Obtaining the Enoxaparin Sodium Substance Equivalent to the Original Clexane® and Lovenox®. The Selection of Technological Parameters and Optimization of the “Greenness” of the Purification Stage

Authors

DOI:

https://doi.org/10.24959/ophcj.23.290670

Keywords:

enoxaparin sodium, low-molecular-weight heparin, technological parameters, compositional analysis, HSQC, size-exclusion chromatography, green chemistry, E-factor, solvent regeneration

Abstract

The aim of the study was to adjust and optimize the purification stage of crude enoxaparin sodium to obtain a substance equivalent to the original drugs Clexane® and Lovenox® according to the criteria specified by the FDA. The purification stage involves the reprecipitation of crude enoxaparin in methanol. Determining the ratio of solvents required for the reprecipitation is important for studying the correlation between the experimental conditions of the technological process and the structural characteristics of enoxaparin samples. In the study, the method of purification of enoxaparin sodium described in the patent was assessed, and the following variations of the MeOH:H2O solvent ratio were selected – 4:1; 2:1; 1:1. The obtained samples of enoxaparin sodium were analyzed according to the in-house specification developed on the basis of the pharmacopoeial monograph, as well as by non-pharmacopoeial methods, such as two-dimensional NMR spectroscopy (HSQC) and size exclusion chromatography (SEC) for detailed characterization of the molecule. Strategies of greening of the enoxaparin sodium purification stage by reducing the E-factor were also considered in the study. Considering the principles of “green” chemistry, the method of purification of crude enoxaparin sodium was optimized by the solvent regeneration. It was experimentally possible to demonstrate the effect of the solvent ratio at the stage of purification of crude enoxaparin on the composition, as well as on the number and distribution of oligosaccharide fractions in the molecule. Based on the results of the study, it can be concluded that the ratio of MeOH:H2O=1:1 allows obtaining samples that are closest to Clexane® and Lovenox® in terms of the molecular weight distribution profile and the composition profile. The E-factor was also reduced from 14 to 5.25 by solvent regeneration.

Supporting Agency

  • The research was carried out with the financial support of JSC Farmak (Kyiv, Ukraine).

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References

  1. Baytas, S. N.; Linhardt, R. J. Advances in the Preparation and Synthesis of Heparin and Related Products. Drug Discovery Today 2020, 25 (12), 2095 - 2109. https://doi.org/10.1016/j.drudis.2020.09.011.
    |
  2. Lazrak, H. H.; René, É.; Elftouh, N.; Leblanc, M.; Lafrance, J.-P. Safety of Low-Molecular-Weight Heparin Compared to Unfractionated Heparin in Hemodialysis: A Systematic Review and Meta-Analysis. BMC Nephrology 2017, 18 (1), 187. https://doi.org/10.1186/s12882-017-0596-4.
    |
  3. Green, M. S.; Tellor, K. B.; Buckallew, A. R. Safety and Efficacy of Enoxaparin Compared with Unfractionated Heparin for Venous Thromboembolism Prophylaxis in Hemodialysis Patients. Hospital Pharmacy 2017, 52 (9), 623 - 627. https://doi.org/10.1177/0018578717724799.
    |
  4. Iqbal, Z.; Sadaf, S. Scientific Considerations in the Regulatory Approval of Generic (or Biosimilar) Version of Enoxaparin Sodium – a Lifesaving Carbohydrate Polymer. Regulatory Toxicology and Pharmacology 2023, 143, 105446 - 105446. https://doi.org/10.1016/j.yrtph.2023.105446.
    |
  5. Xie, S.; Guan, Y.; Zhu, P.; Li, F.; Yu, M.; Linhardt, R. J.; Chi, L.; Jin, L. Preparation of Low Molecular Weight Heparins from Bovine and Ovine Heparins Using Nitrous Acid Degradation. Carbohydrate Polymers 2018, 197, 83 - 91. https://doi.org/10.1016/j.carbpol.2018.05.070.
    |
  6. Fu, L.; Suflita, M.; Linhardt, R. J. Bioengineered Heparins and Heparan Sulfates. Advanced Drug Delivery Reviews 2016, 97, 237 - 249. https://doi.org/10.1016/j.addr.2015.11.002.
    |
  7. Imberti, D.; Marietta, M.; Polo Friz, H.; Cimminiello, C. The Introduction of Biosimilars of Low Molecular Weight Heparins in Europe: A Critical Review and Reappraisal Endorsed by the Italian Society for Haemostasis and Thrombosis (SISET) and the Italian Society for Angiology and Vascular Medicine (SIAPAV). Thrombosis Journal 2017, 15 (1), 13. https://doi.org/10.1186/s12959-017-0136-2.
    |
  8. Harenberg, J.; Walenga, J.; Torri, G.; Dahl, O. E.; Drouet, L.; Fareed, J. Update of the Recommendations on Biosimilar Low-Molecular-Weight Heparins from the Scientific Subcommittee on Control of Anticoagulation of the International Society on Thrombosis and Haemostasis. Journal of Thrombosis and Haemostasis 2013, 11 (7), 1421 - 1425. https://doi.org/10.1111/jth.12269.
    |
  9. Guerrini, M.; Rudd, T. R.; Mauri, L.; Macchi, E.; Fareed, J.; Yates, E. A.; Naggi, A.; Torri, G. Differentiation of Generic Enoxaparins Marketed in the United States by Employing NMR and Multivariate Analysis. Analytical Chemistry 2015, 87 (16), 8275 - 8283. https://doi.org/10.1021/acs.analchem.5b01366.
    |
  10. Harenberg, J.; Cimminiello, C.; Agnelli, G.; Di Minno, G.; Polo Friz, H.; Prandoni, P.; Scaglione, F. Biosimilars of low-molecular-weight heparin products: fostering competition or reducing ‘biodiversity’? Journal of Thrombosis and Haemostasis 2016, 14 (3), 421 - https://doi.org/10.1111/jth.13237.
    |
  11. Bovsunovska, Y.; Rudiuk, V.; Mishchenko, V.; Georgiyants, V. Obtaining the substance enoxaparin sodium equivalent to the original Clexane® and Lovenox®. Selection of technological parameters of the key stage of the synthesis. ScienceRise: Pharmaceutical Science 2023, 2, 46 - https://doi.org/10.15587/2519-4852.2023.277735.
  12. Debrie, R. Mixtures of particular LMW heparinic polysaccharides for the prophylaxis/treatment of acute thrombotic events. US Patent US5389618A, Feb 14, 1995.
  13. Liu, X.; St Ange, K.; Wang, X.; Lin, L.; Zhang, F.; Chi, L.; Linhardt, R. J. Parent Heparin and Daughter LMW Heparin Correlation Analysis Using LC-MS and NMR. Analytica Chimica Acta 2017, 961, 91 - 99. https://doi.org/10.1016/j.aca.2017.01.042.
    |
  14. Beecher, C. N.; Manighalam, M. S.; Nwachuku, A. F.; Larive, C. K. Screening Enoxaparin Tetrasaccharide SEC Fractions for 3-O-Sulfo-N-Sulfoglucosamine Residues Using [1H,15N] HSQC NMR. Analytical and Bioanalytical Chemistry 2016, 408 (6), 1545 - 1555. https://doi.org/10.1007/s00216-015-9231-z.
    |
  15. Ingle, R. G.; Agarwal, A. A World of Low Molecular Weight Heparins (LMWHs) Enoxaparin as a Promising Moiety – A Review. Carbohydrate Polymers 2014, 106, 148 - 153. https://doi.org/10.1016/j.carbpol.2014.01.100.
    |
  16. Sheldon, R. A. Metrics of Green Chemistry and Sustainability: Past, Present, and Future. ACS Sustainable Chemistry & Engineering 2018, 6 (1), 32 - https://doi.org/10.1021/acssuschemeng.7b03505.
  17. Sheldon, R. A. The E Factor 25 Years on: The Rise of Green Chemistry and Sustainability. Green Chemistry 2017, 19 (1), 18 - 43. https://doi.org/10.1039/c6gc02157c.
  18. Vedula, M. S.; Kosgi, S.; Mantena, N. D.; Aryasomayajula, R. Process for the preparation of low molecular weight heparin. WO2019116217A2, Jun 20, 2019.
  19. Prat, D.; Hayler, J.; Wells, A. S. A Survey of Solvent Selection Guides. Green Chemistry 2014, 16 (10), 4546 - 4551. https://doi.org/10.1039/c4gc01149j.
  20. Sheldon, R. A.; Bode, M. L.; Akakios, S. G. Metrics of green chemistry: Waste minimization. Current Opinion in Green and Sustainable Chemistry 2022, 33, 100569. https://doi.org/10.1016/j.cogsc.2021.100569.

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Published

2023-11-07

How to Cite

(1)
Bovsunovska, Y. V.; Rudiuk, V. V.; Harna, N. V.; Holovchenko, O. S.; Georgiyants, V. A. Obtaining the Enoxaparin Sodium Substance Equivalent to the Original Clexane® and Lovenox®. The Selection of Technological Parameters and Optimization of the “Greenness” of the Purification Stage. J. Org. Pharm. Chem. 2023, 21, 38-49.

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Original Researches