Cat. No.: HY-10535

Evofosfamide

Name: Evofosfamide
Brand: MedChemExpress
SKU: HY-10535
Price: 140.00 USD
Availability: InStock
Rating: 5 (8 reviews)

(Synonyms: TH-302)

8 Cited Publications

Customer Review

Based on 8 publication(s) in Google Scholar

Evofosfamide (TH-302) is a hypoxia-activated proagent with IC₅₀ of 10 μM and 1000 μM in hypoxia (N₂) and normoxia (21% O₂), respectively.

For research use only. We do not sell to patients.

Purity : 98.0%
CAS No.: 918633-87-1
Formula: C₉H₁₆Br₂N₅O₄P
Molecular Weight:449.04

Storage:

Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month

Evofosfamide Chemical Structure

CAS No.: 918633-87-1

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Top Publications Citing

Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
- Clinical Trial
Chemical Information
Publications (8)
Solvent & Solubility
- In Vitro
- In Vivo
Protocols (1)
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

Evofosfamide (TH-302) is a hypoxia-activated proagent with IC₅₀ of 10 μM and 1000 μM in hypoxia (N₂) and normoxia (21% O₂), respectively.

IC₅₀ & Target

Hypoxia-activated prodrug^[1]

Cellular Effect

Cell Line	Type	Value	Description	References
DU-145	IC₅₀	4.14 μM Compound: 2; TH-302	Cytotoxicity against human DU145 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay Cytotoxicity against human DU145 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay	[PMID: 29259746]
DU-145	IC₅₀	842 μM Compound: 2; TH-302	Cytotoxicity against human DU145 cells incubated for 2 hrs under normoxic condition measured after 72 hrs by Alamar blue assay Cytotoxicity against human DU145 cells incubated for 2 hrs under normoxic condition measured after 72 hrs by Alamar blue assay	[PMID: 29259746]
H-EMC-SS	IC₅₀	0.13 μM Compound: TH-302	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 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	[PMID: 30199705]
H-EMC-SS	IC₅₀	2.9 μM Compound: TH-302	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 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	[PMID: 30199705]
HT-29	IC₅₀	>100 μM Compound: TH-302	Cytotoxicity against human HT-29 cells pretreated for 24 hrs under normoxic condition followed by compound washout measured after 72 hrs by MTT assay Cytotoxicity against human HT-29 cells pretreated for 24 hrs under normoxic condition followed by compound washout measured after 72 hrs by MTT assay	[PMID: 28350997]
HT-29	IC₅₀	49.51 μM Compound: TH-302	Cytotoxicity against human HT-29 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay Cytotoxicity against human HT-29 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay	[PMID: 28350997]
MDA-MB-468	IC₅₀	0.0062 μM Compound: TH-302	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 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	[PMID: 30885680]
MDA-MB-468	IC₅₀	4.403 μM Compound: TH-302	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 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	[PMID: 30885680]
NCI-H460	IC₅₀	>100 μM Compound: 266	Antiproliferative activity against human NCI-H460 cells Antiproliferative activity against human NCI-H460 cells	[PMID: 30295477]
NCI-H460	IC₅₀	0.019 μM Compound: 3b, TH-302	Cytotoxicity against human H460 cells under hypoxic condition after 2 hrs by Alamar blue staining assay Cytotoxicity against human H460 cells under hypoxic condition after 2 hrs by Alamar blue staining assay	[PMID: 18257544]
NCI-H460	IC₅₀	2 μM Compound: 266	Antiproliferative activity against human NCI-H460 cells in presence of N2 Antiproliferative activity against human NCI-H460 cells in presence of N2	[PMID: 30295477]
NCI-H460	IC₅₀	25 μM Compound: 266	Antiproliferative activity against human NCI-H460 cells in presence of O2 Antiproliferative activity against human NCI-H460 cells in presence of O2	[PMID: 30295477]
NCI-H460	IC₅₀	5.1 μM Compound: 3b, TH-302	Cytotoxicity against human H460 cells under normoxic condition after 2 hrs by Alamar blue staining assay Cytotoxicity against human H460 cells under normoxic condition after 2 hrs by Alamar blue staining assay	[PMID: 18257544]
NCI-H460	IC₅₀	50.97 μM Compound: TH-302	Cytotoxicity against human NCI-H460 cells pretreated for 24 hrs under normoxic condition followed by compound washout measured after 72 hrs by MTT assay Cytotoxicity against human NCI-H460 cells pretreated for 24 hrs under normoxic condition followed by compound washout measured after 72 hrs by MTT assay	[PMID: 28350997]
NCI-H460	IC₅₀	6650 nM Compound: TH-302	Cytotoxicity against human NCI-H460 cells after 24 hrs under normoxic condition by luminescence-based Cell-Titer Glo assay Cytotoxicity against human NCI-H460 cells after 24 hrs under normoxic condition by luminescence-based Cell-Titer Glo assay	[PMID: 29079474]
NCI-H460	IC₅₀	9.08 μM Compound: TH-302	Cytotoxicity against human NCI-H460 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay Cytotoxicity against human NCI-H460 cells pretreated for 24 hrs under hypoxic condition followed by compound washout measured after 72 hrs by MTT assay	[PMID: 28350997]
NCI-H460	IC₅₀	9.3 nM Compound: TH-302	Cytotoxicity against human NCI-H460 cells after 24 hrs under hypoxic condition by luminescence-based Cell-Titer Glo assay Cytotoxicity against human NCI-H460 cells after 24 hrs under hypoxic condition by luminescence-based Cell-Titer Glo assay	[PMID: 29079474]
PC-3	IC₅₀	6.49 μM Compound: 2; TH-302	Cytotoxicity against human PC3 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay Cytotoxicity against human PC3 cells incubated for 2 hrs under hypoxic condition measured after 72 hrs by Alamar blue assay	[PMID: 29259746]
PC-3	IC₅₀	687 μM Compound: 2; TH-302	Cytotoxicity against human PC3 cells incubated for 2 hrs under normoxic condition measured after 72 hrs by Alamar blue assay Cytotoxicity against human PC3 cells incubated for 2 hrs under normoxic condition measured after 72 hrs by Alamar blue assay	[PMID: 29259746]
SW-620	IC₅₀	0.052 μM Compound: TH-302	Cytotoxicity against human SW620 cells incubated for 4 hrs under hypoxic condition followed by compound washout and measured after 5 days by SRB assay Cytotoxicity against human SW620 cells incubated for 4 hrs under hypoxic condition followed by compound washout and measured after 5 days by SRB assay	[PMID: 30885680]
SW-620	IC₅₀	20.6 μM Compound: TH-302	Cytotoxicity against human SW620 cells incubated for 4 hrs under aerobic condition followed by compound washout and measured after 5 days by SRB assay Cytotoxicity against human SW620 cells incubated for 4 hrs under aerobic condition followed by compound washout and measured after 5 days by SRB assay	[PMID: 30885680]

In Vitro

Evofosfamide (TH-302) induces γH2AX and apoptosis. Evofosfamide displays hypoxia-selective and concentration-dependent cytotoxic activity that is comparable in both p53-proficient and -deficient cells. Treatment with Evofosfamide (TH-302) alone causes an accumulation of G₂/M cells. Inhibition of Chk1 by PF47736 in cells treated with Evofosfamide reduces Evofosfamide (TH-302)-mediated G₂/M arrest under both normoxia and hypoxia^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.

In Vivo

Evofosfamide (TH-302) is a hypoxia-activated prodrug known to activate selectively under the hypoxic conditions commonly found in solid tumors. The mean values of normalized K^trans decrease 69.2% for Evofosfamide (TH-302)-treated mice in Hs766t tumors, decrease 46.1% for Mia PaCa-2 tumors and increase 4.9% in SU.86.86 tumors. Both changes for Hs766t and Mia PaCa-2 treatment groups are statistically significant (P<0.01) when compare to their own control group^[2]. A significant reduction in the hypoxic fraction (HF) to 2.1%±4.7% is seen after 95% oxygen breathing (P<0.001), whereas 7% oxygen breathing significantly increase the HF to 29.5%±14.7% (P=0.029). Exposing rhabdomyosarcoma-bearing rats to increasing oxygen conditions abolish the effect of TH-302 and reduce the T4×SV from 20.4±3.5 to 15.3±2.5 days (P=0.007), whereas control animals have an increased T4×SV. Upon combination with radiotherapy, the T4×SV of TH-302-treated tumors decrease from 30.8±5.9 (Evofosfamide (TH-302)+radiotherapy) to 25.7±2.9 days (Evofosfamide (TH-302)+radiotherapy+95% O₂)^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial

NCT Number	Sponsor	Condition	Start Date	Phase A549
NCT01329991	Plexxikon\|		2011-05	PHASE1

Chemical Information

CAS No. 918633-87-1
Appearance Solid
Molecular Weight 449.04
Formula C₉H₁₆Br₂N₅O₄P
Color White to yellow
SMILES

O=P(NCCBr)(OCC1=CN=C(N1C)[N+]([O-])=O)NCCBr
Synonyms

TH-302
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

Publications (8)

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Journal Impact Factor

J Immunother Cancer

Neovascular pruning by IDO1 inhibitors can potentiate immunogenic cytotoxicity of ischemia-targeted agents to synergistically enhance anti-PD-1 responsiveness. [Abstract]

2025 May 30;13(5):e011398. PMID: 40447318
Anal Chem

A Dual-Channel Self-Calibrated Fluorescent Probe for the Correlation of Nitroreductase Activity and DNA Damage. [Abstract]

2026 May 24. PMID: 42178687
ACS Med Chem Lett

High-Throughput Screening of Patient-Derived Cultures Reveals Potential for Precision Medicine in Glioblastoma. [Abstract]

2015 Jun 22;6(8):948-52. PMID: 26288699
Front Oncol

Monitoring Treatment Efficacy of Antiangiogenic Therapy Combined With Hypoxia-Activated Prodrugs Online Using Functional MRI. [Abstract]

2021 Apr 30:11:672047. PMID: 33996599
SLAS Discov

Reprint of: Detection and Impact of Hypoxic Regions in Multicellular Tumor Spheroid Cultures formed by Head and Neck Squamous Cell Carcinoma Cells Lines. [Abstract]

2023 Dec 13:100130. PMID: 38101574
SLAS Discov

Detection and impact of hypoxic regions in multicellular tumor spheroid cultures formed by head and neck squamous cell carcinoma cells lines. [Abstract]

2022 Jan;27(1):39-54. PMID: 35058175
Clin Transl Oncol

TH-302 (evofosfamide) monotherapy exerts anticancer activity in Ewing's sarcoma cells under hypoxia. [Abstract]

2025 Jun 14. PMID: 40515777
bioRxiv

2024 Nov 06.

Solvent & Solubility

In Vitro:

DMSO : 94 mg/mL (209.34 mM; Need ultrasonic and warming; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H₂O : 4.35 mg/mL (9.69 mM; ultrasonic and warming and heat to 60°C)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

2.2270 mL

11.1349 mL

22.2697 mL

5 mM

0.4454 mL

2.2270 mL

4.4539 mL

10 mM

0.2227 mL

1.1135 mL

2.2270 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass Mass

Concentration Concentration

Volume Volume

Molecular Weight Molecular Weight

Dilution Calculator

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

Stock Solution Working Solution

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (5.57 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (5.57 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil

Solubility: ≥ 2.5 mg/mL (5.57 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

Working Solution

For the following dissolution methods, please prepare the working solution directly:

It is recommended to prepare fresh solutions and use them promptly within a short period of time.

The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 50% PEG300 50% Saline

Solubility: 10 mg/mL (22.27 mM); Clear solution; Need ultrasonic

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
μL
Number of Animals Number of Animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

%
DMSO +

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
%
+
+
%
Tween-80 + +
%
Saline +

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Working solution concentration: 0.22 mg/mL

Protocols

All Cell Biology

Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (286 KB)
SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Meng F, et al. Enhancement of hypoxia-activated prodrug TH-302 anti-tumor activity by Chk1 inhibition. BMC Cancer. 2015 May 21;15:422. [Content Brief]

[2]. Zhang X, et al. MR Imaging Biomarkers to Monitor Early Response to Hypoxia-Activated Prodrug TH-302 in Pancreatic Cancer Xenografts. PLoS One. 2016 May 26;11(5):e0155289. [Content Brief]

[3]. Peeters SG, et al. TH-302 in Combination with Radiotherapy Enhances the Therapeutic Outcome and Is Associated with Pretreatment [¹⁸F]HX4 Hypoxia PET Imaging. Clin Cancer Res. 2015 Jul 1;21(13):2984-92. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
H₂O / DMSO	1 mM	2.2270 mL	11.1349 mL	22.2697 mL	55.6743 mL
H₂O / DMSO	5 mM	0.4454 mL	2.2270 mL	4.4539 mL	11.1349 mL
DMSO	10 mM	0.2227 mL	1.1135 mL	2.2270 mL	5.5674 mL
	15 mM	0.1485 mL	0.7423 mL	1.4846 mL	3.7116 mL
	20 mM	0.1113 mL	0.5567 mL	1.1135 mL	2.7837 mL
	25 mM	0.0891 mL	0.4454 mL	0.8908 mL	2.2270 mL
	30 mM	0.0742 mL	0.3712 mL	0.7423 mL	1.8558 mL
	40 mM	0.0557 mL	0.2784 mL	0.5567 mL	1.3919 mL
	50 mM	0.0445 mL	0.2227 mL	0.4454 mL	1.1135 mL
	60 mM	0.0371 mL	0.1856 mL	0.3712 mL	0.9279 mL
	80 mM	0.0278 mL	0.1392 mL	0.2784 mL	0.6959 mL
	100 mM	0.0223 mL	0.1113 mL	0.2227 mL	0.5567 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Evofosfamide

Publications Citing Use of MedChemExpress (MCE) Evofosfamide