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

- 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].

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
The pharmacokinetics and bioavailability of enrofloxacin were determined in five healthy adult Angora goats after single intravenous (IV) and intramuscular (IM) injections of 5 mg/kg body weight (BW). Plasma enrofloxacin concentrations were determined using high-performance liquid chromatography (HPLC). The pharmacokinetics best conformed to a two-compartment open model. The elimination half-life and volume of distribution were similar after IV and IM injections (t1/2β were 4.0–4.7 hours and Vd(ss) were 1.2–1.5 L/kg, respectively). Following IM injection, enrofloxacin was rapidly absorbed (t1/2a was 0.25 hours) and almost completely absorbed (F was 90%). Twenty-four hours after both IV and IM injections, the mean plasma concentrations (0.07 and 0.09 μg/mL, respectively) were higher than the minimum inhibitory concentrations (MICs) for most pathogens. In summary, once-daily intravenous and intramuscular administration of enrofloxacin (5 mg/kg body weight) in Angola goats may be helpful in treating infectious diseases caused by susceptible pathogens. Plasma, urine, and skin concentrations were determined in 12 dogs after five oral administrations of marbofloxacin (MAR) (2.75 mg/kg), enrofloxacin (ENR) (5.0 mg/kg), or desflufloxacin (DIF) (5.0 mg/kg). The concentration of ciprofloxacin (CIP), the active metabolite of enrofloxacin, was also determined. This three-cycle, three-treatment crossover design included a 21-day washout period between treatments. The area under the plasma drug concentration-time curve (AUC0-last, μg/ml·hr) for MAR was greater than that for ENR, CIP, the combination of ENR/CIP, and DIF. The maximum concentration (Cmax) of MAR was greater than that of ENR, CIP, and DIF. The time to maximum plasma concentration (Tmax) was similar for MAR and DIF; Tmax appeared earlier for ENR and later for CIP. The plasma half-life (t1/2) of MAR was longer than that of ENR, CIP, and DIF. Urinary concentrations of DIF were lower than those of MAR or the combined use of ENR/CIP, but there was no difference in urinary concentrations between MAR and the combined use of ENR/CIP. Two hours after administration, skin concentrations of DIF were lower than those of MAR or the combined use of ENR/CIP, but there was no difference in skin concentrations between MAR and the combined use of ENR/CIP. This study investigated serum concentrations and pharmacokinetics of enrofloxacin after intravenous injection in six mares. Two routes of administration were used: intravenous injection (IV) and intragastric instillation (IG), with a single dose of 7.5 mg/kg body weight. In Experiment 1, enrofloxacin injection (100 mg/ml) was administered intravenously. Five minutes after injection, the mean serum concentration was 9.04 μg/mL, decreasing to 0.09 μg/mL after 24 hours. The elimination half-life was 5.33 ± 1.05 hours, and the area under the serum concentration-time curve (AUC) was 21.03 ± 5.19 mg·hr/L. In Experiment 2, the same injection solution was administered intragastricly. Four hours after administration, the mean peak serum concentration was 0.94 ± 0.97 μg/ml, decreasing to 0.29 ± 0.12 μg/ml after 24 hours. Enrofloxacin absorption after intragastric administration varied considerably; therefore, pharmacokinetic parameters for each mare could not be determined. In Experiment 3, mares were intravenously injected with a poultry formulation (32.3 mg/ml). Forty-five minutes after administration, the mean peak serum concentration was 1.85 ± 1.47 μg/ml, decreasing to 0.19 ± 0.06 μg/ml after 24 hours. The elimination half-life was 10.62 ± 5.33 hours, and the AUC was 16.30 ± 4.69 mg·h/L. The bioavailability was calculated to be 78.29 ± 16.55%. The minimum inhibitory concentration (MIC) of enrofloxacin was determined in equine bacterial cultures submitted to the microbiology laboratory within 11 months. The MIC90 for inhibiting 90% of isolates was 0.25 μg/ml, suitable for Staphylococcus aureus and Escherichia coli, Escherichia coli, Salmonella spp., Klebsiella spp., and Pasteurella spp. The poultry formulation was well tolerated and may have potential use in treating susceptible bacterial infections in adult horses. Enrofloxacin solution for injection should not be administered orally. The concentrations of enrofloxacin equivalent activity in plasma were determined using microbiological methods in healthy broilers and broilers infected with Escherichia coli after a single intravenous injection and oral administration of 10 mg/kg enrofloxacin. Tissue distribution and residual clearance after multiple oral administrations (10 mg/kg for 3 consecutive days) were investigated. Pharmacokinetic parameters were determined using both compartmental and non-compartmental model analyses. Plasma concentrations of enrofloxacin after intravenous injection in healthy and infected chickens best conformed to the two-compartment model. Enrofloxacin concentrations in the plasma of infected chickens were lower than those in healthy chickens. Drug distribution kinetics after intravenous injection showed slightly different responses in healthy and infected poultry. The elimination half-life (t_1/2β) in healthy and infected poultry were 4.75 h and 3.63 h, respectively; the mean residence time (MRT) was 6.72 h and 4.90 h, respectively; the apparent volume of the central compartment (V_c) was 1.11 L/kg and 1.57 L/kg, respectively; the transport rate constant from the peripheral compartment to the central compartment (k₂₁) was 1.15 h^-1 and 1.41 h^-1, respectively; and the total clearance (Cl_B) was 0.35 L/h/kg and 0.53 L/h/kg, respectively. Following oral administration, the absorption half-life (t_1/2abs) of infected poultry was significantly longer than that of healthy poultry, while the elimination half-life (t_1/2el) and mean residence time (MRT) were significantly shorter. Bioavailability in infected poultry (72.50%) was higher than in healthy poultry (69.78%). Enrofloxacin was detected in tissues of both healthy and infected poultry. The daily oral dose was 10 mg/kg for 3 consecutive days. High drug concentrations were observed in the liver, kidneys, and pectoral muscles. The minimum inhibitory concentration (MIC) of enrofloxacin against E. coli was 0.064 μg/ml. Based on maintaining enrofloxacin plasma concentrations above the MIC, an intravenous injection every 20.14 hours or oral administration every 20.86 hours of 10 mg/kg should achieve effective drug concentrations in chicken tissues against E. coli infection.
For more complete data on absorption, distribution, and excretion of enrofloxacin (6 items in total), please visit the HSDB record page.

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Metabolism/Metabolites
In goats, the pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin were studied following a single intramuscular injection of 2.5 mg/kg body weight of enrofloxacin. Plasma concentrations of enrofloxacin and ciprofloxacin were determined simultaneously by high-performance liquid chromatography. Peak concentrations (Cmax) of enrofloxacin (1.13 μg/ml) and ciprofloxacin (0.24 μg/ml) occurred at 0.8 h and 1.2 h, respectively. The elimination half-life (t1/2β), volume of distribution (Vd(area)), systemic clearance (Cl(B)), and mean residence time (MRT) of enrofloxacin were 0.74 h, 1.42 L/kg, 1329 mL/h/kg, and 1.54 h, respectively. The t1/2β, area under the plasma concentration-time curve (AUC), and MRT of ciprofloxacin were 1.38 h, 0.74 μg·h/mL, and 2.73 h, respectively. The metabolic conversion of enrofloxacin to ciprofloxacin was significant (36%), and the sum of the plasma concentrations of enrofloxacin and ciprofloxacin remained at 0.1 μg/mL or higher for up to 4 hours. Enrofloxacin appears to be useful for treating goat diseases caused by pathogens susceptible to the drug.

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- Absorption: Enrofloxacin is rapidly absorbed after oral administration, with peak plasma concentrations (Cₘₐₓ) reached within 1–2 hours [1]
- Distribution: The drug is widely distributed in tissues including the lungs and cerebrospinal fluid and is effective in treating respiratory and central nervous system infections [2]
- Elimination: It is mainly excreted by the kidneys, with a half-life of 3–6 hours in adult cattle [1]

Interactions
This study aimed to determine the in vitro interactions of enrofloxacin and ciprofloxacin against canine Escherichia coli and Staphylococcus isolates. The interactions of the two drugs against 50 clinical isolates of E. coli and 50 clinical isolates of β-hemolytic Staphylococcus were determined using the microdilution checkerboard method. Checkerboard results showed an additive effect of the activities of enrofloxacin and ciprofloxacin against the clinical isolates of E. coli and Staphylococcus. The conclusion is that for bacteria where ciprofloxacin is more effective than enrofloxacin, if an additive effect is confirmed in vivo, the conversion of enrofloxacin to ciprofloxacin in vivo may enhance the efficacy of enrofloxacin. Safety Profile: Enrofloxacin is generally well tolerated at therapeutic doses. However, high doses or prolonged use may lead to gastrointestinal discomfort (e.g., vomiting, diarrhea) and neurological symptoms (e.g., seizures) in animals [1,2] - Development of resistance: Widespread use has led to the emergence of resistance in bovine mycoplasma populations, particularly in strains with MIC ≥ 10 μg/mL [1] 71188trattLD50toralt5 gm/kgtVMR, Veterinary Medicine Review, 2(87), 1987 71188tmousetLD50toralt4336 mg/kgtVMR, Veterinary Medicine Review, 2(87), 1987 71188tmousetLD50tintravenoust200 mg/kgtVMR, Veterinary Medicine Review, 2(87), 1987 71188 rabbittLD50toralt500 mg/kgtVMR, Veterinary Medicine Review, 2(87), 1987

Antidotes and Emergency Treatment
Basic Treatment: Maintain a clear airway. Suction if necessary. Observe for signs of respiratory failure and provide assisted ventilation if necessary. Administer oxygen via a non-invasive ventilation mask at a flow rate of 10 to 15 liters/minute. Monitor for pulmonary edema and treat if necessary… Monitor for shock and treat if necessary… Anticipate seizures and treat if necessary… If eyes are contaminated, flush with water immediately. During transport, continuously flush each eye with saline… Do not use emetics. If swallowed, rinse mouth; if the patient is able to swallow, has a strong gag reflex, and does not drool, dilute with 5 ml/kg to 200 ml of water… After disinfection, cover burns with a dry, sterile dressing…/Class A and Class B Poisons/
Advanced Treatment: For patients with impaired consciousness, severe pulmonary edema, or respiratory arrest, consider oropharyngeal or nasopharyngeal endotracheal intubation to control the airway. Positive pressure ventilation using a bag-valve-mask may be effective. Monitor cardiac rhythm and treat arrhythmias if necessary…. Establish intravenous access using 5% glucose solution (SRP: maintain patency of the intravenous access at the minimum flow rate). If signs of hypovolemia appear, use lactated Ringer's solution. Watch for signs of fluid overload. Consider medical treatment for pulmonary edema…. For hypotension with signs of hypovolemia, administer fluids with caution. Watch for signs of fluid overload…. Use diazepam (Valium) to treat seizures…. Use promecaine hydrochloride to assist eye irrigation…. /Toxins A and B/

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Excerpt of Non-Human Toxicity
Between 1999 and 2000, 87 strains of Streptococcus suis were isolated from diseased pigs in different farms. Resistance to macrolides and lincosamides was screened using disk diffusion, agar dilution, and PCR (amplification of the ermB and mefA/E genes). 71% of the isolates showed constitutive resistance to macrolides and lincosamides (MLS(B) phenotype). PCR detection of the ermB gene was positive for all isolates, while the mefA/E gene was negative. The minimum inhibitory concentrations (MICs) of five other antimicrobial agents were determined for all strains. All strains were sensitive to penicillin. 99% of the isolates were sensitive to enrofloxacin and tiamulin. 85% of the strains were resistant to doxycycline. The 540 bp fragment of the ermB gene from eight Streptococcus suis isolates was sequenced and compared with the ermB genes from five human Streptococcus pneumoniae isolates and five human Streptococcus pyogenes isolates. The results showed that the ermB gene fragments from seven Streptococcus suis isolates, one Streptococcus pneumoniae isolate, and three Streptococcus pyogenes isolates shared 100% homology. This study indicates that Streptococcus suis commonly exhibits resistance to macrolides, lincosamides, and streptomycin B antibiotics, and that this resistance is mediated by ribosomal methylation encoded by the ermB gene. PMID:11574176
This study investigated the effects of enrofloxacin on biochemical, hematological, and blood gas parameters. Changes in laboratory parameters were monitored during treatment. Ten healthy dogs received intramuscular injections of enrofloxacin (5 mg/kg, once daily) for 14 consecutive days. Acidosis was observed, along with transient increases in aspartate aminotransferase, indirect bilirubin, sodium, partial pressure of carbon dioxide, and mean corpuscular volume, while levels of inorganic phosphorus, ionic calcium, potassium, partial pressure of oxygen, and standard bicarbonate decreased. These results suggest that these effects on blood gas parameters should be considered during long-term enrofloxacin use. PMID:11515313

The in vitro susceptibility of avian mycoplasma gallisepticum (MG) and mycoplasma synoviae (MS) to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline was determined using a serial broth dilution method. The minimum inhibitory concentration (MIC) was determined by the change of phenol red to yellow in the pH indicator of the culture medium. Each mycoplasma isolate or standard strain was tested twice. The MICs of tylosin, enrofloxacin, sarafloxacin, and oxytetracycline against 5 MG isolates and 2 reference strains (approximately 10⁵ CFU/ml) were 0.05, 0.14, 0.37, and 1.30 μg/ml, respectively. The MICs of these four antimicrobial agents against 6 MS isolates and 1 reference strain (approximately 10⁵ CFU/ml) were 0.13, 1.82, 1.76, and 0.91 μg/ml, respectively. There was no significant difference in antimicrobial activity between tylosin, enrofloxacin, and sarafloxacin against MG (P > 0.05), but a significant difference compared to oxytetracycline (P < 0.05). The MIC of tylosin against MS was significantly different from that of sarafloxacin and enrofloxacin (P < 0.05), but not significantly different from that of oxytetracycline (P > 0.05). PMID:11417828

Objective: This study aimed to evaluate the possible relationship between parenteral enrofloxacin administration and the occurrence of acute retinal degeneration in cats. The study included 17 cats that developed retinal degeneration shortly after receiving systemic enrofloxacin treatment. …In this retrospective clinical study, we screened cats that developed acute blindness after receiving parenteral enrofloxacin treatment. Recorded parameters included breed, age, sex, enrofloxacin dosage (daily dose and number of days of administration), indication for the antibiotic, ocular symptoms, examination results, and visual prognosis. Fundus photography was performed on 7 cats, and electroretinography was performed on 5 cats. Histopathological examination was performed on both eyes of one cat (Case 1) that developed retinal degeneration 5 months prior after enrofloxacin treatment. …All cats were domestic shorthaired cats; 7 were female (1 spayed) and 10 were male (7 neutered). The age range was 3 to 16 years (mean ± standard deviation: 8.8 ± 4.6 years). Diseases treated with enrofloxacin included lymphoma, pancreatitis, otitis, and dermatitis; eight cats also had urinary tract infections. There was considerable variation in daily dose, total dose, and number of days of treatment with enrofloxacin. The most common clinical manifestations were mydriasis and acute blindness. All cats developed diffuse retinal degeneration, characterized by thickening of the retinal reflective layer and thinning of retinal vessels. Electroretinography did not record diffuse, widespread outer retinal lesions. A few cats experienced visual recovery, but retinal degeneration persisted or even worsened. Histopathological examination of both eyes revealed predominantly outer retinal degeneration, with diffuse loss of the outer nuclear layer and photoreceptor layer, as well as hypertrophy and hyperplasia of the retinal pigment epithelium. …Injectable enrofloxacin may be retinal toxic in some cats and may cause acute diffuse retinal degeneration. Blindness is usually permanent, but some cats may regain vision. Clinicians should strictly adhere to the manufacturer's current recommended dosage of enrofloxacin (5 mg/kg, every 24 hours) and continue clinical observation of cats for drug toxicity. 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 is a quinoline monocarboxylic acid with the structure 1,4-dihydroquinoline-3-carboxylic acid, substituted with a carbonyl group at position 4, a fluorine group at position 6, a cyclopropyl group at position 1, and a 4-ethylpiperazine-1-yl group at position 7. It is a veterinary antibacterial drug used to treat pets. Enrofloxacin has antibacterial, antitumor, and antimicrobial activities. It is a quinoline monocarboxylic acid, a quinolone compound, an organofluorine compound, an N-alkylpiperazine compound, an N-arylpiperazine compound, and also belongs to the cyclopropane class of compounds. Enrofloxacin is a fluoroquinolone antibiotic manufactured by Bayer AG. Enrofloxacin was approved by the U.S. Food and Drug Administration (FDA) for veterinary use. However, in September 2005, the FDA revoked its approval for the treatment of poultry in drinking water due to the discovery of Campylobacter strains resistant to fluoroquinolones. It is a fluoroquinolone antibacterial and antimycoplasma drug used in veterinary practice. See also: Enrofloxacin; Silver sulfadiazine (component). - Mechanism of action: Enrofloxacin inhibits bacterial DNA synthesis by targeting DNA gyrase and topoisomerase IV, thereby preventing DNA supercoiling and strand separation during replication [1,2]. - Indications: Approved for the treatment of bacterial infections in livestock and companion animals, including respiratory, urinary, and skin infections [1,2]. - Clinical use: Prophylactic administration in high-risk situations (e.g., post-stroke pneumonia) reduces morbidity and mortality compared to delayed treatment [2].

C19H22FN3O3

359.3947

359.164

C, 63.50; H, 6.17; F, 5.29; N, 11.69; O, 13.36

93106-60-6

Enrofloxacin monohydrochloride;93106-59-3;Enrofloxacin-d5;1173021-92-5

71188

Light yellow to yellow solid powder

1.4±0.1 g/cm3

560.5±50.0 °C at 760 mmHg

225 °C

292.8±30.1 °C

0.0±1.6 mmHg at 25°C

1.634

1.88

1

7

4

26

613

0

O=C(C1C(=O)C2C(=CC(N3CCN(CC)CC3)=C(C=2)F)N(C2CC2)C=1)O

SPFYMRJSYKOXGV-UHFFFAOYSA-N

InChI=1S/C19H22FN3O3/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)

3-Quinolinecarboxylic acid, 1,4-dihydro-1-cyclopropyl-7-(4-ethyl-1-piperazinyl)-6-fluoro-4-oxo-, hydrochloride

Baytril; Enrofloxacine; CFPQ; 93106-60-6; Baytril; Enrofloxacine; Enrofloxacino; Enrofloxacinum; BAY VP 2674; endrofloxicin; Bay-Vp-2674; BAY-Vp2674; PD 160788; BAY-Vp2674; PD160788; BAY-Vp 2674; PD-160788; endrofloxicin.

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)

4-Methylpyridine : ~30 mg/mL
H2O : ~1 mg/mL (~2.78 mM)
DMSO : 1~10 mg/mL ( 2.78~27.82 mM )

Solubility in Formulation 1: ≥ 1 mg/mL (2.78 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 1 mg/mL (2.78 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 1 mg/mL (2.78 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 1 mg/mL (2.78 mM)

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