Boc-Leu-Gly-Arg-AMC is a fluorescent AMC substrate for invertase and may be utilized in enzymatic assays.

Physicochemical Properties

Molecular Formula	C29H43N7O7
Molecular Weight	601.6944
Exact Mass	601.322
CAS #	65147-09-3
PubChem CID	53765602
Appearance	White to off-white solid powder
LogP	4.28
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	15
Heavy Atom Count	43
Complexity	1080
Defined Atom Stereocenter Count	2
SMILES	CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)OC(C)(C)C
InChi Key	DLAQYOHJZUTWDJ-SFTDATJTSA-N
InChi Code	InChI=1S/C29H43N7O7/c1-16(2)12-21(36-28(41)43-29(4,5)6)25(39)33-15-23(37)35-20(8-7-11-32-27(30)31)26(40)34-18-9-10-19-17(3)13-24(38)42-22(19)14-18/h9-10,13-14,16,20-21H,7-8,11-12,15H2,1-6H3,(H,33,39)(H,34,40)(H,35,37)(H,36,41)(H4,30,31,32)/t20-,21-/m0/s1
Chemical Name	tert-butyl N-[(2S)-1-[[2-[[(2S)-5-(diaminomethylideneamino)-1-[(4-methyl-2-oxochromen-7-yl)amino]-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-4-methyl-1-oxopentan-2-yl]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 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

ln Vitro

In order to prove that the abdominal glands contain proteolytic enzymes that can produce Sodefrin, an enzymatic test was created with the synthetic substrate Boc-Leu-Gly-Arg-AMC. The hydrolysis of Boc-Leu-Gly-Arg-AMC by crude extracts of the abdominal glands releases 7-amino-4-methylcoumarin, a sign that the gland contains an enzyme that converts sodefrin from its precursor molecule. At pH 9.0 and 45°C, the extract's ability to cleave Boc-Leu-Gly-Arg-AMC is at its peak [1].

References

[1]. Evidence for processing enzymes in the abdominal gland of the newt, Cynops pyrrhogaster, that generate sodefrin from its biosynthetic precursor. Zoolog Sci. 2007 May;24(5):521-4. [2]. Proteolytic enzymes in coastal surface seawater: Significant activity of endopeptidases and exopeptidases Limnol. Oceanogr., 50(2), 2005, 722-726.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~166.20 mM) H2O : < 0.1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.15 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.15 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.15 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6620 mL	8.3099 mL	16.6199 mL
	5 mM	0.3324 mL	1.6620 mL	3.3240 mL
	10 mM	0.1662 mL	0.8310 mL	1.6620 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.