JP4754216B2 - dsRNA投与による神経疾患の処置 - Google Patents
dsRNA投与による神経疾患の処置 Download PDFInfo
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- JP4754216B2 JP4754216B2 JP2004533463A JP2004533463A JP4754216B2 JP 4754216 B2 JP4754216 B2 JP 4754216B2 JP 2004533463 A JP2004533463 A JP 2004533463A JP 2004533463 A JP2004533463 A JP 2004533463A JP 4754216 B2 JP4754216 B2 JP 4754216B2
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Description
本発明は神経疾患の処置方法、および慢性疼痛を処置するための医薬組成物に関する。
特に標的mRNAに相補的な短い一本鎖オリゴヌクレオチドまたはオリゴリボヌクレオチドまたは改変オリゴヌクレオチドにより、遺伝子の発現を阻害する方法は「アンチセンス」として知られている。遺伝子機能を解明する助けとなるツールとしてのアンチセンスオリゴヌクレオチド(ASO)の使用は十分記載されている。アンチセンスオリゴヌクレオチドはまた、多様な疾病の薬剤としても評価されている。
本発明は、神経疾患を処置または改善する方法であって、それを必要とする対象への有効量の二本鎖(ds)RNAの髄腔内注射を含み、該dsRNAが標的遺伝子の発現を阻害する方法に関する。好ましい一実施形態では、神経疾患はアルツハイマー病、パーキンソン病、多発性硬化症、統合失調症(schizophrenia)、癲癇、鬱病および疼痛からなる群から選択される。より好ましい一実施形態では、疾患は慢性疼痛、好ましくは慢性神経因性疼痛、癌性疼痛または骨関節炎性疼痛である。別の好ましい一実施形態では、疾患は異痛または痛覚過敏症である。あるいは、疾患は炎症性慢性疼痛である。別の好ましい一実施形態では、標的遺伝子はプリン受容体P1もしくはP2、ガラニンR1受容体、バニロイド受容体1、電位依存性カルシウムチャネル(N型)、テトロドトキシン抵抗性ナトリウムチャネルNav1.8(PN3/SNS)、TRPM8、IL-24、IL-20RαまたはIL-20Rβからなる群から選択される。特に好ましいものはP2受容体であり、最も好ましいものはP2X3またはP2X2である。さらに好ましい標的遺伝子としては、Mob-5またはMMP7がある。
本明細書に記載される発明は記載されている特定の方法論、プロトコール、および試薬に限定されず、可変であると考えられる。また、本明細書で用いる用語は単に特定の実施形態を記載するためのものであり、本発明の範囲を何ら限定するものではないと考えられる。
P2X3またはMOB-5をターゲッティングするオリゴヌクレオチドの合成
アンチセンスオリゴヌクレオチド(ASO)
P2X3およびGAPDHに対するASOは、2’-MOE基で改変した3’末端に9個のヌクレオチドを有する完全にホスホロチオエート化された18マーであり、ホスホルアミダイト化学(欧州特許出願EP992506 A2)を用いて合成し、HPLC精製し、エレクトロスプレー質量分析およびキャピラリーゲル電気泳動により同定されたものである。ミスマッチを含む対照オリゴヌクレオチドについては、マッチオリゴヌクレオチドのおよその塩基組成を維持するものであった(表1)。
本発明に記載される改変された合成オリゴリボヌクレオチドおよび改変されたアンチセンスオリゴヌクレオチドは、インビトロ(in vitro)用としてはABI394またはExpedite/Moss Synthesizers (Applied Biosystems)にて、また、in vivo用としてはOligoPilot II (Amersham Pharmacia Biotech)にて、標準的なホスホルアミダイト化学を用いて製造する。ホスホルアミダイトはアセトニトリルに0.05M濃度(Oligopilot IIでは0.2M)で溶かし、アセトニトリル中ベンジミダゾリウムトリフレートの0.2M溶液よりホスホルアミダイトを活性化させることでカップリングを行う。カップリング時間は通常3〜6分の間である。最初のキャッピングは標準的なキャッピング試薬を用いて行う。硫化はN-エチル、N-フェニル-5-アミノ-1,2,4-ジチアゾール-3-チオンの0.05M溶液を用いて2分間行う(EP-A-0992506に記載)。酸化はTHF/ピリジン/水(1:1:1)中0.1Mのヨウ素溶液により2分間行う。2回目のキャッピングは酸化または硫化の後に行う。次のカップリングのため、オリゴヌクレオチド成長鎖をジクロロメタンまたはジクロロエタン中2%のジクロロ酢酸により脱トリチル化する。配列が完成した後、支持体に結合した化合物を切断し、オリゴリボヌクレオチドに関しては35℃で6時間、メチルアミン溶液(41%水性メチルアミン/33%エタノール性メチルアミン1:1 v/v)による、また、アンチセンスオリゴヌクレオチドに関しては、55℃で16時間、32%アンモニア水溶液による「トリチル・オン(Trityl-on)」のようにして脱保護する。得られた懸濁液を凍結乾燥により乾固させる。オリゴリボヌクレオチドに関しては、1Mフッ化テトラブチルアンモニウムで50℃にて10分間、さらに35℃にて6時間処理すると、2’-O-シリル基が除去される。得られた粗溶液をそのままRP-HPLCで精製する。精製した脱トリチル化化合物をエレクトロスプレー質量分析およびキャピラリーゲル電気泳動で分析し、それらの吸光係数に従い、UV 260nMにより定量する。ラットP2X3およびMOB-5に対するオリゴリボヌクレオチドおよびアンチセンスオリゴヌクレオチド、ならびにそれらの対照をそれぞれ表3および4に示す。
チャイニーズハムスター卵巣細胞(CHO-K1, ATCC CCL61)を、ベクターpRK7(John Woodから厚意により入手; Chen, C.C et al. (1995). Nature 377, 428-430)中の完全ラットP2X3 cDNA配列で安定的にトランスフェクトした。トランスフェク細胞の選択を可能とするため、ベクターを10倍過剰の、ネオマイシン耐性遺伝子を含むpMC1neo (Stratagene)で同時エレクトロポレーションした。細胞を、10%(v/v)ウシ胎児血清(FBS)、2mMグルタミン、および10.000IU/500mlペニシリン/ストレプトマイシンを添加したMinimal Essential Mediumα(MEMα)で培養した。
rP2X3インサートを、鋳型としてラットDRG RNA由来のRTを用い、以下に挙げたオリゴを用いてPCRにより得た。
P2X3-Hind-F: CGCAAGCTTGGCTGTGAGCAGTTTCTCAGTATGAACTTG(配列番号17)
P2X3- SacI-R: CTTGAGCTCGGGAAGAGGCCCTAGTGACCAATAG(配列番号18)
注:下線の配列はP2X3相補的オリゴに付加されたHindIII制限酵素部位である。リバースプライマーにおけるSacI部位はクローニングには用いなかった。
用いた細胞系統はRBA (ATCC番号1747)であった。これは、ラット皮膚に由来し、平板状に極めて強固に結合して増殖し、皮膚様の形態で、本来rasおよびmob-5(rasの下流にあることが知られている)を発現する。
標準的な電気トランスフェクション(Bioradキュベット0.4cm中、106細胞/125μl、250V、0.3μF、無限抵抗)を用い、CHO-rP2X2/3細胞を0.15、0.3、0.6または1.2ナノモルのASOまたはsiRNA二重らせんでトランスフェクトした。エレクトロポレーション後、すぐ、サンプルを6mlの培養培地と合わせた。結果として、ASOまたはsiRNA試薬の対応する最終濃度は10、50、100または200nMとなった。細胞を非被覆96ウェルプレート(Costar, カタログ番号#3904)にプレーティングし、37℃で24時間または48時間インキュベートした後、それぞれRNAまたはタンパク質を抽出した。
RNeasy96キット(Qiagen)を用い、全RNAを抽出および精製した。リアルタイムPCRのためのプライマー対およびFAM標識TaqManプローブはPrimer Express v2.0プログラム(ABI PRISM, PE Biosystems)を用いてデザインした。Q-PCR反応のためには、TaqMan PCR試薬キットプロトコール(Eurogentec)に従い、50ngの全RNAを、全量25μl中、5’および3’プライマー(各10μM)、TaqManプローブ(5μM)、MuLV逆転写酵素6.25u, PE Biosystems)、RNアーゼOut RNアーゼ阻害剤(10u, Life Technologies)およびTaqMan PCR試薬キット(Eurogentec)の成分と混合した。逆転写およびリアルタイムPCRを下記のようにGeneAmp Sequence Detector 5700 (PE Biosystems)にて行った:50℃で2分の逆転写、95℃で10分の変性、その後、95℃で15秒の変性および60℃で1分のアニーリングおよび伸張を50サイクル。遺伝子発現の相対数値はABI PRISM 7700ユーザー会報#2 (PE Biosystems)に記載のようにして算出した。
6ウェルプレートで増殖させた細胞をPBSで洗浄し、141mM NaCl、5mM KCl、2.5mM Tris pH7.4、50nM Va3VO4、0.1%(v/v)Nonidet P-40(100%)、および0.06gプロテアーゼ阻害剤/100mlを含むバッファーで溶解させた。溶解物を14000rpmで10分遠心分離した。上清中の可溶化タンパク質に対し、NOVEX(商標)Mini-CellシステムにてNuPAGE(商標)4〜12% Bis-TrisゲルでSDSポリアクリルアミドゲル電気泳動を行った後、PVDFメンブラン(Millipore)に移した。これらのフィルターを、ECFウエスタンブロッティングキット(Amersham Pharmacia Biotech)に含まれているブロッキングバッファーで1時間ブロッキングし、0.05% Tween 20を含む1×PBS, pH7.4中で数回洗浄し、1:5000希釈の一次抗P2X3抗体(Neuromicsから購入)とともに1時間インキュベートした。その後、これらのフィルターを、間に数回洗浄しながら、製造業者の指示に従い、ECFウエスタンブロッティングキットの二次抗体、三次抗体およびECF基質とともにインキュベートした。可視化されたバンドの定量はソフトウエアImageQuant(商標)(Molecular Dynamics)により行った。
FLIPR実験は次のようにして行った。要するに、2.5mMプロベニシドの存在下で30〜45分間細胞にfluo-4 AMを加え、HBSS(Gibco)+20mM HEPESで2回洗浄し、蛍光リーダー(FLIPR, Molecular Devices)に移した。薬物のプレートは5倍の最終濃度で調製した。Fluo-4の蛍光は0.5Hzで3分測定した。20ポイントのベースラインが検出されたところでアゴニストを添加した。
慢性神経因性疼痛のインビボ(in vivo)動物モデルには次のものが含まれる。
Seltzerモデル
Seltzerモデル(Seltzer et al. (1990) Pain 43: 205-218)では、ラットを麻酔し、一方の大腿(通常は左)の正中を上方に小さく切開し、坐骨神経を露出させる。後大腿二頭筋半腱様筋神経が総坐骨神経を分岐する点から少し離れた転子付近の部位で、この神経の周囲の結合組織を注意深く取り除く。7-0縫合糸を、3/8カーブのリバースカッティングミニニードル(reversed-cutting mini-needle)で神経に挿入し、神経の厚みの背側1/3〜1/2が結紮内に保持されるようにしっかり結紮する。縫合糸とクリップで筋肉と皮膚を閉じ、傷口に抗生物質粉末を塗布する。擬似手術動物では、座骨神経を露出させるが、結紮はせず、擬似手術動物の場合と同様、傷口を閉じる。
CCIモデル(Bennett, G.J. and Xie, Y.K. Pain (1988) 33: 87-107)では、ラットを麻酔し、一方の大腿(通常は左)の正中を上方に小さく切開し、坐骨神経を露出させる。この神経の周囲の結合組織を取り除き、この神経を約1mm間隔で4箇所、4/0腸線縫合糸で、神経の表面がわずかにくびれるように軽く結紮する。上記のように縫合糸とクリップで傷口を閉じる。擬似手術動物では、座骨神経を露出させるが、結紮はせず、擬似手術動物の場合と同様、傷口を閉じる。
末梢神経に対する損傷を含むSeltzerおよびCCIモデルに対し、Chungモデルは脊髄神経の結紮を含む(Kim, S.O. and Chung, J.M. Pain (1992): 50:355-363)。このモデルでは、ラットを麻酔し、腹臥位に置き、脊髄の左側をL4-S2レベルで切開する。坐骨神経は分岐してL4、L5およびL6脊髄神経を形成しているが、傍脊柱筋を深く切り、L4-S2レベルで脊髄突起から筋肉を分離すると坐骨神経の一部が現れる。L6横突起を小さな骨鉗子で注意深く取り除くと、これらの脊髄神経が見える。L5脊髄神経を単離し、7-0縫合糸でしっかり結紮する。傷口を一重の筋縫合糸(6-0シルク)および1または2個の皮膚縫合クリップで閉じ、抗生物質粉末を塗布する。擬似手術動物では、L5神経を露出させるが、結紮はせず、上記と同様に傷口を閉じる。
軸索切断モデルは坐骨神経の完全切断と結紮を含む。これらの神経終末は神経小丘を形成するが、この神経は再生できないので、このモデルでは挙動相関が見られず、脚は恒常的に麻痺する(Kingery and Vallin, Pain 38, 321-32, 1989)。
このモデルでは、坐骨神経を腸骨弓の領域で穿孔する。神経に対する顕在的な損傷は見られないが、局部的な腫脹が、神経が腸骨弓の下を通っているので、神経に対する圧力を高める。このモデルは腸骨でしばしば見られる症状を模倣したものである。
フロイントの完全アジュバントにより誘発される機械的痛覚過敏症を慢性炎症性疼痛のモデルとして用いることができる(Stein, C. et al. Pharmacol. Biochem. Behav. (1988) 31 :445-451)。このモデルでは、典型的には、雄のSprague-DawleyまたはWistarラット(200〜250g)の片方の後脚に25μlのフロイントの完全アジュバントを足底内注射する。この後脚に著しい炎症が起こる。一般に、炎症発生から24時間後、機械的痛覚過敏症が十分確立されたとみなされた際に薬物を投与して効力を評価する。
全ての慢性疼痛モデル(炎症性および神経因性)で、機械的痛覚過敏症は、Analgesymeter (Ugo-Basile, Milan)を用いて段階的に高まる圧力刺激に対する両後脚の引っ込め閾値(withdrawal threshold)を測定することにより評価する。機械的異痛は、両後脚の足底表面にvon Frey hairで適用する無害の機械的刺激に対する引っ込め閾値を測定することにより評価する。熱的痛覚過敏症は、各後脚の裏面に適用する無害の熱刺激に対する引っ込め閾値を測定することにより評価する。全てのモデルで、機械的痛覚過敏症および異痛ならびに熱的痛覚過敏症は外科術後1〜3日以内に発症し、少なくとも50日間持続する。本明細書に記載のアッセイでは、外科術前後に薬物を適用し、特に外科術後約14日目に痛覚過敏症の発症に対する効果を評価し、確立された痛覚過敏症を反転させるそれらの能力を決定することができる。
動物モデルに用いたオリゴヌクレオチド試薬には次のような名称が付けられている:ASO: NAS-6798(配列番号5)、MSO: NAS-6799(配列番号6)、P2X3 RNAi: NAS-8646およびNAS-8647(それぞれ、配列番号7および8)、P2X3 RNAiミスセンス: NAS-10104およびNAS-10105(それぞれ、配列番号19および20)。
dsRNAをバッファー(100mM KA 2mM MgAc、0.1749g HEPES遊離酸(M=238.3)、0.2102g NaCl/100ml RNアーゼフリー水; KOHで20℃にてpH 7.63)中、留置カニューレを通じて髄腔内投与した。ラットを麻酔し、正中のすぐ側部、腹側腸骨棘より尾側へ約10mmの背中の皮膚を切開した。無菌カテーテル(ポリエチレンPE10チューブ)をガイドカニューレ(20ゲージニードル)を通じて挿入し、髄腔内を頭部側へ3cmほぼL1レベルまで前進させた。次に、このカテーテルを、左または右側腹部に皮下挿入し、P2X3受容体またはMOB-5受容体 siRNA、ミスセンスsiRNAまたは生理食塩水(1μl/時、7日)を送達するオスモティックミニポンプ(Alzet)に接続した。創傷クリップで切開部を閉じ、抗生物質粉末を塗布した。実験から180ならびに220μg/日であれば毒性の徴候がないものと判定された。機械的痛覚過敏症は、α,β-メチレン-ATP(Me-ATP)の投与前は1日目と6日目、そしてMe-ATP投与1時間後に、Analgesymeter (Ugo-Basile, Milan)を用いて、高まる圧力刺激に対する両後脚の引っ込め閾値を測定することで評価した。限界値を250gに設定し、終点を引っ込め、鳴き声、または明らかな身もだえとした。各動物を無作為の順番で、一度だけ試験した。異なる試験動物群から得られた機械的痛覚過敏症データの統計学的有意性はANOVAの後にTukeyのHSD検定を用いて分析した。試験の最終日には、1.0μmol(10μl)のMe-ATPを対側の後脚に足底内(ipl)投与した。投与から10人看護、機械的痛覚過敏症に対する脚の引っ込め閾値を測定した。
ラットP2X3を発現する細胞系統へのsiRNAのトランスフェクション
アゴニスト誘発性の疼痛を有するラットにおける痛覚過敏症に対するP2X3 siRNAの効果
アゴニスト誘発性疼痛を有するラットにおける異痛に対するP2X3 siRNAの効果
神経因性疼痛を有するラットにおける機械的痛覚過敏症に対するP2X3 siRNAの効果(Seltzerモデル)
神経因性疼痛を有するラットにおける機械的異痛に対するP2X3 siRNAの効果(Seltzerモデル)
神経因性疼痛を有するラットにおける機械的痛覚過敏症に対するMOB-5 siRNAの効果(Seltzer モデル)
Claims (12)
- 有効量の二本鎖(ds)RNAを含み、該dsRNAがP2X3、P2X2またはMob-5の発現を阻害し、該dsRNAが過分極活性化型陽イオン非選択的(HCN)イオンチャンネル遺伝子に特異的ではなく、そして該dsRNAが:
(a)一方の鎖が配列番号7の配列を含み、他方の鎖が配列番号8の鎖を含むか、
(b)一方の鎖が配列番号9の配列を含み、他方の鎖が配列番号10の鎖を含むか、
(c)一方の鎖が配列番号13の配列を含み、他方の鎖が配列番号14の鎖を含むか、
(d)一方の鎖が配列番号15の配列を含み、他方の鎖が配列番号16の鎖を含むか、あるいは
(e)一方の鎖が配列番号21の配列を含み、他方の鎖が配列番号22の鎖を含む、
dsRNAの群から選択される、髄腔内注射によりそれを必要とする対象における疼痛を処置または改善するための薬剤。 - 該疼痛が慢性神経因性疼痛である、請求項1に記載の薬剤。
- 該疼痛が癌性疼痛、骨関節炎性疼痛、異痛および痛覚過敏症からなる群から選択される、請求項1に記載の薬剤。
- それを必要とする対象がヒトである、請求項1に記載の薬剤。
- それを必要とする対象がラットである、請求項1に記載の薬剤。
- 少なくとも200μgのdsRNAが髄腔内注射される、請求項1〜5のいずれか一項に記載の薬剤。
- dsRNAが15〜25ntの二本鎖領域を含む、請求項1〜6のいずれか一項に記載の薬剤。
- dsRNAが少なくとも1つのヌクレオチドの、アンチセンスもしくはセンス鎖、または両鎖上に3’オーバーハングを含む、請求項1〜7のいずれか一項に記載の薬剤。
- オーバーハングが少なくとも1つの改変ヌクレオチドを含む、請求項8に記載の薬剤。
- オーバーハングが少なくとも1つの2’−MOE改変ヌクレオチドを含む、請求項8に記載の薬剤。
- オーバーハングが4ウラシルを含む、請求項8に記載の薬剤。
- dsRNAが少なくとも1つのホスホロチオエート結合を含む、請求項1〜11のいずれか一項に記載の薬剤。
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US8742091B2 (en) * | 2001-06-20 | 2014-06-03 | Dainippon Sumitomo Pharma Co., Ltd. | Method of promoting nucleic acid transfer |
WO2004022075A1 (en) | 2002-09-04 | 2004-03-18 | Novartis Ag | Treatment of neurological disorders by dsrna adminitration |
RU2410430C2 (ru) * | 2004-08-31 | 2011-01-27 | Силентис С.А.У. | Способы и композиции для ингибирования экспрессии рецептора p2х7 |
SG158921A1 (en) * | 2004-10-27 | 2010-02-26 | Schering Corp | Compositions and methods for short interfering nucleic acid inhibition of nav1.8 |
JPWO2007020935A1 (ja) * | 2005-08-17 | 2009-02-26 | 小野薬品工業株式会社 | P2y12受容体および/またはp2y14受容体ブロッカーを含有してなる疼痛治療剤 |
WO2010036822A1 (en) * | 2008-09-24 | 2010-04-01 | Hollis-Eden Pharmaceuticals, Inc. | Patient populations and treatment methods |
AU2011314074B2 (en) * | 2010-09-28 | 2015-11-12 | Nono Inc. | ND2 peptides and methods of treating neurological disease |
US9458464B2 (en) | 2014-06-23 | 2016-10-04 | The Johns Hopkins University | Treatment of neuropathic pain |
WO2020037030A1 (en) * | 2018-08-14 | 2020-02-20 | University Of Maryland, Baltimore | Methods for targeting pain directed at metabolic pathways |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11504011A (ja) * | 1995-04-18 | 1999-04-06 | イナファーマ,インコーポレイテッド | 疼痛の治療方法 |
JP2002507191A (ja) * | 1997-04-28 | 2002-03-05 | サイトセラピューティクス,インコーポレイテッド | 痛覚脱失の生成または神経保護のためのコナントキンの使用 |
JP2005536438A (ja) * | 2001-06-08 | 2005-12-02 | オーソ−マクニール・フアーマシユーチカル・インコーポレーテツド | 過分極活性化型環状ヌクレオチド依存性チャンネルに標的を定めることによる疼痛の治療 |
Family Cites Families (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
IE72103B1 (en) * | 1987-08-12 | 1997-03-12 | Hem Res Inc | Promotion of host defense by systemic dsRNA treatment |
US5155689A (en) * | 1991-01-17 | 1992-10-13 | By-Word Technologies, Inc. | Vehicle locating and communicating method and apparatus |
US5214281A (en) * | 1992-03-20 | 1993-05-25 | Rowe Douglas J | Method for locating sub-terranean geological aggregate deposits |
US5724243A (en) * | 1995-02-10 | 1998-03-03 | Highwaymaster Communications, Inc. | Method and apparatus for determining expected time of arrival |
JP3371605B2 (ja) * | 1995-04-19 | 2003-01-27 | 日産自動車株式会社 | 大気効果表示機能付き鳥瞰図表示ナビゲーションシステム |
US5751450A (en) * | 1996-05-22 | 1998-05-12 | Medar, Inc. | Method and system for measuring color difference |
US6990458B2 (en) * | 1997-08-28 | 2006-01-24 | Csg Systems, Inc. | System and method for computer-aided technician dispatch and communication |
US6119124A (en) | 1998-03-26 | 2000-09-12 | Digital Equipment Corporation | Method for clustering closely resembling data objects |
US6240409B1 (en) | 1998-07-31 | 2001-05-29 | The Regents Of The University Of California | Method and apparatus for detecting and summarizing document similarity within large document sets |
ES2205667T3 (es) | 1998-10-08 | 2004-05-01 | Novartis Ag | Procedimiento para la sulfuracion de compuestos que contienen fosforo. |
DE19956568A1 (de) | 1999-01-30 | 2000-08-17 | Roland Kreutzer | Verfahren und Medikament zur Hemmung der Expression eines vorgegebenen Gens |
US7415313B2 (en) * | 2000-07-07 | 2008-08-19 | New Vectors Llc | Spatial coordination system |
US20030190635A1 (en) * | 2002-02-20 | 2003-10-09 | Mcswiggen James A. | RNA interference mediated treatment of Alzheimer's disease using short interfering RNA |
US6658423B1 (en) | 2001-01-24 | 2003-12-02 | Google, Inc. | Detecting duplicate and near-duplicate files |
AU2002306556A1 (en) * | 2001-02-22 | 2002-09-12 | University Of Maryland, Baltimore | Novel treatment of neurodegenerative diseases by altering levels of trkb isoforms and/or trkc isoforms |
US6845394B2 (en) * | 2001-04-16 | 2005-01-18 | Sun Microsystems, Inc. | Software delivery method with enhanced batch redistribution for use in a distributed computer network |
US20030008048A1 (en) | 2001-06-08 | 2003-01-09 | David Winston | Methods and compositions for helping the body resist the effects of the aging process |
EP1492516A2 (en) * | 2002-04-03 | 2005-01-05 | Novartis AG | Use of mob-5 in pain |
WO2004022075A1 (en) | 2002-09-04 | 2004-03-18 | Novartis Ag | Treatment of neurological disorders by dsrna adminitration |
BRPI0407107A (pt) | 2003-01-31 | 2006-01-24 | Novartis Ag | Infra-regulação de gene alvo com complexos de oligoribonucleotìdeo de filamento único e polìmero de polietilenoimina (pei) |
US20050132197A1 (en) | 2003-05-15 | 2005-06-16 | Art Medlar | Method and apparatus for a character-based comparison of documents |
US20070085716A1 (en) | 2005-09-30 | 2007-04-19 | International Business Machines Corporation | System and method for detecting matches of small edit distance |
US7499869B2 (en) * | 2006-02-02 | 2009-03-03 | Matthew Iknoian | System and method for scheduling employee shifts |
US7562186B2 (en) | 2006-04-11 | 2009-07-14 | Data Domain, Inc. | Efficient data storage using resemblance of data segments |
-
2003
- 2003-09-03 WO PCT/EP2003/009787 patent/WO2004022075A1/en active Application Filing
- 2003-09-03 US US10/525,312 patent/US20060030534A1/en not_active Abandoned
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-
2011
- 2011-02-21 JP JP2011034713A patent/JP2011178783A/ja active Pending
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH11504011A (ja) * | 1995-04-18 | 1999-04-06 | イナファーマ,インコーポレイテッド | 疼痛の治療方法 |
JP2002507191A (ja) * | 1997-04-28 | 2002-03-05 | サイトセラピューティクス,インコーポレイテッド | 痛覚脱失の生成または神経保護のためのコナントキンの使用 |
JP2005536438A (ja) * | 2001-06-08 | 2005-12-02 | オーソ−マクニール・フアーマシユーチカル・インコーポレーテツド | 過分極活性化型環状ヌクレオチド依存性チャンネルに標的を定めることによる疼痛の治療 |
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