| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
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| Targets |
Antiviral; Epstein-Barr virus early antigen (EBV-EA); anticancer, insecticidal, nematocidal, anti-inflammatory
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|---|---|
| ln Vitro |
The osteoblasts treated with Azadirachtin B (1 pM-100 µM; 48 hours) proliferated most at doses of 10 nM and 100 pM [1]. Azadirachtin B enhanced RunX-2 expression by about 2.5-fold at 10 nM, ALP expression by about 2.8-fold at 10 nM and 100 pM, and OCN expression by about 2.5 at 10 nM in comparison to the control. times[1]. The diamondback moth (Plutella xylostella) is poisonous to azadirachtin B (Compound 4), with an LD50 of 4.85-1.06 µg/g body weight in 92 hours [2]. The Epstein-Barr virus early antigen (EBV-EA) is activated by tetradecanoylphorbol-13-acetate (TPA) and is moderately or potently inhibited by azadirachtin B (Compound 21) (IC50 384 molar ratio/32 pmol TPA) [3].
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| ln Vivo |
Compound 21 (oral) azadirachtin B was tested for its anti-tumor starting effect using mice skin tumors that were produced in two stages using TPA as a promoter and peroxynitrite (ONOO-; PN) as an initiator. According to the findings, there was a strong inhibitory activity [3].
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| Enzyme Assay |
Alkaline phosphatase activity (ALP assay): [1]
Osteoblast cells were seeded into 96 well culture plates and incubate to reach 50-70% confluency. After that, the medium was replaced and cells were incubated with differentiation medium for 48h at various concentrations of compounds. After 48h, media were replaced and washed the plates with PBS and freeze the plates into -800C. For ALP assay, we used our previous published protocols. Absorbance was measured at 405nm[1]. Mineralization assay: [1] Osteoblast cells were treated with compounds and cultured medium with α-MEM, containing 1% penicillin/streptomycin 50 µg/ml ascorbic acid, and 10 mM β-glycerophosphate at 370C. After 12 days of treatment, cells were fixed with 4% paraformaldehyde for 30 min and stained with 40mM alizarin red S stain. Calcium deposition nodules were stained red. Extraction of calcium precipitates was determined by measuring the absorbance at 405nM[1]. RNA preparation and reverse transcriptase polymerase chain reaction (RT-PCR): [1] The total cellular RNA was isolated using TRIzol reagent and RT-PCR was performed using RevertAidcDNA synthesis Kit using 2 µg total RNA in 20 µl reaction volume. GAPDH was used as internal control (housekeeping gene). The design of sense and antisense oligonucleotide primers was based on published cDNA sequences using the Universal ProbeLibrary . For Q-PCR, the cDNA was amplified using Light Cycler 480. Primers are described in table 1. |
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: Osteoblasts Tested Concentrations: 1 pM, 100 pM, 10 nM, 1 µM, 100 µM Incubation Duration: 48 hrs (hours) Experimental Results: Osteoblasts demonstrated the highest proliferation at 10 nM and 100 pM concentrations. Thirty-one nortriterpenoids, including 28 limonoids (1-28) and 3 degraded limonoids (29-31), and one diterpenoid (32), were isolated from the seed extract of Azadirachta indica (neem). Among these, six were new compounds and their structures were established to be 15-hydroxyazadiradione (3), 7-benzoyl-17-hydroxynimbocinol (5), 23-deoxyazadironolide (12), limocin E (13), 23-epilimocin E (14), and 7alpha-acetoxy-3-oxoisocopala-1,13-dien-15-oic acid (32). Upon evaluation of compounds 1-32 on the melanogenesis in the B16 melanoma cells, five compounds, 20, 26, 27, 29, and 31, exhibited marked inhibitory effect (74-91% reduction of melanin content at 25 microg/mL) with no or almost no toxicity to the cells. Seven compounds, 1, 6, 9, 10, 18, 20, and 26, on evaluation for their inhibitory effect against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 microg/ear) in mice, exhibited, except for compound 26, marked anti-inflammatory activity (ID(50) values 0.09-0.26 mg/ear). In addition, all of the 32 compounds exhibited moderate or potent inhibitory effects (IC(50) values of 230-501 mol ratio/32 pmol TPA) against the Epstein-Barr virus early antigen (EBV-EA) activation induced by TPA. Furthermore, on evaluation of azadirachtin B (21) for its anti-tumor-initiating activity on the two-stage carcinogenesis of mouse skin tumor induced by peroxynitrite (ONOO-; PN) as an initiator and TPA as a promoter, this exhibited marked inhibitory activity.[3] |
| Animal Protocol |
In vivo dose determination: Fifteen 1 to 2 day old pups were divided into five equal groups four equal groups and given a subcutaneous injection of either compound (0.5, 1.0, 5.0 and 10.0mg/kg/day) or equal volume of vehicle (normal saline) for 3 consecutive days. At the end of the treatment, pups were euthanized, and individual calvarias were harvested and cleaned of adherent tissue materials by gentle scrapping. Total RNA was isolated, and Q-PCR for ALP, RunX-2 and COL1 was performed as described earlier.[1]
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| References |
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| Additional Infomation |
Deacetylazadirachtinol has been reported in Azadirachta indica and Azadirachta excelsa with data available.
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| Molecular Formula |
C33H42O14
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|---|---|
| Molecular Weight |
662.685
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| Exact Mass |
662.257
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| Elemental Analysis |
C, 59.81; H, 6.39; O, 33.80
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| CAS # |
106500-25-8
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| PubChem CID |
21725521
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| Appearance |
White to off-white solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
780.5±60.0 °C at 760 mmHg
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| Flash Point |
246.6±26.4 °C
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| Vapour Pressure |
0.0±6.1 mmHg at 25°C
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| Index of Refraction |
1.628
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| LogP |
1.31
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
47
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| Complexity |
1490
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| Defined Atom Stereocenter Count |
16
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| SMILES |
C/C=C(\C)/C(=O)O[C@@H]1C[C@@H]([C@]23CO[C@@H]([C@H]2[C@]([C@@H]([C@H]4[C@H]3[C@@]1(CO4)C(=O)OC)O)(C)[C@@]56[C@@H]7C[C@H]([C@@]5(O6)C)[C@]8(C=CO[C@H]8O7)O)C(=O)OC)O
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| InChi Key |
USRBWQQLHKQWAV-ZGKQVQOISA-N
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| InChi Code |
InChI=1S/C33H42O14/c1-7-14(2)24(36)45-17-11-16(34)30-12-44-20(25(37)40-5)21(30)28(3,23(35)19-22(30)31(17,13-43-19)26(38)41-6)33-18-10-15(29(33,4)47-33)32(39)8-9-42-27(32)46-18/h7-9,15-23,27,34-35,39H,10-13H2,1-6H3/b14-7+/t15-,16+,17-,18+,19-,20+,21+,22-,23-,27+,28+,29+,30-,31+,32+,33+/m1/s1
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| Chemical Name |
dimethyl (2aR,2a1R,3S,4S,4aR,5S,7aS,8S,10R,10aS)-3,8-dihydroxy-4-((1aR,2S,3aS,6aS,7S,7aS)-6a-hydroxy-7a-methyl-3a,6a,7,7a-tetrahydro-2,7-methanofuro[2,3-b]oxireno[2,3-e]oxepin-1a(2H)-yl)-4-methyl-10-(((E)-2-methylbut-2-enoyl)oxy)octahydro-1H,7H-naphtho[1,8-bc
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| Synonyms |
Azadirachtin B; Azadirachtinol deacetyl; Azadirachtin-B; azadirachtin B; 106500-25-8; Deacetylazadirachtinol; AZADIRACHTIN B(SH); 3-Tigloylazadirachtol; Azadirachtinol deacetyl; UNII-X5T1RMZ28I; X5T1RMZ28I; Deacetylazadirachtinol
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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: (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)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~75.45 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.77 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (3.77 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5090 mL | 7.5450 mL | 15.0900 mL | |
| 5 mM | 0.3018 mL | 1.5090 mL | 3.0180 mL | |
| 10 mM | 0.1509 mL | 0.7545 mL | 1.5090 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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