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Cat. No.: HY-189513

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NSF-951

Name: NSF-951
Brand: MedChemExpress
SKU: HY-189513
Availability: PreOrder

(Synonyms: ZINC67299692)

NSF-951 (ZINC67299692) is a TLR4 agonist (K_d = 16.01 μM). NSF-951 activates the p38 and JNK kinase pathways. NSF-951 induces IL-6 and TNF-α responses and upregulates CD80 and CD86 expression. In mouse immunization studies, NSF-951 alone or in combination with Alum enhances antigen-specific antibody and T cell responses.

For research use only. We do not sell to patients.

CAS No.: 1291868-53-5
Formula: C₂₃H₂₄N₄O₃S
Molecular Weight:436.53

Storage:

Please store the product under the recommended conditions in the Certificate of Analysis.

NSF-951 Chemical Structure

CAS No.: 1291868-53-5

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Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- Cellular Effect
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References
Calculators
Help & FAQs
Classifications
Bioz

Biological Activity

Description

All Toll-like Receptor (TLR) Isoforms :

All p38 MAPK Isoforms :

All JNK Isoforms :

All Interleukin Related Isoforms :

All TNF Receptor Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
RAW264.7	EC50	253.4 nM	Functional potency of NSF-951 for induction of IL-6 production in RAW 264.7 mouse macrophages stimulated for 24 hrs and measured in culture supernatants by sandwich ELISA. Functional potency of NSF-951 for induction of IL-6 production in RAW 264.7 mouse macrophages stimulated for 24 hrs and measured in culture supernatants by sandwich ELISA.	41023411

In Vitro

Innate ImmunityCytokines & ChemokinesAdaptive ImmunityOthers

Innate Immunity

NSF-951 (1-500 µM; 24 h) shows no cytotoxicity in RAW 264.7 mouse macrophages at concentrations up to 200 µM^[1].
NSF-951 (1-500 µM; 24 h) shows no cytotoxicity in THP-1 macrophages at concentrations up to 200 µM^[1].
NSF-951 (1-500 µM; 24 h) is not cytotoxic to BoMac bovine macrophages at concentrations up to 200 µM^[1].
NSF-951 (2 µM; 24 h) activates the p38 and JNK MAP kinase pathways in RAW 264.7 macrophages^[1].

Cytokines & Chemokines

Innate Immunity

NSF-951 (0.2-20 µM; 24 h) is a potent immunostimulatory molecule that induces pro-inflammatory cytokine production in RAW 264.7 mouse macrophages with an EC₅₀ of 253.4 nM^[1].

Adaptive Immunity

Innate Immunity

NSF-951 (2 µM; 24 h) induces the maturation and activation of RAW 264.7 macrophages by upregulating the costimulatory molecules CD80 and CD86 and the maturation marker MHC-II^[1].

Others

NSF-951 binds to the TLR4-MD2 complex of mouse and bovine species with favorable binding energy, and its binding to mTLR4-MD2 is regulated by specific residues including Y122, I68, L101, and L142^[1].
NSF-951 (0.244-500 µM) shows direct binding to the mouse TLR4 ectodomain with a K_D of 16.01 µM^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Cytotoxicity Assay^[1]

Cell Line:

RAW 264.7 mouse macrophage cell line
Concentration:

1-500 µM
Incubation Time:

24 h
Result:

Did not exhibit significant toxicity in the RAW 264.7 cell line at concentrations up to 200 µM.

Cell Cytotoxicity Assay^[1]

Cell Line:

THP-1 human macrophage cell line
Concentration:

1-500 µM
Incubation Time:

24 h
Result:

Did not exhibit significant toxicity in the THP-1 cell line at concentrations up to 200 µM.

Cell Cytotoxicity Assay^[1]

Cell Line:

BoMac bovine macrophage cell line
Concentration:

1-500 µM
Incubation Time:

24 h
Result:

Did not exhibit significant toxicity in the BoMac cell line at concentrations up to 200 µM.

ELISA Assay^[1]

Cell Line:

RAW 264.7 mouse macrophage cell line
Concentration:

0.2-20 µM
Incubation Time:

24 h
Result:

Induced significant, dose-dependent levels of IL-6 and TNF-α in RAW 264.7 macrophages.
The functional potency for IL-6 induction was determined with an EC50 value of 253.4 nM.

Western Blot Analysis^[1]

Cell Line:

RAW 264.7 mouse macrophages
Concentration:

2 µM
Incubation Time:

24 h
Result:

Induced strong phosphorylation of p38 and JNK in RAW 264.7 macrophages.

In Vivo

Adaptive ImmunityCytokines & Chemokines

Adaptive Immunity

Cytokines & Chemokines

NSF-951 (5 µg/mouse; s.c.; once then boosted on day 21) significantly enhances OVA-specific IgG production and T-cell responses (IL-4 and IFN-γ) in immunized mice^[1].
NSF-951 (5 µg/mouse; s.c.; once then boosted on day 21) in AF007 acts as a strong co-adjuvant, amplifying the adjuvant effect of Alum and inducing a Th1-skewed immune response comparable to AS04^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:

C57BL/6 (male, 5-6 weeks old, OVA immunization model)^[1]
Dosage:

5 µg/mouse
Administration:

s.c.; once then boosted on day 21
Result:

Induced significantly higher levels of OVA-specific IgG compared to OVA-only group.
Induced significantly higher secretion of IL-4 and IFN-γ cytokines compared to OVA-only group.

Animal Model:

C57BL/6 (male, 5-6 weeks old, OVA immunization model)^[1]
Dosage:

5 µg/mouse (NSF-951); 500 µg/mouse (Alum)
Administration:

s.c.; once then boosted on day 21
Result:

Generated significantly higher antibody levels compared to OVA-Alum group.
Induced elevated levels of IgG1 and IgG2c.
Enhanced IL-4 levels and induced significantly higher IFN-γ levels, indicating a Th1-skewed response.

Related Protocols :

Chemical Information

CAS No. 1291868-53-5
Molecular Weight 436.53
Formula C₂₃H₂₄N₄O₃S
SMILES

O=C(CSC1=NC2=C(C(N1CC3=CC=CO3)=O)NC(C)=C2)NCC4=CC=C(CC)C=C4
Synonyms

ZINC67299692
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Handling Instructions (2659 KB)

References

[1]. Kadivella M, et al. Adjuvant activity of a small molecule TLR4 agonist discovered via structure-based virtual screening. Communications biology. 2025 Sep 29;8(1):1382.

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Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

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Biological Activity
- Description
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References

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NSF-951

All Toll-like Receptor (TLR) Isoforms

All p38 MAPK Isoforms

All JNK Isoforms

All Interleukin Related Isoforms

All TNF Receptor Isoforms