New photoactivatable analogues of glutathione disulfide

Article abstract of DOI:10.1055/s-2005-918518

New photoactivatable analogues of glutathione disulfide (GSSG) bearing new benzophenone-like photophores were synthesized by using an improved coupling reaction. Georg Thieme Verlag Stuttgart.

Full text of DOI:10.1055/s-2005-918518

PAPER
509
New Photoactivatable Analogues of Glutathione Disulfide
New
P
hotoactivata
a
ble Analog
n
ues of
G
lutathione
B
D
isulfide ernardi, Amadou Dicko, Gilbert Kirsch*
Laboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Université Paul Verlaine-Metz, 1 Boulevard Arago, 57078 METZ
Cedex 3, France
Fax +33(38)7315801; E-mail: kirsch@sciences.univ-metz.fr
Received 12 July 2005; revised 17 August 2005
O
CO2H
NH
H
Abstract: New photoactivatable analogues of glutathione disulfide
(GSSG) bearing new benzophenone-like photophores were synthe-
sized by using an improved coupling reaction.
N
HO2C
O
N
H
N3
O
S
S
O
Key words: glutathione disulfide, photoaffinity labeling, benzo-
phenone, heterocycles
O
N3
H
HN
N
CO2H
N
H
CO2H
O
Photoaffinity labeling (PAL) is a frequently used tech-
nique for studying the interactions of biologically active
Figure 1 The structure of N,N¢-bis(4-azidobenzoyl)glutathione di-
molecules. A photolabile analogue of the compound un- sulfide.
der investigation, which is stable in the dark, is converted
by illumination into a reactive intermediate. This, by
In this paper we report the synthesis of new photoactivat-
forming a covalent bond, stabilizes the interaction based
on biological affinity.
able probes (Figure 2) using GSSG as starting material.
We have chosen to synthesize analogues of GSSG bearing
a benzophenone-like moiety as photoactivatable group.
Indeed the benzophenone photophore8 has been described
as being among the most efficient group for labeling re-
ceptors and enzymes. Moreover, attention is increasingly
focused on heterocyclic analogues (thiophene, pyrrole) of
the benzophenone moiety.9
Many authors are interested in the study of apoptosis. The
apoptotic process is a genetically controlled programmed
cell death and is often deficient in cancer cells. In the lit-
erature there is a great number of reports on pro- or anti-
apoptotic enzymes, which have glutathione in its reduced
form (GSH) as substrate.1 Most of these studies2 are led
using the PAL technique. Indeed it is known that GSH
plays an important role in apoptosis.3 Recently, Filomeni
et al.4 showed that the oxidized form of GSH (GSSG) in-
duces also apoptosis. In mammalian cells, less than about
We report here the synthesis of three new photophores
and their use in the preparation of several photoactivatable
probes.
0.2% of the total GSH occurs as GSSG. Oxidation of GSH The new photophores were prepared by coupling the mo-
to form GSSG is carried out either by direct interaction noester of terephthalic acyl chloride on the thiophene or
with free radicals, or more often, when GSH acts as a co- pyrrole ring using a Friedel–Crafts reaction in the pres-
factor for antioxidant enzymes such as GSH peroxidases ence of AlCl3 as catalyst (Scheme 1). Esters 1a–c were
during the reduction of H2O2. GSSG is also formed when obtained in poor to good yields (Table 1) after chromato-
glutaredoxin reductase uses GSH to reduce the disulfide graphic purification. They were hydrolyzed under basic
bond in glutaredoxin.
medium to give the corresponding acids 2a–c in moderate
to good yields (Table 1). Those compounds were activat-
ed before their coupling to GSSG, as we will discuss later.
To the best of our knowledge, the only reference reporting
a GSSG photoactivatable analogue was reported in 1991
by D’Silva et al.5 They have proposed the synthesis of
N,N¢-bis-(4-azidobenzoyl)glutathione disulfide (Figure 1)
in 35% yield by coupling p-azidobenzoyl chloride on
GSSG in the presence of 4-(N,N-dimethylamino)pyridine
(DMAP) in a mixture aqueous NaOH–dioxane. This bi-
functional aryl azide derivative of GSSG was used to
study yeast glutathione reductase.6
O
O
OH
OR
i
X
iii
X
X
ii
O
O
2a–c
1a–c
We decided to investigate the role of GSSG in apoptosis.
Indeed, cancer cells study7 is the topic of intense current
interest in our laboratory.
Scheme 1 Reagents and conditions: (i) monoester terephthalic acid,
SOCl2, reflux, 2 h; (ii) AlCl3, CH2Cl2, r.t.; (iii) KOH (5 equiv), appro-
priate alcohol, r.t., overnight.
Two other compounds were also used to form GSSG
probes. Acid 2d is commercially available. Acid 2e was
prepared according to the literature procedure10 in a three-
SYNTHESIS 2006, No. 3, pp 0509–0513
Advanced online publication: 21.12.2005
DOI: 10.1055/s-2005-918518; Art ID: Z13505SS
© Georg Thieme Verlag Stuttgart · New York
x
x
.
x
x
.2
0
0
5

510
D. Bernardi et al.
PAPER
O
CO2H
NH
H
N
HO2C
O
N
H
O
O
Y
X
S
S
O
O
X
Y
O
H
HN
N
CO2H
N
H
CO2H
O
X = S, Y = CH=CH (a)
X = NH, Y = CH=CH (b)
X = N-Me, Y = CH=CH (c)
X = Y = CH=CH (d)
X = CH=CH, Y = S (e)
Figure 2 The structures of new photoactivatable analogues of GSSG.
in the presence of 6 equivalents of Et3N for 36 hours. Pu-
rification on silica gel yielded the expected analogues 4a–
e in good yields. These compounds were fully character-
ized by NMR and mass spectra. It may be noticed that
some signals of the GSSG backbone are missing in the 13C
NMR spectra (see experimental section). However, these
results are similar to those reported by Gartner et al.12
Table 1 Preparation of Esters 1 and Acids 2
Entry
X
Esters 1 (%)
Acids 2 (%)
a
b
c
S
94
17
23
90
88
91
NH
NMe
The ultraviolet absorption spectra of 2a–e and 4a–e were
measured in 0.05 M NaOH. In comparison with the refer-
ence acid 2d, we observed a bathochromic shift for all the
other acids. (Table 2) Acid 2b had the highest absorption
maximum at 316 nm. Concerning the probes 4a–e
(Table 1), 4b and 4e had absorption wavelengths higher
than 300 nm, i.e. at 319 nm and 307 nm, respectively. It
may be noticed that 4c showed another absorption maxi-
mum at 316 nm but with a lower intensity (e = 16810).
These different absorption maxima are very interesting
because the photoaffinity labeling can be performed at
wavelengths, which are non-destructive for biological
systems.
step synthesis from 2-bromothiophene. It should be noted
that 2e has not yet been used for its possible photophoric
properties.
The low yield reported by D’Silva et al.5 prompted us to
investigate other methods and more effective reagents for
the N-acylation of GSSG. Among the activating groups
usually employed in peptide chemistry,11 the N-oxysuc-
cinimidyl group was well suited to our purpose.
O
O
X
N
i
In conclusion, the synthesis of new photoactivatable ana-
logues of GSSG as well as the synthesis of new chro-
mophores have been accomplished. As expected, three of
the new photoactivatable analogues of GSSG exhibit
promising photochemical properties. This makes them
good photolabeling reagents for GSSG utilizing systems.
Moreover, the coupling reaction, which is simple and ef-
OH
Y
Y
O
X
O
O
O
O
2a–e
3a, 83%
3b, 85%
3c, 80%
3d, 88%
3e, 90%
X = S, Y = CH=CH (a)
X = NH, Y = CH=CH (b)
X = N-Me, Y = CH=CH (c)
X = Y = CH=CH (d)
X = CH=CH, Y = S (e)
Table 2 Ultraviolet Absorption of Acids 2 and GSSG Analogues 4
Scheme 2 Reagents and conditions: (i) NHS, DCC, THF, 0 °C,
overnight.
Entry
Acids 2
lmax
ea
Probes 4
lmax
ea
Acids 2a–e were activated by action of N-hydroxysuccin-
imide (NHS) in the presence of dicyclohexylcarbodiimide
(DCC) as shown in Scheme 2. As expected, the targeted
N-oxysuccinimidyl esters 3a–e were obtained in good
yields after filtration of the dicyclohexylurea (DCU)
formed during the reaction. It should be noted that 3a–e
were always contaminated with DCU. This impurity was
eliminated during the purification of the coupling step.
a
b
c
300
316
315
263
304
15330
20100
18290
45620
27630
268
319
259
264
307
19680
21810
22830
34580
29220
d
e
a In L mol–1 cm–1.
In the last step (Scheme 3), the coupling reactions with
GSSG were carried out in a mixture of MeCN–H2O (3:1)
Synthesis 2006, No. 3, 509–513 © Thieme Stuttgart · New York

PAPER
New Photoactivatable Analogues of Glutathione Disulfide
511
O
X
NH2
O
S
Y
H
HO
N
OH
i
N
H
O
NH
O
S
O
O
O
H
HO
N
OH
N
H
2
O
O
O
X = S, Y = CH=CH (a)
X = N, Y = CH=CH (b)
X = N-Me, Y = CH=CH (c)
X = Y = CH=CH (d)
4a, 80%
4b, 80%
4c, 77%
4d, 85%
4e, 88%
2
X = CH=CH, Y = S (e)
Scheme 3 Reagents and conditions: (i) 3a–e, Et3N (6 equiv), H2O, MeCN, r.t., 36 h
Ethyl 4-(1H-Pyrrole-2-carbonyl)benzoate (1b)
Mp 79–82 °C.
ficient, proceeds in partially aqueous solution under mild
conditions and is expected to be quite general for the syn-
thesis of a wide range of GSSG photoactivatable ana- 1H NMR (250 MHz, CDCl3): d = 9.76 (br s, 1 H), 8.14–8.17 (d, J =
8 Hz, 2 H), 7.92–7.95 (d, J = 8 Hz, 2 H), 7.19 (s, 1 H), 6.87 (s, 1 H),
6.37 (s, 1 H), 4.38–4.47 (q, J = 5.4 Hz, 2 H), 1.40–1.45 (t, J = 4.75
logues.
Hz, 3 H).
4-Benzoylbenzoic acid was purchased from Acros. Melting points
13C NMR (62.5 MHz, CDCl3): d = 183.95, 165.93, 141.93, 133.18,
were determined on a Stuart SMP3 melting point apparatus and are
130.95, 130.06, 129.51, 119.86, 111.36, 61.35, 14.29.
uncorrected. 1H and 13C spectra were recorded on a AC Bruker 250
GC–MS: m/z (%) = 243 (52), 214 (10), 198 (29), 170 (61), 149 (4),
94 (100), 66 (21), 45 (1).
MHz spectrometer in CDCl3 or DMSO-d6. Mass spectra measure-
ments were performed on a Hewlett-Packard 5971 A GC-MS spec-
trometer. MALDI spectra were recorded on a system equipped with
a pulsed nitrogen laser (337 nm), operating in positive-ion reflector
mode, using 19 kV acceleration voltage and a matrix of a-cyano-4-
hydroxycinnamic acid.
Ethyl 4-(1-Methyl-1H-pyrrole-2-carbonyl)benzoate (1c)
Mp 67–69 °C.
1H NMR (250 MHz, CDCl3): d = 8.11–8.14 (d, J = 8.5 Hz, 2 H),
7.81–7.84 (d, J = 8.5 Hz, 2 H), 6.95 (s, 1 H), 6.69–6.71 (d, J = 5.75
Hz, 1 H), 6.15–6.18 (d, J = 6.75 Hz, 1 H), 4.37–4.46 (q, J = 5.38 Hz,
2 H), 4.05 (s, 3 H), 1.39–1.45 (t, J = 4.83 Hz, 3 H).
Esters 1; General Procedure13
The appropriate monoalkylterephthalate was refluxed in SOCl2
with two drops of DMF until a homogeneous solution was obtained.
The excess SOCl2 was then removed by evaporation under reduced
pressure leaving crude acyl chloride, which was not further purified.
13C NMR (62.5 MHz, CDCl3): d = 185.09, 165.31, 143.39, 132.52,
130.06, 129.09, 128.71, 123.15, 108.27, 61.11, 37.29, 14.13.
GC–MS: m/z (%) = 257 (37), 228 (49), 212 (37), 184 (100), 108
(47).
A solution of appropriate acyl chloride (10 mmol) in anhyd CH2Cl2
(15 mL) was added dropwise at r.t. over 10 min to a stirred mixture
of the appropriate pyrrole (10 mmol) and anhyd AlCl3 (1.60 g, 12
mmol) in the same solvent. After being stirred at r.t. for 30 min, the
reaction mixture was quenched with ice and sat. aq NaHCO3 (15
mL). The organic phase was separated and combined with the
CHCl3 extracts (2 × 15 mL) of the aqueous phase. The combined or-
ganic phases were washed with H2O, dried over MgSO4, and evap-
orated under reduced pressure. The crude residue was
chromatographed on silica gel column, with EtOAc–cyclohexane
(5:95) as eluent.
Acids 2; General Procedure14
Esters 1 (5 mmol) were dissolved in the appropriate anhyd alcohol
(20 mL). KOH (1.4 g, 25 mmol) was added to the solution and the
mixture was stirred at r.t. overnight. The solvent was removed under
reduced pressure and the residue was dissolved in 1 M HCl (30 mL).
The solution was then extracted with CH2Cl2 (2 × 20 mL). The com-
bined organic phase was washed with H2O (2 × 20 mL), dried over
MgSO4 and removed under reduced pressure.
In the case of thiophene, the acyl chloride was mixed with anhyd
AlCl3 in anhyd CH2Cl2 and thiophene (0.84 g, 10 mmol) was slowly
added. After being stirred at r.t. for 1.5 h, the reaction mixture was
quenched with ice and 1 M tartaric acid (15 mL).
4-(2-Thienylcarbonyl)benzoic Acid (2a)
Mp 200 °C.
IR (neat): 3100–2359 (br, OH), 1700 (C=O), 1627 (C=O), 1413,
1278 cm–1.
1H NMR (250 MHz, CDCl3 + DMSO-d6): d = 8.23–8.26 (d, J = 8.25
Hz, 2 H), 7.92–7.96 (d, J = 8.25 Hz, 2 H), 7.77–7.79 (d, J = 5.75 Hz,
1 H), 7.64 (d, J = 4.75 Hz, 1 H), 7.19 (t, J = 4 Hz, 1 H).
13C NMR (62.5 MHz, CDCl3 + DMSO-d6): d = 185.05, 164.91,
140.80, 139.24, 133.95, 133.78, 132.22, 127.67, 127.51, 126.93.
Methyl 4-(2-Thienylcarbonyl)benzoate (1a)
Mp 143–146 °C.
IR (neat): 2361, 1717 (C=O), 1623 (C=O), 1409, 1280 cm–1.
1H NMR (250 MHz, CDCl3): d = 8.14–8.18 (d, J = 8Hz, 2 H), 7.88–
7.91 (d, J = 8 Hz, 2 H), 7.65–7.77 (d, J = 6.75 Hz, 1 H), 7.62–7.63
(d, J = 4.75 Hz, 1 H), 7.16–7.19 (t, J = 4.3 Hz, 1 H), 3.96 (s, 3 H).
13C NMR (62.5 MHz, CDCl3): d = 186.45, 166.22, 143.17, 141.79,
135.15, 133.14, 129.65, 129.07, 128.13, 52.42.
UV–Vis: lmax (e) = 271 (14500), 300 (15330) nm.
4-(1H-Pyrrole-2-carbonyl)benzoic Acid (2b)
Mp 190–192 °C.
GC–MS: m/z (%) = 246 (28), 231 (1), 215 (11), 187 (10), 163 (12),
111 (100), 83 (7).
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512
D. Bernardi et al.
PAPER
1H NMR (250 MHz, CDCl3): d = 10.64 (br s, 1 H), 8.08–8.11 (d,
J = 7.75 Hz, 2 H), 7.83–7.86 (d, J = 7.75 Hz, 2 H), 7.12 (s, 1 H),
6.79 (s, 1 H), 6.26 (s, 1 H).
13C NMR (62.5 MHz, CDCl3): d = 195.46, 168.96, 161.18, 142.98,
136.48, 133.19, 130.45, 130.07, 129.92, 128.53, 128.01, 25.63.
N-Hydroxysuccinimidyl-5-benzoylthiophene-2-carboxylate
(3e)
Mp 137–139 °C.
1H NMR (CDCl3): d = 8.01–8.02 (d, J = 4.25 Hz, 1 H), 7.87–7.90
(d, J = 7.5 Hz, 2 H), 7.63–7.68 (m, 2 H), 7.51–7.57 (m, 2 H), 2.92
(s, 4 H).
13C NMR (62.5 MHz, CDCl3): d = 168.73, 136.81, 136.01, 133.53,
133.25, 129.34, 128.72.
13C NMR (62.5 MHz, CDCl3): d = 183.94, 167.79, 141.99, 133.33,
130.78, 129.53, 129.53, 128.48, 119.91, 110.78.
UV–Vis: lmax (e) = 255.5 (11730), 316.5 (20100) nm.
4-(1-Methyl-1H-pyrrole-2-carbonyl)benzoic Acid (2c)
Mp 193–195 °C.
1H NMR (250 MHz, CDCl3 + DMSO-d6): d = 8.18–8.21 (d, J = 8.25
Hz, 2 H), 7.85–7.88 (d, J = 8.25 Hz, 2 H), 6.97 (s, 1 H), 6.72 (s, 1
H), 6.,19 (s, 1 H), 4.06 (s, 3 H).
13C NMR (62.5 MHz, CDCl3 + DMSO-d6): d = 184.44, 166.97,
142.76, 133.93, 129.39, 128.83, 128.69, 128.05, 122.58, 107.78,
36.67.
GSSG Analogues 4; General Procedure
N-Hydroxysuccinidimyl esters 3 (1 equiv) were added at r.t. to a so-
lution of GSSG (10 mg, 0.016 mmol) in a solution of MeCN (0.3
mL) and H2O (0.1 mL) in the presence of Et3N (0.013 mL, 6 equiv).
The reaction mixture was then stirred at r.t. for 36 h and concentrat-
ed under reduced pressure. The crude residue was chromatographed
on silica gel column, with MeCN–H2O (9:1) as eluent.
UV–Vis: lmax (e) = 255.5 (11220), 315 (18290) nm.
N-Hydroxysuccinidimyl Esters 3, General Procedure15
A solution of acid 2 (1 mmol) and N-hydroxysuccinimide (115 mg,
1 mmol) in anhyd THF (2 mL) was cooled to 0 °C. DCC (216 mg,
1.05 mmol) was added and stirring was continued at this tempera-
ture for 4 h. The reaction mixture was allowed to stand overnight in
a refrigerator and then filtered. The solution was concentrated under
reduced pressure. The residue was triturated in Et2O–light petro-
leum (1:1), filtered out, and then dried in vacuo to afford the expect-
ed compound.
N,N¢-Bis[4-(2-thienylcarbonyl)benzoyl]glutathione Disulfide
(4a)
Yield: 80%; mp 230 °C (decomp.).
1H NMR (250 MHz, D2O): d = 7.92–7.95 (m, 4 H), 7.78–7.82 (m,
10 H), 7.63–7.66 (m, 4 H), 4.77 (t, 2 H, Cysa), 4.49 (t, 2 H, Glua),
3.77 (s, 4 H, Glya), 3.28 (dd, 2 H, Cysb1), 2.96 (dd, 2 H, Cysb2),
2.58 (m, 4 H, Glug), 2.40 (m, 2 H, Glub1), 2.20 (m, 2 H, Glub2).
13C NMR (62.5 MHz, D2O): d = 190.24, 175.57, 171.93, 162.25,
145.51, 145.06, 136.37, 136.12, 133.52, 130.28, 129.54, 128.69,
52.53, 38.70, 31.83, 27.09 (some peaks were not observed).
Compounds 3a–e were always contaminated with DCU.
N-Hydroxysuccinimidyl-4-(2-thienylcarbonyl)benzoate (3a)
Mp 193–195 °C.
1H NMR (250 MHz, CDCl3): d = 8.25–8.29 (d, J = 8.5 Hz, 2 H),
7.97–7.93 (d, J = 8.5 Hz, 2 H), 7.79–7.81 (d, J = 6 Hz, 1 H), 7.61–
7.62 (d, J = 4.25 Hz, 1 H), 7.18–7.22 (t, J = 4.63 Hz, 1 H), 2.94 (s,
4 H).
MS (MALDI–TOF): m/z = 1063 [M + Na]+.
UV–Vis: lmax (e) = 268 (19680), 292 (16790) nm.
N,N¢-Bis[4-(1H-pyrrole-2-carbonyl)benzoyl]glutathione Disul-
fide (4b)
Yield: 80%; mp 239 °C (decomp.).
13C NMR (62.5 MHz, CDCl3): d = 186.99, 168.96, 162.03, 143.58,
142.89, 135.38, 135.35, 130.65, 129.21, 128.31, 128.04, 25.61.
1H NMR (250 MHz, D2O): d = 7.79 (m, 8 H), 7.39 (m, 2 H), 6.90
(m, 2 H), 6.42 (m, 2 H), 4.75 (t, 2 H, Cysa), 4.42 (t, 2 H, Glua), 3.79
(s, 4 H, Glya), 3.35 (dd, 2 H, Cysb1), 2.96 (dd, 2 H, Cysb2), 2.54 (m,
4 H, Glug), 2.36 (m, 2 H, Glub1), 2.15 (m, 2 H, Glub2).
13C NMR (62.5 MHz, D2O): d = 185.72, 175.67, 171.83, 168.99,
140.50, 136.49, 130.16, 129.07, 128.65, 127.29, 122.59, 111.47,
52.47, 38.44, 31.87, 26.97 (some peaks were not observed).
N-Hydroxysuccinimidyl-4-(1H-pyrrole-2-carbonyl)benzoate
(3b)
Mp 162–165 °C.
1H NMR (250 MHz, CDCl3): d = 9.78 (br, 1 H), 8.24–8.27 (d, J =
8.25 Hz, 2 H), 7.97–8.00 (d, J = 8.25 Hz, 2 H), 7.21 (s, 1 H), 6.85
(s, 1 H), 6.38 (s, 1 H), 2.94 (s, 4 H).
13C NMR (62.5 MHz, CDCl3): d = 183.33, 169.11, 161.30, 143.76,
130.54, 129.10, 127.63, 126.47, 120.35, 111.56.
MS (MALDI–TOF): m/z = 1007 [M + H]+, 1029 [M + Na]+.
UV–Vis: lmax (e) = 255.5 (18800), 319.5 (21810) nm.
N-Hydroxysuccinimidyl-4-(1-methyl-1H-pyrrole-2-carbon-
yl)benzoate (3c)
N,N¢-Bis-[4-(1-methyl-1H-pyrrole-2-carbonyl)benzoyl]gluta-
thione Disulfide (4c)
Mp 202–204 °C.
Yield: 77%; mp 220 °C (decomp.).
1H NMR (250 MHz, CDCl3): d = 8.21–8.25 (d, J = 8.5 Hz, 2 H),
7.87–7.91 (d, J = 8.5 Hz, 2 H), 6.98 (s, 1 H), 6.67–6.68 (d, J = 3.75
Hz, 1 H), 6.17–6.20 (d, J = 6.75 Hz, 1 H), 4.07 (s, 3 H), 2.94 (s, 4 H).
13C NMR (62.5 MHz, CDCl3): d = 184.55, 168.08, 160.84, 145.54,
132.49, 130.83, 130.36, 129.81, 127.16, 123.61, 108.72, 37.53,
25.69.
1H NMR (250 MHz, D2O): d = 7.86–7.89 (d, 4 H), 7.73–7.76 (d, 4
H), 7.24 (s, 2 H), 6.80 (s, 2 H), 6.25 (s, 2 H), 4.75 (t, 2 H, Cysa),
4.42 (t, 2 H, Glua), 3.97 (s, 6 H), 3.71 (s, 4 H, Glya), 3.23 (dd, 2 H,
Cysb1), 2.96 (dd, 2 H, Cysb2), 2.54 (m, 4 H, Glug), 2.36 (m, 2 H,
Glub1), 2.15 (m, 2 H, Glub2).
13C NMR (62.5 MHz, D2O): d = 186.54, 175.67, 171.86, 168.83,
141.85, 136.15, 134.72, 129.63, 129.19, 127.10, 125.69, 109.02,
52.49, 46.69, 38.45, 36.86, 31.84, 27.09 (some peaks were not ob-
served).
N-Hydroxysuccinimidyl-4-benzoylbenzoate (3d)
Mp 195–198 °C.
1H NMR (CDCl3): d = 8.24–8.28 (d, J = 8 Hz, 2 H), 7.88–7.91 (d,
J = 8 Hz, 2 H), 7.78–7.82 (d, J = 6.75 Hz, 2 H), 7.61–7.67 (m, 1 H),
7.49–7.54 (m, 2 H), 2.94 (s, 4 H).
MS (MALDI–TOF): m/z = 1037 [M + Na]+.
UV–Vis: lmax (e) = 259 (22830), 316 (16810) nm.
Synthesis 2006, No. 3, 509–513 © Thieme Stuttgart · New York

PAPER
New Photoactivatable Analogues of Glutathione Disulfide
(2) (a) Quian, Y.; Grant, C. E.; Westlake, C. J.; Zhang, D.;
513
N,N¢-Bis-(4-benzoylbenzoyl)glutathione Disulfide (4d)
Yield: 85%; mp 240 °C (decomp.).
Lander, P. A.; Shepard, R. L.; Dantzig, A. H.; Cole, S. P. C.;
Deeley, R. G. J. Biol. Chem. 2002, 277, 35225. (b) Furuta,
K.; Tomokiyo, K.; Kuo, M. T.; Ishikawa, T.; Suzuki, M.
Tetrahedron 1999, 55, 7529. (c) Furuta, K.; Hosoya, T.;
Tomokiyo, K.; Okuda, S.; Kuniyasu, A.; Nakayama, H.;
Ishikawa, T.; Suzuki, M. Bioorg. Med. Chem. Lett. 1999, 9,
2661.
1H NMR (250 MHz, D2O): d = 7.92–7.95 (m, 4 H), 7.78–7.82 (m,
10 H), 7.63–7.66 (m, 4 H), 4.77 (t, 2 H, Cysa), 4.49 (t, 2 H, Glua),
3.77 (s, 4 H, Glya), 3.28 (dd, 2 H, Cysb1), 2.96 (dd, 2 H, Cysb2),
2.58 (m, 4 H, Glug), 2.40 (m, 2 H, Glub1), 2.20 (m, 2 H, Glub2).
13C NMR (62.5 MHz, D2O): d = 199.58, 177.,92, 175.72, 171.82,
168.85, 139.47, 137.42, 136.25, 133.86, 130.40, 130.32, 128.68,
127.35, 55.30, 52.61, 43.39, 38.67, 31.96, 27.10.
(3) Hall, A. G. Eur. J. Clin. Invest. 1999, 29, 238.
(4) Filomeni, G.; Rotilio, G.; Ciriolo, M. R. FASEB J. 2003, 17,
64.
(5) D’Silva, C.; Seddon, A. P.; Douglas, K. T. J. Chem. Soc.,
Perkin Trans. 1 1981, 3029.
(6) Douglas, K. T.; Seddon, A. P.; D’Silva, C.; Bunni, M.
Biochem. Soc. Trans. 1982, 10, 124.
(7) Brault, L.; Migianu, E.; Néguesque, A.; Battaglia, E.;
Bagrel, D.; Kirsch, G. Eur. J. Med. Chem. 2005, in press.
(8) Dorman, G.; Prestwich, G. D. Biochemistry 1994, 33, 5661.
(9) (a) Bosca, F.; Miranda, M. A. J. Photochem. Photobiol., B:
Biology 1998, 43, 1. (b) Lahoz, A.; Hernandez, D.; Miranda,
M. A.; Perz-Prieto, J.; Morera, I. M.; Castell, J. V. Chem.
Res. Toxicol. 2001, 14, 1486. (c) Moser, J.; Hye, A.; Lovell,
W. W.; Earl, L. K.; Castell, J. V.; Miranda, M. A. Toxicol. in
Vitro 2001, 15, 333. (d) Arnold, D. R.; Birtwell, R. J. J. Am.
Chem. Soc. 1973, 95, 4599.
MS (MALDI–TOF): m/z = 1029 [M + H]+, 1051 [M + Na]+, 1067
[M + K]+.
UV–Vis: lmax (e) = 264.5 (34580) nm.
N,N¢-Bis-(5-benzoylthiophene-2-carbonyl)glutathioneDisulfide
(4e)
Yield: 88%; mp 245 °C (decomp.).
1H NMR (250 MHz, D2O): d = 7.71–7.81 (m, 8 H), 7.59 (m, 6 H),
4.77 (t, 2 H, Cysa), 4.42 (t, 2 H, Glua), 3.79 (s, 4 H, Glya), 3.35 (dd,
2 H, Cysb1), 2.96 (dd, 2 H, Cysb2), 2.55 (m, 4 H, Glug), 2.36 (m, 2
H, Glub1), 2.15 (m, 2 H, Glub2).
13C NMR (62.5 MHz, D2O): d = 190.35, 175.61, 171.89, 168.6,
141.61, 139.83, 137.94, 137.19, 137.01, 129.39, 129.01, 127.42,
52.49, 38.46, 31.83, 27.06 (some peaks were not observed).
(10) Holland, G. F. US Patent, 4282246, 1981; Chem. Abstr.
1981, 95, 203733a.
MS (MALDI–TOF): m/z = 1063 [M + Na]+, 1079 [M + K]+.
(11) Green, T. W.; Wuts, P. G. M. Protective Groups in Organic
UV–Vis: lmax (e) = 307 (29220) nm.
Synthesis, 2nd ed.; Wiley: New York, 1991.
(12) Gartner, C. A.; Wen, B.; Wan, J.; Becker, R. S.; Jones, G. II;
Gygi, S. P.; Nelson, S. D. Biochemistry 2005, 44, 1846.
(13) Cadamuro, S.; Degani, I.; Dughera, S.; Fochi, R.; Gatti, A.;
Piscopo, L. J. Chem. Soc., Perkin Trans. 1 1993, 273.
(14) Zu, J.; Wang, X.; Chen, Y.; Jiang, X.; Chen, X.; Li, Z. J.
Org. Chem. 2004, 69, 6221.
Acknowledgment
We are grateful to Dr. G. Sommen for his help in bibliographic re-
search and to Dr. S. Hesse for her helpful discussions. We are gra-
teful to LSMCL Metz (France) for mass spectra.
(15) Anderson, G. W.; Zimmerman, J. E.; Callahan, F. M. J. Am.
Chem. Soc. 1964, 86, 1839.
References
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Synthesis 2006, No. 3, 509–513 © Thieme Stuttgart · New York