An orally active formulation of angiotensin-(1-7) produces an antithrombotic effect

INTRODUCTION AND OBJECTIVE: The heptapeptide angiotensin-(1-7) is a component of the renin-angiotensin system, which promotes many beneficial cardiovascular effects, including antithrombotic activity. We have recently shown that the antithrombotic effect of angiotensin-(1-7) involves receptor Mas-mediated NO-release from platelets. Here, we describe an orally active formulation based on angiotensin-(1-7) inclusion in cyclodextrin [Ang-(1-7)- CyD] as an antithrombotic agent. Cyclodextrins are pharmaceutical tools that are used to enhance drug stability, absorption across biological barriers and gastric protection. METHOD: To test the antithrombotic effect of Ang-(1-7)-CyD, thrombus formation was induced in the abdominal vena cava of spontaneously hypertensive rats that were pretreated either acutely or chronically with Ang-(1-7)-CyD. Male Mas-knockout and wild-type mice were used to verify the role of the Mas receptor on the effect of Ang-(1-7)-CyD. RESULTS: Acute or chronic oral treatment with Ang-(1-7)-CyD promoted an antithrombotic effect (measured by thrombus weight; all values are, respectively, untreated vs. treated animals) in spontaneously hypertensive rats (acute: 2.86 + 0.43 mg vs. 1.14 + 0.40 mg; chronic: 4.27 + 1.03 mg vs. 1.39 + 0.68 mg). This effect was abolished in Mas-knockout mice (thrombus weight in Mas wild-type: 0.76 + 0.10 mg vs. 0.37 + 0.02 mg; thrombus weight in Mas-knockout: 0.96 + 0.11 mg vs. 0.87 + 0.14 mg). Furthermore, the antithrombotic effect of Ang-(1-7)-CyD was associated with an increase in the plasma level of Angiotensin-(1-7). CONCLUSION: These results show for the first time that the oral formulation Ang-(1-7)-CyD has biological activity and produces a Mas-dependent antithrombotic effect.

Study of the BPP7a Peptide and its Beta Cyclodextrin Complex: PhysicochemicalCharacterization and Complete Sequence Specific NMR Assignments

O heptapeptídeo BPP7a, p-Glu1Asp2Gly3Pro4Ile5Pro6Pro7, forma um complexo de associaçãocom a β-ciclodextrina na razão molar 1:1. O peptídeo e a sua forma complexa foram caracterizados por dicroísmo circular (CD) e titulação calorimétrica (ITC), as quais sugerem uma interação muito fraca entre a β-ciclodextrina e o peptídeo. Espectros de ressonância magnética nuclear (NMR) de 1H a 400 e 600 MHz foram obtidos para o peptídeo puro e para o complexo com β-ciclodextrina e com estes foi possível a atribuição de todos os sinais de ressonância de hidrogênio do peptídeo. Experimentos de espectroscopia ordenada de difusão de alta resolução (HR-DOSY) foram conduzidos a fim de se confirmar a associação entre o BPP7a e a β-ciclodextrina, além de se verificar a quebra dos agregados moleculares do BPP7a devida a associação. A atividade antihipertensiva do complexo BPP7a/β-ciclodextrina foi avaliada em ratos naturalmente hipertensivos (SHR), mostrando resultados melhores do que os do peptídeo BPP7a puro.

The BPP7a heptapeptide, p-Glu1Asp2Gly3Pro4Ile5Pro6Pro7, forms an association complexwith β-cyclodextrin in a 1:1 molar ratio. The peptide and its complex were characterized bycircular dichroism (CD) and isothermal titration calorimetry (ITC), which showed a very weak interaction between the β-cyclodextrin and the peptide. Assignments of all hydrogen resonances of the peptide alone and as a complex were made using 1H nuclear magnetic resonance (NMR)experiments at 400 and 600 MHz. High resolution diffusion ordered spectroscopy (HR-DOSY) experiments were carried out to establish the self-aggregation state of BPP7a. It was also shown that the β-cyclodextrin breaks the molecular clusters leading to complex formation. In addition,the anti-hypertensive activity of the BPP7a/β-cyclodextrin complex was evaluated in spontaneous hypertensive rats (SHR), showing increased activity compared to that of pure BPP7a.