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European Journal of Medicinal Chemistry 45 (2010) 5827e5832

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European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech

Original article

The synthesis of phenylalanine-derived C5-substituted rhodanines and their activity against selected methicillin-resistant Staphylococcus aureus (MRSA) strains
Diane Hardej, Charles R. Ashby Jr., Nikhil S. Khadtare, Shridhar S. Kulkarni, Satyakam Singh, Tanaji T. Talele*
Department of Pharmaceutical Sciences, College of Pharmacy and Allied Health Professions, St. John’s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA

a r t i c l e i n f o
Article history: Received 26 April 2010 Received in revised form 14 September 2010 Accepted 20 September 2010 Available online 25 September 2010 Keywords: MRSA Rhodanine Serum

a b s t r a c t
A series of rhodanine compounds containing various substituents at the N3- and C5-positions were synthesized and their in vitro activity against a panel of clinically relevant MRSA strains was determined. The anti-MRSA activity of compounds 21 (MIC ¼ 3.9 mg/mL, MBC ¼ 7.8 mg/mL) and 22 (MIC ¼ 1.95 mg/mL, MBC ¼ 7.8 mg/mL) was significantly greater than that of the lead compounds, 1e3 and reference antibiotics penicillin G (MIC ¼ 31.25 mg/mL) and ciprofloxacin (MIC ¼ 7.8 mg/mL) and comparable to that of vancomycin (MIC ¼ 0.97 mg/mL). Compounds 21 and 22 were found to be bactericidal at only 2e4-fold higher than their MIC concentrations. In addition, their MIC values remained unchanged in the presence or absence of 10% serum. Overall, the results suggest that compounds 21 and 22 may be of potential use in the treatment of MRSA infections. Ó 2010 Elsevier Masson SAS. All rights reserved.

1. Introduction Methicillin-resistant Staphylococcus aureus (MRSA) has become endemic in most hospitals and health care facilities in Western nations. In the United States, MRSA infections cause an estimated 19,000 deaths per annum [1]. The two major types, hospital (HA)and community-acquired (CA) MRSA, are significantly different in their epidemiological and genetic characteristics [2e4]. HA-MRSA occurs in patients with specific risk factors [5e7], whereas CA-MRSA can occur in healthy individuals that do not have predisposing factors [8e10]. Typically, HA-MRSA infections occur in the urine, lungs, bloodstream and surgical sites whereas CA-MRSA primarily produces skin and skin structure infections [4,11]. The HA-MRSA strains contain the mobile class I, II and III staphylococcus chromosome cassettes mec (SCCmec) and resistance to the b-lactam antibiotics is due to the encoding of the penicillin binding protein (PBP) 2a by the mecA gene [12e14]. In contrast, the resistance to non-b-lactams is due to the presence of transposons and other genes. PBP2a has an elevated rate constant for dissociation and decreased acylation rate constant and these together significantly decrease the acylation of PBP2a, producing b-lactam resistance [15]. This, in combination with

* Corresponding author. Tel.: þ1 718 990 5405; fax: þ1 718 990 1877. E-mail address: talelet@stjohns.edu (T.T. Talele). 0223-5234/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved. doi:10.1016/j.ejmech.2010.09.045

other genes, has allowed MRSA to become resistant to many different classes of antibiotics [16,17]. In contrast, CA-MRSA strains harbor SCCmec IV [18], a smaller and more mobile SCCmec than classes IeIII and this may contribute to its widespread dissemination [19,20]. The CA-MRSA strains, like HA-MRSA, are broadly resistant to b-lactam and macrolide/azalide antimicrobials but responsive to certain nonb-lactam antibiotics [3,21]. However, resistance rates are increasing and there are other limitations in the use of these drugs. Thus, given the widespread dissemination and mortality caused by MRSA, the synthesis and development of new drugs is imperative. Rhodanine-based molecules have been shown to be small molecule inhibitors of numerous targets such as hepatitis C virus (HCV) NS3 protease [22], aldose reductase [23,24], b-lactamase [25], UDP-Nacetylmuramate/L-alanine ligase [26], penicillin binding protein (PBP) [27], antidiabetic agents [28], cathepsin D [29] and histidine decarboxylase [30]. In addition, we have reported that rhodanine analogs are inhibitors of the HCV enzyme NS5B polymerase [31]. In a search for new antibacterial compounds from our in-house rhodanine library, we initially identified three rhodanine derivatives [1: MIC ¼ 62.5 mg/mL; 2: MIC ¼ 250 mg/mL and 3: MIC ¼ 125 mg/mL] exhibiting modest activity against selected MRSA strains (Chart 1). Here, we report the subsequent synthesis, anti-MRSA activity and SAR optimization focusing on N3- and C5-positions of the rhodanine core of these lead compounds. A variety of strains of S. aureus were used for this study and were obtained from the American Type Tissue

In contrast.6 mg/mL) as described below.5 mg/mL were as active as lead compound 1.5 as a singlet (except cinnamylidene analog 16 which showed a doublet) whereas the methine proton of the Z isomer of cyclohexylmethylidene analog (18) appeared at 6. MIC ¼ 250 mg/mL). there is a paucity of information about the contribution of the amino acid side chain at N3-position and uniquely substituted arylidenes at C5-position to their antibacterial action [25. Hardej et al. 700787. The Knoevenagel condensation reaction with aromatic aldehydes provided only the Z isomer.6 mg/mL). 2e4-fold improvement in inhibitory activity was observed in the presence of the 2. MIC ¼ 15. 4-fluorobenzylidene (compound 6) and naphthyledene-1-yl (compound 7) analogs each exhibiting MIC value of 62. Based on the SAR information gathered. Synthesis of compounds 14–18 and 20–22. MIC ¼ 15. Based on these findings. MIC ¼ 250 mg/ mL) and styryl (compound 16.7 to 8. the less bulky a-methylacetic acid analog (compound 13. a-methylthioethylacetic acid (compound 10. focusing on variations at the C5-benzylidene substituent. whereas compounds 14e18 and 20e22 were synthesized and tested for their in vitro activity against a panel of MRSA strains together with reference antibiotics ciprofloxacin. Although the SAR of rhodanine derivatives has been studied extensively. Strains 700698.36]. Strain BAA-1680 represents a community-acquired MRSA (CA-MRSA) strain from the skin of a patient from Michigan. ATCC strain 34404 is commonly used as a quality control organism for susceptibility testing. 2.. lead compounds 1e3. respectively). blood culture from New York and nasal culture from Hungary. / European Journal of Medicinal Chemistry 45 (2010) 5827e5832 Chart 1.S. commercially available rhodanine analogs (compounds 5e13) were obtained for exploration of the SAR around the N3. This downfield movement of methine proton in (Z) C5-benzylidene and alkylmethylidene analogs was reported to be due to the deshielding effect of the adjacent carbonyl group [28. MIC ¼ 15.27. quinolinylidene-4-yl (compound 15. Compounds 5e8 contained the a-isopropylacetic acid substituent at N3-position of the rhodanine ring. MIC ¼ 15. In an effort to rapidly obtain SAR data. MIC ¼ 15. using a previously reported procedure [35]. The 4-methoxybenzylidene (compound 14. Anti-MRSA activity An in vitro evaluation of in-house rhodanine analogs [31] against MRSA strains resulted in the identification of the moderately potent Scheme 1.6 mg/ mL). a-isobutylacetic acid (compound 11.1.and C5-position of the rhodanine scaffold. We hypothesized that the phenylalanine or other hydrophobic amino acid substituents at N3-position would increase the likelihood of penetration through bacterial cell wall and therefore potentially improving their anti-MRSA activity. we synthesized the next series of rhodanines in the hopes of identifying analogs with greater potency. Rhodanine-3-acetic acid (4) was subjected to Knoevenagel condensation with varied aldehydes to form the target compounds (14e18) as per the procedure reported by us previously [31]. The synthesized compounds 14e18 and 20e22. we obtained compounds 9e13 that had a 2.25 mg/ mL).5828 D. Strains were selected to represent the scope of Staphylococcus variants that might be present in a number of clinical. as well as a series of substituted C5-benzylidenes. Results and discussion 2. The bulky a-n-propylacetic acid (compound 9. The phenylalanine-derived rhodanine intermediate (19) needed for the preparation of target compounds 20e22 was synthesized by first cyclizing the L-phenylalanine with CS2 and sodium a-chloroacetate to form 19. pneumonia patient from Japan.4-dichlorobenzylidene substituent (compound 8. represent nosocomial or hospital-acquired strains from distant global locations and body sites (i. leading to the development of SAR data (Table 1) and identification of a relatively potent analog (compound 12.6 mg/mL) analogs exhibited either comparable or a 2-fold improvement in MIC values.6 mg/mL) and a-benzylacetic acid (compound 12. we decided to purchase structural analogs of compounds 1e3. Collection (ATTC). and BAA-39. MIC ¼ 31. The resulting rhodanine intermediate 19 was subjected to Knoevenagel condensation with varied aldehydes as mentioned above to form the target compounds 20e22.3 for the E isomer [28. U. MIC ¼ 250 mg/mL) analogs exhibited a 4-fold decrease in activity versus the lead compound 1 clearly . Compounds 5e13 were purchased from SigmaeAldrich. We chose rhodanine-3-acetic acid 4 as the scaffold for further SAR exploration.36]. Structures of the anti-MRSA rhodanine hits.2.9 instead of at the calculated chemical shift of 6.4-dichlorobenzylidene substituent at the C5position and various substituents at the N3-position of the rhodanine ring.32e34]. 2. community and quality control situations. along with commercially available rhodanine analogs 5e13. as determined by the chemical shift of the methine proton ranging from 7.A. MIC ¼ 125 mg/mL) exhibited a significant decrease in activity. Following the initial hits with rhodanines 1e3. While 2-chlorobenzylidene (compound 5). vancomycin and penicillin G. were subjected to evaluation against various strains of MRSA (Table 1).e. Chemistry A group of rhodanine analogs (14e18 and 20e22) was synthesized according to Scheme 1.

resulted in the most potent analog of the entire series (compound 22.6e62.9 mg/mL against MRSA ATCC 700787. For example. we hypothesized that these compounds may have the propensity to bind avidly to serum proteins and may influence their anti-MRSA activity.4-dichlorophenyl 2.6 31. MRSA CA ATCC BAA-1680). we determined MIC values of the most potent compounds.4-dichlorophenyl 2. Vanco ¼ vancomycin.5 62.9 1. / European Journal of Medicinal Chemistry 45 (2010) 5827e5832 Table 1 Anti-MRSA activity of compounds 1e3.5 62.6 ND 200 200 250 31. Hardej et al. In order to evaluate our serum protein binding hypothesis.5 62.6 31.6 31. MIC ¼ 3.4-dichlorophenyl 2-chlorophenyl 4-fluorophenyl naphthalene-1-yl 2.8 0.9 7.97 15. Since these structural features (hydrophobic group at C5-position and the acidic function at the N3-position) are present in our rhodanine analogs.25 125 3.6 15.25 62. MIC ¼ 31.6 15.5 250 125 62.9 7. MIC ¼ 3. the application of the molecular hybrid approach to compounds 12 and 17. Subsequently.5 62.D.95 31. 5829 R HOOC S N S O Ar Compda R Ar MRSA ATCC 34404 MRSA ATCC 700787 62.6 125 250 500 250 31.6 31. Ligands binding to site II (also called the indole-benzodiazepine site) on human serum albumin frequently contain aromatic carboxylic acids with a negatively charged acidic group at one end of the molecule located away from a hydrophobic center (e.25 31.25 31. resistance to the fluoroquinolone antibiotics is increasing in CA. Pen-G ¼ penicillin G. they are not .6 15.and 30-fold improvement in activity compared to the respective parent compounds.5 15.4-dichlorophenyl 2.6 125 250 250 250 31.9 62.97 62. The common indirect approach to evaluate the propensity of a molecule to bind serum proteins is the determination of its MIC in the presence of serum.5 3. 8.9 3. nonsteroidal anti-inflammatory drugs [NSAIDs]) [37].5 62. 3 and 17. compounds 5e13 were acquired from SigmaeAldrich whereas compounds 14e22 were synthesized in this report. MRSA ATCC 700698 and MRSA ATCC BAA-39 and MIC ¼ 1. when this approach was utilized for compounds 12 and 3. except ATCC 700787.25 31.5 250 125 ND ND 62. similar to our results (MIC ¼ 0. The molecular hybrid of compounds 12 and 1 resulted in a new analog (compound 20. This finding suggests that these compounds do not significantly bind to serum proteins and this would increase the amount of free drug and thus the anti-MRSA activity.25 15.9 3.5 250 125 62.5 62. Consequently.8 mg/mL for all strains. The MIC value for ciprofloxacin (7.6 MRSA ATCC BAA-39 62.8 0.g.95 mg/mL against MRSA ATCC 34404. we used a molecular hybrid approach. the MIC value for vancomycin is typically 0.25 15. The MIC values obtained for ciprofloxacin.8 0. the MIC values for penicillin G were relatively high (15.95 7..9 mg/mL) with a 4.25 15. MIC ¼ 125 mg/mL) with 8-fold decreased activity compared to 12 and only 2-fold less active compared to 1. The replacement of the 2-nitrobenzylidene moiety in lead compound 1 with 3-phenoxybenzylidene (compound 17.97 31.6 15.25 31.5 250 125 62.4-dichlorophenyl 2.5 3. ND: not determined.6 31.8 0.9 3.25 31.4-dichlorophenyl 2. As expected.97 31.4-dichlorophenyl 3-phenoxyphenyl 62.5 mg/mL) and this is consistent with the fact that MRSA strains are resistant to penicillin G. The results indicated that the MIC values for these compounds were identical in the presence and absence of 10% FBS.6 15. while varying the C5-position substituents present in compounds 1.9 1.6 15.95 7. vancomycin and penicillin G against the various MRSA strains in this study are consistent with previous studies. demonstrating the important role of electron withdrawing groups on C5-benzylidene moiety in influencing the anti-MRSA activity.6 15.and HA-MRSA.25 MRSA CA ATCC BAA-1680 62. thereby potentially decreasing their antimicrobial efficacy.5 mg/mL) is similar to that of earlier report [38].6 125 250 500 250 31. Finally.5e2 mg/mL [21].25 mg/mL) moieties produced a 2-fold increase in activity.4-dichlorophenyl 4-methoxyphenyl quinolin-4-yl styryl 3-phenoxypheny cyclohexyl 2-nitrophenyl 3. 21 and 22 against all MRSA strains in the presence of 10% fetal bovine serum (FBS).6 15.25 MIC mg/mLb MRSA ATCC 700698 62.5 1. which was 62.5 3.25 125 3. it resulted in a new analog (compound 21.5 15. where we chose compound 12 carrying a-benzyl group at N3 substituent. MIC ¼ 31.25 a Compounds 1e3 were synthesized previously [31].5 15. b Results of average values obtained from two independent experiments in duplicates. However.25 62. 5e18 and 20e22.5 1 2 3 5 6 7 8 9 10 11 12 13 14 15 16 17 18 20 21 22 Cipro Vanco Pen-G -H -H -H -i-Pr -i-Pr -i-Pr -i-Pr -n-Pr -methylthioethyl -i-But -benzyl -Me -H -H -H -H -H -benzyl -benzyl -benzyl 2-nitrophenyl 4-nitrophenyl 3.25 62.97 mg/mL).25 15.25 mg/mL) and with cyclohexylidene (compound 18.25 15.6 15.5 15.6 ND 250 250 250 31. However. Cipro ¼ ciprofloxacin.6 15.5 250 125 ND ND 62.5 15. The binding of the molecules to serum proteins can significantly reduce their free concentrations in plasma. the in vitro susceptibility of MRSA strains is highly variable and furthermore.

43 (1H. 700698 and 700787) were purchased from ATCC (Manassas. Calcd. Anal. it is possible that these inhibitors may be blocking functionality of PBP2a. J ¼ 8. 166.2. 8.86 (1H. Alfa Aesar (Ward Hill. The chemical shifts for 1H and 13C are reported in parts per million (d ppm) downfield from tetramethylsilane (TMS) as an internal standard. The 1H NMR data are reported as follows: chemical shift.49. imipenem. 133. d.0. N. 167.. / European Journal of Medicinal Chemistry 45 (2010) 5827e5832 routinely recommended for use in the treatment of MRSA.16 (1H. 4. H. 9.9. MA). Yield ¼ 0. Anal.4 Hz). 148.64 (1H. The anti-MRSA activity of the most potent compounds 8. Experimental 4. Plates used from microdilution techniques were purchased from BectoneDickinson (FranklinLakes. 3. CA) and ciprofloxacin from USP (Rockville.45 g.4.39]. it is possible that the bactericidal compounds in this study could produce their antiMRSA activity via these mechanisms. All compounds were checked for homogeneity by TLC using silica gel as a stationary phase. 7. N. 3. 4. 3. although this remains to be proven experimentally. The antibacterial activity of compounds 21 and 22 against a panel of MRSA strains was significantly greater than that of the reference antibiotics penicillin G and ciprofloxacin. 118. N.4% of calculated values. N.58.6. the mechanism of action of the compounds tested in this study is unknown. 4. d.37 g.12 Hz).95 (1H. OR). CH]). and the residue was dissolved in water. (d) doublet. Typically. s).37 (DCM:MeOH 90:10).73 (2H.45 (4H. 52. 124. d. (m) multiplet. mp >217  C) [41]. N. 129. GA). Therefore. H.34.2 Hz). (Norcross. d).2. and N analyses were performed by Atlantic Microlabs. General procedure for the preparation of 5-benzylidene rhodanine-3-acetic acid derivatives (14e18) Rhodanine-3-acetic acid 4 (0. J ¼ 4. 3/4 H2O: C.3. for C13H11NO4S2: C.8.8. for C14H11NO3S2.64 Hz). 54. Rf ¼ 0. s). and fetal bovine serum from Atlanta Biologicals (Atlanta.47. N. H.5. followed by the addition of ammonium acetate (1. Mueller Hinton II cation-adjusted broth was purchased from Teknova (Hollister.7.70.53. s). 7. Rf ¼ 0.79 (1H.54. 13.8. 1. Currently. d.7. for C14H11NO3S2. 128. 13C NMR (DMSO-d6) d 45. NMR spectra were recorded on a Bruker 400 Avance DPX spectrometer (1H at 400 MHz and 13C at 100 MHz) outfitted with a z-axis gradient probe. 3.37. s). In addition. Found: C.62 (DCM:MeOH 80:20). 7. H.35 (DCM:MeOH 90:10).4. s). compounds 21 and 22 were active against MRSA BAA-39.44 (1H.7. Rf ¼ 0. 4. In addition. 3. (81%).63.5. H. s). 3. gentamicin.N-dimethyl formamide (DMF) was purchased from USB Corporation (Cleveland.1.20 Hz). 4. CH]). 4.15. 167.6. H.20 mmol) and 1. 123. MD). s).7.83. a highly virulent strain. Found: C.73 (1H.8. 193. t. Antibiotics were purchased as follows: vancomycin hydrochloride and penicillin G (potassium salt) from Calbiochem (LaJolla.4. 52. All strains of MRSA (BAA-39.60e3. 13C NMR (DMSO-d6) d 45. compound 22 exhibited only a 2e4-fold higher MIC value than that of vancomycin. OH). 2-(5-(3-Phenoxybenzylidene)-4-oxo-2-thioxothiazolidin-3-yl) acetic acid (17) Brown solid.64 Hz). H. The reaction mixture was refluxed for 1e2 h under inert conditions and the reaction was monitored by TLC.34 (1H.72 (2H. 13.39 g.65. CH] and ArCH]). Thus. 4. Hardej et al. d. m).53. 130. erythromycin. 7.3.7. d. Anal. 166. 7. is characterized by its widespread dissemination and greater likelihood to produce necrotizing pneumonia. J ¼ 8. 3. s. 4. 4. J ¼ 7. 193. Mueller Hinton agar was purchased from HiMedia (India). 21 and 22 remained unchanged in the absence or presence of 10% FBS.84 (3H. This suggests that these compounds may be effective against MRSA strains that are multi-drug resistance. 56. Belgium) and were used as received. although this remains to be proven. J ¼ 4. mp 97e102  C. J ¼ 7. 134. 8.29 (1H. Calcd.18 (1H.3. BAA-1680. (77%). We conclude from this study that the phenylalaninederived compounds 21 and 22 are promising templates for the development of new drugs to treat MRSA infections. 125. 135. The activity of compounds 21 and 22 against the CA-MRSA strain used in this study is important as it is a USA300 genotype.45 (2H.3 g.8. Calcd. 8. 7. 34404. Yield ¼ 0. 7. 8. m). Chemistry-general Melting points (mp) were determined on a Thomas-Hoover capillary melting point apparatus and are uncorrected.3.66 Hz).13 (2H. 1/5H2O: C. It has been reported that several arylalkylidene rhodanines interact with PBPs [27]. 2-(5-(4-Quinolinylidene)-4-oxo-2-thioxothiazolidin-3-yl) acetic acid quinolin-4-yl (15) Yellow solid. and Acros Organics (Antwerp. 4. 21e22) at the N3-position and the electron deficient benzylidene moiety at C5-position of the rhodanine scaffold improves the activity of these compounds against MRSA.57 mmol) was added to a three neck reaction flask equipped with a reflux condenser containing 6 mL anhydrous toluene. 13.57. all MRSA strains harbor the mec gene that codes for the PBP2a protein [12. 162.18]. Rf ¼ 0. (Milwaukee. dried over sodium sulfate and evaporated under vacuum to obtain the purified target compounds 14e18 (compounds 20e22 were purified by column chromatography).4. NJ). The aqueous solution was then acidified with concentrated HCl and the precipitated product was extracted into ethyl acetate.57 mmol of the corresponding aromatic aldehydes.66 (2H. J ¼ 5. 138. mp 220e223  C (lit.44 (2H.5. s). N.39. Yield ¼ 0.10 (2H. 127. 174. 125. antibiotics can produce a bactericidal effect by inhibiting cell wall synthesis or DNA synthesis.9. 7. The reagents for organic synthesis were purchased from Aldrich Chemical Co. 146.4. 124. 1H NMR (DMSO-d6) 4.10 (1H. a strain that has in vitro resistance to clindamycin. multiplicity (s) singlet.42 g. 1H NMR (DMSO-d6) d 4.61. 134.1. leucine (compound 11) and phenylalanine (compounds 12. 166. tetracycline.9.6. 120. 115. Inc. 54. WI). invasive skin and skin structure infections and sepsis [2.44. 13C NMR (DMSO-d6) d 47. The reaction mixture was evaporated under vacuum. Conclusions We report the synthesis and anti-MRSA activity of a series of glycine and phenylalanine-derived rhodanine analogs. 50. 4. TCI America (Portland. 150. (87%). 130. Most importantly.5.53. 50.5830 D. 7. (84%). GA) and the observed values were within Æ0. . The USA300. mp 178e180  C [31].68 Hz).51 (1H. mp 210e214  C (lit.34 (DCM:MeOH 94:6). CA). This could be due to these substituents increasing their ability to penetrate the bacterial cell wall. The C. tobramycin. amoxicillin and five different cephalosporins.6. 7.9. 2-(5-(4-Methoxybenzylidene)-4-oxo-2-thioxothiazolidin-3-yl) acetic acid (14) Yellow solid. The SAR study clearly suggests the presence of the hydrophobic side chains of valine (compound 8). t. 128. 1H NMR (DMSOd6) d 3. 167.6. mp 190e193  C) [40]. 7.1. 2-(5-(Cinnamylidene)-4-oxo-2-thioxothiazolidin-3-yl)acetic acid styryl (16) Yellow solid. (t) triplet.2.9. 7.74 (2H. VA). 1H NMR (DMSO-d6) d 4. J ¼ 4. d. Compounds 21 and 22 did produce a bactericidal action on selected MRSA strains. Found: C.6.7.4. s. 193. Yield ¼ 0. d.

JAMA 290 (2003) 2976e2984. Boxrud. 144. 124. One hundred microliters of cell suspension was added to each well. Epidemiology of methicillin-resistant staphylococcus aureus. Med. 46 (2008) S344eS349.8. Morrison. 65. Dis. 45. Klevens. 116. L. 1/2H2O: C. m). 134.6. CH]). 2.7. 2-(5-(2-Nitrobenzylidene)-4-oxo-2-thioxothiazolidin-3-yl)-3phenylpropanoic acid (20) Yellow solid.-Mon. Corey. 7. N. m).51.22 (1H. CH]).3. J. 50. 128. Yield ¼ 0. 3. 7. s. 7. M. 1H NMR (DMSO-d6) d 1. t.22 (2H. J ¼ 7.2. 7. K. After 24 h. 58. 194. m).3. 122. McLaughlin from the Department of Chemistry. Lynfield. G. 7. Craig. 13C NMR (DMSO-d6) d 33.E.3. 57. 1H NMR (DMSO-d6) d 3. 320 (1989) 1188e1196.90 (1H. 3. Borchardt.46 (1H. Boucher. Johnson. 128.6 (DCM:MeOH 95:5).1. s). 167. Etienne.10. dried over anhydrous sodium sulfate and evaporated under vacuum and the residue was purified by column chromatography to afford a brown liquid. H. 124.0. d. Rf ¼ 0. 3. m). s). 13C NMR (DMSO-d6) d 25.J ¼ 7. 120. 168. R. 120. Hamilton-Miller. Calcd.4. s).37 (1H.49 (2H.4.8. O’Boyle. 129. Drugs 69 (2009) 693e716. We thank Dr. 30.21 (5H. 5. 56. J ¼ 6. McDougal.78. Yield ¼ 0. 5. N.98 Hz). 58. 148.S. 3.37 (1H. Gershman. Then. 130. 8. Each compound was tested against all strains of MRSA minimally in triplicate. 119. Florida for his help in interpretation of the spectral data.48. H. 167.72. N. H. 123. [3] M.N. 4.44.2. 2-(5-(Cyclohexylidene)-4-oxo-2-thioxothiazolidin-3-yl)acetic acid (18) (lit.4. 4. N. (30%). The reaction mixture was neutralized with saturated NaHCO3 solution. Methicillin-resistant staphylococcus aureus.17 (3H.A. d.0. Hardej et al. 2.82). m).34.51 g. 3. Danila.4.27 (2H. 7. 4. R. Rf ¼ 0. 30.6. 43. 119. 52. for C25H19NO4S2. Brumfitt. Rf ¼ 0.9. J ¼ 9. 13.8. [4] T.6. J ¼ 7. 50.8. For the active bacterial core surveillance (ABCs) MRSA investigators.5% of the total media volume. 166. Petit.6. N. mp Brown solid.8.8.7. LeDell.F.4. Anal. 130. L. Found: C. H. 126. 156. 3. 131.6.2.0.3 mmol) was dissolved with sodium hydroxide (1. N. 7.M. 4.08 Hz). The MIC assays were carried out in 96 well sterile plates. 65. 121. J ¼ 8.88 Hz).0. Found: C. s). 13. E.0. 135. . Como-Sabetti. 132.55 (1H. Carey.89 (DCM:MeOH 80:20). J ¼ 7. 116.7. Initially.M. J. d.69 (1H. for C19H14N2O5S2. 130. C 5. 121. H. Dumyati.83.3. Clin. 129. Calcd. 41.6.4. Harrison. 7.99. 5.51 (2H.6.0. John’sUniversity are gratefully acknowledged. H.5. 7.23 (1H.K.1. 193. N. Test for antibacterial activity All purchased and synthesized compounds were tested for their in vitro antibacterial activity against all strains of MRSA by performing a microdilution minimal inhibitory concentration (MIC) technique. 7. Lee Moffitt Cancer Center & Research Institute.12. 7.86 (1H. 131. [2] R.79. Gordon. A. Then the reaction mixture was acidified with dilute HCl until pH 1.87 (1H.T.48 (2H.1.7 (DCM:MeOH 95:05).68 (DCM:MeOH 95:5). 130. 13. 134. N.B.2 Hz). Appropriate cell concentrations of each strain of MRSA were made in Mueller Hinton broth by adjusting the turbidity to a 0. Decker. m).59 (1H. 128. Med. The resultant solution was acidified again with dilute HCl. s. 31.3.94 (1H.6. Found: C. R. 133. s).1. 129.087 g. J. t.3. Ray. 4. (25%).N-dimethyl formamide (DMF) and then adding cation-adjusted Mueller Hinton broth. J. m).7.20. The plates were incubated at 37  C overnight. s). Preparation of 2-(4-oxo-2-thioxothiazolidin-3-yl)-3phenylpropanoic acid (19) In a round-bottomed flask equipped with a magnetic stirrer.9.S. 131.30.3 mmol) was added and stirring was continued at 23  C for 3 h. 7. Engl.22 g. All the experiments were performed in triplicate.3.D. 7.17. J ¼ 5. / European Journal of Medicinal Chemistry 45 (2010) 5827e5832 5831 J ¼ 8. Chavez. Calcd. 339 (1998) 520e532. 137. 2 CH2Cl2: C. Found: C.and health care-associated methicillin-resistant staphylococcus aureus infection. 127. Yield ¼ 0.6. 157. An aqueous solution of sodium chloroacetate (3.94 (1H. 132. G. Mark L. Vandenesch.K. S.8. Fosheim.28 (1H. 46 (2008) S350eS359.09. mp 63e65  C.71. carbon disulfide (2. 4. s). a 2 mg/mL concentration of a compound was made by dissolving it in N. 125. 192. Health care-associated MRSA versus communityassociated MRSA.A. Lowy.3 mmol) was added to the reaction mixture.79 (1H.: C. 7.2. d. 131. Acknowledgements The financial support and resources provided by the College of Pharmacy of St.8.42 (1H.2. Dis. F. m). C.68. 129.3. 115.5. 4.93.5. 13. S.33. 1H NMR (DMSO-d6) d 3.7. d. 5 mL was drawn from the wells and spotted onto suitable agar plates.2. [7] T.6.75.7.5. Found: C. 25.51.9. J ¼ 7. 13C NMR (DMSO-d6) d 33.1. phenylalanine (5 g. Comparison of community.1% survival in the subculture. 33.09.8.6.3.5. mp 138e140  C. Calcd.11. 13. 133.0. 136. Townes. 30. 140. [6] F. J ¼ 7.6 Hz). H. 7.0. D. N. t. 7. 30. Serial dilution of the compounds was made in Mueller Hinton broth (100 mL).52.80. J. 2-(5-(3-Phenoxybenzylidene)-4-oxo-2-thioxothiazolidin-3yl)-3-phenylpropanoic acid (22) Yellow solid. S. 1H NMR (DMSO-d6) d 3.7. t. F. mp 138e141 209e211  C) [36].79. 127.79 (2H.0.91. m).2.0. J ¼ 4. H. 165.2.2.1. Yield ¼ 2. Anal. for C19H13Cl2NO3S2. 4. [5] W.14 (1H. 3.4-Dichlorobenzylidene)-4-oxo-2-thioxothiazolidin-3yl)-3-phenylpropanoic acid (21) Yellow solid.98.54 (1H. Staphylococcus aureus infections.M. Clin. References [1] R. 4. Lowy. (86%). 13C NMR (DMSO-d6) d 48. University of South Florida and the Department of Drug Discovery. The quantity of DMF added never exceeded 0. 132. Anal.68 Hz).58 (1H.39 g.08 (2H. s. 120.44 (2H. m). S. 132. Infect.3. 2-(5-(3. s).52 (1H.10.18 (5H. d. 192. s. 193. 54 (2008) 763e768.21 g. 2. The MIC of each compound was recorded as the well showing no turbidity when compared to control wells. 135.41 g. The MBC read 18 h later was defined as the lowest concentration of compound or standard reference antibiotics that resulted in 0. 51. 1. d. 3. J ¼ 7. 1/5C6H14: C.84 Hz).9. Control wells contained Mueller Hinton broth and cell suspension but no compound. 125. K.5. 126.04 (1H. 7.54 (DCM:MeOH 95:5). 8. S.0 and refluxed overnight.66 (6H. 7.8.45 (3H. J ¼ 7. t.65 (1H. 43.40 (1H. 155.J. 4.41 (2H. JAMA 298 (2007) 1763e1771. R.3. 5. 4. 6. s).75 (1H.2.3. m). J ¼ 8.81. H. invasive methicillin-resistant staphylococcus aureus infections in the United States. s). 7. [8] H.H. 5. 4.89. Patel. 4.K. 124. N.55 g.9. 72. d.3 mmol) in water (25 mL). 167. t. Anal. Infect. Community-associated methicillin-resistant staphylococcus aureus infections: epidemiology. H.4 Hz).0.H. Pathogenesis of methicillin-resistant staphylococcus aureus infection.1. J ¼ 7. Yield ¼ 0. s).6. S.93. for C12H15NO3S2: C. 125. Lynfield. MBCs were established by extending the MIC procedure to the evaluation of bactericidal activity.18 (8H. Nadle.0. Naimi.85 (1H.5 McFarlane Standard and then adding 0. Anal. Fridkin. Calcd. 13.1. Engl. Rf ¼ 0. 124. Tampa. 166. H.32 Hz).3 g. Zell. for C18H13NO4S2. 169. 13C NMR (DMSO-d6) d 29. 133. N. 123. m).4.66 (2H.52 Hz).10. (44%).0.52 Hz).0. 30. 3. 7. G.06.2 mL of the suspension to 40 mL of fresh Mueller Hinton Broth. 166.5. 1H NMR (DMSO-d6) d 3. recognition and management. J. 117. C.R. CH]). CH]).68 Hz).44 Hz). t. mp 170e175  C. 7. The cyclized product was extracted in ethyl acetate. 7. CH]).28 (5H.2. N. 158. 4. Rf ¼ 0. s).78 Hz).D. K. (89%). Fridkin. Dis. which was stirred vigorously overnight. 169.87 (1H. 59.

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