Synthesis of 4-substituted 3-[3-(dialkylaminomethyl)indol-1-yl]maleimides and study of their ability to inhibit protein kinase C-α, prevent development of multiple drug resistance of tumor cells and cytotoxicity

Article abstract of DOI:10.1007/s11172-008-0271-9

A series of 3-[3-(dialkylaminomethyl)indol-1-yl]maleimides containing the indole, dihydroindole, mercaptophenol, or tetrahydroquinoline residues at position 4 of the maleimide ring, as well as 3-(dialkylaminomethyl)indole derivatives have been synthesized. Their ability to inhibit in vitro protein kinase C-α (PKC-α) has been studied. Cytotoxicity of new compounds and their ability to constrain activation of multiple drug resistance (MDR) have been studied in the human tumor cell line. Both the toxic and the low-toxic PKC-α inhibitors prevent the activation of MDR in the tumor cells. Among compounds under study, a number of substances have been found that prevent the activation of MDR but do not inhibit PKC-α.

Full text of DOI:10.1007/s11172-008-0271-9

Russian Chemical Bulletin, International Edition, Vol. 57, No. 9, pp. 2011—2020, September, 2008
2011
Synthesis of 4ꢀsubstituted 3ꢀ[3ꢀ(dialkylaminomethyl)indolꢀ
1ꢀyl]maleimides and study of their ability to inhibit
protein kinase Cꢀα, prevent development of multiple
drug resistance of tumor cells and cytotoxicity
A. Yu. Simonov,a S. A. Lakatosh,a Yu. N. Luzikov,a M. I. Reznikova,a O. Yu. Susova,b A. A. Shtil',b
S. M. Elizarov,c V. N. Danilenko,d and M. N. Preobrazhenskayaa
aG. F. Gause Research Institute of New Antibiotics, Russian Academy of Medical Sciences,
11 ul. B. Pirogovskaya, 119021 Moscow, Russian Federation.
Fax: +7 (499) 245 0295. Eꢀmail: mnp@space.ru
bN. N. Blokhin Russian Cancer Scientific Center, Russian Academy of Medical Sciences,
24 Kashirskoe sh., 115478 Moscow, Russian Federation.
Fax: +7 (495) 324 1114
cA. N. Bakh Institute of Biochemistry, Russian Academy of Sciences,
33, str. 2 Leninsky prosp., 119071 Moscow, Russian Federation.
Fax: +7 (495) 954 2732
dN. I. Vavilov Institute of General Genetics, Russian Academy of Sciences,
3 ul. Gubkina, 119991 Moscow, Russian Federation.
Fax: +7 (495) 132 8962
A series of 3ꢀ[3ꢀ(dialkylaminomethyl)indolꢀ1ꢀyl]maleimides containing the indole, dihyꢀ
droindole, mercaptophenol, or tetrahydroquinoline residues at position 4 of the maleimide ring,
as well as 3ꢀ(dialkylaminomethyl)indole derivatives have been synthesized. Their ability to inhibit
in vitro protein kinase Cꢀα (PKCꢀα) has been studied. Cytotoxicity of new compounds and their
ability to constrain activation of multiple drug resistance (MDR) have been studied in the
human tumor cell line. Both the toxic and the lowꢀtoxic PKCꢀα inhibitors prevent the activaꢀ
tion of MDR in the tumor cells. Among compounds under study, a number of substances have
been found that prevent the activation of MDR but do not inhibit PKCꢀα.
Key words: bisindolylmaleimides, protein kinase C, targetꢀspecific therapy, antitumor medꢀ
icines, transformed cells, cytotoxicity, multiple drug resistance.
Currently, the search for inhibitors of protein kinases
became one of important directions in the quest for new
medicinal agents.1 Protein kinases C (PKC) is a family of
serineꢀthreonine protein kinases playing a key role in the
regulation of cellular processes in eukaryotes. The involveꢀ
ment of some PKC isoforms, in particular, the αꢀisoform
(PKCꢀα) in the survival of tumor cells is especially imporꢀ
tant.1,2 It was found that it mediates the intracellular sigꢀ
nals regulating expression of the gene MDR1 (Multidrug
Resistance 1). The product of this gene, the transmemꢀ
brane protein Рꢀglycoprotein, participating in the active
outbreak of chemotherapeutic drugs from the cells into the
intercellular medium, is the factor inducing multiple drug
resistance (MDR) in tumor cells.3,4 The gene MDR1 can
be activated by the action of antitumor drugs. The acute
(during the first hours of action) activation of MDR1 leads
to the accumulation of Рꢀglycoprotein and development
of MDR.5 Since the activation of MDR1 and MDR itself
are mediated by PKCꢀα, the inhibition of this isoform
prevents the emergence of MDR. In fact, bis(indolyl)ꢀ
maleimide I, which is an inhibitor of PKCꢀα, prevents the
activation of MDR1 by antitumor drugs.6
Bis(indolꢀ3ꢀyl)maleimides, which nowadays are under
various stages of clinical trials, belong to inhibitors of proꢀ
tein kinases.1,7 Among bis(indolꢀ3ꢀyl)maleimides and reꢀ
lated fused compounds, e.g., derivatives of indolocarbꢀ
azole such as Rebeccamicin and Staurosporin, the highly
active inhibitors of serineꢀthreonine protein kinases and
topoisomerase I, triggering tumor cell apoptosis, have been
found.1,8 Compounds BisꢀI and BisꢀIII are wellꢀknown
PKC inhibitors, they derive from unsubstituted bis(indolꢀ
3ꢀyl)maleimide (BisꢀIV), which is the inhibitor of PKC,
too. The active derivatives contain the maleimide core unꢀ
substituted at the nitrogen atom and aminoalkyl substituꢀ
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 9, pp. 1977—1985, September, 2008.
1066ꢀ5285/08/5709ꢀ2011 © 2008 Springer Science+Business Media, Inc.

2012
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
Simonov et al.
ent at position 1 of one of the indole rings in which the
substituted or unsubstituted amino group is separated from
the indole core by three carbon atoms.8
fore, it can be assumed that new efficient inhibitors of PKC
can be designed on their basis.
Results and Discussion
In the present work, a series of heterocycles 1—7 were
converted into derivatives containing 3ꢀdialkylaminomeꢀ
thyl group in the indole ring (Scheme 1), and their properꢀ
ties were assessed aimed at the evaluation of their prosꢀ
pects for the use in oncology.
The Mannich reaction with 3,4ꢀbis(indolꢀ1ꢀyl)ꢀ1ꢀmeꢀ
thylmaleimide (5) gave rise to monoꢀ and bisaminoꢀ
methylation products, which were separated by chromaꢀ
tography.
In contrast to the known PKC inhibitors, the indole
fragment is bound in the compounds under study to
the maleimide ring through the nitrogen atom, whereas
the dialkylamino group is separated from the indole ring by
one carbon atom. However, the distances between the niꢀ
trogen atoms of the side chain and the indole ring in comꢀ
pound BisꢀI and in the aminomethyl derivatives obtained
are close and are equal to 4.64 and 4.57 Å, respectively.11
For the new compounds, their ability to inhibit PKCꢀα
a cellꢀfree test system, as well as cytotoxicity in the human
leucosis cell line, and the ability to prevent activation of
gene MDR1 under the action of antitumor drug Cytosar
(cytosine βꢀDꢀarabinofuranoside) have been investigated.
A relationship between the amount of radioactive phosꢀ
phate transferred by the kinase from [γꢀ32P]ATP to its
protein substrate and the concentration of the inhibitor
has been studied. The cytotoxicity is expressed as InC50
(inhibitory concentration that causes death of 50% of cells
R = Me (BisꢀI), H (BisꢀIII)
Earlier,9,10 methods for the preparation of 3,4ꢀbisꢀ
(indolꢀ1ꢀyl)maleimides and 3ꢀ(indolꢀ1ꢀyl)maleimide deꢀ
rivatives have been developed. These compounds are the
structural analogs of 3,4ꢀbis(indolꢀ3ꢀyl)maleimides, thereꢀ
1: R = H (a), Me (b), Et (c), (CH2)3NMe2 (d)
6: Y = H, X = Cl
7: Y = COOMe, X = H

3ꢀ(Dialkylaminomethylindolꢀ1ꢀyl)maleimides
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
2013
Scheme 1
8: Nu = indolinꢀ1ꢀyl, R = H, R´ = Et (a); R = H, NR´2 = pyrrolidino (b); R = R´ = Me (c); R = Me, NR´2 = pyrrolidino (d);
R = Me, NR´2 = morpholino (e); R = Me, NR´ = 4ꢀmethylpiperazino (f); R = R´ = Et (g); R = —(CH2)3NMe2, R´ = Et (h)
9: Nu = 4ꢀmethoxyphenylthio, R = Me; NR´2 = NMe2 (a), NEt2 (b), pyrrolidino (c), morpholino (d), 4ꢀmethylpiperazino (e)
10: Nu = Nꢀethylanilino, R = Me; NR´2 = NMe2 (a), NEt2 (b), pyrrolidino (c), morpholino (d), 4ꢀmethylpiperazino (e)
11: Nu = 6ꢀmethylꢀ1,2,3,4ꢀtetrahydroquinolino, R = Me, R´ = Et
12: Nu = 5ꢀchloroindolino, R = Me, R´ = Et
13: Nu =
, R = Me, R´ = Et
14: NR2 = NMe2 (a), NEt2 (b), pyrrolidino (c), morpholino (d), 4ꢀmethylpiperazino (e)
15: NR2 = NMe2 (a), pyrrolidino (b), morpholino (c), 4ꢀmethylpiperazino (d)
after 72 h of incubation) for the К562 leucosis cells line.
Compounds with InC50 < 10 μmol L–1 were considered as
cytotoxic.
Prevention of MDR was determined from the decrease
in activation of the gene MDR1 in the К562 cells treated
with Cytosar (10 μmol L–1, 16 h) in combination with
a compound under study (PC50).5,6
The main part of compounds synthesized belongs to this
group. Lowꢀtoxic compounds (**) are less numerous and
are mainly represented by 3ꢀ(2,3ꢀdihydroindolꢀ1ꢀyl)ꢀ4ꢀ
(indolꢀ1ꢀyl)ꢀ1ꢀmethylmaleimide derivatives (compounds
8c—f). It is essential that all of them suppress induction of
MDR and are potent PKCꢀα inhibitors. Some of them
exhibit activity comparable with, or exceeding, the activity
of the standard inhibitor BisꢀI. Monoꢀ and bis(dialꢀ
kylaminomethyl) derivatives of 3,4ꢀbis(indolꢀ1ꢀyl)ꢀ1ꢀmeꢀ
thylmaleimide possessed high cytotoxicity (***).
A considerable difference of PKCꢀα inhibitors obtained
from the known bis(indolꢀ3ꢀyl)maleimide derivatives
should be emphasized: the introduction of alkyl (amiꢀ
noalkyl) substituents at the maleimide nitrogen atom has
no noticeable effect on the activity, which follows, for
example, from the comparison of compounds 8a and 8g.
In the case of bis(indolꢀ3ꢀyl)maleimide derivatives, subꢀ
The data obtained are summarized in Table 1.
From the data given in Table 1, it follows that the
overwhelming part of compounds synthesized are capable
of suppressing the MDR induction at concentrations comꢀ
parable or less than the concentration of BisꢀI, the stanꢀ
dard PKCꢀα inhibitor. Compounds 8b, 9b, and 9c are the
exceptions, they are efficient at higher concentrations.
Compounds under study can be divided into three
groups with regard to their cytotoxicity. Moderately cytoꢀ
toxic compounds are marked with an asterisk (*) in Table 1.

2014
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
Simonov et al.
Table 1. Biological properties of maleimide deꢀ
rivatives
In addition, compounds 10c, 10e, and 14a are not PKCꢀα
inhibitors being at the same time efficient suppressors of
the MDR induction. This allows one to suppose that not
only PKCꢀα, but also other enzymes involved into the
process of MDR induction are the intracellular target (tarꢀ
gets) of the compounds obtained, which makes them valuꢀ
able instruments for the study of molecular mechanisms of
the MDR induction in tumor cells under the action of
chemotherapeutic drugs. The practical value of the new
series of compounds consists also in the possibility of
choosing a particular derivative depending on the properꢀ
ties required. Cytotoxic compounds can serve as the protoꢀ
types of new antitumor drugs, which not only suppress
the growth of tumor cells, but also prevent the emergence
of MDR. The lowꢀtoxic compounds can be used in combiꢀ
nation with the known cytostatic drugs for prevention of
the MDR emergence during chemotherapy.
Comꢀ
pound
IC50
/nmol L–1
InC50
PC50
μmol L–1
8a*
8b*
8c**
8d**
8e**
8f**
8g*
8h*
9b*
9c*
90
—
94
125
—
—
117
135
>250
—
—
—
150
>250
—
>250
130
>250
121
—
16 4
18 5
>50
>50
>50
5.4
7.3
5.2
1.9
3.3
2.2
1.8
2.1
10
7.3
6.6
4.6
3.7
6.6
6.2
5.4
5.4
5.4
3.3
5.4
6.1
4.6
4.5
>50
20 4
10+3
12 4
12 4
16 4
16 3
12 3
19+2
14 3
18+2
17+3
7 3
5 3
>50
12 3
>50
5 1
9d*
9e*
10b*
10c*
10d*
10e*
11*
12***
15a***
15b**
15c***
15d**
BisꢀI
Experimental
Melting points were measured on a Buchi SMPꢀ20 appaꢀ
ratus. 1H and 13C NMR spectra were recorded on a Varian
VXRꢀ400 spectrometer (1H, 400 MHz; 13C, 75 MHz) using the
solvent signal as the standard. Mass spectra were obtained on a
Finnigan SAQ 710 instrument (70 eV, direct inlet, temperature
of the ions source, 150 °C) using the electron impact technique
(EIꢀMS). Analytical HPLC was performed on a Shimadzu LC10
chromatograph on a Gemini C18 column (4.6×250 mm) with the
particle size of 5 μm (Phenomenex, USA). Detection was perꢀ
formed on a Shimadzu UV—Vis 10A spectrophotometer at the
wavelength of 254 nm. The mobile phase was composed of 0.2%
of HCOONH4 (A) and acetonitrile (В). The elution was carried
out in a gradient regime during which the percentage of acetoniꢀ
trile (В) increased from 30 to 90% in 20 min and kept at 90% for
10 min at a flow rate of 1 mL min–1. The volume of the injector
loop was 10 μm, the samples were injected up to concentrations
of 0.01—0.05 mg mL–1 in aqueous acetonitrile (1 : 1). Analytical
TLC was performed on Silica gel F254 plates (Merck), column
chromatography, on Silica gel Merck 60. Extracts were dried
with anhydrous Na2SO4, solvents were evaporated under reduced
pressure. Reagents and solvents, unless otherwise stated, were
obtained from commercial sources. Data from HPLC, mass specꢀ
trometry, and NMR spectroscopy, as well as melting points of
crystalline products are given in Tables 2 and 3.
—
—
102
Note. IC50 is the concentration of compound inꢀ
ducing a decrease in activity of PKCꢀα by 50%
(in a number of cases, (–) IC50 was not determined);
InC50 is the concentration of compound inducing
apoptosis of 50% of cells (in all the experiments, the
corresponding values in the samples under monitorꢀ
ing without addition of compounds under study were
taken as 100%); PC50 is the concentration of
compound decreasing activation of gene MDR1
with Cytosar by 50% (the cells treated only
with Cytosar were taken as having 100% activation
of MDR1).
* Moderately toxic, ** low toxic, *** highly toxic
compounds (for explanation, see text).
stitution at the maleimide nitrogen atom leads to a sharp
decrease in activity. This allows us to suggest that the inꢀ
teraction of new inhibitors with the active site of the enꢀ
zyme can differ from the binding in the bis(indolꢀ3ꢀ
yl)maleimide derivatives since it is known that the NH
group of the maleimide fragment is involved in the formaꢀ
tion of one of important hydrogen bonds between the inꢀ
hibitor and the enzyme.1,8
It is of note that the data given in Table 1 suggest the
absence of direct correlation between the inhibitory activiꢀ
ty with respect to PKCꢀα in vitro and suppression of the
MDR induction in cells under the action of Cytosar. Thus
the values of IC50 for 8c and 8d are virtually the same,
whereas the values of PC50 differ by a factor of about 3.
1ꢀ(3ꢀDimethylaminopropyl)ꢀ3ꢀ(2,3ꢀdihydroindolꢀ1ꢀyl)ꢀ4ꢀ(inꢀ
dolꢀ1ꢀyl)maleimide (8h). Excess of 3ꢀ(dimethylamino)propyl
chloride hydrochloride and К2CO3 (5 equiv.) were added to
a solution of 3ꢀ(2,3ꢀdihydroindolꢀ1ꢀyl)ꢀ4ꢀ(indolꢀ1ꢀyl)maleimide
(1 g) in dioxane (30 mL), the mixture was refluxed with stirring
for ~16 h and filtered, the filtrate was concentrated. The residue
was dissolved in ethyl acetate (100 mL), the solution was washed
with brine (50 mL) and dried, the solvent was evaporated. After
chromatography (CHCl3—MeOH—Et3N, 6 : 1 : 0.1), compound
8h was obtained as a red solid; the yield was 0.86 g (68%).
Aminomethylation of compounds 1—4, 6, and 7 (general proꢀ
cedure). Paraformaldehyde (500 mg) and the corresponding
amine (8 mmol) were added to a solution of the starting indolylꢀ
maleimide (2 mmol) in acetic acid (50 mL). For the preparation
of dimethylamino derivatives, 40% aq. Me2NH was used. The

3ꢀ(Dialkylaminomethylindolꢀ1ꢀyl)maleimides
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
2015
Table 2. Analytical properties of compounds obtained
Comꢀ
poꢀ
und
Molecular
formula
(Мw)
M.p./°С
HPLC data
Content
Comꢀ
poꢀ
und
Molecular
formula
(Мw)
M.p./°С
HPLC data
Content
Rt
Rt
/min
(%)
/min
(%)
8a
8b
8c
8d
8e
8f
C25H26N4O2
(414.5)
253—255
11.05
98.6
10c
10d
10e
11
C26H28N4O2
(428.53)
Amorphous 16.70
Amorphous 13.61
98.1
C25H24N4O2
(412.48)
Amorphous 10.75
Amorphous 12.33
97.6
97.1
98.0
97.8
98.8
97.9
98.3
97.9
99.0
98.5
98.6
98.8
98.0
97.4
C26H28N4O3
(444.53)
98.2
97.9
98.1
98.5
97.7
97.0
97.6
97.8
98.7
97.9
98.3
98.2
97.6
99.0
C24H24N4O2
(400.47)
C27H31N5O2
(457.57)
125—126
Amorphous 14.95
115—117 14.68
12.87
C26H26N4O2
(426.51)
145—146
12.98
16.64
C28H32N4O2
(456.58)
C26H26N4O3
(442.51)
205—207
(fumarate)
12
C26H27ClN4O2
(462.97)
C27H29N5O2
(455.55)
Amorphous 12.56
Amorphous 14.28
13
C29H32N4O4
(500.59)
Amorphous 13.29
Amorphous 13.04
8g
8h
9a
9b
9c
9d
9e
10a
10b
C27H30N4O2
(442.55)
14a
14b
14c
14d
14e
15a
15b
15c
15d
C24H22N4O2
(398.46)
C30H37N5O2
(499.65)
Amorphous
8.58
C26H26N4O2
(426.51)
107—108
(fumarate)
14.07
13.65
18.64
12.57
3.65
C23H23N3O3
(421.51)
Amorphous 12.81
C26H24N4O2
(424.49)
95—97
(fumarate)
C25H27N3O3
(449.57)
199—200
(fumarate)
8.4
C26H24N4O3
(440.49)
204—205
C25H25N3O3
(447.55)
212—213
(fumarate)
18.33
13.52
12.77
C27H27N5O2
(453.54)
110—111
(fumarate)
C25H25N3O4
(463.55)
118—119
C27H29N5O2
(455.55)
Amorphous
C26H28N4O3
476.59
228—230
(fumarate)
C31H33N5O2
(507.63)
Amorphous
3.47
C24H26N4O2
(402.49)
Amorphous 12.66
C31H33N5O4
(539.62)
Amorphous 15.24
Amorphous 8.11
C26H30N4O2
(430.54)
95—96 13.77
C33H39N7O2
(565.71)
reaction mixture was stirred for 20 h at 50 °C and concentrated,
the residue was dissolved in ethyl acetate (100 mL), the solution
was washed with saturated aq. NaHCO3 (2×30 mL) and water
and dried, the solvent was evaporated. The residue was subjected
to chromatography (EtOAc—PriOH—25% aq. NH3, 24 : 8 : 1)
to isolate the corresponding dialkylaminomethyl derivatives as
red solids in 50—75% yield.
Aminomethylation of Nꢀmethylꢀ3,4ꢀbis(indolꢀ1ꢀyl)maleimide
(5). Paraformaldehyde (500 mg) and the corresponding amine
(25 mmol) were added to a solution of compound 5 (1 g,
2.9 mmol) in acetic acid (50 mL). Under conditions described
above, the corresponding monoꢀ and bis(dialkylaminomethyl)
derivatives were obtained as red solids in 50 and 20% yields,
respectively.
(20 mL), and dried in vacuo to obtain fumarates of the correꢀ
sponding compounds (1—4 and 6—9) as orange crystalline powꢀ
ders in 50—70% yield.
Inhibition of activity of PKCꢀα by new bis(indolyl)maleimide
derivatives. The ativity of the human recombinant PKCꢀα (Calꢀ
biochem, USA) was assayed based on the incorporation of 32P
from ATP into the total histone from calf tymus* in the optiꢀ
mized cellꢀfree test system.
The reaction was carried out in a buffer solution of the folꢀ
lowing composition: trisꢀHCl (20 mmol L–1), MgCl2 (5 mmol L–1),
CaCl2 (1 mmol L–1), NaCl (200 mmol L–1), 2ꢀmercaptoethanol
(6 mmol L–1), 0.01% Triton Xꢀ100, pH 7.5 (at 30 °C). The reacꢀ
tion mixture was prepared from the buffer solution (0.1 mL) by
addition of the inhibitor under study dissolved in DMSO to the
Synthesis of fumarates of aminomethyl derivatives. Equimolar
amount of fumaric acid (saturated solution in anhydrous diethyl
ether) was added to a solution of compounds 1—4 and 6—9
(200—300 mg) in anhydrous diethyl ether (100 mL). The precipꢀ
itate that formed was filtered off, washed with anhydrous ether
final concentrations in a mixture of 50, 100, and 250 nmol L–1
.
* Generous gift from the Academician L. L. Kiselev laboratory at
the V. A. Engel'hardt Institute of Molecular Biology of the Rusꢀ
sian Academy of Sciences.

2016
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
Simonov et al.
Table 3. Spectral data of compounds obtained
Comꢀ
pound
MS,
m/z (Irel (%))
NMR, δ (J/Hz)
1H
13C
8a
8b
8c
8d
8e
8f
414 [М]+ (25),
342 [М – NEt2]+
(100)
0.97—1.01 (m, 6 H); 2.40—2.46 (m, 4 H); 3.08 (t, 2 H,
J = 8.1); 3.66 (s, 2 H); 4.27 (t, 2 H, J = 8.1); 6.01
(d, 1 H); 6.41 (t, 1 H, J = 7.1); 6.61 (t, 1 H, J = 7.4);
6.93, 6.99 (both t, 1 H each, J = 6.9); 7.02
(d, 1 H, J = 8.3); 7.14 (d, 1 H, J = 7.9); 7.24
(s, 1 H); 7.55 (d, 1 H, J = 7.2); 10.8 (s, 1 H)
1.83 (t, 4 H, J = 2.1); 2.90 (t, 4 H, J = 4.0); 3.09
(t, 2 H, J = 7.1); 4.23 (s, 2 H); 4.29 (t, 2 H, J = 8.0);
6.02 (d, 1 H, J = 8.1); 6.47 (t, 1 H, J = 7.5); 6.63
(t, 1 H, J = 7.4); 6.99—7.07 (m, 3 H); 7.21 (d, 1 H,
J = 7.5); 7.51 (s, 1 H); 7.64 (d, 1 H, J = 7.2);
10.5 (s, 1 H)
2.13 (s, 6 H); 3.01 (s, 3 H); 3.10 (t, 2 H, J = 8.0);
3.52 (s, 4 H); 4.29 (t, 2 H, J = 7.9); 6.05 (d, 1 H,
J = 8.1); 6.44, 6.62 (both t, 1 H each, J = 7.4);
6.93 (t, 1 H, J = 7.5); 6.99 (t, 1 H, J = 7.9);
7.03 (d, 122.3, 124.3, 126.0, 127.2, 1 H, J = 7.0);
7.14 (d, 1 H, J = 8.0); 7.26 (s, 1 H); 7.52 (d, 1 H, J = 7.7)
1.66, 2.39 (both s, 4 H each); 3.01 (s, 3 H); 3.09 (t, 2 H,
J = 8.0); 3.69 (s, 2 H); 4.27 (t, 2 H, J = 8.1); 6.02
(d, 1 H, J = 8.1); 6.42 (t, 1 H, J = 7.6); 6.63 (t, 1 H,
J = 7.3); 6.94 (t, 1 H, J = 7.5); 6.99 (t, 1 H, J = 7.0);
7.03 (d, 1 H, J = 7.3); 7.14 (d, 1 H, J = 7.7); 7.24
(s, 1 H); 7.52 (d, 1 H, J = 7.3)
11.7 (2 C), 28.8, 45.8 (2 C),
47.6, 52.3, 108.9, 110.8,
111.6, 114.9, 119.3, 119.7,
121.9, 122.0, 124.2, 125.8,
127.1, 127.5, 131.7, 132.3,
136.8, 142.6, 167.5, 168.3
22.5 (2 C), 28.8, 47.2, 51.7
(2 C), 52.5, 107.9, 111.2,
111.8, 118.7, 120.4, 122.3,
122.4, 124.3, 125.9, 127.0,
129.7, 132.1, 133.2, 136.5,
142.4, 167.3, 168.2
23.6, 28.8, 44.7 (2 C), 52.5,
53.9, 108.0, 110.9, 111.8,
119.8, 114.8, 119.4, 122.0,
122.3, 124.3, 126.0,127.2,
127.4, 131.9, 132.4, 136.9,
142.5, 166.4, 167.5
23.1 (2 C), 23.7, 28.8, 49.7,
52.5, 110.9, 53.2 (2 C), 108.1,
111.8, 115.3, 119.1, 119.8,
122.0, 122.3, 124.3, 125.9,
126.8, 127.3, 131.9, 132.4,
136.8, 142.5, 166.4, 167.5
23.7, 28.8, 52.5, 52.8 (2 C),
52.9, 66.3 (2 C), 107.9, 111.0,
111.8, 113.4, 119.3, 120.0,
122.1, 122.4, 124.3, 125.9,
127.5 (2 C), 132.0, 132.6,
136.8, 142.5, 166.5, 167.5
23.7, 28.9, 43.7, 50.3 (2 C),
51.8, 52.6, 53.1 (2 C), 107.7,
111.1, 111.8, 112.7, 119.3,
120.1, 122.2, 122.4, 124.4,
125.9, 127.4, 128.0, 132.1,
132.8, 134.7 (2 C), 136.8,
142.4, 166.4, 167.2 (2 C),
167.5 (fumarate)
412 [М]+ (40),
342 [М – N(C4H8)]+
(100)
400 [М]+ (30),
356 [М – NMe2]+
(100)
426 [М]+ (35),
356 [М – NC4H8]+
(100)
442 [М]+ (53),
2.29 (s, 4 H); 3.01 (s, 3 H); 3.10 (t, 2 H, J = 8.0); 3.54
356 [М – NC4H8O]+ (s, 4 H); 3.58 (s, 2 H); 4.30 (t, 2 H, J = 7.9); 5.96
(100)
(d, 1 H, J = 8.0); 6.39 (t, 1 H, J = 7.7); 6.62 (t, 1 H,
J = 7.3); 6.96 (t, 1 H, J = 7.7); 7.02 (t, 1 H, J = 7.5);
7.04 (d, 1 H, J = 6.9); 7.18 (d, 1 H, J = 8.1); 7.22
(s, 1 H); 7.56 (d, 1 H, J = 7.6)
2.43 (s, 3 H); 2.50—2.51, 2.73—2.74 (both m, 4 H each);
3.01 (s, 3 H); 3.10 (t, 2 H, J = 7.9); 3.69 (s, 2 H); 4.30
(t, 2 H, J = 7.9); 5.98 (d, 1 H, J = 7.9); 6.42 (t, 1 H,
J = 7.7); 6.55 (s, 2 H); 6.63 (t, 1 H, J = 7.4); 6.97
(t, 1 H, J = 7.7); 7.03 (t, 1 H, J = 8.1); 7.04 (d, 1 H,
J = 7.4); 7.19 (d, 1 H, J = 8.1); 7.27 (s, 1 H); 7.57
(d, 1 H, J = 7.5) (fumarate)
455 [М]+ (100), 356
[М – NC4H8NMe]+
(42)
8g
8h
442 [М]+ (25), 370
[М – NEt2]+
(100)
0.86—0.99 (m, 6 H); 1.18—1.21 (m, 3 H); 2.40—2.45
(m, 4 H); 3.24 (t, 2 H, J = 8.0); 3.57—3.59 (m, 2 H);
3.65 (s, 2 H); 4.30 (t, 2 H, J = 8.1); 6.00 (d, 1 H,
J = 8.0); 6.42 (t, 1 H, J = 7.7); 6.63 (t, 1 H, J = 7.5);
6.94 (t, 1 H, J = 7.1); 6.99 (t, 1 H, J = 7.0); 7.03
(d, 1 H, J = 7.3); 7.14 (d, 1 H, J = 8.0); 7.23 (s, 1 H);
7.56 (d, 1 H, J = 7.6)
0.92—1.00 (m, 12 Н); 1.71—1.74 (m, 2 Н); 2.38—2.48
(m, 10 Н); 3.09 (t, 2 Н, J = 7.9); 3.56 (t, 2 Н, J = 6.8);
3.64 (s, 2 Н); 4.30 (t, 2 Н, J = 8.0); 5.95 (d, 1 Н,
J = 8.1); 6.40 (t, 1 Н, J = 8.0); 6.61 (t, 1 Н, J = 7.3);
6.93 (t, 1 Н, J = 7.7); 6.99 (t, 1 Н, J = 8.3); 7.30
(d, 1 Н, J = 7.3); 7.12 (d, 1 Н, J = 8.1); 7.22 (s, 1 Н);
7.55 (d, 1 Н, J = 7.3)
11.7 (2 C), 13.7, 28.8, 32.3,
45.8 (2 C), 47.6, 52.4, 107.9,
110.9, 111.7, 115.2, 119.3,
119.7, 121.9, 122.2, 124.2,
125.8, 127.0, 127.52, 131.8,
132.0, 136.9, 142.5, 166.1,
167.1
11.5 (2 C), 11.8 (2 C), 25.6,
28.8, 36.1, 45.9 (2 C), 46.1
(2 C), 47.5, 49.9, 52.5, 107.8,
110.9, 111.6, 115.2, 119.4,
119.7, 122.00, 122.3, 124.3,
125.9, 127.0, 127.5, 131.8,
131.9, 136.9, 142.5, 166.3,
167.4
413 [М –
(CH2)3NMe2]+
(40), 342 [М –
(CH2)3NMe2,
NEt2]+ (100)
(to be continued)

3ꢀ(Dialkylaminomethylindolꢀ1ꢀyl)maleimides
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
2017
Table 3 (continued)
Comꢀ
pound
MS,
m/z (Irel (%))
NMR, δ (J/Hz)
1H
13C
9a
9b
421 [М]+ (56),
377 [М – NMe2]+
(100)
2.11 (s, 6 H); 3.02 (s, 3 H); 3.39 (s, 2 H); 3.54 (s, 3 H);
6.38, 6.96 (both d, 2 H each, J = 8.8); 7.07 (s, 1 H);
7.07 (t, 1 H, J = 6.5); 7.19 (t, 1 H, J = 7.1);
24.2, 44.9 (2 C), 53.8, 54.9,
112.5, 113.8 (2 C), 116.2,
117.4, 119.3, 120.8, 122.2,
126.1, 126.3, 128.2, 132.1
(2 C), 132.4, 135.2, 159.0,
166.4, 167.6
7.29 (d, 1 H, J = 8.2); 7.49 (d, 1 H, J = 7.7)
449 [М]+ (35),
377 [М – NEt2]+
(100)
1.10—1.16 (m, 6 H); 2.69—2.71 (m, 4 H); 3.02, 3.55
(both s, 3 H each); 3.93 (s, 2 H); 6.39 (d, 2 H, J = 8.9);
6.57 (s, 2 Н); 6.98 (d, 2 H, J = 8.8); 7.11 (t, 1 H,
J = 7.1); 7.22 (t, 1 H, J = 7.2); 7.29 (s, 1 H); 7.33
(d, 1 H, J = 8.2); 7.57 (d, 1 H, J = 7.9) (fumarate)
10.1 (2 C), 24.1, 45.1 (2 C),
46.1, 55.0, 112.6, 113.8 (2 C),
117.0, 118.9, 121.0, 122.5,
128.0, 128.0, 128.2, 131.8,
132.3 (2 C), 134.6 (2 C),
135.2, 159.1, 166.3, 167.1
(2 C), 167.3 (fumarate)
9c
47 [М]+ (15), 377
[М – NC4H8]+
(100)
1.83, 2.84 (both s, 4 H each); 3.02, 3.55 (both s, 3 H each);
4.02 (s, 2 H); 6.39 (d, 2 H, J = 8.9); 6.55 (s, 2 H);
6.97 (t, 2 H, J = 8.9); 7.11 (t, 1 H, J = 6.9); 7.22
(t, 1 H, J = 7.1); 7.29 (s, 1 H); 7.31 (d, 1 H, J = 8.2);
7.58 (d, 1 H, J = 7.9) (fumarate)
22.7 (2 C), 24.1, 47.7, 52.3
(2 C), 55.0, 112.5, 113.8 (2 C),
116,9, 118.9, 118.8, 121.1,
122.5, 127.7, 128.2, 134.7
(2 C), 128.3, 131.6, 132.3
(2 C), 135.1, 159.1 (2 C),
166.3, 167.3 (2 C) (fumarate)
9d
463 [М]+ (37),
2.32 (s, 4 H); 3.02 (s, 3 H); 3.48 (s, 2 H); 3.54 (s, 3 H);
24.2, 51.8, 53.1 (2 C), 55.0,
66.2 (2 C), 112.5, 113.8 (2 C),
115.1, 117.3, 119.3, 120.8,
122.3, 126.3 (2 C), 126.5,
128.2, 132.1 (2 C), 132.2,
135.3, 159.0, 166.5, 167.6
377 [М – NC4H8O]+ 3.57 (s, 2 H); 6.37, 6.96 (both d, 2 H each, J = 9.0);
(100)
7.08 (t, 1 H, J = 7.0); 7.09 (s, 1 H); 7.19 (t, 1 H,
J = 7.1); 7.29 (d, 1 H, J = 8.1); 7.54 (d, 1 H, J = 7.8)
9e
476 [М]+ (20), 377
[М – NC4H8NMe]+
(100)
2.27 (s, 3 H); 2.43—2.51 (m, 8 H); 3.02 (s, 3 H); 3.51
(s, 2 H); 3.56 (c, 3 H); 6.38 (d, 2 H, J = 8.9); 6.97
(t, 2 H, J = 8.8); 7.07 (t, 1 H, J = 7.8); 7.09 (s, 1 H);
7.19 (t, 1 H, J = 7.1); 7.29 (d, 1 H, J = 8.3);
7.53 (d, 1 H, J = 7.7)
24.1, 44.8, 51.6 (2 C), 52.4,
54.1 (2 C), 55.0, 112.4, 113.7
(2 C), 115.2, 117.3, 119.2,
120.7, 122.2, 126.3, 126.7,
128.1, 132.1 (2 C), 132.2,
135.3, 159.0, 166.3, 167.4
10a
402 [М]+ (25), 358
[М – NMe2]+
(100)
1.05 (m, 3 H); 2.46 (s, 6 H); 2.98 (s, 3 H); 4.00—4.06
(m, 4 H); 6.56 (s, 2 H); 6.74 (t, 1 H, J = 7.3); 6.85
(t, 2 H, J = 8.2); 6.98 (d, 2 H, J = 8.7); 7.03
(t, 1 H, J = 7.8); 7.12 (t, 1 H, J = 7.1); 7.19 (s, 1 H);
7.23 (d, 1 H, J = 8.2); 7.55 (d, 1 H, J = 7.8) (fumarate)
13.8, 23.7, 41.5 (2 C), 46.5,
50.4, 105.8, 107.2, 111.5,
118.5, 120.4, 122.1, 123.0
(2 C), 124.8, 127.5, 127.9
(2 C), 130.7, 134.8 (2 C),
136.3, 138.5, 141.4, 166.0,
167.5, 167.6 (2C) (fumarate)
10b
10c
430 [М]+ (25), 358
[М – NEt2]+
(100)
0.98 (t, 6 H, J = 7.1); 1.04 (t, 3 H, J = 0.9); 2.39
(m, 4 H); 2.97 (s, 3 H); 3.52 (s, 2 H); 3.89 (m, 2 H);
6.78 (t, 1 H, J = 7.3); 6.88 (t, 2 H, J = 8.1); 6.94
(s, 1 H); 6.96—7.01 (m, 3 H); 7.07 (t, 1 H, J = 7.1);
7.17 (t, 1 H, J = 8.1); 7.49 (d, 1 H, J = 7.8)
11.8 (2 C), 13.7, 23.6, 45.9
(2 C), 46.1, 47.8, 108.0, 111.1,
114.4, 119.2, 119.5, 121.7,
122.3 (2 C), 124.2, 127.1,
127.8, 127.8 (2 C), 136.6,
137.3, 141.8, 166.2, 167.6
428 [М]+ (21), 358
[М – NC4H8]+
(100)
1.04 (t, 3 H, J = 6.3); 1.67, 2.37 (both s, 4 H each);
2.97 (s, 3 H); 3.58 (s, 2 H); 3.89—3.92 (m, 2 H); 6.78
(t, 1 H, J = 7.4); 6.88 (t, 2 H, J = 7.5); 6.95 (s, 1 H);
6.97—7.00 (m, 3 H); 7.07 (t, 1 H, J = 7.1); 7.17 (d, 1 H,
J = 8.2); 7.47 (d, 1 H, J = 7.8)
13.7, 23.1 (2 C), 23.6, 46.2,
49.9, 53.4 (2 C), 107.9, 111.1,
114.6, 119.0, 119.6, 121.7,
122.4 (2 C), 124.3, 127.0,
127.6, 127.8 (2 C), 136.5,
137.4, 141.8, 166.2, 167.6
(to be continued)

2018
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
Simonov et al.
Table 3 (continued)
Comꢀ
pound
MS,
m/z (Irel (%))
NMR, δ (J/Hz)
1H
13C
10d
10e
444 [М]+ (30), 358
[М – NC4H8O]+
(100)
1.10 (t, 3 H, J = 6.9); 2.29 (s, 4 H); 3.00 (s, 3 H);
3.48 (s, 2 H); 3.58 (s, 4 H); 3.97 (m, 2 H); 6.78 (t, 1 H,
J = 7.4); 6.74 (t, 1 H, J = 7.3); 6.84 (t, 2 H, J = 7.5);
6.94 (s, 1 H); 6.97—7.02 (m, 3 H); 7.10 (t, 1 H,
J = 7.9); 7.53 (d, 1 H, J = 7.8)
13.7, 23.5, 46.2, 53.0 (2 C),
53.1, 66.3 (2 C), 107.5, 111.1,
112.8, 119.1, 119.6, 121.7,
122.4 (2 C), 124.3, 127.5,
127.6, 127.8 (2 C), 136.5,
137.4, 141.5, 166.2, 167.4
13.6, 23.5, 45.6, 46.1, 52.3
(2 C), 52.7, 54.7 (2 C), 107.5,
111.0, 113.2, 119.0, 119.5,
121.6, 122.4 (2 C), 124.2,
127.3, 127.6, 127.7 (2 C),
136.5, 137.5, 141.6, 166.1,
167.4
457 [М]+ (20), 358
[М – NC4H8NMe]+
(100)
1.06 (t, 3 H, J = 6.3); 2.16 (s, 3 H); 2.31 (s, 4 H);
2.97 (s, 3 H); 3.45 (s, 6 H); 3.94 (m, 2 H); 6.75 (t, 1 H,
J = 7.3); 6.85 (t, 2 H, J = 8.2); 6.91 (s, 1 H);
6.96—6.99 (m, 3 H); 7.07 (t, 1 H, J = 7.1); 7.17
(d, 1 H, J = 8.1); 7.48 (d, 1 H, J = 7.8)
11
456 [М]+ (37), 384
[М – NEt2]+
(100)
0.96—1.02 (m, 6 Н); 1.93 (t, 2 Н, J = 5.7); 1.98
(s, 3 Н); 2.38—2.43 (m, 4 Н); 2.63 (t, 2 Н, J = 6.6);
3.00 (s, 3 Н); 3.59 (s, 2 Н); 3.72 (t, 2 Н, J = 5.3);
6.23 (d, 1 Н, J = 8.3); 6.49 (d, 1 Н, J = 8.2);
6.60 (s, 1 Н); 6.28 (t, 1 Н, J = 7.2); 7.00 (s, 1 Н);
7.05 (t, 1 Н, J = 7.3); 7.18 (d, 1 Н, J = 8.1);
7.54 (d, 1 Н, J = 7.7)
11.7 (2 C), 20.0, 22.4, 23.5,
25.5, 45.7 (2 C), 47.4, 47.9,
111.2, 111.4, 114.8, 119.0,
119.1, 119.7, 121.6, 125.4,
126.4, 126.9, 128.0, 128.6,
131.1, 134.5, 135.2, 136.0,
166.7, 167.2
12
13
C464 [М]+ (10), 462
[М]+ (25), 391 [М –
NEt2]+ (40),
1.01—1.04 (m, 6 Н); 2.46—2.51 (m, 4 Н); 3.02 (s, 3 Н);
3.11 (t, 2 Н, J = 8.1); 3.44 (s, 2 Н); 4.31 (t, 2 Н,
J = 7.1); 5.95, 6.41 (both d, 1 Н each, J = 8.6);
6.99 (t, 1 Н, J = 7.1); 7.04 (t, 1 Н, J = 7.9); 7.07
(d, 1 Н, J = 7.2); 7.17 (d, 1 Н, J = 7.8); 7.25 (s, 1 Н);
7.61 (d, 1 Н, J = 7.7)
0.95—1.03 (m, 6 Н); 2.38—2.44 (m, 4 Н); 2.68—2.87
(m, 2 Н); 3.01 (s, 3 Н); 3.66 (s, 2 Н); 3.67 (s, 3 Н);
3.67—3.73 (m, 2 Н); 4.55—4.60 (m, 1 Н); 5.87 (d, 1 Н,
J = 8.1); 6.41 (t, 1 Н, J = 7.8); 6.64 (t, 1 Н, J = 7.3);
6.96 (t, 1 Н, J = 7.2); 7.00 (t, 1 Н, J = 6.6); 7.06
(d, 1 Н, J = 7.5); 7.20 (d, 1 Н, J = 7.1); 7.21 (s, 1 Н);
7.57 (d, 1 Н, J = 8.1)
11.4 (2 C), 23.4, 28.5, 45.8
(2 C), 47.4, 52.4, 109.1, 110.8,
112.4, 119.3, 119.8, 122.0,
124.1, 125.4, 125.7, 127.0
(2 C), 127.4, 131.7, 134.1,
136.5, 141.5, 166.1, 167.0
11.7 (2 C), 23.5, 37.0, 37.7,
45.8 (2 C), 47.5, 51.4, 58.0,
108.0, 111.1, 111.6, 115.2,
119.4, 119.8, 121.9, 122.3,
123.6, 126.5, 126.9, 127.5,
131.9, 133.8, 136.7, 142.2,
166.5, 167.4, 171.8
390 [М – NEt2]+
(100)
500 [М]+ (60), 428
[М – NEt2]+
(100)
14a
14b
398 [М]+ (53), 354
[М – NMe2]+
(100)
2.16 (s, 6 H); 3.12 (s, 3 H); 3.55 (s, 2 H); 6.57 (t, 2 H,
J = 9.4); 6.67 (t, 1 H, J = 7.2); 6.73 (d, 1 H, J = 7.3);
6.79 (t, 1 H, J = 3.4); 6.90 (d, 2 H, J = 5.5); 7.46
(d, 1 H, J = 10.1); 7.47 (s, 1 H); 7.52 (d, 1 H, J = 7.8);
7.65 (d, 1 H, J = 3.5)
24.1, 44.7 (2 C), 106.6, 110.7,
110.8, 117.3, 119.7, 120.6,
121.1, 121.4, 122.3, 122.6
(2 C), 122.6, 123.0, 126.1,
127.8, 128.2, 128.32, 134.9,
135.4, 166.6, 166.6
11.7 (2 C), 24.0, 46.0 (2 C),
47.5, 108.5, 110.7, 110.8,
117.9, 119.6, 120.5, 121.0,
121.3, 122.1, 122.5 (2 С),
123.1, 125.8, 127.7, 128.2,
128.3, 134.9, 135.4, 166.5,
166.5
426 [М]+ (20), 354
[М – NEt2]+
(100), 339 [М –
NEt2 – Me]+
(12)
1.00 (m, 6 H); 2.46 (m, 4 H); 3.12 (s, 3 H); 3.69 (s, 2 H);
6.55 (d, 1 H, J = 8.4); 6.65 (d, 1 H, J = 6.2); 6.67
(t, 1 H, J = 7.7); 6.75 (d, 1 H, J = 8.3); 6.78 (d, 1 H,
J = 4.1); 6.91 (t, 1 H, J = 4.0); 6.93 (t, 1 H, J = 4.0);
7.44 (s, 1 H); 7.47 (d, 1 H, J = 7.8); 7.56 (d, 1 H,
J = 7.7); 7.64 (d, 1 H, J = 3.4)
14c
424 [М]+ (22),
354 [М – NC4H8]+
(100)
1.69, 2.44 (both s, 4 H each); 3.10 (s, 3 H); 3.75 (s, 2 H);
6.50 (d, 1 H, J = 7.6); 6.59 (d, 1 H, J = 8.4); 6.62
(t, 1 H, J = 7.2); 6.72 (t, 1 H, J = 8.3); 6.74 (d, 1 H,
J = 3.5); 6.88 (t, 1 H, J = 7.0); 6.91 (t, 1 H, J = 7.9);
7.42 (d, 1 H, J = 6.9); 7.43 (s, 1 H); 7.51 (d, 1 H,
J = 7.8); 7.60 (d, 1 H, J = 3.5)
23.1 (2 C), 23.7, 52.5, 53.2
(2 C), 108.1, 110.9, 111.8,
115.3, 119.1, 119.8, 122.0,
122.3, 124.3, 125.9, 126.8
(2 C), 127.3 (2 C), 131.95,
132.4, 136.8, 142.5, 166.4,
167.5
(to be continued)

3ꢀ(Dialkylaminomethylindolꢀ1ꢀyl)maleimides
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
2019
Table 3 (continued)
Comꢀ
pound
MS,
m/z (Irel (%))
NMR, δ (J/Hz)
1H
13C
14d
14e
440 [М]+ (25), 354
[М – NC4H8O]+
(100)
2.31—2.35 (m, 4 H); 3.12 (s, 3 H); 3.44—3.85 (m, 4 H);
3.62 (s, 2 H); 6.52 (d, 1 H, J = 7.6); 6.64 (t, 1 H,
J = 7.0); 6.71 (d, 1 H, J = 8.2); 6.78 (t, 1 H, J = 3.3);
6.80 (d, 1 H, J = 5.5); 6.91 (t, 1 H, J = 8.1); 6.95
(t, 1 H, J = 7.1); 7.39 (s, 1 H); 7.46 (d, 1 H, J = 7.8);
7.58 (d, 1 H, J = 7.9); 7.64 (d, 1 H, J = 3.5)
24.1, 51.5 (2 C), 52.8, 66.1
(2 C), 106.6, 110.7, 111.0,
116.1, 119.6, 120.7, 121.2,
121.4, 122.3, 122.6, 122.9,
123.1, 126.3, 127.8, 128.3,
128.3, 134.9, 135.4, 166.5,
166.2
458 [М]+ (100),
354 [М –
NC4H8NMe]+ (47)
2.16 (s, 3 H); 2.22—2.40 (m, 8 H); 3.12 (s, 3 H);
3.61 (s, 2 H); 6.53 (d, 1 H, J = 8.2); 6.65 (d, 1 H,
J = 7.4); 6.69 (d, 1 H, J = 7.9); 6.77 (t, 1 H,
J = 7.4); 6.78 (t, 1 H, J = 3.8); 6.91 (t, 1 H,
J = 7.5); 6.94 (t, 1 H, J = 7.7); 7.39 (s, 1 H);
7.46, 7.55 (both d, 1 H each, J = 7.7);
7.64 (d, 1 H, J = 3.4)
24.0, 45.6, 52.2 (2 C), 52.4,
54.7 (2 C), 106.5, 110.7,
110.9, 126.7, 119.6, 120.5,
121.1, 121.3, 122.2, 122.5,
122.7, 123.0, 126.0, 127.7,
128.2, 128.3, 134.9, 135.4,
166.4, 166.5
15a
15b
15c
15d
455 [М]+ (25), 411
[М – NMe2]+
(100), 368 [М –
2 NMe2]+ (50)
2.16 (s, 12 H); 3.11 (s, 3 H); 3.56 (s, 4 H); 6.56 (d, 2 H,
J = 8.2); 6.67 (t, 2 H, J = 8.2); 6.89 (t, 2 H, J = 7.9);
7.49 (s, 2 H); 7.51 (d, 2 H, J = 7.9)
24.1, 44.7 (4 C), 53.6 (2 C),
110.7 (2 C), 117.2 (2 C),
119.7 (2 C), 121.1 (2 C),
122.4 (2 C), 122.7 (2 C),
126.1 (2 C), 128.2 (2 C),
135.4 (2 C), 166.6 (2 C)
507 [М]+ (30), 436
[М – NC4H8]+ (50),
367 [М – 2 NC4H8O]+ (t, 2 H, J = 6.9); 7.45 (s, 2 H); 7.52 (d, 2 H, J = 8.0)
1.69, 2.43 (both s, 8 H each); 3.10 (s, 3 H); 3.73 (s, 4 H);
6.60 (d, 2 H, J = 8.3); 6.69 (d, 2 H, J = 7.7); 6.92
23.1 (4 C), 24.0, 49.6 (2 C),
53.2 (4 C), 110.9 (2 C),
117.8 (2 C), 119.4 (2 C), 121.1
(2 C), 122.4 (2 C), 122.8 (2 C),
125.7 (2 C), 128.2 (2 C),
(100)
135.3 (2 C), 166.6 (2 C)
539 [М]+ (23), 453
2.34 (s, 8 H); 3.11 (s, 3 H); 3.56 (s, 8 H); 3.63 (s, 4 H);
24.0, 52.7 (2 C), 52.8 (4 C),
66.2 (4 C), 119.6 (2 C), 110.9
(2 C), 116.0 (2 C), 121.2
(2 C), 122.5 (2 C), 123.0 (2 C),
126.7 (2 C), 128.3 (2 C),
135.3 (2 C), 116.9 (2 C)
[М – NC4H8O]+ (33), 6.65 (d, 2 H, J = 8.2); 6.70 (d, 2 H, J = 7.8); 6.93
368 [М – 2 NC4H8O]+ (t, 2 H, J = 7.4); 7.42 (s, 2 H); 7.56 (d, 2 H, J = 7.8)
(100)
565 [М]+ (8), 466
[М – NC4H8NMe]+
(100), 368 [М –
2 (NC4H8NMe)]+
(55)
2.46 (s, 6 H); 2.54, 2.77 (both s, 8 H each); 3.11 (s, 3 H);
3.72 (s, 4 H); 6.56 (s, 2 H); 6.62 (d, 2 H, J = 8.2);
6.72 (t, 2 H, J = 7.8); 6.94 (t, 2 H, J = 7.3); 7.47
(s, 2 H); 7.55 (d, 2 H, J = 7.8) (fumarate)
24.0, 43.5 (2 C), 50.3 (4 C),
51.7 (2 C), 53.1 (4 C), 110.9
(2 C), 115.7 (2 C), 119.6 (2 C),
121.3 (2 C), 122.5 (2 C), 122.9
(2 C), 126.6 (2 C), 128.2 (2 C),
134.6 (2 C), 135.3 (2 C), 166.4
(2 C), 167.3 (2 C) (fumarate)
1 min after addition of the inhibitor, [γꢀ32P]ATP (with specific
activity of 500 imp L min–1 mol–1, Phosphor Production Associꢀ
ation, RF), histones from calf tymus (50 μg), phosphatidylserine
(2.0 μg), 1,2ꢀdioleine (0.25 μg) were added to the reaction mixꢀ
ture (phosphatidylserine and diacylglycerol were suspended in
TrisꢀHCl (20 M, pH 7.5), sonicated in a UZDNꢀ1, Uꢀ42 ultraꢀ
sonic disintegrator for 1 min at a frequency of 15 kHz, and the
micellar solution was added to the reaction mixture). The reacꢀ
tion was triggered by addition of enzyme PKC (0.05 μg, 2040
Units (mg of protein)–1). The background activity of PKCꢀα was
determined in the mixture described above in the presence of
ethylene glycol tetraacetate (0.5 M) instead of Ca2+. After 4 min
of incubation at 30 °C, the reaction was stopped by addition of
cold 10% trichloroacetic acid (TCA) (1.5 mL). The acidꢀinsoluble
material was collected on Whatman GF/A glass fiber filters,
washed with cold TCA, and the radioactivity was measured using
a LS 6500 Cherenkov liquid scintillation counter (Beckman
Coulter, USA). The final concentration of DMSO in the reaction
mixture did not exceed 0.5%. The maximum consumption of ATP
in the reaction was less than 5%. The IC50 values were determiꢀ
ned graphically from the relationship of reciprocal values of PKCꢀα
activity and concentration of the inhibitor under study (Fig. 1).
The difference between the starting and the background incorpoꢀ
ration levels of 32P into histones was taken as the 100% of activity.
Investigation of cytotoxicity. The cells of the К562 and
HCТ116 lines were cultured in the Dulbecco's modified Eagle's

2020
Russ.Chem.Bull., Int.Ed., Vol. 57, No. 9, September, 2008
Simonov et al.
a–1
References
0.045
0.040
0.035
0.025
0.020
0.015
0.010
0.005
0
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IC50
150
C/nmol L–1
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This work was financially supported by the Russian
Foundation for Basic Research (Project No. 06ꢀ03ꢀ32233)
and the European Community in the framework of
the "Protein Kinases, New Drug Targets for PostꢀGeꢀ
nomic Era" Program (Contract No. LSHBꢀCTꢀ2004ꢀ
503467).
Received October 19, 2007;
in revised form July 22, 2008