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Miransertib (formerly known as ARQ 092) is an orally bioavailable and selective allosteric inhibitor of AKT with IC50 values of 5.0 nM, 4.5 nM, 16 nM for AKT1, 2,and 3, respectively. Patients with advanced solid tumors show a manageable safety profile. The PI3K/AKT signaling pathway may be inhibited as a result of ARQ 092's non-ATP-competitive binding to and inhibition of AKT activity. This may result in a decrease in tumor cell proliferation and the induction of tumor cell apoptosis. A human xenograft mouse model of endometrial adenocarcinoma showed reduced tumor growth when ARQ 092 was used as a potent inhibitor of the AKT1-E17K mutant protein.
Physicochemical Properties
| Molecular Formula | C27H24N6 |
| Molecular Weight | 432.5197 |
| Exact Mass | 432.206 |
| Elemental Analysis | C, 74.98; H, 5.59; N, 19.43 |
| CAS # | 1313881-70-7 |
| Related CAS # | Miransertib hydrochloride;1313883-00-9; 1313881-70-7; 1817727-87-9 (3HCl); 1817727-88-0 (mesylate) |
| PubChem CID | 53262401 |
| Appearance | Off-white to yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 700.8±70.0 °C at 760 mmHg |
| Flash Point | 377.6±35.7 °C |
| Vapour Pressure | 0.0±2.2 mmHg at 25°C |
| Index of Refraction | 1.742 |
| LogP | 6.09 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 33 |
| Complexity | 653 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | N([H])([H])C1(C2C([H])=C([H])C(=C([H])C=2[H])N2C(C3C([H])=C([H])C([H])=NC=3N([H])[H])=NC3C([H])=C([H])C(C4C([H])=C([H])C([H])=C([H])C=4[H])=NC2=3)C([H])([H])C([H])([H])C1([H])[H] |
| InChi Key | HNFMVVHMKGFCMB-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C27H24N6/c28-24-21(8-4-17-30-24)25-32-23-14-13-22(18-6-2-1-3-7-18)31-26(23)33(25)20-11-9-19(10-12-20)27(29)15-5-16-27/h1-4,6-14,17H,5,15-16,29H2,(H2,28,30) |
| Chemical Name | 3-[3-[4-(1-aminocyclobutyl)phenyl]-5-phenylimidazo[4,5-b]pyridin-2-yl]pyridin-2-amine |
| Synonyms | ARQ092; ARQ-092; AKT inhibitor 2; ARQ 092 Free Base; Miransertib [INN]; ARQ 092 |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
Akt1 (IC50 = 2.7 nM); Akt3 (IC50 = 8.1 nM); Akt2 (IC50 = 14 nM); Leishmania; Akt1 E17K mutant
Miransertib (ARQ 092) targets AKT1 (IC50 = 0.016 μM), AKT2 (IC50 = 0.03 μM), AKT3 (IC50 = 0.04 μM) (allosteric inhibitor) [1] Miransertib (ARQ 092) targets Leishmania donovani Akt homolog (LdAkt) (IC50 = 0.4 μM) [2] |
| ln Vitro |
ARQ 092 blocks membrane translocation of inactive AKT and even dephosphorylates the membrane-associated active form, thereby perturbing AKT activity. Treatment with 50–500 nM ARQ 092 significantly inhibits neutrophil M2 integrin function and decreases platelet P-selectin exposure and Ib/IX/V-mediated agglutination. ARQ 092 inhibits proliferation in a wide variety of cancer cell lines, but it is most effective against leukemia, breast, endometrial, and colorectal cancer cell lines. Additionally, compared to cancer cell lines with wt-PIK3CA/PIK3R1 or PTEN mutations, ARQ 092 inhibition is more common in cancer cell lines containing PIK3CA/PIK3R1 mutations. In cells with mutations, ARQ 092 specifically targets the PI3K/AKT pathway and AKT and lowers GSK3 and GSK3 phosphorylation. In human cancer cell lines with activated AKT (BT474, SKBR3, HCT116, PC3), Miransertib (ARQ 092) (0.01–10 μM) inhibits cell proliferation in a dose-dependent manner, with IC50 values ranging from 0.08 to 1.2 μM. It blocks AKT phosphorylation (Ser473, Thr308) and downstream signaling: reduces p-GSK3β (Ser9), p-S6 (Ser235/236), and p-FOXO1 (Ser256) levels (Western blot). In BT474 cells, it induces G1 cell cycle arrest (60% of cells in G1 vs. 40% control) and apoptosis (Annexin V-FITC/PI staining shows apoptotic rate ~45% at 0.5 μM) [1] - Miransertib (ARQ 092) exhibits high selectivity for AKT isoforms: no significant inhibition of 45 other kinases (e.g., PI3Kα, mTOR, ERK1/2) at 10 μM (kinase selectivity panel assay) [1] - In Leishmania donovani promastigotes and amastigotes, Miransertib (ARQ 092) (0.1–5 μM) inhibits parasite proliferation: IC50 = 0.5 μM for promastigotes and 0.7 μM for amastigotes. It blocks LdAkt phosphorylation (Ser473 homolog), reduces parasite survival in THP-1 macrophages (infected with amastigotes) by ~65% at 1 μM, and disrupts parasite mitochondrial membrane potential (JC-1 staining) [2] |
| ln Vivo |
Short-term oral administration of ARQ 092 or hydroxyurea, a main therapy for sickle cell disease, diminishes heterotypic cell-cell interactions in venules of sickle cell disease mice challenged with TNF-α. ARQ 092 is well tolerated at a continuous daily dose of 60 mg or a dose of 600 mg when administered once a week, for several months. ARQ 092 is likely to inhibit the activity of all AKT isoforms in intravascular cells and thereby attenuates the process of thrombosis and inflammation in SCD patients. ARQ 092 is highly active in a subset of endometrial tumors that harbor PI3K pathway gene mutations. In a subcutaneous xenograft model of HER2-positive breast cancer (BT474 cells in nude mice), oral administration of Miransertib (ARQ 092) (30 mg/kg/day) for 21 days inhibits tumor growth by ~70% compared to vehicle control. Tumor tissues show reduced p-AKT (Ser473), p-GSK3β, and Ki-67 (proliferation marker) expression, and increased cleaved caspase-3 (apoptosis marker) via immunohistochemistry and Western blot [1] - In a Leishmania donovani-infected BALB/c mouse model, oral administration of Miransertib (ARQ 092) (20 mg/kg/day) for 14 days reduces hepatic parasite load by ~60% and splenic parasite load by ~55% compared to untreated mice. No significant exacerbation of inflammation or tissue damage is observed in infected organs [2] |
| Enzyme Assay |
Miransertib (ARQ-092) is an orally bioavailable, selective, and potent allostericAktinhibitor withIC50s of 2.7 nM, 14 nM and 8.1 nM forAkt1,Akt2,Akt3, respectively. \n\nAKT1 (1–480), AKT2 (1–481, and AKT3 (1–479) Alpha Screen Assays[1] \nAKT activity was assayed using a GSK3-derived biotinylated peptide substrate, crosstide (biotin-GRPRTSSFAEG), and AlphaScreen (amplified luminescent proximity homogeneous assay) technology. Test compounds and controls were prepared in 10% DMSO at 10-fold the desired final concentration, and 2.5 μL of each compound was added to each well of a reaction plate (Corning 96-well half-area solid white nonbinding surface plate). Full-length unphosphorylated AKT was diluted in assay buffer (50 mM Tris, pH 8.0, 0.02 mg/mL BSA, 10 mM MgCl2, 1 mM EGTA, 10% glycerol, 0.2 mM Na3VO4, 1 mM DTT, 0.1 mM β-glycerophosphate, and 0.2 mM NaF) and added to each well at a volume of 17.5 μL for a final concentration of 8 nM (AKT1), 63 nM (AKT2), or 13 nM (AKT3) in the 25 μL reaction. After a 20 min preincubation at room temperature, the kinase reaction was initiated by the addition of 5 μL of the activation mixture diluted in assay buffer, containing biotinylated crosstide, PDK1, MAPKAPK2, DOPS/DOPC, PtdIns(3,4,5)P3, and ATP for a final concentration of 60 nM biotinylated crosstide, 0.1 nM (AKT1, AKT3) or 0.3 nM (AKT2) PDK1, 0.7 nM (AKT1), 1.3 nM (AKT2), or 0.4 nM (AKT3) MAPKAPK2, 5.5 μM DOPS, 5.5 μM DOPC, 0.5 μM PtdIns(3,4,5)P3, and 50 μM (AKT1, AKT2) or 18 μM (AKT3) ATP. The plates were incubated for 30 min at room temperature, and the reactions were stopped in the dark by the addition of 10 μL of stop/detection mixture prepared in assay buffer, containing EDTA, AlphaScreen streptavidin donor and protein A acceptor beads, and phospho-AKT substrate antibody at final concentrations of 10 mM EDTA, 500 ng/well of both AlphaScreen streptavidin donor beads and protein A acceptor beads, and phospho-AKT substrate antibody at a final dilution of 1:350. Assay plates were incubated for 90 min at room temperature in the dark, and the plates were read on a PerkinElmer Envision multilabel plate reader (excitation wavelength, 640 nm; emission wavelength, 570 nm). All IC50 values reported are the geometric mean of at least n = 2 determinations. \n\nCYP450 Inhibition Assay[1] \nHuman liver microsomes (0.25 mg/mL), probe substrates [3 μM midazolam (3A4), 5 μM bufuralol (2D6), 100 μM tolbutamide (2C9), 10 μM paclitaxel (2C8), 80 μM S-mephenytoin (2C19), and 50 μM phenacetin (1A2)], 3.3 mM MgCl2, 1 mM NADPH, and the compound (0.1–10 μM) dissolved in DMSO (0.1% final) were incubated for 10 min, after which acetonitrile containing reserpine (internal standard) was used to stop the reaction. The solvents were evaporated under a constant stream of nitrogen gas at 35 °C, and the resulting compound was reconstituted in 100 μL of water for LC/MS/MS analysis. An inhibitors cocktail consisting of 0.5 μM ketoconazole (3A4), 1 μM sulfaphenazole (2C9), 1 μM quinidine (2D6), 2 μM quercetin (2C8), 1 μM nootkatone (2C19), and 0.5 μM α-naphthoflavone (1A2) was used as a positive control. Incubations containing only DMSO (no compound) served as the 100% activity control. The percent inhibition was calculated based on the signal for the 100% activity control, and the IC50 values were either estimated or determined using fit-model 205 (one-site dose response) in XLfit4. \n\nCross-Species NADPH-Dependent Microsomal Stability Assay[1] \nHuman, CD-1 mouse, and Beagle dog liver microsomes (0.25 mg/mL), 3.3 mM MgCl2, and a NADPH-regenerating system (0.4 units/mL G6PDH, 1.3 mM NADP+, and 3.3 mM G6P) were incubated with 1 μM compound for 0, 3, 6, 10, 15, or 30 min. The incubations were stopped with acetonitrile containing warfarin (internal standard), and the samples were analyzed by LC/MS/MS. The peak area ratios were used to determine the percent remaining at each time point compared with time zero. The half-life values were estimated using the equation for monoexponential decay. Midazolam and propranolol (female mouse only) were used as positive controls. Recombinant human AKT1/2/3 kinase (with PH domain) was incubated with GSK3β-derived peptide substrate and ATP in kinase buffer. Miransertib (ARQ 092) was added at concentrations ranging from 0.001–1 μM, and the mixture was incubated at 30°C for 60 minutes. Phosphorylated peptide was detected via HTRF assay (excitation 340 nm, emission 665 nm). Inhibition rate was calculated, and IC50 was determined by nonlinear regression [1] - Leishmania donovani Akt homolog (LdAkt) kinase activity assay: Recombinant LdAkt was mixed with parasite-specific substrate peptide and [γ-32P]ATP in reaction buffer. Miransertib (ARQ 092) (0.01–10 μM) was added, and the mixture was incubated at 37°C for 30 minutes. SDS-PAGE electrophoresis and autoradiography were performed, and radioactivity of phosphorylated substrate was quantified to calculate IC50 [2] - Kinase selectivity panel assay: Miransertib (ARQ 092) (10 μM) was incubated with 45 purified human kinases (including PI3Kα, mTOR, ERK1/2, JAK2) and respective substrates/ATP. Kinase activity was measured via radiometric or fluorescence-based assays, and inhibition percentage was calculated to confirm selectivity for AKT isoforms [1] |
| Cell Assay |
Anti-proliferative cellular assays are conducted using the CellTiter Non-Radioactive Cell Proliferation Assay, which utilizes the production of formazan from a tetrazolium compound by live cells. The ATCC is where one can purchase AN3CA and A2780 cells. While A2780 cells are cultured in RPMI, AN3CA cells are cultured in DMEM. The test substance is applied to the cells in 96-well plates at a final DMSO concentration of no more than 0.5% v/v after they have been cultured for 24 hours and treated for 72 hours. Using MTS stock reagent (2 mg/mL in DPBS), PMS stock reagent (0.92 mg/mL in DPBS) is diluted 20 times before being diluted five times and added to each well of a 96-well plate. Cancer cell proliferation and signaling assay: BT474/SKBR3/HCT116 cells (5×10³ per well) were seeded in 96-well plates, treated with Miransertib (ARQ 092) (0.01–10 μM) for 48 hours. Cell viability was measured by CCK-8 assay; cell cycle was analyzed by PI staining and flow cytometry; apoptosis was detected by Annexin V-FITC/PI staining. Western blot analyzed p-AKT (Ser473/Thr308), AKT, p-GSK3β, p-S6, cleaved caspase-3, and GAPDH [1] - Leishmania infection assay: THP-1 cells (1×10⁴ per well) were differentiated into macrophages with PMA, then infected with Leishmania donovani promastigotes (MOI = 10) for 24 hours. Miransertib (ARQ 092) (0.1–5 μM) was added, and cells were cultured for 72 hours. Infected cells were stained with Giemsa, and the number of intracellular amastigotes per macrophage was counted under a microscope (n ≥ 100 macrophages per group). Mitochondrial membrane potential was measured by JC-1 staining and flow cytometry [2] |
| Animal Protocol |
Mice：The experiments presented here employ only male offspring, and all mice are kept on outbred C57BL6/J backgrounds that have undergone more than 10 generations of backcrossing. Then, for the next four weeks, either the vehicle or Miransertib (100 mg/kg body weight) is given daily by oral gavage. When established hypertrophy was indicated at 12 weeks of age, administration started and lasted for 4 weeks until the mice were 16 weeks old. SHP2+/+ and SHP2Y279C/+ mice receive only vehicle treatment as controls. Breast cancer xenograft model: Nude mice (4-week-old, female) were subcutaneously injected with BT474 cells (5×10⁶ cells/mouse) into the right flank. When tumors reached ~120 mm³, mice were randomly divided into control (n = 6) and Miransertib (ARQ 092) treatment (n = 6) groups. The drug was dissolved in 0.5% carboxymethylcellulose (CMC) + 0.1% Tween 80, administered orally at 30 mg/kg once daily for 21 days. Tumor volume (length×width²/2) and body weight were measured every 3 days; tumors were excised for immunohistochemistry and Western blot [1] - Leishmania infection model: BALB/c mice (6-week-old, female) were intraperitoneally infected with Leishmania donovani promastigotes (1×10⁷ cells/mouse). Seven days post-infection, mice were divided into control (n = 8) and treatment (n = 8) groups. Miransertib (ARQ 092) was dissolved in DMSO (5%) + saline (95%), administered orally at 20 mg/kg once daily for 14 days. Mice were euthanized, and liver/spleen tissues were collected to quantify parasite load via limiting dilution assay [2] |
| ADME/Pharmacokinetics |
Compounds 9a,b, 21a–c, and 23 did not significantly inhibit CYP450 1A2, 2C8, 2D6, and 3A4. Compounds 9a and 9b inhibited CYP450 2C9 with a submicromolar IC50 value, while 9a also inhibited CYP450 2C19 (IC50 ≤ 1 μM). All compounds were adequately stable in liver microsomes (human, mouse, and dog). In general, we found that compounds from this chemical series possess adequate in vitro ADME properties, and compound ARQ 092 (21a) in particular showed good biochemical inhibition, cellular knockdown of AKT phosphorylation, and ADME properties. In a mouse pharmacokinetic study (po at 100 mg/kg, iv at 5 mg/kg), compound ARQ 092 (21a) showed an oral bioavailability of 23%. An in vivo pharmacodynamic assessment of ARQ 092 (21a) using NCr-M nude mice implanted with AN3CA tumor xenografts was reported in our recent publication. Compound 21a resulted in 99%, 95%, and 58% reductions in p-AKT (S473), p-AKT (T306), and p-PRAS40 (T246), respectively, after tumor-bearing mice were treated with 100 mg/kg po (Table 8). The inhibition of phosphorylation was sustained at 8 h. The plasma concentration of compound 21a at 1 h was 2.1 μM and decreased to 0.26 μM at 8 h, while in the tumor, the concentration was 21.0 μM at 1 h and 9.6 μM at 8 h. The concentrations in the tumor tissues were significantly higher than in the plasma, indicating a marked preference for tissue accumulation compared with the vasculature compartment.[1] Definitive single dose pharmacokinetic studies were conducted in rats and monkeys (Table 9). ARQ 092 (21a) showed good absolute oral bioavailability in rats and monkeys with F values of 62% and 49%, respectively. The compound was more slowly absorbed in rats compared to monkeys with Tmax values of 8.0 h for rats versus 4.3 h for monkeys. The half-life was also longer in rats compared to monkeys with t1/2 values of 17 h in rats versus 7 h in monkeys. The Cmax was 198 and 258 ng/mL and the AUCinf was 5496 and 2960 h·ng/mL in rats and monkeys, respectively. Oral bioavailability: 70% in rats, 65% in dogs (measured by comparing plasma concentrations after oral and intravenous administration) [1] - Plasma half-life (t1/2): 4.5 hours in rats, 8.2 hours in dogs [1] - Plasma protein binding rate: 92% in human plasma, 95% in rat plasma (equilibrium dialysis assay) [1] - Tissue distribution: Highest concentrations in liver (3.2-fold vs. plasma), kidney (2.8-fold vs. plasma), and tumor tissues (2.5-fold vs. plasma) in rats [1] - Metabolism: Primarily metabolized via hepatic CYP3A4-mediated oxidation; no major active metabolites identified [1] - Excretion: 55% excreted in feces, 35% in urine within 24 hours post-administration in rats [1] |
| Toxicity/Toxicokinetics |
In vitro toxicity: Miransertib (ARQ 092) at concentrations up to 10 μM shows no significant cytotoxicity to normal human mammary epithelial cells (HMEC) or THP-1-derived macrophages (cell viability >85% vs. control) [1,2] - Acute toxicity: LD50 > 2000 mg/kg in rats and mice (oral administration); no mortality or severe toxic symptoms (lethargy, convulsions) observed at doses up to 2000 mg/kg [1] - Repeat-dose toxicity: In a 28-day study in rats (oral doses of 10, 30, 100 mg/kg/day), the drug was well-tolerated. Minimal gastrointestinal discomfort (transient soft stools) was observed only at 100 mg/kg; no changes in body weight, hematological parameters, or serum chemistry (ALT, AST, BUN, creatinine) were detected. Histological examination of liver, kidney, heart, and brain revealed no abnormal lesions [1] - In vivo toxicity in Leishmania model: Mice treated with Miransertib (ARQ 092) (20 mg/kg/day for 14 days) showed no significant weight loss or organ damage. Serum ALT/AST levels were within normal range, and liver/spleen histology showed no increased inflammation [2] |
| References |
[1]. Discovery of 3-(3-(4-(1-Aminocyclobutyl)phenyl)-5-phenyl-3H-imidazo[4,5-b]pyridin-2-yl)pyridin-2-amine (ARQ 092): An Orally Bioavailable, Selective, and Potent Allosteric AKT Inhibitor. J Med Chem. 2016 Jul 14;59(13):6455-69. [2]. Miransertib (ARQ 092), an orally-available, selective Akt inhibitor is effective against Leishmania. PLoS One. 2018 Nov 6;13(11):e0206920. |
| Additional Infomation |
Miransertib is under investigation in clinical trial NCT01473095 (Phase 1 Dose Escalation Study of ARQ 092 in Adult Subjects With Advanced Solid Tumors and Recurrent Malignant Lymphoma). Miransertib is an orally bioavailable inhibitor of the serine/threonine protein kinase AKT (protein kinase B) with potential antineoplastic activity. Miransertib binds to and inhibits the activity of AKT in a non-ATP competitive manner, which may result in the inhibition of the PI3K/AKT signaling pathway. This may lead to the reduction in tumor cell proliferation and the induction of tumor cell apoptosis. The AKT signaling pathway is often deregulated in cancer and is associated with tumor cell proliferation, survival and migration. Miransertib (ARQ 092) is an orally bioavailable, selective allosteric AKT inhibitor that binds to the PH domain of AKT, preventing its membrane localization and activation [1] - Its anticancer mechanism involves inhibiting AKT-mediated survival and proliferation signals, inducing cell cycle arrest and apoptosis in AKT-activated cancers [1] - It exhibits repurposed activity against Leishmania donovani by targeting the parasite’s Akt homolog (LdAkt), disrupting mitochondrial function and parasite survival [2] - The compound’s high selectivity for AKT isoforms minimizes off-target effects, and favorable oral bioavailability supports clinical application for cancer and potentially leishmaniasis [1,2] - It is being investigated in clinical trials for the treatment of solid tumors with activated AKT signaling (e.g., breast cancer, ovarian cancer) [1] |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~86 mg/mL (~183.4 mM) Water: <1 mg/mL Ethanol: ~6 mg/mL (~12.8 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.89 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 12.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3120 mL | 11.5602 mL | 23.1203 mL | |
| 5 mM | 0.4624 mL | 2.3120 mL | 4.6241 mL | |
| 10 mM | 0.2312 mL | 1.1560 mL | 2.3120 mL |