Skip to main content

Advertisement

SpringerLink
Log in

Preclinical Evaluation of the Short-Term Toxicity of 4-(N)-Docosahexaenoyl 2´, 2´- Difluorodeoxycytidine (DHA-dFdC)

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose

This study was designed to test the short-term toxicity of DHA-dFdC in a mouse model and its efficacy in a mouse model of leukemia at or below its repeat-dose maximum tolerated dose (RD-MTD).

Method

A repeat-dose dose-ranging toxicity study was designed to determine the tolerability of DHA-dFdC when administered to DBA/2 mice by intravenous (i.v.) injection on a repeat-dose schedule (i.e. injections on days 0, 3, 7, 10, and 13). In order to determine the effect of a lethal dose of DHA-dFdC, mice were injected i.v. with three doses of DHA-dFdC at 100 mg/kg on days 0, 3, and 5 (i.e. a lethal-RD). The body weight of mice was recorded two or three times a week. At the end of the study, major organs (i.e. heart, liver, spleen, kidneys, lung, and pancreas) of mice that received the lethal-RD or RD-MTD were weighed, and blood samples were collected for analyses. Finally, DHA-dFdC was i.v. injected into DBA/2 mice with syngeneic L1210 mouse leukemia cells to evaluate its efficacy at or below RD-MTD.

Results

The RD-MTD of DHA-dFdC is 50 mg/kg. At 100 mg/kg, a lethal-RD, DHA-dFdC decreases the weights of mouse spleen and liver and significantly affected certain blood parameters (i.e. white blood cells, lymphocytes, eosinophils, and neutrophil segmented). At or below its RD-MTD, DHA-dFdC significantly prolonged the survival of L1210 leukemia-bearing mice.

Conclusion

DHA-dFdC has dose-dependent toxicity, affecting mainly spleen at a lethal-RD. At or below its RD-MTD, DHA-dFdC is effective against leukemia in a mouse model.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

ALP:

Alkaline phosphatase

ALT:

Alanine transaminase

AST:

Aspartate transaminase

BUN:

Blood urea nitrogen

CPK:

Creatine phosphokinase

dFdC:

2`, 2`-difluorodeoxycytidine HCl

DHA:

Docosahexaenoic acid

DHA-dFdC:

4-(N)-docosahexaenoyl 2´, 2´-difluorodeoxycytidine

HCT:

Hematocrit value

HGB:

Hemoglobin level

i.p.:

Intraperitoneally

i.v.:

Intravenous

ILS:

Increase in life span

Lethal-RD:

Lethal-repeat dose

PSCs:

Pancreatic stellate cells

PUFA:

Polyunsaturated fatty acid

RBC:

Red blood cell count

RD-MTD:

Repeat-dose maximum tolerated dose

T/U:

The ratio of the mean survival time of treated group to the median survival time of the untreated group

WBC:

White blood cells

References

  1. Carmichael J, Fink U, Russell RC, Spittle MF, Harris AL, Spiessi G, Blatter J. Phase II study of gemcitabine in patients with advanced pancreatic cancer. Br J Cancer. 1996;73(1):101–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Burris 3rd HA, Moore MJ, Andersen J, Green MR, Rothenberg ML, Modiano MR, Cripps MC, Portenoy RK, Storniolo AM, Tarassoff P, Nelson R, Dorr FA, Stephens CD, Von Hoff DD. Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: a randomized trial. J Clin Oncol. 1997;15(6):2403–13.

    Article  CAS  PubMed  Google Scholar 

  3. Hoang T, Kim K, Jaslowski A, Koch P, Beatty P, McGovern J, Quisumbing M, Shapiro G, Witte R, Schiller JH. Phase II study of second-line gemcitabine in sensitive or refractory small cell lung cancer. Lung Cancer. 2003;42(1):97–102.

    Article  PubMed  Google Scholar 

  4. Pfisterer J, Plante M, Vergote I, du Bois A, Hirte H, Lacave AJ, Wagner U, Stahle A, Stuart G, Kimmig R, Olbricht S, Le T, Emerich J, Kuhn W, Bentley J, Jackisch C, Luck HJ, Rochon J, Zimmermann AH, Eisenhauer E. Gemcitabine plus carboplatin compared with carboplatin in patients with platinum-sensitive recurrent ovarian cancer: an intergroup trial of the AGO-OVAR, the NCIC CTG, and the EORTC GCG. J Clin Oncol. 2006;24(29):4699–707.

    Article  CAS  PubMed  Google Scholar 

  5. Yardley DA. Gemcitabine plus paclitaxel in breast cancer. Semin Oncol. 2005;32(4 Suppl 6):S14–21.

    Article  CAS  PubMed  Google Scholar 

  6. Wong A, Soo RA, Yong WP, Innocenti F. Clinical pharmacology and pharmacogenetics of gemcitabine. Drug Metab Rev. 2009;41(2):77–88.

    Article  CAS  PubMed  Google Scholar 

  7. de Sousa CL, Monteiro G. Gemcitabine: metabolism and molecular mechanisms of action, sensitivity and chemoresistance in pancreatic cancer. Eur J Pharmacol. 2014;741:8–16.

    Article  Google Scholar 

  8. Casper ES, Green MR, Kelsen DP, Heelan RT, Brown TD, Flombaum CD, Trochanowski B, Tarassoff PG. Phase II trial of gemcitabine (2,2'-difluorodeoxycytidine) in patients with adenocarcinoma of the pancreas. Investig New Drugs. 1994;12(1):29–34.

    Article  CAS  Google Scholar 

  9. Storniolo AM, Allerheiligen SR, Pearce HL. Preclinical, pharmacologic, and phase I studies of gemcitabine. Semin Oncol. 1997;24(2 Suppl 7):S7-2–7.

    Google Scholar 

  10. Merendino N, Costantini L, Manzi L, Molinari R, D'Eliseo D, Velotti F. Dietary omega −3 polyunsaturated fatty acid DHA: a potential adjuvant in the treatment of cancer. Biomed Res Int. 2013;2013:310186.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Chamras H, Ardashian A, Heber D, Glaspy JA. Fatty acid modulation of MCF-7 human breast cancer cell proliferation, apoptosis and differentiation. J Nutr Biochem. 2002;13(12):711–6.

    Article  CAS  PubMed  Google Scholar 

  12. Serini S, Piccioni E, Merendino N, Calviello G. Dietary polyunsaturated fatty acids as inducers of apoptosis: implications for cancer. Apoptosis. 2009;14(2):135–52.

    Article  CAS  PubMed  Google Scholar 

  13. Spencer L, Mann C, Metcalfe M, Webb M, Pollard C, Spencer D, Berry D, Steward W, Dennison A. The effect of omega-3 FAs on tumour angiogenesis and their therapeutic potential. Eur J Cancer. 2009;45(12):2077–86.

    Article  CAS  PubMed  Google Scholar 

  14. Horia E, Watkins BA. Complementary actions of docosahexaenoic acid and genistein on COX-2, PGE2 and invasiveness in MDA-MB-231 breast cancer cells. Carcinogenesis. 2007;28(4):809–15.

    Article  CAS  PubMed  Google Scholar 

  15. D'Eliseo D, Manzi L, Merendino N, Velotti F. Docosahexaenoic acid inhibits invasion of human RT112 urinary bladder and PT45 pancreatic carcinoma cells via down-modulation of granzyme B expression. J Nutr Biochem. 2012;23(5):452–7.

    Article  PubMed  Google Scholar 

  16. Li S, Qin J, Tian C, Cao J, Fida G, Wang Z, Chen H, Qian Z, Chen WR, Gu Y. The targeting mechanism of DHA ligand and its conjugate with gemcitabine for the enhanced tumor therapy. Oncotarget. 2014;5(11):3622–35.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Arterburn LM, Boswell KD, Koskelo E, Kassner SL, Kelly C, Kyle DJ (2000) A combined subchronic (90-day) toxicity and neurotoxicity study of a single-cell source of docosahexaenoic acid triglyceride (DHASCO oil). Food Chem Toxicol. 38:35–49.

  18. Nelson GJ, Schmidt PC, Bartolini GL, Kelley DS, Kyle D. The effect of dietary docosahexaenoic acid on plasma lipoproteins and tissue fatty acid composition in humans. Lipids. 1997;32(11):1137–46.

    Article  CAS  PubMed  Google Scholar 

  19. Ryan AS, Keske MA, Hoffman JP, Nelson EB. Clinical overview of algal-docosahexaenoic acid: effects on triglyceride levels and other cardiovascular risk factors. Am J Ther. 2009;16(2):183–92.

    Article  PubMed  Google Scholar 

  20. Nelson GJ, Schmidt PS, Bartolini GL, Kelley DS, Kyle D. The effect of dietary docosahexaenoic acid on platelet function, platelet fatty acid composition, and blood coagulation in humans. Lipids. 1997;32(11):1129–36.

    Article  CAS  PubMed  Google Scholar 

  21. Park Y, Harris W. EPA, but not DHA, decreases mean platelet volume in normal subjects. Lipids. 2002;37(10):941–6.

    Article  CAS  PubMed  Google Scholar 

  22. Kelley DS, Taylor PC, Nelson GJ, Mackey BE. Dietary docosahexaenoic acid and immunocompetence in young healthy men. Lipids. 1998;33(6):559–66.

    Article  CAS  PubMed  Google Scholar 

  23. Thies F, Nebe-von-Caron G, Powell JR, Yaqoob P, Newsholme EA, Calder PC. Dietary supplementation with gamma-linolenic acid or fish oil decreases T lymphocyte proliferation in healthy older humans. J Nutr. 2001;131(7):1918–27.

    CAS  PubMed  Google Scholar 

  24. He K, Xun P, Brasky TM, Gammon MD, Stevens J, White E. Types of fish consumed and fish preparation methods in relation to pancreatic cancer incidence: the VITAL cohort study. Am J Epidemiol. 2013;177(2):152–60.

    Article  PubMed  Google Scholar 

  25. Zheng JS, Hu XJ, Zhao YM, Yang J, Li D. Intake of fish and marine n-3 polyunsaturated fatty acids and risk of breast cancer: meta-analysis of data from 21 independent prospective cohort studies. BMJ. 2013;346:f3706.

    Article  PubMed  Google Scholar 

  26. Arshad A, Chung WY, Steward W, Metcalfe MS, Dennison AR. Reduction in circulating pro-angiogenic and pro-inflammatory factors is related to improved outcomes in patients with advanced pancreatic cancer treated with gemcitabine and intravenous omega-3 fish oil. HPB: Off J Int Hepato Pancreato Biliary Assoc. 2013;15(6):428–32.

    Article  Google Scholar 

  27. Arshad A, Isherwood J, Mann C, Cooke J, Pollard C, Runau F, Morgan B, Steward W, Metcalfe M, Dennison A. Intravenous omega-3 fatty acids plus gemcitabine: potential to improve response and quality of life in advanced pancreatic cancer. JPEN J Parenteral Enter Nutr 2015.

  28. Camargo Cde Q, Mocellin MC, Pastore Silva Jde A, Fabre ME, Nunes EA. Trindade EB. Fish oil supplementation during chemotherapy increases posterior time to tumor progression in colorectal cancer. Nutr Cancer. 2016;68(1):70–6.

    Article  PubMed  Google Scholar 

  29. Murphy RA, Mourtzakis M, Chu QS, Baracos VE, Reiman T, Mazurak VC. Supplementation with fish oil increases first-line chemotherapy efficacy in patients with advanced nonsmall cell lung cancer. Cancer. 2011;117(16):3774–80.

    Article  CAS  PubMed  Google Scholar 

  30. Colomer R, Moreno-Nogueira JM, Garcia-Luna PP, Garcia-Peris P, Garcia-de-Lorenzo A, Zarazaga A, Quecedo L, del Llano J, Usan L, Casimiro C. N-3 fatty acids, cancer and cachexia: a systematic review of the literature. Br J Nutr. 2007;97(5):823–31.

    Article  CAS  PubMed  Google Scholar 

  31. Heller AR, Rossel T, Gottschlich B, Tiebel O, Menschikowski M, Litz RJ, Zimmermann T, Koch T. Omega-3 fatty acids improve liver and pancreas function in postoperative cancer patients. Int J Cancer. 2004;111(4):611–6.

    Article  CAS  PubMed  Google Scholar 

  32. Naguib YW, Lansakara PD, Lashinger LM, Rodriguez BL, Valdes S, Niu M, Aldayel AM, Peng L, Hursting SD, Cui Z. Synthesis, characterization, and in vitro and in vivo evaluations of 4-(N)-docosahexaenoyl 2', 2'-difluorodeoxycytidine with potent and broad-Spectrum antitumor activity. Neoplasia. 2016;18(1):33–48.

    Article  CAS  PubMed  Google Scholar 

  33. Montgomery CA. Oncological and toxicological research: alleviation and control of pain and distress in laboratory animals. Cancer Bull. 1990;42:230–7.

    Google Scholar 

  34. NRC. The Guide for the Care and Use of Laboratory Animals. Washington, D.C: National Academies Press; 2011.

  35. Gianasi E, Wasil M, Evagorou EG, Keddle A, Wilson G, Duncan R. HPMA copolymer platinates as novel antitumour agents: in vitro properties, pharmacokinetics and antitumour activity in vivo. Eur J Cancer. 1999;35(6):994–1002.

    Article  CAS  PubMed  Google Scholar 

  36. Andersson L, Davies J, Duncan R, Ferruti P, Ford J, Kneller S, Mendichi R, Pasut G, Schiavon O, Summerford C, Tirk A, Veronese FM, Vincenzi V, Wu G. Poly(ethylene glycol)-poly(ester-carbonate) block copolymers carrying PEG-peptidyl-doxorubicin pendant side chains: synthesis and evaluation as anticancer conjugates. Biomacromolecules. 2005;6(2):914–26.

    Article  CAS  PubMed  Google Scholar 

  37. Reddy LH, Marque PE, Dubernet C, Mouelhi SL, Desmaele D, Couvreur P. Preclinical toxicology (subacute and acute) and efficacy of a new squalenoyl gemcitabine anticancer nanomedicine. J Pharmacol Exp Ther. 2008;325(2):484–90.

    Article  CAS  PubMed  Google Scholar 

  38. Raabe BM, Artwohl JE, Purcell JE, Lovaglio J, Fortman JD. Effects of weekly blood collection in C57BL/6 mice. J Am Assoc Lab Anim Sci. 2011;50(5):680–5.

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Oliver SAG, Smith JE, Kaneko JJ. Erythrocyte Structure and Function. In: Douglas J, Weiss KJW, editor. Schalm's Veterinary Hematology. Ames: John Wiley & Sons; 2010. p. 123–130.

  40. Hall R, Everds N. Principles of Clinical Pathology for Toxicology Studies. In. Principles and Methods of Toxicology, Fifth Edition: CRC Press, Boca Raton; 2007. p. 1317–1358.

  41. MacNeill A. Clinical Biochemistry of Domestic Animals, 6th Edition by Editors: J. Jerry Kaneko, John W. Harvey, and Michael L. Bruss. Veterinary Clinical Pathology. 2009;38(4):545.

  42. Department of Health and Human Services UFaDA. Substances affirmed as generally recognized as safe: menhaden oil. In. Washington: Food and Drug Administration, HHS. 1997. p. 30751–30757.

  43. Klek S. Omega-3 Fatty Acids in Modern Parenteral Nutrition: A Review of the Current Evidence. J Clin Med. 2016;5(3).

  44. Haqq J, Howells LM, Garcea G, Dennison AR. Targeting pancreatic cancer using a combination of gemcitabine with the omega-3 polyunsaturated fatty acid emulsion, Lipidem. Mol Nutr Food Res. 2016;60(6):1437–47.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Pharmaceutics Division, College of Pharmacy, The University of Texas at Austin, Austin, Texas, 78712, USA

    Solange Valdes, Youssef W. Naguib & Zhengrong Cui

  2. Department of Veterinary Sciences, Michale E. Keeling Center for Comparative Medicine and Research, University of Texas M.D. Anderson Cancer Center, Bastrop, Texas, 78602, USA

    Rick A. Finch & Wallace B. Baze

  3. Department of Nutritional Sciences, College of Natural Sciences, The University of Texas at Austin, Austin, Texas, 78712, USA

    Christopher A. Jolly

Authors
  1. Solange Valdes
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Youssef W. Naguib
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rick A. Finch
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wallace B. Baze
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christopher A. Jolly
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhengrong Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhengrong Cui.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Valdes, S., Naguib, Y.W., Finch, R.A. et al. Preclinical Evaluation of the Short-Term Toxicity of 4-(N)-Docosahexaenoyl 2´, 2´- Difluorodeoxycytidine (DHA-dFdC). Pharm Res 34, 1224–1232 (2017). https://doi.org/10.1007/s11095-017-2139-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-017-2139-x

Key words

Search

Navigation