201244740 六、發明說明： 【發明所屬之技術領域】 本發明是有關於一種水溶性超順磁性群集奈米粒子之 製造方法，特別是有關於一種水溶性超順磁性氧化鐵群集 奈米粒子之製造方法及其於臨床檢測、診斷、治療之應用。 【先前技術】 磁共振造影為一種非侵入性和非幅射游離性的造影技 術，其在軟組織有良好的成像對比度、較高的空間分辨率， 並有斷層成像的能力，是近年來臨床診斷上相當重要的影 像工具。 現今臨床常使用的顯影劑係以釓類顯影劑為主，釓對 細胞具有毒性，若劑量使用不當或劑型設計不佳，容易造 成嚴重的副作用而影響人體健康。而且，使用釓類顯影劑 有時會出現「偽訊號」的現象，或是因體内分泌影響而稀 釋其濃度造成訊號消失等缺點。因此，其他顯影劑的發展 與應用遂成為磁共振造影技術的重要課題。 目前主要發展的磁共振造影顯影劑之一為超順磁性氧 化鐵（superparamagnetic iron oxide ; SPIO)奈米粒子。在臨 床實驗中，可應用於分子影像、基因標記、非侵入性的細 胞標記或腫瘤偵測等，其應用範圍相當廣泛。 已知的超順磁性氧化鐵奈米粒子製程是在有機相中產 生均勻分佈奈米粒子，而在水溶液相中則多以群集型為 主。雖然可以藉由加入高分子或界面活性劑在粒子表面形 成保護層，以於水溶液中得到較分散的奈米粒子，但因粒 201244740 子表面受到高分子或界面活性劑分子保護而不易與 lit作進一步的應用。再者，未經修飾或包覆保護的； 散，將限制其在生物體中的應用。然而，絕大部分m =為t溶性物質，倘若要將此種奈米粒子運用於3 是非常重要之環節。 船之不未粒子之技術將 【發明内容】 群集樣ί在提供-種水溶性超順-性 奈米粒子與水溶數個超順磁性氧化鐵 形成混合溶液或水溶液， 乳化溶液，其中乳化波震U洛液’使其成為 的超順磁性氧化鐵太複數個小油滴，這些小油滴 維生素E中。舞义不子係呈分散排列且包覆於水溶性 使這此小油滴：：、加熱去除乳化溶液的油相溶劑’以 其中每 劑。本發月之一實施方式，其中油相溶劑為脂肪族溶 酸乙_旨、式’其中脂肪族溶劑包含乙 双夭南、二氣曱烷或正己烷。 依.、、、本實施方式之一實施例，其中脂昉族溶劑包含正 201244740 己烧。 依照本發明之一實施例，水溶液包括三乙胺溶液、阿 黴素水溶液或上述之任意組合。 依照本發明之一實施例，其中正己烷與去離子水或水 溶液之體積比為1 : 50至1 : 1000。 依照本發明之一實施例，其中正己烷與去離子水或水 溶液之體積比為1 : 500。 依照本發明之一實施例，其中超音波震盈5分鐘至15 分鐘。 依照本發明之一實施例，其中加熱上述之混合溶液之 溫度為65°C至105°C。 依照本發明之一實施例，其中上述之混合溶液於75°C 加熱1小時。 依照本發明之一實施例，其中上述之水溶性的群集奈 米粒子之多分散性指數為0.2至0.5。 本發明之又一態樣是在提供一種水溶性超順磁性群集 奈米粒子，其係由上述方法所製得。 本發明之再一態樣是在提供一種磁共振造影顯影劑， 其係包含由上述方法所製成之水溶性超順磁性群集奈米粒 子以及水。 本發明之另一態樣是在提供一種醫藥組成物，其係包 含由以上任一製造方式和實施例所製成之水溶性超順磁性 群集奈米粒子以及一活性物質，且此活性物質係包覆於水 溶性超順磁性群集奈米粒子中。 依照本發明之一實施例，其中上述之活性物質為阿黴 201244740 素。 發溶劑揮 素五中，使其成為w 子匕覆在水溶性維生 列的超順磁性氧化鐵二::奈:粒子’並使原本分散排 此水溶性超順雜錢麟狀群集粒子。 時，可得到較佳之二=粒工應用於磁共振造影顯影劑 群集奈米粒子亦可作=::=== =例如疏水性_)包覆在水紐賴魏群集奈米Lt 【實施方式】 在本發明中’利用水溶性維生素E將超順磁性氧化鎧太 米粒子和活性物質（例如疏水性藥物）包覆在内，而使^ 溶性之超順磁性奈米粒子形成生體安全且為水溶性之超順 磁性群集奈米粒子，其可穩定均勻溶於生理緩衝溶液或其 他水溶液中，進而可應用於臨床檢測、診斷及治療中。 在奈米粒子中，改變表面化學特性會影響奈米粒子的 特性，例如：改變物理與化學性質，包括大小、溶解度、 分散狀態和磁化值等。因此，透過改變表面化學特性可使 磁性奈米粒子在生物系統中的性質產生變化，包括細胞辨 識、生物分佈情形和免疫反應的產生，用以減少奈米粒子 潛在的奈米毒性。201244740 VI. Description of the Invention: [Technical Field] The present invention relates to a method for producing a water-soluble superparamagnetic cluster nanoparticle, and more particularly to the manufacture of a water-soluble superparamagnetic iron oxide cluster nanoparticle The method and its application in clinical detection, diagnosis and treatment. [Prior Art] Magnetic resonance imaging is a non-invasive and non-radiative free contrast technique. It has good imaging contrast, high spatial resolution, and ability tomography in soft tissue. It is a clinical diagnosis in recent years. A very important imaging tool. The developers commonly used in clinical practice today are mainly terpenoid developers, and cockroaches are toxic to cells. If the dosage is improperly used or the dosage form is poorly designed, it is likely to cause serious side effects and affect human health. Moreover, the use of guanidine-based developers sometimes causes "pseudo-signal" or the disadvantage of diluting the concentration due to the secretion of the body, causing the signal to disappear. Therefore, the development and application of other developers have become an important issue in magnetic resonance imaging technology. One of the major developmental magnetic resonance imaging agents currently is superparamagnetic iron oxide (SPIO) nanoparticles. In clinical experiments, it can be applied to molecular imaging, gene labeling, non-invasive cell labeling or tumor detection, etc., and its application range is quite extensive. The known process of superparamagnetic iron oxide nanoparticles produces uniform distribution of nanoparticles in the organic phase, and in the aqueous phase, the cluster type is predominant. Although a protective layer can be formed on the surface of the particle by adding a polymer or a surfactant to obtain a relatively dispersed nanoparticle in an aqueous solution, since the surface of the particle 201244740 is protected by a polymer or a surfactant molecule, it is not easy to be Further application. Furthermore, unmodified or encapsulated; scattered, will limit its use in the organism. However, most of the m = is a soluble substance, and it is very important to apply such a nanoparticle to 3. The technology of the ship is not the particles [invention] The cluster-like ί provides a mixed solution or aqueous solution of a kind of water-soluble super-stable nano-particles and water-soluble superparamagnetic iron oxide, an emulsified solution, in which the emulsified wave U Loose' makes it a super-paramagnetic iron oxide too many small oil droplets, these small oil drops in vitamin E. The dancers are dispersed and coated with water so that the small oil droplets::, the oil phase solvent of the emulsified solution is heated to remove each of them. One embodiment of the present invention, wherein the oil phase solvent is an aliphatic acid solution, wherein the aliphatic solvent comprises bismuth, dioxane or n-hexane. According to one embodiment of the present invention, wherein the lipid steroid solvent comprises Zheng 201244740 hexane. According to an embodiment of the invention, the aqueous solution comprises a solution of triethylamine, an aqueous solution of doxorubicin or any combination of the above. According to an embodiment of the invention, the volume ratio of n-hexane to deionized water or aqueous solution is from 1:50 to 1:1000. According to an embodiment of the invention, the volume ratio of n-hexane to deionized water or aqueous solution is 1:500. In accordance with an embodiment of the invention, the ultrasonic is shocked for 5 minutes to 15 minutes. According to an embodiment of the invention, the temperature at which the above mixed solution is heated is from 65 ° C to 105 ° C. According to an embodiment of the invention, the above mixed solution is heated at 75 ° C for 1 hour. According to an embodiment of the present invention, the polydispersity index of the water-soluble cluster nanoparticles described above is from 0.2 to 0.5. Still another aspect of the present invention is to provide a water-soluble superparamagnetic clustered nanoparticle which is obtained by the above method. Still another aspect of the present invention provides a magnetic resonance contrast developer comprising the water-soluble superparamagnetic cluster nanoparticle produced by the above method and water. Another aspect of the present invention provides a pharmaceutical composition comprising the water-soluble superparamagnetic cluster nanoparticles prepared by any of the above production methods and examples, and an active material, and the active material is Coated in water-soluble superparamagnetic cluster nanoparticles. According to an embodiment of the invention, the active substance is Agrobacterium 201244740. The solvent is used to form a superparamagnetic iron oxide 2::na:particle' which is w-coated in a water-soluble vitamin column and is dispersed in the water-soluble super-smoky nucleus cluster particle. When the best granules are applied to the magnetic resonance imaging developer clusters, the nanoparticles can also be used as =::=====hydrophobic _) coated in the water Nilaiwei cluster nano-Lt In the present invention, the superparamagnetic cerium oxide nanoparticles and the active material (for example, a hydrophobic drug) are coated with a water-soluble vitamin E, so that the super-magnetic nano particles are soluble and safe. It is a water-soluble superparamagnetic clustered nanoparticle, which can be stably and uniformly dissolved in a physiological buffer solution or other aqueous solution, and can be used in clinical detection, diagnosis and treatment. In nanoparticles, changing the surface chemistry can affect the properties of nanoparticles, such as changing physical and chemical properties, including size, solubility, dispersion state, and magnetization values. Therefore, by changing the surface chemistry, the properties of the magnetic nanoparticles in the biological system can be changed, including cell recognition, biodistribution, and immune response to reduce the potential nanotoxicity of the nanoparticles.
四 α -生育酚聚乙二醇琥珀酸（D- a -tocopheryl polyethylene glycol 1000 succinate » TPGS 1000 ； Vitamin E 201244740 TPGS)為水溶性維生素E ’具有親水端和親脂端。其具有表 面活性劑之特性，因此可作為增溶劑、乳化劑和脂溶性藥 物載體之給藥製劑。除了表面活性劑的特性以外，此水溶 性維生素E可有效促進生物體對藥物的吸收和利用並降低 生物體的抗藥性。 超順磁性氧化鐵奈米粒子可經由購買或自行合成取 得’製備過程可參照Sun和Zeng (2002)所發表之方法。將乙 醯丙酮鐵(Fe(acac)3)和 1，2-十六烷二醇（1,2-hexadecanediol) 以莫耳比1.1 〇加到20 mL的本基越(phenyl ether)中均勻混 合，加熱至約60°C，使混合物完全溶於苯基醚後，再加入2 mL油相界面活性劑油酸（〇ldc add)和2紅油胺 (〇kylamine)，通入氮氣並緩慢升溫至250。(：，持續反應3小 時後，可合成直徑約6 rnn的超順磁性氧化鐵奈米粒子。 水溶性超順磁性群集奈米粒子之製造方法 性氧===為依照本發明一實施例之水溶性超順磁 固粒子的製造方法之流程圖。步驟110為將 米粒子和水溶性維生素E混入油相 鐵奈米教+Γίί ㈣。在步驟ug中’超順磁性氧化 均i混合，Γ加’先在室溫下 其具有油⑽姊==或水錢，形成^合溶液， 液。其中乳波震盤混合溶液，使其形成乳化溶D-a-tocopheryl polyethylene glycol 1000 succinate (TPGS 1000; Vitamin E 201244740 TPGS) is a water-soluble vitamin E' having a hydrophilic end and a lipophilic end. It has the characteristics of a surfactant, and thus can be used as a preparation preparation of a solubilizing agent, an emulsifier, and a fat-soluble drug carrier. In addition to the characteristics of the surfactant, the water-soluble vitamin E can effectively promote the absorption and utilization of the drug by the organism and reduce the resistance of the organism. Superparamagnetic iron oxide nanoparticles can be obtained by purchase or by self-synthesis. The preparation process can be referred to the method published by Sun and Zeng (2002). Ethylacetone iron (Fe(acac)3) and 1,2-hexadecanediol (1,2-hexadecanediol) were uniformly mixed in 20 mL of phenyl ether with a molar ratio of 1.1 〇. Heat to about 60 ° C, after the mixture is completely dissolved in phenyl ether, add 2 mL of the oil phase surfactant oleic acid (〇ldc add) and 2 erythroylamine (〇kylamine), pass nitrogen and slowly heat up To 250. (:, after continuous reaction for 3 hours, superparamagnetic iron oxide nanoparticles having a diameter of about 6 rnn can be synthesized. Method for producing water-soluble superparamagnetic clustered nanoparticles. Oxygen === according to an embodiment of the present invention A flow chart of a method for producing water-soluble superparamagnetic solid particles. Step 110 is to mix rice particles and water-soluble vitamin E into the oil phase of T. sinensis + Γίί (4). In step ug, 'superparamagnetic oxidation is mixed, Γ Add 'first at room temperature, it has oil (10) 姊 == or water money to form a solution, liquid. The milk wave shakes the solution to make it form an emulsified solution.
性氧化鐵奈米粒子二二:小油滴’小油滴内之超順磁 中。 ’、刀政排列且包覆於水溶性維生素E 8 201244740 步驟130為加熱乳化溶液，使乳化溶液 * 全揮發。此時超順磁性氧化鐵奈米粒 _目:劑完 成群集奸(d_i·)。 象而形 =14〇為使小油滴形成水溶性超順磁 子。在步驟130產生群聚現象後，小油滴即形成=:t粒 性超順磁性群集奈米粒子’其,每一水溶性=群= 奈米粒子之超順磁絲化鐵奈米粒子係呈 群集 於該水溶性維生素E中。 彳且包覆 依照本發明之-個或多個實施例，油相 溶劑，而脂肪族溶劑包含乙酸乙酯、四氫呋喃、馬知肪族 或正己烷。 、二氣曱烷 請參照表一，為水溶性超順磁性氧化鐵群 ” 之製備條件，其中油相溶劑是正己烧。多分散、米板子 (polydispersity index ; PDI)為利用雷射粒徑分析^生指數 laser scattering ; DLS)量測溶液中奈米粒子半徑，=yna功ic 換算所得之數值。若所有奈米粒子粒徑愈—致，^比對與 性指數愈接近0，分散性較小，則製成的奈米粒子>、多分散 為均勻；若奈米粒徑不一致，其PDI就越貼近於1之杈後較 較大’則製成的奈米粒子之粒徑較不均勻。 分散性 表一 、水溶性超順磁性氧化鐵群集奈米粒子之製備條件 正己烧加熱板溫度沸騰所需時間溶液溫度^ =(秒） (°〇 (nmf 多分 1 2 3 4 2 2 2 1 550 400 240 550 55 121 285 55 102.5 97.1 88.1 102.5 22) °·484 °·416 °·396 0.489Iron oxide nanoparticles 22: small oil droplets in the super-paramagnetic in small oil droplets. ‘, knife arrangement and coated with water-soluble vitamin E 8 201244740 Step 130 is to heat the emulsified solution, so that the emulsified solution * is completely volatilized. At this time, the superparamagnetic iron oxide nanoparticle _ mesh: the agent completes the cluster rape (d_i·). Elephant shape = 14 〇 is to make small oil droplets form water-soluble superparamagnetic. After the clustering phenomenon occurs in step 130, the small oil droplets form =: t-granular superparamagnetic cluster nanoparticles, which are each of the water-soluble = group = nanoparticle superparamagnetic silk fibroin nanoparticle system Clustered in the water-soluble vitamin E. And coating the oil phase solvent according to one or more embodiments of the present invention, and the aliphatic solvent comprises ethyl acetate, tetrahydrofuran, mazinol or n-hexane. For the dioxane, please refer to Table 1 for the preparation conditions of the water-soluble superparamagnetic iron oxide group, in which the oil phase solvent is positively burned. The polydispersity index (PDI) is analyzed by laser particle size. ^sheng index laser scattering; DLS) Measure the radius of the nanoparticle in the solution, = yna ic converted to the value. If all the nanoparticles have a larger particle size, the ratio of ^ is closer to 0, the dispersion is better. Small, the prepared nanoparticles are >, and the polydispersity is uniform; if the nanometer particle size is inconsistent, the PDI is closer to 1 and then larger, then the particle size of the prepared nanoparticle is less uniform. Dispersibility Table 1. Preparation conditions of water-soluble superparamagnetic iron oxide cluster nanoparticles. Time required for boiling of hot plate temperature. Solution temperature ^ = (seconds) (°〇(nmf multi-point 1 2 3 4 2 2 2 1) 550 400 240 550 55 121 285 55 102.5 97.1 88.1 102.5 22) °·484 °·416 °·396 0.489
231.9 188.6 192.6 248.0 9 201244740 121 97.1 285 88.1 55 102.5 121 97.1 285 88.1 121 97.1 285 88.1 375 80.5 1011 67.2 285 88.1 285 88.1 289.0 0.612 166.5 0.228 167.4 0.486 152.8 0.281 166.1 0.322 128.0 0.413 188.8 0.403 103.3 0.323 117.1 0.287 164.3 0.552 122.5 0.422 5 1 400 6 1 240 7 0.4 550 8 0.4 400 9 0.4 240 10 0.2 400 11 0.2 240 12 0.2 200 13 0.2 160 14 0.1 240 15 0.05 240 、根據本發明之一實施例，製備過程是先將5呵的超川員 磁性氧化鐵奈米粒子和20 mg的水溶性維生素£加入〇2 之正己烧内，在室溫下均勻混合，再加入1〇紅之去離子 水，以超音波震盪10分鐘形成乳化溶液，其中乳化溶液包 括複數個小油滴’這些小油滴内之超順磁性氧化鐵奈来^ 子係呈分散排列且包覆於水溶性維生素。接著，於 加溫1小時’使混合溶液中的正己烧完全揮發。製備完成後 冷卻至室溫，以冷;東乾燥將水分絲。最後將冷綠燥後 的產物回溶於水中或生理緩衝溶液。 、請參照附件1，為水溶性超順磁性群集奈米粒子之穿透 式電子顯微鏡(Transmission electron microscopy ; TEM)影 像圖。圖中顯示經油相溶劑揮發後，水溶性超順磁性群集 奈米粒子之超順磁性氧化鐵奈米粒子係呈規則排列且包覆 於該水溶性維生素E中，而形成規則排列之群集（—ter)奈 201244740 磁共振造影顯影劑 奈米和溶劑揮發的方法將超順磁性氧化鐵 *互溶的系使水相和油相兩 液中，再將油相的_揮發使原本㈣於水溶 化鐵奈米粒子形成群集粒子政：超順磁性氧 鐵群集奈米粒子的顯影效果較分散排化 而得到較好的顯影效果。 π之^切子更佳’ 影像為性群集奈米粒子之磁振 ==超順磁性群集奈米粒子’鐵濃度自=Γ:依 奈米粒子，鐵=0)為低，度之水溶性超順磁性群集 鐵濃度的降低^弱。Mgm依序減低，而顯影效果也依 請讀n水雜_磁 f細胞株Mc™抓之磁振影像圖贺 醫師所提供’其為具多重抗藥性的 礼3胞株。細胞在處理水溶性超順磁性氧化鐵 粒子後，於各時間點收取細胞， Μ /、/、未 造影(請參照附件3)，其中對照組為 ?小時的細胞。各時間點樣本影像二 數:愈小，代表顏色越黑，其顯影效果愈佳： rti: ? 長，細胞的顯影效果愈佳，此表示癌231.9 188.6 192.6 248.0 9 201244740 121 97.1 285 88.1 55 102.5 121 97.1 285 88.1 121 97.1 285 88.1 375 80.5 1011 67.2 285 88.1 285 88.1 289.0 0.612 166.5 0.228 167.4 0.486 152.8 0.281 166.1 0.322 128.0 0.413 188.8 0.403 103.3 0.323 117.1 0.287 164.3 0.552 122.5 0.422 5 1 400 6 1 240 7 0.4 550 8 0.4 400 9 0.4 240 10 0.2 400 11 0.2 240 12 0.2 200 13 0.2 160 14 0.1 240 15 0.05 240 According to an embodiment of the invention, the preparation process is 5 的 first Superchuan magnetic iron oxide nanoparticles and 20 mg of water-soluble vitamins are added to the 己2 of the hexahydrate, uniformly mixed at room temperature, then added with 1 blush deionized water, vortexed by ultrasonic for 10 minutes to form an emulsion. A solution in which the emulsified solution comprises a plurality of small oil droplets. The superparamagnetic iron oxides in these small oil droplets are dispersed and coated with a water-soluble vitamin. Then, the autoclave in the mixed solution was completely volatilized by heating for 1 hour. After the preparation is completed, it is cooled to room temperature to be cold; the east is dried to moisture. Finally, the product after cold green drying is dissolved in water or a physiological buffer solution. Please refer to Annex 1 for a transmission electron microscopy (TEM) image of water-soluble superparamagnetic cluster nanoparticles. The figure shows that after the oil phase solvent is volatilized, the superparamagnetic iron oxide nanoparticles of the water-soluble superparamagnetic cluster nanoparticles are regularly arranged and coated in the water-soluble vitamin E to form a regularly arranged cluster ( —ter)奈 201244740 Magnetic Resonance Imaging Developer Nanoparticle and Solvent Volatilization Method The superparamagnetic iron oxide* miscible system is used to make the aqueous phase and the oil phase two liquids, and then the oil phase is volatilized to make the original (4) water soluble. Iron nanoparticle formation cluster particle politics: The development effect of superparamagnetic oxyiron cluster nanoparticle is better than that of dispersion and discharge. π^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ The decrease in iron concentration in the paramagnetic cluster is weak. The Mgm is sequentially reduced, and the development effect is also dependent on the magnetic resonance image of the Mc-cell strain, which is provided by Dr. Physician, which is a multi-drug resistant cultivar. After treating the water-soluble superparamagnetic iron oxide particles, the cells were harvested at various time points, Μ /, /, and no contrast (see Annex 3), in which the control group was ? hour cells. The number of sample images at each time point: the smaller, the darker the color, the better the development effect: rti: ? Long, the better the development of the cells, which means cancer
入水溶性超姆氧化鐵群集奈米粒子，而產2 顯影效果。 H 201244740 表二、水溶性超順磁性氧化鐵群集奈米粒子於MCF_7/ADR之顯影 效果 對照組 1小時 4小時 6小時 12小時 樣本 103.19 90.34 89.61 88.81 87.75 樣本/背景 0.924 0.865 0.882 0.879 0.832 相對倍數變化 1.00 0.94 0.96 0.95 0.90 醫藥組成物 運用本發明亦可將活性物質（例如疏水性藥物，如阿徽 素（doxorubicin))包覆在水溶性維生素e中，應用於抗癌治 療。阿黴素為一種抗腫瘤抗生素’可抑制去氧核醣核酸和 核糖核酸的合成，對多種腫瘤均有殺滅作用，可用以治療 多種癌症。 製備過程是將5 mg的超順磁性氧化鐵奈米粒子和加 mg的水溶性維生素E加入0.2 mL之正己烧内，在室溫下土勺 勻混合’再加入含有10//L三乙胺（triethylamine)溶液和 0.5mg阿黴素混合之水溶液（10 ml) ’以超音波震盪1〇分鐘形 成乳化態。接著’於75°C加溫1小時，使混合溶液中的正己 院完全揮發。製備完成後冷卻至室溫，以冷康乾燥將水分 去除。將冷凍乾燥後的產物回溶於水中或生理緩衝溶液 (phosphate buffered saline ; PBS)。 請參照第2圖，為水溶性超順磁性群集奈米粒子處理細 胞株MCF-7/ADR之細胞存活率圖。此項試驗是利用Μττ (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)分析測定癌細胞在處理水溶性超順磁性氧化鐵群 12 201244740 集奈米粒子後的存活率。由第2圖 順磁性氧化鐵群集奈錄之結果顯示，以水溶性超 鐵含量越高，賴細胞存活^癌細胞’當奈米粒子中的 由上述本發明實施方式可A越低。 順磁性群集奈米粒子，其優點°，應用本發明的水溶性超 覆超順磁性氧化鐵奈米粒子;利用水溶性維生素E包 溶劑揮發，使原本排列分散^其可水溶性’並透過油相 成規則排狀群餘子，而㈣磁性氧化鐵奈米粒子形 氧化鐵奈米粒子。除此之外1顯影效果較佳的超順磁性 子亦可包覆疏水性藥物，而 ☆性超順磁性群集奈米粒 系統的潛力。 "、發展成為多功能給藥載體 …太^本發明已以實施方式揭露如上，然其並非用以 :本’在本發明所屬技術領域中任何具有通常知： 與^飾’因此本發明之保護範圍當視後附之中 2 所界定者為準。 』軏圍 【圖式簡單說明】 為讓本發明之上述和其他目的、特徵、優點與實施例 能更明顯易懂’所附圖式之說明如下： 第1圖為依照本發明一實施例之製備水溶性超順磁性 氧化鐵群集奈米粒子之方法流程圖。 第2圖為水溶性超順磁性氧化鐵群集奈米粒子處理細 胞株MCF-7/ADR之細胞存活率圖。 附件1為水溶性超順磁性群集奈米粒子之穿透式電子 201244740 顯微鏡影像圖。 附件2為水溶性超順磁性群集奈米粒子之磁振影像 圖。 附件3為水溶性超順磁性氧化鐵群集奈米粒子處理細 胞株MCF-7/ADR之磁振影像圖。 【主要元件符號說明】 120 :步驟 140 :步驟 110 :步驟 130 :步驟 14Into the water-soluble super oxidized iron cluster nano particles, and produce 2 development effects. H 201244740 Table 2. Development effect of water-soluble superparamagnetic iron oxide cluster nanoparticles on MCF_7/ADR control group 1 hour 4 hours 6 hours 12 hours sample 103.19 90.34 89.61 88.81 87.75 sample / background 0.924 0.865 0.882 0.879 0.832 relative fold change 1.00 0.94 0.96 0.95 0.90 Pharmaceutical Compositions The present invention can also be used for anticancer treatment by coating an active substance (for example, a hydrophobic drug such as doxorubicin) in water-soluble vitamin E. Doxorubicin is an anti-tumor antibiotic that inhibits the synthesis of DNA and ribonucleic acid and has a killing effect on a variety of tumors, and can be used to treat a variety of cancers. The preparation process is to add 5 mg of superparamagnetic iron oxide nanoparticles and mg of water-soluble vitamin E to 0.2 mL of hexose, mix thoroughly at room temperature, and then add 10//L triethylamine. An aqueous solution (10 ml) of a mixture of (triethylamine) solution and 0.5 mg of doxorubicin was emulsified by ultrasonic waves for 1 minute to form an emulsified state. Then, the temperature was raised at 75 ° C for 1 hour to completely evaporate the positive chamber in the mixed solution. After the preparation was completed, it was cooled to room temperature, and the water was removed by cold drying. The lyophilized product was dissolved back in water or phosphate buffered saline (PBS). Please refer to Fig. 2 for the cell viability map of the water-soluble superparamagnetic cluster nanoparticles treated cell line MCF-7/ADR. This test uses Μττ (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to determine the cancer cells after treatment of water-soluble superparamagnetic iron oxide group 12 201244740 nanoparticles. Survival rate. From Fig. 2, the results of the paramagnetic iron oxide cluster show that the higher the water-soluble super-iron content, the lower the survival rate of the lysed cells in the cancer cells as in the above-mentioned embodiments of the present invention. Paramagnetic clustered nanoparticle, the advantage thereof, the water-soluble super-superparamagnetic iron oxide nanoparticle of the invention is applied; the water-soluble vitamin E is used to volatilize the solvent to make the original arrangement disperse|the water-soluble' and oil-permeable The phase is formed into a regular row of clusters, while (iv) magnetic iron oxide nanoparticle-shaped iron oxide nanoparticles. In addition to this, a superparamagnetic agent with better development effect can also coat hydrophobic drugs, and the potential of the ☆ super-paramagnetic cluster nanoparticle system. ", has developed into a multi-functional drug delivery carrier. The present invention has been disclosed in the above embodiments, but it is not intended to be used in the technical field of the present invention. The scope of protection shall be subject to the definition in 2 of the attached. BRIEF DESCRIPTION OF THE DRAWINGS The above and other objects, features, advantages and embodiments of the present invention will become more <RTIgt; A flow chart of a method for preparing water-soluble superparamagnetic iron oxide cluster nanoparticles. Fig. 2 is a graph showing the cell survival rate of the water-soluble superparamagnetic iron oxide cluster nanoparticle-treated cell strain MCF-7/ADR. Annex 1 is a penetrating electron of water-soluble superparamagnetic cluster nanoparticles. 201244740 Microscope image. Annex 2 is a magnetic image of water-soluble superparamagnetic cluster nanoparticles. Annex 3 is a magnetic resonance image of the water-soluble superparamagnetic iron oxide cluster nanoparticle-treated cell strain MCF-7/ADR. [Main component symbol description] 120: Step 140: Step 110: Step 130: Step 14