Application of nicotinamide phosphoribosyltransferase in preparation of neuroprotective drugs

Abstract

Belonging to the pharmaceutical field, the invention relates to application of nicotinamide phosphoribosyltransferase (NAMPT) in preparation of neuroprotective and rehabilitation drugs. Experiments of the invention show that NAMPT recombinant protein can penetrate the blood-brain barrier to significantly reduce the volume of ischemic brain injury and alleviate white matter injury after ischemic brain injury; the NAMPT recombinant protein can significantly improve long-term behaviors after ischemic brain injury; the NAMPT recombinant protein can raise revascularization related factors to enhance revascularization after brain hypoxia and neuron regeneration after brain hypoxia, and promote post-injury neurologic function reestablishment; and by mediating oligodendrocyte regeneration and alleviating the demyelination effect of local lysolecithin, the NAMPT recombinant protein can promote restoration of white matter after injury and strengthen post-injury neurologic function reestablishment. Results show that the NAMPT recombinant protein plays a long-term neuroprotective role in drugs treating cerebral stroke, traumatic brain injury and multiple sclerosis, and can be used as a neuroprotective drug to treat acute and chronic injury caused by brain stroke.

Description

Technical field:

[0001] The invention belongs to the field of pharmacy and relates to a new medical application of nicotinamide phosphoribosyltransferase (NAMPT). It specifically relates to the use of NAMPT nicotinamide phosphoribosyltransferase in the preparation of neuroprotective and rehabilitation medicines. Especially in the preparation of medicines for the treatment of central nervous system diseases. Background technique

[0002] The China Stroke Conference organized by the Ministry of Health of China in 2012 pointed out that stroke, especially ischemic stroke, has become the disease with the highest incidence in my country. So far, the only clinically effective therapeutic drug is thrombolytic therapy (tPA), but practice shows that the application of tPA is very limited, because tPA has a strict therapeutic window, and it is only effective if intravenous injection within 3 hours after stroke. Therefore, less than 3% of patients can be treated with tPA clinically. ...

Examples

Embodiment 1

[0044] 1. Experimental materials and methods

[0045] 1.1 Synthesis of NAMPT recombinant protein

[0046] Since glycosylation is the necessary way for NAMPT protein secretion, the present invention uses eukaryotic expression plasmids to express NAMPT protein, and uses dihydrofolate dehydrogenase-methotrexate system to selectively amplify in CHO cell lines . In order to increase the concentration of NAMPT protein in the culture medium, in this embodiment, the secretion signal peptide of the Ig Kappa chain is connected to the N-terminus of NAMPT, and the His6 tag is connected to the C-terminus of NAMPT for purification and separation. The synthetic cDNA is inserted into the cDNA containing Dihydrofolate dehydrogenase expression plasmid system (plgK-NAMPT-His6), and finally this vector was transfected into Chinese hamster cell line DG44 (DHFR- / -) without dihydrofolate dehydrogenase, using α-MEM medium +10% nucleoside-free fetal bovine serum for culture, adding methotrexate (5Nm...

Technical field:

[0001] The invention belongs to the field of pharmacy and relates to a new medical application of nicotinamide phosphoribosyltransferase (NAMPT). It specifically relates to the use of NAMPT nicotinamide phosphoribosyltransferase in the preparation of neuroprotective and rehabilitation medicines. Especially in the preparation of medicines for the treatment of central nervous system diseases. Background technique

[0002] The China Stroke Conference organized by the Ministry of Health of China in 2012 pointed out that stroke, especially ischemic stroke, has become the disease with the highest incidence in my country. So far, the only clinically effective therapeutic drug is thrombolytic therapy (tPA), but practice shows that the application of tPA is very limited, because tPA has a strict therapeutic window, and it is only effective if intravenous injection within 3 hours after stroke. Therefore, less than 3% of patients can be treated with tPA clinically. ...

Examples

Embodiment 1

[0044] 1. Experimental materials and methods

[0045] 1.1 Synthesis of NAMPT recombinant protein

[0046] Since glycosylation is the necessary way for NAMPT protein secretion, the present invention uses eukaryotic expression plasmids to express NAMPT protein, and uses dihydrofolate dehydrogenase-methotrexate system to selectively amplify in CHO cell lines . In order to increase the concentration of NAMPT protein in the culture medium, in this embodiment, the secretion signal peptide of the Ig Kappa chain is connected to the N-terminus of NAMPT, and the His6 tag is connected to the C-terminus of NAMPT for purification and separation. The synthetic cDNA is inserted into the cDNA containing Dihydrofolate dehydrogenase expression plasmid system (plgK-NAMPT-His6), and finally this vector was transfected into Chinese hamster cell line DG44 (DHFR- / -) without dihydrofolate dehydrogenase, using α-MEM medium +10% nucleoside-free fetal bovine serum for culture, adding methotrexate (5Nm...

Images