MLN-4760

Catalog No.S8940 Batch:S894001

Technical Data

Formula	C₁₉H₂₃Cl₂N₃O₄
Molecular Weight	428.31	CAS No.	305335-31-3
Solubility (25°C)*	In vitro	Water	86 mg/mL (200.78 mM)
		Ethanol	10 mg/mL (23.34 mM)
		DMSO	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

MLN-4760 (compound 16) is a potent and selective inhibitor of human angiotensin-converting enzyme 2 (ACE2) with IC50 of 0.44 nM. MLN-4760 also inhibits bovine carboxypeptidase A (CPDA) with IC50 of 27 μM.

Targets

ACE2 ^[1] (Cell-free assay)	CPDA ^[1] (Cell-free assay)
0.44 nM	27 μM

In vitro

In vitro, conversion from angiotensin II to angiotensin-(1–7) by mouse ACE2 is blocked by MLN-4760 (10 μM). MLN-4760 inhibits both human and mouse ACE2 activity.^[3] In huMNCs, MLN-4760-B (an isomer of MLN-4760) detects 63% ACE2 with 28-fold selectivity over ACE. In huCD34+ cells, MLN-4760-B detects 38% of ACE2 activity with 63-fold selectivity. In mu-heart and muMNCs, MLN-4760-B is 100- and 228-fold selective for ACE2, respectively. In muLin- cells, MLN-4760-B detects 25% ACE2 activity with a pIC50 of 6.3. The racemic mixture and MLN-4760-A shows lower efficacy and poor selectivity for ACE2 in MNCs and mu-heart.^[4]

In vivo

MLN-4760 blocks ACE2 activity, thus significantly reverses the elevation of carotid Ang-(1-7) level and the decline of carotid Ang II level induced by NaHS in mice. The anti-atherosclerotic property of NaHS is also significantly abolished by treatment with MLN-4760.^[2]

Protocol (from reference)

Cell Assay:^{^[4]}	Cell lines mononuclear cells (MNCs), huCD34+ cells, mu-lineage-depleted (muLin-) cells Concentrations 0.01 nM to 300 μM Incubation Time 1 h, 4 h Method In the case of cell lysates, 10 μg of total protein is assayed for activity in a buffer with the following composition: 50mM MES (4-morpholineethanesulphonic acid), 300mM NaCl, 10μM ZnCl2 and 0.01% Triton X-100, pH 6.5. Reaction is initiated by the addition of 5×10^-5 M substrate. Where applicable, recombinant enzymes are used at a concentration of 0.01 μg per reaction. The fluorescence measurements are performed in the black microtiter plates in a total volume of 100 μl. The plates are read using a fluorescence plate reader SpectraMax M5 at an excitation wavelength 320 nm and emission wavelength 405 nm Fluorescence resulting from the substrate hydrolysis increased with time, and achieved maximum by 1 h with recombinant enzyme however with cell/tissue lysates the maximum fluorescence is observed by 4 h of incubation. Therefore fluorescence recorded at 1 and 4 h of reaction time is taken for calculation of percent enzyme inhibition, when using recombinant enzymes and cell/tissue lysates, respectively. ACE2 activity is defined as the ACE2 inhibitor-sensitive fluorescence and ACE activity is defined as the ACE inhibitor, captopril-sensitive fluorescence, and are expressed as percent inhibition. Unless specified, MLN4760 is used in concentration ranging from 0.01 nM to 300 μM.
Animal Study:^{^[2]}	Animal Models C57BL/6J male apoE^−/− mice Dosages 0.5 mg/kg Administration IP

Cell Assay:^{^[4]}

Cell lines
mononuclear cells (MNCs), huCD34+ cells, mu-lineage-depleted (muLin-) cells
Concentrations
0.01 nM to 300 μM
Incubation Time
1 h, 4 h
Method
In the case of cell lysates, 10 μg of total protein is assayed for activity in a buffer with the following composition: 50mM MES (4-morpholineethanesulphonic acid), 300mM NaCl, 10μM ZnCl2 and 0.01% Triton X-100, pH 6.5. Reaction is initiated by the addition of 5×10^-5 M substrate. Where applicable, recombinant enzymes are used at a concentration of 0.01 μg per reaction. The fluorescence measurements are performed in the black microtiter plates in a total volume of 100 μl. The plates are read using a fluorescence plate reader SpectraMax M5 at an excitation wavelength 320 nm and emission wavelength 405 nm Fluorescence resulting from the substrate hydrolysis increased with time, and achieved maximum by 1 h with recombinant enzyme however with cell/tissue lysates the maximum fluorescence is observed by 4 h of incubation. Therefore fluorescence recorded at 1 and 4 h of reaction time is taken for calculation of percent enzyme inhibition, when using recombinant enzymes and cell/tissue lysates, respectively. ACE2 activity is defined as the ACE2 inhibitor-sensitive fluorescence and ACE activity is defined as the ACE inhibitor, captopril-sensitive fluorescence, and are expressed as percent inhibition. Unless specified, MLN4760 is used in concentration ranging from 0.01 nM to 300 μM.

Animal Study:^{^[2]}

Animal Models
C57BL/6J male apoE^−/− mice
Dosages
0.5 mg/kg
Administration
IP

References

[4] Shrinidh Joshi, et al. Eur J Pharmacol. 2016 Mar 5;774:25-33.

+ Expand

Selleck's MLN-4760 has been cited by 1 publication

ACE2 negatively regulates the Warburg effect and suppresses hepatocellular carcinoma progression via reducing ROS-HIF1α activity [ Int J Biol Sci, 2023, 19(8):2613-2629]	PubMed: 37215979

ACE2 negatively regulates the Warburg effect and suppresses hepatocellular carcinoma progression via reducing ROS-HIF1α activity [ Int J Biol Sci, 2023, 19(8):2613-2629]

PubMed: 37215979

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.