200410729 玖、發明說明 、實施方式及圖式簡單說明) (發明說明應敘明：發明所屬之技術領域、先前技術、內容 一、發明所屬之技術領域 本發明係關於一種多層傳輸系統，尤指一種適用於 制酸同時增加腸道黏膜吸收之多層傳輸系統。 二、先前技術 奴著生物科技的蓬勃發展，生技藥物如胜肽等，已 經成為目前藥物發展的主流趨勢，但是生技藥物由於分 子較大且安定性較一般小分子藥物為差，易受到環境的 破壞’因此目前的給藥途徑皆以注射為主。對大分子藥 物而言，皮T注射比口服傳遞的吸收率增加了將近16〇〇 倍’這是因為-般胜肽/生技藥物以口服傳遞時會被消 化，中之胃酸與酵素分解，而使得藥物失效，且胜月太/生 技藥物皆為大分子’不易被腸胃道吸收，因此多以注射 方式進行，對病人而言較為痛苦。而針劑的製造過程較 為繁瑣，因其必須經過嚴格的滅菌過程，成本亦較高，。 反之’ 口服劑型為非侵入性之治療方式，可減少病人的 不便。因此以-般藥物而言，目前口服劑型的佔有率約 為53%左右，使用上更是注射劑型（1〇%)的五倍，顯示口 服：型仍是最普遍使用的給藥方式。目前生技藥物約五 十夕種’有更多的藥物在臨床試驗中，隨著生物科技的 進步’陸續將有—百多種生技醫藥品上市，口服胜肽/生 技藥物劑型之研發將使得此類生技藥品的❹率 遍，進而嘉惠更多病人。 曰 ，脂粒（liP〇s〇me)以及高分子載體均被認為是傳遞 生技藥物的最佳載體。微脂粒以磷脂質雙層結構内包含 办ί·£^· 6 200410729200410729 (Instructions of the Invention, Brief Description of the Embodiments and the Drawings) (The description of the invention should state: the technical field to which the invention belongs, the prior art, and the content 1. The technical field to which the invention belongs The present invention relates to a multilayer transmission system, especially a It is suitable for multi-layer transmission system for acid production and increase absorption of intestinal mucosa. 2. The previous technology slaved to the vigorous development of biotechnology. Biotechnology drugs such as peptides have become the mainstream trend of current drug development. Larger and less stable than ordinary small-molecule drugs, vulnerable to environmental damage ', so the current route of administration is mainly injection. For large-molecule drugs, the absorption rate of dermal T injection has increased by nearly than that of oral delivery. 1600 times 'This is because-peptides / biotech drugs are digested when delivered orally, and the stomach acid and enzymes are broken down, which makes the drug ineffective, and it is not easy for Katsuetai / biotech drugs to be macromolecules' It is absorbed by the gastrointestinal tract, so it is mostly performed by injection, which is more painful for the patient. The manufacturing process of the injection is more complicated, because it must be After a strict sterilization process, the cost is also higher. Conversely, the oral dosage form is a non-invasive treatment method that can reduce the inconvenience of the patient. Therefore, as for ordinary drugs, the current oral dosage form has a share of about 53%. The use is five times that of the injection form (10%), showing that the oral: type is still the most commonly used method of administration. At present, about fifty Chinese medicine is a biotech drug. There are more drugs in clinical trials. With the advancement of biotechnology, there will be more than one hundred kinds of biopharmaceutical products on the market, and the development of oral peptides / biopharmaceutical formulations will make the prevalence of such biopharmaceuticals more widespread, thus benefiting more patients. Lipids (liP0some) and polymer carriers are considered to be the best carriers for delivering biopharmaceuticals. Microlipids contain phospholipid double-layered structures containing 办 · ^^ 6 200410729
一藥劑活性成分，以達到高包覆率的目標。然而一般微 脂粒以口服方式傳遞藥物時，易受到腸胃道的酵素等破 壞，因此需要對微脂粒的外層做修飾，以延長微脂粒在 體内循環的時間。目前較為可行的方式係以多層傳輸系 統的方法，期使外層的物質可以抵抗腸胃道的酸、鹼性 酵素。 目前微脂粒在此方面的技術是以Generex、MIT以及 Oralsomal Technology公司為代表，而高分子載體則是以 Adavanced Polymers Systems，Inc為主。其中MIT以及 Oralsomal Technology是屬於同一技術來源，由Robert Langer之研發團對所開發，其貯藥目的是將微脂粒標的 至腸胃細胞，因其使用配方包覆生技藥物的效率不高， 因此較為適合作用於如疫苗等potency較高的藥物。而 Generex 則是以加入 Triolein 及 SLS ( sodium laurylsulphate)等作為促進吸收劑，並且以oral spray方 式服用，然而由於以口腔黏膜作為吸收途徑，其吸收面 積遠較腸胃道為少，因此需要使用大量的促進吸收劑， 方能達到一定的藥效，但生技藥物多為需長期服用之藥 物，因此其劑型之毒性亦較高。Advanced Polymers Systems Inc.之專利主要是以Aliginate/Chitosan系統作為 生技藥物的載體，由於僅使用Alginate/Chitosan系統包覆 生技藥物，不但藥物的吸收不佳，更由於使用擴散的方 式包覆蛋白質，其包覆率亦不高，因此其生技藥物絕大 部分並未包覆於Alginate/Chitosan微粒中，而即使藥物可 7 200410729 於腸胃道中釋放，但生技筚 易穿、# …物鐵會被酵素所分解並且不 易穿透細胞，效果十分有限。 个 二、發明内容 傳要目的係在提供-歡物活性成分多層 間使該多層傳輸系統在腸胃道中的停滯時 1B田活性成分（例如藥物）包覆率提高，同時A pharmaceutical active ingredient to achieve the goal of high coverage. However, in general, when liposomes are used to deliver drugs orally, they are easily damaged by enzymes such as the gastrointestinal tract. Therefore, the outer layer of the liposomes needs to be modified to prolong the circulation time of the liposomes in the body. At present, a more feasible method is to use a multi-layer transmission system, so that the outer layer can resist the acid and alkaline enzymes in the gastrointestinal tract. At present, the technology of liposomes in this regard is represented by Generex, MIT and Oralsomal Technology, and the polymer carrier is mainly Adavanced Polymers Systems, Inc. Among them, MIT and Oralsomal Technology belong to the same technology source and were developed by the research team of Robert Langer. The purpose of the drug storage is to target the microlipids to the gastrointestinal cells. Because the efficiency of using the formula to coat biotechnology drugs is not high, therefore It is more suitable for drugs with higher potency such as vaccines. Generex is supplemented with Triolein and SLS (sodium laurylsulphate) as absorption enhancers, and is taken by oral spray. However, as the oral mucosa is used as an absorption route, its absorption area is much smaller than that of the gastrointestinal tract, so a large amount of Promoting the absorption agent can achieve a certain effect, but most biotechnology drugs are drugs that need to be taken for a long time, so the toxicity of their dosage forms is also high. The patent of Advanced Polymers Systems Inc. is mainly based on the Aliginate / Chitosan system as the carrier of biopharmaceuticals. Since only the Alginate / Chitosan system is used to coat the biopharmaceuticals, not only the absorption of the drug is poor, but also the protein is coated by diffusion. , Its coating rate is not high, so most of its biotechnology drugs are not coated in Alginate / Chitosan microparticles, and even though the drug can be released in the gastrointestinal tract 7 200410729, biotechnology is easy to wear, #… It is broken down by enzymes and does not easily penetrate cells. The effect is very limited. Second, the purpose of the invention is to provide the active ingredient of the fungal substance to make the multilayer delivery system stagnate in the gastrointestinal tract. The coverage of active ingredients (such as drugs) in the field is improved at the same time.
=抑制肪月道酵素對於藥物的作用，達到Π服傳輸的效 果0 本^月之另-目的係在提供_種生物活性成分多層 專輸糸統的製備方法，俾能製備_種生物活性成分多層 傳輸系統’使多層傳輸系統在腸胃道中的停滯時間延 長，生物活性成分（例如藥物）包覆率提高，同時能抑 制腸胃道酵素對於藥物的作用，達到口服傳輸的效果。 為達成上述目的，本發明之多層傳輸系統之製備方 法’主要包括：提供-微脂粒溶液，該微脂粒中包覆有 一生物活性成分；提供一包含有至少一多孔性高分子成 分之包覆溶液，並與該微脂粒溶液混合；以及將步驟（2) 中之$混合液喷至-具有至少_交聯劑或具黏膜沾黏性高 分子聚合物之溶液中，交聯後並過濾分離，得到一多層 包覆傳輸系統；其中，至少一高分子成分係選自一由= Inhibit the effect of fatty enzymes on drugs and achieve the effect of delivery. 0 Another month-the purpose is to provide a method of preparing _ species of biologically active ingredients, a multi-layered delivery system, which cannot produce _ kinds of biologically active ingredients. The multi-layered delivery system 'prolongs the stagnation time of the multi-layered delivery system in the gastrointestinal tract, improves the coverage of bioactive components (such as drugs), and can inhibit the effect of gastrointestinal enzymes on drugs, achieving the effect of oral delivery. In order to achieve the above-mentioned object, the preparation method of the multilayer transmission system of the present invention mainly includes: providing a microlipid solution, the microlipid particle is coated with a biologically active ingredient; and providing a microlipid solution containing at least one porous polymer component. Coating the solution and mixing it with the microlipid solution; and spraying the $ mixed solution in step (2) to a solution having at least a cross-linking agent or a mucous membrane-adhesive polymer, after crosslinking And filtering and separating to obtain a multilayer coating transmission system; wherein at least one polymer component is selected from a group consisting of
Sodium Alginate、Alginic Acid、Pr0pylene GlycolSodium Alginate, Alginic Acid, Pr0pylene Glycol
Alginate、Gelatin、Chitosan、Hydroxy Propyl Cellulose、 Hydroxy Propyl Methyl Cellulose 以及 P〇iyacrylates、以及 Dextran所組成之群組；且該交聯液所包含之該高分子聚 8 200410729 合物係選自一由 Chitosan、Polyacrylates、Dextran、 Hydroxy Propyl Cellulose、Hydroxy Propyl Methyl Cellulose以及Gelatin所組成之群組。 本發明同時提供一種多層傳輸系統，包含：至少一 微脂粒；至少一多孔成分之親水性高分子包覆層；以及 具有高分子聚合物之黏膜吸收層其中該包覆層係介於該 微脂粒以及該黏膜吸收層之間；該微脂粒主要包括一磷 脂質雙層包覆外層、一水性核心，以及一位於該水性核 心中之一生物活性物質；該包覆層之成分係選自一由 Sodium Alginate、Alginic Acid、Propylene Glycol Alginate、Gelatin、Chitosan、Hydroxy Propyl Cellulose、 Hydroxy Propyl Methyl Cellulose以及 Polyacrylates、所組 成之群組；該黏膜吸收層之高分子聚合物係選自一由 Chitosan、Polyacrylates、Dextran、Hydroxy Propyl Cellulose、Hydroxy Propyl Methyl Cellulose 以及 Gelatin 所組成之群組。 藉由本發明所提供之生物活性成分多層傳輸系統及 其製造方法，可得到一種生物活性成分多層傳輸系統， 除了在中心微脂粒中包覆有一生物活性物質之外，更包 含有一制酸層以抑制腸胃道中之酸性酵素，同時亦包含 有一黏膜吸收層，可有效增加該多層包覆微脂粒在腸道 黏膜中被吸收之效率。 本發明之製備方法中，該微脂粒溶液較佳係由以下 步驟製備：先將一藥劑活性物質溶於一鹽類溶液中，接 著將該鹽類溶液加入一預先混合之脂質中，攪拌成一混 9 200410729 合液；以及該混合液，並將該混和合液加入一酸溶液中， 水合後得到微脂粒溶液。本發明製備方法中，該預先混 合之脂質較佳係至少一選自一由磷脂質 (phospholipids)、distearylphosphatidylethanolamine mPEG2000、distearylphosphatidylethanolamine mPEG5000、fatty acids、triglycerides、diglycerides、 monoglycerides、polyoxylethylene alkyl ethers、 polyoxylethylene glyceryl fatty acid esters、bile salts、維 他命E及其衍生物、膽固醇以及正電荷脂質所組成之群 組；其中該填脂質較佳係為大豆填脂膽鹼（soybean phosphatidylcholine );該 polyoxylethylene alkyl ethers 較佳係為 polyoxyethylene (23) lauryl ether ;該維他命 E及 其衍生物較佳係為 a -Tocopherol Succinate PEG 1500 ;該 正電荷脂質較佳係為Steaylamine ;該polyoxylethylene alkyl ethers較佳係為 p〇ly〇xyethylene (23) lauryl ether;該 大豆構脂膽驗、polyoxyethylene (23) lauryl ether與膽固 醇二者之莫耳比例並無限制，較佳為 0·5_2:0·05-0·5:0.05-0.5 ;該大豆磷脂膽鹼、a -Tocopherol Succinate PEG 1500與膽固醇三者之莫耳比例並無限 制，較佳為0.5-2:0.05-0.5:0.05-0.5 ;該正電荷脂質係為 Steaylamine ;該大豆填脂膽鹼、p〇iyOXyethylene (23) lauryl ether及stearylamine與膽固醇該四者之莫耳比例 並無限制’較佳為 0.5-2:0.05-0.5:0.05-0.5:0.05-0.5 ;該包 覆溶液中之多孔性成分較佳為藻酸納（s〇dium alginate); 該高分子聚合物較佳為幾丁聚醣（chitosan);藻酸鈉的濃 200410729 度較佳為1_20% ;該交聯液中較佳含有〇·5-5%氣化約以及 0.5-3%幾丁聚醣；該包覆溶液與該微脂粒溶液二者之體 積比並無限制，然該包覆溶液與該微脂粒溶液之比例較 佳為卜5:1 ;該包覆溶液中之多孔性成分較佳為藻酸 納；該PolyAcrylates高分子較佳為cabopol 971NF，Cabopol 934NF或前二者之混合物；該藻 酉夂納之ί辰度較佳為1-20%，P〇lyacrlates之濃度較佳為 1-20%，該交聯液中較佳係含有〇.5_5%氯化鈣；本 發明之製備方法中該包覆溶液與該微脂粒溶液二 者之體積比並無限制，較佳為1 _ 5 : 1 ;該包覆溶 液中之多孔性成分較佳為藻酸鈉；該藻酸鈉的濃度較 佳為1-20%，Polyacrlates之濃度較佳為1-20%，交聯液中 較佳含有0.5-5%氣化鈣以及0.5-3%幾丁聚醣；該包覆溶 液中之多孔成分較佳為藻酸鈉及Dextran ;該交 聯液中較佳含有PEG 20_50%以及氯化約i-i〇〇/0。 四、實施方式 為能讓貴審查委員能更瞭解本發明之技術内容，特 舉八較佳具體實施例說明如下。 胰島素微脂粒的製備 貫施例一 秤取0.5g之脂質於15mL玻璃試管中，其中soybeail phosphatidylcholine - polyoxyethylene (23) lauryl ethers 膽固醇之莫耳比例為1 :〇· 1 ··〇· 1;同時取適量的人類胰島素 11 200410729 以0·01Μ，ρΗ7·4之磷酸溶液溶解後，取〇.5mL，70mg/mL的 人類胰島素溶液加入上述的脂質中。使用Vortex混合後， 置以200rpm攪拌一小時候，取0.2g之黏稠膏狀物於10mL 的燒杯中，加入1·8 mL，pH 7·4之磷酸溶液，於室溫下進 行1小時的水合，即得到胰島素微脂粒溶液。微脂粒中的 胰島素達到1.86mg/mL，包覆率達58%以上。 實施例二 秤取0.5g之脂質於15mL玻璃試管中，其中soybean phosphatidylcholine、a -Tocopherol Succinate PEG 1500 及膽固醇之莫耳比例為1:0.1:0.1 ;同時取適量的人類胰島 素以0.01M，ρΗ7·4之磷酸溶液溶解後，取0·5 mL，60mg/mL 的人類胰島素溶液加入上述的脂質中。使用Vortex混合 後，置以200rpm攪:拌一小時候，取0.2g之黏稠膏狀物於 10mL的燒杯中，加入1.8 mL，pH 7.4之填酸溶液，於室溫 下進行1小時的水合，即得到胰島素微脂粒溶液。微脂粒 中的胰島素達到1.87mg/mL，包覆率達63%以上。 實施例三 秤取0.5g之脂質於15mL玻璃試管中，其中soybean phosphatidylcholine、stearylamine及膽固醇之莫耳比例為 1:0.1:0.1 ;同時取適量的人類胰島素以0.01M，ρΗ2·0之稀 鹽酸溶液溶解後，取0.5 mL，60mg/mL的人類胰島素溶液 加入上述的脂質中。使用Vortex混合後’置以200rpm攪 拌一小時候，取〇.2g之黏稠膏狀物於l〇mL的燒杯中，加 系·ίν·ί4·‘ 12 200410729 入0.8 mL，pH 2.0之磷酸溶液，於室溫下進行1小時的水 合，即得到胰島素微脂粒溶液。微脂粒中的胰島素達到 2.9mg/mL，包覆率達60%以上。 膜島素微脂粒微粒包覆（以spray Nozzle方法） 實施例四 秤取適量的藻酸鈉粉末，加入ρΗ7·4磷酸溶液使得藻 酸鈉的重量濃度達到10%。攪拌至完全溶解後，將溶液中 的氣泡除去，接者取實施例一中之胰島素微脂粒溶液與 藻酸鈉溶液以等比例的體積混合後攪拌均勻。使用 Turbosonic之2mm喷嘴將藻酸鈉與胰島素微脂粒混合溶 液以lmL/min的流速以及1.5kg/cm2的壓力將溶液喷至含 有1.5%氯化#5及0.8%幾丁聚醣（chitosan)的溶液中進行交 聯。將溶液過濾後得到胰島素微脂粒微粒，包覆率為 37.6%。 實施例五 φ 秤取適量的藻酸鈉粉末，加入p Η 2的鹽酸溶液使得藻 酸鈉的重量濃度達到10%。攪拌至完全溶解後，將溶液中 的氣泡除去，接者取實施例三中之胰島素微脂粒溶液與 藻酸鈉溶液以2:1的體積混合後攪拌均勻。使用 Turbosonic之2mm喷嘴將藻酸鈉與胰島素微脂粒混合溶 液以lmL/min的流速以及1.5kg/cm2的壓力將溶液喷至含 有1.5%氣化#5及0.8%幾丁聚醣（chitosan)的溶液中進行交 聯。將溶液過濾後得到胰島素微脂粒微粒，包覆率達76%。 13 200410729 實施例六 秤取適量的Cabopol 971與藻酸鈉粉末，加入去離子 水並攪拌，使得Cabopol 971與藻酸鈉完全溶解後，將溶 液中的氣泡除去，接者取實施例一中之胰島素微脂粒溶 液以pHl.O之鹽酸調整pH值至3.7後，與上述高分子溶液 溶液以2:1的重量比例混合後攪拌均勻。使用Turbosonic 之1mm喷嘴將混合溶液以lmL/min的流速以及2.0kg/cm2 的壓力將溶液喷至含有5%氣化鈣的溶液中進行交聯。將 溶液過滤後付到膜島素微脂粒微粒，包覆率達9 0 %。 胰島素微脂粒微粒包覆（以Two Aqueous Phase Preparation ) 實施例七 將lg dextran及〇.25g藻酸鈉（LVCR)溶解於實施例一 中之胰島素微脂粒溶液（hydration 1:4, 3.75g)中，待其完 全溶解後，將lg dextran/藻酸鈉/微脂粒溶液加入2g PEG 溶液中（10% PEG 10k，0.06% NaCl)，並以磁石攪拌30分 鐘以形成乳化液。接著以滴管緩慢加入1 g之氯化#5溶液 (PEG 30%，氯化鈣5%)，並持續以磁石攪拌30分鐘，以交 聯藻酸鈉而形成胰島素微脂粒微粒，製備完成後以減壓 過濾方式分離，其包覆率達90%以上。 實施例八、糖尿病鼠實驗 200410729 2〇0-；250g重之Wistar Rats以腹腔注射方式注射 65mg/Kg STZ，觀察病鼠血糖的濃度，直到血糖為直3〇〇 mg/dL以上，餵食樣品前一天晚上病鼠需禁食一晚。以儀 食針餵食約100IU/kg的樣品至老鼠胃部，每隔1小時取血 一次，利用one touch glucose meter測量其血液中的 glucose濃度。測量結果如圖1所示，可證明本發明之多層 包覆微脂粒的確具有增加腸胃道黏膜吸收及抑制胃酸破s 壞微脂粒之功效。 由以上之實施例以及說明可以知道，本發明之生物 活性成分多層傳輸系統較諸先前技藝，可以提高生物活 性成分（例如藥物）之包覆率’並可以延長該多層傳輸 系統在腸胃道中的停滯時間延，抑制腸胃道酵素對於微 脂粒所包覆藥物的作用，而達到提高口服傳輪的效果: 上述實施例僅係為了方便說明而舉例而已，本發明 所主張之權利範圍自應以申請專利範圍所述為準 僅限於上述實施例。 五、圖式簡單說明 變化圖 圖1係本發明實施例八之糖尿病鼠之血糖對時間Alginate, Gelatin, Chitosan, Hydroxy Propyl Cellulose, Hydroxy Propyl Methyl Cellulose, Poiyacrylates, and Dextran; and the polymer 8 included in the cross-linking solution is selected from a group consisting of Chitosan, A group of Polyacrylates, Dextran, Hydroxy Propyl Cellulose, Hydroxy Propyl Methyl Cellulose, and Gelatin. The present invention also provides a multi-layer transmission system comprising: at least one microfat particle; a hydrophilic polymer coating layer of at least one porous component; and a mucosal absorption layer having a polymer polymer, wherein the coating layer is interposed between the Between the microlipid particles and the mucosal absorption layer; the microlipid particles mainly include a phospholipid double-layered outer layer, an aqueous core, and a bioactive substance located in the aqueous core; the composition of the coating layer It is selected from the group consisting of Sodium Alginate, Alginic Acid, Propylene Glycol Alginate, Gelatin, Chitosan, Hydroxy Propyl Cellulose, Hydroxy Propyl Methyl Cellulose, and Polyacrylates; the polymer of the mucosal absorption layer is selected from a group consisting of Chitosan , Polyacrylates, Dextran, Hydroxy Propyl Cellulose, Hydroxy Propyl Methyl Cellulose, and Gelatin. Through the multi-layered biologically active ingredient transmission system and manufacturing method provided by the present invention, a multi-layered biologically active ingredient transmission system can be obtained. In addition to coating a biologically active substance in the central microlipids, it also includes an acid-producing layer to Inhibits acid enzymes in the gastrointestinal tract, and also contains a mucosal absorption layer, which can effectively increase the efficiency of the multi-layer coated microlipids being absorbed in the intestinal mucosa. In the preparation method of the present invention, the microlipid solution is preferably prepared by the following steps: first dissolving a medicament active substance in a salt solution, and then adding the salt solution to a pre-mixed lipid and stirring to form a Mix 9 200410729; and the mixed solution, and add the mixed solution to an acid solution, and obtain a microlipid solution after hydration. In the preparation method of the present invention, the premixed lipid is preferably at least one selected from the group consisting of phospholipids, distearylphosphatidylethanolamine mPEG2000, distearylphosphatidylethanolamine mPEG5000, fatty acids, triglycerides, diglycerides, monoglycerides, polyoxylethylene alkyl ethers, polyoxylethylene glyceryl fatty acid esters , Bile salts, vitamin E and its derivatives, cholesterol, and positively charged lipids; wherein the lipid-filling lipids are preferably soybean phosphatidylcholine; the polyoxylethylene alkyl ethers are preferably polyoxyethylene ( 23) lauryl ether; the vitamin E and its derivatives are preferably a-Tocopherol Succinate PEG 1500; the positively charged lipid is preferably Steaylamine; the polyoxylethylene alkyl ethers are preferably p〇ly〇xyethylene (23) lauryl ether; there is no limit to the mole ratio of the soybean lipid-forming bile test, polyoxyethylene (23) lauryl ether and cholesterol, preferably 0.5-5_2: 0 · 05-0 · 5: 0.05-0.5; the soybean phospholipid bile Alkali, a -Tocopherol Succinate PEG There is no limitation on the molar ratio of 1500 to cholesterol, preferably 0.5-2: 0.05-0.5: 0.05-0.5; the positively charged lipid system is Steaylamine; the soybean fat-filling choline, pOyOXyethylene (23) lauryl There are no restrictions on the molar ratios of ether and stearylamine to cholesterol. 'It is preferably 0.5-2: 0.05-0.5: 0.05-0.5: 0.05-0.5; the porous component in the coating solution is preferably sodium alginate (S〇dium alginate); The high molecular polymer is preferably chitosan; the concentration of sodium alginate 200410729 is preferably 1-20%; the cross-linking solution preferably contains 0.5-5% Gasification is about 0.5-3% chitosan; the volume ratio of the coating solution and the microlipid solution is not limited, but the ratio of the coating solution to the microlipid solution is preferably 5 : 1; the porous component in the coating solution is preferably sodium alginate; the PolyAcrylates polymer is preferably cabopol 971NF, Cabopol 934NF or a mixture of the two; the degree of alginate is preferably 1-20%, and the concentration of Poliacrlates is preferably 1-20%, and the cross-linking solution preferably contains 0.5-5% calcium chloride; the package in the preparation method of the present invention The volume ratio of the solution and the microlipid solution is not limited, preferably 1_5: 1; the porous component in the coating solution is preferably sodium alginate; the concentration of the sodium alginate is preferably 1-20%, the concentration of Polyacrlates is preferably 1-20%, and the cross-linking solution preferably contains 0.5-5% calcium carbonate and 0.5-3% chitosan; the porous component in the coating solution is better It is sodium alginate and Dextran; the cross-linking solution preferably contains 20-50% PEG and about 2100/0 chloride. 4. Implementation Modes In order for your review committee to better understand the technical content of the present invention, eight preferred embodiments are described below. Preparation of Insulin Microlipids In Example 1, 0.5g of lipids were weighed into 15mL glass test tubes, in which the molar ratio of soybeail phosphatidylcholine-polyoxyethylene (23) lauryl ethers cholesterol was 1: 〇 · 1 ·· 〇 · 1; An appropriate amount of human insulin 11 200410729 was dissolved in a phosphoric acid solution of 0.01 M and pH 7.4, and then 0.5 mL of a 70 mg / mL human insulin solution was added to the above lipid. After mixing with Vortex, place it at 200 rpm for one hour, take 0.2 g of viscous paste into a 10 mL beaker, add 1.8 mL of a phosphoric acid solution at pH 7.4, and perform hydration at room temperature for 1 hour. The insulin microlipid solution is obtained. Insulin in microfat granules reached 1.86 mg / mL, and the coverage rate reached more than 58%. Example 2 Weigh 0.5g of lipid in a 15mL glass test tube, in which the molar ratio of soybean phosphatidylcholine, a-Tocopherol Succinate PEG 1500 and cholesterol is 1: 0.1: 0.1; at the same time, take an appropriate amount of human insulin to 0.01M, ρΗ7 · After the phosphoric acid solution of 4 was dissolved, 0.5 mL, 60 mg / mL of human insulin solution was added to the above lipid. After mixing with Vortex, stir at 200 rpm: mix for a little while, take 0.2 g of viscous paste into a 10 mL beaker, add 1.8 mL of a filled acid solution at pH 7.4, and hydrate at room temperature for 1 hour, that is, A solution of insulin liposomes was obtained. Insulin in microfat granules reached 1.87 mg / mL, and the coverage rate was over 63%. Example 3 Weigh 0.5g of lipid in a 15mL glass test tube, in which the molar ratio of soybean phosphatidylcholine, stearylamine and cholesterol is 1: 0.1: 0.1; at the same time, take an appropriate amount of human insulin in a dilute hydrochloric acid solution of 0.01M and ρΗ2.0 After dissolving, add 0.5 mL of human insulin solution at 60 mg / mL to the above lipid. After mixing with Vortex, set it at 200 rpm and stir for one hour. Take 0.2 g of the viscous paste into a 10 mL beaker, add · ίν · ί 4 '12 200410729 into 0.8 mL of a phosphoric acid solution at pH 2.0, and Hydration was performed at room temperature for 1 hour to obtain an insulin microlipid solution. Insulin in microfat granules reached 2.9 mg / mL, and the coverage rate was over 60%. Membrane microlipid particles are coated (by spray nozzle method) Example 4 Weigh an appropriate amount of sodium alginate powder and add ρΗ7.4 phosphoric acid solution to make the weight concentration of sodium alginate reach 10%. After stirring until completely dissolved, the air bubbles in the solution were removed, and then the insulin microlipid solution and the sodium alginate solution in Example 1 were mixed in an equal volume and stirred well. Using Turbosonic's 2mm nozzle, a mixed solution of sodium alginate and insulin microlipids was sprayed at a flow rate of 1 mL / min and a pressure of 1.5 kg / cm2 to a solution containing 1.5% chlorinated # 5 and 0.8% chitosan. The solution is crosslinked. The solution was filtered to obtain insulin microlipid particles with a coating ratio of 37.6%. Example 5 Weigh an appropriate amount of sodium alginate powder and add a hydrochloric acid solution of p p 2 to make the weight concentration of sodium alginate reach 10%. After stirring until completely dissolved, the air bubbles in the solution were removed, and then the insulin microlipid solution and the sodium alginate solution in Example 3 were mixed in a volume of 2: 1 and stirred well. Using Turbosonic's 2mm nozzle, a mixed solution of sodium alginate and insulin microlipids was sprayed at a flow rate of 1 mL / min and a pressure of 1.5 kg / cm2 to a solution containing 1.5% gasification # 5 and 0.8% chitosan. The solution is crosslinked. After filtering the solution, insulin microlipid particles were obtained, and the coating ratio reached 76%. 13 200410729 Example 6 Weigh the appropriate amount of Cabopol 971 and sodium alginate powder, add deionized water and stir to completely dissolve Cabopol 971 and sodium alginate, then remove the air bubbles in the solution, then take the one in Example 1. After the insulin microlipid solution was adjusted to pH 3.7 with hydrochloric acid of pH 1.0, it was mixed with the above polymer solution at a weight ratio of 2: 1 and stirred well. Using a Turbosonic 1mm nozzle, the mixed solution was sprayed into a solution containing 5% calcium carbonate at a flow rate of 1 mL / min and a pressure of 2.0 kg / cm2 for crosslinking. The solution was filtered and applied to the membrane island microlipid particles, with a coating ratio of 90%. Insulin microlipid microparticles coated (with Two Aqueous Phase Preparation) Example 7 Ig dextran and 0.25g sodium alginate (LVCR) were dissolved in the insulin microlipid solution (hydration 1: 4, 3.75g) ), After it is completely dissolved, add lg dextran / sodium alginate / microlipid solution to 2g PEG solution (10% PEG 10k, 0.06% NaCl), and stir with a magnet for 30 minutes to form an emulsion. Then slowly add 1 g of chlorinated # 5 solution (PEG 30%, calcium chloride 5%) with a dropper, and continue stirring with a magnet for 30 minutes to cross-link sodium alginate to form insulin microlipid particles. The preparation is complete It is then separated by vacuum filtration, and its coating ratio reaches more than 90%. Example 8: Diabetic rat experiment 200410729 2000-200; 250g Wistar Rats were injected with 65mg / Kg STZ by intraperitoneal injection, and the blood glucose concentration of the sick mice was observed until the blood glucose was more than 300mg / dL. Before feeding the samples Sick rats need to fast one night. A sample of about 100 IU / kg was fed to the rat's stomach with an instrument needle, and blood was collected every 1 hour, and the glucose concentration in the blood was measured using a one touch glucose meter. The measurement results are shown in FIG. 1, which can prove that the multi-layer coated liposomes of the present invention indeed have the effects of increasing gastrointestinal mucosal absorption and inhibiting gastric acid breakdown. From the above examples and descriptions, it can be known that the multi-layered biologically active ingredient delivery system of the present invention can improve the coverage rate of bioactive ingredients (such as drugs) compared to the prior art, and can prolong the stagnation of the multilayered delivery system in the gastrointestinal tract Time delay, inhibiting the effect of gastrointestinal enzymes on microlipid-coated drugs, and improving the effect of oral transmission: the above examples are merely examples for the convenience of explanation, and the scope of rights claimed by the present invention should be applied for The scope of the patent is subject to the foregoing embodiments. V. Brief description of the diagram Change diagram FIG. 1 shows the blood glucose versus time of a diabetic rat according to the eighth embodiment of the present invention.
1515