2-Chloro-4-nitrophenyl a-D-fucopyranoside

2-Chloro-4-nitrophenyl a-D-fucopyranoside (CNPF) is a chromogenic enzyme substrate used primarily to detect fucosidase activity. CNPF is cleaved by the enzyme, which results in the release of a colored product that can be measured spectrophotometrically. Fucosidases are important enzymes that catalyze the hydrolysis of fucose-containing oligosaccharides, and CNPF is a commonly used substrate for the detection and quantification of these enzymes in various research studies.

2-Chloro-4-nitrophenyl α-D-fucopyranoside (CNPF) is a synthetic substrate widely used in enzymatic assays to detect and quantify glycosidases activity. It is a small molecule that exhibits favorable physicochemical and biological properties, making it an excellent tool for various molecular biology applications.

Physical and Chemical Properties:

CNPF is a white crystalline powder with a molecular weight of 329.64 g/mol. It is highly soluble in water, methanol, and ethanol but insoluble in benzene and chloroform. CNPF's melting point is 231-233°C, and it has a strong ultraviolet (UV) absorption maximum at 364 nm.

Synthesis and Characterization:

CNPF is synthesized by the reaction of p-nitrophenyl α-L-fucopyranoside and thionyl chloride in chloroform, followed by chlorination with oxalyl chloride.

The characterization of CNPF involves different techniques, including nuclear magnetic resonance (NMR) spectroscopy, high-performance liquid chromatography (HPLC), and mass spectrometry (MS).

Analytical Methods:

Different analytical methods have been used to determine CNPF concentrations, including HPLC, UV-visible spectrophotometry, and capillary electrophoresis. These techniques rely on the absorption or fluorescence of CNPF molecules or the enzymatic cleavage of CNPF by glycosidases, which can be monitored by various means.

Biological Properties:

As a glycosidase substrate, CNPF is hydrolyzed by a wide range of glycosidases, including α-L-fucosidase, β-galactosidase, and β-glucosidase. CNPF has been used to detect the presence and activity of glycosidases in different organisms, including bacteria, fungi, plants, and animals.

Toxicity and Safety in Scientific Experiments:

CNPF is generally considered safe and non-toxic when used in scientific experiments. However, it is important to handle it with care, as it can be harmful if ingested, inhaled, or comes in contact with skin or eyes.

Applications in Scientific Experiments:

CNPF has numerous applications in scientific experiments, including:

- Detection and quantification of glycosidase activity

- Screening for glycosidase inhibitors

- Structural and functional studies of glycosidases

- Development of diagnostic assays for glycosidase-related diseases.

Current State of Research:

CNPF is widely used in molecular biology, biochemistry, and drug discovery research. Recent studies have focused on developing more efficient, sensitive, and specific methods for glycosidase detection and inhibition using CNPF. Also, different strategies have been developed to enhance the biocompatibility and targeting of CNPF-based probes and inhibitors.

Potential Implications in Various Fields of Research and Industry:

CNPF has potential implications in various fields of research and industry, including:

- Development of glycosidase-based diagnostic and therapeutic tools for glycosidase-related diseases, including cancer, genetic disorders, and viral infections.

- Development of glycosidase inhibitors as potential drugs for the treatment of metabolic disorders, cardiovascular diseases, and inflammation.

- Use of CNPF-based probes for imaging and targeting of glycosidase activity in vivo and in vitro.

Limitations and Future Directions:

Despite its many advantages, CNPF has some limitations in its applications, including its susceptibility to air and light degradation, low solubility in organic solvents, and limited specificity towards certain glycosidases.

Future directions for CNPF research include:

- Development of more stable and specific CNPF derivatives and analogs

- Optimization of CNPF-based assays and inhibitors for high-throughput screening and clinical applications

- Investigation of the role of glycosidases and their inhibitors in different physiological and pathological processes

- Development of novel glycosidase-targeted therapies and imaging techniques.