| Size | Price | |
|---|---|---|
| 1g | ||
| Other Sizes |
| Targets |
Tetracycline-Responsive Transcriptional Regulator (TetR): Binds to TetR to modulate gene expression in inducible systems
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|---|---|
| ln Vitro |
1. Gene Expression Regulation in NIH 3T3 Cells
- Cell Line: NIH 3T3 cells stably expressing HER2 under Tet-Off or Tet-On system control.
- Treatment:
- Tet-Off system: 4-Epidoxycycline (10 ng/mL) for 24 hours to repress HER2 expression.
- Tet-On system: 4-Epidoxycycline (10 ng/mL) for 48 hours to induce HER2 expression.
- Results:
- Repressed HER2 protein levels by 80% in Tet-Off cells (Western blot) [1]
- Induced HER2 protein levels by 5-fold in Tet-On cells (Western blot) [1] |
| ln Vivo |
1. Tumor Regression in HER2-Expressing Mouse Model
- Animal Model: Nude mice bearing NIH 3T3-HER2 xenografts (tumor volume ~1.6 cm³).
- Treatment:
- Group 1: 4-Epidoxycycline (7.5 mg/mL in drinking water, ad libitum) [1]
- Group 2: 4-Epidoxycycline (10 mg/kg, subcutaneous injection, daily) [1] - Duration: 7 days. - Results: - Both treatments induced >95% tumor regression (measured by caliper) [1] - No significant difference in efficacy compared to doxycycline (7.5 mg/mL in drinking water) [1] |
| Enzyme Assay |
1. Luciferase Reporter Assay for TetR Binding
- Reagents: HEK293 cells transfected with TetR-responsive luciferase reporter plasmid.
- Protocol:
1. Seed cells (5×10⁴ cells/well) in 24-well plates and incubate overnight [1]
2. Treat with 4-Epidoxycycline (0.1–100 ng/mL) for 24 hours [1] 3. Lyse cells and measure luciferase activity using a luminometer [1] - Results: 4-Epidoxycycline (10 ng/mL) induced luciferase activity by 10-fold in Tet-On cells [1] |
| Cell Assay |
1. Western Blot Analysis of HER2 Expression
- Cell Line: NIH 3T3-HER2 cells.
- Protocol:
1. Treat cells with 4-Epidoxycycline (10 ng/mL) for 24 hours [1]
2. Lyse cells in RIPA buffer and separate proteins by SDS-PAGE [1] 3. Transfer to PVDF membrane and probe with anti-HER2 antibody [1] - Results: HER2 protein levels were reduced by 80% in Tet-Off cells treated with 4-Epidoxycycline [1] |
| Animal Protocol |
1. Drinking Water Administration in Mice
- Animal Model: Nude mice (6–8 weeks old).
- Protocol:
1. Prepare 4-Epidoxycycline solution (7.5 mg/mL) in sterile water containing 0.1% sucrose [1]
2. Provide ad libitum access to medicated water for 7 days [1] 3. Monitor tumor volume every 2 days using calipers [1] 2. Subcutaneous Injection in Mice - Animal Model: Nude mice (6–8 weeks old). - Protocol: 1. Dissolve 4-Epidoxycycline (10 mg/kg) in sterile PBS [1] 2. Inject subcutaneously into the dorsal flank once daily for 7 days [1] 3. Monitor tumor volume every 2 days using calipers [1] |
| ADME/Pharmacokinetics |
Distribution: - 4-Epicocycline was detected in tumor tissue at concentrations comparable to plasma concentrations (determined by high performance liquid chromatography) [1] - Excretion: - No specific data have been reported in the literature [1] #### Toxicity/Toxicokinetics - In vivo safety - 4-Epicocycline (7.5 mg/mL in drinking water) did not cause diarrhea or weight loss in mice [1] - No significant changes were observed in liver and kidney function indicators (ALT, AST, BUN) compared to the solvent control group [1]
|
| Toxicity/Toxicokinetics |
- In vivo safety - Addition of 4-epicocyclone (7.5 mg/mL) to drinking water did not cause diarrhea or weight loss in mice [1]
- No significant changes were observed in liver and kidney function indicators (ALT, AST, BUN) compared to the solvent control group [1] |
| References | |
| Additional Infomation |
- Background: - 4-Epicocycline is a liver metabolite of doxycycline, and its antibacterial activity is reduced [1]
- Advantages: - It avoids intestinal flora dysbiosis compared to doxycycline [1] - Mechanism of action: - It binds to TetR and induces or inhibits gene expression in the Tet-On/Tet-Off system [1] |
| Molecular Formula |
C22H24N2O8
|
|---|---|
| Molecular Weight |
444.43
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| Exact Mass |
444.153
|
| CAS # |
6543-77-7
|
| PubChem CID |
54676637
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| Appearance |
Typically exists as solids at room temperature
|
| Melting Point |
>176?C (dec.)
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| LogP |
0.353
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
32
|
| Complexity |
956
|
| Defined Atom Stereocenter Count |
6
|
| SMILES |
CC1C2C(C3C(C(=O)C(=C(C3(C(=O)C2=C(C4=C1C=CC=C4O)O)O)O)C(=O)N)N(C)C)O
|
| InChi Key |
SGKRLCUYIXIAHR-NLJUDYQYSA-N
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| InChi Code |
InChI=1S/C22H24N2O8/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/h4-7,10,14-15,17,25-27,30,32H,1-3H3,(H2,23,31)/t7-,10+,14+,15+,17-,22-/m0/s1
|
| Chemical Name |
(4R,4aR,5S,5aR,6R,12aR)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4a,5,5a,6-tetrahydro-4H-tetracene-2-carboxamide
|
| Synonyms |
4-Epi-doxycycline; 4-Epidoxycycline; 6543-77-7; 4-epi-Doxycycline; 6-Deoxy-4-epioxytetracycline; 4-Epi Doxycycline (>70%); TR57F2W7Q6; (4R,4aR,5S,5aR,6R,12aR)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4a,5,5a,6-tetrahydro-4H-tetracene-2-carboxamide; 2-Naphthacenecarboxamide, 4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-, (4R,4aR,5S,5aR,6R,12aS)-;
|
| 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 |
| 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) |
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
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|---|---|
| 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 | 2.2501 mL | 11.2504 mL | 22.5007 mL | |
| 5 mM | 0.4500 mL | 2.2501 mL | 4.5001 mL | |
| 10 mM | 0.2250 mL | 1.1250 mL | 2.2501 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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