Genotoxic Impurities in Pharmaceutical Manufacturing: Sources ...
Genotoxic Impurities in Pharmaceutical Manufacturing: Sources, Regulations and Mitigation 1,
2
3
2,
Gyorgy Szekely *, Miriam C. Amores de Sousa , Marco Gil , Frederico Castelo Ferreira *, 3,
William Heggie *
1. School of Chemical Engineering & Analytical Science, The University of Manchester, The Mill, Sackville street, Manchester M13 9PL, United Kingdom 2. Department of Bioengineering and Institute for Bioengineering and Biosciences (iBB), Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001, Lisbon, Portugal 3. Hovione FarmaCiencia SA, R&D, Sete Casas, 2674-506, Loures, Portugal
Supporting Information – Table of Contents Complete list of authors for references 28, 34, 35, 48, 67, 74, 115, 123, 130, 131, 143, 145, 148, 172, 179, 181, 185, 196, 232, 234 and 289.
S1
(28) Muller, L.; Mauthe, R. J.; Riley, C. M.; Andino, M. M.; Antonis, D. D.; Beels, C.; DeGeorge, J; De Knaep, A. G.; Ellison, D.; Fagerland, J.A.; Frank, R.; Fritschel, B.; Galloway, S.; Harpur, E.; Humfrey, C. D.; Jacks, A. S.; Jagota, N.; Mackinnon, J.; Mohan, G.; Ness, D. K.; O'Donovan, M. R.; Smith, M. D.; Vudathala, G.; Yotti, L. A Rationale for Determining, Testing, and Controlling Specific Impurities in Pharmaceuticals that Possess Potential for Genotoxicity. Regul. Toxicol. Pharmacol. 2006, 44, 198-211.
(34) Ripin, D. H. B.; Teager, D. S.; Fortunak, J.; Basha, S. M.; Bivins, N.; Boddy, C.N.; Byrn, S.; Catlin, K. K.; Houghton, S. R.; Jagadeesh, S. T.; Kumar, K. A.; Melton, J.; Muneer, S.; Rao, L. N.; Rao, R. V.: Ray, P. C.; Reddy, N. G.; Reddy, R. M.; Shekar, K. C.; Silverton, T.; Smith, D. T.; Stringham, R. W.; Subbaraju, G.V.; Talley, F.; Williams, A. Process Improvements for the Manufacture of Tenofovir Disoproxil Fumarate at Commercial Scale. Org. Process Res. Dev. 2010, 14, 1194-1201.
(35) Dunetz, J. R.; Berliner, M. A.; Xiang, Y.; Houck, T. L.; Salingue, F. H.; Chao, W.; Yuandong, C.; Shenghua, W.; Huang, Y.; Farrand, D.; Boucher, S. J.; Damon, D. B.; Makowski, T. W.; Barrila, M. T.; Chen, R.; Martínez, I. Multikilogram Synthesis of a Hepatoselective Glucokinase Activator. Org. Process Res. Dev. 2012, 16, 1635-1645.
(48) Yang, Q.; Haney, B. P.; Vaux, A.; Riley, D. A.; Heidrich, L.; He, P.; Mason, P.; Tehim, A.; Fisher, L. E.; Maag, H.; Anderson, N. G. Controlling the Genotoxins Ethyl Chloride and Methyl Chloride Formed During the Preparation of Amine Hydrochloride Salts from Solutions of Ethanol and Methanol. Org. Process Res. Dev. 2009, 13, 786-791.
(67) Mu, J.; Petrov, A.; Eiermann, G. J.; Woods, J.; Zhou, Y.P.; Li, Z.; Zycband, E.; Feng, Y.; Zhu, L.; Roy, R. S.; Howard, A. D.; Li, C.; Thornberry, N. A.; Zhang, B. B. Inhibition of DPP-4 with Sitagliptin Improves Glycemic Control and Restores Islet Cell Mass and Function in a Rodent Model of Type 2 Diabetes. Eur. J. Pharmacol. 2009, 623, 148-154.
(74) Harris, P. A.; Boloor, A.; Cheung, M.; Kumar, R.; Crosby, R. M.; Davis-Ward, R. G.; Epperly, A. H.; Hinkle, K. W.; Hunter, R. N.; Johnson, J. H.; Knick, V. B.; Laudeman, C. P.; Luttrell, D. K.; Mook, R. A.; Nolte, R. T.; Rudolph, S. K.; Szewczyk, J. R.; Truesdale, A. T.; Veal, J. M.; Wang, L.; Stafford J. A. Discovery of 5-[[4-[(2,3-Dimethyl-2H-indazol-6yl)methylamino]-2-pyrimidinyl]amino]-2-methyl-benzenesulfonamide (Pazopanib), a Novel and Potent Vascular Endothelial Growth Factor Receptor Inhibitor. J. Med. Chem. 2008, 51, 4632-4640.
(115) Sun, L.; Liang, C.; Shirazian, S.; Zhou, Y.; Miller, T.; Cui, J.; Fukuda, J. Y.; Chu, J. Y.; Nematalla, A.; Wang, X.; Chen, H.; Sistla, A.; Luu, T. C.; Tang, F.; Wei, J.; Tang, C. Discovery of 5-[5-Fluoro-2-oxo-1,2- dihydroindol-(3Z)ylidenemethyl]-2,4- dimethyl-1H-pyrrole-3-carboxylic Acid (2-Diethylaminoethyl)amide, a Novel Tyrosine Kinase Inhibitor Targeting Vascular Endothelial and Platelet-Derived Growth Factor Receptor Tyrosine Kinase. J. Med. Chem. 2003, 46, 1116.
S2
(123)
Parkes, K. E. B., Bushnell, D. J., Crackett, P. H., Dunson, S. J.; Freeman, A. C.; Gunn, H. P.; Hopkins, R. A.;
Lambert, R. W.; Martin, J. A.; Merret, J. H.; Redshaws, S.; Spurden, W. C.; Thomas, G. T. Studies toward the LargeScale Synthesis of the HIV Proteinase Inhibitor Ro 31-8959. J. Org. Chem. 1994, 59, 3656-3654.
(130)
Showalter, H. D. H.; Angelo, M. M.; Berman, E. M.; Kanter, G. D.; Ortwine, D. F.; Ross-Kesten, S. G.; Sercel,
A. D.; Turner, W. R.; Werbel, L. M.; Worth, D. F.; Elslager, E. F.; Leopold, W. R.; Shillis, J. L. Benzothiopyranoindazoles, a New Class of Chromophore Modified Anthracenedione Anticancer Agents. Synthesis and Activity Against Murine Leukemias. J. Med. Chem. 1988, 31, 1527-1539.
(131)
Beylin, B. V.; Colbry, N. L.; Giordani, A. B.; Goel, O. P.; Jonhson, D. M.; Leeds, R. L.; Leja, B.; Lewis, E. P.;
Lustgarten, D. M.; Showalter, H. D. H; Sercel, A. D.; Reily, M. D; Uhlendorf, S. E.; Zisek, K. A.; Mcdonnell, P. An Improved Synthesis of Anticancer Benzothiopyranoindazoles. An Efficient Large-Scale β-Aminoethylation Procedure. J. Heterocycl. Chem. 1991, 28, 517-527.
(143)
Fei, Z.; Wu, Q.; Zhang, F.; Cao, Y.; Liu, C.; Shieh, W. C.; Xue, S.; McKenna, J.; Prasad, K.; Prashad, M.;
Baeschlin, D.; Namoto, K. A Scalable Synthesis of an Azabicyclooctanyl Derivative, a Novel DPP-4 Inhibitor. J. Org. Chem. 2008, 73, 9016-9021.
(145)
Kumar, S.; Dare, L.; Vasko-Moser, J. A.; James, I. E.; Blake, S. M.; Rickard, D. J.; Hwang, S. M.; Tomaszek,
T.; Yamashita, D. S.; Marquis, R. W.; Oh, H.; Jeong, J. U.; Veber, D. F.; Gowen, M.; Lark, M. W.; Stroup, G. A Highly Potent Inhibitor of Cathepsin K (Relacatib) Reduces Biomarkers of Bone Resorption both In Vitro and in an Acute Model of Elevated Bone Turnover In Vivo in Monkeys. Bone 2007, 40, 122-131.
(148)
George, D.T.; Gilman, J.; Hersh, J.; Thorsell, A.; Herion, D.; Geyer, C.; Peng, X.; Kielbasa, W.; Rawlings, R.;
Brandt, J. E.; Gehlert, D. R.; Tauscher, J. T.; Hunt, S. P.; Hommer, D.; Heilig, M. Neurokinin 1 Receptor Antagonism as a Possible Therapy for Alcoholism. Science 2008, 319, 1536-1539.
(172)
Larsen, R. D.; King, A. O.; Chen, C. Y.; Corley, E. G.; Foster, B. S.; Roberts, F. E.; Yang, C.; Lieberman, D.
R.; Reamer, R. A.; Tschaen, D. M.; Verhoeven, T. R.; Reider, P. J.; Lo, Y. S.; Rossano, L. T.; Brookes, A. S.; Meloni, D.; Moore, J. R.; Arnett, J. F. Efficient Synthesis of Losartan, A Nonpeptide Angiotensin II Receptor Antagonist. J. Org. Chem. 1994, 59, 6391-6394.
(179)
Teasdale, A.; Eyley, S. C.; Delaney, E.; Jacq, K.; Taylor-Worth, K.; Lipczynski, A.; Reif, V.; Elder, D. P.;
Facchine, K. L.; Golec, S.; Oestrich, R.S.; Sandra, P.; David, F. Mechanism and Processing Parameters Affecting the Formation of Methyl Methanesulfonate from Methanol and Methanesulfonic Acid: An Illustrative Example for Sulfonate Ester Impurity Formation. Org. Process Res. Dev. 2009, 13, 429-433.
S3
(181)
Dressman, B. A.; Fritz, J. E.; Hammond, M.; Hornback, W. J.; Kaldor, S. W.; Kalish, V. J.; Munroe, J. E.;
Reich, S. H.; Tatlock, J. H.; Shepherd, T. A.; Rodriguez, M. J. HIV Protease Inhibitors. U.S. Patent 5,484,926, January 16, 1996.
(185)
Teasdale, A.; Delaney, E.; Eyley, S. C.; Jacq, K.; Taylor-Worth, K.; Lipczynski, A.; Hoffmann, W.; Reif, V.;
Elder, D. P.; Facchine, K. L.; Golec, S.; Oestrich, R.S.; Sandra, P.; David, F. A Detailed Study of Sulfonate Ester Formation and Solvolysis Reaction Rates and Application toward Establishing Sulfonate Ester Control in Pharmaceutical Manufacturing Processes. Org. Process Res. Dev. 2010, 14, 999-1007.
(196)
Brickner, S. J.; Hutchinson, D. K.; Barbachyn, M. R.; Manninen, P. R.; Ulanowicz, D. A.; Garmon, S. A.;
Grega, K. C.; Hendges, S. K.; Toops, D. S.; Foed, C. W.; Zurenko, G. E. Synthesis and Antibacterial Activity of U100592 and U-100766, Two Oxazolidinone Antibacterial Agents for the Potential Treatment of Multidrug-Resistant Gram-Positive Bacterial Infections. J. Med. Chem. 1996, 39, 673-679.
(232)
Stuk, T. L.; Allen, M. S.; Haight, A. R.; Kerdesky, F. A.; Langridge, D. C.; Leanna, R. M.; Lijewski, L. M.;
Melcher, L.; Morton, H. E.; Norbeck, D. W.; Reno, D. S.; Robbins, T. A.; Scarpetti, D.; Sham, H. L.; Sowin, T. J.; Tien, J. J.; Zhao, C. Process for the Preparation of a Substituted 2,5-Diamino-3-hydroxyhexane. U.S. Patent 5,491,253, February 13, 1996.
(234)
Stimac, A.; Mohar, B.; Stephan, M.; Bevc, M.; Zupet, R.; Gartner, A.; Kroselj, V.; Smrkolj, M.; Kidemet, D.;
Sedmak, G.; Benkic, P.; Kljajic, A.; Plevnik, M. Process for the Preparation of Ezetimibe and Derivatives Thereof. Int. Patent WO2008089984, September 18, 2008.
(289)
Welch, C. J.; Albaneze-Walker, J.; Leonard, W. R.; Biba, M.; DaSilva, J.; Henderson, D.; Laing, B.; Mathre,
D. J.; Spencer, S.; Bu, X.; Wang, T. Adsorbent Screening for Metal Impurity Removal in Pharmaceutical Process Research. Org. Process Res. Dev. 2005, 9, 198-205.
S4
2
3
2,
Gyorgy Szekely *, Miriam C. Amores de Sousa , Marco Gil , Frederico Castelo Ferreira *, 3,
William Heggie *
1. School of Chemical Engineering & Analytical Science, The University of Manchester, The Mill, Sackville street, Manchester M13 9PL, United Kingdom 2. Department of Bioengineering and Institute for Bioengineering and Biosciences (iBB), Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001, Lisbon, Portugal 3. Hovione FarmaCiencia SA, R&D, Sete Casas, 2674-506, Loures, Portugal
Supporting Information – Table of Contents Complete list of authors for references 28, 34, 35, 48, 67, 74, 115, 123, 130, 131, 143, 145, 148, 172, 179, 181, 185, 196, 232, 234 and 289.
S1
(28) Muller, L.; Mauthe, R. J.; Riley, C. M.; Andino, M. M.; Antonis, D. D.; Beels, C.; DeGeorge, J; De Knaep, A. G.; Ellison, D.; Fagerland, J.A.; Frank, R.; Fritschel, B.; Galloway, S.; Harpur, E.; Humfrey, C. D.; Jacks, A. S.; Jagota, N.; Mackinnon, J.; Mohan, G.; Ness, D. K.; O'Donovan, M. R.; Smith, M. D.; Vudathala, G.; Yotti, L. A Rationale for Determining, Testing, and Controlling Specific Impurities in Pharmaceuticals that Possess Potential for Genotoxicity. Regul. Toxicol. Pharmacol. 2006, 44, 198-211.
(34) Ripin, D. H. B.; Teager, D. S.; Fortunak, J.; Basha, S. M.; Bivins, N.; Boddy, C.N.; Byrn, S.; Catlin, K. K.; Houghton, S. R.; Jagadeesh, S. T.; Kumar, K. A.; Melton, J.; Muneer, S.; Rao, L. N.; Rao, R. V.: Ray, P. C.; Reddy, N. G.; Reddy, R. M.; Shekar, K. C.; Silverton, T.; Smith, D. T.; Stringham, R. W.; Subbaraju, G.V.; Talley, F.; Williams, A. Process Improvements for the Manufacture of Tenofovir Disoproxil Fumarate at Commercial Scale. Org. Process Res. Dev. 2010, 14, 1194-1201.
(35) Dunetz, J. R.; Berliner, M. A.; Xiang, Y.; Houck, T. L.; Salingue, F. H.; Chao, W.; Yuandong, C.; Shenghua, W.; Huang, Y.; Farrand, D.; Boucher, S. J.; Damon, D. B.; Makowski, T. W.; Barrila, M. T.; Chen, R.; Martínez, I. Multikilogram Synthesis of a Hepatoselective Glucokinase Activator. Org. Process Res. Dev. 2012, 16, 1635-1645.
(48) Yang, Q.; Haney, B. P.; Vaux, A.; Riley, D. A.; Heidrich, L.; He, P.; Mason, P.; Tehim, A.; Fisher, L. E.; Maag, H.; Anderson, N. G. Controlling the Genotoxins Ethyl Chloride and Methyl Chloride Formed During the Preparation of Amine Hydrochloride Salts from Solutions of Ethanol and Methanol. Org. Process Res. Dev. 2009, 13, 786-791.
(67) Mu, J.; Petrov, A.; Eiermann, G. J.; Woods, J.; Zhou, Y.P.; Li, Z.; Zycband, E.; Feng, Y.; Zhu, L.; Roy, R. S.; Howard, A. D.; Li, C.; Thornberry, N. A.; Zhang, B. B. Inhibition of DPP-4 with Sitagliptin Improves Glycemic Control and Restores Islet Cell Mass and Function in a Rodent Model of Type 2 Diabetes. Eur. J. Pharmacol. 2009, 623, 148-154.
(74) Harris, P. A.; Boloor, A.; Cheung, M.; Kumar, R.; Crosby, R. M.; Davis-Ward, R. G.; Epperly, A. H.; Hinkle, K. W.; Hunter, R. N.; Johnson, J. H.; Knick, V. B.; Laudeman, C. P.; Luttrell, D. K.; Mook, R. A.; Nolte, R. T.; Rudolph, S. K.; Szewczyk, J. R.; Truesdale, A. T.; Veal, J. M.; Wang, L.; Stafford J. A. Discovery of 5-[[4-[(2,3-Dimethyl-2H-indazol-6yl)methylamino]-2-pyrimidinyl]amino]-2-methyl-benzenesulfonamide (Pazopanib), a Novel and Potent Vascular Endothelial Growth Factor Receptor Inhibitor. J. Med. Chem. 2008, 51, 4632-4640.
(115) Sun, L.; Liang, C.; Shirazian, S.; Zhou, Y.; Miller, T.; Cui, J.; Fukuda, J. Y.; Chu, J. Y.; Nematalla, A.; Wang, X.; Chen, H.; Sistla, A.; Luu, T. C.; Tang, F.; Wei, J.; Tang, C. Discovery of 5-[5-Fluoro-2-oxo-1,2- dihydroindol-(3Z)ylidenemethyl]-2,4- dimethyl-1H-pyrrole-3-carboxylic Acid (2-Diethylaminoethyl)amide, a Novel Tyrosine Kinase Inhibitor Targeting Vascular Endothelial and Platelet-Derived Growth Factor Receptor Tyrosine Kinase. J. Med. Chem. 2003, 46, 1116.
S2
(123)
Parkes, K. E. B., Bushnell, D. J., Crackett, P. H., Dunson, S. J.; Freeman, A. C.; Gunn, H. P.; Hopkins, R. A.;
Lambert, R. W.; Martin, J. A.; Merret, J. H.; Redshaws, S.; Spurden, W. C.; Thomas, G. T. Studies toward the LargeScale Synthesis of the HIV Proteinase Inhibitor Ro 31-8959. J. Org. Chem. 1994, 59, 3656-3654.
(130)
Showalter, H. D. H.; Angelo, M. M.; Berman, E. M.; Kanter, G. D.; Ortwine, D. F.; Ross-Kesten, S. G.; Sercel,
A. D.; Turner, W. R.; Werbel, L. M.; Worth, D. F.; Elslager, E. F.; Leopold, W. R.; Shillis, J. L. Benzothiopyranoindazoles, a New Class of Chromophore Modified Anthracenedione Anticancer Agents. Synthesis and Activity Against Murine Leukemias. J. Med. Chem. 1988, 31, 1527-1539.
(131)
Beylin, B. V.; Colbry, N. L.; Giordani, A. B.; Goel, O. P.; Jonhson, D. M.; Leeds, R. L.; Leja, B.; Lewis, E. P.;
Lustgarten, D. M.; Showalter, H. D. H; Sercel, A. D.; Reily, M. D; Uhlendorf, S. E.; Zisek, K. A.; Mcdonnell, P. An Improved Synthesis of Anticancer Benzothiopyranoindazoles. An Efficient Large-Scale β-Aminoethylation Procedure. J. Heterocycl. Chem. 1991, 28, 517-527.
(143)
Fei, Z.; Wu, Q.; Zhang, F.; Cao, Y.; Liu, C.; Shieh, W. C.; Xue, S.; McKenna, J.; Prasad, K.; Prashad, M.;
Baeschlin, D.; Namoto, K. A Scalable Synthesis of an Azabicyclooctanyl Derivative, a Novel DPP-4 Inhibitor. J. Org. Chem. 2008, 73, 9016-9021.
(145)
Kumar, S.; Dare, L.; Vasko-Moser, J. A.; James, I. E.; Blake, S. M.; Rickard, D. J.; Hwang, S. M.; Tomaszek,
T.; Yamashita, D. S.; Marquis, R. W.; Oh, H.; Jeong, J. U.; Veber, D. F.; Gowen, M.; Lark, M. W.; Stroup, G. A Highly Potent Inhibitor of Cathepsin K (Relacatib) Reduces Biomarkers of Bone Resorption both In Vitro and in an Acute Model of Elevated Bone Turnover In Vivo in Monkeys. Bone 2007, 40, 122-131.
(148)
George, D.T.; Gilman, J.; Hersh, J.; Thorsell, A.; Herion, D.; Geyer, C.; Peng, X.; Kielbasa, W.; Rawlings, R.;
Brandt, J. E.; Gehlert, D. R.; Tauscher, J. T.; Hunt, S. P.; Hommer, D.; Heilig, M. Neurokinin 1 Receptor Antagonism as a Possible Therapy for Alcoholism. Science 2008, 319, 1536-1539.
(172)
Larsen, R. D.; King, A. O.; Chen, C. Y.; Corley, E. G.; Foster, B. S.; Roberts, F. E.; Yang, C.; Lieberman, D.
R.; Reamer, R. A.; Tschaen, D. M.; Verhoeven, T. R.; Reider, P. J.; Lo, Y. S.; Rossano, L. T.; Brookes, A. S.; Meloni, D.; Moore, J. R.; Arnett, J. F. Efficient Synthesis of Losartan, A Nonpeptide Angiotensin II Receptor Antagonist. J. Org. Chem. 1994, 59, 6391-6394.
(179)
Teasdale, A.; Eyley, S. C.; Delaney, E.; Jacq, K.; Taylor-Worth, K.; Lipczynski, A.; Reif, V.; Elder, D. P.;
Facchine, K. L.; Golec, S.; Oestrich, R.S.; Sandra, P.; David, F. Mechanism and Processing Parameters Affecting the Formation of Methyl Methanesulfonate from Methanol and Methanesulfonic Acid: An Illustrative Example for Sulfonate Ester Impurity Formation. Org. Process Res. Dev. 2009, 13, 429-433.
S3
(181)
Dressman, B. A.; Fritz, J. E.; Hammond, M.; Hornback, W. J.; Kaldor, S. W.; Kalish, V. J.; Munroe, J. E.;
Reich, S. H.; Tatlock, J. H.; Shepherd, T. A.; Rodriguez, M. J. HIV Protease Inhibitors. U.S. Patent 5,484,926, January 16, 1996.
(185)
Teasdale, A.; Delaney, E.; Eyley, S. C.; Jacq, K.; Taylor-Worth, K.; Lipczynski, A.; Hoffmann, W.; Reif, V.;
Elder, D. P.; Facchine, K. L.; Golec, S.; Oestrich, R.S.; Sandra, P.; David, F. A Detailed Study of Sulfonate Ester Formation and Solvolysis Reaction Rates and Application toward Establishing Sulfonate Ester Control in Pharmaceutical Manufacturing Processes. Org. Process Res. Dev. 2010, 14, 999-1007.
(196)
Brickner, S. J.; Hutchinson, D. K.; Barbachyn, M. R.; Manninen, P. R.; Ulanowicz, D. A.; Garmon, S. A.;
Grega, K. C.; Hendges, S. K.; Toops, D. S.; Foed, C. W.; Zurenko, G. E. Synthesis and Antibacterial Activity of U100592 and U-100766, Two Oxazolidinone Antibacterial Agents for the Potential Treatment of Multidrug-Resistant Gram-Positive Bacterial Infections. J. Med. Chem. 1996, 39, 673-679.
(232)
Stuk, T. L.; Allen, M. S.; Haight, A. R.; Kerdesky, F. A.; Langridge, D. C.; Leanna, R. M.; Lijewski, L. M.;
Melcher, L.; Morton, H. E.; Norbeck, D. W.; Reno, D. S.; Robbins, T. A.; Scarpetti, D.; Sham, H. L.; Sowin, T. J.; Tien, J. J.; Zhao, C. Process for the Preparation of a Substituted 2,5-Diamino-3-hydroxyhexane. U.S. Patent 5,491,253, February 13, 1996.
(234)
Stimac, A.; Mohar, B.; Stephan, M.; Bevc, M.; Zupet, R.; Gartner, A.; Kroselj, V.; Smrkolj, M.; Kidemet, D.;
Sedmak, G.; Benkic, P.; Kljajic, A.; Plevnik, M. Process for the Preparation of Ezetimibe and Derivatives Thereof. Int. Patent WO2008089984, September 18, 2008.
(289)
Welch, C. J.; Albaneze-Walker, J.; Leonard, W. R.; Biba, M.; DaSilva, J.; Henderson, D.; Laing, B.; Mathre,
D. J.; Spencer, S.; Bu, X.; Wang, T. Adsorbent Screening for Metal Impurity Removal in Pharmaceutical Process Research. Org. Process Res. Dev. 2005, 9, 198-205.
S4
