Referencia número: HY-B0782

Acetazolamide

15 Cited Publications

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Based on 15 publication(s) in Google Scholar

Acetazolamide is a carbonic anhydrase (CA) IX inhibitor with an IC₅₀ of 30 nM for hCA IX. Acetazolamide has diuretic, antihypertensive and anti-gonococcal activities.

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Pureza: 99.85%
No. CAS: 59-66-5
Fòrmula: C₄H₆N₄O₃S₂
Peso molecular:222.25

Almacenamiento:

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

Acetazolamide Chemical Structure

No. CAS: 59-66-5

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Actividad biológica
- Descripciòn
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
- Ensayo clínico
Chemical Information
Publications (15)
Solvente y solubilidad
- In Vitro
- In Vivo
Protocolo
Pureza y Documentación
Referencias
Help & FAQs
Classifications
Bioz

Actividad biológica

Descripciòn

Acetazolamide is a carbonic anhydrase (CA) IX inhibitor with an IC₅₀ of 30 nM for hCA IX. Acetazolamide has diuretic, antihypertensive and anti-gonococcal activities^[1]^[4]^[5]^[6].

IC₅₀ & Target

CA IX

Ver todos los productos específicos de isoformas Carbonic Anhydrase :

Cellular Effect

Cell Line	Type	Value	Description	References
HeLa	IC₅₀	2.83 μM Compound: AZA	Cytotoxicity against human HeLa cells assessed as reduction in cell viability after 48 hrs under hypoxia condition by MTT assay Cytotoxicity against human HeLa cells assessed as reduction in cell viability after 48 hrs under hypoxia condition by MTT assay	[PMID: 31901743]
HeLa	IC₅₀	7.82 μM Compound: AZA	Cytotoxicity against human HeLa cells assessed as reduction in cell viability after 48 hrs under normaxia condition by MTT assay Cytotoxicity against human HeLa cells assessed as reduction in cell viability after 48 hrs under normaxia condition by MTT assay	[PMID: 31901743]
HT-29	IC₅₀	>100 μM Compound: AAZ	Cytotoxicity against human HT-29 cells after 96 hrs under hypoxic condition by MTT assay Cytotoxicity against human HT-29 cells after 96 hrs under hypoxic condition by MTT assay	[PMID: 29649740]
HT-29	IC₅₀	>100 μM Compound: AAZ	Cytotoxicity against human HT-29 cells after 96 hrs under normoxic condition by MTT assay Cytotoxicity against human HT-29 cells after 96 hrs under normoxic condition by MTT assay	[PMID: 29649740]
HT-29	IC₅₀	53.78 μM Compound: AAZ	Cytotoxicity in human HT-29 cells assessed as reduction in cell viability incubated for 24 hrs under hypoxic condition by MTT assay Cytotoxicity in human HT-29 cells assessed as reduction in cell viability incubated for 24 hrs under hypoxic condition by MTT assay	[PMID: 31542715]
MDA-MB-231	IC₅₀	>100 μM Compound: AAZ	Cytotoxicity against human MDA-MB-231 cells after 96 hrs under hypoxic condition by MTT assay Cytotoxicity against human MDA-MB-231 cells after 96 hrs under hypoxic condition by MTT assay	[PMID: 29649740]
MDA-MB-231	IC₅₀	>100 μM Compound: AAZ	Cytotoxicity against human MDA-MB-231 cells after 96 hrs under normoxic condition by MTT assay Cytotoxicity against human MDA-MB-231 cells after 96 hrs under normoxic condition by MTT assay	[PMID: 29649740]
Oocyte	IC₅₀	0.86 μM Compound: 1	Inhibition of human aquaporin 4 M23 isoform expressed in Xenopus laevis oocytes Inhibition of human aquaporin 4 M23 isoform expressed in Xenopus laevis oocytes	[PMID: 18178093]
Oocyte	IC₅₀	0.9 μM Compound: 1	Inhibition of human AQP4b-mediated water transport in Xenopus oocytes Inhibition of human AQP4b-mediated water transport in Xenopus oocytes	[PMID: 17178220]
WI-38	IC₅₀	14.45 μM Compound: AZA	Cytotoxicity against human WI-38 cells assessed as reduction in cell viability after 48 hrs by MTT assay Cytotoxicity against human WI-38 cells assessed as reduction in cell viability after 48 hrs by MTT assay	[PMID: 31901743]

In Vitro

Acetazolamide also inhibits hCA II with an IC₅₀ of 130 nM^[1].
Acetazolamide (Ace) is a small heteroaromatic sulfonamide that binds to various carbonic anhydrases with high affinity, acting as a carbonic anhydrase (CA) inhibitor^[2].
Compared with the control group, the high Acetazolamide concentration (AceH, 50 nM), Cisplatin (Cis; 1 μg/mL) and Cis combined with the low Acetazolamide concentration (AceL, 10 nM) treatments significantly reduces viability of Hep-2 cells^[2].
Treatment with the Acetazolamide/Cis combination significantly increases the expression levels of P53, as both AceL+Cis and AceH+Cis treatments result in significantly increased P53 protein expression levels compared with the control group. The Ace/Cis combination treatment significantly reduces the bcl-2/bax expression ratio, and increases the expression of caspase-3 protein, compared with the control group. AceL, AceH, Cis and AceL+Cis treatments significantly reduce the bcl-2/bax ratio compared with the control group^[2].
Combined Ace and Cis treatment effectively promotes apoptosis in Hep-2 cells^[2].
Combined treatment with Ace/Cis markedly decreases the expression of AQP1 mRNA in Hep-2 cells. Both AceH and AceL+Cis treatments decrease the expression of aquaporin-1 (AQP1) mRNA in Hep-2 cells compared with the control group^[2].

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

In Vivo

Acetazolamide (40 mg/kg) significantly potentiates the inhibitory effect of MS-275 on tumorigenesis in neuroblastoma (NB) SH-SY5Y xenografts^[3].
Acetazolamide (40 mg/kg) and/or MS-275 treatment reduce expression of HIF1-α and CAIX in NB SH-SY5Y xenograft^[3].
Acetazolamide (40 mg/kg), MS-275 and Acetazolamide+MS-275 reduce expression of mitotic and proliferative markers in NB SH-SY5Y xenografts^[3].
Acetazolamide (50 mg/kg; PO, for 3 days) significantly reduces the gonococcal load in the vagina of infected mice by 90%^[6].

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

Ensayo clínico

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

Chemical Information

No. CAS 59-66-5
Appearance Solid
Peso molecular 222.25
Fòrmula C₄H₆N₄O₃S₂
Color White to off-white
SMILES

CC(NC1=NN=C(S(=O)(N)=O)S1)=O
Envío

Room temperature in continental US; may vary elsewhere.

Almacenamiento

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

Publications (15)

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Journal Impact Factor

ACS Environ Au

Machine Learning-Assisted Recognition of Environmental Sulfur-Containing Chemicals in Nontargeted Mass Spectrometry Analysis of Inadequate Mass Resolution. [Abstract]

2025 Aug 5;5(6):573-582. PMID: 41277996
Antioxidants (Basel)

Thioamide Compound H0802 Enhances Hypoxia Tolerance by Mimicking Hypoxia-Adaptive Reprogramming of Glucose and Oxygen Metabolism. [Abstract]

2026 Apr 22;15(5):525. PMID: 42193148
Antioxidants (Basel)

Redox Regulation of Phosphatase and Tensin Homolog by Bicarbonate and Hydrogen Peroxide: Implication of Peroxymonocarbonate in Cell Signaling. [Abstract]

2024 Apr 17;13(4):473. PMID: 38671920

Acetazolamide purchased from MedChemExpress. Usage Cited in: Antioxidants (Basel). 2024 Apr 17;13(4):473. [Abstract]

Effect of Acetazolamide, a CA inhibitor, on oxidation of PTEN by H₂O₂. HepG2 cells were pre-incubated in HCO₃⁻-free media, supplemented with 0, 0.01, 0.1, 1, and 10 µM Acetazolamide, at 37 °C with 0.1% CO₂ for 4 h. Then, they were changed to stimulation media with same components plus 0.5 mM H₂O₂. After 10 min, the reaction was terminated by NEM-containing lysis buffer. Western blot data with PTEN and β-actin antibodies are shown Acetazolamide considerably impeded PTEN oxidation in a dose-dependent manner.

Acetazolamide purchased from MedChemExpress. Usage Cited in: Antioxidants (Basel). 2024 Apr 17;13(4):473. [Abstract]

Effect of Acetazolamide, a CA inhibitor, on oxidation of PTEN by H₂O₂. HepG2 cells were pre-incubated in HCO₃⁻-free media, supplemented with 0, 0.01, 0.1, 1, and 10 µM Acetazolamide, at 37 °C with 0.1% CO₂ for 4 h. Then, they were changed to stimulation media with same components plus 0.5 mM H₂O₂. After 10 min, the reaction was terminated by NEM-containing lysis buffer. Western blot data with PTEN and β-actin antibodies are shown. Stars are considered as significances.
Cell Rep

Architecture of androgen receptor pathways amplifying glucagon-like peptide-1 insulinotropic action in male pancreatic β cells. [Abstract]

2023 May 17;42(5):112529. PMID: 37200193

Acetazolamide purchased from MedChemExpress. Usage Cited in: Cell Rep. 2023 May 17;42(5):112529. [Abstract]

GSIS was measured via static incubation in male human islets (n = 3–7 donors, 10 IEQ/condition measured in triplicate) cultured for 24 h with vehicle or DHT (10 nM) and then for 40 min in the presence or absence of a carbonic anhydrase inhibitor Acetazolamide (50 μM).
Anal Chem

Exposome-Scale Investigation of Cl-/Br-Containing Chemicals Using High-Resolution Mass Spectrometry, Multistage Machine Learning, and Cloud Computing. [Abstract]

2025 Jun 3;97(21):11099-11109. PMID: 40401576
Anal Chem

Untargeted Defining Protein-Metabolites Interaction Based on Label-Free Kinetic Size Exclusion Chromatography-Mass Spectrometry. [Abstract]

2020 Jun 2;92(11):7657-7665. PMID: 32384235
Plant Cell Environ

Gα Protein Mediates High-Concentration CO2-Induced Stomata Closure Through Interaction With Carbonic Anhydrase to Promote Guard Cell Hydrogen Peroxide Production in Arabidopsis. [Abstract]

2025 Sep;48(9):6704-6721. PMID: 40421746
Int Immunopharmacol

Acetazolamide as an aquaporin 1 inhibitor mitigates rheumatoid arthritis by reducing angiogenesis via the modulation of the FAK-PI3K/Akt signaling pathway. [Abstract]

2025 Jun 17:158:114845. PMID: 40359885
FASEB J

1-Naphthol Drives Liver Steatosis and Fibrosis via the CA3-CD36 Axis Independent of Obesity. [Abstract]

2026 Jun 15;40(11):e71960. PMID: 42201771
J Biol Chem

Aquaporin 5 deficiency suppresses fatty acid oxidation and delays liver regeneration through the transcription factor PPAR. [Abstract]

2025 Mar;301(3):108303. PMID: 39947476
J Pharm Biomed Anal

Screening method of mildronate and over 300 doping agents by reversed-phase liquid chromatography-high resolution mass spectrometry. [Abstract]

2021 Feb 20:195:113870. PMID: 33453569
Biochem Biophys Res Commun

Glucose transports in the ileum: mechanism, regulation and physiological role of ileal glucose absorption. [Abstract]

2025 Sep 19:785:152676. PMID: 41005286
bioRxiv

2023 Sep 8.
Research Square Print

December 16th, 2022.
Elife

Gq activity- and β-arrestin-1 scaffolding-mediated ADGRG2/CFTR coupling are required for male fertility. [Abstract]

2018 Feb 2:7:e33432. PMID: 29393851

Acetazolamide purchased from MedChemExpress. Usage Cited in: Elife. 2018 Feb 2:7:e33432. [Abstract]

Intracellular pH (pHi) of the ligated efferent ductules with or without Acetazolamide(500 μM) from WT mice and Adgrg2^-/Y mice.

Solvente y solubilidad

In Vitro:

DMSO : 50 mg/mL (224.97 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H₂O : < 0.1 mg/mL (insoluble)

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

4.4994 mL

22.4972 mL

44.9944 mL

5 mM

0.8999 mL

4.4994 mL

8.9989 mL

10 mM

0.4499 mL

2.2497 mL

4.4994 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, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

Calculadora de molaridad

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

Mass Mass

Concentration Concentration

Volume Volume

Molecular Weight Molecular Weight

Calculadora de dilución

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

Concentration _(start) Concentration _(start)

Volume _(start) Volume _(start)

Concentration _(final) Concentration _(final)

Volume _(final) Volume _(final)

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 (11.25 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 (11.25 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 (11.25 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: PBS

Solubility: 1.96 mg/mL (8.82 mM); Clear solution; Need ultrasonic and warming and heat to 60°C

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

Protocolo

Cell Assay

Cell Viability Assay^[2]
Cell line: Hep-2 cells and HUVECs
Concentration: 10 nM and 50 nM
Incubation time: 48 h
Assay: The cell viability of Hep-2 cells and HUVECs is measured by MTT assay. Hep-2 cells and HUVECs in logarithmic growth phase are plated in 96-well plates. Following 48 h of drug treatment as indicated, 200 µL MTT (5 mg/mL) is added to each well. Cells are incubated with the MTT solution at 37°C for 4 h. Then, 150 µL DMSO is added for 5 min. The optical density (OD) values are measured at 490 nm with a Versamax Microplate reader.
Note: Combined treatment effectively reduced viability in Hep-2 cells.

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

Animal Administration

In vivo studies^[3]
Animal model: 4-6 weeks-old female NOD/SCID mice
Dosage: 40 mg/kg, intraperitoneal injection, every day for 2 weeks
Administration: Mice are randomized into four groups (5 mice per group). The control and treatment groups receive intraperitoneal injections of vehicle (PBS) or Acetazolamide (40 mg/kg), MS-275 (20 mg/kg) or the combination, respectively, every day for 2 weeks. Experiments are terminated when tumor sizes exceed 2 cm3 in volume or animals show signs of morbidity. Tumor diameters are measured on a daily basis until termination.
Note: Inhibited tumor growth of NB xenografts with significant anti-tumor growth potentiation effect.

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

Pureza y Documentación

Select Batch:

Pureza: 99.98%

Assay: 99.98%

ee.: 99.98%

Ficha de datos (281 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)
Instrucciones de manejo (2659 KB)

Referencias

[1]. Hou Z, et al. Dual-tail approach to discovery of novel carbonic anhydrase IX inhibitors by simultaneously matching the hydrophobic and hydrophilic halves of the active site. Eur J Med Chem. 2017 May 26;132:1-10. [Content Brief]

[2]. Bayat Mokhtari R, et al. Acetazolamide potentiates the anti-tumor potential of HDACi, MS-275, in neuroblastoma. BMC Cancer. 2017 Feb 24;17(1):156. [Content Brief]

[3]. Gao H, et al. Combined treatment with acetazolamide and cisplatin enhances chemosensitivity in laryngeal carcinoma Hep-2 cells. Oncol Lett. 2018 Jun;15(6):9299-9306. [Content Brief]

[4]. Kassamali R, et al. Acetazolamide: a forgotten diuretic agent. Cardiol Rev. 2011 Nov-Dec;19(6):276-8. [Content Brief]

[5]. Jabeen E, et al. Interaction of antihypertensive acetazolamide with nonsteroidal anti-inflammatory drugs. J Photochem Photobiol B. 2013 Aug 5;125:155-63. [Content Brief]

[6]. Abutaleb NS, et al. In vivo efficacy of acetazolamide in a mouse model of Neisseria gonorrhoeae infection. Microb Pathog. 2022 Mar;164:105454. [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, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	4.4994 mL	22.4972 mL	44.9944 mL	112.4859 mL
	5 mM	0.8999 mL	4.4994 mL	8.9989 mL	22.4972 mL
	10 mM	0.4499 mL	2.2497 mL	4.4994 mL	11.2486 mL
	15 mM	0.3000 mL	1.4998 mL	2.9996 mL	7.4991 mL
	20 mM	0.2250 mL	1.1249 mL	2.2497 mL	5.6243 mL
	25 mM	0.1800 mL	0.8999 mL	1.7998 mL	4.4994 mL
	30 mM	0.1500 mL	0.7499 mL	1.4998 mL	3.7495 mL
	40 mM	0.1125 mL	0.5624 mL	1.1249 mL	2.8121 mL
	50 mM	0.0900 mL	0.4499 mL	0.8999 mL	2.2497 mL
	60 mM	0.0750 mL	0.3750 mL	0.7499 mL	1.8748 mL
	80 mM	0.0562 mL	0.2812 mL	0.5624 mL	1.4061 mL
	100 mM	0.0450 mL	0.2250 mL	0.4499 mL	1.1249 mL

Keywords

Acetazolamide

15 Cited Publications

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Acetazolamide

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Publications Citing Use of MedChemExpress (MCE) Acetazolamide

Ver todos los productos específicos de isoformas Carbonic Anhydrase

Actividad biológica

Chemical Information

Publications (15)

Solvente y solubilidad

Protocolo

Pureza y Documentación

Referencias

Help & FAQs

Complete Stock Solution Preparation Table

Acetazolamide Related Classifications

HY-B0782 Related Classifications