JP5758381B2 - siRNA接合体及びその製造方法 - Google Patents
siRNA接合体及びその製造方法 Download PDFInfo
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Description
A−X−R−Y−B
(上記で、A及びBはそれぞれ独立に、親水性高分子または疎水性高分子化合物、X、Yはそれぞれ独立に、リンカーが媒介されていない共有結合またはリンカーが媒介された共有結合であり、RはsiRNAを示す。)
A−X−R
(上記で、Aは疎水性高分子化合物であり、Xはリンカーが媒介されていない共有結合またはリンカーが媒介された共有結合であり、RはsiRNAを示す。)
(上記で、Rは、アルキル、アルケニル、アルキニル、アリール、アリールアルキル、ヘテロアルキル、またはヘテロアリールであり;mは、2〜18の整数であり、nは5〜120の整数であり、Xは、水素、4−モノメトキシトリチル基、4,4’−ジメトキシトリチル基、4,4’,4”−トリメトキシトリチル基である。)
1)CPGに3−アミノプロピルトリエトキシシランを反応させて、LCAA−CPG(Long Chain Alkyl Amine Controlled Pore Glass)を形成する段階と、2)ポリエチレングリコールに4,4’−ジメトキシトリチルクロライドを反応させて、2−[ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)]を形成する段階と、3)前記2)で形成された化合物と下記化学式1の化合物とを反応させて、下記構造式Iで表される化合物を形成する段階と、4)前記形成された下記構造式Iの化合物とクロロギ酸4−ニトロフェニルとを反応させて、下記構造式II表される化合物を形成する段階と、5)前記3)で形成された下記構造式Iの化合物とN−スクシンイミジルトリフルオロ酢酸とを反応させて、下記構造式IIIで表される化合物を形成する段階と、6)前記1)で形成されたLCAA−CPG化合物を、前記3)から5)でそれぞれ形成された下記構造式I、IIまたはIIIの化合物とそれぞれ反応させる段階と、を含む前記第15に記載の3’−PEG−CPGの製造方法を提供するものである。
(上記で、Rは、アルキル、アルケニル、アルキニル、アリール、アリールアルキル、ヘテロアルキル、またはヘテロアリールであり;nは5以上120以下の整数である。)
A−X−R−Y−B
上記で、A及びBはそれぞれ独立に、親水性高分子または疎水性高分子化合物であり、X、Yはそれぞれ独立に、リンカーが媒介されていない共有結合またはリンカーが媒介された共有結合であり、RはsiRNAを示す。
A−X−R
上記で、Aは疎水性高分子化合物であり、Xはリンカーが媒介されていない共有結合またはリンカーが媒介された共有結合であり、RはsiRNAを示す。
上記で、固形支持体(solid support)は、例えば、CPG、ポリスチレン、シリカゲル、セルロース紙などを含むが、これに制限されず、Rは、アルキル、アルケニル、アルキニル、アリール、アリールアルキル、ヘテロアルキル、またはヘテロアリールであり;mは2〜18の整数であり、nは5〜120(Molar mass282〜5300)までの整数であり;Xは、4−モノメトキシトリチル基、4,4’−ジメトキシトリチル基、または4,4’,4”−トリメトキシトリチル基であって、酸で処理された後除去されて水素となる。固形支持体がCPGである場合、直径は40〜180μmであることが好ましく、500Å〜3000Åの空隙サイズを有することが好ましい。
2)ポリエチレングリコールに4,4’−ジメトキシトリチルクロライドを反応させて、2−[ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)]を形成する段階と、
3)前記2)で形成された化合物と下記化学式1の化合物とを反応させて、下記構造式Iで表される化合物を形成する段階と、
4)前記形成された下記構造式Iの化合物とクロロギ酸4−ニトロフェニルとを反応させて、下記構造式IIで表される化合物を形成する段階と、
5)前記3)で形成された下記構造式Iの化合物とN−スクシンイミジルトリフルオロ酢酸とを反応させて、下記構造式IIIで表される化合物を形成する段階と、
6)前記1)で形成されたLCAA−CPG化合物を、前記3)〜5)でそれぞれ形成された下記構造式I、IIまたはIIIの化合物とそれぞれ反応させる段階と、を含む下記構造式IVの3’−PEG−CPGの製造方法を提供する。
(上記で、Rは、アルキル、アルケニル、アルキニル、アリール、アリールアルキル、ヘテロアルキル、またはヘテロアリールであり;nは5以上120以下の整数である。)
1)前記ポリエチレングリコールが結合された固形支持体を利用して、標的遺伝子に対するsiRNAを製造する段階と、
2)siRNAの末端基とポリエチレングリコールとを共有結合で連結する段階と、を含む前記siRNA接合体の製造方法を提供する。これにより、RNA、DNA、RNA−DNAキメラ及び類似体を含むオリゴヌクレオチドを効率的に製造することができる。
以下、本発明を実施例を挙げて説明する。但し、下記実施例は本発明を例示するものに過ぎず、本発明の内容を限定するものではない。
出発物質ポリエチレングリコール2000(Alfa Aesar GmbH & Co.KG、ドイツ)30g(15mmol)をピリジン(pyridine、Sigma Aldrich、米国)270mlに溶かし、これに、トリエチルアミン(Sigma Aldrich、米国)3.55ml(25.5mmol)と4,4’−ジメトキシトリチルクロライド(GL biochem、中国)7.12g(21mmol)を入れて常温で20時間反応させた。反応が終了された反応混合物を濃縮し、エチルアセテート450mlと水450mlで抽出して、減圧濃縮した後真空乾燥して2−[ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)]23g(66%)を得た。
1H NMR(CDCl3);δ1.93(br,1,OH),3.20−3.80(m,186,PEG,DMT−OCH3),6.80−6.83(m,4,DMT),7.19−7.47(m,9,DMT)
前記実施例1−1−1で得られた2−[ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)3.9g(1.672mmol)をピリジン20mlに溶かした後、0℃に冷却した。前記反応物に無水コハク酸(succinic anhydride、Acros Organics、米国)351mg(3.512mmol)とDMAP(4−dimethylaminopyridine、Sigma Aldrich、米国)42.5mg(0.334mmol)を添加し、50℃で3日間撹拌した後反応を終了した。反応が終了された反応混合物を減圧濃縮して、コハク酸2−[ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)[化合物A]3.65g(90%、白色固体)を得た。
1H NMR(CDCl3);δ2.65(m,2,CH2CO),3.20−3.88(m,186,PEG,DMT−OCH3),4.25(m,2,CH2CO),6.80−6.82(m,4,DMT),7.19−7.47(m,9,DMT)。
前記実施例1−1−2で得られた化合物A1g(0.411mmol)をメチレンクロライド(methylene chloride、大然化学、韓国)20mlに溶かした後、0℃に冷却した。前記反応物にトリエチルアミン143μl(1.03mmol)を入れて、クロロギ酸4−ニトロフェニル(4−nitro phenyl chloroformate)149mg(0.740mmol)を添加し、30分間常温に温度を上げて、4時間撹拌した後反応を終了した。反応が終了された反応混合物をNaHCO3飽和水溶液20mlと0℃〜4℃に冷却した1Mクエン酸(Sigma Aldrich、米国)5mlで1回洗浄した後、Na2SO4(Samchum Chemical Co.,韓国)で乾燥した。これを濾過フラスコ(filtering flask)、吸引濾過器(buchner funnel)、真空装置(aspirator)を利用して濾過した後減圧濃縮し、パラ−ニトロフェニルコハク酸2−ビス−(ジメトキシトリチル)−ポリ(エチレングリコール)(p−Nitrophenylsuccinic acid 2−[bis−(4−dimethoxytrityl)−poly(ethylene glycol)[化合物B]1.0g(94%、淡色固体)を得た。
1H NMR(CDCl3);δ2.80−2.90(m,2,CH2CO),3.20−3.87(m,186,PEG,DMT−OCH3),4.25(m,2,CH2CO),6.80−6.82(m,4,DMT),7.19−7.47(m,9,DMT)
前記実施例1−1−2で得られた化合物A500mg(0.206mmol)をメチレンクロライド10mlに溶かした後、ピリジン83.14μl(1.03mmol)を入れた。これに、N−スクシンイミジルトリフルオロ酢酸(N−succinimidyl trifluoro acetic acid、Sigma Aldrich、米国)165mg(0.781mmol)を添加し、常温で7時間撹拌した後反応を終了した。反応が終了された反応混合物を減圧濃縮し、2,5−ジオキソ−ピロリジン−1−イルエステルコハク酸2−ビス−(4−ジメトキシトリチル)−ポリ(エチレングリコール)(2,5−dioxo−pyrrolidin−1−yl ester succinic acid2−[bis−(4−dimethoxytrityl)−poly(ethyleneglycol)[化合物C]490mg(94%、白色固体)を得た。
1H NMR(CDCl3);δ2.72−2.97(m,6,CH2CO,CH2CH2),3.20−3.87(m,186,PEG,DMT−OCH3),4.27−4.28(m,2,CH2CO),6.80−6.83(m,4,DMT),7.20−7.47(m,9,DMT)
直径40〜75μm、空隙サイズ2000ÅのCPG(Silicycle Inc.、カナダ)10gをトルエン100mlに均一に混合して濡らした後、3−アミノプロピルトリエトキシシラン(3−aminopropyltriethoxysilane、TCI Org.Chem、日本)2mlを入れて混合し、常温で8時間反応させた。反応が終了された混合物を濾過し、メタノール、水、メチレンクロライドで順に洗浄した後、真空乾燥してLCAA−CPG(2000Å)10gを得た。
前記実施例1−2−1で得られたLCAA−CPG(2000Å)2gをメチレンクロライド20mlで濡らしておいた。また、化合物A80mgとTEA(Triethylamine、Sigma Aldrich、米国)14μl、BOP(Benzortiazol−1−yloxytris(dimethylamino)phosphonium hexafluorophosphate、TCI Org.Chem、日本)15mg、HOBT(1−Hydroxybenzotriazole anhydrous、TCI Org.Chem、日本)5mgをメチレンクロライド2mlに溶かした溶液を、前記LCAA−CPG(2000Å)溶液と均一に混合した。これを8時間還流反応させた後、反応が終了された混合物を濾過し、メタノール、水、メチレンクロライドで順に洗浄した後真空乾燥した。
sense 5’−AAGGAGAUCAACAUUUUCA(dTdT)−PEG(6664.96Da+2000Da)(配列番号1)
antisense 5’−UGAAAAUGUUGAUCUCCUU(dTdT)−PEG(6592.84Da+2000Da)(配列番号5)
sense 5’−AAGGAGAUCAACAUUUUCA(dTdT)−PEG(6664.96Da+2000Da)(配列番号1)
antisense 5’−UGAAAAUGUUGAUCUCCUU(dTdT)−PEG(6592.84Da+2000Da)(配列番号5)
以下の実施例では、サバイビンを抑制するためにサバイビンsiRNAを用いた。サバイビンは、今までテストされた殆どの新生腫瘍や形質変異された細胞株で共通に発現されるタンパク質であり、坑癌治療において重要なターゲットとなると予測されている(Abbrosini G.et al.Nat.med.3(8):917−921,1997)。本発明のサバイビンsiRNAの配列は、19個のヌクレオチドで構成された場合、配列番号1番に記載されるセンス鎖とこれに相補的な配列のアンチセンス鎖で構成されている。その他に、23個、27個、31個のヌクレオチドで構成された場合、配列番号2、3、4に記載される塩基配列を有する。
(配列番号2)5’−AGGAAAGGAGAUCAACAUUUUCA−3’
(配列番号3)5’−AGGAAAGGAGAUCAACAUUUUCAAAUU−3’
(配列番号4)5’−AAAGGAGAUCAACAUUUUCAAAUUAGAUGUU−3’
前記実施例2で製造、分離したsiRNA高分子化合物接合体が、高分子化合物が結合されていない元来のsiRNAに比べ安定性が向上されたかを確認した。修飾されていない元来のsiRNAと前記実施例2で製造したsiRNA高分子化合物接合体1〜5を、生体内条件を模倣した形態である10%のFBS(fetal bovine serum)が添加された培地でそれぞれ0、1、3、6、9、12、24、36または48時間培養した後、siRNAの分解程度を電気泳動で検討した。
siRNA高分子化合物接合体9〜14の場合、siRNAの末端に付加された疎水性高分子化合物間の疎水性相互作用により、siRNA高分子化合物接合体からなるナノ粒子、即ち、ミセル(micelle)が形成される(図10)。ゼータ電位測定器(zeta−potential measurement)を利用して前記ナノ粒子のサイズを測定した。
前記実施例2で製造したそれぞれのsiRNA高分子化合物接合体1〜8を利用して、腫瘍細胞株であるヒト子宮癌細胞株をトランスフェクションさせ、前記トランスフェクションされた腫瘍細胞株でサバイビン遺伝子の発現様相を分析した。
アメリカ培養細胞系統保存機関(American type Culture Collection、ATCC)から得られたヒト子宮癌細胞(HeLa)は、RPMI1640培養培地(GIBCO、Invitorgen、米国)に10%(v/v)ウシ胎児血清、ペニシリン100units/ml、ストレプトマイシン100μg/mlを添加して、37℃、5%(v/v)CO2の条件下で培養した。
前記実施例2で製造された配列番号1のsiRNA高分子化合物接合体1〜8を利用して、腫瘍細胞株であるヒト子宮癌細胞株(HeLa)をトランスフェクションさせ、前記トランスフェクションされた腫瘍細胞株で、サバイビンの発現様相を分析した。
前記実施例5−1で培養された腫瘍細胞株1.3×105を、37℃、5%(v/v)CO2の条件で6−ウェルプレートで18時間RPMI1640で培養した後培地を除去し、各ウェル当り800μlのOpti−MEM培地(GIBCO、米国)を分注した。
前記実施例5−2−1でトランスフェクションされた細胞株から全体RNAを抽出してcDNAを製造した後、リアルタイムPCR(real−time PCR)を利用してサバイビン遺伝子のmRNA量を相対定量した。
RNA抽出キット(AccuPrep Cell total RNA extraction kit、BIONEER、韓国)を利用して前記実施例5−2−1でトランスフェクションされた細胞株から全体RNAを抽出し、抽出されたRNAは、RNA逆転写酵素(AccuPower CycleScript RT Premix/dT20、BIONEER、韓国)を利用して次の方法によりcDNAを製造した。
前記実施例5−2−2−1で製造されたcDNAを鋳型とし、リアルタイムPCRを利用してサバイビンmRNAの相対的量を次の方法により定量した。
siRNA配列番号1〜4の塩基配列に、siRNA高分子化合物接合体4の構造に末端修飾を誘導したsiRNAを利用して、トランスフェクション物質とともにsiRNA親水性高分子化合物接合体で細胞をトランスフェクションさせた時の標的遺伝子のmRNA発現の阻害を分析した。
前記実施例5−1で培養された腫瘍細胞株1.3×105を、前記37℃、5%(v/v)CO2の条件下で6−ウェルプレートで24時間RPMI1640で培養した後培地を除去し、各ウェル当り800μlのOpti−MEM培地を分注した。
前記実施例5−3−1でトランスフェクションされた細胞株から全体RNAを抽出してcDNAを製造した後、リアルタイムPCR(Real−time PCR)を利用してサバイビン遺伝子のmRNA量を相対定量した。
RNA抽出キット(AccuPrep Cell total RNA extraction kit、BIONEER、韓国)を利用して前記実施例5−3−1でトランスフェクションされた細胞株から全体RNAを抽出し、抽出されたRNAは、RNA逆転写酵素(AccuPower CycleScript RT Premix/dT20、BIONEER、韓国)を利用して次の方法によりcDNAを製造した。
前記実施例5−3−2−1で製造されたcDNAを鋳型とし、リアルタイムPCRを利用してサバイビンmRNAの相対的な量を次の方法により定量した。
前記実施例2で製造したそれぞれのsiRNA高分子化合物接合体1〜14を利用して、腫瘍細胞株であるヒト子宮癌細胞株(HeLa)をトランスフェクションさせ、前記トランスフェクションされた腫瘍細胞株のサバイビンの発現様相を分析した。
アメリカ培養細胞系統保存機関(American type Culture Collection、ATCC)から得られたヒト子宮癌細胞(HeLa)は、RPMI1640培養培地(GIBCO/Invitorgen、米国)に10%(v/v)ウシ胎児血清、ペニシリン100units/ml、ストレプトマイシン100μg/mlを添加して、37℃、5%(v/v)CO2の条件下で培養した。
前記実施例6−1で培養された腫瘍細胞株1.3×105を、37℃、5%(v/v)CO2の条件で6−ウェルプレートで24時間RPMI1640で培養した後培地を除去し、各ウェル当り900μlのOpti−MEM培地を分注した。
前記実施例6−2でトランスフェクションされた細胞株から全体RNAを抽出してcDNAを製造した後、リアルタイムPCR(Real−time PCR)を利用してサバイビン遺伝子のmRNA量を相対定量した。
RNA抽出キット(AccuPrep Cell total RNA extraction kit、BIONEER、韓国)を利用して前記実施例6−2でトランスフェクションされた細胞株から全体RNAを抽出し、抽出されたRNAは、RNA逆転写酵素(AccuPower CycleScript RT Premix/dT20、BIONEER、韓国)を利用して次の方法によりcDNAを製造した。
前記実施例6−3−1で製造されたcDNAを鋳型とし、リアルタイムPCRを利用してサバイビンmRNAの相対的な量を次の方法により定量した。
Claims (14)
- 下記構造のsiRNA−高分子化合物接合体。
A−X−R−Y−B
(上記で、A及びBの一方が親水性高分子化合物であり、他方が、分子量250〜1,000の、C 16 −C 50 の炭化水素またはコレステロールである疎水性化合物であり、X、Yはそれぞれ独立に、リンカーで媒介されていない共有結合、またはリンカーで媒介された共有結合であり、RはsiRNAを示す。) - 親水性高分子化合物及び疎水性化合物が、siRNAの同じ鎖に接合している請求項1に記載の接合体。
- 親水性高分子化合物及び疎水性化合物が、siRNAの両方の鎖に接合している請求項1に記載の接合体。
- siRNA(R)の一本鎖は、19〜31個のヌクレオチドで構成される請求項1〜3のいずれかに記載の接合体。
- 共有結合(X、Y)は、非分解性結合または分解性結合である請求項1〜4のいずれかに記載の接合体。
- 非分解性結合は、アミド結合またはリン酸結合である請求項5に記載の接合体。
- 分解性結合は、ジスルフィド結合、酸分解性結合、エステル結合、アンヒドリド結合、生分解性結合及び酵素分解性結合から選択される請求項5に記載の接合体。
- 親水性高分子化合物は、分子量1,000〜10,000の非イオン性高分子化合物である請求項1〜7のいずれかに記載の接合体。
- 親水性高分子化合物は、ポリエチレングリコール(PEG)、ポリビニルピロリドン、ポリオキサゾリンからなる群から選択される請求項1〜8のいずれかに記載の接合体。
- 請求項1〜9のいずれかに記載の接合体の製造方法であって、
1)以下の構造を有するポリエチレングリコール(PEG)が結合された固形支持体を利用して、siRNA−PEG接合体の一本鎖を製造する段階と、
(上記で、Rは、アルキル、アルケニル、アルキニル、アリール、アリールアルキル、ヘテロアルキル、またはヘテロアリールであり;mは、2〜18の整数であり、nは5〜120の整数であり、Xは、水素、4−モノメトキシトリチル基、4,4’−ジメトキシトリチル基、または4,4’,4”−トリメトキシトリチル基である。)
2)分子量250〜1,000の、C 16 −C 50 の炭化水素またはコレステロールである疎水性化合物を、前記PEGが接合した鎖に遠位末端において共有結合で連結する段階と、
3)両端部でPEG及び前記疎水性化合物が接合している前記鎖を、その相補鎖にアニーリングする段階と
を含む、製造方法。 - 請求項1〜9のいずれかに記載の接合体の製造方法であって、
1)siRNAの一方の鎖の末端基をポリエチレングリコール(PEG)又は分子量250〜1,000の、C 16 −C 50 の炭化水素またはコレステロールである疎水性化合物に共有結合で連結する段階と、
2)前記siRNAの他方の鎖の末端基を、段階1)で疎水性化合物を連結した場合には、PEGに、又は段階1)でPEGを連結した場合には、疎水性化合物に、共有結合で連結する段階と、
3)前記段階1)で得た鎖を前記段階2)で得た他方の鎖にアニーリングする段階と
を含む、製造方法。 - 請求項1〜9のいずれかに記載の接合体で構成されたナノ粒子。
- 請求項1〜9のいずれかに記載の接合体の薬学的有効量を含む薬学的組成物。
- 請求項12に記載のナノ粒子の薬学的有効量を含む薬学的組成物。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2015508998A (ja) * | 2012-01-05 | 2015-03-26 | バイオニア コーポレーションBioneer Corporation | 高効率のナノ粒子型二本鎖オリゴrna構造体およびその製造方法 |
Families Citing this family (36)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101224828B1 (ko) | 2009-05-14 | 2013-01-22 | (주)바이오니아 | siRNA 접합체 및 그 제조방법 |
BR112014014730B1 (pt) * | 2011-12-15 | 2021-05-18 | Bioneer Corporation | estrutura de droga polímero terapêutica, métodos para preparar uma estrutura de oligo rna de dupla hélice, nanoparticula e composição farmacêutica |
KR101722948B1 (ko) * | 2012-01-05 | 2017-04-04 | (주)바이오니아 | 리간드가 결합된 이중나선 올리고 rna 구조체 및 그 제조방법 |
EP2805713B1 (en) * | 2012-01-18 | 2018-10-10 | Bioneer Corporation | Magnetic nanoparticle-samirna complex and method for preparing same |
RU2015114330A (ru) | 2012-09-17 | 2016-11-10 | У.Р. Грейс Энд Ко.-Конн. | Хроматографические среды и устройства |
US20150259690A1 (en) * | 2012-10-05 | 2015-09-17 | Bioneer Corporation | Amphiregulin-specific double-helical oligo-rna, double-helical oligo-rna structure comprising double-helical oligo-rna, and composition for preventing or treating respiratory diseases containing same |
WO2014088087A1 (ja) * | 2012-12-06 | 2014-06-12 | 協和発酵バイオ株式会社 | アジュバント用二重鎖リボ核酸 |
PL3018208T3 (pl) * | 2013-07-05 | 2020-09-21 | Bioneer Corporation | Ulepszona struktura oligonukleotydowa typu nanocząstki o wysokiej wydajności i sposób jej wytwarzania |
CN110592082A (zh) * | 2013-07-05 | 2019-12-20 | 柏业公司 | 呼吸疾病相关基因特异性siRNA、含有siRNA的双螺旋寡RNA结构及其用途 |
SG11201507571TA (en) * | 2013-07-05 | 2015-10-29 | Bioneer Corp | Dengue virus-specific sirna, double helix oligo-rna structure comprising sirna, and composition for suppressing proliferation of dengue virus comprising rna structure |
KR20150006742A (ko) * | 2013-07-09 | 2015-01-19 | (주)바이오니아 | 간암 연관 유전자 특이적 siRNA, 그러한 siRNA를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 암 예방 또는 치료용 조성물 |
KR20150006743A (ko) * | 2013-07-09 | 2015-01-19 | (주)바이오니아 | 간암 연관 유전자 특이적 siRNA, 그러한 siRNA를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 암 예방 또는 치료용 조성물 |
CN106459974A (zh) * | 2014-04-04 | 2017-02-22 | 柏业公司 | 新颖的双链寡rna和包含它的用于预防或治疗纤维化或呼吸系统疾病的药物组合物 |
EP3137209B1 (en) | 2014-05-02 | 2022-10-05 | W.R. Grace & CO. - CONN. | Functionalized support material and methods of making and using functionalized support material |
EP3207069B1 (en) | 2014-10-15 | 2021-03-10 | University Of Connecticut | Bio-reducible self-assembled liquid crystalline block copolymer for drug delivery |
PL3302784T3 (pl) | 2015-06-05 | 2022-01-17 | W.R. Grace & Co.-Conn. | Adsorbentowe środki klarujące do bioprzetwarzania oraz sposoby ich wytwarzania i stosowania |
WO2016204515A1 (ko) * | 2015-06-15 | 2016-12-22 | (주)바이오니아 | STAT3 유전자 특이적 siRNA, 그러한 siRNA를 포함하는 이중나선 올리고 RNA 구조체, 이를 포함하는 조성물 및 이의 용도 |
KR101861738B1 (ko) | 2016-08-24 | 2018-05-29 | (주)바이오니아 | 마이크로 rna를 포함하는 이중나선 올리고 rna 구조체 |
EP3600439A4 (en) * | 2017-03-22 | 2021-01-13 | The Regents of the University of California | MODIFIED OLIGONUCLEOTIDES AND THEIR THERAPEUTIC USES |
US11414665B2 (en) | 2017-12-01 | 2022-08-16 | Suzhou Ribo Life Science Co., Ltd. | Nucleic acid, composition and conjugate comprising the same, and preparation method and use thereof |
CN118291456A (zh) | 2017-12-01 | 2024-07-05 | 苏州瑞博生物技术股份有限公司 | 一种核酸、含有该核酸的组合物与缀合物及制备方法和用途 |
WO2019105418A1 (zh) | 2017-12-01 | 2019-06-06 | 苏州瑞博生物技术有限公司 | 双链寡核苷酸、含双链寡核苷酸的组合物与缀合物及制备方法和用途 |
JP7360716B2 (ja) * | 2017-12-01 | 2023-10-13 | スーチョウ リボ ライフ サイエンス カンパニー、リミテッド | 核酸、当該核酸を含む組成物および複合体ならびに調製方法と使用 |
AU2018394875B2 (en) | 2017-12-29 | 2023-08-03 | Suzhou Ribo Life Science Co., Ltd. | Conjugates and preparation and use thereof |
KR102141124B1 (ko) | 2018-01-30 | 2020-08-04 | (주)바이오니아 | 이중 가닥 miRNA를 포함하는 이중나선 올리고뉴클레오타이드 구조체 및 이의 용도 |
US20210189398A1 (en) | 2018-05-25 | 2021-06-24 | Bioneer Corporation | Amphiregulin gene-specific double-stranded oligonucleotide and composition for preventing and treating fibrosis-related diseases and respiratory diseases, comprising same |
EP3842534A4 (en) | 2018-08-21 | 2022-07-06 | Suzhou Ribo Life Science Co., Ltd. | NUCLEIC ACID, COMPOSITION AND CONJUGATE CONTAINING NUCLEIC ACID AND METHOD OF USE THEREOF |
CN111655297A (zh) * | 2018-09-30 | 2020-09-11 | 苏州瑞博生物技术有限公司 | 一种siRNA缀合物及其制备方法和用途 |
KR102473989B1 (ko) | 2018-11-28 | 2022-12-07 | (주)바이오니아 | 안드로젠 수용체 특이적 서열을 포함하는 이중나선 올리고뉴클레오티드 구조체, 및 이를 포함하는 탈모 예방 및 발모용 조성물 |
WO2020149644A1 (ko) | 2019-01-15 | 2020-07-23 | (주)바이오니아 | Dkk1 유전자를 표적으로 하는 이중나선 올리고뉴클레오티드, 이를 포함하는 구조체 및 이를 포함하는 탈모 예방 또는 발모용 조성물 |
KR20210063137A (ko) | 2019-11-22 | 2021-06-01 | (주)바이오니아 | Ctgf 유전자 특이적 이중가닥 올리고뉴클레오티드 및 이를 포함하는 섬유증 관련 질환 및 호흡기 관련 질환 예방 및 치료용 조성물 |
US20230348912A1 (en) | 2020-05-14 | 2023-11-02 | Bioneer Corporation | Composition for preventing or treating obesity-related disease containing amphiregulin-specific double-stranded oligonucleotide structure |
JP2023533124A (ja) | 2020-05-22 | 2023-08-02 | バイオニア コーポレーション | 二本鎖オリゴヌクレオチド及びこれを含むコロナウイルス感染症-19(covid-19)治療用組成物 |
KR102272800B1 (ko) | 2020-06-30 | 2021-07-05 | 국방과학연구소 | 코로나바이러스 특이적 이중가닥 올리고뉴클레오티드 및 이를 포함하는 코로나바이러스 감염증-19 예방 및 치료용 조성물 |
CA3206861A1 (en) | 2021-02-25 | 2022-09-01 | Han-Oh Park | Composition for alleviating hair graying, promoting hair growth and/or preventing or alleviating hair loss, comprising double-stranded mirna as active ingredient |
WO2022191567A1 (ko) | 2021-03-08 | 2022-09-15 | (주)바이오니아 | Covid-19를 포함하는 호흡기 바이러스 감염증, 바이러스 감염에 의한 폐섬유증, 또는 호흡기 질환 예방 또는 치료를 위한 초음파 방식 연무식 흡입기를 이용한 이중가닥 올리고뉴클레오티드 구조체 투여용 조성물 |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5451463A (en) * | 1989-08-28 | 1995-09-19 | Clontech Laboratories, Inc. | Non-nucleoside 1,3-diol reagents for labeling synthetic oligonucleotides |
US5141813A (en) * | 1989-08-28 | 1992-08-25 | Clontech Laboratories, Inc. | Multifunctional controlled pore glass reagent for solid phase oligonucleotide synthesis |
KR100270195B1 (ko) * | 1991-09-06 | 2001-03-02 | 와일러 제임스 에프. | 겔로닌 폴리펩타이드를 암호화하는 dna 서열 |
US6221959B1 (en) * | 1994-11-18 | 2001-04-24 | Supratek Pharma, Inc. | Polynucleotide compositions |
US6348583B1 (en) * | 1999-08-30 | 2002-02-19 | Bio-Rad Laboratories, Inc. | Poly(ether-thioether), poly(ether-sulfoxide) and poly(ether-sulfone) nucleic acids |
US7491805B2 (en) * | 2001-05-18 | 2009-02-17 | Sirna Therapeutics, Inc. | Conjugates and compositions for cellular delivery |
DK2284266T3 (da) | 2002-11-14 | 2014-01-13 | Thermo Fisher Scient Biosciences Inc | sIRNA-MOLEKYLE MOD TP53 |
AU2003219576A1 (en) * | 2003-04-03 | 2004-10-25 | Korea Advanced Institute Of Science And Technology | Conjugate for gene transfer comprising oligonucleotide and hydrophilic polymer, polyelectrolyte complex micelles formed from the conjugate, and methods for preparation thereof |
US7851615B2 (en) * | 2003-04-17 | 2010-12-14 | Alnylam Pharmaceuticals, Inc. | Lipophilic conjugated iRNA agents |
CN1984921B (zh) * | 2003-06-03 | 2010-06-16 | Isis药物公司 | 存活蛋白表达的调节 |
KR101147147B1 (ko) * | 2004-04-01 | 2012-05-25 | 머크 샤프 앤드 돔 코포레이션 | Rna 간섭의 오프 타겟 효과 감소를 위한 변형된폴리뉴클레오타이드 |
CA2619533C (en) | 2005-08-17 | 2014-02-04 | Bioneer Corporation | Sirna-hydrophilic polymer conjugates for intracellular delivery of sirna and method thereof |
WO2007051303A1 (en) | 2005-11-02 | 2007-05-10 | Protiva Biotherapeutics, Inc. | MODIFIED siRNA MOLECULES AND USES THEREOF |
EP2046954A2 (en) * | 2006-07-31 | 2009-04-15 | Curevac GmbH | NUCLEIC ACID OF FORMULA (I): GIXmGn, OR (II): CIXmCn, IN PARTICULAR AS AN IMMUNE-STIMULATING AGENT/ADJUVANT |
EP2025348A1 (en) * | 2007-08-13 | 2009-02-18 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Targeted block copolymer micelles |
WO2009039173A2 (en) * | 2007-09-19 | 2009-03-26 | Applied Biosystems Inc. | SiRNA SEQUENCE-INDEPENDENT MODIFICATION FORMATS FOR REDUCING OFF-TARGET PHENOTYPIC EFFECTS IN RNAi, AND STABILIZED FORMS THEREOF |
TW200927177A (en) * | 2007-10-24 | 2009-07-01 | Nat Inst Of Advanced Ind Scien | Lipid-modified double-stranded RNA having potent RNA interference effect |
KR101224828B1 (ko) | 2009-05-14 | 2013-01-22 | (주)바이오니아 | siRNA 접합체 및 그 제조방법 |
-
2009
- 2009-05-14 KR KR1020090042297A patent/KR101224828B1/ko active IP Right Grant
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Publication number | Priority date | Publication date | Assignee | Title |
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JP2015508998A (ja) * | 2012-01-05 | 2015-03-26 | バイオニア コーポレーションBioneer Corporation | 高効率のナノ粒子型二本鎖オリゴrna構造体およびその製造方法 |
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