Design and development of macrocyclization methods for compounds with potential tuberculocidal activity to decrease CYP450 liver cytochrome inhibition

Article abstract of DOI:10.1134/S1070363217040107

A procedure for macrocyclization of compounds with potential tuberculocidal activity was developed in order to obtaining compounds with a lower degree of inhibition of the key CYP 3A4 cytochrome.

Full text of DOI:10.1134/S1070363217040107

ISSN 1070-3632, Russian Journal of General Chemistry, 2017, Vol. 87, No. 4, pp. 717–730. © Pleiades Publishing, Ltd., 2017.
Original Russian Text © N.I. Vasilevich, E.A. Aksenova, A.A. Aksenova, I.I. Afanasyev, 2017, published in Zhurnal Obshchei Khimii, 2017, Vol. 87, No. 4,
pp. 584–596.
Design and Development of Macrocyclization Methods
for Compounds with Potential Tuberculocidal Activity
to Decrease CYP450 Liver Cytochrome Inhibition
N. I. Vasilevich*, E. A. Aksenova, A. A. Aksenova, and I. I. Afanasyev
New Scientific Technologies, LLC, ul. Geroev Panfilovtsev 20, Moscow, 125480 Russia
*e-mail: nvasilevich@asinex.com
Received October 20, 2016
Abstract—A procedure for macrocyclization of compounds with potential tuberculocidal activity was
developed in order to obtaining compounds with a lower degree of inhibition of the key CYP 3A4 cytochrome.
Keywords: macrocycles, cytochromes CYP 3A4, tuberculocidal activity
DOI: 10.1134/S1070363217040107
Recently, we have performed a search for a universal
pharmacophore combination for serine/threonine kinase
class, and have shown a possibility of optimizing this
pharmacophore for individual types of kinases to
obtain active and selective compounds [1, 2]. As a
result of adapting the universal pharmacophore to the
mycobacterial serine/threonine kinase PknB, we have
obtained compounds 1–4 with pronounced bacterio-
static activity against the virulent strain Mycobacterium
tuberculosis (H37Rv) in a liquid medium [3] (Scheme 1).
Scheme 1.
N
N
N
N
N
S
N
S
N
N
HN
HN
S
N
N
1
2
H37Rv: MIC 2 μМ
H37Rv: MIC 3 μМ
CYP3A4: 95% at 10 μМ
CYP3A4: 95% at 10 μМ
S
N
N
N
N
HN
S
N
N
HN
N
N
NH
S
N
3
4
H37Rv: MIC 8 μМ
CYP3A4: 94% at 10 μМ
H37Rv: MIC 1 μМ
CYP3A4: 97% at 10 μМ
717

718
VASILEVICH et al.
Scheme 2.
N
N
5_6
5_6
N
NH
N
NH
N
N
O
N
S
N
S
S
O
R
R
O
O
N
H
N
H
A
B
Scheme 3.
O
O
O
O
O
O
(1) CDI, MeCN
(2) (NH4)2CO3
(1) Et3OBF4, CH2Cl2
(2) NH3, MeOH
O
O
NH
NH2
N
N
N
Me3N, 20oC
OH
NH2
n
n
n
5a, 5b
6a, 6b
7a, 7b
O
O
O
O
t-BuONa,
CH3C(O)CH2COOEt
N
N
N
POCl3, Me2NC6H5
N
N
toluene, 60oC
EtOH, Δ
N
OH
Cl
n
n
8a, 8b
9a, 9b
S
H2N
COOMe
O
O
O
O
O
N
N
N
N
O
HCl
H
N
N
S
S
dioxane
Хantphos,
Pd2(dba)3 CHCl3,
Na2CO3, MW, 140oC
N
N
N
N
N
H
n
H
n
10a, 10b
n = 1 (а), 2 (b).
11a, 11b
MIC is the minimum concentration of inhibition,
the concentration required to completely inhibit the
proliferation of H37Rv in a liquid medium.
etc. [4, 5]. In particular, this approach has been suc-
cessfully applied to improve the properties of kinase
inhibitors [6, 7]. The structural analysis of compounds
1–4 suggests that they will easily undergo the ring
closure by introducing a linker connecting the terminal
aromatic moieties. This work is aimed to design and
synthesize 15–16-membered macrocyclic compounds
that satisfy the requirements of the pharmacophore we
have found.
A disadvantage of these compounds was a high
lipophilicity and almost complete inhibition of the key
cytochrome CYP 3A4 at a concentration of 10 μM.
The macrocyclization of molecules allows improv-
ing their selectivity, solubility, permeability, metabolism,
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DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
719
Scheme 4.
DIAD, Ph3P, phthalimide,
O
OH
THF, 20oC
N
N
N
O
12b, 12c
13b, 13c
O
O
m-CPBA, CH2Cl2, 20oC
TFAA, CH2Cl2, 20oC
N
N
N
N
O
OH
O
O
15b, 15c
14b, 14c
Boc2O, CH2Cl2, 20oC
N2H4 H2O, EtOH, Δ
HO
HO
NH2
N Boc
N
N
H
16b, 16c
17b, 17c
DMP, CH2Cl2, 20oC
O
N Boc
N
H
18b, 18c
One of the suggested macrocyclic building blocks
A was designed by connecting the rings with a me-
thyleneaminocarbonyl linker –CH2NHC(O)–. Another
building block B contained all three necessary
aromatic rings; however, unlike the original non-cyclic
analogs and compounds including building block A
one of the aromatic rings was bound to the saturated
pyrrolidine or piperidine ring not by a methylene linker
but by two sulfamide linkers, which provided the
structure with an additional rigidity (Scheme 2).
solution in methanol. The condensation of amidine 7
with ethyl acetoacetate afforded bicyclic compound 8
which was further converted to chloro derivative 9 by
the action of phosphoryl chloride. This compound was
brought into a Pd-catalyzed Buchwald–Hartwig reac-
tion with 2-amino-5-carboxymethylthiazole to form com-
pound 10. Boc deprotection resulted in the formation
of compound 11.
Aldehyde 18a was obtained by Boc-protection of
commercial 3-(aminomethyl)benzaldehyde. Aldehydes
18b and 18c were prepared according to Scheme 4. In
the first stage the Mitsunobu reaction of the cor-
responding 6-methylpyridylmethanol 12 with phthal-
imide in the presence of diisopropyl azodicarboxylate
(DIAD) and triphenylphosphine afforded compound
13. Further oxidation of 13 with m-chloroperbenzoic
acid in methylene chloride gave rise to pyridinium
oxide 14 which was converted to alcohol 15 by the
treatment with trifluoroacetic anhydride (TFAA). The
removal of the phthalimide protecting group followed
by treatment with di-t-butyloxycarbonate resulted in
the formation of the corresponding Boc-derivative 17.
Dess–Martin oxidation of the latter afforded the
desired aldehyde 18.
A computer simulation confirmed the possibility of
superimposing the designed macrocyclic compounds
with the PknB-pharmacophore which we had found
earlier consisting of three hydrophobic regions and
projections of the hydrogen bond donor and acceptor
[1, 2].
Synthesis of compounds containing the building
block A was started from the preparation of inter-
mediate 11 (Scheme 3). To this end, Boc-proline 5a or
Boc-pipecolinic acid 5b was converted to the
corresponding amide 6 and then to amidine 7 by
successive treatment with an equimolar amount of
triethyloxonium tetrafluoroborate and a 10% ammonia
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720
VASILEVICH et al.
Scheme 5.
O
(1) IBCF, THF
PCC, CH2Cl2, 20oC
(2) NaBH4, EtOH
OH
OH
O
Hlg
Hlg
Hlg
HN
HN
HN
O
O
O
O
O
O
18d, 18e
19d, 19e
20d, 20e
O
O
CH3I, NaH, THF
OH
OH
F
F
HN
N
O
O
O
O
19d
19f
(1) IBCF, THF
(2) NaBH4, EtOH
PCC, CH2Cl2, 20oC
OH
O
F
F
N
N
O
O
20f
O
O
18f
Hlg = F (d), Cl (e).
Aldehydes 18d–18f were prepared by reduction of
the corresponding benzoic acids 19 to alcohols 20
followed by selective oxidation with pyridinium
chlorochromate (Scheme 5). To prepare aldehyde 18f
acid 19b was initially alkylated with methyl iodide
before reduction.
Compounds containing the building block B were
prepared as shown in Scheme 7. Initially, the inter-
mediate 11a reacted with the corresponding sulfonyl
chloride 27a–27c in the presence of triethylamine to
give compounds 28a–28c. Further removal of the
protective triflate group followed by intramolecular
acylation in the presence of TBTU led to the formation
of macrocyclic compounds 30a–30c.
The synthesis of the macrocyclic compounds con-
taining the building block A is given in Scheme 6. The
reductive amination of intermediate 11 and the
corresponding aldehyde 18 led to the formation of the
protected amino acid 21. Further deprotection and
cyclization in the presence of O- (benzotriazol-1-yl)-
N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU)
resulted in the formation of target macrocycles 23 and
26 with yields of 35–88%.
The resulting macrocyclic compounds were tested
for inhibitory activity against liver cytochromes
CYP450. The data obtained showed that in the vast
majority of cases macrocyclization resulted in a signi-
ficant decrease in inhibition of the key cytochrome
CYP3A4 and cytochrome CYP2D6 (see table).
However, an unambiguous conclusion about the effect
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 87 No. 4 2017

DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
721
Scheme 6.
Boc
HN
Hlg
Х
N
H
N
Hlg
NaBH(OAc)3, CH2Cl2, 20oC
Х
N
N
NH
S
+
N
n
HN
N
N
NH
S
Boc
O
n
N
O
O
O
O
18a_18f
NH2
21a_21f, 24a_24f
11a, 11b
N
Hlg
Х
n
N
NH
HCl, NaOH,
H2O_MeOH, 20oC
TBTU, Et3N, 20oC
N
N
S
N
N
N
NH
S
Х
n
O
N
NH
Hlg
O
23a_23f, 26a_26f
HO
22a_22f, 25a_25f
n = 1 (11а, 21а–23f), 2 (11b, 24а–26f); Х = СН (а, d–f), N (b, c); Hlg = 0 (а–c), F (d, f), Cl (e).
of the structure on the inhibition of cytochromes is
difficult to make. In some cases, the pyrrolidine ring
(23d) was preferable to the piperidine (26d). At the
same time, the inhibitory effect of compounds with the
piperidine ring 26a–26c exceeds those of analogues
23a–23c containing the pyrrolidine ring. The methyl-
tion of the amide linker (26d, 26f, 30a) resulted in a
significant decrease in the inhibition of all cyto-
chromes. In general, compounds containing the
building block A seem preferable to compounds with
the building block B, but these differences are not
significant. The best compounds in the inhibition
parameter of liver cytochromes were compounds 26a
and 26f.
instrument using chemical ionization at atmospheric
pressure (ZORBAX Eclipse XDB-C18 column, 2.1×
15 mm, 1.8 μm, flow rate 750 μL/min, column tem-
perature 25°C; mobile phase: A, 0.1% aqueous
solution of formic acid; B, acetonitrile; detection with
a photodiode array (PDA), 200–800 nm, photodiode
array detector over the entire ultraviolet and visible
range from 200 to 800 nm). H NMR spectra were
recorded on a MERCURY plus 400 MHz Varian
spectrometer, internal reference tetramethylsilane.
1
tert-Butyl 2-carbamoylpyrrolidine-1-carboxylate
(6a). To a solution of 20 g (93 mmol) of Boc-proline
5a in 150 mL of anhydrous acetonitrile were added
successively 19.5 g (120 mmol) of 1,1'-carbonyl-
diimidazole, 21.0 mL (150 mmol) of trimethylamine,
and 19.2 g (200 mmol) of ammonium carbonate. The
reaction mixture was stirred at room temperature for
24 h. After the reaction completion the mixture was
EXPERIMENTAL
Chromato-mass spectrometric analysis was performed
on a Surveyor MSQ (Thermo Fisher Scientific)
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 87 No. 4 2017

722
VASILEVICH et al.
Scheme 7.
O
N
F
F
O
O
F
Cl
n
R
O
O
S
O
N
O
H
Me3N, MeCN, 20°C
O
N
+
S
O
S
N
O
N
n
O
F
N
H
N
S
N
N
O
R
N
F
N
N
F
H
28a_28c
27a_27c
11a
O
O
O
NH
R
O
O
n
N
n
S
R
N
O
NaOH, MeOH
HN
TBTU, Me3N, ТHF
O
S
O
N
OH
S
N
N
S
N
N
N
H
N
N
30a_30c
29a_29c
n = 1, R = Me (а); n = 2, R = Н (b); n = 3, R = Н (c).
poured into a saturated solution of potassium carbonate
(250 mL) and extracted with dichloromethane (3 ×
250 mL). The combined organic extracts were washed
with water (2 × 150 mL) and dried with sodium
sulfate. After removal of the solvent the reaction
product was purified by flash chromatography on silica
gel using hexane–ethyl acetate (100 : 0 → 1 : 1). Yield
12.3 g (62%). Mass spectrum, m/z (Irel, %): 215.1 (100)
[M + H]+.
tert-Butyl 2-(4-hydroxy-6-methylpyrimidin-2-yl)
pyrrolidine-1-carboxylate (8a). A solution of 5.9 g
(61 mmol) of sodium t-butoxide in 150 mL of
anhydrous ethanol was added to a solution of 16.3 g
(54.5 mmol) of amidine 7a in ethanol. The resulting
mixture was stirred for 10 min, and then 11.7 g
(90 mmol) of ethyl acetoacetic acid was added. The
reaction mixture was boiled for 10 h, then cooled to
room temperature, poured into a saturated solution of
ammonium chloride (300 mL), and extracted with
dichloromethane (2 × 200 mL). The combined extracts
were dried with sodium sulfate, and the solvent was
tert-Butyl 2-carbamimidoylpyrrolidine-1-carboxy-
late (7a). Triethyloxonium tetrafluoroborate (14.6 g,
77 mmol) was added with stirring to a solution of
12.3 g (57.4 mmol) of amide 6a in 200 mL of
anhydrous dichloromethane. The reaction mixture was
stirred for 3 h (control by TLC, ethyl acetate–dichloro-
methane, 1 : 2). The solvent was distilled off in a
vacuum at ≤40 °C. The oily residue was treated with
10% ammonia in methanol (150 mL) for 24 h at room
temperature. After the removal of methanol the residue
was triturated with diethyl ether, then ether was
decanted, and the residue was dried at room
temperature. Yield 16.3 g (95%). Mass spectrum, m/z
(Irel, %): 214.1 (100) [M + H]+. Amidine 7a was used
in the next step without further purification.
distilled off in
a
vacuum. The residue was
chromatographed on a silica gel column eluting with a
1 : 1 mixture of ethyl acetate and dichloromethane.
1
Yield 8.7 g (57%). H NMR spectrum (400 MHz,
DMSO-d6), δ, ppm: 1.15 s [9H, (CH3)C], 1.73–1.95 m
(2H, CH2CH2), 2.06 s (3H, CH3C=N), 2.21 m (2H,
CH2CH2CH), 3.34 m (1H, CH2CH2N), 3.47 m (1H,
CH2CH2N), 4.40 m (1H, CH2CHN), 5.99 s (1H, CH=C),
12.15 br.s (1H, OH). Mass spectrum, m/z (Irel, %):
280.2 (100) [M + H]+. Found, %: C 60.28; H 7.45; N
15.02. C14H21N3O2. Calculated, %: C 60.20; H 7.58; N
15.04.
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DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
723
Inhibitory effectа of compounds 23a–23f, 26a–26f, and 30a–30c on human liver cytochromes CYP450
CYP3A4, %
(IC50, μM)
CYP1A2, %
(IC50, μM)
CYP2C9, %
(IC50, μM)
CYP2D6, %
(IC50, μM)
CYP2C19, %
(IC50, μM)
Compound
1
95 (0.536)
38
65
83 (1.543)
71 (3.708)
2
95 (0.634)
21
38
69
35
3
94 (0.810)
38
51
92 (1.49)
54
4
96 (0.723)
33
30
82 (2.132)
29
23а
23b
23c
23d
23e
23f
26а
26b
26c
26d
26e
26f
30а
30b
87
–7
14
4
14
97 (0.366)
40
26
46
9
73
16
12
–1
–5
62
13
50
24
70 (4.179)
76
28
19
5
76 (3.161)
88
53
24
56
26
58
3
4
3
5
75
2
17
–4
84 (1.636)
62
7
1
80 (1.226)
14
99 (0.381)
92 (0.578)
65
71 (1.562)
80% (3.691)
39
49
68
11
9
69
18
14
1
3
86
15
3
83 (1.612)
18
10
49
59
–4
97 (0.158)
78
30c
98 (0.508)
78
89 (1.884)
87 (0.157)
a At с = 10 μM.
tert-Butyl 2-(4-chloro-6-methylpyrimidin-2-yl)pyr-
rolidine-1-carboxylate (9a). N,N-Dimethylaniline (37.0 g,
306 mmol) and phosphoryl chloride (15.6 g,
102 mmol) were successively added to a solution of
8.7 g (31 mmol) of compound 8a in 200 mL of
anhydrous toluene. The mixture was refluxed for 3 h,
then cooled to room temperature, and left overnight.
The reaction mixture was poured into cold water
(300 mL), the organic layer was separated, washed
with 1 N HCl (2 × 70 mL) and water (1 × 70 mL),
dried with sodium sulfate, and evaporated. The reac-
tion product was isolated on a silica gel column eluting
with a 1 : 3 mixture of ethyl acetate and hexane. Yield
6.2 g (67%). Mass spectrum, m/z (Irel, %): 297.9 (100)
[M + H]+.
9a, 0.95 g (6 mmol) of 2-amino-5-carbomethoxy-
thiazole, 4 mol% of 9.9-dimethyl-4,5-bis(diphenyl-
phosphino)xanthene, 2 mol% of Pd2(dba)3·CHCl3 and
0.95 g (9 mmol) of sodium carbonate in 15 mL of a
toluene–water (10 : 1) mixture was stirred under argon
in a microwave oven at 140 °C for 2 h. After cooling
to room temperature 30 mL water was added to the
reaction mixture, then the product was extracted with
ethyl acetate (2 × 30 ml). The combined organic
extracts were dried with sodium sulfate, and the
solvent was distilled off in a vacuum. The residue was
purified by column chromatography on silica gel (ethyl
1
acetate–hexane, 1 : 2). Yield 2.02 g (80%). H NMR
spectrum (400 MHz, DMSO-d6), δ, ppm: 1.39–0.85 m
[9H, (CH3)C], 1.97–1.73 m (3H, CH2CH2), 2.32 m
(1H, CH2CH2CH), 2.36 s (3H, CH3C=N), 3.47 m (1H,
CH2CH2N), 3.79–3.68 m (1H, CH2CH2N), 3.81 s (3H,
Methyl 2-[2-(1-t-butyloxycarbonylpyrrolidin-2-
yl)-6-methylpyrimidin-4-ylamino]thiazole-5-carboxy-
late (10a). A mixture of 2.0 g (6.7 mmol) of chloride
3
CH3O), 4.72 d (1H, CH2CHN, JHH = 13.7 Hz), 6.76 d
3
(1H, CCH=C, JHH = 12.1 Hz), 8.07 s (1H, C=CHN),
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 87 No. 4 2017

724
VASILEVICH et al.
11.99 s (1H, NH). Mass spectrum, m/z (Irel, %): 420.3
(100) [M + H]+.
Methyl 2-(6-methyl-2-piperidin-2-ylpyrimidin-4-
ylamino)thiazole-5-carboxylate (11b). The yield was
quantitative. Mass spectrum, m/z (Irel, %): 334.2 (100)
[M + H]+.
Methyl 2-(6-methyl-2-pyrrolidin-2-yl-pyrimidin-
4-ylamino)thiazole-5-carboxylate (11a). To a solu-
tion of 2.02 g (4.8 mmol) of the Boc derivative 10a in
20 mL of methanol was added 16% (by weight) of a
solution of hydrogen chloride in 20 mL of dioxane.
The mixture was stirred for 5 h at room temperature.
Then the solvent was distilled off in a vacuum, the
residue was alkalinized with 10% sodium hydroxide
solution. The precipitate was filtered off, washed with
water, and dried. The yield was quantitative. Mass
spectrum, m/z (Irel, %): 320.4 (100) [M + H]+.
2-[(6-Methyl-2-pyridyl)methyl]isoindoline-1,3-
dione (13b). To a solution of 4.1 g (33.7 mmol) of (6-
methyl-2-pyridyl)methanol 12b in 200 mL of THF was
added 9.69 g (37 mmol) of triphenylphosphine and
4.95 g (33.7 mmol) of phthalimide. The mixture was
cooled to 0 °C and a solution of 7.47 g (37 mmol) of
DIAD in 20 mL of THF was added. The mixture was
stirred at room temperature for 8 h. The solvent was
distilled off in a vacuum, and the residue was
chromatographed on a silica gel column (ethyl acetate–
hexane, 1 : 2). Yield 8 g (96%). Mass spectrum, m/z
(Irel, %): 253.2 (100) [M + H]+.
Compound 11b was prepared similarly to com-
pound 11a.
tert-Butyl 2-carbamoylpiperidine-1-carboxylate
(6b) was prepared from 23.0 g (100 mmol) of Boc-
pipecoline-2-carboxylic acid 5b. Yield 17.6 g (77%).
Mass spectrum, m/z (Irel, %): 229.1 (100) [M + H]+.
6-Methyl-2-(1,3-dioxoisoindolin-2-yl)methylpyri-
dine-1-oxide (14b). m-Chloroperbenzoic acid (7 g,
39 mmol) was added to a solution of 6.6 g (26 mmol)
of compound 13b in 200 mL of anhydrous dichloro-
methane with cooling to 0 °C. The mixture was stirred
at room temperature for 2 h and then poured into a
saturated potassium carbonate solution (200 mL). The
organic layer was separated, washed with water (1 ×
100 mL), dried with sodium sulfate, and evaporated.
The residue was recrystallized from hexane. Yield
5.86 g (84%). Mass spectrum, m/z (Irel, %): 269.2 (100)
[M + H]+.
tert-Butyl 2-carbamimidoylpiperidine-1-carboxy-
late (7b) was prepared from 17.6 g (77 mmol) of
amide 6b. Yield 19.2 g (79%). Mass spectrum, m/z
(Irel, %): 228.1 (100) [M + H]+.
tert-Butyl 2-(4-hydroxy-6-methylpyrimidin-2-yl)-
piperidine-1-carboxylate (8b). Yield 11.4 g (64%).
Mass spectrum, m/z (Irel, %): 294.2 (100) [M + H]+.
tert-Butyl 2-(4-chloro-6-methylpyrimidin-2-yl)-
piperidine-1-carboxylate (9b). Yield 70%. H NMR
2-[(6-Hydroxymethyl-2-pyridyl)methyl]isoindoline-
1,3-dione (15b). To a solution of 5.86 g (21.8 mmol)
of compound 14b in 100 mL of anhydrous THF was
added 7.64 mL (55 mmol) of trifluoroacetic anhydride.
The reaction mixture was stirred for 3 h. After the
reaction completion the solvent was distilled off in a
vacuum, the residue was dissolved in dichloromethane
(100 mL) and saturated potassium carbonate solution
(100 mL) was added. The reaction mixture was
vigorously stirred for 3 h, then the organic layer was
separated, dried with sodium sulfate, and evaporated.
The residue was chromatographed on a silica gel
column (ethyl acetate–hexane, 1 : 1). Yield 1.75 g
(30%). Mass spectrum, m/z (Irel, %): 269.2 (100)
[M + H]+.
1
spectrum (400 MHz, CDCl3), δ, ppm: 1.11 m (1H,
NCH2CH2CH2), 1.36 s [9H, (CH3)C], 1.39–1.67 m
(3H, NCH2CH2CH2), 1.86 m (1H, CHCH2), 2.41 s
(3H, CH3C=N), 2.45–2.67 m (3H, NCH2 + CHCH2),
5.29 m (1H, NCH), 7.02 s (1H, CH=CCl). Mass
spectrum, m/z (Irel, %): 311.9 (100) [M + H]+.
Methyl 2-[2-(1-t-butyloxycarbonylpiperidin-2-yl)-
6-methylpyrimidin-4-ylamino]thiazole-5-carboxylate
1
(10b). Yield 82%. H NMR spectrum (400 MHz,
DMSO-d6), δ, ppm: 1.02–1.22 m (2H, NCH2CH2CH2),
1.48 s [9H, (CH3)C], 1.56 t (2H, NCH2CH2CH2, J =
13.7 Hz), 1.87–1.77 m (2H, CH2), 2.35 s (3H,
3
CH3C=N), 2.58 d (1H, NCH2CH2, JHH = 13.0 Hz),
3.79 s (3H, CH3O), 3.95 t (1H, NCH2CH2, J = 12.1 Hz),
5.18 m (1H, NCH), 6.78 s (1H, CH, pyrimidine), 8.09
s (1H, thiazole), 12.00 s (1H, NH). Mass spectrum, m/z
(Irel, %): 434.2 (100) [M + H]+. Found, %: C 55.48; H
6.40; N 16.07. C20H27N5O4S. Calculated, %: C 55.41;
H 6.28; N 16.15.
[6-(Aminomethyl)-2-pyridyl]methanol (16b). To
a solution of 1.75 g (6.5 mmol) of compound 15b in
100 mL of ethanol was added 0.325 g (6.5 mmol) of
hydrazine hydrate. The mixture was boiled for 4 h and
then cooled to room temperature. The precipitate was
filtered off, the filtrate was evaporated to dryness to
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DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
725
yield 0.9 g (100%) of compound 16b, which was used
in the next step without further purification. Mass
spectrum, m/z (Irel, %): 139.1 (100) [M + H]+.
Ar-CH2NH), 5.48 br.s (1H, NH), 7.42–7.48 m (1H,
CH2, pyrimidine), 7.94–8.05 m (1H, CH2, pyrimidine),
9.43 s (1H, CH2, pyrimidine), 10.16 s (1H, CHO).
Mass spectrum, m/z (Irel, %): 236.98 (100) [M + H]+.
tert-Butyl N-[(6-hydroxymethyl-2-pyridyl)methyl]-
carbamate (17b). To a solution of 0.9 g (6.5 mmol) of
compound 16b in 50 mL of dichloromethane was
added 1.53 g (7 mmol) of Boc2O. The mixture was
stirred at room temperature for 4 h. The solvent was
distilled off in a vacuum and the residue was purified
by flash chromatography on silica gel (ethyl acetate–
hexane, 1 : 1). Yield 1.43 g (93%). Mass spectrum, m/z
(Irel, %): 239.0 (100) [M + H]+.
tert-Butyl (2-fluoro-5-hydroxymethylbenzyl)carb-
aminate (20d). To a mixture of 538 mg (2.0 mmol) of
3-(tert-butoxycarbonylaminomethyl)-4-fluorobenzoic
acid 19d and 222 mg (2.2 mmol) of triethylamine in
30 mL of anhydrous THF at 0 °C was added dropwise
300 mg (2.2 mmol) of isobutyl chloroformate. The
reaction mixture was stirred at room temperature for
1.5 h. Triethylamine hydrochloride was filtered off and
washed with a small amount of cold THF. To the
resulting solution of the mixed anhydride was added
380 mg (10 mmol) of sodium borohydride and 1 mL of
ethanol. The suspension was stirred at room tem-
perature for 1 h, then poured into water (100 mL) and
extracted with dichloromethane (2 × 100 mL). The
organic phase was evaporated, the residue was purified
on a silica gel column eluting with dichloromethane–
ethyl acetate. Yield 393 mg (77%). Mass spectrum,
m/z (Irel, %): 256.20 (100) [M + H]+.
tert-Butyl N-[(6-formyl-2-pyridyl)methyl]carba-
mate (18b). To a solution of 1.43 g (6 mmol) of
alcohol 16b in 100 mL of dichloromethane was added
3.05 g (7.2 mmol) of Dess–Martin periodinane. The
resulting suspension was stirred for 48 h. The
precipitate was filtered off, the solvent was distilled off
in a vacuum and the residue was purified by
chromatography on silica gel (ethyl acetate–hexane,
1
1 : 4). Yield 1.20 g (85%). H NMR spectrum
(400 MHz, CDCl3), δ, ppm: 1.47 s [9H, (CH3)3CO],
3
4.53 d (2H, Ar-CH2NH, JHH = 5.6 Hz), 5.49 br.s (1H,
tert-Butyl (2-fluoro-5-formylbenzyl)carbaminate
(18d). Pyridinium chlorochromate (473 mg, 2.2 mmol)
was added to a solution of 380 mg (1.5 mmol) of
compound 20d in 20 mL of anhydrous dichloro-
methane. The resulting mixture was stirred for 1.5 h at
room temperature. The solution was decanted, the
solvent was distilled off in a vacuum, and the residue
was purified by flash chromatography on silica gel
NH), 7.49–7.53 m (1H, CH2, pyrimidine), 7.82–7.87 m
(2H, CH2, pyrimidine), 10.07 s (1H, CHO). Mass spec-
trum, m/z (Irel, %): 236.95 (100) [M + H]+.
Aldehyde 18c was prepared similarly to compound
18b.
2-[(6-Methyl-3-pyridyl)methyl]isoindoline-1,3-
dione (13c). Yield 65%. Mass Spectrum, m/z
(Irel, %): 253.2 (100) [M + H]+.
1
(dichloromethane). Yield 340 mg (90%). H NMR
spectrum (400 MHz, CDCl3), δ, ppm: 1.37 s [9H,
(CH3)3CO], 4.22 d (2H, Ar-CH2NH), 5.48 br.s (1H,
NH), 7.32–7.42 m (2H, CH2, pyrimidine), 7.83–7.90 m
(1H, CH2, pyrimidine), 9.95 s (1H, CHO). Mass
spectrum, m/z (Irel, %): 253.98 (100) [M + H]+.
6-Methyl-3-(1,3-dioxoisoindolin-2-yl)methylpyri-
dine-1-oxide (14c). Yield 45%. Mass spectrum, m/z
(Irel, %): 269.12 (100) [M + H]+.
2-[(6-Hydroxymethyl-3-pyridyl)methyl]isoindo-
line-1,3-dione (15c). Yield 35%. Mass spectrum, , m/z
(Irel, %): 269.2 (100) [M + H]+.
Aldehyde 18e was prepared similarly to compound
18d.
tert-Butyl (2-chloro-5-hydroxymethylbenzyl)carb-
aminate (20e). Yield 87%. Mass spectrum, m/z (Irel,
%): 272.90 (100) [M + H]+.
[6-(Aminomethyl)-3-pyridyl]methanol (16c). The
yield was quantitative. Mass spectrum, m/z (Irel, %):
139.2 (100) [M + H]+.
tert-Butyl (2-chloro-5-formylbenzyl)carbaminate
(18e). Yield 93%. H NMR spectrum (400 MHz,
tert-Butyl N-[(6-hydroxymethyl-3-pyridyl)methyl]-
carbamate (17c). Yield 95%. Mass spectrum, m/z (Irel,
%): 239.0 (100) [M + H]+.
1
DMSO-d6), δ, ppm: 1.44 s [9H, (CH3)3CO)], 4.25 d
3
(2H, Ar-CH2NH, JHH = 6.1 Hz), 5.41 br.s (1H, NH),
3
tert-Butyl N-[(6-formyl-3-pyridyl)methyl]carba-
mate (18c). Yield 91%. 1H NMR spectrum (400 MHz,
CDCl3), δ, ppm: 1.48 s [9H, (CH3)3CO)], 4.53 m (2H,
7.66 d (1H, CH2, pyrimidine, JHH = 8.0 Hz), 7.79 m
(2H, CH2, pyrimidine), 9.99 s (1H, CHO). Mass spec-
trum, m/z (Irel, %): 270.77 (100) [M + H]+.
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726
VASILEVICH et al.
3-[(t-Butyloxycarbonylmethylamino)methyl]-4-
solution. The resulting mixture was stirred at room
temperature for 6 h, then neutralized with concentrated
hydrochloric acid to pH 6, and evaporated to dryness.
The resulting amino acid was used in the next step
without further purification. The yield was quanti-
tative. Mass spectrum, m/z (Irel, %): 519.6 (100) [M + H]+.
fluorobenzoic acid (19f). Sodium hydride (480 mg,
12.0 mmol, 60% in mineral oil) was added to a
solution of 1.24 g (4.6 mmol) of acid 19d in 100 mL of
anhydrous THF. The resulting mixture was stirred for
15 min and then 1.70 g (12.0 mmol) of methyl iodide
was added. The reaction mixture was stirred for 16 h at
room temperature, after which 10 mL of a 5% aqueous
solution of NaOH was added and stirring was
continued for an additional 12 h. After the completion
of the reaction the mixture was acidified to pH 5 with a
solution of citric acid, extracted with dichloromethane
(2 × 150 mL), and evaporated. The residue was
triturated with hexane, then filtered off, and dried.
Yield 1.19 g (92%). Mass spectrum, m/z (Irel, %):
283.99 (100) [M + H]+.
2-(2-{1-[3-(2-Aminoethyl)-4-methoxybenzene-
sulfonyl]pyrrolidin-2-yl}-6-methylpyrimidin-4-yl-
amino)thiazole-5-carboxylic acid (29b) was obtained
similarly with quantitative yield. Mass spectrum, m/z
(Irel, %): 519.4 (100) [M + H]+.
2-(2-{1-[3-(2-Aminopropyl)-4-methoxybenzene-
sulfonyl]pyrrolidin-2-yl}-6-methylpyrimidin-4-yl-
amino)thiazole-5-carboxylic acid (29c) was obtained
similarly with quantitative yield. Mass spectrum, m/z
(Irel, %): 532.6 (100) [M + H]+.
tert-Butyl (2-fluoro-5-hydroxymethylbenzyl)methyl-
carbaminate (20f) was obtained analogously to
compound 20d. Yield 69%. Mass spectrum, m/z (Irel,
%): 270.30 (100) [M + H]+.
General procedure for the synthesis of macro-
cycles 23a–23f and 26a–26f.
Synthesis of compounds 21a–21f and 24a–24f. To
a suspension of 1.00 mmol of compound 11a or 11b in
20 mL of dichloromethane was added 1.20 mmol of
the corresponding aldehyde 18a–18f. The mixture was
stirred for 10–15 min, and then 252 mg (1.20 mmol) of
sodium triacetoxyborohydride was added. The reaction
mixture was stirred at room temperature for 12 h, then
poured into 30 mL of a saturated solution of potassium
carbonate and extracted with dichloromethane
(50 mL). The organic layer was separated, dried with
sodium sulfate, and evaporated. The residue was
purified on a silica gel column (ethyl acetate–
dichloromethane, 1 : 2).
tert-Butyl (2-chloro-5-formylbenzyl)methyl carba-
mate (18f) was obtained analogously to compound
18d. Yield 97%. 1H NMR spectrum (400 MHz, DMSO-
d6), δ, ppm: 1.30 s [9H, (CH3)3CO)], 2.82 s (3H, CH3N),
4.46 s (2H, Ar-CH2NH), 7.42 d.d (1H, CH2,
3
pyrimidine, JHH = 9.6, 8.7 Hz), 7.79 d (1H, CH2,
3
pyrimidine, JHH = 6.8 Hz), 7.88–7.96 m (1H, CH2,
pyrimidine), 9.91 s (1H, CHO). Mass spectrum, m/z
(Irel, %): 268.27 (100) [M + H]+.
2-{2-[1-(4-Methoxy-3-{[methyl-(2,2,2-trifluoro-
acetyl)amino]methyl}benzenesulfonyl)pyrrolidin-2-
yl]-6-methylpyrimidin-4-ylamino}thiazole-5-carbo-
xylic acid (28a). 4-Methoxy-3-{[methyl-(2,2,2-tri-
fluoroacetyl)amino]methyl}benzenesulfonyl chloride
27a (345 mg, 1 mmol) and triethylamine (0.17 mL,
1.2 mmol) were added to a solution of 319 mg
(1 mmol) of compound 11a in 10 mL of anhydrous
acetonitrile. The mixture was stirred for 4 h, then
poured into a saturated solution of potassium carbonate
(20 mL), and the product was extracted with dichloro-
methane. The extract was dried with sodium sulfate.
After removing the solvent, the residue was triturated
with hexane, filtered off, and dried. Yield 500 mg
(80%). Mass spectrum, m/z (Irel, %): 629.6 (100) [M + H]+.
Methyl 2-(2-{1-(3-[(tert-butoxycarbonylamino)-
methyl]benzyl}pyrrolidin-2-yl)-6-methylpyrimidin-
4-ylamino)thiazole-5-carboxylate (21a). Yield 51%.
Mass spectrum, m/z (Irel, %): 538.2 (100) [M + H]+.
Methyl 2-{2-[1-({6-[(tert-butoxycarbonylamino)
methyl]pyridin-2-yl}methyl)pyrrolidin-2-yl]-6-
methylpyrimidin-4-ylamino}thiazole-5-carboxylate
(21b). Yield 74%. Mass spectrum, m/z (Irel, %): 539.3
(100) [M + H]+.
Methyl 2-{2-[1-({5-[(tert-butoxycarbonylamino)-
methyl]pyridin-2-yl}methyl)pyrrolidin-2-yl]-6-
methylpyrimidin-4-ylamino}thiazole-5-carboxylate
(21c). Yield 78%. Mass spectrum, m/z (Irel, %): 539.2
(100) [M + H]+.
2-{2-[1-(4-Methoxy-3-methylaminomethylbenzene-
sulfonyl)pyrrolidin-2-yl]-6-methylpyrimidin-4-yl-
amino}thiazole-5-carboxylic acid (29a). To a solu-
tion of 500 mg (0.8 mmol) of compound 28a in 10 mL
of methanol was added 5 mL of saturated NaOH
Methyl 2-[2-(1-{3-[(tert-butoxycarbonylamino)-
methyl]-4-fluorobenzyl}pyrrolidin-2-yl)-6-methyl-
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DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
727
pyrimidin-4-ylamino]thiazole-5-carboxylate (21d).
Yield 84%. Mass spectrum, m/z (Irel, %): 556.3 (100)
[M + H]+.
and a solution of 200 mg (5.00 mmol) of NaOH in
3 mL of water was added. The resulting mixture was
stirred at room temperature for 18 h and then acidified
with hydrochloric acid to pH 5–6. After the solvent
removal the residue was dried in air at 60°C for 6 h.
Amino acid 22 or 25 with an admixture of inorganic
salts was prepared, which was used in the next step
without further purification.
Methyl 2-[2-(1-{3-[(tert-butoxycarbonylamino)-
methyl]-4-chlorobenzyl}pyrrolidin-2-yl)-6-methyl-
pyrimidin-4-ylamino]thiazole-5-carboxylate (21e).
Yield 83%. Mass spectrum, m/z (Irel, %): 572.3 (100)
[M + H]+.
Synthesis of macrocycles 23a-–23f and 26a–26f.
To a suspension of 0.5 mmol of compound 22 or 25 in
150 mL of dichloromethane was added 280 μL
(2.00 mmol) of triethylamine and 193 mg (0.60 mmol)
of TBTU. The mixture was stirred at room temperature
for 24 h, then washed with a solution of potassium
carbonate, and dried with sodium sulfate. After
removal of the solvent the residue was chromato-
graphed on a silica gel column (dichloromethane–
methanol, 30 : 1).
Methyl 2-{2-[1-(3-{[tert-butoxycarbonyl(methyl)-
amino]methyl}-4-fluorobenzyl)pyrrolidin-2-yl]-6-
methylpyrimidin-4-ylamino}thiazole-5-carboxylate
(21f). Yield 88%. Mass spectrum, m/z (Irel, %): 570.3
(100) [M + H]+.
Methyl 2-[2-(1-{3-[(tert-butoxycarbonylamino)-
methyl]benzyl}piperidin-2-yl)-6-methylpyrimidin-4-
ylamino]thiazole-5-carboxylate (24a). Yield 66%.
Mass spectrum, m/z (Irel, %): 552.4 (100) [M + H]+.
1
Methyl 2-{2-[1-({6-[(tert-butoxycarbonylamino)-
methyl]pyridin-2-yl}methyl)piperidin-2-yl]-6-methyl-
pyrimidin-4-ylamino}thiazole-5-carboxylate (24b).
Yield 66%. Mass spectrum, m/z (Irel, %): 553.5 (100)
[M + H]+.
Macrocycle 23a. Yield 41%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.77–1.89 m (2H, CH2,
pyrrolidine), 1.94–2.01 m (1H, CH2, pyrrolidine), 2.18
d.d (1H, CH2, pyrrolidine, 3JHH = 18.5, 9.8 Hz), 2.33 d
3
(3H, CH3-pyrimidine, JHH = 13.0 Hz), 3.08 d.d (2H,
CH2, pyrrolidine, 3JHH = 16.3, 8.0 Hz), 3.50 d (1H, CH,
Methyl 2-{2-[1-({5-[(tert-butoxycarbonylamino)-
methyl]pyridin-2-yl}methyl)piperidin-2-yl]-6-methyl-
pyrimidin-4-ylamino}thiazole-5-carboxylate (24c).
Yield 70%. Mass spectrum, m/z (Irel, %): 553.5 (100)
[M + H]+.
3
3
pyrrolidine, JHH = 13.3 Hz), 3.86 d (1H, CH2, JHH
=
3
13.3 Hz), 3.95 d.d (1H, CH2, JHH = 7.8, 4.4 Hz), 4.39
d.d.d (2H, CH2, 3JHH = 35.5, 16.3, 6.9 Hz), 6.68 s (1H,
3
pyrimidine), 7.08 d (1H, CH, Ph, JHH = 6.8 Hz), 7.21
d.d (2H, CH, Ph, 3JHH = 10.7, 7.3 Hz), 7.75 s (1H, CH,
Ph), 7.98 s [1H, C(O)NH], 8.46 t (1H, CH, thiazole,
3JHH = 6.6 Hz). Mass spectrum, m/z (Irel, %): 407.2
(100) [M + H]+. Found, %: C 62.15; H 5.49; N 20.48.
C21H22N6O. Calculated, %: C 62.05; H 5.45; N 20.67.
Methyl 2-[2-(1-{3-[(tert-butoxycarbonylamino)-
methyl]-4-fluorobenzyl}piperidin-2-yl)-6-methyl-
pyrimidin-4-ylamino]thiazole-5-carboxylate (24d).
Yield 54%. Mass spectrum, m/z (Irel, %): 570.4 (100)
[M + H]+.
1
Macrocycle 23b. Yield 35%. H NMR spectrum
Methyl 2-[2-(1-{3-[(t-butoxycarbonylamino)methyl]-
4-chlorobenzyl}piperidin-2-yl)-6-methylpyrimidin-
4-ylamino]thiazole-5-carboxylate (24e). Yield 64%.
Mass spectrum, m/z (Irel, %): 586.3 (100) [M + H]+.
(400 MHz, DMSO-d6), δ, ppm: 1.64–1.92 m (3H, CH2,
pyrrolidine), 2.29 d.d (1H, CH2, pyrrolidine, JHH
19.4, 9.0 Hz), 2.36 s (3H, CH3-pyrimidine), 2.87 m
(1H, CH2, pyrrolidine), 3.58–3.33 m (2H, CH2), 3.97 d
(1H, CH2, pyrrolidine, JHH = 21.0 Hz), 4.37 d.d (2H,
3
=
3
Methyl 2-{2-[1-(3-{[tert-butoxycarbonyl(methyl)-
amino]methyl}-4-fluorobenzyl)piperidin-2-yl]-6-
methylpyrimidin-4-ylamino}thiazole-5-carboxylate
(24f). Yield of 57%. Mass spectrum, m/z (Irel, %): 584.5
(100) [M + H]+.
3
CH2NH + CH, pyrrolidine, JHH = 17.1, 3.1 Hz), 4.76
d.d (1H, CH2NH, 3JHH = 17.2, 3.1 Hz), 6.70 s (1H, CH,
pyrimidine), 7.24 d.d (2H, CH, Ph, 3JHH = 11.0, 7.7 Hz),
3
7.70 t (1H, CH, Ph, JHH = 7.7 Hz), 7.80 s (1H, NH),
8.10 s (1H, thiazole), 11.69 s (1H, NH). Mass spec-
trum, m/z (Irel, %): 408.3 (100) [M + H]+. Found, %: C
59.02; H 5.29; N 23.91. C20H21N7OS. Calculated, %: C
58.95; H 5.19; N 24.06.
Synthesis of compounds 22a–22f and 25a–25f. To
a solution of 0.5 mmol of compound 21 or 24 in 5 mL
of ethanol was added 2 mL of concentrated hydro-
chloric acid. The mixture was stirred at room tem-
perature for 12 h. The solvents were evaporated to
dryness, the residue was dissolved in 3 mL of ethanol
1
Macrocycle 23c. Yield 35%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.85–2.26 m (4H, CH2,
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VASILEVICH et al.
pyrrolidine), 2.24–2.40 m (3H, CH3-pyrimidine), 2.91–
3.23 m (2H, CH2, pyrrolidine), 3.61–3.86 m (1H, CH2,
pyrrolidine), 3.84–4.53 m (4H, CH2 + CH2NH), 6.55 s
(1H, CH, pyrimidine), 7.29 m (1H, CH, Ph), 7.61 m
(3H, CH, Ph + NH), 8.54–8.32 m (1H, thiazole), 11.50
m (1H, NH). Mass spectrum, m/z (Irel, %): 408.2 (100)
[M + H]+. Found, %: C 59.07; H 5.07; N 23.94.
C20H21N7OS. Calculated, %: C 58.95; H 5.19; N 24.06.
4.2 Hz), 7.63–7.71 m (1H, CH, Ph), 8.38 s (1H, thia-
zole). Mass spectrum, m/z (Irel, %): 439.2 (100) [M +
H]+. Found, %: C 60.24; H 5.14; N 19.11. C22H23FN6OS.
Calculated, %: C 60.26; H 5.29; N 19.16.
1
Macrocycle 26а. Yield 63%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.21 s (1H, CH2,
piperidine), 1.51 s (1H, CH2, piperidine), 1.74 s (2H,
CH2, piperidine), 2.01 s (2H, CH2, piperidine), 2.34 s
(1H, CH2, piperidine), 2.37 s (3H, CH3-pyrimidine),
2.95 s (1H, CH2, piperidine), 3.77–3.93 m (3H, CH2 +
Macrocycle 23d was obtained as dihydrochloride
by treating with 16% HCl solution in dioxane. Yield
3
1
CH, piperidine), 4.37 d.d.d (2H, CH2NH, JHH = 6.8,
30%. H NMR spectrum (400 MHz, DMSO-d6), δ,
16.2, 22.5 Hz), 6.73 s (1H, CH, pyrimidine), 7.18 d.d.d
ppm: 2.13 m (2H, CH2, pyrrolidine), 2.42 s (3H, CH3-
pyrimidine), 2.52–2.69 m (2H, CH2, pyrrolidine), 3.29
s (1H, CH2, pyrrolidine), 3.49 s (1H, CH2, pyrrolidine),
4.54 m (4H, CH2 + CH2NH), 4.78 s (1H, CH2,
pyrrolidine), 6.87 s (1H, CH, pyrimidine), 7.28 t (1H,
CH, Ph, 3JHH = 9.3 Hz), 7.54 s (1H, NH), 7.82–8.10 m
(2H, CH, Ph), 8.38 s (1H, thiazole), 12.18 s (1H, NH).
Mass spectrum, m/z (Irel, %): 425.1 (100) [M + H]+.
Found, %: C 59.47; H 5.12; N 19.99. C21H21FN6OS.
Calculated, %: C 59.42; H 4.99; N 19.80.
3
(3H, CH, Ph, JHH = 7.2, 14.6, 22.1 Hz), 7.72 s (1H,
3
CH, Ph), 7.94 s (1H, thiazole), 8.47 t (1H, NH, JHH
=
6.6 Hz), 11.67 s (1H, NH). Mass spectrum, m/z (Irel,
%): 421.2 (100) [M + H]+. Found, %: C 62.68; H 5.44;
N 19.71. C22H24N6OS. Calculated, %: C 62.83; H 5.75;
N 19.98.
1
Macrocycle 26b. Yield 50%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.18 m (2H, CH2,
piperidine), 1.51 m (2H, CH2, piperidine), 1.67 m (2H,
CH2, piperidine), 1.98 m (1H, CH2, piperidine), 2.35 m
(1H, CH2, piperidine), 2.37 s (3H, CH3-pyrimidine),
2.85 s (1H, CH23, piperidine), 3.90 m (2H, CH2), 4.12 d
(2H, CH2NH, JHH = 12.5 Hz), 6.68 s (1H, CH,
pyrimidine), 7.27 d.d (2H, CH, Ph, 3JHH = 3.2, 7.5 Hz),
1
Macrocycle 23e. Yield 59%. H NMR spectrum
3
(400 MHz, DMSO-d6), δ, ppm: 1.63 d (1H, JHH
=
8.0 Hz, CH2, pyrrolidine), 1.70–1.83 m (1H, CH2,
3
pyrrolidine), 1.92 d (1H, CH2, pyrrolidine, JHH
=
3
2.1 Hz), 2.27 d.d (2H, CH2, pyrrolidine, JHH = 19.4,
9.8 Hz), 2.35 s (3H, CH3-pyrimidine), 3.00–3.12 m
(1H, CH2, pyrrolidine), 3.48 d (1H, CH, pyrrolidine,
3
7.72 t (1H, CH, Ph, JHH = 7.6 Hz), 7.79 s (1H,
thiazole), 8.23 t (1H, NH, 3JHH = 5.2 Hz), 11.73 s (1H,
NH). Mass spectrum, m/z (Irel, %): 422.1 (100) [M + H]+.
Found, %: C 59.72; H 5.44; N 23.10. C21H22N7OS.
Calculated, %: C 59.84; H 5.50; N 23.26.
3JHH = 13.8 Hz), 4.03 d.d (2H, CH2, JHH = 20.6,
3
10.7 Hz), 4.34 d.d (1H, CH2, 3JHH = 16.6, 5.3 Hz), 4.63
d.d (1H, CH2, 3JHH = 16.7, 7.1 Hz), 4.54 m (4H, CH2 +
CH2NH), 6.71 s (1H, CH, pyrimidine), 7.10 d (1H,
1
Macrocycle 26c. Yield 37%. H NMR spectrum
3
3
CH, Ph, JHH = 7.7 Hz), 7.29 d (1H, CH, Ph, JHH
=
(400 MHz, DMSO-d6), δ, ppm: 1.48 m (1H, CH2,
piperidine), 1.68 m (2H, CH2, piperidine), 1.82 m (1H,
CH2, piperidine), 2.20 s (1H, CH2, piperidine), 2.36–
2.38 m (3H, CH3-pyrimidine), 2.87 d (1H, CH2, pipe-
ridine, 3JHH = 11.0 Hz), 3.21–3.63 m (3H, CH2, piperi-
dine), 3.76–4.10 m (3H, CH2 + CH2NH), 4.31 m (1H,
7.9 Hz), 7.74 s (1H, CH, Ph), 8.43 s (1H, NH), 8.70 s
(1H, thiazole), 11.71 s (1H, NH). Mass spectrum, m/z
(Irel, %): 440.98 (100) [M + H]+. Found, %: C 57.16; H
4.95; N 19.18. C21H21ClN6OS. Calculated, %: C 57.20;
H 4.80; N 19.06.
3
CH2NH), 6.61 d (1H, CH, pyrimidine, JHH = 3.6 Hz),
1
Macrocycle 23f. Yield 49%. H NMR spectrum
3
6.83 d (0.5H, CH, Ph, JHH = 8.0 Hz), 7.07 d (0.5H,
(400 MHz, DMSO-d6), δ, ppm: 1.73–1.93 m (3H, CH2,
CH, Ph, 3JHH = 6.5 Hz), 7.47 s (0.5H, CH, Ph), 7.52 m
(0.5H, CH, Ph), 7.62–7.73 m (1H, CH, Ph), 8.21–8.39
m (1H, thiazole), 8.45 s (1H, NH), 11.34 s (1H, NH).
Mass spectrum, m/z (Irel, %): 422.2 (100) [M + H]+.
Found, %: C 59.72; H 5.58; N 23.18. C21H22N7OS.
Calculated, %: C 59.84; H 5.50; N 23.26.
3
pyrrolidine), 2.14 d.t (1H, CH2, pyrrolidine, JHH
=
19.5, 8.1 Hz), 2.33 s (3H, CH3-pyrimidine), 2.41–2.46
m (1H, CH2, pyrrolidine), 2.87–2.94 m (1H, CH2,
3
pyrrolidine, JHH = 2.1 Hz), 2.98 m (3H, CH3), 3.43 d
3
3
(1H, CH2, JHH = 13.2 Hz), 3.78 d (1H, CH2, JHH
=
3
13.2 Hz), 3,89 d.d (1H, CH, pyrrolidine, JHH = 4.4,
8.4 Hz), 4.43 d.d (2H, CH2NH, JHH = 15.5, 41.8 Hz),
1
3
Macrocycle 26d. Yield 42%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.10 d (1H, CH2, pipe-
ridine, JHH = 12.7 Hz), 1.46 d (2H, CH2, piperidine,
6.66 s (1H, CH, pyrimidine), 7.07 d.d (1H, CH, Ph,
3
3JHH = 8.4, 10.4 Hz), 7.20 d.d (1H, CH, Ph, 3JHH = 6.7,
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 87 No. 4 2017

DESIGN AND DEVELOPMENT OF MACROCYCLIZATION METHODS
729
3JHH = 31.4 Hz), 1.62–1.76 m (1H, CH2, piperidine),
1.99 d.d (1H, CH2, piperidine, JHH = 20.1, 7.0 Hz),
acid 29a–29c in 100 mL of THF was added 321 mg
(1.0 mmol) of TBTU and 0.17 mL (1.2 mmol) of
triethylamine. The reaction mixture was stirred for
16 h, and then concentrated in a vacuum. A saturated
solution of potassium carbonate (25 mL) was added to
the residue, and the mixture was extracted with
dichloromethane (2 × 25 mL). The organic layer was
separated, dried with sodium sulfate, and evaporated.
The desired product was isolated by chromatography
on a silica gel column (eluent ethyl acetate) and
crystallized from hexane.
3
2.18–2.30 m (2H, CH2, piperidine), 2.38 s (3H, CH3-
3
pyrimidine), 2.84 t (1H, CH2, piperidine, JHH
=
10.8 Hz), 3.82 d (1H, CH, piperidine, 3JHH = 13.6 Hz),
3.95 d (2H, CH2, JHH = 13.7 Hz), 4.33 d.d (1H,
3
3
CH2NH, JHH = 5.6, 16.4 Hz), 4.56 d.d (1H, CH2NH,
3JHH = 7.2, 16.4 Hz), 6.72 s (1H, CH pyrimidine), 6.98–
7.06 m (1H, CH, Ph), 7.13–7.21 m (1H, CH, Ph), 7.71
3
s (1H, CH, Ph), 8.18 d (1H, thiazole, JHH
=
3
7.4 Hz), 8.60 t (1H, NH, JHH = 6.1 Hz), 11.72 s (1H,
NH). Mass spectrum, m/z (Irel, %): 439.2 (100) [M + H]+.
Found, %: C 59.99; H 5.38; N 19.21. C22H23FN6OS.
Calculated, %: C 60.26; H 5.29; N 19.16.
1
Macrocycle 30а. Yield 30%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.8–2.4 m (3H, CH2,
pyrrolidine), 2.28 m (1H, CH2, pyrrolidine), 3.05 s
(3H, CH3-pyrimidine), 3.41 m (1H, CH2, pyrrolidine),
3.61 m (1H, CH2, pyrrolidine), 3.85 s (3H, CH3N),
4.45 m (2H, CH2), 4.71 m (1H, CH, pyrrolidine), 6.72
s (1H, CH, pyrimidine), 7.62 m (3H, CH, Ph), 8.17 s
(1H, thiazole), 11.75 br.s (1H, NH). Mass spectrum,
m/z (Irel, %): 501.2 (100) [M + H]+. Found, %: C 52.62;
H 5.00; N 16.58. C22H24N6O4S2. Calculated, %: C
52.78; H 4.83; N 16.79.
1
Macrocycle 26e. Yield 55%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.05 s (1H, CH2, pipe-
3
ridine ), 1.31 d (1H, CH2, piperidine, JHH = 12.0 Hz),
1.51 s (1H, CH2, piperidine), 1.68 d (1H, CH2, piperi-
dine, 3JHH = 12.2 Hz), 2.04 t (1H, CH2, piperidine, 3JHH
=
12.1 Hz), 2.28 d (2H, CH2, piperidine, 3JHH = 13.6 Hz),
2.38 s (3H, CH3-pyrimidine), 2.84 t (1H, CH2, piperi-
dine, 3JHH = 12.4 Hz), 3.86 d (1H, CH, piperidine, 3JHH
=
=
3
14.1 Hz), 3.98 s (1H, CH2), 4.08 d (1H, CH2, JHH
1
Macrocycle 30b. Yield 34%. H NMR spectrum
3
14.1 Hz), 4.31 d.d (1H, CH2NH, JHH = 5.3, 16.8 Hz),
(400 MHz, DMSO-d6), δ, ppm: 1.76 m (1H, CH2,
pyrrolidine), 2.04–2.20 m (4H, CH2, pyrrolidine), 2.41
s (3H, CH3-pyrimidine), 2.90 m (1H, CH2, pyrro-
lidine), 3.68 m (4H, CH2), 3.90 s (3H, CH3O), 4.59 m
(1H, CH, pyrrolidine), 6.75 s (1H, CH, pyrimidine),
7.18 m (1H, CH, Ph), 7.65–7.87 m (4H, CH, Ph +
thiazole), 11.76 br.s (1H, NH). Mass spectrum, m/z
(Irel, %): 501.4 (100) [M + H]+. Found, %: C 52.72; H
4.99; N 16.67. C22H24N6O4S2. Calculated, %: C 52.78;
H 4.83; N 16.79.
3
4.64 d.d (1H, CH2NH, JHH = 7.5, 16.9 Hz), 6.78 s
3
(1H, CH, pyrimidine), 7.18 d (1H, CH, Ph, JHH
=
3
7.8 Hz), 7.31 d (1H, CH, Ph, JHH = 8.0 Hz), 7.72 s
(1H, CH, Ph), 8.28 s (1H, thiazole), 8.71 s (1H, NH),
11.84 s (1H, NH). Mass spectrum, m/z (Irel, %): 455.0
(100) [M + H]+. Found, %: C 58.19; H 5.08; N 18.22.
C22H23ClN6OS. Calculated, %: C 58.08; H 5.10; N 18.47.
1
Macrocycle 26f. Yield 51%. H NMR spectrum
(400 MHz, DMSO-d6), δ, ppm: 1.24 d (1H, CH2, piperi-
3
dine, JHH = 23.9 Hz), 1.45 d (1H, CH2, piperidine,
1
Macrocycle 30c. Yield 30%. H NMR spectrum
3
3JHH = 23.7 Hz), 1.68 t (1H, CH2, piperidine, JHH
=
(400 MHz, DMSO-d6), δ, ppm: 1.72 m (1H, CH2,
pyrrolidine), 1.90 m (3H, CH2, pyrrolidine), 2.16–2.20
m (3H, CH2, pyrrolidine + CH2), 2.40 s (3H, CH3-
pyrimidine), 2.86 m (1H, CH2), 3.33 m (2H, CH2),
3.47 m (1H, CH2), 3.59 m (1H, CH2), 3.74 s (3H,
CH3O), 4.85 m (1H, CH, pyrrolidine), 6.89 s (1H, CH,
pyrimidine), 6.97 m (1H, CH, Ph), 7.37 m (1H, CH,
Ph), 7.75 m (2H, CH, Ph + thiazole), 11.76 br.s (1H,
NH). Mass spectrum, m/z (Irel, %): 515.4 (100) [M + H]+.
Found, %: C 53.72; H 5.16; N 16.43. C23H26N6O4S2.
Calculated, %: C 53.68; H 5.09; N 16.33.
15.1 Hz), 1.78–2.05 m (3H, CH2, piperidine), 2.37 s
(3H, CH3-pyrimidine), 2.39–2.44 m (1H, CH2, piperi-
dine), 2.84 t (1H, CH2, piperidine, 3JHH = 9.6 Hz), 3.00
3
s (3H, CH3), 3.50 d (1H, CH, piperidine, JHH
=
13.3 Hz), 3.62 d (1H, CH2, 3JHH = 14.5 Hz), 3.74 t (1H,
CH2, JHH = 12.9 Hz), 3.98 s (1H, CH2), 4.41–4.58 m
3
(2H, CH2NH), 6.72 s (1H, CH, pyrimidine), 7.03–7.12
m (1H, CH, Ph), 7.20–7.29 m (1H, CH, Ph), 7.73 s
3
(1H, CH, Ph), 8.10 d (1H, thiazole, JHH = 7.2 Hz),
11.60 s (1H, NH). Mass spectrum, m/z (Irel, %): 453.2
(100) [M + H]+. Found, %: C 61.12; H 5.58; N 18.42.
C23H25FN6OS. Calculated, %: C 61.04; H 5.57; N 18.57.
ACKNOWLEDGMENTS
The work was supported by the Ministry of
Education and Science of the Russian Federation
General procedure for the synthesis of macro-
cycles 30a–30c. To a solution of 0.8 mmol of amino
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 87 No. 4 2017

730
VASILEVICH et al.
p. 1641.
(contract no. 14.576.21.0019, unique identifier
RFMEFI57614X0019).
4. Mallinson, J. and Collins, I., Future Med. Chem., 2012,
vol. 4, no. 11, p. 1409. doi 10.4155/fmc.12.93
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