Application of dipeptidyl peptidase IV (DPP4) inhibitors to preparation of drugs used for preventing and treating epilepsy
A dipeptidyl peptidase and inhibitor technology, which is applied in the field of medical applications to achieve the effects of less toxic and side effects, extremely high toxic and side effects, and long duration of drug efficacy
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Embodiment 1
[0033] [Example 1] Studying the effect of DPP4 inhibitors on the behavior of epileptic rats
[0034] Select 30 (220±20g) male SD rats and randomly divide them into 3 groups: control group, epilepsy group and sitagliptin phosphate treatment group. Control group: intraperitoneal injection of normal saline at 35 mg / kg; epilepsy group: intraperitoneal injection of PTZ at 35 mg / kg to construct an epilepsy model; treatment group: intraperitoneal injection of sitagliptin phosphate at 10 mg / kg 20 minutes in advance, and 35 mg / kg 20 minutes later The epilepsy model was established by intraperitoneal injection of PTZ at kg. According to the Racine grading standard (1972), the grades of epileptic seizures are graded from 0 to V. The main manifestations are: grade 0 is normal (no convulsions); Grade II rhythmic nodding and chewing; Grade III unilateral forelimb spasm; Grade IV generalized tonic-clonic convulsions with standing, falling, and retreating; Grade V recurrent generalized tonic...
Embodiment 2
[0036] [Example 2] Studying the effects of DPP4 inhibitors on neuronal discharge in epileptic rats
[0037] In a quiet environment at room temperature, SD rats were injected intraperitoneally with 10% chloral hydrate (0.3mL / 100g) in a state of general anesthesia, and a small hand drill with a diameter of 0.5mm was used to place them 4mm behind the anterior bregma and 4mm on both sides of the midline. Two recording electrode holes were drilled in the skull of the rat, and one reference electrode hole was drilled above the sinuses of the rat. Two insulated stainless steel electrodes were respectively placed in the reference electrode hole and the recording electrode hole on the right, and then all the electrodes were fixed on the rat skull with dental cement (pay attention to aseptic operation during the above operation). Connect the electrode leads to the digital EEG machine, adjust the EEG parameters, and record the discharge of neurons in each group of rats.
[0038] The exp...
Embodiment 3
[0039] [Example 3] Studying the Effect of DPP4 Inhibitors on the Survival Rate of Neurons in Epilepsy Rats
[0040] The main steps are as follows:
[0041] (1) Tissue section: 24 hours after the epileptic behavior observation of SD rats, 10% chloral hydrate (0.3mL / 100g) was injected intraperitoneally to make the rats under general anesthesia, and the left ventricle was perfused and fixed with 4% paraformaldehyde. During the perfusion process, the muscles of the whole body of the rat can be observed to vibrate, and gradually become stiff and rigid. At this time, the brain is decapitated, and then the tissue is paraffin-embedded and sectioned for later use.
[0042](2) Nissl staining: wash the sections sequentially in xylene I 15 min-xylene II 15 min absolute ethanol I 5 min-absolute ethanol II 5 min-85% alcohol 5 min-75% alcohol 5 min-distilled water. The paraffin sections were stained in buffered methylene blue staining solution for 10 min. Differentiate in 0.2 mol / L acetate...
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