Quinolone
Bacterial DNA gyrase (topoisomerase II) and topoisomerase IV; pannexin 1 (PANX1) channel (IC50 = 4 microM for PANX1 inward current).

HepG2 cells exposed to trovafloxacin (20 µM; 24 hours) and tumor necrosis factor (TNF; 4 ng/mL) exhibit increased lactate dehydrogenase (LDH) leakage and apoptosis.After incubating HepG2 cells with trovafloxacin (20 µM) for 24 hours and TNF (4 ng/mL), the expression of early NF-κB-related factors A20 and IκBα is increased.In HepG2, trovafloxacin prolongs TNF-induced MAPK activation and IKKα/β activation[1].
Effectively preventing apoptotic cells from absorbing TO-PRO-3 is trovafloxacin. Moreover, trovafloxacin prevents apoptotic cells from releasing ATP. Trovafloxacin does not prevent PANX1 cleavage during apoptosis or caspase 3/7 activation[2].
With MICs of 0.06-0.25 mg/mL recorded for over 700 isolates, trovafloxacin is equally effective against pneumococci that are susceptible to penicillin as well as those that are resistant to it. Trovafloxacin's minimum inhibitory concentration (MIC) for 90% of pneumococci isolates is 0.125 μg/mL [3].

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Trovafloxacin is a broad-spectrum quinolone antibiotic that blocks DNA supercoiling in various bacteria by inhibiting the activity of DNA gyrase (topoisomerase II) and topoisomerase IV. It has potent activity against Gram-positive, Gram-negative, and anaerobic organisms. The MIC90 for 90% of Streptococcus pneumoniae isolates is 0.125 microg/mL. Trovafloxacin is equally effective against penicillin-sensitive and penicillin-resistant pneumococci. It inhibits PANX1 (pannexin 1) channels with an IC50 of 4 microM for PANX1 inward current, but does not inhibit connexin 43 gap junctions or PANX2. In HepG2 human hepatocytes, exposure to trovafloxacin (20 microM; 24 hours) and TNF (4 ng/mL) increases lactate dehydrogenase (LDH) leakage and apoptosis. Incubation of HepG2 cells with trovafloxacin (20 microM) and TNF (4 ng/mL) for 24 hours increases the expression of early NF-kappaB-related factors A20 and IkappaBalpha. In HepG2 cells, trovafloxacin prolongs TNF-induced MAPK activation and IKKalpha/beta activation. Trovafloxacin also effectively prevents apoptotic uptake of TO-PRO-3 and prevents the release of ATP from apoptotic cells, consistent with PANX1 inhibition. Trovafloxacin does not prevent apoptosis or PANX1 cleavage during caspase 3/7 activation. Trovafloxacin exhibits hepatocyte toxicity in vitro, which correlates with its clinical hepatotoxicity.

Treatment with trovafloxacin (150 mg/kg; oral; male C57BL/6 J mice) prevents the nuclear translocation of p65 that is induced by TNF. Treatment with trovafloxacin increases the expression of IκBα and A20, early NF-κB-related factors[1].When trovafloxacin is given to mice along with lipopolysaccharide (LPS) or tumor necrosis factor (TNF), it causes severe liver toxicity that is accompanied by large areas of the liver that are apoptotic, elevated serum levels of alanine amino transferases (ALT), and pro-inflammatory cytokines[1].

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Trovafloxacin demonstrates in vivo antibacterial efficacy in mouse models of bacterial infection. In addition, trovafloxacin (150 mg/kg; oral; male C57BL/6J mice) prevents TNF-induced p65 nuclear translocation, increases the expression of early NF-kappaB-related factors IkappaBalpha and A20, and prevents TNF-induced p65 nuclear translocation. When administered to mice with LPS or TNF, trovafloxacin causes severe hepatotoxicity, accompanied by extensive liver apoptosis, elevated serum alanine aminotransferase (ALT), and hepatic necrosis. This hepatotoxicity is mouse strain-specific and TNF-dependent, and has been used as a model of idiosyncratic drug-induced liver injury (DILI). In a mouse model of pancreatitis, trovafloxacin (through PANX1 inhibition) reduces disease severity. Trovafloxacin has also been used in research as a tool to study PANX1 function in inflammation, cell death, and ATP release. The antibacterial effects are standard for fluoroquinolone antibiotics.

The antibacterial activity of trovafloxacin is measured using a standard broth microdilution assay to determine the minimum inhibitory concentration (MIC). Bacteria (e.g., S. aureus, E. coli, S. pneumoniae) are grown overnight in Mueller-Hinton broth (supplemented with 5% lysed horse blood for S. pneumoniae). A bacterial suspension is prepared and adjusted to a concentration of 5×10⁵ CFU/mL. Trovafloxacin is dissolved in DMSO and serially diluted (2-fold) in the growth medium in 96-well plates, typically over a concentration range of 0.001-128 microg/mL. The bacterial suspension is added to each well, and the plates are incubated at 35-37degC for 18-24 hours. The MIC is defined as the lowest concentration of trovafloxacin that completely inhibits visible bacterial growth. For PANX1 channel inhibition, a whole-cell patch clamp electrophysiology assay is used. HEK-293 cells stably expressing human PANX1 are seeded on coverslips. Whole-cell currents are recorded using an Axopatch 200B amplifier. The extracellular solution contains (in mM): 140 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 10 HEPES, 10 glucose (pH 7.4). The pipette solution contains (in mM): 130 CsCl, 10 NaCl, 2 MgCl2, 1 CaCl2, 10 HEPES, 10 EGTA, 2 ATP (pH 7.2). Trovafloxacin is applied by perfusion at varying concentrations (0.1-100 microM). PANX1 channels are activated by depolarizing voltage steps to +50 mV. The inward current amplitude is measured, and the percentage inhibition is calculated. The IC50 is calculated by fitting the concentration-response curve. For hepatocyte toxicity, primary mouse or human hepatocytes are treated with trovafloxacin (0-1000 microM) with or without TNF (4 ng/mL) for 24-48 hours, and cell viability is measured by LDH release assay.

The antibacterial activity of trovafloxacin is measured using a standard broth microdilution assay to determine the minimum inhibitory concentration (MIC). Bacteria (e.g., S. aureus, E. coli, S. pneumoniae) are grown overnight in Mueller-Hinton broth (supplemented with 5% lysed horse blood for S. pneumoniae). A bacterial suspension is prepared and adjusted to a concentration of 5×10⁵ CFU/mL. Trovafloxacin is dissolved in DMSO and serially diluted (2-fold) in the growth medium in 96-well plates, typically over a concentration range of 0.001-128 microg/mL. The bacterial suspension is added to each well, and the plates are incubated at 35-37degC for 18-24 hours. The MIC is defined as the lowest concentration of trovafloxacin that completely inhibits visible bacterial growth. For cytotoxicity in eukaryotic cells, HepG2 human hepatocytes or primary mouse hepatocytes are used. Cells are seeded in 96-well plates at 1-2×10⁴ cells per well in DMEM with 10% FBS. After overnight attachment, the medium is replaced with fresh medium containing varying concentrations of trovafloxacin (0.1-1000 microM, 3-fold serial dilutions, prepared in DMSO, final DMSO ≤0.5%). Some wells are also treated with TNF (4 ng/mL) to potentiate hepatotoxicity. Control wells receive DMSO alone or vehicle. Plates are incubated for 24-48 hours at 37degC. Cell viability is measured by LDH release assay (measure LDH in supernatant using a commercial kit) or by MTT assay. The CC50 (50% cytotoxic concentration) is calculated. Apoptosis is assessed by measuring caspase-3/7 activity using a luminescent assay, or by annexin V/PI staining and flow cytometry. NF-kappaB pathway activation is assessed by Western blot for IkappaBalpha degradation and p65 nuclear translocation, or by luciferase reporter assay. For PANX1 inhibition in cells, ATP release from apoptotic cells is measured: cells are induced to undergo apoptosis with staurosporine (1 microM) or TNF plus cycloheximide, and then trovafloxacin (10-100 microM) is added; ATP in the supernatant is measured using a bioluminescent ATP assay kit.

Animal Model: Male C57BL/6 J mice (9-11-week-old) injected with recombinant murine TNF ion[1]
Dosage: 150 mg/kg
Administration: Oral administration
Result: revealed a higher proportion of cells in the liver with an elevated nuclear/cytoplasmic p65 ratio.

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The in vivo antibacterial efficacy of trovafloxacin is evaluated in mouse models of systemic infection. Female CD-1 mice (18-20 g) are infected intraperitoneally with a lethal dose of bacteria (e.g., S. aureus, S. pneumoniae, E. coli) in 0.5 mL of sterile saline containing 5% hog gastric mucin (to enhance virulence). One hour post-infection, mice are treated with trovafloxacin (oral, subcutaneous, or intravenous) at various doses (e.g., 1, 3, 10, 30, 100 mg/kg). Survival is monitored for 7 days, and the ED50 (50% effective dose) is calculated. The hepatotoxicity model is a key in vivo protocol for studying trovafloxacin-induced idiosyncratic liver injury. Male C57BL/6J mice (6-8 weeks old, 20-25 g) are given a single oral dose of trovafloxacin (150 mg/kg) suspended in 0.5% methylcellulose. Some groups may also receive LPS (0.2-2 mg/kg, i.p.) or TNF (1-10 microg/mouse, i.v.) to potentiate hepatotoxicity. Control mice receive vehicle alone or LPS/TNF alone. Blood is collected at 6, 12, 24, 48 hours post-dosing for serum ALT, AST, and bilirubin measurement. Mice are euthanized at 24 hours, and livers are excised, weighed, and fixed in 10% formalin for histopathology (H&E staining) to assess necrosis, inflammation, and apoptosis (TUNEL staining). Serum levels of pro-inflammatory cytokines (TNF, IL-6, IL-1beta) are measured by ELISA. Trovafloxacin-treated mice (especially those co-treated with LPS or TNF) develop severe hepatitis with high ALT levels and extensive hepatocellular necrosis. This model is widely used to study the mechanisms of idiosyncratic drug-induced liver injury.

Absorption, Distribution and Excretion
After oral administration, this product is well absorbed in the gastrointestinal tract and is unaffected by food intake. The absolute bioavailability is approximately 88%. Approximately 50% of the oral dose is excreted unchanged (43% in feces and 6% in urine). Metabolism/Metabolites Metabolism: Travafloxacin is primarily metabolized via conjugation (cytochrome P450 oxidative metabolism has minimal effect on travafloxacin). Major metabolites include ester glucuronides, mainly found in urine (13% of the administered dose); and N-acetyl metabolites, mainly found in feces and serum (9% and 2.5% of the administered dose, respectively). Other minor metabolites include diacids, hydroxycarboxylic acids, and aminosulfonic acids, which are detected in trace amounts in feces and urine (<4% of the administered dose).
The known human metabolites of trovafloxacin include (2S,3S,4S,5R)-6-[7-[(1R,5S)-6-amino-3-azabicyclo[3.1.0]hexane-3-yl]-1-(2,4-difluorophenyl)-6-fluoro-4-oxo-1,8-naphthidine-3-carbonyl]oxy-3,4,5-trihydroxyoxacyclohexane-2-carboxylic acid.

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Biological half-life
After oral administration, the half-life is 9.1 hours to 12.2 hours in the dose range of 100 to 200 mg tablets. After intravenous infusion, the half-life is 9.4 to 12.7 hours in the dose range of 100 to 300 mg.

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The pharmacokinetics (PK) of trovafloxacin have been well characterized in animals and humans. In humans, following oral administration, trovafloxacin is rapidly absorbed with an absolute bioavailability of approximately 90%. The time to peak concentration (Tmax) is 1-2 hours. The plasma elimination half-life (t1/2) is long, approximately 10-12 hours, allowing once-daily dosing. The volume of distribution (Vd) is large ( > 2 L/kg), indicating extensive tissue distribution, including penetration into the lung, bone, and prostate. Protein binding is moderate (approximately 70%). The clearance (CL) is primarily via hepatic metabolism (glucuronidation) and biliary excretion; renal excretion is a minor route (< 10% excreted unchanged in urine). In rats, the half-life is shorter (2-4 hours), and the oral bioavailability is moderate to high. The PK of trovafloxacin is dose-proportional over a range of 100-800 mg in humans. The compound can cross the blood-brain barrier. Due to its hepatotoxicity, trovafloxacin is no longer widely used; however, it is used as a research tool.

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Currently, there is no clinical information regarding the use of trovafloxacin during lactation; however, the drug concentration in breast milk appears to be very low. Traditionally, fluoroquinolones are not recommended for use in infants due to concerns about adverse effects on the developing joints of infants. However, recent studies suggest the risk is minimal. Calcium in breast milk may prevent the absorption of small amounts of fluoroquinolones in breast milk, but there is currently insufficient data to confirm or refute this claim. Lactating women can use trovafloxacin, but monitoring for potential impacts on the infant's gut microbiota, such as diarrhea or candidiasis (thrush, diaper rash), is necessary. However, alternative medications with available safety information are preferred.
◉ Effects on Breastfed Infants
No published information found as of the revision date.
◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.

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Protein binding
The average plasma protein binding rate is approximately 76%, and it is independent of concentration.

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Trovafloxacin is known for its severe hepatotoxicity, which led to its withdrawal from many markets and severe restriction of its use. In clinical trials and post-marketing surveillance, trovafloxacin caused acute liver injury, including fulminant liver failure requiring transplantation and death. The incidence of severe liver injury was approximately 1 in 10,000-30,000 patients, which is higher than other fluoroquinolones. The hepatotoxicity is idiosyncratic (non-dose-dependent) and is associated with the compound's ability to inhibit mitochondrial function, deplete glutathione, and activate the TNF pathway. In the mouse model, trovafloxacin (150 mg/kg) with LPS or TNF co-administration causes severe hepatotoxicity (ALT > 5000 U/L, massive necrosis), while the compound alone at clinically relevant doses does not cause significant injury in mice, reflecting the idiosyncratic nature. Common adverse effects of trovafloxacin (aside from hepatotoxicity) include gastrointestinal disturbances (nausea, vomiting, diarrhea), central nervous system effects (headache, dizziness, seizures), and photosensitivity. Fluoroquinolones as a class are associated with an increased risk of tendinitis and tendon rupture, QT prolongation (arrhythmia risk), and peripheral neuropathy. For laboratory handling, trovafloxacin should be handled with standard chemical safety precautions: use gloves, lab coat, eye protection.

[1]. The hepatotoxic fluoroquinolone trovafloxacin disturbs TNF- and LPS-induced p65 nuclear translocation in vivo and in vitro. Toxicol Appl Pharmacol. 2020 Mar 15;391:114915.

[2]. Unexpected link between an antibiotic, pannexin channels and apoptosis. Nature. 2014 Mar 20;507(7492):329-34.

[3]. Activity of the new fluoroquinolone trovafloxacin (CP-99,219) against DNA gyrase and topoisomerase IV mutants of Streptococcus pneumoniae selected in vitro. Antimicrob Agents Chemother. 1996 Dec;40(12):2691-7.

Travafloxacin is a 1,8-naphthidine derivative with the structure 4-oxo-1,4-dihydro-1,8-naphthidine-3-carboxylic acid, with 2,4-difluorophenyl, fluorine, and 6-amino-3-azabicyclo[3.1.0]hex-3-yl substituents at positions 1, 6, and 7, respectively. It was a broad-spectrum antibiotic that was withdrawn from the market due to the risk of liver failure. Travafloxacin possesses multiple functions, including antibacterial, hepatotoxic, topoisomerase IV inhibitor, DNA synthesis inhibitor, and antiviral activity. It is a 1,8-naphthidine derivative, amino acid, monocarboxylic acid, azabicycloalkane, tertiary amine compound, primary amine compound, quinolone antibiotic, fluoroquinolone antibiotic, and difluorobenzene compound. It is the conjugate base of travafloxacin (1+). Travafloxacin is a broad-spectrum antibiotic, formerly marketed by Pfizer under the brand name Trovan. It exerts its antibacterial effect by inhibiting the unwinding of supercoiled DNA in various bacteria by inhibiting the activity of DNA gyrase and topoisomerase IV. Compared to previous fluoroquinolones, it is more effective against Gram-positive bacteria than against Gram-negative bacteria. Due to its hepatotoxicity, trovafloxacin has been withdrawn from the market. Drug Indications For the treatment of infections caused by susceptible strains of specified microorganisms, including uncomplicated urethral gonorrhea in men and cervical and rectal gonorrhea in women caused by Neisseria gonorrhoeae, as well as non-gonococcal urethritis and cervicitis caused by Chlamydia trachomatis. Trovafloxacin is a synthetic broad-spectrum quinolone antibacterial agent indicated for the treatment of the following infections in adults: Pneumonia: community-acquired pneumonia and hospital-acquired pneumonia (mild, moderate, and severe). Note: Its efficacy in patients with severe hospital-acquired pneumonia, particularly infections caused by less susceptible pathogens such as Pseudomonas aeruginosa, has not been established. See also Section 4.2. Acute exacerbations of chronic bronchitis, acute sinusitis, complicated intra-abdominal infections and acute pelvic infections, salpingitis, uncomplicated gonococcal urethritis and cervicitis, chlamydial cervicitis, and complicated skin and soft tissue infections. Official guidelines for the rational use of antimicrobial agents should be considered. Travafloxacin is a synthetic broad-spectrum quinolone antibiotic indicated for the treatment of the following infections in adults: Pneumonia: community-acquired pneumonia and hospital-acquired pneumonia (mild, moderate, and severe). Note: Its efficacy in patients with severe hospital-acquired pneumonia, particularly infections caused by less susceptible pathogens such as Pseudomonas aeruginosa, has not been established. See also Section 4.2. Acute exacerbations of chronic bronchitis, acute sinusitis, complicated intra-abdominal infections and acute pelvic infections, salpingitis, uncomplicated gonococcal urethritis and cervicitis, chlamydial cervicitis, and complicated skin and soft tissue infections. Official guidelines for the rational use of antimicrobial agents should be considered. Mechanism of Action Travafloxacin is a fluoronaphthidine ketone, associated with fluoroquinolones, and is active in vitro against a variety of Gram-negative and Gram-positive aerobic and anaerobic bacteria. Its bactericidal action stems from its inhibition of DNA gyrase and topoisomerase IV. DNA gyrase is an important enzyme involved in bacterial DNA replication, transcription, and repair. Topoisomerase IV is an enzyme that plays a crucial role in chromosomal DNA allocation during bacterial cell division. Pharmacodynamics Travafloxacin is a broad-spectrum antibiotic that inhibits DNA supercoiling in various bacteria by blocking the activity of DNA gyrase and topoisomerase IV. Due to the risk of hepatotoxicity, it is not widely used. Compared to previous fluoroquinolones, it exhibits stronger antibacterial activity against Gram-positive bacteria but weaker antibacterial activity against Gram-negative bacteria. The mechanisms of action of fluoroquinolones, including trovafloxacin, differ from those of penicillins, cephalosporins, aminoglycosides, macrolides, and tetracyclines. Therefore, fluoroquinolones may be effective against pathogens resistant to these antibiotics. No cross-resistance exists between trovafloxacin and the aforementioned antibiotic classes. Overall results from in vitro synergistic studies (testing combinations of trovafloxacin with β-lactam and aminoglycoside antibiotics) indicate that synergistic effects are strain-specific and uncommon. This is consistent with previous results obtained using other fluoroquinolones. In vitro resistance to trovafloxacin develops slowly through multiple mutations, similar to other fluoroquinolones. The frequency of in vitro resistance to trovafloxacin is typically between 1 × 10⁻⁷ and 10⁻¹⁰. Although cross-resistance has been observed between trovafloxacin and some other fluoroquinolones, some microorganisms resistant to other fluoroquinolones may be susceptible to trovafloxacin.

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The compound is for research use only. Store at -20degC, protected from light. Trovafloxacin (CP-99219) is a broad-spectrum quinolone antibiotic that inhibits DNA gyrase and topoisomerase IV, leading to bacterial DNA damage and cell death. It was approved for the treatment of community-acquired pneumonia, nosocomial pneumonia, chronic bronchitis, and uncomplicated skin infections. However, due to severe hepatotoxicity (liver injury), its use is now restricted to only the most serious infections, and it has been withdrawn from many countries (including the US, Europe). Trovafloxacin is also a potent inhibitor of the pannexin 1 (PANX1) channel (IC50 = 4 microM), which mediates ATP release during apoptosis and inflammation. Trovafloxacin is used as a research tool to study PANX1 function in apoptosis, inflammation, and cell death. The compound has been used to study the role of PANX1 in cancer, atherosclerosis, and neuroinflammation. Trovafloxacin is commercially available for research use only. It should be stored at -20degC and protected from light. Due to its hepatotoxicity, extreme caution should be taken when handling this compound. It is not for human use.

C20H15F3N4O3

416.35

416.11

147059-72-1

Trovafloxacin mesylate;147059-75-4

62959

White to light yellow solid powder

1.612g/cm3

630.5ºC at 760mmHg

246ºC

335.1ºC

9.21E-17mmHg at 25°C

1.672

2.659

2

10

3

30

770

2

C1[C@@H]2[C@@H](C2N)CN1C3=C(C=C4C(=O)C(=CN(C4=N3)C5=C(C=C(C=C5)F)F)C(=O)O)F

WVPSKSLAZQPAKQ-SOSAQKQKSA-N

InChI=1S/C20H15F3N4O3/c21-8-1-2-15(13(22)3-8)27-7-12(20(29)30)17(28)9-4-14(23)19(25-18(9)27)26-5-10-11(6-26)16(10)24/h1-4,7,10-11,16H,5-6,24H2,(H,29,30)/t10-,11+,16

7-[(1R,5S)-6-amino-3-azabicyclo[3.1.0]hexan-3-yl]-1-(2,4-difluorophenyl)-6-fluoro-4-oxo-1,8-naphthyridine-3-carboxylic acid

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)

DMSO : ~9.09 mg/mL (~21.83 mM)

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.)