RESUMO
Purinergic receptor is a potential drug target for neuropathic pain, Alzheimer disease, and prostate cancer. Focusing on the structure-based ligand discovery, docking analysis on the crystal structure of P2Y1 receptor (P2Y1R) with 923 derivatives of 1-indolinoalkyl 2-phenolic compound is performed to understand the molecular insights of the receptor. The structural model identified the top novel ligands, 426 (compound 1) and 636 (compound 2) having highest binding affinity with the docking score of -7.38 and -6.92. We have reported the interaction efficacy and the dynamics of P2Y1R protein with the ligands. The best hits synthesized were experimentally optimized as a potent P2Y1 agonists. These ligands exhibits anti-proliferative effect against the PC-3 and DU-145 cells (IC50 = 15 µM - 33 µM) with significant increase in the calcium level in dose- and time-dependent manner. Moreover, the activation of P2Y1R induced the apoptosis via Capase3/7 and ROS signaling pathway. Thus it is evidenced that the newly synthesized ligands, as a P2Y1R agonists could potentially act as a therapeutic drug for treating prostate cancer.
Assuntos
Antineoplásicos , Simulação de Acoplamento Molecular , Proteínas de Neoplasias , Neoplasias da Próstata , Agonistas do Receptor Purinérgico P2Y , Receptores Purinérgicos P2Y1 , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Células HEK293 , Humanos , Ligantes , Masculino , Camundongos , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Células PC-3 , Neoplasias da Próstata/química , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Agonistas do Receptor Purinérgico P2Y/síntese química , Agonistas do Receptor Purinérgico P2Y/química , Agonistas do Receptor Purinérgico P2Y/farmacologia , Receptores Purinérgicos P2Y1/química , Receptores Purinérgicos P2Y1/metabolismoRESUMO
Dinucleoside 5',5'-polyphosphates (DNPs) are endogenous substances that play important intra- and extracellular roles in various biological processes, such as cell proliferation, regulation of enzymes, neurotransmission, platelet disaggregation and modulation of vascular tone. Various methodologies have been developed over the past fifty years to access these compounds, involving enzymatic processes or chemical procedures based either on P(III) or P(V) chemistry. Both solution-phase and solid-support strategies have been developed and are reported here. Recently, green chemistry approaches have emerged, offering attracting alternatives. This review outlines the main synthetic pathways for the preparation of dinucleoside 5',5'-polyphosphates, focusing on pharmacologically relevant compounds, and highlighting recent advances.
Assuntos
Fosfatos de Dinucleosídeos/síntese química , Agonistas do Receptor Purinérgico P2Y/síntese química , Nucleotídeos de Desoxicitosina/agonistas , Nucleotídeos de Desoxicitosina/química , Nucleotídeos de Desoxicitosina/farmacologia , Fosfatos de Dinucleosídeos/química , Fosfatos de Dinucleosídeos/isolamento & purificação , Síndromes do Olho Seco/tratamento farmacológico , Química Verde , Humanos , Soluções Oftálmicas , Fosforilação , Polifosfatos/síntese química , Polifosfatos/química , Agonistas do Receptor Purinérgico P2Y/química , Agonistas do Receptor Purinérgico P2Y/isolamento & purificação , Receptores Purinérgicos/metabolismo , Nucleotídeos de Uracila/química , Uridina/agonistas , Uridina/análogos & derivados , Uridina/química , Uridina/farmacologiaRESUMO
Aberrant nucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) activity is associated with chondrocalcinosis, osteoarthritis, and type 2 diabetes. The potential of NPP1 inhibitors as therapeutic agents, and the scarceness of their structure-activity relationship, encouraged us to develop new NPP1 inhibitors. Specifically, we synthesized ATP-α-thio-ß,γ-CH2 (1), ATP-α-thio-ß,γ-CCl2 (2), ATP-α-CH2-γ-thio (3), and 8-SH-ATP (4) and established their resistance to hydrolysis by NPP1,3 and NTPDase1,2,3,8 (<5% hydrolysis) (NTPDase = ectonucleoside triphosphate diphosphohydrolase). Analogues 1-3 at 100 µM inhibited thymidine 5'-monophosphate p-nitrophenyl ester hydrolysis by NPP1 and NPP3 by >90% and 23-43%, respectively, and only slightly affected (0-40%) hydrolysis of ATP by NTPDase1,2,3,8. Analogue 3 is the most potent NPP1 inhibitor currently known, Ki = 20 nM and IC50 = 0.39 µM. Analogue 2a is a selective NPP1 inhibitor with Ki = 685 nM and IC50 = 0.57 µM. Analogues 1-3 were found mostly to be nonagonists of P2Y1/P2Y2/P2Y11 receptors. Docking analogues 1-3 into the NPP1 model suggested that activity correlates with the number of H-bonds with binding site residues. In conclusion, we propose analogues 2a and 3 as highly promising NPP1 inhibitors.
Assuntos
Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/síntese química , Organofosfatos/síntese química , Organofosfonatos/síntese química , Organotiofosfatos/síntese química , Pirofosfatases/antagonistas & inibidores , Trifosfato de Adenosina/farmacologia , Animais , Sítios de Ligação , Células COS , Domínio Catalítico , Linhagem Celular Tumoral , Chlorocebus aethiops , Humanos , Ligação de Hidrogênio , Hidrólise , Simulação de Acoplamento Molecular , Organofosfatos/farmacologia , Organofosfonatos/farmacologia , Organotiofosfatos/farmacologia , Diester Fosfórico Hidrolases , Agonistas do Receptor Purinérgico P2Y/síntese química , Agonistas do Receptor Purinérgico P2Y/farmacologia , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
In this study we report the synthesis of C5/C6-fused uridine phosphonates that are structurally related to earlier reported allosteric P2Y2 receptor ligands. A silyl-Hilbert-Johnson reaction of six quinazoline-2,4-(1H,3H)-dione-like base moieties with a suitable ribofuranosephosphonate afforded the desired analogues after full deprotection. In contrast to the parent 5-(4-fluoropheny)uridine phosphonate, the present extended-base uridine phosphonates essentially failed to modulate the P2Y2 receptor.
Assuntos
Organofosfonatos/síntese química , Agonistas do Receptor Purinérgico P2Y/síntese química , Receptores Purinérgicos P2Y2/metabolismo , Uridina/síntese química , Regulação Alostérica , Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Linhagem Celular Tumoral , Humanos , Ligantes , Organofosfonatos/farmacologia , Agonistas do Receptor Purinérgico P2Y/farmacologia , Quinazolinas/química , Uridina/análogos & derivados , Uridina/farmacologiaRESUMO
A number of new amine scaffolds with good inhibitory activity in the ADP-induced platelet aggregation assay have been found to be potent antagonists of the P2Y1 receptor. SAR optimization led to the identification of isoindoline 3c and piperidine 4a which showed good in vitro binding and functional activities, as well as improved aqueous solubility. Among them, the piperidine 4a showed the best overall profile with favorable PK parameters.
Assuntos
Aminas/química , Agonistas do Receptor Purinérgico P2Y/química , Receptores Purinérgicos P2Y1/química , Ureia/análogos & derivados , Difosfato de Adenosina/farmacologia , Aminas/síntese química , Aminas/farmacocinética , Animais , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Meia-Vida , Humanos , Microssomos Hepáticos/metabolismo , Piperidinas/química , Inibidores da Agregação Plaquetária/síntese química , Inibidores da Agregação Plaquetária/química , Inibidores da Agregação Plaquetária/farmacocinética , Ligação Proteica , Agonistas do Receptor Purinérgico P2Y/síntese química , Agonistas do Receptor Purinérgico P2Y/farmacocinética , Ratos , Receptores Purinérgicos P2Y1/metabolismo , Relação Estrutura-Atividade , Ureia/síntese química , Ureia/farmacocinéticaRESUMO
We explored the influence of modifications of uridine 5'-methylenephosphonate on biological activity at the human P2Y(2) receptor. Key steps in the synthesis of a series of 5-substituted uridine 5'-methylenephosphonates were the reaction of a suitably protected uridine 5'-aldehyde with [(diethoxyphosphinyl)methylidene]triphenylphosphorane, C-5 bromination and a Suzuki-Miyaura coupling. These analogues behaved as selective agonists at the P2Y(2) receptor, with three analogues exhibiting potencies in the submicromolar range. Although maximal activities observed with the phosphonate analogues were much less than observed with UTP, high concentrations of the phosphonates had no effect on the stimulatory effect of UTP. These results suggest that these phosphonates bind to an allosteric site of the P2Y(2) receptor.
Assuntos
Organofosfonatos/química , Agonistas do Receptor Purinérgico P2Y/síntese química , Receptores Purinérgicos P2Y2/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Humanos , Organofosfonatos/síntese química , Organofosfonatos/farmacologia , Agonistas do Receptor Purinérgico P2Y/química , Agonistas do Receptor Purinérgico P2Y/farmacologia , Receptores Purinérgicos P2Y2/metabolismo , Nucleotídeos de Uracila/química , Uridina Trifosfato/metabolismoRESUMO
Dinucleoside polyphosphates, Np(n)N', exert their physiological effects via P2 receptors (P2Rs). Np(n)N' are attractive drug candidates as they offer better stability and specificity compared to nucleotides, the most common P2R ligands. To further improve the agonist properties of Np(n)N', we synthesized novel isosters of dinucleoside polyphosphates where N and N' are A or U and where the Pα or Pß phosphate groups are replaced by boranophosphate, denoted as Np(n)(α-B)N' or Np(n)(ß-B)N' (n = 3, 4), respectively. The potency of Np(n)(α/ß-B)N' analogues was evaluated at tP2Y(1), hP2Y(2), hP2Y(4), and rP2Y(6) receptors. The most potent P2Y(1)R and P2Y(6)R agonists were the Up(4)(ß-B)A (A isomer, EC(50) of 0.5 µM vs 0.004 µM for 2-SMe-ADP) and Up(3)(α-B)U (B isomer, EC(50) of 0.3 µM vs 0.2 µM for UDP), respectively. The receptor subtype selectivity is controlled by the position of the borano moiety on the Np(n)N' polyphosphate chain and the type of the nucleobase. In addition, Np(n)(α/ß-B)N' proved â¼22-fold more resistant to hydrolysis by e-NPP1, as compared to the corresponding Np(n)N' analogues. In summary, Up(4)(ß-B)A and Up(3)(α-B)U are potent, stable, and highly selective P2Y(1) and P2Y(6) receptor agonists, respectively.
Assuntos
Boranos/síntese química , Fosfatos de Dinucleosídeos/síntese química , Agonistas do Receptor Purinérgico P2Y/síntese química , Receptores Purinérgicos P2Y/metabolismo , Animais , Boranos/química , Boranos/farmacologia , Cálcio/metabolismo , Linhagem Celular Tumoral , Fosfatos de Dinucleosídeos/química , Fosfatos de Dinucleosídeos/farmacologia , Estabilidade de Medicamentos , Humanos , Hidrólise , Modelos Moleculares , Conformação Molecular , Diester Fosfórico Hidrolases/química , Agonistas do Receptor Purinérgico P2Y/química , Agonistas do Receptor Purinérgico P2Y/farmacologia , Pirofosfatases/química , Estereoisomerismo , Relação Estrutura-Atividade , TurquiaRESUMO
Nucleotide pyrophosphatase/phosphodiesterases (NPPs) hydrolyze extracellular nucleotides and dinucleotides and thus control purinergic signaling. Enhanced NPP activity is implicated in health disorders such as osteoarthritis and cancer. We designed novel diadenosine polyphosphonate derivatives as potential NPP inhibitors. Analogues 1-4 bear a phosphonate and/or boranophosphate group and/or a 2'-H atom instead of a 2'-OH group. In comparison to ATP, analogues 1-4 were barely hydrolyzed by human NTPDase1, -2, -3, and -8 (<5% hydrolysis) and NPP1 and -3 (≤ 13%) and were not hydrolyzed by ecto-5'-nucleotidase, unlike AMP. These derivatives did not affect NTPDase activity, and analogues 1 and 2 did not inhibit ecto-5'-nucleotidase. All analogues blocked â¼80% of the NPP2-dependent hydrolysis of pnp-TMP, a specific NPP substrate, and inhibited the catabolism of pnp-TMP (K(i) and IC50 both found to be between 10 and 60 µM), Ap5A, and ATP by NPP1. The activity of NPP3 was inhibited to a lesser extent by the new analogues, with compounds 1 and 4 being the most effective in that respect. The analogues dramatically reduced the level of hydrolysis of pnp-TMP at the cell surface of both osteocarcinoma and colon cancer cells. Importantly, analogues 1-4 exhibited significantly reduced agonistic activity toward human P2Y1,11) receptors (except for analogue 1) and no activity with human P2Y2 receptor. Our data provide strong evidence that analogue 2 is the first specific NPP inhibitor to be described.