Bcr-Abl

Specifically designed to interact with the ATP-binding site of BCR-ABL with a higher affinity than imatinib, nilotinib (AMN107), a selective Abl inhibitor, is also significantly more potent than imatinib (IC50<30 nM) and maintains activity against the majority of BCR-ABL point mutants that confer resistance to imatinib[1]. Nilotinib exhibits noteworthy antitumor efficacy against GIST xenograft lines and GIST cell lines resistant to imatinib; parent cell lines GK1C and GK3C display imatinib sensitivity with IC50 values of 4.59±0.97 µM and 11.15± 1.48 µM, respectively; imatinib-resistant cell lines GK1C-IR and GK3C-IR demonstrate Imatinib resistance with IC50 values of 11.74±0.17 µM (P<0.001) and 41.37±1.07 µM (P<0.001), respectively[2].

When given orally to BALB/cSLc-nu/nu mice with a GIST xenograft, nilotinib (oral gavage, 40 mg/kg, daily, 4 weeks) has anticancer effects that are same or greater[2]. Nilotinib reduces PDGFR α and β levels as well as apoptotic scores in the colon, while also having a strong healing effect on the macroscopic and microscopic pathologic scores and ensuring significant mucosal healing in the indomethacin-induced enterocolitis rat model[3].

Cell viability, cell cycle, and apoptosis analysis[1]
The trypan blue exclusion assay has been previously described, and was used to determine proliferation of cells cultured in the presence and absence of nilotinib, imatinib, or a combination of the 2 agents. Cell viability is reported as percentage of control (untreated) cells. Apoptosis of drug-treated cells was measured using the Annexin-V-Fluos Staining Kit, as previously described.

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Synergy studies[1]
For synergy studies, imatinib and nilotinib were added simultaneously at fixed ratios to imatinib-sensitive and imatinib-resistant BCR-ABL–expressing cells according to the method of Chou and Talalay. Cell viability was determined using the trypan blue exclusion assay. ED50 values were determined from the dose-response curves using graphic extrapolation. Specifically, (Y2 − Y1)/(X2 − X1) = (50 − Y1)(X50 − X1), where X50 = X1+ [(50 − Y1) * (X2 − X1)/(Y2 − Y1)] for linear x-axes and X50 = 10 (LOG10(C1)+(X − E1) * (LOG10(C2) − LOG10(C1)/(E2 − E1) for logarithmic x-axes. For calculation of the combination index, the following formula was used: (ICXa in mix/ICXa alone) + (ICXb in mix/ICXb alone). For the ICX value (nM), X is set to 25, 50, 75, or 90.

Animal/Disease Models: BALB/cSLc-nu/nu (nude) mice with GIST xenograft (GK1X, GK2X and GK3X)[2]
Doses: 40 mg/kg
Route of Administration: po (oral gavage); daily; 4 weeks
Experimental Results: Inhibited tumor growth by 69.6% in GK1X, 85.3% in GK2X and 47.5% in GK3X xenograft line.

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Male NCR-nude mice (5-6 weeks of age) were sublethally irradiated with a single fraction of 3 Gy, and approximately 3 hours later, a total of 800 000 cells was administered by tail-vein injection. Anesthetized mice were imaged and total body luminescence was measured as previously described. Baseline imaging 2 days after tumor cell inoculation was used to establish treatment cohorts with matched tumor burden. Cohorts of mice were treated with oral administration of vehicle (10% NMP, 90% PEG300), osmotic pump administration of 75 mg/kg imatinib, oral administration of 20 mg/kg per day nilotinib (diluted in 10% NMP, 90% PEG 300), or a combination of imatinib (75 mg/kg; osmotic pump) and nilotinib (20 mg/kg; oral gavage). Due to the significantly shorter half-life of imatinib in mice compared with humans, an alternative to continuous drug administration via the osmotic pump would entail twice daily intraperitoneal administration of imatinib, which has proved in our hands to be inefficient in terms of achieving maximum efficacy in mice. Treatment with vehicle and nilotinib was carried out for a total of 8 days; osmotic pumps were loaded with enough imatinib to allow up to 8 full days of treatment. Images were taken on days 2, 4, 5, and 7 after intravenous injection of 32D.p210-luc+ cells. On day 7 after intravenous injection, mice had received a total of 5 days of treatment with vehicle, nilotinib alone, imatinib alone, or the combination of nilotinib and imatinib. At the planned end of this study (9 days following the final imaging day), any remaining mice were killed, body and spleen weights were recorded, and tissues were preserved in 10% formalin for histopathologic analysis.[1]
Additional in vivo imaging studies were performed that included a variety of combinations of doses of nilotinib and imatinib, each administered alone and in combination to male NCR-nude mice (5-6 weeks of age). Drug formulations, treatments, and imaging were carried out as described above with some variations in experimental design (described in figure legends for Figures 6–7). Mice were administered the doses of nilotinib and imatinib, alone or in combination, at 20 mg/kg ± 50 mg/kg, 15 mg/kg ± 50 mg/kg, and then 15 mg/kg ± 75 mg/kg. Histopathologic analysis was then carried out.

[1]. Weisberg E, et al. Beneficial effects of combining nilotinib and imatinib in preclinical models of BCR-ABL+ leukemias. Blood. 2007 Mar 1;109(5):2112-20.
[2]. Sako H, et al. Antitumor effect of the tyrosine kinase inhibitor Nilotinib on gastrointestinal stromal tumor (GIST) and Imatinib-resistant GIST cells. PLoS One. 2014 Sep 15;9(9):e107613.
[3]. Dervis Hakim G, et al. Mucosal healing effect of nilotinib in indomethacin-induced enterocolitis: A rat model. World J Gastroenterol. 2015 Nov 28;21(44):12576-85.
[4]. Fujita KI, et al. Involvement of the Transporters P-Glycoprotein and Breast Cancer Resistance Protein in Dermal Distribution of the Multikinase Inhibitor Regorafenib and Its Active Metabolites. J Pharm Sci. 2017 Sep;106(9):2632-2641.
[5]. Meirson T, et al. Targeting invadopodia-mediated breast cancer metastasis by using ABL kinase inhibitors. Oncotarget. 2018 Apr 24;9(31):22158-22183.

Nilotinib is a member of (trifluoromethyl)benzenes, a member of pyrimidines, a member of pyridines, a member of imidazoles, a secondary amino compound and a secondary carboxamide. It has a role as an antineoplastic agent, a tyrosine kinase inhibitor and an anticoronaviral agent.
Nilotinib, also known as AMN107, is a tyrosine kinase inhibitor under investigation as a possible treatment for chronic myelogenous leukemia (CML). A Phase I clinical trial in 2006 showed that this drug was relatively safe and offered significant therapeutic benefits in cases of CML which were found to be resistant to treatment with imatinib (Gleevec), another tyrosine kinase inhibitor used as a first-line treatment for CML.
Nilotinib is a Kinase Inhibitor. The mechanism of action of nilotinib is as a Bcr-Abl Tyrosine Kinase Inhibitor, and Cytochrome P450 2C8 Inhibitor, and Cytochrome P450 2D6 Inhibitor, and Cytochrome P450 2B6 Inducer, and Cytochrome P450 2C8 Inducer, and UGT1A1 Inhibitor, and P-Glycoprotein Inhibitor.

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Nilotinib is a selective tyrosine kinase receptor inhibitor used in the therapy of chronic myelogenous leukemia. Nilotinib therapy is associated with transient elevations in serum aminotransferase levels and rare instances of clinically apparent acute liver injury.
Nilotinib is an orally bioavailable aminopyrimidine-derivative Bcr-Abl tyrosine kinase inhibitor with antineoplastic activity. Designed to overcome imatinib resistance, nilotinib binds to and stabilizes the inactive conformation of the kinase domain of the Abl protein of the Bcr-Abl fusion protein, resulting in the inhibition of the Bcr-Abl-mediated proliferation of Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML) cells. This agent also inhibits the receptor tyrosine kinases platelet-derived growth factor receptor (PDGF-R) and c-kit, a receptor tyrosine kinase mutated and constitutively activated in most gastrointestinal stromal tumors (GISTs). With a binding mode that is energetically more favorable than that of imatinib, nilotinib has been shown to have an approximately 20-fold increased potency in kinase and proliferation assays compared to imatinib.

C28H22F3N7O

529.52

529.18377

C, 63.51; H, 4.19; F, 10.76; N, 18.52; O, 3.02

641571-10-0

Nilotinib monohydrochloride monohydrate;923288-90-8;Nilotinib-d6;1268356-17-7;Nilotinib-d3;1215678-43-5;Nilotinib hydrochloride;923288-95-3; 641571-10-0; 923289-71-8 (hydrochloride dihydrate); 1277165-20-4 (dihydrochloride dihydrate)

644241

Typically exists as white to light yellow solids at room temperature

1.4±0.1 g/cm3

1.650

5.15

97.62

O=C(NC1=CC(C(F)(F)F)=CC(N2C=C(C)N=C2)=C1)C3=CC=C(C)C(NC4=NC=CC(C5=CC=CN=C5)=N4)=C3

HHZIURLSWUIHRB-UHFFFAOYSA-N

InChI=1S/C28H22F3N7O/c1-17-5-6-19(10-25(17)37-27-33-9-7-24(36-27)20-4-3-8-32-14-20)26(39)35-22-11-21(28(29,30)31)12-23(13-22)38-15-18(2)34-16-38/h3-16H,1-2H3,(H,35,39)(H,33,36,37)

4-methyl-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-{(4-(pyridin-3-yl)pyrimidin-2-yl)amino}benzamide

Nilotinib free base; AMN 107; AMN107; AMN-107; Tasigna; AMN107; AMN 107; AMN-107; nilotinibum; Nilotinib free base; Nilotinib; US brand name: Tasigna.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 0.5 mg/mL (0.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 5.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 2: 4% DMSO+30% PEG 300+5% Tween 80+ddH2O:3 mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)