Acta Scientific Biotechnology

Research Article Volume 1 Issue 11

Finding the Biological Properties from a Library of Steroids

Dhvani H Kuntawala1, Glen JP McCann2* and Neill J Horley3*

1Department of Life Sciences, De Montfort University, United Kingdom
2Department of Life Sciences, De Montfort University, United Kingdom
3Department of Life Sciences, De Montfort University, United Kingdom

*Corresponding Author: Glen JP McCann, De Montfort University and Neill J Horley, De Montfort University.

Received: September 26, 2020; Published: October 21, 2020

×

Abstract

  Most pharmaceutical companies have access to many untested libraries of compounds. Virtual in silico testing of these large libraries are limited to the quality of known data and even then, would fail to discover new classes of compounds. Therefore, testing in vitro in the laboratory is still very important. However, the cost of screening large libraries for existing and new biological target or properties may not be cost effective without initial leads. The aim of this study was to evaluate 90 steroidal compounds, utilizing various rapid and relatively cheap spectrophotometric testing methods. The assays used were to detect antioxidant, antifungal and inhibition activity of CYP3A4 by using 96-well microplate methods. The rationale for choosing these assays was that steroids are known to possess such activities and that the library of steroids used have quite variable structures. This may find new or enhanced potencies for these targets. The methods used was the ABTS and DPPH antioxidant assays, antimicrobial zone of inhibition testing on agar, and inhibition screening using CYP3A4 with the fluorescent substrate DBF. The main observations from the study were that several of the compounds possessed relative activities. Antioxidant assays both using ABTS and DPPH found 2 - 3 compounds with parity or better antioxidant activity (compounds 51, 77 and 81) than the control Trolox. Having a few exceptions, most of the test compounds had some scavenging antioxidant activity. For antifungal activity via yeast growth inhibition (compounds 80- and 86-mm zones of inhibition). CYP3A4 inhibition IC50 values were (56.20, 710.9, 337536, 201.3, 458.9, 10.53, 160.1, and the control IC50 ketoconazole was found to be less than 3.125 μM). These initial findings suggest that the above shortlisted steroidal samples may have in vivo activities as antioxidants, antifungal agents and as agents of drug-drug interactions via CYP3A4. The next step would be to obtain further analogues of the shortlist to see if potency can be increased and ideally full structural-activity relationships can be obtained to improve in silico testing. It is worthy to note that many of these compounds have limited solubility in the mostly aqueous assay conditions further conjugation with salts may also improve potencies.

Keywords: Steroids; Antioxidants; ABTS and DPPH Assays; CYP3A4; Drug-drug Interactions

×

References

  1. Mottram D and George A. “Anabolic steroids”. Best Practice and Research Clinical Endocrinology and Metabolism 1 (2000): 55-69.
  2. Rasheed A and Qasim M. “A review of natural steroids and their applications”. International Journal of Pharmaceutical Sciences and Research (2013).
  3. Oettel M. “The endocrine pharmacology of testosterone therapy in men”. Naturwissenschaften 2 (2004): 66-76.
  4. Lee K., et al. “Molecular mechanisms involved in progesterone receptor regulation of uterine function”. The Journal of Steroid Biochemistry and Molecular Biology1-5 (2006): 41-50
  5. Dai Y and Liu C. “Detection of 17 β-Estradiol in Environmental Samples and for Health Care Using a Single-Use, Cost-Effective Biosensor Based on Differential Pulse Voltammetry (DPV)”. Biosensors 4 (2017): 15.
  6. Li R., et al. “Defining ROS in Biology and Medicine” (2016).
  7. Kong Q., et al. “A threshold concept for cancer therapy”. Medical Hypotheses1 (2000): 29-35.
  8. Parsons B. “Antioxidants in Food: The Significance of Characterisation, Identification, Chemical and Biological Assays in Determining the Role of Antioxidants in Food”. Foods8 (2017): 68.
  9. Trolox, a Standard Antioxidant used for Antioxidant Capacity Assays | TCI Deutschland GmbH. Tcichemicals.com (2020).
  10. Carr A and Frei B. “Does vitamin C act as a pro‐oxidant under physiological conditions?”. The FASEB Journal9 (1999): 1007-1024.
  11. Ascorbic acid (2020).
  12. Su L., et al. “Total phenolic contents, chelating capacities, and radical-scavenging properties of black peppercorn, nutmeg, rosehip, cinnamon and oregano leaf”. Food Chemistry3 (2007): 990-997.
  13. Castillo J., et al. “Antioxidant and Radioprotective Effects of Olive Leaf Extract”. Olives and Olive Oil in Health and Disease Prevention (2010): 951-958.
  14. Kedare S and Singh R. “Genesis and development of DPPH method of antioxidant assay”. Journal of Food Science and Technology4 (2011): 412-422.
  15. Sara J., et al. “Evaluation of antioxidant activity of Malus domestica fruit extract from Kashan area”. African Journal of Agricultural Research20 (2015): 2136-2140.
  16. Yu L. “Free Radical Scavenging Properties of Conjugated Linoleic Acids”. Journal of Agricultural and Food Chemistry 7 (2001): 3452-3456.
  17. Hall P. “Role of Cytochromes P-450 in the Biosynthesis of Steroid Hormones”. Vitamins and Hormones (1985): 315-368.
  18. Preissner S., et al. “SuperCYP: a comprehensive database on Cytochrome P450 enzymes including a tool for analysis of CYP-drug interactions”. Nucleic Acids Research 38 (2009): D237-D243.
  19. Ince I., et al. “Developmental Changes in the Expression and Function of Cytochrome P450 3A Isoforms: Evidence from In Vitro and In Vivo Investigations”. Clinical Pharmacokinetics5 (2013): 333-345.
  20. CYP3A4 Enzyme | Everything You Need to Know About CYP3A4!. PharmaFactz (2018).
  21. Baldwin W and LeBlanc G. “In vivo biotransformation of testosterone by phase I and II detoxication enzymes and their modulation by 20-hydroxyecdysone in Daphnia magna”. Aquatic Toxicology 29 (1994): 103-117.
  22. Abel S., et al. “Cortisol metabolism in vitro—II. Species difference”. The Journal of Steroid Biochemistry and Molecular Biology5 (1993): 445-453.
  23. Kerlan V., et al. “Nature of cytochromes P450 involved in the 2-/4-hydroxylations of estradiol in human liver microsomes”. Biochemical Pharmacology9 (1992): 1745-1756.
  24. Alsanosi S., et al. “Pharmacokinetic Pharmacogenomics”. Handbook of Pharmacogenomics and Stratified Medicine (2014): 341-364.
  25. Novel Drug Approvals for 2018. U.S. Food and Drug Administration (2020).
  26. Bibi Z. “Role of cytochrome P450 in drug interactions”. Nutrition and Metabolism1 (2008): 27.
  27. What are some common medications classified as weak, moderate and strong inhibitors of CYP3A4? . Ebmconsult.com (2020).
  28. Ioannides C. Cytochrome P450. Cambridge: Royal Society of Chemistry (2008).
  29. Duina A., et al. “Budding Yeast for Budding Geneticists: A Primer on the Saccharomyces cerevisiae Model System”. Genetics1 (2014): 33-48.
  30. Lenasi H., et al. “Membrane-bound progesterone receptors coupled to G proteins in the fungus Rhizopus nigricans”. FEMS Microbiology Letters1 (2002): 97-101.
  31. Alam M., et al. “Review on in vivo and in vitro methods evaluation of antioxidant activity”. Saudi Pharmaceutical Journal 2 (2013): 143-152.
  32. Borrás C., et al. “Direct antioxidant and protective effect of estradiol on isolated mitochondria”. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease1 (2010): 205-211.
  33. ABUIN E., et al. “Uric acid reaction with dpph radicals at the micellar interface” (2020).
  34. Pisoschi A., et al. “Ascorbic Acid Determination in Commercial Fruit Juice Samples by Cyclic Voltammetry”. Journal of Automated Methods and Management in Chemistry (2008): 1-8.
  35. Fowler S and Zhang H. “In Vitro Evaluation of Reversible and Irreversible Cytochrome P450 Inhibition: Current Status on Methodologies and their Utility for Predicting Drug–Drug Interactions”. The AAPS Journal 2 (2008): 410-424.
  36. Krauser J., et al. “Cytochrome P450 3A4-catalyzed Testosterone 6β-Hydroxylation Stereochemistry, Kinetic Deuterium Isotope Effects, and Rate-limiting Steps”. Journal of Biological Chemistry20 (2005): 19496-19506.
  37. Harlow G and Halpert J. “Analysis of human cytochrome P450 3A4 cooperativity: Construction and characterization of a site-directed mutant that displays hyperbolic steroid hydroxylation kinetics”. Proceedings of the National Academy of Sciences12 (1998): 6636-6641.
  38. Stresser D., et al. “Highly selective inhibition of human cyp3a in vitro by azamulin and evidence that inhibition is irreversible”. Drug Metabolism and Disposition1 (2004): 105-112.
  39. Boetsch C., et al. “Effects of Cytochrome P450 3A4 Inhibitors—Ketoconazole and Erythromycin—on Bitopertin Pharmacokinetics and Comparison with Physiologically Based Modelling Predictions”. Clinical Pharmacokinetics2 (2015): 237-247.
  40. Dolan K., et al. “Antifungal Activity of Tamoxifen: In Vitro and In Vivo Activities and Mechanistic Characterization”. Antimicrobial Agents and Chemotherapy8 (2009): 3337-3346.
  41. Chen Y., et al. “A Mechanism for Tamoxifen-mediated Inhibition of Acidification”. Journal of Biological Chemistry 26 (1999): 18364-18373.
  42. Pubchem.ncbi.nlm.nih.gov (2020).
×

Citation

Citation: Dhvani H Kuntawala., et al. “Finding the Biological Properties from a Library of Steroids". Acta Scientific Biotechnology 1.11 (2020): 04-21.




Metrics

Acceptance rate36%
Acceptance to publication20-30 days

Indexed In


News and Events


  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for Upcoming Issue
    The last date for submission of articles for regular Issues is November 25, 2024.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of "Best Article of the Issue"
  • Welcoming Article Submission
    Acta Scientific delightfully welcomes active researchers for submission of articles towards the upcoming issue of respective journals.

Contact US