Abstract
Cyclohex-3-enyl(5-phenyl-4H-1,2,4-triazol-3-yl)methanol (MSDRT 12) is a novel triazole-based antitubercular compound with two chiral centres. Evaluation of the enantio-specific antitubercular activity has established that the stereoisomer 3 of MSDRT12 (Isomer 3) was the most potent isomer with a minimum inhibitory concentration of 0.78 μg/mL. The other stereoisomers show negligible or no activity. A sensitive, simple, specific, precise and accurate chiral chromatographic method for the direct analysis of the four stereoisomers of MSDRT 12 and the active Isomer 3 has been developed and validated. The method has also been validated for analysing the stereoisomeric impurities Isomer 1, Isomer 2 and Isomer 4 in the active Isomer 3. The separation of the four stereoisomers of MSDRT 12 was achieved using an immobilized polysaccharide-based column, Chiralpak ID with amylose tris(3-chlorophenylcarbamate) as the chiral selector. The separation was performed using a mixture of n-hexane, isopropyl alcohol, ethanol and diethylamine (60:35:5:0.1 v/v/v/v) at a flow rate of 1 mL/min. The method offers excellent separation of the four stereoisomers with resolution more than 1.5 and tailing factor <1.5. The standard curves were linear over the concentration range 5–500 μg/mL and 0.40–505 μg/mL for MSDRT 12 and Isomer 3, respectively. Excellent linearity in the range 0.4–5 μg/mL was obtained for Isomer 1, Isomer 2 and Isomer 4 and these stereoisomeric impurities could be accurately and precisely quantified at a level of 0.1 % of the active isomer.
References
Yingru Z, Dauh RW, David BW, Adrienne AT (2005) Enantioselective chromatography in drug discovery. DDT 10(8):571–577
Margareta EA, David A, Adrian C, Johan R, Gunnar H (2003) Evaluation of generic chiral liquid chromatography screens for pharmaceutical analysis. J. Chromatogr. A 1005(1–2):83–101
Christopher JW, Peter S, Mirlinda B, Gouker Joe, Fairchild Jacob (2006) Comparison of multi-parallel microfluidic HPLC instruments for high throughput analyses in support of pharmaceutical process. J. Liq. Chromatogr. Relat. Tech. 29:2185–2200
Ilkay K, Guniz K, Sevim R, Muammer K (2001) Some 3-thioxo/Alkylthio-1,2,4-triazoles with a substituted thiourea moiety as possible anti-mycobacterials. Bioorg. Med. Chem. Lett. 11:1703–1707
Lenka Z, Vera K, Jan K, Karel W, Jarmila K (2004) Synthesis and antimycobacterial activity of pyridylmethylsulfanyl and naphthylmethylsulfanyl derivatives of benzazoles, 1,2,4-Triazole, and pyridine-2-carbothioamide/-2-carbonitrile. Arch. Pharm. Pharm. Med. Chem. 337:549–555
Alireza F, Zahra K, Fatemeh S (2003) Antituberculosis agents VIII––synthesis and in vitro antimycobacterial activity of alkyl α-[5-(5-nitro-2-thienyl)-1,3,4-thiadiazole-2-ylthio] acetates. Farmaco 58(11):1073–1076
Nutan HP, Ranjeet B, Manoj K, Nilesh T, Ray M, Rajan M, Mariam D (2013) Novel 4H-1,2,4-triazol-3-yl cycloalkanols as potent antitubercular agents. Med. Chem. Res. 22:401–408
Shekar R, Sinha BN, Mukhopadhya A, Degani MS (2013) Isolation and evaluation of enantiospecific antitubercular activity of a novel triazole compound, doi:10.3797/scipharm.1308-15
Daniel WA, Bo Z (2001) Chiral stationary phases for HPLC. Anal. Chem. 73(19):557A–561A
Yoshio O, Eiji Y (1998) Polysaccharide derivatives for chromatographic separation of enantiomers. Chem. Int. Ed. 37:1020–1043
Yoshio O, Mitsunobu K, Koichi H (1984) Useful chiral packing materials for high performance liquid chromatographic resolution of enantiomers: phenylcarbamates of polysaccharides coated on silica gel. J. Am. Chem. Soc. 106:5357–5359
International Conference on Harmonization (2005) ICH guidelines on validation of analytical procedures: text and methodology Q2 (R1)
International Conference on Harmonization (2002) ICH guidelines on analytical method validation. In: Proceedings of the International Convention on Quality for the Pharmaceutical Industry, Toronto
Acknowledgments
The authors would like to thank the management of Lotus Labs Pvt. Ltd. for supporting this work. Technical assistance from Mr. Lakshminarayana, Ms. Vennila R and Mr. Ramkumar Dubey is greatly acknowledged. The authors would also like to thank Head, Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, India for constant encouragement and support for this work.
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Shekar, R., Sinha, B.N., Mukhopadhya, A. et al. Chiral HPLC Method for the Novel Triazole Antitubercular Compound MSDRT 12. Chromatographia 77, 511–516 (2014). https://doi.org/10.1007/s10337-013-2613-5
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DOI: https://doi.org/10.1007/s10337-013-2613-5