Cat. No.: HY-109747

RO-28-2653

RO-28-2653 is an orally active, blood-brain barrier-penetrant selective MMP-2/MMP-9/MT1-MMP inhibitor and citrate synthase (Citrate synthase) (HY-P2739) inhibitor. RO-28-2653 inhibits rat citrate synthase in a non-competitive manner (K_i = 18-25 µM), blocks MMP-induced vasodilation, and suppresses MT1-MMP catalytic activity-dependent VEGF-A upregulation and VEGF-A mRNA elevation without affecting basal VEGF-A levels. RO-28-2653 reduces cigarette smoke extract-induced VEGF protein, partially inhibits endothelial cell proliferation in co-culture systems, suppresses neovascularization, and exhibits antitumor and antimetastatic activity in mouse tumor models. RO-28-2653 can be used for research on age-related cognitive decline, obesity-induced cognitive impairment, breast cancer, olfactory mucosal tumors, and colon cancer.

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

CAS No.: 261956-22-3
Formula: C₂₆H₂₃N₅O₅
Molecular Weight:485.49

Storage:

Please store the product under the recommended conditions in the Certificate of Analysis.

Data Sheet (290 KB)

RO-28-2653 Chemical Structure

CAS No.: 261956-22-3

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Use of MCE Products

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

Biological Activity
- Description
- IC₅₀ & Target
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References
Calculators
Help & FAQs
Classifications
Bioz

Biological Activity

Description

IC₅₀ & Target

^[4]^[3]^[2]

MMP-2

MMP-9

VEGF-A

All MMP Isoforms :

All VEGFR Isoforms :

In Vitro

Anti-cancerLocal Vascular RegulationInnate ImmunityCarbohydrate MetabolismMitochondrial FunctionMitochondrial Oxidative Phosphorylation

Anti-cancer

Local Vascular Regulation

RO-28-2653 (1 µM; 24 h) inhibits MT1-MMP-driven VEGF-A mRNA upregulation in MCF7 MT/1 and MT/3 clones without affecting basal VEGF-A mRNA in C10 and C11 control clones^[2].
RO-28-2653 (0.1-1 μM; 1 h pre-exposure; 24 h co-culture with CSE) partially reduces CSE-stimulated endothelial cell proliferation in HUVECs co-cultured with SW1116 cells^[5].

Anti-cancer

Local Vascular Regulation

Innate Immunity

RO-28-2653 (0.1-1 μM; 1 h pre-exposure; co-exposure with 100 μg/mL CSE for 5 h) reduces CSE-induced VEGF protein upregulation without altering 5-LOX expression in SW1116 colon cancer cells^[5].

Carbohydrate Metabolism

RO-28-2653 (6.25-50 µM) inhibits rat heart citrate synthase in a non-competitive manner, with a K_i of 25 µM for acetyl coenzyme A and a K_i of 18 µM for oxaloacetic acid (HY-W010382), and this inhibition is attenuated by Triton X-100 and higher mitochondrial protein concentrations^[6].

Local Vascular Regulation

RO-28-2653 (1 μM; 15 min) prevents MMP-2-induced relaxation of phenylephrine (HY-B0769)-precontracted rat IVC, indicating that this venorelaxant effect depends on MMP activity^[4].

Mitochondrial Function

Mitochondrial Oxidative Phosphorylation

RO-28-2653 (10-50 µM; up to 180 min) inhibits state 3 respiration by 23.4% with pyruvic acid (HY-Y0781)/malic acid (HY-Y1311) as substrates at 50 µM, and by 42.5% with succinic acid (HY-N0420) as the substrate, while causing only slight state 4 uncoupling in isolated rat heart mitochondria^[6].
RO-28-2653 (10-50 µM) inhibits citrate synthase in sonicated rat heart mitochondria to 60% and 29% of control at 10 and 50 µM, respectively, without affecting respiratory chain complexes or succinate dehydrogenase^[6].
RO-28-2653 (10-50 µM; up to 180 min) inhibits state 3 respiration by 10.7% at 50 µM with pyruvate/malate as substrate and by 23.5% with succinate as substrate, and causes only slight state 4 uncoupling in saponin-permeabilized rat cardiac fibers^[6].

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

Real Time qPCR^[2]

Cell Line:

human breast adenocarcinoma MCF7 clones (MT/1, MT/3, C10, C11)
Concentration:

1 µM
Incubation Time:

24 h
Result:

Reduced VEGF-A mRNA levels in MT1-MMP-overexpressing MT/1 and MT/3 clones, down-regulating VEGF-A mRNA production to levels comparable to control clones C10 and C11.
Did not affect basal VEGF-A mRNA levels in control clones C10 and C11.

Western Blot Analysis^[5]

Cell Line:

SW1116 human colon adenocarcinoma cells
Concentration:

0.1 and 1 μM
Incubation Time:

1 h pre-exposure; 5 h with 100 μg/mL CSE
Result:

Basal VEGF and 5-LOX protein expression were unaffected at 0.1 and 1 μM.
In CSE-treated SW1116 cells, CSE-induced VEGF protein expression was reduced at 0.1 and 1 μM.
CSE-induced 5-LOX protein expression was unaffected.

Cell Proliferation Assay^[5]

Cell Line:

HUVECs co-cultured non-contact with SW1116 colon cancer cells
Concentration:

0.1 and 1 μM
Incubation Time:

1 h pre-exposure; 24 h co-culture with CSE
Result:

At 0.1 and 1 μM, the CSE-stimulated increase in [³H]-thymidine incorporation was partially inhibited in HUVECs co-cultured with SW1116 cells.

In Vivo

Anti-cancerCarbohydrate MetabolismOthers

Anti-cancer

RO-28-2653 (100 mg/kg/day; p.o. by gavage; daily; 2 months) reduces alachlor-induced olfactory mucosal tumor number by 25% in rats^[3].

Carbohydrate Metabolism

Oral RO-28-2653 (50 mg/kg/day; p.o.; daily; 2 weeks) improves spatial memory and metabolic parameters in HFD-induced obese mice^[1].

Others

Oral RO-28-2653 (50 mg/kg; p.o.; every other day; 4 weeks) improves hippocampus-dependent memory and metabolic dysfunction in aged mice^[1].

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

Animal Model:

C57BL/6J (male, 20 months old at start)^[1]
Dosage:

50 mg/kg
Administration:

p.o.; every other day; 4 weeks
Result:

Improved spatial memory in the forced-alternation Y-maze and enhanced associative memory in contextual fear conditioning in aged mice.
Ameliorated age-related detrimental changes in body weight and lean mass, mitigated impaired glucose metabolism by GTT and ITT, and suppressed increases in serum AST, ALT, and hepatic triglycerides.

Animal Model:

C57BL/6J (male)^[1]
Dosage:

50 mg/kg/day
Administration:

p.o.; daily; 2 weeks
Result:

Improved spatial memory and cognition in obese mice in the object location memory, Y-maze, and water maze tests.
Reduced body weight, blood glucose, triglycerides, and total cholesterol.

Animal Model:

Long-Evans (male, 4-5 weeks of age at study start)^[3]
Dosage:

100 mg/kg/day
Administration:

p.o. by gavage in 0.2% carboxymethylcellulose at 5 mL/kg body weight; daily for 2 months
Result:

Mean tumor number was 3.6 compared with 4.8 in rats receiving alachlor alone, a 25% reduction.
β-Catenin immunoreactivity was detected in olfactory tumors from 3 rats, but not in neoplasms from rats exposed to alachlor alone.

Related Protocols :

Chemical Information

CAS No. 261956-22-3
Molecular Weight 485.49
Formula C₂₆H₂₃N₅O₅
SMILES

O=C1NC(C(N2CCN(C3=CC=C([N+]([O-])=O)C=C3)CC2)(C4=CC=C(C5=CC=CC=C5)C=C4)C(N1)=O)=O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (290 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]. Asthana P, et al. MT1-MMP inhibition rejuvenates ageing brain and rescues cognitive deficits in obesity. Cell discovery. 2025 Sep 23;11(1):76.

[2]. Sounni NE, et al. Up-regulation of vascular endothelial growth factor-A by active membrane-type 1 matrix metalloproteinase through activation of Src-tyrosine kinases. The Journal of biological chemistry. 2004 Apr 02;279(14):13564-74.

[3]. Genter MB, et al. Reduction of alachlor-induced olfactory mucosal neoplasms by the matrix metalloproteinase inhibitor Ro 28-2653. Toxicologic pathology. 2005;33(5):593-9.

[4]. Raffetto JD, et al. MMP-2 induced vein relaxation via inhibition of [Ca2+]e-dependent mechanisms of venous smooth muscle contraction. Role of RGD peptides. The Journal of surgical research. 2010 Apr;159(2):755-64.

[5]. Ye YN, et al. A mechanistic study of colon cancer growth promoted by cigarette smoke extract. European journal of pharmacology. 2005 Sep 05;519(1-2):52-7.

[6]. Opalka JR, et al. Effect of the new matrix metalloproteinase inhibitor RO-28-2653 on mitochondrial function. Biochemical pharmacology. 2002 Feb 15;63(4):725-32.

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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.

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RO-28-2653

All MMP Isoforms

All VEGFR Isoforms