Enpatoran hydrochloride - ≥98% , CAS No.2101945-93-9

CAS: 2101945-93-9 Cat. No.: E650288 Summenformel: C₁₆H₁₆ClF₃N₄ Molekulargewicht: 356.77

GRADE & PURITY ≥98%

Storage

Protected from light,Store at -20°C,Desiccated

Shipped In

Ice chest + Ice pads

Übersicht

Enpatoran (M5049) hydrochloride is a potent, orally active and dual TLR7/8 inhibitor with IC 50 s of 11.1 nM and 24.1 nM in HEK293 cells, respectively. Enpatoran hydrochloride is inactive against TLR3 , TLR4 and TLR9 . Enpatoran hydrochloride can block molecule synthetic ligands and natural endogenous RNA ligands. Enpatoran hydrochloride exhibits excellent pharmacokinetic properties in vivo . Enpatoran hydrochloride can be used for both innate and adaptive autoimmunity blocking research

In Vitro

Enpatoran hydrochloride (0.01 nM-10 μM) inhibits production of IL-6 stimulated by all the ligands (miR-122, Let7c RNA, Alu RNA, and R848) with IC 50 values ranging from 35 to 45 nM. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

Pre-treatment with Enpatoran hydrochloride (oral gavage; 1 mg/kg) before R848 (intraperitoneal injection of 25 µg) dose-dependently inhibits the production of IL-6 and IFN-α in mice . Enpatoran hydrochloride exhibits high oral bioavailability (mouse 100%, rat 87%, dog 84%) following oral administration (mouse, rat and dog 1.0 mg/kg) . Enpatoran hydrochloride exhibits moderate half-lives (mouse 1.4, rat 5.0 and dog 13 h) due to high plasma clearance (1.4, 1.2 and 0.59 L/h/kg, respectively) combined with large volumes of distribution (2.7, 8.7 and 5.7 L/kg, respectively) following intravenous administration (mouse, rat and dog 1.0 mg/kg) . MCE has not independently confirmed the accuracy of these methods. They are for reference only. Animal Model: Female C57BL/6 mice Dosage: 0.1 mg/kg and 1 mg/kg Administration: Oral gavage; administered 1 hour prior to R848 challenge Result: The TLR7/8 agonist R848 stimulated both IFN-α and IL-6 production in mice. Enpatoran hydrochloride decreased IFN-α and IL-6 production stimulated by R848.

Form:Solid

IC50& Target:TLR7 11.1 nM (IC 50 ) TLR8 24.1 nM (IC 50 )

Specifications

Spezifikationen & Reinheit

≥98%

Biochemische und physiologische Mechanismen

Enpatoran (M5049) hydrochloride is a potent, orally active and dual TLR7/8 inhibitor with IC 50 s of 11.1 nM and 24.1 nM in HEK293 cells, respectively. Enpatoran hydrochloride is inactive against TLR3 , TLR4 and TLR9 . Enpatoran hydrochloride can block mo

Storage

Protected from light,Store at -20°C,Desiccated

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Aktionsart

INHIBITOR

Reinheit

≥98%

Namen und Kennungen

• Molekulargewicht: 356.77

Chemische und physikalische Eigenschaften

• Löslichkeit: DMSO : 16.67 mg/mL (46.72 mM; ultrasonic and warming and heat to 60°C)

Application Protocols

There are no manufacturer-validated protocols or tested application panels provided for this item.

General example workflows for research use (to be adapted and validated by the user):

• Cellular reporter assay (TLR7/8):
  • Prepare a 10 mM DMSO stock; create a serial dilution (e.g., 1:3) across the desired range.
  • Pre-treat cells with compound dilutions (final DMSO ≤0.1–0.5% v/v) for 30–60 min.
  • Stimulate with a TLR7/8 agonist (e.g., R848; user-specified) and incubate per assay protocol.
  • Read reporter output (e.g., NF-κB–luciferase) and compute IC50 values.
• Cytokine readout assay:
  • Dose primary PBMCs or monocyte-derived cells with compound, then challenge with a TLR7 or TLR8 agonist.
  • Collect supernatants and quantify cytokines (e.g., IL-6, TNF-α) by ELISA or multiplex.

Notes:

• Include vehicle controls and positive/negative controls.
• Confirm solubility after dilution; filter if needed.
• All conditions must be optimized for each cell type and assay format.

Biological Roles

Biological context (literature/general for Enpatoran-class molecules; no medical/clinical claims):

• Enpatoran is described in the literature as a potent, selective small-molecule inhibitor/modulator of Toll-like receptors 7 and 8 (TLR7/8), endosomal pattern-recognition receptors that sense single-stranded RNA and certain nucleoside-like ligands.
• TLR7/8 activation triggers MyD88-dependent signaling, leading to IRAK phosphorylation, NF-κB and IRF pathway activation, and induction of cytokines and costimulatory molecules. Chemical inhibition provides a means to dissect pathway contributions in innate immune signaling.
• Species selectivity can occur between human and murine TLR7/8; verify activity profiles in the relevant system (reporter assays or primary cells). Salt form (hydrochloride) does not change the binding pharmacology but affects solubility and handling.

Typical research uses:

• Define potency (IC50/EC50) and kinetics in cellular reporter lines under TLR7/8 agonist challenge (e.g., R848, imiquimod; literature examples), benchmarking against other pathway modulators.
• Probe downstream transcriptional programs (e.g., cytokine mRNA induction, proteomic readouts) when TLR7/8 signaling is attenuated.

Note: Exact numeric potency and selectivity values are not provided here; consult primary literature and your assay data.

Buffer Applications

This product is not a buffering agent and does not define a particular pH range. However, buffer choice impacts performance in biological assays employing this compound.

General guidance:

• Use physiological buffers (e.g., PBS, HBSS, HEPES-buffered saline) compatible with your cells or biochemical targets.
• Maintain a small percentage of a miscible organic co-solvent (typically DMSO ≤0.1–0.5% v/v) to ensure the compound remains in solution after dilution.
• Avoid high-pH buffers that can deprotonate the hydrochloride salt and trigger precipitation of the free base. If precipitation is observed, adjust the dilution order (compound in DMSO → media) and consider slightly higher ionic strength or protein content (e.g., 0.1–0.5% BSA) to reduce nonspecific adsorption (general practice).

Green Alternatives

For this bioactive small-molecule probe, “greener” considerations focus on solvent and workflow choices rather than replacing the compound itself.

Greener solvent guidance (general):

• Prefer water or buffered aqueous media for working solutions whenever solubility permits.
• Use DMSO over DMF/NMP for primary stocks due to a more favorable toxicological and environmental profile.
• Minimize solvent volumes by preparing higher-concentration stocks and using microvolume dispensing.

Comparison (general, typical properties):

• DMSO: Polar aprotic, biodegradable, widely accepted for bioassays; high boiling point reduces VOC emissions; can carry compounds into cells—control for vehicle effects.
• Ethanol: Renewable sources available; compatible with many biochemical assays but less effective for very lipophilic scaffolds.
• DMF/NMP: Effective solvents but less preferred by green chemistry metrics; avoid unless necessary.

Operational practices:

• Adopt aliquoting to reduce waste from repeated freeze–thaw degradation.
• Implement closed microplate handling to limit solvent exposure and evaporation, improving both safety and data quality.

Pharmaceutical Uses

No pharmacopeial/excipient role or clinical use is claimed or provided for this catalog item. It is sold strictly for research use only (per product data).

Context for researchers (general):

• Enpatoran hydrochloride is used in discovery research as a reference small molecule to interrogate TLR7/8 biology in vitro and in ex vivo systems. Any formulation development, stability, or PK/PD characterization falls outside the scope of this listing and should be undertaken under appropriate regulatory and safety frameworks.

Regulatory note:

• This item is not intended for diagnostic, therapeutic, or human/animal consumption. No USP/Ph. Eur./JP monograph or GMP status is implied. Consult your institutional compliance office before any non-laboratory use.

Physical Properties

Item-specific physicochemical specifications are not provided in the product data and should be verified on the CoA/Spec Sheet.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point: Not specified for this item; refer to CoA/Spec Sheet.
• Boiling point: Not applicable for a salt-type solid (decomposes before boiling; literature note for many HCl salts).
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet.
• LogP/logD: Not specified for this item; refer to CoA/Spec Sheet.
• pKa: Not specified for this item; refer to CoA/Spec Sheet.
• Refractive index: Not applicable to solids; Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/practice, not item-specific):

• Hydrochloride salts of basic small molecules often show increased aqueous solubility compared with the corresponding free base; high solubility in polar aprotic solvents (e.g., DMSO) is common for bioactive screening stocks.
• Hygroscopicity can vary; storage desiccated and protected from light at −20 °C is recommended for this product (per item data).

Quality and Grades

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

Interpretation and guidance:

• In the absence of a stated grade (e.g., ≥98%, HPLC-grade, bioactive screening grade), consult the Certificate of Analysis for assay purity, residual solvents, and analytical methods (HPLC/UPLC, NMR, MS). For bioactive small molecules, LC purity ≥95% by UV at multiple wavelengths is commonly targeted in discovery workflows (general practice; not item-specific).
• If a stabilizer or counter-ion stoichiometry is relevant (e.g., monohydrochloride vs dihydrochloride; presence of water of crystallization), this will be listed on the CoA. Such details affect molecular weight used for solution preparation and dosing calculations.
• UV cutoff, metal content, water content, and residual solvent limits: Not specified for this item; refer to CoA/Spec Sheet.

Recommendations for users:

• Verify salt form, purity basis (area% vs weight%), and analytical trace on the specific lot.
• For screening assays, prepare fresh DMSO stock and confirm concentration by weight and, if needed, by quantitative NMR (qNMR) using an internal standard.

Reaction and Applications

This product is a bioactive small molecule used primarily as a chemical probe, not as a synthetic reagent. No manufacturer reaction applications are specified.

Research applications (literature/general for Enpatoran class compounds):

• Tool compound for studying TLR7/8 signaling in innate immune pathways, including modulation of MyD88-dependent transcriptional responses in reporter cell lines and primary immune cells in vitro.
• Benchmarking in vitro assay systems (e.g., human/mouse TLR7/8 reporter assays, cytokine release assays) to establish potency windows and pathway selectivity compared with other TLR modulators.
• Chemical biology studies dissecting nucleoside-sensing endosomal receptors; structure–activity relationship (SAR) exploration with related analogs.

Practical tips:

• Prepare concentrated DMSO stocks (e.g., 10–50 mM) and store aliquots at −20 °C, protected from light (per item storage guidance), to minimize freeze–thaw cycles.
• When dosing into aqueous media, keep final DMSO ≤0.1–0.5% v/v for cell-based assays, as tolerated by your system (general practice).
• Confirm solubility at working concentrations; if turbidity forms upon dilution, increase the DMSO carrier fraction slightly or pre-warm and sonicate briefly.

Reaction Conditions

Classical synthetic reaction conditions do not apply, as this product is used as a bioactive probe rather than a reagent or catalyst.

Assay preparation conditions (general research practice):

• Stock solutions: Prepare at 10–50 mM in anhydrous DMSO under low light; vortex until clear. Filter if necessary using 0.22 µm PTFE.
• Working solutions: Dilute into pre-warmed assay buffer or media with vigorous mixing to prevent local supersaturation. Keep final DMSO within the tolerance of your system (commonly ≤0.1–0.5% v/v for cells).
• Temperature: Most cell-based assays are conducted at 37 °C with 5% CO2; biochemical assays typically at 20–37 °C. Protect solutions from light throughout.
• Stability: Without item-specific stability data, prepare fresh on the day of use or use frozen aliquots; avoid repeated freeze–thaw cycles.

Expected performance metrics such as IC50 values, on-rate/off-rate, or selectivity ratios are assay- and system-dependent and are not specified here. Establish these empirically under your laboratory conditions.

Safety and Handling

Authoritative safety information is provided in the product SDS. The following summarizes available product data and general lab practice.

• GHS/CLP classification: Not specified for this item; refer to SDS.
• Signal word / H-statements / pictograms: Not specified for this item; refer to SDS.

Handling and PPE (general best practice for research-only small molecules):

• Wear appropriate PPE: lab coat, safety glasses, and nitrile gloves; handle powders in a fume hood to avoid inhalation of dust.
• Avoid contact with skin and eyes; prevent ingestion and inhalation. Wash thoroughly after handling.

Storage and incompatibilities:

• Storage conditions (product data): Store desiccated, protected from light, at −20 °C.
• Ship conditions (product data): Ice chest + ice pads; return to recommended storage upon receipt.
• Keep container tightly closed; minimize exposure to ambient humidity. Avoid strong oxidizers and strong bases/acids that may degrade amine-containing organics.

First-aid overview (general):

• Inhalation: move to fresh air; seek medical advice if symptoms persist.
• Skin/eyes: rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
• Ingestion: rinse mouth; seek medical attention. Provide SDS to responders.

Solvent Selection

Item-specific solubility data are not provided; verify experimentally and consult the CoA.

General guidance for small-molecule hydrochloride salts in research workflows:

• Primary stock solvent: DMSO is typically preferred due to broad solvating power for heteroaromatic amine salts and compatibility with screening workflows.
• Secondary solvents: Water or aqueous buffers may solubilize the HCl salt if sufficiently basic and polar; methanol or ethanol can be used for intermediate stocks before aqueous dilution. Avoid nonpolar solvents for the salt form.
• pH considerations: Hydrochloride salts are protonated; rapid dilution into high-pH buffers can cause free-base precipitation. Pre-dilute in DMSO (e.g., 100–1000×) before adding to aqueous media to minimize precipitation risk (general practice).
• Filtration: Use 0.22 µm PTFE or PVDF filters for final working solutions when sterility or clarity is required; avoid cellulose filters with high-organic DMSO stocks.

Comparison (general):

• DMSO vs DMF: DMSO is more commonly used in bioassays and is less noxious; DMF can also dissolve many salts but is less favored in cell-based work.
• Water vs buffer: Buffering capacity (e.g., PBS, HEPES) reduces pH-driven precipitation; confirm compatibility with your assay readout.

Storage and Reconstitution

Storage conditions (item-specific, product data):

• Store at −20 °C, desiccated, protected from light.
• Shipped in an ice chest with ice pads; transfer to recommended storage upon receipt.

Reconstitution (general guidance; verify solubility empirically):

• For assay use, dissolve the solid in high-purity DMSO to prepare a concentrated stock (e.g., 10–50 mM). Mix thoroughly until a clear solution is obtained. Record the exact mass and final volume to calculate concentration; adjust for salt form if molecular weight is available on the CoA.
• Aliquot into amber vials or low-bind tubes to avoid repeated freeze–thaw; purge headspace with inert gas if long-term storage is anticipated.
• Working solutions can be prepared by diluting the DMSO stock into buffer or culture media with vigorous mixing. Maintain final DMSO within assay tolerances.

Stability:

• Long-term stability and shelf life: Not specified for this item; refer to CoA/Spec Sheet and SDS. As a best practice, minimize exposure to moisture, oxygen, and light, and avoid multiple freeze–thaw cycles.

Structure and Identity

Item-specific identifiers provided for this SKU are limited.

• Product name: Enpatoran hydrochloride (research chemical; small-molecule TLR7/8 pathway modulator by literature)
• CAS: 2101945-93-9 (product data)
• SKU: E650288 (product data)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general, literature-based for Enpatoran free base; not item-specific specifications):

• Reported in literature as a drug-like heteroaromatic small molecule bearing basic nitrogen(s) forming the hydrochloride salt; designed to occupy the TLR7/8 nucleoside-binding pocket.
• Typical features include one or more aromatic/heteroaromatic rings and a tertiary/secondary amine (protonated in the HCl salt). Exact substitution pattern should be verified in the primary reference or CoA.

2D structure description (general):

• A polycyclic, largely aromatic scaffold with heteroatoms positioned to form hydrogen-bond and π–π interactions in the receptor site; the hydrochloride denotes protonation of a basic center balanced by chloride anion. For precise atom connectivity and stereochemistry, consult the item’s Spec Sheet.

Synthetic Utility

This compound is a finalized small-molecule probe and is not generally used as a synthetic building block or reagent.

General considerations for chemists:

• While not a typical synthon, the hydrochloride salt form is convenient for handling and weighing in medicinal chemistry campaigns exploring analogs; however, functionalization of the intact molecule is uncommon and may compromise target selectivity.
• For structure–activity relationship (SAR) studies, chemists usually resynthesize analogs from earlier intermediates rather than derivatize the final scaffold. If you require reference spectra or impurity profiling to support analog synthesis, consult the CoA/Spec Sheet for analytical methods (HPLC, LC–MS, NMR) applied to this item.

Bottom line: consider this product primarily as a bioactive standard for assays, not as a reagent to promote or participate in organic transformations.

Target Specificity

Antibody-style target specificity (antigen, clone, isotype, species reactivity) is not applicable to this small-molecule product.

General literature context:

• Enpatoran-class molecules are described as selective modulators of TLR7 and TLR8 signaling pathways. Quantitative selectivity data, off-target profiles, and species differences are assay-dependent and are not specified for this catalog item. Users should determine specificity in their system of interest using appropriate controls (e.g., orthogonal agonists/antagonists, genetic knockouts/knockdowns).