 Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C28H31F2N7O4S |
| Molecular Weight | 599.652051210403 |
| Exact Mass | 599.212 |
| CAS # | 2241039-81-4 |
| PubChem CID | 135240395 |
| Appearance | White to off-white solid powder |
| LogP | 2.2 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 11 |
| Rotatable Bond Count | 9 |
| Heavy Atom Count | 42 |
| Complexity | 1010 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | S(C)(C1C=CC=C(C=1F)NC1=NC=C(C(C2=CNC3C2=CC=CC=3NC([C@@H](COC)N2CCN(C)CC2)=O)=N1)F)(=O)=O |
| InChi Key | JNUZADQZHYFJGW-JOCHJYFZSA-N |
| InChi Code | InChI=1S/C28H31F2N7O4S/c1-36-10-12-37(13-11-36)22(16-41-2)27(38)33-21-8-4-6-17-18(14-31-26(17)21)25-19(29)15-32-28(35-25)34-20-7-5-9-23(24(20)30)42(3,39)40/h4-9,14-15,22,31H,10-13,16H2,1-3H3,(H,33,38)(H,32,34,35)/t22-/m1/s1 |
| Chemical Name | (2R)-N-[3-[5-fluoro-2-(2-fluoro-3-methylsulfonylanilino)pyrimidin-4-yl]-1H-indol-7-yl]-3-methoxy-2-(4-methylpiperazin-1-yl)propanamide |
| 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 |
Londamocitinib (AZD4604; JAK1-IN-7) targets Janus kinase 1 (JAK1) (IC50 = 0.003 μM for human JAK1 kinase activity; Ki = 0.0015 μM) [2][3] Londamocitinib (AZD4604; JAK1-IN-7) shows high selectivity over other JAK family members (JAK2: IC50 = 0.32 μM; JAK3: IC50 = 0.45 μM; TYK2: IC50 = 0.28 μM; selectivity indices > 90 vs. JAK1) [2][3] Londamocitinib (AZD4604; JAK1-IN-7) exhibits no significant inhibition of 450+ other kinases (IC50 > 10 μM) [2] |
| ln Vitro |
- JAK1 kinase inhibitory activity: Londamocitinib (AZD4604; JAK1-IN-7) potently inhibits recombinant human JAK1 kinase activity in a dose-dependent manner, with IC50 = 0.003 μM and Ki = 0.0015 μM. It competitively binds to the ATP-binding pocket of JAK1 [2][3] - Inhibition of JAK1-mediated signaling: The compound (0.001-0.1 μM) dose-dependently blocks IL-4/IL-13-induced STAT6 phosphorylation and IL-6-induced STAT3 phosphorylation in human peripheral blood mononuclear cells (PBMCs) and bronchial epithelial cells (BECs). At 0.01 μM, STAT6 phosphorylation is reduced by 85% (IL-4-induced) and 82% (IL-13-induced) [2][3] - Suppression of pro-inflammatory mediator production: Londamocitinib (AZD4604; JAK1-IN-7) (0.005-0.1 μM) inhibits IL-4/IL-13-induced eotaxin-1, MUC5AC, and IL-5 production in BECs and lung fibroblasts. At 0.05 μM, eotaxin-1 and MUC5AC levels are reduced by 70% and 65% respectively [2][3] - High kinase selectivity: The compound shows minimal inhibition of JAK2, JAK3, and TYK2 (IC50 > 0.28 μM) and no significant activity against 450+ other kinases (e.g., EGFR, ERK, CDK2) at 10 μM [2] - Minimal cytotoxicity: At concentrations up to 20 μM, Londamocitinib (AZD4604; JAK1-IN-7) exhibits no obvious cytotoxicity to BECs, PBMCs, or normal human lung fibroblasts (cell viability > 90%) [2][3] |
| ln Vivo |
- Efficacy in OVA-induced allergic asthma mouse model: BALB/c mice with OVA-induced asthma were administered Londamocitinib (AZD4604; JAK1-IN-7) via inhalation (0.1 mg/kg, 0.3 mg/kg, 1 mg/kg) once daily for 7 days. The 1 mg/kg dose reduces airway eosinophilic infiltration by 80%, peribronchial inflammation by 75%, and BAL fluid IL-4/IL-5/IL-13 levels by 70-78% compared to vehicle control. It also improves airway hyperresponsiveness (AHR) to methacholine (PC20 increased by 2.3-fold) [2][3] - Lung-targeted distribution: Inhaled Londamocitinib (AZD4604; JAK1-IN-7) (1 mg/kg) achieves lung tissue concentration of 12 μM at 1 hour post-dose, with minimal systemic exposure (plasma concentration = 0.08 μM), resulting in a lung/plasma ratio of 150 [2] - Mechanism validation in vivo: Lung tissues from treated mice (1 mg/kg) show reduced JAK1 phosphorylation (72% reduction) and STAT6 phosphorylation (68% reduction) compared to control, confirming JAK1 pathway inhibition [2][3] - Tolerability: No significant body weight loss (< 5%) or obvious toxic signs (lethargy, respiratory distress) are observed in treated mice. Serum ALT, AST, creatinine, and urea nitrogen levels remain within normal ranges [2][3] |
| Enzyme Assay |
- JAK1 kinase activity assay: Recombinant human JAK1 catalytic domain was mixed with ATP (10 μM), peptide substrate (derived from STAT3), and gradient concentrations of Londamocitinib (AZD4604; JAK1-IN-7) (0.0001-1 μM) in kinase buffer (pH 7.5). The mixture was incubated at 30°C for 1 hour, and phosphorylated substrate was detected by HTRF assay. IC50 was calculated by plotting inhibition rate against drug concentration [2][3] - Surface plasmon resonance (SPR) binding assay: Recombinant human JAK1 catalytic domain was immobilized on a sensor chip. Londamocitinib (AZD4604; JAK1-IN-7) at gradient concentrations (0.001-10 μM) was injected, and binding affinity was measured at 25°C. The equilibrium dissociation constant (KD) was 0.0012 μM, confirming high-affinity binding to JAK1 [2] - Kinase selectivity panel assay: Londamocitinib (AZD4604; JAK1-IN-7) (10 μM) was screened against a panel of 456 human kinases. Inhibition of each kinase was measured by radiometric assay or fluorescence-based assay to evaluate off-target activity [2] |
| Cell Assay |
- Cytokine-induced STAT phosphorylation assay: Human BECs or PBMCs were seeded into 6-well plates (5×10⁵ cells/well) and treated with Londamocitinib (AZD4604; JAK1-IN-7) (0.001-0.1 μM) for 1 hour, then stimulated with IL-4 (10 ng/mL) or IL-13 (10 ng/mL) for 30 minutes. Cells were lysed, and p-STAT6, total STAT6, p-JAK1, and GAPDH were detected by western blot. Band intensities were quantified by densitometry [2][3] - Pro-inflammatory mediator production assay: Human lung fibroblasts were seeded into 96-well plates (1×10⁴ cells/well) and pre-treated with Londamocitinib (AZD4604; JAK1-IN-7) (0.005-0.1 μM) for 1 hour, then stimulated with IL-4/IL-13 (10 ng/mL each) for 24 hours. Culture supernatants were collected, and eotaxin-1, MUC5AC, and IL-5 levels were measured by ELISA [2][3] - Cell viability assay: BECs, PBMCs, and normal human lung fibroblasts were seeded into 96-well plates (5×10³ cells/well) and treated with Londamocitinib (AZD4604; JAK1-IN-7) (0.001-20 μM) for 72 hours. Cell viability was measured by tetrazolium salt-based assay [2][3] |
| Animal Protocol |
- OVA-induced allergic asthma mouse model: 6-week-old BALB/c mice were sensitized with OVA/alum intraperitoneally on days 0 and 14, then challenged with OVA aerosol (1% w/v) on days 21-27 to induce asthma. Mice were randomly divided into vehicle control and Londamocitinib (AZD4604; JAK1-IN-7) inhalation groups (0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, n=8 per group). The compound was formulated as an aqueous suspension and administered via nebulization once daily for 7 days (days 21-27). On day 28, mice were subjected to AHR measurement, then sacrificed for BAL fluid analysis and lung tissue histopathology [2][3] - Pharmacokinetic animal study: Sprague-Dawley rats and BALB/c mice were administered Londamocitinib (AZD4604; JAK1-IN-7) via inhalation (1 mg/kg) or intravenous injection (0.1 mg/kg). Blood, lung, liver, kidney, and brain samples were collected at 0.25, 0.5, 1, 4, 8, and 24 hours post-dose. Drug concentrations were measured by LC-MS/MS to determine tissue distribution and pharmacokinetic parameters [2] |
| ADME/Pharmacokinetics |
- Absorption: Inhaled Londamocitinib (AZD4604; JAK1-IN-7) is rapidly absorbed in the lungs, with Tmax = 0.5 hours (lung tissue) in mice. Pulmonary bioavailability is 85% following inhalation [2] - Distribution: The compound shows high lung targeting, with lung/plasma concentration ratio of 150 (mice) and 120 (rats) at 1 hour post-inhalation. Minimal distribution to other tissues (liver: lung/liver ratio = 25; kidney: lung/kidney ratio = 30; brain: undetectable) [2] - Metabolism: Londamocitinib (AZD4604; JAK1-IN-7) shows good metabolic stability in human and mouse liver microsomes, with half-lives (t1/2) of 9.5 hours (human) and 8.2 hours (mouse). It is primarily metabolized via oxidative demethylation, with no major toxic metabolites [2][3] - Excretion: In mice, the elimination half-life (t1/2) from lung tissue is 6.8 hours. Approximately 70% of the inhaled dose is excreted in feces and 20% in urine (mainly as parent drug) [2] - Plasma protein binding: The plasma protein binding rate is 95.3 ± 1.2% in human plasma (equilibrium dialysis method) [2][3] |
| Toxicity/Toxicokinetics |
- Acute toxicity: Mice and rats showed no mortality or obvious toxicity symptoms (weight loss, respiratory distress) after a single inhaled dose of Londamocitinib (AZD4604; JAK1-IN-7) up to 10 mg/kg, with maximum tolerated dose (MTD) > 10 mg/kg [2][3] - Subacute toxicity: In mice treated with Londamocitinib (AZD4604; JAK1-IN-7) (1 mg/kg, inhaled, once daily for 28 days), no significant changes were observed in body weight, blood routine parameters (WBC, RBC, PLT), or liver/kidney function indices (ALT, AST, creatinine, urea nitrogen). Histopathological examination of lungs, heart, liver, spleen, and kidneys revealed no abnormal lesions (e.g., inflammation, fibrosis) [2][3] - Pulmonary safety: Inhaled doses up to 1 mg/kg did not induce lung irritation, edema, or epithelial damage in mice. BAL fluid total cell count and inflammatory cytokine levels (TNF-α, IL-6) remained unchanged compared to control [2][3] - Drug interactions: No significant inhibition or induction of CYP450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) was observed at therapeutic concentrations [2] |
| References |
[1]. Jak1 selective inhibitors. WO2018134213A1. [2]. Discovery of the Potent and Selective Inhaled Janus Kinase 1 Inhibitor AZD4604 and Its Preclinical Characterization. J Med Chem. 2023 Oct 12;66(19):13400-13415. [3]. Characterization of Selective and Potent JAK1 Inhibitors Intended for the Inhaled Treatment of Asthma. Drug Des Devel Ther. 2022 Aug 31;16:2901-2917. |
| Additional Infomation |
- Chemical classification: Londamocitinib (AZD4604; JAK1-IN-7) is a small-molecule selective JAK1 inhibitor, belonging to the [pyrazolopyrimidine derivative] class [2][3] - Mechanism of action: The compound binds to the ATP-binding pocket of JAK1, selectively inhibiting its kinase activity. This blocks JAK1-mediated phosphorylation of STAT proteins (STAT3, STAT6) downstream of pro-inflammatory cytokines (IL-4, IL-5, IL-13), suppressing allergic inflammation and airway hyperresponsiveness in asthma [2][3] - Target background: JAK1 is a non-receptor tyrosine kinase that mediates signaling of Th2 cytokines (IL-4, IL-5, IL-13) critical for the pathogenesis of asthma. Aberrant JAK1 activation leads to excessive airway inflammation, eosinophilic infiltration, and mucus hypersecretion [2][3] - Therapeutic potential: Londamocitinib (AZD4604; JAK1-IN-7) is a potent, selective, and inhaled JAK1 inhibitor with lung-targeted delivery and minimal systemic exposure. It shows promising efficacy in preclinical asthma models, with potential applications in the treatment of allergic asthma [2][3] - Administration route: Inhaled formulation (nebulized suspension) designed for local action in the lungs, reducing systemic side effects associated with oral JAK inhibitors [2][3] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~71.43 mg/mL (~119.12 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.17 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 (4.17 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6676 mL | 8.3382 mL | 16.6764 mL | |
| 5 mM | 0.3335 mL | 1.6676 mL | 3.3353 mL | |
| 10 mM | 0.1668 mL | 0.8338 mL | 1.6676 mL |