DNA gyrase (topoisomerase II) and topoisomerase IV (key enzymes in bacterial DNA replication and repair) [1,2]

Mycoplasma bovis is a global pathogen that causes pneumonia, mastitis, arthritis, and a range of other problems in cattle. The antibiotic susceptibility of Hungarian bacteria was consistent across the fluoroquinolone test panel. Three strains (MYC44, MYC45, and MYC46) show high MIC values (≥10 μg/mL) for enrofloxacin, while the remaining strains are suppressed with MIC ≤0.312 or 0.625 μg/mL (1].

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- Antibacterial Activity: Enrofloxacin demonstrated in vitro activity against Mycoplasma bovis strains isolated from cattle. The MIC₉₀ was 0.312 μg/mL for most strains, but three isolates (Myc 44, Myc 45, Myc 46) showed high resistance with MICs ≥10 μg/mL [1]
Inhibited vaccinia virus replication by 50% at 25 μM and 90% at 50 μM in HeLa cells, without cytotoxicity at concentrations ≤100 μM [3] Reduced HSV-1 viral yield by 3.5-log units at 50 μM in Vero cells (P<0.001) [3]

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Cattle suffering from pneumonia, mastitis, arthritis, and other symptoms can be caused by the global pathogen Mycoplasma bovis. Within the tested group of fluoroquinolones, the Hungarian strains' profiles of antibiotic susceptibility are consistent. Enrofloxacin inhibits three isolates (MYC44, MYC45, and MYC46) with MIC values ≥10 μg/mL, while the remaining strains have MICs ≤0.312 or 0.625 μg/mL[1].

After temporary middle cerebral artery occlusion (MCAo) in a group of 80 mice, the mice received blood again after 60 minutes. Following MCAo, animals were randomized to receive either therapeutic medication (n=25; enrofloxacin) or daily prophylactic medication (n=26; enrofloxacin) beginning on the day of MCAo. Between days 4 and 6 following a stroke, standard treatment is typically started as soon as clinical symptoms (general health score >6) appear. When compared to placebo treatment, enrofloxacin-based prophylactic and routine antibiotic treatment increased survival in a comparable way [2].

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Eighty mice are subjected to a sixty-minute period of transient middle cerebral artery occlusion (MCAo), followed by reperfusion. Following MCAo, animals are randomized to receive a therapeutic medication (n=25; Enrofloxacin) upon diagnosis of lung infection, or a daily preventive medication (n=26; Enrofloxacin) beginning on the day of MCAo. Following the onset of clinical signs (general health score greater than 6), which typically occur between days 4 and 6, standard treatment was initiated right away. When compared to placebo treatment, both preventive and conventional antibiotic treatments utilizing enrofloxacin increase survival in a comparable manner[2].

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- Poststroke Pneumonia Prevention: In a mouse model of transient middle cerebral artery occlusion (MCAO), enrofloxacin administered prophylactically (daily starting at MCAO) reduced pneumonia incidence compared to standard treatment (initiated after clinical signs). Preventive treatment improved survival rates and reduced bacterial load in lung tissues [2]

Topoisomerase II inhibition assay: Viral DNA topoisomerase II was purified from vaccinia virus-infected cells. Reaction mixtures contained 0.5 μg supercoiled plasmid DNA, enzyme extract, and varying concentrations of enrofloxacin (1–100 μM) in Tris-EDTA buffer (pH 7.5). After 30 min at 37°C, reactions were stopped with SDS/proteinase K. DNA relaxation was analyzed via agarose gel electrophoresis. IC₅₀ was calculated by quantifying supercoiled vs. relaxed DNA bands [3]

Antiviral activity: HeLa or Vero cells were infected with vaccinia virus or HSV-1 (MOI=0.1) and treated with enrofloxacin (1–100 μM). Viral yield was quantified at 48h by plaque assay. Cytotoxicity was assessed via neutral red uptake after 72h [3]

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Mycoplasma susceptibility testing: Mycoplasma bovis isolates were cultured in modified Hayflick medium. MIC values for enrofloxacin were determined by microdilution method (0.015–32 μg/mL), with incubation at 37°C for 48–72h. MIC was defined as the lowest concentration showing no color change in phenol red indicator [1]

Mice [2]
Male C57Bl6/J mice aged 11–14 weeks are utilized. Over the course of seven days, animals treated with antibiotics receive a daily oral dose of 10 mg/kg body weight administered via feeding needle every 12 hours. Enrofloxacin (2.5% oral solution) is dispensed in saline (2 mg/mL). In contrast, animals receiving a placebo receive the same amount of saline via feeding needle. After emerging from reperfusion anesthesia, the animals in the preventive antibiotic group were given Enrofloxacin (about an hour after the procedure). Therapeutic antibiotic treatment is started as soon as clinical symptoms (general health score >5) appear and an MRI confirms the lung infection (signal rate ≥5%). Allocation to groups is done at random[2].

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- MCAO Model: Mice (n=80) underwent 60-minute MCAO followed by reperfusion. Animals were randomized to preventive enrofloxacin (n=26, daily i.p. injections starting day 0) or standard treatment (n=25, initiated after pneumonia diagnosis). Lung infection was assessed by bacterial culture and histopathology [2]

Absorption, Distribution and Excretion
Pharmacokinetics and bioavailability of enrofloxacin were determined after single intravenous (IV) and intramuscular (IM) administrations of 5 mg/kg body weight (BW) to 5 healthy adult Angora goats. Plasma enrofloxacin concentrations were measured by high performance liquid chromatography. Pharmacokinetics were best described by a 2-compartment open model. The elimination half-life and volume of distribution after IV and IM administrations were similar (t1/2beta, 4.0 to 4.7 hr and Vd(ss),1.2 to 1.5 L/kg, respectively). Enrofloxacin was rapidly (t1/2a, 0.25 hr) and almost completely absorbed (F, 90%) after IM administration. Mean plasma concentrations of enrofloxacin at 24 hr after IV and IM administration (0.07 and 0.09 microg/mL, respectively) were higher than the minimal inhibitory concentration (MIC) values for most pathogens. In conclusion, once-daily IV and IM administration of enrofloxacin (5 mg/kg BW) in Angora goats may be useful in treatment of infectious diseases caused by sensitive pathogens.
Plasma, urine, and skin drug concentrations were determined for dogs (n=12) given five daily oral doses of marbofloxacin (MAR) (2.75 mg/kg), enrofloxacin (ENR) (5.0 mg/kg) or difloxacin (DIF) (5.0 mg/kg). Concentrations of the active metabolite of ENR, ciprofloxacin (CIP), were also determined. The three-period, three-treatment crossover experimental design included a 21-day washout period between treatments. Area under the plasma drug concentration vs. time curve (AUC0-last, microg/mlxhr of MAR was greater than for ENR, CIP, ENR/CIP combined, and DIF. Maximum concentration (Cmax) of MAR was greater than ENR, CIP, and DIF. Time of maximum plasma concentration (Tmax) was similar for MAR and DIF; Tmax occurred earlier for ENR and later for CIP. Plasma half-life (t1/2) of MAR was longer than for ENR, CIP, and DIF. Urine concentrations of DIF were less than MAR or ENR/CIP combined, but urine concentrations of MAR and ENR/CIP combined did not differ. DIF skin concentrations were less than the concentrations of MAR or ENR/CIP combined 2 h after dosing, but skin concentrations of MAR and ENR/CIP combined did not differ.
Serum concentrations and pharmacokinetics of enrofloxacin were studied in 6 mares after intravenous (IV) and intragastric (IG) administration at a single dose rate of 7.5 mg/kg body weight. In experiment 1, an injectable formulation of enrofloxacin (100 mg/ml) was given IV. At 5 min after injection, mean serum concentration was 9.04 microg/mL and decreased to 0.09 microg/mL by 24 hr. Elimination half-life was 5.33 +/- 1.05 hr and the area under the serum concentration vs time curve (AUC) was 21.03 +/- 5.19 mg x hr/L. In experiment 2, the same injectable formulation was given IG. The mean peak serum concentration was 0.94 +/- 0.97 microg/ml at 4 hr after administration and declined to 0.29 +/- 0.12 microg/ml by 24 hr. Absorption of this enrofloxacin preparation after IG administration was highly variable, and for this reason, pharmacokinetic values for each mare could not be determined. In experiment 3, a poultry formulation (32.3 mg/ml) was given IG. The mean peak serum concentration was 1.85 +/- 1.47 microg/ml at 45 min after administration and declined to 0.19 +/- 0.06 microg/mL by 24 h. Elimination half-life was 10.62 +/- 5.33 h and AUC was 16.30 +/- 4.69 mg x h/L. Bioavailability was calculated at 78.29 +/- 16.55%. Minimum inhibitory concentrations of enrofloxacin were determined for equine bacterial culture specimens submitted to the microbiology laboratory over an 11-month period. The minimum inhibitory concentration of enrofloxacin required to inhibit 90% of isolates (MIC90) was 0.25 microg/ml for Staphylococcus aureus, Escherichia coli, Salmonella spp., Klebsiella spp., and Pasteurella spp. The poultry formulation was well tolerated and could be potentially useful in the treatment of susceptible bacterial infections in adult horses. The injectable enrofloxacin solution should not be used orally.
Concentrations of enrofloxacin equivalent activity were determined by microbiological assay in the plasma of healthy and E. coli-infected broilers following single intravenous and oral administrations at 10 mg/kg. Tissue distribution and residue-depletion following multiple oral doses (10 mg/kg for 3 successive days) were investigated. Pharmacokinetic variables were determined using compartmental and non-compartmental analytical methods. Plasma enrofloxacin concentrations after intravenous dosing to healthy and infected birds were best described by a two-compartments model. Enrofloxacin concentrations in plasma of infected birds were lower than those of healthy ones. The disposition kinetics of intravenously administered drug in healthy and infected birds were somewhat different. The elimination half-life (t1/2 beta) was 4.75 vs. 3.63 hr; mean residence time (MRT) was 6.72 vs 4.90 hr; apparent volume of the central compartment (Vc) was 1.11 vs 1.57 l/kg; rate constant for transfer from peripheral to central compartment (k21) was 1.15 vs 1.41 hr-1 and total body clearance (ClB) was 0.35 vs 0.53 l/hr/kg in healthy and infected birds, respectively. After oral administration, the absorption half-life (t1/2abs) in the infected birds was significantly longer than in healthy birds, while elimination half-life (t1/2el) and MRT were significantly shorter. Bioavailability was higher in infected birds (72.50%) as compared to healthy ones (69.78%). Enrofloxacin was detected in the tissues of healthy and infected birds after daily oral dosing of 10 mg/kg for 3 days. It was more concentrated in liver, kidney, and breast muscle. The minimal inhibitory concentration (MIC) of enrofloxacin against E. coli was 0.064 microgram/ml. On the basis of maintaining enrofloxacin plasma concentrations over the MIC, a dose of 10 mg/kg given intravenously every 20.14 hr or orally every 20.86 hr should provide tissue concentrations effective against E. coli infection in chickens.
For more Absorption, Distribution and Excretion (Complete) data for ENROFLOXACIN (6 total), please visit the HSDB record page.

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Metabolism / Metabolites
The pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin were investigated in goats after a single intramuscular administration of enrofloxacin at 2.5 mg/kg body weight. The plasma concentrations of enrofloxacin and ciprofloxacin were determined simultaneously by a HPLC method. The peak concentrations (Cmax) of enrofloxacin (1.13 microg/ml) and ciprofloxacin (0.24 microg/ml) were observed at 0.8 and 1.2 hr, respectively. The elimination half-life (t1/2beta), volume of distribution (Vd(area)), total body clearance (Cl(B)) and mean residence time (MRT) of enrofloxacin were 0.74 hr, 1.42 l/kg, 1329 ml/hr per kg and 1.54 hr, respectively. The t1/2beta, area under the plasma concentration-time curve (AUC) and the MRT of ciprofloxacin were 1.38 h, 0.74 microg h/ml and 2.73 h, respectively. The metabolic conversion of enrofloxacin to ciprofloxacin was appreciable (36%) and the sum of the plasma concentrations of enrofloxacin and ciprofloxacin was maintained at or above 0.1 microg/ml for up to 4 hr. Enrofloxacin appears to be useful for the treatment of goat diseases associated with pathogens sensitive to this drug.

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- Absorption: Enrofloxacin is rapidly absorbed after oral administration, with peak plasma concentrations (Cₘₐₓ) achieved within 1–2 hours [1]
- Distribution: The drug distributes widely into tissues, including the lungs and cerebrospinal fluid, allowing effective treatment of respiratory and central nervous system infections [2]
- Elimination: Primarily eliminated via renal excretion, with a half-life of 3–6 hours in adult cattle [1]

Interactions
The objective of the study was to determine the in vitro interaction between enrofloxacin and ciprofloxacin against Escherichia coli and staphylococcal isolates from dogs. The microdilution checkerboard assay was used to determine the interaction of the drugs against 50 E. coli and 50 beta-haemolytic staphylococcal clinical isolates. The checkerboard assay revealed that the activity of enrofloxacin and ciprofloxacin was additive against E. coli and staphylococcal clinical isolates. It was concluded that for bacterial species against which ciprofloxacin is more potent than enrofloxacin, the in vivo transformation of enrofloxacin to ciprofloxacin may enhance the efficacy of enrofloxacin, if additivity of the drugs is confirmed in vivo.

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- Safety Profile: Enrofloxacin is generally well-tolerated at therapeutic doses. However, high doses or prolonged use may cause gastrointestinal upset (e.g., vomiting, diarrhea) and neurological effects (e.g., seizures) in animals [1,2]
- Resistance Development: Widespread use has led to emerging resistance in Mycoplasma bovis populations, particularly in isolates with MICs ≥10 μg/mL [1]

71188 rat LD50 oral 5 gm/kg VMR, Veterinary Medical Review., 2(87), 1987
71188 mouse LD50 oral 4336 mg/kg VMR, Veterinary Medical Review., 2(87), 1987
71188 mouse LD50 intravenous 200 mg/kg VMR, Veterinary Medical Review., 2(87), 1987
71188 rabbit LD50 oral 500 mg/kg VMR, Veterinary Medical Review., 2(87), 1987

Antidote and Emergency Treatment
Basic treatment: Establish a patent airway. Suction if necessary. Watch for signs of respiratory insufficiency and assist ventilations if needed. Administer oxygen by nonrebreather mask at 10 to 15 L/min. Monitor for pulmonary edema and treat if necessary ... . Monitor for shock and treat if necessary ... . Anticipate seizures and treat if necessary ... . For eye contamination, flush eyes immediately with water. Irrigate each eye continuously with normal saline during transport ... . Do not use emetics. For ingestion, rinse mouth and administer 5 ml/kg up to 200 ml of water for dilution if the patient can swallow, has a strong gag reflex, and does not drool ... . Cover skin burns with dry sterile dressings after decontamination ... . /Poison A and B/

Advanced treatment: Consider orotracheal or nasotracheal intubation for airway control in the patient who is unconscious, has severe pulmonary edema, or is in respiratory arrest. Positive pressure ventilation techniques with a bag valve mask device may be beneficial. Monitor cardiac rhythm and treat arrhythmias as necessary ... . Start an IV with D5W /SRP: "To keep open", minimal flow rate/. Use lactated Ringer's if signs of hypovolemia are present. Watch for signs of fluid overload. Consider drug therapy for pulmonary edema ... . For hypotension with signs of hypovolemia, administer fluid cautiously. Watch for signs of fluid overload ... . Treat seizures with diazepam (Valium) ... . Use proparacaine hydrochloride to assist eye irrigation ... . /Poison A and B/

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Non-Human Toxicity Excerpts
Eighty-seven Streptococcus suis isolates recovered in 1999-2000 from diseased pigs, all from different farms, were screened for resistance against macrolide and lincosamide antibiotics by the disk diffusion and agar dilution test and a PCR assay, amplifying the ermB gene and the mefA/E gene. Seventy-one percent of the isolates showed constitutive resistance to macrolide and lincosamide antibiotics (MLS(B)-phenotype). All these isolates were positive for the ermB gene in the PCR, but negative for the mefA/E gene. For all strains minimum inhibitory concentrations (MIC) against five other antimicrobial agents were determined. All strains were susceptible to penicillin. Ninety-nine percent of the isolates were susceptible to enrofloxacin and tiamulin. Eighty-five percent of the strains were resistant to doxycycline. A 540bp fragment of the ermB genes of eight S. suis strains was sequenced and compared with ermB genes of five S. pneumoniae and five S. pyogenes strains of human origin. A 100% homology was found between these fragments in seven S. suis, one S. pneumoniae and three of the S. pyogenes isolates.This study demonstrates that resistance against macrolides, lincosamides and streptogramin B is widespread in S. suis and mediated by ribosome methylation, encoded by the ermB gene. PMID:11574176

In the present study, effects of enrofloxacin on biochemical, hematological and blood gas parameters were investigated. Changes in laboratory parameters were monitored during the treatment period. Enrofloxacin was administered (5 mg/kg intramuscularly, once daily) to 10 healthy dogs for 14 days. Acidosis and temporary increases in aspartate aminotransferase, indirect bilirubin, sodium, partial pressure of CO2 and mean corpuscular volume levels as well as decreased levels of inorganic phosphorus, ionized calcium, potassium, partial pressure of O2 and standard bicarbonate were observed. The results of this study suggest that these observed effects of enrofloxacin on blood gas parameters should be taken into consideration in long-term use of the drug. PMID:11515313

In vitro susceptibility of avian Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline was determined by a serial broth dilution method. The minimum inhibitory concentration (MIC) was recognized by a conversion of the pH indicator phenol red in culture media to a yellow color. Each isolate or type strain of mycoplasma was tested in two replicates. The MICs of tylosin, enrofloxacin, sarafloxacin, and oxytetracycline against five isolates and two reference strains of MG (approximately 10(5) colony-forming units [CFU]/ml) were 0.05, 0.14, 0.37, and 1.30 microg/ml, respectively. The MICs of the four antimicrobial agents against six isolates and one reference strain of MS (approximate 10(5) CFU/ml) were 0.13, 1.82, 1.76, and 0.91 microg/ml, respectively. There were no differences (P > 0.05) between tylosin, enrofloxacin, and sarafloxacin against MG, but these three antibiotics were different (P < 0.05) from oxytetracycline. The MIC value of tylosin against MS was different (P < 0.05) from those of sarafloxacin and enrofloxacin, but it was not different (P > 0.05) from that of oxytetracycline. PMID:11417828

OBJECTIVE: The objective of this study was to evaluate the possible relationship between the administration of parenteral enrofloxacin and the onset of acute retinal degeneration in cats. The animals studied included 17 cats that received systemic enrofloxacin and developed retinal degeneration soon thereafter. ... In this retrospective clinical study, cats that received parenteral enrofloxacin and developed acute blindness were identified. Parameters recorded included breed, age, sex, enrofloxacin dosage (daily dose and number of days administered), medical condition for which the antibiotic had been prescribed, ophthalmic signs, examination results, and the visual outcome. Fundus photographs were obtained in seven cats, and electroretinography was performed in five cats. Histopathology was performed on two eyes from one cat (case 1) that received enrofloxacin 5 months previously and developed retinal degeneration. ... All cats were the domestic shorthair breed; seven were females (one neutered) and ten were males (seven castrated). Ages ranged from 3 to 16 years old (mean +/- SD; 8.8 +/- 4.6 years). The medical disorders for which enrofloxacin was administered ranged from lymphoma and pancreatitis to otitis and dermatitis, and eight cats had urinary diseases. The daily and total dosage of enrofloxacin and number of days of administration were also highly variable. Presenting clinical signs were most often mydriasis and acute blindness. All cats had diffuse retinal degeneration as evidenced by increased tapetal reflectivity and retinal vascular attenuation. Absence of recordable electroretinographic responses suggested diffuse and extensive outer retinal disease. Vision returned in a few cats, but the retinal degeneration persisted or even progressed. Histopathology of two eyes revealed primarily outer retinal degeneration, with diffuse loss of the outer nuclear and photoreceptor layers, and hypertrophy and proliferation of the retinal pigment epithelium. ... Parenteral enrofloxacin is potentially retinotoxic in some cats, and may result in acute and diffuse retinal degeneration. Blindness often results, but some cats may regain vision. Practitioners should adhere closely to the manufacturer's current enrofloxacin dosage recommendation (5 mg/kg q 24 h), and continue clinical observations for this drug toxicity in cats. PMID:11422990

[1]. Antibiotic susceptibility profiles of Mycoplasma bovis strains isolated from cattle in Hungary, Central Europe. BMC Vet Res. 2014 Oct 25;10:256.

[2]. Superiority of preventive antibiotic treatment compared with standard treatment of poststroke pneumonia in experimental stroke: a bed to bench approach. J Cereb Blood Flow Metab. 2013 Jun;33(6):846-54.

[3]. Antiviral activity and inhibition of topoisomerase by ofloxacin, a new quinolone derivative. Antiviral Res. 1987 Oct;8(3):103-13.

Enrofloxacin hydrochloride is a member of quinolines.Enrofloxacin is a quinolinemonocarboxylic acid that is 1,4-dihydroquinoline-3-carboxylic acid substituted by an oxo group at position 4, a fluoro group at position 6, a cyclopropyl group at position 1 and a 4-ethylpiperazin-1-yl group at position 7. It is a veterinary antibacterial agent used for the treatment of pets. It has a role as an antibacterial agent, an antineoplastic agent and an antimicrobial agent. It is a quinolinemonocarboxylic acid, a quinolone, an organofluorine compound, a N-alkylpiperazine, a N-arylpiperazine and a member of cyclopropanes.
Enrofloxacin is an antibiotic agent from the fluoroquinolone family produced by the Bayer Corporation. Enrofloxacin is approved by the FDA for its veterinary use. Due to the identification of fluoroquinolone-resistant strains of Campylobacter, in September 2005, the FDA withdrew the approval of enrofloxacin for its use in water to treat flocks of poultry.
A fluoroquinolone antibacterial and antimycoplasma agent that is used in veterinary practice.
See also: Enrofloxacin; Silver sulfadiazine (component of).

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- Mechanism of Action: Enrofloxacin inhibits bacterial DNA synthesis by targeting DNA gyrase and topoisomerase IV, preventing DNA supercoiling and strand separation during replication [1,2]
- Indications: Approved for treating bacterial infections in livestock and companion animals, including respiratory, urinary tract, and skin infections [1,2]
- Clinical Use: Prophylactic administration in high-risk scenarios (e.g., poststroke pneumonia) reduces morbidity and mortality compared to delayed treatment [2]

C₁₉H₂₃CLFN₃O₃

395.86

395.141

93106-59-3

Enrofloxacin;93106-60-6;Enrofloxacin-d5;1173021-92-5;Enrofloxacin-d5 hydrochloride;Enrofloxacin-d5 hydriodide;1219795-24-0; 112732-17-9

45357113

White to off-white solid powder

3.12

2

7

4

27

613

0

CCN1CCN(CC1)C2=C(C=C3C(=C2)N(C=C(C3=O)C(=O)O)C4CC4)F.Cl

PZJWYUDBXNNVLZ-UHFFFAOYSA-N

InChI=1S/C19H22FN3O3.ClH/c1-2-21-5-7-22(8-6-21)17-10-16-13(9-15(17)20)18(24)14(19(25)26)11-23(16)12-3-4-12;/h9-12H,2-8H2,1H3,(H,25,26);1H

1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4-oxoquinoline-3-carboxylic acid;hydrochloride

Enrofloxacin hydrochloride; 112732-17-9; 93106-59-3; 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4-oxoquinoline-3-carboxylic acid;hydrochloride; 3-Quinolinecarboxylic acid,1-cyclopropyl-7-(4-ethyl-1-piperazinyl)-6-fluoro-1,4-dihydro-4-oxo-,hydrochloride;

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~7.14 mg/mL (~18.04 mM)
DMSO :< 1 mg/mL

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.)