, 131, 1336

X. Ortiz , A. W. J.

S. J. Malcolmson

e Advances 2016

hchepin, K. W. W

Chekmenev, Journ

4-1407; (c) V. V. Z

munications 2015,

R. Green, L. A. R.

ical Chemistry B 2

ayner, S. J. Hart, A

ical Communicatio

P. J. Rayner, G. G

ournal 2017, 23,

N. Tanner, J. F. P

. Chekmenev, Che

R. W. Adams, K.

R. Green, J. A. B.

ournal of the Am

s, S. B. Duckett, R

of Chemical Ph

A. van Weerdenb

essari, Journal of

. J. Burns, A. M. O

Highton, R. E. Me

my of Sciences o

194.

J. Burba, M. Pinde

G. Gant, J Med Ch

Svedberg, E. Nils

V. P. Ananikov, Org

lic, F. T. Liebel, M

52; (b) N. Issacha

itis 1998, 39, 182-

Immunol Methods

Asghar, M. J. Bu

Dear, S. B. Duckett

J. J. Hooper, M. K

Roberts, A. J.

14, 4, 3544-3554.

. Gibard, G. J. Dea

Cisnetti, S. B. Du

P. Spannring, I. Re

, N. G. J. van der Z

Tessari, B. Blümi

European Journa

dts, A. W. J. Loga

nnring, Q. Wang,

ron, K. Brindle, E

. J. DeBerardinis,

. D. Ross, W. S. W

b) H. Gutte, A.

J. H. Ardenkjar-La

clear medicine and

L. Walkup, B. M.

, M. Dabaghyan,

E. Y. Chekmene

demy of Sciences

155; (b) P. Nikolao

iting, H. Newton

oz, M. S. Rosen, S

n, Magnetic Reson

. I. P. Elliott, S. B

Whitwood, Journ

62-13368.

. Logan, K. E. Cla

n, E. Y. Chekmen

6, 2; (b) T. Theis,

Waddell, F. Shi, B.

rnal of the America

Zhivonitko, I. V. Sk

, 51, 2506-2509; (d

. Highton, R. E. M

2015, 119, 5020-5

A. C. Whitwood, G

ions 2016, 52, 14

G. R. Green, S. B

10496-10500; (g)

P. Colell, B. M. Go

emPhysChem 201

D. Atkinson, M. C

. Lohman, R. Kers

merican Chemical

R. A. Green, D. C.

hysics 2009, 131

burg, M. C. Feite

f Magnetic Reson

Olaru, P. Norcott,

ewis, S. B. Duckett

of the United State

ell, J. Reiffenstein

hem 2014, 57, 359

sson, Contact Derm

rganometallics 201

M. D. Southall, Skin

ar, Y. Gall, M. T. B

-186.

s 1983, 65, 55-63.

urns, A. Charlton,

t, G. R. Genov, G

Khan, I. G. Khaza

Ruddlesden, C

ar, G. G. R. Green

uckett, Dalton Tra

eile, M. Emondts,

Zwaluw, B. J. A. v

ich, F. P. J. T. R

al 2016, 22, 927

an, K. Shen, J. Ba

S. J. Malcolmson

E. Y. Chekmenev,

, G. G. Green, M.

Warren, C. R. Mallo

E. Hansen, H.

Larsen, C. H. Niels

d molecular imag

Gust, N. Whiting,

G. D. Moroz, M.

ev, B. M. Goods

of the United Sta

ou, A. M. Coffey, L

n, I. Muradyan,

S. Patz, M. J. Barlo

nance Imaging 20

B. Duckett, G. G.

rnal of the Americ

laytor, Y. Feng, W.

nev, Q. Wang, W.

M. L. Truong, A.

. M. Goodson, W

an Chemical Socie

kovpin, I. V. Kopty

(d) M. J. Burns, P

Mewis, S. B. Duck

5027; (e) A. M. Ola

G. G. R. Green, S.

4482-14485; (f) S.

B. Duckett, Chemis

) D. A. Barskiy, R

oodson, T. Theis,

17, 18, 1493-1498

C. R. Cockett, S.

rssebaum, D. Kilgo

Society 2011, 1

Williamson, G. G

1; (b) N. Eshuis,

ers, F. Rutjes, S.

nance 2016, 265,

, M. Fekete, G. G.

t, Proceedings of

es of America 20

in, Science 1961, 1

95-3611.

rmatitis 1995, 32,

17, 36, 4071-4090

in Pharmacol Phy

Borrel, M. C. Poelm

, S. Coombes, M

G. G. R. Green, L.

al, R. J. Lewis, R.

Catalysis Science

n, A. J. J. Hooper,

ransactions 2015,

P. P. M. Schleker,

van Weerdenburg

Rutjes, M. C. Feite

77-9282; (c) J. F.

ae, R. V. Shchepin

n, E. Y. Chekmen

, A.

. O.

lloy,

H.

lsen,

ging

, H.

. S.

son,

ates

L. L.

M.

ow,

014,

. R.

can

. P.

. S.

. M.

. S.

ciety

yug,

P. J.

kett,

laru,

. B.

. S.

istry

R. V.

W.

8.

. B.

our,

133,

. R.

, R.

. S.

59-

. R.

the

017,

133,

14-

0.

ysiol

lman,

M. J.

. A.

. E.

e &

r, A.

44,

r, N.

rg, P.

ers,

. P.

in, G.

nev,

FU

[15

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5] R. E. Mewis,

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7] P. Norcott, P

European Jo

8] S. S. Roy, P.

Chem Int Ed

9] P. J. Rayner,

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s, F. P. J. T. Rutje

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, M. Fekete, G. G.

ommunications 20

M. Olaru, G. G. R

ournal, DOI: 10.100

P. J. Rayner, G. G

ournal, n/a-n/a.

P. Norcott, P. J. Ra

Engl 2016, 55, 15

r, M. J. Burns, A. M

R. Highton, R. E.

ademy of Sciences

quez-Serrano, B.

tions 2002, 2518-2

jes, B. Blümich, T

mical Society 2017

. R. Green, A. C. W

015, 51, 9857-9859

R. Green, S. B. D

002/chem.2017027

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ayner, G. G. Gree

5642-15645.

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. Mewis, S. B. Duc

s 2017, 114, E318

. T. Owens, J.

2519.

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7, 139, 7761-7767

Whitwood, S. B. D

9.

Duckett, Chemist

716.

. Duckett, Chemis

en, S. B. Duckett,

rcott, M. Fekete, G

ckett, Proceedings

88-E3194.

M. Buriak, Ch

8

Warren,

7.

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istry – A

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