JP6415570B2 - インターフェロン受容体の密度の減少に関連する疾患を予防又は治療するための方法 - Google Patents
インターフェロン受容体の密度の減少に関連する疾患を予防又は治療するための方法 Download PDFInfo
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Description
連続細胞系A-549を、100ng/mLの濃度のグルタリルヒスタミンで処理した。処理後16時間に、インターフェロン受容体のメッセンジャーRNAコピーの量を、リアルタイムPCR解析によって決定した。RNeasy Kit(Quiagen社)を使用することによって、RNAを単離した。RNA調製物をDNAase(RNase-free DNaseI(Ambion社))で処理した。Thermoscript RT-PCR System(Invitrogen社)を使用することによって、逆転写を行った。プライマー:
Ifnar1 (フォワード) 5' CACTGACTGTATATTGTGTGAAAGCCAGAG 3', (リバース) 5' CATCTATACTGGAAGAAGGTTTAAGTGATG 3' ;
Ifnar2 (フォワード) 5' ATTTCCGGTCCATCTTATCAT 3', (リバース) 5' ACTGAACAACGTTGTGTTCC 3'
を用いて、定量的PCRを実施した。
96-ウェルプレート中、ヒトマクロファージの10日初代培養物を、濃度:0.1、1.0、10若しくは100ng/mLのグルタリルヒスタミンの存在下又は非存在下で、24時間インキュベートした。次いで、細胞をパラホルムアルデヒド溶液で固定し(透過処理なし)、固定した細胞を、洗浄剤を含有する緩衝溶液(PBS-Tween)ですすぎ、ウシ血清アルブミン溶液でブロックし、インターフェロン受容体サブユニットIFNAR1又はIFNAR2に対する抗体を用いてインキュベートした。調製物の視覚化は、ホースラディシュペルオキシダーゼで標識された二次抗体で染色し、基質を加えることによって行った。ELISA分光光度計を使用して、光学密度を測定した。
細胞表面上のインターフェロン受容体の密度が増加すると、そのシグナル伝達がウイルス感染によって抑制される条件下で、内因性インターフェロンの弱いシグナルに対する細胞の感受性が高まらなければならない。インターフェロンシグナルに対する細胞の感作の影響を実証するために、細胞A-549を、100ng/mLの濃度のグルタリルヒスタミンで処理し、8時間又は24時間インキュベートした後、1IUインターフェロン-α(IFNα: Roferon-A3(登録商標), Roche Molecular Biochemicals社, Mannheim)の存在下又は非存在下で、細胞を溶解させた。細胞溶解物に、タンパク質MxA(sc-50509, Santa Cruz Biotechnology社)に対するモノクローナル抗体を使用することによって、免疫ブロット分析(ウエスタンブロット法)の手法を行い、基質(化学発光法の試薬であるSuper Signal West Femto Chemiluminescent Substrate(Thermo Fisher Scientific社))を使用する化学発光法によって、膜を染色した。細胞がインターフェロンで処理される際にインターフェロン刺激遺伝子(ISG)の発現を示す指標生成物として、タンパク質MxAを選択した。
マウスを単純ヘルペスウイルスHSV-1/CL及びHSV-2/VNに、10LD50を含む30μlの用量で脳内感染させた。
本発明に従って使用されるグルタリルヒスタミンの投与形態を、標準的な方法、例えば、混合、造粒、丸薬形成、溶解及び凍結乾燥のプロセス等によって調製する。
錠剤形態を、以下の成分を使用することによって調製する。
坐剤の製剤例
注射用溶液の製剤例:
Claims (18)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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RU2013140758 | 2013-09-03 | ||
RU2013140758/15A RU2595862C2 (ru) | 2013-09-03 | 2013-09-03 | Способ профилактики или лечения заболеваний, связанных с пониженной плотностью интерфероновых рецепторов |
PCT/RU2014/000656 WO2015034400A1 (ru) | 2013-09-03 | 2014-09-01 | Способ профилактики или лечения заболеваний, связанных с пониженной плотностью интерфероновых рецепторов |
Publications (2)
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JP2016529323A JP2016529323A (ja) | 2016-09-23 |
JP6415570B2 true JP6415570B2 (ja) | 2018-10-31 |
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JP2016540844A Expired - Fee Related JP6415570B2 (ja) | 2013-09-03 | 2014-09-01 | インターフェロン受容体の密度の減少に関連する疾患を予防又は治療するための方法 |
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US (1) | US9730915B2 (ja) |
EP (1) | EP3042664A4 (ja) |
JP (1) | JP6415570B2 (ja) |
KR (1) | KR20160048963A (ja) |
CN (1) | CN105555292B (ja) |
CA (1) | CA2943318A1 (ja) |
EA (1) | EA035359B1 (ja) |
HK (1) | HK1223299A1 (ja) |
IL (1) | IL244207B (ja) |
MX (1) | MX2016002856A (ja) |
RU (1) | RU2595862C2 (ja) |
SG (1) | SG11201601241YA (ja) |
UA (1) | UA121022C2 (ja) |
WO (1) | WO2015034400A1 (ja) |
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US11975002B2 (en) | 2016-03-04 | 2024-05-07 | Taiho Pharmaceutical Co., Ltd. | Preparation and composition for treatment of malignant tumors |
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EP0294160A1 (en) * | 1987-06-02 | 1988-12-07 | Schering Corporation | Treatment of chronic type b hepatitis with a combination of recombinant human alpha and gamma interferons |
RU2141483C1 (ru) * | 1997-07-04 | 1999-11-20 | Небольсин Владимир Евгеньевич | Производные пептидов или их фармацевтически приемлемые соли, способ их получения, применение и фармацевтическая композиция |
ES2381266T3 (es) * | 2002-07-15 | 2012-05-24 | Gilead Sciences, Inc. | Terapias de combinación de L-FMAU para el tratamiento de la infección por el virus de la hepatitis B |
US9504673B2 (en) * | 2009-05-21 | 2016-11-29 | LTD “Valenta-Intellekt” | Agent for the prophylaxis and treatment of highly pathogenic infectious diseases |
RU2438694C1 (ru) * | 2010-12-23 | 2012-01-10 | Общество с ограниченной ответственностью "ЦитоНИР" (ООО "ЦитоНИР") | Фармацевтическая композиция для лечения вирусных заболеваний |
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- 2014-09-01 WO PCT/RU2014/000656 patent/WO2015034400A1/ru active Application Filing
- 2014-09-01 CN CN201480048260.4A patent/CN105555292B/zh active Active
- 2014-09-01 EP EP14841881.7A patent/EP3042664A4/en not_active Withdrawn
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Also Published As
Publication number | Publication date |
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US9730915B2 (en) | 2017-08-15 |
IL244207B (en) | 2019-05-30 |
KR20160048963A (ko) | 2016-05-04 |
EP3042664A4 (en) | 2017-05-17 |
EA201600227A1 (ru) | 2016-06-30 |
RU2595862C2 (ru) | 2016-08-27 |
SG11201601241YA (en) | 2016-03-30 |
CN105555292B (zh) | 2020-01-10 |
HK1223299A1 (zh) | 2017-07-28 |
US20160354345A1 (en) | 2016-12-08 |
CA2943318A1 (en) | 2015-03-12 |
EA035359B1 (ru) | 2020-06-02 |
MX2016002856A (es) | 2016-06-22 |
CN105555292A (zh) | 2016-05-04 |
UA121022C2 (uk) | 2020-03-25 |
IL244207A0 (en) | 2016-04-21 |
JP2016529323A (ja) | 2016-09-23 |
WO2015034400A1 (ru) | 2015-03-12 |
EP3042664A1 (en) | 2016-07-13 |
RU2013140758A (ru) | 2015-03-10 |
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