Shishijimicin A is a new anti-tumor agent discovered from the ascomycete Didemnum proliferum. Shishijimicin A displays anti-tumor effects and is highly cytotoxic. Shishijimicin A is a click chemical reagent. It has Alkyne groups and could undergo CuAAc (copper-catalyzed azide-alkyne cycloaddition reaction) with compounds bearing Azide groups.

Physicochemical Properties

Molecular Formula	C46H52N4O12S4
Molecular Weight	981.184687614441
Exact Mass	980.246
CAS #	503860-50-2
PubChem CID	21577974
Appearance	Typically exists as solid at room temperature
LogP	3.7
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	18
Rotatable Bond Count	16
Heavy Atom Count	66
Complexity	2050
Defined Atom Stereocenter Count	10
SMILES	COC1C(NC(C)C)COC(OC2C(O)C(C(C3C4=C(C5C(N4)=CC=C(O)C=5)C=CN=3)=O)(SC)C(C)OC2OC2C3C(C(CC(C=3NC(OC)=O)=O)(O)C#CC=CC#C2)=CCSSSC)C1
InChi Key	FVNYJZKDBPDHIP-NATOZQEOSA-N
InChi Code	InChI=1S/C46H52N4O12S4/c1-24(2)48-31-23-59-35(21-34(31)57-4)62-40-42(54)46(63-6,41(53)39-37-27(15-18-47-39)28-20-26(51)13-14-30(28)49-37)25(3)60-43(40)61-33-12-10-8-9-11-17-45(56)22-32(52)38(50-44(55)58-5)36(33)29(45)16-19-65-66-64-7/h8-9,13-16,18,20,24-25,31,33-35,40,42-43,48-49,51,54,56H,19,21-23H2,1-7H3,(H,50,55)/b9-8-,29-16+/t25-,31+,33+,34+,35+,40-,42-,43+,45+,46-/m1/s1
Chemical Name	methyl N-[(1R,4Z,8S,13E)-1-hydroxy-8-[(2R,3R,4R,5R,6R)-4-hydroxy-5-(6-hydroxy-9H-pyrido[3,4-b]indole-1-carbonyl)-3-[(2S,4S,5S)-4-methoxy-5-(propan-2-ylamino)oxan-2-yl]oxy-6-methyl-5-methylsulfanyloxan-2-yl]oxy-13-[2-(methyltrisulfanyl)ethylidene]-11-oxo-10-bicyclo[7.3.1]trideca-4,9-dien-2,6-diynyl]carbamate
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

ln Vitro	High cytotoxicity is demonstrated by shishijimicin A (compound 1) (0.47-2.0 pg/mL) against 3Y1, Hela, and P388 cells [1]. High cytotoxicity is demonstrated by shishijimicin A (0.13-1000 nM) against MES SA, MES SA DX, and HEK 293T cells [2].
Cell Assay	Cytotoxicity assay [1] Cell Types: 3Y1, Hela and P388 Cell Tested Concentrations: 0.47-2.0 pg/mL Incubation Duration: Experimental Results: The IC50 values of 3Y1, Hela and P388 cells were 2.0, 1.8 and 0.47 pg/mL respectively. Cytotoxicity assay[2] Cell Types: MES SA, MES SA DX and HEK 293T Cell Tested Concentrations: 0.13-1000 nM Incubation Duration: Experimental Results: IC50 values for MES SA, MES SA DX and HEK were 0.13, >1000 and 0.016 nM respectively for HEK 293T cells.
References	[1]. Shishijimicins A-C, novel enediyne antitumor antibiotics from the ascidian Didemnum proliferum(1). J Am Chem Soc. 2003 Feb 26;125(8):2044-5. [2]. Streamlined Total Synthesis of Shishijimicin A and Its Application to the Design, Synthesis, and Biological Evaluation of Analogues thereof and Practical Syntheses of PhthNSSMe and Related Sulfenylating Reagents. J Am Chem Soc. 2018 Sep 26;140(38):12120-12136.
Additional Infomation	methyl N-[(1R,4Z,8S,13E)-1-hydroxy-8-[(2R,3R,4R,5R,6R)-4-hydroxy-5-(6-hydroxy-9H-pyrido[3,4-b]indole-1-carbonyl)-3-[(2S,4S,5S)-4-methoxy-5-(propan-2-ylamino)oxan-2-yl]oxy-6-methyl-5-methylsulfanyloxan-2-yl]oxy-13-[2-(methyltrisulfanyl)ethylidene]-11-oxo-10-bicyclo[7.3.1]trideca-4,9-dien-2,6-diynyl]carbamate has been reported in Didemnum fragile with data available.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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	1.0192 mL	5.0959 mL	10.1918 mL
	5 mM	0.2038 mL	1.0192 mL	2.0384 mL
	10 mM	0.1019 mL	0.5096 mL	1.0192 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.