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Fig. 1 · Structure
Cat. No.
A10015
CAS No.
90-33-5
Formula
C10H8O3
Mol. Weight
176.2

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4-Methylumbelliferone (4-MU)

Cat. No. A10015 CAS 90-33-5

Hyaluronan (HA) synthesis inhibitor

4-Methylumbelliferone is a selective inhibitor of Hyaluronan (HA) synthesis which is thought to function as an inhibitor via the depletion of UDP-glucuronic acid, the common substrate for HA synthesis.

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Biological Activity

Bioactivity / Mechanism

4-Methylumbelliferone (4-MU) inhibits hyaluronan production in cellular assays. In B16F10 melanoma cells, 30–300 μM 4-MU reduced HA after 48 h. This functional result does not establish direct inhibition of a particular hyaluronan synthase isoform. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

Cellular Effect

Cell Line Activity Value Description Reference
A2780 IC50 > 2 μM Antiproliferative activity against human A2780 cells after 48 hrs by MTT assay [PMID: 25350923]
A549 IC50 > 100 μM Cytotoxicity against human A549 cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
HEK293 IC50 100 μM Inhibition of human recombinant UGT1A1 expressed in HEK293 cells assessed as reduction in bilirubin glucuronidation by LC-MS/MS method [PMID: 21030469]
HEK293 IC50 100 μM Inhibition of human recombinant UGT1A1 expressed in HEK293 cells assessed as reduction in estradiol 3-glucuronidation by LC-MS/MS method [PMID: 21030469]
HepG2 IC50 > 100 μM Cytotoxicity against human HepG2 cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
J774.A1 IC50 > 100 μM Cytotoxicity against mouse J774.A1 cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
MCF7 IC50 54.77 μM Cytotoxicity against human MCF7 cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
NIH3T3 IC50 23.99 μM Cytotoxicity against mouse 3T3 cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
SiHa IC50 > 100 μM Cytotoxicity against human SiHa cells assessed as reduction in cell viability measured after 24 hrs [PMID: 37252094]
SK-MEL-28 IC50 0.58 μM Anticancer activity against human SK-MEL-28 cells assessed as reduction in cell viability incubated for 24 hrs by MTT assay [PMID: 37197457]
SK-MEL3 IC50 1.6 μM Anticancer activity against human SK-MEL3 cells assessed as reduction in cell viability incubated for 24 hrs by MTT assay [PMID: 37197457]
U-937 IC50 > 2000 μM Growth inhibition of human U937 cells by [3H]thymidine incorporation assay [PMID: 18060791]
U-937 CC50 > 2000 μM Cytotoxicity against human U937 cells after 48 hrs by trypan blue assay [PMID: 22925447]
U-937 CC50 > 2000 μM Cytotoxicity against human U937 cells by trypan blue assay after 48 hrs [PMID: 18060791]
U-937 CC50 > 2000 μM Cytotoxicity against human U937 cells by trypan blue assay after 48 hrs [PMID: 18060791]
U-937 IC50 710.9 μM Growth inhibition of human U937 cells by [3H]thymidine incorporation assay [PMID: 18060791]
U-937 IC50 710.9 μM Antiproliferative activity against human U937 cells assessed as incorporation of [3H]-methyl-thymidine after 12 hrs by scintillation counting [PMID: 22925447]

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In Vitro Studies

Hyaluronan reduction in B16F10 cells

Cell-based study · Murine B16F10 melanoma cells [1]

4-MU reduced HA production at each tested concentration after 48 h. This assay does not establish a direct enzyme-binding mechanism or a general cellular potency.

Measured endpoints — not treatment concentrations
EndpointReported resultReference
HA production versus controlReported result Significantly reduced at 30, 100 and 300 μM; p < 0.05[1]

Literature-Reported Treatment Conditions

Treatment concentration
30, 100 and 300 μM
Exposure duration
48 h
Method
Cell-culture exposure in DMEM
Vehicle
DMSO control identified in Fig. 1F; final DMSO percentage not stated in the read methods.
Co-treatment
4-MU alone; 4-MUG was a separate comparator.
Reported limits
Biological replicate count for Fig. 1D and final vehicle percentage not specified in the examined caption/methods.
Methods and source locations

Compound reported in the paper: 4-Methylumbelliferone; Compound label in the paper: 4-MU

Published research on the same compound; no Adooq material validation.

Fig. 1D,F; Results: 4-MUG inhibits HA synthesis in vitro; Experimental procedures: Cell culture, Measurement of HA levels, Statistics

HA production versus control: Modified HA ELISA; samples measured in triplicate; CellTag 700 used for cell normalization. Figure reports mean ± S.E. — Fig. 1D; Measurement of HA levels; Statistics

In Vivo Studies

Mouse studies show that dietary 4-MU exposure is strongly influenced by metabolism and tolerability. With 5% 4-MU chow, circulating HA declined significantly by day 7, and glucuronidated metabolite exposure greatly exceeded parent-drug exposure after two weeks. Initial weight loss preceded resumed weight gain. These findings describe PK, pharmacodynamics and tolerability, rather than a recommended dose or proof of disease efficacy. [2]

Time course of circulating hyaluronan reduction

Animal study · C57BL/6 mice [2]

HA reduction reached significance on day 7, but not at the earlier sampled times. This is a pharmacodynamic observation, not a disease-efficacy endpoint.

Measured endpoints — not treatment concentrations
EndpointReported resultReference
Circulating HA versus control chowReported result Significant decrease on day 7 (P < 0.05); n = 3–5 mice per group[2]

Conditions shared by the study groups

Route
Oral, in chow
Frequency
Ad libitum
Treatment course
7-day sampled time course
Vehicle
Mouse chow
Study groups and reported outcomes
Group / doseComparatorObserved resultReference
4-MU chow versus control chow
5% (w/w) 4-MU in chow		Control chowCirculating HA was significantly lower at day 7.[2]
4-MU chow versus control chow
Model: C57BL/6 mice
Reported limits: Individual intake was not measured for dose assignment; methods estimate approximately 150–250 mg/mouse/day, while Results cites 250 mg/mouse/day. These are estimates, not administered bolus doses.
Methods and source locations

Compound reported in the paper: 4-Methylumbelliferone; Compound label in the paper: 4-MU

Published research on the same compound; no Adooq material validation.

Fig. 1 and corresponding Results; 4-MU treatment; HA quantification; Statistics

Circulating HA versus control chow: HA-specific competitive ELISA-like assay in triplicate; mean ± SEM — Fig. 1 and Results: 4-MU-mediated suppression of HA serum levels requires prolonged treatment

4-MU chow versus control chow · Fig. 1; 4-MU treatment; HA quantification

Parent and metabolite plasma exposure

Animal study · Mice receiving dietary 4-MU; strain and sex not specified for this PK cohort in the examined methods/caption [2]

After two weeks of dietary exposure, glucuronide and sulfate concentrations exceeded parent 4-MU concentrations. Reported averages summarize a 24-h sampling cycle and are not Cmax values.

Measured endpoints — not treatment concentrations
EndpointReported resultReference
4-MU plasma concentrationPlasma concentration 0.89 ± 0.51 μg/ml (5.1 ± 2.9 μM)[2]
4-MUG plasma concentrationPlasma concentration 531 ± 66 μg/ml (1519 ± 189 μM)[2]
4-MUS plasma concentrationPlasma concentration 30.5 ± 2.1 μg/ml (119 ± 8 μM)[2]

Conditions shared by the study groups

Route
Oral, in chow
Frequency
Ad libitum
Treatment course
2 weeks before sampling over 24 h
Vehicle
Mouse chow
Study groups and reported outcomes
Group / doseComparatorObserved resultReference
Dietary 4-MU exposure
5% (w/w) 4-MU in chow		Within-regimen comparison of parent and metabolites; no untreated control needed to interpret reported exposure concentrations4-MUG and 4-MUS concentrations were higher than parent 4-MU (Results: P < 0.001).[2]
Dietary 4-MU exposure
Model: Mouse PK cohort
Reported limits: Individual intake was not measured for dose assignment; methods estimate approximately 150–250 mg/mouse/day, while Results cites 250 mg/mouse/day. These are estimates, not administered bolus doses.
Methods and source locations

Compound reported in the paper: 4-Methylumbelliferone; Compound label in the paper: 4-MU

Published research on the same compound; no Adooq material validation.

Fig. 4b; Results: 4-MU is rapidly metabolized in vivo; Pharmacokinetics of 4-MU; Statistics

4-MU plasma concentration: HPLC–mass spectrometry; 24-h average; mean ± SEM according to Statistics; n = 3 animals per time group — Fig. 4b and accompanying Results; Pharmacokinetics of 4-MU; Statistics

4-MUG plasma concentration: HPLC–mass spectrometry; 24-h average; mean ± SEM according to Statistics; n = 3 animals per time group — Fig. 4b and accompanying Results; Pharmacokinetics of 4-MU; Statistics

4-MUS plasma concentration: HPLC–mass spectrometry; 24-h average; mean ± SEM according to Statistics; n = 3 animals per time group — Fig. 4b and accompanying Results; Pharmacokinetics of 4-MU; Statistics

Dietary 4-MU exposure · Fig. 4b; Pharmacokinetics of 4-MU; 4-MU treatment

Weight changes during dietary 4-MU treatment

Animal study · Healthy C57BL/6 and BALB/c mice [2]

Mice initially lost weight and then resumed weight gain after approximately one week. Resumed weight gain did not mean recovery to control body weights during the displayed follow-up.

Measured endpoints — not treatment concentrations
EndpointReported resultReference
Body weight versus control chowReported result Initial loss, followed by resumed gain after approximately 1 week; n = 7–12 animals per group[2]

Conditions shared by the study groups

Route
Oral, in chow
Frequency
Ad libitum
Treatment course
From 9 weeks to 16–17 weeks of age
Vehicle
Mouse chow
Study groups and reported outcomes
Group / doseComparatorObserved resultReference
4-MU chow in two healthy mouse strains
5% (w/w) 4-MU in chow		Control chowInitial weight loss occurred in both strains; C57BL/6 mice showed a greater decrease.[2]
4-MU chow in two healthy mouse strains
Model: C57BL/6 and BALB/c mice
Reported limits: Individual intake was not measured for dose assignment; methods estimate approximately 150–250 mg/mouse/day, while Results cites 250 mg/mouse/day. These are estimates, not administered bolus doses.
Methods and source locations

Compound reported in the paper: 4-Methylumbelliferone; Compound label in the paper: 4-MU

Published research on the same compound; no Adooq material validation.

Fig. 2a–b; Results: 4-MU induces temporary weight loss; 4-MU treatment; Weight and diabetes monitoring; Statistics

Body weight versus control chow: Body-weight monitoring; Fig. 2a–b reports mean ± SEM and P < 0.05 at marked comparisons — Fig. 2a–b and accompanying Results; Statistics

4-MU chow in two healthy mouse strains · Fig. 2a–b; Results: 4-MU induces temporary weight loss; 4-MU treatment

Specifications

Manufacturer Adooq Bioscience
Actions Inhibitor
CAS Number 90-33-5
Applications Hyaluronan (HA) synthesis inhibitor
Formula C10H8O3
Molecular Weight 176.2
Synonyms 4-MU
SMILES
CC1=CC(=O)OC2=C1C=CC(=C2)O

Storage & Solubility

In Vitro Solubility

Reported Solubility

SolventReported Solubility
DMSO 30 mg/mL (170.29 mM)
Water Insoluble
Ethanol 30 mg/mL (170.29 mM)

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

  1. [1] Nagy N, Gurevich I, Kuipers HF, et al. 4-Methylumbelliferyl glucuronide contributes to hyaluronan synthesis inhibition. The Journal of biological chemistry (2019). PMID: 30914479 DOI: 10.1074/jbc.ra118.006166 Source material
  2. [2] Kuipers HF, Nagy N, Ruppert SM, et al. The pharmacokinetics and dosing of oral 4-methylumbelliferone for inhibition of hyaluronan synthesis in mice. Clinical and experimental immunology (2016). PMID: 27218304 DOI: 10.1111/cei.12815 Source material

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