生体内分解性高分子微粒子を用いた子宮局所におけるドラッグデリバリーシステムの開発

• Project/Area Number: 20659258
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Obstetrics and gynecology
• Research Institution: Osaka University
• Principal Investigator: 木村 正 Osaka University, 医学系研究科, 教授 (90240845)
• Co-Investigator(Kenkyū-buntansha): 筒井 建紀 大阪大学, 医学系研究科, 講師 (00294075) ; 中村 仁美 大阪大学, 医学部附属病院, 特任臨床検査技師 (80467571)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2009: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2008: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: 子宮 / drug delivery system (DDS / カチオン化多糖 / bio-nanocapsule(BNC) / drug delivery system(DDS) / 不妊 / pre-eclampsia

Research Abstract

子宮は特に子宮内膜を標的とした場合、全身投与する事なく、子宮腔内に直接アプローチする事により、局所の標的化が可能な臓器である。本研究では、子宮腔内に直接投与する事ができるような、新たな子宮に対するdrug delivery system(DDS)として、(1)カチオン化多糖および(2)B型肝炎ウイルス(HBV)を利用したウイルス粒子を用いた中空ナノ粒子であるbio-nanocapsule(BNC)を用いて、まずこれらが子宮に対する遺伝子導入の担体として用いる事ができるのか検討し、それぞれの最適化を行った。
子宮内膜においては多くの糖鎖の発現が報告されているものの、妊娠に係る病態および子宮体癌に特異的なものは未だ見つかっていない。そこで、本研究ではカチオン化多糖として、経口薬剤のコーティング剤として既に広く用いられている、デキストランを用いた。スペルミンでカチオン化したデキストランとplasmid DNAのナノオーダーの複合体を作製した。
N/P=0.25、アミノ基導入率5%のデキストラン-DNA複合体において、有効な遺伝子導入効率が得る事ができた。
BNCは酵母を用いて精製され、この粒子はすでに全世界で20年以上も臨床で使用されてきたB型肝炎ワクチンと同様の構造であり、BNCを臨床応用する際の毒性や安全性に対する懸念を著しく低減できる。BNCはHBVがその表面に肝臓に特異的な結合部位を持つために、全身投与したとしても肝臓に特異的に送達される特性を持っている。我々はこの肝臓に特異的な結合部位であるpre-S1領域のN末端をTATペプチドに置換した。TAT-BNCにより、子宮内膜上皮細胞において一過性の有効な遺伝子効率が得る事ができた。今回、TAT修飾したpre-S1領域は糖鎖やサイトカインなどを修飾する事も可能であり、将来病態特異的な物質が明らかになれば、細胞標的化したDDSとしてこれを用いる事ができると考える。
これら結果より、カチオン化デキストランおよびTAT-BNCが子宮に対する新たなDDSに成り得る事が示唆された。

Research Products

• [Journal Article] Concurrent weekly nedaplatin, external beam radiotherapy and high-dose rate brachytherapy in patients with FIGO stage IIIB cervical cancer : A comparison with a cohort treaated by radiotheraphy alone (2010)
  • Author(s): Mabuchi, S., Ugaki, H., Isohashi, F., Yoshioka, Y., Temma, K., Yoda-Hashimoto, N., Takeda, T., Yamamoto, T., Yoshino, K., Nakajima, R., Kuragaki, C., Morishige, K., Enomoto, T., Inoue, T., Kimura, T.
  • Journal Title: Gynecologic and Obstetric Investigation 69 Pages: 224-232
  • Peer Reviewed
• [Journal Article] Temporal and spatial expression of tumor-associated antigen RCASI in pregnant mouse uterus. (2010)
  • Author(s): Tskitishvili, E., Nakamura, H., Kinugasa-Taniguchi, Y., Kanagawa, T., Kimura, T., Tomimatsu, T., Shimoya, K.
  • Journal Title: Int J Gynecol Cancer. 63 Pages: 137-143
  • Peer Reviewed
• [Journal Article] Spinal subarachnoid hematoma following spinal anesthesia in a patient with HELLP syndrome. (2010)
  • Author(s): Koyama, S., Tomimatsu, T, Kanagawa, T., Sawada, K., Tsutsui, T., Kimura, T., Chang, YS., Wasada, K., Imai, S., Murata, Y.
  • Journal Title: Int J Obstet Anesth. 19 Pages: 87-91
  • Peer Reviewed
• [Journal Article] Oxidative stress-induced S100B protein from placenta and amnion affects soluble Endoglin release from endothelial cells. (2010)
  • Author(s): Tskitishvili, E., Sharentuya, N., Temma-Asano, K., Mimura, K., Kinugasa-Taniguchi, Y., Kanagawa, T Fukuda, H., Kimura, T, Tomimatsu, T., Shimoya, K.
  • Journal Title: Mol Hum Reprod 16 Pages: 188-199
  • Peer Reviewed
• [Journal Article] A case of complete hydatidiform mole with co-existent fetus; conclusive diagnosis of androgenesis of the molar placenta by variation of paternal acrocentric short arm (2010)
  • Author(s): Koyama, S., Tomimatsu, T., Sawada, K., Kanagawa, T., Isobe, A., Kinugasa, Y., Tsutsui,T,,Kmura T.
  • Journal Title: America Journal of Perinatology 27 Pages: 143-149
  • Peer Reviewed
• [Journal Article] 妊娠子宮局所におけるマクロファージの役割 (2010)
  • Author(s): 中村仁美、木村正
  • Journal Title: 医学のあゆみ 233巻(In press)
  • Peer Reviewed
• [Journal Article] Spinal subarachnoid hematoma following spinal anesthesia in a patient with HELLP syndrome (2009)
  • Author(s): Koyama, S., Tomimatsu, T., Kanagawa, T., Sawada, K., Tsutsui, T., Kimura, T., Chang YS., Wasada.K., Imai, S., Murata, Y.
  • Journal Title: Int Obestet Anesth 19 Pages: 87-91
  • Peer Reviewed
• [Journal Article] Evidence that oxytocin exerts anxiolytic effects via oxytocin receptor expressed in serotonergic neurons in mice (2009)
  • Author(s): Yoshida, M., Takayanagi, Y., Inoue, K., Kimura, T,, Young, L.J., Onaka, T., Nishimori, K.
  • Journal Title: Journal of Neuroscience 29 Pages: 2259-2271
  • Peer Reviewed
• [Journal Article] Uterine cervical carcinoma : preoperative staging 3.0-T MR imaging-comparison with1.5-TMR imaging (2009)
  • Author(s): Hori, M., Kim, T., Murakami, T., Imaoka, I., Onishi, H., Tomoda, K., Tsutsuui, T., Enomoto, T., Kimura, T., Nakamura, H.
  • Journal Title: Radiology 251 Pages: 96-104
  • Peer Reviewed
• [Journal Article] The activity of carboplatin and paclitaxel for recurrent cervical cancer after definitive radiotherapy (2009)
  • Author(s): Mabuchi, S., Morishige, K., Fujita, M., Tsutsui, T., Sakata, M., Enomoto T., Kimura, T.
  • Journal Title: Gynecologic Oncology 113 Pages: 200-204
  • Peer Reviewed
• [Journal Article] Amniotic fluid `sludge' detected in patients with subchorionic hematoma : a report of two cases (2009)
  • Author(s): Tskitishvili, E,, Tomimatsu, T., Kanagawa, T., Sawada, Y., Kinugasa, Y., Mimura, K., Kimura, T.
  • Journal Title: Ultrasouond Obstet Gynecol 33 Pages: 484-486
  • Peer Reviewed
• [Journal Article] Peripheral quantitative computed tomography is useful to monitor response to alendronate therapy in postmenopausal women (2009)
  • Author(s): Sawada, K., Morishige, K., Nishio, Y., Hayakawa, J., Mabuchi, S., Iosbe, A., Ogata, S., Sakata, M., Ohmichi, M., Kimura, T.
  • Journal Title: J Bone Miner Metab 27 Pages: 175-181
  • NAID: 10025336716
  • Peer Reviewed
• [Journal Article] Cesarean scar pregnancies succseefully treated with methotrexate (2009)
  • Author(s): Muraji, M., Mabuchi, S., Hisamoto, K., Muranishi, M., Kanagawa, T., Nishio, Y., Kimura, T.
  • Journal Title: Acta Obstetricia et Gynecologica Scandinavica 88 Pages: 720-723
  • Peer Reviewed
• [Journal Article] Wilms' tumor I (WTI) peptide immunotherapy for gynecological malignancy (2009)
  • Author(s): Ohno, S., Kyo, S., Myojo, S., Dohi, S., Ishizaki, J., Miyamoto, K-I., Morita, S., Sakamoto, J-I., Enomoto, T., Kimura, T., Oka, Y., Tsuboi, A., Sugiyama, H., Inoue, M.
  • Journal Title: Anticancer Research 29 Pages: 4779-4784
  • Peer Reviewed
• [Journal Article] The activity of carboplatin and paclitaxel for recurrent cervical cancer after definitive radiotherapy (2009)
  • Author(s): Mabuchi, S., Morishige, K., Fujita, M., Tsutsui, T., Sakata, M., Enomoto, T., Kimura, T.
  • Journal Title: Gynecologic Oncology 113 Pages: 200-204
  • Peer Reviewed
• [Journal Article] A case of recurrent cesarean scar pregnancy (2009)
  • Author(s): Mabuchi, S., Kawase, C., Muraji, M., Kanagawa, T., Kimura, T.
  • Journal Title: Acta Obstetricia et Gynecologica Scandinavica 88 Pages: 1059-1060
  • Peer Reviewed
• [Journal Article] Metal transcription factor-I is involved in hypoxia-dependent regulation of placenta growth factor in trophoblast-derived cells (2009)
  • Author(s): Nishimoto, F., Sakata, M., Minekawa, R., Okamoto, Y., Miyake, A., Isobe, A., Yamamoto, T., Takeda, T., Ishida, E., Sawada, K., Morishige, K-I., Kimura, T.
  • Journal Title: Endocrinology 150 Pages: 1801-1808
  • Peer Reviewed
• [Journal Article] Up-regulation of alpha5-integrin by E-cadherin loss in hypoxia and its key role in the migration of extravillous trophoblast cells during early implantation. (2009)
  • Author(s): Arimoto-Ishida, E., Sakata, M., Sawada, K., Nakayama, M., Nishimoto F., Mabuchi, S., Takeda, T., Yamamoto, T., Isobe, A., Okamoto, Y., Lengyel, E., Suehara, N., Morishige, K., Kimura, T.
  • Journal Title: Endocrinology. 150 Pages: 4306-4315
  • Peer Reviewed
• [Journal Article] Calcineurin/NFAT pathway : a novel regulator of parturition (2009)
  • Author(s): Tabata, C., Ogita, K., Sato, K., Nakamura, H., Negoro, H., Kumasawa, K., Temma-Asano, K., Tsutsui, T., Nishimori, K., Kimura, T.
  • Journal Title: American Journal of Reproductive Immunology 62 Pages: 44-50
  • Peer Reviewed
• [Journal Article] MR Imaging of endometrial carcinoma for preoperative staging at 3.O T : comparison with imaging at 1.5 T (2009)
  • Author(s): Hori, M., Kim, T., Murakami, T., Imaoka, I., Onishi, H., Nakamoto, A., Nakaya, Y., Tomoda, K., Tsutsui, T., Enomoto, T., Kimura, T., Nakamura, H. ,
  • Journal Title: Journal of Magnetic Resonance Imaging 30 Pages: 621-630
  • Peer Reviewed
• [Journal Article] mTOR is a promising therapeutic target both in cisplatin-sensitive and cisplatin-resistant clear cell carcinoma of the ovary. (2009)
  • Author(s): Mabuchi, S., Kawase, C., Aitomare, DA., Morishige, K., Sawada, K., Hayashi, M., Tsujimoto, M., Yamoto, M., Klein-Szanto, AJ., Schilder, RJ., Ohmichi, M., Testa, JR., Kimura, T.
  • Journal Title: Clin Cancer Res 15 Pages: 5404-5413
  • Peer Reviewed
• [Journal Article] Postoperative concurrent nedaplatin-based chemoradiotherapy imprves survival in early-stage cervical cancer patients with adverse risk factors. (2009)
  • Author(s): Mabuchi, S., Morishige, K., Isohashi, F., Yoshioka., Y, Takeda, T., Yamamoto, T., Yoshino, K., Enomoto, T, Inoue, T., Kimura, T.
  • Journal Title: Gynecologic Oncology 115 Pages: 482-487
  • Peer Reviewed
• [Journal Article] Postpartum outcome of cervical intraepithelial neoplasia in pregnant women determined determined b route of delivery (2009)
  • Author(s): Ueda, Y., Enomoto, T., Miyatake, T., Yoshino, K., Fujita, M., Miyake, T., Fujiwara, K., Muraji, M., Kanagawa,
  • Journal Title: Reproductive Science 16 Pages: 1034-1039
  • Peer Reviewed
• [Journal Article] Fasudil inhibits lysophosphatidic acid-induced invasiveness of human ovarian cancer cells. (2009)
  • Author(s): Ogata, S., Morishige, K., Sawada, K., Hashimoto, K., Mabuchi, S., Kawase, C., Ooyagi, C., Sakata, M., Kimura, T.
  • Journal Title: Int J Gynecol Cancer. 19 Pages: 1473-1480
  • Peer Reviewed
• [Journal Article] 妊娠子宮局所におけるマクロファージの役割 (2009)
  • Author(s): 中村仁美、香山晋輔、谷口武、荻田和秀、熊澤恵一、張慶、筒井建紀、古山将康、杉野法広、田村博史、金田安史、木村正
  • Journal Title: 生殖内分泌学会雑誌 14巻 Pages: 35-40
• [Journal Article] Late postpartum eclampsia : report of two cases managed by uterinecurettage and review of the literature (2008)
  • Author(s): Matsuo K., Rimura T., et al.
  • Journal Title: Am J Perinatol 24 Pages: 257-266
  • Peer Reviewed
• [Journal Article] Relationship between metabolic syndrome and uterine leiomyomata : a case-control study (2008)
  • Author(s): Takeda T., Kimura T., et al
  • Journal Title: Gynecol Obstet Invest 66 Pages: 14-17
  • Peer Reviewed
• [Journal Article] Dual repressive effect of angiotensin II-type l receptor blockertelmisartan on angiotensin II-induced and estradiol-induceduterine leiomyoma cell proliferation (2008)
  • Author(s): Isobe S., Kimura T., et al
  • Journal Title: Human Reproduction 23 Pages: 440-446
  • Peer Reviewed
• [Journal Article] Infusion of oxytocin induces successful delivery in prostanoid FP-receptor-deficient mice (2008)
  • Author(s): Rawamata M., Kimura K., et al
  • Journal Title: Molecular and Cellular Endocrinology 283 Pages: 32-37
  • Peer Reviewed
• [Journal Article] Maternal urinoma during pregnancy (2008)
  • Author(s): Ushioda N., Kimura T., et al
  • Journal Title: Journal of Obstetrics and Gynaecology Research 34 Pages: 88-91
  • NAID: 10021250356
  • Peer Reviewed
• [Journal Article] Inununisation with a plasmid DNA vaccine encoding gonadotrophinreleasing hormone (GnRH-1) and T-helper epitopes in salinesuppresses rodent fertility (2008)
  • Author(s): Khan M. A. H, Kimura T., et al
  • Journal Title: Vaccine 26 Pages: 1365-1374
  • Peer Reviewed
• [Journal Article] Effect iveness of the herbal medicine Daikenchuto forradiat ion-induced enteritis (2008)
  • Author(s): Takeda T., Kimura T., et al
  • Journal Title: The Journal of Alternative and Complementary Medicine 14 Pages: 753-755
  • Peer Reviewed
• [Journal Article] In vitro and in vivo assays to analyze the contribution of Rhokinase in angiogenesis (2008)
  • Author(s): Sawada K., Kimura T., et al
  • Journal Title: Methods in Enzymology 439 Pages: 395-412
  • Peer Reviewed
• [Journal Article] Frequent inactivation of RUNX3 in endometrial carcinoma (2008)
  • Author(s): Yoshizaki T., Rifflura T. etal.
  • Journal Title: Gynecologic Oncology 110 Pages: 439-444
  • Peer Reviewed
• [Journal Article] PIK3CA gene mutations and amplifications in uterine cancers, identi fied by methods that avoid confounding by PIR3CA pseudogeneseauences (2008)
  • Author(s): Miyake T., Kimura T., et al
  • Journal Title: Cancer letter 261 Pages: 120-126
  • Peer Reviewed
• [Journal Article] The human tumor-associated ant igen RCASI in pregnanciescomplicated by pre-eclampsia (2008)
  • Author(s): Tskitishvili E., Kimura T. etal.
  • Journal Title: Journal of Reproductive Immunology 77 Pages: 100-108
  • Peer Reviewed
• [Journal Article] Maintnance treatment wi th bevacizumab prolongs survival in an invivo ovarian cancer model (2008)
  • Author(s): Mabuchi S., Kimura T., et al
  • Journal Title: Clinical Cancer Research 14 Pages: 7781-7789
  • Peer Reviewed
• [Journal Article] Spatio-temporal image correlation (SITC)used by generalobstetrician : is as effect ive as 2D fetal echocardiography scannedby experts (2008)
  • Author(s): Wani tpongpan T. Rimura T., et al
  • Journal Title: Perinatal Diagnosis 28 Pages: 923-928
  • Peer Reviewed
• [Journal Article] 産婦人科医療体制に発想の転換を一過酷な労働と高い訴訟リスクからの脱却 (2008)
  • Author(s): 木村 正
  • Journal Title: 都市問題 99 Pages: 17-22
  • Peer Reviewed
• [Journal Article] 子宮筋腫に対する子宮動脈塞栓術 (2008)
  • Author(s): 武田 卓, 木村 正, 他
  • Journal Title: 産婦人科治療 97 (2) Pages: 157-159
  • Peer Reviewed
• [Journal Article] 不正出血・過多月経・大量出血に対する対応 (2008)
  • Author(s): 木村 正
  • Journal Title: 救急医学 32 Pages: 1067-1072
  • Peer Reviewed
• [Journal Article] 卵巣癌における血管新生抑制療法 (2008)
  • Author(s): 馬淵誠二, 木村 正, 他
  • Journal Title: 臨床婦人科産科 62 (10) Pages: 1339-1345
  • Peer Reviewed
• [Journal Article] 臨床医の婦人科がん検診-病理標本を見ない実地医科のために- (2008)
  • Author(s): 木村 正
  • Journal Title: 産婦人科治療 97 (3) Pages: 225-231
  • Peer Reviewed
• [Journal Article] 婦人科がんの検診-その効果と問題点- 企画者のことば (2008)
  • Author(s): 木村 正
  • Journal Title: 産婦人科の実際 57 (9) Pages: 1340-1340
  • Peer Reviewed
• [Journal Article] 卵巣がんスクリーニングの有効性と限界 (2008)
  • Author(s): 上田 豊, 木村 正, 他
  • Journal Title: 産婦人科の実際 57 (9) Pages: 1341-1404
  • Peer Reviewed
• [Presentation] Endometrial Stat3 activation and implantation failure (2009)
  • Author(s): Kimura, T.
  • Organizer: IUPS Satellite Symposium on Endometrial Receptivity and Blastocyst Implantation
  • Place of Presentation: lnamori Hall, Kyoto University
  • Year and Date: 2009-07-25
• [Presentation] Mouse model of human infertility : transient and local inhibition of endo metrial STAT-3 activation results in implantation failure (2009)
  • Author(s): Kimura, T.
  • Organizer: 5^<th> lnternational Conference on the Female Reproductive Tact
  • Place of Presentation: Frauenchiemsee, Germany
  • Year and Date: 2009-05-17
• [Presentation] 一過性遺伝子導入方法を用いた着床現象の検討(シンポジウム) (2008)
  • Author(s): 中村仁美, 筒井建紀, 木村 正, 他
  • Organizer: 第23回日本生殖免疫学会
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2008-12-07
• [Presentation] 着床期子宮局所におけるSTAT-3活性の役割 (2008)
  • Author(s): 中村仁美, 筒井建紀, 木村 正, 他
  • Organizer: 第13回日本内分泌学会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2008-11-29
• [Presentation] 着床期子宮内膜におけるマクロファージの役割 (2008)
  • Author(s): 中村仁美, 筒井建紀, 木村 正, 他
  • Organizer: 第53回日本生殖医学会総会・学術講演会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2008-10-24
• [Presentation] 子宮局所における一過性遺伝子導入方法を用いた着床不全マウスモデルの作製 (2008)
  • Author(s): 香山晋輔, 中村仁美, 筒井建紀, 木村 正, 他
  • Organizer: 第53回日本生殖医学会総会・学術講演会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2008-10-24
• [Presentation] 教育講演5「一過性遺伝子導入マウスを用いた妊娠成立期における子宮内膜機能の解析」 (2008)
  • Author(s): 木村 正
  • Organizer: 第53回日本生殖医学会総会・学術講演会
  • Place of Presentation: 神戸国際会議場
• [Presentation] 子宮局所における一過性遺伝子導入方法を用いたヒト不妊症のマウスモデルの作製 (2008)
  • Author(s): 中村仁美, 筒井建紀, 木村 正, 他
  • Organizer: 第2 4回日本DDS学会
  • Place of Presentation: 六本木アカデミーヒルズ
• [Book] スーパーローテイターのための内視鏡外科手術トレーニングマニュアル (2008)
  • Author(s): 木村 正
  • Total Pages: 2
  • Publisher: シュプリンガー・ジャパン
• [Book] 今日の治療指針 (2008)
  • Author(s): 木村 正
  • Total Pages: 2
  • Publisher: 医学書院
• [Remarks] 第13回日本生殖内分泌学会学術奨励賞受賞