JP5980467B2 - Il−7薬物原料、組成物、製造及び使用 - Google Patents
Il−7薬物原料、組成物、製造及び使用 Download PDFInfo
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Description
B及びTリンパ球は免疫応答の一次エフェクター細胞である。両細胞クラスは、各クラスの分化における識別可能な工程を表す前駆細胞(progenitor又はprecursor cells)を経由して哺乳動物骨髄における造血幹細胞から最終的に誘導されるものと考えられる。成熟T細胞は、多分、Tリンパ球発生の初期工程で骨髄から胸腺に移行する前駆細胞から、主として胸腺で発生する。IL−1、IL−2、IL−4、IL−5、インターフェロンγ、BSF−2、ニューロロイキン(neuroleukin)、トランスフォーミング増殖因子β及びIL−7を含む多数の因子が成熟した末梢B及びT細胞に対して活性である(EP0314415)。
本発明の第1の目的は、下記の3つのジスルフィド架橋:Cys:1−4(Cys2−Cys92);2−5(Cys34−Cys129);3−6(Cys47−Cys141)を含む精製されたIL−7コンホーマーに関する。更に下記に開示されたとおり、IL−7コンホーマーは好ましくはヒト組換えIL−7コンホーマーであり、そしてグリコシル化されていてもよく又はグリコシル化されていなくてもよい。
a)IL−7ポリペプチドを含むサンプルを提供し、そして
b)上記したIL−7コンホーマーを精製する、
ことを含む。
本発明は、下記の3つのジスルフィド架橋:Cys:1−4(Cys2−Cys92);2−5(Cys34−Cys129);3−6(Cys47−Cys141)を含む精製されたIL−7コンホーマー、該IL−7コンホーマーを含む薬物原料及び製薬学的組成物、並びにそれらの製造方法及び治療的使用に関する。更に下記に開示されるとおり、IL−7コンホーマーは好ましくはヒト組換えIL−7コンホーマーでありそしてグリコシル化されていても良くそしてグリコシル化されていなくてもよい。ヒト組換えIL−7コンホーマーはグリコシル化されているのが好ましい。
本発明の第一の目的は、下記の3つのジスルフィド架橋:Cys:1−4(Cys2−Cys92);2−5(Cys34−Cys129);3−6(Cys47−Cys141)を含む精製された又は単離されたIL−7コンホーマーにある。
が可能であり、その中でも1−6;2−5;3−4又は1−4;2−5;3−6又は1−4;2−6;3−5又は1−5;2−4;3−6、に加えて、連続的架橋(contiguous bridges)を含む種々の他のコンホーマーが可能である。
本発明は、所望される生成物として上記したIL−7コンホーマーを含む薬物原料であって、IL−7生成物関連物質及び生成物関連不純物を実質的に含まない薬物原料にも関する。
本発明の他の観点は、上記組成物、特に上記IL−7コンホーマー及び薬物原料をその製薬学的使用のために十分な量及び品質において製造するための適当な構築物及び方法を提供することである。
i)IL−7遺伝子又はコード配列を含有するサンプルを提供し、
ii)該サンプルを、最適化された配列、特に位置49における機能的第2開始コドンを欠いた配列を生成するプライマーの対と接触させ、そして
iii)最適化されたIL−7核酸を生成又は増幅する、
ことを含む方法も提供する。
対1:配列番号5及び配列番号6、
対2:配列番号7及び配列番号6、及び
対3:配列番号8及び配列番号6。
a)IL−7ポリペプチドを含むサンプルを提供し、そして
b)上記したIL−7コンホーマーを精製する、
ことを含む。
i)該サンプルを処理して該IL−7ポリペプチドの完全な変性を引起し、
ii)場合により工程i)で得られた変性されたポリペプチドを精製し、そして
iii)ポリペプチドをリフォールディングする、
ことを含む。
は好ましくはデキストラン硫酸又はヘパリンであり、そしてマトリックスはセファロース、アクリルアミド、アガロース、デキストラン、セルロース又はタンパク質精製でよく使用される他のタイプのものである。好ましいマトリックスはヘパリンセファロースである。この方法は、更に所望のIL−7コンホーマーを溶離する追加の工程を含むことができる。
精製工程b)は、医薬関連物質、例えば、他のコンホーマー、脱アミド化形態、タンパク質の二量体、DNA、内毒素等を含む残留不純物を除去するために、1つ又はそれより多くの精製工程、例えば、ろ過又は限外ろ過工程、イオン交換クロマトグラフィー、アフィニティークロマトグラフィー、疎水性相互作用クロマトグラフィー等を含むことができる。クロマトグラフィー工程は、フロースルー方式(flow through mode)、捕集方式(capture mode)又は流線形方式(streamline mode)で達成することができる。
a)組換え原核生物宿主細胞により産生されたIL−7ポリペプチドを含むサンプルを提供し、
a′)該サンプルを処理して該IL−7ポリペプチドの完全な変性を引起し、
a″)場合により工程a″)で得られた変性されたポリペプチドを精製し、
a″′)ポリペプチドをリフォールディングし、そして
b)ポリペプチドを精製して上記したコンホーマーを生成させる、
工程を含む方法に関する。
a)組換え真核生物宿主細胞により産生されたサンプルであって、リフォールディングされたIL−7ポリペプチドを含むサンプルを提供し、そして
b)生成物関連物質又は不純物を除去するために1つ又はそれより多くの精製工程を実施する、
工程を含む方法に関する。
実施例A.最適化されたヒト(h)及びサル(s)IL−7コードヌクレオチド配列の構築及び発現
A1.大腸菌発現(E.coli JM101)
1.1.ヒトIL−7コードヌクレオチド配列の構築
ヒトIL−7コードcDNA配列を、制限エンドヌクレアーゼ認識配列を含有する下記の特異的オリゴヌクレオチドプライマー:
−配列番号:5:IL75′
5′ATTCCATATGGATTGTGATATTGAAGGTAAAGATGGC3′
NdeI
−配列番号:6:IL73′
5′AGCCGGATCCTTATCAGTGTTCTTTAGTGCCCATCA3′
BamHI
を使用してヒト胎盤cDNA(BioChain Inc)からポリメラーゼ連鎖反応(PCR)(Mullis et al.; 1987; Methods in Enzymology; 155: 335-350)により増幅した。
−配列番号:7:mutlL75′
5′TAGGGAATTCCATATGGATTGTGATATTGAAGGTAAAG
Ndel
ATGGCAAACAATACGAGTCCGTTCTG3′
−配列番号:6:IL73′
5′AGCCGGATCCTTATCAGTGTTCTTTAGTGCCCATCA3′
BamHI
を使用して部位特異的突然変異誘発PCRにより行った。
配列番号:9:ptac1
5′ATCGAGATCTAATTCTCATGTTTGACAGCTTATCAT3′
BgIII
配列番号:10:ptac2
5′ATCGTCTAGAGCTGTTTCCTGTGTGAAATTGTTATCCG
XbaI
3′
を使用してtacプモーターを最初に増幅した。
配列番号:11:ptacプロモータープライマー
5′TTCGTGTCGCTCAAGGCGCA3′
を使用する配列決定分析により正しいDNA配列が確認された。
1.2 サルIL−7コードヌクレオチド配列の構築
a)5′領域を含むサルIL−7cDNAの増幅及び配列決定
サル(アカゲザル(Macaca Mulatta Rheusus monkey))IL−7コードcDNA配列は、PCR増幅(Mullis et al.; 1987; Methods in Enzymology; 155: 335-350)を使用してアカゲザル腎臓cDNA(BioChain Inc)から得られた。基本的ストラテジーは、ヒトIL−7cDNAを増幅するのに使用した特定のオリゴヌクレオチドプライマー(配列番号5:IL75′及び配列番号6:IL73′)によるPCRによりサルIL−7cDNAを増幅することであった。ゲル電気泳動で1つのバンドが現れた(図6参照)。非対称PCRs及び配列決定はサルIL−7配列を得ることを可能とする。ヒト及びサルIL−7DNA及びアミノ酸配列間の配列相同分析は、それぞれ、DNA(図4参照)配列で98.1%の相同性−同一性、及びアミノ酸(図5参照)配列で96.6%の相同性−同一性を示した。
ヒトIL−7配列と同様に、サルIL−7コードDNA配列は、「ATG」開始コドンの後の位置49に、大腸菌における第2開始コドンとして挙動することができる第2推定「ATG」を与える。何故ならば、この第2「ATG」は、「擬シャイン・ダルガノ」配列(リボソーム結合配列)により先行されるからである。r−sIL−7のアミノ末端トランケーションされた形態(amino-terminal truncated form)の生成の可能性を回避するために、「SD様」配列のヌクレオチドのいくらかを然変異させ(得られるコードされたr−sIL−7アミノ酸配列を修飾することなく)、それにより大腸菌細胞中での発現のための1つ又はそれより多くの好ましいコドン(1つ又は複数)が含有する改良されたメチオニル−IL−7コードDNA配列を生成させた。
対象:天然IL−7ペプチドシグナルに5′端部で連結されたIL−7cDNA断片のcDNAの一部を増幅する試みにおけるプライマーとしてオリゴヌクレオチドを使用して、ヒトIL−7コードcDNA配列を、ヒト胎盤cDNA(BioChain Inc)からポリメラーゼ連鎖反応(PCR)(Mullis et al.; 1987; Methods in Enzymology; 155: 335-350)により増幅した。
−配列番号8:PSIL75′
5′GCAAGCTTGCCACCATGTTCCATGTTTCTTTTAGGTAT
HindIII Kozak配列
ATCTTTGGAC3′
−配列番号6:IL−73′
5′AGCCGGATCCTTATCAGTGTTCTTTAGTGCCCATCA3′
BamHI
ヒトIL−7コードcDNA配列を制限エンドヌクレアーゼHindIII及びBamHIによる消化によりpcDNA−hPSIL−7から単離した。得られる「hPSIL−7 cDNA」断片をアガロースゲルで精製し、そして同じ酵素で加水分解されたpcDNA4/TO(Invitrogen)に挿入した。ライゲーション生成物をTOP10F′コンピテント細胞(Invitrogen)中に形質転換した。プラスミド含有細胞の選択は、ベクターに保有された抗生物質(アンピシリン)耐性マーカー遺伝子に基づいていた。ポジティブクローンからのプラスミドDNAを培養された細胞から単離し、制限マッピングによりチェックし、そしてユニバーサルCMVフォーワードプライマー及びBGHリバースプライマーを使用する配列決定分析により確認した。
EcoRVとMluI制限部位間の問題のcDNA(その天然のペプチドシグナルに連結されたIL−7cDNA)を増幅する試みにおいてプライマーとしてオリゴヌクレオチドを使用して、ヒトIL−7コードcDNA配列を、pcDNA−hPSIL−7からポリメラーゼ連鎖反応(PCR)(Mullis et al.; 1987; Methods in Enzymology; 155: 335-350)により増幅した。
−配列番号14:PSIL7EcoRV5′
5′AGATATCAATGTTCCATGTTTCTTTTAGGTA3′
EcoRV
−配列番号15:PSIL7MluI3′
−配列番号16:pCMV5
5′CACTTGAGTGACAATGAC3′
−配列番号17:pSV40polyA
5′TCACTGCATTCTAGTTGT3′
ヒトIL−7コードcDNA配列を制限エンドヌクレアーゼHindIII及びBamHIによる消化によりpcDNA−hPSIL−7から単離した。得られる「hPSIL−7 cDNA」断片をアガロースゲルで精製し、そしてHindIII及びBamHI 制限部位で消化されたpBudCE4.1ベクター(Invitrogen)に、pCMVプロモーターの下流で挿入した。
−配列番号14:BclXL5′NotI
5′TAGCGGCCGCATGTCTCAGAGCAACCGG3′
NotI
−配列番号15:BclXL3′BstBI
5′ACTTCGAATCATTTCCGACTGAAGAGTG3′
BstBI
組換え(ヒト又はサル)IL−7の産生のための醗酵を、実施例A1に記載の発現プラスミドptac−hIL−7又はptac−slL7optで形質転換された大腸菌JM101宿主株を使用して80リットルの醗酵槽(New Brunswick)で行った。
最善の安定なポジティブクローンを、実施例A2における如く、いくつかの培地スクリーニングにより血清なしの懸濁培養に適合させて、高い細胞密度培養における生産性及び増殖のために最適化されたクローンを産生させた。灌流システムを備えた3リットルのバイオリアクター中で細胞培養を行った。細胞を10百万/mlの濃度に増殖させた。反応器を約3L/日の連続灌流速度で10日間操作した。組換えタンパク質を含有するおおよそ30Lの血清なしの培養培地を発生させそしてr−hIL−7の精製のための出発物質として使用した。
収穫された細胞を、実施例Bにおける如く、トリス20mM/EDTA10mM緩衝液(pH8)中に懸濁させそして4℃で45分間16900gで遠心した。2回引き続く洗浄/遠心サイクルの後、封入体画分を回収した。
実施例Cで報告された発現系HEK−293−pcDNA−hPSIL−7の増殖により発生した粗細胞培養流体を、従来の方法又はストリームラインイオン交換法を使用して処理した。
ジエチルアミノエチル(DEAE)の拡大した床に移し、次いでIEX及びHICの組み合わせに移す。仕上げ工程はろ過及び濃縮を含むことができる。従来の方法では、ろ過と濃縮[マイクロフィルトレーション(0.45μm)限外ろ過/ダイアフィルトレーション]工程の組み合わせを使用して粗細胞培養流体を清澄化して生成物を単離した。得られるタンパク質溶液を、種々の組み合わせにおけるイオン交換組み合わせ及びヘパリン.セファロース[ファスト.フロー(Pharmacia)カラム]にローディングして生成物を精製した。
1つの電荷を有するタンパク質の平均質量(M+H)+。測定は17518.4Daの理論値に対して17517.6Daの分子量を与えた。
大腸菌中で発現され(実施例A1)、実施例Dにおける如く精製されそして特徴付けされた哺乳動物(ヒト及びサル)IL−7をアッセイしそしてCBA/C57BLマウスからの骨髄細胞由来の細胞系、プレB細胞系PB1(DSMZ, Deutsche Sammulung von Mikroorganismen und Zellkulturen, Brawnschweig, Germany)と名づけられたIL−7依存性細胞系の増殖を刺激するそれらの能力についてマウスIL−7(R&D System)と比較した。
ヒト組換えIL−7は、サルのモデルに対して異種でありそして組換えヒトIL−7コンホーマー(上記した如き)に対する中和抗体を誘発することができる。これらの抗体は、in vivoで抑制剤として機能することができそして薬物原料の免疫抑制治療効果に寄与することができる。これに基づいて、純粋な(本発明に記載の)r−hIL−7又はr−sIL−7又は次の実施例に記載された「不純物を含んだ」("impure")r−sIL−7で処理された動物の血清及び血漿において抗IL−7抗体を調べた。
ELISAストリッププレートを0.75μg/mlのツイーンを含有するブロッキング緩衝液中に希釈されたr−IL−7(ヒト又はサル)でコーティングした。4℃で一夜インキュベーションした後、プレートの飽和を室温で1時間ブロッキング緩衝液で行った。すべての溶液を除去しそしてプレートを洗浄緩衝液で洗浄した後、プレートをアッセイに使用する用意をした。
次いで、吸光度を492nmで読み取った。
組換え(ヒト又はサル)IL−7をニトロセルロース又はPVDF膜に移した。次いで膜を処理された動物の血清(1/100又は1/200の希釈率)インキュベーションし、又はポジティブコントロールについては抗IL−7抗体(抗ヒトIL−7抗体:AB207 NA、R&R System)とインキュベーションした。溶液を除去した後、膜を洗浄しそして二次抗体:抗サルIgGアルカリホスファターゼコンジュゲート(SIGMA A1929)と、又はポジティブコントロールについては抗ヤギIgGアルカリホスファターゼコンジュゲート(SIGMA A4187)とインキュベーションした。次いで、BCIP/NBT(SIGMA)を使用して顕色(revelation)を行った。
大腸菌中で発現され(実施例A1)、実施例Dにおける如く精製されそして特徴付けされた組換えIL−7(ヒト及びサル)及び「不純物を含んだ」r−sIL−7(実施例Gで定義された)を正常な霊長類におけるin vivo生物学的活性について試験した。この霊長類モデルは、薬物原料のa)長期活性及び分子変異体及び/又は医薬生成物関連不純物で汚染された同じ薬物原料の免疫原性を試験するための唯一の可能なモデルであった。げっ歯類で同じ効果を試験しても結論は出ないであろう。何故ならば、第一に、げっ歯類配列は霊長類配列に比べて重要な欠失を示し、第二に、げっ歯類においては、IL−7はB細胞に対して強いリンパ球生成効果を有し、これはT細胞リンパ球生成効果をマスクしやすい又はこの効果の明らかに特徴的な分析を妨害しやすいからである。この理由で、ヒトにおける免疫原性製剤の非倫理的な試験を回避しながら、霊長類における効果を証明するために、我々はサルIL−7をクローニングし、発現しそして精製しそしてそれをサルにおいて試験しなければならなかった。
● グループ1のサルはプラシボ(IL−7希釈剤)を受け取った。
● グループ2のサルは、皮下注射により一日1回投与される150μg/kgの用量で4週間r−hIL−7(本発明に記載の)を受け取った。
● グループ3のサルは、皮下注射により一日1回投与される150μg/kgの用量で4週間r−sIL−7(本発明に記載の)を受け取った。
● グループ4のサルは、150μg/kgの用量レベルで「不純物を含んだ」s−rIL−7の1日1回の皮下投与を4週間受け取った。
すべての動物を6週間:4週間の処理期間及び2週間の可逆性期間にわたり調べた。
大腸菌中で発現され(実施例A1)、実施例Dにおける如く精製されそして特徴付けされた組換えサルIL−7及び「不純物を含んだ」r−sIL−7(実施例Gで定義された)を免疫抑制された霊長類における中又は長期間in vivo生物学的活性について試験した。若いアカゲザルを3つのグループ(1〜3)において調べた。各グループは3匹の動物を含んでいた。
サルは下記の治療方式を受けた:
●グループ1はプラシボ(IL−7希釈剤)を受け取った。
●グループ2は、皮下注射により一日1回投与される70μg/kgの用量で、r−sIL−7(本発明に記載の)を、照射後14日目に開始して4週間受け取った。
● グループ3は、70μg/kgの用量レベルで「不純物を含んだ」r−sIL−7の1日1回の皮下投与を、照射後14日目に開始して4週間受け取った。
大腸菌中で発現され(実施例A1)、実施例Dにおける如く精製されそして特徴付けされた組換えヒトIL−7(r−hIL−7)を、薬力学的パラメーター及び正常なシノモルグスサルにおける総リンパ球数値に対するin vivo生物学的活性について試験した。
−末梢血中のリンパ球数値、CD3+T細胞、CD4+及びCD8+サブセットの明らかな減少が6時間という早期にそして72時間まで観察された。細胞数値は注射の約96時間後ベースラインの値に戻った。末梢リンパ球数値のこの低下はTリンパ球の血液からリンパ組織に向けての初期IL−7誘発輸送(early IL-7 induced trafficking)と合致している。
−末梢リンパ球によるIL−7受容体α鎖(CD127)発現の著しいしかし一過性の減少が注射の後48時間の期間観察され、これはIL−7の一回の投与に応答するIL−7受容体のダウンモジュレーションを反映する。CD127発現は48時間に再び現れ始め、注射の96時間後ベースラインの値に向けて戻った。
−細胞増殖のマーカーであるKi67発現の遅延したしかし有意な増加が注射後72/96時間までCD4+及びCD8+細胞の両方で観察された。
−アポトーシス抑制のマーカーであるBcl−2の遅延した増加が観察され、IL−7活性に関与していることが証明された。
Claims (26)
- 組換えヒトIL−7及び1種又はそれより多くの製薬学的に適合性又は許容できる担体、賦形剤又は希釈剤を含む製薬学的組成物であって、
組成物中組換えヒトIL−7の総量の少なくとも98重量%が、下記の3つのジスルフィド架橋:Cys:1−4(Cys2−Cys92);2−5(Cys34−Cys129)及び3−6(Cys47−Cys141)を含む組換えヒトIL−7コンホーマーからなる、製薬学的組成物。 - 該IL−7コンホーマーが、配列番号2又は4のアミノ酸配列を含む、請求項1に記載の組成物。
- 該IL−7コンホーマーが、グリコシル化されていない、請求項1に記載の組成物。
- 該IL−7コンホーマーが、グリコシル化されている、請求項1に記載の組成物。
- 該IL−7コンホーマーが、ヘテロ二量体として肝細胞成長因子に会合している、請求項1〜4のいずれか一項に記載の組成物。
- 該IL−7コンホーマーが、ペプチドヒンジ領域によってIgG重鎖のFc部分に機能的に付着しており、該IgGが、ヒトIgG1又はIgG4である、請求項1〜4のいずれか一項に記載の組成物。
- 該IL−7コンホーマーが、ヒト血清アルブミン(HSA)又はHSAの一部に融合タンパク質として機能的に会合している、請求項1〜4のいずれか一項に記載の組成物。
- 別のIL−7コンホーマーを含まない、請求項1〜7のいずれか一項に記載の組成物。
- 該IL−7コンホーマーの総重量が、組成物中IL−7の総量の少なくとも99.5重量%である、請求項1〜8のいずれか一項に記載の組成物。
- スクロース、トレハロース及びアミノ酸から選ばれる製薬学的に適合性の担体を含む、請求項1〜9のいずれかに記載の組成物。
- 適当な緩衝液中に含有されて等張性溶液を形成する製薬学的に適合性の担体を含む、請求項1〜10のいずれかに記載の組成物。
- 該適当な緩衝液が、5〜7.5のpH範囲を有する、請求項11に記載の組成物。
- 該適当な緩衝液が、クエン酸ナトリウム緩衝液及び酢酸アンモニウム緩衝液から選ばれる有機塩である、請求項12に記載の組成物。
- 該組成物が、凍結乾燥された形態である、請求項1〜13のいずれか一項に記載の組成物。
- 該組成物が、タンパク質及び/又は界面活性剤を含む、請求項1〜14のいずれか一項に記載の組成物。
- 組み合わせて使用するため、別々に使用するため又は順次に使用するための、造血細胞成長因子、サイトカイン、抗原及びアジュバントから選ばれる免疫刺激剤、又はそれらの組み合わせを更に含む、請求項1〜15のいずれか一項に記載の組成物。
- 該造血細胞成長因子が、幹細胞因子(SCF)、G−CSF、GM−CSF、Flt−3リガンド、IL−15及びIL−2から選ばれる、請求項16に記載の組成物。
- サイトカインが、γインターフェロン、IL−2、IL−12、RANTES、B7−1、MIP−2及びMIP−1αから選ばれる、請求項16に記載の組成物。
- 該抗原が、合成又は天然ペプチド、組換えタンパク質、殺されたか、不活性化されたか又は弱毒化された病原生成物、脂質、これらの一部及びこれらの組み合わせから選ばれる、請求項16〜18のいずれか一項に記載の組成物。
- 該抗原が、HIV、水痘帯状疱疹ウイルス、インフルエンザウイルス、エプスタイン−バールウイルス、1型又は2型単純ヘルペスウイルス、ヒトサイトメガロウイルス、デングウイルス、A、B、C又はE型肝炎ウイルス、呼吸器合胞体ウイルス、ヒトパピローマウイルス、結核菌、トキソプラズマ及びクラミジア由来の抗原から選ばれる、請求項19に記載の組成物。
- 該アジュバントが、抗原の免疫原性を促進するか又は増加させそしてTh1型免疫応答を誘発することができる任意の物質、混合物、溶質又は組成物から選ばれる、請求項16〜20のいずれか一項に記載の組成物。
- BもしくはTリンパ球発生及び増殖の予防的もしくは治療的刺激のため、包括的もしくは特異的免疫再構成の増強のため、又は液性もしくは細胞性免疫応答の増強のためにヒト患者に投与するための、請求項1〜21のいずれか一項に記載の製薬学的組成物。
- 免疫不全患者の日和見感染を防止するか又は減少させるための、請求項1〜21のいずれか一項に記載の製薬学的組成物。
- ヒト患者において、リンパ球新生刺激を延長するため及び/又は特異的免疫応答を生じさせるため及び/又は特異的免疫応答のレパートリーを広げるための、請求項1〜21のいずれか一項に記載の製薬学的組成物。
- 免疫不全患者、がん患者、移植片を受けている患者、ウイルス又は寄生虫に感染した患者、高齢患者又は低いCD4数値を有するヒト患者に使用するための、請求項22、23又は24に記載の製薬学的組成物。
- IL−7の有効量が、約3〜300μg/kg/日で含まれる使用のための、請求項1〜25のいずれか一項に記載の組成物。
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Families Citing this family (40)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1324779B1 (en) | 2000-09-29 | 2011-07-20 | Schering Corporation | Pegylated interleukin-10 |
CA2487673C (en) * | 2003-12-02 | 2010-11-02 | F. Hoffmann-La Roche Ag | Improved method for the recombinant production and purification of protein kinases |
BRPI0418286A (pt) | 2003-12-30 | 2007-05-02 | Merck Patent Gmbh | proteìnas de fusão de il-7 |
EP1746161A1 (en) * | 2005-07-20 | 2007-01-24 | Cytheris | Glycosylated IL-7, preparation and uses |
US8216821B2 (en) * | 2005-08-23 | 2012-07-10 | National Research Council Of Canada | Regulation of heterologous recombinant protein expression in methylotrophic and methanotrophic bacteria |
EP2032695B1 (en) * | 2006-05-31 | 2018-08-15 | The Regents of The University of California | Cd127 expression inversely correlates with foxp3 and suppressive function of cd4+ tregs |
CA2664304C (en) | 2006-09-28 | 2017-08-22 | Schering Corporation | Use of pegylated il-10 to treat cancer |
EP2649094B1 (en) | 2010-12-10 | 2016-04-27 | University of Central Florida Research Foundation, Inc. | Methods and compositions comprising il-7 receptor ligands |
WO2013017653A1 (en) | 2011-08-03 | 2013-02-07 | Cytheris | Hcv immunotherapy |
EP2945650B1 (en) * | 2012-04-23 | 2023-07-05 | Bharat Biotech International Limited | Rotavirus vaccine compositions and process for preparing the same |
RU2562169C2 (ru) * | 2012-10-29 | 2015-09-10 | Федеральное государственное унитарное предприятие "Государственный научно-исследовательский институт особо чистых биопрепаратов" Федерального медико-биологического агентства | Штамм культивируемых клеток cho-il7/13 - продуцент интерлейкина-7 человека |
EP2986306A4 (en) | 2013-04-18 | 2016-12-07 | Armo Biosciences Inc | METHOD FOR USE OF INTERLEUKIN-10 FOR THE TREATMENT OF ILLNESSES AND SUFFERING |
US9823255B2 (en) | 2013-06-17 | 2017-11-21 | Armo Biosciences, Inc. | Method for assessing protein identity and stability |
ES2862139T3 (es) | 2013-11-11 | 2021-10-07 | Armo Biosciences Inc | Procedimientos de uso de Interleucina 10 para el tratamiento de enfermedades y trastornos |
EP3119412A1 (en) | 2014-03-21 | 2017-01-25 | Boreal Invest | Terminal nanofiltration of solubilized protein compositions for removal of immunogenic aggregates |
WO2015187295A2 (en) | 2014-06-02 | 2015-12-10 | Armo Biosciences, Inc. | Methods of lowering serum cholesterol |
KR20170084033A (ko) | 2014-10-22 | 2017-07-19 | 아르모 바이오사이언시스 인코포레이티드 | 질환 및 장애를 치료하기 위해 인터루킨-10을 사용하는 방법 |
WO2016126615A1 (en) | 2015-02-03 | 2016-08-11 | Armo Biosciences, Inc. | Methods of using interleukin-10 for treating diseases and disorders |
CN107405384A (zh) * | 2015-03-11 | 2017-11-28 | 尼克塔治疗公司 | Il‑7部分与聚合物的缀合物 |
CN107847583A (zh) | 2015-05-28 | 2018-03-27 | 阿尔莫生物科技股份有限公司 | 用于治疗癌症的聚乙二醇化白细胞介素‑10 |
KR101873201B1 (ko) * | 2015-06-11 | 2018-07-02 | 주식회사 제넥신 | 변형된 인터루킨-7 단백질 및 이의 용도 |
KR102386735B1 (ko) * | 2015-11-06 | 2022-04-14 | 주식회사 제넥신 | 변형된 인터루킨-7 융합 단백질의 제형 |
RU2615447C1 (ru) * | 2015-11-13 | 2017-04-04 | Федеральное государственное унитарное предприятие "Государственный научно-исследовательский институт особо чистых биопрепаратов" Федерального медико-биологического агентства | Синтетическая ДНК, кодирующая интерлейкин-7 человека, содержащий ее экспрессионный вектор (варианты), штамм-продуцент интерлейкина-7 человека и способ получения интерлейкина-7 человека |
US11357827B2 (en) | 2015-12-04 | 2022-06-14 | Genexine, Inc. | Method for preventing or treating influenza virus infection using pharmaceutical composition comprising immunoglobulin Fc-fused interleukin-7 fusion protein |
KR20170066265A (ko) | 2015-12-04 | 2017-06-14 | 주식회사 제넥신 | 면역글로불린 Fc가 융합된 인터루킨-7 융합 단백질을 포함하는 사람 파필로마바이러스 유래 질환의 예방 또는 치료용 약학적 조성물 |
WO2018156649A1 (en) * | 2017-02-22 | 2018-08-30 | Flagship Pioneering, Inc. | Compositions of t cell modulator (tcm) molecules and uses thereof |
CN113365650A (zh) | 2018-11-16 | 2021-09-07 | 新免疫技术有限公司 | 用il-7蛋白和免疫检查点抑制剂的组合治疗肿瘤的方法 |
EP3898677A1 (en) | 2018-12-21 | 2021-10-27 | OSE Immunotherapeutics | Bifunctional anti-pd-1/il-7 molecule |
KR102402276B1 (ko) * | 2019-11-15 | 2022-05-26 | 주식회사 제넥신 | 변형된 인터루킨-7 및 tgf 베타 수용체 ii를 포함하는 융합단백질 및 이의 용도 |
WO2021146191A1 (en) | 2020-01-13 | 2021-07-22 | Neoimmunetech, Inc. | Method of treating a tumor with a combination of il-7 protein and a bispecific antibody |
JP2023512657A (ja) | 2020-02-05 | 2023-03-28 | ワシントン・ユニバーシティ | Il-7タンパク質とcar保有免疫細胞の組み合わせで固形腫瘍を治療する方法 |
US20230398184A1 (en) | 2020-10-26 | 2023-12-14 | Neoimmunetech, Inc. | Methods of inducing stem cell mobilization |
KR20230104176A (ko) | 2020-11-02 | 2023-07-07 | 네오이뮨텍, 인코퍼레이티드 | 코로나바이러스의 치료를 위한 인터류킨-7의 용도 |
AU2021376396A1 (en) | 2020-11-05 | 2023-06-08 | Neoimmunetech, Inc. | Method of treating a tumor with a combination of an il-7 protein and a nucleotide vaccine |
WO2022117569A1 (en) | 2020-12-02 | 2022-06-09 | Oncurious Nv | A ccr8 antagonist antibody in combination with a lymphotoxin beta receptor agonist antibody in therapy against cancer |
WO2023130081A1 (en) | 2021-12-30 | 2023-07-06 | Neoimmunetech, Inc. | Method of treating a tumor with a combination of il-7 protein and vegf antagonist |
WO2023133595A2 (en) | 2022-01-10 | 2023-07-13 | Sana Biotechnology, Inc. | Methods of ex vivo dosing and administration of lipid particles or viral vectors and related systems and uses |
WO2023193015A1 (en) | 2022-04-01 | 2023-10-05 | Sana Biotechnology, Inc. | Cytokine receptor agonist and viral vector combination therapies |
WO2024102722A1 (en) | 2022-11-07 | 2024-05-16 | Neoimmunetech, Inc. | Methods of treating a tumor with an unmethylated mgmt promoter |
CN117050178B (zh) * | 2023-10-13 | 2024-01-12 | 北京百普赛斯生物科技股份有限公司 | 特异性检测il-7的抗体及应用 |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5714585A (en) * | 1987-10-26 | 1998-02-03 | Sterling Winthrop, Inc. | Antibodies that are immunoreactive with interleukin-7 |
US4965195A (en) * | 1987-10-26 | 1990-10-23 | Immunex Corp. | Interleukin-7 |
US5328988A (en) * | 1987-10-26 | 1994-07-12 | Immunex Corporation | Interleukin-7 |
ZA887773B (en) * | 1987-10-26 | 1989-07-26 | Immunex Corp | Interleukin-7 |
US5728680A (en) * | 1987-12-30 | 1998-03-17 | Cytoven J.V. | Methods for normalizing numbers of lymphocytes |
US5459058A (en) * | 1991-03-28 | 1995-10-17 | Benjamin Rich | Cell culture system |
US5223408A (en) * | 1991-07-11 | 1993-06-29 | Genentech, Inc. | Method for making variant secreted proteins with altered properties |
WO1994022473A1 (en) * | 1993-04-01 | 1994-10-13 | University Of Washington | Use of interleukin 7 to improve vaccine potency |
US5696234A (en) * | 1994-08-01 | 1997-12-09 | Schering Corporation | Muteins of mammalian cytokine interleukin-13 |
DE69838552T2 (de) * | 1997-07-14 | 2008-05-21 | Bolder Biotechnology, Inc., Louisville | Derivate des wachstumshormons und verwandte proteine |
CA2343979C (en) * | 1998-09-21 | 2011-11-15 | Schering Corporation | Human interleukin-b50, therapeutic uses |
WO2001075140A1 (en) * | 2000-03-30 | 2001-10-11 | University Of Connecticut | HYBRID CYTOKINE OF IL-7 AND β-CHAIN OF HEPATOCYTE GROWTH FACTOR |
BRPI0418286A (pt) | 2003-12-30 | 2007-05-02 | Merck Patent Gmbh | proteìnas de fusão de il-7 |
EP1746161A1 (en) | 2005-07-20 | 2007-01-24 | Cytheris | Glycosylated IL-7, preparation and uses |
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DE60334301D1 (de) | 2010-11-04 |
DK1527179T3 (da) | 2011-01-03 |
ZA200501914B (en) | 2005-11-30 |
SI1527179T1 (sl) | 2011-04-29 |
CA2494974C (en) | 2014-07-29 |
ATE482273T1 (de) | 2010-10-15 |
WO2004018681A3 (en) | 2004-04-01 |
JP2014147396A (ja) | 2014-08-21 |
HK1075465A1 (en) | 2005-12-16 |
EP1391513A1 (en) | 2004-02-25 |
CA2494974A1 (en) | 2004-03-04 |
IL166543A (en) | 2012-02-29 |
CY1110994T1 (el) | 2015-06-11 |
JP2005534339A (ja) | 2005-11-17 |
WO2004018681A2 (en) | 2004-03-04 |
EP1527179A2 (en) | 2005-05-04 |
NO20050355L (no) | 2005-05-06 |
ES2353006T3 (es) | 2011-02-24 |
PL213710B1 (pl) | 2013-04-30 |
PL373606A1 (en) | 2005-09-05 |
US7585947B2 (en) | 2009-09-08 |
PT1527179E (pt) | 2010-12-07 |
NO332305B1 (no) | 2012-08-20 |
AU2003250216B2 (en) | 2010-08-05 |
IL166543A0 (en) | 2006-01-15 |
EP1527179B1 (en) | 2010-09-22 |
AU2003250216A1 (en) | 2004-03-11 |
US20050249701A1 (en) | 2005-11-10 |
JP2010115203A (ja) | 2010-05-27 |
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