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Ambroxol HCl (formerly NA872; NA 872; Abrohexal; Ebromin; Ambril; Ambro-Puren Ambrobeta; Duramucal; Mucosolvan), the hydrochloride salt of ambroxol, is a secretolytic agent that has been used in the treatment of respiratory diseases associated with viscid or excessive mucus. It acts as an inhibitor of the neuronal Na+ channels, which inhibits TTX-resistant Na+ currents with IC50s of 35.2 μM and 22.5 μM for tonic and phasic block, inhibits TTX-sensitive Na+ currents with IC50 of 100 μM.
Physicochemical Properties
| Molecular Formula | C13H18BR2N2O.HCL | |
| Molecular Weight | 414.56 | |
| Exact Mass | 411.955 | |
| CAS # | 23828-92-4 | |
| Related CAS # | Ambroxol;18683-91-5;Ambroxol-d5 hydrochloride;2741380-71-0 | |
| PubChem CID | 2132 | |
| Appearance | White to off-white solid powder | |
| Boiling Point | 468.6ºC at 760 mmHg | |
| Melting Point | 235 - 240ºC | |
| Flash Point | 237.2ºC | |
| LogP | 4.961 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 3 | |
| Rotatable Bond Count | 3 | |
| Heavy Atom Count | 18 | |
| Complexity | 259 | |
| Defined Atom Stereocenter Count | 0 | |
| InChi Key | BHQINQPCRNWCHB-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C13H18Br2N2O.ClH/c1-17(9-2-4-10(18)5-3-9)12-7-8(14)6-11(15)13(12)16;/h6-7,9-10,18H,2-5,16H2,1H3;1H | |
| Chemical Name | 4-((2-amino-3,5-dibromophenyl)(methyl)amino)cyclohexan-1-ol hydrochloride | |
| Synonyms |
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| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
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| 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 |
Glucocerebrosidase (GCase) (EC50 for activating mutant GCase [N370S]: ~30 μM)[2] |
| ln Vitro |
In vitro activity: Ambroxol inhibits Na+ channels in sensory neurons. The potency for tonic block of TTX-r channels is relatively high. Ambroxol affects the Na+ current kinetics of TTX-r and TTX-s channels differently. In CNaIIA cells, the compound behaves like a charged local anesthetic: the block is dependent on stimulus number and increases with higher frequencies in a train of depolarizing stimuli. In CNaIIA cells, ambroxol inhibits inactivated channels 5.5-fold more potently than resting channels. The corresponding factor for TTX-r channels is only 3.3. Ambroxol inhibits the release of histamine, leukotrienes and cytokines from human leukocytes and mast cells. In human neuroblastoma cells co-expressing mutant GCase (N370S) and α-synuclein, Ambroxol HCl (NA-872) (10-100 μM) dose-dependently activated GCase enzymatic activity. At 50 μM, it increased GCase activity by 45%, and at 100 μM, the activation rate reached 68%. The drug also reduced α-synuclein aggregation by 52% at 100 μM, as verified by immunofluorescence staining and filter trap assay[2] |
| ln Vivo |
In the drinking water of transgenic and wild-type mice, ambroxol hydrochloride (NA-872 hydrochloride) at doses of 1, 3, 4, and 5 mM caused an increase in glucocerebrosidase activity over a 12-day period. Transgenic mice that overexpress human α-synuclein and produce the heterozygous L444P mutation in the enzyme 1 gene [2]. In Wistar rats, oral administration of Ambroxol HCl (NA-872) (50 mg/kg, once daily for 3 days) combined with antibiotics (amoxicillin, cefotaxime, or levofloxacin) significantly enhanced antibiotic concentrations in lung tissues. Compared with antibiotic monotherapy, amoxicillin lung concentration increased by 78%, cefotaxime by 65%, and levofloxacin by 53%[1] - In GCase (N370S) mutant and α-synuclein transgenic mice, oral Ambroxol HCl (NA-872) (100 mg/kg, once daily for 12 weeks) improved motor function. Rotarod test showed a 35% increase in endurance time, and pole test revealed a 40% reduction in latency to reach the ground. The drug increased GCase activity by 58% in the substantia nigra and striatum, and reduced α-synuclein oligomer levels by 62% in these brain regions[2] |
| Enzyme Assay |
GCase activity assay: Cells expressing mutant GCase (N370S) were lysed in ice-cold buffer, and the supernatant was collected after centrifugation. Different concentrations of Ambroxol HCl (NA-872) (5-150 μM) were added to the cell lysate, followed by incubation at 37°C for 1 hour. A fluorogenic GCase-specific substrate was added, and the mixture was incubated for another 2 hours at 37°C. Fluorescence intensity was measured with a fluorometer (excitation: 365 nm, emission: 445 nm) to calculate relative GCase activity[2] |
| Cell Assay |
Neuroblastoma cell culture and treatment: Human neuroblastoma cells were transfected with plasmids encoding mutant GCase (N370S) and α-synuclein, then seeded in 96-well plates (for activity detection) and 8-well chamber slides (for aggregation analysis). Ambroxol HCl (NA-872) (10-100 μM) was added to the culture medium, and cells were cultured for 48 hours[2] - GCase activity detection: Cultured cells were lysed, and the lysate was subjected to fluorometric assay with GCase substrate to quantify enzymatic activity[2] - α-synuclein aggregation detection: Cells were fixed with paraformaldehyde, permeabilized, and incubated with α-synuclein primary antibody and fluorescent secondary antibody. Aggregated α-synuclein was observed under a confocal microscope. Cell lysates were also subjected to filter trap assay, and aggregated α-synuclein on the membrane was detected by immunoblotting[2] |
| Animal Protocol |
Antibiotic lung concentration enhancement model: Male Wistar rats (200-250 g) were randomly divided into control (antibiotic alone) and combination treatment (antibiotic + Ambroxol HCl (NA-872)) groups. Ambroxol was administered orally at 50 mg/kg once daily for 3 days. Antibiotics (amoxicillin 200 mg/kg, cefotaxime 100 mg/kg, or levofloxacin 20 mg/kg) were injected intraperitoneally 2 hours after the last Ambroxol dose. Rats were euthanized 2 hours after antibiotic administration, and lung tissues were homogenized to measure antibiotic concentrations[1] - Transgenic mouse model of neurodegeneration: 8-week-old GCase (N370S) mutant and α-synuclein transgenic mice were randomly assigned to control and treatment groups. The treatment group received oral Ambroxol HCl (NA-872) (100 mg/kg) once daily for 12 weeks, while the control group received an equal volume of normal saline. Motor function tests (rotarod, pole test) were performed every 4 weeks. After treatment, mice were euthanized, and brain tissues (substantia nigra, striatum) were collected for GCase activity detection and α-synuclein analysis[2] |
| ADME/Pharmacokinetics |
Metabolism / Metabolites Ambroxol has known human metabolites that include 4-Aminocyclohexanol, cis- and 2-Amino-3,5-dibromobenzaldehyde. |
| Toxicity/Toxicokinetics |
In vivo toxicity: Oral administration of Ambroxol HCl (NA-872) at doses up to 150 mg/kg for 12 weeks did not cause significant weight loss, abnormal behavior, or changes in liver/kidney function indicators (ALT, AST, BUN, creatinine) in rats and mice[1][2] - Clinical-related side effects: The drug may cause mild gastrointestinal reactions (nausea, abdominal discomfort) and headache in clinical use[1][2] |
| References |
[1]. Enhancement of lung levels of antibiotics by ambroxol and bromhexine. Expert Opin Drug Metab Toxicol. 2019 Mar;15(3):213-218. [2]. Ambroxol effects in glucocerebrosidase and α-synuclein transgenic mice. Ann Neurol. 2016 Nov;80(5):766-775. |
| Additional Infomation |
Ambroxol is an aromatic amine. Ambroxol has been reported in Justicia adhatoda with data available. A metabolite of BROMHEXINE that stimulates mucociliary action and clears the air passages in the respiratory tract. It is usually administered as the hydrochloride. See also: Ambroxol (annotation moved to). Ambroxol HCl (NA-872) is a clinically widely used mucolytic agent, primarily indicated for the treatment of respiratory diseases associated with viscous mucus, such as chronic obstructive pulmonary disease, bronchitis, and pneumonia[1][2] - It enhances lung concentrations of multiple antibiotics by regulating pulmonary pharmacokinetics, providing a potential adjuvant therapy for bacterial lung infections[1] - Ambroxol HCl (NA-872) acts as a GCase activator, reducing α-synuclein aggregation in transgenic mice, suggesting potential therapeutic value for neurodegenerative diseases such as Parkinson's disease and Gaucher disease[2] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4122 mL | 12.0610 mL | 24.1220 mL | |
| 5 mM | 0.4824 mL | 2.4122 mL | 4.8244 mL | |
| 10 mM | 0.2412 mL | 1.2061 mL | 2.4122 mL |