Synthesis, Characterization, and in vivo Distribution of Intracellular Delivered Macrolide Short‐Chain Fatty Acid Derivatives

Straß, Simon; Schwamborn, Anna; Keppler, Manuel; Cloos, Natascha; Guezguez, Jamil; Guse, Jan-Hinrich; Burnet, Michael; Laufer, Stefan

doi:10.1002/cmdc.202100139

Cited by 9 publications

(18 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…uptake of Azithromycin into immune cells [25] which we confirmed in a previous study demonstrating beneficial distribution patterns (spleen, liver and lung) for anti-inflammatory effect after oral application [28]. Although some previous studies showed a possible effect of Azithromycin in psoriasis, our focus was on understanding the efficacy of fumarates.…”

supporting

confidence: 76%

See 1 more Smart Citation

Immune cell targeted fumaric esters support a role of GPR109A as a primary target of monomethyl fumarate in vivo

et al. 2023

Self Cite

View full text Add to dashboard Cite

Dimethyl fumarate (DMF) is approved as a treatment for multiple sclerosis (MS), however, its mode of action remains unclear. One hypothesis proposes that Michael addition to thiols by DMF, notably glutathione is immunomodulatory. The alternative proposes that monomethyl fumarate (MMF), the hydrolysis product of DMF, is a ligand to the fatty acid receptor GPR109A found in the lysosomes of immune cells. We prepared esters of MMF and macrolides derived from azithromycin, which were tropic to immune cells by virtue of lysosomal trapping. We tested the effects of these substances in an assay of response to LPS in freshly isolated human peripheral blood mononuclear cells (PBMCs). In this system, we observed that the 4'' ester of MMF (compound 2 and 3) reduced levels of IL-1β, IL-12 and TNFα significantly at a concentration of 1 µM, while DMF required about 25 µM for the same effect. The 2' esters of MMF (compound 1 and 2) were, like MMF itself, inactive in vitro. The 4'' ester formed glutathione conjugates rapidly while the 2' conjugates did not react with thiols but did hydrolyze slowly to release MMF in these cells. We then tested the substances in vivo using the imiquimod/isostearate model of psoriasis where the 2' ester was the most active at 0.06 to 0.12 mg/kg (approximately 0.1 µmol/kg), improving skin score, body weight and cytokine levels (TNFα, IL-17A, IL-17F, IL-6, IL-1β, NLRP3 and IL-23A). In contrast, the thiol reactive 4'' ester was less active than the 2' ester while DMF was ca. 300-fold less active. The thiol reactive 4'' ester was not easily recovered from either plasma or organs while the 2' ester exhibited conventional uptake and elimination. The 2' ester also reduced levels of IL-6 in acute MSU induced inflammation. These data suggest that mechanisms that are relevant in vivo center on the release of MMF. Given that GPR109A is localized to the lysosome, and that lysosomal trapping increases 2' ester activity by >300 fold, these data suggest that GPR109A may be the main target in vivo. In contrast, the effects associated with GSH conjugation in vitro are unlikely to be as effective in vivo due the much lower dose in use which cannot titrate the more concentrated thiols. These data support the case for GPR109A modulation in autoimmune diseases.

show abstract

supporting

confidence: 76%

“…and organs collected 2 and 4 h after treatment. Collected samples stored at -25 °C until further processing, as published [28].…”

Section: Pharmacokinetic Studiesmentioning

confidence: 99%

Immune cell targeted fumaric esters support a role of GPR109A as a primary target of monomethyl fumarate in vivo

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

“…CSY5669 and its metabolites were quantified using liquid chromatography coupled to a triple quadrupole MSMS detector (API 4500, Sciex, Canada) as described previously ( 45 ). Briefly, samples were homogenized in 6 volumes of acetonitrile, and the resulting supernatant was separated via reverse phase chromatography followed by mass-specific detection.…”

Section: Methodsmentioning

confidence: 99%

The Azithromycin Pro-Drug CSY5669 Boosts Bacterial Killing While Attenuating Lung Inflammation Associated with Pneumonia Caused by Methicillin-Resistant Staphylococcus aureus

Saris

Qin

Linge

et al. 2022

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

Antibiotic resistance is a major problem, with methicillin-resistant Staphylococcus aureus (MRSA) being a prototypical example in surgical and community-acquired infections. S. aureus , like many pathogens, is immune evasive and able to multiply within host immune cells. Consequently, compounds that aid host immunity (e.g., by stimulating the host-mediated killing of pathogens) are appealing alternatives or adjuncts to classical antibiotics.

show abstract

“…Frozen organ samples were thawed and treated as described earlier [20]. Briefly, samples were digested with Proteinase K in a buffer volume in µL corresponding to sample weight in mg (Proteinase K, VWR, diluted 1:40 in phosphate buffer 20 mM) for 1 h at 50 °C and homogenized at 6 m/s for 40 s (FastPrep-24 5G, MP Biomedicals).…”

Section: Sample Preparation For Pharmacokinetic Assessmentmentioning

confidence: 99%

Isostearic acid is an active component of imiquimod formulations used to induce psoriaform disease models

et al. 2023

Self Cite

View full text Add to dashboard Cite

Topical imiquimod based creams are indicated as immune stimulants for papillomas and various skin neoplasms. Imiquimod is considered a TLR7 ligand. These creams are also used in research to induce skin inflammation in mice as a model for psoriasis. We observed that this inflammatory response was not strictly imiquimod dependent and we set out to establish which components drive the proinflammatory effects. To this end, we examined the induction response in a BALB/cJRj mouse model, in which 50 mg of cream is applied to 2 cm 2 of skin (125 mg/kg imiquimod -5% W/V, and/or 625 mg/kg isostearic acid -25% W/V). Comparing cream formulations containing isostearic acid, imiquimod and the combination, we observed that isostearic acid causes skin inflammation within 2 days, whereas imiquimod requires up to 5 days for initial signs. Isostearic acid activated an inflammasome response, stimulated release of proinflammatory cytokines and upregulated the IL-23/17 axis. Animals treated with isostearic acid had enlarged livers (+40% weight), which was not observed with imiquimod alone. Imiquimod was readily metabolized and cleared from plasma and liver, but was maintained at high levels in the skin throughout the body (200 mM at area of application; 200 µM in untreated skin). Imiquimod application was associated with splenomegaly, cytokine induction/release and initial body weight loss over 3 days. Despite high imiquimod skin levels throughout the animal, inflammation was only apparent in the treated areas and was less severe than in isostearic acid groups. As the concentrations in these areas are well above the 10 µM required for TLR7 responses in vitro, there is an implication that skin inflammation following imiquimod is due to effects other than TLR7 agonism (e.g., adenosine receptor agonism). In brain, isostearic caused no major changes in cytokine expression while imiquimod alone sightly stimulated expression of IL-1β and CCL9. However, the combination of both caused brain induction of CCL3, -9, CXCL10, -13, IL-1β and TNFα. The implication of these data is that isostearic acid facilitates the entry of imiquimod or peripherally secreted cytokines into the brain. Our data suggest that psoriaform skin responses in mice are more driven by isostearic acid, than generally reported and that the dose and route used in the model, leads to profound systemic effects, which may complicate the interpretation of drug effects in this model.

show abstract

Synthesis, Characterization, and in vivo Distribution of Intracellular Delivered Macrolide Short‐Chain Fatty Acid Derivatives

Cited by 9 publications

References 68 publications

Immune cell targeted fumaric esters support a role of GPR109A as a primary target of monomethyl fumarate in vivo

Immune cell targeted fumaric esters support a role of GPR109A as a primary target of monomethyl fumarate in vivo

The Azithromycin Pro-Drug CSY5669 Boosts Bacterial Killing While Attenuating Lung Inflammation Associated with Pneumonia Caused by Methicillin-Resistant Staphylococcus aureus

Isostearic acid is an active component of imiquimod formulations used to induce psoriaform disease models

Contact Info

Product

Resources

About