SM-06-09
SM-06-09 is a potent, highly selective, orally active tetrazolone-based HDAC6 inhibitor with an IC50 value of 0.49 nM. SM-06-09 promotes tumor-associated macrophage (TAM) polarization toward an antitumor M1-like phenotype and enhances macrophage phagocytosis, antigen presentation, and T-cell activation. SM-06-09 remodels the tumor immune microenvironment, exhibits antitumor activity in melanoma models, and enhances the efficacy of anti-PD-1 immune checkpoint blockade.
For research use only. We do not sell to patients.
- Formula: C16H13N7O4
- Molecular Weight:367.32
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
Biological Activity
Description
IC50 & Target
[1]|
HDAC6 0.49 nM (IC50) |
HDAC10 16.9 nM (IC50) |
HDAC2 173 nM (IC50) |
HDAC3 258 nM (IC50) |
HDAC1 338 nM (IC50) |
HDAC8 453 nM (IC50) |
HDAC7 4443 nM (IC50) |
HDAC5 7602 nM (IC50) |
HDAC4 12415 nM (IC50) |
HDAC9 26035 nM (IC50) |
HDAC11 1891 nM (IC50) |
HDAC6 9.31 (pIC50) |
In Vitro
SM-06-09 (Compound 3m) potently inhibits HDAC6 with an IC50 value of 0.49 nM and exhibits good selectivity over HDAC10 (IC50=16.9 nM), HDAC2 (IC50=173 nM), HDAC3 (IC50=258 nM), HDAC1 (IC50=338 nM), and HDAC8 (IC50=453 nM)[1].
SM-06-09 (0.15-10 μM) dose-dependently inhibits total HDAC activity in RAW264.7 macrophages, achieving approximately 85% inhibition at 10 μM[1].
SM-06-09 (0.5-10 μM; 24 h) increases acetylated α-tubulin levels in RAW264.7 macrophages, confirming cellular HDAC6 inhibition[1].
SM-06-09 (0.15-20 μM; 24 h) exhibits minimal cytotoxicity in RAW264.7 cells, with less than 10% growth inhibition at the highest tested concentration[1].
SM-06-09 (5 μM; 16 h after polarization) promotes M1 polarization in bone marrow-derived macrophages (BMDMs), upregulates Tnf and Nos2 expression, and downregulates Arg1 and Fizz1 expression. SM-06-09 (5 μM) also decreases Arg1 protein levels while maintaining iNOS expression[1].
SM-06-09 (5 μM) induces transcriptional reprogramming in M1- and M2-polarized BMDMs, downregulates M2-associated genes including Arg1 and Retnla, and upregulates pro-inflammatory genes such as Cxcl3, Cxcl5, and C3. SM-06-09 enriches pathways involved in antigen presentation, T-cell activation, and TNF/IFN inflammatory signaling while suppressing pathways associated with extracellular matrix organization and mesenchymal differentiation[1].
SM-06-09 (5 μM) induces sustained HDAC6 inhibition in BMDMs, with elevated acetylated α-tubulin levels persisting for up to 120 h following a single treatment. SM-06-09 enhances macrophage-mediated phagocytosis of SM1 melanoma cells, suppresses migration of M2-like macrophages (2.5 μM; 24 h), and promotes antigen presentation as well as CD8+ T-cell activation and proliferation[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:Mouse bone marrow-derived macrophages (BMDMs)
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Concentration:0.5, 1, 2.5, 5, and 10 μM
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Incubation Time:16 h after polarization
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Result:Increased expression of M1-associated genes Tnf and Nos2 and decreased expression of M2-associated genes Arg1 and Fizz1.
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Cell Line:RAW264.7 macrophages
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Concentration:10, 5, 2.5, 1.25, 0.625, 0.312 and 0.156 μM
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Incubation Time:24 h
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Result:Displayed minimal cytotoxicity (<10%) throughout the tested concentration range.
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Cell Line:UBC-GFP mouse BMDMs and SM1 melanoma cells
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Concentration:2.5 μM pretreatment
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Incubation Time:24 h
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Result:Significantly reduced migration of M2-like macrophages toward SM1 melanoma cells.
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Cell Line:Murine splenic T cells cocultured with BMDMs (M1-polarized)
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Concentration:Effective concentrations consistent with prior macrophage treatments (primarily 5 μM)
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Incubation Time:72 h
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Result:Significantly increased CD8+ T-cell proliferation following coculture with SM-06-09-treated M1 macrophages.
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Cell Line:Mouse bone marrow-derived macrophages (BMDMs)
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Concentration:0.5, 1, 2.5, 5, and 10 μM
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Incubation Time:24 h after polarization
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Result:Reduced Arg1 protein expression and increased or maintained iNOS protein expression.
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Cell Line:Mouse bone marrow-derived macrophages (BMDMs)
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Concentration:0.5, 1, 2.5, 5, and 10 μM
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Incubation Time:Up to 120 h
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Result:Maintained elevated acetylated α-tubulin levels for up to 120 h following a single treatment.
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Cell Line:RAW264.7 macrophages
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Concentration:0.5, 1, 2.5, 5, and 10 μM
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Incubation Time:24 h
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Result:Increased acetylated α-tubulin levels in a dose-dependent manner, indicating effective HDAC6 inhibition.
Did not significantly alter acetyl-histone H3 levels, indicating cellular selectivity toward HDAC6.
In Vivo
SM-06-09 (25, 50, 100 mg/kg; oral gavage; once daily for 2 weeks) shows no obvious body weight loss or organ toxicity in the C57BL/6 SM1 melanoma model, indicating favorable in vivo safety and tolerability[1].
SM-06-09 (100 mg/kg; oral gavage; once daily for 11-25 days) in combination with an anti-PD-1 antibody (10 mg/kg; intraperitoneal injection; twice weekly) significantly enhances antitumor efficacy and improves the tumor immune microenvironment in a C57BL/6 mouse SM1 melanoma model. The combination increases intratumoral CD45+ immune cells and F4/80+ macrophage infiltration, elevates the proportion of M1-like macrophages and the M1/M2 ratio, and enhances CD8+ T cell infiltration, with increased effector memory and central memory CD8+ T cell populations[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:SM1 murine melanoma model (6-8 weeks, female, C57BL/6)[1]
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Dosage:25 mg/kg
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Administration:intraperitoneal iniections (i.p,), every other day for 10-23 days
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Result:Reduced tumor growth significantly compared to the vehicle group.
Reduced the proportion of M2-like macrophages within tumors.
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Animal Model:SM1 murine melanoma model (6-8 weeks, female, C57BL/6)[1]
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Dosage:25, 50, 100 mg/kg
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Administration:Oral gavage (p.o.), daily for 2 weeks
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Result:Did not induce obvious body weight loss or observable organ toxicity.
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Animal Model:SM1 murine melanoma model (6-8 weeks, female, C57BL/6; combined with 10 mg/kg anti-PD-1)[1]
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Dosage:100 mg/kg
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Administration:Oral gavage (p.o.), once daily for 11-25 days
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Result:Produced significantly greater tumor growth inhibition than monotherapies.
Increased infiltration of CD45+ immune cells and F4/80+ macrophages in tumors.
Markedly elevated the percentage of M1-like macrophages and the M1/M2 ratio.
Enhanced CD8+ T cell infiltration with increased effector memory and central memory CD8+ T cells.
Chemical Information
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Molecular Weight 367.32
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Formula C16H13N7O4
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SMILES
O=C(NO)C1=NOC(CCN2C(N(C3=CC(C=CC=C4)=C4N=C3)N=N2)=O)=C1
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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Phagocytosis Functional Assay
A phagocytosis functional assay measures the ability of phagocytic cells, such as neutrophils, macrophages, monocytes, or microglia/macrophages, to bind and internalize particulate targets including bacteria, yeast particles, beads, or myelin particles. Fluorescent flow-cytometry assays detect target uptake as fluorescence associated with gated phagocytes, while pH-sensitive dyes such as pHrodo increase signal in acidic phagosomal compartments and therefore preferentially report internalized particles rather than particles remaining outside the cell. Microscopy or high-content imaging can be used to confirm intracellular localization and, in some protocols, to follow uptake kinetics.
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Subcutaneous Cell-Line-Derived Xenograft
Subcutaneous cell-line-derived xenograft (CDX) models are established by implanting cultured human cancer cell lines into immunodeficient mice, where the injected cells form localized tumors that can be monitored in vivo as a measure of tumorigenic potential, growth kinetics, and treatment response. These models are widely used in oncology research because they allow reproducible tumor formation and enable comparative assessment of tumor growth between different cell lines or genetic manipulations in a controlled in vivo microenvironment. Subcutaneous implantation of cancer cells in immunodeficient mice is a standard approach for evaluating tumor growth behavior and therapeutic response across multiple cancer types, including prostate, esophageal, pancreatic, and colon cancer models.
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Research Protocol for Cancer Immunology
Cancer immunology studies how the immune system recognizes, suppresses, edits, or fails to eliminate malignant cells through tumor antigen release, antigen presentation, T-cell priming, immune trafficking, tumor-cell killing, and feedback inhibition in the tumor microenvironment. The cancer-immunity cycle links tumor antigenicity, dendritic-cell priming, CD8+ T-cell infiltration, cytotoxic function, and immune-checkpoint regulation to tumor rejection or immune escape. Immune-checkpoint pathways such as PD-1/PD-L1 and CTLA-4 suppress antitumor T-cell activity and can be therapeutically blocked, but many tumors remain resistant because of poor antigen presentation, weak T-cell infiltration, suppressive myeloid cells, regulatory T cells, and tumor-intrinsic immune-exclusion programs. Unresolved questions include which immune-cell states predict response, how tumor-intrinsic pathways exclude immune cells, how myeloid suppression limits checkpoint blockade, and which combination strategies
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Keywords
- SM-06-09
- HDAC
- HDAC6 inhibitor
- HDAC10
- tumor-associated macrophages
- TAMs
- macrophage reprogramming
- M1 macrophage polarization
- M2 macrophage
- RAW264.7
- BMDM
- bone marrow-derived macrophages
- C57BL/6 mice
- UBC-GFP mice
- melanoma
- SM1 melanoma
- tumor microenvironment
- TME
- immune checkpoint blockade
- anti-PD-1
- antigen presentation
- phagocytosis
- T-cell activation
- T-cell proliferation
- immunotherapy
- epigenetics
- acetylated α-tubulin
- Arg1
- Fizz1
- iNOS
- TNF-α
- Inhibitor
- inhibitor
- inhibit