| Size | Price | Stock | Qty |
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| 250mg |
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| Targets |
- TGF-β1-Induced Non-Smad Signaling Pathways: Inhibits c-Jun N-terminal kinase (JNK), p38, and Akt activation with an IC50 of ~5 μM in A549 cells [1]
- Bacterial Ribosomal 30S Subunit: Binds to 30S ribosomal subunit to inhibit protein synthesis (no IC50 reported) |
|---|---|
| ln Vitro |
Meclocycline hydrochloride (10, 20 μM; for 48 hours) inhibits the α-smooth muscle that TGF-β1 induces in primary alveolar epithelial cells. It reacts to TGF-β1-induced non-Smad buffering, such as c-Jun N-telogen, p38, and Akt activation, but it does not inhibit the reactivation of Smad or β-catenin. -Jun N had no influence on lung blast fibroblasts' TGF-β1 response, p38 or Akt activation, or termination of cleavage [1].
1. EMT Inhibition in A549 Cells - Cell Line: A549 human alveolar epithelial cells treated with TGF-β1 (5 ng/mL). - Treatment: Methacycline (1–20 μM) for 48 hours. - Assays: - Immunofluorescence: Reduced E-cadherin loss and fibronectin accumulation [1] - qPCR: Downregulated α-SMA, Snail1, and collagen I mRNA expression [1] - Transwell Migration: Inhibited TGF-β1-induced cell migration by 40–60% at 20 μM [1] 2. Non-Smad Pathway Modulation - Western Blot: Blocked TGF-β1-induced phosphorylation of JNK (Thr183/Tyr185), p38 (Thr180/Tyr182), and Akt (Ser473) in A549 cells [1] - Reporter Assay: No effect on Smad or β-catenin transcriptional activity [1] |
| ln Vivo |
In animal, meclocycline hydrochloride (100 mg/kg/day; intraperitoneally; starting 10 days after intratracheal bleomycin) raises newborn rates at day 17. Typical EMT markers generated by bleomycin, Snail1, Twist1, collagen I, as well as fibrinogen and mRNA, were fragmented by melocycline hydrochloride (day 17) [1].
1. Bleomycin-Induced Pulmonary Fibrosis in Mice - Animal Model: C57BL/6 mice intratracheally instilled with bleomycin (2 U/kg). - Treatment: - Methacycline (100 mg/kg/day) via intraperitoneal injection starting on day 10 post-bleomycin [1] - Vehicle control: DMSO/saline solution [1] - Results: - Survival: Improved survival rate at day 17 (p < 0.01) [1] - Histology: Reduced collagen deposition (Masson’s trichrome staining) and α-SMA-positive myofibroblasts in lung tissue [1] - mRNA Analysis: Downregulated Snail1, Twist1, collagen I, and fibronectin expression in lung homogenates [1] |
| Cell Assay |
1. EMT Phenotype Analysis in A549 Cells
- Protocol:
1. Seed A549 cells in 96-well plates and treat with TGF-β1 (5 ng/mL) + methacycline (1–20 μM) for 48 hours [1]
2. Fix cells with 4% paraformaldehyde and stain for E-cadherin (green) and fibronectin (red) using immunofluorescence [1] 3. Quantify fluorescence intensity using high-content imaging [1] - Results: Methacycline at 20 μM restored E-cadherin expression to 80% of baseline and reduced fibronectin by 60% [1] 2. Phosphorylation Profiling of JNK/p38/Akt - Protocol: 1. Treat A549 cells with TGF-β1 (5 ng/mL) + methacycline (10 μM) for 30 minutes [1] 2. Lyse cells and perform Western blot using phospho-specific antibodies for JNK, p38, and Akt [1] - Results: Complete inhibition of TGF-β1-induced phosphorylation of JNK, p38, and Akt [1] |
| Animal Protocol |
1. Bleomycin Model in Mice
- Animal Model: 8-week-old male C57BL/6 mice.
- Bleomycin Instillation:
1. Anesthetize mice and instill bleomycin (2 U/kg dissolved in 50 μL saline) intratracheally [1]
- Drug Administration: 1. From day 10 post-bleomycin, inject methacycline (100 mg/kg) dissolved in DMSO/saline (1:9 v/v) intraperitoneally daily for 7 days [1] - Sample Collection: 1. Sacrifice mice on day 17 and collect lung tissue for histology, RNA extraction, and protein analysis [1] |
| ADME/Pharmacokinetics |
Absorption: Oral bioavailability in rodents is moderate (approximately 30%) (not directly measured in this study) [9]
- Distribution: Accumulates in lung tissue (concentration 2-3 times higher than in plasma) [1] - Metabolism: Mainly metabolized by hepatic cytochrome P450 enzymes (not directly measured in this study) [9] - Excretion: Excreted by the kidneys (40-60% unchanged) and bile (20-30%) [9] - Half-life: Approximately 12-18 hours in rodents (not directly measured in this study) [9] |
| Toxicity/Toxicokinetics |
Acute toxicity: - Oral LD50 in mice > 2000 mg/kg (not directly measured in this study) [10] - Chronic toxicity: - No significant weight loss or liver and kidney dysfunction was observed in mice after 7 consecutive days of treatment with 100 mg/kg cycline [1] - Drug interactions: Chelates with divalent cations (e.g., Ca²⁺, Mg²⁺), reducing absorption
54685047 Oral LD50 in rats > 2 gm/kg, Japanese Pharmacopoeia, 6(818), 1982 54685047 Intraperitoneal LD50 in rats 252 mg/kg, Toxicology and Applied Pharmacology, 18(185), 1971 [PMID:5542824] 54685047 Intravenous LD50 in rats 202 mg/kg, Japanese Pharmacopoeia, 6(818), 1982 54685047 Intraperitoneal LD50 in mice: 288 mg/kg, Toxicology and Applied Pharmacology, 18(185), 1971 [PMID:5542824] 54685047 Intravenous LD50 in mice: 193 mg/kg, Japanese Medicine, 6(818), 1982 |
| References |
[1]. Ying Xi, et al. Inhibition of epithelial-to-mesenchymal transition and pulmonary fibrosis by methacycline. Am J Respir Cell Mol Biol. 2014 Jan;50(1):51-60.
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| Additional Infomation |
Background: - Methionine is a semi-synthetic tetracycline antibiotic that has been repurposed for antifibrotic therapy[1][11]
- Mechanism: - Antifibrotic: Inhibits TGF-β1-induced non-Smad signaling pathways (JNK/p38/Akt), thereby blocking EMT and fibroblast activation[1] - Antibacterial: Binds to bacterial 30S ribosomal subunits, thereby inhibiting protein synthesis[11] - Indications: - Experimental: Pulmonary fibrosis[1] - Approved: Bacterial infections (off-label use)[11] - Regulatory status: Not approved for fibrotic diseases; further clinical validation is required[1] According to state or federal labeling requirements, cycline hydrochloride may cause developmental toxicity. Cyclonine hydrochloride is an organic molecular entity. It is a broad-spectrum semi-synthetic antibiotic associated with tetracycline antibiotics, but it is excreted more slowly and maintains effective blood concentrations for a longer period of time. See also: Meclocycline hydrochloride (note moved to). |
| Molecular Formula |
C22H22N2O8
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|---|---|
| Molecular Weight |
442.4187
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| Exact Mass |
478.114
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| Elemental Analysis |
C, 55.18; H, 4.84; Cl, 7.40; N, 5.85; O, 26.73
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| CAS # |
3963-95-9
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| Related CAS # |
914-00-1; 41672-87-1 (trolamine); 3963-95-9 (HCl);
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| PubChem CID |
54685047
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| Appearance |
Typically exists as Light yellow to yellow solid at room temperature
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| Boiling Point |
813.8ºC at 760 mmHg
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| LogP |
1.064
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
33
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| Complexity |
998
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| Defined Atom Stereocenter Count |
5
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| SMILES |
O([H])[C@]12C(=C(C(N([H])[H])=O)C([C@]([H])([C@]1([H])[C@]([H])([C@]1([H])C(=C([H])[H])C3C([H])=C([H])C([H])=C(C=3C(=C1C2=O)O[H])O[H])O[H])N(C([H])([H])[H])C([H])([H])[H])=O)O[H]
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| InChi Key |
VXPSARQTYDZXAO-CCHMMTNSSA-N
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| InChi Code |
InChI=1S/C22H22N2O8.ClH/c1-7-8-5-4-6-9(25)11(8)16(26)12-10(7)17(27)14-15(24(2)3)18(28)13(21(23)31)20(30)22(14,32)19(12)29;/h4-6,10,14-15,17,25-27,30,32H,1H2,2-3H3,(H2,23,31);1H/t10-,14-,15+,17+,22+;/m1./s1
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| Chemical Name |
(4S,4aR,5S,5aR,12aR)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methylidene-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide;hydrochloride
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| Synonyms |
METHACYCLINE HYDROCHLORIDE; 3963-95-9; Methacycline HCl; Adriamicina; Ciclobiotic; Germiciclin; Metadomus; Globociclina;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~104.41 mM)
H2O : ~6.67 mg/mL (~13.93 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.22 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2603 mL | 11.3015 mL | 22.6030 mL | |
| 5 mM | 0.4521 mL | 2.2603 mL | 4.5206 mL | |
| 10 mM | 0.2260 mL | 1.1301 mL | 2.2603 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT02099240 | Terminated | Drug: oral antibiotics Procedure: intravenous antibiotics |
Osteomyelitis | Julio Ramirez | 2014-03-06 | Early Phase 1 |
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