Mean MIC: 125 ng/mL (E. coli)[1] MIC50: 1 mg/mL (A. baumannii)[2] MIC90: 2 mg/mL (A. baumannii)[2]
The primary target of Tigecycline tetramesylate is the 30S ribosomal subunit of bacteria. Its mechanism of action is similar to other tetracyclines: it binds to the 30S ribosomal subunit and inhibits bacterial protein synthesis by blocking the entry of amino-acyl tRNA molecules into the A-site of the ribosome, thereby preventing the incorporation of new amino acids into the growing peptide chain. However, due to its unique substitution at the 9-position of the minocycline backbone, it is able to bind to the ribosome with 5-fold higher affinity than tetracycline and is not affected by common resistance mechanisms like tet(M) and tet(K).

With IC50s of 4.72±0.54 and 3.06±0.85 μM (freshly generated), tigecycline (0.63-30 µM, preincubated for 4 days, administered for 72 hours) inhibits AML2 cells and HL-60 cells. After one day of preincubation, tigecycline inhibits HL-60 and AML2 cells with IC50 values of 4.27±0.45 and 5.64±0.55 μM, respectively. 60 cells exhibited 3.95±0.39 μM and 4.09±0.41 μM IC50s (three days preincubation). Tigecycline reduced its capacity to kill TEX human leukemia cells after 4 days of preincubation in saline, as shown by the CellTiter Flour assay. IC50~5 µM when freshly synthesized to IC50 >50 µM after 4 days of preincubation[1].

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In vitro studies demonstrate that Tigecycline tetramesylate is a broad-spectrum antibiotic with potent activity against Gram-positive, Gram-negative, and anaerobic bacteria. It has shown a mean inhibitory concentration (MIC) of approximately 125 ng/mL against E. coli (MG1655 strain) [41L11]. It exhibits potent activity against multidrug-resistant (MDR) Gram-positive cocci, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Its unique activity against MDR strains is due to its ability to overcome common tetracycline resistance mechanisms. Tigecycline has also been shown to reduce the capacity to kill human leukemia cells.

In NOD/SCID mice, tigecycline (50 mg/kg) administered intraperitoneally twice a day for 11 days decreases tumor mass and volume[1]. Tigecycline in saline has the following values: peak plasma concentration (Cmax), terminal half-life (t1/2), area under the plasma concentration-time curve (AUC), clearance (CL), and volume of distribution (Vz), in that order: 22.8μg/mL, 108.9 min, 1912.2min*μg/mL, 26.1 mL/min/kg, and 4109.4 mL/kg. For Tigecycline in formulation (60 mg/mL pyruvate, 3 mg/mL ascorbic acid, pH 7 in saline), the peak plasma concentration (Cmax), the terminal half-life (t1/2), the area under the plasma concentration-time curve (AUC), the clearance (CL), and the volume of distribution (Vz) are 15.7μg/mL, 110.3 min, 2036.5 min*μg/mL, 24.6 mL/min/kg, and 3906.2 mL/kg, respectively.

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In vivo activity of tigecycline has been extensively studied in animal models of infection and confirmed in clinical practice. It is an FDA-approved and commercially available antibiotic. In preclinical models of septicemia, skin infection, and intra-abdominal infections, tigecycline treatment has been shown to significantly reduce bacterial burden and increase survival rates in treated animals. Its in vivo efficacy is a direct result of its potent in vitro activity and its favorable tissue distribution, which leads to high concentrations in the tissues, including the skin and gastrointestinal tract.

The binding of tigecycline to the bacterial ribosome can be studied in a cell-free assay. A standard protocol involves using isolated 70S ribosomes from a susceptible bacterial strain (e.g., E. coli). The ribosomes are incubated with a fixed concentration of a radiolabeled tetracycline (e.g., [3H]-tetracycline) and increasing concentrations of unlabeled tigecycline. After incubation at 37degC for 30 minutes, the ribosomes are filtered through a nitrocellulose filter and washed with buffer. The filters are then placed in a scintillation vial, and the bound radioactivity is measured. A decrease in signal indicates that tigecycline is displacing the radiolabeled tetracycline, providing a direct measurement of its binding affinity.

Cell Viability Assay[1]
Cell Types: Human leukemic OCI-AML2, HL-60(ATCC) and TEX cell lines
Tested Concentrations: 0.63-30 µM
Incubation Duration: Preincubated for 4 days, treated for 72 hrs (hours)
Experimental Results: Inhibited AML2 cells and HL-60 cells with IC50s of 4.72±0.54 and 3.06±0.85 μM(freshly prepared).

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For standard susceptibility testing, the broth microdilution method is used. A stock solution of tigecycline tetramesylate is prepared in sterile water or DMSO. Two-fold serial dilutions of the antibiotic are made in cation-adjusted Mueller-Hinton broth (CAMHB) in a 96-well plate. A bacterial inoculum equivalent to a 0.5 McFarland standard (approximately 1.5 x 10^8 CFU/mL) is then added to each well, achieving a final concentration of approximately 5 x 10^5 CFU/mL. The plates are incubated at 35degC for 16-20 hours. The MIC is read as the lowest concentration of the antibiotic that results in no visible bacterial growth. The tetramesylate salt form of tigecycline ensures it is readily soluble in the growth medium.

Animal/Disease Models: NOD/SCID (severe combined immunodeficient) mouse with OCI-AML2 acute myeloid leukemia (AML) xenograft model[1]
Doses: 50 mg/kg
Route of Administration: intraperitoneal (ip)injection; twice a day; for 11 days
Experimental Results: decreased tumor volume and weight .

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Animal/Disease Models: NOD/SCID (severe combined immunodeficient) mouse[1]
Doses: 50 mg/kg
Route of Administration: intraperitoneal (ip)injection; 360 minutes
Experimental Results: The peak plasma concentration (Cmax), the terminal half-life (t1/2), area under the plasma concentration -time curve (AUC), clearance (CL) and volume of distribution (Vz) are 22.8 μg/mL, 108.9 min, 1912.2 min*μg/mL, 26.1 mL/min/kg, 4109.4 mL/kg, respectively.

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The efficacy of tigecycline in vivo is typically evaluated in a murine thigh infection model. Immunocompromised (neutropenic) mice are infected intramuscularly in the thigh with a bacterial suspension (e.g., S. aureus, E. coli). Two hours post-infection, the mice are treated subcutaneously with tigecycline tetramesylate at various doses (e.g., 1, 4, 16, and 64 mg/kg/day), administered in split doses (e.g., every 6 hours). After 24 hours of treatment, the mice are euthanized, and the thighs are harvested and homogenized. The bacterial load (CFU/thigh) is determined by plating serial dilutions of the homogenates on agar plates. The reduction in bacterial load compared to untreated control mice is used to calculate the static dose and the dose required to reduce bacterial counts by 1 log (ED50).

The pharmacokinetic properties of tigecycline are well-characterized. It is not significantly metabolized and is primarily excreted unchanged in the bile. The tetramesylate salt is used to ensure high water solubility for intravenous administration, which is the standard clinical route. Tigecycline has a large volume of distribution, indicating extensive tissue penetration, and its half-life is relatively long for an antibiotic, allowing for twice-daily dosing in the clinic. In research, the tetramesylate salt is a stable and soluble form for preparing solutions for in vivo (animal) administration. The compound is typically stored at 4degC, sealed, and away from moisture to maintain stability [41L4-L5].

The toxicity of tigecycline tetramesylate has been extensively studied in preclinical models and clinical trials. The most common adverse effects are gastrointestinal (nausea, vomiting, diarrhea, abdominal pain). A major safety concern is a higher risk of all-cause mortality observed in patients treated with tigecycline compared to other antibiotics, particularly in those with severe infections, which led to a "black box" warning from the FDA against its use in hospital-acquired pneumonia. It is generally avoided in pediatric patients and pregnant women due to potential effects on bone and tooth development. As a research chemical, it is for research use only.

[1]. Jitkova Y, et al. A novel formulation of tigecycline has enhanced stability and sustained antibacterial and antileukemic activity. PLoS One. 2014 May 28;9(5):e95281.
[2]. Falagas ME, et al. Activity of TP-6076 against carbapenem-resistant Acinetobacter baumannii isolates collected from inpatients in Greek hospitals. Int J Antimicrob Agents. 2018 Aug;52(2):269-271.

Tigecycline is a tetracycline derivative with its 5-hydroxyl and 6-methyl groups replaced by hydrogen atoms, and its 7- and 9-positions replaced by dimethylamino and (N-tert-butylglycyl)amino groups, respectively. It is a glycylcycline antibiotic with antibacterial activity against a variety of Gram-positive and Gram-negative bacteria, including tetracycline-resistant bacteria. It is used intravenously to treat complicated skin and soft tissue infections caused by susceptible bacteria. It is an antibacterial drug. It belongs to the tetracycline class of compounds and is a tert-α-hydroxy ketone. It is the conjugate base of tigecycline (1+). Tigecycline is a tetracycline antibacterial drug. It is a tetracycline derivative and can act as a protein synthesis inhibitor. It is used as a systemic antibacterial drug to treat complicated skin and intra-abdominal infections. Tigecycline is also used to treat community-acquired pneumonia. Tigecycline is a broad-spectrum, first-in-class glycylcycline antibiotic currently used to treat complicated skin infections, intra-abdominal infections, and community-acquired pneumonia. Furthermore, we have demonstrated that tigecycline possesses in vitro and in vivo anti-acute myeloid leukemia (AML) activity, attributed to its ability to inhibit mitochondrial translation. Tigecycline is relatively unstable after reconstitution, and this instability may limit its application in outpatient infusion therapy for infections and may hinder the development of optimal dosing regimens for AML. This study aimed to find a formulation that could improve the stability of the drug after reconstitution while maintaining its antibacterial and antileukemic activities. We tested a range of chemical additives to screen for excipients that could improve the stability of tigecycline solutions at room temperature for up to one week. We discovered a novel formulation containing the oxygen reducing agent ascorbic acid (3 mg/mL) and pyruvate (60 mg/mL) in physiological saline at pH 7.0, in which tigecycline (1 mg/mL) remains intact for at least 7 days under light-protected conditions. This formulation also maintained the drug's antibacterial and antileukemic activities in vitro. Furthermore, this novel formulation also preserved the antileukemic activity of tigecycline in vivo. Therefore, we identified and characterized a novel tigecycline formulation that retains its stability and efficacy after reconstitution. [1]

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Tigecycline tetramesylate is the mesylate salt form of the FDA-approved, broad-spectrum glycylcycline antibiotic, tigecycline. Its chemical name is GAR-936 tetramesylate. It is a research-grade chemical for antibiotic discovery and development [41L3-L4]. Tigecycline is a potent inhibitor of the bacterial 30S ribosome, and its tetramesylate salt form ensures excellent water solubility and chemical stability for both in vitro and in vivo research applications. It is strictly for research use and is not a commercial drug for human use. As a research tool, it is invaluable for studying bacterial protein synthesis and resistance mechanisms. It is typically stored at 4degC, away from moisture, to maintain its stability [41L4-L5].

C33H55N5O20S4

969.2323

Tigecycline;220620-09-7;Tigecycline hydrochloride;197654-04-9;Tigecycline mesylate;1135871-27-0;Tigecycline hydrate;1229002-07-6

137628652

Typically exists as Light yellow to yellow solid at room temperature

IBOQJGSRFKTAPT-LFSRUXGMSA-N

InChI=1S/C29H39N5O8.4CH4O3S/c1-28(2,3)31-11-17(35)32-15-10-16(33(4)5)13-8-12-9-14-21(34(6)7)24(38)20(27(30)41)26(40)29(14,42)25(39)18(12)23(37)19(13)22(15)36;4*1-5(2,3)4/h10,12,14,21,31,36-37,40,42H,8-9,11H2,1-7H3,(H2,30,41)(H,32,35);4*1H3,(H,2,3,4)/t12-,14-,21-,29-;;;;/m0..../s1

(4S,4aS,5aR,12aR)-9-[[2-(tert-butylamino)acetyl]amino]-4,7-bis(dimethylamino)-1,10,11,12a-tetrahydroxy-3,12-dioxo-4a,5,5a,6-tetrahydro-4H-tetracene-2-carboxamide;methanesulfonic acid

Tigecycline tetramesylate; GAR-936 tetramesylate; Tigecycline (tetramesylate);

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 :~100 mg/mL (~103.09 mM)
H2O :~50 mg/mL (~51.54 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.)