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Evofosfamide (TH-302)

Cat. No.: S2757

Evofosfamide (TH-302) is a selective hypoxia-activated prodrug targeting hypoxic regions of solid tumors with IC50 of 19 nM. It demonstrates 270-fold enhanced cytotoxicity under hypoxia versus their potency under aerobic conditions, and is stable to cytochrome P450 metabolism.
Evofosfamide (TH-302) Chemical Structure

Estructura química

Peso molecular: 449.04

Saltar a

Control de calidad (Quality Control)

Lote: Pureza: 99.82%
99.82

Cultivo celular, tratamiento y concentración de trabajo
(Cell Culture, Treatment & Working Concentration)

Líneas celulares Tipo de ensayo Concentración Tiempo de incubación Formulación Descripción de la actividad PMID
H460 Cytotoxicity assay 2 hrs Cytotoxicity against human H460 cells under hypoxic condition after 2 hrs by Alamar blue staining assay, IC50=0.019μM. 18257544
H460 Cytotoxicity assay 2 hrs Cytotoxicity against human H460 cells under hypoxic condition after 2 hrs by clonogenic assay, IC90=0.1μM. 18257544
HT29 Cytotoxicity assay 2 hrs Cytotoxicity against human HT29 cells under hypoxic condition after 2 hrs by clonogenic assay, IC90=0.2μM. 18257544
H460 Cytotoxicity assay 2 hrs Cytotoxicity against human H460 cells under normoxic condition after 2 hrs by Alamar blue staining assay, IC50=5.1μM. 18257544
H460 Cytotoxicity assay 2 hrs Cytotoxicity against human H460 cells under normoxic condition after 2 hrs by clonogenic assay, IC90=30μM. 18257544
HT29 Cytotoxicity assay 2 hrs Cytotoxicity against human HT29 cells under normoxic condition after 2 hrs by clonogenic assay, IC90=40μM. 18257544
NCI-H460 Cytotoxicity assay 24 hrs Cytotoxicity against human NCI-H460 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay, IC50=9.08μM. 28350997
HT-29 Cytotoxicity assay 24 hrs Cytotoxicity against human HT-29 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay, IC50=49.51μM. 28350997
NCI-H460 Cytotoxicity assay 24 hrs Cytotoxicity against human NCI-H460 cells after 24 hrs under hypoxic condition by luminescence-based Cell-Titer Glo assay, IC50=0.0093μM. 29079474
NCI-H460 Cytotoxicity assay 24 hrs Cytotoxicity against human NCI-H460 cells after 24 hrs under normoxic condition by luminescence-based Cell-Titer Glo assay, IC50=6.65μM. 29079474
DU145 Cytotoxicity assay 2 hrs Cytotoxicity against human DU145 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay, IC50=4.14μM. 29259746
PC3 Cytotoxicity assay 2 hrs Cytotoxicity against human PC3 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay, IC50=6.49μM. 29259746
HEMC-SS Antiproliferative assay 24 hrs Antiproliferative activity against human HEMC-SS cells treated for 24 hrs under hypoxic condition followed by compound wash-out and incubated under normoxic condition for 48 hrs by alamar blue assay, IC50=0.13μM. 30199705
HEMC-SS Antiproliferative assay 24 hrs Antiproliferative activity against human HEMC-SS cells treated for 24 hrs under normoxic condition followed by compound wash-out and incubated under normoxic condition for 48 hrs by alamar blue assay, IC50=2.9μM. 30199705
NCI-H460 Antiproliferative assay Antiproliferative activity against human NCI-H460 cells in presence of N2, IC50=2μM. 30295477
NCI-H460 Antiproliferative assay Antiproliferative activity against human NCI-H460 cells in presence of O2, IC50=25μM. 30295477
MDA-MB-468 Cytotoxicity assay 4 hrs Cytotoxicity against human MDA-MB-468 cells incubated for 4 hrs under hypoxic condition followed by compound washout and measured after 5 days by SRB assay, IC50=0.0062μM. 30885680
SW620 Cytotoxicity assay 4 hrs Cytotoxicity against human SW620 cells incubated for 4 hrs under hypoxic condition followed by compound washout and measured after 5 days by SRB assay, IC50=0.052μM. 30885680
MDA-MB-468 Cytotoxicity assay 4 hrs Cytotoxicity against human MDA-MB-468 cells incubated for 4 hrs under aerobic condition followed by compound washout and measured after 5 days by SRB assay, IC50=4.403μM. 30885680
SW620 Cytotoxicity assay 4 hrs Cytotoxicity against human SW620 cells incubated for 4 hrs under aerobic condition followed by compound washout and measured after 5 days by SRB assay, IC50=20.6μM. 30885680
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Información química, almacenamiento y estabilidad (Chemical Information, Storage & Stability)

Peso molecular 449.04 Fórmula

C9H16Br2N5O4P

 Almacenamiento (Desde la fecha de recepción)
Nº CAS 918633-87-1 Descargar SDF Almacenamiento de soluciones madre

Solubilidad (Solubility)

In vitro
Lote:

DMSO : 90 mg/mL (200.42 mM)
(El DMSO contaminado con humedad puede reducir la solubilidad. Usar DMSO fresco y anhidro.)

Ethanol : 90 mg/mL

Water : 10 mg/mL

Calculadora de Molaridad

Masa Concentración Volumen Peso molecular
Calculadora de Dilución Calculadora de Peso Molecular

In vivo
Lote:

Calculadora de formulación in vivo (Solución clara)

Paso 1: Introduzca la información a continuación (Recomendado: Un animal adicional para tener en cuenta la pérdida durante el experimento)

mg/kg g μL

Paso 2: Introduzca la formulación in vivo (Esto es solo la calculadora, no la formulación. Por favor, contáctenos primero si no hay una formulación in vivo en la sección de Solubilidad.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Resultados del cálculo:

Concentración de trabajo: mg/ml;

Método para preparar el líquido maestro de DMSO: mg fármaco predissuelto en μL DMSO ( Concentración del líquido maestro mg/mL, Por favor, contáctenos primero si la concentración excede la solubilidad del DMSO del lote del fármaco. )

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadirμL PEG300, mezclar y clarificar, luego añadirμL Tween 80, mezclar y clarificar, luego añadir μL ddH2O, mezclar y clarificar.

Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadir μL Aceite de maíz, mezclar y clarificar.

Nota: 1. Por favor, asegúrese de que el líquido esté claro antes de añadir el siguiente disolvente.
2. Asegúrese de añadir el (los) disolvente(s) en orden. Debe asegurarse de que la solución obtenida, en la adición anterior, sea una solución clara antes de proceder a añadir el siguiente disolvente. Se pueden utilizar métodos físicos como el vórtice, el ultrasonido o el baño de agua caliente para ayudar a la disolución.

Mecanismo de acción (Mechanism of Action)

In vitro

Evofosfamide (TH-302) is selectively potent under hypoxia and stable to liver microsomes. Substitution of the chlorine with bromine on the phosphorus mustard in 3b increases the potency by 10-fold and maintaines the high hypoxic selectivity [Hypoxia cytotoxicity ratio (HCR) = 270]. In both human lung cancer H460 cells and human colon cancer HT29 cells, potent cytotoxicity of this compound is observed under N2. It inhibits H460 cells and HT29 cells with IC90 of 0.1 μM and 0.2 μM, respectively. It shows much enhanced potency in H460 spheroids compared to H460 monolayer cells under normoxia. This compound exhibits potent cytotoxicity to MM cells with hypoxic selectivity and dose dependency. It can induce G0/G1 cell-cycle arrest under hypoxic conditions. The effect of it on cell-cycle machinery is mediated by down-regulating cyclin D1/2/3, CDK4/6, p21cip-1, p27kip-1, and pRb expression, whereas CDK2 expression remained undisturbed. It can induce dose-dependent apoptosis in both human and murine MM cells in hypoxic conditions. TH-302-activated apoptosis is mediated through down-regulating the antiapoptotic proteins BCL-2 and BCL-xL, as well as up-regulating the expression of cleaved proapoptotic protein caspase-3, -8, and -9 and poly ADP-ribose polymerase. In contrast to the hypoxia-specific toxicity, it shows very low toxicity in normoxic condition, even at high concentrations.

In vivo

Evofosfamide (TH-302) inhibits primary tumor growth by 41% on day 25 after implantation, whereas this compound plus a nucleoside analog inhibits primary tumor growth by 96% on day 25. When administered at 6.25, 12.5, 25, or 50 mg/kg in the H460 NSCLC xenograft model QD × 5/wk × 2 wks (once a day for 5 days per week for 2 weeks) i.p., it shows tumor growth inhibition at Day 22 of 43%, 51%, 75%, and 89%, respectively. At 100 mg/kg, it causes a decrease in blood cell counts 3 days after treatment end, but these are totally recovered 7 days post-treatment. Under all tested regimens, it exhibits efficacy metrics ranging from 58% to 89% tumor growth inhibition. Its induced cell killing is breathing oxygen concentration dependent, with the greatest cytotoxicity occurring when the tumor-bearing mice are exposed to low oxygen concentrations. Tumor growth is significantly reduced by this compound in animals breathing 10% O2 compared with 95% O2 breathing. After treatment, the pimonidazole-positive area is significantly decreased at 48 hours after dosing (6.3 % in vehicle vs. 1.8 % in the TH-302 treatment group).

Referencias
  • [4] https://pubmed.ncbi.nlm.nih.gov/22184053/

Información del ensayo clínico (Clinical Trial Information)

(datos de https://clinicaltrials.gov, actualizado el 2024-05-22)

Número NCT Reclutamiento Condiciones Patrocinador/Colaboradores Fecha de inicio Fases
NCT02020226 Unknown status
Solid Tumors
Threshold Pharmaceuticals
 November 2013 Phase 1
NCT01833546 Completed
Solid Tumor|Pancreatic Cancer
Merck KGaA Darmstadt Germany|Threshold Pharmaceuticals
 April 18 2013 Phase 1