Quinolone

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].

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].

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.

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.

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.

[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.

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.)