Tuberculostatic Agents
193
Copolymer Poly(ethylene glycol)-Poly(aspartic acid) (PEG-PASP) [25,30]
[7] World Health Organization; Bull. WHO 1992, 70, 17.
[8] H. Bundgaard, Ed.; Design of Prodrugs, Elsevier; Amsterdam, 1985.
A solution of 34.5 mL of NaOH 0.43 M in water-methanol-isopropyl
alcohol (1:2:2) was added to a solution of PEG-PBLA (2.7 g) in acetic acid
(27 mL) and the resultant was added to ethyl ether (40 mL). The precipitate
was dissolved in distilled water and dialysed against distilled water in a
dialysis membrane (cut off 1,000) for 4 hours. The solution was then
lyophilised. 1H-NMR (D2O): δ = 4.37(s, CHa- and b-amide), 3.43 (m,
CH2PEG), 3.19 (s, CH3), 2.77 (sl, CH2 a-amide). 13C-NMR (D2O): δ =
172.26 (C7,C11), 163.32 (C4), 69.98 (C2), 51.13 (C10), 35.06 (C3). Free
COOH: 0.595 mEq/g polymer.
[9] G. J. Friis, H. Bundgaard in A Textbook of Drug Design and Develop-
ment (Ed.: P. Krogsgaard-Larsen, T. Liljefors, U. Madsen) 2nd ed
Harwood Academic Press, Oxford, 1996, chapter 13.
[10] C. G. Wermuth in The practice of medicinal chemistry (Ed.: C. G.
Wermuth) Academic Press, New York, 1996.
[11] S. Cammas, K. Kataoka, Makromol. Chem. Phys. 1995, 196, 1899.
[12] G. Giammona, G. Cavallaro, G. Pitarresi, C. Ventura, S. Palazzo, Int.
J. Pharm. 1994, 105, 57.
Isoniazid Conjugate-Copolymer Poly(ethylene glycol)-Poly(aspartic acid)
(PEG-PASP-INH) [25,30]
[13] C. M. Samour in Polymeric drugs (Ed.: L.G. Donaruma, O. Vogl)
Academic Press, New York, 1978.
Isoniazid (0.18 g; 0.0013 mol) was dissolved in DMF (0.8 mL), then added
to a solution of EDC (0.52 g; 0.0033 mol) and PEG-PASP (1.51 g) in distilled
water (48.3 mL), the mixture was stirred for 4 hours at 0 °C. EDC (0.52 g;
0.0033 mol) was then added and the stirred mixture maintained for 24 hours
at room temperature. The solution obtained was dialysed against acetate
buffer pH 5.0 (dialysis tubing cut off 1,000), for 4 hours, against water,
dialysed for an additional 4 hours (dialysis tubing cut off 12,000–14,000),
then lyophilised. 1H-NMR (D2O, d): 8.13 (sl, NH polymer), 7.8–7.3 (dl,
CHINH), 4.84 (s, NH-NH), 4.62 (sl, CHα- and β-amide), 3.65 (m. CH2PEG),
3.36(s, OCH3), 2.11 (sl, CH2 α-amide). 13C-NMR (D2O, ppm): 176.42 (C7),
172.26 (C12), 165.27 (C4,C8), 150.09(C16,C17), 122.36 (C14,C15), 68.98
(C2), 58.79(C1), 39.51(C3), 34.89(C6). Free COOH: 0.209 mEq/g polymer.
[14] M. Shikawa, A. Kamijo, T. Fujita, et al., Pharm. Res. 1993, 10, 1253.
[15] H. Sezaki, Y. Takakura, M. Hashida, Adv. Drug Delivery Rev. 1989, 3,
247.
[16] C. J. T. Hoes, W. Potman, W. A. Von Heeswijk, et al., J. Control. Rel.
1985, 2, 205.
[17] L.W Seymour, K. Ulbrich, J. Strohalm, et al., Biochem. Pharmacol.
1990, 39, 1125.
[18] Y. Takakura, M. Hashida, Crit. Rev. Oncol. Hematol. 1994, 18, 207.
[19] A. A.Sinkula in Design of prodrugs (Ed.: H. Bundgaard) Elsevier,
Amsterdam, 1985, chapter 4.
[20] M. Yokoyama, M. Miyauchi, N. Yamada, et al., Cancer Res. 1990, 50,
In Vitro Biological Assay
1693.
MIC determination was carried out in autoclaved (121 °C/15 min) Mid-
dlebrook 7H9 culture medium (4.7 g) mixed with sterile water (900 mL) and
glycerol (2 mL). After cooling, 20 mL of OADC (oleic acid, albumin,
dextrose and catalase) was added for each 180 mL of culture medium.
Isoniazid and a negative control were compared to the synthesised polymers,
PEG-BLA AND PEG-PASP, in 8 twofold-concentrations each, from 3.2 to
0.025 mg/mL, using 8 mL of culture medium. Plates with 96 wells and
200 mL of culture medium/well were used for µMIC determination.
M. tuberculosis H37Rv strain was used in growth logarithm phase.
[21] J. Cassidy, R. Duncan, G. J. Morrison et al., Biochem. Pharmacol. 1989,
38, 875.
[22] G. Pratesi, G. Savi, G. Pezzoni et al., Br. J. Cancer 1985, 52, 841.
[23] W. R. Sorenson, T. W. Campbell, Preparative methods of polymer,
Interscience Publishers, New York, 1962.
[24] A. Zaffaroni, P. Bonsen in Polymeric Drugs (Ed.: L. G. Donaruma, O.
Vogel) Academic Press, New York, 1978, chapter 1.
[25] M. Yokoyama, M. Miyauchi, N. Yamada et al. in Advances in drug
delivery systems, 4. (Ed.: J. M. Anderson, S. W. Kim, K. Knutson)
Elsevier; Amsterdam, 1987.
References
[1] World Health Organization; Global Tuberculosis Control; WHO An-
nual Report 2000, Geneva. Available in the Internet.
[26] G. Kwon, M. Naito, M. Yokoyama, T. Okano et al., Langmuir 1993, 9,
945.
[27] M. Yokoyama, T. Okano, Y. Sakurai et al. Cancer Res. 1991, 51, 3229.
[28] L. A. Benoiton, Can. J. Chem. 1962, 40, 570.
[2] World Health Organization; Anti-tuberculosis Drug-resistant in the
World; Geneva, 1998.
[3] A. Korokolvas, Essentials of Medicinal Chemistry. 2nd. Ed., Wiley-In-
tersciense; New York, 1988.
[29] R. H. Karlson, K. S. Norland, G. D. Fasman, E. R. Bluot,J. Am. Chem.
Soc. 1960, 82, 2268.
[4] A. Korolkovas, Dicionário Terapêutico Guanabara 2000/2001. 6ª Ed.,
Guanabara Koogan, Rio de Janeiro, 2000.
[30] M. Yokoyama, S. Inoue, K. Kataoka, N. Yui, Y. Sakurai, Makromol.
Chem. Rapid Commun. 1987, 8, 431.
[5] P. Godfrey-Faussett, Policy statement on preventive therapy against
tuberculosis in people living with HIV, 1999 Available in the Internet.
[31] W. D. Fuller, M. S. Verlander, M. Goodman, Biopolymers 1976, 15,
1869.
[32] P. C. Hiemenz, Principles of Colloid and Surface Chemistry, 2nd ed.,
Marcel Dekker, New York, 1986.
[6] World Health Organization; WHO tuberculosis site. Dots: Directly
Observed Treatment Short-course, 2000. Available in the Internet
Received: March 14, 2000 [FP467]
Arch. Pharm. Pharm. Med. Chem. 334, 189–193 (2001)