Synthesis, characterization and anti-proliferation activities of novel cyano oximino sulfonate esters

Article abstract of DOI:10.1248/cpb.c13-01005

A series of novel cyano oximino sulfonate derivatives were prepared from the reaction of arylsulfonyl chloride with different cyanoacetamide-based oximes ranging from the simplest unsubstituted amide to analogues containing N-ethyl (mimicking the Oxyma template), N-piperidinyl and N-morpholinyl chains. In addition, the cyano oximes, N-hydroxybenzimidoyl cyanide and N-hydroxypicolinimidoyl cyanide were also used in the synthesis of the novel cyano oximino sulfonate derivatives. The structures of the prepared compounds were confirmed by ¹H-NMR, ¹³C-NMR, and elemental analysis. The preliminary bioassays showed that some of the title compounds, such as 2-oxo-2-(piperidin-1-yl)-N-(tosyloxy)acetimidoyl cyanide (TsPipOx), N-(tosyloxy)benzimidoyl cyanide (TsPhOX), N-(naphthalen-2-ylsulfonyloxy)-2-oxo- 2-(piperidin-1-yl)acetimidoyl cyanide (NpsPipOx), 2-amino-N-(naphthalen-2- ylsulfonyloxy)-2-oxoacetimidoyl cyanide (NpsAmOx), N-(naphthalen-2- ylsulfonyloxy)benzimidoyl cyanide (NpsPhCN), and N-(naphthalen-2-ylsulfonyloxy) picolinimidoyl cyanide (NpsPyCN), showed anti-proliferation effect on the mouse fibroblast L929. The calculated IC₅₀-values were ranging between 36.5 μg/mL and 0.235 mg/mL. However the anti-proliferation effects seem to be cytostatic rather than cytotoxic. The compounds only minimize the growth activity without completely killing the cells.

Full text of DOI:10.1248/cpb.c13-01005

April 2014
Regular Article
Chem. Pharm. Bull. 62(4) 373–378 (2014)
373
Synthesis, Characterization and Anti-proliferation Activities of Novel
Cyano Oximino Sulfonate Esters
,c
Ayman El-Faham,a,b Yasser Abbas Elnakdy,* Sarah Abdou Mohamed El Gazzar,b
,b
Mohamed Mokbel Abd El-Rahman,b and Sherine Nabil Khattab*
a Department of Chemistry, College of Science, King Saud University; c Department of Zoology, College of Science,
King Saud University; P.O. Box 2455, Riyadh 11451, Kingdom of Saudi Arabia: and b Department of Chemistry,
Faculty of Science, Alexandria University; P.O. Box 426, Ibrahimia, Alexandria 21321, Egypt.
Received December 28, 2013; accepted February 1, 2014
A series of novel cyano oximino sulfonate derivatives were prepared from the reaction of arylsulfonyl
chloride with different cyanoacetamide-based oximes ranging from the simplest unsubstituted amide to
analogues containing N-ethyl (mimicking the Oxyma template), N-piperidinyl and N-morpholinyl chains. In
addition, the cyano oximes, N-hydroxybenzimidoyl cyanide and N-hydroxypicolinimidoyl cyanide were also
used in the synthesis of the novel cyano oximino sulfonate derivatives. The structures of the prepared com-
1
pounds were confirmed by H-NMR, 13C-NMR, and elemental analysis. The preliminary bioassays showed
that some of the title compounds, such as 2-oxo-2-(piperidin-1-yl)-N-(tosyloxy)acetimidoyl cyanide (TsPipOx),
N-(tosyloxy)benzimidoyl cyanide (TsPhOX), N-(naphthalen-2-ylsulfonyloxy)-2-oxo-2-(piperidin-1-yl)acetimi-
doyl cyanide (NpsPipOx), 2-amino-N-(naphthalen-2-ylsulfonyloxy)-2-oxoacetimidoyl cyanide (NpsAmOx),
N-(naphthalen-2-ylsulfonyloxy)benzimidoyl cyanide (NpsPhCN), and N-(naphthalen-2-ylsulfonyloxy)picolin-
imidoyl cyanide (NpsPyCN), showed anti-proliferation effect on the mouse fibroblast L929. The calculated
IC50-values were ranging between 36.5µg/mL and 0.235mg/mL. However the anti-proliferation effects seem
to be cytostatic rather than cytotoxic. The compounds only minimize the growth activity without completely
killing the cells.
Key words phenylsulfonate; naphthylsulfonate; oxime; anti-proliferation; 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide assay (MTT-assay)
Oximes are one of the most versatile building blocks in the presence of electron-withdrawing substituents in both
organic and organometallic chemistry. The substituent pend- the HOBt 1 and the sulfonic acid moieties.31–36) 1-Hydroxy-
ing at α-position account for the specific properties of each 7-azabenzotriazole (HOAt, 2) has been reported to be more
oxime, such as dissociation constant, solubility, chelating abil- efficient than HOBt 1 because of the anchimeric assistance
ity, etc.1–3) Their multifaceted nature derives in a broad range effect caused by the pyridine ring.37,38)
of applications in chemistry in addition to be biologically
The explosive character of HOBt 1 and its derivatives,39)
prompted the reevaluation of the strongly acidic oxime ethyl
relevant moieties.4–7)
Biologically active heterocyclic compounds containing 2-cyano-2-(hydroxyimino) acetate (Oxyma, 3) as a potential
amine moiety, specifically piperidine and morpholine skel- replacement (Fig. 1).
etons are attractive targets of organic synthesis owing to their
Recently we reported40) the synthesis of sulfonate esters
pharmacological activity and wide occurrence in nature.8,9) based on ethyl cyanoglyoxylate-2-oxime as well as 1-hydroxy-
Piperidines are known to have central nervous system (CNS) 2-pryridinone as new coupling agents. The biological activity
depressant activity at low dosage levels but stimulant activity studies of some selected sulfonate ester derivatives (Fig. 2)
with increased doses. In addition, the piperidine skeleton pos- showed a moderate anti-proliferative effect against human
sesses analgesic, ganglionic blocking and anesthetic proper- cancer cell line SW756 for compounds 4 and 5, while the sul-
ties.10,11) Morpholine derivatives have been reported to possess fonate ester 6 showed a strong growth inhibition for mosquito
antiinflammatory12–14) antimicrobial15,16) and central nervous Culex pipiens 4th instar larvae.41)
system activities.17,18)
We are extending our work by the synthesis of a series of
The sulfonate ester is considered an important class in the sulfonate ester derivatives of the recently prepared N-alkyl-
sulfonylation reaction for the synthesis of sulfonamide which cyanoacetamido oximes 8–11 (Fig. 3), N-hydroxybenzimidoyl
exhibits a wide range of biological activities.19–22)
cyanide 12, and N-hydroxypicolinimidoyl cyanide 13 (Fig. 4).
Sulfonylation is also used as a protection technique for OH
The structure of the target sulfonate ester derivatives were
and NH functionalities, as it enables crystallization and the confirmed by their spectral characterization. The biological
protecting group can be selectively cleaved in the presence
of acid-labile tert-butyl-based groups by using HCl or HBr in
alcohol23) or in aprotic solvents such as ethylacetate or dimeth-
ylformamide.24,25)
Esters of sulfonic acids and 1-hydroxybenzotriazole (HOBt,
1) have been used for peptide coupling.26–30) The reactivity
of these sulfonate esters was shown to be directly related to
The authors declare no conflict of interest.
Fig. 1. Structure of N-Hydroxylamines Used as Additives
© 2014 The Pharmaceutical Society of Japan
*To whom correspondence should be addressed. e-mail: yelnakady@ksu.edu.sa;
Sh.N.Khattab@gmail.com

374
Vol. 62, No. 4
activity study of the target compounds was further investi- oximes 8–11, Chart 1.44–47) Whereas 2-amino-N-hydroxy-
gated. 2-oxoacetimidoyl cyanide (AmOX, 10) and 2-(ethylamino)-N-
We are reporting here the synthesis, spectral characteriza- hydroxy-2-oxoacetimidoyl cyanide (N-Oxyma, 11) precipitated
tion and the biological activity study of a novel series of sulfo- immediately after the acidic treatment, other oximes, such as
nate ester derivatives.
N-hydroxy-2-morpholino-2-oxoacetimidoyl cyanide (MorOX,
The cyanoacetamide oximes 8–11 were prepared using 8) and N-hydroxy-2-oxo-2-(piperidin-1-yl)acetimidoyl cyanide
the Meyer nitrosation reaction.42) Several more convenient (PipOX, 9), needed addition of cold ether, affording white
variations of this reaction are known, using alkyl nitrites crystalline, stable solids in all cases.46,47) N-Hydroxybenz-
such as isobutyl nitrite (iBuONO), isopropyl nitrite (iPrONO) imidoyl cyanide (PhOX, 12) and N-hydroxypicolinimidoyl
and methyl nitrite (MeONO) as source of nitrous acid.1,3,43) cyanide48) (PyOX, 13) were prepared by the nitrosation of
Among these modified procedures, in situ generation of gas- 2-phenylacetonitrile and 2 pyridyl acetonitrile in glacial acetic
eous MeONO was selected in view of the efficacy in previous acid with sodium nitrite.
preparation of oximes.3,44) Thus, nitrosation of 3-morpholino-
The 4-tosyl esters 15–20 and 2-naphthalenesulfonyl esters
3-oxopropanenitrile, 3-oxo-3-(piperidin-1-yl)propanenitrile, 2- 22–27 were prepared via the reaction of 4-tosyl chloride or
cyanoacetamide, and 2-cyano-N-ethylacetamide took place 2-naphthalenesulfonyl chloride with MorOX 8, PipOX 9,
smoothly by this methodology in presence of sodium hydrox- AmOX 10, N-Oxyma 11, PhOX 12 and PyOX 13 in the pres-
ide, followed by acidification to afford the cyanoacetamide ence of triethyl amine (Et3N) in anhydrous dicloromethane
(CH2Cl2) under a nitrogen atmosphere at 0°C40) (Chart 2).
Recrystallization from CH2Cl2-hexane gave pure reagents as
crystals. These reagents are suitable for long-term storage and
are stable toward moisture in the air. IR, elemental analysis
and NMR spectroscopy confirmed the structure of compounds
15–20, 22–27.
The Infrared spectra of compounds 15–20 and 22–27
showed one sharp peak at the range 2234 and 2291cm−1 corre-
sponding to the CN group. In addition, compounds 15–18 and
22–25 showed one sharp peak at the range 1650–1710cm−1
corresponding to the amidic carbonyl group. This is in ac-
cordance with the 13C-NMR spectra of compounds 15–20 and
22–27, which showed a peak at the range 105.96–107.77ppm
Fig. 2. Structure of Compounds 3a,7a-Dihydro-1H-benzo[d][1,2,3]-
triazol-1-yl 4-Methylbenzene Sulfonate (TsOBt, 4), 2-Oxopyridin-
1(2H)-yl 4-Methylbenzenesulfonate (TsOPy, 5), and Ethyl 2-Cyano-
corresponding to the CN group, whereas compounds 15–18
2-(tosyloxyimino)acetate (TsOXY, 6)
and 22–25 showed a peak at the range 154.00–157.24ppm cor-
responding to the amidic carbonyl group.
The anti-proliferation activities of the prepared compounds
have been measured in 3-(4,5-dimethylthiazol-2-yl)-2,5-di-
phenyltetrazolium bromide (MTT)-assay using the mouse
fibroblast L929. The cells were incubated with serial dilutions
of the compounds for 5d (0.003–555µg/mL) in 96-well plates
for 5d then tested for growth inhibitions by use of MTT. The
Fig. 3. Structure of Recently Prepared N-Alkyl-cyanoacetamido Ox-
imes
analysis showed that the tested compounds 16, 19, 23, 24, 26
and 27 showed growth inhibition activity on L929 cells (Fig.
5). The measured IC50-values range between (36.5, 233.5µg/
mL). As shown in Table 1, Compounds 26 and 23 showed the
strongest growth inhibition effects with IC50 values of 36.5
and 51µg/mL, respectively. Compounds 19 and 27 showed a
moderate growth inhibition effects with IC50 values of 93.8
and 107µg/mL respectively. The lowest growth inhibition
activity of the group was measured for compound 16 (IC50
value=233.5 µg/mL). All other tested compounds (15, 17, 18,
Fig. 4. Structure of N-Hydroxybenzimidoyl Cyanide 12, and N-Hy-
droxypicolinimidoyl Cyanide 13
Chart 1. Synthesis of Cyanocarboxamide Oximes 8–11

April 2014
375
20, 22, 25) showed no effect on the growth of L929 cells with IC50 value of 36.5µg/mL. The effect of the naphthyl
under our experimental conditions. moiety was also clear in the oximinopiperidino moiety
In general, the effect of the naphthyl moiety was clear in (NpsPipOx, 23), where its growth inhibition effects with IC50
the oximinopiperidino moiety (NpsPipOx, 23), comparing its values of 51µg/mL compared to its tosyl analogs (TsPipOx,
growth inhibition effects with its tosyl analogs (TsPipOx, 16). 16), which gave the lowest growth inhibition activity (IC50
Further investigations are running in our lab to get insights value=233.5 µg/mL). Further investigations are running in our
into the mode of action of the active compounds.
lab to get insights into the mode of action of the active com-
pounds.
Conclusion
The present work led to development of novel sulfonate
Chemistry
ester derivatives of the recently prepared N-alkyl-cyano-
General 4-Tosyl chloride and 2-naphthalenesulfonyl chlo-
acetamido oximes, N-hydroxybenzimidoyl cyanide, and N- ride were obtained from commercial sources (Aldrich). TLC
hydroxypicolinimidoyl cyanide. The mouse fibroblast L929 was performed on silica gel-protected aluminum sheets (type
was used to measure the cytotoxic activities of the derivatives. 60 GF254, Merck), and the spots were detected by exposure to
The test compound showed different activity against the se- a UV lamp at λ 254nm for a few seconds. Melting points were
lected cell line. However, some of the tested compounds were obtained in open capillary tubes using a Mel-Temp apparatus
inactive. The bioactivities of the active derivatives seem to and were uncorrected. Infrared (IR) spectra were recorded
be rather cytostatic than cytotoxic. The oximinophenyl moi- on a Perkin-Elmer 1600 series Fourier transform instrument
ety in the tosyl series (TsPhOX, 19) gave the highest growth as KBr pellets. Nuclear Magnetic resonance spectra (1H- and
inhibition effects with IC50 value=93.8 µg/mL. The same be- 13C-NMR spectra) were recorded on 400MHz JEOL spec-
havior was observed in the naphthyl derivatives, where the trometer at room temperature. Chemical shifts are reported in
oximinophenyl moiety derivative (NpsPhOx, 26) was the most parts per million (ppm) and are referenced relative to residual
effective one. The combination between the naphthyl and solvent (e.g. CHCl3 at δH 7.26ppm for CDCl3, DMSO at δH
oximinophenyl showed the strongest growth inhibition effects 2.50ppm for DMSO-d6). Spin multiplicities are represented by
the following signals: singlet (s), broad singlet (brs), doublet
(d), broad doublet (brd), doublet of doublets (dd), triplet (t),
doublet of triplets (dt), quartet (q), sextet (sex) and multiplet
(m). Elemental analyses were performed on a Perkin-Elmer
2400 elemental analyzer, and the values found were within
0.3% of the theoretical values. The compounds were il-
lustrated using Chem Draw Ultra version 11, Cambridge Soft
Corporation.
General Procedure for the Preparation of Cyano-
acetamide Oximes (8–11)45) Into an ice-cooled solution
of 2-acetonitrile derivatives, 3-morpholino-3-oxopropaneni-
trile, 3-oxo-3-(piperidin-1-yl)propanenitrile, 2-cyano-N-ethyl-
acetamide or 2-amino-N-hydroxy-2-oxoacetimidoyl cyanide
(100mmol) and sodium hydroxide (4g, 100mmol) in methanol
(30mL) was introduced gaseous methyl nitrite which was gen-
erated from a suspension of sodium nitrite (8.3g) in a mixture
of methanol (10mL) and water (10mL) by dropwise addition
of a mixture of concentrated sulphuric acid (5mL) and water
(10mL). The mixture was stirred for 2h at room temperature
and concentrated to dryness. The residue was dissolved in the
least amount of water (5mL), ice cooled, and acidified with
concentrated hydrochloric acid to precipitate. In some cases
precipitation occurs only by the addition of diethyl ether.
N-Hydroxy-2-morpholino-2-oxoacetimidoyl Cyanide (8):
The product was obtained as white crystals, mp 193–194°C
(lit. 155–160°C)1) in yield 79% (from ethyl cyanoacetic ester).
1H-NMR (C3D6O) δ: 3.66–3.75 (m, 8H, 4CH2), 13.12 (s, 1H,
OH, D2O exchangeable). 13C-NMR (C2D6SO) δ: 43.13, 47.62,
66.22, 66.51, 109.79, 127.31, 158.23.
N-Hydroxy-2-oxo-2-(piperidin-1-yl)acetimidoyl Cyanide (9):
Chart 2. Synthesis of 4-Tosyl Esters 15–20 and 2-Naphthalenesulfonyl
Esters 22–27
Table 1. Substances IC50 Values Measured by MTT-Assay
Compounds
15
16
17
18
19
20
22
23
24
25
26
27
IC50 (µg/mL)
NDa)
233.5
NDa)
NDa)
93.8
NDa)
NDa)
51.0
108.8
NDa)
36.5
107.0
a) Not determined.

376
Vol. 62, No. 4
Fig. 5. Effect of Compounds 16, 19, 23, 24, 26 and 27 on the Proliferation of Mouse Fibroblast L929 as Measured in MTT-Assay
The product was obtained as a white powder, mp 159–160°C of N2 and treated slowly with 5mmol of 4-tosyl chloride 14 or
(lit. 159–162°C)1,49) in yield 87% (from ethyl cyanoacetic 2-naphthalenesulfonyl chloride 21. The reaction mixture was
1
ester). H-NMR (C3D6O) δ: 1.59–1.71 (m, 6H, 3CH2), 3.60– stirred at 0°C for 30min and then at room temperature for 2h.
3.64 (m, 4H, 2CH2). 13C-NMR (C2D6SO) δ: 24.14, 25.59, 26.19, After dilution with 30mL of CH2Cl2, the organic phase was
43.66, 48.03, 109.79, 127.44, 157.73.
washed with water and saturated aqueous NaCl (30mL) and
2-Amino-N-hydroxy-2-oxoacetimidoyl Cyanide (10): The dried over Na2SO4 anhydrous. After removal of the solvent
product was obtained as a white powder, mp 199–200°C in with a rotary evaporator, the residue was recrystallized from
1
yield 84%. H-NMR (CDCl3) δ: 7.07, 7.37 (2 brs, 2H, NH2), CH2Cl2/hexane to give the sulfonate esters.
13.38 (s, 1H, OH, D2O exchangeable). 13C-NMR (C2D6SO) δ:
2-Morpholino-2-oxo-N-(tosyloxy)acetimidoyl
Cyanide
109.45, 128.58, 160.32.
(TsMorOx, 15): This compound was obtained as white crys-
2-(Ethylamino)-N-hydroxy-2-oxoacetimidoyl Cyanide (11): tals, 1.54g (91.3%), mp 118–119°C; IR (KBr) cm−1: 2238
1
The product was obtained as white crystals, mp 174–175°C in (CN), 1650 (CO, amide), 1387, 1193 (SO2, sulfonate). H-NMR
yield 77% (from ethyl cyanoacetic ester). H-NMR (C3D6O) δ: (400MHz, CDCl3) δ: 2.47 (3H, s, CH3), 3.50–3.51 (2H, m,
1
1.14 (t, 3H, CH3, J=7.2Hz), 3.34 (quart., 2H, CH2, J=7.2Hz), CH2), 3.63–3.73 (6H, m, 3CH2), 7.39 (2H, d, Ar-H, J=8.8Hz),
7.83 (brs, 1H, NH), 13.59 (s, 1H, OH, D2O exchangeable). 7.84 (1H, d, Ar-H, J=8.8Hz). 13C-NMR (100MHz, CDCl3) δ:
13C-NMR (C3D6O) δ: 14.21, 34.47, 108.39, 157.96, 158.01.
21.92, 21.99, 43.54, 47.56, 66.38, 66.59, 106.47, 129.37, 130.32,
General Method for the Synthesis of Sulfonate Esters 130.40, 130.44, 147.32, 155.15. Anal. Calcd for C14H15N3O5S: C,
(15–20, 22–27) To a suspension of 5mmol of oxymes 8–13 49.84; H, 4.48; N, 12.46. Found: C, 50.07; H, 4.67; N, 12.28.
in 30mL of anhydrous CH2Cl2 was added 0.71mL (5mmol) of
2-Oxo-2-(piperidin-1-yl)-N-(tosyloxy)acetimidoyl Cyanide
triethylamine with magnetic stirring. The resulting clear yel- (TsPipOx, 16): This compound was obtained as white crystals,
low solution was cooled in an ice bath under an atmosphere 1.49g (87.8%), mp 98–99°C; IR (KBr) cm−1: 2245 (CN), 1654

April 2014
377
1
(CO, amide), 1396, 1189 (SO2, sulfonate). H-NMR (400MHz, acetimidoyl Cyanide (NpsPipOx, 23): This compound was ob-
CDCl3) δ: 1.57–1.69 (6H, m, 3CH2), 2.47 (3H, s, CH3), tained as beige powder, 1.59g (85.7%), mp 75–76°C; IR (KBr)
3.37–3.40 (2H, m, CH2), 3.58–3.61 (2H, m, CH2), 7.39 (2H, cm−1: 2245 (CN), 1655 (CO, amide), 1392, 1183 (SO2, sulfo-
1
d, Ar-H, J=8.8Hz), 7.85 (1H, d, Ar-H, J=8.8Hz). 13C-NMR nate). H-NMR (400MHz, CDCl3) δ: 1.31–1.63 (6H, m, 3CH2),
(100MHz, CDCl3) δ: 21.93, 24.15, 25.33, 26.39, 44.33, 48.38, 3.14–3.30 (2H, m, CH2), 3.54–3.57 (2H, m, CH2), 7.65–7.75
106.60, 129.30, 130.27, 130.67, 132.88, 147.04, 154.76. Anal. (2H, m, Ar-H), 7.79–8.03 (4H, m, Ar-H), 8.58 (1H, s, Ar-H).
Calcd for C15H17N3O4S: C, 53.72; H, 5.11; N, 12.53. Found: C, 13C-NMR (100MHz, CDCl3) δ: 24.08, 25.28, 26.26, 44.32,
53.97; H, 5.39; N, 12.29.
48.32, 106.59, 122.90, 128.22, 128.36, 129.63, 130.07, 130.45,
2-Amino-2-oxo-N-(tosyloxy)acetimidoyl Cyanide (TsAmOx, 131.90, 133.08, 136.02, 154.64. Anal. Calcd for C18H17N3O4S:
17): This compound was obtained as white crystals, 1.19g C, 58.21; H, 4.61; N, 11.31. Found: C, 58.47; H, 4.38; N, 11.09.
(89.1%), mp 105–106°C; IR (KBr) cm−1: 3425, 3243 (NH2),
2-Amino-N-(naphthalen-2-ylsulfonyloxy)-2-oxoacetimidoyl
2236 (CN), 1710 (CO, amide), 1399, 1200 (SO2, sulfonate). Cyanide (NpsAmOx, 24): This compound was obtained as
1H-NMR (400MHz, CDCl3) δ: 2.48 (3H, s, CH3), 6.31, 6.72 white crystals, 1.34g (88.5%), mp 118–120°C; IR (KBr) cm−1:
(2H, brs, 2NH), 7.41 (2H, d, Ar-H, J=8.1Hz), 7.89 (1H, 3439, 3329 (NH2), 2236 (CN), 1696 (CO, amide), 1402, 1192
d, Ar-H, J=8.1Hz). 13C-NMR (100MHz, CDCl3) δ: 22.00, (SO2, sulfonate). 1H-NMR (400MHz, DMSO-d6) δ: 7.65 (1H, t,
105.96, 125.98, 129.07, 129.34, 130.07, 130.58, 133.02, 147.63, Ar-H, J=7.3Hz), 7.84 (1H, t, Ar-H, J=7.3Hz), 8.08–8.30 (6H,
156.92. Anal. Calcd for C10H9N3O4S: C, 44.94; H, 3.39; N, m, 2NH, 4Ar-H), 8.94 (1H, s, Ar-H). 13C-NMR (100MHz,
15.72. Found: C, 45.12; H, 3.55; N, 15.99.
DMSO-d6) δ: 106.68, 128.59, 128.77, 130.08, 130.46, 130.66,
2-(Ethylamino)-2-oxo-N-(tosyloxy)acetimidoyl
Cyanide 131.10, 132.15, 132.59, 136.16, 136.24, 157.24. Anal. Calcd for
(TsN-Oxyma, 18): This compound was obtained as white C13H9N3O4S: C, 51.48; H, 2.99; N, 13.85. Found: C, 51.69; H,
crystals, 1.31g (88.7%), mp 115–116°C; IR (KBr) cm−1: 3.12; N, 13.67.
3359 (NH), 2291 (CN), 1690 (CO, amide), 1396, 1177 (SO2,
2-(Ethylamino)-N-(naphthalen-2-ylsulfonyloxy)-2-oxoacet-
1
sulfonate). H-NMR (400MHz, CDCl3) δ: 1.20 (3H, t, CH3, imidoyl Cyanide (NpsN-Oxyma, 25): This compound was
J=7.3Hz), 2.48 (3H, s, CH3), 3.40 (2H, q, CH2, J=7.3Hz), 6.58 obtained as white crystals, 1.39g (83.7%), mp 114–115°C;
(H, brs, NH), 7.41 (2H, d, Ar-H, J=8.0Hz), 7.87 (1H, d, Ar-H, IR (KBr) cm−1: 3302 (NH), 2235 (CN), 1676 (CO, amide),
J=8.0Hz). 13C-NMR (100MHz, CDCl3) δ: 14.46, 21.97, 35.38, 1400, 1185 (SO2, sulfonate). 1H-NMR (400MHz, CDCl3) δ:
106.08, 129.22, 130.28, 130.50, 133.76, 147.50, 155.01. Anal. 1.17 (3H, t, CH3, J=7.4Hz), 3.40 (2H, q, CH2, J=7.4Hz),
Calcd for C12H13N3O4S: C, 48.81; H, 4.44; N, 14.23. Found: C, 6.62 (H, brs, NH), 7.68 (1H, t, Ar-H, J=7.3Hz), 7.68 (1H, t,
48.62; H, 4.59; N, 13.97.
Ar-H, J=7.3Hz), 7.89–8.05 (4H, m, Ar-H), 8.60 (1H, s, Ar-H).
N-(Tosyloxy)benzimidoyl Cyanide (TsPhCN, 19): This 13C-NMR (100MHz, CDCl3) δ: 14.41, 35.39, 106.11, 122.71,
compound was obtained as whiter crystals, 1.28g (85.3%), 128.26, 128.48, 129.70, 130.06, 130.30, 130.63, 131.88, 131.96,
mp 110–111°C; IR (KBr) cm−1: 2242 (CN), 1393, 1187 (SO2, 133.97, 136.15, 154.94. Anal. Calcd for C15H13N3O4S: C, 54.37;
1
sulfonate). H-NMR (400MHz, CDCl3) δ: 2.46 (3H, s, CH3), H, 3.95; N, 12.68. Found: C, 54.63; H, 3.79; N, 12.87.
7.39 (2H, d, Ar-H, J=8.8Hz), 7.46 (2H, t, Ar-H), 7.57 (1H, t,
N-(Naphthalen-2-ylsulfonyloxy)benzimidoyl
Cyanide
Ar-H, J=7.3Hz), 7.78 (2H, d, Ar-H, J=8.0Hz), 7.94 (1H, d, (NpsPhCN, 26): This compound was obtained as white crys-
Ar-H, J=8.0Hz). 13C-NMR (100MHz, CDCl3) δ: 21.92, 107.77, tals, 1.45g (86.3%), mp 133–134°C; IR (KBr) cm−1: 2234
1
127.32, 127.54. 129.31, 129.44, 130.15, 131.22, 133.47, 139.35, (CN), 1390, 1186 (SO2, sulfonate). H-NMR (400MHz, CDCl3)
146.53. Anal. Calcd for C15H12N2O3S: C, 59.99; H, 4.03; N, δ: 7.44 (2H, t, Ar-H, J=8.1Hz), 7.54 (1H, t, Ar-H, J=7.3Hz),
9.33. Found: C, 60.16; H, 4.25; N, 9.05.
7.65–7.78 (4H, m, Ar-H), 7.93–8.05 (4H, m, Ar-H), 8.67 (1H, s,
N-(Tosyloxy)picolinimidoyl Cyanide (TsPyCN, 20): This Ar-H). 13C-NMR (100MHz, CDCl3) δ: 107.75, 123.23, 127.23,
compound was obtained as grey crystals, 1.35g (89.7%), mp 127.57, 128.14, 129.41, 129.70, 129.83, 130.14, 131.11, 131.68,
128–129°C; IR (KBr) cm−1: 2247 (CN), 1394, 1173 (SO2, 132.01, 133.51, 135.91, 139.60. Anal. Calcd for C18H12N2O3S: C,
1
sulfonate). H-NMR (400MHz, CDCl3) δ: 2.46 (3H, s, CH3), 64.27; H, 3.60; N, 8.33. Found: C, 64.11; H, 3.35; N, 8.05.
7.39–7.47 (3H, m, Ar-H), 7.78–7.95 (4H, m, Ar-H), 8.72 (1H,
N-(Naphthalen-2-ylsulfonyloxy)picolinimidoyl
Cyanide
d, Ar-H, J=4.4Hz). 13C-NMR (100MHz, CDCl3) δ: 21.92, (NpsPyCN, 27): This compound was obtained as grey crys-
107.38, 121.74, 126.90. 129.32, 130.24, 130.97, 137.28, 140.06, tals, 1.50g (89.0%), mp 135–136°C; IR (KBr) cm−1: 2237 (CN),
146.79, 146.86, 150.43. Anal. Calcd for C14H11N3O3S: C, 55.80; 1392, 1185 (SO2, sulfonate). 1H-NMR (400MHz, CDCl3) δ:
H, 3.68; N, 13.95. Found: C, 55.96; H, 3.46; N, 14.09.
7.43–7.45 (1H, m, Ar-H), 7.66–8.05 (8H, m, Ar-H), 8.67–8.72
2-Morpholino-N-(naphthalen-2-ylsulfonyloxy)-2-oxoacet- (2H, m, Ar-H). 13C-NMR (100MHz, CDCl3) δ: 107.34, 121.82,
imidoyl Cyanide (NpsMorOx, 22): This compound was 123.13, 126.91, 128.18, 128.22, 129.70, 129.95, 130.26, 130.87,
obtained as beige powder, 1.60g (85.8%), mp 95–96°C; IR 131.76, 132.00, 135.97, 137.30, 140.23, 146.72, 150.41. Anal.
(KBr) cm−1: 2240 (CN), 1661 (CO, amide), 1376, 1184 (SO2, Calcd for C17H11N3O3S: C, 60.52; H, 3.29; N, 12.46. Found: C,
sulfonate). 1H-NMR (400MHz, CDCl3) δ: 3.38–3.40 (2H, 60.76; H, 3.47; N, 12.19.
m, CH2), 3.48–3.51 (2H, m, CH2), 3.64–3.69 (4H, m, 2CH2),
7.68–7.78 (2H, m, Ar-H), 7.85–7.91 (1H, m, Ar-H), 7.96–8.07
Biology
Cell Culture L929 (mouse fibroblast) cells was obtained
(3H, m, Ar-H), 8.58 (1H, s, Ar-H). 13C-NMR (100MHz,
CDCl3) δ: 43.49, 47.46, 66.33, 66.44, 106.41, 128.24, 129.64, from the American Type Culture Collection, and cultivated
130.28, 130.64, 131.90, 131.95, 136.04, 154.00. Anal. Calcd for under the condition described by their respective depositor.
C17H15N3O5S: C, 54.68; H, 4.05; N, 11.25. Found: C, 54.57; H, Cell culture reagents were purchased from Life Technologies
4.34; N, 11.48.
Inc. (GIBCO BRL). Plastic ware was obtained from Nunc and
N-(Naphthalen-2-ylsulfonyloxy)-2-oxo-2-(piperidin-1-yl)- Saarstedt.

378
Vol. 62, No. 4
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Acknowledgments The work in Egypt was supported
by the Science and Technology Development Fund (STDF)
through the Research Project TC/12/ RSG /2012 (Proposal ID
(4769). The authors thank the Deanship of Scientific Research
at King Saud University for funding this work through re-
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