Antihypertensive bioactive peptide Citn-Hyp-Pro as well as application and medicine composition
A citn-hyp-pro, anti-hypertensive technology, applied in the field of biomedicine, can solve the problems of low repeatability, high cost and low bioavailability
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Embodiment 1
[0020] Synthesis of Citn-Hyp-Pro tripeptide. The antihypertensive active peptide of the present invention is artificially chemically synthesized, and the specific operation is as follows:
[0021] The polypeptide of the present invention is synthesized by a liquid-phase polypeptide synthesis method. A certain amount of N-tert-butoxycarbonyl citrulline methyl ester (Boc-Cit) and hydroxysuccinimide (Hosu) are placed in a round-bottomed flask, and tetrahydrofuran is added. (THF) was dissolved until it became clear, and dicyclohexylcarbodiimide (DCC) was continuously added, stirred gently, and reacted overnight.
[0022] Further, the above reaction solution was vacuum filtered, and ethyl acrylate (EA) and H 2 O for chromatography. Absorb the upper liquid, using NaHCO 3 / H 2 Wash twice with O, then wash with citric acid / H 2 Washed twice with O, washed twice with saturated NaCl solution, Na 2 SO 4 Dry and evaporate to dryness with a rotary evaporator to obtain N-tert-butoxyca...
Embodiment 2
[0030] Angiotensin-converting enzyme (ACE) activity inhibition test in vitro.
[0031]Hippuryl-L-histidyl-L-leucine (hippuryl-L-histidyl-L-leucine, HHL) is rapidly decomposed under the catalysis of ACE enzyme to produce hippuric acid (Hippuric Acid, HA) and dipeptide histidyl-leucine (HL), adding After ACE enzyme inhibitor, the activity of ACE enzyme is inhibited, and the production of HA and HL is reduced. In this embodiment, ACE is extracted from rabbit lung, and the enzyme activity is 3.19mU / mL. It is developed by DAB and measured by spectrophotometer. The amount of HA produced and the activity of ACE enzyme were analyzed. Ethyl acetate extracts hippuric acid in the reactant, then reacts with a pyridine solution (DAB chromogen) containing p-dimethylaminobenzaldehyde in acetic anhydride to generate an orange-yellow compound, and directly measures its OD value at 459nm colorimetrically, The inhibition rate of ACE enzyme inhibitors to ACE enzyme was evaluated according to the...
Embodiment 3
[0038] In vivo antihypertensive test in congenitally hypertensive rats (SHR).
[0039] Using SoftronBP-98A intelligent non-invasive blood pressure instrument for rats, the systolic blood pressure (SBP) of the rats was determined by the tail method.
[0040] In the awake state of SHR rats (spontaneously hypertensive rats), first put the rats in a rat bag, keep a constant temperature, and use the Softron BP-98A rat intelligent non-invasive blood pressure instrument to measure the tail vein blood pressure. Systolic blood pressure (SBP) in rats. One week before the experiment, the blood pressure of the rats was measured every other day, and the experimental records were started after the rats got used to it stably. First measure the rat blood pressure before gavage, then 1.0mg / kg body weight dose carries out sample (active tripeptide of embodiment 1, Citn-Hyp-Pro) gavage, blank control group gavage equal volume normal saline (Saline), The drug control group was administered intr...
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