Cycleanine is a potent vasculoselective Ca-antagonist. Cycleanine has pain-relieving, muscle-relaxing, and anti-inflammatory activities. Cycleanine may be used to exert anti-ovarian cancer effects through the apoptotic pathway.

Physicochemical Properties

Molecular Formula	C38H42N2O6
Molecular Weight	622.75
Exact Mass	622.304
CAS #	518-94-5
PubChem CID	121313
Appearance	White to off-white solid powder
Density	1.2±0.1 g/cm3
Boiling Point	691.6±55.0 °C at 760 mmHg
Flash Point	169.8±28.7 °C
Vapour Pressure	0.0±2.2 mmHg at 25°C
Index of Refraction	1.586
LogP	3.96
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	4
Heavy Atom Count	46
Complexity	895
Defined Atom Stereocenter Count	2
SMILES	CN1CCC2=CC(=C(C3=C2[C@H]1CC4=CC=C(C=C4)OC5=C6[C@@H](CC7=CC=C(O3)C=C7)N(CCC6=CC(=C5OC)OC)C)OC)OC
InChi Key	ANOXEUSGZWSCQL-LOYHVIPDSA-N
InChi Code	InChI=1S/C38H42N2O6/c1-39-17-15-25-21-31(41-3)35(43-5)37-33(25)29(39)19-23-7-11-28(12-8-23)46-38-34-26(22-32(42-4)36(38)44-6)16-18-40(2)30(34)20-24-9-13-27(45-37)14-10-24/h7-14,21-22,29-30H,15-20H2,1-6H3/t29-,30-/m1/s1
Chemical Name	(11R,26R)-4,5,19,20-tetramethoxy-10,25-dimethyl-2,17-dioxa-10,25-diazaheptacyclo[26.2.2.213,16.13,7.118,22.011,36.026,33]hexatriaconta-1(31),3(36),4,6,13,15,18(33),19,21,28(32),29,34-dodecaene
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 (e.g. under nitrogen), avoid exposure to moisture and light.
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	L-type calcium channel
ln Vitro	In a voltage- and frequency-dependent way, cycleanine suppresses the L-type Ca-current (ICaL) of single rat ventricular cardiomyocytes[1]. Cycleanine is slightly less effective than cancer cells against normal human OSE cells[2]. For Ovcar-8, A2780, Igrov-1, and Ovcar-4 cell lines, (20 μM; 48 hours) demonstrates cytotoxicity with IC50s ranging from 7 to 14 μM[2]. Significant PARP cleavage, a hallmark of apoptosis, is caused by cycleanine (20 μM; 24 hours)[2]. Cycleanine (20 μM; 48 hours) significantly increases the population of cells that undergo early and late apoptosis[2]. Cytotoxicity Assay for Cycleanine Cells [2] Cell line: Igrov-1, A2780, Ovcar-8, and Ovcar-4 cells. Focus: 20 micrograms 48-hour incubation period Result shown cytotoxicity against Ovcar-8, A2780, Igrov-1, and Ovcar-4 cell lines, with IC50 values of 10 μM, 7.6 μM, 14 μM, and 7.2 μM, in that order.
ln Vivo	At an IC50 of 0.8 nM, cycleanine prevents rabbit aortic ring contraction caused by KCl[1].
Cell Assay	Western Blot Analysis[2] Cell Types: Ovcar-8 cells Tested Concentrations: 20 μM Incubation Duration: 24 hrs (hours) Experimental Results: Induced 1.1-fold increase in PARP-1 cleavage compared with carboplatin. Apoptosis Analysis[2] Cell Types: Ovcar-8 cells Tested Concentrations: 20 μM Incubation Duration: 48 hrs (hours) Experimental Results: Caused a significant increase of the population of both early and late apoptotic cells. Cell Cycle Analysis[2] Cell Types: Ovcar-8 cells Tested Concentrations: 20 μM Incubation Duration: 48 hrs (hours) Experimental Results: Increased the percentage of Ovcar-8 cells in subG1.
References	[1]. Calcium antagonist properties of the bisbenzylisoquinoline alkaloid cycleanine. Fundam Clin Pharmacol. 1998;12(2):182-7. [2]. Cytotoxicity Effects and Apoptosis Induction by Bisbenzylisoquinoline Alkaloids from Triclisia subcordata. Phytother Res. 2016 Sep;30(9):1533-9.
Additional Infomation	Cycleanine is a member of isoquinolines and a bisbenzylisoquinoline alkaloid. Cycleanine has been reported in Stephania tetrandra, Cyclea tonkinensis, and other organisms with data available.

Solubility Data

Solubility (In Vitro)

DMSO : 5 mg/mL (8.03 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	1.6058 mL	8.0289 mL	16.0578 mL
	5 mM	0.3212 mL	1.6058 mL	3.2116 mL
	10 mM	0.1606 mL	0.8029 mL	1.6058 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.