Multicomponent assembly of proposed DNA precursors in water

Article abstract of DOI:10.1021/ja306176n

We propose a novel pathway for the prebiotic synthesis of 2′-deoxynucleotides. Consideration of the constitutional chemical relationships between glycolaldehyde and β-mercapto-acetaldehyde, and the corresponding proteinogenic amino acids, serine and cysteine, led us to explore the consequences of the corresponding sulfur substitution for our previously proposed pathways leading to the canonical ribonucleotides. We demonstrate that just as 2-aminooxazole-an important prebiotic ribonucleotide precursor-is readily formed from glycolaldehyde and cyanamide, so is 2-aminothiazole formed from β-mercapto-acetaldehyde and cyanamide in water at neutral pH. Indeed, both the oxazole and the thiazole can be formed together in a one-pot reaction, and can be co-purified by crystallization or sublimation. We then show that 2-aminothiazole can take part in a 3-component carbon-carbon bond-forming reaction in water that leads to the diastereoselective synthesis of masked 2′-thiosugars regiospecifically tethered to purine precursors, which would lead to 2′-deoxynucleotides upon desulfurization. The possibility of an abiotic route to the 2′-deoxynucleotides provides a new perspective on the evolutionary origins of DNA. We also show that 2-aminothiazole is able to sequester, through reversible aminal formation, the important nucleotide precursors glycolaldehyde and glyceraldehyde in a stable, crystalline form.

Full text of DOI:10.1021/ja306176n

Article
pubs.acs.org/JACS
Multicomponent Assembly of Proposed DNA Precursors in Water
,
†,§
‡
§
Matthew W. Powner,* Shao-Liang Zheng, and Jack W. Szostak
†Department of Chemistry, University College London, Christopher Ingold Laboratories, 20 Gordon Street, London, WC1H 0AJ,
U.K.
‡
Harvard X-ray Crystallography Centre, 12 Oxford Street, Cambridge, Massachusetts 02138, United States
§
Howard Hughes Medical Institute and Department of Molecular Biology and Center for Computational and Integrative Biology,
Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, United States
*
S Supporting Information
ABSTRACT: We propose a novel pathway for the prebiotic
synthesis of 2′-deoxynucleotides. Consideration of the
constitutional chemical relationships between glycolaldehyde
and β-mercapto-acetaldehyde, and the corresponding protei-
nogenic amino acids, serine and cysteine, led us to explore the
consequences of the corresponding sulfur substitution for our
previously proposed pathways leading to the canonical
ribonucleotides. We demonstrate that just as 2-aminooxazole−an important prebiotic ribonucleotide precursor−is readily
formed from glycolaldehyde and cyanamide, so is 2-aminothiazole formed from β-mercapto-acetaldehyde and cyanamide in
water at neutral pH. Indeed, both the oxazole and the thiazole can be formed together in a one-pot reaction, and can be co-
purified by crystallization or sublimation. We then show that 2-aminothiazole can take part in a 3-component carbon−carbon
bond-forming reaction in water that leads to the diastereoselective synthesis of masked 2′-thiosugars regiospecifically tethered to
purine precursors, which would lead to 2′-deoxynucleotides upon desulfurization. The possibility of an abiotic route to the 2′-
deoxynucleotides provides a new perspective on the evolutionary origins of DNA. We also show that 2-aminothiazole is able to
sequester, through reversible aminal formation, the important nucleotide precursors glycolaldehyde and glyceraldehyde in a
stable, crystalline form.
INTRODUCTION
Although RNA has often been considered as a candidate for
■
1
,2
the first biopolymer of life, extant biology utilizes two
chemically distinct, but related, nucleotide polymers, RNA and
DNA. DNA is usually viewed as a late evolutionary adaptation
A plausible abiotic chemical route to the canonical nucleotides
is a major goal in origins of life research.
demonstrated the first chemical steps toward a divergent
pyrimidine and purine ribonucleotide synthesis. A one-pot
1
−3
Recently, we
4
3
of earlier RNA-based life. However, it would not be possible to
make DNA without deoxyribonucleotides, and yet in the
absence of DNA there is no obvious reason for the evolution of
the biochemical pathways for the synthesis of deoxyribonucleo-
tides. Only after the emergence of DNA as an important
cellular component would there have been a strong selective
pressure favoring the emergence of biochemical pathways for
the synthesis of deoxyribonucleotides from ribonucleotides.
Although the advantages of DNA as a medium for information
storage are clear, there has not until now been any reasonable
hypothesis for how DNA could be ‘invented’ by primitive cells
in the absence of pre-existing deoxyribonucleotides. In this
paper, we propose a hypothesis for the prebiotic synthesis of 2′-
deoxyribonucleotides.
multicomponent reaction was demonstrated to stereoselectively
tether and consequently regiospecifically glycosylate purine
precursors and masked pentose sugars, while concurrently
furnishing known pyrimidine precursors (Scheme 1).
Scheme 1. Proposed Multicomponent Ribonucleotide
Syntheses
Biochemically, 2′-deoxyribonucleotides are synthesized by
5
chemo- and regio-specific reduction of ribonucleotides. In the
absence of the complex and energetically costly enzymatic
resources required to regiospecifically deoxygenate ribonucleo-
tides, abiological 2′-deoxygenation of nucleotides requires site-
Received: June 28, 2012
Published: July 27, 2012
©
2012 American Chemical Society
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Article
specific ribo- or arabino-nucleoside deoxygenation. In light of
the difficulty of regioselective nucleotide deoxygenation in the
absence of sophisticated enzymatic control, it would seem that
regiospecific positioning of a group/atom with the appropriate
latent potential reactivity could form the basis of an alternative
pathway to the 2′-deoxyribonucleotides. In principle, a
sufficiently chemoselective reactivity difference could be
exploited to generate both ribonucleotides and 2′-deoxyribo-
nucleotides, simultaneously.
reactivity of glycolaldehyde 3 and β-mercapto-acetaldehyde 6
with cyanamide 2 could lead, via 2-aminooxazole 1 and 2-
1
,3
aminothiazole 7, to ribonucleotides and 2′-deoxyribonucleo-
1
0,11
tides, by desulfurization of C2′ (Scheme 2).
The importance of 2-aminothiazole 7, and its derivatives, has
1
2
been recognized in medicinal chemistry, but though the
13
synthesis of thiazole 7 in water has been reported, the
potential relevance of thiazole 7 to the origins of life remains
unexplored. The formation of oxazole 1 from cyanamide 2 and
glycolaldehyde 3 at neutral pH requires stoichiometric (or
excess) cyanamide 2 and general acid−base catalysis. Aldehyde
, upon imine formation with cyanamide 2, has, like 3, the
1
RESULTS AND DISCUSSION
■
6
Constitutional analysis of DNA, with respect to RNA, in light
6
potential to undergo 5-exo-dig cyclization to furnish imine 14
or hemiaminal 15) as shown in Scheme 3. Furthermore, we
of the low dissociation energy of C−S bonds, suggests that
(
regiospecific positioning of sulfur at C2′ could result in the
chemical differentiation required for the divergent synthesis of
DNA and RNA monomers from common precursors (Scheme
expected the reaction to proceed more rapidly at neutral pH
and be less prone to stall at intermediates en route to thiazole 7
due to the increased nucleophilicity of sulfur and greater
aromaticity of thiazoles with respect to oxazoles.
2
).
To test this supposition, β-mercapto-acetaldehyde 6 (8.5
Scheme 2. Proposed Multicomponent Deoxyribonucleotide
Syntheses
mM) and 2 (15, 26, or 50 mM) were incubated in D O at pD 7
2
and 20 °C and a rapid (ca. 10 min) quantitative conversion of
aldehyde 6 to hemiaminal 15 was observed (see the Supporting
Information Figure S1). Over the course of several hours, 15
dehydrated to furnish thiazole 7 (see Supporting Information
14
Figures S2 and S3). It is of note that excess aldehyde 6 did
not lead to further reaction of thiazole 7. Moreover, at elevated
concentration (>150 mM) 2-aminothiazole 7 was observed to
directly crystallize from water, and a crystal structure of thiazole
Oxazole 1−a key ribonucleotide precursor−is derived from
15,16
7
is shown in Scheme 3.
Next we investigated the
1
,3,7
cyanamide 2 and glycolaldehyde 3.
The C2-carbon atom of
concomitant synthesis of oxazole 1 and thiazole 7. In the
presence of excess cyanamide 2 (2.2−5 equiv), rapid and
stoichiometric conversion of glycolaldehyde 3 (1 equiv) and β-
mercapto-acetaldehyde 6 (1 equiv) to hemiaminals 11 and 15,
respectively, was observed in water. At high pH/pD (pH/pD >
aldehyde 3 is formally delivered to a ribonucleotide as the C2′
sugar carbon. It is of note that aldehyde 3 is also the aldehyde
precursor of serine 4, a proteinogenic amino acid, via Strecker
8
synthesis with ammonium cyanide (Scheme 3). The constitu-
tional similarity of serine 4 and cysteine 5, and their aldehyde
precursors 3 and β-mercapto-acetaldehyde 6, suggests that both
9
6
) specific base catalysis or in 1 M phosphate solution (pH/pD
.5−7.5) general acid−base catalysis furnish oxazole 1 and
3
and 6 must be considered within our exploration of prebiotic
thiazole 7 over the course of 24−36 h at ambient temperature
9
azole synthesis. Specifically, we propose that the divergent
13
(
Scheme 3; see Supporting Information Figure S8−S10). At
neutral pD, selective dehydration of 15 to liberate thiazole 7
was observed. Limiting cyanamide 2 (2/3/6 ∼ 0.8:1:1) also
resulted in the specific generation of thiazole 7, even in 1 M
phosphate solution (pH 7; See Supporting Information Figure
S9). It is likely that the rapid 5-exo-dig cyclization of imine 12/
hemiaminal 13 leads to the selective sequestration of
cyanamide 2 in thiazole 7.
Scheme 3. Constitutional Analysis: Strecker-Type Synthesis
of Amino Acids (Red Box) and Azole Synthesis in Water
(Blue Box)
We did not observe significant homo- and hetero-aldol
reaction during the reaction of glycolaldehyde 3 and
mercaptoacetaldehyde 6 with either excess or limiting
cyanamide 2 between ambient temperature and 60 °C in 1
M phosphate solution at pH 7. Interestingly, we also found
that, like oxazole 1, thiazole 7 is easily sublimed (see
Supporting Information Figure S4), suggesting possible geo-
chemical routes to the distribution, separation, and purification
of thiazole 7 under abiotically plausible conditions. Sublimation
from a mixture of oxazole 1 and thiazole 7 resulted in the
isolation of both azoles as colorless crystalline solids. Partial
sublimation of a mixture of oxazole 1 and thiazole 7 (1:1)
resulted in the isolation 4:1−2:1 oxazole 1/thiazole 7, at 50 °C
between 1 and 24 h at ambient pressure, indicating the more
rapid but comparable sublimation rate of oxazole 1 relative to
thiazole 7. Complete sublimation was observed after 2 d at 60
°
C to afford ∼1:1 mixture of crystalline azoles at ambient
pressure.
1
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We then examined the multicomponent reaction of thiazole
, 4-amino-imidazole 5-carboxamide 16, and a series of
Scheme 5. Three-Component Reaction of 2-Aminothiazole
7, 4-Aminoimidazole-5-carboxamide 16, and Glyceraldehyde
22
17
7
aldehydes. Initially model aliphatic aldehydes were investigated
to avoid issues of annulation control. Equimolar quantities of
thiazole 7 and aliphatic aldehydes (including formaldehyde,
acetaldehyde, propionaldehyde, and cyclobutaldehyde) with
excess amino-imidazole 16 (1.2−3 equiv) furnished high yields
(
5
75−90%) of thiazolines 17, 18, 19, and 20, respectively, at pH
. The multicomponent reactions were found to be robust at
multi-gram scale and comparable diastereoselectivity was
observed in the reaction of aliphatic aldehydes with oxazole 1
or thiazole 7 and amino-imidazole 16 (via imine 21 R = alkyl;
3
d.r.= 2.5:1−4:1 threo/erythro). Compounds 17, threo-18 and
erythro-18, and the major isomers of 19 and 20 were isolated by
fractional crystallization, and the crystal structures are shown in
1
9
Scheme 4.
Scheme 4. Three-Component Reaction of 2-Aminothiazole
, 5-Amino-imidazole-4-carboxamide 16, and Various
7
Aliphatic Aldehydes
S35). Isolation of the two major isomers, crystallization, and X-
22
ray diffraction proved their xylo- and lyxo-configuration. The
diastereoselectivity obtained, upon carbon−carbon bond
formation is proposed to result from intramolecular hydro-
gen-bonding and concomitant steric impedance of thiazole
addition to the Si-face of imine 21 (see Supporting Information
3
,23
Scheme S1).
Although the facial selectivity of the
nucleophile 7 has been relaxed with respect to the comparable
reaction of 1, the facial selectivity of the glyceraldehyde-imine
3
2
1 (R = CH OH) is retained. This is important with respect
2
to purine nucleotide synthesis by 3′-purine tethering, which
necessarily requires N3′-displacement and C3′-inversion.
Furthermore, as well as regiospecific tethering of 16 and
consequent delivery of N1 to the anomeric carbon atom,
reaction of thiazole 7 with imine 21 regiospecifically delivers
sulfur to C2′, which is of significant interest with regards to the
potential synthesis of 2′-deoxynucleotides by C2′-desulfuriza-
tion.
To continue our investigation into the potential of
multicomponent assembly of nucleotides we studied the
participation of thiazole 7 in carbon−carbon bond-forming
reaction with α-hydroxyaldehydes and imines derived from α-
The multicomponent tricyclic thiazoline products 17−24
were uncontaminated with homoaldolization and homo-
Mannich byproducts. Additionally, oxazole 1 is known to
react efficiently with α-hydroxyaldehydes in a pH-dependent
20
hydroxyaldehydes in water. Incubation of equimolar aldehyde
, 2-aminothiazole 7, and amino-imidazole 16 for 5 d in water
3
at pH 4−6 at 20 °C gave a high yield of C -thiazoline 23 (80%
4
3
isolated yield) as 1.2:1 erythro/threo mixture of diastereomeric
products. Surprisingly, this signified that the facial selectivity of
thiazole 7 is significantly relaxed with respect to oxazole 1 and
bimolecular domino-reaction. In striking contrast, no signifi-
1
cant carbon−carbon bond formation was observed by H NMR
analysis upon incubation of thiazole 7 with either glycolalde-
imine 21 (R = CH OH). However, both erythro-23 and threo-
hyde 3 or rac-glyceraldehdye 22 in D O (pD 3−9, 4−100 °C).
2
2
2
3 were formed with complete annulation control (>99%, see
Instead, over a period of 4 days at pD 7, crystal formation was
1
H NMR spectrum shown in Supporting Information Figure
observed from an aqueous solution containing 3 (100 mM) and
24
S28; single crystal X-ray structures of erythro-23 and threo-23
thiazole 7 (100 mM) at 20 °C. Single crystal X-ray diffraction
22
are shown in Scheme 5).
of a crystal isolated directly from the reaction supernatant
25
Upon investigation of the parallel MCR with the C -synthon
proved that the structure was aminal 25 (Scheme 6).
3
glyceraldehyde 22, comparable efficiency in the synthesis of C5-
thiazoline 24 (70% isolated yield) was observed. Again,
exclusive 6-exo-trig annulation is observed; however, in this
case, 24 is furnished with significant diastereoselectivity (d.r.=
Though NMR studies confirmed that rac-22 formed
hemiaminal 26 and aminal 27 with thiazole 7 in D O, no
2
aminal crystallization was observed in solutions of rac-22 and 7,
even at significantly elevated concentrations (500 mM to 2M)
or with substoichiometric quantities of rac-22. To test whether
9:5:1.1:1, see Scheme 5 and Supporting Information Figure
1
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Scheme 6. Crystallization of Bis-(2-aminothiazole)-aminals
of Glycolaldehyde 3 and D-Glyceraldehyde 22 from Water at
pH 7
until now made it difficult to conceive of means for
accumulating large reservoirs of these essential starting
materials in a prebiotic context.
By exploring the multicomponent reactivity of thiazole 7,
together with aldehydes and the purine precursor amino-
imidazole 16, we have demonstrated a selective and high
yielding 3-component carbon−carbon bond-forming reaction
that chemospecifically furnishes masked-2′-thiosugars regiospe-
cifically tethered to an amino-imidazole purine-precursor. The
facial selectivity of thiazole 7, upon addition to α-
hydroxyaldehyde-imines, is quite relaxed (relative to oxazole
1
), resulting in low diastereoselectivity at C2′; however, this is
inconsequential with regards to the proposed deoxynucleotide
synthesis because C2′ would be rendered achiral following
desulfurization. In contrast, we observed very high diaster-
eoselectivity of nucleophilic addition of thiazole 7 to imine 21,
generating almost exclusively the lyxo and xylo isomers with
controlled N9−C1′ annulation. The conversion of the resulting
the change in solubility properties was due to the increased
number of hydroxyl moieties or perhaps due to the differential
solubility and packing properties of rac-27 with respect to the
achiral aminal 25, we investigated the solubility properties of
the aminal structures formed in a mixture of D-glyceraldehyde
C -thiazolines to 2′-deoxyribo-purine nucleotides requires three
5
additional steps: C3′-inversion to release the purine-precursor
from its tethered attachment to C3′, completion of the purine
2
2 and thiazole 7. Intriguingly, we found that at 100 mM
28
concentration, D-27 now crystallized from water at 20 °C.
NMR analysis showed a 1:2 ratio of 22/7 and recrystallization
heterocycle, and chemospecific desulfurization of C2′. These
steps are currently under investigation in our laboratories as
part of our continuing studies aimed at the abiotic synthesis of
(
10:1 H O/EtOH) allowed for crystallographic confirmation of
2
25
29
the aminal structure (Scheme 6).
2′-deoxyribonucleotides.
Finally, it is of note that if life emerged in an environment
containing both ribo- and 2′-deoxyribonucleotides, it is likely
that the first biopolymers were heterogeneous in composition,
that is, composed of a mixture of ribo- and deoxyribonucleo-
tides, and perhaps other compatible nucleotides. Recent work
has shown that functional biopolymers, in the form of aptamers
with highly specific molecular recognition properties, can be
derived by in vitro evolution from libraries of polynucleotides
composed of randomly interspersed ribo- and deoxyribo-
nucleotides. Thus, chimeric RNA/DNA polymers may have
been sufficient for the emergence of life. The primordial
biochemical exploitation of a mixed RNA and DNA genetic
system could eliminate the requirement for a genetic takeover
(of RNA by DNA), and would arguably result in a
simplification of the transition from chemistry to biology.
However, such mixed-composition polymers have neither the
advantageous stability of DNA, nor the optimal functional
characteristics of RNA. Thus, RNA and DNA may have
emerged as early and contemporaneous specializations of a
primitive mixed biopolymer.
SUMMARY
■
In summary, our results suggest that in a sulfur-rich prebiotic
environment, oxazole 1 and thiazole 7 could be formed
together under the same conditions, as long as both
glycolaldehyde 3 and its thiol analogue, β-mercapto-acetalde-
hyde 6, were present along with cyanamide 2. In such mixed
reactions, 2-aminothiazole 7 forms first, followed by 2-
aminooxazole 1. The absence of any interference between
oxazole and thiazole synthesis shows that these potential
precursors of deoxynucleotides and ribonucleotides could
indeed be formed at the same time in the same conditions.
We have also found that two abiotically plausible methods for
the purification of oxazole 1, namely, crystallization and
sublimation, also apply to thiazole 7. However, the chemical
reactivities of oxazole 1 and thiazole 7 are quite different,
suggesting that the subsequent pathways would diverge.
29
The most striking difference between the reactivity of oxazole
and thiazole 7 is the failure of the thiazole to react with
1
aldehydes to form the thiazoline precursors of the (thio)-
pyrimidine nucleotides. The most likely explanation for this
difference is the increased aromaticity of the thiazole relative to
the oxazole, and thus decreased nucleophilicity of C5. We are
currently investigating an alternative synthesis of 2′-deoxyribo-
pyrimidine nucleotides mediated by C2′-inversion of anhy-
EXPERIMENTAL SECTION
■
General Methods. Reagents and solvents were purchased from
Sigma-Aldrich, TCI America, Frontier Scientific, or Cambridge Isotope
Laboratories. Flash column chromatography was carried out on Merck
1a,11
dronucleotide 28 (Scheme 1).
However, during the course
9
385 silica gel 60 (230−400 mesh). NMR spectroscopy was carried
of efforts to observe two-component reactivity of thiazole 7
with aldehydes, we observed instead the crystallization of
glycolaldehyde 3 and homochiral (but not racemic) glycer-
out on a Varian NMR spectrometer (Oxford AS-400) or a Bruker
NMR spectrometer (AvanceIII 600) operating at 25 °C probe
temperature (unless otherwise specified). When possible, the chemical
shift of the corresponding solvent was used as a reference. Chemical
shift values are reported in parts per million (ppm) and J-couplings are
recorded in hertz (Hz). Electrospray mass spectrometry was recorded
on a Bruker Daltonics Esquire 6000 ESI-MS. High resolution mass
spectrometry was carried out on a Waters Q-ToF micro LC/MS/MS
system. Infrared spectra were recorded as manually pressed KBr discs
on a PerkinElmer spectrum 100 series FT-IR spectrometer. Single
crystal X-ray crystallography was carried out with a Bruker APEX II
CCD diffractometer (Mo Kα radiation, λ = 0.71073 Å), equipped with
an Oxford Cryosystems nitrogen flow apparatus, at 100 K. Data
1,3
aldehyde 22 (two known ribonucleotide precursors)
as
aminals directly from water. The facile reversibility of aminal
formation suggests that the chemical sequestration and
protection of C - and C -aldehyde nucleotide synthons by
2
3
another important class of nucleotide synthon (azoles) may
have provided a route to the concentration, purification, and
stabilization of the necessary aldehyde precursors of nucleo-
tides. Although these aldehydes can in principle be readily
2
6,27
synthesized by a variety of routes,
their high reactivity has
1
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Journal of the American Chemical Society
Article
integration down to 0.76 Å resolution was carried out using SAINT
V7.46 A (Bruker diffractometer, 2009) with reflection spot size
optimization. Absorption corrections were made with the program
SADABS (Bruker diffractometer, 2009). The structure was solved by
the direct methods procedure and refined by least-squares methods
NMR (101 MHz; DMSO): δ 166.6, 161.4, 141.6, 129.4, 111.4, 79.5,
C
+
48.0, 42.2. HRMS ESI [M + Na ] calc. for C H N OSNa 261.0535;
8
10
6
obs. 261.0526. Cambridge Crystallographic Data Centre deposition
number 864226.
18: Yield >85% d.r.= 2.5:1 erythro-18 (minor): R = 0.25 (CH Cl /
f
2
2
2
−1
against F by using SHELXS-97 and SHELXL-97 (Sheldrick, 2008).
Non-hydrogen atoms were refined anisotropically.
MeOH 7:3). IR (solid, cm ): 3437, 3387, 3292, 3123, 2975, 2856,
1
1638, 1613, 1571. H NMR (400 MHz; DMSO): δ 7.05 (s, 1H, Ar),
H
2
-Aminothiazole (7). Mercaptoacetaldehyde 6 and cyanamide 2
6.97 (s, 2H, NH ), 6.68 (br d, J = 19.9, 2H, NH ), 5.95 (d, J = 6.4, 1H,
2
2
(
0.5−1.5 equiv) were dissolved in H O, D O or phosphate buffer (1
H−(C1′)), 5.89 (s, 1H, HN), 4.54 (ddd, J = 6.4, 2.5, 1.7, 1H, H−
(C2′)), 3.78 (qd, J = 6.4, 2.6, 1H, H−(C3′)), 1.25 (d, J = 6.4, 3H, H−
2
2
1
mL) at pH/pD 7.0. Reaction progress was monitored by the H NMR
spectroscopy (see the Supporting Information, Figure S1). An initial
concentration of 100 mM led to direct crystallization of 7 (60%
isolated yield of crystals). Alternatively, upon complete conversion (by
NMR spectroscopy), the solution was concentrated and 7 recovered
by sublimation. Compound 7 (1−5 g) was placed in a covered beaker
or Erlenmeyer flask and warmed from beneath to 40 °C. Sublimate
was collected on flask/beaker sides and cover at ambient temperature
1
3
(C4′)). C NMR (101 MHz; DMSO): δ 166.7, 165.2, 141.4, 128.4,
c
+
111.6, 79.4, 58.6, 46.7, 22.7. HRMS ESI [M + Na ] calc. for
C H N OSNa 275.0691; obs. 275.0685. Cambridge Crystallographic
9
12
6
Data Centre deposition number 864227.
threo-18 (major): R = 0.25 (CH Cl /MeOH 7:3). IR (solid, cm ):
−
1
f
2
2
1
3391, 3302, 3103, 2972, 2865, 1651, 1621, 1582. H−NMR (400
MHz; DMSO): δ 7.33 (s, 1H), 6.96 (s, 1H), 6.76 (d, J = 27.2, 1H),
H
(
see Figure S4) or on a water-cooled coldfinger (5 °C). Crystalline 7
6
3
.20 (s, 1H), 6.00 (d, J = 6.1, 1H), 3.85 (dd, J = 9.5, 6.3, 1H), 3.24−
−1
.17 (m, 1H), 1.31 (d, J = 6.2, 1H). 13C NMR (101 MHz; DMSO): δ
was isolated. IR (solid, cm ): 3407, 3285, 3120, 3086, 1622, 1518,
c
1
1
488. H NMR (400 MHz; DMSO): δ 6.90 (d, J = 3.7, 1H, H−
H
166.7, 159.2, 141.3, 129.7, 111.3, 79.1, 54.6, 48.9, 20.4. HRMS ESI [M
13
(
(
(
C4)), 6.84 (s, 2H, NH ), 6.51 (d, J = 3.7, 1H, H−(C5)). C NMR
+
2
+ Na ] calc. for C H N OSNa 275.0691; obs. 275.0686. Cambridge
9
12
6
101 MHz; DMSO): δ 169.5, 139.3, 107.1. MS ESI (pos.) 101
C
Crystallographic Data Centre deposition number 864228.
9: Yield >85% d.r.= 3:1 R = 0.25 (CH Cl /MeOH 7:3). threo-19
+
100%, [M + H] ). Cambridge Crystallographic Data Centre
1
f
2
2
deposition number 864225.
−1
(
1
major): IR (solid, cm ): 3432, 3371, 3314, 3105, 2966, 2930, 1645,
604, 1575. H NMR (400 MHz; DMSO): δ 7.04 (s, 1H, Ar), 6.97
One-Pot Synthesis of 2-Aminothiazole (7) and 2-Amino-
oxazole (1). Glycolaldehyde 3 (1 equiv), mercaptoacetaldehyde 6 (1
equiv), and cyanamide 2 (0.5−5 equiv) were dissolved in H O/D O
1
H
(
5
s, 2H, NH ), 6.67 (br.s, 2H, NH ), 5.95 (d, J = 6.4, 1H, H−(C1′)),
2
2
2
2
.85 (s, 1H, NH), 4.60 (d, J = 6.3, 1H, H−(C2′)), 3.60 (t, J = 6.9, 1H,
(
1 mL) at pH/pD 7, 10 or 12, or 1 M phosphate buffer (1 mL) at pH/
H−(C3′)), 1.74−1.63 (m, 1H, H−(C4′H)), 1.46−1.35 (m, 1H, H−
1
pD 6.5−7.0. Reaction progress was monitored by the H NMR
spectroscopy (see the Supporting Information, Figure S8−10). An
initial concentration of 200 mM of 2, 3 and 6 led to direct
crystallization of 15 (5−10% isolated yield of crystals). Alternatively,
upon complete conversion (by NMR spectroscopy), the solution was
concentrated and 7 and 1 were recovered by sublimation. Compounds
13
(
1
C4′H)), 0.94 (t, J = 7.3, 3H, Me). C NMR (101 MHz; DMSO): δ
c
66.8, 165.3, 141.3, 128.3, 111.6, 79.6, 56.5, 52.5, 29.8, 10.6. HRMS
+
ESI [M + Na ] calc. for C H N OSNa 289.0848; obs. 289.0851.
10
14
6
Cambridge Crystallographic Data Centre deposition number 864229.
0: Yield >85% d.r.= 4:1 R = 0.2 (CH Cl /MeOH 7:3). threo-20
2
f
2
2
−1
(
1
major): IR (solid, cm ): 3404, 3109, 2923, 2525, 2293, 1605, 1581,
548, 1512. H NMR (400 MHz; DMSO): δ 6.70 (s, 1H, Ar), 5.58
7
and 1 (1:1; 100 mg) as fine powder or film (evaporated water) were
1
H
deposited in a covered glass tube (10 cm × 1 cm). A temperature
gradient (40, 50, or 60 °C to ambient temperature) was set up across
the tube and a mixture of crystalline thiazole 7 and oxazole 1 were
isolated at ambient temperature as a deposit upon the glass wall of the
tube.
(
=
d, J = 6.4, 1H, H−(C1′)), 4.28 (dd, J = 6.3, 2.1, 1H, NH)), 2.57 (dd, J
9.0, 2.2, 1H, H−(C2′)), 0.39 (qt, J = 8.1, 4.2, 1H, H−(C3′)), 0.18
quintet, J = 8.7, 2H, H −(C4′)), 0.00 (d, J = 3.9, 2H, H −(C4″)).
(
2
2
13
C NMR (101 MHz; DMSO): δ 164.4, 163.1, 138.8, 126.1, 109.2,
c
+
7
7.2, 54.9, 53.6, 15.1, 2.0, 0.0 HRMS ESI [M+Na ] calc. for
General Procedure for Isolation of Bis-thiazole Aminal
C H N OSNa 301.0846; obs. 301.0842. Cambridge Crystallographic
Crystals. Aldehyde (0.49−1.0 equiv) was added to a solution of 7
11 14
6
Data Centre deposition number 864230.
3: Yield >75% d.r.= 1:1. erythro-23 (major): R = 0.25 (CH Cl /
in H O at pH 7 ± 0.2. The solution was then incubated at room
2
2
temperature for 5−10 d; the resultant solid was then isolated by
filtration or trituration and washed twice with ethanol and once with
ice cold water. D-Glyceraldehyde 22/thiazole 7 crystals were then
f
2
2
−1
MeOH 7:3). IR (solid, cm ): 3363, 3213, 3192, 2920, 2844, 2375,
626, 1592, 1547, 1519. H NMR (400 MHz; D O): δ 7.23 (s, 1H,
1
1
2 H
Ar), 5.81 (d, J = 7.5, 1H, H−(C1′)), 4.99 (dd, J = 7.5, 2.5, 1H, H−
recrystallized from aqueous ethanol (10:1 H O/EtOH).
Glycolaldehyde Bis-thiazole Aminal 17. IR (solid, cm ): 3213
2
−1
(C2′)), 3.75 (ddd, J = 2.5, 5.9, 5.2 1H, H−(C3′)), 3.53 (ABX, J = 12.5,
1
5
(
.2, 1H, H−(C4′)), 3.49 (ABX, J = 12.1, 5.9, 1H, H−(C4′)). H NMR
(
br), 2966, 2934, 2869, 1589, 1539, 1497. Cambridge Crystallographic
400 MHz; DMSO-d ): δ 7.05 (s, 1H, Ar), 6.97 (s, 2H, CONH ),
Data Centre deposition number 864235.
Glyceraldehyde Bis-thiazole Aminal 19. IR (solid, cm ): 3326
6
H
2
−1
6.66 (bs, 2H, NH ), 6.05 (s, 1H, NH), 6.02 (d, J = 6.3, 1H, H−(C1′)),
2
5
1
.29 (t, J = 0.5, 1H, OH), 4.48 (d, J = 5.9, 1H, H−(C2′)), 3.78 (m,
H, H−(C3′)), 3.60 (m, 1H, H−(C4′)), 3.27 (m, 1H, H−(C4′)
(
br), 3244, 3113, 2948, 2933, 2901, 2882, 1536, 1489. Cambridge
Crystallographic Data Centre deposition number 864236.
1
3
partially obscured by DMSO residual solvent signal). C NMR (101
General Procedure for Multicomponent Reaction. Aldehyde
MHz; DMSO): δ 166.8, 165.9, 141.2, 128.3, 111.8, 80.2, 65.3, 53.6,
(
50−250 mM, 1 equiv) was added to a solution of 7 (1.1 equiv) and 5-
C
+
aminoimidazole-4-carboxamide 16 (1.5−3.0 equiv) in H O at pH 5.0
53.0. HRMS ESI [M + Na ] calc. for C
9
H
12
N
6
O
2
SNa 291.0640; obs.
2
±
0.2. The reaction was stirred under an argon atmosphere until no
291.0640. Cambridge Crystallographic Data Centre deposition
residual aldehyde was detectable following lyophilization and analysis
number 864231.
threo-23 (minor): R
1
−1
by H NMR. The reaction was lyophilized and purified by silica gel
f
= 0.20 (CH
2
Cl
2
/MeOH 7:3). IR (solid, cm ):
1
column chromatography (CH Cl /MeOH, 10:0 to 6:4). The major
3330, 3212, 3111, 2950, 2926, 2853, 2394, 1631, 1568. H NMR (400
MHz; D O): δ
2
2
product(s) were concentrated and recrystallized from ethanol/water or
2
H
7.35 (s, 1H, Ar), 5.98 (d, J = 5.9, 1H, H−(C1′)), 4.09
dichloromethane/methanol mixtures.
(dd, J = 7.9, 6.5, 1H, H−(C2′)), 3.80 (ABX, J = 12.0, 2.4, 1H, H−
−1
1
7: Yield 90% R = 0.25 (CH Cl /MeOH 7:3). IR (solid, cm ):
(C4′)), 3.62 (ABX, J = 11.9, 5.7, 1H, H−(C4′)), 3.45−3.41 (m, 1H,
f
2
2
1
1
3333, 3179, 2977, 2922, 2860, 1636, 1607, 1580, 1558. H NMR (400
H−(C3′)). H NMR (400 MHz; DMSO): δ
H
7.31 (s, 1H, Ar), 6.99
MHz; D O): δ 7.22 (s, 1H, Ar), 5.85 (d, J = 6.0, 1H, H−(C1′)), 4.18
(s, 2H, NH ), 6.76 (bd, J = 10.1, 2H, C(O)NH ), 6.35 (s, 1H, NH),
2
2
2
H
(
ddd, J = 7.8, 6.0, 4.1, 1H, H−(C2′)), 3.39 (ABX, J = 12.8, 4.1, 1H,
6.02 (d, J = 6.2, 1H, H−(C1′)), 5.21 (dd, J = 6.5, 4.6, 1H, HO), 3.95
(dd, J = 9.7, 6.2, 1H, H−(C2′)), 3.73 (dt, J = 11.1, 3.8, 1H, H−(C4′)),
1
H−(C3′)), 3.20 (ABX, J = 7.8, 1H, H−(C3′)). H NMR (400 MHz;
DMSO): δ 7.26 (d, J = 1.1, 1H, Ar), 5.91 (d, J = 6.1, 1H, H−(C1′)),
3.46−3.41 (m, inc. J = 11.1, 1H, H−(C4′) partially obscured by H O
H
2
13
4
3
.29−4.25 (ddd, J = 7.0, 6.1, 3.8, 1H, H−(C2′)), 3.41 (ABX, J = 13.0,
signal), 3.18 (m, inc. J = 7.2, 1H, H−(C3′)). C NMR (101 MHz;
DMSO): δ 166.5, 159.2, 140.9, 129.3, 111.0, 79.0, 63.2, 54.4, 49.0.
13
.8, 1H, H−(C3′)), 3.18 (ABX, J = 13.0, 7.0, 1H, H−(C3′)).
C
C
1
3893
dx.doi.org/10.1021/ja306176n | J. Am. Chem. Soc. 2012, 134, 13889−13895

Journal of the American Chemical Society
Article
+
HRMS ESI [M+H ] calc. for C H N O S 269.0821; obs. 269.0832.
(4) (a) Benner, S. A.; Ellington, A. D.; Tauer, A Proc. Nati. Acad. Sci.
U.S.A. 1989, 86, 7054. (b) Robertson, M. P.; Joyce, G. F. Cold Spring
Harb. Perspect. Biol. 2012, 4, a003608.
(5) (a) Reichard, P. Annu. Rev. Biochem. 1988, 57, 349. (b) Stubbe, J.
J. Biol. Chem. 1990, 256, 5329. (c) Stubbe, J. Curr. Opin. Struct. Biol.
6
13
6
2
Cambridge Crystallographic Data Centre deposition number 864232.
4: Yield >70% lyxo-24: R = 0.20 (CH Cl /MeOH 7:3). IR (solid,
cm ): 3344, 3106, 2917, 2833, 1627, 1553. H NMR (400 MHz;
2
f
2
2
−
1
1
DMSO): δ 7.14 (s, 1H, Ar), 6.90 (s, 2H, NH ), 6.59 (bs, 1H, NH ),
H
2
2
6
.35 (s, 1H, NH), 5.93 (d, J = 6.1, 1H, H−(C1′)), 4.98 (d, J = 6.1, 1H,
HO), 4.93 (t, J = 5.4, 1H, HO), 4.19 (dd, J = 7.6, 6.2, 1H, H−(C2′)),
.59−3.54 (m, 1H, H−(C4′)), 3.49 (ABX, J = 10.9, 5.0, 1H, H−
C5′)), 3.46−3.41 (m, inc. J = 10.9, 1H, H−(C5′)), 3.36−3.34 (m,
2
(
000, 10, 731.
6) Average bond enthalpies: C−S (255 kJ mol ) < C−O (355−
−
1
6a
−1
6b,c
3
3
1
3
80 kJ mol ).
̈
(a) Grelbig, T.; Potter, B.; Seppelt, K. Chem. Ber.
(
987, 120, 815. (b) Lovering, E. G.; Laider, K. J. Can. J. Chem. 1960,
8, 2367. (c) Levi, G. I.; Baladin, A. A. Bull. Acad. Sci. USSR, Div.
13
inc. J = 7.6 Hz, 1H, H−(C3′)). C NMR (100 MHz; DMSO): δC
1
5
3
66.6, 160.5, 148.7, 141.2, 128.6, 110.8, 78.38, 78.35, 71.2, 63.2, 53.3,
Chem. Sci. 1960, 149.
+
0.5. HRMS ESI [M + Na ] calc. for C H N O SNa 321.0746; obs.
21.0742. Cambridge Crystallographic Data Centre deposition
10
14
6
3
(
(
7) Schimpl, A.; Lemmon, R. M.; Calvin, M. Science 1965, 147, 149.
8) (a) Strecker, A. Liebigs Ann. Chem. 1850, 75, 27. (b) Miller, S. L.;
number 864233.
Urey, H. C. Science 1959, 130, 245.
9) (a) Sagan, C.; Khare, B. S. Science 1971, 173, 417. (b) Parker, E.
−
1
xylo-24: R = 0.20 (CH Cl /MeOH 7:3). IR (solid, cm ): 3309,
125, 2936, 2835, 1632, 1565. H NMR (400 MHz; D O): δ 7.26 (s,
f
2
2
(
1
3
2 H
T.; Cleaves, H. J.; Dworkin, J. P.; Glavin, D. P.; Callahan, M.; Aubrey,
A.; Lazcanoe, A.; Bada, J. L. Proc. Natl. Acad. Sci. U.S.A. 2011, 108,
1
H, Ar), 5.94 (d, J = 5.8, 1H, H−(C1′)), 4.21 (t, J = 6.7, 1H, H−
(
(
1
6
6
C2′)), 3.73 (m, 1H, H−(C4′)), 3.69 (ABX, J = 11.8, 4.5, 1H, H−
5
526.
C5′)), 3.64 (ABX, J = 11.8, 4.5, 1H, H−(C5′)), 3.52 (dd, J = 7.5, 1.9,
1
(10) Examples of desulfurization are legion in synthetic chemistry.
H, H−(C3′)). H NMR (400 MHz; DMSO): δ 7.05 (s, 1H, Ar),
H
(
a) Belen’kii, L. I. Chemistry of Organosulfur Compounds: Ellis
.99 (s, 1H, NH ), 6.67 (bs, 1H, NH ), 6.18 (s, 1H, NH), 6.01 (d, J =
2
2
Horwood, Chichester, 1990, p 193. (b) Larock, R. C. Comprehensive
Organic Transformations, 2nd ed.; Whiley-VCH: New York, 1999, pp
.3, 1H, H−(C1′)), 5.44 (d, J = 6.0, 1H, HO), 4.85 (t, J = 5.5, 1H,
HO), 4.55 (d, J = 6.1, 1H, H−(C2′)), 3.69 (d, J = 8.3, 1H, H−(C4′)),
5
3−60 and references therein. (c) Anderson, C. D.; Goodman, L.;
3
1
.54 (ABX(OH), J = 11.3, 5.5, 1H, H−(C5′)), 3.47 (ABX(OH), J =
13
Baker, B. R. J. Am. Chem. Soc. 1959, 81, 3967. (d) Uesugi, S.; Shida, T.;
Ikehara, M. Chem. Pharm. Bull. 1980, 28, 3621.
1.3, 5.5, 1H, H−(C5′)), 3.20 (m, 1H, H−(C3′)). C NMR (101
MHz; DMSO): δ 166.5, 165.9, 141.0, 127.9, 111.5, 100.0, 79.7, 75.1,
C
+
6
3.5, 53.5. HRMS ESI [M + Na ] calc. for C H N O SNa 321.0746;
obs. 321.0750. Cambridge Crystallographic Data Centre deposition
number 864234.
(11) C2′-inversion of ancitabine 28 (Scheme 1) by thiophosphate-
10
14
6
3
1
1a,b
11c
s
and the subsequent reduction of 2-thiocytidine have been
reported by Nagyvary and co-workers. However, Divakar et al.
questioned the validity of these results based upon the synthesis of
1
1d
ASSOCIATED CONTENT
Supporting Information
NMR and IR spectra, photographic images of sublimation
experiments, crystallographic information (CIFs) and a stereo-
chemical model for the addition of thiazole 7 to imine 21. This
material is available free of charge via the Internet at http://
pubs.acs.org.
similar compounds. (a) Bradbury, E.; Nagyvary, J. Nucleic Acids Res.
976, 3, 2437. (b) Patel, A. D.; Schrier, W. H.; Nagyvary, J. J. Org.
■
1
*
S
Chem. 1980, 45, 4830. (c) Patel, A. D.; Schrier, W. H.; Hrncir, M. A.;
Nagyvary, J. J. Origins Life 1981, 201. (d) Divakar, K. J.; Mottoh, A.;
Reese, C. B.; Sanghvi, Y. S. J. Chem. Soc., Perkin Trans. 1 1990, 969.
(
12) (a) Barone, R.; Chanon, M.; Gallo, R. Aminothiazoles and Their
Derivatives in The Chemistry of Heterocyclic Compounds, 2;. Vol.
4;Metzger, J. V., Ed.; Wiley: New York, 1979.
13) Currie, R. B.; Brutel, R.; Beutel, R. H. 1957, US PAT. 2,850,504.
3
(
AUTHOR INFORMATION
Corresponding Author
matthew.powner@ucl.ac.uk
(14) Significant H−(C4) exchange was observed in D O.
■
2
(
(
7
15) Caraoni, P. C.; Reboul, J. P. Acta Crystallogr. 1982, B38, 1255.
16) Cambridge Crystallographic Data Centre deposition number for
is 864225.
Notes
(
17) 16 is a product of HCN-oligomerization and known nucleobase
1
8
8
The authors declare no competing financial interest.
precursor. Notably, HCN also is required for Strecker synthesis.
18) (a) Oro, J. Biochem. Biophys. Res. Commun. 1960, 2, 407.
(b) Lowe, C. U.; Rees, M. W.; Markham, R. Nature 1963, 199, 219.
19) Cambridge Crystallographic Data Centre deposition numbers
for 17, erythro-18, threo-18, threo-19, and threo-20 are 864226, 864227,
64228, 864229, and 864230, respectively.
20) Proposed prebiotic nucleotide syntheses must be evaluated for
(
ACKNOWLEDGMENTS
■
(
M.W.P. was supported by a fellowship from the Harvard
Origins Initiative, the Howard Hughes Medical Institute and
University College London. This research was funded in part
by Grant CHE-0809413 from the NSF. J.W.S. is an Investigator
of the Howard Hughes Medical Institute. The authors thank
J.C. Blain, Dr. R.B. Moreno, Dr. A. Ricardo, and Prof. J.D.
Sutherland for helpful discussions and The Mass Spectrometry
Facility at the FAS Center for Systems Biology for mass
spectrometry support.
8
(
their potential to yield alternative structures capable of base-pairing.
Constitutionally, 3 (C ) and 22 (C ) can react with azoles (C sugar-
2
3
2
21
synthon) to furnish threose nucleic acid (TNA; C -sugar) and
DNA/RNA (C -sugar) precursors, respectively.
4
1
5
(
21) Scho
Eschenmoser, A. Science 2000, 290, 1347.
22) Cambridge Crystallographic Data Centre deposition numbers
for erythro-23, threo-23, lyxo-24, and xylo-24 are 864231, 864232,
64233, and 864234, respectively.
23) (a) Anastasi, C.; Crowe, M. A.; Powner, M. W.; Sutherland, J. D.
̈
ning, K.; Scholz, P.; Guntha, S.; Wu, X.; Krishnamurthy, R.;
(
REFERENCES
■
8
(
(
1) (a) Powner, M. W.; Gerland, B.; Sutherland, J. D. Nature 2009,
95, 239. (b) Powner, M. W.; Sutherland, J. D. Angew. Chem. 2010,
22, 4745; Angew. Chem., Int. Ed. 2010, 49, 4641. (c) Powner, M. W.;
Sutherland, J. D.; Szostak, J. W. Synlett 2011, 14, 1956. (d) Hein, J. E.;
Tse, E.; Blackmond, D. Nat. Chem. 2011, 3, 704.
4
1
Angew. Chem. 2006, 118, 6322; Angew. Chem., Int. Ed. 2006, 45, 6176.
(b) Sagi, V. S.; Karri, P.; Hu, F; Krishnamurthy, R. Angew. Chem. 2011,
123, 8277; Angew. Chem., Int. Ed. 2011, 50, 8127.
(
2) (a) Eschenmoser, A. Science 1999, 284, 2118. (b) The RNA
(24) The formation of aminal 25 was observed during the formation
of 2-aminothiazole 7 in presence of stoichiomtric glycolaldehyde 3 and
mercaptoacetaldehyde 6 and substiochiometric cyanamide 2.
(25) Cambridge Crystallographic Data Centre deposition numbers
for 25 and D-27 are 864235 and 864236, respectively.
World, 2nd ed.; Gesteland, R. F., Cech, T. R., Atkins, J. F., Eds.; Cold
Spring Harbor Laboratory Press: New York, 1999.
(
3) Powner, M. W.; Sutherland, J. D.; Szostak, J. W. J. Am. Chem. Soc.
2
010, 132, 16677. Erratum. J. Am. Chem. Soc. 2011, 133, 4149.
1
3894
dx.doi.org/10.1021/ja306176n | J. Am. Chem. Soc. 2012, 134, 13889−13895

Journal of the American Chemical Society
Article
(
26) (a) Butlerov, A. M. Acad. Sci. 1861, 53, 145. (b) Speck, J. C., Jr.
Adv. Carbohydr. Chem. Biochem. 1958, 13, 63. (c) Breslow, R.
Tetrahedron Lett. 1959, 21, 22. (d) Mizuno, T.; Weiss, A. H. Adv.
Carbohydr. Chem. Biochem. 1974, 29, 173. (e) Ahmed, S. N.; McKee,
M. L.; Shevlin, P. B. J. Am. Chem. Soc. 1985, 107, 1320. (f) Flanagan,
G.; Ahmed, S. N.; Shevlin, P. B. J. Am. Chem. Soc. 1992, 114, 3892.
(
g) Breslow, R.; Cheng, Z.-L. Proc. Natl. Acad. Sci. U.S.A. 2010, 107,
723. (h) Kim, H.-J.; Ricardo, A.; Illangkoon, H. I.; Kim, M. J.;
Carrigan, M. A.; Frye, F.; Benner, S. A. J. Am. Chem. Soc. 2011, 133,
457.
27) (a) Hollis, J. M.; Lovas, F. J.; Jewell, P. R. Astrophys. J. Lett.
000, 540, L107. (b) Halfen, D. T.; Apponi, A. J.; Woolf, N.; Polt, R.;
Ziurys, L. M. Astrophys. J. 2006, 639, 237.
28) N.B. C -thiazoline desulfurization would generate monovalent
5
9
(
2
(
4
nucleosides incapable of forming polymeric phosphodiesters.
29) Trevino, S. G.; Zhang, N.; Elenko, M. P.; Luptak, A.; Szostak, J.
W. Proc. Natl. Acad. Sci. U.S.A. 2011, 108, 13492.
(
1
3895
dx.doi.org/10.1021/ja306176n | J. Am. Chem. Soc. 2012, 134, 13889−13895