Coccidia; nticoccidial

Nicarbazin (NCZ) is an anticoccidial drug routinely used in the poultry industry that can negatively affect reproduction by reducing egg production, egg weight, and egg hatchability. The molecular mechanisms by which NCZ affects reproduction are unknown. Lipoprotein lipase, vitellogenin, transglutaminase, and calcium are all involved in egg formation and embryogenesis. Therefore, in vitro assays were used to evaluate 4 potential mechanisms of action of NCZ on egg formation and embryogenesis. First, a lipoprotein lipase assay was conducted to determine if NCZ increases lipoprotein lipase activity. Second, vitellogenin phosphorylation was evaluated to determine if NCZ acts as a vitellogenin phosphatase. Third, transglutaminase activity was measured to determine if NCZ inhibits transglutaminase activity. Finally, bull sperm was used as a model to determine if specific channel-mediated calcium uptake can be blocked by NCZ. Nicarbazin increased the activity of lipoprotein lipase in vitro at 3.9 and 7.8 microg of NCZ/mL. Nicarbazin increased intracellular calcium levels in bull sperm, suggesting it also acts as a calcium ionophore. The portion of the NCZ molecule responsible for the increase in intracellular calcium is 2-hydroxy-4,6-dimethylpyrimidine. Nicarbazin affected vitellogenin phosphorylation but only at a concentration many times higher than expected plasma values. Nicarbazin also inhibited transglutaminase activity in vitro. Whereas the 4,4'-dinitrocarbanilide portion of the NCZ molecule inhibited transglutaminase activity, the 2-hydroxy-4,6-dimethylpyrimidine portion increased transglutaminase activity. All of these assays were conducted in vitro; therefore these results should be viewed as preliminary findings to aid in directing further research on the effect of NCZ on reproduction in vivo. Because NCZ increases lipoprotein lipase activity and acts as a calcium ionophore, future experiments should investigate these effects in particular.https://pubmed.ncbi.nlm.nih.gov/16830870/

Nicarbazine has no negative effects on immune development or resistance to Eimeria curvularis oocyst reinfection [1].

Absorption, Distribution and Excretion
Rats were administered nicarbazine orally at doses of 1, 5, or 10 mg/kg body weight. One animal from each dose group was sacrificed 6 or 18 hours after administration, and the blood concentrations of phenylurea and pyrimidinone components were measured. Low concentrations of phenylurea were detected at 6 hours, but not at 18 hours. The concentration of pyrimidinone was significantly higher than that of phenylurea and remained elevated between 6 and 18 hours. Similar qualitative results were obtained when rats were administered nicarbazine orally at 0.1, 1, or 5 mg/kg body weight daily for 8 consecutive days, and sacrificed 4 or 24 hours after the last administration. The blood concentration of pyrimidinone was dose-dependent, while the blood concentration of phenylurea showed no significant change. In the latter experiment, the concentrations of all nicarbazine components in urine collected within 5 hours of the last administration were dose-dependent, but the concentration of pyrimidinone was an order of magnitude higher than that of phenylurea. Of the 190 eggs analyzed, nicarbazine was detected in 39 eggs in the form of 4,4'-dinitrocarbaniline (DNC). The concentrations of both DNC and 4,6-dimethyl-2-hydroxypyrimidine (DHP) in the eggs were directly proportional to feed levels. The highest concentration of nicarbazine in the feed was 12.1 mg/kg (8.6 mg/kg of DNC and 3.5 mg/kg of DHP), resulting in average highest concentrations of DNC and DHP in the eggs of 631 μg/kg and 51.8 μg/kg, respectively. After discontinuation of the experimental feed, DNC was undetectable in the eggs after 12 days, and DHP was undetectable after 3 days. When the nicarbazine concentration in the feed exceeded approximately 2 mg/kg, the residual concentration of dinitrobenzoic acid (DNC) in the eggs exceeded the UK-set differential action limit (DAL, 100 μg/kg). DNC is almost entirely found in the yolk, while dihydropyridine (DHP) is distributed in the egg white and yolk in a ratio of approximately 3:1.

Adverse Reactions
Occupational Hepatotoxicity - Secondary Hepatotoxicity: Potential toxic effects in the occupational environment based on cases of human ingestion or animal poisoning. Hazardous Chemicals and Occupational Disease Information, Haz-Map

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Antidotes and Emergency Treatment
Basic Treatment: Maintain an open airway (use an oropharyngeal or nasopharyngeal airway if necessary). Suction if necessary. Observe for signs of respiratory failure and provide assisted ventilation if necessary. Administer oxygen via a non-invasive breathing mask at a flow rate of 10 to 15 liters per minute. Monitor for pulmonary edema and treat if necessary… Monitor for shock and treat if necessary… Prevent seizures and treat if necessary… If eyes are contaminated, flush with water immediately. During transport, continuously flush each eye with 0.9% saline… Do not use emetics. In case of ingestion, rinse mouth and dilute with 5 ml/kg to 200 ml of water, provided the patient is able to swallow, has a strong gag reflex, and does not drool… Skin burns should be disinfected and covered with a dry, sterile dressing… /Class A and B Poisons/
Advanced Treatment: For patients with altered mental status, severe pulmonary edema, or severe respiratory distress, consider oropharyngeal or nasopharyngeal endotracheal intubation to control the airway. Positive pressure ventilation with a bag-valve-mask may be effective. Consider medical treatment for pulmonary edema… Consider the use of a beta-agonist (such as salbutamol) for severe bronchospasm… Monitor heart rhythm and treat arrhythmias if necessary… Initiate intravenous infusion of 5% glucose solution (SRP: “Keep it patent,” minimum flow rate). If signs of hypovolemia appear, use 0.9% normal saline or lactated Ringer's solution. Use fluids with caution in cases of hypotension with signs of hypovolemia. Watch for signs of fluid overdose… Use diazepam or lorazepam for seizures… Use promecaine hydrochloride to assist eye irrigation… Toxins A and B /

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Non-human toxicity values
Oral LD50 in mice: approx. 4,000 mg/kg / 2-Hydroxy-4,6-dimethylpyrimidine (HDP) /
Oral LD50 in rats: > 10,000 mg/kg body weight
Oral LD50 in mice: > 18,000 mg/kg body weight / 4,4'-dinitrocabanelli (DNC) /
Oral LD50 in mice: > 25,000 mg/kg body weight

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Non-human toxicity values
Oral LD50 in mice: approx. 4,000 mg/kg / 2-Hydroxy-4,6-dimethylpyrimidine (HDP) /
Oral LD50 in rats: > 10,000 mg/kg body weight
Oral LD50 in mice: > 18,000 mg/kg body weight / 4,4'-dinitrocabanelli (DNC) /
Oral LD50 in mice: > 18,000 mg/kg body weight > 25,000 mg/kg body weight
Rat skin LD50 > 5,000 mg/kg body weight

[1]. Ott, W. H., et al. Biological Studies on Nicarbazin, a New Anticoccidial Agent. Poultry Science. 1956. 35(6), 1355–1367.
[2]. CUCKLER AC, et al. The effect of nicarbazin on the development of immunity to avian coccidia. J Parasitol. 1956 Dec;42(6):593-607.

Nicarbazin is an organic molecular entity. It is an equimolar complex of 4,4'-dinitrocarbaniline and 2-hydroxy-4,6-dimethylpyrimidine, and is an anticoccidial agent used in poultry. See also: 4,4'-dinitrocarbaniline (with active moiety); nalazine; nicarbazin (ingredient); bacitracin zinc; nicarbazin (ingredient)... See more...

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Mechanism of Action
Nicarbazin interferes with the formation of the vitelline membrane, which separates the yolk from the albumen. Its exact mechanism of action is unclear, but it is believed that nicarbazin interferes with cholesterol metabolism during vitelline membrane formation. Eggs from nicarbazin-treated poultry have a mottled appearance, reflecting the porosity of the vitelline membrane. The effect on hatchability depends on time and dosage, and this effect is reversible.

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Therapeutic Uses
/VET:/ Nicarbazin has been used in chick diets for decades to prevent fecal and intestinal coccidiosis in broilers. It can be used in combination with ionotropic coccidioidomycin inhibitors. /VET:/ This anticoccidial drug /nicarbazin/ is used in broilers.

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Drug Warnings
/VET:/ should not be fed to laying hens as it causes egg discoloration and reduced hatchability (although this effect is reversible after discontinuation of nicarbazin). It may also cause reduced heat tolerance in poultry exposed to high temperature and humidity;

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Therapeutic Use
/VET:/ nicarbazin has been used in chick diets for decades to prevent fecal and intestinal coccidiosis in broilers. It can be used in combination with ionotropic anticoccidial drugs. /VET:/ This anticoccidial drug /nicarbazin/ is used in broilers.

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Drug Warnings
/VET:/ should not be fed to laying hens as it causes egg discoloration and reduced hatchability (although this effect is reversible after discontinuation of nicarbazin). It may also cause reduced heat tolerance in poultry exposed to high temperature and humidity;

C19H18N6O6

426.39

426.128

C, 53.52; H, 4.26; N, 19.71; O, 22.51

330-95-0

Nicarbazin-d8

9507

Crystals
Light yellow, fine powder

1.4663 (rough estimate)

414.8ºC at 760 mmHg

265-275ºC

204.7ºC

1.6000 (estimate)

5.138

3

6

2

31

582

0

C1=C(C=CC(=C1)[N+](=O)[O-])NC(=O)NC2=CC=C(C=C2)[N+](=O)[O-].CC1=CC(=NC(=N1)O)C

UKHWDRMMMYWSFL-UHFFFAOYSA-N

InChI=1S/C13H10N4O5.C6H8N2O/c18-13(14-9-1-5-11(6-2-9)16(19)20)15-10-3-7-12(8-4-10)17(21)22;1-4-3-5(2)8-6(9)7-4/h1-8H,(H2,14,15,18);3H,1-2H3,(H,7,8,9)

1,3-bis(4-nitrophenyl)urea;4,6-dimethyl-1H-pyrimidin-2-one

Nicoxin; AI3-60130; Nicarbasin; Nirazin; Nicarbazine; HSDB 7466; Nicarbazin; MK 75; Nicarb; Nicrazin; Nicrazine

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 : ~8.93 mg/mL (~20.94 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.)