AMXT-1501 tetra HCl is an orally bioactive polyamine transport inhibitor. AMXT-1501 blocks tumor growth in immunocompetent mice but not in athymic nude mice lacking T cells. DFMO and AMXT-1501 combined to induce caspase-3-mediated NB cell apoptosis.

Physicochemical Properties

Molecular Formula	C32H72CL4N6O2
Molecular Weight	714.77
CAS #	2444815-84-1
Appearance	Off-white to light yellow solid powder
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	Polyamine transport[1]
ln Vitro	The NB cell lines BE(2)-C, SMS-KCNR, and SH-SY5Y show cytotoxicity when treated with AMXT-1501 tetrahydrochloride (0.39-50 µM; 48 hours); the IC50 values for SMS-KCNR are 17.72 µM, BE(2)-C is 17.69 µM, and SH-SY5Y is 14.13 µM[2]. The AMXT-1501 tetrahydrochloride (2.5 µM) and DFMO (2.5 mM) are applied to BE(2)-C, SMS-KCNR, and SH-SY5Y cells either separately or in combination (AMXT-1501 tetrahydrochloride 2.5 µM + DFMO 2.5 mM). The levels of cleaved PARP, cleaved caspase 3, and noncleaved PARP do not change significantly after 96 hours of exposure to either AMXT-1501 tetrahydrochloride or DFMO. However, cells treated with the combination of AMXT-1501 tetrahydrochloride and DFMO show a decrease in noncleaved PARP and an increase in cleaved PARP and cleaved caspase 3[2].
ln Vivo	AMXT-1501 tetrahydrochloride (3 mg/kg; subcutaneous injection; daily; 28 days) by itself is adequate to moderately postpone the onset of EAE, but it is unable to prevent animals from reaching the endpoint. Nonetheless, the tetrahydrochloride AMXT-1501 and DFMO alone are adequate to reduce the polyamine pool in T cells, which in turn inhibits T cell proliferation and effector function in vivo[3].
Cell Assay	Cell Viability Assay[2] Cell Types: BE(2)-C, SMS‐KCNR and SH‐SY5Y cells Tested Concentrations: 0.39 µM, 1 µM, 3.1 µM, 10 µM, 31 µM, 50 µM Incubation Duration: 48 hrs (hours) Experimental Results: AMXT-1501 tetrahydrochloride demonstrated cytotoxicity against this panel of NB cell lines. Western Blot Analysis[2] Cell Types: BE(2)‐C, SMS‐KCNR and SH‐SY5Y cells Tested Concentrations: 2.5 µM Incubation Duration: 72 hrs (hours) Experimental Results: Combination treatment with DFMO diminished the amount of noncleaved PARP and increased the amount of cleaved PARP and cleaved caspase 3 in all three cell lines.
Animal Protocol	Animal/Disease Models: C57BL/6 (WT) and ODC knockout strain (ODC cKO) mice bearing experimental autoimmune encephalomyelitis (EAE) model [ 3] Doses: 3 mg/kg Route of Administration: subcutaneous (sc) injection; every day; 28 days Experimental Results: Displayed a delayed disease onset initially, but eventually proceeded with pathologic development and reached the endpoint.
References	[1]. Polyamine-blocking therapy reverses immunosuppression in the tumor microenvironment. Cancer Immunol Res. 2014 Mar;2(3):274-85. [2]. AMXT‐1501, a novel polyamine transport inhibitor, synergizes with DFMO in inhibiting neuroblastoma cell proliferation by targeting both ornithine decarboxylase and polyamine transport. Int J Cancer. 2013 Sep 15;133(6):1323-33. [3]. De novo synthesis and salvage pathway coordinately regulate polyamine homeostasis and determine T cell proliferation and function. Sci Adv. 2020 Dec 16;6(51):eabc4275.

Solubility Data

Solubility (In Vitro)	H2O :~83.33 mg/mL (~116.58 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 50 mg/mL (69.95 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.3991 mL	6.9953 mL	13.9905 mL
	5 mM	0.2798 mL	1.3991 mL	2.7981 mL
	10 mM	0.1399 mL	0.6995 mL	1.3991 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.