Obtaining and Certifying Primary Reference Standard of 6,8-Dimethyl-2-Piperidinomethyl-2,3-Dihydrothiazolo[2,3-F]xanthine

INTRODUCTION. An effective quality control system for medicinal products is impossible without reference standards used for validation and verification of analytical procedures. It is especially relevant to develop reference standards for quality control of new active pharmaceutical ingredients used to develop medicinal products.

AIM. This study aimed to develop a method for obtaining and certifying a primary reference standard of 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F] xanthine to be used in quality control of medicinal products.

MATERIALS AND METHODS. A reference standard was obtained by recrystallising 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine from ethanol. The structure of the reference standard was defined by nuclear magnetic resonance and infrared spectroscopy; purity was measured by mass balance, high-performance liquid chromatography (related impurities), and titration (non-aqueous acidimetry).

RESULTS. The study assessed physical and chemical properties of a reference standard for 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine. This included structure elucidation by infrared and nuclear magnetic resonance spectroscopy; loss on drying 0.073±0.015%; sulfate ash 0.080±0.009%; related substances (not found); heavy metal impurities (not more than 0.002%); elemental composition (C — 53.68±0.17%; H — 6.32±0.02%; N — 20.81±0.09%; O — 9.52±0.06%; S — 9.57±0.04%); quantitative determination by non-aqueous titration 99.74±0.12%, and mass balance 99.85±0.01%.

INTRODUCTION. An effective quality control system for medicinal products is impossible without reference standards used for validation and verification of analytical procedures. It is especially relevant to develop reference standards for quality control of new active pharmaceutical ingredients used to develop medicinal products.

AIM. This study aimed to develop a method for obtaining and certifying a primary reference standard of 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F] xanthine to be used in quality control of medicinal products.

MATERIALS AND METHODS. A reference standard was obtained by recrystallising 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine from ethanol. The structure of the reference standard was defined by nuclear magnetic resonance and infrared spectroscopy; purity was measured by mass balance, high-performance liquid chromatography (related impurities), and titration (non-aqueous acidimetry).

RESULTS. The study assessed physical and chemical properties of a reference standard for 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine. This included structure elucidation by infrared and nuclear magnetic resonance spectroscopy; loss on drying 0.073±0.015%; sulfate ash 0.080±0.009%; related substances (not found); heavy metal impurities (not more than 0.002%); elemental composition (C — 53.68±0.17%; H — 6.32±0.02%; N — 20.81±0.09%; O — 9.52±0.06%; S — 9.57±0.04%); quantitative determination by non-aqueous titration 99.74±0.12%, and mass balance 99.85±0.01%.

INTRODUCTION. An effective quality control system for medicinal products is impossible without reference standards used for validation and verification of analytical procedures. It is especially relevant to develop reference standards for quality control of new active pharmaceutical ingredients used to develop medicinal products.

AIM. This study aimed to develop a method for obtaining and certifying a primary reference standard of 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F] xanthine to be used in quality control of medicinal products.

MATERIALS AND METHODS. A reference standard was obtained by recrystallising 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine from ethanol. The structure of the reference standard was defined by nuclear magnetic resonance and infrared spectroscopy; purity was measured by mass balance, high-performance liquid chromatography (related impurities), and titration (non-aqueous acidimetry).

RESULTS. The study assessed physical and chemical properties of a reference standard for 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine. This included structure elucidation by infrared and nuclear magnetic resonance spectroscopy; loss on drying 0.073±0.015%; sulfate ash 0.080±0.009%; related substances (not found); heavy metal impurities (not more than 0.002%); elemental composition (C — 53.68±0.17%; H — 6.32±0.02%; N — 20.81±0.09%; O — 9.52±0.06%; S — 9.57±0.04%); quantitative determination by non-aqueous titration 99.74±0.12%, and mass balance 99.85±0.01%.

CONCLUSION. We have developed production process of 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine reference standard. Physical and chemical properties of the above standard comply with the requirements and allow us to recommend 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine as a reference in the quality control of medicinal products.

1. Devarbhavi H, Asrani SK, Arab JP, et al. Global burden of liver disease: 2023 update. J Hepatol. 2023;79(2):516–37. https://doi.org/10.1016/j.jhep.2023.03.017

2. Neshat SY, Quiroz VM, Wang Y, et al. Liver disease: Induction, progression, immunological mechanisms, and therapeutic interventions. Int J Mol Sci. 2021;22(13):6777. https://doi.org/10.3390/ijms22136777

3. Singh N, Shreshtha AK, Thakur MS, Patra S. Xanthine scaffold: scope and potential in drug development. Heliyon. 2018;4(10):e00829. https://doi.org/10.1016/j.heliyon.2018.e00829

4. Specker E, Wesolowski R, Schütz A, et al. Structure-based design of xanthine-imidazopyridines and ‑imidazothiazoles as highly potent and in vivo efficacious tryptophan hydroxylase inhibitors. J Med Chem. 2023;66(21):14866–96. https://doi.org/10.1021/acs.jmedchem.3c01454

5. Specker E, Matthes S, Wesolowski R, et al. Structure-based design of xanthine-benzimidazole derivatives as novel and potent tryptophan hydroxylase inhibitors. J Med Chem. 2022;65(16):11126–49. https://doi.org/10.1021/acs.jmedchem.2c00598

6. Airhihen B, Pavanello L, Jadhav GP, et al. 1-Hydroxy--xanthine derivatives inhibit the human Caf1 nuclease and Caf1-containing nuclease complexes via Mg²⁺-dependent binding. FEBS Open Bio. 2019;9(4):717–27. https://doi.org/10.1002/2211-5463.12605

7. Saratikov AC, Novozheeva TP, Ahmedzhanov RR. Efficacy of enzyme-inducing agents in rats with intrahepatic cholestasis. Bulletin of Experimental Biology and Medicine. 2002;134(3):271–3. https://doi.org/10.1023/A:1021563703378

8. Nikitina IL, Bagmanova IV, Alekhin EK, Haliullin FA. Isoenzyme profile of the new P450 inducer dihydrothiazoloxanthine hepazan. Gastro-Bulletin. 2001;(2–3):59 (In Russ.).

9. Samorodov AV, Kamilov FKh, Khalimov AR, et al. Antiaggregatory activity of salts of 7-thietanilxanthine derivatives under conditions in vitro. Modern Problems of Science and Education. 2016;(5):32 (In Russ.). EDN: WWVFMR

10. Khaliullin FA, Shabalina YV, Davlyatova GG, Valeeva LA. Synthesis and antidepressant properties of 3-methyl-7-(1,1-dioxothietan-3-YL)-8-cyclohexylamino-1-ethyl-1H-purine-2,6(3H,7H)-dione. Pharm Chem J. 2018;51:1049–52. https://doi.org/10.1007/s11094-018-1738-0

11. Belousov MV, Yusubov MS, Khaliullin FA, et al. Method for preparing 1,3-dimethyl-7-(piperidin-1-ylmethyl)-6,7-dihydrothiazolo[ 3,2-F]purin-2,4(1H,3H)-dione. Patent of the Russian Federation No. 2660653; 2018 (In Russ.). EDN: ZEIQKD

12. Gazizova IR, Alekhin EK. Antitoxic properties of the new inducer of the microsomal enzyme system hepazan. Experimental and Clinical Pharmacology. 2001;(5):96–8 (In Russ.). EDN: MPITJB

13. Nikitin NA, Khaliullin FA, Alekhin EK, et al. Quantitative structure — Activity relationships for microsomal enzymatic system modulants. Part II: Inductors. Pharm Chem J. 2001;35:328–32. EDN: VMPGRN. https://doi.org/10.1023/A:1012397722001

14. Merkulov VA, Sakanyan EI, Klimov VI, et al. Modern approaches to the development of reference standards for quality assessment of drug substances. Pharm Chem J. 2016; 49:776–8. https://doi.org/10.1007/s11094-016-1369-2

15. Moiseev SV, Kuz’mina NE, Luttseva AI. NMR as used in the Russian and foreign pharmacopoeias for quality control of medicinal products. Bulletin of the Scientific Centre for Expert Evaluation of Medicinal Products. Regulatory Research and Medicine Evaluation. 2022;12(1):8–23 (In Russ.). https://doi.org/10.30895/1991-2919-2022-12-1-8-23

16. Kuzmina NE, Moiseev SV, Yashkir VA, Osintseva EV. The possibility of the nuclear magnetic resonance methods using for reference standards certification. Measurement Standards. Reference Materials. 2014;(2):19–25 (In Russ.). EDN: SIWAVZ

17. Krylov VI, Yashkir VA, Braun AV, et al. Development of methods for the synthesis of malathion-D6, chlorophos-D6, and dichlorophos-D6 for use as internal standards in the analysis of medicinal plant raw materials and herbal medicinal products. Bulletin of the Scientific Centre for Expert Evaluation of Medicinal Products. Regulatory Research and Medicine Evaluation. 2023;13(3):411–8 (In Russ.). https://doi.org/10.30895/1991-2919-2023-517

18. Epshtein NA. Certification of reference samples in pharmacy: Mass balance method. Pharm Chem J. 2018;51:928–37. https://doi.org/10.1007/s11094-018-1718-4

19. Khaliullin FA, Alekhin EK, Krasilova IL, et al. Method of preparing 6,8-dimethyl-2-piperidinomethyl-2,3-dihydrothiazolo[2,3-F]xanthine. Patent of the Russian Federation No. 2161160; 1999. EDN: TZFMCI

20. Urazbaev MA, Khaliullin FA, Samorodov AV, Kamilov FKh. Synthesis of salts 2-[1-benzyl-3-methyl-7-(thietan-3-yl) xanthinyl-8-thio] acetic acid and their impact on the hemostasis system. Journal of Scientific Articles «Health and Education Millennium». 2017;19(10):374–8 (In Russ.). EDN: ZATPLT