Small molecule metalloprotease inhibitor with in vitro, ex vivo and in vivo efficacy against botulinum neurotoxin serotype A

The botulinum neurotoxins (BoNTs) produced by Clostridium botulinum are the most poisonous protein substances known. The neutralizing antibodies against botulinum neurotoxin can effectively prevent and cure the toxicosis. Using purified Hc fragments of botulinum neurotoxin serotype A (BoNT/A-Hc) as antigen, 2 specific neutralizing antibodies mapping different epitopes were selected from a fully synthetic human antibody library. The 2 antibodies can effectively inhibit the binding between BoNT/A-Hc and differentiated PC-12 cells in vitro, and the neutralization was evaluated in vivo. Although no single mAb completely protected mice from toxin, they both could prolong time to death when challenged with 20 LD 50s (50% lethal doses) of BoNT/A. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, suggesting their high neutralizing potency in vivo . The results would lead to further production of neutralizing antibody drugs against BoNT/A. It also proved that it was a quick method to obtain human therapeutic antibodies by selecting from the fully synthetic human antibody phage display library. ( Journal of Biomolecular Screening 2009:991-998)

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Erratum to “A first-in-class CDK4 inhibitor demonstrates in vitro, ex-vivo and in vivo efficacy against ovarian cancer” [Gynecologic Oncology 159 (2020) 827–838]

Gynecologic Oncology ◽

10.1016/j.ygyno.2021.07.018 ◽

2021 ◽

Author(s):

Laychiluh Bantie ◽

Solomon Tadesse ◽

Jimma Likisa ◽

Mingfeng Yu ◽

Benjamin Noll ◽

...

Keyword(s):

Ovarian Cancer ◽

Ex Vivo ◽

Gynecologic Oncology ◽

In Vivo Efficacy ◽

Cdk4 Inhibitor

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Computer-Aided Lead Optimization: Improved Small-Molecule Inhibitor of the Zinc Endopeptidase of Botulinum Neurotoxin Serotype A

PLoS ONE ◽

10.1371/journal.pone.0000761 ◽

2007 ◽

Vol 2 (8) ◽

pp. e761 ◽

Cited By ~ 35

Author(s):

Jing Tang ◽

Jewn Giew Park ◽

Charles B. Millard ◽

James J. Schmidt ◽

Yuan-Ping Pang

Keyword(s):

Small Molecule ◽

Botulinum Neurotoxin ◽

Small Molecule Inhibitor ◽

Lead Optimization ◽

Serotype A ◽

Molecule Inhibitor ◽

Computer Aided ◽

Botulinum Neurotoxin Serotype A

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Novel 2,4-diaminopyrimidines bearing tetrahydronaphthalenyl moiety against anaplastic lymphoma kinase (ALK): Synthesis, in vitro, ex vivo, and in vivo efficacy studies

Bioorganic & Medicinal Chemistry Letters ◽

10.1016/j.bmcl.2016.02.052 ◽

2016 ◽

Vol 26 (7) ◽

pp. 1720-1725 ◽

Cited By ~ 2

Author(s):

Dawn Song ◽

Minji Lee ◽

Chi Hoon Park ◽

Sunjoo Ahn ◽

Chang Soo Yun ◽

...

Keyword(s):

Anaplastic Lymphoma Kinase ◽

Ex Vivo ◽

In Vivo Efficacy ◽

Anaplastic Lymphoma ◽

Efficacy Studies

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Potency comparison in in vitro, ex vivo, and in vivo assays of commercially available botulinum neurotoxin serotypes A1, B1, and F1

Toxicon ◽

10.1016/j.toxicon.2018.11.061 ◽

2018 ◽

Vol 156 ◽

pp. S25

Author(s):

Sarah Donald ◽

Mark Elliott ◽

Bryony Gray ◽

Fraser Hornby ◽

Agnieszka Lewandowska ◽

...

Keyword(s):

Botulinum Neurotoxin ◽

Ex Vivo ◽

In Vivo Assays

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Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models

Pharmaceutics ◽

10.3390/pharmaceutics12070611 ◽

2020 ◽

Vol 12 (7) ◽

pp. 611 ◽

Cited By ~ 1

Author(s):

Jaakko Itkonen ◽

Ada Annala ◽

Shirin Tavakoli ◽

Blanca Arango-Gonzalez ◽

Marius Ueffing ◽

...

Keyword(s):

Ex Vivo ◽

Disease Model ◽

Neural Retina ◽

Biologically Active ◽

Target Cells ◽

Therapeutic Effects ◽

Duration Of Action ◽

In Vivo Efficacy

Ciliary neurotrophic factor (CNTF) is one of the most studied neuroprotective agents with acknowledged potential in treating diseases of the posterior eye segment. Although its efficacy and mechanisms of action in the retina have been studied extensively, it is still not comprehensively understood which retinal cells mediate the therapeutic effects of CNTF. As with therapeutic proteins in general, it is poorly elucidated whether exogenous CNTF administered into the vitreous can enter and distribute into the retina and hence reach potentially responsive target cells. Here, we have characterized our purified recombinant human CNTF (rhCNTF), studied the protein’s in vitro bioactivity in a cell-based assay, and evaluated the thermodynamic and oligomeric status of the protein during storage. Biological activity of rhCNTF was further evaluated in vivo in an animal model of retinal degeneration. The retinal penetration and distribution of rhCNTF after 24 h was studied utilizing two ex vivo retina models. Based on our characterization findings, our rhCNTF is correctly folded and biologically active. Moreover, based on initial screening and subsequent follow-up, we identified two buffers in which rhCNTF retains its stability during storage. Whereas rhCNTF did not show photoreceptor preservative effect or improve the function of photoreceptors in vivo, this could possibly be due to the used disease model or the short duration of action with a single intravitreal injection of rhCNTF. On the other hand, the lack of in vivo efficacy was shown to not be due to distribution limitations; permeation into the retina was observed in both retinal explant models as in 24 h rhCNTF penetrated the inner limiting membrane, and being mostly observed in the ganglion cell layer, distributed to different layers of the neural retina. As rhCNTF can reach deeper retinal layers, in general, having direct effects on resident CNTF-responsive target cells is plausible.

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Pure Botulinum Neurotoxin Is Absorbed from the Stomach and Small Intestine and Produces Peripheral Neuromuscular Blockade

Infection and Immunity ◽

10.1128/iai.67.9.4708-4712.1999 ◽

1999 ◽

Vol 67 (9) ◽

pp. 4708-4712 ◽

Cited By ~ 84

Author(s):

Andrew B. Maksymowych ◽

Marco Reinhard ◽

Carl J. Malizio ◽

Michael C. Goodnough ◽

Eric A. Johnson ◽

...

Keyword(s):

Small Intestine ◽

Mouse Model ◽

Heavy Chain ◽

Clostridium Botulinum ◽

General Circulation ◽

Botulinum Neurotoxin ◽

Serotype A ◽

Relative Absorption ◽

Botulinum Neurotoxin Serotype A

ABSTRACT Clostridium botulinum serotype A produces a neurotoxin composed of a 100-kDa heavy chain and a 50-kDa light chain linked by a disulfide bond. This neurotoxin is part of a ca. 900-kDa complex, formed by noncovalent association with a single nontoxin, nonhemagglutinin subunit and a family of hemagglutinating proteins. Previous work has suggested, although never conclusively demonstrated, that neurotoxin alone cannot survive passage through the stomach and/or cannot be absorbed from the gut without the involvement of auxiliary proteins in the complex. Therefore, this study compared the relative absorption and toxicity of three preparations of neurotoxin in an in vivo mouse model. Equimolar amounts of serotype A complex with hemagglutinins, complex without hemagglutinins, and purified neurotoxin were surgically introduced into the stomach or into the small intestine. In some experiments, movement of neurotoxin from the site of administration was restricted by ligation of the pylorus. Comparison of relative toxicities demonstrated that at adequate doses, complex with hemagglutinins, complex without hemagglutinins, and pure neurotoxin can be absorbed from the stomach. The potency of neurotoxin in complex was greater than that of pure neurotoxin, but the magnitude of this difference diminished as the dosage of neurotoxin increased. Qualitatively similar results were obtained when complex with hemagglutinins, complex without hemagglutinins, and pure neurotoxin were placed directly into the intestine. This work establishes that pure botulinum neurotoxin serotype A is toxic when administered orally. This means that pure neurotoxin does not require hemagglutinins or other auxiliary proteins for absorption from the gastrointestinal system into the general circulation.

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