Synthesis and Anti cancer Activity Test against HeLa Cells from N-(2-Chlorobenzoyl)-N’-Phenylurea

  • Bambang Tri Purwanto Surabaya


Research in the search for cancer drug compounds continue to be developed given the lack of specific anticancer compounds. Some urea derived compounds are also continuously being developed in search of potent anticancer compounds with minimal side effects. In relation to the above, we want to develop an urea derivative of N-phenylurea compound which will be reacted with benzoyl chloride derivative that is 2-chlorobenzoyl chloride so that N-(2-chlorobenzoyl) -N'phenylurea will be obtained. Synthesis of N- (2-chlorobenzoyl) -N'phenylurea was done by modified Schotten-Baumman method, then purity test was performed with thin layer chromatography using 3 different  solvents. The next step is structural characterization using UV and IR spectrophotometry method, then 1H-NMR and MS spectrometry, so that the structure of N- (2-chlorobenzoyl) -N'phenylurea  will be obtained. An anticancer activity test is performed on HeLa cells using MTT assay method and IC50 value will be obtained.

Compounds that have been successfully synthesized are compound of N- (2-chlorobenzoyl) -N'phenylurea, with yield of 80.47% in the form of white needle crystal. The purity test of the N- (2-chlorobenzoyl) -N'phenylurea compound was performed by thin layer chromatography with 3 different solvents (hexan: ethyl acetate: methanol = 2: 3: 1; Hexan: acetone = 4: 2; Hexan: ethylacetate = 4: 2) a single stain is obtained which is different from the Rf value compared to the N-phenylurea origin compound. The Melting Point of the compound is 149oC so it is seen that the compound has been formed and different from the origin compound of N-phenylurea. The anticancer activity test after performed with MTT assay method using HeLa cells line, has an IC50 2100 mg / ml or 8,52 mM for the  N- (2-chlorobenzoyl) -N'phenylurea and higher than hydroxy urea as standard compound which has the activity as 7537 mg / ml or 99,10 mM.


 N- (2-chlorobenzoyl) -N'phenylurea compound has been successfully synthesized and can be further developed as an anticancer compound.


Synthesis; N-(2-chlorobenzoyl)-N'phenylurea; anticancer activity; HeLa cells.



Abraham, DJ. 2003. Burger's Medicinal Chemistry and Drug Discovery, Sixth Edition, Volume 1: Drug Discovery, John Wiley and Sons, Inc., New York.

Al-Amin M. and Rabiul, I.M., 2005. Cytotoxicity Study of Some Ketone Schiff Bases and Heterocyclic Compounds; Dhaka University Journal of Pharmaceutical Sciences, Vol.4, No.1.

Avendano C and Menendes J.C, 2008, Medicinal Chemistry of Anticancer Drugs, Elsevier, Amsterdam: 13-18

Bambang Tri Purwanto, 1991, Sintesis dan Uji Aktivitas Antibakteri N-para-bromobenzoilampisilin terhadap S.aureus ATCC 7500 dan E.coli ATCC 7500, Tesis Magister, Program Pascasarjana UGM.

Bambang Soekardjo, 1989, Sintesis dan hubungan struktur dengan aktivitas in vitro dari suatu seri turunan benzoil-N-ampisilin baru, Disertasi Doktor, Program Pascasarjana Universitas Airlangga.

Clayden, G., Warren, Wothers, 2001, Organic Chemistry, Oxford University Press, New York : 279-303;

Diyah, N.W., Purwanto, Siswandono, Hardjono, S., Purwanto, B.T., Susilowati, R. 2007. The Structure Modification And Analgesic Activities Of O-Benzoylsalicylic Acid Derivatives. USM–UNAIR First Collaborative Conference, Penang.

DepKes RI, 2006. Departemen Kesehatan Bentuk Subdirektorat Kanker, TEMPO Interaktif, Diakses pada 14 Maret 2010.

Fortin, S., Moreau, E., Patenaude, A., Desjardins, M. Lacroix, J., Rousseau, J.L.C., and Rene´ C-Gaudreaulta, R. 2005. Optimized N-phenyl-N0-(2-chloroethyl)ureas as potential antineoplastic agents: Synthesis and growth inhibition activity. Bioorganic & Medicinal Chemistry 13 : 6703–6712

Hardjono, 2011, Modifikasi Struktur 1-(Benzoiloksi)urea dan Hubungan Kuantitatif Struktur Aktivitas Sitotoksiknya. Disertasi. Fakultas Sains dan Teknologi Universitas Airlangga, Surabaya.
Kar, A. 2007. Medicinal Chemistry, 4th Ed., New Age International Ltd Publishers, New Delhi. pp. 794 – 810

Li, JN., Song, D., Lin, YH., Hu, QY., Yin, L., Bekesi, G., Holland, JF., Jiang, JD. 2003. Inhibition of Microtubule Polymerization by 3-Bromopropionylaminobenzoylurea (JIMB01), a New Cancericidal Tubulin Ligand. Biochemical Pharmacology 65 : 1691–1699.

McLaughlin JL, 1991, Crown Gall Tumour on Potato Discs and Brine Shrimp Lethality: Two Simple Bioassays for Higher Plant Screening and Fractionation, Academic Press Limited., London.

Mc Murry JM, 2008. Organic Chemistry. Belmont: Broke/Cole, pp 263-289; 779-851.

Moreau, E., Fortin, S., Lacroix, J., Patenaude, A. 2008. N-Phenyl-N0-(2-chloroethyl)ureas (CEUs) as potential antineoplastic agents. Part 3: Role of carbonyl groups in the covalent binding to the colchicine-binding site. Bioorganic & Medicinal Chemistry 16 : 1206–1217

Platt, O.S. 2008. Hydroxyurea for the treatment of sickle cell anemia. N. Engl. J. Med. 358 (13): 1362–9.

Reksohadiprodjo MS, 1981. Transformation of Fusel Oil from Indonesian Alcohol Factories into Products of Higher Value. Unpublished Doctor Dissertation, Gadjah Mada University, pp. 134, 145-152.

Saban, N. Bujak, M. 2009. Hydroxyurea and Hydroxamic Acid Derivatives as Antitumor Drugs. Cancer Chemother Pharmacol. 64 : 213 – 221

Silverstein R.M., Webster, F.X., and Kiemle D.J. 2005. Spectrofotometric Identification of Organic Compound, 7th Ed, John Wiley and Sons, Inc., New York.

Siswandono. 1999. Sintesis Senyawa Baru Turunan Asil Dan Benzoil-N-Urea Untuk Optimasi Aktivitas Penekan
Sistem Saraf Pusat. Laporan Penelitian Riset Unggulan Terpadu VI, Kementerian Ristek RI, LIPI., Jakarta.

Siswandono dan Soekardjo,B., 2000, Kimia Medisinal I & II, Airlangga University Press, Surabaya.
Susilowati, R., Diyah, NW., Sulistiawan, A., Lestari, Y.M. 2011. Aktivitas Sitotoksik Turunan N-Benzoilurea. Laporan Penelitian Fakultas Farmasi Unair, Surabaya.

Suzana, Diyah, N.W., Ekowati, J. 2009. Synthesis and CNS-Depressant Activity of Benzoylthiourea Derivatives. UNAIR–USM Second Collaborative Conference, Surabaya.

Tjiptasurasa, 1991. Komponen minyak fusel Indonesia, isolasi dan oksidasinya, sintesis dan evaluasi farmakokimia beberapa senyawa turunannya, Unpublished Doctor Dissertation, Universitas Gadjah Mada, Yogyakarta.

Taylor JB, Kennewell PD, 1993. Modern Medicinal Chemistry. Chichester : Ellis Horwood Limited, pp 272-275.

Zhukov, N.V. Tjulandin, S.A. 2008. Targeted therapy in the treatment of solid tumors: practice contradicts theory. Biochemistry Mosc. 73 (5): 605–618.
How to Cite
PURWANTO, Bambang Tri. Synthesis and Anti cancer Activity Test against HeLa Cells from N-(2-Chlorobenzoyl)-N’-Phenylurea. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 16, n. 2, p. 159-165, oct. 2018. ISSN 2614-6495. Available at: <>. Date accessed: 19 may 2024. doi: