Diversity-oriented synthesis of amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-ones and evaluation of their influence on melanin synthesis in murine B16 cells

Article abstract of DOI:10.1515/hc-2017-0256

A diversity-oriented synthesis of amide-containing thieno[2,3-d]pyrimidin-4(3H)-ones is reported. All compounds were tested for their influence on melanin synthesis in murine B16 cells. The azepine fragment in thieno[2,3-d]pyrimidin-4(3H)-one skeleton significantly increases the melanin content.

Full text of DOI:10.1515/hc-2017-0256

Heterocycl. Commun. 2018; aop
Li Fei Nie, Guozheng Huang, Khurshed Bozorov, Jiangyu Zhao, Chao Niu,
Shamansur S. Sagdullaev and Haji A. Aisa*
Diversity-oriented synthesis of amide derivatives of tricyclic
thieno[2,3-d]pyrimidin-4(3H)-ones and evaluation of their
influence on melanin synthesis in murine B16 cells
https://doi.org/10.1515/hc-2017-0256
functional and physiological abnormalities in the
affected skin. So far, several theories have been gener-
ated to explain the pathogenesis of the vitiligo disease
[2]. It is generally believed that the disease results from
the destruction of melanocytes and obstruction of the
melanin synthesis [3].
Presently, there are several drugs used for the treat-
ment of vitiligo including topical corticosteroids, cal-
cineurin inhibitors, vitamin D3 analogs and psoralens
Received December 11, 2017; accepted December 18, 2017
Abstract: A diversity-oriented synthesis of amide-con-
taining thieno[2,3-d]pyrimidin-4(3H)-ones is reported.
All compounds were tested for their influence on melanin
synthesis in murine B16 cells. The azepine fragment in
thieno[2,3-d]pyrimidin-4(3H)-one skeleton significantly
increases the melanin content.
Keywords:melaninsynthesis;murineB16cells;thieno[2,3- [4]. Several psoralens (furocoumarins) have been clini-
d]pyrimidin-4(3H)-one; vitiligo.
cally applied, including 8-methoxypsoralen (8-MOP,
Figure 1) [5, 6], 5-methoxypsoralen [7] and 4,5,8-tri-
methylpsoralen [8, 9]. These compounds show strong
photosensitivity, which may be used for the treatment
of vitiligo with subsequent exposure to long-wave ultra-
violet (UV) radiation. Nowadays, this therapy is still the
Introduction
Vitiligo is a disease characterized by loss of pigment most successful approach for the treatment of vitiligo,
cells in the epidermis [1]. It can involve any part of although there are some undesired side effects such
the body where melanocytes reside and cause both as genetic mutation and risk of skin cancer [10, 11]. In
order to discover new active drug candidates, our group
has synthesized dozens of psoralen derivatives. In vitro
*Corresponding author: Haji A. Aisa, Key Laboratory of Plant
evaluation has indicated that some of them possess good
Resources and Chemistry in Arid Regions, State Key Laboratory
stimulation effects on tyrosinase and melanin synthesis
Basis of Xinjiang Indigenous Medicinal Plants Resource Utilization,
in murine B16 cells [12, 13].
Xinjiang Technical Institute of Physics and Chemistry, Chinese
Academy of Sciences, South Beijing Rd 40-1, Urumqi 830011,
P. R. China, e-mail: haji@ms.xjb.ac.cn
Thieno[2,3-d]pyrimidin-4(3H)-one is
a
common
building block for drugs with diverse pharmaceutical
activities [14–16]. It has been widely used for the prepa-
ration of new antibacterial [17, 18] and antitumor agents
[19], among other drugs. As can be seen from Figure 1,
the core of thieno[2,3-d]pyrimidin-4(3H)-one is similar to
that of psoralen, and we envisioned that derivatives of
thieno[2,3-d]pyrimidin-4(3H)-one might function simi-
larly to psoralen derivatives. Recently, 18 new sulfona-
mides derived from 6,7,8,9-tetrahydro-4H-pyrido[1,2-a]
thieno[2,3-d]pyrimidin-4-one have been synthesized by
our group. Several compounds exhibited more than 1.5-
fold potency as compared to 8-MOP [5]. In the present
study, inspired by these positive results, we synthesized
51 novel amide derivatives of tricyclic thieno[2,3-d]pyri-
midines and investigated their activities on melanin syn-
thesis in murine B16 cells.
Li Fei Nie, Guozheng Huang, Jiangyu Zhao and Chao Niu: Key
Laboratory of Plant Resources and Chemistry in Arid Regions,
State Key Laboratory Basis of Xinjiang Indigenous Medicinal Plants
Resource Utilization, Xinjiang Technical Institute of Physics and
Chemistry, Chinese Academy of Sciences, South Beijing Rd 40-1,
Urumqi 830011, P. R. China.
http://orcid.org/0000-0002-5730-9549 (G. Huang)
Khurshed Bozorov: Institute of the Chemistry of Plant Substances,
Academy of Sciences of Uzbekistan, MirzoUlugbek Str. 77, Tashkent
100170, Uzbekistan; and Key Laboratory of Plant Resources and
Chemistry in Arid Regions, State Key Laboratory Basis of Xinjiang
Indigenous Medicinal Plants Resource Utilization, Xinjiang Technical
Institute of Physics and Chemistry, Chinese Academy of Sciences,
South Beijing Rd 40-1, Urumqi 830011, P. R. China.
http://orcid.org/0000-0001-7449-0936 (K. Bozorov)
Shamansur S. Sagdullaev: Institute of the Chemistry of Plant
Substances, Academy of Sciences of Uzbekistan, MirzoUlugbek
Str. 77, Tashkent 100170, Uzbekistan
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2ꢀ
ꢀL. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-ones
Amide side-chain
Sulfonamide side-chain
O
O
N
N
O
O
O
e
M
e
M
H
C
n
)
(
2
S
O
S
N
N
n = 1–3
8-MOP
This work
Previous work
Figure 1ꢀStructures of 8-methoxypsolaren (8-MOP) and related derivatives of thieno[2,3-d]pyrimidin-4(3H)-one [5].
Results and discussion
n-Butylamine and 1-(4-aminophenyl)ethan-1-one were
selected to represent aliphatic and aromatic amines, respec-
tively. Using the coupling reagents EDCI and HOBt, the
yields were only moderate for both aliphatic amine (53%)
and aromatic amine (65%). HATU provided a good yield for
the reaction of aliphatic amine (88%), slightly better than
for aromatic amine (74%). The conversion of acid 6 to car-
boxylic chloride and then condensation with amine pro-
vided excellent yields with the aromatic amine (90%), much
better than for the aliphatic n-butylamine. Therefore, it was
decided to synthesize the target compounds 9a–e, 10a–e,
11a–e bearing the aliphatic amino moiety in the presence of
HATU and to prepare compounds 9f–q, 10f–q, 11f–q with
aromatic amine moiety from carboxylic acid chloride.
Synthesis
The synthetic route of the target compounds (Scheme 1)
follows our previous procedure [5]. An intermediate product
1 was prepared from diethyl oxalate via the Grignard reac-
tion. Application of the Gewald reaction to compound 1,
cyanoacetate and sulfur in the presence of triethylamine
at room temperature produced compound 2 in high yield.
Then, ethyl thieno[2,3-d]pyrimidine carboxylates 3–5 were
prepared through the condensation of compound 2 and
cyclic lactams in the presence of phosphorus oxychloride
in dichloromethane at 45°C. Hydrolysis of esters 3–5 was
completed in a mixture of water and methanol in the pres-
ence of lithium hydroxide to yield the acids 6–8. In order Melanin synthesis evaluation
to obtain the amidation products in good yields, the opti-
mization reactions with acid 6 under different conditions All the tricyclic thieno[2,3-d]pyrimidin-4(3H)-one deriva-
using a mixture of 1-ethyl-3-(3-dimethylaminopropyl)car- tives 9a–q, 10a–q, 11a–q were tested for their effect on
bodiimide and hydroxybenzotriazole (EDCI and HOBt), melanin synthesis in murine B16 cells, following a pre-
(1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b] viously published method [20]. As shown in Table 1, the
pyridinium 3-oxide hexafluorophosphate (HATU) and reference drug 8-MOP enhances the melanin synthesis by
oxalyl chloride as activating reagents were conducted. 126.1ꢀ ꢀ3.2% at 50 μm as compared to the blank control.
O
HN
(CH2)n
O
O
NC
EtO2C
CO2Et
NH2
MgBr
O
O
OEt
O
EtO
OEt
S8, EtOH, NEt3, rt
THF, –10°C
POCl3, DCM, 45°C
O
O
S
1
2
O
O
N
O
R
EtO2C
HO2C
(COCl)2, amine
3, 6, 9a-q, n = 1
LiOH
Et3N, DCM
N
N
N
4
,
7 10a q
, - , n = 2
(CH2)n
(CH2)n
(CH2)n
or HATU, amine
Et3N, DCM
5, 8,11a-q, n = 3
MeOH
S
S
S
N
N
9a-q, 10a-q, 11a-q
3
4
5
6 7 8
, ,
,
,
H
N
H
N
N
a
b
c
d
e
: R=
: R=
: R=
: R=
: R=
f: R=
l: R=
N
N
H
NH
H
Cl
H
H
N
H
N
H
N
H
H
N
N
N
j
g: R=
h: R=
i: R=
: R=
k: R=
Cl
O
O
Cl
H
H
N
H
H
N
N
N
N
NH
m
: R=
n
o
p
q
: R=
: R=
: R=
: R=
N
Br
Cl
Scheme 1ꢀSynthetic route to compounds 9a–q, 10a–q and 11a–q.
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L. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-onesꢂ ꢂ3
Table 1ꢀStimulation of melanin content of B16 cells by compounds 9a–q, 10a–q, 11a–q.
Compoundꢁ
Relative melainꢁ
contents (%)
Compoundꢁ
Relative melainꢁ
contents (%)
Compoundꢁ
Relative melain
contents (%)
9a
9b
9c
9d
9e
9f
9g
9h
9i
9j
9k
9l
9m
9n
9o
9p
9q
8-MOP
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
87.2ꢁ ꢁ2.5ꢃ
125.6ꢁ ꢁ3.4ꢃ
126.4ꢁ ꢁ5.5ꢃ
151.4ꢁ ꢁ6.4ꢃ
125.6ꢁ ꢁ3.4ꢃ
85.5ꢁ ꢁ6.9ꢃ
88.5ꢁ ꢁ7.8ꢃ
103.5ꢁ ꢁ5.2ꢃ
147.1ꢁ ꢁ2.6ꢃ
122.5ꢁ ꢁ7.1ꢃ
96.1ꢁ ꢁ8.4ꢃ
116.6ꢁ ꢁ6.8ꢃ
114.7ꢁ ꢁ4.9ꢃ
113.1ꢁ ꢁ5.6ꢃ
116.9ꢁ ꢁ7.1ꢃ
114.6ꢁ ꢁ7.6ꢃ
75.1ꢁ ꢁ4.6ꢃ
10a
10b
10c
10d
10e
10f
10g
10h
10i
10j
10k
10l
10m
10n
10o
10p
10q
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
109.1ꢁ ꢁ9.9ꢃ
105.9ꢁ ꢁ1.4ꢃ
133.2ꢁ ꢁ1.8ꢃ
174.5ꢁ ꢁ4.7ꢃ
159.6ꢁ ꢁ5.1ꢃ
151.9ꢁ ꢁ5.2ꢃ
103.0ꢁ ꢁ6.8ꢃ
227.6ꢁ ꢁ4.3ꢃ
112.5ꢁ ꢁ7.8ꢃ
126.4ꢁ ꢁ9.5ꢃ
187.9ꢁ ꢁ8.6ꢃ
155.8ꢁ ꢁ7.9ꢃ
201.2ꢁ ꢁ6.7ꢃ
154.4ꢁ ꢁ5.6ꢃ
130.7ꢁ ꢁ6.2ꢃ
154.4 8 .6ꢃ
95.4ꢁ ꢁ3.9ꢃ
11a
11b
11c
11d
11e
11f
11g
11h
11i
11j
11k
11l
11m
11n
11o
11p
11q
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
ꢃ
97.1ꢁ ꢁ3.2
155.8ꢁ ꢁ5.6
161.7ꢁ ꢁ7.1
111.5ꢁ ꢁ9.8
49.1ꢁ ꢁ7.6
143.9ꢁ ꢁ6.3
173.4ꢁ ꢁ10.8
82.9ꢁ ꢁ13.1
116.6ꢁ ꢁ3.7
92.5ꢁ ꢁ10.9
105.3ꢁ ꢁ6.1
147.0ꢁ ꢁ4.3
136.7ꢁ ꢁ8.2
234.5ꢁ ꢁ5.9
94.5ꢁ ꢁ10.7
150.4ꢁ ꢁ4.6
40.6ꢁ ꢁ11.4
126.1ꢁ ꢁ3.2
The data were recorded as the meanꢁ ꢁSD% of three experiments in duplicate and compared to the blank control.
The effectiveness of the synthetic compounds on melanin
Conclusion
synthesis varies from 40.6ꢀ ꢀ11.4 to 234.5ꢀ ꢀ5.9%. It can be
noticed that the five-membered ring compounds (9a–q,
New amide-substituted tricyclic thieno[2,3-d]pyrimidin-
4(3H)-one derivatives were synthesized and evaluated
for their effect on melanin synthesis in murine B16 cells.
Several synthetic compounds are more potent than the
standard drug 8-methoxypsolaren.
nꢀ=ꢀ1) showed decreased melanin synthesis, no matter
what kind of substituent group is present on the amide
moiety. Among these 17 compounds, only compounds
9d and 9i showed higher values than 8-MOP. Several
compounds fused with a seven-membered ring (11, nꢀ=ꢀ3)
possess better activities to promote melanin content.
Some six-membered derivatives (10, nꢀ=ꢀ2) show high
potency. Most of them (11 out of 17) increase the synthesis
of melanin. It can be concluded that the size of fused rings
of the synthetic compounds dramatically affects their
ability to influence melanin synthesis.
Experimental
Reagents and solvents were purchased from Sigma and used with-
out purification. Thin-layer chromatography (TLC) was carried out
on glass plates coated with silica gel (Qingdao Haiyang Chemical
Co., G60F-254); spots were visualized by illumination with UV light
(254 nm). The products were purified by column chromatography over
silica gel (Qingdao Haiyang Chemical Co., 200–300 mesh). Melting
points were determined on a Buchi B-540 apparatus and are uncor-
rected. Proton nuclear magnetic resonance (1H NMR), (400 MHz) and
carbon-13 nuclear magnetic resonance (13C NMR) (100 MHz) spectra
were recorded on a Varian NMR spectrometer in deuterated chloro-
form (CDCl3) using tetramethylsilane (TMS) as an internal standard.
High-resolution mass spectra (HRMS) were recorded on an AB SCIEX
QSTAR Elite quadrupole time-of-flight mass spectrometer.
Substituents on the amide moiety also contribute to
the biological effectiveness. For example, compounds
with n-octylamino group 9d, 10d, 11d exhibit an increased
potency in comparison to the derivatives substituted with
an n-butylamino group 9a, 10a, 11a. Compound 11n is the
lead candidate showing the highest potency on melanin
synthesis in murine B16 cells, with a value of 234.5ꢀ ꢀ5.9%,
which is 1.86-fold greater compared to that of 8-MOP.
Another hit compound is 10h which contains electron-
donating substituent (methoxy group) attached to posi-
tion 4 of the aromatic ring and increases the melanin
content by 227.6ꢀ ꢀ4.3%. Interestingly, activities of com-
pounds with a 2,6-dichlorophenyl fragment in amide
linker increase with the increase in size of the saturated
ring 9nꢀ<ꢀ10nꢀ<ꢀ11n.
General procedure for the synthesis of compounds 9a–q,
10a–q and 11a–q
Method A To a solution of 2-methyl-4-oxo-thieno[2,3-d]pyrimi-
dine-3-carboxylic acid (6–8, 1.0 mmol) and HATU (1.2 mmol) in
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4ꢀ ꢀL. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-ones
dichloromethane (20 mL) was added N,N-diisopropylethylamine
(DIPEA, 0.27 mL, 1.5 mmol) and an aliphatic amine (see Scheme 1,
1.5 mmol) dropwise at 0–5°C. The mixture was stirred at room tem-
perature overnight and then concentrated under reduced pressure.
The residue was extracted with dichloromethane (2ꢀ×ꢀ30 mL). The
extract was washed with brine, dried over anhydrous MgSO4, filtered
and concentrated under reduced pressure. The residue was purified
by silica gel chromatography to produce the pure compounds 9a–e,
10a–e, 11a–e.
(t, Jꢀ=ꢀ7 Hz, 2H), 3.98–3.87 (m, 1H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.86 (s, 3H),
2.38–2.30 (m, 2H), 2.06–1.99 (m, 2H), 1.82–1.73 (m, 2H), 1.58–1.17 (m,
6H); 13C NMR: δ 163.4, 162.3, 159.2, 158.5, 145.6, 126.6, 118.7, 48.5, 47.4,
32.9, 32.0, 25.8, 25.0, 19.6, 17.5. HRMS (ESI). Calcd for C17H21N3O2S [M–
H]− m/z 331.1348. Found: m/z 331.1354.
2-Methyl-4-oxo-N-(o-tolyl)-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9f)ꢁYield 87%; white
1
solid; mp 213–215°C; H NMR: δ 12.45 (s, 1H), 7.68 (d, Jꢀ=ꢀ8 Hz, 1H),
7.25–7.18 (m, 2H), 7.12–7.05 (m, 1H), 4.15 (t, Jꢀ=ꢀ7 Hz, 2H), 3.10 (t, Jꢀ=ꢀ7 Hz,
2H), 2.89 (s, 3H), 2.42–2.35 (m, 2H), 2.35 (s, 3H); 13C NMR: δ 162.7, 161.7,
159.0, 158.4, 147.1, 135.9, 132.4, 129.4, 126.1, 120.5, 118.7, 45.2, 32.8, 23.6,
19.5, 17.7. HRMS (ESI). Calcd for C18H17N3O2S [M–H]−: m/z 339.1049.
Found: m/z 339.1041.
Method B A solution of 2-methyl-4-oxo-thieno[2,3-d]pyrimidine-3-car-
boxylic acid (6–8, 1.0 mmol) in dichloromethane (10 mL) was treated
with oxalyl chloride (3 mmol) and the mixture was stirred at room tem-
perature for 3 h. The solvent and the excess of oxalyl chloride were
removed under reduced pressure and the residue was treated with an
aromatic amine (see Scheme 1, 1.1 mmol) and triethylamine (1.2 mmol).
The mixture was stirred at room temperature for 8 h, then washed with
water and concentrated under reduced pressure. The residue was puri-
fied by silica gel chromatography and eluted with petroleum ether/
ethyl acetate to give the pure compounds 9f–q, 10f–q, 11f–q.
2-Methyl-4-oxo-N-(o-tolyl)-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9g)ꢁYield 79%; white
solid; mp 257–259°C; 1H NMR: δ 12.77 (s, 1H), 7.67 (d, Jꢀ=ꢀ8 Hz, 2H), 7.14
(d, Jꢀ=ꢀ8 Hz, 2H), 4.25 (t, Jꢀ=ꢀ7 Hz, 2H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.93 (s, 3H),
2.39–2.30 (m, 2H), 2.33 (s, 3H); 13C NMR: δ 163.4, 161.0, 159.3, 159.0, 147.5,
136.6, 133.1, 129.3, 126.4, 120.3, 118.2, 47.5, 31.9, 20.9, 19.5, 18.0. HRMS
(ESI). Calcd for C18H17N3O2S [M–H]−: m/z 339.1048. Found: m/z 339.1041.
N-Butyl-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9a)ꢁYield 81%; light
1
yellow solid; mp 119–121°C; H NMR: δ 10.40 (d, Jꢀ=ꢀ3.8 Hz, 1H), 4.23
N-(4-Methoxyphenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(t, Jꢀ=ꢀ7 Hz, 2H), 3.41 (t, Jꢀ=ꢀ7 Hz, 2H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.87 (s, 3H),
2.38–2.30 (m, 2H), 1.69–1.59 (m, 2H), 1.49–1.39 (m, 2H), 0.96 (t, Jꢀ=ꢀ7 Hz,
3H); 13C NMR: δ 163.5, 163.3, 159.2, 158.6, 145.8, 126.3, 118.6, 47.4, 39.5,
32.0, 31.5, 20.4, 19.6, 17.5, 13.9. HRMS (ESI). Calcd for C15H19N3O2S
[M–H]−: m/z 305.1193. Found: m/z 305.1198.
1
(9h)ꢁYield 76%; light yellow solid; mp 201–203°C; H NMR: δ 12.70
(s, 1H), 7.69 (d, Jꢀ=ꢀ9 Hz, 2H), 6.88 (d, Jꢀ=ꢀ9 Hz, 2H), 4.24 (t, Jꢀ=ꢀ7 Hz,
2H), 3.81 (s, 3H), 3.21 (t, Jꢀ=ꢀ8 Hz, 2H), 2.92 (s, 3H), 2.39–2.28 (m, 2H);
13C NMR: δ 163.3, 160.9, 159.4, 158.9, 156.0, 147.4, 132.4, 126.3, 121.8,
118.3, 114.0, 55.5, 47.5, 31.9, 19.5, 17.9. HRMS (ESI). Calcd for C18H17N3O3S
[M–H]−: m/z 355.0999. Found: m/z 355.0991.
N-(sec-Butyl)-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9b)ꢁYield 89%; light
1
yellow solid; mp 113–115°C; H NMR: δ 10.21 (d, Jꢀ=ꢀ4.3 Hz, 1H), 4.23
N-(2,6-Dimethylphenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide (9i)ꢁYield
(t, Jꢀ=ꢀ7 Hz, 2H), 4.09–4.01 (m, 1H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.86 (s, 3H),
2.38–2.30 (m, 2H), 1.72–1.53 (m, 2H), 1.25 (d, Jꢀ=ꢀ7 Hz, 3H), 0.98 (t,
Jꢀ=ꢀ7 Hz, 3H); 13C NMR: δ 163.4, 162.6, 159.2, 158.5, 145.5, 126.7, 118.6,
46.8, 38.7, 32.0, 29.5, 20.1, 19.6, 17.5, 10.6. HRMS (ESI). Calcd for
C15H19N3O2S [M–H]−: m/z 305.1192. Found: m/z 305.1198.
1
86%; light yellow solid; mp 215–217°C; H NMR: δ 12.02 (s, 1H), 7.14–
7.07 (m, 3H), 4.24 (t, Jꢀ=ꢀ7 Hz, 2H), 3.22 (t, Jꢀ=ꢀ8.0 Hz, 2H), 2.91 (s, 3H),
2.39–2.32 (m, 2H), 2.31 (s, 6H); 13C NMR: δ 163.5, 161.5, 159.3, 158.8, 147.5,
135.3, 135.1, 127.9, 126.5, 125.8, 118.6, 47.4, 32.0, 19.6, 18.7, 17.8. HRMS
(ESI). Calcd for C19H19N3O2S [M–H]−: m/z 353.1191. Found: m/z 353.1198.
N,N-Diisopropyl-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-
a]thieno[2,3-d]pyrimidine-3-carboxamide (9c)ꢁYield 82%; white
solid; mp 207–209°C; 1H NMR: δ 4.16 (t, Jꢀ=ꢀ7 Hz, 2H), 3.79–3.72 (m, 1H),
3.57–3.50 (m, 1H), 3.16 (t, Jꢀ=ꢀ8 Hz, 2H), 2.44 (s, 3H), 2.28 (m, 2H), 1.67
(d, Jꢀ=ꢀ7 Hz, 3H), 1.58 (d, Jꢀ=ꢀ7 Hz, 3H), 1.14 (t, Jꢀ=ꢀ7 Hz, 6H); 13C NMR: δ
164.8, 163.6, 159.9, 156.2, 131.5, 130.8, 120.0, 51.2, 46.5, 46.0, 32.2, 21.3,
20.8, 20.2, 19.9, 19.7, 13.7. HRMS (ESI). Calcd for C17H23N3O2S [M–H]−:
m/z 333.1507. Found: m/z 333.1511.
N-(4-Acetylphenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-
a]thieno[2,3-d]pyrimidine-3-carboxamide (9j)ꢁYield 89%; light
1
yellow solid; mp 189–191°C; H NMR: δ 13.32 (s, 1H), 7.94 (d, Jꢀ=ꢀ9 Hz,
2H), 7.88 (d, Jꢀ=ꢀ9 Hz, 2H), 4.26 (t, Jꢀ=ꢀ7 Hz, 2H), 3.22 (t, Jꢀ=ꢀ8 Hz, 2H),
2.94 (s, 3H), 2.58 (s, 3H), 2.41–2.32 (m, 2H); 13C NMR: δ 197.0, 163.5, 161.3,
159.5, 159.0, 148.9, 143.9, 132.2, 129.5, 125.8, 119.4, 118.0, 47.6, 31.9, 26.4,
19.5, 18.1. HRMS (ESI). Calcd for C19H17N3O3S [M–H]−: m/z 367.0984.
Found: m/z 367.0991.
2-Methyl-N-octyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9d)ꢁYield 76%; white
N-(4-Chlorophenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-
a]thieno[2,3-d]pyrimidine-3-carboxamide (9k)ꢁYield 82%; white
solid; mp 252–254°C; 1H NMR: δ 13.05 (s, 1H), 7.79–7.71 (m, 2H), 7.32–7.27
(m, 2H), 4.26 (t, Jꢀ=ꢀ7 Hz, 2H), 3.23 (t, Jꢀ=ꢀ8 Hz, 2H), 2.93 (s, 3H), 2.41–2.33
(m, 2H); 13C NMR: δ163.6, 161.1, 159.4, 159.0, 148.3, 137.9, 128.7, 128.4, 126.0,
121.4, 118.1, 47.5, 31.9, 19.6, 18.0. HRMS (ESI). Calcd for C17H14ClN3O2S
[M–H]−: m/z 359.0486. Found: m/z 359.0495.
1
solid; mp 102–104°C; H NMR: δ 10.40 (d, Jꢀ=ꢀ3.5 Hz, 1H), 4.23 (t,
Jꢀ=ꢀ7 Hz, 2H), 3.43–3.36 (m, 2H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.87 (s, 3H), 2.38–
2.30 (m, 2H), 1.68–1.61 (m, 2H), 1.43–1.23 (m, 10H), 0.87 (t, Jꢀ=ꢀ7 Hz,
3H); 13C NMR: δ 163.5, 163.2, 159.2, 158.6, 145.8, 126.4, 118.6, 47.3, 39.7,
32.0, 31.8, 29.4, 29.3, 29.2, 27.2, 22.7, 19.6, 17.5, 14.1. HRMS (ESI). Calcd
for C19H27N3O2S [M–H]−: m/z 361.1831. Found: m/z 361.1824.
N-Cyclohexyl-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9e)ꢁYield 87%; light
N-(2-Chlorophenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
1
yellow solid; mp 186–188°C; H NMR: δ 10.27 (d, Jꢀ=ꢀ7 Hz, 1H), 4.23
(9l)ꢁYield 84%; light yellow solid; mp 220–222°C; H NMR: δ 12.03
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ꢁꢀꢀꢀ
L. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-onesꢂ ꢂ5
N-(sec-Butyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-
a]thieno[2,3-d]pyrimidine-3-carboxamide (10b)ꢁYield 86%;
(s, 1H), 8.11 (dd, Jꢀ=ꢀ8 Hz, 0.9 Hz, 1H), 7.41 (dd, Jꢀ=ꢀ8 Hz, 1.3 Hz, 1H),
7.32–7.27 (m, 1H), 7.09 (td, Jꢀ=ꢀ8 Hz, 1.5 Hz, 1H), 4.26 (t, Jꢀ=ꢀ7 Hz, 2H), 3.22
(t, Jꢀ=ꢀ8 Hz, 2H), 2.88 (s, 3H), 2.39–2.31 (m, 2H); 13C NMR: δ 163.3, 161.7,
159.6, 158.3, 147.4, 135.4, 129.5, 127.0, 126.7, 126.0, 125.5, 125.4, 118.5,
47.4, 32.0, 19.6, 17.7. HRMS (ESI). Calcd for C17H14ClN3O2S [M–H]−: m/z
359.0483. Found: m/z 359.0495.
1
light yellow solid; mp 109–111°C; H NMR: δ 10.19 (d, Jꢀ=ꢀ6 Hz, 1H),
4.07 (t, Jꢀ=ꢀ6 Hz, 2H), 4.08–4.02 (m, 1H), 2.99 (t, Jꢀ=ꢀ7 Hz, 2H), 2.86
(s, 3H), 2.06–2.01 (m, 2H), 1.97–1.91 (m, 2H), 1.70–1.56 (m, 2H), 1.25
(d, Jꢀ=ꢀ7 Hz, 3H), 0.98 (t, Jꢀ=ꢀ7 Hz, 3H); 13C NMR: δ 162.7, 161.6, 159.6,
154.8, 145.1, 126.8, 118.5, 46.7, 43.1, 31.5, 29.5, 22.1, 20.1, 19.0, 17.4, 10.6.
HRMS (ESI). Calcd for C16H21N3O2S [M–H]−: m/z 319.1350. Found: m/z
319.1354.
N-(4-Bromophenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(9m)ꢁYield 90%; light yellow solid; mp 224–226°C; 1H NMR: δ 13.06
(s, 1H), 7.71 (d, Jꢀ=ꢀ8 Hz, 2H), 7.44 (d, Jꢀ=ꢀ8 Hz, 2H), 4.27 (t, Jꢀ=ꢀ7 Hz, 2H),
3.23 (t, Jꢀ=ꢀ8 Hz, 2H), 2.94 (s, 3H), 2.42–2.32 (m, 2H); 13C NMR: δ 163.6,
161.1, 159.4, 159.1, 148.4, 138.4, 131.7, 126.0, 121.8, 118.1, 116.1, 47.5, 31.9,
19.6, 18.0. HRMS (ESI). Calcd for C17H14BrN3O2S [M–H]−: m/z 402.9996.
Found: m/z 402.9990.
N,N-Diisopropyl-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido-
[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide (10c)ꢁYield 89%;
light yellow solid; mp 187–189°C; 1H NMR: δ 4.13–4.04 (m, 1H), 3.98–
3.91 (m, 1H), 3.79–3.72 (m, 1H), 3.57–3.49 (m, 1H), 2.96 (t, Jꢀ=ꢀ6 Hz, 2H),
2.44 (s, 3H), 2.01–1.89 (m, 4H), 1.68 (d, Jꢀ=ꢀ7 Hz, 3H), 1.58 (d, Jꢀ=ꢀ7 Hz,
3H), 1.13 (dd, Jꢀ=ꢀ7 Hz, 4 Hz, 6H); 13C NMR: δ 165.0, 162.1, 157.2, 155.3,
131.2, 130.9, 119.7, 51.2, 45.9, 42.1, 31.6, 22.0, 21.3, 20.8, 20.2, 19.9, 19.2,
13.7. HRMS (ESI). Calcd for C18H25N3O2S [M–H]− m/z 347.1661. Found:
m/z 347.1667.
N-(2,6-Dichlorophenyl)-2-methyl-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(9n)ꢁYield 88%; white solid; mp 267–269°C; 1H NMR: δ 12.27 (s, 1H),
8.09 (d, Jꢀ=ꢀ8 Hz, 1H), 7.42 (d, Jꢀ=ꢀ2.4 Hz, 1H), 7.29–7.22 (m, 2H), 4.24
(t, Jꢀ=ꢀ7 Hz, 2H), 3.23 (t, Jꢀ=ꢀ8 Hz, 2H), 2.89 (s, 3H), 2.40–2.31 (m, 2H);
13C NMR: δ 163.4, 161.7, 159.65, 158.5, 148.1, 134.3, 129.9, 129.2, 127.3,
127.2, 126.1, 125.6, 118.4, 47.4, 32.0, 19.6, 17.8. HRMS (ESI). Calcd for
C17H13Cl2N3O2S [M–H]−: m/z 393.0101. Found: m/z 393.0106.
2-Methyl-N-octyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (10d)ꢁYield 81%; light
yellow solid; mp 103–105°C; 1H NMR: δ 10.42 (s, 1H), 4.06 (t, Jꢀ=ꢀ6 Hz,
2H), 3.40 (dd, Jꢀ=ꢀ12 Hz, 7 Hz, 2H), 3.00 (t, Jꢀ=ꢀ7 Hz, 2H), 2.86 (s, 3H),
2.06–2.02 (m, 2H), 1.98–1.90 (m, 2H), 1.68–1.60 (m, 2H), 1.44–1.25 (m,
10H), 0.88 (t, Jꢀ=ꢀ7 Hz, 3H); 13C NMR: δ 163.4, 161.7, 159.6, 154.7, 145.4,
126.5, 118.5, 43.1, 39.7, 31.8, 31.5, 29.4, 29.3, 29.2, 27.2, 22.7, 22.2, 19.0,
17.4, 14.1. HRMS (ESI). Calcd for C20H29N3O2S [M–H]−: m/z 375.1986.
Found: m/z 375.1980.
N-Benzyl-2-methyl-4-oxo-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9o)ꢁYield 73%; light
yellow solid; mp 142–144°C; 1H NMR: δ 10.96 (d, Jꢀ=ꢀ1.1 Hz, 1H), 7.40 (d,
Jꢀ=ꢀ7 Hz, 2H), 7.32 (t, Jꢀ=ꢀ7 Hz, 2H), 7.23 (t, Jꢀ=ꢀ7 Hz, 1H), 4.63 (d, Jꢀ=ꢀ4 Hz,
2H), 4.19 (t, Jꢀ=ꢀ7 Hz, 2H), 3.18 (t, Jꢀ=ꢀ8 Hz, 2H), 2.88 (s, 3H), 2.36–2.26
(m, 2H); 13C NMR: δ 163.5, 163.3, 159.3, 158.5, 146.4, 139.0, 128.5, 127.7,
126.9, 126.0, 118.6, 47.4, 43.4, 32.0, 19.6, 17.6. HRMS (ESI). Calcd for
C18H17N3O2S [M–H]−: m/z 339.1035. Found: m/z 339.1041.
N-Cyclohexyl-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-
a]thieno[2,3-d]pyrimidine-3-carboxamide (10e)ꢁYield 77%;
1
white solid; mp 195–197°C; H NMR: δ 10.23 (d, Jꢀ=ꢀ7 Hz, 1H), 4.07 (t,
Jꢀ=ꢀ6 Hz, 2H), 3.97 –3.90 (m, 1H), 3.00 (t, Jꢀ=ꢀ7 Hz, 2H), 2.85 (s, 3H),
2.06 –1.98 (m, 4H), 1.98–1.91 (m, 2H), 1.81–1.74 (m, 2H), 1.48–1.20 (m,
6H); 13C NMR: δ 162.4, 161.4, 159.5, 154.8, 145.2, 126.8, 118.5, 48.4, 43.1,
32.9, 31.4, 25.8, 25.0, 22.1, 19.0, 17.4. HRMS (ESI). Calcd for C18H23N3O2S
[M–H]−: m/z 345.1517. Found: m/z 345.1511.
2-Methyl-4-oxo-N-(pyridin-2-yl)-4,6,7,8-tetrahydropyrrolo[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (9p)ꢁYield 76%; light
yellow solid; mp 218–220°C; 1H NMR: δ 13.40 (s, 1H), 8.45 (dd, Jꢀ=ꢀ5 Hz,
1 Hz, 1H), 8.40 (d, Jꢀ=ꢀ8 Hz, 1H), 7.77–7.70 (m, 1H), 7.06–7.01 (m, 1H), 4.33
(t, Jꢀ=ꢀ7 Hz, 2H), 3.22 (t, Jꢀ=ꢀ8 Hz, 2H), 2.93 (s, 3H), 2.40–2.31 (m, 2H); 13C
NMR: δ 163.5, 161.6, 159.7, 158.8, 152.6, 148.4, 147.9, 138.4, 126.0, 119.1,
118.3, 115.0, 47.7, 32.0, 19.6, 17.9. HRMS (ESI). Calcd for C16H14N4O2S
[M–H]−: m/z 326.0833. Found: m/z 326.0837.
2-Methyl-4-oxo-N-(o-tolyl)-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (10f)ꢁYield 80%; light
1
yellow solid; mp 155–157°C; H NMR: δ 12.18 (s, 1H), 7.78 (d, Jꢀ=ꢀ8 Hz,
1H), 7.25–7.18 (m, 2H), 7.12–7.05 (m, 1H), 4.09 (t, Jꢀ=ꢀ6 Hz, 2H), 3.02 (t,
Jꢀ=ꢀ7 Hz, 2H), 2.91 (s, 3H), 2.38 (s, 3H), 2.07–2.01 (m, 2H), 1.99–1.91 (m,
2H). 13C NMR: δ 161.7, 161.6, 159.8, 154.9, 147.4, 136.6, 131.6, 130.5, 126.2,
126.9, 125.1, 124.9, 118.3, 43.2, 31.5, 22.1, 19.0, 18.5, 17.9. HRMS (ESI).
Calcd for C19H19N3O2S [M–H]−: m/z 353.1191. Found: m/z 353.1198.
2-Methyl-N-(4-methylpyridin-2-yl)-4-oxo-4,6,7,8-tetrahydro-
pyrrolo[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
(9q)ꢁYield 92%; light yellow solid; mp 166–168°C; H NMR: δ 13.32
(s, 1H), 8.21 (d, Jꢀ=ꢀ5 Hz, 1H), 8.18 (s, 1H), 6.81 (d, Jꢀ=ꢀ5 Hz, 1H), 4.27
(t, Jꢀ=ꢀ7 Hz, 2H), 3.16 (t, Jꢀ=ꢀ7 Hz, 2H), 2.92 (s, 3H), 2.23–2.11 (m, 2H);
13C NMR: δ 162.3, 161.4, 159.8, 156.2, 152.8, 148.6, 147.8, 146.9, 126.3,
119.7, 118.2, 115.0, 46.3, 31.9, 22.4, 19.1, 18.0. HRMS (ESI). Calcd for
C17H16N4O2S [M–H]−: m/z 326.0843. Found: m/z 340.0894.
2-Methyl-4-oxo-N-(p-tolyl)-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (10g)ꢁYield 75%; light
yellow solid, mp 226–228°C; 1H NMR: δ 12.81 (s, 1H), 7.67 (d, Jꢀ=ꢀ8 Hz,
2H), 7.14 (d, Jꢀ=ꢀ8 Hz, 2H), 4.09 (t, Jꢀ=ꢀ6 Hz, 2H), 3.01 (t, Jꢀ=ꢀ7 Hz, 2H), 2.92
(s, 3H), 2.33 (s, 3H), 2.07–2.01 (m, 2H), 1.98–1.90 (m, 2H); 13C NMR: δ
161.6, 161.1, 160.1, 154.9, 147.2, 136.7, 133.1, 129.3, 126.5, 120.3, 118.1, 43.4,
31.5, 22.2, 20.9, 19.0, 17.9. HRMS (ESI). Calcd for C19H19N3O2S [M–H]−:
m/z 353.1193. Found: m/z 353.1198.
N-Butyl-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]-
thieno[2,3-d]pyrimidine-3-carboxamide (10a)ꢁYield 79%; white
solid; mp 209–211°C; 1H NMR: δ 10.40 (d, Jꢀ=ꢀ4 Hz, 1H), 4.23 (t, Jꢀ=ꢀ7 Hz,
2H), 3.41 (t, Jꢀ=ꢀ7 Hz, 2H), 3.20 (t, Jꢀ=ꢀ8 Hz, 2H), 2.87 (s, 3H), 2.12–2.06
(m, 2H), 1.97–1.91 (m, 2H)., 1.67–1.57 (m, 2H), 1.48–1.38 (m, 2H), 0.97 (t,
Jꢀ=ꢀ7.3 Hz, 3H); 13C NMR: δ 163.4, 163.3, 159.2, 158.5, 145.8, 126.3, 118.6,
47.3, 39.4, 31.9, 31.4, 21.2, 20.4, 19.6, 17.5, 13.9. HRMS (ESI). Calcd for
C16H21N3O2S [M–H]−: m/z 319.1350. Found: m/z 319.1354.
N-(4-Methoxyphenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(10h)ꢁYield 86%; light yellow solid; mp 201–203°C; 1H NMR: δ 12.74
Brought to you by | Göteborg University - University of Gothenburg
Authenticated
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ꢀꢀꢀꢁ
6ꢀ ꢀL. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-ones
N-Benzyl-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]
thieno[2,3-d]pyrimidine-3-carboxamide (10o)ꢁYield 82%; white
solid; mp 113–115°C; 1H NMR: δ 10.90 (s, 1H), 7.41 (d, Jꢀ=ꢀ7 Hz, 2H), 7.32
(t, Jꢀ=ꢀ7 Hz, 2H), 7.24 (t, Jꢀ=ꢀ7 Hz, 1H), 4.64 (d, Jꢀ=ꢀ5 Hz, 2H), 4.03 (t,
Jꢀ=ꢀ6 Hz, 2H), 3.00 (t, Jꢀ=ꢀ6 Hz, 2H), 2.87 (s, 3H), 2.04–1.97 (m, 2H), 1.97–
1.90 (m, 2H); 13C NMR: δ 163.4, 161.4, 159.6, 154.9, 146.0, 139.0, 128.4,
127.7, 126.9, 126.1, 118.4, 43.5, 43.2, 31.4, 22.1, 19.0, 17.5. HRMS (ESI).
Calcd for C19H19N3O2S [M–H]−: m/z 353.1203. Found: m/z 353.1198.
(s, 1H), 7.70 (d, Jꢀ=ꢀ9 Hz, 2H), 6.89 (d, Jꢀ=ꢀ9 Hz, 2H), 4.10 (t, Jꢀ=ꢀ6 Hz,
2H), 3.81 (s, 3H), 3.03 (t, Jꢀ=ꢀ7 Hz, 2H), 2.92 (s, 3H), 2.08–2.02 (m, 2H),
2.00–1.92 (m, 2H); 13C NMR: δ 161.4, 161.0, 160.0, 155.9, 154.9, 147.1,
132.4, 126.4, 121.8, 118.1, 114.0, 55.5, 43.4, 31.4, 22.1, 19.0, 17.8. HRMS
(ESI). Calcd for C19H19N3O3S [M–H]−: m/z 369.1142. Found: m/z 369.1147.
N-(2,6-Dimethylphenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
(10i)ꢁYield 83%; white solid; mp 186–188°C; H NMR: δ 12.11 (s,
2-Methyl-4-oxo-N-(pyridin-2-yl)-6,7,8,9-tetrahydro-4H-pyrido-
[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide (10p)ꢁYield 75%;
white solid; mp 181–183°C; 1H NMR: δ 13.29 (s, 1H), 8.31 (dd, Jꢀ=ꢀ5 Hz,
1.2 Hz, 1H), 8.27 (d, Jꢀ=ꢀ8 Hz, 1H), 7.67–7.59 (m, 1H), 7.11–7.05 (m, 1H),
4.31 (t, Jꢀ=ꢀ7 Hz, 2H), 3.13 (t, Jꢀ=ꢀ8 Hz, 2H), 2.94 (s, 3H), 2.08–2.04
(m, 2H), 1.97–1.91 (m, 2H); 13C NMR: δ 162.7, 161.9, 159.8, 156.7, 152.4,
149.2, 147.9, 138.9, 126.2, 119.8, 118.6, 115.7, 47.7, 32.0, 19.6, 17.9. HRMS
(ESI). Calcd for C17H16N4O2S [M–H]−: m/z 340.0988. Found: m/z
340.0994.
1H), 7.12–7.04 (m, 3H), 4.08 (t, Jꢀ=ꢀ6 Hz, 2H), 3.03 (t, Jꢀ=ꢀ6 Hz, 2H), 2.91
(s, 3H), 2.31 (s, 6H), 2.08–2.01 (m, 2H), 2.00–1.92 (m, 2H); 13C NMR: δ
161.8, 161.6, 159.9, 154.9, 147.3, 135.3, 135.2, 127.9, 126.5, 125.8, 118.4, 43.3,
31.5, 22.1, 19.0, 18.7, 17.7. HRMS (ESI). Calcd for C20H21N3O2S [M–H]−: m/z
367.1360. Found: m/z 367.1354.
N-(4-Acetylphenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(10j)ꢁYield 74%; white solid; mp 230–232°C; 1H NMR: δ 13.35 (s, 1H),
7.95 (d, Jꢀ=ꢀ9 Hz, 2H), 7.89 (d, Jꢀ=ꢀ9 Hz, 2H), 4.31 (t, Jꢀ=ꢀ7 Hz, 2H), 3.25 (t,
Jꢀ=ꢀ8 Hz, 2H), 2.91 (s, 3H), 2.62 (s, 3H), 2.45–2.37 (m, 2H). 2.05–1.93 (m,
2H); 13C NMR: δ 197.1, 162.1, 160.2, 159.8, 159.6, 149.3, 143.8, 132.3, 129.5,
126.0, 119.5, 118.0, 45.6, 31.7, 26.6, 22.7, 19.3, 18.1. HRMS (ESI). Calcd for
C20H19N3O3S [M–H]−: m/z 381.1154. Found: m/z 381.1147.
2-Methyl-N-(4-methylpyridin-2-yl)-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
(10q)ꢁYield 84%; white solid; mp 190–192°C; H NMR: δ 13.29 (s,
1H), 8.28 (d, Jꢀ=ꢀ5 Hz, 1H), 8.24 (s, 1H), 6.85 (d, Jꢀ=ꢀ5 Hz, 1H), 4.19 (t,
Jꢀ=ꢀ6 Hz, 2H), 3.01 (t, Jꢀ=ꢀ6 Hz, 2H), 2.93 (s, 3H), 2.39 (s, 3H), 2.06–1.99
(m, 2H), 1.98–1.91 (m, 2H); 13C NMR: δ 161.7, 161.6, 159.9, 155.2, 152.8,
149.4, 148.0, 147.9, 126.2, 120.4, 118.1, 115.3, 43.4, 31.5, 22.1, 21.4, 19.0,
17.9. HRMS (ESI). Calcd for C18H18N4O2S [M–H]−: m/z 354.1154. Found:
m/z 354.1150.
N-(4-Chlorophenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
(10k)ꢁYield 83%; light yellow solid; mp 238–240°C; 1H NMR: δ 13.15
(s, 1H), 7.76 (d, Jꢀ=ꢀ9 Hz, 2H), 7.30 (d, Jꢀ=ꢀ9 Hz, 2H), 4.12 (t, Jꢀ=ꢀ6 Hz, 2H),
3.03 (t, Jꢀ=ꢀ6 Hz, 2H), 2.93 (s, 3H), 2.11–2.03 (m, 2H), 2.01–1.93 (m, 2H); 13C
NMR: δ 161.8, 161.2, 160.2, 154.9, 148.1, 137.9, 128.7, 128.3, 126.0, 121.4,
117.9, 43.5, 31.5, 22.1, 19.0, 18.0. HRMS (ESI). Calcd for C18H16ClN3O2S
[M–H]−: m/z 373.0646. Found: m/z 373.0652.
N-Butyl-2-methyl-4-oxo-4,6,7,8,9,10-hexahydrothieno[2′,3′:4,5]-
pyrimido[1,2-a]azepine-3-carboxamide (11a)ꢁYield 75%; white
solid; mp 121–123°C; 1H NMR: δ 10.18 (s, 1H), 4.40 (t, Jꢀ=ꢀ4 Hz, 2H), 3.41
(q, J =5 Hz, 2H), 3.07 (t, Jꢀ=ꢀ5 Hz, 2H), 2.85 (s, 3H), 1.84 (dd, Jꢀ=ꢀ20 Hz,
2.6 Hz, 6H), 1.68–1.59 (m, 2H), 1.49–1.39 (m, 2H), 0.96 (t, Jꢀ=ꢀ7 Hz, 3H);
13C NMR: δ 163.4, 161.1, 159.6, 159.2, 145.5, 126.8, 118.5, 43.3, 39.4, 37.1,
31.5, 29.4, 27.4, 25.0, 20.4, 17.3, 13.9. HRMS (ESI). Calcd for C17H23N3O2S
[M–H]−: m/z 333.1516. Found: m/z 333.1511.
N-(2-Chlorophenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
(10l)ꢁYield 87%; light yellow solid; mp 182–184°C; H NMR: δ 12.08
(s, 1H), 8.15 (d, Jꢀ=ꢀ8 Hz, 1H), 7.42 (dd, Jꢀ=ꢀ8, 1.1 Hz, 1H), 7.34–7.27 (m,
1H), 7.09 (td, Jꢀ=ꢀ8 Hz, 1.4 Hz, 1H), 4.11 (t, Jꢀ=ꢀ6 Hz, 2H), 3.03 (t, Jꢀ=ꢀ6 Hz,
2H), 2.88 (s, 3H), 2.08–2.02 (m, 2H), 1.99–1.92 (m, 2H); 13C NMR: δ
161.8, 161.6, 159.5, 155.1, 147.2, 135.4, 127.1, 126.4, 126.0, 125.4, 125.2,
118.3, 43.2, 31.5, 22.1, 19.0, 17.6. HRMS (ESI). Calcd for C18H16ClN3O2S
[M–H]−: m/z 373.0645. Found: m/z 373.0652.
N-(sec-Butyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahydro-
thieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11b)ꢁYield 82%; light yellow solid; mp 153–155°C; 1H NMR: δ 9.96 (s,
1H), 4.41 (t, Jꢀ=ꢀ4 Hz, 2H), 3.06 (t, Jꢀ=ꢀ4 Hz, 2H), 2.84 (s, 3H), 1.87–1.81
(m, 6H), 1.69–1.54 (m, 2H), 1.25 (d, Jꢀ=ꢀ7 Hz, 3H), 0.98 (t, Jꢀ=ꢀ7 Hz, 3H);
13C NMR: δ 162.8, 161.2, 159.6, 159.2, 145.1, 127.2, 118.6, 46.8, 43.3, 37.1,
29.5, 29.5, 27.5, 25.0, 20.1, 17.3, 10.6. HRMS (ESI). Calcd for C17H23N3O2S
[M–H]−: m/z 333.1518. Found: m/z 333.1511.
N-(4-Bromophenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-
4H-pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide
1
(10m)ꢁYield 94%; white solid; mp 234–236°C; H NMR: δ 13.16 (s,
1H), 7.70 (d, Jꢀ=ꢀ9 Hz, 2H), 7.44 (d, Jꢀ=ꢀ9 Hz, 2H), 4.10 (t, Jꢀ=ꢀ6 Hz, 2H),
3.03 (t, Jꢀ=ꢀ6 Hz, 2H), 2.92 (s, 3H), 2.09–2.03 (m, 2H), 2.01–1.93 (m, 2H);
13C NMR: δ 161.8, 161.2, 160.2, 154.9, 148.1, 138.4, 131.6, 126.0, 121.7,
117.9, 116.0, 77.3, 77.0, 76.7, 43.5, 31.5, 22.1, 19.0, 18.0. HRMS (ESI). Calcd
for C18H16BrN3O2S [M–H]−: m/z 417.0145. Found: m/z 4 1 7.0 147.
N,N-Diisopropyl-2-methyl-4-oxo-4,6,7,8,9,10-hexahydro-
thieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11c)ꢁYield 80%; light yellow solid; mp 178–180°C; 1H NMR: δ 4.45 (t,
Jꢀ=ꢀ4 Hz, 2H), 3.79–3.69 (m, 1H), 3.58–3.48 (m, 1H), 3.06 (t, Jꢀ=ꢀ4 Hz, 2H),
2.43 (s, 3H), 1.93–1.70 (m, 6H), 1.68 (d, Jꢀ=ꢀ7 Hz, 3H), 1.58 (d, Jꢀ=ꢀ7Hz,
3H), 1.13 (t, Jꢀ=ꢀ6 Hz, 6H); 13C NMR: δ 164.9, 161.4, 160.2, 156.9, 131.8,
131.2, 119.7, 51.2, 45.9, 42.4, 37.3, 29.6, 27.7, 25.0, 21.2, 20.8, 20.1, 20.0,
13.7. HRMS (ESI). Calcd for C19H27N3O2S [M–H]−: m/z 361.1830. Found:
m/z 361.1824.
N-(2,6-Dichlorophenyl)-2-methyl-4-oxo-6,7,8,9-tetrahydro-4H-
pyrido[1,2-a]thieno[2,3-d]pyrimidine-3-carboxamide (10n)ꢁYield
91%; light yellow solid; mp 165–167°C; 1H NMR: δ 12.26 (s, 1H), 8.14 (d,
Jꢀ=ꢀ9 Hz, 1H), 7.42 (d, Jꢀ=ꢀ2.3 Hz, 1H), 7.29–7.22 (m, 1H), 4.11 (t, Jꢀ=ꢀ6 Hz,
2H), 3.03 (t, Jꢀ=ꢀ6 Hz, 2H), 2.88 (s, 3H), 2.08–2.01 (m, 2H), 2.00–1.92 (m,
2H); 13C NMR: δ 161.8, 161.4, 159.6, 155.2, 147.9, 134.4, 129.7, 129.2, 127.2,
126.9, 125.8, 125.7, 118.2, 43.3, 31.5, 22.1, 19.0, 17.7. HRMS (ESI). Calcd for
C18H15Cl2N3O2S [M–H]−: m/z 407.0260. Found: m/z 407.0262.
2-Methyl-N-octyl-4-oxo-4,6,7,8,9,10-hexahydrothieno[2′,3′:4,5]
pyrimido[1,2-a]azepine-3-carboxamide (11d)ꢁYield 84%; light
yellow solid; mp 102–104°C; 1H NMR: δ 10.18 (s, 1H), 4.40 (t, Jꢀ=ꢀ4 Hz,
Brought to you by | Göteborg University - University of Gothenburg
Authenticated
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ꢁꢀꢀꢀ
L. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-onesꢂ ꢂ7
N-(4-Chlorophenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
2H), 3.42–3.37 (m, 2H), 3.06 (t, Jꢀ=ꢀ4 Hz, 2H), 2.85 (s, 3H), 1.82–1.78 (m,
6H), 1.68–1.59 (m, 2H), 1.44–1.21 (m, 10H), 0.87 (t, Jꢀ=ꢀ6 Hz, 3H); 13C
NMR: δ 163.4, 161.4, 159.6, 159.3, 145.5, 126.8, 118.5, 43.3, 39.7, 37.1, 31.8,
29.5, 29.4, 29.3, 29.2, 27.5, 27.2, 25.0, 22.7, 17.4, 14.1. HRMS (ESI). Calcd
for C21H31N3O2S [M–H]−: m/z 389.2132. Found: m/z 389.2137.
1
(11k)ꢁYield 82%; light yellow solid; mp 253–255°C; H NMR: δ 13.05
(s, 1H), 7.75 (d, Jꢀ=ꢀ8 Hz, 2H), 7.30 (d, Jꢀ=ꢀ8 Hz, 2H), 4.47 (t, Jꢀ=ꢀ5 Hz, 2H),
3.10 (t, Jꢀ=ꢀ5 Hz, 2H), 2.92 (s, 3H), 2.01–1.76 (m, 6H); 13C NMR: δ 161.6,
161.2, 160.0, 159.7, 148.3, 137.9, 128.7, 128.4, 126.3, 121.5, 118.0, 43.6, 37.1,
29.4, 27.4, 25.0, 18.0. HRMS (ESI). Calcd for C19H18ClN3O2S [M–H]−: m/z
387.0810. Found: m/z 387.0808.
N-Cyclohexyl-2-methyl-4-oxo-4,6,7,8,9,10-hexahydro-
thieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11e)ꢁYield 76%; light yellow solid; mp 186–188°C; 1H NMR: δ 10.06
(d, Jꢀ=ꢀ5 Hz, 1H), 4.41 (t, Jꢀ=ꢀ4 Hz, 2H), 3.06 (t, Jꢀ=ꢀ4 Hz, 2H), 2.83 (s,
3H), 2.04–2.04 (m, 2H), 1.86–1.76 (m, 8H), 1.65–1.62 (m, 1H), 1.50–1.16
(m, 6H); 13C NMR: δ 162.5, 161.3, 159.6, 159.2, 145.2, 127.1, 118.5, 48.4,
43.2, 37.2, 32.9, 29.5, 27.5, 25.8, 25.0, 25.0, 17.3. HRMS (ESI). Calcd for
C19H25N3O2S [M–H]−: m/z 359.1663. Found: m/z 359.1667.
N-(2-Chlorophenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11l)ꢁYield 76%; white solid; mp 173–175°C; 1H NMR: δ 13.08 (s, 1H),
8.21 (d, Jꢀ=ꢀ8 Hz, 1H), 7.45 (d, Jꢀ=ꢀ8, 1H), 7.31–7.24 (m, 1H), 7.12 (t, Jꢀ=ꢀ8, Hz,
1H), 4.26 (t, Jꢀ=ꢀ6 Hz, 2H), 3.13 (t, Jꢀ=ꢀ6 Hz, 2H), 2.93 (s, 3H), 2.08–1.89
(m, 6H); 13C NMR: δ 161.8, 161.6, 159.6, 155.3, 147.2, 135.4, 127.2, 126.5,
126.1, 125.5, 125.2, 118.2, 43.7, 37.1, 29.4, 27.4, 24.9, 18.1. HRMS (ESI).
Calcd for C19H18ClN3O2S [M–H]−: m/z 387.0812. Found: m/z 387.0808.
2-Methyl-4-oxo-N-(o-tolyl)-4,6,7,8,9,10-hexahydrothieno[2′,3′:4,5]
pyrimido[1,2-a]azepine-3-carboxamide (11f)ꢁYield 86%; light yel-
low solid; mp 191–193°C; 1H NMR: δ 12.06 (s, 1H), 7.78 (d, Jꢀ=ꢀ8 Hz, 1H),
7.22 (dt, Jꢀ=ꢀ7 Hz, 4 Hz, 2H), 7.08 (t, Jꢀ=ꢀ7 Hz, 1H), 4.48–4.36 (m, 2H), 3.09
(s, 2H), 2.90 (s, 3H), 2.37 (s, 3H), 1.85 (m, 6H); 13C NMR: δ 161.7, 161.3,
159.7, 159.5, 147.5, 136.5, 131.6, 130.4, 126.5, 126.2, 125.1, 124.8, 118.3,
43.4, 37.1, 29.4, 27.4, 25.0, 18.4, 17.8. HRMS (ESI). Calcd for C20H21N3O2S
[M–H]−: m/z 367.1359. Found: m/z 367.1354.
N-(4-Bromophenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11m)ꢁYield 84%; light yellow solid; mp 263–265°C; 1H NMR: δ 13.08
(s, 1H), 7.71 (d, Jꢀ=ꢀ8 Hz, 2H), 7.44 (d, Jꢀ=ꢀ8 Hz, 2H), 4.47 (t, Jꢀ=ꢀ5 Hz, 2H),
3.10(t, Jꢀ=ꢀ5 Hz, 3H), 2.93 (s, 5H), 1.93–1.83 (m, 6H); 13C NMR: δ 161.7,
161.2, 160.0, 159.7, 148.4, 138.4, 131.7, 126.3, 121.8, 118.0, 116.0, 43.6, 37.2,
29.4, 27.4, 25.0, 18.0. HRMS (ESI). Calcd for C19H18BrN3O2S [M–H]−: m/z
431.0308. Found: m/z 431.0303.
2-Methyl-4-oxo-N-(p-tolyl)-4,6,7,8,9,10-hexahydrothieno[2′,3′:4,5]
pyrimido[1,2-a]azepine-3-carboxamide (11g)ꢁYield 90%; light yel-
low solid; mp 223–225°C; 1H NMR: δ 12.71 (s, 1H), 7.67 (d, Jꢀ=ꢀ8 Hz, 2H),
7.15 (d, Jꢀ=ꢀ8 Hz, 2H), 4.56 (t, Jꢀ=ꢀ4 Hz, 2H), 3.09 (d, Jꢀ=ꢀ5 Hz, 2H), 2.93 (s,
3H), 2.33 (s, 3H), 1.96–1.61 (m, 6H); 13C NMR: δ 161.4, 161.1, 159.8, 159.7,
147.5, 136.6, 133.1, 129.3, 126.8, 120.3, 118.2, 43.5, 37.1, 29.4, 27.4, 25.0, 20.9,
17.9. HRMS (ESI). Calcd for C20H21N3O2S [M–H]−: m/z 367.1360. Found:
m/z 367.1354.
N-(2,4-Dichlorophenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11n)ꢁYield 91%; light yellow solid; mp 201–203°C; 1H NMR: δ 12.11 (s,
1H), 8.16 (d, Jꢀ=ꢀ9 Hz, 1H), 7.42 (d, Jꢀ=ꢀ4 Hz, 1H), 7.25 (t, Jꢀ=ꢀ4 Hz, 1H), 4.46
(t, Jꢀ=ꢀ4 Hz, 2H), 3.12–3.05 (t, Jꢀ=ꢀ4 Hz, 2H), 2.87 (s, 3H), 1.91–1.79 (m, 6H);
13C NMR: δ 161.6, 161.1, 159.4, 155.2, 147.7, 134.5, 129.7, 129.2, 127.2, 126.8,
125.8, 125.8, 118.2, 43.5, 37.2, 29.3, 27.4, 25.0, 18.1. HRMS (ESI). Calcd for
C19H17Cl2N3O2S [M–H]−: m/z 421.0423. Found: m/z 421.0419.
N-(4-Methoxyphenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11h)ꢁYield 79%; white solid; mp 200–202°C; 1H NMR: δ 12.67 (s, 1H),
7.70 (d, Jꢀ=ꢀ9 Hz, 2H), 6.89 (d, Jꢀ=ꢀ9 Hz, 2H), 4.56 (t, Jꢀ=ꢀ4 Hz, 2H), 3.81
(s, 3H), 3.09 (t, Jꢀ=ꢀ5 Hz, 2H), 2.92 (s, 3H), 1.94–1.60 (m, 6H); 13C NMR:
δ 161.3, 161.0, 159.8, 159.7, 156.0, 147.4, 132.4, 126.7, 121.9, 118.2, 114.0,
55.5, 43.5, 37.1, 29.4, 27.4, 25.0, 17.9. HRMS (ESI). Calcd for C20H21N3O3S
[M–H]−: m/z 383.1310. Found: m/z 383.1304.
N-Benzyl-2-methyl-4-oxo-4,6,7,8,9,10-hexahydrothieno[2′,3′:4,5]
pyrimido[1,2-a]azepine-3-carboxamide (11o)ꢁYield 73%; light
1
yellow solid; mp 116–118°C; H NMR: δ 10.76 (s, 1H), 7.41 (d, Jꢀ=ꢀ8 Hz,
2H), 7.33 (t, Jꢀ=ꢀ8 Hz, 2H), 7.24 (t, Jꢀ=ꢀ8 Hz, 1H), 4.64 (d, Jꢀ=ꢀ5 Hz, 2H),
4.41–4.34 (t, Jꢀ=ꢀ5 Hz, 2H), 3.06 (t, Jꢀ=ꢀ5 Hz, 2H), 2.86 (s, 3H), 1.96–1.61
(m, 6H); 13C NMR: δ 163.5, 161.4, 159.6, 159.3, 146.2, 139.0, 128.4, 127.7,
126.9, 126.4, 118.5, 43.4, 43.3, 37.1, 29.4, 27.4, 25.0, 17.5. HRMS (ESI). Calcd
for C20H21N3O2S [M–H]−: m/z 367.1350. Found: m/z 367.1354.
N-(2,6-Dimethylphenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11i)ꢁYield 86%; white solid; mp 199–201°C; 1H NMR: δ 11.93 (s, 1H),
7.12–7.04 (m, 3H), 4.45 (t, Jꢀ=ꢀ4 Hz, 2H), 3.12 (t, Jꢀ=ꢀ4 Hz, 2H), 2.90 (s,
3H), 2.31 (s, 6H), 1.91–1.82 (m, 6H); 13C NMR: δ 161.6, 161.4, 159.7, 159.6,
147.6, 135.3, 135.2, 127.9, 126.5, 126.1, 118.5, 43.5, 37.1, 29.4, 27.4, 25.0, 18.7,
17.8. HRMS (ESI). Calcd for C21H23N3O2S [M–H]−: m/z 381.1506. Found:
m/z 381.1511.
2-Methyl-4-oxo-N-(pyridin-2-yl)-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(11p)ꢁYield 79%; light yellow solid; mp 212–214°C; 1H NMR: δ 13.29 (s,
1H), 8.44 (d, Jꢀ=ꢀ4 Hz, 1H), 8.40 (d, Jꢀ=ꢀ8 Hz, 1H), 7.75–7.68 (m, 1H), 7.03
(dd, Jꢀ=ꢀ7 Hz, 5 Hz, 1H), 4.53 (t, Jꢀ=ꢀ4 Hz, 2H), 3.08 (t, Jꢀ=ꢀ4 Hz, 2H), 2.92
(s, 3H), 1.87–1.81 (m, 6H); 13C NMR: δ 161.7, 161.5, 160.0, 159.7, 152.7, 148.3,
148.2, 138.1, 126.3, 119.1, 118.1, 114.9, 43.5, 37.2, 29.5, 27.7, 25.0, 17.9. HRMS
(ESI). Calcd for C18H18N4O2S [M–H]−: m/z 354.1156. Found: m/z 354.1150.
N-(4-Acetylphenyl)-2-methyl-4-oxo-4,6,7,8,9,10-hexahy-
drothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
1
(11j)ꢁYield 88%; light yellow solid; mp 203–205°C; H NMR: δ 13.40
2-Methyl-N-(4-methylpyridin-2-yl)-4-oxo-4,6,7,8,9,10-hexa-
hydrothieno[2′,3′:4,5]pyrimido[1,2-a]azepine-3-carboxamide
(s, 1H), 7.97 (d, Jꢀ=ꢀ8 Hz, 2H), 7.90 (d, Jꢀ=ꢀ8 Hz, 2H), 4.50 (t, Jꢀ=ꢀ4 Hz, 2H),
3.11 (t, Jꢀ=ꢀ4 Hz, 2H), 2.95 (s, 3H), 2.59 (s, 3H), 1.92–1.81 (m, 6H); 13C
NMR: δ 197.2, 161.6, 161.4, 160.0, 159.8, 149.0, 143.9, 132.2, 129.6, 126.1,
119.4, 118.0, 43.6, 37.2, 29.4, 27.4, 26.4, 25.0, 18.1. HRMS (ESI). Calcd for
C21H21N3O3S [M–H]−: m/z 395.1300. Found: m/z 395.1304.
1
(11q)ꢁYield 74%; light yellow solid; mp 237–239°C; H NMR: δ 13.21
(s, 1H), 8.27 (d, Jꢀ=ꢀ9 Hz, 2H), 6.86 (d, Jꢀ=ꢀ4 Hz, 1H), 4.55 (t, Jꢀ=ꢀ4 Hz,
2H), 3.08 (t, Jꢀ=ꢀ5 Hz, 2H), 2.92 (s, 3H), 2.39 (s, 3H), 1.86–1.81 (m, 6H);
13C NMR: δ 161.7, 161.5, 160.0, 159.7, 152.7, 149.5, 148.1, 147.9, 126.4, 120.4,
Brought to you by | Göteborg University - University of Gothenburg
Authenticated
Download Date | 1/27/18 10:05 AM

ꢀꢀꢀꢁ
8ꢀ ꢀL. F. Nie et al.: Amide derivatives of tricyclic thieno[2,3-d]pyrimidin-4(3H)-ones
118.2, 115.3, 43.5, 37.2, 29.5, 27.7, 25.0, 21.5, 18.0. HRMS (ESI). Calcd for [6] Lerner, A. B.; Denton, C. R.; Fitzpatrick, T. B. Clinical and experi-
C19H20N4O2S [M–H]−: m/z 368.1315. Found: m/z 368.1307.
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Cell culture
Murine B16 melanoma cell lines (B16F10) were obtained from the
Chinese Academy of Sciences. The cells were grown in Dulbecco’s
Modified Eagle’s Medium (DMEM) (GIBICO, USA) supplemented with
10% heat-inactivated fetal bovine serum (GIBICO, USA), 100 U/mL
penicillin and 100 mg/mL streptomycin (GIBCO, USA) in a humidi-
fied atmosphere with 5% CO2 at 37°C.
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efficiency and PUVA therapeutic response variation in patients
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Synthesis and bioactivity of novel isoxazole chalcone deriva-
tives on tyrosinase and melanin synthesis in murine B16 cells
for the treatment of vitiligo. Bioorg. Med. Chem. 2016, 24,
5440–5448.
[13] Niu, C.; Pang, G. X.; Li, G.; Dou, J.; Nie, L. F.; Himit, H.; Kabas, M.;
Aisa, H. A. Synthesis and biological evaluation of furocoumarin
derivatives on melanin synthesis in murine B16 cells for the
treatment of vitiligo. Bioorg. Med. Chem. 2016, 24, 5960–5968.
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Melanin content assay
Exponentially growing cells were seeded into 6-well plates at a con-
centration of 5ꢀ×ꢀ105 cells per well. After 24 h of incubation at 37°C, the
culture medium was removed and replaced with fresh medium con-
taining the candidate compounds at different concentrations. The
cells were incubated for another 48 h, washed with ice-cold phos-
phate-buffered saline (PBS), followed by lysis with radioimmuno-
precipitation assay (RIPA) buffer for 40 min on ice, and the lysates
were centrifuged at 10 000 g for 20 min. Supernatants containing
protein were subjected to protein assay, and the pellets with intra-
cellular melanin were solubilized in 200 μL of 1 m NaOH for 2 h at
60°C. The melanin amount was determined spectrophotometrically
at 405 nm using a multi-plate reader. The melanin amount was cal-
culated by normalizing the total melanin values with protein content
(abs melanin/L g protein).
[15] Dewal, M. B.; Wani, A. S.; Vidaillac, C.; Oupicky, D.; Rybak, M. J.;
Firestine, S. M. Thieno[2,3-d]pyrimidinedione derivatives as
antibacterial agents. Eur. J. Med. Chem. 2012, 51, 145–153.
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A. Recent developments regarding the use of thieno[2,3-d]
pyrimidin-4-one derivatives in medicinal chemistry, with a
focus on their synthesis and anticancer properties. Eur. J. Med.
Chem. 2015, 102, 552–573.
Acknowledgments: This work was supported by the
National Natural Science Foundation of China, Grant
No. 21550110495 and the Chinese Academy of Sciences
President’s International Fellowship Initiative (Grant No.
2016PT014).
[17] Narender, M.; Jaswanth, B.; Umashankar, K.; Malathi, J.;
Reddy, A. R.; Umadevi, K. R.; Dusthakeer, A. V. N.; Rao, K. V.;
Raghuram, A. Synthesis, in vitro antimycobacterial evalua-
tion and docking studies of some new 5,6,7,8-tetrahydro-
pyrido[4′,3′:4,5]thieno[2,3-d]pyrimidin-4(3H)-one schiff bases.
Bioorg. Med. Chem. Lett. 2016, 26, 836–840.
[18] Narasimhan, B.; Kumari, M.; Jain, N.; Dhake, A.; Sundaravelan,
C. Correlation of antibacterial activity of some N-[5-(2-furanyl)-
2-methyl-4-oxo-4H-thieno[2,3-d]pyrimidin-3-yl]-carboxamide
and 3-substituted-5-(2-furanyl)-2-methyl-3H-thieno[2,3-d]
pyrimidin-4-ones with topological indices using Hansch analy-
sis. Bioorg. Med. Chem. Lett. 2006, 16, 4951–4958.
[19] Lilienkampf, A.; Karkola, S.; Alho-Richmond, S.; Koskimies, P.;
Johansson, N.; Huhtinen, K.; Vihko, K.; Wähälä, K. Synthesis
and biological evaluation of 17β-hydroxysteroid dehydroge-
nase type 1 (17β-HSD1) inhibitors based on a thieno[2,3-d]
pyrimidin-4(3H)-one core. J. Med. Chem. 2009, 52, 6660–6671.
[20] Kim, H. J.; Kim, J. S.; Woo, J. T.; Lee, I. S.; Cha, B. Y. Hyperpig-
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