(rel)-Asperparaline A ((rel)-Aspergillimide) is an anthelmintic metabolite extracted from okara fermented by Aspergillus japonicas JV-23. (rel)-Asperparaline A is also a potent and specific nAChR antagonist. (rel)-Asperparaline A displays paralyzing effects in silkworms.

Physicochemical Properties

Molecular Formula	C20H29N3O3
Molecular Weight	359.46
Exact Mass	359.22
CAS #	195966-93-9
PubChem CID	154701594
Appearance	Typically exists as solid at room temperature
Density	1.3±0.1 g/cm3
Boiling Point	546.1±50.0 °C at 760 mmHg
Flash Point	244.6±22.5 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.615
LogP	1.11
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	0
Heavy Atom Count	26
Complexity	763
Defined Atom Stereocenter Count	4
SMILES	O=C1C23[C@@]([H])(C([H])([H])[H])C([H])([H])C([H])([H])N2C([H])([H])[C@@]2(C([H])([H])[C@@]4(C(N(C([H])([H])[H])C(C4([H])[H])=O)=O)C(C([H])([H])[H])(C([H])([H])[H])[C@]2([H])C3([H])[H])N1C([H])([H])[H]
InChi Key	RTNMRJRMTGSUAE-UXVBPGSQSA-N
InChi Code	InChI=1S/C20H29N3O3/c1-12-6-7-23-11-19-10-18(9-14(24)21(4)15(18)25)17(2,3)13(19)8-20(12,23)16(26)22(19)5/h12-13H,6-11H2,1-5H3/t12-,13-,18-,19+,20?/m0/s1
Chemical Name	(1S,6S,9S,11S)-1',6,10,10,13-pentamethylspiro[3,13-diazatetracyclo[5.5.2.01,9.03,7]tetradecane-11,3'-pyrrolidine]-2',5',14-trione
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	In the neurons of silkworm larvae, asperparaline A (1 μM; 1 min) significantly and reversibly inhibits the current generated by acetylcholine (Ach; 10 µM)[2]. With IC50s of 20.2 and 39.6 nM, respectively, asperparaline A (1-1000 nM; 1 min) differentially blocks the peak and gradually desensitizing currents[2]. Asperparaline A (10 μM; 1 min) marginally affects the amplitudes of the responses to ACh of the α4β2 and α7 nAChRs in X, but decreases the peak current amplitude of the ACh (100 µM)-induced response of α3β4 nAChR by 33.4%. Ovum laevis [2].
ln Vivo	Asperparaline A (10 μg/g; po) acts against silk worms to cause paralysis after one hour and continues to do so for seven to ten hours[1]. Asperparaline A (3 μg/g; injection with a micro-syringe) paralyzes silk worms within 20 minutes and keeps the effect going for 4 to 5 hours[1].
References	[1]. New paralytic alkaloids, asperparalines A, B and C, from Aspergillus japonicus JV-23. Biosci Biotechnol Biochem. 2000 Jan;64(1):111-5. [2]. A fungal metabolite asperparaline a strongly and selectively blocks insect nicotinic acetylcholine receptors: the first report on the mode of action. PLoS One. 2011 Apr 1;6(4):e18354.

Solubility Data

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	2.7820 mL	13.9098 mL	27.8195 mL
	5 mM	0.5564 mL	2.7820 mL	5.5639 mL
	10 mM	0.2782 mL	1.3910 mL	2.7820 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.