Substituted indazole derivatives and their use as inhibitors of PDE4 and the production of tumour necrosis factor

This patent discloses indazole derivatives that exhibit activity as inhibitors of phosphodiesterase Type 4 (PDE4) and also inhibit the production of tumour necrosis factor (TNF). These compounds are expected to have utility in the treatment of inflammatory diseases such as asthma and rheumatoid arthritis.     

An efficient large‐scale synthesis of 1H‐indazole‐[3‐14C]carboxylic acid

An efficient large‐scale carbon‐14 synthesis of 1H‐indazole‐[3‐¹⁴C]carboxylic acid starting from [¹⁴C]potassium cyanide is reported. Key transformations encountered during the synthesis include aromatic nucleophilic substitution of 2‐nitrofluorobenzene by ethyl [¹⁴C]cyanoacetate, a mild decarbethoxylation and an aniline nitrosation/cyclization. Copyright © 2007 John Wiley & Sons, Ltd.     

Substituted alkylamines and methods of use

This Amgen patent application provides a series of 2-aminoheterocyclic amides described as useful for the treatment of cancer and angiogenesis-related disorders. These compounds appear to be related to a nicotinamide series previously described by Novartis as vascular endothelial growth factor (VEGF) inhibitors. The compounds differ from the Novartis series in that they rely preferentially on an aminobicycloaromatic group at the 2-position of the nicotinamide. Based on the data provided in the application, a 6-amino-(1H) indazole appears to be a particularly important sidechain for these compounds, providing consistent cell activity despite a range of substituted amides at the 3-position of the nicotinamide.     

Identification of Potent Reverse Indazole Inhibitors for HPK1

Hematopoietic progenitor kinase (HPK1), a negative regulator of TCR-mediated T-cell activation, has been recognized as a novel antitumor immunotherapy target. Structural optimization of kinase inhibitor 4 through a systematic two-dimensional diversity screen of pyrazolopyridines led to the identification of potent and selective compounds. Crystallographic studies with HPK1 revealed a favorable water-mediated interaction with Asp155 and a salt bridge to Asp101 with optimized heterocyclic solvent fronts that were critical for enhanced potency and selectivity. Computational studies of model systems revealed differences in torsional profiles that allowed for these beneficial protein–ligand interactions. Further optimization of molecular properties led to identification of potent and selective reverse indazole inhibitor 36 that inhibited phosphorylation of adaptor protein SLP76 in human PBMC and exhibited low clearance with notable bioavailability in in vivo rat studies.     

Identification of three cannabimimetic indazole and pyrazole derivatives,APINACA 2H‐indazole analogue,AMPPPCA, and 5F‐AMPPPCA

This paper reports analytical properties of three cannabimimetic indazole and pyrazole derivatives seized from a clandestine laboratory. These three new synthetic cannabinoids include N ‐(1‐adamantyl)‐2‐pentyl‐2H ‐indazole‐3‐carboxamide (APINACA 2H ‐indazole analogue, 1 ), N ‐(1‐adamantyl)‐4‐methyl‐1‐pentyl‐5‐phenyl‐1H ‐pyrazole‐3‐carboxamide (AMPPPCA, 2 ), and N ‐(1‐adamantyl)‐1‐(5‐fluoropentyl)‐4‐methyl‐5‐phenyl‐1H ‐pyrazole‐3‐carboxamide (5F‐AMPPPCA, 3 ). These compounds were identified by liquid chromatography‐quadrupole time‐of‐flight‐mass spectrometry (LC‐QTOF‐MS), gas chromatography‐time‐of‐flight‐mass spectrometry (GC‐TOF‐MS), and nuclear magnetic resonance (NMR) spectroscopy. No analytical properties and pharmacological activities about compounds 1–3 have appeared until now, making this the first report on these compounds. Copyright © 2016 John Wiley & Sons, Ltd.     

吲哚并吲哒唑类衍生物对人脑胶质瘤U251细胞生长的抑制作用

 目的 研究6-甲基-11-(4-二甲基氨基苄烯-7,8,9,10-四氢-1-氢吲哚［1,2b］吲哒唑三氟甲磺酸盐对人脑胶质瘤U251细胞增殖,凋亡和细胞周期的影响。方法 分别采用噻唑蓝(MTT法检测U251细胞增殖活性、Hoechst33258荧光染色检测细胞凋亡形态、流式细胞仪检测细胞周期、Western blot检测细胞周期蛋白Cyclin D1和Cyclin B1的表达变化。结果 MTT法检测发现6-甲基-11-(4-二甲基氨基苄烯-7,8,9,10-四氢-1-氢吲哚［1,2b］吲哒唑三氟甲磺酸盐能显著抑制U251细胞的生长,且呈浓度及时间依赖性, Hoechst33258荧光染色检测观察到细胞凋亡形态的改变,流式细胞仪检测发现细胞阻滞于G2/M期和S 期,Western blot检测发现细胞周期蛋白Cyclin D1和Cyclin B1表达下调。结论 6-甲基-11-(4-二甲基氨基苄烯-7,8,9,10-四氢-1-氢吲哚［1,2b］吲哒唑三氟甲磺酸盐对人脑胶质瘤U251细胞增殖有明显的抑制作用,并能诱导其发生凋亡。其凋亡作用可能与G2/M期和S期阻滞有关。     

Nitric oxide synthase inhibition attenuates saccharin or ethanol reinforced responding in long-evans rats

1. 1. Effect of 7-nitro indazole (7-NI), brain specific inhibitor of nitric oxide synthase (NOS), on saccharin or ethanol reinforcement was investigated in male Long-Evans rats.

2. 2. Twenty-four rats were trained to lever press both for a 1-% (w/v) saccharin solution and 10% (v/v) ethanol solution using a lever-pressing schedule of progressive ratio.

3. 3. Well-trained rats were tested with 7-NI (10, 20 and 40 mg/kg) or a vehicle (methylcellulose, 2%) injected ip. 60 min before each test. In some tests, L-arginine (1000 mg/kg, ip), a NO precursor, was injected 30 min before a 7-NI (40 mg/kg) injection.

4. 4. Treatment with 7-NI reduced the lever pressing for either saccharin or ethanol reinforcement in a dose dependent manner. The inhibitory effects of 7-NI on the lever pressing for saccharin or ethanol were blocked by L-arginine pre-administration.

5. 5. Our results suggest that reduction in ethanol reinforcement by NOS inhibition may be related to central mechanisms modulating feeding behavior in rats.

     

Effect of YC-1, an NO-independent,superoxide-sensitive stimulator of soluble guanylyl cyclase,on smooth muscle responsiveness to nitrovasodilators

1. We studied the effects of 3-(5′-hydroxymethyl-2′furyl)-1-benzyl indazole (YC-1) on the activity of purified soluble guanylyl cyclase (sGC), the formation of guanosine-3′ : 5′ cyclic monophosphate (cyclic GMP) in vascular smooth muscle cells (VSMC), and on the tone of rabbit isolated aortic rings preconstricted by phenylephrine (PE). In addition, we assessed the combined effect of YC-1, and either NO donors, or superoxide anions on these parameters.
2. YC-1 elicited a direct concentration-dependent activation of sGC (EC₅₀ 18.6±2.0 μM), which was rapid in onset and quickly reversible upon dilution. YC-1 altered the enzyme kinetics with respect to GTP by decreasing K_M and increasing V_max. Activation of sGC by a combination of sodium nitroprusside (SNP) and YC-1 was superadditive at low and less than additive at high concentrations, indicating a synergistic activation of the enzyme by both agents. A specific inhibitor of sGC, 1H-(1,2,4)-oxdiazolo-(4,3-a)-6-bromo-quinoxazin-1-one (NS 2028), abolished activation of the enzyme by either compound.
3. YC-1 induced a concentration-dependent increase in intracellular cyclic GMP levels in rat cultured aortic VSMC, which was completely inhibited by NS 2028. YC-1 applied at the same concentration as SNP elicited 2.5 fold higher cyclic GMP formation. Cyclic GMP-increases in response to SNP and YC-1 were additive.
4. YC-1 relaxed preconstricted endothelium-denuded rabbit aortic rings in a concentration-dependent manner (50% at 20 μM) and markedly increased cyclic GMP levels. Relaxations were inhibited by NS 2028. A concentration of YC-1 (3 μM), which elicited only minor effects on relaxation and cyclic GMP, increased the vasodilator potency of SNP and nitroglycerin (NTG) by 10 fold and markedly enhanced SNP- and NTG-induced cyclic GMP formation.
5. Basal and YC-1-stimulated sGC activity was sensitive to inhibition by superoxide (O₂⁻) generated by xanthine/xanthine oxidase, and was protected from this inhibition by superoxide dismutase (SOD). YC-1-stimulated sGC was also sensitive to inhibition by endogenously generated (O₂⁻ in rat preconstricted endothelium-denuded aortic rings. Relaxation to YC-1 was significantly attenuated in aortae from spontaneously hypertensive rats (SHR), which generated O₂⁻ at a higher rate than aortae from normotensive Wistar Kyoto rats (WKY). SOD restored the vasodilator responsiveness of SHR rings to YC-1.
6. In conclusion, these results indicate that YC-1 is an NO-independent, O₂⁻-sensitive, direct activator of sGC in VSMC and exerts vasorelaxation by increasing intracellular cyclic GMP levels. The additive or even synergistic responses to NO-donors and YC-1 in cultured VSMC and isolated aortic rings apparently reflect the direct synergistic action of YC-1 and NO on the sGC. The synergism revealed in this in vitro study suggests that low doses of YC-1 may be of therapeutic value by permitting the reduction of nitrovasodilator dosage.

     

Alleviation of extensive visual pathway dysfunction by a remyelinating drug in a chronic mouse model of multiple sclerosis

Visual deficits are among the most prevalent symptoms in patients with multiple sclerosis (MS). To understand deficits in the visual pathway during MS and potential treatment effects, we used experimental autoimmune encephalomyelitis (EAE), the most commonly used animal model of MS. The afferent visual pathway was assessed in vivo using optical coherence tomography (OCT), electroretinography (ERG), and visually evoked cortical potentials (VEPs). Inflammation, demyelination, and neurodegeneration were examined by immunohistochemistry ex vivo. In addition, an immunomodulatory, remyelinating agent, the estrogen receptor β ligand chloroindazole (IndCl), was tested for its therapeutic potential in the visual pathway. EAE produced functional deficits in visual system electrophysiology, including suppression of ERG and VEP waveform amplitudes and increased signal latencies. Therapeutic IndCl rescued overall visual system latency by VEP but had little impact on amplitude or ERG findings relative to vehicle. Faster VEP conduction in IndCl‐treated mice was associated with enhanced myelin basic protein signal in all visual system structures examined. IndCl preserved retinal ganglion cells (RGCs) and oligodendrocyte density in the prechiasmatic white matter, but similar retinal nerve fiber layer thinning by OCT was noted in vehicle and IndCl‐treated mice. Although IndCl differentially attenuated leukocyte and astrocyte staining signal throughout the structures analyzed, axolemmal varicosities were observed in all visual fiber tracts of mice with EAE irrespective of treatment, suggesting impaired axonal energy homeostasis. These data support incomplete functional recovery of VEP amplitude with IndCl, as fiber tracts displayed persistent axon pathology despite remyelination‐induced decreases in latencies, evidenced by reduced optic nerve g‐ratio in IndCl‐treated mice. Although additional studies are required, these findings demonstrate the dynamics of visual pathway dysfunction and disability during EAE, along with the importance of early treatment to mitigate EAE‐induced axon damage.     

(WO2011057959) Indole and indazole derivatives as glycogen synthase activators: a patent evaluation

Scientific evaluation of a patent aiming for the development of indole and indazole derivatives from biaryloxymethylarene as glycogen synthase activators, a key enzyme involved in type 2 diabetes mellitus.