JP6087338B2 - 医薬製剤 - Google Patents
医薬製剤 Download PDFInfo
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- JP6087338B2 JP6087338B2 JP2014501706A JP2014501706A JP6087338B2 JP 6087338 B2 JP6087338 B2 JP 6087338B2 JP 2014501706 A JP2014501706 A JP 2014501706A JP 2014501706 A JP2014501706 A JP 2014501706A JP 6087338 B2 JP6087338 B2 JP 6087338B2
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- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/10—Drugs for disorders of the endocrine system of the posterior pituitary hormones, e.g. oxytocin, ADH
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Description
ゴナドトロピンは、雄および雌の生殖腺機能を制御する一群のヘテロ二量体糖タンパク質ホルモンである。それらには卵胞刺激ホルモン(FSH)、黄体形成ホルモン(LH)、および絨毛性ゴナドトロピン(CG)が含まれる。
hCG製剤にはかなりの不均一性が認められ、これは、含有される種々のアイソフォームの量の相違に関係する。個々のhCGアイソフォームは同一のアミノ酸配列を示すが、翻訳後修飾される度合いが異なっている;特定のアイソフォームは糖鎖分枝構造が不均一であること、およびシアル酸(末端糖)の導入量が異なっていることを特徴とし、そのいずれも、特定のアイソフォームの生体活性に影響を与えると考えられる。
(R)n−X−Y (Ia)
式中、(R)はポリエチレングリコール(PEG)またはメトキシポリ(エチレングリコール)(mPEG)であり;nは1、2、3、または4であり;Xは結合またはリンカーであり;そして、YはhCGまたはそのアゴニスト変異体である。好ましくは、nが1であってXが共有結合であるか、または、nが1、2、もしくは4であってXがリンカーである。好ましくは、リンカーXの一末端がhCGのN-末端に結合している。
(R)n−X−Y (I)
式中、
Rは水溶性で実質的に非抗原性のポリマーであり;
nは1、2、3、または4であり;
Xは結合またはリンカーであり;そして、
YはhCGまたはそのアゴニスト変異体である。
ヒトhCG
FiddesおよびGoodman (1979)に従い、hCGαポリペプチド遺伝子のコード領域を使用した。配列はAH007338として登録されており、構築の時点で、このタンパク質配列には他の変異体は存在しなかった。本明細書ではこの配列をSEQ ID 1と称する。
シアリルトランスフェラーゼ
hCGαポリペプチドのコード配列(AH007338, SEQ ID 1)およびhCGβポリペプチドのコード配列(NP_000728, SEQ ID 2)をPCRで増幅した(プライマーはそれぞれ、CGa-fwおよびCGa-rev、CGb-fwおよびCGb-revの組み合わせで使用)。
βガラクトシドα-2,3-シアリルトランスフェラーゼ4のコード配列(ST3、L23767、SEQ ID 3)のPCR増幅を、プライマー、2,3STfwおよび2,3STrevを使用して行った。
βガラクトサミドα-2,6-シアリルトランスフェラーゼ1のコード配列(ST6、NM_003032、SEQ ID 4)のPCR増幅を、プライマー、2,6STfwおよび2,6STrevを使用して行った。
クローンのトランスフェクション、単離、およびスクリーニング
hCGを生成するPer.C6クローンを作製するために、1つのプラスミドからhCGの両ポリペプチド鎖を発現させた(実施例1参照)。
高度にシアリル化されたhCGを生成するPer.C6クローンを作製するために、既にhCGの両ポリペプチド鎖を発現することが確認されているPer.C6細胞(実施例4参照)において、別のプラスミド(実施例2参照)からα2,3-シアリルトランスフェラーゼを発現させた。実施例4に記載するPER.C6(登録商標)細胞から作製したクローンについて、その特性(生産性、良好な増殖プロファイル、機能性タンパク質の生成、および生成されるシアリル化含有hCG)に基づく選択を行った。
上記のように作製したα、βヘテロダイマー(実施例4)はシアリル化のレベルが低く、非常に塩基性の高いIEFプロファイルを示した。上記(実施例5a)のように、Per.C6細胞におけるhCGおよびα2,3-シアリルトランスフェラーゼの共発現により、hCGのみを発現する細胞に比較して、高レベルのシアリル化hCGが得られる。
電気泳動は、電場による溶媒中での荷電分子の移動と定義される。電場による生体分子の移動度は電場の強度、分子の正味荷電、分子のサイズおよび形状、分子が移動する溶媒のイオン強度および特性に依存する。
レクチンに基づくグリカン識別法を用いて、複合糖質の分析を行った。この方法によれば、ニトロセルロースに結合した糖タンパク質および複合糖質のキャラクタリゼーションを行うことができる。レクチンは特定の部分(例えばα2,3結合型シアル酸)を選択的に認識する。適用したレクチンはステロイド・ハプテンであるジゴキシゲニンとコンジュゲートし、これによって結合したレクチンの免疫学的検出が可能となる。
シアル酸はタンパク質に結合した炭水化物であってモノサッカライドと見なされ、他のモノサッカライド(例えばガラクトース、マンノース、グルコサミン、ガラクトサミン、およびフコース)と化合して存在する。本発明の精製rhCG上の総シアル酸を、Stantonらの方法に基づく方法を用いて測定した(J. Biochem. Biophys. Methods. 30 (1995), 37 - 48)。
α2,3-シアリルトランスフェラーゼで改変したPer.C6組換えhCG(例えば実施例5a、実施例5b)の総シアル酸含量を測定したところ、(タンパク質のモル数に対するシアル酸のモル数の比で)15 mol/mol以上、例えば18 mol/mol以上、例えば19.1 mol/molであった。これはOvitrelle(総シアル酸含量 17.6 mol/mol)に匹敵する。
α2,3-シアリルトランスフェラーゼで改変したPer.C6組換えhCG(080019-19)(上記実施例5bの方法で調製したもの)の総シアル酸含量を測定したところ、(タンパク質のモル数に対するシアル酸のモル数の比で)20 mol/molであった。この場合も、これはOvitrelle(総シアル酸含量 17.6 mol/mol)を凌ぐものである。この事例(080019-19)について試験を行い、α2,3およびα2,6シアル酸の相対量を定量した(実施例8C)。
精製rhCG((080019-19)の例および実施例5で調製した他の2つの例)上のα2,3およびα2,6シアル酸の相対量(%)を、既知の方法、すなわち順相(NP-)HPLCを用いて測定した。
精製rhCG(実施例8Cで使用した3つのサンプル)から抽出したグリカン上のモノ-、ジ-、トリ-、およびテトラ-シアリル化構造の相対量(%)を既知の方法で測定した。
α2,3-シアリルトランスフェラーゼを用いて操作したPer.C6組換えhCGサンプル(例えば実施例5a、5b)の代謝クリアランス速度(MCR)を測定するために、意識のあるメス・ラット(クローン毎に3検体)の尾静脈にrhCGをボーラス注射し(サンプルのELISA定量(DRG社、EIA 1288)に基づき、1-10 μg/ラット)、時間=0とした。試験サンプル注射の1、2、4、8、12、24、および32時間後に、尾の先端部から血液サンプル(400μL)を採取した。遠心分離によって血清を回収し、ELISA(DRG社、EIA 1288)によってhCG含量のアッセイを行った。α2,3-シアリルトランスフェラーゼで操作したPer.C6 hCGサンプルのMCRは、半減期が標準物質と同程度であることを示した(図5)。図6は、α2,3-シアリルトランスフェラーゼで操作した他のhCGサンプルの半減期が標準物質と比較して向上されることを示している(図6)。
hCGバイオアッセイを実施し、hCG特異的活性を測定した。活性の測定はUSP(USP Monographs:Chorionic Gonadotropin, USPC Official 8/1/09-11/30/09)に従い、Ovitrelleを標準物質として用いて行った。Ovitrelleの生体活性は26,000 IU/mgである(Curr Med Res Opin. 2005 Dec; 21(12): 1969 - 76)。許容限界(acceptance limit)は>21,000 IU hCG/mgである。α2,3-シアリルトランスフェラーゼで操作したヒト細胞株由来組換えhCGサンプル(シアル酸含量 19.1 mol/mol、実施例8参照)の生体活性は27,477 IU hCG/mgであった。
無血清培地中で懸濁培養したPER.C6細胞において組換えhCGを生成させる方法を開発した。方法は以下に記載するが、いくつかのhCG生成PER.C6細胞株に適用される。
全てのクロマトグラフィー操作で、免疫反応性組換えhCGの存在をRIA(DRG社、EIA 1288)およびIEF(実施例6)で確認した。
本発明のある例では、ポリ(エチレングリコール)はhCG(またはそのアゴニスト変異体)のアミノ酸残基を介して(例えば共有)結合している。好ましくは、ポリ(エチレングリコール)はhCGのN-末端に結合している。種々の官能基、リンカー、立体配置、および分子量を有する多くの活性ポリ(エチレングリコール)が当業者に知られている。それらを使用して、当該分野で知られる方法によってPEG-hCGコンジュゲートまたはPEG-hCGアゴニスト変異体コンジュゲートを合成してもよい(例えばRoberts M. J.ら, Adv. Drug Del. Rev. 54: 459-476,2002;Harris J. M.ら, Drug Delivery Sytems 40: 538-551,2001参照)。
m-PEG-アルデヒドの官能基は、pH5.5で主にN-末端と相互作用する。直鎖m-PEG-アルデヒドを用いて行う実験では、期待される2分枝ではなく約6から7分枝のPEG鎖を有するPEG化hCGが生成される。これは、m-PEG-アルデヒドが他のアミノ酸、例えばヒスチジン(hCGは4個のヒスチジン残基を有する)と相互作用することを示唆している。
rhCG(研究室内で実施例5b記載の方法によって生成し、実施例11記載の方法によって精製したもの)を、実施例12Aの方法を用いて直鎖m-PEG-アルデヒド(10KD)で修飾した。実施した手順はスキーム3および以下の通りである:
組換えhCGを実施例5bの方法によって生成し、実施例11記載の方法によって精製した。21検体のラットを3群に分け(7検体/群)、各ラットに以下のいずれかの用量の組換えhCGを、3つの独立した日に3回注射した(各注射は異なる日に行った)(1群=1用量):4.3ng(A群のラット)、8.6ng(B群のラット)、および17.1ng(C群のラット)。5日後、ラットを殺処分し、既知の慣例的なバイオアッセイ法に従って、効力測定のために子宮の重量を測定した。
(i)研究室内で実施例5bの方法によって生成し、実施例11の方法によって精製した組換えhCGの単回注射。一日一回注射を3回行った際の最も高い濃度より10倍高い濃度(170ng)で投与;
(ii)実施例12Aの方法によって生成した本発明の分枝PEG化組換えhCGの単回注射。一日一回注射を3回行った際の最も高い濃度より10倍高い濃度(170ng)で投与;および、
(iii)実施例12の方法によって生成した本発明の直鎖PEG化組換えhCGの単回注射。一日一回注射を3回行った際の最も高い濃度より10倍高い濃度(170ng)で投与。
結果を以下の表に示す。
Andersen CY, Westergaard LG, and van Wely M. (2004). FSH isoform composition of commercial gonadotrophin preparations: a neglected aspect? Reprod Biomed Online. 9(2), 231-236.
Bassett RM, and Driebergen R. (2005). Continued improvements in the quality and consistency of follitropin alfa, recombinant human FSH. Reprod Biomed Online. 10(2), 169-177.
D'Antonio M., Borrelli F. , Datola A., Bucci R. , Mascia M. , Polletta P., Piscitelli D., and Papoian R. (1999) Biological characterization of recombinant human follicle stimulating hormone isoforms. Human Reproduction 14, 1160-1167
Fiddes, J. C. and Goodman, H. M. (1979) Isolation, cloning and sequence analysis of the cDNA for the alpha-subunit of human chorionic gonadotropin. Nature, 281, 351-356.
Fiddes, J. C. and Goodman, H. M. (1980) The cDNA for the beta-subunit of human chorionic gonadotropin suggests evolution of a gene by readthrough into the 3'-untranslated region. Nature, 286, 684-387.
Kagawa Y, Takasaki S, Utsumi J, Hosoi K, Shimizu H, Kochibe N, and Kobata A. (1988). Comparative study of the asparagine-linked sugar chains of natural human interferon-beta 1 and recombinant human interferon-beta 1 produced by three different mammalian cells. J Biol Chem. 263(33), 17508-17515.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. (1951) Protein measurement with the Folin phenol reagent. J Biol Chem. 193(1), 265-75.
Lowry, PJ, McLean, C, Jones RL and Satgunasingam N. (1976) Purification of anterior pituitary and hypothalamic hormones Clin Pathol Suppl (Assoc Clin Pathol). 7, 16-21.
Royle L, Radcliffe CM, Dwek RA and Rudd PM (2006) Methods in Molecular Biology, ed I Brockhausen-Schutzbach (Humana Press), 347: Glycobiology protocols, 125-144.
Steelman SL, and Pohley FM. (1953) Assay of the follicle stimulating hormone based on the augmentation with human chorionic gonadotropin. Endocrinology. 53(6), 604-616.
Svensson EC, Soreghan B, and Paulson JC. (1990) Organization of the beta-galactoside alpha 2,6-sialyltransferase gene. Evidence for the transcriptional regulation of terminal glycosylation. J Biol Chem. 265(34):20863-20868.
Takeuchi M, Takasaki S, Miyazaki H, Kato T, Hoshi S, Kochibe N, and Kobata A (1988). Comparative study of the asparagine-linked sugar chains of human erythropoietins purified from urine and the culture medium of recombinant Chinese hamster ovary cells. J Biol Chem. 263(8), 3657-3663.
Ulloa-Aguirre A, Midgley AR Jr, Beitins IZ, and Padmanabhan V. (1995). Follicle-stimulating isohormones: characterization and physiological relevance. Endocr Rev.16(6), 765-787.
Ulloa-Aguirre A, Timossi C, Barrios-de-Tomasi J, Maldonado A, and Nayudu P. (2003). Impact of carbohydrate heterogeneity in function of follicle-stimulating hormone: studies derived from in vitro and in vivo models. Biol Reprod. 69(2), 379-389.
ヒト絨毛性ゴナドトロピンアルファポリペプチド
寄託番号AH007338
hCGアルファのヌクレオチド配列
1 ATGGATTACT ACAGAAAATA TGCAGCTATC TTTCTGGTCA CATTGTCGGT GTTTCTGCAT
61 GTTCTCCATT CCGCTCCTGA TGTGCAGGAT TGCCCAGAAT GCACGCTACA GGAAAACCCA
121 TTCTTCTCCC AGCCGGGTGC CCCAATACTT CAGTGCATGG GCTGCTGCTT CTCTAGAGCA
181 TATCCCACTC CACTAAGGTC CAAGAAGACG ATGTTGGTCC AAAAGAACGT CACCTCAGAG
241 TCCACTTGCT GTGTAGCTAA ATCATATAAC AGGGTCACAG TAATGGGGGG TTTCAAAGTG
301 GAGAACCACA CGGCGTGCCA CTGCAGTACT TGTTATTATC ACAAATCTTA A
1 MDYYRKYAAI FLVTLSVFLHVLHSAPDVQDCPECTLQENPFFSQPGAPILQCMGCCFSRA
61 YPTPLRSKKTMLVQKNVTSESTCCVAKSYNRVTVMGGFKVENHTACHCSTCYYHKS
ヒト絨毛性ゴナドトロピンベータポリペプチド
寄託番号NP_000728
hCGベータのヌクレオチド配列
ヌクレオチド配列
1 ATGGAGATGT TCCAGGGGCT GCTGCTGTTG CTGCTGCTGA GCATGGGCGG GACATGGGCA
61 TCCAAGGAGC CGCTTCGGCC ACGGTGCCGC CCCATCAATG CCACCCTGGC TGTGGAGAAG
121 GAGGGCTGCC CCGTGTGCAT CACCGTCAAC ACCACCATCT GTGCCGGCTA CTGCCCCACC
181 ATGACCCGCG TGCTGCAGGG GGTCCTGCCG GCCCTGCCTC AGGTGGTGTG CAACTACCGC
241 GATGTGCGCT TCGAGTCCAT CCGGCTCCCT GGCTGCCCGC GCGGCGTGAA CCCCGTGGTC
301 TCCTACGCCG TGGCTCTCAG CTGTCAATGT GCACTCTGCC GCCGCAGCAC CACTGACTGC
361 GGGGGTCCCA AGGACCACCC CTTGACCTGT GATGACCCCC GCTTCCAGGA CTCCTCTTCC
421 TCAAAGGCCC CTCCCCCCAG CCTTCCAAGT CCATCCCGAC TCCCGGGGCC CTCGGACACC
481 CCGATCCTCC CACAATAA
1 MEMFQGLLLL LLLSMGGTWA SKEPLRPRCR PINATLAVEK EGCPVCITVN TTICAGYCPT
61 MTRVLQGVLP ALPQVVCNYR DVRFESIRLP GCPRGVNPVV SYAVALSCQC ALCRRSTTDC
121 GGPKDHPLTC DDPRFQDSSS SKAPPPSLPS PSRLPGPSDT PILPQ
β-ガラクトシドα-2,3-シアリルトランスフェラーゼ4
寄託番号L23767
ST3GAL4のヌクレオチド配列
1 ATGTGTCCTG CAGGCTGGAA GCTCCTGGCC ATGTTGGCTC TGGTCCTGGT CGTCATGGTG
61 TGGTATTCCA TCTCCCGGGA AGACAGGTAC ATCGAGCTTT TTTATTTTCC CATCCCAGAG
121 AAGAAGGAGC CGTGCCTCCA GGGTGAGGCA GAGAGCAAGG CCTCTAAGCT CTTTGGCAAC
181 TACTCCCGGG ATCAGCCCAT CTTCCTGCGG CTTGAGGATT ATTTCTGGGT CAAGACGCCA
241 TCTGCTTACG AGCTGCCCTA TGGGACCAAG GGGAGTGAGG ATCTGCTCCT CCGGGTGCTA
301 GCCATCACCA GCTCCTCCAT CCCCAAGAAC ATCCAGAGCC TCAGGTGCCG CCGCTGTGTG
361 GTCGTGGGGA ACGGGCACCG GCTGCGGAAC AGCTCACTGG GAGATGCCAT CAACAAGTAC
421 GATGTGGTCA TCAGATTGAA CAATGCCCCA GTGGCTGGCT ATGAGGGTGA CGTGGGCTCC
481 AAGACCACCA TGCGTCTCTT CTACCCTGAA TCTGCCCACT TCGACCCCAA AGTAGAAAAC
541 AACCCAGACA CACTCCTCGT CCTGGTAGCT TTCAAGGCAA TGGACTTCCA CTGGATTGAG
601 ACCATCCTGA GTGATAAGAA GCGGGTGCGA AAGGGTTTCT GGAAACAGCC TCCCCTCATC
661 TGGGATGTCA ATCCTAAACA GATTCGGATT CTCAACCCCT TCTTCATGGA GATTGCAGCT
721 GACAAACTGC TGAGCCTGCC AATGCAACAG CCACGGAAGA TTAAGCAGAA GCCCACCACG
781 GGCCTGTTGG CCATCACGCT GGCCCTCCAC CTCTGTGACT TGGTGCACAT TGCCGGCTTT
841 GGCTACCCAG ACGCCTACAA CAAGAAGCAG ACCATTCACT ACTATGAGCA GATCACGCTC
901 AAGTCCATGG CGGGGTCAGG CCATAATGTC TCCCAAGAGG CCCTGGCCAT TAAGCGGATG
961 CTGGAGATGG GAGCTATCAA GAACCTCACG TCCTTCTGA
1 MCPAGWKLLA MLALVLVVMV WYSISREDRY IELFYFPIPE KKEPCLQGEA ESKASKLFGN
61 YSRDQPIFLR LEDYFWVKTP SAYELPYGTK GSEDLLLRVL AITSSSIPKN IQSLRCRRCV
121 VVGNGHRLRN SSLGDAINKY DVVIRLNNAP VAGYEGDVGS KTTMRLFYPE SAHFDPKVEN
181 NPDTLLVLVA FKAMDFHWIE TILSDKKRVR KGFWKQPPLI WDVNPKQIRI LNPFFMEIAA
241 DKLLSLPMQQ PRKIKQKPTT GLLAITLALH LCDLVHIAGF GYPDAYNKKQ TIHYYEQITL
301 KSMAGSGHNV SQEALAIKRM LEMGAIKNLT SF
β-ガラクトサミドα-2,6-シアリルトランスフェラーゼ1
寄託番号NM_003032
ST6GAL1のヌクレオチド配列
1 ATGATTCACA CCAACCTGAA GAAAAAGTTC AGCTGCTGCG TCCTGGTCTT TCTTCTGTTT
61 GCAGTCATCT GTGTGTGGAA GGAAAAGAAG AAAGGGAGTT ACTATGATTC CTTTAAATTG
121 CAAACCAAGG AATTCCAGGT GTTAAAGAGT CTGGGGAAAT TGGCCATGGG GTCTGATTCC
181 CAGTCTGTAT CCTCAAGCAG CACCCAGGAC CCCCACAGGG GCCGCCAGAC CCTCGGCAGT
241 CTCAGAGGCC TAGCCAAGGC CAAACCAGAG GCCTCCTTCC AGGTGTGGAA CAAGGACAGC
301 TCTTCCAAAA ACCTTATCCC TAGGCTGCAA AAGATCTGGA AGAATTACCT AAGCATGAAC
361 AAGTACAAAG TGTCCTACAA GGGGCCAGGA CCAGGCATCA AGTTCAGTGC AGAGGCCCTG
421 CGCTGCCACC TCCGGGACCA TGTGAATGTA TCCATGGTAG AGGTCACAGA TTTTCCCTTC
481 AATACCTCTG AATGGGAGGG TTATCTGCCC AAGGAGAGCA TTAGGACCAA GGCTGGGCCT
541 TGGGGCAGGT GTGCTGTTGT GTCGTCAGCG GGATCTCTGA AGTCCTCCCA ACTAGGCAGA
601 GAAATCGATG ATCATGACGC AGTCCTGAGG TTTAATGGGG CACCCACAGC CAACTTCCAA
661 CAAGATGTGG GCACAAAAAC TACCATTCGC CTGATGAACT CTCAGTTGGT TACCACAGAG
721 AAGCGCTTCC TCAAAGACAG TTTGTACAAT GAAGGAATCC TAATTGTATG GGACCCATCT
781 GTATACCACT CAGATATCCC AAAGTGGTAC CAGAATCCGG ATTATAATTT CTTTAACAAC
841 TACAAGACTT ATCGTAAGCT GCACCCCAAT CAGCCCTTTT ACATCCTCAA GCCCCAGATG
901 CCTTGGGAGC TATGGGACAT TCTTCAAGAA ATCTCCCCAG AAGAGATTCA GCCAAACCCC
961 CCATCCTCTG GGATGCTTGG TATCATCATC ATGATGACGC TGTGTGACCA GGTGGATATT
1021 TATGAGTTCC TCCCATCCAA GCGCAAGACT GACGTGTGCT ACTACTACCA GAAGTTCTTC
1081 GATAGTGCCT GCACGATGGG TGCCTACCAC CCGCTGCTCT ATGAGAAGAA TTTGGTGAAG
1141 CATCTCAACC AGGGCACAGA TGAGGACATC TACCTGCTTG GAAAAGCCACACTGCCTGGC
1201 TTCCGGACCA TTCACTGCTA A
1 MIHTNLKKKF SCCVLVFLLF AVICVWKEKK KGSYYDSFKL QTKEFQVLKS LGKLAMGSDS
61 QSVSSSSTQD PHRGRQTLGS LRGLAKAKPE ASFQVWNKDS SSKNLIPRLQ KIWKNYLSMN
121 KYKVSYKGPG PGIKFSAEAL RCHLRDHVNV SMVEVTDFPF NTSEWEGYLP KESIRTKAGP
181 WGRCAVVSSA GSLKSSQLGR EIDDHDAVLR FNGAPTANFQ QDVGTKTTIR LMNSQLVTTE
241 KRFLKDSLYN EGILIVWDPS VYHSDIPKWY QNPDYNFFNN YKTYRKLHPN QPFYILKPQM
301 PWELWDILQE ISPEEIQPNP PSSGMLGIII MMTLCDQVDI YEFLPSKRKT DVCYYYQKFF
361 DSACTMGAYH PLLYEKNLVK HLNQGTDEDI YLLGKATLPG FRTIHC
Claims (15)
- ポリ(エチレングリコール)修飾されたhCGであって、式(Ia)を有するhCG:
(R)n−X−Y (Ia)
{式中:
Rがポリエチレングリコール(PEG)またはメトキシポリ(エチレングリコール)(mPEG)であり;
nが2、3、または4であり;
Xがリンカーであり;そして、
YがhCGまたはそのアゴニスト変異体である}。 - nが2または4であってXがリンカーである、請求項1に記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGが組換えhCGである、請求項1または2に記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGがヒト細胞株由来の組換えhCGである、請求項1〜3のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGがα2,3−およびα2,6−シアリル化を含有する組換えhCGである、請求項1〜4のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGが、15mol/molまたはそれ以上のシアル酸含量(タンパク質のモル数に対するシアル酸のモル数の比で表される)を有する組換えhCGである、例えば15mol/molから25mol/molのシアル酸含量を有する組換えhCGである、請求項1〜5のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGが、総シアリル化の10%もしくはそれ以上のα2,3−シアリル化、および/または総シアリル化の50%もしくはそれ未満がα2,6−シアリル化を有する組換えhCGである、請求項1〜6のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGが、総シアリル化の45%から80%のα2,3−シアリル化を有する組換えhCGである、請求項1〜7のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGが、総シアリル化の20%から55%のα2,6−シアリル化を有する組換えhCGである、請求項1〜8のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGのアミノ酸残基にポリ(エチレングリコール)またはメトキシポリ(エチレングリコール)(mPEG)または水溶性で実質的に非抗原性であるポリマーがコンジュゲートしている、請求項1〜9のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- hCGのN−アミノ末端またはC−アミノ末端にポリ(エチレングリコール)、メトキシポリ(エチレングリコール)(mPEG)または水溶性で実質的に非抗原性であるポリマーがコンジュゲートしている、請求項1〜9のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- ポリ(エチレングリコール)またはメトキシポリ(エチレングリコール)(mPEG)または水溶性で実質的に非抗原性であるポリマーの90%またはそれ以上がhCGのN−アミノ末端にコンジュゲートしている、請求項1〜9のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG。
- 請求項1〜12のいずれかに記載されるポリ(エチレングリコール)修飾されたhCGを含む、医薬製剤。
- 請求項1〜12のいずれかに記載されるポリ(エチレングリコール)修飾されたhCG、または請求項13に記載される医薬製剤を含み、FSHおよび/またはLHを更に含んでいてもよい、医薬組成物。
- 不妊症治療用である、請求項14に記載の医薬組成物。
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TWI604850B (zh) * | 2009-10-05 | 2017-11-11 | 菲瑞茵國際中心股份有限公司 | 藥學製劑 |
BR102016006222A2 (pt) * | 2016-03-22 | 2017-09-26 | Universidade De São Paulo - Usp | Process of production and purification of recombinant hydrogen hydrogen hybrid or non-hybrid, recombinant hydrogen hydrogen hybrids or non-hybrid, vectors of expression, and uses of recombinant glicoprotetic hormones |
Family Cites Families (20)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4179337A (en) * | 1973-07-20 | 1979-12-18 | Davis Frank F | Non-immunogenic polypeptides |
US4421896A (en) * | 1979-11-13 | 1983-12-20 | The Dow Chemical Company | Method of coupling a protein to a polymer particle containing hydrazide groups in a polymer latex and the products formed therefrom |
DK49987A (da) | 1987-01-30 | 1988-07-31 | Nordisk Gentofte | Fremgangsmaade til behandling af infertilitet og middel til anvendelse ved fremgangsmaaden |
IT1206302B (it) | 1987-06-26 | 1989-04-14 | Serono Cesare Ist Ricerca | Ormone follicolo-stimolante urinario |
TW518235B (en) | 1997-01-15 | 2003-01-21 | Akzo Nobel Nv | A gonadotropin-containing pharmaceutical composition with improved stability on prolong storage |
CN1302010C (zh) | 1997-06-25 | 2007-02-28 | 应用研究系统Ars股份公司 | 二硫键交联的糖蛋白激素类似物及其制备和应用 |
US6500627B1 (en) | 1998-02-03 | 2002-12-31 | The Trustees Of Columbia University In The City Of New York | Methods for predicting pregnancy outcome in a subject by HCG assay |
EP1257564B1 (en) | 2000-02-22 | 2008-08-27 | Laboratoires Serono SA | Process for purification of recombinant hcg having a specific bioactivity |
MXPA04002256A (es) | 2001-09-12 | 2004-07-23 | Applied Research Systems | Uso de gonadotropina corionica humana (gch) en hiperestimulacion ovarica controlada. |
MEP38608A (en) | 2001-10-22 | 2011-02-10 | Merck Serono Sa | Gonadotrophins for folliculogenesis |
BR0213402A (pt) | 2001-10-29 | 2004-10-13 | Crucell Holland Bv | Métodos para identificar uma célula de mamìfero capaz de produzir uma molécula proteinácea, para selecionar uma célula de mamìfero capaz de produzir uma molécula proteinácea, para se obter uma célula de mamìfero a partir de uma população heterogênea de células, e para produzir uma molécula proteinácea, composição farmaceuticamente aceitável, eritropoietina recombinantemente produzida, usos de uma célula de mamìfero, da eritropoietina recombinantemente produzida, e de uma composição de moléculas semelhantes à eritropoietina, preparação farmacêutica, método para o tratamento preventivo e/ou terapêutico de um distúrbio, composição de moléculas semelhates à eritropoietina, métodos para produzir, em uma célula de mamìfero, moléculas proteináceas, para produzir uma fração enriquecida em uma molécula proteinácea, e para fracionar uma mistura que contém moléculas proteináceas, fração, e, usos de uma fração ou composição, e de eritropoietina recombinantemente produzìvel em uma célula de mamìfero |
US20040248784A1 (en) * | 2003-06-03 | 2004-12-09 | Marco Filicori | Unitary combinations of FSH and hCG |
ATE476666T1 (de) | 2004-02-04 | 2010-08-15 | Centre Nat Rech Scient | Verfahren zur identifizierung von glykoformspezifischen antikörpern |
US8609370B2 (en) | 2004-02-13 | 2013-12-17 | Glycotope Gmbh | Highly active glycoproteins-process conditions and an efficient method for their production |
US20080242607A1 (en) * | 2006-07-21 | 2008-10-02 | Neose Technologies, Inc. | Glycosylation of peptides via o-linked glycosylation sequences |
SI2068907T1 (en) * | 2006-10-04 | 2018-01-31 | Novo Nordisk A/S | Pegylated sugars and glycopeptides are associated with glycerol |
EP2242505A4 (en) * | 2008-01-08 | 2012-03-07 | Biogenerix Ag | GLYCOCONJUGATION OF POLYPEPTIDES USING OLIGOSACCHARYLTRANSFERASES |
TWI488640B (zh) * | 2008-04-16 | 2015-06-21 | Ferring Int Ct Sa | 藥學製劑 |
TWI604850B (zh) | 2009-10-05 | 2017-11-11 | 菲瑞茵國際中心股份有限公司 | 藥學製劑 |
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