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Systemic efficacy on Cryptosporidium parvum infection of aminoxanide (RM-5061), a new amino-acid ester thiazolide prodrug of tizoxanide

Published online by Cambridge University Press:  29 March 2021

El Hadji Diawara ,
Arnaud François ,
Andrew V. Stachulski ,
Romy Razakandrainibe ,
Damien Costa ,
Loïc Favennec ,
Jean-François Rossignol  and
Gilles Gargala Open the ORCID record for Gilles Gargala [Opens in a new window]
El Hadji Diawara
Affiliation:
Laboratoire de Parasitologie-Mycologie, Rouen University Hospital and EA7510, University of Rouen, Rouen, France
Arnaud François
Affiliation:
Laboratoire d'Anatomo-Pathologie, Rouen University Hospital, Rouen, France
Andrew V. Stachulski
Affiliation:
Robert Robinson Laboratories, Department of Chemistry, University of Liverpool, LiverpoolL69 7ZD, UK
Romy Razakandrainibe
Affiliation:
Laboratoire de Parasitologie-Mycologie, Rouen University Hospital and EA7510, University of Rouen, Rouen, France
Damien Costa
Affiliation:
Laboratoire de Parasitologie-Mycologie, Rouen University Hospital and EA7510, University of Rouen, Rouen, France
Loïc Favennec
Affiliation:
Laboratoire de Parasitologie-Mycologie, Rouen University Hospital and EA7510, University of Rouen, Rouen, France
Jean-François Rossignol
Affiliation:
Romark Laboratories, L.C., Tampa, FL33609, USA
Gilles Gargala*
Affiliation:
Laboratoire de Parasitologie-Mycologie, Rouen University Hospital and EA7510, University of Rouen, Rouen, France
*
Author for correspondence: Gilles Gargala, E-mail: [email protected]
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Abstract

Cryptosporidiosis is a gastrointestinal illness with profuse diarrhoea. Although there are no other Food and Drug Administration (FDA)-approved alternatives for the treatment of cryptosporidiosis, nitazoxanide (NTZ) can be qualified as partially effective. In immunosuppressed conditions, severe and/or disseminated cryptosporidiosis may occur and patients should be treated parenterally. To achieve the goal of developing parenteral treatment for cryptosporidiosis, the current study was undertaken to investigate the in vitro and in vivo anticryptosporidial activity of aminoxanide. This new l-tert-leucyl thiazolide is a soluble prodrug of tizoxanide (TIZ), the main metabolite of NTZ. Confirming the good efficacy of aminoxanide in Cryptosporidium parvum-infected HCT-8 cells with a 50% inhibitory concentration of 1.55 μm (±0.21), in immunosuppressed C. parvum-infected Mongolian gerbils (Meriones unguiculatus), a 5-day treatment with a daily intramuscular dose of 100 mg kg−1 aminoxanide resulted in a 72.5% oocyst excretion inhibition, statistically equivalent to 75.5% in gerbils treated with a 4-fold lower oral dose of NTZ. Cryptosporidium parvum-induced intestinal pathology and inflammation were improved. Aminoxanide provides an injectable form of TIZ that NTZ was unable to do and is a promising drug for which optimization of the formulation should be further explored. These results represent a first promising step towards the goal of developing a parenteral treatment for cryptosporidiosis.

Keywords

AminoxanidecryptosporidiosisCryptosporidium parvumdrug developmentnitazoxanide
Type
Research Article
Information
Parasitology , Volume 148 , Issue 8 , July 2021 , pp. 975 - 984
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Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press

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References

Adagu, IS, Nolder, D, Warhurst, DC and Rossignol, JF (2002) In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. Journal of Antimicrobial Chemotherapy 49, 103–111.CrossRefGoogle ScholarPubMed
Arnold, SLM, Choi, R, Hulverson, MA, Schaefer, DA, Vinayak, S, Vidadala, RSR, McCloskey, MC, Whitman, GR, Huang, W, Barrett, LK, Ojo, KK, Fan, E, Maly, DJ, Riggs, MW, Striepen, B and Van Voorhis, WC (2017) Necessity of bumped kinase inhibitor gastrointestinal exposure in treating Cryptosporidium infection. Journal of Infectious Diseases 216, 55–63.CrossRefGoogle ScholarPubMed
Barakat, FM, McDonald, V, Foster, GR, Tovey, MG and Korbel, DS (2009) Cryptosporidium parvum infection rapidly induces a protective innate immune response involving type I interferon. Journal of Infectious Diseases 200, 1548–1555.CrossRefGoogle ScholarPubMed
Barbot, L, Windsor, E, Rome, S, Tricottet, V, Reynes, M, Topouchian, A, Huneau, JF, Gobert, JG, Tome, D and Kapel, N (2003) Intestinal peptide transporter PepT1 is over-expressed during acute cryptosporidiosis in suckling rats as a result of both malnutrition and experimental parasite infection. Parasitology Research 89, 364–370.CrossRefGoogle ScholarPubMed
Bartelt, LA, Bolick, DT, Kolling, GL, Stebbins, E, Huston, CD, Guerrant, RL and Hoffman, PS (2018) Amixicile reduces severity of cryptosporidiosis but does not have in vitro activity against Cryptosporidium. Antimicrobial Agents and Chemotherapy 62, e00718–18.CrossRefGoogle Scholar
Broekhuysen, J, Stockis, A, Lins, RL, De Graeve, J and Rossignol, JF (2000) Nitazoxanide: pharmacokinetics and metabolism in man. International Journal of Clinical Pharmacology and Therapeutics 38, 387–394.CrossRefGoogle ScholarPubMed
Castellanos-Gonzalez, A, White, AC Jr, Ojo, KK, Vidadala, RS, Zhang, Z, Reid, MC, Fox, AM, Keyloun, KR, Rivas, K, Irani, A, Dann, SM, Fan, E, Dustin, J and Van Voorhis, WC (2013) A novel calcium-dependent protein kinase inhibitor as a lead compound for treating cryptosporidiosis. Journal of Infectious Diseases 208, 1342–1348.CrossRefGoogle ScholarPubMed
Chavez, MA and White, AC Jr (2018) Novel treatment strategies and drugs in development for cryptosporidiosis. Expert Review of Anti-Infective Therapy 16, 655–661.CrossRefGoogle ScholarPubMed
Checkley, W, White, AC Jr, Jaganath, D, Arrowood, MJ, Chalmers, RM, Chen, XM, Fayer, R, Griffiths, JK, Guerrant, RL, Hedstrom, L, Huston, CD, Kotloff, KL, Kang, G, Mead, JR, Miller, M, Petri, WA Jr, Priest, JW, Roos, DS, Striepen, B, Thompson, RC, Ward, HD, Van Voorhis, WA, Xiao, L, Zhu, G and Houpt, ER (2015) A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for Cryptosporidium. Lancet Infectious Diseases 15, 85–94.CrossRefGoogle ScholarPubMed
Chen, XM, Keithly, JS, Paya, CV and LaRusso, NF (2002) Cryptosporodiosis. New England Journal of Medicine 346, 1723–1731.CrossRefGoogle Scholar
Clifford, CP, Crook, DW, Conlon, CP, Fraise, AP, Day, DG and Peto, TE (1990) Impact of waterborne outbreak of cryptosporidiosis on AIDS and renal transplant patients. Lancet (London, England) 335, 1455–1456.CrossRefGoogle ScholarPubMed
Elazar, M, Liu, M, McKenna, SA, Liu, P, Gehrig, EA, Puglisi, JD, Rossignol, JF and Glenn, JS (2009) The anti-hepatitis C agent nitazoxanide induces phosphorylation of eukaryotic initiation factor 2alpha via protein kinase activated by double-stranded RNA activation. Gastroenterology 137, 1827–1835.CrossRefGoogle Scholar
Esposito, M, Stettler, R, Moores, SL, Pidathala, C, Müller, N, Stachulski, A, Berry, NG, Rossignol, JF and Hemphill, A (2005) In vitro efficacies of nitazoxanide and other thiazolides against Neospora caninum tachyzoites reveal antiparasitic activity independent of the nitro group. Antimicrobial Agents and Chemotherapy 49, 3715–3723.CrossRefGoogle ScholarPubMed
Funkhouser-Jones, LJ, Ravindran, S and Sibley, LD (2020) Defining stage-specific activity of potent new inhibitors of Cryptosporidium parvum growth in vitro. mBio 11, e00052–20.CrossRefGoogle ScholarPubMed
Gargala, G, Delaunay, A, Li, XD, Brasseur, P, Favennec, L and Ballet, JJ (2000) Efficacy of nitazoxanide, tizoxanide and tizoxanide-glucuronide against Cryptosporidium parvum development in sporozoite infected HCT-8 enterocytic cells. Journal of Antimicrobial Chemotherapy 46, 57–60.CrossRefGoogle ScholarPubMed
Gargala, G, Baishanbo, A, Favennec, L, François, A, Ballet, JJ and Rossignol, JF (2005) Inhibitory activities of epidermal growth factor receptor tyrosine kinase-targeted dihydroxyisoflavone and trihydroxydeoxybenzoin derivatives on Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum development. Antimicrobial Agents and Chemotherapy 49, 4628–4634.CrossRefGoogle ScholarPubMed
Gargala, G, Le Goff, L, Ballet, JJ, Favennec, L, Stachulski, AV and Rossignol, JF (2009) In vitro efficacy of nitro- and halogeno-thiazolide/thiadiazolide derivatives against Sarcocystis neurona. Veterinary Parasitology 162, 230–235.CrossRefGoogle ScholarPubMed
Gargala, G, François, A, Favennec, L and Rossignol, JF (2013) Activity of halogeno-thiazolides against Cryptosporidium parvum in experimentally infected immunosuppressed gerbils (Meriones unguiculatus). Antimicrobial Agents and Chemotherapy 57, 2821–2823.CrossRefGoogle Scholar
Gorla, SK, McNair, NN, Yang, G, Gao, S, Hu, M, Jala, VR, Haribabu, B, Striepen, B, Cuny, GD, Mead, JR and Hedstrom, L (2014) Validation of IMP dehydrogenase inhibitors in a mouse model of cryptosporidiosis. Antimicrobial Agents and Chemotherapy 58, 1603–1614.CrossRefGoogle Scholar
Griffiths, JK, Balakrishnan, R, Widmer, G and Tzipori, S (1998) Paromomycin and geneticin inhibit intracellular Cryptosporidium parvum without trafficking through the host cell cytoplasm: implications for drug delivery. Infection and Immunity 66, 3874–3883.CrossRefGoogle ScholarPubMed
Guo, A, Hu, P, Balimane, PV, Leibach, FH and Sinko, PJ (1999) Interactions of a nonpeptidic drug, valacyclovir, with the human intestinal peptide transporter (hPEPT1) expressed in a mammalian cell line. Journal of Pharmacological Experimental Therapeutics 289, 448–454.Google Scholar
Heine, J (1982) Eine einfache nachweismethod fur kryptosporidien in kot. Zentralblatt für Veterinärmedizin Reihe B 29, 324–327.CrossRefGoogle Scholar
Hemphill, A, Mueller, J and Esposito, M (2006) Nitazoxanide, a broad-spectrum thiazolide anti-infective agent for the treatment of gastrointestinal infections. Expert Opinion on Pharmacotherapy 7, 953–964.CrossRefGoogle ScholarPubMed
Hunter, PR and Nichols, G (2002) Epidemiology and clinical features of Cryptosporidium infection in immunocompromised patients. Clinical Microbiology Reviews 15, 145–154.CrossRefGoogle ScholarPubMed
Huston, CD, Spangenberg, T, Burrows, J, Willis, P, Wells, TN and Van Voorhis, W (2015) A proposed target product profile and developmental cascade for new cryptosporidiosis treatments. PLoS Neglected Tropical Diseases 9, e0003987.CrossRefGoogle ScholarPubMed
Jumani, RS, Bessoff, K, Love, MS, Miller, P, Stebbins, EE, Teixeira, JE, Campbell, MA, Meyers, MJ, Zambriski, JA, Nunez, V, Woods, AK, McNamara, CW and Huston, CD (2018) A novel piperazine-based drug lead for cryptosporidiosis from the medicines for malaria venture open-access malaria box. Antimicrobial Agents and Chemotherapy 62, e01505–e01517.CrossRefGoogle ScholarPubMed
Kutukculer, N, Moratto, D, Aydinok, Y, Lougaris, V, Aksoylar, S, Plebani, A, Genel, F and Notarangelo, LD (2003) Disseminated Cryptosporidium infection in an infant with hyper-IgM syndrome caused by CD40 deficiency. Journal of Pediatrics 142, 194–196.CrossRefGoogle Scholar
Lee, S, Harwood, M, Girouard, D, Meyers, MJ, Campbell, MA, Beamer, G and Tzipori, S (2017) The therapeutic efficacy of azithromycin and nitazoxanide in the acute pig model of Cryptosporidium hominis. PLoS One 12, e0185906.CrossRefGoogle ScholarPubMed
Li, XD, Brasseur, P, Agnamey, P, Leméteil, D, Favennec, L, Ballet, JJ and Rossignol, JF (2003) Long-lasting anticryptosporidial activity of nitazoxanide in an immunosuppressed rat model. Folia Parasitologica (Praha) 50, 19–22.CrossRefGoogle Scholar
McClure, S and Palma, K (1999) Treatment of equine protozoal myeloencephalitis with nitazoxanide. Journal of Equine Veterinary Science 19, 639–641.CrossRefGoogle Scholar
Mor, SM, Tumwine, JK, Ndeezi, G, Srinivasan, MG, Kaddu-Mulindwa, DH, Tzipori, S and Griffiths, JK (2010) Respiratory cryptosporidiosis in HIV-seronegative children in Uganda: potential for respiratory transmission. Clinical Infectious Diseases 50, 1366–1372.CrossRefGoogle ScholarPubMed
Müller, J, Sidler, D, Nachbur, U, Wastling, J, Brunner, T and Hemphill, A (2008) Thiazolides inhibit growth and induce glutathione-S-transferase Pi (GSTP1)-dependent cell death in human colon cancer cells. International Journal of Cancer 123, 1797–1806.CrossRefGoogle ScholarPubMed
Reagan-Shaw, S, Nihal, M and Ahmad, N (2007) Dose translation from animal to human studies revisited. FASEB Journal 22, 659–661.CrossRefGoogle ScholarPubMed
Rossignol, JF (2014) Nitazoxanide: a first-in-class broad-spectrum antiviral agent. Antiviral Research 110, 94–103.CrossRefGoogle ScholarPubMed
Rossignol, JF and Stachulski, AV (1999) Syntheses and antibacterial activities of tizoxanide, an N-(nitrothiazolyl)salicylamide, and its O-aryl glucuronide. Journal of Chemical Research (Synopses) 23, 44–45. doi: 10.1177/174751989902300128CrossRefGoogle Scholar
Rossignol, JF, Ayoub, A and Ayers, MS (2001) Treatment of diarrhea caused by Cryptosporidium parvum: a prospective randomized, double blind, placebo-controlled study of nitazoxanide. Journal of Infectious Diseases 184, 103–106.CrossRefGoogle ScholarPubMed
Sparks, H, Nair, G, Castellanos-Gonzalez, A and White, AC Jr (2015) Treatment of Cryptosporidium: what we know, gaps, and the way forward. Current Tropical Medicine Reports 2, 181–187.CrossRefGoogle ScholarPubMed
Sponseller, JK, Griffiths, JK and Tzipori, S (2014) The evolution of respiratory cryptosporidiosis: evidence for transmission by inhalation. Clinical Microbiology Reviews 27, 575–586.CrossRefGoogle ScholarPubMed
Stachulski, AV, Swift, K, Cooper, M, Reynolds, S, Norton, D, Slonecker, SD and Rossignol, JF (2017) Synthesis and pre-clinical studies of new amino-acid ester thiazolide prodrugs. European Journal of Medicinal Chemistry 126, 154–159.CrossRefGoogle ScholarPubMed
The Medical Letter (2013) Drugs for parasitic infections. Treatment Guidelines from the Medical Letter 11(Suppl), e2–31.Google Scholar
Tilmanis, D, van Baalen, C, Oh, DY, Rossignol, JF and Hurt, AC (2017) The susceptibility of circulating human influenza viruses to tizoxanide, the active metabolite of nitazoxanide. Antiviral Research 147, 142–148.CrossRefGoogle ScholarPubMed
Topouchian, A, Huneau, JF, Barbot, L, Rome, S, Gobert, JG, Tomé, D and Kapel, N (2003) Evidence for the absence of an intestinal adaptive mechanism to compensate for C. parvum-induced amino acid malabsorption in suckling rats. Parasitology Research 91, 197–203.CrossRefGoogle Scholar
Trabattoni, D, Gnudi, F, Ibba, SV, Saulle, I, Agostini, S, Masetti, M, Biasin, M, Rossignol, JF and Clerici, M (2016) Thiazolides elicit anti-viral innate immunity and reduce HIV. Scientific Reports 6, 27148.CrossRefGoogle ScholarPubMed
Tzipori, S and Griffiths, JK (1998) Natural history and biology of Cryptosporidium parvum. Advances in Parasitology 40, 5–36.CrossRefGoogle ScholarPubMed