Tetrahedron 73 (2017) 4584e4590

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Tetrahedron
journal homepage: www.elsevier.com/locate/tet

Biocatalysed olefin reduction of 3-alkylidene oxindoles by baker's

yeast
Arianna Rossetti a, Alessandro Sacchetti a, b, *, Marta Bonfanti a, Gabriella Roda c,
Giulia Rainoldi d, Alessandra Silvani d
a
Dipartimento di Chimica, Materiali e Ingegneria Chimica “G. Natta”, Politecnico di Milano, P.zza Leonardo da Vinci 32, 20133 Milano, Italy
b
INSTM, National Consortium of Materials Science and Technology, Local Unit Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 Milan, Italy
c
Dipartimento di Scienze Farmaceutiche, Universita
degli Studi di Milano, Via Mangiagalli 25, 20133 Milano, Italy
d
degli Studi di Milano, Via C. Golgi 19, 20133 Milano, Italy
Dipartimento di Chimica, Universita

a r t i c l e i n f o a b s t r a c t

Article history: 3-Substituted oxindoles are very interesting molecules both for their potential biological activity and for
Received 15 March 2017 their role as starting materials toward more complex oxindole-based structures. These molecules can be
Received in revised form prepared by the reduction of a 3-ylidene oxindole precursor by classical metal-catalysed chemical re-
1 June 2017
ductions of the olefin. In this work we present a biocatalytic approach for the reduction of oxindole-
Accepted 13 June 2017
based olefins using baker's yeast. All the substrates were efficiently reduced in high yields. When an
Available online 16 June 2017
a,b-unsaturated ketone was used, the corresponding saturated alcohol was obtained in high yield and ee.
To further investigate the enzyme-substrate interactions a molecular docking study was also performed.
Keywords:
Biocatalysis
© 2017 Elsevier Ltd. All rights reserved.
Baker's yeast
Oxindole
Reduction
Molecular docking

1. Introduction interested in the metal-free reduction of 3-yliden-oxindole de-

rivatives by means of biocatalysis, in order to obtain 3-alkyl-oxin-
Oxindoles are present in many natural and synthetic products doles as possible substrates for further modifications. The use of
showing pharmaceutical and biological activities.1,2 Oxindole de- biocatalysis offers the advantage of mild reaction conditions with
rivatives have been employed as anti-cancer,3 antimicrobial,4,5 limited use of organic solvents. Conversions are usually very high
anti-HIV,6 and antimalarials.7,8 In the past we demonstrated the and when prochiral substrates are employed, chiral compounds can
utility of oxindoles as peptidomimetics capable of mimicking a b- be obtained in high enantiomeric excesses. Baker's yeast has been
turn structure.9,10 The most common derivatives are 3,3- demonstrated to be an efficient catalyst for the reduction of acti-
disubstituted oxindoles and 3-spirooxindoles containing a quater- vated olefins. This activity is due to the presence in this yeast of ene
nary carbon centre and diverse functional groups. Spirooxindoles reductase (ER) enzymes of the OYE family. The use of baker's yeast
are also related to a series of alkaloids that were first isolated from for chemical transformations is encouraged by the fact that re-
plants of the Apocynaceae11e13 and Rubiaceae14e17 families. actions are carried out in tap water, at room temperature and in the
The importance of isatin derivatives and their multiple appli- air in simple apparatus. The recovery of the products is easily
cations has stimulated the interest of the researchers. In the last realized by solvent extraction. An improvement of the recovery step
few years, lots of work has been devoted to finding an easy way to is possible when the substrates are supported onto insoluble
produce 3-substituted oxindoles, if possible with a controlled polystyrene resin, with the so-called Substrate Feeding Product
configuration at the stereocentre. Following our research activity in Removal (SFPR) technique. In accordance with our experience in
the preparation of oxindole-based compounds,18e21 we became the use of biocatalytic systems for the reduction of activated
olefins,22e26 we decided to investigate the baker's yeast mediated
reduction of olefins 1e13. Very recently, a similar approach using
* Corresponding author. Dipartimento di Chimica, Materiali e Ingegneria Chimica pseudomonas Monteilii was reported,27 thus confirming the urgent
“G. Natta”, Politecnico di Milano, P.zza Leonardo da Vinci 32, 20133 Milano, Italy. interest in this topic.28 Nevertheless, baker's yeast is more simple to
E-mail address: [email protected] (A. Sacchetti).

http://dx.doi.org/10.1016/j.tet.2017.06.022
0040-4020/© 2017 Elsevier Ltd. All rights reserved.
 A. Rossetti et al. / Tetrahedron 73 (2017) 4584e4590 4585

handle and scaling up to the preparation of grams of product is

easily achievable. We selected differently substituted olefins in
order to explore both the effect of different activating groups on the
reactivity and the role of steric hindrance.

2. Results and discussion

The substrates were synthesized according to Schemes 1 and 2.

Compounds 1e9 were prepared by standard Wittig olefination on
commercial isatins (see Scheme 3).
Compounds 10e12 were produced by the reaction of isatin with
the corresponding arylmethylketone with catalytic DEA (diethyl-
amine) followed by dehydration with thionyl chloride. The reaction
of isatin with ethylcyanoacetate and catalytic DBU (1,5-
diazabiciclo(5.4.0)undec-5-ene) afforded compound 13, while
grinding isatin with malononitrile and a substechiometric equiva-
lent of water afforded derivative 14.
The olefins were then submitted to reaction with dried yeast Scheme 2. Synthesis of compounds 10e14 by condensation reactions on isatin.
(Sigma-Aldrich) in tap water (150 g/L) in the presence of D-glucose
(15.0 g/L). Substrates were added at a concentration of 3 g/L. Re-
action were usually carried on the 100 mg scale of olefin. If
necessary, some drops of ethanol were added to facilitate the
dissolution of substrates. The mixtures were incubated at 35
C for
72 h in an orbital shaker. The same conditions were applied when
using the SFPR technique. In this case the resin (Amberlite XAD
1180-N) was loaded in a 1:20 substrate/resin ratio.
Results are reported in Table 1. Products 15e23 were isolated
from substrates 1e7, 13, 14 after chromatographic purification. As
expected when unsaturated ketones 8e12 were employed, the
corresponding saturated alcohols 24e28 were isolated as the major
product, due to the concomitant activity of the alcohol dehydro-
genase enzymes (ADH) which are present in the baker's yeast. The
saturated ketones 24a-28a were also obtained in a 2e6% yield,
according to 1H NMR analysis of the crude reaction mixture
(products were not isolated).
All the substrates could be efficiently transformed into products
Scheme 3. Reaction of olefins 1e14 with baker's yeast.
by baker's yeast. As expected, the use of the SFPR allowed isolation
of the final products in significantly higher yields. The lowest yields
were obtained with tetra-substituted olefins (21e23) probably due
as a result of their increased steric hindrance. Despite the use of of the C3 proton as a consequence of its exchange with deuterium
prochiral olefins, disappointingly no ee was detected after HPLC (see Fig. 1). These experiments proved the configurational lability of
analysis of the product 18. The preparation of enantioenriched 3- the C3 stereocenter due to the acidity of the hydrogen at C3.
alkyl 2-oxindoles has been previously reported in the literature Alcohols 25e28 were obtained as a 1:1 mixture of di-
by means of different approaches.29e33 In particular the reduction astereoisomers. In this case, as expected, the reduction of the
of 3-yliden-oxindoles catalysed by a chiral iridium complex was carbonyl group by ADH enzymes proved to be very enantiose-
reported to proceed with ee up to 93%.34 We reasoned that our lective, whereas the epimerisation at C3 led to no diastereoisomeric
results might be ascribable to an epimerisation of the newly formed excess. In fact, for representative compounds 24 and 25 an enan-
stereogenic centre.35 The chromatogram of the chiral HPLC sepa- tiomeric excess up to 98% could be measured by chiral HPLC. We did
ration of compound 18 is reported in Fig. 1. The presence of a not determine the absolute configuration at the alcohol stereo-
plateau between the peaks of the two enantiomers is characteristic center of the major enantiomer, however, according to the Prelog's
of a racemization process of the product during the column rule,37 we could suggest the stereochemical outcome proposed in
elution,36 due to epimerisation at the C3 position. To further Scheme 4. For compound 8, bearing the small methyl group, the S
investigate this aspect, the 1H NMR spectrum of 18 was first configuration can be safely assigned. Whereas in the presence of an
recorded in CDCl3. Upon addition of few drops of CD3OD, after aryl substituent, the opposite result should be expected.
24 h at room temperature we observed the complete disappearance According to the activation model for the reduction of activated
olefins by baker's yeast, as observed for substrates 6, 13 and 14, the
reaction suffers from the steric hindrance around the carbon-
carbon double bond. Despite the high steric demand introduced
by the oxindole moiety, for trisubstituted olefins the conversions
were good in all cases. Moreover, with the exception of compounds
1e3, two different activating EWGs are present on the double bond,
thus enabling different possible interaction modes with the ene-
reductase enzymes. To better understand these aspects of the
Scheme 1. Synthesis of 1e9 by Wittig olefination. substrate-enzyme interaction, we performed a study on the
4586 A. Rossetti et al. / Tetrahedron 73 (2017) 4584e4590

Table 1 model of the OYE 2 enzyme was created with the YASARA software
Reduction of substrates 1e14 by baker's yeast. and docking experiments were carried out with Autodock 4.2. Ac-
Substrate Product Yielda Yieldb drc cording to the mechanism, in our model the substrate is activated
1 15 71% 83% e
by interaction of the EWG of the olefin with His192 and Asp195
2 16 72% 77% e through hydrogen bonds. The olefin is then reduced by 1,4-addition
3 17 58% 79% e of hydride from FMNH, followed by protonation from Tyr197
4 18 68% 88% e (Fig. 2).
5 19 69% 78% e
We investigated the binding modes for representative com-
6 20 <5%c <5%c 60:40
7 21 67% 75% e pounds 1, 4, 8, and 14. The lowest energy poses for methylester 4
8 24 66% 82% 55:45 and olefin 1 are reported in Fig. 3. For 4 a normal mode of activation
9 25 62% 70% 50:50 is realized, with the coordination of the carboxymethyl group to the
10 26 59% 81% 52:48
active site. A further stabilizing hydrogen bond interaction between
11 27 44% 69% 57:43
12 28 51% 77% 51:49
the oxindole nitrogen and Tyr376 is detected. A similar situation is
13 22 e e e present for compound 8 (see SI). In compound 1 the activation is
14 23 14%c 13%c e still possible through the interaction with the oxindole carbonyl
a
Reaction performed without the use of resin. group, but with a different arrangement of the double bond, similar
b
With resin (SFPR). to the “flipped” mode previously reported with other substrates.
c
Calculated from 1H NMR and GC/MS analysis on the crude reaction mixture This result is noteworthy, since the oxindole ring forces an s-cis
(product not isolated).
conformation for the carbonyl-olefin system, a quite uncommon
geometry for the OYE reduction of olefins. On the contrary, the
docking pose obtained for 14 revealed a poor interaction of the
substrate. In fact, only one of the two cyano residues can form a
hydrogen bond and only with His192 (see SI). This result could be
an explanation of the poor reactivity of this substrate.

3. Conclusions

In this work we demonstrated the utility of baker's yeast for the

reduction of 3-ylidene oxindoles, aimed at the preparation of 3-
substituted oxindoles. Good results were obtained even with
highly hindered tri-substituted olefins bearing different functional
groups. No enantioselection was achieved, due to the easy race-
mization of the reduced product in the reaction medium. When a
keto group was present on the double bond, the diastereoisomeric
Fig. 1. Racemization process of compound 18. Chiral HPLC (left image) and 1H NMR saturated alcohols were obtained with very high enantiomeric
deuteration experiments are reported. excess.

Fig. 2. Activation mode for the OYE reduction of olefins.

Scheme 4. Possible stereochemical outcome of the ADH-mediated reduction, ac-

cording to the Prelog's rule.

docking modes of the oxindole-based olefins in an OYE 2 model

receptor. Though in baker's yeast different ene-reductase enzymes
are present, the activation of substrates and hydride transfer from
FMNH occur in similar way.38,39 Moreover, according to our expe- Fig. 3. Comparison of the binding modes of 4 (left) and 1 (right) in the active site of
rience, the stereochemical outcome of the reduction by baker yeast OYE2. Reducing cofactor FMNH is depicted in yellow.
or isolate enzymes is in most cases the same.24 The homology
 A. Rossetti et al. / Tetrahedron 73 (2017) 4584e4590 4587

4. Experimental section 4.5. Spectroscopic data for compounds 1e14

4.1. General 4.5.1. 3-Methyleneindolin-2-one1

1
H NMR (400 MHz, CDCl3) d 7.56 (s, 1H), 7.48 (d, J ¼ 7.3 Hz, 1H),
All solvents were distilled and dried over sodium or calcium 7.25 (d, J ¼ 7.7 Hz, 1H), 7.01 (d, J ¼ 7.7 Hz, 1H), 6.86 (d, J ¼ 7.6 Hz, 1H),
chloride, when necessary, prior to use. All chemicals were pur- 6.39 (s, 1H), 6.12 (s, 1H). 13C NMR (101 MHz, CDCl3) d 178.0, 142.3,
chased from commercial sources and used directly, unless indicated 134.9, 127.7, 125.4, 124.6, 122.4, 120.1, 110.3.
otherwise. All reactions were monitored by thin layer chromatog-
raphy (TLC) on precoated silica gel 60 F254; spots were visualized 4.5.2. 3-Ethylideneindolin-2-one2
with UV light (254 nm) or by treatment with 1% aqueous KMnO4 1
H NMR (400 MHz, DMSO-d6) d 10.38 (s, 1H), 7.48 (d, J ¼ 7.5 Hz,
solution. Products were purified by flash chromatography on silica 1H), 7.41 (t, J ¼ 7.5, 1H), 7.09 (q, J ¼ 6.4 Hz, 1H), 6.93 (t, J ¼ 7.4, 1H),
gel 60 (230e400 mesh). GCeMS analyses were performed by using 6.79 (d, J ¼ 7.5 Hz, 1H), 2.32 (d, J ¼ 6.4 Hz, 3H). 13C NMR (101 MHz,
an HP-5 MS column (30 m  0.25 mm  0.25 mm, Agilent). The DMSO-d6) d 168.3, 140.2, 137.0, 128.3, 127.7, 123.3, 120.8, 119.3,
following temperature program was employed: 60
C (1 min)/ 109.3, 13.7.
6
C min1/150
C (1 min)/12
C min1/280
C (5 min). The enan-
tiomeric excess was determined by HPLC on a Chiralpak AD column 4.5.3. 3-Benzylideneindolin-2-one3
(eluent 9:1, hexane: isopropanol, flow rate 0,75 mL/min). NMR 1
H NMR (400 MHz, CDCl3) d 8.29e8.25 (m, 2H), 7.78 (s, 1H),
spectra were recorded with a Bruker Avance 400 MHz spectrom- 7.57e7.51 (m, 2H), 7.48e7.42 (m, 3H), 7.22 (t, 1H, J ¼ 8.0 Hz), 7.04 (d,
eter. Chemical shifts (d) are expressed in ppm relative to TMS at 1H, J ¼ 7.6 Hz), 6.84 (t, 1H, J ¼ 8.0 Hz). 13C NMR (101 MHz, CDCl3)
d ¼ 0 ppm for 1H NMR and relative to CDCl3 at d ¼ 77.16 ppm for 13C d 167.5, 139.5, 137.6, 133.7, 132.0, 130.6, 128.9, 128.3, 126.1, 125.3,
NMR. Some 13C NMR spectra have been recorded using the APT 121.8, 119.3, 109.4.
pulse sequence; the signals of CH and CH3 are positive while CH2
and quaternary carbons are negative. FT-IR spectra were recorded 4.5.4. Methyl-2-(2-oxoindolin-3-ylidene)acetate4
in the ATR mode on an Agilent instrument. HR-EI mass spectra were 1
H NMR (400 MHz, CDCl3) d 8.57 (s, 1H), 8.53 (s, 1H), 7.28e7.36
measured on VG 70e70 EQeHF instrument equipped with its (m, 1H), 7.00e7.09 (m, 1H), 6.84e6.88 (m, 2H), 3.8 (s, 3H). 13C NMR
standard sources. (101 MHz, CDCl3) d 169.1, 165.9, 150.8, 132.6, 130.5, 129.2, 122.9,
122.0, 110.0, 52.1.
4.2. General procedure for the synthesis of compounds 1e3 by
wittig olefination
4.5.5. 2-(2-Oxoindolin-3-ylidene)acetonitrile5
1
H NMR (400 MHz, DMSO-d6) d 10.89 (s, 1H), 7.86 (d, J ¼ 7.6 Hz,
To a solution of the phosphonium salt (10.0 mmol) in THF
1H), 7.42 (dd, J ¼ 7.6, 7.6 Hz, 1H), 7.10 (dd, J ¼ 7.6, 7.6 Hz, 1H), 6.92 (d,
(70 mL) at 0
C, t-BuOK (1.26 g, 11.22 mmol) was added. The reac-
J ¼ 8.0 Hz, 1H), 6.53 (s, 1H). 13C NMR (101 MHz, DMSO-d6) d 166.9,
tion was stirred for 1 h to allow the formation of the ylide. A so-
145.6, 144.7, 134.8, 124.9, 123.3, 120.3, 117.8, 111.9, 98.6.
lution of isatin (10.0 mmol) in THF (70 mL) was added slowly to the
reaction mixture at 0
C. The resulting solution was stirred for
15 h at rt. The reaction was poured into saturated ammonium 4.5.6. Ethyl (Z)-2-(2-oxoindolin-3-ylidene)propanoate 6
chloride (70 mL) at 0
C. The compound was extracted with DCM Yellow solid. m.p. 180
C 1H NMR (400 MHz, CDCl3) d 8.74 (s, 1H),
(3  100 mL). The combined organic layers were dried (Na2SO4) and 7.58e7.44 (m, 1H), 7.24 (t, J ¼ 7.8 Hz, 1H), 7.02 (td, J ¼ 7.7, 1.1 Hz, 1H),
the solvent was removed under reduced pressure. The crude 6.85 (dt, J ¼ 7.7, 0.8 Hz, 1H), 4.40 (q, J ¼ 7.2 Hz, 2H), 2.43 (s, 3H), 1.39
product was purified by silica gel column chromatography eluting (t, J ¼ 7.1 Hz, 3H). 13C NMR (101 MHz, CDCl3) d 170.0, 169.3, 141.0,
with 2:8 EtOAc/hexane. 140.4, 129.8, 125.2, 122.9, 122.0, 121.1, 110.0, 61.8, 16.4, 14.1. nmax(-
ATR) 3079, 1726, 1703, 1613, 1467, 1113. HRMS (EI): calculated for
4.3. General procedure for the synthesis of compounds 4e9 by C13H13NO3: 231.0895; found 231.0894.
wittig olefination
4.5.7. Methyl (Z)-2-(5-methoxy-2-oxoindolin-3-ylidene)acetate 7
To a solution of isatin (0.04 mol) in DCM (150 mL), the corre- Brown solid. m.p. 184
C 1H NMR (400 MHz, CD3CN) d 8.37 (s,
sponding previously prepared stabilized ylide (0.04 mol) was 1H), 8.10 (d, J ¼ 2.7 Hz, 1H), 6.94 (dd, J ¼ 8.5, 2.7 Hz, 1H), 6.81 (d,
added. The mixture was stirred for 24 h at rt. The solution was J ¼ 8.5 Hz, 1H), 6.68 (s, 1H), 3.84 (s, 3H), 3.78 (s, 3H). 13C NMR
concentrated under reduced pressure and the residue purified by (101 MHz, CD3CN) d 178.4, 172.1, 156.1, 136.4, 131.1, 113.1, 111.7, 110.2,
silica gel column chromatography eluting with 2:8 EtOAc/hexane. 52.0, 42.9, 34.4, 14.2. nmax(ATR) 3175, 2922, 1702, 1437, 1207. HRMS
To provide the product as a solid. (EI): calculated for C12H11NO4: 233.0688; found 233.0692.

4.4. General procedure for the synthesis of compounds 10e12 4.5.8. (Z)-3-(2-oxopropylidene)indolin-2-one8
1
H NMR (400 MHz, CDCl3) d 8.50 (d, J ¼ 7.8 Hz, 1H), 7.84 (s, 1H),
Isatin (6 mmol) and the methylketone (6 mmol) were dissolved 7.32 (td, J ¼ 7.7, 1.3 Hz, 1H), 7.27e7.22 (m, 1H), 7.09e6.95 (m, 1H),
in 20 mL of ethanol. Diethylamine (124 mL, 1.2 mmol) was added 6.82 (d, J ¼ 7.8 Hz, 1H), 2.47 (d, J ¼ 1.1 Hz, 3H). 13C NMR (101 MHz,
and the reaction mixture was stirred at rt for 48 h. The solvent was CDCl3) d 198.55, 169.95, 155.76, 143.50, 135.63, 128.63, 127.87,
removed under reduced pressure and the residue was dissolved in 123.09, 120.75, 133.17, 32.35.
15 mL of DCM and SOCl2 (290 mL, 4 mmol) was added. After 2 h at rt
the solvent was removed under reduced pressure and the crude 4.5.9. (Z)-3-(2-oxo-2-phenylethylidene)indolin-2-one9
1
product was recrystallized from ethanol to afford the pure H NMR (400 MHz, CDCl3) d 8.34 (dt, J ¼ 7.8, 0.6 Hz, 1H),
compound. 8.18e8.05 (m, 2H), 7.89 (s, 1H), 7.71e7.63 (m, 1H), 7.59e7.50 (m,
Compounds 1340 and 1441 were prepared according to literature. 2H), 7.36e7.30 (m, 1H), 7.04 (td, J ¼ 7.7, 1.0 Hz, 1H), 6.87 (dd, J ¼ 7.8,
Spectroscopic data for compounds 1,42 2,43 3,44 4,45 5,46 8,47 9,48 1.1 Hz, 1H). 13C NMR (101 MHz, CDCl3) d 191.1, 169.4, 143.3, 137.6,
were in agreement with literature values. 136.7, 133.8, 132.7, 128.9, 128.8, 128.0, 126.5, 122.9, 120.7, 110.1.
4588 A. Rossetti et al. / Tetrahedron 73 (2017) 4584e4590

4.5.10. (Z)-3-(2-(4-chlorophenyl)-2-oxoethylidene)indolin-2-one 4.7.2. 3-Ethylindolin-2-one 16

1
10 H NMR (400 MHz, CDCl3) d 8.16 (br s, 1H), 7.23 (d, 1H,
Red solid. m.p. 225
C 1H NMR (400 MHz, DMSO-d6) d 10.79 (s, J ¼ 7.7 Hz), 7.22 (t, 1H, J ¼ 7.7 Hz), 7.04 (t, 1H, J ¼ 7.7 Hz), 6.88 (1H, d,
1H), 8.09 (d, J ¼ 8.6 Hz, 2H), 8.07e8.03 (m, 1H), 7.69 (s, 1H), 7.67 (d, J ¼ 7.7 Hz), 3.46 (1H, t, J ¼ 5.8 Hz), 2.04 (m, 2H), 0.93 (t, 3H,
J ¼ 8.6 Hz, 2H), 7.36 (td, J ¼ 7.7, 1.3 Hz, 1H), 6.97 (td, J ¼ 7.7, 1.1 Hz, J ¼ 7.5 Hz). 13C NMR (101 MHz, CDCl3) d 180.1, 141.6, 129.5, 127.8,
1H), 6.89 (dt, J ¼ 7.8, 0.8 Hz, 1H). 13C NMR (101 MHz, DMSO-d6) 124.1, 122.2, 109.5, 47.0, 23.5, 9.9.
d 190.6, 168.6, 145.5, 139.4, 137.3, 136.2, 133.6, 131.0, 129.7, 127.3,
125.8, 122.2, 120.4, 110.9. nmax(ATR) 3146, 3085, 1715, 1495, 1327. 4.7.3. 3-Benzylindolin-2-one 17
1
HRMS (EI): calculated for C16H10ClNO2: 283.0400; found 283.0404. H NMR (400 MHz, CDCl3) d 9.32 (1H, br s), 7.29e7.15 (6H, m),
7.03 (1H, td, J ¼ 7.5, 1.0 Hz), 6.92e6.86 (2H, m); 6.75 (1H, d,
4.5.11. (Z)-3-(2-(4-methoxyphenyl)-2-oxoethylidene)indolin-2-one J ¼ 7.4 Hz), 3.77 (1H, dd, J ¼ 9.3, 4.5 Hz), 3.52 (1H, dd, J ¼ 13.7,
11 4.6 Hz), 2.95 (1H, dd, J ¼ 13.7, 9.3 Hz); 13C NMR (101 MHz, CDCl3)
Brown solid. m.p. 197
C 1H NMR (400 MHz, DMSO-d6) d 10.76 (s, d 180.1, 141.5, 137.8, 129.4, 129.0, 128.3, 127.9, 126.6, 124.7, 121.9,
1H), 8.06 (d, J ¼ 8.9 Hz, 2H), 7.94 (d, J ¼ 7.7 Hz, 1H), 7.69 (s, 1H), 7.33 109.8, 47.6, 36.6.
(td, J ¼ 7.7, 1.3 Hz, 1H), 7.13 (d, J ¼ 8.9 Hz, 2H), 6.94 (td, J ¼ 7.7, 1.3 Hz,
1H), 6.88 (dt, J ¼ 7.8, 0.8 Hz, 1H), 3.88 (s, 3H). 13C NMR (101 MHz, 4.7.4. Methyl 2-(2-oxoindolin-3-yl)acetate 18
1
DMSO-d6) d 190.2, 168.7, 164.4, 145.1, 136.0, 133.0, 131.6, 130.4, 127.1, H NMR (400 MHz, CDCl3) d 8.88 (br s, 1H), 7.24e7.20 (m, 2H),
127.0, 122.1, 120.5, 114.9, 110.8, 56.2. nmax(ATR) 3147, 1706, 1603, 7.01 (d, J ¼ 7.6 Hz, 1H), 6.91 (d, J ¼ 8.0 Hz, 1H), 3.82 (dd, J ¼ 4.8,
1457, 1010. HRMS (EI): calculated for C17H13NO3: 279.0895; found 8.0 Hz, 1H), 3.70 (s, 3H), 3.09 (dd, J ¼ 4.4, 16.8 Hz, 1H), 2.84 (dd,
279.0893. J ¼ 8.0, 16.8 Hz, 1H). 13C NMR (101 MHz, CDCl3) d 179.2, 171.6, 141.5,
128.6, 128.3, 124.1, 122.5, 109.9, 52.1, 42.3, 34.5.
4.5.12. (Z)-3-(2-(furan-2-yl)-2-oxoethylidene)indolin-2-one 12
Red solid. m.p. 188
C 1H NMR (400 MHz, DMSO-d6) d 10.80 (s, 4.7.5. 2-(2-Oxoindolin-3-yl)acetonitrile 19
1
1H), 8.48 (d, J ¼ 7.8 Hz, 1H), 8.14 (d, J ¼ 1.0 Hz, 1H), 7.71 (dd, J ¼ 3.7, H NMR (400 MHz, DMSO-d6) d 10.62 (1H, s), 7.39 (1H, d,
1.0 Hz, 1H), 7.59 (s, 1H), 7.39 (td, J ¼ 7.7, 1.0 Hz, 1H), 7.03 (dd, J ¼ 7.6, J ¼ 7.4 Hz), 7.24 (1H, t, J ¼ 7.7 Hz), 7.01 (1H, t, J ¼ 7.5 Hz), 6.88 (1H, d,
1.1 Hz, 1H), 6.89 (d, J ¼ 8.1 Hz, 1H), 6.83 (dd, J ¼ 3.7, 1.7 Hz, 1H). 13C J ¼ 7.7 Hz), 3.82 (1H, t, J ¼ 5.9 Hz), 3.20 (1H, dd, J ¼ 17.2, 5.8 Hz), 3.06
NMR (101 MHz, DMSO-d6) d 177.2, 168.7, 153.8, 149.6, 145.8, 133.2, (1H, dd, J ¼ 17.2, 5.9 Hz). 13C NMR (101 MHz, DMSO-d6) d 176.4,
128.6, 128.0, 126.0, 124.2, 122.3, 120.6, 113.9, 110.8. nmax(ATR) 3186, 142.8, 128.6, 127.1, 124.2, 121.6, 118.2, 109.6, 41.3, 17.6.
2921, 1706, 1458, 1328. HRMS (EI): calculated for C14H9NO3:
239.0582; found 239.0581. 4.7.6. Methyl 2-(5-methoxy-2-oxoindolin-3-yl)acetate 21
Orange solid. m.p. decompose. 1H NMR (400 MHz, CD3CN) d 8.31
4.5.13. Ethyl-2-cyano-2-(2-oxoindolin-3-ylidene)acetate 13 (s, 1H), 6.90e6.70 (m, 3H), 3.76 (s, 3H), 3.70e3.65 (m, 1H), 3.64 (s,
1
H NMR (400 MHz, CDCl3) d 7.94 (d, J ¼ 8.0 Hz, 1H), 7.56 (s, 1H), 3H), 3.00 (dd, J ¼ 16.8, 5.3 Hz, 1H), 2.86 (dd, J ¼ 17.0, 6.9 Hz, 1H). 13C
7.13 (t, J ¼ 7.7 Hz, 1H), 6.71 (t, J ¼ 7.8 Hz, 1H), 6.61 (d, J ¼ 8.3 Hz, 1H), NMR (101 MHz, CD3CN) d 171.5, 166.6, 155.9, 139.4, 138.9, 123.1,
4.21 (q, J ¼ 7.2 Hz, 2H), 1.19 (t, J ¼ 7.1 Hz, 3H). 13C NMR (101 MHz, 121.7, 119.2, 114.6, 111.3, 56.0, 52.5. nmax(ATR) 3263, 2924, 1711, 1487,
CDCl3) d 165.2, 164.5, 163.8, 135.9, 127.9, 126.8, 126.4, 124.0, 120.3, 1203. HRMS (EI): calculated for C12H13NO4: 235.0845; found
117.2, 106.3, 59.1, 13.7. 235.0841.

4.5.14. 2-(2-Oxoindolin-3-ylidene)malononitrile14 4.7.7. 3-(2-hydroxypropyl)indolin-2-one 24. Mixture of two

1
H NMR (400 MHz, DMSO-d6) d 11.21 (s, 1H), 7.89 (d, J ¼ 8.0 Hz, diastereoisomers
1H), 7.57 (dd, J ¼ 8.0, 7.6 Hz, 1H), 7.14 (dd, J ¼ 8.0, 7.6 Hz, 1H), 6.93 (d, Yellow solid. m.p. 84
C (decompose) 1H NMR (400 MHz, CDCl3)
J ¼ 8.0 Hz, 1H). 13C NMR (101 MHz, DMSO-d6) d 164.7, 151.5, 147.4, d 9.01 (s, 0.5H, diast A), 8.80 (s, 0.5H, diast B), 7.25e7.13 (m, 2H),
138.7, 126.7, 123.8, 119.6, 114.0, 112.5, 112.4, 81.5. 7.09e6.98 (m, 1H), 6.93e6.84 (m, 1H), 4.24 (td, J ¼ 6.4, 3.0 Hz, 0.5H,
diast A), 4.17e4.08 (m, 0.5H, diast B), 3.73e3.60 (m, 1H), 2.89 (d,
4.6. General procedure for the baker's yeast reduction of olefins J ¼ 6.3 Hz, 1H), 2.16 (ddd, J ¼ 14.0, 9.1, 4.7 Hz, 0.5H, diast A),
2.04e1.82 (m, 1.5H), 1.30 (d, J ¼ 6.2 Hz, 1.5H, diast A), 1.27 (d,
To a mixture of commercial fresh or freeze-dried baker's yeast J ¼ 6.3 Hz, 1.5H, diast B). 13C NMR (101 MHz, Chloroform-d) d 181.7
(Sigma) (7.5 g) in tap water (50 mL) at 40
C, D-glucose (0.75 g, and 181.6, 141.3 and 141.1, 129.4 and 129.3, 127.1 and 126.8, 123.9
4.17 mmol) was added. The substrate (100 mg) was added and the and 123.8, 122.7 and 122.5, 110.1 and 109.9, 64.4 and 64.2, 44.7 and
flask was placed for 2 d in a termo-shaker. The mixture was then 44.2, 41.1 and 40.0, 23.6 and 23.3. nmax(ATR) 3437, 2925, 2987, 1470,
filtered on a celite pad and washed twice with water (50 mL) and 1137. HRMS (EI): calculated for C11H13NO2: 191.0946; found
once with EtOAc (50 mL). The aqueous was extracted with EtOAc 191.0944.
(3  50 mL). The combined organic layers were dried (Na2SO4) and
the solvent was removed under reduced pressure. The crude res- 4.7.8. 3-(2-Hydroxy-2-phenylethyl)indolin-2-one 25
idue was purified by silica gel column chromatography eluting with Yellow solid. m.p. 104
C mixture of two diastereoisomers. 1H
4:6 EtOAc/hexane to give the product. NMR (400 MHz, CDCl3) d 9.03 (s, 0.5H, diast A), 8.90 (s, 0.5H, diast B),
7.38e6.98 (m, 8H), 6.91e6.80 (m, 1H), 5.12e5.03 (m, 1H), 4.74 (s,
4.7. Spectroscopic data for compounds 15e17,48 and 18, 1949 were 0.5H, diast A), 4.17 (s, 0.5H, diast B), 3.69 (t, J ¼ 7.0 Hz, 0.5H, diast A),
in agreement with literature values 3.59 (dd, J ¼ 9.3, 3.9 Hz, 0.5H, diast B), 2.40 (ddd, J ¼ 14.6, 8.1, 4.0 Hz,
0.5H, diast A), 2.27e2.10 (m, 1.5H). 13C NMR (101 MHz, CDCl3)
4.7.1. 3-Methylindolin-2-one 15 d 181.7 and 181.6, 142.8 and 142.6, 141.3 and 141.1, 133.2 and 133.0,
1
H NMR (400 MHz, CDCl3) d 9.23 (1H, br s), 7.21 (2H, td, J ¼ 7.4, 129.4 and 129.3, 128.8e128.6 (2C), 128.3 and 128.1, 127.3 and 127.0
0.8 Hz), 7.03 (1H, td, J ¼ 7.5, 1.0 Hz), 6.93 (1H, d, J ¼ 7.5 Hz), 3.47 (1H, (2C), 124.0 and 123.8, 122.9 and 122.7, 110.2 and 110.1, 72.8 and 70.5,
q, J ¼ 7.7 Hz), 1.51 (3H, d, J ¼ 7.7 Hz). 13C NMR (101 MHz, CDCl3) 45.3 and 43.0, 40.3 and 39.0. nmax(ATR) 3250, 1698, 1620, 1470.
d 181.4, 141.2, 131.2, 127.9, 123.8, 122.3, 109.7, 41.0, 15.2. HRMS (EI): calculated for C16H15NO2: 253.1103; found 253.1106.
 A. Rossetti et al. / Tetrahedron 73 (2017) 4584e4590 4589

4.7.9. 3-(2-(4-chlorophenyl)-2-hydroxyethyl)indolin-2-one 26 negative energies mean no binding). After clustering the 50 runs,
Yellow solid. m.p. 125
C mixture of two diastereoisomers. 1H the resulting complex conformations were originated (they all
NMR (400 MHz, CDCl3) d 9.03 (s, 0.5H, diast A), 8.90 (s, 0.5H, diast B), differ by at least 5.0 A heavy atom RMSD). Binding energy are re-
7.36e7.25 (m, 4H, diast Aþ diast B), 7.22e7.10 (m, 2H, diast Aþ diast ported in kcal/mol and predicted dissociation constant in pM units.
B), 7.06e6.96 (m, 1H, diast Aþ diast B), 6.90e6.83 (m, 1H, diast Aþ Contacting receptor residues are also listed. The poses were then
diast B), 5.15e5.01 (m, J ¼ 7.8, 5.4 Hz, 1H, diast Aþ diast B), 4.74 (s, viewed using PyMOL.52
0.5H, diast A), 4.17 (s, 0.5H, diast B), 3.69 (t, J ¼ 6.9 Hz, 0.5H, diast A),
3.59 (dd, J ¼ 9.3, 3.9 Hz, 0.5H, diast B), 2.40 (ddd, J ¼ 14.6, 8.1, 4.0 Hz, Appendix A. Supplementary data
0.5H, diast A), 2.26e2.10 (m, 1.5H, diast Aþ diast B). 13C NMR
(101 MHz, CDCl3) d 181.7 and 181.6, 142.8 and 142.6, 141.3 and 141.1, Supplementary data related to this article can be found at http://
133.2 and 133.0, 129.4 and 129.3, 128.6 and 128.6 (2C), 128.3 and dx.doi.org/10.1016/j.tet.2017.06.022.
128.1, 127.3 and 127.0 (2C), 124.0 and 123.8, 122.9 and 122.7, 110.2
and 110.1, 72.8 and 70.5, 45.3 and 43.0, 40.3 and 39.0. nmax(ATR)
3195, 1687, 1613, 1510, 1469, 1247. HRMS (EI): calculated for References
C16H14ClNO2: 287.0713; found 287.0715.
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Yellow solid. m.p. 115
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Yellow solid. m.p. 101
C (decompose) mixture of two diaster- 16. Sakakibara I, Takahashi H, Terabayashi S, et al. Phytomedicine. 1998;5(2):
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4537e4541.
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