RESUMO
Protein misfolding into amyloid fibrils is linked to more than 40 as yet incurable cell- and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and typeâ 2 diabetes. So far, however, only one of the numerous anti-amyloid molecules has reached patients. This Minireview gives an overview of molecular strategies and peptide chemistry "tools" to design, develop, and discover peptide-based molecules as anti-amyloid drug candidates. We focus on two major inhibitor rational design strategies: 1)â the oldest and most common strategy, based on molecular recognition elements of amyloid self-assembly, and 2)â a more recent approach, based on cross-amyloid interactions. We discuss why peptide-based amyloid inhibitors, in particular their advanced generations, can be promising leads or candidates for anti-amyloid drugs as well as valuable tools for deciphering amyloid-mediated cell damage and its link to disease pathogenesis.
Assuntos
Amiloide/antagonistas & inibidores , Peptídeos/química , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Amiloide/metabolismo , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Desenho de Fármacos , Humanos , Peptídeos/metabolismo , Peptídeos/farmacologia , Agregados Proteicos/efeitos dos fármacos , Dobramento de ProteínaRESUMO
The interaction of the intrinsically disordered polypeptide islet amyloid polypeptide (IAPP), which is associated with type 2 diabetes (T2D), with the Alzheimer's disease amyloid-ß (Aß) peptide modulates their self-assembly into amyloid fibrils and may link the pathogeneses of these two cell-degenerative diseases. However, the molecular determinants of this interaction remain elusive. Using a systematic alanine scan approach, fluorescence spectroscopy, and other biophysical methods, including heterocomplex pulldown assays, far-UV CD spectroscopy, the thioflavin T binding assay, transmission EM, and molecular dynamics simulations, here we identified single aromatic/hydrophobic residues within the amyloid core IAPP region as hot spots or key residues of its cross-interaction with Aß40(42) peptide. Importantly, we also find that none of these residues in isolation plays a key role in IAPP self-assembly, whereas simultaneous substitution of four aromatic/hydrophobic residues with Ala dramatically impairs both IAPP self-assembly and hetero-assembly with Aß40(42). Furthermore, our experiments yielded several novel IAPP analogs, whose sequences are highly similar to that of IAPP but have distinct amyloid self- or cross-interaction potentials. The identified similarities and major differences controlling IAPP cross-peptide interaction with Aß40(42) versus its amyloid self-assembly offer a molecular basis for understanding the underlying mechanisms. We propose that these insights will aid in designing intervention strategies and novel IAPP analogs for the management of type 2 diabetes, Alzheimer's disease, or other diseases related to IAPP dysfunction or cross-amyloid interactions.
Assuntos
Aminoácidos/metabolismo , Peptídeos beta-Amiloides/metabolismo , Polipeptídeo Amiloide das Ilhotas Pancreáticas/metabolismo , Modelos Moleculares , Fragmentos de Peptídeos/metabolismo , Agregação Patológica de Proteínas/metabolismo , Substituição de Aminoácidos , Aminoácidos/química , Aminoácidos Aromáticos , Peptídeos beta-Amiloides/síntese química , Peptídeos beta-Amiloides/química , Dicroísmo Circular , Humanos , Interações Hidrofóbicas e Hidrofílicas , Polipeptídeo Amiloide das Ilhotas Pancreáticas/síntese química , Polipeptídeo Amiloide das Ilhotas Pancreáticas/química , Cinética , Metilação , Microscopia Eletrônica de Transmissão , Simulação de Dinâmica Molecular , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/química , Estabilidade Proteica , Estrutura Secundária de Proteína , Técnicas de Síntese em Fase Sólida , Solubilidade , Espectrometria de FluorescênciaRESUMO
Amyloid self-assembly is linked to the pathogenesis of Alzheimer's disease (AD) and typeâ 2 diabetes (T2D), but so far, no anti-amyloid compound has reached the clinic. Macrocyclic peptides belong to the most attractive drug candidates. Herein we present macrocyclic peptides (MCIPs) designed using minimal IAPP-derived recognition elements as a novel class of nanomolar amyloid inhibitors of both Aß40(42) and IAPP or Aß40(42) alone and show that chirality controls inhibitor selectivity. Sequence optimization led to the discovery of an Aß40(42)-selective MCIP exhibiting high proteolytic stability in human plasma and human blood-brain barrier (BBB) crossing ability in a cell model, two highly desirable properties for anti-amyloid AD drugs. Owing to their favorable properties, MCIPs should serve as leads for macrocyclic peptide-based anti-amyloid drugs and scaffolds for the design of small-molecule peptidomimetics for targeting amyloidogenesis in AD or in both AD and T2D.
Assuntos
Amiloide/antagonistas & inibidores , Compostos Macrocíclicos/química , Compostos Macrocíclicos/farmacologia , Nanotecnologia , Peptídeos/química , Peptídeos/farmacologia , Sequência de AminoácidosRESUMO
The design of inhibitors of protein-protein interactions mediating amyloid self-assembly is a major challenge mainly due to the dynamic nature of the involved structures and interfaces. Interactions of amyloidogenic polypeptides with other proteins are important modulators of self-assembly. Here we present a hot-segment-linking approach to design a series of mimics of the IAPP cross-amyloid interaction surface with Aß (ISMs) as nanomolar inhibitors of amyloidogenesis and cytotoxicity of Aß, IAPP, or both polypeptides. The nature of the linker determines ISM structure and inhibitory function including both potency and target selectivity. Importantly, ISMs effectively suppress both self- and cross-seeded IAPP self-assembly. Our results provide a novel class of highly potent peptide leads for targeting protein aggregation in Alzheimer's disease, type 2 diabetes, or both diseases and a chemical approach to inhibit amyloid self-assembly and pathogenic interactions of other proteins as well.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Desenho de Fármacos , Polipeptídeo Amiloide das Ilhotas Pancreáticas/farmacologia , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/química , Peptídeos beta-Amiloides/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Polipeptídeo Amiloide das Ilhotas Pancreáticas/química , Agregados Proteicos/efeitos dos fármacos , Propriedades de SuperfícieRESUMO
Growth differentiation factor 15 (GDF15), a member of the transforming growth factor ß superfamily, has been postulated to be implicated in cancer cell metastasis although its role has not been fully elucidated yet. The purpose of this review is to clarify the role of GDF-15 in cancer cell metastasis based on current advances in the field. The studies were divided into those involving evaluation of GDF15 expression in the serum or tissue of cancer patients, and those involving in vitro experiments in cancer cell lines or in vivo experiments in animal models. GDF15 was shown to be elevated in the serum or tissues of cancer patients with its expression being correlated with decreased survival. Moreover, most in vitro and in vivo studies also corroborated a metastasis-promoting role for GDF15. However, there were a few studies, where GDF15 was shown to suppress the metastatic properties of cells. As, GDF15 has been known for its pleiotropic effects, it is not surprising to behave differently in different types of cancer. Thus, GDF15 has the potential of not only being a useful metastasis biomarker, but also a promising therapeutic target against cancer cell metastasis in many cancer types.
Assuntos
Fator 15 de Diferenciação de Crescimento/metabolismo , Metástase Neoplásica/patologia , Neoplasias/patologia , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Invasividade Neoplásica/patologiaRESUMO
Polysorbates (PSs) are the most common surfactants in therapeutic protein formulations, and it is crucial to monitor their concentration along the life cycle of biopharmaceuticals. We developed a simple multi-well plate fluorescence-based assay for the rapid determination of PS20 and PS80 content in biopharmaceutical products. The method is based on the detection of the fluorescence emission intensity of the fluorescent dye 1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate in the presence of PSs at concentrations below their critical micelle concentration. This method can be applied for PS content determination in protein formulations (≤100 mg/mL) without the need of a previous protein removal step. The 1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate assay implemented in multi-well plate format is suitable for high-throughput concentration screening. It has a linear range from 0.00020% to 0.0025% (w/v) PS20, and the limits of detection and quantification were 0.00020% and 0.00055% (w/v), respectively. This assay is markedly more selective and shows no or lower interferences due to hydrophobic components (e.g., silicone oil) potentially present in finished products than the fluorescence micelle assay based on N-phenyl-1-naphthylamine. It also provides comparable results for the PS content in liquid chromatography with charged aerosol detection analysis with protein removal, providing a fast alternative.