4-Methylumbeliferone Treatment at a Dose of 1.2 g/kg/Day Is Safe for Long-Term Usage in Rats.

4-methylumbelliferone (4MU) has been suggested as a potential therapeutic agent for a wide range of neurological diseases. The current study aimed to evaluate the physiological changes and potential side effects after 10 weeks of 4MU treatment at a dose of 1.2 g/kg/day in healthy rats, and after 2 months of a wash-out period. Our findings revealed downregulation of hyaluronan (HA) and chondroitin sulphate proteoglycans throughout the body, significantly increased bile acids in blood samples in weeks 4 and 7 of the 4MU treatment, as well as increased blood sugars and proteins a few weeks after 4MU administration, and significantly increased interleukins IL10, IL12p70 and IFN gamma after 10 weeks of 4MU treatment. These effects, however, were reversed and no significant difference was observed between control treated and 4MU-treated animals after a 9-week wash-out period.

Oral hymecromone decreases hyaluronan in human study participants.

BACKGROUNDHyaluronan (HA), an extracellular matrix glycosaminoglycan, has been implicated in the pathophysiology of COVID-19 infection, pulmonary hypertension, pulmonary fibrosis, and other diseases, but is not targeted by any approved drugs. We asked whether hymecromone (4-methylumbelliferone [4-MU]), an oral drug approved in Europe for biliary spasm treatment that also inhibits HA in vitro and in animal models, could be repurposed as an inhibitor of HA synthesis in humans.METHODSWe conducted an open-label, single-center, dose-response study of hymecromone in healthy adults. Subjects received hymecromone at 1200 (n = 8), 2400 (n = 9), or 3600 (n = 9) mg/d divided into 3 doses daily, administered orally for 4 days. We assessed safety and tolerability of hymecromone and analyzed HA, 4-MU, and 4-methylumbelliferyl glucuronide (4-MUG; the main metabolite of 4-MU) concentrations in sputum and serum.RESULTSHymecromone was well tolerated up to doses of 3600 mg/d. Both sputum and serum drug concentrations increased in a dose-dependent manner, indicating that higher doses lead to greater exposures. Across all dose arms combined, we observed a significant decrease in sputum HA from baseline after 4 days of treatment. We also observed a decrease in serum HA. Additionally, higher baseline sputum HA levels were associated with a greater decrease in sputum HA.CONCLUSIONAfter 4 days of exposure to oral hymecromone, healthy human subjects experienced a significant reduction in sputum HA levels, indicating this oral therapy may have potential in pulmonary diseases where HA is implicated in pathogenesis.TRIAL REGISTRATIONClinicalTrials.gov NCT02780752.FUNDINGStanford Medicine Catalyst, Stanford SPARK, Stanford Innovative Medicines Accelerator program, NIH training grants 5T32AI052073-14 and T32HL129970.

Dual targeting nanoparticles based on hyaluronic and folic acids as a promising delivery system of the encapsulated 4-Methylumbelliferone (4-MU) against invasiveness of lung cancer in vivo and in vitro.

Lung cancer is the most common cause of cancer death worldwide. Thereby, new treatment strategies as targeting nano-therapy present promising possibilities to control the aggressiveness of lung cancer. Dual CD44 and folate receptors targetable nanocapsule based on folic-polyethylene glycol-hyaluronic (FA-PEG-HA) were fabricated to improve the therapeutic activity of 4-Methylumbelliferone (4-MU) toward lung cancer. In this study, we fabricate 4-MU Nps as a hybrid polymeric (protamine) protein (albumin) nanocapsule, then functionalized by targeting layer to form 4-MU@FA-PEG-HA Nps with encapsulation efficacy 96.15%. The in vitro study of free 4-MU, 4-MU Nps and 4-MU@FA-PEG-HA Nps on A549 lung cancer cells reveal that the 4-MU Nps and 4-MU@FA-PEG-HA Nps were more cytotoxic than free 4-MU on A549 cells. The observed therapeutic activity of 4-MU@FA-PEG-HA Nps on urethane-induced lung cancer model, potentiality revealed a tumor growth inhibition via apoptotic mechanisms and angiogenesis inhibition. The results were supported by Enzyme-linked immunosorbent assay (ELIZA) of transforming growth factors (TGFß1) and serum HA, histopathological analysis as well as immunohistochemical Ki67, CD44, Bcl-2 and caspace-3 staining. Moreover, 4-MU@FA-PEG-HA Nps exhibited a promising safety profile. Hence, it is expected that our developed novel nano-system can be used for potential application on tumor therapy for lung cancer.

4-methylumbelliferone-mediated polarization of M1 macrophages correlate with decreased hepatocellular carcinoma aggressiveness in mice.

Hepatocellular carcinoma (HCC) arises in the setting of advanced liver fibrosis, a dynamic and complex inflammatory disease. The tumor microenvironment (TME) is a mixture of cellular components including cancer cells, cancer stem cells (CSCs), tumor-associated macrophages (TAM), and dendritic cells (DCs), which might drive to tumor progression and resistance to therapies. In this work, we study the effects of 4-methylumbelliferone (4Mu) on TME and how this change could be exploited to promote a potent immune response against HCC. First, we observed that 4Mu therapy induced a switch of hepatic macrophages (Mϕ) towards an M1 type profile, and HCC cells (Hepa129 cells) exposed to conditioned medium (CM) derived from Mϕ treated with 4Mu showed reduced expression of several CSCs markers and aggressiveness. HCC cells incubated with CM derived from Mϕ treated with 4Mu grew in immunosuppressed mice while presented delayed tumor progression in immunocompetent mice. HCC cells treated with 4Mu were more susceptible to phagocytosis by DCs, and when DCs were pulsed with HCC cells previously treated with 4Mu displayed a potent antitumoral effect in therapeutic vaccination protocols. In conclusion, 4Mu has the ability to modulate TME into a less hostile milieu and to potentiate immunotherapeutic strategies against HCC.

Inhibition of Systemic Hyaluronan Synthesis Exacerbates Murine Hepatic Carcinogenesis.

BACKGROUND/AIM: Hyaluronan (HA) is used as a biomarker of liver fibrosis, which is a key risk factor for the development of hepatocellular carcinoma (HCC). We examined the effects of prolonged pharmacological inhibition of HA synthesis on liver carcinogenesis. MATERIALS AND METHODS: Liver tumors were induced in mice by administering 0.03% thioacetamide (TAA) in drinking water over a 12-month period. Animals simultaneously received either a diet containing of an inhibitor of HA synthesis [4-methylumbelliferone (4-MU)], or a control diet. RESULTS: Addition of 4-MU resulted in a significantly higher number of tumors compared to TAA treatment alone. Moreover, addition of 4-MU resulted in a dose-dependent increase in maximum tumor size. CONCLUSION: While local HA suppression has been shown to have an inhibitory effect on HCC in vitro and in tumor cell implantation experiments, the present results indicate that systemic inhibition of HA synthesis by 4-MU supplementation facilitates hepatic carcinogenesis in vivo.

Hymecromone in the treatment of motor disorders of the bile ducts: a multicenter, double-blind, placebo-controlled clinical study.

Biliary dyskinesia is frequently encountered in clinical practice and is characterized by pain during or after meals. The present study was designed to assess the action of hymecromone in patients with motor disorders of the bile ducts. One hundred twenty-three patients (36 men and 87 women) were enrolled in the multicenter double-blind placebo-controlled study. The mean age was 60.3 years +/- 14.2 SD. Diagnosis was dyspepsia in 58 patients, dyskinesia in 59, cholelithiasis in five and hepatopathy in one. The patients were divided into two groups. One group (61 patients) was treated with hymecromone (300 mg tablets at a dosage of 1,200 mg/day, 2 tablets midday and evening) and another group (62 patients) was treated with placebo. Treatment lasted for 14 days. Control of dyspepsia and pain symptoms of biliary origin was more marked and constant with hymecromone than with placebo. By the end of the treatment, patients in the hymecromone group showed a 70.3% reduction in intensity of spontaneous abdominal pain, while the placebo group showed a 43.8% reduction. Hymecromone was well accepted by the patients and judged to be effective by the investigator in 88.5% of patients treated. The possibility of using hymecromone in 300-mg tablets in the treatment of motor disorders of the bile ducts is thus confirmed.

On the mutagenic action of some enzyme immunoassay substrates.

The mutagenic action of six compounds used in ELISA, EMIT and EIA assays, was investigated by means of the fluctuation test, with Klebsiella pneumoniae as a test organism, and the Ames' plate incorporation test using Salmonella typhimurium TA 98, TA 100 and TA 1537. It appears that 2,2'-azino-di(3-ethyl-benzthiazoline sulphonic acid (6)) or ABTS exerts mutagenic action on Klebsiella pneumoniae at a concentration of 11 g/l, on Salmonella typhimurium TA 100 at a top agar concentration of 0.1 g/l, on Salmonella typhimurium TA 98 at 0.2 g/l and on Salmonella typhimurium TA 1537 at 10 g/l. With umbelliferone, mutagenic action was found only with Klebsiella pneumoniae at a concentration of 0.8 g/l or higher. With o-phenylenediamine, strong mutagenic activity was found only with strain TA 98 and metabolic activation at a top agar concentration of 0.001 g/l. With 5-aminosalicylic acid, beta-methylumbelliferone and p-nitrophenyl-phosphate, no mutagenic action was observed.

[Treatment of obesity in patients with changes of carbohydrate metabolism]. / Tratamiento en la obesidad de pacientes con alteración en el metabolismo de los hidratos de carbono

The combination of dextroamphetamines, biguanides and choleretics seems to be a good therapy for obese patients with carbohydrate intolerance, since the ponderary curve and carbohydrate tolerance improve. Results obtained are statistically significant.