Antihypertensive active peptide Orn-Pro-Hyp and application and medicinal composition
An anti-hypertensive and active peptide technology, applied in the field of biomedicine, can solve problems such as low repeatability, constraints, and high costs, and achieve the effect of ensuring integrity
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Embodiment 1
[0019] Example 1: Synthesis of Orn-Pro-Hyp tripeptide.
[0020] The Orn-Pro-Hyp tripeptide used in the present invention can be synthesized by artificial chemical methods known in the field of polypeptide synthesis. In this embodiment, a liquid-phase synthesis method is used, and the specific operations are as follows:
[0021] First, choose the preloaded tert-butyl hydroxyproline dichlorotrityl resin (Hyp(Tbu)-2Cl(TRT)-resin) with O-benzotriazole-N,N,N', N'-Tetramethylurea tetrafluoroboric acid (TBTU) and N,N-diisopropylethylamine (DIEA) are used as condensation agents, adding fluorenylmethoxycarbonyl-L-proline (FMOC-Pro-OH), React for 1 hour. Next, use dimethylformamide (DMF) as a cleaning agent to clean the product obtained in the previous step reaction to obtain fluorenylmethoxycarbonyl prolyl tert-butyl hydroxyproline dichlorotrityl resin (FMOC-Pro- Hyp(Tbu)-2Cl(TRT)-resin). Then, adopt hexahydropyridine (PPD) as deprotecting agent, join in the resin (FMOC-Pro-Hyp(Tbu)...
Embodiment 2
[0029] Example 2: In vitro Orn-Pro-Hyp inhibition test on angiotensin-converting enzyme (ACE) activity.
[0030] 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 0.76mU / 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, It is used to evaluate the inhibition rate of ACE enzyme inhib...
Embodiment 3
[0035] Example 3: In vivo antihypertensive test in congenitally hypertensive rats (SHR).
[0036] 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.
[0037] 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 blood pressure of the rats before gavage, then 30mg / kg body weight dose is gavaged with the sample (active tripeptide, Orn-Pro-Hyp), the blank control group is gavaged with an equal volume of normal saline (Saline), and the drug control gro...
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