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]

---

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.

---

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]

---

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.

---

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.

---

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]

---

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.

---

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.

---

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.

---

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.

---

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.

---

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
Following oral administration, trametinib is rapidly and readily absorbed. The absorption was examined in patients with solid tumours and BRAF V600 mutation-positive metastatic melanoma. Following the administration of trametinib tablets 0.125 mg (0.0625 times the approved recommended adult dosage) to 4 mg (2 times the approved recommended adult dosage) daily, both Cmax and AUC increased dose-proportionally. Intersubject variability in AUC and Cmax at steady state is 22% and 28%, respectively. Trametinib accumulates with daily repeat dosing with a mean accumulation ratio of 6.0 at 2 mg once daily dose. Steady-state was achieved by Day 15. The mean absolute bioavailability of trametinib is 72% for oral tablets and 81% for oral solution. The Tmax is 1.5 hours. A high-fat, high-calorie meal (approximately 1000 calories) decreased trametinib AUC by 24% and Cmax by 70%, and delayed Tmax by approximately four hours as compared with fasted conditions.
Following oral administration of [¹⁴C]-trametinib, greater than 80% of excreted radioactivity was recovered in the feces while less than 20% of excreted radioactivity was recovered in the urine with less than 0.1% of the excreted dose as the parent molecule.
The apparent volume of distribution (V_c/F) is 214 L.
The apparent clearance is 4.9 L/h.

---

Metabolism / Metabolites
Trametinib predominantly undergoes deacetylation mediated by carboxylesterases (i.e., carboxylesterase 1b/c and 2) and other hydrolytic enzymes. The deacetylated metabolite may further be glucuronidated. _In vitro_ findings suggest that deacetylation may also be accompanied by mono-oxygenation, hydroxylation, and glucuronidation. CYP3A4-mediated oxidation is a minor pathway. Four metabolites (M1/2/3/4) have been characterized in patients with advanced cancers. _In vitro_, the M1 and M3 metabolites demonstrated approximately equal or 10-fold less potent phospho-MEK1-inhibiting activity than the parent compound. Following a single dose of [¹⁴C]-trametinib, approximately 50% of circulating radioactivity represented the parent compound. According to findings from metabolite profiling after repeat dosing of trametinib, unchanged parent drug accounted for greater than or equal to 75% of drug-related material in plasma.

---

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

Hepatotoxicity
In large clinical trials, abnormalities in routine liver tests were common with serum aminotransferase elevations occurring in 39% to 60% and alkaline phosphatase in 24% to 67% of patients treated with trametinib. However, elevations in ALT above 5 times the ULN were uncommon, occurring in 0% to 5% of patients and generally resolving rapidly with temporary discontinuation or dose adjustment. In the prelicensure controlled trials of trametinib with or without dabrafenib, no cases of clinically apparent acute liver injury or hepatic failure were reported. There have yet to be published cases of clinically apparent hepatotoxicity attributed to trametinib. However, it has been used for a short time only.
Likelihood score: E* (unproven but suspected cause of clinically apparent liver injury).

---

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of trametinib during breastfeeding. Because trametinib is 97% bound to plasma proteins, the amount in milk is likely to be low. However, its half-life is 3.9 to 4.8 days and it might accumulate in the infant. The manufacturer recommends that breastfeeding be discontinued during trametinib therapy and for 4 months after the last dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

---

Protein Binding
Trametinib is 97.4% bound to human plasma proteins.

[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 cell growth of various BRAF V600 mutation-positive tumours _in vitro_ and _in vivo_. Trametinib is often used in combination with dabrafenib, a BRAF inhibitor. In BRAF-mutant colorectal cancer, induction of EGFR-mediated MAPK pathway re-activation has been identified as a mechanism of intrinsic resistance to BRAF inhibitors.

---

The MAPK pathway is one of the most important pathways for novel anticancer drug development. We performed high-throughput screening for compounds that induce expression of p15INK4b, and identified JTP-74057 (GSK1120212), which is being evaluated in ongoing phase I, II and III clinical trials. We characterized its antitumor activities in vitro and in vivo. JTP-74057 strongly inhibited MEK1/2 kinase activities, but did not inhibit another 98 kinase activities. Treatment by JTP-74057 resulted in growth inhibition accompanied with upregulation of p15INK4b and/or p27KIP1 in most of the colorectal cancer cell lines tested. Daily oral administration of JTP-74057 for 14 days suppressed tumor growth of HT-29 and COLO205 xenografts in nude mice. Notably, tumor regression was observed only in COLO205 xenografts, and COLO205 was much more sensitive to JTP-74057-induced apoptosis than HT-29 in vitro. Treatment with an Akt inhibitor enhanced the JTP-74057-induced apoptosis in HT-29 cells. Finally, JTP-74057 exhibited an additive or a synergistic effect in combination with the standard-of-care agents, 5-fluorouracil, oxaliplatin or SN-38. JTP-74057, a highly specific and potent MEK1/2 inhibitor, exerts favorable antitumor activities in vitro and in vivo. Sensitivity to JTP-74057-induced apoptosis may be an important factor for the estimation of in vivo efficacy, and sensitivity was enhanced by an Akt inhibitor. These results suggest the usefulness of JTP-74057 in therapeutic applications for colorectal cancer patients.[1]

---

Objective and design: To examine the effects of a mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2-inhibitor, JTP-74057, on inflammatory arthritis development, and compare its anti-arthritic effect with leflunomide. Materials: Human, mouse, and rat peripheral blood mononuclear cells (PBMCs) were used. Lewis rats and DBA/1J mice were used for animal models. Treatment: JTP-74057 was tested between 0.1-100 nM in in-vitro studies. JTP-74057 (0.01-0.3 mg/kg) and leflunomide (2-10 mg/kg) were administered orally in vivo. Methods: PBMCs were stimulated with lipopolysaccharide. Adjuvant-induced arthritis (AIA) and type II collagen-induced arthritis (CIA) was induced in Lewis rats or DBA1/J mice, respectively. Results: JTP-74057 blocked tumor necrosis factor-α and interleukin-6 production from PBMCs. AIA and CIA development were suppressed almost completely by 0.1 mg/kg of JTP-74057 or 10 mg/kg of leflunomide. In the CIA, JTP-74057, but not leflunomide, suppressed collagen-reactive T-cell proliferation ex vivo, whereas leflunomide, but not JTP-74057, suppressed anti-collagen antibody production. Conclusions: JTP-74057 exerts potent anti-arthritic effects with a different profile from leflunomide, suggesting that JTP-74057 may be useful as a new therapeutic reagent in the treatment of rheumatoid arthritis.[2]

---

Recent results from clinical trials with the BRAF inhibitors GSK2118436 (dabrafenib) and PLX4032 (vemurafenib) have shown encouraging response rates; however, the duration of response has been limited. To identify determinants of acquired resistance to GSK2118436 and strategies to overcome the resistance, we isolated GSK2118436 drug-resistant clones from the A375 BRAF(V600E) and the 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 identified an in-frame deletion in MEK1 (MEK1(K59del)) or NRAS mutation (NRAS(Q61K) and/or NRAS(A146T)) with and without MEK1(P387S) in the BRAF(V600E) background and NRAS(Q61K) in the BRAF(V600K) background. Stable knockdown of NRAS with short hairpin RNA partially restored GSK2118436 sensitivity in mutant NRAS clones, whereas expression of NRAS(Q61K) or NRAS(A146T) in the A375 parental cells decreased sensitivity to GSK2118436. Similarly, expression of MEK1(K59del), but not MEK1(P387S), decreased sensitivity of A375 cells to GSK2118436. The combination of GSK2118436 and GSK1120212 effectively inhibited cell growth, decreased ERK phosphorylation, decreased cyclin D1 protein, and increased p27(kip1) protein in the resistant clones. Moreover, the combination of GSK2118436 or GSK1120212 with the phosphoinositide 3-kinase/mTOR inhibitor GSK2126458 enhanced cell growth inhibition and decreased S6 ribosomal protein phosphorylation in these clones. Our results show that NRAS and/or MEK mutations contribute to BRAF inhibitor resistance in vitro, and the combination of GSK2118436 and GSK1120212 overcomes this resistance. In addition, these resistant clones respond to the combination of GSK2126458 with GSK2118436 or GSK1120212. Clinical trials are ongoing or planned to test these combinations.[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.

---

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.

---

View More

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

---

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.

---

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