Background and overview[1][2]
Nicotinamide adenine dinucleotide phosphate is also called coenzyme II, a type of dehydrogenase coenzyme. It is the condensate of nicotinamide riboside-3′,5′-diphosphate and adenine nucleotide. There are two forms: oxidation type and reduction type, abbreviated as NADP+ flame-resistant hydraulic oil and NADPH+H+. Contains vitamin B5, which has the same mechanism of action in redox reactions as nicotinamide adenine dinucleotide. It is a coenzyme of some non-aerobic dehydrogenases and plays a role in transmitting hydrogen in organisms. Nicotinamide adenine dinucleotide phosphate (NADP) is a coenzyme that widely exists in the biological world and participates in many physiological processes; research shows that the nicotinamide adenine dinucleotide phosphate NADP It is mainly reduced to NADPH in cells, which is an important antioxidant component in cells and also participates in the anabolic process.
Indications[2]
Basic and clinical research in recent years has confirmed that oxidative stress damage is one of the main pathogenesis mechanisms of diabetic nephropathy. Transgenic technology is used to overexpress antioxidant enzymes in the kidney, such as superoxide dismutase (SOD) or hydrogen peroxide. Enzyme (catalase) can significantly improve diabetic kidney damage. Based on the research foundation and status of existing technology, CN201610070262.4 provides new medicinal uses of the compound nicotinamide adenine dinucleotide phosphate (NADP), specifically involving the compound nicotinamide adenine dinucleotide phosphate Use of NADP in preparing drugs for preventing and treating diabetic nephropathy.
CN201610070262.4 has confirmed through in vitro cell experiments and in vivo animal experiments that the NADP can reduce the damage of high sugar to vascular endothelial cells, improve kidney damage in diabetic mice, and can further be used to prepare drugs for preventing and treating diabetic nephropathy. The medicine prepared according to the present invention can be used by intraperitoneal injection. The dose of NADP used is 5-20mg.kg-1.d-1. Start using it. The optimal time is when diabetes is diagnosed, which can prevent and treat diabetic nephropathy. The present invention provides new targets and ideas for clinical prevention and treatment of diabetic nephropathy and improvement of renal lesions, and confirms the important role of improving kidney oxidative stress in the prevention and treatment of diabetic nephropathy.
Apply[3]
Creatine kinase isoenzyme assay kit (selective inhibition method) is used to determine the content of creatine kinase isoenzyme in human serum or plasma. The most important significance of serum CK-MB (creatine kinase isoenzyme) determination is in the diagnosis of acute myocardial infarction. Clinically, CK-MB exceeding the total activity of CK is often used as the basis for the diagnosis of acute myocardial infarction. Creatine kinase is a kinase that plays a vital role in intracellular energy transport and metabolism. Creatine kinase isoenzyme is an isoenzyme of creatine kinase, which is generated after post-translational modification. It mainly exists in cardiomyocytes (and a small amount of it exists in bone muscles, intestines, tongue, prostate and In utero), it is a very important creatine kinase in the myocardial energy metabolism pathway. It is composed of two subunits, representing muscle and brain.
The principle of testing creatine kinase isoenzymes is that the main isoenzymes of creatine kinase (CK) include CKMB, CKBB, and CKMM. Because there is almost no CKBB in normal serum, the antiserum against the M subunit can be combined with CKMM and CKMM. The M subunit in CKMB forms an antigen-antibody complex, thereby completely inhibiting the enzyme activity of the M subunit. Multiplying the measured CK activity value by 2 represents the activity of CKMB. The detection principle of creatine kinase (CK) activity is: CK catalyzes the reaction of creatine phosphate and adenosine diphosphate (ADP) to produce adenosine triphosphate (ATP) and creatine; coupled hexokinase (HK) and glucose-6-phosphate detoxification Catalytic reaction of hydrogenase (G6PDH). HK catalyzes the reaction between glucose and ATP to form glucose-6-phosphate, and G6PDH catalyzes the oxidation of glucose-6-phosphate to form 6-phosphate glucose lactone and NADPH. The rate of NADPH generation represents CK activity.
�Since the antigen is a protein, its stability is extremely poor and will be completely degraded within hours at room temperature. After the creatine kinase isoenzyme meets the clinical requirements, its stability will be evaluated. One of the difficulties to be overcome in the Creatine Kinase Isoenzyme Determination Kit (Immunosuppression Method) is the stability of the reagent, which must meet the requirements of a deviation of less than 10% for each performance index after 14 days of heat breakage at 7°C. However, it is difficult for existing reagents to meet the above requirements.
CN201711008283.4 provides a creatine kinase isoenzyme dual reagent and a preparation method thereof. When stored at 37°C for 14 days, the present invention still maintains high reagent stability, and the measurement value deviation of the reagent is less than 10%. The method includes the following steps:
1) Weigh N-acetylcysteine, creatine phosphate, adenosine diphosphate, adenosine triphosphate, glucose-6-phosphate dehydrogenase, hexokinase, creatine kinase isoenzyme antibody, and polyvinylpyrrolidone , lauryl dihydroxyethyl amine oxide and sodium isomeric ascorbate, dissolve in water to obtain a mixed solution, mix the mixed solution with imidazole buffer, add tris(nonylphenyl) phosphite to dissolve, and obtain reagent I Solution;
2) Weigh nicotinamide adenine dinucleotide phosphate and glucose and dissolve them in water to obtain a reagent II solution.
The beneficial effects of the present invention are: polyvinylpyrrolidone, lauryldihydroxyethylamine oxide and tris(nonylphenyl)phosphite are added to the creatine kinase isoenzyme assay kit reagent I of the present invention. In addition, It also contains the antioxidant sodium isomerized ascorbate, which improves the storage stability of the reagent. It has been verified that the shelf life of the present invention can be as long as one and a half years at 2-8°C, which greatly improves the actual shelf life and its transportation stability. It is of great help to rural hospitals in remote mountainous areas with small sample sizes. And when stored at 37°C for 14 days, the present invention still maintains high reagent stability, and the deviation of the measured value of the reagent is less than 10%.
Main reference materials
[1] Agricultural Dictionary
[2] CN201610070262.4 Use of nicotinamide adenine dinucleotide phosphate in the preparation of drugs for preventing and treating diabetic nephropathy
[3] CN201711008283.4 A creatine kinase isozyme dual reagent and its preparation method
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