Gamithromycin

Background:Gamithromycin is active in vitro against the bacterial agents most commonly associated with bronchopneumonia in older foals. However, the clinical efficacy and safety of this drug have not been investigated.Hypothesis:Gamithromycin is effective for the treatment of bronchopneumonia in foals.Animals:One hundred and twenty-one foals on a farm endemic for infections caused by Rhodococcus equi.Methods:In a controlled, randomized, and double blinded clinical trial, foals with ultrasonographic evidence of pulmonary abscesses (abscess score 8.0-20 cm) were randomly allocated in 3 treatment groups: (1) gamithromycin IM q7 days (n = 40); (2) azithromycin with rifampin, PO q24h (n = 40); or (3) no antimicrobial treatment (controls; n = 41). Physical examination and thoracic ultrasonography were performed by individuals unaware of treatment group assignment. Foals that worsened were removed from the study.Results:The proportion of foals that recovered without the need to be removed from the study was significantly higher for foals treated with gamithromycin (38 of 40) or azithromycin with rifampin (39 of 40) than for controls (32 of 41). Treatment with gamithromycin or with azithromycin-rifampin resulted in a significantly faster decrease in the clinical score and abscess score compared to the controls. Adverse reactions characterized by colic (n = 18) and hind limb lameness (n = 14) were observed only in foals treated with gamithromycin.Conclusion and clinical importance:Gamithromycin was noninferior to azithromycin with rifampin for the treatment of bronchopneumonia in the study population but had a higher frequency of adverse reactions.

In the Bordetella bronchiseptica infection model development study, twenty-eight piglets were inoculated with B. bronchiseptica strain of either canine (109CFU/ml) or swine (108and 109CFU/ml) origin; swine origin strain at 109CFU/ml was chosen for the efficacy assessment study due to higher incidence and severity of gross and histopathological lesions compared with other strains. To assess efficacy of gamithromycin against B. bronchiseptica, forty piglets were experimentally inoculated on Day 0 and clinical signs were scored as per severity. Animals were then treated either with gamithromycin or saline on Day 3. The Global Clinical Scores in gamithromycin-treated group were consistently lower than the saline-treated control group from Day 4 onwards and were 0 and 40 in the gamithromycin-treated and saline-treated control groups, respectively, on Day 6. Severity and frequency of gross and histopathological observations were significantly lower in gamithromycin-treated animals compared with saline-treated controls. The efficacy of Zactran® for Swine at the label dose for the treatment of B. bronchiseptica-associated respiratory disease was demonstrated based on the faster reduction in clinical signs as early as 1 day post-gamithromycin treatment and based on the significant difference in the severity of macroscopic and microscopic lung lesions 10 days post-gamithromycin treatment.

This study assessed the plasma kinetics and skin/plasma concentration ratio of the azalide antibiotic gamithromycin (ZACTRAN(®), Merial) in sheep after a single subcutaneous administration at 6 mg/kg bodyweight. Gamithromycin concentrations in plasma samples collected at various intervals up to 21 days following treatment and metacarpal skin obtained from animals at two, five and ten days after treatment were determined by liquid chromatography-tandem mass spectrometry methods. After administration, gamithromycin was rapidly absorbed, and individual maximum plasma concentrations were observed within 6 hours post-dose. Plasma peak concentration was 573 ± 168 ng/ml. The mean area under the plasma concentration versus time curve extrapolated to infinity was 8.00 ± 1.41 µg · hr/ml, and the mean terminal half-life was 34.5 ± 5.4 hours. Gamithromycin skin concentrations were much higher than the plasma concentrations resulting in skin/plasma concentration ratios of approximately 21, 58, and 138 at two, five and ten days post-dose, respectively, demonstrating extensive distribution to skin tissue.

The aim of this study is to evaluate the safety and efficacy of gamithromycin (GAM) for the treatment of naturally occurring bacterial swine respiratory disease (SRD) administered IM. A total of 240 pigs (nine-weeks old) were selected from two sites in Heilongjiang Province of China. The pigs showed severe signs of respiratory disease. Among them, 120 pigs were randomly divided into 4 groups of low dose (3 mg/kg), middle dose (6 mg/kg), high dose (12 mg/kg) GAM IM injection and 2.5 mg/kg tulathromycin (TUL) IM injection (positive control group) for phase II clinical trial to screen effective therapeutic dose. The other 120 pigs were randomly divided into 2 groups of 6 mg/kg GAM IM injection and 2.5 mg/kg TUL IM injection (positive control group) for phase III clinical trial to further confirm the efficacy. Animals were clinically observed daily for 14 days after treatment initiation. The predominant pathogens present in pretreatment respiratory tract samples were Streptococcus suis (S. suis) and Actinobacillus pleuropneumoniae (A. pleuropneumoniae). Haemophilus parasuis (H. parasuis) and Pasteurella multocida (P. multocida) were also found in the respiratory tract. All isolates were subjected to in vitro sensitivity testing and the measured minimal inhibitory concentrations (MIC) of GAM were from 0.0625 μg/mL to 8 μg/mL. In all treatment groups, rectal temperature dropped and clinical index (mental status and respiratory symptom) significantly improved after treatment (P ≤ .05). As a result, 82.76% animals treated with the 6 mg/kg GAM injection were cured. This was significantly higher than that of 3 mg/kg GAM injection (P ≤ .05) and similar to that of 12 mg/kg GAM injection and 2.5 mg/kg TUL injection (P > .05) in phase II clinical trial. In phase III clinical trial, 80.70% of animals treated with the 6 mg/kg GAM injection were cured and the cure rate was similar to that of 2.5 mg/kg TUL injection (P > .05). In conclusion, we recommended a single dose (6 mg/kg) of GAM IM injection for the treatment of bacterial SRD.

Gamithromycin is approved for the treatment and prevention of bovine respiratory disease (BRD), which is caused mainly byMannheimia haemolytica, Pasteurella multocida, Histophilus somni, andMycoplasmaspecies. In this study, multiple dosage regimens were administered to the neutropenic mouse lung infection model in order to investigate the pharmacokinetic/pharmacodynamic (PK/PD) parameters of gamithromycin treatment ofP. multocidaand to further define the PK/PD parameter that best correlates with the efficacy of gamithromycin againstP. multocida. The PK characteristics of gamithromycin were analyzed after a single subcutaneous (s.c.) injection (1, 3, 6, and 9 mg/kg). The concentration-time profiles of unbound (f) gamithromycin in plasma samples were analyzed by non-compartmental analysis. The main PK parameters of gamithromycin for the area under the concentration-time curve from 0 to 24 h (fAUC0-24) and the peak drug concentration (f Cmax) values ranged from 0.86 to 8.42 µg·h/ml and from 0.55 to 5.69 µg/ml, respectively. The PD values were calculated based on multiple s.c. injections over 24 h (1, 3, 6, and 9 mg/kg at 6, 8, 12, and 24 h, respectively; total dosage 1-36 mg/ kg). The minimum inhibitory concentration (MIC) of gamithromycin againstP. multocidain mice serum was 0.15 μg/ml. Analysis of PK/PD indices using the inhibitory effectEmaxmodel indicated a strong correlation (R2= 0.9624) between thefAUC0-24/MIC ratio and various antibacterial effects. The area under the unbound concentration-time curve over 24 h to MIC (fAUC0-24/MIC) predicted for bacteriostatic action, 1-log10reduction, 2-log10reduction, and 3-log10reduction were 56.77, 90.18, 143.06, and 239.44 h, respectively. Thesein vivodata may facilitate gamithromycin dosage optimization againstP. multocidain veterinary medicine.

Background:Gamithromycin is a macrolide approved for the treatment of bovine and swine respiratory diseases. Our study aims to establish the clinical breakpoint and optimum dose regimen for gamithromycin against Haemophilus parasuis in piglets.Results:Gamithromycin was well absorbed and fully bioavailable (87.2-101%) after intramuscular and subcutaneous administrations. The MICs of gamithromycin for 192 clinical H. parasuis isolates ranged from 0.008 to 128 mg/L and the epidemiological cutoff (ECOFF) was calculated as 1.0 mg/L. A large potentiation effect of serum on in vitro susceptibility of gamithromycin was observed for H. parasuis, with broth/serum ratios of 8.93 for MICs and 4.46 for MBCs, respectively. The postantibiotic effects were 1.5 h (1 × MIC) and 2.4 h (4 × MIC), and the postantibiotic sub-MIC effects ranged from 2.7 to 4.3 h. Gamithromycin had rapid and concentration-dependent killing against H. parasuis, and the AUC24h/MIC ratio correlated well with ex vivo efficacy (R2= 0.97). The AUC24h/MIC targets in serum associated with bacteriostatic, bactericidal and eradication activities were 15.8, 30.3 and 41.2, respectively. The PK/PD-based population dose prediction indicated a probability of target attainment (PTA) for the current marketed dose (6 mg/kg) of 88.9% against H. parasuis. The calculated gamithromycin dose for a PTA ≥ 90% was 6.55 mg/kg. Based on Monte Carlo simulations, the PK/PD cutoff (COPD) was determined to be 0.25 mg/L.Conclusion:The determined cutoffs and PK/PD-based dose prediction will be of great importance in gamithromycin resistance surveillance and serve as an important step in the establishment of optimum dose regimen and clinical breakpoints.

A high throughput method was developed and validated for the quantitation of gamithromycin residues in eggs, milk and animal tissues (leg muscle, kidney, liver and fat) of different species and genera. This was undertaken using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The samples were extracted with acetonitrile and purified using an Oasis MCX solid phase extraction cartridge. Subsequently, a C18 column was used for chromatographic separation using acetonitrile and 0.1% formic acid as the mobile phase. LC-MS/MS in positive ESI and multiple reaction monitoring mode with gamithromycin-D4 as the internal standard was used for detection and quantification of gamithromycin. The method was successfully calibrated in the range of 1.0-200 µg/kg. The limit of detection (LOD) and limit of quantification (LOQ) for gamithromycin was 0.30-0.40 µg/kg and 0.80 - 1.0 µg/kg, respectively. The average recoveries of the analyte fortified at three levels ranged from 84.2% to 115.9%, with a relative standard deviation <10%. The proposed method has been successfully used to monitor real samples, and shown to be sensitive, rapid, and convenient. Hence, this method could be used for regulatory purposes to screen for the presence of gamithromycin residues in eggs, milk and target tissues.

In this short communication, we report the use of a second-generation macrolide antibiotic, gamithromycin (Gam), as a novel chiral selector for enantioseparation in capillary electrophoresis (CE). A preliminary analysis of the experiment results shows that Gam is especially suitable for the separation of chiral primary amines. Factors influencing enantioseparations were systematically investigated including the composition of the background electrolyte (BGE), concentration of Gam, the type and proportion of organic solvents, applied voltage, etc. In particular, N-Methylformamide (NMF) was successfully used as a non-aqueous solvent for Gam, and shown to be extremely effective for the separation of primaquine (PMQ) and 1-aminoindan (AMI) when used alone or mixed with other commonly used non-aqueous solvents (e.g. methanol). To our knowledge this was also the first application of NMF as a non-aqueous solvent for antibiotic chiral selectors in CE. The best separations were obtained with 100 mM Tris, 125 mM H3BO3and 80 mM Gam in methanol/NMF (25:75) solvent for PMQ and AMI, or 80-100 mM Gam in methanol for the other model analytes. Among the analytes, the resolution (Rs) of amlodipine (AML) reached up to 15.65, which is to our knowledge the highest value ever reported in CE studies for this compound (except for using molecularly imprinted polymers technique).

The pharmacokinetics of gamithromycin were evaluated in 26 male castrated and female crossbred swine administered gamithromycin 15% w/v (Zactran®, Boehringer Ingelheim) intravenously at 6 mg/kg bodyweight or intramuscularly at 3, 6 or 12 mg/kg bodyweight. Blood samples were collected up to Day 10 to establish the plasma profile of gamithromycin, bioavailability and dose proportionality. When administered by intramuscular injection at 6 mg/kg BWT, pharmacokinetic parameters were as follows: area under the curve until last quantifiable plasma concentration, 5.13 ± 0.957 µg*hours/ml; maximum plasma concentration, 960 ± 153 ng/ml at 5 to 15 min; terminal half-life of 94.1 ± 20.4 hr. Absolute bioavailability was 92.2%. Increase in systemic exposure was proportional to the gamithromycin dose level over the range 3-12 mg/kg BWT. No gender-related statistically significant difference in exposure was observed. For clinical evaluation of Zactran® against swine respiratory disease, 305 pigs from six commercial farms in three countries in Europe with signs associated with Actinobacillus pleuropneumoniae and/or Haemophilus parasuis and/or Pasteurella multocida and/or Bordetella bronchiseptica were used. At each site, animals were treated once in a 1:1 ratio with a single intramuscular dose of Zactran® (6 mg gamithromycin/kg bodyweight) or Zuprevo® (4% w/v tildipirosin at 4 mg/kg bodyweight; MSD Animal Health) at the recommended dose respectively. Animals were observed and scored daily for 10 consecutive days for signs of swine respiratory disease (depression, respiration and rectal temperature), and animals presenting signs of clinical swine respiratory disease (Depression Score 3 and/or Respiratory Score 3 associated with Rectal Temperature > 40.0°C) were removed from the study and considered as treatment failure. Animals which remained in the study were individually assessed for 'treatment success' or 'treatment failure' (Depression Score ≥ 1 and Rectal Temperature > 40.0°C or Respiratory Score ≥ 1 and Rectal Temperature > 40.0°C). Using a non-inferiority hypothesis test (non-inferiority margin = 0.10), the proportion of treatment successes in the Zactran® group (97%) was equivalent to or better than that in the Zuprevo® group (93%).

In the context of a development program to obtain the market authorization of injectable gamithromycin 15% w/v solution (Zactran®, Boehringer Ingelheim) for use in sheep against footrot, the pharmacokinetic profile of gamithromycin was established and the safety and efficacy of the treatment were confirmed in a multicenter field study in Europe. The basic pharmacokinetic parameters established in healthy young Merino sheep administered gamithromycin at 6 mg/kg body weight based on the analysis of plasma samples which were collected in intervals up to 12 days after subcutaneous injection were: area under the curve until last quantifiable concentration, 8.88 ± 2.33 μg*h/mL; maximum plasma concentration, 448 ± 180 ng/mL; terminal half-life, 42.5 ± 5.25 h. The safety and clinical efficacy against footrot of gamithromycin 15% w/v solution were evaluated in comparison to tilmicosin 30% w/v solution (Micotil®, Elanco) treatment in 364 sheep of various breeds, sex and age from commercial farms in the United Kingdom (2 sites), Germany (3 sites) and France (1 site). Animals were enrolled based on lesions characteristic of footrot and lameness associated with the presence of footrot-related bacterial pathogens and were randomly allocated and treated in a 1:1 ratio with a single subcutaneous dose of gamithromycin or tilmicosin at label dosage (6 or 10 mg/kg body weight, respectively). Lameness and footrot lesions were evaluated at five and 21 days after treatment; the injection site in all animals was examined the day after treatment and followed up daily in the animals with injection site reaction until complete injection site reaction resolution. Samples of 310 and 120 animals tested positive for Dichelobacter nodosus and Fusobacterium necrophorum, respectively, at inclusion, and data of 359 animals were included into the combined analyses (5 animals excluded for unintentional overdosing [1], lack of follow-up [1], concurrent antibiotic medication for non-footrot conditions [3]). Lameness scores at 21 days after treatment demonstrated a significantly (p = 0.0396) better success for the gamithromycin treatment compared to the tilmicosin treatment (97.8% vs. 93.3%). Post-dosing footrot lesion scores followed similar trends of rapid and marked decrease (improvement) for both treatments with similar (p = 0.127) treatment success for the gamithromycin and tilmicosin treatments (97.8% and 96.0%, respectively). Both treatments were safe; injection site reactions noted in 19 gamithromycin- and 25 tilmicosin-treated animals resolved within five days or six days of treatment, respectively. Gamithromycin 15% w/v solution administered once to sheep by subcutaneous injection at 6 mg/kg body weight demonstrated a pharmacokinetic profile similar to that reported previously in sheep and cattle and was confirmed to be a safe and efficacious treatment for naturally occurring ovine footrot in a multicenter clinical field study conducted in Europe.