| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| 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 |
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 |
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 [11]
|
| References | |
| 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, inhibiting protein synthesis[11] - Indications: - Experimental: Pulmonary fibrosis[1] - Approved: Bacterial infections (off-label use)[11] - Regulatory status: Not yet approved for fibrotic diseases; further clinical validation is required[1] |
| Molecular Formula |
C22H22N2O8
|
|---|---|
| Molecular Weight |
442.42
|
| Elemental Analysis |
C, 59.73; H, 5.01; N, 6.33; O, 28.93
|
| CAS # |
914-00-1
|
| Related CAS # |
Methacycline hydrochloride;3963-95-9
|
| PubChem CID |
54675785
|
| Appearance |
Typically exists as solids at room temperature
|
| Density |
1.66g/cm3
|
| Boiling Point |
695.1ºC at 760mmHg
|
| Melting Point |
>150°C (dec.) (lit.)
|
| Flash Point |
374.2ºC
|
| Index of Refraction |
1.753
|
| LogP |
0.262
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
32
|
| Complexity |
998
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
O[C@]12C(O)=C3C(C4C(O)=CC=CC=4C([C@H]3[C@H](O)[C@H]1[C@@H](C(=C(C2=O)C(=O)N)O)N(C)C)=C)=O
|
| Synonyms |
methacycline; Metacycline; 914-00-1; Rondomycin; Methylenecycline;
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
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
|
|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (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.
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