Epibrassinolide (24-Epibrassinolide; B1105; BP55) is a naturally occurring plant growth hormone which shows great potential to alleviate heavy metals and pesticide stress in plants. Epibrassinolide has the potential to induce apoptosis in a variety of cancer cells without impairing the growth of normal cells.

Physicochemical Properties

Molecular Formula	C₂₈H₄₈O₆
Molecular Weight	480.68
Exact Mass	480.345
CAS #	78821-43-9
Related CAS #	Brassinolide;72962-43-7
PubChem CID	443055
Appearance	White to off-white solid
Density	1.1±0.1 g/cm3
Boiling Point	633.7±55.0 °C at 760 mmHg
Melting Point	256°C
Flash Point	202.3±25.0 °C
Vapour Pressure	0.0±4.2 mmHg at 25°C
Index of Refraction	1.536
LogP	3.12
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	34
Complexity	755
Defined Atom Stereocenter Count	13
SMILES	C[C@@]12[C@@H]([C@H](C)[C@@H](O)[C@H](O)[C@H](C)C(C)C)CC[C@H]1[C@@H]1COC(=O)[C@H]3C[C@@H]([C@@H](C[C@]3(C)[C@H]1CC2)O)O
InChi Key	IXVMHGVQKLDRKH-QHBHMFGVSA-N
InChi Code	InChI=1S/C28H48O6/c1-14(2)15(3)24(31)25(32)16(4)18-7-8-19-17-13-34-26(33)21-11-22(29)23(30)12-28(21,6)20(17)9-10-27(18,19)5/h14-25,29-32H,7-13H2,1-6H3/t15-,16+,17+,18-,19+,20+,21-,22+,23-,24-,25-,27-,28-/m1/s1
Chemical Name	(1S,2R,4R,5S,7S,11S,12S,15R,16S)-15-[(2S,3R,4R,5R)-3,4-dihydroxy-5,6-dimethylheptan-2-yl]-4,5-dihydroxy-2,16-dimethyl-9-oxatetracyclo[9.7.0.02,7.012,16]octadecan-8-one
Synonyms	24-Epibrassinolide; B1105; BP55;B 1105; BP 55; B-1105; BP-55
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)

Biological Activity

Targets	Plant growth regulator; Apoptosis inducer
ln Vitro	Epibrassinolide (EBR) is a bioactive molecule from the brassinosteroid family of plant growth regulators. Cells were treated with epibrassinolide at varying concentrations (0-100 μM) for 24 or 48 hours, and cell viability was assessed by MTT. In contrast to untreated samples in LNCaP and DU145 gun molds, epibrassinosteroid caused cell viability death in a dose and time-dependent manner. Increasing epibrassinolide concentrations caused higher cell viability loss in LNCaP cells than DU145 cells, showing that epibrassinolide has a greater influence on androgenic cell proliferation and cell death than androgenic products. Further trials used 25 μM epibrassinolide, which produced the most cytotoxic effects on both cell types. Calculate cell data for 96 hours to see how Epibrassinolide therapy affects cell proliferation. LNCaP cells showed a higher and earlier suppression of cell growth compared to DU145 cells [1].
Cell Assay	The PNT1a normal prostate epithelial cell line is utilized. Cells are grown in RPMI 1640 medium supplemented with 10% fetal bovine serum, 10,000 U penicillin/mL, and 10 mg streptomycin/mL. (HERA cell 150) Cells are cultured at 37°C in an incubator with humidified 5% CO2. The desired drug concentrations are applied after cells have been seeded overnight. In 96-well plates, cells are seeded at a density of 1×104 per well and exposed to various Epibrassinolide concentrations for 24 and 48 hours. Each well receives 10 μL of MTT dye (5 mg/mL), and cells are incubated at 37°C for 4 hours. In 200 L DMSO, the formed formazan crystals are solubilized. At 570 nm, spectrophotometric measurements of the solubilized formazan's density are made[1].
References	[1]. Role of 24-epibrassinolide (EBL) in mediating heavy metal and pesticide induced oxidative stress in plants: A review. Ecotoxicol Environ Saf. 2018 Jan;147:935-944. [2]. Activation of polyamine catabolic enzymes involved in diverse responses against epibrassinolide-induced apoptosis in LNCaP and DU145 prostate cancer cell lines.Amino Acids. 2014 Mar;46(3):553-64.
Additional Infomation	24-epi-brassinolide is a 2alpha-hydroxy steroid, a 3alpha-hydroxy steroid, a 22-hydroxy steroid, a 23-hydroxy steroid and a brassinosteroid. 24-epi-Brassinolide has been reported in Vicia faba, Gypsophila perfoliata, and other organisms with data available. See also: Brassinolide (annotation moved to).

Solubility Data

Solubility (In Vitro)

DMSO: 96~100 mg/mL (199.7~208.0 mM) Ethanol: ~16 mg/mL (~33.3 mM)

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.0804 mL	10.4019 mL	20.8039 mL
	5 mM	0.4161 mL	2.0804 mL	4.1608 mL
	10 mM	0.2080 mL	1.0402 mL	2.0804 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.