English:
[0003] Voltage gated Na channels comprise a gene family consisting of 9 different subtypes (NaV 1.1 -NaVl .9). As shown in Table A below these subtypes show tissue specific localization and functional differences (See, Goldin, A. L. (2001) "Resurgence of sodium channel research" Annu Rev Physiol 63: 871-94). Three members of the gene family (NaVl .8, 1.9, 1.5) are resistant to block by the well-known Na channel blocker TTX, demonstrating subtype specificity within this gene family. Mutational analysis has identified glutamate 387 as a critical residue for TTX binding (See, Noda, M., H. Suzuki, et al. (1989) "A single point mutation confers tetrodotoxin and saxitoxin insensitivity on the sodium channel II" FEBS Lett 259(1): 213-6).
[0004] Table A (Abbreviations: CNS = central nervous system, PNS = peripheral nervous sytem, DRG = dorsal root ganglion, TG = Trigeminal ganglion):
[0005] In general, voltage-gated sodium channels (NaVs) are responsible for initiating the rapid upstroke of action potentials in excitable tissue in nervous system, which transmit the electrical signals that compose and encode normal and aberrant pain sensations. Antagonists of NaV channels can attenuate these pain signals and are useful for treating a variety of pain conditions, including but not limited to acute, chronic, inflammatory, and neuropathic pain. Known NaV antagonists, such as TTX, lidocaine (See, Mao, J. and L. L. Chen (2000) "Systemic lidocaine for neuropathic pain relief Pain 87(1): 7-17.) bupivacaine, phenytoin (See. Jensen, T. S. (2002) "Anticonvulsants in neuropathic pain: rationale and clinical evidence" Eur J Pain 6 (Suppl A): 61-8), lamotrigine (See, Rozen, T. D. (2001) "Antiepileptic drugs in the management of cluster headache and trigeminal neuralgia" Headache 41 Suppl 1: S25-32 and Jensen, T. S. (2002) "Anticonvulsants in neuropathic pain: rationale and clinical evidence" Eur J Pain 6 (Suppl A): 61-8.), and carbamazepine (See, Backonja, M. M. (2002) "Use of anticonvulsants for treatment of neuropathic pain" Neurology 59(5 Suppl 2): S 14-7), have been shown to be useful attenuating pain in humans and animal models.
[0006] Hyperalgesia (extreme sensitivity to something painful) that develops in the presence of tissue injury or inflammation reflects, at least in part, an increase in the excitability of high-threshold primary afferent neurons innervating the site of injury. Voltage sensitive sodium channels activation is critical for the generation and propagation of neuronal action potentials. There is a growing body of evidence indicating that modulation of NaV currents is an endogenous mechanism used to control neuronal excitability (See, Goldin, A. L. (2001) "Resurgence of sodium channel research" Annu Rev Physiol 63: 871-94.). Several kinetically and pharmacologically distinct voltage-gated sodium channels are found in dorsal root ganglion (DRG) neurons. The TTX-resistant current is insensitive to micromolar concentrations of tetrodotoxin, and displays slow activation and inactivation kinetics and a more depolarized activation threshold when compared to other voltage-gated sodium channels. TTX-resistant sodium currents are primarily restricted to a subpopulation of sensory neurons likely to be involved in nociception. Specifically, TTX-resistant sodium currents are expressed almost exclusively in neurons that have a small cell-body diameter; and give rise to small-diameter slow-conducting axons and that are responsive to capsaicin. A large body of experimental evidence demonstrates that TTX-resistant sodium channels are expressed on C-fibers and are important in the transmission of nociceptive information to the spinal cord.
[0007] Intrathecal administration of antisense oligo-deoxynucleotides targeting a unique region of the TTX-resistant sodium channel (NaVl.8) resulted in a significant reduction in PGE2-induced hyperalgesia (See, Khasar, S. G., M. S. Gold, et al. (1998) "A tetrodotoxin-resistant sodium current mediates inflammatory pain in the rat" Neurosci Lett 256(1): 17-20). More recently, a knockout mouse line was generated by Wood and colleagues, which lacks functional NaVl.8. The mutation has an analgesic effect in tests assessing the animal's response to the inflammatory agent carrageenan (See, Akopian, A. N., V. Souslova, et al. (1999) "The tetrodotoxin-resistant sodium channel SNS has a specialized function in pain pathways" Nat Neurosci 2(6): 541-8.). In addition, deficit in both mechano- and thermoreception were observed in these animals. The analgesia shown by the Navl .8 knockout mutants is consistent with observations about the role of TTX-resistant currents in nociception.
[0008] Immunohistochemical, in-situ hybridization and in-vitro electrophysiology experiments have all shown that the sodium channel NaVl.8 is selectively localized to the small sensory neurons of the dorsal root ganglion and trigeminal ganglion (See, Akopian, A. N., L. Sivilotti, et al. (1996) "A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons" Nature 379(6562): 257-62.). The primary role of these neurons is the detection and transmission of nociceptive stimuli. Antisense and immunohistochemical evidence also supports a role for NaVl .8 in neuropathic pain (See, Lai, J., M. S. Gold, et al. (2002) "Inhibition of neuropathic pain by decreased expression of the tetrodotoxin-resistant sodium channel, NaVl .8" Pain 95(1-2): 143-52, and Lai, J., J. C. Hunter, et al. (2000) "Blockade of neuropathic pain by antisense targeting of tetrodotoxin- resistant sodium channels in sensory neurons" Methods Enzymol 314: 201-13.). NaVl.8 protein is upregulated along uninjured C-fibers adjacent to the nerve injury. Antisense treatment prevents the redistribution of NaVl .8 along the nerve and reverses neuropathic pain. Taken together the gene-knockout and antisense data support a role for NaVl .8 in the detection and transmission of inflammatory and neuropathic pain.
[0009] In neuropathic pain states there is a remodeling of Na channel distribution and subtype. In the injured nerve, expression of NaVl .8 and NaVl .9 are greatly reduced whereas expression of the TTX sensitive subunit NaV1.3 is 5-10 fold upregulated (See, Dib-Haiύ S. D., J. Fjell, et al. (1999) "Plasticity of sodium channel expression in DRG neurons in the chronic constriction injury model of neuropathic pain." Pain 83(3): 591-600.) The timecourse of the increase in NaVl.3 parallels the appearance of allodynia in animal models subsequent to nerve injury. The biophysics of the NaVl .3 channel is distinctive in that it shows very fast repriming after inactivation following an action potential. This allows for sustained rates of high firing as is often seen in the injured nerve (See, Cummins, T. R., F. Aglieco, et al. (2001) "Navl.3 sodium channels: rapid repriming and slow closed-state inactivation display quantitative differences after expression in a mammalian cell line and in spinal sensory neurons" J Neurosci 21(16): 5952-61.). NaVl.3 is expressed in the central and peripheral systems of man. NaVl .9 is similar to NaVl .8 as it is selectively localized to small sensory neurons of the dorsal root ganglion and trigeminal ganglion (See, Fang, X., L. Djouhri, et al. (2002). "The presence and role of the tetrodotoxin-resistant sodium channel Na(v)l .9 (NaN) in nociceptive primary afferent neurons." J Neurosci 22(17): 7425-33.). It has a slow rate of inactivation and left-shifted voltage dependence for activation (See, Dib- Hajj, S., J. A. Black, et al. (2002) "NaN/Navl.9: a sodium channel with unique properties" Trends Neurosci 25(5): 253-9.). These two biophysical properties allow NaVl .9 to play a role in establishing the resting membrane potential of nociceptive neurons. The resting membrane potential of NaVl.9 expressing cells is in the —55 to — 5OmV range compared to — 65mV for most other peripheral and central neurons. This persistent depolarization is in large part due to the sustained low-level activation of NaVl .9 channels. This depolarization allows the neurons to more easily reach the threshold for firing action potentials in response to nociceptive stimuli. Compounds that block the NaVl.9 channel may play an important role in establishing the set point for detection of painful stimuli. In chronic pain states, nerve and nerve ending can become swollen and hypersensitive exhibiting high frequency action potential firing with mild or even no stimulation. These pathologic nerve swellings are termed neuromas and the primary Na channels expressed in them are NaVl .8 and NaVl .7 (See, Kretschmer. T., L. T. Happel, et al. (2002) "Accumulation of PNl and PN3 sodium channels in painful human neuroma- evidence from immunocytochemistry" Acta Neurochir (Wien) 144(8): 803-10; discussion 810.). NaVl .6 and NaVl .7 are also expressed in dorsal root ganglion neurons and contribute to the small TTX sensitive component seen in these cells. NaVl .7 in particular my therefore be a potential pain target in addition to it's role in neuroendocrine excitability (See, Klugbauer, N., L. Lacinova, et al. (1995) "Structure and functional expression of a new member of the tetrodotoxin- sensitive voltage-activated sodium channel family from human neuroendocrine cells" Embo J 14(6): 1084-90). [0010] NaVl.1 (See, Sugawara, T., E. Mazaki-Miyazaki, et al. (2001) "Navl.l mutations cause febrile seizures associated with afebrile partial seizures." Neurology 57(4): 703-5.) and NaVl.2 (See. Sugawara, T., Y. Tsurubuchi, et al. (2001) "A missense mutation of the Na+ channel alpha II subunit gene Na(v)l .2 in a patient with febrile and afebrile seizures causes channel dysfunction" Proc Natl Acad Sci U S A 98(11): 6384-9) have been linked to epilepsy conditions including febrile seizures. There are over 9 genetic mutations in NaVl .1 associated with febrile seizures (See, Meisler, M. H., J. A. Kearney, et al. (2002) "Mutations of voltage-gated sodium channels in movement disorders and epilepsy" Novartis Found Svmp 241: 72-81)
[000368] In yet another embodiment of the method of the present invention, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 or 2 and at least one occurrence of R4is -SOR', -CON(R')2, -SO2N(R')2, -COR', or -COOR'. In another embodiment, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 and R4is -SOR'. In another embodiment, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 and R4is -COOR'. In another embodiment, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 and R4is -CON(R')2. In another embodiment, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 and R4is -SO2N(R')2- In another embodiment, R1and R2, taken together is optionally substituted piperazin-1-yl (dd), wherein z is 1 and R4is -COR'. [000369] In yet another embodiment of the method of the present invention, R1and R2, taken together is optionally substituted morpholin- 1 -yl (ee) or thiomorpholin-1 -yl (ff), wherein z is 1 or 2 and at least one occurrence of R4is -SOR', -CON(R')2, -SO2N(R')2, - COR', or -COOR'. In another embodiment, R1and R2, taken together is optionally substituted morpholin- 1-yl (ee) or thiomorpholin-1 -yl (ff), wherein z is 1 and R4is -SOR'. In another embodiment, R1and R2, taken together is optionally substituted morpholin- 1-yl (ee) or thiomorpholin-1-yl (ff), wherein z is 1 and R4is -COOR'. In another embodiment, R1and R2, taken together is optionally substituted morpholin- 1-yl (ee) or thiomorpholin-1-yl (ff), wherein z is 1 and R4is -CON(R')2- In another embodiment, R1and R2, taken together is optionally substituted morpholin- 1-yl (ee) or thiomorpholin-1-yl (ff), wherein z is 1 and R4is
[000409] 3-(2-Methoxybenzamido)thiophene-2-carboxamide.
[00090J In one embodiment, G3is N. Or, G3is CH.

中文:
[0003]电压-门控Na通道包含由9种不同的亚型组成的基因家族(NaV1.1-NaV1.9)。这些亚型显示组织特异性定位作用和功能差异(参见,Goldin,A.L.(2001)"Resurgenceofsodiumchannelresearch"AnnuRevPhysiol63:871-94)。该基因家族有三种成员(NaV1.8,1.9,1.5)耐受于被熟知的Na通道阻滞剂TTX所阻滞,证明在这种基因家族内存在亚型特异性。突变分析已经鉴别谷氨酸387是TTX结合的决定性残基(参见,Noda,M.,H.Suzuki,等人(1989)"AsinglepointmutationconferstetrodotoxinandsaxitoxininsensitivityonthesodiumchannelII"FEBSLett259(1):213-6)。
[0004]表A(缩写:CNS=中枢神经系统,PNS=外周神经系统,DRG=背根神经节,TG=三叉神经节):
[0005]一般而言,电压-门控钠通道(NaV)负责引发神经系统可兴奋组织作用电位的迅速提升,这传送编写和编码正常与异常疼痛感觉的电信号。NaV通道的拮抗剂能够减弱这些疼痛信号,可用于治疗多种疼痛病症,包括但不限于急性、慢性、炎性和神经病性疼痛。已知的NaV拮抗剂,例如TTX、利多卡因(参见,Mao,J.andL.L.Chen(2000)"Systemiclidocaineforneuropathicpainrelief"Pain87(1):7-17)、布比卡因、苯妥英(参见,Jensen,T.S.(2002)"Anticonvulsantsinneuropathicpain:rationaleandclinicalevidence"EurJPain6(SupplA):61-8)、拉莫三嗪(参见,Rozen,T.D.(2001)"Antiepilepticdrugsinthemanagementofclusterheadacheandtrigeminalneuralgia"Headache41Suppl1:S25-32andJensen,T.S.(2002)"Anticonvulsantsinneuropathicpain:rationaleandclinicalevidence"EurJPain6(SupplA):61-8)和卡马西平(参见,Backonja,M.M.(2002)"Useofanticonvulsantsfortreatmentofneuropathicpain"Neurology59(5Suppl2):S14-7),已经显示可用于减弱人和动物模型的疼痛。
[0006]在组织损伤或炎症的存在下形成的痛觉过敏(对一定的疼痛极度敏感)至少在部分程度上反映了支配损伤部位的高阈值主要传入神经元的兴奋性增加。电压敏感性钠通道活化对神经元作用电位的产生和传播而言是决定性的。越来越多的证据表明,NaV电流的调控是用于控制神经元兴奋性的内在机理(参见,Goldin,A.L.(2001)"Resurgenceofsodiumchannelresearch"AnnuRevPhysiol63:871-94)。在背根神经节(DRG)神经元中发现有若干动力学与药理学不同的电压-门控钠通道。TTX-耐受性电流对微摩尔浓度的河豚毒素不敏感,并且与其他电压-门控钠通道相比,表现缓慢的活化与失活动力学和更高的去极化活化阈。TTX-耐受性钠电流主要受到可能涉及伤害感受的感觉神经元亚群的限制。具体而言,TTX-耐受性钠电流几乎仅在细胞体直径小的神经元中被表达;引起小直径的、慢传导的轴突,并且对辣椒碱有响应。大量实验证据证明,TTX-耐受性钠通道在C-纤维上被表达,在感受脊髓伤害的信息传递中发挥重要作用。
[0007]靶向于TTX-耐受性钠通道独特区域(NaV1.8)的反义寡-脱氧核苷酸的鞘内给药导致PGE2-诱导的痛觉过敏的显著减少(参见,Khasar,S.G.,M.S.Gold,等人(1998)"Atetrodotoxin-resistantsodiumcurrentmediatesinflammatorypainintherat"NeurosciLett256(1):17-20)。最近,Wood和同事创建了一种剔除小鼠系,它缺乏功能性NaV1.8。在评估动物对致炎剂角叉菜胶的响应的试验中,该突变具有痛觉缺失效应(参见,Akopian,A.N.,V.Souslova,等人(1999)"Thetetrodotoxin-resistantsodiumchannelSNShasaspecializedfunctioninpainpathways"NatNeurosci2(6):541-8)。另外,在这些动物中观察到机械与温度感受的缺陷。由NaV1.8剔除突变体所显示的痛觉缺失与关于TTX-耐受性电流在伤害感受中的角色的观察结果是一致的。
[0008]免疫组织化学、就地杂交和体外电生理学实验都已显示,钠通道NaV1.8选择性地定位于背根神经节和三叉神经节的小感觉神经元(参见,Akopian,A.N.,L.Sivilotti,等人(1996)"Atetrodotoxin-resistantvoltage-gatedsodiumchannelexpressedbysensoryneurons"Nature379(6562):257-62)。这些神经元的主要角色是感受伤害刺激的检测和传递。反义与免疫组织化学证据也支持了NaV1.8在神经病性疼痛中的角色(参见,Lai,J.,M.S.Gold,等人(2002)"Inhibitionofneuropathicpainbydecreasedexpressionofthetetrodotoxin-resistantsodiumchannel,NaV1.8"Pain95(1-2):143-52,andLai,J.,J.C.Hunter,等人(2000)"Blockadeofneuropathicpainbyantisensetargetingoftetrodotoxin-resistantsodiumchannelsinsensoryneurons"MethodsEnzymol314:201-13)。NaV1.8蛋白沿着与神经损伤相邻的未损伤C-纤维被增量调节。反义处理预防NaV1.8沿着神经的再分布,逆转神经病性疼痛。综合基因-剔除和反义数据,支持了NaV1.8在炎性与神经病性疼痛的检测和传递中的角色。
[0009]在神经病性疼痛状态中,存在Na通道分布和亚型的改造。在受损的神经中,NaV1.8和NaV1.9的表达大大减少了,而TTX-敏感性亚单位NaV1.3的表达被5-10倍增量调节(参见,Dib-Hajj,S.D.,J.Fjell,等人(1999)"PlasticityofsodiumchannelexpressioninDRGneuronsinthechronicconstrictioninjurymodelofneuropathicpain."Pain83(3):591-600)。在继神经损伤之后的动物模型中,NaV1.3增加的时间过程平行于异常性疼痛的出现。NaV1.3的生物物理学的特有之处在于在动作电位后的失活之后显示非常快的重新引导。这产生持续的高发放速率,在受损的神经中经常见到(参见,Cummins,T.R.,F.Aglieco,等人(2001)"Nav1.3sodiumchannels:rapidreprimingandslowclosed-stateinactivationdisplayquantitativedifferencesafterexpressioninamammaliancelllineandinspinalsensoryneurons"JNeurosci21(16):5952-61)。NaV1.3在人的中枢与外周系统中被表达。NaV1.9与NaV1.8相似,它也选择性地定位于背根神经节和三叉神经节的小感觉神经元(参见,Fang,X.,L.Djouhri,等人(2002)"Thepresenceandroleofthetetrodotoxin-resistantsodiumchannelNa(v)1.9(NaN)innociceptiveprimaryafferentneurons."JNeurosci22(17):7425-33)。它具有缓慢的失活速率和就活化而言左移的电压依赖性(参见,Dib-Hajj,S.,J.A.Black,等人(2002)"NaN/Nav1.9:asodiumchannelwithuniqueproperties"TrendsNeurosci25(5):253-9)。这两种生物物理学性质允许NaV1.9在建立感受伤害神经元的静息膜电位中扮演角色。表达NaV1.9的细胞的静息膜电位在-55至-50mV范围内,大多数其他外周与中枢神经元为-65mV。这种持续性去极化在很大程度上是由于持续低水平的NaV1.9通道活化。这种去极化允许神经元更容易达到响应于感受伤害刺激而发放动作电位的阈值。阻滞NaV1.9通道的化合物可以在建立疼痛刺激检测的调定点中扮演重要角色。在慢性疼痛状态中,神经和神经末梢可能变得肿胀和过敏,即使对温和或甚至没有刺激也表现高频率的动作电位发放。这些病理性神经肿胀被称为神经瘤,在其中被表达的主要Na通道是NaV1.8和NaV1.7(参见,Kretschmer,T.,L.T.Happel,等人(2002)"AccumulationofPN1andPN3sodiumchannelsinpainfulhumanneuroma-evidencefromimmunocytochemistry"ActaNeurochir(Wien)144(8):803-10;discussion810)。NaV1.6和NaV1.7也在背根神经节神经元中被表达,并且对见于这些细胞中的小TTX-敏感性组分有贡献。除了它在神经内分泌兴奋性中的角色以外,NaV1.7特别因此还可能是潜在的疼痛靶(参见,Klugbauer,N.,L.Lacinova,等人(1995)"Structureandfunctionalexpressionofanewmemberofthetetrodotoxin-sensitivevoltage-activatedsodiumchannelfamilyfromhumanneuroendocrinecells"EmboJ14(6):1084-90)。
[000369]在本发明方法的另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1或2,至少一次出现的R4是-SOR′、-CON(R′)2、-SO2N(R′)2、-COR′或-COOR′。在另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1,R4是-SOR′。在另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1,R4是-COOR′。在另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1,R4是-CON(R′)2。在另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1,R4是-SO2N(R′)2。在另一种实施方式中,R1和R2一起是可选被取代的吗啉-1-基(ee)或硫吗啉-1-基(ff),其中z是1,R4是-COR′。
[000409]3-(2-甲氧基苯甲酰氨基)噻吩-2-酰胺。
[00090]在一种实施方式中,G3是N。或者,G3是CH。