Bacterial protein synthesis; Microbial Metabolite; Bacterial HSV-1; Endogenous Metabolite

Oxytetracycline calcium is a key component of the bacterial aromatic polyketide family, a group of natural compounds with a variety of topologies. Oxytetracycline calcium is generated by type II polyketide synthase, which produces a poly-β-ketone backbone through consecutive decarboxylation condensation of malonyl-CoA extension units. The backbone is subsequently passed through cyclases, oxygenases, transferases, and other tailoring enzymes. Make adjustments [2].

The effects of calcium delivery of therapeutic dosages of oxytetracycline (82.8 mg/kg body weight to 1% body weight/day) for 10 days were species specific. Oxytetracycline calcium increases the relative liver weight of Morone chrysops For oxytetracycline calcium, the limits are 100 μg/kg in muscle and milk, 200 μg/kg in eggs, 300 μg/kg in liver and 600 μg/kg in kidneys. Oxytetracycline calcium is supplied to fish as medicated feed at dosages ranging from 35 to 75 mg ai kg-1 biomass on day 1 and continued for 7-14 days [1].

Oxytetracycline (OTC) is a broad-spectrum antibiotic that acts by inhibiting protein synthesis in bacteria. It is an important member of the bacterial aromatic polyketide family, which is a structurally diverse class of natural products. OTC is synthesized by a type II polyketide synthase that generates the poly-beta-ketone backbone through successive decarboxylative condensation of malonyl-CoA extender units, followed by modifications by cyclases, oxygenases, transferases, and additional tailoring enzymes. Genetic and biochemical studies have illuminated most of the steps involved in the biosynthesis of OTC, which is detailed here as a representative case study in type II polyketide biosynthesis[2].

1. Viral Plaque Reduction Assay - Cell Line: Vero cells infected with HSV-1 (MOI = 0.1). - Protocol: 1. Treat cells with oxytetracycline (0.1–1 mg/mL) 2 hours post-infection [3]
2. Incubate for 48 hours and fix with formaldehyde [3]
3. Stain with crystal violet and count plaques [3]
- Results: Oxytetracycline (1 mg/mL) reduced plaque count by 70% compared to control [3]

Oxytetracycline (OTC) is employed in fish farms to contest or prevent bacterial infections. We simulated an OTC treatment at therapeutic level (75 mg kg(-1)) and at higher doses (150, 300 mg kg(-1)) for 10 days. A withdrawal period of 10 days was considered for treated carp, carrying out the same chemical and biochemical analyses (total glutathione, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase and malondialdehyde). The aim was to obtain data related to the carryover in muscle and on variations in the antioxidant indicators in liver and kidney. The OTC residual levels in muscle showed a dose-response relationship. After 10 days of treatment at the recommended dose (75 mg kg(-1)), the mean value in muscle was 295 μg kg(-1). After 10 withdrawal days, residues in all treated groups were not entirely eliminated by fish. Residues of recommended 75 mg kg(-1) OTC dose were lower than the maximum permitted by EEC regulation: 100 μg kg(-1). Disturbance in the antioxidant systems in liver and kidney was recorded in (150, 300 mg kg(-1)) carp, as well as during the withdrawal period. A lowered superoxide dismutase activity and higher levels of catalase, glutathione peroxidase, glutathione reductase and glutathione were evaluated in liver, while in kidney only higher malondialdehyde and glutathione S-transferase concentrations were recorded for 300 mg kg(-1) dose. The therapeutic OTC dose exerted lower effects, and only in liver, enhancement of GPx and GR activities was recorded. After the withdrawal period, altered antioxidant responses in tissues were restored for all three OTC doses.[1]

Absorption, Distribution and Excretion
Oxytetracycline is readily absorbed after oral administration. The serum half-life in horses is 15.7 hours after intravenous injection and 10.5 hours after intramuscular injection. Dose-dependent kinetics may be a contributing factor. The absorption rate of the fasting oral dose is 60% to 80% for oxytetracycline. Peak plasma concentrations of oxytetracycline are reached within 2 to 4 hours after a single oral dose. Its half-life is 6 to 12 hours, and it is typically administered 2 to 4 times daily. A dose of 250 mg every 6 hours results in a peak plasma concentration of 2 to 2.5 μg/mL. Doses exceeding 1 g every 6 hours do not significantly increase plasma concentrations. Approximately 10% to 35% of oxytetracycline is excreted in the active form in the urine, detectable within 30 minutes, and reaches peak concentration approximately 5 hours after administration. Oxytetracycline binds to plasma proteins at a rate of approximately 20% to 25%. Absorption in the lower digestive tract is far from complete; bile concentrations are 5 to 10 times higher than plasma concentrations. /Tetracyclines/
For more complete data on the absorption, distribution, and excretion of oxytetracyclines (20 in total), please visit the HSDB records page.

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Biological half-life
Biological half-life…may be 3–4 days in patients with anuria.
In adults with normal renal function, the serum half-life of oxytetracycline is 6 to 10 hours; it has been reported to be 47 to 66 hours in patients with severe renal impairment. In patients with normal renal function, approximately 60% to 70% of the active drug is excreted in the urine within 72 hours after a single oral dose of oxytetracycline.
A two-way crossover trial was conducted in 6–8 month old crossbred male calves to determine the bioavailability, pharmacokinetics, and dosing regimen of long-acting oxytetracycline formulations (OTC-LA). The half-lives of oxytetracycline were 7.8 hours and 24 hours after intravenous and intramuscular administration, respectively. …
The pharmacokinetic properties of oxytetracycline were investigated after a single intramuscular injection of a long-acting oxytetracycline formulation (20 mg/kg body weight) into the semimembranosus of healthy dogs and experimentally infected dogs with Ehrlichia canis. …The mean apparent elimination half-life (t_1/2β) was significantly prolonged after infection. The absorption half-life (t(1/2) ab) was significantly shortened after infection.
The serum half-life in horses… was 15.7 hours after intravenous administration and 10.5 hours after intramuscular administration. …Dose-dependent kinetics may be one factor.

Effects During Pregnancy and Lactation
◉ Overview of Drug Use During Lactation
Some reviews suggest that tetracyclines are contraindicated during lactation because they may cause staining of infant tooth enamel or deposition in bone. However, a careful review of existing literature indicates that short-term use of oxytetracycline during lactation is unlikely to cause harm because the drug concentration in breast milk is low, and the infant's absorption of the drug is inhibited by calcium in breast milk. Short-term use of oxytetracycline in lactating women is acceptable. As a theoretical precaution, prolonged or repeated use during lactation should be avoided. Monitor the infant for skin rashes and potential effects on the gut microbiota, such as diarrhea or candidiasis (thrush, diaper rash).
◉ Effects on Breastfed Infants
No adverse reactions were observed in breastfed infants receiving oral oxytetracycline 1.5 or 2 g/day for 3 consecutive days. The infant's age and degree of breastfeeding were not considered.
◉ Effects on Lactation and Breast Milk
No relevant published information was found as of the revision date.

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Interactions
Concomitant administration of ferrous sulfate reduces the absorption of oxytetracycline in the human body, leading to a significant decrease in serum oxytetracycline concentration. Concomitant consumption of milk reduces oxytetracycline absorption by approximately 50%...
Bromhexine treatment leads to an increase in oxytetracycline concentration in nasal mucus...This is attributed to mucus evaporation due to decreased viscosity after bromhexine treatment, rather than increased membrane permeability.
...Oxytetracycline may cause unpredictable fluctuations in blood glucose levels by prolonging the insulin half-life......Furthermore...when oxytetracycline and tolbutamide...are used concurrently.
In the clinical treatment of pneumococcal meningitis, significant antagonistic effects have been observed between penicillin and tetracycline.../tetracycline/
For more complete data on interactions of oxytetracyclines (10 in total), please visit the HSDB record page.

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Non-human Toxicity Values
Oral LD50 in Swiss mice: 7200 mg/kg /hydroxytetracycline hydrochloride/

[1]. Transferability of oxytetracycline (OTC) from feed to carp muscle and evaluation of the antibiotic effects on antioxidant systems in liver and kidney. Fish Physiol Biochem, 2014 Aug;40(4):1055-68.

[2]. Oxytetracycline biosynthesis. J Biol Chem. 2010 Sep 3;285(36):27509-15.

[3]. A New Treatment Method for Herpes Simplex Virus Type 1-related Skin Lesions. Scientific & Academic. 2019; 8(1): 6-8.

Depending on state or federal labeling requirements, oxytetracycline (oral) may cause developmental toxicity. Oxytetracycline is a tetracycline antibiotic used to treat infections caused by a variety of Gram-positive and Gram-negative bacteria, including Mycoplasma pneumoniae, Pasteurella multocida, Escherichia coli, Haemophilus influenzae (respiratory infections), and Diplococcus pneumoniae. It has multiple actions, including antibacterial activity, protein synthesis inhibition, antimicrobial activity, anti-inflammatory activity, and bacterial metabolite activity. It is a zwitterionic tautomer of oxytetracycline. An anhydrous oxytetracycline is a tetracycline antibacterial agent. Oxytetracycline has been reported to be found in Streptomyces anthocyanicus, Streptomyces varsoviensis, and other microorganisms with relevant data.
Oxytetracycline, a tetracycline analog isolated from the actinomycete Streptomyces rimosus, is widely used for a variety of clinical diseases.
Drug Indications

Oxytetracycline is indicated for the treatment of infections caused by a variety of Gram-positive and Gram-negative bacteria, including Mycoplasma pneumoniae, Pasteurella multocida, Escherichia coli, Haemophilus influenzae (respiratory tract infection), and Streptococcus pneumoniae.
Mechanism of Action

Oxytetracycline inhibits cell growth by inhibiting translation. It binds to the 30S ribosomal subunit, preventing aminoacyl-tRNA from binding to the A site of the ribosome. This binding is reversible. Oxytetracycline is lipophilic and can easily cross the cell membrane or passively diffuse through porin channels on the bacterial membrane.
Tetracycline drugs inhibit bacterial protein synthesis by binding to the bacterial 30S ribosome, preventing aminoacyl-tRNA from binding to the receptor (A) site on the mRNA-ribosome complex. They passively diffuse into Gram-negative bacteria through hydrophilic channels formed by porins in the outer cell membrane, or actively transport via an energy-dependent system that pumps all tetracyclines into the cytoplasmic membrane. Although the mechanisms by which these drugs penetrate Gram-positive bacteria are poorly understood, this process also requires energy. At high concentrations, these compounds inhibit protein synthesis in mammalian cells. However, because mammalian cells lack the active transport systems present in bacteria and have lower sensitivity to ribosome targets, tetracyclines exhibit selective activity against bacteria. Tetracycline antibiotics… may exert neuromuscular blocking effects through chelation of Ca²⁺. /Tetracyclines/

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Therapeutic Uses
MeSH Title: Antibacterial Agents
Antibiotics, Tetracyclines
...broad-spectrum antibacterial activity against Gram-positive and Gram-negative bacteria...against some microorganisms naturally insensitive to many chemotherapeutic agents, such as Rickettsia, Mycoplasma, Chlamydia, lymphogranuloma venereum, psittacosis, inclusion conjunctivitis and trachoma pathogens, and amoebae. /Tetracyclines/
Tetracyclines are active against a wide variety of aerobic and anaerobic Gram-positive and Gram-negative bacteria. They are also effective against some microorganisms resistant to cell wall-active antibacterial agents, such as Rickettsia, Coxsella burgdorferi, Mycoplasma pneumoniae, Chlamydia spp., Legionella spp., Ureaplasma, some atypical mycobacteria, and Plasmodium spp. They are ineffective against fungi. Tetracyclines
For more complete data on the therapeutic uses of oxytetracyclines (28 in total), please visit the HSDB record page.

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Drug Warnings
While individual differences have not shown significant variations except in a few extreme cases, some oxytetracycline preparations have been shown to be ineffective against generic drugs.
Food, milk, non-systemic antacids, and iron supplements can interfere with oral absorption.
…Ineffective against any true virus, yeast, or fungus. Tetracyclines
Due to the high risk of sensitization, topical application is best avoided, except for ophthalmic application…Intrathecal injection is absolutely prohibited. /Tetracyclines/
For more complete data on drug warnings for oxytetracyclines (38 in total), please visit the HSDB record page.

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Pharmacodynamics
Oxytetracycline is known as a broad-spectrum antibiotic due to its activity against a variety of infections. It was the second tetracycline to be discovered. Like other tetracyclines, oxytetracycline is used to treat a variety of common and rare infections. Its superior absorption properties make it a preferred treatment for moderate to severe acne, but if the condition does not improve after 3 months, alternative medications should be sought.

C22H22CAN2O9

500.51

498.095

7179-50-2

Oxytetracycline;79-57-2;Oxytetracycline hydrochloride;2058-46-0;Oxytetracycline dihydrate;6153-64-6

54675779

Light-yellow crystals or needles from aqueous MeOH
Pale yellow to tan, crystalline powder

0.06

7

10

2

33

1000

6

[Ca+2].NC(C1C(=O)[C@@H](N(C)C)C2[C@H]([C@@H]3[C@@](O)(C)C4=CC=CC([O-])=C4C([O-])=C3C(=O)[C@]2(O)C=1O)O)=O

OWFJMIVZYSDULZ-PXOLEDIWSA-N

InChI=1S/C22H24N2O9/c1-21(32)7-5-4-6-8(25)9(7)15(26)10-12(21)17(28)13-14(24(2)3)16(27)11(20(23)31)19(30)22(13,33)18(10)29/h4-6,12-14,17,25-26,28,30,32-33H,1-3H3,(H2,23,31)/t12-,13-,14+,17+,21-,22+/m1/s1

(4S,4aR,5S,5aR,6S,12aR)-4-(dimethylamino)-1,5,6,10,11,12a-hexahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide

Oxytetracycline calcium; Calcium oxytetracycline; Terrabon; Dioxytetracycline calcium; ...; 7179-50-2;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)