S1
Supplementary
Materials
A Family of Rhodium Complexes
with Selective Toxicity towards
Mismatch Repair
-
Deficient Cancers
Kelsey M. Boyle and Jacqueline K. Barton*
Division of Chemistry and Chemical Engineering
California Institute of Technology
Pasadena
CA, 91125
S
2
Additional
Methods:
Synthesis of [Rh(L)(chrysi)(PPO)]
2+
Complexes
For L = bpy
RhCl
3
3H
2
O (270
mg, 1.0 mmol, 1 equiv.) and KCl (78
mg, 1.0 mmol, 1 equiv.) were refluxed
in methanol (8 mL) for 2 hours at 98
°C. 2,2’
-
bipyridine (bpy, 160 mg, 1.0 mmol, 1 equiv)
was added in a minimum volume of methanol and refluxed for 4
h
, during which the deep
red solution turned to golden precipitate. The solution was filtered over a medium
fritted
filter
and rinsed with met
hanol and dried under vacuum (380
mg, 84% crude yield)
[Rh(bpy)Cl
4
]K (380
mg, 0.86 mmol, 1 equiv.) was added to an oven
-
dried 25
mL Schlenk
flask and degassed under argon. Neat triflic acid (10
g, excess) was added to the flask under
positive Ar pressure.
The solution turned deep red upon triflic acid addition. The flask was
purged to remove HCl gas and stirred for 12
h
. The solution was then added dropwise to
300
mL cold, stirring ether at
-
78
°C to produce a yellow
-
brown precipitate. The precipitate
was
filtered over a medium frit, washed with cold ether, and dried under vacuum.
[Rh(bpy)(OTf)
4
]K was combined with NH
4
OH (28% w/v, 40
mL, excess) and stirred at 40
°C for 1
h
, during which the solution became a foggy light yellow. The solvent was removed
unde
r vacuum (280
mg, 42% crude yield).
[Rh(bpy)(NH
3
)
4
](OTf)
3
(280
mg, 0.36 mmol, 1 equiv.) was combined with 5,6
-
chrysene
-
quinone (100
mg, 0.39 mmol, 1 equiv.) and 9:1 MeCN:H
2
O (40
mL) and NaOH (1
M, 2
mL)
and stirred for 1
h
. The solution changed from the bright orange of the
free
ligand to a red
-
brown solution with no precipitate. The reaction was quenched with HCl (1
M, 2
mL),
producing an even deeper red solution, and the solvent was removed under vacuum. The
red product w
as purified over a C18 SepPak, pre
-
equilibrated with 0.1%TFA (aq) and
eluted with 25% MeCN, 75% of 0.1% TFA (aq). (100
mg, 33% yield)
S
3
[Rh(bpy)(chrysi)(NH
3
)
2
](TFA)
3
(100
mg, 0.12 mmol, 1 equiv.) was combined with PPO (23
mg, 0.17 mmol, 1.4 equiv.) in 1:1 E
tOH:H
2
O (10 mL) and refluxed 12
h
. The solvent was
removed under vacuum and the product was purified by HPLC (85:15 MeCN:0.1% TFA (aq)
to 95:5 MeCN:0.1% TFA (aq) over 30
min
). The purified product was converted to the
chloride salt using Sephadex QAE resin
charged with MgCl
2
. (24 mg, 30% purified yield).
For L = HDPA
RhCl
3
3H
2
O (1.0 g, 3.8 mmol, 1 equiv.) was refluxed in concentrated HCl (38% w/v, 30
mL)
for 3
h
at 98
°C. 2,2’
-
dipyridylamine (HDPA, 1.3 g, 7.6 mmol, 2 equiv
) was added in a
minimum volume of HCl, followed immediately by boiling water (200 mL). The solution
was refluxed for 12
h
, then cooled to 4
°C. The golden precipitate was filtered over a
Buchner funnel and dried under vacuum. (2.2 g, >100% crude yield)
[
Rh(HDPA)Cl
4
][H
3
O] (2.2 g, 1 equiv.) was added to an oven
-
dried 25
mL Schlenk flask and
degassed under argon. Neat triflic acid (10
g, excess) was added to the flask under positive
Ar pressure. The solution turned deep red upon triflic
acid addition. The flask was purged
to remove HCl gas and stirred for 12
h
. The solution was then added dropwise to 200
mL
cold, stirring ether at
-
78
°C to produce a yellow
-
brown precipitate. The precipitate was
filtered over a medium
fritted filter
, was
hed with col
d ether, and dried under vacuum.
[Rh(HDPA)(OTf)
4
][H
3
O] was combined with NH
4
OH (28% w/v, 100
mL, excess) and stirred
at 40
°C for 45
min
, during which the solution became a foggy light yellow. The solvent was
removed under vacuum and the produ
ct was dissolved in a minimal amount of water and
precipitated with 10:1 ether:EtOH, filtered over a medium frit
ted filter
, and dried further
under vacuum. (400
mg, 10% crude yield).
[Rh(HDPA)(NH
3
)
4
](OTf)
3
(400
mg, 0.51 mmol, 1 equiv.) was combined with
5,6
-
chrysene
-
quinone (140mg, 0.55 mmol, 1 equiv.) and MeCN (65
mL) and NaOH (1
M, 8
mL) and
stirred for 12
h
. The solution changed from the bright orange of the
free
ligand to a red
-
brown solution with no precipitate. The reaction was quenched with HCl (1
M, 8
mL),
S
4
producing an even deeper red solution, and the solvent was removed under vacuum. The
red product was purified over a C18 SepPak, pre
-
equilibrated with 0.1%TFA (aq) and
eluted with 25% MeCN, 75% of 0.1% TFA (aq). (220
mg, 51% yield)
[Rh(HDPA)(chr
ysi)(NH
3
)
2
](TFA)
3
(70
mg, 0.08 mmol, 1 equiv.) was combined with PPO (24
mg, 0.17 mmol, 2.1 equiv.) in 1:1 EtOH:H
2
O (20 mL) and refluxed 7
d
. The solvent was
removed under vacuum and the product was purified by HPLC (85:15 MeCN:0.1% TFA (aq)
to 95:5
MeCN:0.1% TFA (aq) over 30
min
). The purified product was converted to the
chloride salt using Sephadex QAE resin charged with MgCl
2
. (6 mg, 10% purified yield).
For L = 4,7
-
DMP
RhCl
3
3H
2
O (500
mg, 1.9 mmol, 1 equiv.) and KCl (150
mg, 2.0 mmol, 1 equiv.)
were
refluxed in methanol (10 mL) for 2
h
at 98
°C. 4,7
-
dimethyl
-
1,10
-
phenanthroline (
4,7
-
D
MP,
400 mg, 1.9 mmol, 1 equiv) was added in a minimum volume of methanol and refluxed for
4
h
, during which the deep red solution turned to golden precipitate. The
solution was
filtered over a medium frit
ted filter
and rinsed with methanol and dried under vacuum
(800
mg, 86% crude yield)
[Rh(4,7
-
DMP)Cl
4
]K (800
mg, 1.6 mmol, 1 equiv.) was added to an oven
-
dried 25
mL
Schlenk flask and degassed under argon. Neat trifl
ic acid (10
g, excess) was added to the
flask under positive Ar pressure. The solution turned deep red upon triflic acid addition.
The flask was purged to remove HCl gas and stirred for 12
h
. The solution was then added
dropwise to 250
mL cold, stirring et
her at
-
78
°C to produce a yellow
-
brown precipitate.
The precipitate was filtered over a medium frit, washed with cold ether. The product,
[Rh(4,7
-
DMP)(OTf)
4
]K was combined with NH
4
OH (28% w/v, 50
mL, excess) and stirred at
40
°C for 1
h
, during which the solution became a foggy brown. The solvent was removed
under vacuum and the product was suspended in EtOH (5
mL), filtered over a medium
frit
ted filter
, and
rinsed with cold ethanol, and d
ried further under vacuum. (200
mg, 15%
crude yie
ld).
S
5
[Rh(4,7
-
DMP)(NH
3
)
4
](OTf)
3
(200
mg, 0.24 mmol, 1 equiv.) was combined with 5,6
-
chrysene
-
quinone (70mg, 0.39 mmol, 1.6 equiv.) and 6:1 MeCN:H
2
O (35
mL) and NaOH (1
M, 5
mL) and stirred for 1
h
. The solution changed from the bright orange of the
free
l
igand
to a green
-
brown solution with no precipitate. The reaction was quenched with HCl (1
M, 5
mL), producing a deep red solution, and the solvent was removed under vacuum. The red
product was HPLC purified (85:15 MeCN:0.1% TFA (aq) to 95:5 MeCN:0.1% TFA
(aq) over
30
min
). (100
mg, 46% purified yield)
[Rh(4,7
-
DMP)(chrysi)(NH
3
)
2
](TFA)
3
(50
mg, 0.03 mmol, 1 equiv.) was combined with PPO
(9
mg, 0.07 mmol, 2 equiv.) in 1:1 EtOH:H
2
O (10 mL) and refluxed 12
h
. The solvent was
removed under vacuum and the produc
t was purified by HPLC (85:15 MeCN:0.1% TFA (aq)
to 95:5 MeCN:0.1% TFA (aq) over 30
min
). The purified product was converted to the
chloride salt using Sephadex QAE resin charged with MgCl
2
. (4 mg, 10% purified yield).
For L = 5,6
-
DMP
RhCl
3
3H
2
O (1.0 g, 3.8 mmol, 1 equiv.) and KCl (290 mg, 3.9 mmol, 1 equiv.) were refluxed
in methanol (15 mL) for 2
h
at 98
°C. 5,6
-
dimethyl
-
1,10
-
phenanthroline (5,6
-
DMP, 790 mg,
3.8 mmol, 1 equiv) was added in a minimum volume of methanol and refluxed for 4
h
,
dur
ing which the deep red solution turned to beige precipitate. The solution was filtered
over a medium frit and rinsed with methanol and dried under vacuum (1.7 g, 91% crude
yield)
[Rh(5,6
-
DMP)Cl
4
]K (1.7 g, 3.4 mmol, 1 equiv.) was added to an oven
-
dried 25
mL Schlenk
flask and degassed under argon. Neat triflic acid (10
g, excess) was added to the flask under
positive Ar pressure. The solution turned deep red upon triflic acid addition. The flask was
purged to remove HCl gas and stirred for 12
h
. The solutio
n was then added dropwise to
200
mL cold, stirring ether at
-
78
°C to produce a beige precipitate. The precipitate was
filtered over a medium frit
ted filter
, washed with cold ether. The product, [Rh(5,6
-
DMP)(OTf)
4
]K was combined with NH
4
OH (28% w/v, 100
mL
, excess) and stirred at 40
°C
for 40
min
. The solvent was removed under vacuum and the product was dissolved in
S
6
minimal EtOH and precipitated in ether, filtered over a medium frit
ted filter
, and
d
ried
further under vacuum. (2.2 g, 77% crude yield).
[Rh(
5,6
-
DMP)(NH
3
)
4
](OTf)
3
(830 mg, 1.0 mmol, 1 equiv.) was combined with 5,6
-
chrysene
-
quinone (250
mg, 1.0 mmol, 1 equiv.) and 11:1 MeCN:H
2
O (250
mL) and NaOH (1M, 4
mL)
and stirred for 1
h
. The solution changed from the bright orange of the
free
ligand to a
green
-
brown solution with no precipitate. The reaction was quenched with HCl (1
M, 4
mL), producing a deep red solution, and the solvent was removed under vacuum. The red
product was HPLC purified (85:15 MeCN:0.1% TFA (aq) to 95:5 MeCN:0.1% TF
A (aq) over
30
min
). (540 mg, 62% purified yield)
[Rh(5,6
-
DMP)(chrysi)(NH
3
)
2
](TFA)
3
(40mg,
0.04
mmol, 1 equiv.) was combined
with PPO
(11 mg, 0.08
mmol, 2 equiv.) in 1:1 EtOH:H
2
O (10 mL) and refluxed 12
h
. The solvent was
removed under vacuum and the pro
duct was purified by HPLC (85:15 MeCN:0.1% TFA (aq)
to 95:5 MeCN:0.1% TFA (aq) over 30
min
). The purified product was converted to the
chloride salt using Dowex 1x2 500
-
100 mesh ion exchange resin. (8 mg, 23% purified
yield).
For L = DIP
RhCl
3
3H
2
O (770
m
g, 2.9 mmol, 1 equiv.) and KCl (230 mg, 3.1 mmol, 1 equiv.) were
refluxed in methanol (15 mL) for 2 hours at 98
°C. 4,7
-
diphenyl
-
1,10
-
phenanthroline (DIP,
970 mg, 2.9 mmol, 1 equiv) was added in a minimum volume of methanol and refluxed for
4
h
, du
ring which the deep red solution turned to beige
-
yellow precipitate. The solution
was filtered over a medium frit and rinsed with methanol and dried under vacuum (1.7 g,
95% crude yield)
[Rh(DIP)Cl
4
]K (1.7 g, 2.8 mmol, 1 equiv.) was added to an oven
-
dried
25
mL Schlenk flask
and degassed under argon. Neat triflic acid (10
g, excess) was added to the flask under
positive Ar pressure. The solution turned deep red upon triflic acid addition. The flask was
S
7
purged to remove HCl gas and stirred for 12
h
. The sol
ution was then added dropwise to
200
mL cold, stirring ether at
-
78
°C to produce a beige precipitate. The precipitate was
filtered over a medium frit
ted filter
, washed with cold ether. The product, [Rh(DIP)(OTf)
4
]K
was combined with NH
4
OH (28% w/v, 100
mL
, excess) and stirred at 40
°C for 40
min
. The
solvent was removed under vacuum and the product was dissolved in minimal EtOH and
precipitated in ether, filtered over a medium frit
ted filter
, and d
ried further under vacuum.
(1.9 g, 72% crude yield).
[Rh(
DIP)(NH
3
)
4
](OTf)
3
(510 mg, 0.54 mmol, 1 equiv.) was combined with 5,6
-
chrysene
-
quinone (140
mg, 0.55 mmol, 1 equiv.) and 11:1 MeCN:H
2
O (250
mL) and NaOH (1
M, 4
mL) and stirred for 1
h
. The solution changed from the bright orange of the
free
ligand to a
gr
een
-
brown solution with no precipitate. The reaction was quenched with HCl (1
M, 4
mL), producing a deep red solution, and the solvent was removed under vacuum. The red
product was purified over a C18 SepPak, pre
-
equilibrated with 0.1%TFA (aq
) and eluted
with 25% MeCN, 75% of 0.1% TFA (aq). (620
mg, >100% crude yield)
[Rh(DIP)(chrysi)(NH
3
)
2
](TFA)
3
(52
mg, 0.05 mmol, 1 equiv.) was combined with PPO (15
mg, 0.11 mmol, 2 equiv.) in 9:1 EtOH:H
2
O (10 mL) and refluxed 12
h
. The solvent was
removed
under vacuum and the product was purified by HPLC (85:15 MeCN:0.1% TFA (aq)
to 95:5 MeCN:0.1% TFA (aq) over 30
min
). The purified product was converted to the
chloride salt using Dowex 1x2 500
-
100 mesh ion exchange resin. (15 mg, 33% yield).
Radiolabelin
g of DNA
DNA was purchased from IDT DNA and purified by HPLC on a C18 reverse
-
phase column.
The DNA was quantified using extinction coefficients provided by IDT DNA.
A DNA hairpin
(5'
-
GGCAGG
X
ATGGCTTTTTGCCAT
Y
CCTGCC
-
3
'
, where XY=CG or CC for a well
-
matched
or
mismatched hairpin, respectively) was incubated with γ
-
32
P ATP and polynucleotide kinase
at 37
̊
C for 2.5
h
, then purified using two BioRad
Micro Bio
-
Spin 6 columns following the
manufacturer’s instructions. Solvent was removed from the DNA, and the DNA was
dissolved in 10 mM NaP
i
, pH 7.1. A 2 μM solution of DNA was made in 100 mM NaCl and 20
S
8
mM NaP
i
buffer containing approximately 1%
32
P
-
lab
eled DNA and 99% unlabeled DNA. To
anneal, the DNA was heated on a 90 ̊C heat block for 10
min
, cooled to room temperature
over the course of 2.5
h
, and then stored at 4
̊
C prior to use.
Table S1: Converted n
uclear rhodium content
values
Compound
ng [Rh] / mg
[nuclear
protein]
ng [Rh] /
nuclei × 10
-
8 a
[Rh] μM
b
Increase
over
extracellular [Rh]
c
N Cells
PHEN
3.0
10.0
3.6
7.2
BPY
1.8
5.8
2.1
4.2
HDPA
6.5
21.3
7.7
15.5
4,7DMP
4.1
13.3
4.8
9.6
5,6DMP
4.1
13.5
4.9
9.8
DIP
8.0
26.2
9.5
47.5
RhBC
26.6
87.2
31.6
3.2
O Cells
PHEN
2.6
8.6
3.1
6.3
BPY
3.5
11.6
4.2
8.4
HDPA
7.3
23.8
8.6
17.2
4,7DMP
5.0
16.4
5.9
11.9
5,6DMP
6.8
22.4
8.1
16.2
DIP
11.6
38.0
13.7
68.7
RhBC
37.3
122.2
44.3
4.4
a
a conversion factor of
3.28 × 10
−
8
mg nuclear protein / nucl
ei was determined in
reference
1
b
the nucleus was approximated as a sphere of radius 4
μm following reference
2
c
Increase taken as
ratio between the nuclear rhodium concentration and the dosing
concentration of each
metalloinsertor.
S
9
Figure S1
:
(A)
HPLC trace of the purified Δ
-
and Λ
-
enantiomers of
[Rh(phen)(chrysi)(PPO)]
2+
, (B) a
CD spectrum showing the enantiomeric nature
of
the two
samples, and (C) a CD spectrum taken after one month of incubation at standard
temperature and pressure, showing no racemization.
Figure S2
:
MTT assays of Δ
-
and Λ
-
[Rh(phen)(chrysi)(PPO)]Cl
2
with HCT116N (MMR
proficient) and HCT116O (MMR de
ficient
) cell lines. Both enantiomers show selective
cytotoxicity towards the
MMR deficient cell line at similar concentration ranges.
-80
-60
-40
-20
0
20
40
60
80
200
250
300
350
400
e
l
l
i
p
t
i
c
i
t
y
(
m
i
l
l
i
d
e
g
r
e
e
s
)
w
ave
l
e
n
gth
(n
m)
Λ
e
na
nt
i
om
e
r
Δ
e
na
nt
i
om
e
r
-80
-60
-40
-20
0
20
40
60
80
200
250
300
350
400
e
l
l
i
p
t
i
c
i
t
y
(
m
i
l
l
i
d
e
g
r
e
e
s
)
w
ave
l
e
n
gth
(n
m)
0
5
10
15
20
25
30
0
5
10
15
20
25
30
35
A
b
s
o
r
b
a
n
c
e
a
t
4
4
0
n
m
(
m
A
U
)
T
i
me
(mi
n
)
A
B
C
-80
-60
-40
-20
0
20
40
60
80
200
250
300
350
400
e
l
l
i
p
t
i
c
i
t
y
(
m
i
l
l
i
d
e
g
r
e
e
s
)
w
ave
l
e
n
gth
(n
m)
Λ
e
na
nt
i
om
e
r
Δ
e
na
nt
i
om
e
r
0
0.2
0.4
0.6
0.8
1
1.2
0
0.2
0.4
0.6
0.8
1
V
i
a
b
l
e
F
r
a
c
t
i
o
n
Δ
-[R
h
(p
h
e
n
)(c
h
r
ys
i
)(P
P
O
)
2+
(μ
M
)
H
C
T1
16N
H
C
T1
16O
0
0.2
0.4
0.6
0.8
1
1.2
0
0.2
0.4
0.6
0.8
1
V
i
a
b
l
e
F
r
a
c
t
i
o
n
Λ
-[R
h
(p
h
e
n
)(c
h
r
ys
i
)(P
P
O
)
2+
(μ
M
)
H
C
T1
16N
H
C
T1
16O
S
10
Figure S3
:
(A)
A representative competition titration between [Rh(bpy)(chrysi)(PPO)]
2+
and [Rh(bpy)
2
(chrysi)]
3+
to a hairpin DNA sequence containing a single CC mismatch.
As
the concentration of [Rh(bpy)(chrysi)(PPO)]
2+
is increased, [Rh(bpy)
2
(chrysi)]
3+
is
competed out of the mismatched site and the amount of DNA photocleavage decreases
, as
seen in the gel image
.
The
photocleaved fraction of DNA is plotted against the
log([Rh(bpy)(chrysi)(PPO)
2+
,μM]) and fit with a
sigmoidal curve.
(B) Melting temperature
analysis of a 9
-
mer containing a central CC mismatch in the presence of
[Rh(bpy)
2
(chrysi)]Cl
3
(RhBBC) or [Rh(ph
en)(chrysi)(PPO)]Cl
2
(RhPPO) showed
comparable stabilization of the duplex with melting temperatures of
44.9 ± 0.6 and 41.3 ±
0.5 °C
, respectively.
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
-1
-0.5
0
0.5
1
1.5
2
2.5
f
r
a
c
t
i
o
n
p
h
o
t
o
c
l
e
a
v
e
d
l
og([R
h
(b
p
y)(c
h
r
ys
i
)(P
P
O
)
2+
,
μ
M
])
0
0
.
0
5
0
.
1
0
.
1
5
0
.
2
0
.
2
5
1
0
2
0
3
0
4
0
5
0
A
b
s
o
r
b
a
n
c
e
I
n
c
r
e
a
s
e
T
e
m
p
e
r
a
t
u
r
e
(
D
e
g
r
e
e
s
C
)
W
i
t
h
R
h
B
B
C
W
i
t
h
R
h
P
P
O
5’- C G
G
A
C
T
C C G
-3’
3’- G
C C
T
C
A
G
G
C -5’
A
B
S
11
Figure S4
:
A pH titration of the metalloinsertor [Rh(bpy)(chrysi)(PPO)]
2+
. Absorption
spectra as the
pH chan
g
es
from 6.52 to 9.98 are shown, and the inset show the absorbance
at 435nm vs pH and fit to a sigmoidal curve.
Figure S5
:
A pH titration of the metalloinsertor [Rh(
HDPA
)(chrysi)(PPO)]
2+
. Absorption
spectra as the pH chan
g
es
from 7.88 to 10.53
are shown, and the
inset show the
absorbance at 440
nm vs pH and fit to a sigmoidal curve.
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
6.52
7.45
8.24
8.5
8.9
9.98
0.10
0.12
0.14
0.16
0.18
0.20
0.22
4.5
5.5
6.5
7.5
8.5
9.5
10.5
A
b
s
o
r
b
a
n
c
e
a
t
4
3
5
n
m
pH
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
7.88
8.82
9.15
9.35
9.77
10.53
0.14
0.15
0.16
0.17
0.18
0.19
0.20
0.21
4.5
5.5
6.5
7.5
8.5
9.5
10.5
A
b
s
o
r
b
a
n
c
e
a
t
4
4
0
n
m
pH
S
12
Fi
gure S6
:
A pH titration of the metalloinsertor [Rh(
4,7
-
DMP)(chrysi)(PPO)]
2+
. Absorption
spectra as the pH chan
g
es from 5.60 to 9.98 are shown, and the inset show the absorbance
at 440nm vs pH and fit to a sigmoidal curve.
Figure S7
:
A pH titration of the metalloinsertor [Rh(
5,6
-
DMP)(chrysi)(PPO)]
2+
. Absorption
spectra as the pH chan
g
es from 6.24 to 9.78 are sh
own, and the inset show the absorbance
at 438nm vs pH and fit to a sigmoidal curve.
0
0.5
1
1.5
2
2.5
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
5.6
8.12
8.85
9.20
9.45
9.98
0.14
0.16
0.18
0.20
0.22
0.24
0.26
4.5
5.5
6.5
7.5
8.5
9.5
10.5
A
b
s
o
r
b
a
n
c
e
a
t
4
4
0
n
m
pH
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
6.24
7.99
8.72
8.97
9.28
9.78
0.08
0.10
0.12
0.14
0.16
0.18
4.5
5.5
6.5
7.5
8.5
9.5
10.5
A
b
s
o
r
b
a
n
c
e
a
t
4
3
8
n
m
pH
S
13
Figure S8
:
A pH titration of the metalloinsertor [Rh(DIP)(chrysi)(PPO)]
2+
. Absorption
spectra as the pH chan
g
es from 6.04 to 10.25 are shown, and the inset show the
abs
orbance at 436nm vs pH and fit to a sigmoidal curve.
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
6.04
7.67
8.33
8.75
9.08
10.25
0.08
0.09
0.09
0.10
0.10
0.1
1
0.1
1
0.12
0.12
0.13
4.5
5.5
6.5
7.5
8.5
9.5
10.5
A
b
s
o
r
b
a
n
c
e
a
t
4
3
6
n
m
pH
S
14
Figure S9
:
Absorption spectrum of [Rh(phen)(chrysi)(PPO)]
2+
in 1
-
octanol before and after
equilibration with aqueous solution (pH 7.4)
with replicates taken at
three
different
concentrations
. Each replicate set (before and after trace) is shown as a different color
(purple, orange, or blue) with the before trace shown in a darker shade and the after trace
shown in the lighter shade.
Figure S10
:
Absorption spectrum of [Rh(bpy)(chrysi)(PPO)
]
2+
in 1
-
octanol before and
after equilibration with aqueous solution (pH 7.4) with replicates taken at three different
concentrations. Each replicate set (before and after trace) is shown as a different color
(purple, orange, or blue) with the before trac
e shown in a darker shade and the after trace
shown in the lighter shade.
0
0.2
0.4
0.6
0.8
1
1.2
1.4
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
0
0.2
0.4
0.6
0.8
1
1.2
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
S
15
Figure S11
:
Absorption spectrum of [Rh(HDPA)(chrysi)(PPO)]
2+
in 1
-
octanol before and
after equilibration with aqueous solution (pH 7.4) with replicates taken at three different
concentrations. Each replicate set (before and after trace) is shown as a different color
(purple, orange, or blue) with the before trace s
hown in a darker shade and the after trace
shown in the lighter shade.
Figure S12
:
Absorption spectrum of [Rh(4,7
-
DMP)(chrysi)(PPO)]
2+
in 1
-
octanol before and
after equilibration with aqueous solution (pH 7.4) with replicates taken at three different
concentrations. Each replicate set (before and after trace) is shown as a different color
(purple, orange, or blue) with the before trace s
hown in a darker shade and the after trace
shown in the lighter shade.
0
0.2
0.4
0.6
0.8
1
1.2
1.4
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)
0
0.2
0.4
0.6
0.8
1
1.2
1.4
200
300
400
500
600
700
800
A
b
s
o
r
b
a
n
c
e
w
ave
l
e
n
gth
(n
m)