MEK1 (IC50 = 0.92 nM); MEK2 (IC50 = 1.8 nM)

GSK1120212 has an IC50 range of 0.92 nM to 3.4 nM and inhibits the phosphorylation of MBP regardless of the isotypes of Raf and MEK. c-Raf, B-Raf, ERK1 and ERK2 are not inhibited by GSK1120212's kinase activity. Furthermore, the other 98 kinases are not significantly inhibited by GSK1120212 in a significant way. The human colorectal cancer cell lines are effectively inhibited by GSK1120212. The cells with the highest sensitivity to GSK1120212 have IC50 values of 0.48 nM and 0.52 nM, respectively, and are known to have a constitutively active B-Raf mutant in HT-29 and COLO205. With an IC50 range of 2.2–174 nM, the cell lines with the K-Ras mutation exhibit a wide range of sensitivity to GSK1120212. The wild-type gene in both B-Raf and K-Ras is present in COLO320 DM cells, which are resistant to GSK1120212 even at 10 μM. All sensitive cell lines experience cell-cycle arrest at the G1 phase after a 24-hour treatment with GSK1120212. p15INK4b and/or p27KIP1 are consistently upregulated by GSK1120212 treatment in the majority of colorectal cancer cell lines. ERK phosphorylation by GSK1120212 is inhibited in all susceptible cell lines. Both HT-29 and COLO205 cells experience apoptosis induction from GSK1120212; however, COLO205 cells are more vulnerable to this induction than HT-29 cells are. [1] Peripheral blood mononuclear cells (PBMCs) cannot produce tumor necrosis factor or interleukin-6 because GSK1120212 inhibits this process. [2]

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LPS-induced ERK1/2 phosphorylation and proinflammatory cytokine production were inhibited by Trametinib/JTP-74057 [2]
JTP-74057 is a potent MEK1/2 inhibitor that specifically inhibits MEK1/2, with an IC50 value of about 2 nM. Because it is well known that LPS induces ERK1/2 phosphorylation in monocytes via the COT/Tpl2–MEK1/2 pathway, we examined inhibitory activity of JTP-74057 against the phosphorylation of ERK1/2 in LPS-stimulated human, mouse, or rat PBMCs. The ERK1/2 phosphorylation, which was rapidly phosphorylated upon LPS stimulation within 30 min, was completely inhibited with 10 nM of JTP-74057 in all species, demonstrating that there is no species difference in the inhibitory activity of this compound (Fig. 1). The same results were obtained using human PBMCs from different donors, and in the repetitive experiments for mouse and rat PBMCs (data not shown). Since activation of the MEK–ERK pathway has been reported in the synovial tissue of RA patients, and this activation leads to production of proinflammatory cytokines such as TNF-α and IL-6, we next examined the effect of the MEK1/2 inhibitor on cytokine production from LPS-stimulated hPBMCs. As shown in Fig. 2, consistent with inhibitory activity of MEK1/2, 10 nM of JTP-74057 inhibited production of TNF-α by about 10% of the control. IL-6 production was also inhibited; however, the maximum inhibition was about 50% of the control, even at 100 nM of the compound, implying that there might be pathway(s) to activate IL-6 production other than the MEK–ERK pathway.

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Differential effects of Trametinib/JTP-74057 and leflunomide on the production of anti-CII antibody and the reactivation of CII-reactive T cells [2]
To investigate whether MEK1/2 inhibition affects autoantibody production, anti-CII IgG in serum was measured by ELISA on day 35. Leflunomide inhibited the elevation of anti-CII IgG in a dose-dependent manner. On the other hand, JTP-74057 did not affect anti-CII IgG production, even at the highest dose (Fig. 6a), suggesting that MEK1/2 does not play a role in autoantibody production. Next, we examined the effect of the MEK1/2 inhibitor on reactivation of antigen-specific memory T cells in the CIA mouse. Lymph node cells were collected from non-drug-treated mice with CIA 5 days after the second CII immunization, and then re-stimulated with heat-degradated type II collagen in vitro in the presence or absence of test drugs. Two days later, proliferation of LN cells was assessed by [3H]thymidine incorporation. JTP-74057 inhibited the proliferation of LN cells upon CII stimulation (Fig. 6b), implying that the suppressive effect of the MEK1/2 inhibitor on CIA development was, at least in part, due to blocking the reactivation of antigen-specific memory T cells. A77 1726, the active metabolite of leflunomide, showed a minimal effect on proliferation of LN cells (Fig. 6b). These results clearly demonstrated that the MEK inhibitor has different disease-modifying activity from leflunomide.

GSK1120212 can effectively stop the growth of the HT-29 xenograft when given orally at doses of 0.3 mg/kg or 1 mg/kg once daily for 14 days. At doses of 1 mg/kg, the tumor growth is almost entirely stopped. A single oral dose of 1 mg/kg GSK1120212 completely inhibits the phosphorylation of ERK1/2 in the tissues of established tumors, and after 14 days of treatment, the levels of the proteins p15INK4b and p27KIP1 are both increased. Tumor regression is seen in the COLO205 xenograft model even at a dose of 0.3 mg/kg. Four out of six mice receiving a dose of 1 mg/kg experience a complete regression, in which the tumor has regressed to the point where its volume is no longer detectable. [1] Adjuvant-induced arthritis (AIA) and type II collagen-induced arthritis (CIA) in Lewis rats or DBA1/J mice, respectively, are almost completely suppressed after administration of GSK1120212 at 0.1 mg/kg. [2]

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The effect of JTP-74057/Trametinib on rat adjuvant-induced arthritis model [2]
To confirm the pharmacological effects of the MEK1/2 inhibitor on inflammatory arthritis development, we first employed the rat adjuvant-induced arthritis (AIA) model, which is widely used as a model for RA. Male Lewis rats were intradermally administered with M. tuberculosis-containing adjuvant at the tail base on day 0, and then the volume of hind paws was monitored. On day 21, joint destruction of hind paws was radiographically evaluated. JTP-74057 was orally administered once a day from day 0. Leflunomide was used as a reference drug. As shown in Fig. 3, Trametinib/JTP-74057 dramatically blocked hind paw swelling in a dose-dependent manner, and 0.1 mg/kg of JTP-74057 showed comparable efficacy to 10 mg/kg of leflunomide. The AIA rats exhibited body weight loss during the arthritis development, while both JTP-74057 and leflunomide suppressed such weight losses (data not shown). In macroscopic observation, no sign of adverse events was detected with 0.1 mg/kg of JTP-74057 or 10 mg/kg of leflunomide; in particular there was no significant change in liver damage markers (AST, ALT) or kidney damage marker (creatinine) in the rats treated with 0.1 mg/kg of JTP-74057 (data not shown). The maximum tolerated dose (MTD) of JTP-74057 was confirmed as 0.3 mg/kg in rats (data not shown). In the radiographic evaluation of hind paws, bone erosion and destruction were detected in the rats affected with AIA on day 21, and were especially prominent on tarsal and ankle bones (Fig. 4b). JTP-74057 prevented the bone erosions and destructions of hind paws (Fig. 4c), suggesting that MEK inhibitor exerts both an anti-inflammatory effect and a bone-protective effect in rats with AIA.

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JTP-74057/Trametinib administration ameliorates paw swelling in mouse collagen-induced arthritis model [2]
To further compare the pharmacological effect of Trametinib/JTP-74057 with that of leflunomide, we tested these compounds in another widely used RA model, the mouse collagen-induced arthritis model. CII, emulsified with Freund’s complete adjuvant, was injected intradermally on the tail base of DBA1/J mice on day 0 and 21. The paw swelling was periodically scored after the second immunization. JTP-74057 or leflunomide was orally administered once a day from day 21 to day 35. As shown in Fig. 5a, JTP-74057 suppressed development of arthritis in a dose-dependent manner, and the deterioration in clinical score was completely suppressed with 0.3 mg/kg of JTP-74057. Leflunomide also suppressed arthritis development, but did not inhibit it completely, even at 10 mg/kg (Fig. 5b). It was reported that the MTD of JTP-74057 was 3 mg/kg in mice. Consistent with this, there was no sign of adverse effects such as AST elevation or body weight loss in the group treated with 0.3 mg/kg of JTP-74057 (data not shown).

The active form of B-Raf/c-Raf, unphosphorylated MEK1/MEK2, and ERERK2, as well as non-phosphorylated myelin basic protein (MBP), are combined with MOPS buffer containing 12.5 mM MgCl2 and 10 μM ATP in the presence of varying concentrations of GSK1120212. The anti-phospho-MBP antibody can spot MBP that has been phosphorylated.

In 96-well tissue culture plates, exponentially growing cells are precultured for 24 hours before being exposed to Trametinib/GSK1120212. An in vitro toxicology assay kit based on sulforhodamine B measures cell growth. Both adherent and floating cells are collected for the apoptosis assay and fixed with 70% ethanol. The cells are then washed with PBS, suspended in 100 μg/mL RNase and 25 μg/mL propidium iodide (PI), and heated to 37 °C for 30 minutes while kept in the dark. The Cytomics FC500 or Guava EasyCyte plus flow cytometer is used to measure the DNA content of each individual cell.

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PBMCs were cultured in RPMI1640 supplemented with 10% heat-inactivated fetal bovine serum (HI-FBS), and then activated with LPS (human, 1 μg/ml; mouse and rat, 10 μg/ml) in the presence or absence of various concentrations of Trametinib/JTP-74057. For Western blot analyses, cells were lysed at 30 min after stimulation, and the phosphorylation of ERK1/2 was analysed by Western blotting as described previously. For analyses of cytokine production, cell supernatants were collected after overnight stimulation, and concentrations of TNF-α and IL-6 were determined by ELISA. [2]

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Affymetrix expression analyses [3]
16R6-4 was selected for comparison with A375 after compound treatment with GSK2118436 and Trametinib/GSK1120212 alone and in combination with each other for 24 hours. Data analysis was done as described in the Supplementary Methods. The microarray data were deposited in NCBI's Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) and are accessible through GEO Series accession number GSE35230.

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Cell Growth Assay [4]
Cells (2.5 E4) were plated in 96-well plates and treated the following day with increasing concentrations of drug or equamolar dimethyl sulfoxide (DMSO) in triplicate. After 72 hours, redox-dye conversion for each treatment was determined using the Cell Titer-Blue Assay relative to DMSO alone treatment, by flourometry per the manufacturer's instructions. DMSO vehicle at equimolar concentrations had no significant effect on cell viability in all lines.

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Flow Cytometry [4]
For cell cycle analysis, culture supernatants were collected, and combined with cultured cells removed by brief trypsin treatment. Cell were washed twice with PBS and fixed with 70% Ethanol. Cells were stored overnight at −20°C. Cell were then washed twice with PBS, and reconstituted in RNAse A 100ug/mL and 20 ug/mL Propidium Iodide, and stored at 4°C prior to analysis. For apoptosis measurements, cells were similarly collected and washed cells were stained with Annexin V, washed once and re-suspended in 20 ug/mL Propidium Iodide. Cells were analyzed on a FACs Canto and data analyzed using FlowJo. Cell index was determined as the percent of cells in S/G2/M phase normalized to untreated baseline cultures. Means of triplicate experiments were compared with repeated measures one-way ANOVA and Bonferroni's Multiple Comparison Test using Graphpad Prism. Significance indicates a p-value of less than 0.05 given a Confidence Interval of 95% of difference.

Mice: The mice used are BALB/c-nu/nu females. HT-29 cells or COLO205 cells suspended in ice-cold HBSS (-) are subcutaneously injected into the right flank of the mice on day 0 at a density of 5×106 cells/100 µL/site or 1×106 cells per 100 µL, respectively. When the mean tumor volume reaches 100 mm3, the acetic acid-solvated form of Trametinib (JTP-74057, 0.3 mg/kg, or 1 mg/kg) is dissolved in 10% Cremophor EL-10% PEG400 and given orally once daily for 14 days. Two weeks after the start of dosing, the tumor's length [L(mm)] and width [W(mm)] are measured using a microgauge, and the tumor's volume is calculated using the formula tumor volume (mm3)=L×W×W/2.

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Rat adjuvant-induced arthritis [2]
Arthritis was induced by intradermal injection of 0.5 mg of Mycobacterium tuberculosis in 100 μl of paraffin oil into the tail base of male Lewis rats aged 6 weeks (day 0). Normal, untreated rats were used as a control group. Trametinib DMSO solvate and leflunomide were ground and suspended in 0.5% methylcellulose to a volume of 5 ml/kg. On day 0, rats were randomized into 6 groups (6 rats per group) based on body weight. Test drugs were given orally once daily from day 0 to day 21. After arthritis induction, hind paw volume was measured on days 6, 13, 16, and 21 by a water displacement method, using a plethysmometer for rats. Radiographs of both hindlimbs were taken using an X-ray unit on day 21.

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Collagen-induced arthritis [2]
Bovine type II collagen (CII) was dissolved in 0.01 M acetic acid at a concentration of 2 mg/ml and then emulsified in an equal volume of Freund’s complete adjuvant H37Ra. Six-week-old male DBA/1J mice were immunized with 100 μl of the CII emulsion by intradermal injection at the tail base. After 21 days, the mice were randomized into 7 groups (16 mice per group) based on body weight. The mice received the same amount of the CII emulsion to induce arthritis. Trametinib acetic acid solvate was dissolved in 10% Cremophor EL/10% polyethylene glycol 400 solution to a volume of 10 ml/kg. Leflunomide was ground and suspended in 0.5% MC to a volume of 10 ml/kg. Test drug or vehicle was given orally once daily from day 21 to day 35. The clinical score of arthritis was obtained by summing the visual severity grade of each limb, in which swelling of digit and entire paw was scored as follows (maximum score for each limb was 4): for the swelling of digits (0, no swelling; 1, one swollen digit; 2, two or more swollen digits), for the swelling of entire paw (0, no swelling; 1, mild swelling; 2, severe swelling of the entire paw). The score was obtained in a blind manner. Arthritis scores of individual mice were represented as an average score of 4 limbs. CII-specific antibody in sera was measured by a sandwich ELISA method on day 35. Five hundred nanograms of CII were dissolved in 100 μl of PBS, added into 96-well EIA plates, and incubated at 4°C overnight. After washing the excess CII, the plates were blocked with Block Ace for 1 h. Serum from the CIA mice was diluted and added into the plates. After 2 h incubation at RT, the plates were washed, and then the CII-specific antibodies were detected with peroxidase-labeled anti-mouse immunoglobulin (Ig) G antibody.

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Proliferation of lymph node cells from CIA mice [2]
Inguinal lymph node (LN) cells were collected from non-drug-treated CIA mice at 5 days after the second immunization and cultured in a 96-well culture plate at a concentration of 5 × 105 cells/well in RPMI1640 medium containing penicillin–streptomycin, 2-mercaptoethanol, and 10% HI-FBS. CII solution was added to the cells at a final concentration of 10 μg/ml in the presence or absence of test compound, Trametinib or A77 1726. After incubation at 37°C, 5% CO2 for 42 h, 0.5 μCi of [3H]thymidine was added to each well and cultured for 6 h. Incorporated radioactivity was measured using a TopCount microplate scintillation counter.

Absorption, Distribution and Excretion
Trametinib is rapidly and readily absorbed after oral administration. This study investigated the absorption of trametinib in patients with solid tumors and BRAF V600 mutation-positive metastatic melanoma. Following daily administration of 0.125 mg (0.0625 times the recommended adult dose) to 4 mg (twice the recommended adult dose) of trametinib tablets, both Cmax and AUC increased proportionally to the dose. The inter-individual variability in AUC and Cmax at steady state was 22% and 28%, respectively. Repeated daily dosing resulted in trametinib accumulation, with a mean accumulation ratio of 6.0 at a once-daily dose of 2 mg. Steady state was reached on day 15. The mean absolute bioavailability of the oral tablets was 72%, and that of the oral solution was 81%. Tmax was 1.5 hours. Compared to the fasting state, a high-fat, high-calorie meal (approximately 1000 calories) reduced trametinib's AUC by 24%, Cmax by 70%, and delayed Tmax by approximately 4 hours. Following oral administration of [¹⁴C]-trametinib, over 80% of the radioactive material is excreted in feces, while less than 20% is excreted in urine, with the parent molecule accounting for less than 0.1% of the excreted dose. The apparent volume of distribution (V_c/F) is 214 L. The apparent clearance is 4.9 L/h.

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Metabolism/Metabolites
Trametinib is primarily metabolized via deacetylation mediated by carboxylesterases (e.g., carboxylesterase 1b/c and 1b/c) and other hydrolases. Deacetylated metabolites may undergo further glucuronidation. In vitro studies have shown that deacetylation may be accompanied by monooxygenation, hydroxylation, and glucuronidation. CYP3A4-mediated oxidation is a minor pathway. Four metabolites (M1/2/3/4) have been identified in patients with advanced cancer. In vitro studies have shown that the phosphorylation MEK1 inhibitory activity of the M1 and M3 metabolites is approximately the same as or 10-fold lower than that of the parent compound. After a single injection of [¹⁴C]-trametinib, approximately 50% of the circulating radioactivity is the parent compound. Based on metabolite analysis after repeated administration of trametinib, unmetabolized parent drug accounts for 75% or more of the drug-related substances in plasma.

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Biological half-life
The estimated elimination half-life is 3.9 to 4.8 days.

Hepatotoxicity
In large clinical trials, abnormalities in routine liver function tests are common. Among patients treated with trametinib, 39% to 60% experienced elevated serum transaminases, and 24% to 67% experienced elevated alkaline phosphatase. However, ALT elevations exceeding five times the upper limit of normal are uncommon, occurring in only 0% to 5% of cases, and usually resolve rapidly with temporary discontinuation of the drug or dose adjustment. In premarketing controlled trials of trametinib with or without dabrafenib, no clinically significant cases of acute liver injury or liver failure have been reported. There are currently no published reports of clinically significant hepatotoxicity attributable to trametinib. However, it has only been used for a short period. Probability score: E (Unproven but suspected cause of clinically significant liver injury). Pregnancy and Lactation Effects ◉ Overview of Use During Lactation There is currently no information regarding the clinical use of trametinib during lactation. Because trametinib binds to plasma proteins at a rate as high as 97%, its concentration in breast milk may be low. However, its half-life is 3.9 to 4.8 days, and it may accumulate in the infant. The manufacturer recommends discontinuing breastfeeding during trametinib treatment and for 4 months after the last dose.
◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on lactation and breast milk
No published information found as of the revision date.

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Protein binding
Trametinib binds to human plasma proteins at a rate of 97.4%.

[1]. Int J Oncol . 2011 Jul;39(1):23-31.

[2]. Inflamm Res . 2012 May;61(5):445-54.

[3]. Mol Cancer Ther . 2012 Apr;11(4):909-20.

[4]. Clin Cancer Res . 2012 Aug 15;18(16):4345-55.

Pharmacodynamics
Trametinib inhibits the growth of various BRAF V600 mutant positive tumor cells both in vitro and in vivo. Trametinib is often used in combination with the BRAF inhibitor dabrafenib. In BRAF-mutant colorectal cancer, EGFR-mediated MAPK pathway reactivation has been identified as an intrinsic resistance mechanism to BRAF inhibitors. The MAPK pathway is one of the most important pathways in the development of novel anticancer drugs. We performed high-throughput screening on compounds that induce p15INK4b expression and identified JTP-74057 (GSK1120212), which is currently undergoing phase I, II, and III clinical trials. We characterized its in vitro and in vivo antitumor activity. JTP-74057 potently inhibits MEK1/2 kinase activity but has no inhibitory effect on the activity of 98 other kinases. Treatment with JTP-74057 inhibited the growth of most tested colorectal cancer cell lines, accompanied by upregulation of p15INK4b and/or p27KIP1. Daily oral administration of JTP-74057 for 14 days inhibited the growth of HT-29 and COLO205 xenografts in nude mice. Notably, tumor regression was observed only in COLO205 xenografts, and COLO205 cells were more sensitive to JTP-74057-induced apoptosis in vitro than HT-29 cells. Akt inhibitors enhanced JTP-74057-induced apoptosis in HT-29 cells. Finally, JTP-74057 showed additive or synergistic effects when used in combination with standard treatments such as 5-fluorouracil, oxaliplatin, or SN-38. JTP-74057 is a highly specific and potent MEK1/2 inhibitor that exhibits good antitumor activity both in vitro and in vivo. Sensitivity to JTP-74057-induced apoptosis may be an important factor in assessing its in vivo efficacy, and Akt inhibitors can enhance this sensitivity. These results suggest that JTP-74057 has potential value in the treatment of colorectal cancer patients. [1]

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Objective and design: To investigate the effect of the mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 inhibitor JTP-74057 on the development of inflammatory arthritis and to compare its anti-arthritis effect with leflunomide. Materials: Human, mouse and rat peripheral blood mononuclear cells (PBMCs) were used. Lewis rats and DBA/1J mice were used as animal models in this study. Treatment: In in vitro experiments, the concentration range of JTP-74057 was 0.1-100 nM. In in vivo experiments, JTP-74057 (0.01-0.3 mg/kg) and leflunomide (2-10 mg/kg) were administered orally, respectively. Methods: Peripheral blood mononuclear cells (PBMCs) were stimulated with lipopolysaccharide. Adjuvant-induced arthritis (AIA) and type II collagen-induced arthritis (CIA) were induced in Lewis rats and DBA1/J mice, respectively. Results: JTP-74057 inhibited the production of tumor necrosis factor-α and interleukin-6 in PBMCs. 0.1 mg/kg of JTP-74057 or 10 mg/kg of leflunomide almost completely inhibited the development of AIA and CIA. In the CIA model, JTP-74057 (but not leflunomide) inhibited the proliferation of collagen-responsive T cells in vitro, while leflunomide (but not JTP-74057) inhibited the production of anti-collagen antibodies. Conclusion: JTP-74057 has a potent anti-arthritis effect, with a mechanism of action different from leflunomide, suggesting that JTP-74057 may be a novel therapeutic agent for the treatment of rheumatoid arthritis. [2]

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Recent clinical trial results for the BRAF inhibitors GSK2118436 (dabrafenib) and PLX4032 (vemurafenib) have shown encouraging response rates; however, the duration of response is limited. To identify the determinants of acquired resistance to GSK2118436 and strategies to overcome resistance, we isolated GSK2118436-resistant clones from the A375 BRAF (V600E) and YUSIT1 BRAF (V600K) melanoma cell lines. These clones also showed reduced sensitivity to the allosteric mitogen-activated protein/extracellular signal-regulated kinase (MEK) inhibitor GSK1120212 (trametinib). Genetic characterization of these clones revealed that, in the BRAF(V600E) background, there were in-frame deletions of MEK1 (MEK1(K59del)) or NRAS mutations (NRAS(Q61K) and/or NRAS(A146T)), with or without MEK1 (P387S); in the BRAF(V600K) background, NRAS(Q61K) was present. Stable knockdown of NRAS using short hairpin RNA partially restored the sensitivity of mutant NRAS clones to GSK2118436, while expression of NRAS(Q61K) or NRAS(A146T) in A375 parental cells decreased their sensitivity to GSK2118436. Similarly, expression of MEK1(K59del) (but not MEK1(P387S)) decreased the sensitivity of A375 cells to GSK2118436. The combined use of GSK2118436 and GSK1120212 effectively inhibited cell growth in drug-resistant clones, reduced ERK phosphorylation levels, decreased cyclin D1 expression, and increased p27(kip1) protein expression. Furthermore, the combination of GSK2118436 or GSK1120212 with the phosphatidylinositol 3-kinase/mTOR inhibitor GSK2126458 enhanced cell growth inhibition and reduced S6 ribosomal protein phosphorylation levels in these clones. Our results indicate that NRAS and/or MEK mutations are pathogenic factors for BRAF inhibitor resistance in vitro, and that the combination of GSK2118436 and GSK1120212 can overcome this resistance. Moreover, these drug-resistant clones also responded to the combination of GSK2126458 with GSK2118436 or GSK1120212. Clinical trials are currently underway or planned to test these combination therapies. [3]

C26H23FIN5O4

615.39

615.077

C, 50.74; H, 3.77; F, 3.09; I, 20.62; N, 11.38; O, 10.40

871700-17-3

Trametinib (DMSO solvate);1187431-43-1;Trametinib-d4;Trametinib-13C6;Trametinib-13C,d3;2712126-59-3

11707110

white solid powder

1.7±0.1 g/cm3

1.734

2.68

2

6

5

37

1090

0

O=C(C)NC1C=C(N2C3C(=C(N(C)C(C=3C)=O)NC3C(F)=CC(I)=CC=3)C(=O)N(C3CC3)C2=O)C=CC=1

LIRYPHYGHXZJBZ-UHFFFAOYSA-N

InChI=1S/C26H23FIN5O4/c1-13-22-21(23(31(3)24(13)35)30-20-10-7-15(28)11-19(20)27)25(36)33(17-8-9-17)26(37)32(22)18-6-4-5-16(12-18)29-14(2)34/h4-7,10-12,17,30H,8-9H2,1-3H3,(H,29,34)

N-[3-[3-cyclopropyl-5-(2-fluoro-4-iodoanilino)-6,8-dimethyl-2,4,7-trioxopyrido[4,3-d]pyrimidin-1-yl]phenyl]acetamide

JTP-74057; GSK 1120212; GSK1120212; GSK-1120212; JTP74057; Trametinib. Trade name: Mekinist

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: ≥ 2.5 mg/mL (4.06 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 25.0 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (4.06 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: 4% DMSO+corn oil: 3mg/mL

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Solubility in Formulation 4: 6.67 mg/mL (10.84 mM) in 0.5%HPMC 1%Tween80 (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.

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