Therapeutic Efficacy of Bumped Kinase Inhibitor 1369 in a Pig Model of Acute Diarrhea Caused by Cryptosporidium hominis

Lee, Sangun; Ginese, Melanie; Beamer, Gillian; Danz, Hillary; Girouard, Donald; Chapman-Bonofiglio, Susan; Lee, Minhee; Hulverson, Matthew A.; Choi, Ryan; Whitman, Grant R.; Ojo, Kayode K.; Arnold, Samuel; Voorhis, Wesley C. Van; Tzipori, Saul

doi:10.1128/aac.00147-18

Cited by 38 publications

(47 citation statements)

References 27 publications

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“…BKI-1369 and BKI-1770 are lead compounds for the treatment of diarrhea caused by cryptosporidiosis in calves [15,17]. In addition, excellent BKI-1369 efficacy was demonstrated against Cystisospora suis infection in piglets [18], and in a pig model of acute diarrhea caused by Cryptosporidium hominis ([16].…”

Section: Discussionmentioning

confidence: 99%

Comparative assessment of the effects of bumped kinase inhibitors on early zebrafish embryo development and pregnancy in mice

Anghel

Winzer

Imhof

et al. 2020

Preprint

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Bumped kinase inhibitors (BKIs) are effective against a variety of apicomplexan parasites. Fifteen BKIs with promising in vitro efficacy against Neospora caninum tachyzoites, low cytotoxicity in mammalian cells, and no toxic effects in non-pregnant BALB/c mice, were assessed in pregnant mice. Drugs were emulsified in corn oil and applied by gavage for 5 days. Five BKIs did not affect pregnancy, 5 BKIs exhibited 15-35% of neonatal mortality, and 5 compounds caused strong effects (infertility, abortion, stillbirth and pup mortality). Additionally, the impact of these compounds on zebrafish (Danio rerio) embryo development was assessed by exposing freshly fertilized eggs to 0.2-50μM of BKIs and microscopical monitoring of embryo development in a blinded manner during 4 days. We propose an algorithm that includes quantification of malformations and embryo deaths, and established a scoring system that allows to calculate an impact score (Si) that indicates at which concentrations BKIs visibly affect zebrafish embryo development. Comparison of the two models showed that for 9 compounds no clear correlation between Si and pregnancy outcome was visible. However, those 3 BKIs affecting zebrafish embryos only at high concentrations (40μM or higher) did not impair mouse pregnancy at all, and those 3 compounds that inhibited zebrafish embryo development already at 0.2μM showed detrimental effects in the pregnancy model. Thus, the zebrafish embryo development test has a limited predictive value to foresee pregnancy outcome in BKI-treated mice. We conclude, that maternal health-related factors such as cardiovascular, pharmacokinetic and/or bioavailability properties also contribute to BKI-pregnancy effects.

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Section: Discussionmentioning

confidence: 99%

Comparative assessment of the effects of bumped kinase inhibitors on early zebrafish embryo development and pregnancy in mice

Anghel

Winzer

Imhof

et al. 2020

Preprint

Self Cite

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“…There has been considerable recent investment in Cryptosporidium drug discovery and there are now several classes of compounds of lead, late lead, and preclinical status. These include bumped kinase inhibitors that target Cryptosporidium calcium-dependant protein kinase 1 (CDPK1) and are effective in several in vivo models of cryptosporidiosis, including gnotobiotic piglets and calf models, which showed a reduction in oocyst shedding and clinical signs [53,54]. Compounds that target tRNA synthetases, enzymes that charge tRNAs with their cognate amino acid for use in protein synthesis, have also proven effective against Cryptosporidium [55].…”

Section: Advances In Drug Discoverymentioning

confidence: 99%

A One Health Approach to Tackle Cryptosporidiosis

Innes

Chalmers

Wells

et al. 2020

Trends in Parasitology

142

103

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“…While Giardia cysts made in mice excyst well, cysts made in vitro do not (57). Toxoplasma forms walled oocysts in cats, while Cryptosporidium forms oocysts in cows, mice, or gnotobiotic pigs (58-60). Oomycetes (water molds that cause potato blight) also have cellulose and chitin in their walls, but the amount of chitin is very small, and β- 1,3-glucan is also present (61).…”

Section: Discussionmentioning

confidence: 99%

“…MBP-fusion constructs were prepared by cloning the cDNA of a representative Luke(2) lectin (60 to 843-bp CDS of ACA1_377670) and a representative Jonah(1) lectin (70 to 1599-bp CDS of ACA1_164810) without their signal sequences into pMAL-p2x vector (New England Biolabs) for periplasmic expression in BL21-CodonPlus(DE3)-RIPL (Agilent Technologies, Lexington, MA) (34). For the MBP-fusion, the Leo CDS without the signal sequence (67 to 564-bp of ACA1_074730) was codon optimized (GenScript) and cloned into pMAL-p2x vector.…”

Section: Methodsmentioning

confidence: 99%

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

Samuelson

Magistrado-Coxen

Aqeel

et al. 2018

Preprint

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18Acanthamoeba castellanii, cause of keratitis and blindness, is an emerging pathogen because 19 of its association with contact lens use. The cyst wall contributes to pathogenesis as cysts are 20 resistant to sterilizing reagents in lens solutions and to antibiotics applied to the eye. We used 21 transmission electron microscopy, as well as structured illumination microscopy and probes for 22 cellulose and chitin, to show that purified cyst walls of A. castellanii retain an outer ectocyst 23 layer, an inner endocyst layer, and conical ostioles that connect the layers. Mass spectrometry 24 showed candidate cyst wall proteins are dominated by three families of lectins (named here 25 Luke, Leo, and Jonah), each of which binds to microcrystalline cellulose and to a lesser degree 26 chitin. A Jonah lectin, which has one choice-of-anchor A (CAA) domain, localizes to the 27 ectocyst layer of mature cyst walls. Luke lectins, which have two or three carbohydrate-binding 28 modules (CBM49), localize to the endocyst layer and ostioles. A Leo lectin, which has two 29 domains with eight Cys residues each (8-Cys), also localizes to the endocyst layer and 30 ostioles. In summary, the most abundant A. castellanii cyst wall proteins are three sets of 31 lectins, which have carbohydrate-binding modules that are conserved (CBM49s of Luke), 32 newly characterized (CAA of Jonah), or unique to Acanthamoebae (8-Cys of Leo). Despite 33 their lack of common ancestry, Luke and Leo lectins both localize to the endocyst layer and 34 ostioles, while the Jonah lectin localizes to the ectocyst layer. 35IMPORTANCE 36 Fifty years ago, investigators identified cellulose in the Acanthamoeba cyst wall, which has an 37 outer ectocyst layer, an inner endocyst layer, and conical ostioles that connect the layers. 38Here we show cyst walls also contain chitin and three large sets of cellulose-and chitin-binding 39 lectins, which have distinct localizations. The Acanthamoeba cyst wall, therefore, is more 40 complicated than cyst walls of Entamoeba and Giardia (causes of dysentery and diarrhea, 41 respectively), which have a single layer, a single glycopolymer, and small sets of one or two 42 lectins. In contrast, the Acanthamoeba cyst wall is far simpler than the walls of fungi and 43 plants, which have multiple layers, numerous glycopolymers, and hundreds of proteins. In 44 addition to providing a better understanding of the cell biology and biochemistry of the 45 Acanthamoeba cyst wall, these studies may lead to diagnostic antibodies that bind to cyst wall 46 proteins and/or therapeutics that target chitin. 47 100 file S1) (41). The GST-AcCBM49 expression construct was designed to replicate that used to 101 determine the carbohydrate binding properties of SlCBM49, which is a C-terminal 102 carbohydrate-binding module of the Solanum lycopersicum (tomato) cellulase SlGH9C (42). In 103 both intact cysts and purified cyst walls, GST-AcCBM49 predominantly labeled the ectocyst 104 layer, WGA highlighted the ostioles, and CFW stained the endocyst...

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Therapeutic Efficacy of Bumped Kinase Inhibitor 1369 in a Pig Model of Acute Diarrhea Caused by Cryptosporidium hominis

Cited by 38 publications

References 27 publications

Comparative assessment of the effects of bumped kinase inhibitors on early zebrafish embryo development and pregnancy in mice

Comparative assessment of the effects of bumped kinase inhibitors on early zebrafish embryo development and pregnancy in mice

A One Health Approach to Tackle Cryptosporidiosis

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

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