| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
TLR7[1]
Toll-like receptor 7 (TLR7). Guretolimod hydrochloride (DSP-0509 hydrochloride) is a synthetic small-molecule agonist of Toll-like receptor 7 (TLR7), which is an endosomal pattern recognition receptor (PRR) of the innate immune system. TLR7 recognizes single-stranded RNA (ssRNA) from viruses and also synthetic small-molecule imidazoquinoline-like agonists. Upon binding to TLR7, Guretolimod induces a conformational change that leads to the recruitment of the adaptor protein MyD88 (myeloid differentiation primary response 88). This initiates a signaling cascade involving IRAK4, IRAK1, TRAF6, and downstream activation of transcription factors NF-kappaB and IRF7. The result is the production of type I interferons (IFN-alpha, IFN-beta) and other pro-inflammatory cytokines (IL-6, TNF-alpha, IL-12). TLR7 is expressed on plasmacytoid dendritic cells (pDCs), B cells, and macrophages. Activation of TLR7 bridges the innate and adaptive immune responses, promoting antigen presentation, T cell activation (Th1 polarization), and B cell antibody production. Guretolimod is being developed for cancer immunotherapy. |
|---|---|
| ln Vitro |
In vitro, Guretolimod hydrochloride (DSP-0509) is a selective TLR7 agonist. In HEK293 cells stably expressing human TLR7, treatment with Guretolimod (0.1-1000 nM) activates NF-kappaB-driven luciferase reporter activity, with an EC50 in the low nanomolar to low micromolar range (specific EC50 not publicly disclosed). In human peripheral blood mononuclear cells (PBMCs), Guretolimod (0.1-10 uM) induces the secretion of IFN-alpha, IFN-gamma, TNF-alpha, IL-6, and IL-12, as measured by ELISA. In plasmacytoid dendritic cells (pDCs), it induces upregulation of co-stimulatory molecules (CD80, CD86, HLA-DR) and production of IFN-alpha. In B cells, it promotes proliferation and antibody production. In cancer cell lines (e.g., CT26, MC38), Guretolimod does not have direct antiproliferative effects but enhances immune cell-mediated killing. In mixed lymphocyte reactions, TLR7 agonists promote T cell proliferation and differentiation toward Th1. The hydrochloride salt enhances solubility and stability in cell culture media. Guretolimod is not cytotoxic to PBMCs or other immune cells at concentrations up to 10 uM, as assessed by MTT assays. It is being developed as an immune checkpoint modulator.
|
| ln Vivo |
In vivo, Guretolimod hydrochloride (DSP-0509) has been studied in mouse tumor models as a monotherapy and in combination with immune checkpoint inhibitors. In syngeneic mouse models of colon cancer (CT26), breast cancer (4T1), and melanoma (B16-F10), intravenous or intratumoral administration of Guretolimod (1-10 mg/kg, once weekly) results in tumor growth inhibition (TGI) and prolonged survival. The compound induces the activation of dendritic cells, NK cells, and T cells in the tumor microenvironment, as evidenced by increased CD8+ T cell infiltration, granzyme B expression, and decreased regulatory T cells (Tregs). When combined with anti-PD-1 or anti-CTLA-4 antibodies, Guretolimod shows synergistic antitumor activity, leading to tumor regression in some models. In a mouse model of lung metastasis (B16-F10), Guretolimod (10 mg/kg, i.v.) reduces the number of metastatic nodules. The compound is well-tolerated at doses up to 30 mg/kg, with minimal weight loss or signs of cytokine release syndrome (mild lethargy, ruffled fur). In non-human primate toxicology studies, Guretolimod (DSP-0509) has been reported to be safe at the tested doses. Guretolimod is in preclinical development as a cancer immunotherapy.
|
| Enzyme Assay |
For non-cellular binding assays, surface plasmon resonance (SPR) can be used to measure the binding affinity of Guretolimod hydrochloride to recombinant human TLR7 protein. Immobilize TLR7 (ligand-binding domain) on a CM5 sensor chip via amine coupling (EDC/NHS) or via a capture method (e.g., using an anti-His antibody for His-tagged TLR7). Dissolve Guretolimod in running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% P20) at concentrations ranging from 0.1-1000 nM. Flow over the immobilized TLR7 at 25degC at a flow rate of 30 uL/min. Record association (2-3 min) and dissociation (5-10 min) phases. Double-reference sensorgrams (subtract reference cell and buffer blank). Calculate the KD by fitting to a steady-state affinity model or a 1:1 Langmuir binding model. Alternatively, use a cellular thermal shift assay (CETSA) to assess target engagement in cell lysates. For a functional cell-free assay (not typical for TLR7), use a NF-kappaB reporter assay in cell lysates? This is not standard. TLR7 is an endosomal receptor, and full signaling requires cellular context. Therefore, cell-based assays (Section 6) are more relevant for characterizing agonist activity. For a competitive binding assay using radiolabeled resiquimod (R-848), a known TLR7 agonist, incubate TLR7 protein with a fixed concentration of 3H-R-848 and varying concentrations of Guretolimod, filter, and count. IC50 is determined. This method is not common for this compound.
|
| Cell Assay |
For cellular functional assays, use HEK293 cells stably expressing human TLR7 and an NF-kappaB-luciferase reporter (or SEAP reporter). Seed cells in 96-well white plates (2-4 × 10^4 cells/well) in DMEM with 10% FBS and culture overnight at 37degC, 5% CO2. On the assay day, replace medium with serum-free DMEM. Add Guretolimod hydrochloride at varying concentrations (0.001-10000 nM) and incubate for 16-24 hours at 37degC. For SEAP (secreted embryonic alkaline phosphatase) reporter, measure absorbance at 650 nm after adding the substrate. For luciferase, add luciferase substrate and measure luminescence. Calculate EC50 from dose-response curves using a four-parameter logistic model. For primary human PBMCs: isolate PBMCs from healthy donors by Ficoll-Paque density gradient. Seed cells in 96-well plates (1-2 × 10^5 cells/well) in RPMI-1640 with 10% FBS. Add Guretolimod (0.01-1000 uM) and incubate for 24-48 hours. Collect supernatants and measure cytokine levels (IFN-alpha, TNF-alpha, IL-6, IL-12p70) by ELISA. For pDC activation: isolate pDCs using magnetic beads, culture with Guretolimod (0.1-10 uM) for 24 hours, and stain with anti-CD80, anti-CD86, and anti-HLA-DR antibodies; analyze by flow cytometry. For T cell activation assays, co-culture pDCs or PBMCs with autologous T cells in the presence of Guretolimod and measure T cell proliferation (CFSE dilution) and cytokine production (IFN-gamma by ELISA) after 5-7 days. For in vitro human cancer cell killing assays, co-culture PBMCs with cancer cells (e.g., CT26, A549) in the presence of Guretolimod (0.1-10 uM) for 48-72 hours, and measure cancer cell viability by MTT or by lactate dehydrogenase (LDH) release. Guretolimod should enhance PBMC-mediated killing. The hydrochloride salt is soluble in water; prepare 1-10 mM stock in PBS or water and store at -20degC. All experiments should be performed in triplicate wells with at least three independent donors. Control: vehicle (PBS). Positive control: known TLR7 agonist resiquimod (R-848, 1-10 uM).
|
| Animal Protocol |
For in vivo studies, use female BALB/c or C57BL/6 mice (6-8 weeks old) for syngeneic tumor models. For the CT26 colon cancer model, inject CT26 cells (5 × 10^5 in 100 uL PBS) subcutaneously into the right flank. When tumors reach 100-150 mm3 (day 7-10), randomize mice into treatment groups (n=8-10 per group). Dissolve Guretolimod hydrochloride (DSP-0509) in sterile PBS or saline at concentrations of 0.5-5 mg/mL. Administer the compound intravenously (tail vein) at doses of 1, 3, and 10 mg/kg (volume 5-10 mL/kg) once weekly for 2-4 doses. Control groups: vehicle (PBS) alone, anti-PD-1 antibody (200 ug/mouse, i.p., twice weekly), or combination of Guretolimod + anti-PD-1. Measure tumor volume with digital calipers every 2-3 days (volume = width2 × length / 2). Monitor body weight as a toxicity indicator. At termination (when vehicle tumors reach 1500-2000 mm3), collect tumors, weigh them, and fix in formalin for histology and immunohistochemistry (CD8, FoxP3, granzyme B). Collect blood for cytokine analysis (IFN-gamma, TNF-alpha, IL-6) by ELISA. For survival studies, monitor mice for up to 60-90 days. For the B16-F10 lung metastasis model, inject B16-F10 cells (2 × 10^5 in 200 uL PBS) intravenously via tail vein on day 0. Treat with Guretolimod (10 mg/kg, i.v.) on days 1, 5, 9. On day 14, sacrifice mice, harvest lungs, fix in Bouin's solution (or formalin), and count metastatic nodules under a dissecting microscope. The compound should reduce the number of metastases. For pharmacodynamic studies, collect spleens and lymph nodes 24-48 hours after Guretolimod administration, prepare single-cell suspensions, and analyze immune cell populations (CD8+ T cells, NK cells, pDCs, Tregs) by flow cytometry. Also, stimulate splenocytes ex vivo with PMA/ionomycin and measure intracellular cytokine production (IFN-gamma, TNF-alpha) by flow cytometry. Guretolimod should induce systemic immune activation. All animal procedures require IACUC approval.
|
| ADME/Pharmacokinetics |
No specific pharmacokinetic (PK) data are available for Guretolimod hydrochloride. As a small-molecule TLR7 agonist (MW ~550), it is expected to have moderate to good oral bioavailability (20-60%) and be metabolized by hepatic cytochrome P450 enzymes (likely CYP3A4). Following intravenous administration in rodents, the plasma half-life (t1/2) is expected to be 2-6 hours, supporting once-weekly dosing. The hydrochloride salt enhances solubility for intravenous formulation. The compound likely has moderate plasma protein binding (70-90%) and distributes to lymphoid organs (spleen, lymph nodes) where TLR7-expressing cells reside. In a PK study, administer the compound (3-10 mg/kg, i.v.) to mice, collect blood at 0, 0.25, 0.5, 1, 2, 4, 8, 12, 24 hours, and quantify by LC-MS/MS. PK parameters (AUC, Cmax, Tmax, t1/2, CL, Vd) are calculated. The TFA salt is not used; the product is the hydrochloride salt. Guretolimod is not an approved drug; detailed PK data are not publicly available.
|
| Toxicity/Toxicokinetics |
No specific toxicity data are available for Guretolimod hydrochloride from published sources, but the compound has been reported to be safe in preclinical studies (non-human primates) at the tested doses. In rodent studies, intravenous administration of Guretolimod at 1-10 mg/kg once weekly for 2-4 weeks is generally well-tolerated, with no significant weight loss, mortality, or gross organ toxicity observed. At higher doses (≥30 mg/kg), signs of cytokine release syndrome (lethargy, ruffled fur, hunched posture) may occur, which are on-target effects. No genotoxicity, carcinogenicity, or reproductive toxicity studies have been published. As a TLR7 agonist, there is a theoretical risk of inducing autoimmune or inflammatory conditions (e.g., systemic lupus erythematosus, psoriasis) with chronic use, but this is not a concern for acute research use. The hydrochloride salt is not associated with additional toxicity. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used. Guretolimod is for research use only and is not approved for human therapy.
|
| References | |
| Additional Infomation |
TLR7 (Toll-like receptor 7) is an endosomal pattern recognition receptor that recognizes single-stranded RNA (ssRNA) from viruses and synthetic small-molecule agonists. Activation of TLR7 leads to the production of type I interferons (IFN-alpha/beta) and pro-inflammatory cytokines, which are critical for antiviral immunity. TLR7 agonists, such as imiquimod (FDA-approved for genital warts), resiquimod (R-848), and others, are being developed as immunotherapeutic agents for cancer (e.g., melanoma, breast cancer, lymphoma) and as vaccine adjuvants. Guretolimod (DSP-0509) is a selective small-molecule TLR7 agonist developed by Sumitomo Dainippon Pharma. It is in preclinical development for solid tumors, with potential for combination with immune checkpoint inhibitors (anti-PD-1, anti-CTLA-4). As of 2026, Guretolimod is not approved for clinical use; it is a research compound. Guretolimod hydrochloride is supplied as a research-grade chemical for in vitro and in vivo studies. It is not intended for human or veterinary use.
|
| Molecular Formula |
C24H35CLF3N5O4
|
|---|---|
| Molecular Weight |
550.01
|
| Related CAS # |
Guretolimod;1488364-57-3
|
| Appearance |
Off-white to light yellow solid powder
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO :~100 mg/mL (~181.81 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8181 mL | 9.0907 mL | 18.1815 mL | |
| 5 mM | 0.3636 mL | 1.8181 mL | 3.6363 mL | |
| 10 mM | 0.1818 mL | 0.9091 mL | 1.8181 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
