TWI473763B - 樹枝狀高分子與石墨烯氧化物的螢光混成物 - Google Patents
樹枝狀高分子與石墨烯氧化物的螢光混成物 Download PDFInfo
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims description 28
- 229910021389 graphene Inorganic materials 0.000 title claims description 28
- 239000000412 dendrimer Substances 0.000 title claims description 18
- 229920000736 dendritic polymer Polymers 0.000 title claims description 18
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 173
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 87
- 235000019152 folic acid Nutrition 0.000 claims description 87
- 239000011724 folic acid Substances 0.000 claims description 87
- 229960000304 folic acid Drugs 0.000 claims description 87
- 239000000463 material Substances 0.000 claims description 59
- 239000000203 mixture Substances 0.000 claims description 58
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 18
- -1 poly(amidinoamine) Polymers 0.000 claims description 17
- 230000005284 excitation Effects 0.000 claims description 15
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 14
- 238000010521 absorption reaction Methods 0.000 claims description 14
- 238000002428 photodynamic therapy Methods 0.000 claims description 12
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical class CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 7
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- VHYFNPMBLIVWCW-UHFFFAOYSA-O n,n-dimethylpyridin-1-ium-4-amine Chemical compound CN(C)C1=CC=[NH+]C=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-O 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 239000003550 marker Substances 0.000 claims description 3
- 150000001412 amines Chemical class 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 229920000877 Melamine resin Polymers 0.000 claims 2
- JDSHMPZPIAZGSV-UHFFFAOYSA-N melamine Chemical group NC1=NC(N)=NC(N)=N1 JDSHMPZPIAZGSV-UHFFFAOYSA-N 0.000 claims 2
- 239000002253 acid Substances 0.000 claims 1
- 125000003277 amino group Chemical group 0.000 claims 1
- 238000005286 illumination Methods 0.000 claims 1
- 239000013067 intermediate product Substances 0.000 claims 1
- NAYYNDKKHOIIOD-UHFFFAOYSA-N phthalamide Chemical compound NC(=O)C1=CC=CC=C1C(N)=O NAYYNDKKHOIIOD-UHFFFAOYSA-N 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 21
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 18
- 229920000962 poly(amidoamine) Polymers 0.000 description 18
- SENLDUJVTGGYIH-UHFFFAOYSA-N n-(2-aminoethyl)-3-[[3-(2-aminoethylamino)-3-oxopropyl]-[2-[bis[3-(2-aminoethylamino)-3-oxopropyl]amino]ethyl]amino]propanamide Chemical compound NCCNC(=O)CCN(CCC(=O)NCCN)CCN(CCC(=O)NCCN)CCC(=O)NCCN SENLDUJVTGGYIH-UHFFFAOYSA-N 0.000 description 13
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- FTZIQBGFCYJWKA-UHFFFAOYSA-N 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Chemical compound S1C(C)=C(C)N=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 FTZIQBGFCYJWKA-UHFFFAOYSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- PEECTLLHENGOKU-UHFFFAOYSA-N n,n-dimethylpyridin-4-amine Chemical compound CN(C)C1=CC=NC=C1.CN(C)C1=CC=NC=C1 PEECTLLHENGOKU-UHFFFAOYSA-N 0.000 description 1
- KOOMFXGDLMRWSN-UHFFFAOYSA-N n-phenylnitrous amide Chemical compound O=NNC1=CC=CC=C1 KOOMFXGDLMRWSN-UHFFFAOYSA-N 0.000 description 1
- 125000000018 nitroso group Chemical group N(=O)* 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G73/00—Macromolecular compounds obtained by reactions forming a linkage containing nitrogen with or without oxygen or carbon in the main chain of the macromolecule, not provided for in groups C08G12/00 - C08G71/00
- C08G73/02—Polyamines
- C08G73/028—Polyamidoamines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/002—Dendritic macromolecules
- C08G83/003—Dendrimers
- C08G83/004—After treatment of dendrimers
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/02—Elements
- C08K3/04—Carbon
- C08K3/042—Graphene or derivatives, e.g. graphene oxides
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- C08K5/00—Use of organic ingredients
- C08K5/16—Nitrogen-containing compounds
- C08K5/34—Heterocyclic compounds having nitrogen in the ring
- C08K5/3442—Heterocyclic compounds having nitrogen in the ring having two nitrogen atoms in the ring
- C08K5/3462—Six-membered rings
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/02—Elements
- C08K3/04—Carbon
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
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Description
本發明是有關於一種混成材料,且特別是有關於一種石墨烯氧化物與樹枝狀高分子的螢光混成材料。
石墨烯氧化物(graphene oxide,GO)是一種氧化的石墨烯主體材料。氧化的石墨烯主體材料是由經含氧基團化學官能化的類石墨烯片材(graphene-like sheet)所組成,所述的含氧基團諸如是羥基、羧酸以及環氧化合物之基團。已有報導指出石墨烯氧化物的官能化會導致其電性與光學性的改變。如本發明所提供的,石墨烯氧化物的混成材料可作為生物與醫學應用上有前途的新材料,包含細胞顯影、藥物傳遞以及光動力療法之應用。
樹枝狀高分子典型是環繞著核心對稱的高度分支化的分子,且通常採取球形三維形貌。聚(醯胺基胺)(poly(amidoamine),PAMAM)樹枝狀高分子一般具有多重官能化的球形表面、高度的分子均勻性、明確定義的分子量以及具體的尺寸與形狀。PAMAM樹枝狀高分子的製造流程包括一系列由中央起始核心展開的重覆步驟。每一套重覆步驟造成一內層(意即,一”代”),所述的內層是由加至核心的重複單元所組成,使得聚合物具有較大的分子直徑、兩倍數量的活性表面部位以及前一代聚合物的接近兩倍分子量。樹枝狀高分子已在化學以及生物領域中引起
廣泛的興趣,特別是於藥物傳遞、基因療法以及化學療法之應用。
PAMAM樹枝狀高分子也具有強的螢光放射而使其成為潛力顯影劑。螢光現象主要由PAMAM樹枝狀高分子之三級胺基團的氧化作用所貢獻。由於此螢光特性,PAMAM樹枝狀高分子的胞噬行為可直接地藉由螢光顯微技術伴隨流式細胞技術來分析,而不需額外的螢光標記。
然而,對於生物或生醫應用而言,樹枝狀高分子的生物相容性,包含細胞毒性、生物可降解能力以及活體內新陳代謝,必須與樹枝狀高分子的水溶性以及官能度取得平衡。
本發明是有關於一種新的雙功能混成材料。上述混成材料可同時作用為螢光標記或探針以及作用為攻擊者。
本發明提出一種混成材料。上述混成材料包含至少一端羥基***(G-4)聚(醯胺基胺)(PAMAM)樹枝狀高分子(DEN-OH)以及葉酸(FA)化學鍵結於石墨烯氧化物(GO)上。製作混成材料是藉由混合DEN-OH以及GO並於室溫中攪拌,添加N,N’-二環己基碳二亞胺(DCC)以及4-二甲基氨基吡碇(DAMP)至DEN-OH與GO的混合物並於室溫中攪拌以進行酯化作用以形成中間產物GO/DEN-OH,GO/DEN-OH是指DEN-OH鍵結於GO上,添加N-羥基丁二醯亞胺(NHS)以及1-(3-二甲氨基丙基)-3-乙基碳二亞胺
鹽酸鹽(EDC)至中間產物GO/DEN-OH之水性懸浮液中,接著加入葉酸至NHS、EDC與中間產物GO/DEN-OH之水性懸浮液的混合物中,且於室溫中攪拌以獲得混成材料。
本發明提供奈米混成材料,該奈米混成材料顯現出雙光子螢光放射並且能夠在接受照射後產生活性含氧物(reactive oxygen species)。
為讓本發明之上述特徵和優點能更明顯易懂,下文特舉實施例,並配合所附圖式作詳細說明如下。
本發明提供包含PAMAM樹枝狀高分子鍵結至石墨烯氧化物(GO)的新混成材料。在本發明中,端羥基***(***,G-4)PAMAM樹枝狀高分子(DEN-OH)與葉酸(folic acid,FA)化學鍵結至石墨烯氧化物(GO)上以形成混成材料。
這樣的混成材料可透過經鍵結之葉酸特定地靶向具葉酸受體的細胞。再者,混成材料於雷射照射下產生活性含氧物的能力使其成為光動力療法的潛力製劑。
為了降低細胞毒性,本發明採取端羥基PAMAM樹枝狀高分子製備混成材料,而非有毒的端胺基(amine-terminated)樹枝狀高分子。端羥基PAMAM樹枝狀高分子是藉由GO的-COOH基團與端羥基PAMAM樹枝狀高分子的羥基基團之間的共價鍵結而連接至石墨烯氧化物上。葉酸經由GO的-COOH基團與葉酸的NH2
基團反應而
獨立連接至石墨烯氧化物。對本發明所製備之混成材料而言,石墨烯氧化物為承載平台。石墨烯氧化物所載之樹枝狀高分子以作為石墨烯氧化物在水中的懸浮劑,而石墨烯氧化物所載之葉酸以攻擊目標癌細胞。本發明所製備的混成材料為水可分散性(water dispersible)。作為藥物傳遞系統時,藥物可載於石墨烯氧化物或載於經鍵結的樹枝狀高分子,且可同時運送多種藥物至目標部位。
由於本發明所製備的混成材料中的經氧摻雜PAMAM樹枝狀高分子具有螢光,因此本混成材料不需額外的染料或是螢光標誌。這樣是有益的,因為額外的染料通常會增加毒性。因為PAMAM樹枝狀高分子的關係,混成材料顯現出雙光子吸收性,且混成材料在雷射照射下產生能夠攻擊細胞的活性含氧物。
GO是具氧化基團的氧化石墨烯片材。氧化基團像是羥基、羧酸以及環氧化物之基團,且GO的氧化基團的含量或數量取決於氧化程度。***端羥基PAMAM樹枝狀高分子(G-4 DEN-OH)之分子量為14,277 Da,其分子式為C622
H1184
N186
O188
,具有64個末端基團。
如圖1所描述,DEN-OH以及葉酸藉由使用縮合劑分別進行酯化作用以及醯胺化作用以連接至GO。
使DEN-OH甲醇溶液(10 wt%,1 cm3
)的溶劑蒸發後,以二甲基甲醯胺(DMF)(50 cm3
)溶解殘留物。將GO水性分散液(4 cm3
)離心後,移除上清液並將離心分離物分散於DMF(4 cm3
)中。於室溫中攪拌以混合樹枝狀高分子的DMF溶液(0.2 wt%,6 cm3
)與GO的DMF溶液(10 wt%,4 cm3
)。接著,加入N,N’-二環己基碳二亞胺(N,N’-dicyclohexylcarbodiimide,DCC)(範圍介於3.7 mg至18.5 mg之間)以及4-二甲基氨基吡碇(4-dimethylaminopyridine,DMAP)(約DCC的十分之一)後,將混合物於室溫中強力地攪拌三天以進行酯化作用。將上述分散液離心,並將離心分離物(GO/DEN-OH)以DMF潤洗數次後,再將離心分離物(GO/DEN-OH)分散於水中。
已合成之GO/DEN-OH/FA混成物可具有不同濃度的DEN-OH。以DEN-OH濃度為18 μM的GO/DEN-OH/FA混成物而言,GO:DEN-OH:FA的相對比例為0.55 mg:2.5 mg:0.45 mg。以DEN-OH濃度為90 μM的GO/DEN-OH/FA混成物而言,GO:DEN-OH:FA的相對比例為0.55 mg:12.8 mg:0.45 mg。意即,混成材料包括50-256重量份的端羥基聚(醯胺基胺)(PAMAM)樹枝狀高分子(DEN-OH)、9重量份的葉酸(FA)以及11重量份的石墨烯氧化物(GO),其中DEN-OH以及FA是化學鍵結至GO上以形成混成材料。
將N-羥基丁二醯亞胺(NHS)(182.5 mg,1.6 mmol)以及1-(3-二甲氨基丙基)-3-乙基碳二亞胺鹽酸鹽(EDC)(125 mg,
0.65 mmol)加入GO/DEN-OH的水性懸浮液(2 cm3
)中,並將此混合物超音波震盪2小時。然後,將葉酸(0.5 wt%,8 cm3
)的10 wt%NaHCO3
水性溶液(pH 8)加入混合物中並於室溫中攪拌過夜。產物(GO/DEN-OH/FA)純化是以NaHCO3
溶液(pH 8)進行透析48小時後接著以水透析24小時來實現。GO上鍵結FA也藉由相同的步驟來實現純化。
圖2繪示具有不同DEN-OH濃度的GO/DEN-OH/FA之混成物的紫外光可見光吸收光譜。在FA被載於GO/DEN-OH上後,每一個GO/DEN-OH/FA混成物都於約280 nm之處出現吸收帶。
在DEN-OH與葉酸固定於GO之後,該經DEN-OH與葉酸固定的GO的發光性質被研究出具有不同的激發波長。圖3A至圖3E所表示的是,在不同激發波長下,不同DEN-OH濃度(DEN-OH為18 μM以及90 μM)的GO/DEN-OH/FA混成物所顯示的放射光譜。GO/DEN-OH/FA混成物於230 nm以及280 nm的激發波長下於355 nm以及450 nm顯現出強度相當的放射能帶。然而,GO/DEN-OH/FA混成物於360 nm、390 nm以及780 nm的激發波長下只顯現出位於450 nm的一個放射能帶。
螢光放射是一種能量釋放現象,其是藉由分子吸收光
線或其他電磁波,然後發生波長大於吸收光線的單光子放射。然而,當所吸收的電磁波夠強烈時,可能會導致一個分子吸收兩個光子。此雙光子吸收能夠導致波長短於所吸收電磁波長的放射。在雙光子激發的方式上,典型是於近紅外光範圍內且於足夠強度的雷射光下,螢光團同時吸收能量較低的兩個光子而激發。由於雙光子訊號微弱,因此雙光子吸收最常透過誘發螢光來觀察。倘若兩個光子的能量總合大於分子的基態與激發態之間的能量間隙,則可發生非線性過程。一般而言,雙光子激發的波長約為單光子激發波長的超過兩倍。
GO/DEN-OH/FA混成材料的雙光子吸收性質是使用雙光子激發顯微技術測量在720-990 nm的激發波長來觀察發亮的螢光。圖4表示GO/DEN-OH/FA混成物(DEN-OH為90 μM)的雙光子激發螢光放射影像,且圖5顯示使用影像軟體分析以雙光子激發的可見光螢光GO/DEN-OH/FA的數量。觀察到可見光螢光GO/DEN-OH/FA的數量對應於不同激發波長而改變,且GO/DEN-OH/FA混成材料於760-920 nm處出現螢光放射。
由於GO/DEN-OH/FA混成物於近紅外光(NIR)區域的強烈螢光放射,GO/DEN-OH/FA混成物可於生醫應用上作為顯影劑或是螢光標記。GO/DEN-OH/FA混成物的雙光子吸收現象可被開發並進一步應用於雙光子螢光顯影及/或雙光子光動力癌症療法中。
GO/DEN-OH/FA混成材料的雙光子吸收性是由使用
螢光光譜儀於390 nm以及780 nm激發波長都有450 nm的放射峰而確認。GO/DEN-OH/FA混成物放射出短於吸收光之波長的螢光,因此確認了GO/DEN-OH/FA混成物具有強烈的雙光子吸收性。
光動力療法是用於包含癌症的許多疾病的非侵入式療程。一般而言,光動力療法涉及向光敏製劑或光感物質照射恰當波長的可見光或近紅外光。由於GO/DEN-OH/FA混成物擁有雙光子吸收性質且為水可分散性的,這些混成物適於被使用作為光動力療法的光敏製劑或光感物質。於光動力療法的活體內評估中,會檢查GO/DEN-OH/FA混成物與海拉細胞(HeLa cells)的交互作用,其中混成物的葉酸扮演了作為促進該混成物描準海拉細胞的靶向分子或配體的功能。於光動力療法中,使用介於700-1100 nm範圍內的近紅外光是特別引人注目的,因為生物系統大都缺少吸收近紅外光的發色團(chromophores),因此近紅外光對於人類身體而言較紫外光/可見光更為安全。
以GO/DEN-OH/FA混成物培養海拉細胞,且以780 nm的雷射光束照射海拉細胞。圖6顯示照射前以及照射後的共軛焦顯微影像。在圖6中,列(a)的影像是由對照組(僅有海拉細胞)照射前以及照射後所獲得,而列(b)的影像是由以GO/DEN-OH/FA混成物(DEN-OH為90 μM)培養之海拉細胞於照射前以及以780 nm照射15分鐘後所獲得。對
照組不受雷射照射的影響,而光活化GO/DEN-OH/FA混成物之存在卻導致海拉細胞的死亡。細胞死亡率會隨著混成物中DEN-OH的濃度增加而變得更高。
GO/DEN-OH/FA混成物、GO以及DEN-OH的毒性是使用MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑嗅鹽,黃色的四唑)試驗法來評估,其結果如圖7所表示。圖7顯示細胞存活率取決於GO(●)、DEN-OH(▲)以及GO/DEN-OH/FA混成物(■)的濃度。結果指出,當GO、DEN-OH以及GO/DEN-OH/FA混成物相對於海拉細胞的濃度增加,細胞的存活率下降。不過,在GO/DEN-OH/FA混成物的存在下觀察到海拉細胞的存活率是足夠高的。所培養之細胞在GO/DEN-OH/FA混成物的濃度高達20 ug/ml時展現約80%的存活率,指出GO/DEN-OH/FA混成物的低細胞毒性。
受激發的光感物質可藉由放射光線(螢光)或內轉換成熱的方式失去能量。然而,受激發之光感物質可能經歷系統間跨越過程(intersystem crossing process)並且進入激發三重態(excited triplet state)。三重激發態的光感物質可以將其能量轉移至鄰近的基團或分子上並導致ROS的產生。產生的ROS會氧化細胞結構,導致細胞損傷或死亡。
為了提升GO/DEN-OH/FA混成物於光動力療法的潛力,GO/DEN-OH/FA混成物依據633 nm雷射照射產生ROS
的能力可使用具N,N’-二甲基-4-亞硝基苯胺(RNO)的GO/DEN-OH/FA混成物水溶液來檢查。自由基氧化導致RNO的褪色。因為RNO的褪色使得440 nm的強吸收峰(ε=3.44×104
M-1
cm-1
)可以隨後使用紫外光可見光譜儀定量。RNO位於440 nm的收吸峰主要由環基團至亞硝基基團的π→π*
電荷轉移所貢獻。
根據GO/DEN-OH/FA混成物溶液的紫外光可見光光譜,RNO的褪色情形幾乎是相對於照射時間增加而線性地增加。因此,GO/DEN-OH/FA混成物能夠吸收近紅外光並產生ROS,ROS可以氧化海拉細胞的胞器以及細胞結構並造成海拉細胞的死亡。GO/DEN-OH/FA混成材料可作用為光感物質,導致光動力療法中標靶細胞的毀滅。
綜上所述,已合成之GO/DEN-OH/FA混成物顯示出雙光子吸收性,且此混成物於390 nm以及780 nm的激發波長表現出於450 nm的可見光區域的強烈螢光放射。GO/DEN-OH/FA混成物也具有因雷射照射而產生活性含氧物(ROS)的能力。GO/DEN-OH/FA混成物的螢光強度是基於雷射照射而顯著地增強且持續2小時不會觀察到螢光強度的消減。與GO/DEN-OH/FA混成物培養的海拉細胞以780 nm的雷射照射後,可觀察到海拉細胞的損傷或死亡(包含氣泡形成、收縮以及外觀的不平整)。在GO/DEN-OH/FA混成物含量小於20 μg/ml時,表現了令人滿意的低細胞毒性以及高生物相容性。這些GO/DEN-OH/FA的奈米混成材料可作為組織治療應用上
的螢光標記探針以及作為用於癌症療程之光動力療法中的光感物質。
本發明提供雙功能GO/DEN-OH/FA混成材料。對醫學及/或治療應用上,GO/DEN-OH/FA混成材料可作用為螢光標記或螢光探針以及作用為攻擊製劑(藥物)。意即,由於混成材料本身同時作用為螢光標記或螢光探針以及攻擊試劑(藥物)而不需額外的治療製劑,因此螢光GO/DEN-OH/FA混成材料本身就是完整的藥物傳遞系統。
由於螢光GO/DEN-OH/FA混成材料不需使用螢光標記或標籤,因此在細胞毒性以及水溶性可進一步強化。因為強烈的螢光放射,混成材料也可作用為用於螢光顯影的顯影劑。此外,可使用GO/DEN-OH/FA混成材料作為其他治療性製劑的藥物傳遞系統。
雖然本發明已以實施例揭露如上,然其並非用以限定本發明,任何所屬技術領域中具有通常知識者,在不脫離本發明之精神和範圍內,當可作些許之更動與潤飾。說明不一定是依比例繪製。本揭露之範例物件與實際裝置間可能因為製造流程以及誤差度而有所區別。本發明可具有其他未特別說明的實驗例。說明書以及圖示是用以說明而非限制本發明。可進行適當的修飾以使特定的情況、材料、物的組成、方法或物件的製程符合本發明的精神及範圍。上述的修飾是認定為後述之申請專利範圍的範圍內。於此揭露之方法描述雖然是參考特定順序進行特定操作,然而,須了解的是,這些操作可以在不背離本發明的教示之
下合併、分離或是重新排序以形成均等方法。
圖1繪示根據本發明一實施例之端羥基PAMAM樹枝狀高分子(DEN-OH)以及葉酸(FA)鍵結於石墨烯氧化物(GO)上的反應流程圖。
圖2繪示具有不同DEN-OH濃度的GO/DEN-OH/FA混成物的紫外光可見光吸收光譜。
圖3A至圖3E繪示GO/DEN-OH/FA混成物於不同激發波長的螢光發射光譜。
圖4繪示GO/DEN-OH/FA混成物(DEN-OH為90 μM)於不同激發波長的以雙光子激發的螢光影像。
圖5繪示繪示以激發波長介於720-900 nm的雙光子激發的可見的螢光GO/DEN-OH/FA混成物的數量。
圖6繪示以GO/DEN-OH/FA混成物培養的海拉細胞以及其對照組於雷射照射前以及雷射照射後的顯微影像。
圖7繪示細胞存活率與GO、DEN-OH以及GO/DEN-OH/FA混成物之濃度的影響。
Claims (10)
- 一種混成材料,包括50-256重量份的端羥基聚(醯胺基胺)樹枝狀高分子,5-20重量份的石墨烯氧化物以及0.5-16重量份的葉酸,其中該端羥基聚(醯胺基胺)樹枝狀高分子以及該葉酸化學鍵結至該石墨烯氧化物上以形成該混成材料,其中製造該混成材料是藉由混合該端羥基聚(醯胺基胺)樹枝狀高分子以及該石墨烯氧化物且於室溫中攪拌,添加N,N’-二環己基碳二亞胺(DCC)以及4-二甲基氨基吡碇(DAMP)至該端羥基聚(醯胺基胺)樹枝狀高分子與該石墨烯氧化物的混合物中且於室溫中攪拌,以進行酯化作用以形成中間產物GO/DEN-OH,GO/DEN-OH是指端羥基聚(醯胺基胺)樹枝狀高分子鍵結於石墨烯氧化物上,添加N-羥基丁二醯亞胺(NHS)以及1-(3-二甲氨基丙基)-3-乙基碳二亞胺鹽酸鹽(EDC)至該中間產物GO/DEN-OH之水性懸浮液中,添加葉酸至NHS、EDC與該中間產物GO/DEN-OH之該水性懸浮液的混合物中,且於室溫中攪拌以形成該混合材料。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料顯現出雙光子吸收性,該雙光子吸收性是藉由第一波長的螢光放射來展現,所述的第一波長的螢光放射是受到短於該第一波長的第二波長所激發以及受到長於該第一波長的第三波長所激發而產生。
- 如申請專利範圍第2項所述之混成材料,其中該第一波長為450nm,該第二波長為390nm且該第三波長為780nm。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料吸收近紅外光光線並在照光後產生活性含氧物(ROS)。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料為水可分散性。
- 如申請專利範圍第1項所述之混成材料,其中該端羥基聚(醯胺基胺)樹枝狀高分子為***端羥基聚(醯胺基胺)樹枝狀高分子。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料作用為螢光標記。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料作用為光動力療法的攻擊藥物。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料的該端羥基聚(醯胺基胺)樹枝狀高分子的濃度為18μM時,該混成材料的該石墨烯氧化物:該端羥基聚(醯胺基胺)樹枝狀高分子:該葉酸的相對重量比例為11:50:9。
- 如申請專利範圍第1項所述之混成材料,其中該混成材料的該端羥基聚(醯胺基胺)樹枝狀高分子的濃度為90μM時,該混成材料的該石墨烯氧化物:該端羥基聚(醯胺基胺)樹枝狀高分子:該葉酸的相對重量比例為11:256:9。
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US20120177593A1 (en) * | 2009-07-20 | 2012-07-12 | The Regents Of The University Of Michigan | Synthesis of dendrimer conjugates |
CN101670108A (zh) * | 2009-08-13 | 2010-03-17 | 苏州纳米技术与纳米仿生研究所 | 基于纳米氧化石墨烯的载药体系 |
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US8927658B2 (en) | 2015-01-06 |
US20140080980A1 (en) | 2014-03-20 |
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