BW-180C ([D-Ala2, D-Leu5]-Enkephalin) is a water-soluble (H2O-soluble) peptide opioid receptor (Opioid Receptor) agonist.

Physicochemical Properties

Molecular Formula	C29H39N5O7
Molecular Weight	569.65
Exact Mass	569.285
CAS #	63631-40-3
PubChem CID	6917707
Appearance	White to off-white solid powder
LogP	2.489
Hydrogen Bond Donor Count	7
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	15
Heavy Atom Count	41
Complexity	885
Defined Atom Stereocenter Count	4
SMILES	C[C@H](C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N
InChi Key	ZHUJMSMQIPIPTF-IBURTVSXSA-N
InChi Code	InChI=1S/C29H39N5O7/c1-17(2)13-24(29(40)41)34-28(39)23(15-19-7-5-4-6-8-19)33-25(36)16-31-26(37)18(3)32-27(38)22(30)14-20-9-11-21(35)12-10-20/h4-12,17-18,22-24,35H,13-16,30H2,1-3H3,(H,31,37)(H,32,38)(H,33,36)(H,34,39)(H,40,41)/t18-,22+,23+,24-/m1/s1
Chemical Name	(2R)-2-[[(2S)-2-[[2-[[(2R)-2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]propanoyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoic 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: Please store this product in a sealed and protected environment, 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)

Biological Activity

Targets	Opioid Receptor[1]
ln Vitro	In SH-SY5Y cells, BW-180C ([D-Ala2, D-Leu5]-Enkephalin) dramatically decreases cellular transcription without endangering the cells. After primary neurons recover for 72 hours in the absence of BW-180C, transcriptional activity fully resumes[1].
ln Vivo	BW-180C ([D-Ala2, D-Leu5]-Enkephalin) defends against IR damage in hepatocytes but not in rat liver sinusoidal endothelial cells. Although the BW-180C group's GPT levels are much lower than the Control group's, the two groups' serum HA levels are the same. In comparison to the Control group, the BW-180C group's liver tissue MDA concentrations are noticeably lower[2]. The activation of both mu- and delta-opioid receptors mediates the analgesia brought on by BW-180C (DADLE)[3].
Cell Assay	Cell Viability Assay[1] Cell Types: Human neuroblastoma cells, SH-SY5Y cells Tested Concentrations: 100 pM to 10 μM Incubation Duration: 24-72 hrs (hours) Experimental Results: Dramatically inhibited cellular transcription. Western Blot Analysis[1] Cell Types: Primary Cortical neurons Tested Concentrations: 100 nM Incubation Duration: 72 hrs (hours) Experimental Results: Treatment Dramatically diminished phosphorylation of RNA polymerase II at both Ser 2 and Ser 5.
Animal Protocol	Animal/Disease Models: Male Wistar rats weighing 200 to 250 g[2] Doses: 5 mg/kg Route of Administration: Administered intravenously (iv) into the inferior vena cava Experimental Results: After 120 min of reperfusion, the serum GPT levels were Dramatically lower in the DADLE group than in the Control group. Animal/Disease Models: Male C57BL/6 mice (age, 35-42 days; weight, 18-22 g; 6 mice in each group)[3] Doses: 5 mg/kg Route of Administration: Injected intraperitoneally (ip) 15 min prior to I/R Experimental Results: The serum levels of AST and ALT in the serum DADLE group were Dramatically lower.
References	[1]. [D-Ala2, D-Leu5] encephalin (DADLE) reversibly inhibits cellular transcription in neurons without causing cell injury. Brain Res. 2014 May 27;1565:1-7. [2]. [D-Ala2, D-Leu5] enkephalin (DADLE) protects liver against ischemia-reperfusion injury in the rat. J Surg Res. 2003 Sep;114(1):72-7. [3]. DADLE improves hepatic ischemia/reperfusion injury in mice via activation of the Nrf2/HO?1 pathway. Mol Med Rep. 2017 Nov;16(5):6214-6221.
Additional Infomation	A delta-selective opioid (ANALGESICS, OPIOID). It can cause transient depression of mean arterial blood pressure and heart rate. A delta-selective opioid (ANALGESICS, OPIOID). It can cause transient depression of mean arterial blood pressure and heart rate.

Solubility Data

Solubility (In Vitro)

DMSO: 250 mg/mL (438.87 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	1.7555 mL	8.7773 mL	17.5546 mL
	5 mM	0.3511 mL	1.7555 mL	3.5109 mL
	10 mM	0.1755 mL	0.8777 mL	1.7555 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.