Absorption, Distribution and Excretion
Orally administered benzalkonium chloride is well absorbed, with plasma drug concentrations rapidly reaching peak levels before declining, and a half-life of approximately 13 hours. With topical application, although local drug concentrations are relatively high, systemic absorption of benzalkonium chloride is relatively low compared to oral doses. This lower local absorption helps reduce the likelihood of systemic drug side effects with topical administration of benzalkonium chloride. The relatively high lipid solubility of the weakly basic benzalkonium chloride is thought to be related to significant passive reabsorption in the renal tubules, suggesting that only about 5% of benzalkonium chloride is excreted unchanged in the urine. However, other studies have shown that a considerable proportion (50-65%) of the drug is excreted unchanged in the urine. Several inactive oxidative metabolites of benzalkonium chloride are excreted in the urine, while benzalkonium chloride N-oxide metabolites remain in the plasma for a longer period, with a longer half-life than the parent compound benzalkonium chloride. However, it is generally accepted that benzalkonium chloride is primarily excreted in the urine, mostly as inactive metabolites or conjugates. The volume of distribution of benzalkonium chloride is 10 liters. The systemic clearance of benzalkonium is 170 mL/min. Benzamin is primarily metabolized via oxidation, dealkylation, and conjugation to hydroxyl metabolites, dealkylated metabolites, and N-oxide metabolites. However, generally, when using the recommended dose, the levels of benzalkonium absorbed or exposed in the body are usually insufficient to produce systemic pharmacological effects [L

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biological half-life
approximately 13 hours after oral administration, with a terminal half-life of approximately 7.7 hours].

Protein Binding

After oral administration of benzalkonium chloride, the plasma protein binding rate is <20%.

[1]. Quane PA, Pharmacology of benzydamine. Inflammopharmacology. 1998;6(2):95-107

Benzafim belongs to the indazole class of compounds, with a benzyl group at position 1 and a 3-(dimethylamino)propyl group at positions 3. It is a locally acting nonsteroidal anti-inflammatory drug (NSAID) with both local anesthetic and analgesic effects. Benzafim can be used as a central nervous system stimulant, NSAID, hallucinogen, local anesthetic, and analgesic. It belongs to the indazole class, aromatic ethers, and tertiary amines. It is the conjugate base of benzazine (1+). Benzafim (also known as Tantum Verde or Difflam), existing as a hydrochloride salt, is a locally acting NSAID with local anesthetic and analgesic effects. It is used for the local relief of pain and inflammation in the oral cavity, throat, or musculoskeletal system. Although benzalkonium chloride, an indazole analogue, is a nonsteroidal anti-inflammatory drug (NSAID), it possesses several physicochemical properties and pharmacological activities distinct from traditional aspirin-like NSAIDs. These characteristics contribute to benzalkonium chloride's mechanism of action as an effective locally acting NSAID, providing local anesthetic and analgesic effects. Furthermore, unlike acidic or acid-metabolized aspirin-like NSAIDs, benzalkonium chloride is actually a weak base. Benzaminium chloride is a benzylindazole with analgesic, antipyretic, and anti-inflammatory effects. It is used to reduce postoperative and post-traumatic pain and edema and to promote wound healing. It can also be used topically to treat rheumatic diseases and inflammation of the oral cavity and throat.

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Drug Indications
Benzaminium is primarily available as mouthwash, oral mucosal spray, or topical cream, and is most commonly used for local analgesia and anti-inflammation to relieve painful inflammation.
When formulated as a mouthwash or spray, benzalkonium chloride can be used to treat traumatic conditions, such as pharyngitis following tonsillectomy or nasogastric tube use; inflammatory conditions, such as pharyngitis, recurrent aphthous ulcers, and oral ulcers resulting from radiation therapy, dental surgery, and procedures; or more common conditions, such as sore throat, glossitis, gingivitis, oral ulcers, or discomfort caused by dentures. When used as a topical cream, benzalkonium chloride can be used to relieve symptoms associated with painful inflammation of the musculoskeletal system, including acute inflammatory conditions (such as myalgia and bursitis) or traumatic conditions (such as sprains, strains, bruises, muscle aches, joint stiffness, and even post-fracture sequelae).

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Mechanism of Action
Although benzalkonium chloride is classified as a nonsteroidal anti-inflammatory drug (NSAID), its mechanism of action differs from that of traditional aspirin-like NSAIDs.
Specifically, benzalkonium chloride primarily exerts its effects by inhibiting the synthesis of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), while having less effect on other pro-inflammatory cytokines such as IL-6 and IL-8, or anti-inflammatory cytokines such as IL-10 or IL-1 receptor antagonists. Furthermore, benzalkonium chloride exhibits weak inhibition of prostaglandin synthesis, as studies have shown that it only effectively inhibits the activity of cyclooxygenase (COX) and lipoxygenase at concentrations of 1 mM or higher. Given that benzalkonium chloride is currently mostly used topically and is generally poorly absorbed by the skin and/or nonspecific mucous membranes, it often fails to reach the absorption or blood concentrations required to produce any distal systemic effects or inhibit COX, thus limiting its action to a localized level. In addition, some studies hypothesize that benzalkonium chloride can inhibit oxidative burst and membrane stability in neutrophils. These effects are manifested in its ability to inhibit neutrophil granule release and stabilize lysosomes. In addition, benzalkonium chloride also has a local anesthetic effect, which may be related to its ability to inhibit the release of inflammatory mediators (such as substance P and calcitonin gene-related peptide) from sensory nerve endings. Since substance P can induce histamine release from mast cells, benzalkonium chloride's inhibition of substance P release further enhances its anti-inflammatory effect. At a concentration of approximately 3 mmol/L, benzalkonium chloride also exhibits non-specific antibacterial activity against various broad-spectrum antibiotic-resistant bacterial strains (such as ampicillin, chloramphenicol, and tetracycline). When used in combination with other antibiotics (such as tetracycline and chloramphenicol), benzalkonium chloride also has a synergistic effect against antibiotic-resistant Staphylococcus aureus and Pseudomonas aeruginosa strains.

C19H23N3O

309.41

309.184

642-72-8

132-69-4 (mono-hydrochloride)

12555

Typically exists as solid at room temperature

1.1±0.1 g/cm3

474.4±35.0 °C at 760 mmHg

320°F

240.7±25.9 °C

0.0±1.2 mmHg at 25°C

1.580

3.78

0

3

7

23

344

0

CN(C)CCCOC1=NN(CC2=CC=CC=C2)C3=CC=CC=C31

CNBGNNVCVSKAQZ-UHFFFAOYSA-N

InChI=1S/C19H23N3O/c1-21(2)13-8-14-23-19-17-11-6-7-12-18(17)22(20-19)15-16-9-4-3-5-10-16/h3-7,9-12H,8,13-15H2,1-2H3

3-(1-benzylindazol-3-yl)oxy-N,N-dimethylpropan-1-amine

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.

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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).

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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)

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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).

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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

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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.

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