β-lactam; cell wall synthesis

Isolated microspore culture is a promising option to rapidly fix the product of meiotic recombination of F1 hybrids, in the process of varietal development. Clean culture and high embryogenesis rate are essential to commercial triticale and wheat microspore cultures. So, this study investigated (1) contaminants from isolated microspores cultures, (2) two antibiotics to control bacterial growth, and (3) the contribution of antibiotics to increased microspore-derived embryo-like structures (ELS), green and albino plants. Five species of bacteria were identified in contaminated cultures (Erwinia aphidicola, Pantoea agglomerans, Pseudomonas sp., Staphylococcus epidermis and Staphylococcus warneri) using fatty acid analysis and 16S ribosomal RNA sequences analysis, and yeast. Antibacterial susceptibility test using Cefotaxime and Vancomycin resulted in strong inhibition of 24 bacterial isolates, using Cefotaxime at 100 mg/l, but not Pseudomonas sp. Other antibiotic treatments inhibited bacterial growth at least partially. Microspore induction medium supplemented with the same antibiotics treatments resulted in successful microspore embryogenesis and green plant production. Antibiotic treatments were first tested in triticale and then validated in wheat cultivars AC Carberry and AC Andrew. Induction medium supplemented with Cefotaxime at 50 and 100 mg/l substantially increased the formation of ELS and green plants in triticale and wheat, respectively. Incidentally, it also affected the occurrence of albinism in all genotypes. Our results demonstrated dual purpose of Cefotaxime for isolated microspore culture, most importantly it increases cell growth and success of microspore cultures in triticale and wheat genotypes, but would also prevent accidental loss of cultures with most common bacterial contaminants[3].

In rats infected with neutropenic Pseudomonas, carbenicillin (100–400 mg/kg; intramuscularly every 8 hours for 72 hours) dramatically lowers mortality [1].

The isolates were grown for 48–72 h at 27 °C on LB medium in individual Petri dishes that were supplemented with different antibiotics (Table 1) to evaluate colony sensitivity towards antibiotics. The following antibiotics treatments were applied into the antibiotic assay on isolates and in isolated microspores cultures of triticale and wheat genotypes: T1: Control (no antibiotic); T2: Vancomycin at 100 mg/l; T3: Vancomycin 500 mg/l; T4: Cefotaxime at 50 mg/l; T5: Cefotaxime 100 mg/l; T6: Vancomycin 100 mg/l and Cefotaxime 50 mg/l; and T7: Vancomycin 500 mg/l and Cefotaxime 100 mg/l. The isolates’ growth was noted as no inhibition (+++), weak inhibition (++), strong inhibition (+) and no growth (−) relative to control, where no antibiotic was applied[3].

Rats made neutropenic with cyclophosphamide were infected intraperitoneally with Pseudomonas aeruginosa. The challenge organism was killed synergistically in vitro by the combination of gentamicin and carbenicillin. Untreated neutropenic rats infected with 3 x 10(6)Pseudomonas died between days 2 and 7, and the overall mortality was 70%. Groups of infected neutropenic rats were treated intramuscularly with 1.5 or 6 mg of gentamicin per kg per dose, 100 or 400 mg of carbenicillin per kg per dose, or 1.5 mg of gentamicin and 100 mg of carbenicillin per kg per dose. Treatment was begun at 2 h postinfection and was continued every 8 h for about 72 h. Cultures of blood and peritoneal washings were performed in control and treated rats at 1, 4, 24, 48, and 72 h postinfection. Gentamicin at either dose level was ineffective in preventing death, but mortality was significantly reduced by high-dose carbenicillin and low-dose combination therapy. In addition, the latter regimens sterilized the peritoneal fluid and blood. Carbenicillin and gentamicin showed in vivo synergy in the treatment of neutropenic Pseudomonas-infected rats[1].

Absorption, Distribution and Excretion
Carbenicillin is rapidly absorbed through the small intestine after oral administration. Oral bioavailability is 30% to 40%. Carbenicillin is not absorbed through the gastrointestinal tract and therefore must be administered parenterally. …After intramuscular injection of 1 gram, peak plasma concentrations reach 15-20 μg/mL within 0.5-2 hours…After intravenous administration, peak plasma concentrations are approximately four times higher than after intramuscular injection…After intravenous infusion…after 1 gram/hour, the average plasma concentration is approximately 150 μg/mL. Approximately 50% of the antibiotic in plasma is bound to proteins. …Carbenicillin is primarily excreted via the renal tubules. Approximately 75-80% is recovered in the urine in its active form within 9 hours. Carbenicillin is unstable in acidic environments and is intolerant to penicillinase; therefore, due to poor oral absorption, parenteral administration is usually used. The parenteral dose is 50-200 mg/kg, administered in divided doses every 4-6 hours.
Although carbenicillin is cleared from bovine mammary glands within 72 hours after treatment, it is ineffective in treating mastitis caused by Pseudomonas aeruginosa; resistant bacteria can be recovered 24–48 hours after infusion.
For more complete data on the absorption, distribution, and excretion of carbenicillin (11 in total), please visit the HSDB record page.

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Metabolism/Metabolites
Very few.

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Biological half-life
1 hour
...T 1/2...Approximately 1 hour in individuals with normal renal function...Extended to approximately 2 hours in individuals with hepatic impairment.

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Limited information suggests that carbenicillin concentrations in breast milk are low and are not expected to have adverse effects on breastfed infants. There are reports that penicillin drugs occasionally disrupt the infant's gut microbiota, leading to diarrhea or thrush, but these effects have not been fully assessed. Carbenicillin indane disodium can be used in breastfeeding women. ◉ Effects on Breastfed Infants
No published information found as of the revision date. ◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.

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Protein Binding Rate
30% to 60%Interactions
Broad-spectrum penicillins are physically and/or chemically incompatible with aminoglycosides and can inactivate aminoglycosides in vitro. The in vitro inactivation of aminoglycoside antibiotics by penicillin depends on the specific drug and appears to be directly proportional to penicillin concentration, exposure time, and temperature… Most in vitro studies have shown that carbenicillin inactivates aminoglycoside antibiotics faster than maloxicillin, piperacillin, or ticarcillin. Broad-spectrum penicillins can also inactivate aminoglycoside antibiotics in vivo. In patients with impaired renal function, concomitant use of carbenicillin and gentamicin reduces serum aminoglycoside antibiotic concentrations and serum half-life compared to aminoglycoside antibiotics alone. Some in vitro studies suggest that the antibacterial activity of broad-spectrum penicillins may have additive or partially synergistic effects with other β-lactam antibiotics… /Broad-spectrum penicillin/ In vitro studies have shown that carbenicillin, in combination with the β-lactamase inhibitor clavulanate, has a synergistic bactericidal effect against a variety of β-lactamase-producing bacterial strains. For more complete data on interactions of carbenicillin (7 types), please visit the HSDB record page.

[1]. Scott RE, et, al. Synergistic activity of carbenicillin and gentamicin in experimental Pseudomonas bacteremia in neutropenic rats. Antimicrob Agents Chemother. 1976 Oct;10(4):646-51.
[2]. Neu HC, et, al. Carbenicillin and ticarcillin. Med Clin North Am. 1982 Jan;66(1):61-77.
[3]. Cefotaxime prevents microbial contamination and improves microspore embryogenesis in wheat and triticale. Plant Cell Rep . 2013 Oct;32(10):1637-46.

Carbenicillin is a penicillin antibiotic with a side chain of 6β-2-carboxy-2-phenylacetamido. It is an antibacterial drug. It is both a penicillin and a penicillin allergen. It is the conjugate acid of carbenicillin (2-). It is a broad-spectrum semi-synthetic penicillin derivative that can be administered parenterally. It is susceptible to gastric juice and penicillinase and may impair platelet function. Carbenicillin belongs to the penicillin class of antibacterial drugs. There have been reports of carbenicillin being detected in tomato (Solanum lycopersicum), and relevant data exist. Carbenicillin is a broad-spectrum semi-synthetic penicillin antibiotic with bactericidal activity and resistance to β-lactamases. Carbenicillin works by opening the c-terminal domain of penicillin-sensitive transpeptidase. This inactivation prevents the cross-linking of peptidoglycan chains, thereby inhibiting the third (and final) stage of bacterial cell wall synthesis. This leads to incomplete bacterial cell wall synthesis, ultimately resulting in cell lysis.
A broad-spectrum semi-synthetic penicillin derivative, administered parenterally. It is susceptible to gastric juice and penicillinase and may impair platelet function.
Indications

For the treatment of acute and chronic upper and lower urinary tract infections caused by susceptible bacterial strains, as well as asymptomatic bacteriuria.
Mechanism of Action

Free carbenicillin is the main pharmacologically active component of this salt. Carbenicillin exerts its antibacterial activity by interfering with the final cell wall synthesis of susceptible bacteria. Penicillin opens the C-terminal domain of a penicillin-sensitive transpeptidase via a lactam ring. Inactivation of this enzyme prevents the formation of cross-links between two linear peptidoglycan chains, thereby inhibiting the third and final stage of bacterial cell wall synthesis. Subsequently, bacterial cell wall autolysins (such as autolysins) mediate cell lysis; carbenicillin may interfere with the action of autolysin inhibitors.
Penicillin and its metabolites are potent immunogens because they can bind to proteins and act as haptens to trigger an acute antibody-mediated immune response. The most common (approximately 95%) or "primary" determinant of penicillin allergy is the penicillin acyl determinant, which is produced after the opening of the penicillin β-lactam ring. This allows penicillin to bind to proteins via the amide group. "Secondary" determinants (occurring less frequently) are other metabolites, including native penicillin and penicillinic acid. /Penicillin/
A bactericidal agent; inhibits bacterial cell wall synthesis. Its action depends on penicillin's ability to reach and bind to penicillin-binding proteins (PBPs) located on the inner membrane of the bacterial cell wall. Penicillin-binding proteins (including transpeptidase, carboxypeptidase, and endopeptidase) are enzymes involved in the final stages of bacterial cell wall assembly and in the remodeling of the cell wall during bacterial growth and division. Penicillin binds to and inactivates penicillin-binding proteins, leading to a weakened bacterial cell wall and eventual lysis. /Penicillin/

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Therapeutic Uses
Penicillin
The main advantage of this drug is that it can usually cure serious infections caused by Pseudomonas spp. (especially Pseudomonas aeruginosa), ampicillin-resistant Proteus strains, and certain other Gram-negative bacteria.
Carbenicillin (injection) is indicated for the treatment of bacterial pneumonia caused by susceptible bacteria. (Included in the US product label)
Carbenicillin (injection) is indicated for the treatment of bone and joint infections caused by susceptible bacteria. /Included in the US product label/
For more complete data on the therapeutic uses of carbenicillin (14 types), please visit the HSDB record page.

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Drug Warnings
In rare cases, due to the risk of death, future use of penicillin must be prohibited, and patients should be warned. It must be emphasized again that even taking very small doses of this antibiotic can cause fatal allergic reactions.Penicillin Class
These drugs are less effective against penicillin G-sensitive microorganisms and are ineffective against Gram-negative bacteria. Microorganisms can gradually develop resistance, and cross-resistance to all penicillin classes is usually complete. Penicillin Classes
Because insufficient drug concentrations can lead to treatment failure, carbenicillin indole sodium should not be used when rapid increases in blood and/or urine concentrations are required, or in patients with severe renal impairment (i.e., creatinine clearance less than 10 ml/min). Carbenicillin indole sodium
Penicillin classes are excreted into breast milk at low concentrations. Although there are no documented serious problems in humans, penicillin use in breastfeeding women may cause allergies, diarrhea, candidiasis, and rashes in infants. /Penicillin/
For more complete data on drug warnings for carbenicillin (24 in total), please visit the HSDB records page.
Pharmacodynamics
Carbenicillin is a semi-synthetic penicillin. Although carbenicillin exhibits significant in vitro activity against a variety of Gram-positive and Gram-negative bacteria, its most important characteristic lies in its anti-pseudomonal and anti-protein activity. Due to the high concentration of carbapenem in urine after administration, carbapenem has been shown to be clinically effective against urinary tract infections caused by the following susceptible strains: Escherichia coli, Proteus mirabilis, Proteus vulgaris, Morganella Morganella, Pseudomonas spp., Providens retardans, Enterobacter spp., and Enterococcus faecalis (Enterococcus faecalis).

C17H18N2O6S

378.399623394012

378.089

C, 53.96; H, 4.79; N, 7.40; O, 25.37; S, 8.47

4697-36-3

Carbenicillin disodium;4800-94-6

20824

Typically exists as solid at room temperature

1.53g/cm3

737.8ºC at 760mmHg

400ºC

1.675

0.815

3

7

5

26

645

3

O=C([C@@H](C(C)(C)S[C@]1([H])[C@@H]2NC(C(C(O)=O)C3=CC=CC=C3)=O)N1C2=O)O

FPPNZSSZRUTDAP-UWFZAAFLSA-N

InChI=1S/C17H18N2O6S/c1-17(2)11(16(24)25)19-13(21)10(14(19)26-17)18-12(20)9(15(22)23)8-6-4-3-5-7-8/h3-7,9-11,14H,1-2H3,(H,18,20)(H,22,23)(H,24,25)/t9?,10-,11+,14-/m1/s1

(2S,5R,6R)-6-[(2-carboxy-2-phenylacetyl)amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid

Carboxybenzylpenicillin; 4697-36-3; alpha-Carboxybenzylpencillin; Carbenicilina; Carbenicilline; Carbenicillinum; Carboxybenzylpenicillin acid;

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.)