| Size | Price | Stock | Qty |
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| 10mg |
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| Targets |
IC50: Nampt[1]
OT-82 targets nicotinamide phosphoribosyltransferase (NAMPT), a key enzyme in the NAD+ biosynthesis pathway. NAMPT catalyzes the conversion of nicotinamide to nicotinamide mononucleotide (NMN), the rate-limiting step in the salvage pathway for NAD+ production. By inhibiting NAMPT, OT-82 disrupts NAD+ metabolism, which is vital for cellular energy regulation, DNA repair, and survival. Cancer cells, particularly those of hematopoietic origin, have a strong dependence on NAD+ biosynthesis, making them selectively vulnerable to NAMPT inhibition. |
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| ln Vitro |
For a duration of 72 hours, OT-82 (0.0001-10 μM) exhibits tissue-specific (HP versus non-HP) cytotoxicity. Its IC50 values are 2.11 nM, 2.70 nM, 1.05 nM, and 1.36 nM for HP cell lines MV4–11, U937, RS4;11, HEL92.1.7, and PER485 cell proliferation, respectively. With IC50 values of 37.92 nM, 29.52 nM, 15.67 nM, and 7.95 nM, respectively, it also inhibits the development of nonHP cell lines MCF-7, U87, HT29, and H1299[1]. OT-82 exhibits cytotoxicity that is cancer-selective (tumor vs. normal). The IC50 values for AML and ALL donors, respectively, are 31 nM and 7.10 nM, indicating that it is more sensitive to BMMNC from leukemia patients. In healthy donors, the IC50 value for BMMNC is 62.69 nM[1]. In MV4-11 cells, OT-82 (0.001-10 μM; 48 hours) generates dose-dependent decreases in cellular NAD and ATP concentrations as well as inhibits the activity of recombinant NAMPT[1]. (0.01-100 nM; 48 hours) causes the mitochondrial membrane of MV4–11 cells to depolarize, an increase in the percentage of cells with sub-G1 DNA content, and the activation of caspase-3[1].
OT-82 exhibits potent in vitro activity by inhibiting NAMPT and inducing cell death in hematologic cancer cell lines with low-nanomolar IC50 values. In MV4-11 cells, OT-82 (0.01-100 nM) causes depolarization of the mitochondrial membrane. The compound exerts its antileukemic action by depleting NAD+ and ATP, inhibiting the NAD+-requiring DNA damage repair enzyme PARP-1, increasing mitochondrial ROS levels, and inducing DNA damage, culminating in apoptosis induction. OT-82 is selectively toxic to hematopoietic cells and induces cell death in a NAD+-dependent manner. |
| ln Vivo |
In the SC xenograft model of Burkitt's lymphoma, OT-82 (oral gavage; 20 or 40 mg/kg; 3 weeks) treatment increases survival to 100% and 56% at 40 or 20 mg/kg, respectively, after treatment discontinuation[1].
In vivo, OT-82 has demonstrated anticancer activity in preclinical models. Upon oral administration, the NAMPT inhibitor OT-82 impairs tumor growth and extends survival in mice bearing EWS xenografts. Its selective toxicity towards hematopoietic cells is attributed to their dependence on NAD+ biosynthesis for rapid proliferation. OT-82 is a promising anticancer drug candidate that targets the strong dependence of hematological malignancies on NAD biosynthesis. |
| Enzyme Assay |
In vitro enzyme assays for OT-82 typically measure its inhibition of NAMPT enzymatic activity. These assays use recombinant NAMPT and a substrate such as nicotinamide, with the production of NMN measured via coupled enzymatic reactions or HPLC. The IC50 is determined from dose-response curves. Additionally, cellular NAD+ and ATP levels are quantified using standard biochemical kits to confirm target engagement and downstream metabolic effects. PARP-1 activity and ROS levels can also be measured to assess the compound's mechanism of action.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: HP cell lines (MV4–11, U937, RS4;11, HEL92.1.7, PER485) Non-HP cell lines (MCF-7, U87, HT29, H1299) Tested Concentrations: 0.0001 μM-10 μM Incubation Duration: 72 hrs (hours) Experimental Results: Was against human cell lines derived from hematological malignancies (HP) with IC50 values ranging from 1.10 nM to 5.86 nM, and was against non-HP cancers with IC50 ranging from 1.10 nM to 37.92 nM[1]. [1] Cell Types: MV4–11 cells Tested Concentrations: 0.01-100 nM Incubation Duration: 48 hrs (hours) Experimental Results: demonstrated hallmarks of apoptotic cell death In vitro cellular assays for OT-82 are conducted in hematologic cancer cell lines such as MV4-11. Cells are treated with varying concentrations of OT-82 (e.g., 0.01-100 nM), and cell viability is measured using MTT or CellTiter-Glo assays. Mitochondrial membrane potential is assessed using fluorescent dyes like JC-1. Apoptosis is evaluated by measuring caspase activation, PARP cleavage, or Annexin V staining. NAD+ and ATP levels are quantified, and DNA damage is assessed via γ-H2AX staining or comet assays. |
| Animal Protocol |
Animal/Disease Models: SC xenograft model of Burkitt's lymphoma in SCID (severe combined immunodeficient) mouse[1]
Doses: 20 or 40 mg/kg Route of Administration: po (oral gavage); 3 weeks Experimental Results: Potently inhibited tumor growth of multiple myeloma mouse model. In vivo animal experiments with OT-82 are conducted in mouse xenograft models, such as EWS (Ewing sarcoma) xenografts. Tumor-bearing mice are administered OT-82 orally at various doses. Tumor growth is monitored over time, and survival is recorded. Pharmacodynamic endpoints include measurement of tumor NAD+ and ATP levels, assessment of DNA damage (γ-H2AX), and analysis of apoptosis markers (caspase-3, PARP). Pharmacokinetic parameters such as plasma exposure and bioavailability are also assessed. |
| ADME/Pharmacokinetics |
OT-82 is an orally active compound with favorable pharmacokinetic properties. It has a molecular weight of 424.5 g/mol and a molecular formula of C26H21FN4O. The compound is typically stored as a powder at -20°C (3 years) or 4°C (2 years), and in solution at -80°C (6 months) or -20°C (1 month). Its half-life, bioavailability, and tissue distribution have been characterized in preclinical studies to support its efficacy in xenograft models.
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| Toxicity/Toxicokinetics |
Comprehensive toxicological data for OT-82 are primarily derived from preclinical studies. The compound is intended for research use only and is not approved for human therapeutic use. In animal models, OT-82 has been shown to be well-tolerated at efficacious doses, but full toxicological characterization is not extensively published. Standard safety assessments include cytotoxicity assays in relevant cell lines and monitoring of body weight, clinical signs, and organ histopathology in animal studies.
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| References | |
| Additional Infomation |
OT-82, a NAMPT inhibitor, is a small molecule inhibitor with high oral bioavailability that inhibits nicotinamide adenine dinucleotide (NAD) synthase—nicotinamide phosphoribosyltransferase (NAMPT; NAMPRTase)—and possesses potential antitumor activity. After oral administration, OT-82 binds to NAMPT and inhibits its activity. This depletes intracellular NAD and inhibits NAD-dependent enzymes, both essential for rapid cell proliferation; ultimately leading to the death of NAMPT-overexpressing cancer cells. NAMPT is an enzyme responsible for maintaining the intracellular NAD pool, playing a crucial role in cellular metabolic regulation and exhibiting cytokine-like activity. NAMPT is overexpressed in various cancers and metabolic disorders; tumor cells depend on NAMPT activity to acquire NAD.
OT-82 is a potent, selective, and orally active NAMPT inhibitor being developed for the treatment of hematological malignancies. Its mechanism involves NAD+ depletion, leading to energy crisis, PARP-1 inhibition, ROS increase, and DNA damage-induced apoptosis. It has demonstrated preclinical efficacy in leukemia models. The compound is available in high purity (≥98%) for research applications. Further clinical development would require additional toxicity and efficacy studies. |
| Molecular Formula |
C26H21FN4O
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|---|---|
| Molecular Weight |
424.469548940659
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| Exact Mass |
424.169
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| CAS # |
1800487-55-1
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| Related CAS # |
1800487-55-1;OT-82 HCl;
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| PubChem CID |
118203189
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| Appearance |
White to light yellow solid powder
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| LogP |
4.5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
32
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| Complexity |
659
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC1C=CC(=CC=1)C#CC1C=C(C=CC=1C1C=CN=CC=1)C(NCCCC1C=NNC=1)=O
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| InChi Key |
CEPAXRIKSUXHHB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H21FN4O/c27-24-8-4-19(5-9-24)3-6-22-16-23(7-10-25(22)21-11-14-28-15-12-21)26(32)29-13-1-2-20-17-30-31-18-20/h4-5,7-12,14-18H,1-2,13H2,(H,29,32)(H,30,31)
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| Chemical Name |
3-[2-(4-fluorophenyl)ethynyl]-N-[3-(1H-pyrazol-4-yl)propyl]-4-pyridin-4-ylbenzamide
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| Synonyms |
OT82 OT 82
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~235.59 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.90 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.90 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.90 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3559 mL | 11.7794 mL | 23.5588 mL | |
| 5 mM | 0.4712 mL | 2.3559 mL | 4.7118 mL | |
| 10 mM | 0.2356 mL | 1.1779 mL | 2.3559 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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