公开(公告)号：US20120329723A1

公开(公告)日：2012-12-27

申请号：US13534576

申请日：2012-06-27

Applicant: Roger A. Johns ,  Feng Tao

Inventor： Roger A. Johns ,  Feng Tao

IPC: A61K38/17 ,  A61P25/20 ,  A61P25/04 ,  A61P23/00 ,  C07K19/00 ,  A61P25/00

CPC classification number: A61K38/1709 ,  C07K14/47 ,  C07K2319/10 ,  C07K2319/21 ,  C07K2319/23 ,  C07K2319/70 ,  C12N9/1229

Abstract: Cell-permeant fusion peptides Tat-PDZ can dose-dependently reduce the threshold for anesthesia. PDZ domain-mediated protein interactions at synapses in the central nervous system play an important role in the molecular mechanisms of anesthesia. Moreover, Tat-PDZ cell-permeant fusion peptides are delivered intracellularly into neurons in the central nervous system subsequent to intraperitoneally injection. By in vitro and in vivo binding assays, we found that the Tat-PDZ dose-dependently inhibited the interactions between NMDARs and PSD-95. Furthermore, behavior testing showed that animals given Tat-PDZ exhibited significantly reduced established inflammatory pain behaviors compared to vehicle-treated group. Our results indicate that by disrupting NMDAR/PSD-95 protein interactions, the Tat-PDZ cell-permeable fusion peptides provide a new approach for inflammatory pain therapy.

Abstract translation: 细胞通透性融合肽Tat-PDZ可以剂量依赖地降低麻醉阈值。 中枢神经系统突触中PDZ结构域介导的蛋白质相互作用在麻醉的分子机制中起着重要作用。 此外，Tat-PDZ细胞通透性融合肽在腹膜内注射后在细胞内递送到中枢神经系统的神经元。 通过体外和体内结合测定，我们发现Tat-PDZ剂量依赖性地抑制NMDAR和PSD-95之间的相互作用。 此外，行为测试表明，与载体治疗组相比，给予Tat-PDZ的动物表现出显着降低的建立的炎性疼痛行为。 我们的研究结果表明，通过破坏NMDAR / PSD-95蛋白相互作用，Tat-PDZ细胞可渗透融合肽为炎症性疼痛治疗提供了新的途径。

公开(公告)号：US20100204100A1

公开(公告)日：2010-08-12

申请号：US12596330

申请日：2008-04-17

Applicant: Roger A. Johns ,  Feng Tao

Inventor： Roger A. Johns ,  Feng Tao

IPC: A61K38/16 ,  C07K14/00 ,  A61P25/00 ,  A61P23/00

CPC classification number: C07K14/005 ,  A61K38/00 ,  C07K2319/10 ,  C07K2319/70 ,  C12N2740/16322

Abstract: Cell-permeant fusion peptides Tat-PDZ can dose-dependently reduce the threshold for anesthesia. PDZ domain-mediated protein interactions at synapses in the central nervous system play an important role in the molecular mechanisms of anesthesia. Moreover, Tat-PDZ cell-permeant fusion peptides are delivered intracellularly into neurons in the central nervous system subsequent to intraperitoneally injection. By in vitro and in vivo binding assays, we found that the Tat-PDZ dose-dependently inhibited the interactions between NMDARs and PSD-95. Furthermore, behavior testing showed that animals given Tat-PDZ exhibited significantly reduced established inflammatory pain behaviors compared to vehicle-treated group. Our results indicate that by disrupting NMDAR/PSD-95 protein interactions, the Tat-PDZ cell-permeable fusion peptides provide a new approach for inflammatory pain therapy.

Abstract translation: 细胞通透性融合肽Tat-PDZ可以剂量依赖地降低麻醉阈值。 中枢神经系统突触中PDZ结构域介导的蛋白质相互作用在麻醉的分子机制中起着重要作用。 此外，Tat-PDZ细胞通透性融合肽在腹膜内注射后在细胞内递送到中枢神经系统的神经元。 通过体外和体内结合测定，我们发现Tat-PDZ剂量依赖性地抑制NMDAR和PSD-95之间的相互作用。 此外，行为测试表明，与载体治疗组相比，给予Tat-PDZ的动物表现出显着降低的建立的炎性疼痛行为。 我们的研究结果表明，通过破坏NMDAR / PSD-95蛋白相互作用，Tat-PDZ细胞可渗透融合肽为炎症性疼痛治疗提供了新的途径。

公开(公告)号：US20110178160A1

公开(公告)日：2011-07-21

申请号：US13073238

申请日：2011-03-28

Applicant: Yuanxiang Tao ,  Roger A. Johns

Inventor： Yuanxiang Tao ,  Roger A. Johns

IPC: A61K31/7088 ,  A61P29/00 ,  C07H21/02

CPC classification number: C07K14/47 ,  A61K38/1709 ,  A61K45/06 ,  A63B59/50 ,  A63B2102/18 ,  A61K2300/00

Abstract: PSD-95/SAP90 antisense-treated animals not only experience a significant decrease in MAC for isoflurane, but also experience an attenuation in the NMDA-induced increase in isoflurane MAC. PSD-95/SAP90 appears to mediate the role of the NMDA receptor in determining the MAC of inhalational anesthetics. Suppression of the expression of PSD-95/SAP90 in the spinal cord significantly attenuates responses to painful stimuli mediated through the N-methyl-D-aspartate receptor activation. In spinal cord neurons PSD-95/SAP90 interacts with the N-methyl-D-aspartate receptor subunits 2A/2B. Activation of the N-methyl-D-aspartate receptor in spinal hyperalgesia results in association of the N-methyl-D-aspartate receptor with PSD-95/SAP90. PSD-95/SAP90 is required for hyperalgesia triggered via the N-methyl-D-aspartate receptor at the spinal cord level.

Abstract translation: PSD-95 / SAP90反义处理的动物不仅经历异氟烷的MAC显着降低，而且在NMDA诱导的异氟醚MAC增加中经历衰减。 PSD-95 / SAP90似乎介导NMDA受体在确定吸入麻醉剂的MAC中的作用。 抑制脊髓中PSD-95 / SAP90的表达显着减弱通过N-甲基-D-天冬氨酸受体激活介导的疼痛刺激的反应。 在脊髓神经元PSD-95 / SAP90与N-甲基-D-天冬氨酸受体亚基2A / 2B相互作用。 脊髓痛觉过敏中N-甲基-D-天冬氨酸受体的激活导致N-甲基-D-天冬氨酸受体与PSD-95 / SAP90的结合。 需要PSD-95 / SAP90用于在脊髓水平通过N-甲基-D-天冬氨酸受体触发的痛觉过敏。

公开(公告)号：US08343491B2

公开(公告)日：2013-01-01

申请号：US12518857

申请日：2007-12-18

Applicant: Roger A. Johns ,  Qingning Su ,  Hunter Clay Champion

Inventor： Roger A. Johns ,  Qingning Su ,  Hunter Clay Champion

IPC: A61K39/395

CPC classification number: G01N33/6884 ,  A61K2039/505 ,  C07K14/4702 ,  C07K16/18 ,  C07K16/22 ,  C07K16/40 ,  C12N9/1205 ,  C12Y207/10002 ,  G01N33/54306 ,  G01N33/74 ,  G01N2333/47 ,  G01N2333/4703 ,  G01N2333/912 ,  G01N2500/02 ,  G01N2800/12 ,  G01N2800/122 ,  G01N2800/7038

Abstract: Hypoxia induced mitogenic factor (HIMF) is a member of the “found in inflammatory zone” (FIZZ)/resistin family of proteins and has potent mitogenic, angiogenic, and vasoconstrictive effects in the lung vasculature. We use antibodies to HIMF to treat certain diseases including adult respiratory distress syndrome, radiation-induced pulmonary fibrosis, idiopathic pulmonary fibrosis, and emphysema.

Abstract translation: 缺氧诱导的促有丝分裂因子（HIMF）是炎症区（FIZZ）/抵抗素蛋白家族中发现的成员，并且在肺血管系统中具有有效的促有丝分裂，血管生成和血管收缩作用。 我们使用HIMF抗体来治疗某些疾病，包括成人呼吸窘迫综合征，放射性肺纤维化，特发性肺纤维化和肺气肿。

公开(公告)号：US06476007B2

公开(公告)日：2002-11-05

申请号：US09731876

申请日：2000-12-08

Applicant: Yuanxiang Tao ,  Roger A. Johns

Inventor： Yuanxiang Tao ,  Roger A. Johns

IPC: A61K31675

CPC classification number: A61K31/7076 ,  A61K31/00 ,  A61K31/52 ,  A61K31/538 ,  A61K45/06 ,  C12Q1/44 ,  C12Q1/485 ,  G01N2500/04 ,  Y10S514/816 ,  Y10S514/817 ,  Y10S514/818 ,  Y10S514/922

Abstract: Several lines of evidence have shown a role for the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling pathway in the development of spinal hyperalgesia. However, the roles of effectors for cGMP are not fully understood in the processing of pain in the spinal cord. cGMP-dependent protein kinase (PKG) I&agr; but not PKGI&bgr; was localized in the neuronal bodies and processes, and was distributed primarily in the superficial laminae of the spinal cord. Intrathecal administration of an inhibitor of PKGI&agr;, Rp-8-[(4-Chlorophenyl)thio]-cGMPS triethylamine, produces significant antinociception. Moreover, PKGI&agr; protein expression was dramatically increased in the lumbar spinal cord after noxious stimulation. This upregulation of PKGI&agr; expression was completely blocked not only by a neuronal NO synthase inhibitor, and a soluble guanylate cyclase inhibitor, but also by an N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801. Noxious stimulation not only initially activates but also later upregulates PKGI&agr; expression in the superficial laminae via an NMDA-NO-cGMP signaling pathway, suggesting that PKGI&agr; plays an important role in the central mechanism of inflammatory hyperalgesia in the spinal cord.

Abstract translation: 一些证据表明，一氧化氮（NO）/环鸟嘌呤一磷酸（cGMP）信号通路在脊髓痛觉过敏发展中起作用。 然而，在治疗脊髓疼痛方面，cGMP的作用者的作用尚未完全了解。 cGMP依赖性蛋白激酶（PKG），而不是PKGIbeta位于神经元体内和过程中，主要分布于脊髓浅层。 PKGIalpha，Rp-8 - [（4-氯苯基）硫代] -cGMPS三乙胺的抑制剂的鞘内施用产生显着的抗感受伤害。 此外，PKGIalpha蛋白表达在有害刺激后在腰脊髓中急剧增加。 PKGIalpha表达的上调不仅被神经元NO合成酶抑制剂和可溶性鸟苷酸环化酶抑制剂，而且被N-甲基-D-天冬氨酸（NMDA）受体拮抗剂MK-801完全阻断。 有毒刺激不仅最初激活，而且后来通过NMDA-NO-cGMP信号途径上调表达层中的PKGIalpha表达，表明PKGα在脊髓炎症痛觉过敏的中枢机制中起重要作用。

公开(公告)号：US07294476B2

公开(公告)日：2007-11-13

申请号：US11200169

申请日：2005-08-10

Applicant: Roger A. Johns ,  Yuanxiang Tao

Inventor： Roger A. Johns ,  Yuanxiang Tao

IPC: G01N33/567

CPC classification number: A61K31/7076 ,  A61K31/00 ,  A61K31/52 ,  A61K31/538 ,  A61K45/06 ,  C12Q1/44 ,  C12Q1/485 ,  G01N2500/04 ,  Y10S514/816 ,  Y10S514/817 ,  Y10S514/818 ,  Y10S514/922

Abstract: Several lines of evidence have shown a role for the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling pathway in the development of spinal hyperalgesia. However, the roles of effectors for cGMP are not fully understood in the processing of pain in the spinal cord. cGMP-dependent protein kinase (PKG) Iα but not PKGIβ was localized in the neuronal bodies and processes, and was distributed primarily in the superficial laminae of the spinal cord. Intrathecal administration of an inhibitor of PKGIα, Rp-8-[(4-Chlorophenyl)thio]-cGMPS triethylamine, produces significant antinociception. Moreover, PKGIα protein expression was dramatically increased in the lumbar spinal cord after noxious stimulation. This upregulation of PKGIα expression was completely blocked not only by a neuronal NO synthase inhibitor, and a soluble guanylate cyclase inhibitor, but also by an N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801. Noxious stimulation not only initially activates but also later upregulates PKGIα expression in the superficial laminae via an NMDA-No-cGMP signaling pathway, suggesting that PKGIα plays an important role in the central mechanism of inflammatory hyperalgesia in the spinal cord.

Abstract translation: 一些证据表明，一氧化氮（NO）/环鸟嘌呤一磷酸（cGMP）信号通路在脊髓痛觉过敏发展中起作用。 然而，在治疗脊髓疼痛方面，cGMP的作用者的作用尚未完全了解。 cGMP依赖性蛋白激酶（PKG），而不是PKGIbeta位于神经元体内和过程中，主要分布于脊髓浅层。 PKGIalpha，Rp-8 - [（4-氯苯基）硫代] -cGMPS三乙胺的抑制剂的鞘内施用产生显着的抗感受伤害。 此外，PKGIalpha蛋白表达在有害刺激后在腰脊髓中急剧增加。 PKGIalpha表达的上调不仅被神经元NO合成酶抑制剂和可溶性鸟苷酸环化酶抑制剂，而且被N-甲基-D-天冬氨酸（NMDA）受体拮抗剂MK-801完全阻断。 有毒刺激不仅最初激活，而且后来通过NMDA-No-cGMP信号通路上调表达薄层中的PKGIalpha表达，表明PKGα在脊髓炎症性痛觉过敏的中枢机制中起重要作用。

公开(公告)号：US08148347B2

公开(公告)日：2012-04-03

申请号：US13073238

申请日：2011-03-28

Applicant: Yuanxiang Tao ,  Roger A. Johns

Inventor： Yuanxiang Tao ,  Roger A. Johns

IPC: C12N15/11 ,  C07H21/04

CPC classification number: C07K14/47 ,  A61K38/1709 ,  A61K45/06 ,  A63B59/50 ,  A63B2102/18 ,  A61K2300/00

Abstract: PSD-95/SAP90 antisense-treated animals not only experience a significant decrease in MAC for isoflurane, but also experience an attenuation in the NMDA-induced increase in isoflurane MAC. PSD-95/SAP90 appears to mediate the role of the NMDA receptor in determining the MAC of inhalational anesthetics. Suppression of the expression of PSD-95/SAP90 in the spinal cord significantly attenuates responses to painful stimuli mediated through the N-methyl-D-aspartate receptor activation. In spinal cord neurons PSD-95/SAP90 interacts with the N-methyl-D-aspartate receptor subunits 2A/2B. Activation of the N-methyl-D-aspartate receptor in spinal hyperalgesia results in association of the N-methyl-D-aspartate receptor with PSD-95/SAP90. PSD-95/SAP90 is required for hyperalgesia triggered via the N-methyl-D-aspartate receptor at the spinal cord level.

Abstract translation: PSD-95 / SAP90反义处理的动物不仅经历异氟烷的MAC显着降低，而且在NMDA诱导的异氟醚MAC增加中经历衰减。 PSD-95 / SAP90似乎介导NMDA受体在确定吸入麻醉剂的MAC中的作用。 抑制脊髓中PSD-95 / SAP90的表达显着减弱通过N-甲基-D-天冬氨酸受体激活介导的疼痛刺激的反应。 在脊髓神经元PSD-95 / SAP90与N-甲基-D-天冬氨酸受体亚基2A / 2B相互作用。 脊髓痛觉过敏中N-甲基-D-天冬氨酸受体的激活导致N-甲基-D-天冬氨酸受体与PSD-95 / SAP90的结合。 需要PSD-95 / SAP90用于在脊髓水平通过N-甲基-D-天冬氨酸受体触发的痛觉过敏。

公开(公告)号：US20100028355A1

公开(公告)日：2010-02-04

申请号：US12518857

申请日：2007-12-18

Applicant: Roger A. Johns ,  Qingning Su ,  Hunter Clay Champion

Inventor： Roger A. Johns ,  Qingning Su ,  Hunter Clay Champion

IPC: A61K39/395 ,  A61K31/7052 ,  G01N33/53 ,  G01N33/68 ,  A61P11/00 ,  C12Q1/02 ,  A61P29/00

CPC classification number: G01N33/6884 ,  A61K2039/505 ,  C07K14/4702 ,  C07K16/18 ,  C07K16/22 ,  C07K16/40 ,  C12N9/1205 ,  C12Y207/10002 ,  G01N33/54306 ,  G01N33/74 ,  G01N2333/47 ,  G01N2333/4703 ,  G01N2333/912 ,  G01N2500/02 ,  G01N2800/12 ,  G01N2800/122 ,  G01N2800/7038

Abstract: Hypoxia induced mitogenic factor (HIMF) is a member of the “found in inflammatory zone” (FIZZ)/resistin family of proteins and has potent mitogenic, angiogenic, and vasoconstrictive effects in the lung vasculature. The receptor/binding partners for this family of proteins have been largely unknown. We identified Bruton's tyrosine kinase (BTK) as a functional HIMF binding partner through GST-HIMF pull-downs and mass spectrometry. Using primary cultured HIMF-stimulated murine bone marrow cells, we demonstrated that BTK was recruited to the leading edge of the cells. We also demonstrated that BTK and the closely related tyrosine kinase Fyn, colocalized at the growth cone process in these cells. HIMF stimulation induced BTK autophosphorylation, which peaked at 2.5 minutes. A transwell migration assay showed that treatment with recombinant murine HIMF induced migration of primary cultured bone marrow cells, which was completely blocked by the BTK inhibitor, LFM-A13. In vivo studies, using the rat hindlimb ischemia model, revealed that HIMF can stimulate angiogenesis in the hypoxic tissue probably through inducing the migration of endothelial progenitor cells (EPCs) to areas of active angiogenesis. Our results indicate that HIMF may acts as a chemotactic molecule in stimulating the migration of leukocytes/EPCs from bone marrow to targeted tissues through activation of the BTK pathway.

Abstract translation: 缺氧诱导的促有丝分裂因子（HIFF）是“发现在炎症区”（FIZZ）/抵抗素蛋白家族中的成员，并且在肺血管系统中具有有效的促有丝分裂，血管生成和血管收缩作用。 这种蛋白质家族的受体/结合配偶体在很大程度上是未知的。 我们通过GST-HIMF下拉和质谱鉴定了Bruton的酪氨酸激酶（BTK）作为功能性HIMF结合配偶体。 使用原代培养的HIMF刺激的鼠骨髓细胞，我们证明BTK被招募到细胞的前沿。 我们还表明，BTK和密切相关的酪氨酸激酶Fyn在这些细胞的生长锥过程中共定位。 HIMF刺激诱导BTK自磷酸化，其在2.5分钟达到峰值。 transwell迁移测定显示用重组鼠HIMF处理诱导原代培养的骨髓细胞的迁移，其被BTK抑制剂LFM-A13完全阻断。 使用大鼠后肢缺血模型的体内研究表明，HIMF可以通过诱导内皮祖细胞（EPCs）迁移到活动性血管生成区域来刺激缺氧组织中的血管生成。 我们的研究结果表明，HIMF可以作为趋化分子，通过激活BTK途径刺激白细胞/ EPCs从骨髓向目标组织的迁移。

公开(公告)号：US07030100B2

公开(公告)日：2006-04-18

申请号：US10183635

申请日：2002-06-28

Applicant: Yuanxiang Tao ,  Roger A. Johns

Inventor： Yuanxiang Tao ,  Roger A. Johns

IPC: A61K31/70 ,  A61K43/04

CPC classification number: A61K31/7076 ,  A61K31/00 ,  A61K31/52 ,  A61K31/538 ,  A61K45/06 ,  C12Q1/44 ,  C12Q1/485 ,  G01N2500/04 ,  Y10S514/816 ,  Y10S514/817 ,  Y10S514/818 ,  Y10S514/922

Abstract: Several lines of evidence have shown a role for the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling pathway in the development of spinal hyperalgesia. However, the roles of effectors for cGMP are not fully understood in the processing of pain in the spinal cord. cGMP-dependent protein kinase (PKG) Iα but not PKGIβ was localized in the neuronal bodies and processes, and was distributed primarily in the superficial laminae of the spinal cord. Intrathecal administration of an inhibitor of PKGIα, Rp-8-[(4-Chlorophenyl)thio]-cGMPS triethylamine, produces significant antinociception. Moreover, PKGIα protein expression was dramatically increased in the lumbar spinal cord after noxious stimulation. This upregulation of PKGIα expression was completely blocked not only by a neuronal NO synthase inhibitor, and a soluble guanylate cyclase inhibitor, but also by an N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801. Noxious stimulation not only initially activates but also later upregulates PKGIα expression in the superficial laminae via an NMDA-NO-cGMP signaling pathway, suggesting that PKGIα plays an important role in the central mechanism of inflammatory hyperalgesia in the spinal cord.