Voreloxin (SNS-595) HCl 175519-16-1_Topoisomerase_DNA Damage_Signaling Pathways_Products

Voreloxin HCl (formerly known as AG-7352; SNS-595; SPC 595; AG 7352; SNS595, AG-7352; Vosaroxin), the hydrochloride salt of Voreloxin which is a naphthyridine analog, is a novel and potent Topoisomerase II inhibitor with a broad-spectrum antineoplastic activity. Vosaroxin is an experimental medication that has been given orphan status to treat ovarian cancer and acute myelogenous leukemia (AML). It works by selectively intercalating into DNA to obstruct topoisomerase II's re-ligation process during DNA replication. The result is the inhibition of RNA, protein, and DNA replication, which is followed by cell cycle arrest at the G2 phase and p53-independent apoptosis.

Physicochemical Properties

Molecular Formula

C18H20CLN5O4S

Molecular Weight

437.9

Exact Mass

437.092

CAS #

175519-16-1

Related CAS #

175414-77-4

PubChem CID

10343042

Appearance

White to gray solid powder

LogP

2.221

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

662

Defined Atom Stereocenter Count

SMILES

CN[C@H]1CN(C[C@@H]1OC)C2=NC3=C(C=C2)C(=O)C(=CN3C4=NC=CS4)C(=O)O.Cl

InChi Key

JJZCCQHWCOXGCL-QNTKWALQSA-N

InChi Code

InChI=1S/C18H19N5O4S.ClH/c1-19-12-8-22(9-13(12)27-2)14-4-3-10-15(24)11(17(25)26)7-23(16(10)21-14)18-20-5-6-28-18;/h3-7,12-13,19H,8-9H2,1-2H3,(H,25,26);1H/t12-,13-;/m0./s1

Chemical Name

7-[(3S,4S)-3-methoxy-4-(methylamino)pyrrolidin-1-yl]-4-oxo-1-(1,3-thiazol-2-yl)-1,8-naphthyridine-3-carboxylic acid;hydrochloride

Synonyms

Vosaroxin; SNS 595; SPC595; AG7352; SNS595; SPC-595; AG-7352; SNS-595; SPC 595; AG 7352; VVosaroxin

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, avoid exposure to moisture.

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

Topoisomerase II

ln Vitro

Voreloxin Hydrochloride is a novel topoisomerase II poison and inhibitor that causes site-specific DNA DSB, G2 arrest, and apoptosis by intercalating DNA. Voreloxin (0.1–20 µM) causes site-selective DNA DSB in CCRF-CEM cells and inhibits topoisomerase II activity. Voreloxin (0.11, 0.33, 1, 3 µM) partially induces G2 arrest in A549 lung cancer cell line by using topoisomerase II. Voreloxin cytotoxic activity requires DNA intercalation. Nevertheless, Voreloxin (1–9 µM) does not produce a noticeable amount of ROS[1]. In AML cell lines MV4-11 and HL-60, voreloxin exhibits strong cytotoxic activity, with IC50 values of 95 ± 8 nM and 884 ± 114 nM, respectively. In acute leukemia cell lines, voreloxin and cytarabine exhibit additive or synergistic activity[2]. With a mean LD50 of 2.3 μM, voreloxin exhibits activity against primary acute myeloid leukemia (AML). In the myeloid cell lines NB4 and HL-60, voreloxin has an LD50 of 0.59 μM ± 0.25 μM. Voreloxin acts on topoisomerase II and induces cell accumulation in the S and G2 phases of the cell cycle[3].

ln Vivo

Voreloxin (20 mg/kg, i.v.) administered once every four days, twice (q4d ×2), is the only treatment that reduces the cellularity of CD-1 mice's bone marrow by 80%. In mice, voreloxin at 10 mg/kg combined with cytarabine results in marrow ablation, sinusoidal dilatation, and adipocyte infiltration. In bone marrow and peripheral blood CD-1 mice, voreloxin (20 mg/kg, i.v.) and cytarabine induce a reversible decrease in myeloid and lymphoid cells. When voreloxin (10 mg/kg, q4d ×2) and cytarabine are combined, platelets CD-1 mice experience a more mild reversible neutropenia[2].

Cell Assay

MTS cell proliferation assays are used to conduct in vitro toxicity tests on primary AML mononuclear cells over the course of 48 hours. One can calculate lethal doses (LD50). In a final volume of 90 μL, cells are treated with voreloxin (31.25 nM to 4 μM) and Ara-C (62.5 nM to 8 μM) by serial dilution and incubated for 48 hours. After incubation, the reaction is incubated for an additional 4 hours with the addition of 20 μL of MTS reagent. Spectrophotometry is used to measure the absorbance of the reaction at 490 nm after this point, and the percentage of viable cells is computed in relation to the untreated control cells in the same experiment. Software called Calcusyn is used to calculate IC50 values[3].

Animal Protocol

Before beginning therapy, the animals are weighed, randomized according to their body weight, and placed into study groups. On day zero and day four (q4d ×2), voreloxin is given intravenously (IV) at a dose of 10 or 20 mg/kg each time. Day zero and day four (tid q4d ×2) are given cytarabine subcutaneously (SC) at 20 or 60 mg/kg every 8 hours. Three or fewer animals per treatment group, but no more than ten, have their tissues and blood sampled on days six, eight, and twelve. Femurs are immersed for 24 to 48 hours in either the Streck Tissue Fixative solution or the 10% formalin solution, and then they are dehydrated to 70% (ethanol, isopropanol, or methanol). At Biopathology Labs, femurs undergo decalcification, paraffin embedding, and sectioning. Hematoxylin-eosin is used to stain the four micron sections (H&E). Femurs stained with H&E are inspected, and the bone marrow's percentage cellularity is calculated. Using Image-Pro Plus v6.1 software, digital photos of representative femur sections are captured on a Leica DM2000 microscope[2].

References

[1]. Vosaroxin is a novel topoisomerase-II inhibitor with efficacy in relapsed and refractory acute myeloid leukaemia. Expert Opin Pharmacother. 2015 Jun;16(9):1395-402.

[2]. Voreloxin, a first-in-class anticancer quinolone derivative, acts synergistically with cytarabine in vitro and induces bone marrow aplasia in vivo. Cancer Chemother Pharmacol. 2010 Oct;66(5):881-8.

[3]. The topoisomerase II inhibitor voreloxin causes cell cycle arrest and apoptosis in myeloid leukemia cells and acts in synergy with cytarabine. Haematologica. 2011 Mar;96(3):393-9.

Solubility Data

Solubility (In Vitro)

DMSO: ~1 mg/mL (~2.3 mM)

Water: ~1 mg/mL (~2.3 mM)

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2 mg/mL (4.57 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2836 mL	11.4181 mL	22.8363 mL
	5 mM	0.4567 mL	2.2836 mL	4.5673 mL
	10 mM	0.2284 mL	1.1418 mL	2.2836 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.

PeptideDB

Voreloxin (SNS-595) HCl 175519-16-1

CAS No.: 175519-16-1

Physicochemical Properties

Biological Activity

Solubility Data