Synthesis and biological activity of novel diphenyl N - Substituted Carbamimidoylphosphoramidate derivatives

Article abstract of DOI:10.1080/10426507.2012.745075

An efficient synthesis of a series of novel diphenyl N-substituted carbamimidoyl phosphoramidate derivatives was accomplished in two steps. Diphenyl phosphorochloridate (1) was reacted with cyanamide (2) in the presence of 1,4-dimethylpiperazine as a base in THF at 50-55 °C to form an intermediate, diphenyl cyanophosphoramidate (3). Subsequently 3 was reacted with various aromatic/heterocyclic amines (4) in the presence of 1,4-dimethyl piperazine as a base at 55-60 °C to form the corresponding title compounds 5(a-k). The title compounds were tested for antiviral activity against Tobacco mosaic virus (TMV) in vitro, antibacterial and antifungal activities at two different concentrations of 50 and 100 μg/mL. The title compounds exhibited good antiviral and antimicrobial activities when compared to the standards.

Full text of DOI:10.1080/10426507.2012.745075

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Synthesis and Biological Activity
of Novel Diphenyl N- Substituted
Carbamimidoylphosphoramidate
Derivatives
Ch. Subramanyam a , K. Venkata Ramana a , S. Rasheed a , S. Adam b
& C. Naga Raju a
a Department of Chemistry , Sri Venkateswara University , Tirupati ,
Andhra Pradesh , India
b Department of Biochemistry , Sree Vidyanikethan P.G. College ,
Tirupati , Andhra Pradesh , India
Accepted author version posted online: 10 Dec 2012.Published
online: 16 Aug 2013.
To cite this article: Ch. Subramanyam , K. Venkata Ramana , S. Rasheed , S. Adam &
C. Naga Raju (2013) Synthesis and Biological Activity of Novel Diphenyl N- Substituted
Carbamimidoylphosphoramidate Derivatives, Phosphorus, Sulfur, and Silicon and the Related
Elements, 188:9, 1228-1235, DOI: 10.1080/10426507.2012.745075
To link to this article: http://dx.doi.org/10.1080/10426507.2012.745075
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Phosphorus, Sulfur, and Silicon, 188:1228–1235, 2013
Copyright ꢀC Taylor & Francis Group, LLC
ISSN: 1042-6507 print / 1563-5325 online
DOI: 10.1080/10426507.2012.745075
SYNTHESIS AND BIOLOGICAL ACTIVITY
OF NOVEL DIPHENYL N- SUBSTITUTED
CARBAMIMIDOYLPHOSPHORAMIDATE DERIVATIVES
Ch. Subramanyam,1 K. Venkata Ramana,1 S. Rasheed,1
S. Adam,2 and C. Naga Raju1
1Department of Chemistry, Sri Venkateswara University, Tirupati,
Andhra Pradesh, India
2Department of Biochemistry, Sree Vidyanikethan P.G. College, Tirupati,
Andhra Pradesh, India
GRAPHICAL ABSTRACT
Abstract An efficient synthesis of a series of novel diphenyl N-substituted carbamimidoyl
phosphoramidate derivatives was accomplished in two steps. Diphenyl phosphorochloridate
(1) was reacted with cyanamide (2) in the presence of 1,4–dimethylpiperazine as a base in
THF at 50–55 ◦C to form an intermediate, diphenyl cyanophosphoramidate (3). Subsequently
3 was reacted with various aromatic/heterocyclic amines (4) in the presence of 1,4–dimethyl
piperazine as a base at 55–60 ◦C to form the corresponding title compounds 5(a-k). The title
compounds were tested for antiviral activity against Tobacco mosaic virus (TMV) in vitro,
antibacterial and antifungal activities at two different concentrations of 50 and 100 μg/mL.
The title compounds exhibited good antiviral and antimicrobial activities when compared to
the standards.
[Supplementary materials are available for this article. Go to the publisher’s online edition of
Phosphorus, Sulfer, and Silicon and the Related Elements for the following free supplemental
files: Additional text, tables, and figures.]
Keywords Phosphoramidate derivatives; diphenyl phosphorochloridate; cyanamide; aro-
matic/heterocyclic amines; antiviral (TMV); antimicrobial activity
Received 19 August 2012; accepted 27 October 2012.
The authors express their thanks to University of Hyderabad, Hyderabad for providing the spectral data.
Address correspondence to C. Naga Raju, Department of Chemistry, Sri Venkateswara University, Tirupati,
Andhra Pradesh, India. E-mail: rajuchamarthi10@gmail.com
1228

NOVEL BIO ACTIVE PHOSPHORAMIDATE DERIVATIVES
1229
INTRODUCTION
The substituted phosphoramidates, as a class of orgnophosphorus compounds, exhibit
a wide range of biological activities.1,2 An example of a natural phosphoramidate is phos-
phocreatine, which is used as a cardioprotective agent. Phosphoramidates play an important
role in a wide variety of structurally diverse natural and biologically active compounds,
from glycolipids to nucleic acids.3 They have biological activities, such as anti-HIV, an-
tiviral (Reverse-Transcriptase (RT), DNA polymerase, and protease) inhibiting properties.4
Inter nucleotide aminoalkylphosphoramidate moieties are well established as potential an-
tisense or antigene agents.5 Phosphoramidates derived from nucleoside analogues were the
subjects of intensive studies due to their promising prodrug activity against various viruses
(particularly HIV)6 and in cancer therapy.7 The aryloxy substituted phosphoramidates were
found to exhibit enhanced activity against HIV-1 and HIV-2 in cell culture compared to
their parent ddN’s with full retention of activity in thymidinekinase deficient cell lines.8–10
The phosphoramidate of 2-arylquinolone derivatives and related compounds has been the
subject of extensive study as potent cytotoxic antimitotic agents,11 antibacterial agents,12
anti-platelet agents and as cardiovascular protectors.13 It is familiar that phosphorylation
and dephosphorylation of proteins play a very important role in regulating complicated
biochemical processes in all living organisms.14 It is widely used as prodrug moieties
to enhance water solubility15 and therapeutic potential of a parent drug.16–18 In view of
this, we have synthesized and accomplished a new class of novel diphenyl N-substituted
carbamimidoylphosphoramidate derivatives and tested for their efficacy in inhibiting the
growth of bacteria, fungi, and antiviral activity of the plant virus Tobacco mosaic virus
(TMV).
RESULTS AND DISCUSSION
Synthesis
The synthetic route involves two steps. The first step involves the reaction of diphenyl
phosphorochloridate (1) with cyanamide (2) in dry THF in the presence of 1,4-dimethyl
piperazine as a base in THF at 50–55 ◦C to yield diphenylcyanophosphoramidate (3). In the
second step, compound 3 was treated with various amines 4(a–k) in dry THF in the presence
of 1,4-dimethylpiperazine at 55–60 ◦C to obtain the title compounds 5(a–k). The reaction
mixture was filtered and the solvent from the filtrate was removed in a rota-evaporator to
yield the crude products. They were further purified by column chromatography. The title
compounds were obtained with moderate to high yields (68–78%) and are represented in
Scheme 1.
The chemical structures of all the title compounds 5(a–k) were characterized by IR,
1H, 13C, 31P NMR, mass spectral data and elemental analyses and their data are presented
in the experimental section. IR absorptions were observed in the regions 3462–3438,
3352–3320, 1660–1628, and 1225–1210 cm−1 are assigned to N–H, C NH, C N, and
P O stretching vibrations respectively for 5(a–k). The 1H NMR spectra gave broad signals
at δ 7.96–7.87 ppm, due to the C NH protons. The proton signals of Ar–H appeared as
multiplet in the range of δ 7.24–6.82. The signals due to P–NH proton appeared as a singlet
in the region δ 5.58–5.40 ppm.13C NMR chemical shifts were observed in the regions, δ
168.8–163.2 for C NH. 31P NMR signals were observed in the region δ 24.9–21.3 ppm19
for all the compounds 5(a–k). All the phosphoramidate derivatives 5(a–k) were screened in
vitro for their antiviral activity against the TMV, antibacterial, and antifungal activities and

1230
CH. SUBRAMANYAM ET AL.
Scheme 1
compared their activities with standard compounds. The title compounds 5a, 5b, 5e, 5k,
and 5i exhibited good efficacy of antiviral and antibacterial activity due to the phosphorus
atom attached to the substituent having –F, –Cl, F, –OCH3, –CF3, and –HNC O groups.
The compounds 5f, 5c, and 5d exhibited good antifungal activity due to the presence

NOVEL BIO ACTIVE PHOSPHORAMIDATE DERIVATIVES
1231
of sulfathiazole, 6-nitrobenzothiazole and 6-methoxybenzothiazole moieties. Additional
results are presented in the Supplemental Materials (Tables S1–S3).
Antiviral Activity
The newly synthesized diphenyl N-substituted carbamimidoylphosphoramidate
derivatives 5(a–k) were screened for their antiviral activity against TMV by the Goodings
method.20,21
Antibacterial Activity
The newly synthesized carbamimidoylphosphoramidate derivatives 5(a–k) were
screened for their antibacterial activity.22,23 The title compounds inhibited the bactria against
the Gram positive bacteria Bacillus subtilis and Enterococcus faecalis and Gram negative
bacteria Pseudomonas aeruginosa and Klebsiella pneumoniae and ampicillin was used as
a reference standard.
Antifungal Activity
The newly synthesized carbamimidoylphosphoramidate derivatives 5(a–k) were
screened for their antifungal activity.22,23 The title compounds 5(a–k) inhibited the fungi
against the Candida albicans and Aspergillus niger.
EXPERIMNENTAL
Chemistry
Chemicals were purchased from Sigma–Aldrich, Merck and Lancaster, and were used
as such without further purification. All solvents used for spectroscopic and other physical
studies were reagent grade and were further purified by literature methods.24 Melting points
were determined using a calibrated thermometer by Guna Digital Melting Point apparatus.
IR Spectra were recorded as KBr discs on a Nicolet 380 FT-IR spectrophotometer. 1H and
13C NMR spectra were recorded as solutions in DMSO-d6 on a Bruker AMX 400 MHz
spectrometer operating at 400 MHz for 1H, 100 MHz for 13C and 161.9 MHz for 31P. The
1H and 13C chemical shifts were referenced to tetramethylsilane and 31P chemical shifts to
85% H3PO4. LC mass spectra were recorded on a Jeol SX 102 DA/600 Mass spectrometer.
Elemental analyses were performed on a Thermo Finnigan Instrument at the University of
Hyderabad, Hyderabad, India.
General Procedure for the Synthesis of Title Compounds 5(a–k)
The synthesis of diphenyl N-substituted carbamimidoylphosphoramidate was
achieved in two steps. In the first step, diphenylphosphorochloridate (1) was reacted with
cyanamide (2) in the presence of 1,4-dimethylpiperazine (DMP) as a base in THF at
◦
50–55 C to form the intermediate diphenylcyanophosphoramidate (3). Formation of 3
was ascertained by TLC analysis run in 3:7 mixture of ethyl acetate and hexane. In the
second step, 3 was treated with aromatic/heterocyclic amines 4(a–k) in the presence of
◦
1,4-dimethylpiperazine at 55–60 C to obtain the title products, diphenyl N-substituted

1232
CH. SUBRAMANYAM ET AL.
carbamimidoylphosphoramidates 5(a–k). Identification of the products and completion of
the reaction was monitored by TLC using ethyl acetate: hexane (3:2). After completion
of the reaction, the hydrochloride salt was filtered off and the solvent was removed in
a rota-evaporator and the residue was purified by column chromatography on silica gel
(100–200 mesh) using petroleum ether–ethyl acetate (3:7) as eluent. The resulting title
compounds 5(a–k) were obtained in high yields (68–78%). The structures of the title
compounds 5(a–k) were established by spectral and elemental analysis. Representative
spectra (1H, 13C, and 31P) for compounds 5a and 5f are presented in the Supplemental
Materials (Figures S1–S6).
Diphenyl N-(4-fluorobenzyl)carbamimidoylphosphoramidate (5a).
Yield: 68%, Semi Solid. IR (KBr) (νmax cm−1): 3458 (–NH), 3352 (C NH), 1642 (C N),
1212 (P O) cm−1; 1H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H, –C NH), 7.42 (d, J =
8.2 Hz, 2H, Arꢀ–H), 7.31 (d, J = 7.4 Hz, 2H, Arꢀ-H), 7.24–6.85 (m, 10H, Ar–H), 5.40 (s,
1H, P-NH), 4.13 (d, J = 5.4 Hz, 2H, –CH2), 2.72 (s, 1H, –CH2–NH); 13C-NMR (100 MHz,
DMSO-d6): δ 151.4 (C-1 & C-7), 120.5 (C-2 & C-8), 129.4 (C-3 & C-9), 122.4 (C-4 &
C-10), 132.4 (C-5 & C-11), 120.6 (C-6 & C-12), 168.2 (C-13), 43.8 (C-14), 130.4 (C-1ꢀ),
127.4 (C-2ꢀ), 114.5 (C-3ꢀ), 162.2 (C-4ꢀ), 114.5 (C-5ꢀ), 127.5 (C-6ꢀ); 31P NMR (DMSO-d6):
δ 22.1; LCMS (m/z,%): 400 (M+H+, 100); Anal. Calcd for C20H19FN3O3P: C, 60.15; H,
4.80; N, 10.52; found C, 60.19; H, 4.76; N, 10.58.
Diphenyl N-(3-chloro-4-fluorophenyl)carbamimidoylphosphorami
date (5b). Yield: 73%, Semi Solid. IR (KBr) (νmax cm−1): 3462 (–NH), 3350 (C NH),
1628 (C N), 1215 (P O); 1H NMR (400 MHz, DMSO-d6): δ 9.90 (s, 1H, Ar–NH), 7.95
(s, 1H, –C NH), 7.45 (d, J = 8.2 Hz, 1H, Arꢀ-H), 7.32 (d, J = 6.4 Hz, 1H, Arꢀ–H), 7.28
(s, 1H, Arꢀ–H), 7.22–6.82 (m, 10H, Ar–H), 5.47 (s, 1H, (P–NH); 13C-NMR (100 MHz,
DMSO-d6): δ 150.2 (C-1 & C-7), 121.6 (C-2 & C-8), 128.2 (C-3 & C-9), 123.5 (C-4 &
C-10), 131.4 (C-5 & C-11), 121.2 (C-6 & C-12), 163.2 (C-13), 134.5 (C-1ꢀ), 124.5 (C-2ꢀ),
118.2 (C-3ꢀ), 152.4 (C-4ꢀ), 118.4 (C-5ꢀ), 126.4 (C-6ꢀ); 31P NMR (DMSO-d6): δ 23.2; LCMS
(m/z,%): 420 (M+H+, 100), 422 (M+2, 33); Anal. Calcd for C19H16ClFN3O3P: C, 54.36;
H, 3.84; N, 10.01; found C, 54.41; H, 3.80; N, 9.94.
Diphenyl N-(6-nitrobenzo[d]thiazol-2-yl)carbamimidoylphosphorami
date (5c). Yield: 70%, Semi Solid. IR (KBr) (νmax cm−1): 3456 (–NH), 3348 (C NH),
1634 (C N), 1217 (P O); 1H NMR (400 MHz, DMSO-d6): δ 9.45 (s, 1H, Ar–NH), 8.32
(d, J = 8.2 Hz, 1H, Arꢀ-H), 8.28 (d, J = 6.8 Hz, 1H, Arꢀ–H), 8.12 (s, 1H, Arꢀ–H), 7.93
(s, 1H, –C NH), 7.20–6.86 (m, 10H, Ar–H), 5.58 (s, 1H, (P–NH); 13C-NMR (100 MHz,
DMSO-d6): δ 151.4 (C-1 & C-7), 121.2 (C-2 & C-8), 127.4 (C-3 & C-9), 123.2 (C-4 &
C-10), 130.2 (C-5 & C-11), 120.8 (C-6 & C-12), 168.8 (C-13), 172.2 (C-1ꢀ), 154.5 (C-2ꢀ)
114.6 (C-3ꢀ), 124.5 (C-4ꢀ), 145.4 (C-5ꢀ), 119.4 (C-6ꢀ), 131.4 (C-7ꢀ); 31P NMR (DMSO-d6):
δ 24.9; LCMS (m/z,%): 470 (M+H+, 100); Anal. Calcd for C20H16N5O5PS: C, 51.17; H,
3.44; N, 14.92; found C, 51.24; H, 3.36; N, 14.85.
Diphenyl N-(6-methoxybenzo[d]thiazol-2yl)carbamimidoylphosphora
midate(5d). Yield: 74%, Semi Solid. IR (KBr) (νmax cm−1): 3452 (–NH), 3346 (C NH),
1647 (C N), 1217 (P O); 1H NMR (400 MHz, DMSO-d6): δ 8.90 (s, 1H, Ar–NH), 8.18
(d, J = 8.4 Hz, 1H, Arꢀ–H), 8.12 (s, 1H, Arꢀ–H), 8.06 (d, J = 7.2 Hz, 1H, Arꢀ–H), 7.88
(s, 1H, –C NH), 7.18–6.94 (m, 10H, Ar–H), 5.56 (s, 1H, (P–NH), 3.86 (s, 3H, OCH3);
13C-NMR (100MHz, DMSO-d6): δ 150.2 (C-1 & C-7), 120.4 (C-2 & C-8), 126.5 (C-3
& C-9), 124.6 (C-4 & C-10), 131.4 (C-5 & C-11), 120.4 (C-6 & C-12), 164.4 (C-13),
172.6 (C-1ꢀ), 153.5 (C-2ꢀ) 116.8 (C-3ꢀ), 123.6 (C-4ꢀ), 146.2 (C-5ꢀ), 118.8 (C-6ꢀ), 130.6
(C-7ꢀ), 56.2 (O-CH3); 31P NMR (DMSO-d6): δ 21.3; LCMS (m/z,%): 455 (M+H+, 100);

NOVEL BIO ACTIVE PHOSPHORAMIDATE DERIVATIVES
1233
Anal. Calcd for C21H19N4O4PS: C, 55.50; H, 4.21; N, 12.33; found C, 55.56; H, 4.26; N,
12.28.
Diphenyl N-(4,6-dimethoxypyrimidin-2-yl)carbamimidoylphosphorami
date (5e). Yield: 69%, Semi Solid. IR (KBr) (νmax cm−1): 3455 (–NH), 3348 (C NH),
1646 (C N), 1210 (P O); 1H NMR (400 MHz, DMSO-d6): δ 8.97 (s, 1H, Ar–NH), 8.14
(s, 1H, Arꢀ–H), 7.87 (s, 1H, –C NH), 7.18–6.92 (m, 10H, Ar–H), 5.43 (s, 1H, (P–NH),
3.80 (s, 6H, –OCH3); 13C-NMR (100 MHz, DMSO-d6): δ 151.4 (C-1 & C-7), 122.2 (C-2
& C-8), 127.4 (C-3 & C-9), 125.2 (C-4 & C-10), 130.5 (C-5 & C-11), 121.5 (C-6 & C-12),
165.2 (C-13), 151.8 (C-1ꢀ), 171.5 (C-2ꢀ & C-4ꢀ), 121.4, (C-3ꢀ), 51.6 (O-CH3); 31P NMR
(DMSO-d6): δ 21.4; LCMS (m/z,%): 430 (M+H+, 100); Anal. Calcd for C19H20N5O5P:
C, 53.15; H, 4.70; N, 16.31; found C, 53.19; H, 4.66; N, 16.28.
Diphenyl N-(4-(N-thiazol-2 ylsulfamoyl) phenyl)carbamimidoylphospho
ramidate (5f). Yield: 70%, Semi Solid. IR (KBr) (νmax cm−1): 3438 (–NH), 3320
1
(C NH), 1636 (C N), 1219 (P O); H NMR (400 MHz, DMSO-d6): δ 12.30 (s, 1H,
SO2–NH), 9.96 (s, 1H, Ar–NH), 8.16 (d, J = 6.4 Hz, 1H, Arꢀ–H), 8.12 (d, J = 8.2 Hz, 1H,
Arꢀ–H), 8.02 (d, J = 7.2 Hz, 2H, Arꢀ–H), 7.96 (s, 1H,–C NH), 7.82 (d, J = 8.4 Hz, 2H,
Arꢀ–H), 7.16–6.84 (m, 10H, Ar–H), 5.45 (s, 1H, (P–NH); 13C-NMR (100 MHz, DMSO-d6):
δ 151.4 (C-1 & C-7), 121.5 (C-2 & C-8), 124.2 (C-3 & C-9), 125.7 (C-4 & C-10), 130.2
(C-5 & C-11), 121.5 (C-6 & C-12), 167.6 (C-13), 147.6 (C-1ꢀ), 118.5 (C-2ꢀ) 130.8, (C-3ꢀ),
128.2 (C-4ꢀ), 116.2 (C-5ꢀ), 118.8 (C-6ꢀ), 169.2 (C-7ꢀ), 131.2 (C-8ꢀ), 116.1 (C-9ꢀ); 31P NMR
(DMSO-d6): δ 24.7; LCMS (m/z,%): 530 (M+H+, 100); Anal. Calcd for C22H20N5O5PS2:
C, 49.90; H, 3.81; N, 13.23; found C, 49.96; H, 3.86; N, 13.18.
Diphenyl
N-(5-chloro-2-phenoxyphenyl)carbamimidoylphosphorami
date (5g). Yield: 78%. Semi Solid. IR (KBr) (νmax cm−1): 3442 (–NH), 3347 (C NH),
1660 (C N), 1218 (P O); 1H NMR (400 MHz, DMSO-d6): δ 9.75 (s, 1H, Ar-NH), 7.92
(s, 1H, –C NH), 7.62 (d, J = 6.6 Hz, 1H, Arꢀ–H), 7.46 (d, J = 9.2 Hz, 1H, Arꢀ–H), 7.38
(s, 1H, Arꢀ–H), 7.21–6.94 (m, 15H, Ar–H), 5.48 (s, 1H, (P-NH); 13C-NMR (100 MHz,
DMSO-d6): δ 148.2 (C-1 & C-7), 120.2 (C-2 & C-8), 125.4 (C-3 & C-9), 126.2 (C-4 &
C-10), 131.4 (C-5 & C-11), 120.4 (C-6 & C-12), 166.2 (C-13), 148.2 (C-1ꢀ), 120.5 (C-2ꢀ)
131.2, (C-3ꢀ), 127.6 (C-4ꢀ), 117.4 (C-5ꢀ), 119.2 (C-6ꢀ), 152.2 (C-7ꢀ), 128.2 (C-8ꢀ), 117.5
(C-9ꢀ), 126.2 (C-10ꢀ), 129.2 (C-11ꢀ), 115.6 (C-12ꢀ); 31P NMR (DMSO-d6): δ 23.1; LCMS
(m/z,%): 494 (M+H+. 100), 496 (M+2, 32); Anal. Calcd for C25H21ClN3O4P: C, 60.80;
H, 4.29; N, 8.51; found C, 60.90; H, 4.35; N, 8.41.
Diphenyl
N-naphthalen-1-ylcarbamimidoylphosphoramidate
(5h).
Yield: 75%, Semi Solid. IR (KBr) (νmax cm−1): 3449 (–NH), 3345 (C NH), 1654 (C N),
1
1215 (P O); H NMR (400 MHz, DMSO-d6): δ 9.85 (s, 1H, Ar–NH), 7.96 (s, 1H,
–C NH), 7.64 (d, J = 8.4 Hz, 1H, Arꢀ–H), 7.42 (d, J = 7.8 Hz, 2H, Arꢀ–H), 7.36 (t, J =
7.2 Hz, 1H, Arꢀ–H), 7.30 (d, J = 9.2 Hz, 1H, Arꢀ–H), 7.26 (d, J = 6.4 Hz, 2H, Arꢀ–H),
7.18–6.86 (m, 10H, Ar–H), 5.41(s, 1H, (P–NH); 13C-NMR (100 MHz, DMSO-d6) δ 149.4
(C-1 & C-7), 120.5 (C-2 & C-8), 127.6 (C-3 & C-9), 128.4 (C-4 & C-10), 130.2 (C-5
& C-11), 122.4 (C-6 & C-12), 164.2 (C-13), 140.4 (C-1ꢀ), 127.2 (C-2ꢀ) 125.6, (C-3ꢀ),
127.2 (C-4ꢀ), 119.4 (C-5ꢀ), 121.2 (C-6ꢀ), 128.2 (C-7ꢀ), 126.4 (C-8ꢀ), 124.5 (C-9ꢀ), 134.2
(C-10ꢀ); 31P NMR (DMSO-d6): δ 23.4; LCMS (m/z,%): 418 (M+H+, 100); Anal. Calcd
for C23H20N3O3P: C, 66.18; H, 4.83; N, 10.07; found C, 66.25; H, 4.88; N, 10.01.
Diphenyl
N-(2-nicotinoylhydrazinyl)carbamimidoylphosphoramidate
(5i). Yield: 74%, Semi Solid. IR (KBr) (νmax cm−1): 3438 (–NH), 3334 (C NH), 1642
(C N), 1219 (P O); 1H NMR (400 MHz, DMSO-d6): δ 10.95 (s, 1H, CO-NH), 7.94 (s,
1H, –C NH), 7.38 (s, J = 7.2 Hz, 1H, Arꢀ–H), 7.28 (d, J = 8.2 Hz, 1H, Arꢀ–H), 7.30

1234
CH. SUBRAMANYAM ET AL.
(d, J = 6.4 Hz, 1H, Arꢀ–H), 7.26 (t, J = 8.4 Hz, 1H, Arꢀ–H), 7.20–6.85 (m, 10H, Ar–H),
6.1 (s, 1H, (C O)–NH–NH), 5.54 (s, 1H, (P–NH); 13C-NMR (100 MHz, DMSO-d6): δ
148.2 (C-1 & C-7), 121.5 (C-2 & C-8), 126.4 (C-3 & C-9), 127.5 (C-4 & C-10), 131.4
(C-5 & C-11), 125.2 (C-6 & C-12), 166.5 (C-13), 164.5 (C-14), 132.5 (C-1ꢀ), 128.7 (C-2ꢀ)
129.5, (C-3ꢀ), 141.4 (C-4ꢀ), 143.6 (C-5ꢀ); 31P NMR (DMSO-d6): δ 24.7; LCMS (m/z,%):
412 (M+H+, 100); Anal. Calcd for C19H18N5O4P: C, 55.48; H, 4.41; N, 17.03; found C,
55.54; H, 4.45; N, 17.01.
Diphenyl N-(4-(4-fluorophenyl)piperazin-1yl)carbamimidoylphosphora
midate (5j). Yield: 72%, Semi Solid. IR (KBr) (νmax cm−1): 3434 (–NH), 3326 (C NH),
1632 (C N), 1220 (P O); 1H NMR (400 MHz, DMSO-d6): δ 7.92 (s, 1H, –C NH), 7.44
(d, J = 7.6 Hz, 2H, Arꢀ–H), 7.34 (d, J = 8.4 Hz, 2H, Arꢀ–H), 7.22–6.86 (m, 10H, Ar–H),
5.92 (s, 1H, –N–NH), 5.51 (s, 1H, P–NH), 3.66 (t, J = 6.4 Hz, 4H, –CH2), 2.88 (t, J =
5.6 Hz, 4H, –CH2); 13C-NMR (100 MHz, DMSO-d6): δ 151.6 (C-1 & C-7), 121.4 (C-2 &
C-8), 124.2 (C-3 & C-9), 129.5 (C-4 & C-10), 130.6 (C-5 & C-11), 121.8 (C-6 & C-12),
163.6 (C-13), 58.2 (C-1ꢀ & C-4ꢀ), 47.6 (C-2ꢀ & C-3ꢀ), 145.2 (C-5ꢀ), 116.4 (C-6ꢀ & C-10ꢀ),
114.6 (C-7ꢀ & C-10ꢀ), 147.2 (C-8ꢀ); 31P NMR (DMSO-d6): δ 23.5; LCMS (m/z,%): 470
(M+H+, 100); Anal. Calcd for C23H25FN5O3P: C, 58.84; H, 5.37; N, 14.92; found C,
58.88; H, 5.41; N, 14.88.
Diphenyl N-(2-chloro-5-(trifluoromethyl)phenyl)carbamimidoylphospho
ramidate (5k). Yield: 76%, Semi Solid. IR (KBr) (νmax cm−1): 3456 (–NH), 3345
1
(C NH), 1652 (C N), 1225 (P O); H NMR (400 MHz, DMSO-d6): δ 9.50 (s, 1H,
Ar–NH), 7.98 (s, 1H, C NH), 7.72 (d, J = 8.6 Hz, 1H, Arꢀ–H), 7.56 (d, J = 9.2 Hz, 1H,
Arꢀ–H), 7.32 (s, 1H, Arꢀ–H), 7.16–6.90 (m, 10H, Ar–H), 5.52 (s, 1H, (P–NH); 13C-NMR
(100 MHz, DMSO-d6): δ 150.2 (C-1 & C-7), 124.5 (C-2 & C-8), 125.4 (C-3 & C-9),
128.2 (C-4 & C-10), 131.5 (C-5 & C-11), 120.4 (C-6 & C-12), 165.8 (C-13), 142.4 (C-1ꢀ),
127.6 (C-2ꢀ) 125.2, (C-3ꢀ), 118.4 (C-4ꢀ), 125.6 (C-5ꢀ), 114.2 (C-6ꢀ), 121.4 (-CF3);31P NMR
(DMSO-d6): δ 24.5; LCMS (m/z,%): 470 (M+H+, 100), 472 (M+2, 32); Anal. Calcd for
C20H16ClF3N3O3P: C, 51.13; H, 3.43; N, 8.94; found C, 51.17; H, 3.47; N, 8.90.
CONCLUSIONS
The synthesis of novel diphenyl N-substituted carbamimidoylphosphoramidates
5(a–k) was accomplished and their structures were confirmed by elemental analysis and
spectral data (IR, 1H, 13C, and 31P NMR, LC- mass). The title compounds 5a, 5b, 5e, 5k,
and 5i exhibited good antiviral, antibacterial and 5f, 5c, 5d showed high antifungal activities
compared to reference standards like nignanmycin, ampicillin, and nystatine respectively.
REFERENCES
1. Manfredini, S.; Baraldi, P. G.; Durini, E.; Vertuani, S.; Balzarini, J.; De Clercq, E.; Karlsson, A.;
Buzzoni, V.; Thelander, L. J. Med. Chem. 1999, 42, 3243-3250.
2. Yang, K. W.; Brandt, J. J.; Chatwood, L. L.; Crowder, M. W. Bioorg. Med. Chem. Lett. 2000, 10,
1085-1087.
3. Jones, S.; Selitsianos, D.; Thompson, K. J.; Toms, S. M. J.Org. Chem. 2003, 68, 5211-5216.
4. Petersen, D.; Marcolini, M.; Bernadi, L.; Fini, F.; Herrera, P. R.; Sgarzani, V.; Ricci, A. J. Org.
Chem. 2006, 71, 6269-6272.
5. Letsinger, R. L.; Singman, C. N.; Histand, G.; Salunkhe, M. J. Am. Chem. Soc. 1988, 110,
4470-4471.

NOVEL BIO ACTIVE PHOSPHORAMIDATE DERIVATIVES
1235
6. Hecker, S. J.; Erion, M. D. J. Med. Chem. 2008, 51, 2328-2345.
7. Iyer, V. V.; Greisgraber, G. W.; Radmer, M. R.; McIntee, E. J.; Wagner, C. R. J. Med. Chem.
2000, 43, 2266-2274.
8. Guigan, C. Mc.; Cahard, D.; Shecka, H. M.; Declercas, E.; Balzarini, J. Bio. Org. Med. Chem
Lett. 1996, 6, 1183-1186.
9. Guigan, C. Mc.; Cahard, D.; Shecka, H. M.; Declercas, E.; Balzarini, J. J. Med. Chem. Lett.
1996, 39, 1948-1952.
10. Guigan, C. Mc.; Tsang, H. W.; Cahard, D.; Turner, S.; Velazquez, S.; Salgado, A.; Bidois, L.;
Naesens, L.; Declercas, E.; Velazquvez, J. Antiviral Res. 1997, 35, 195-201.
11. Rosen, M.; Krebs, E. G. Protein Phosphorylation, Cold Laboratory; Cold Spring Harbor: New
York, 1981, 849-850.
12. Li, L.; Wang, H. K.; Kuo, S. C.; Wu, T. S.; Mauger, A.; Lin, C. M.; Hamel, E.; Kuo, K. H. J.
Med. Chem. 1994, 37, 3400-3407.
13. Huang, L. J.; Hsieh, M. C.; Teng, C. M.; Lee, K. H.; Kuo, S. C. Bioorg. Med. Chem. 1998, 6,
1657-1652.
14. Lee, H. Z.; Lin, W. C.; Yeh, F. T.; Wu, C. H. J. Pharmacol. 1998, 354, 205-213.
15. Fleisher, D.; Bong, R.; Stewart, B. H. Adv. Drug Delivery Res. 1996, 19, 115-130.
16. Chang, S.; Griesgraber, G. W.; Southern, P. J.; Wagner, C. R. J. Med. Chem. 2001, 44, 223-231.
17. Egron, D.; Imbach, J. L.; Gosselin, G.; Aubertin, A. M.; Perigaud, C. J. Med. Chem. 2003, 46,
4564-4571.
18. Freel Meyers, C. L.; Borch, R. F. J. Med. Chem. 2000, 43, 4319-4327.
19. Quin, L. D., Verkade, J. G. Phosphorus-31 NMR Spectral Properties Compound Characterization
and Structural Analysis; VCH Publishers: New York, 1994.
20. Gooding, G. V.; Hebert, T. T. Phytopathol. 1967, 57, 1285-1287.
21. Song, B. A.; Zhang, H. P.; Wang, H.; Yang, S.; Jin, L. H.; Hu, D. Y.; Pang, L. L.; Xue, W. J.
Agric. Food Chem. 2005, 53, 7886-7891.
22. Clinical and Laboratory Standards Institute: Performance Standards for Antimicrobial Suscepti-
bility Testing; Sixteen International Supplements. CLSI document M100 S16. Vol. 26-3; M7-A7,
Vol. 26-2; M2-A9, vol. 26-1. Wayne, PA, USA, 2006.
23. Akarboom, T. P. M.; Sies, H. Assay of glutathione, glutathione disulfide and glutathione mixed
disulfides in biological samples. In: W. B. Jakoby (Ed.), Methods in Enzymology; vol. 77.
Academic Press: New York, 1981, pp. 373-382.
24. Armarego, W. L. F.; Perrin, D. D. Purification of Laboratory Chemicals, 4th ed.; Butterworth:
Heinemann, Oxford, OX2 8DP 1997.