Structure of Nitisinone
CAS No.: 104206-65-7
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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
Nitisinone is an inhibitor of the enzyme 4-hydroxyphenylpyruvate dioxygenase.
Synonyms: NTBC; Nitisone; SC0735
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Clark, Callie ; Pokhrel, Menam ; Arthur, Benjamin ; Suresh, Pramita ; Ates, Ilayda ; Gibson, Justin , et al.
Abstract: We previously demonstrated lipid nanoparticle-mediated CRISPR-Cas9 gene editing to disrupt the gene encoding cytochrome P450 oxidoreductase (Cypor), combined with transient administration of acetaminophen (APAP), to repopulate the liver with healthy hepatocytes and rescue a phenylketonuria mouse model. This study aimed to investigate electroporation-mediated delivery of Cypor-targeting CRISPR-Cas9 ribonucleoproteins into wild-type hepatocytes, combined with liver engraftment under APAP treatment, as an in vivo selection approach in a mouse model of homozygous familial hypercholesterolemia (Ldlr−/−). Electroporation provides higher delivery efficiency compared to lipid nanoparticles. We observed engraftment levels up to 13% engraftment of electroporated Cypor-deficient hepatocytes with indels in the liver of Ldlr−/− mice after transient APAP administration, while negligible engraftment was observed in no-APAP controls (mean 9% and 2%, respectively, p = 0.0121). The engraftment of Cypor-deficient Ldlr+/+ hepatocytes was associated with reductions in LDL-cholesterol (18%) and triglycerides (52%) compared to the untransplanted control Ldlr−/− mice fed a Western diet for 5 weeks, but offered no protection from the development of diet-induced aortic root atherosclerosis or liver steatosis. While biochemical markers for liver damage normalized after discontinuation of APAP, we observed persistent lipid accumulation in the liver of Ldlr−/− mice grafted with Cypor-deficient Ldlr+/+ hepatocytes, likely stemming from the impact of Cypor deficiency on cholesterol clearance. Therefore, the combination of CRISPR-Cas9-mediated Cypor knockdown to induce clonal expansion of gene-edited hepatocytes using transient APAP administration is not a viable therapeutic strategy for familial hypercholesterolemia due to the essential role of Cypor in cholesterol metabolism. Unlike findings from phenylketonuria mouse model studies, the loss of Cypor function could not be compensated by unedited native hepatocytes in Ldlr−/− mice. Collectively, our results demonstrate that electroporation is a viable and informative approach for evaluating gene editing strategies for the treatment of inherited metabolic diseases that affect the liver. Our electroporation procedure revealed that a one-size-fits-all gene editing strategy may not be universally applicable for treating inherited metabolic liver disorders. Tailored gene editing and selection strategies may be needed for different liver disorders.
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Keywords: CRISPR-Cas9 ; gene editing ; hepatocyte transplantation ; electroporation ; liver research ; familial hypercholesterolemia ; atherosclerosis
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Polyploidy mitigates the impact of DNA damage while simultaneously bearing its burden
Hayashi, Kazuki ; Horisaka, Kisara ; Harada, Yoshiyuki ; Ogawa, Yuta ; Yamashita, Takako ; Kitano, Taku , et al.
Abstract: Polyploidy is frequently enhanced under pathological conditions, such as tissue injury and cancer in humans. Polyploidization is critically involved in cancer evolution, including cancer initiation and the acquisition of drug resistance. However, the effect of polyploidy on cell fate remains unclear. In this study, we explored the effects of polyploidization on cellular responses to DNA damage and cell cycle progression. Through various comparisons based on ploidy stratifications of cultured cells, we found that polyploidization and the accumulation of genomic DNA damage mutually induce each other, resulting in polyploid cells consistently containing more genomic DNA damage than diploid cells under both physiological and stress conditions. Notably, despite substantial DNA damage, polyploid cells demonstrated a higher tolerance to its impact, exhibiting delayed cell cycle arrest and reduced secretion of inflammatory cytokines associated with DNA damage-induced senescence. Consistently, in mice with ploidy tracing, hepatocytes with high ploidy appeared to potentially persist in the damaged liver, while being susceptible to DNA damage. Polyploidy acts as a reservoir of genomic damage by mitigating the impact of DNA damage, while simultaneously enhancing its accumulation.
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A novel enzymatic assay for the identification of 4-hydroxyphenylpyruvate dioxygenase modulators
Parkins, Andrew ; Pantouris, Georgios ;
Abstract: 4-hydroxyphenylpyruvate dioxygenase (HPPD) is a key enzyme involved in the pathogenesis of tyrosinemia III and cancer. Herein, we describe a spectroscopy-based assay to detect HPPD dioxygenase activity in the presence or absence of small-mol. modulators. We describe steps for transformation, expression, and purification of HPPD and preparation of the assay plate. We detail initiation and completion of the enzymic reaction followed by detection of remaining substrate in the form of enol-HPP/borate complex. This assay is applicable for high-throughput screening. For complete details on the use and execution of this protocol, please refer to Parkins et al.1
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Self-cleaving guide RNAs enable pharmacological selection of precise gene editing events in vivo
Tiyaboonchai, Amita ; Vonada, Anne ; Posey, Jeffrey ; Pelz, Carl ; Wakefield, Leslie ; Grompe, Markus
Abstract: Expression of guide RNAs in the CRISPR/Cas9 system typically requires the use of RNA polymerase III promoters, which are not cell-type specific. Flanking the gRNA with self-cleaving ribozyme motifs to create a self-cleaving gRNA overcomes this limitation. Here, we use self-cleaving gRNAs to create drug-selectable gene editing events in specific hepatocyte loci. A recombinant Adeno Associated Virus vector targeting the Albumin locus with a promoterless self-cleaving gRNA to create drug resistance is linked in cis with the therapeutic transgene. Gene expression of both are dependent on homologous recombination into the target locus. In vivo drug selection for the precisely edited hepatocytes allows >30-fold expansion of gene-edited cells and results in therapeutic levels of a human Factor 9 transgene. Importantly, self-cleaving gRNA expression is also achieved after targeting weak hepatocyte genes. We conclude that self-cleaving gRNAs are a powerful system to enable cell-type specific in vivo drug resistance for therapeutic gene editing applications.
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| CAS No. : | 104206-65-7 |
| Formula : | C14H10F3NO5 |
| M.W : | 329.23 |
| SMILES Code : | O=C1C(C(C2=CC=C(C(F)(F)F)C=C2[N+]([O-])=O)=O)C(CCC1)=O |
| Synonyms : |
NTBC; Nitisone; SC0735
|
| MDL No. : | MFCD01752192 |
| InChI Key : | OUBCNLGXQFSTLU-UHFFFAOYSA-N |
| Pubchem ID : | 115355 |
| GHS Pictogram: |
|
| Signal Word: | Warning |
| Hazard Statements: | H302+H312+H332-H315-H319-H335 |
| Precautionary Statements: | P261-P280-P305+P351+P338 |
| Description |
Nitisinone is an orally active, competitive, and reversible inhibitor of 4-hydroxyphenylpyruvate dioxygenase (4-HPPD) with an IC50 of 173 nM. It increases tyrosine levels in a dose-dependent manner and is utilized in research related to hereditary tyrosinemia type 1 (HT-1), a rare genetic disorder, and albinism[1].[2].[3].[4].
|
In Vitro:
| Concentration | Treated Time | Description | References |
| Saccharomyces cerevisiae | 4 µg per gram of yeast | 5.5 hours | Investigate the influence of nitisinone and its metabolites on yeast metabolism, finding that CHD caused a decrease in l-tyrosine, l-tryptophan, and l-phenylalanine concentrations by about 34%, 59% and 51%, respectively | Sci Rep. 2023 Jan 26;13(1):1473 |
In Vivo:
| Administration | Dosage | Frequency | Description | References |
| Zebrafish | Slc24a5 knockout zebrafish model | Added to E3 medium | 1.0, 2.5, 5, 10, 20, 50, 100, 150, and 200 μM | Drug water changed every 24 hours for 7 days | To evaluate the efficacy of nitisinone in improving melanin synthesis and pigmentation in the slc24a5 knockout zebrafish model, results showed no significant improvement in melanin content and pigmentation | Pigment Cell Melanoma Res. 2020 Jul;33(4):556-565 |
| Mice | Tyrc-h/c-h mice (OCA-1B model) and Tyrc-2J/c-2J mice (OCA-1A model) | Oral gavage | 4 mg/kg | Every other day for 1 month | To evaluate the effect of nitisinone on pigmentation in OCA-1B model mice. Results showed that nitisinone significantly increased fur and iris pigmentation in Tyrc-h/c-h mice but had no significant effect on Tyrc-2J/c-2J mice. | J Clin Invest. 2011 Oct;121(10):3914-23 |
| BALB/c mice | HGD−/− mice (AKU model) | Drinking water | 4 mg/L | Continuous for one week | To assess the effect of nitisinone-induced hypertyrosinaemia on monoamine neurotransmitters in a murine model of AKU. Results showed that nitisinone treatment significantly increased plasma tyrosine (9-fold) and decreased plasma homogentisic acid (9-fold), but no significant changes were observed in the distribution and intensity of dopamine, noradrenaline, and serotonin in brain tissue. | Metabolomics. 2019 Apr 29;15(5):68 |
| BALB/c Hgd−/− mice | Murine Model of Alkaptonuria (AKU) | Drinking water | 4 mg/L | Once daily for 6 days | To evaluate the impact of nitisinone on the CSF metabolome in a murine model of AKU, results showed significant increases in L-tyrosine and related metabolites post-treatment. | Metabolites. 2022 May 25;12(6):477 |
| Mice | Alkaptonuria (AKU) mouse model | Oral administration via drinking water | 4 mg/L (0.8 mg/kg) | Once daily for 7 days | To evaluate the effect of nitisinone on tyrosine metabolism pathway, results showed HGA was significantly decreased 8-fold and tyrosine increased 11-fold | J Inherit Metab Dis. 2020 Mar;43(2):259-268 |
| Mice | OCA3 mouse model | Oral gavage | 8 mg/kg | Every other day for 1 month | To evaluate the effect of Nitisinone on coat and ocular pigmentation in a mouse model of OCA3. Results showed that Nitisinone treatment significantly increased plasma tyrosine levels but did not improve coat or ocular pigmentation, with only a minimal increase in the number of pigmented melanosomes in the iris. | Invest Ophthalmol Vis Sci. 2018 Oct 1;59(12):4945-4952 |
Clinical Trial:
| NCT Number | Conditions | Phases | Recruitment | Completion Date | Locations |
| NCT01916382 | Alkaptonuria | PHASE3 | UNKNOWN | 2020-02-02 | Royal Liverpool Hospital, Live... More >>rpool, Merseyside, L7 8XP, United Kingdom Less << |
| NCT01828463 | Alkaptonuria | PHASE2 | COMPLETED | 2025-12-13 | Royal Liverpool Hospital, Live... More >>rpool, Merseyside, L7 8XP, United Kingdom Less << |
| NCT01838655 | Albinism|Vision Loss | PHASE1|PHASE2 | COMPLETED | 2017-02-07 | National Institutes of Health ... More >>Clinical Center, 9000 Rockville Pike, Bethesda, Maryland, 20892, United States Less << |
| NCT06227429 | Hereditary Tyrosinemia, Type I | RECRUITING | 2027-06-30 | Swedish Orphan Biovitrum Resea... More >>rch Site, Beijing, China|Swedish Orphan Biovitrum Research Site, Chongqing, China|Swedish Orphan Biovitrum Research Site, Hefei, China|Swedish Orphan Biovitrum Research Site, Wuhan, China Less << | |
| NCT01734889 | Hereditary Tyrosinemia, Type I | PHASE1 | COMPLETED | 2025-03-13 | Hopital Necker, Paris, France|... More >>Universit?tsklinikum Düsseldorf, Düsseldorf, Germany|Universit?tsklinikum Heidelberg, Heidelberg, Germany|Klinikum der Universit?t München, München, Germany|Birmingham Children's Hospital, Birmingham, United Kingdom|Evelina Children's Hospital, St Thomas' Hospital, London, United Kingdom|St Mary's Hospital, Manchester, United Kingdom Less << |
| NCT02750332 | Hereditary Tyrosinemia, Type I | PHASE1 | COMPLETED | 2025-01-16 | Bloemfontein Early Phase Clini... More >>cal Unit, PAREXEL International (South Africa), Bloemfontein, Free State, 9301, South Africa Less << |
| NCT02320084 | Hereditary Tyrosinemia, Type I | COMPLETED | 2019-09-30 | Swedish Orphan Biovitrum Inves... More >>tigational Site, Innsbruck, Austria|Swedish Orphan Biovitrum Investigational Site, Wien, Austria|Swedish Orphan Biovitrum Investigational Site, Antwerpen, Belgium|Swedish Orphan Biovitrum Investigational Site, Brussels, Belgium|Swedish Orphan Biovitrum Investigational Site, Leuven, Belgium|Swedish Orphan Biovitrum Investigational Site, Montegnée, Belgium|Swedish Orphan Biovitrum Investigational Site, Woluwe-Saint-Lambert, Belgium|Swedish Orphan Biovitrum Investigational Site, Prague, Czechia|Swedish Orphan Biovitrum Investigational Site, Copenhagen, Denmark|Swedish Orphan Biovitrum Investigational Site, Tampere, Finland|Swedish Orphan Biovitrum Investigational Site, Turku, Finland|Swedish Orphan Biovitrum Investigational Site, Angers, France|Swedish Oprhan Biovitrum Investigational Site, Lille, France|Swedish Orphan Biovitrum Investigational Site, Lyon, France|Swedish Orphan Biovitrum Investigational Site, Nantes, France|Swedish Oprhan Biovitrum Investigational Site, Paris, France|Swedish Orphan Biovitrum Investigational Site, Paris, France|Swedish Orphan Biovitrum Investigational Site, Talence, France|Swedish Orphan Biovitrum Investigational Site, Düsseldorf, Germany|Swedish Orphan Biovitrum Investigational Site, Erlangen, Germany|Swedish Orphan Biovitrum Investigational Site, Freiburg, Germany|Swedish Orphan Biovitrum Investigational Site, Hannover, Germany|Swedish Orphan Biovitrum Investigational Site, Heidelberg, Germany|Swedish Orphan Biovitrum Investigational Site, Leipzig, Germany|Swedish Orphan Biovitrum Investigational Site, Magdeburg, Germany|Swedish Orphan Biovitrum Investigational Site, München, Germany|Swedish Orphan Biovitrum Investigational Site, Reutlingen, Germany|Swedish Orphan Biovitrum Investigational Site, Budapest, Hungary|Swedish Oprhan Biovitrum Investigational Site, Szeged, Hungary|Swedish Orphan Biovitrum Investigational Site, Dublin, Ireland|Swedish Orphan Biovitrum Investigational Site, Bari, Italy|Swedish Orphan Biovitrum Investigational Site, Catania, Italy|Swedish Orphan Biovitrum Investigational Site, Firenze, Italy|Swedish Orphan Biovitrum Investigational Site, Milano, Italy|Swedish Orphan Biovitrum Investigational Site, Monza, Italy|Swedish Orphan Biovitrum Investigational Site, Napoli, Italy|Swedish Orphan Biovitrum Investigational Site, Padova, Italy|Swedish Orphan Biovitrum Investigational Site, Roma, Italy|Swedish Orphan Biovitrum Investigational Site, Saliceto, Italy|Swedish Orphan Biovitrum Investigational Site, Torino, Italy|Swedish Oprhan Biovitrum Investigational Site, Amsterdam, Netherlands|Swedish Orphan Biovitrum Investigational Site, Utrecht, Netherlands|Swedish Orphan Biovitrum Investigational Site, Fredrikstad, Norway|Swedish Orphan Biovitrum Investigational Site, Levanger, Norway|Swedish Oprhan Biovitrum Investigational Site, Oslo, Norway|Swedish Orphan Biovitrum Investigational Site, Oslo, Norway|Swedish Orphan Biovitrum Investigational Site, Troms?, Norway|Swedish Oprhan Biovitrum Investigational Site, Warsaw, Poland|Swedish Orphan Biovitrum Investigational Site, Warszawa, Poland|Swedish Orphan Biovitrum Investigational Site, Coimbra, Portugal|Swedish Orphan Biovitrum Investigational Site, Albacete, Spain|Swedish Orphan Biovitrum Investigational Site, Almeria, Spain|Swedish Orphan Biovitrum Investigational Site, Badalona, Spain|Swedish Orphan Biovitrum Investigational Site, Barakaldo, Spain|Swedish Orphan Biovitrum Investigational Site, Barcelona, Spain|Swedish Orphan Biovitrum Investigational Site, Las Palmas de Gran Canaria, Spain|Swedish Oprhan Biovitrum Investigational Site, Madrid, Spain|Swedish Orphan Biovitrum Investigational Site, Madrid, Spain|Swedish Orphan Biovitrum Investigational Site, Malaga, Spain|Swedish Orphan Biovitrum Investigational Site, Murcia, Spain|Swedish Orphan Biovitrum Investigational Site, Mérida, Spain|Swedish Orphan Biovitrum Investigational Site, Pamplona, Spain|Swedish Orphan Biovitrum Investigational Site, Santiago, Spain|Swedish Orphan Biovitrum Investigational Site, Sevilla, Spain|Swedish Orphan Biovitrum Investigational Site, Valencia, Spain|Swedish Orphan Biovitrum Investigational Site, G?teborg, Sweden|Swedish Orphan Biovitrum Investigational Site, Stockholm, 14186, Sweden|Swedish Orphan Biovitrum Investigational Site, Stockholm, Sweden|Swedish Orphan Biovitrum Investigational Site, Ume?, Sweden|Swedish Orphan Biovitrum Investigational Site, Uppsala, Sweden|Swedish Oprhan Biovitrum Investigational Site, Birmingham, United Kingdom|Swedish Orphan Biovitrum Investigational Site, Bradford, United Kingdom|Swedish Oprhan Biovitrum Investigational Site, London, United Kingdom|Swedish Orphan Biovitrum Investigational Site, London, United Kingdom|Swedish Oprhan Biovitrum Investigational Site, Manchester, United Kingdom|Swedish Orphan Biovitrum Investigational Site, Manchester, United Kingdom|Swedish Orphan Biovitrum Investigational Site, Sheffield, United Kingdom Less << | |
| NCT01390077 | Alkaptonuria | PHASE2|PHASE3 | COMPLETED | 2025-07-16 | University of California San D... More >>iego, La Jolla, California, 92093, United States Less << |
| NCT02323529 | Hereditary Tyrosinemia, Type I | PHASE3 | COMPLETED | 2025-09-15 | Swedish Orphan Biovitrum Inves... More >>tigational Site, Brussels, Belgium|Swedish Orphan Biovitrum Investigational Site, Copenhagen, Denmark|Swedish Orphan Biovitrum Investigational Site, Lyon, France|Swedish Orphan Biovitrum Investigational Site, Giessen, Germany|Swedish Orphan Biovitrum Investigational Site, Reutlingen, Germany|Swedish Orphan Biovitrum Investigational site, Gothenburg, Sweden Less << |
| NCT00004333 | Tyrosinemia, Type I | PHASE2 | COMPLETED | - | - |
| NCT00004443 | Tyrosinemia I | COMPLETED | 2025-09-06 | Oregon Health Sciences Univers... More >>ity, Portland, Oregon, 97201-3098, United States|University of Washington School of Medicine, Seattle, Washington, 98195, United States|Hopital Sainte Justine, Montreal, Quebec, H3T 1C5, Canada Less << |
| Bio Calculators | ||||
| Preparing Stock Solutions | ![]() |
1mg | 5mg | 10mg |
|
1 mM 5 mM 10 mM |
3.04mL 0.61mL 0.30mL |
15.19mL 3.04mL 1.52mL |
30.37mL 6.07mL 3.04mL |
|
| Dissolving Methods |
Please choose the appropriate dissolution scheme according to your animal administration guide.For the following dissolution schemes, clear stock solution should be prepared according to in vitro experiments, and then cosolvent should be added in turn:
in order to ensure the reliability of the experimental results, the clarified stock solution can be properly preserved according to the storage conditions; The working fluid for in vivo experiment is recommended to be prepared now and used on the same day; The percentage shown in front of the following solvent refers to the volume ratio of the solvent in the final solution; If precipitation or precipitation occurs in the preparation process, it can be assisted by heating and/or ultrasound.
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