| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary target of 4-epianhydrochlortetracycline hydrochloride is matrix metalloproteinase-9 (MMP-9/Gelatinase B). Unlike tetracycline, which primarily targets the bacterial 30S ribosome, this derivative exerts its biological effects through inhibition of MMPs, particularly gelatinase B (EC 3.4.24.35). The inhibition mechanism is thought to occur through chelation of the zinc ion at the enzyme's active site, a mechanism common to many tetracycline derivatives. This MMP inhibition leads to downstream effects including suppression of osteoclast differentiation by blocking MMP-9-mediated cleavage of histone H3, an event crucial for osteoclast formation.
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| ln Vitro |
In vitro studies demonstrate that 4-epianhydrochlortetracycline hydrochloride is a potent inhibitor of gelatinase B (MMP-9) in a dose-dependent manner. Quantitative inhibition data show 100% inhibition at 500 μg/mL, 70% inhibition at 125 μg/mL, and 40% inhibition at 31 μg/mL. At a concentration of 500 μg/mL, the compound completely inhibits neutrophil-derived MMP-9 degradation of denatured collagen. Unlike tetracycline, this derivative lacks significant antibiotic activity, although it retains some measurable inhibitory effects against certain bacterial strains including Shewanella (MIC: 2 mg/L), Escherichia coli (MIC: 1 mg/L), and Pseudomonas aeruginosa (MIC: 64 mg/L).
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| ln Vivo |
In vivo studies on the biological activity of 4-epianhydrochlortetracycline hydrochloride are limited, as the compound is primarily utilized as a reference standard and research tool rather than a therapeutic agent. However, its role as an MMP-9 inhibitor suggests potential applications in models of chronic inflammatory diseases where MMPs play a critical role in tissue remodeling and inflammation. The downstream effect of MMP-9 inhibition—suppression of osteoclast differentiation through blockade of histone H3 cleavage—indicates potential therapeutic relevance in bone-related disorders such as osteoporosis, though direct in vivo efficacy data for this specific derivative are currently limited.
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| Enzyme Assay |
Methodology for Gelatinase B (MMP-9) Inhibition Assay: The inhibitory activity of 4-epianhydrochlortetracycline hydrochloride on MMP-9 is measured using a microtiter assay for inhibitors as described by Paemen et al., 1996. Pro-MMP-9 is first activated to its catalytic form using p-aminophenylmercuric acetate (APMA). The activated MMP-9 is then pre-incubated with various concentrations of the test compound (e.g., 31, 125, 500 μg/mL) in an appropriate assay buffer. A fluorescently labeled gelatin substrate is added to the enzyme-inhibitor mixture. The increase in fluorescence, resulting from cleavage of the gelatin substrate by MMP-9, is monitored over time using a fluorescence plate reader. The rate of substrate cleavage is calculated for each inhibitor concentration, and the percentage of inhibition is determined by comparing the rates in the presence of the inhibitor to the rate of an uninhibited control.
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| Cell Assay |
Methodology for Antibacterial Activity Testing (MIC Determination): The minimum inhibitory concentration (MIC) of 4-epianhydrochlortetracycline hydrochloride against bacterial strains can be determined using standard broth microdilution methods. Bacterial suspensions are adjusted to 0.5 McFarland standard (~1.5 × 10⁸ CFU/mL) and diluted to achieve a final inoculum of ~5 × 10⁵ CFU/mL. Two-fold serial dilutions of the compound are prepared in 96-well plates (concentration range: 0.03-64 μg/mL or higher). Each well receives bacterial suspension and compound dilution, followed by incubation at 37°C for 18-24 hours. The MIC is defined as the lowest concentration with no visible bacterial growth. For reference, reported MIC values include: Shewanella (2 mg/L), Escherichia coli (1 mg/L), and Pseudomonas aeruginosa (64 mg/L).
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| Animal Protocol |
Methodology for Zebrafish Embryo Toxicity Assay: The toxicological profile of 4-epianhydrochlortetracycline hydrochloride has been characterized using a zebrafish embryo model. Zebrafish embryos are exposed to various concentrations of the compound (e.g., 5-20 mg/L) for up to 96 hours. Endpoints measured include: survival rate (for LC50 determination), hatching rate (to assess delayed hatching), heart rate (to assess cardiovascular toxicity), and morphological malformations (for EC50 determination). The study demonstrated dose-dependent lethal and developmental effects, with an LC50 of 29.13 mg/L and an EC50 (for malformation) of 8.57 mg/L after 96 hours of exposure. Oxidative stress markers including reactive oxygen species (ROS), malondialdehyde (MDA), and protein carbonyl (PC) content can also be measured to assess toxicity mechanisms.
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| ADME/Pharmacokinetics |
Comprehensive pharmacokinetic data for 4-epianhydrochlortetracycline hydrochloride specifically are limited in publicly available literature, as the compound is primarily used as a secondary standard and research tool rather than a therapeutic candidate. However, given its structural relationship to tetracycline and chlortetracycline degradation products, the compound is expected to exhibit properties similar to other tetracycline derivatives, including potential for tissue distribution, particularly to bone and teeth due to calcium chelation. The compound is hygroscopic and requires storage at -20°C under inert atmosphere to maintain stability. It shows solubility in DMF, DMSO, ethanol, methanol, and water. The compound can be used as a secondary standard for analytical method development and stability monitoring of tetracycline
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| Toxicity/Toxicokinetics |
According to GHS classification (Regulation EC No 1272/2008), 4-epianhydrochlortetracycline hydrochloride is classified as hazardous. Hazard statements include: H302 (Harmful if swallowed) - Acute Oral Toxicity Category 4; H319 (Causes serious eye irritation) - Eye Irritation Category 2; H361d (Suspected of damaging the unborn child) - Reproductive Toxicity Category 2. The precautionary signal word is "Warning". Zebrafish embryo studies have demonstrated dose-dependent toxicity with an LC50 of 29.13 mg/L and EC50 of 8.57 mg/L (for malformation) after 96 hours of exposure. The compound induces oxidative stress, evidenced by increased levels of reactive oxygen species (ROS), malondialdehyde (MDA), and protein carbonyl (PC) content, which is linked to the induction of cell apoptosis. No carcinogenic effects have been reported, and the compound is not listed as a carcinogen by IARC, NTP, or OSHA. Standard laboratory safety precautions, including eye protection and appropriate personal protective equipment, are recommended when handling this compound.
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| References |
| Molecular Formula |
C22H22CL2N2O7
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|---|---|
| Molecular Weight |
497.33
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| Exact Mass |
496.08040
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| CAS # |
158018-53-2
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| PubChem CID |
54710413
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| Appearance |
Typically exists as solids at room temperature
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| Boiling Point |
623ºC at 760 mmHg
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| Melting Point |
222ºC
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| Flash Point |
330.6ºC
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| Vapour Pressure |
2.21E-16mmHg at 25°C
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| LogP |
2.612
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
33
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| Complexity |
892
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=C2CC3C(C(=O)C(=C(C3(C(=O)C2=C(C4=C(C=CC(=C14)Cl)O)O)O)O)C(=O)N)N(C)C.Cl
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| InChi Key |
ISGAAFMBTIWTEU-PXAZKYFKSA-N
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| InChi Code |
InChI=1S/C22H21ClN2O7.ClH/c1-7-8-6-9-16(25(2)3)18(28)15(21(24)31)20(30)22(9,32)19(29)13(8)17(27)14-11(26)5-4-10(23)12(7)14;/h4-5,9,16,26-27,30,32H,6H2,1-3H3,(H2,24,31);1H/t9-,16+,22-;/m0./s1
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| Chemical Name |
(4R,4aS,12aR)-7-chloro-4-(dimethylamino)-1,10,11,12a-tetrahydroxy-6-methyl-3,12-dioxo-4a,5-dihydro-4H-tetracene-2-carboxamide;hydrochloride
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| Synonyms |
158018-53-2; 4-Epianhydrochlortetracycline Hydrochloride; 4-Epianhydrochlortetracycline (hydrochloride); (4R,4aS,12aR)-7-chloro-4-(dimethylamino)-1,10,11,12a-tetrahydroxy-6-methyl-3,12-dioxo-4a,5-dihydro-4H-tetracene-2-carboxamide;hydrochloride; 4-Epianhydrochlortetracycline hydrochloride, can be used as secondary standard;
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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 |
| 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 | 2.0107 mL | 10.0537 mL | 20.1074 mL | |
| 5 mM | 0.4021 mL | 2.0107 mL | 4.0215 mL | |
| 10 mM | 0.2011 mL | 1.0054 mL | 2.0107 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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