RESUMEN
BACKGROUND: Indigo naturalis is a Traditional Chinese Medicine (TCM) ingredient long-recognized as a therapy for several inflammatory conditions, including psoriasis. However, its mechanism is unknown due to lack of knowledge about the responsible chemical entity. We took a different approach to this challenge by investigating the molecular profile of Indigo naturalis treatment and impacted pathways. METHODS: A randomized, double-blind, placebo-controlled clinical study was conducted using Indigo naturalis as topical monotherapy to treat moderate plaque psoriasis in a Chinese cohort (n = 24). Patients were treated with Indigo naturalis ointment (n = 16) or matched placebo (n = 8) twice daily for 8 weeks, with 1 week of follow-up. RESULTS: At week 8, significant improvements in Psoriasis Area and Severity Index (PASI) scores from baseline were observed in Indigo naturalis-treated patients (56.3% had 75% improvement [PASI 75] response) compared with placebo (0.0%). A gene expression signature of moderate psoriasis was established from baseline skin biopsies, which included the up-regulation of the interleukin (IL)-17 pathway as a key component; Indigo naturalis treatment resulted in most of these signature genes returning toward normal, including down-regulation of the IL-17 pathway. Using an in vitro keratinocyte assay, an IL-17-inhibitory effect was observed for tryptanthrin, a component of Indigo naturalis. CONCLUSIONS: This study demonstrated the clinical efficacy of Indigo naturalis in moderate psoriasis, and exemplified a novel experimental medicine approach to understand TCM targeting mechanisms. TRIAL REGISTRATION: NCT01901705 .
Asunto(s)
Indigofera/química , Interleucina-17/inmunología , Extractos Vegetales/administración & dosificación , Psoriasis/tratamiento farmacológico , Adulto , Anciano , Femenino , Humanos , Interleucina-17/genética , Masculino , Persona de Mediana Edad , Psoriasis/genética , Psoriasis/inmunología , Resultado del Tratamiento , Adulto JovenRESUMEN
BACKGROUND: Little is known about the impact of long-term use of immunosuppressive agents on immune response. OBJECTIVES: Assess the impact of continuous maintenance ustekinumab treatment on patients' ability to mount immune responses to pneumococcal (T-cell-independent) and tetanus toxoid (T-cell-dependent) vaccines. PATIENTS AND METHODS: Ustekinumab-treated patients with moderate-to-severe psoriasis treated in the long-term extension of the Phase 3 PHOENIX 2 trial (n=60) were compared with control psoriasis patients not receiving systemic therapy (n=56). Patients were vaccinated with both 23-valent pneumococcal and tetanus toxoid vaccines. Serum samples collected pre-vaccination and 4 weeks post-vaccination were assessed for antibody responses. RESULTS: No differences in the ability of ustekinumab-treated patients to respond to pneumococcal or tetanus toxoid vaccinations were observed compared with controls. A ≥2-fold increase in antibody levels in ≥7 of 14 serotypes of the pneumococcal vaccine was observed in ustekinumab-treated (96.6%) and untreated control (92.6%) patients following vaccination. Ustekinumab-treated patients achieved a ≥4-fold increase (84.7%) in anti-tetanus antibody vs. 77.8% in the control group. No differences were detected in ex-vivo responses to anti-CD3/CD28 or tetanus toxoid between ustekinumab-treated and control groups. CONCLUSION: Long-term treatment (≥3 years) with ustekinumab does not compromise the immune response to T-cell-dependent/-independent vaccines in patients with moderate-to-severe psoriasis.
Asunto(s)
Anticuerpos Monoclonales Humanizados/efectos adversos , Psoriasis/inmunología , Streptococcus pneumoniae/inmunología , Toxoide Tetánico/inmunología , Adulto , Anticuerpos/análisis , Anticuerpos Monoclonales Humanizados/uso terapéutico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Vacunas Neumococicas/inmunología , Psoriasis/tratamiento farmacológico , Ustekinumab , VacunaciónRESUMEN
Conformational analysis of the 3-benzylpiperidine in CCR3 antagonist clinical candidate 1 (BMS-639623) predicts that the benzylpiperidine may be replaced by acyclic, conformationally stabilized, anti-1,2-disubstituted phenethyl- and phenpropylamines. Ab initio calculations, enantioselective syntheses, and evaluation in CCR3 binding and chemotaxis assays of anti-1-methyl-2-hydroxyphenethyl- and phenpropylamine-containing CCR3 antagonists support this conformational correlation.
Asunto(s)
Piperidinas/química , Piperidinas/farmacología , Receptores CCR3/antagonistas & inhibidores , Urea/análogos & derivados , Ciclización , Enlace de Hidrógeno , Conformación Molecular , Urea/química , Urea/farmacologíaRESUMEN
Conformational analysis of trans-1,2-disubstituted cyclohexane CCR3 antagonist 2 revealed that the cyclohexane linker could be replaced by an acyclic syn-alpha-methyl-beta-hydroxypropyl linker. Synthesis and biological evaluation of mono- and disubstituted propyl linkers support this conformational correlation. It was also found that the alpha-methyl group to the urea lowered protein binding and that the beta-hydroxyl group lowered affinity for CYP2D6. Ab initio calculations show that the alpha-methyl group governs the spatial orientation of three key functionalities within the molecule. alpha-Methyl-beta-hydroxypropyl urea 31 with a chemotaxis IC(50)=38 pM for eosinophils was chosen to enter clinical development for the treatment of asthma.
Asunto(s)
Quimiotaxis de Leucocito/efectos de los fármacos , Eosinófilos/efectos de los fármacos , Piperidinas/farmacología , Receptores CCR3/antagonistas & inhibidores , Urea/análogos & derivados , Administración Oral , Animales , Inhibidores Enzimáticos del Citocromo P-450 , Perros , Eosinófilos/citología , Enlace de Hidrógeno , Ratones , Conformación Molecular , Piperidinas/química , Piperidinas/farmacocinética , Ratas , Relación Estructura-Actividad , Urea/química , Urea/farmacocinética , Urea/farmacologíaRESUMEN
Starting with our previously described(20) class of CC chemokine receptor-3 (CCR3) antagonist, we improved the potency by replacing the phenyl linker of 1 with a cyclohexyl linker and by replacing the 4-benzylpiperidine with a 3-benzylpiperidine. The resulting compound, 32, is a potent and selective antagonist of CCR3. SAR studies showed that the 3-acetylphenyl urea of 32 could be replaced with heterocyclic ureas or heterocyclic-substituted phenyl ureas and still maintain the potency (inhibition of eotaxin-induced chemotaxis) of this class of compounds in the low-picomolar range (IC(50) = 10-60 pM), representing some of the most potent CCR3 antagonists reported to date. The potency of 32 for mouse CCR3 (chemotaxis IC(50) = 41 nM) and its oral bioavailability in mice (20% F ) were adequate to assess the efficacy in animal models of allergic airway inflammation. Oral administration of 32 reduced eosinophil recruitment into the lungs in a dose-dependent manner in these animal models. On the basis of its overall potency, selectivity, efficacy, and safety profile, the benzenesulfonate salt of 32, designated DPC168, entered phase I clinical trials.
Asunto(s)
Ciclohexanos/síntesis química , Compuestos de Fenilurea/síntesis química , Piperidinas/síntesis química , Receptores de Quimiocina/antagonistas & inhibidores , Animales , Disponibilidad Biológica , Células CHO , Células CACO-2 , Calcio/metabolismo , Quimiotaxis de Leucocito/efectos de los fármacos , Cricetinae , Ciclohexanos/química , Ciclohexanos/farmacología , Eosinófilos/efectos de los fármacos , Eosinófilos/fisiología , Femenino , Humanos , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Técnicas In Vitro , Inflamación/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Permeabilidad , Compuestos de Fenilurea/química , Compuestos de Fenilurea/farmacología , Piperidinas/química , Piperidinas/farmacología , Receptores CCR3 , Estereoisomerismo , Relación Estructura-Actividad , Urea/análogos & derivados , Urea/síntesis química , Urea/química , Urea/farmacologíaRESUMEN
DPC168, a benzylpiperidine-substituted aryl urea CCR3 antagonist evaluated in clinical trials, was a relatively potent inhibitor of the 2D6 isoform of cytochrome P-450 (CYP2D6). Replacement of the cyclohexyl central ring with saturated heterocycles provided potent CCR3 antagonists with improved selectivity against CYP2D6. The favorable preclinical profile of DPC168 was maintained in an acetylpiperidine derivative, BMS-570520.
Asunto(s)
Compuestos de Bencilo/química , Compuestos de Bencilo/farmacología , Inhibidores del Citocromo P-450 CYP2D6 , Compuestos de Fenilurea/química , Piperidinas/química , Piperidinas/farmacología , Receptores de Quimiocina/antagonistas & inhibidores , Animales , Compuestos de Bencilo/síntesis química , Bioensayo , Células Cultivadas , Humanos , Ratones , Pan troglodytes , Compuestos de Fenilurea/farmacología , Piperidinas/síntesis química , Receptores CCR3 , Relación Estructura-ActividadRESUMEN
Linear unselective CCR3 antagonist leads with IC(50) values in the 200 nM range were converted into low nM binding compounds selective at CCR3 by moving the piperidine nitrogen substituent to the carbon at the 2-position of the ring. Substitution of the piperidine nitrogen with simple alkyl and acyl groups was found to improve the selectivity of this new compound class. In particular, N-{3-[(2S, 4R)-1-(propyl)-4-(4-fluorobenzyl)piperidinyl]propyl}-N'-(3-acetylphenyl)urea exhibited single digit nanomolar IC(50) values for CCR3 with >100-fold selectivity against an extensive counter screen panel.
Asunto(s)
Piperidinas/síntesis química , Piperidinas/farmacología , Receptores de Quimiocina/antagonistas & inhibidores , Piperidinas/química , Receptores CCR3 , Relación Estructura-ActividadRESUMEN
CC chemokine receptor (CCR) 3 is a chemokine receptor implicated in recruiting cells, particularly eosinophils (EPhi), to the lung in episodes of allergic asthma. To investigate the efficacy of selective, small molecule antagonists of CCR3, we developed a murine model of EPhi recruitment to the lung. Murine eotaxin was delivered intranasally to mice that had previously received i.p. injections of ovalbumin (OVA), and the effects were monitored by bronchoalveolar lavage. A selective eosinophilic influx was produced in animals receiving eotaxin but not saline. Furthermore, the number of EPhi was concentration- and time-dependent. Although anti-CCR3 antibody reduced the number of EPhi, the effect of eotaxin in OVA-sensitized mice was not a direct chemotactic stimulus because mast cell deficiency (in WBB6F1-Kitw/Kitw-v mice) significantly reduced the response. Two representative small molecule CCR3 antagonists from our program were characterized as being active at mouse CCR3. They were administered p.o. to wild-type mice and found to reduce eotaxin-elicited EPhi selectively in a dose-dependent manner. Pump infusion of one of the inhibitors to achieve steady-state levels showed that efficacy was not achieved at plasma concentrations equivalent to the in vitro chemotaxis IC90 but only at much higher concentrations. To extend the results from our recruitment model, we tested one of the inhibitors in an allergenic model of airway inflammation, generated by adoptive transfer of OVA-sensitive murine T helper 2 cells and aerosolized OVA challenge of recipient mice, and found that it inhibited EPhi recruitment. We conclude that small molecule CCR3 antagonists reduce pulmonary eosinophilic inflammation elicited by chemokine or allergenic challenge.
Asunto(s)
Inhibición de Migración Celular , Modelos Animales de Enfermedad , Eosinófilos/metabolismo , Pulmón/metabolismo , Receptores de Quimiocina/antagonistas & inhibidores , Hipersensibilidad Respiratoria/metabolismo , Animales , Células CHO , Cricetinae , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Femenino , Humanos , Inflamación/inmunología , Inflamación/metabolismo , Pulmón/inmunología , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Receptores CCR3 , Receptores de Quimiocina/inmunología , Receptores de Quimiocina/metabolismo , Hipersensibilidad Respiratoria/inmunologíaRESUMEN
The synthesis and structure-activity relationships of N-arylalkylpiperidylmethyl ureas as antagonists of the CC chemokine receptor-3 (CCR3) are presented. These compounds displayed potent binding to the receptor as well as functional antagonism of eotaxin-elicited effects on eosinophils.
Asunto(s)
Piperidinas/síntesis química , Receptores de Quimiocina/antagonistas & inhibidores , Urea/síntesis química , Señalización del Calcio/efectos de los fármacos , Quimiocina CCL11 , Quimiocinas CC/farmacología , Interacciones Farmacológicas , Eosinófilos/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Piperidinas/farmacología , Unión Proteica , Receptores CCR3 , Relación Estructura-Actividad , Urea/farmacologíaRESUMEN
The discovery of novel and selective small molecule antagonists of the CC Chemokine Receptor-3 (CCR3) is presented. Simple conversion from a 4- to 3-benzylpiperidine gave improved selectivity for CCR3 over the serotonin 5HT(2A) receptor. Chiral resolution and exploration of mono- and disubstitution of the N-propylurea resulted in several 3-benzylpiperidine N-propylureas with CCR3 binding IC(50)s under 5 nM. Data from in vitro calcium mobilization and chemotaxis assays for these compounds ranged from high picomolar to low nanomolar EC(50)s and correlated well with antagonist binding IC(50)s.
Asunto(s)
Piperidinas/metabolismo , Receptores de Quimiocina/antagonistas & inhibidores , Receptores de Quimiocina/metabolismo , Urea/análogos & derivados , Urea/metabolismo , Animales , Células CHO , Bovinos , Cricetinae , Piperidinas/química , Unión Proteica/fisiología , Receptores CCR3 , Urea/químicaRESUMEN
CCR3 antagonist leads with IC(50) values in the microM range were converted into low nM binding compounds that displayed in vitro inhibition of human eosinophil chemotaxis induced by human eotaxin. In particular, 4-benzylpiperidin-1-yl-n-propylureas and erythro-3-(4-benzyl-2-(alpha-hydroxyalkyl)piperidin-1-yl)-n-propylureas (obtained via Beak reaction of N-BOC-4-benzylpiperidine) exhibited single digit nanomolar IC(50) values for CCR3.