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Table 1. Chemical characteristics and cytostatic effect of Pem- derivativesCompoundMS [M] a R t (min) b IC 50 (µM) cCalculated Measured NCI-H358 HL-60IELLQARGGC[Pem]-amide 1525.7 1525.8 28.7 3.53 2.17RRRRRRRRGGC[Pem]-amide1951.2 1951.5 24.4 7.70 6.24IELLQARGGC[Pem]-GGRRRRRRRR-amide2889.3 2890.3 33.3 1.93 2.64Pem 427.1 427.3 32.2 1.05 0.55Pem(OMe) 2 455.2 455.3 29.0 87.34 9.44a ESI-MS; b HPLC retention time, SupelcosilTM LC-18-DB (C18, 120 Å, 5µm, 4.6 × 250mm; Bellefonte, PA., U.S.A.) column, gradient elution: 0-5 min 5% B eluent, 5-50 min 95%B eluent, where eluent A: 0.1% TFA in water, eluent B: 0.1% TFA in ACN : water (80 : 20,v/v); c IC 50 values of compounds in µM on NCI-H358 and HL-60 cells.Compounds were purified with semipreparative RP-HPLC and chemically characterized byanalytical RP-HPLC and ESI-MS. For the analytical RP-HPLC SupelcosilTM LC-18-DB(C18, 120 Å, 5µm, 4.6 × 250 mm; Bellefonte, PA, U.S.A.) column was used, gradient: 0-5min 5% B eluent, 5-50 min 95% B eluent, where eluent A: 0.1% TFA in water, eluent B:0.1% TFA in ACN : water (80 : 20, v/v). Cytostatic effect was determined by MTT-assay[6] on HL-60 human leukemia and NCI-H358 human non-small cell lung carcinoma celllines. Table 1 shows the calculated and measured molecular mass, retention time and IC 50values of studied compounds.We found that attachment of Pem to three oligopeptides studied (IELLQARGGC,RRRRRRRRGGC, and IELLQARGGC-GGRRRRRRRR) only slightly decreased thecytostatic effect of free Pem in both cell lines (IC 50 changed from 1.05 to 7.7 in case ofNCI-H358 cell and from 0.55 to 6.24 in case of HL-60 cells). It is interesting to note thatthere was no significant difference between the cytostatic effect of Pem-conjugates on NCI-H358 and on HL-60 cells, Pem(OMe) 2 was not as effective as Pem with free carboxylicgroups.AcknowledgmentsThese studies were supported by grants from Hungarian Research Fund (OTKA, No. K68285),Hungarian Ministry of Education (NKFP 1A005/04, Medichem 2), and grants from the National Officefor Research and Technology, Hungary (3.2.1 – 2004-04-0005/3.0 and 3.2.1-2004-04-0352/3.0).References1. Hudecz, F., Reményi, J., Szabó, R., Kóczán, Gy., Mező, G., Kovács, P., Gaál, D. J. Mol.Recognition 16, 288-298 (2003).2. Miklán, Zs., Orbán, E., Csík, G., Schlosser, G., Magyar, A., Hudecz, F. Biopolymers PeptideScience 92, 489-501 (2009).3. Miklán, Zs., Szabó, R., Schlosser, G., Andreu, D., Hudecz, F., In Rolka, K., Rekowski, P. andSilberring, J. (Eds.) Peptides 2006: (Proceedings of the 29th European Peptide Symposium), KenesInternational, Switzerland, 2007, p. 538-539.4. Hudecz, F., Bánóczi, Z., Csík, G. Medicinal Research Reviews 25, 679-786 (2005).5. Fukuda, M., et al. Cancer Research 60, 450-456 (2000).6. Slater, T.F., Sawyer, B., Strauli, U. Biochimica Biophysica Acta, 77, 383-393 (1963).395