| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
RAV-2 reverse transcriptase (IC50 = 3.8 μg/mL)
HIV-1 reverse transcriptase (IC50 = 45 μg/mL) RSV reverse transcriptase (IC50 = 100 μg/mL) E. coli DNA polymerase I (IC50 = 230 μg/mL) [1] |
|---|---|
| ln Vitro |
Inhibition of RSV focus formation: AC inhibited focus formation by Rous sarcoma virus in chicken embryo fibroblast cells with an IC50 of 8.5 μg/mL. The CC50 for CEF cell growth was 180 μg/mL, giving a CC50/IC50 ratio of 21.2. [1]
Inhibition of RSV production: At a concentration of 62.5 μg/mL, AC inhibited RSV production by 93% during 0-24 hours post-infection and by 99% during 24-48 hours post-infection. At 31.3 μg/mL, inhibition was 75% and 92.7% respectively. [1] Anti-HIV activity in MT-4 cells: AC inhibited HIV-1-induced cytopathogenicity with an IC50 of 19.0 μg/mL. The CC50 in uninfected MT-4 cells was 79.0 μg/mL, giving a CC50/IC50 ratio of 4.2. [1] Anti-HIV activity in Sup-T1 cells: At 50 μg/mL, AC inhibited HIV-1 cytopathogenicity by 37% . The CC50 in uninfected Sup-T1 cells was 64.5 μg/mL. [1] Direct effect on virus particles: Incubation of RSV with 31.3 μg/mL AC at 4°C for 3 hours, followed by dilution to non-inhibitory concentrations, showed no appreciable direct inactivating effect on the virus. [1] |
| ln Vivo |
In RSV-infected chickens (3 days old), subcutaneous administration of AC at 200 mg/kg per day delayed tumor development on the wing web by up to 6 days. The mean survival time of control chickens (without treatment) was 14.0 ± 1.4 days, while treatment with AC prolonged survival time to 18.5 ± 2.4 days (P values <0.01, <0.05, <0.001 at various time points). Lower doses (50, 12.5, 3.13 mg/kg) also delayed tumor formation but to a lesser extent. [1]
|
| Enzyme Assay |
RAV-2 reverse transcriptase assay: The reaction mixture (50 μL) contained 50 mM Tris-HCl (pH 8.3), 50 mM KCl, 10 mM MgCl₂, 3 mM dithiothreitol, 0.5 μg poly(rA)-pdT12-18, 10 μM [³H]TTP (30 GBq/mmol), and 0.1 U of RAV-2 RT. Test compounds were added at various concentrations. The reaction was incubated at 37°C for 30 min, stopped by cooling to 0°C and addition of 20 μL of 0.1 M EDTA (pH 8.0). Incorporation of [³H]TTP into polymers was determined using DE81 ion exchange filter papers. All test compounds were dissolved in DMSO, with a final DMSO concentration of 5% in the reaction mixture. [1]
HIV-1 reverse transcriptase assay: Performed using HIV-1 RT prepared from an E. coli expression system. The assay conditions followed a previously reported method. [1] RSV reverse transcriptase assay: The reaction mixture (100 μL) contained 50 mM Tris-HCl (pH 8.1), 50 mM NaCl, 3 mM DTT, 5 mM MgCl₂, 100 μM [³H]TTP (37 GBq/mmol), 1% Triton X-100, 2 μg poly(rA)-dT15, and 30 μL of RSV (2.3×10⁶ focus forming units/mL). The reaction was incubated at 37°C for 1 hour. Duplicate 45 μL aliquots were spotted onto filter papers, and trichloroacetic acid-insoluble radioactivity was measured. [1] E. coli DNA polymerase I assay: Performed according to a referenced method (Hanajima et al., 1985). [1] Time-of-addition experiment: AC (4 μg/mL) was added at various time points after initiation of the RAV-2 RT reaction. The reaction remained linear over 30 min, and AC inhibited DNA synthesis at all times after reaction onset, indicating that AC can exert inhibition even after enzyme-template-primer-substrate complex formation. [1] |
| Cell Assay |
Cytotoxicity assay on chicken embryo fibroblast cells: CEF cells were seeded at 8.75×10⁴ cells/0.1 mL/well in a 96-well microplate and incubated with various concentrations of test compounds at 37°C for 6 hours in 5% CO₂. Medium was removed, cells washed with PBS, then 90 μL of medium (Ham's F-10 with 5% FBS and 1% chicken serum) and 10 μL of test compound solution (in water or DMSO) were added. Color blank wells containing diluted compound solutions without cells were prepared. After 4 days of incubation, viable CEF cells were measured by the MTT method. [1]
RSV focus formation assay: CEF cells (3.5×10⁵ cells/1 mL/well) were incubated in a 24-well microplate at 37°C for 6 hours. Cells were infected with RSV at a multiplicity of infection of 0.0001 for 2 hours in the presence of test compounds. Focus assays were performed as described previously (Vogt, 1969). Each experimental value was obtained from the average of four replicates, and experiments were repeated at least twice. [1] RSV production inhibition assay: CEF cells were infected with RSV (MOI 0.01) simultaneously with addition of test compounds. After 2 hours, virus was removed and fresh medium containing test compounds was added. Infected cells were exposed to test compounds for 24 hours, then first samples were taken for focus assay. Medium was then changed to fresh medium without test compounds, and cells were further incubated. Second samples were taken at 48 hours post-infection. Test compounds in the virus solution were sufficiently diluted to have no influence on the focus assay. [1] Anti-HIV assay in MT-4 and Sup-T1 cells: The procedure for measuring anti-HIV activity in MT-4 cells followed a previously described method (Pauwels et al., 1988). HIV-1 solution adjusted to 20 TCID₅₀ per well was used. Anti-HIV activities and cytotoxicities of compounds with Sup-T1 cells were determined by a similar procedure at day 7 post-infection. [1] |
| Animal Protocol |
Three-day-old SPAFAS chickens were used. AC was dissolved in saline, and the pH was adjusted to 7.4 using NaOH solution. Each chicken was inoculated subcutaneously in the wing web with 0.1 mL of virus solution (1×10³ FFU/mL) containing various concentrations of AC. AC was administered subcutaneously consecutively for 6 days after virus inoculation. Dosages tested were 200, 50, 12.5, and 3.13 mg/kg per day. Tumor size was measured once daily from day 1 to day 14 post-infection, and mortalities were recorded. For the control group, no AC treatment was given. In uninfected chickens, the same AC treatment (200 mg/kg per day) did not cause any deleterious effect. [1]
|
| Toxicity/Toxicokinetics |
In vitro cytotoxicity: The CC₅₀ (50% cytotoxic concentration) of AC in CEF cells was 180 μg/mL; in MT-4 cells it was 79.0 μg/mL; in Sup-T1 cells it was 64.5 μg/mL. [1]
In vivo toxicity: Administration of AC at 200 mg/kg per day to uninfected chickens did not cause any deleterious effect. [1] |
| References |
|
| Additional Infomation |
Alizarin complex is a dihydroxyanthraquinone compound with hydroxyl groups at C-1 and C-2 positions and a bis[(carboxymethyl)amino]methyl substituent at the 3 position. It is a colorimetric reagent. It is a dicarboxylic acid and a dihydroxyanthraquinone.
AC is an anthraquinone containing hydroxy groups at positions corresponding to R₅ and R₆, which appear important for activity against RAV-2 RT. Although AC is a chelating reagent for metal ions (e.g., cerium(III) used for fluoride determination), its inhibitory effect on RAV-2 RT is not due to chelation of Mg²⁺, as increasing Mg²⁺ concentration up to 100 μM did not influence inhibition. [1] The inhibitory concentration of AC on RSV focus formation (8.5 μg/mL) was approximately one-tenth of that required to inhibit RSV RT (100 μg/mL), suggesting that the anti-RSV effect may not be entirely due to RT inhibition. Similarly, anti-HIV-1 IC₅₀ in MT-4 cells (19 μg/mL) was lower than the IC₅₀ for HIV-1 RT (45 μg/mL), indicating additional mechanisms. AC only affected tumor induction when administered simultaneously with virus; if administration was delayed until 24 hours after virus inoculation, no effect was observed. Once a tumor appeared, daily AC did not influence tumor growth rate, consistent with AC interfering with the viral replicative cycle rather than tumor growth itself. [1] AC may act similarly to hypericin (a dimeric anthraquinone that inhibits viral assembly). The exact mechanism of anti-HIV-1 action remains to be elucidated. AC is considered a lead compound for developing more potent and selective anti-HIV agents. [1] |
| Molecular Formula |
C19H15NO8
|
|---|---|
| Molecular Weight |
385.3243
|
| Exact Mass |
385.079
|
| CAS # |
3952-78-1
|
| PubChem CID |
65132
|
| Appearance |
Yellow to brown solid powder
|
| Density |
1.6±0.1 g/cm3
|
| Boiling Point |
687.1±55.0 °C at 760 mmHg
|
| Melting Point |
~185 °C (dec.)
|
| Flash Point |
369.4±31.5 °C
|
| Vapour Pressure |
0.0±2.2 mmHg at 25°C
|
| Index of Refraction |
1.720
|
| LogP |
3.38
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
28
|
| Complexity |
648
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
PWIGYBONXWGOQE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H15NO8/c21-13(22)7-20(8-14(23)24)6-9-5-12-15(19(28)16(9)25)18(27)11-4-2-1-3-10(11)17(12)26/h1-5,25,28H,6-8H2,(H,21,22)(H,23,24)
|
| Chemical Name |
2-[carboxymethyl-[(3,4-dihydroxy-9,10-dioxoanthracen-2-yl)methyl]amino]acetic acid
|
| 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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), 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)
|
| Solubility (In Vitro) |
DMSO : ≥ 100 mg/mL (~259.52 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.49 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% 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 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O 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 | 2.5952 mL | 12.9762 mL | 25.9525 mL | |
| 5 mM | 0.5190 mL | 2.5952 mL | 5.1905 mL | |
| 10 mM | 0.2595 mL | 1.2976 mL | 2.5952 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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