200403078 玖、發明說明： L發明所屬之技術領域3 本發明涉及使天然大麻素化合物，特別是Δ9-四氫大 麻酚(THC)穩定的藥物製劑。本發明進一步涉及製備該製 5 劑的方法。 【先前技術3 天然大麻素化合物可以從幾種天然源獲得，但通常 從大麻（Cannabis Sativa)獲得，它可以用作許多種疾病治 療的治療劑。對於天然大麻素化合物的綜述，參見David 10 T. Brown ed.? Cannabis, Harwood Academic Publishers 1998, ISBN 90-5702-291-5。天然大麻素化合物的例子是 THC，它在市場上名爲Marinol®(普通名爲屈大麻酚）。當 前，將THC配製爲用於口服給藥的軟明膠膠囊，其中將 藥物溶於油中。缺點在於此製劑中的THC是不穩定的。 15 因此，它必須在低溫（4°C)下貯存。很明顯化合物的低穩 定性和需要在冰箱中貯存藥物製劑是藥物産品的嚴重缺 點。 本發明的目的是提供不穩定的天然大麻素化合物如 THC的製劑，該製劑以一定的方式改進化合物的穩定性 20 使得它們可以在環境條件下貯存延長的時間。此外的目 的是提供獲得乾粉狀態的藥物物質的方法。乾燥狀態使 得開發其他劑型如用於肺輸送的乾粉製劑和用於口服或 舌下給藥的片劑成爲可能。 WO9932107公開了環糊精在雙相輸送系統或微球輸 5200403078 (ii) Description of the invention: Technical field to which the invention belongs 3. The present invention relates to a pharmaceutical preparation for stabilizing a natural cannabinoid compound, particularly Δ9-tetrahydrocannabinol (THC). The invention further relates to a method for preparing the agent. [Prior Art 3 Natural cannabinoid compounds can be obtained from several natural sources, but are usually obtained from Cannabis Sativa, which can be used as a therapeutic agent for the treatment of many diseases. For a review of natural cannabinoid compounds, see David 10 T. Brown ed.? Cannabis, Harwood Academic Publishers 1998, ISBN 90-5702-291-5. An example of a natural cannabinoid compound is THC, which is marketed under the name Marinol® (commonly known as dronabinol). Currently, THC is formulated as soft gelatin capsules for oral administration in which the drug is dissolved in oil. The disadvantage is that THC in this formulation is unstable. 15 Therefore, it must be stored at low temperature (4 ° C). It is clear that the low stability of the compounds and the need to store pharmaceutical preparations in the refrigerator are serious drawbacks of pharmaceutical products. It is an object of the present invention to provide formulations of unstable natural cannabinoid compounds, such as THC, which improve the stability of the compounds in a manner such that they can be stored under ambient conditions for extended periods of time. A further object is to provide a method for obtaining a drug substance in a dry powder state. The dry state makes it possible to develop other dosage forms such as dry powder formulations for pulmonary delivery and tablets for oral or sublingual administration. WO9932107 discloses the infusion of cyclodextrin in a biphasic delivery system or microspheres 5
.Ο D 200403078 送系統中使THC增溶的用途。環糊精的增溶作用由形成 所謂的包合配合物或主賓絡合物所引起。界〇99321〇7主 題的目的是THC的增溶以促進從鼻腔吸收。在該申請中 ’又有公開關於配製的THC的穩定性的任何情況。預先不 5此推斷形成主負絡合物的穩定效應，這是由於所屬技術 領域專業人員已知這些絡合物有時具有穩定效應但在其 他情況下由於催化作用導致活性化合物的退化。此外當 應用爲鼻或肺製劑時，環糊精具有引起粘膜刺激的缺 點。特別是具有表面活性劑性能的環糊精衍生物是粘膜 10 組織的刺激劑。 W09736577描述了可用於口服輸送親脂化合物如天 然大麻素的乾燥固體脂質組合物的用途，該固體脂質組 合物除活性物質以外包括目體脂肪和翻。此組合物的 目的在於提高口服生物可利用率而不是增強活性物質的 15 穩定性。 W00078817公開了在—種低聚糖—菊粉的存在下通 過從純的水溶液乾燥蛋白質而穩定鹼性磷酸酶。在乾燥 期間’蛋自質被由處於玻璃態的無定形菊粉組成的基質 早分子包封。其中由於將蛋白質玻璃化和與其環境隔離 2〇而達到穩^。然而，驗性碟酸酶是親水化合物，它極易 /合於水並可以直接從水溶液配製。此外穩定特別涉及蛋 白質三級和四級結構的保存，它對於酶活性是重要的。 ^〇9118()91描述了非還原糖分子，特別是單糖芽如 麥芽糖醇、乳糖醇和palatinit用於保存酶如限制性核酸内 2〇〇4〇3〇78 刀酶Pst I和抗體的穩定性的用途，它們是親水化合物。 據此專利申請，可以通過酶與糖和專有緩衝劑的混 σk後空氣乾燥而製備穩定的酶。由於這些化合物不 b 乂足里溶於極性體系，此方法不能用於親脂性化合 物0注土出 牙糖醇和乳糖醇在乾燥條件下的玻璃化轉變溫度 刀別爲44C(Y. R〇〇s，Carbohydrate Research 1993，238, 3M8)和33t。 【發明内容】 現在意外地發現高度親脂性的化合物如天然大麻素 10 化合物也可以由上述機理通過引入糖玻璃或糖醇玻璃中 而得到穩定對抗氧化和異構化。此外人們發現糖玻璃技 術還導致生物利用率提高。由於天然大麻素化合物被單 分子引入，這些化合物的溶解速率將由糖玻璃的溶解速 率確定。由於糖玻璃的溶解速率遠高於天然大麻素化合 15物的溶解速率，藥物將更迅速地呈遞到吸收膜。 在第/實施方案中，本發明涉及一種藥學組成物， 勺括天然大麻素化合物和糠戒糠醇或糖混合物或糖醇混 二物㈣璃，其特徵在於天#麻素化合物作爲單分子 包封物弓丨a糖中而不形成主賓絡合物。當在糖基質中存 20在基本上每個大麻素分子的單分子包合物時’化合物就 被引入了擦玻璃中。0此< 以認爲根才康树明此實施方 案形成的輸送系統是單相輸送系統。天然A麻素分子在 糖玻璃中無規取向。與主賓絡合物如含有環糊精的配合 u,/旦溶解，在大麻素化合物與溶解的糖分子之 7 200403078 間不再有相互作用。 將大麻素化合物引入糖玻璃中會導致糖玻璃的玻璃 化轉變溫度（Tg)降低，大麻素化合物Tg消失，和大麻素 化合物溶解速率增加。此外，掃描電子顯微術可指示是 5否引入了化合物。最優選的天然大麻素化合物是thc。 爲了獲得最高穩定性，優選糖玻璃在正常環境條件 下的玻璃化轉變溫度（Tg)大於50艽並具有低的結晶傾 向。正常環境條件定義爲2〇-25°C和至多40%相對濕度。 在本發明的構架中，措辭“天然大麻素化合物，，包括 10大麻素的非天然衍生物，該衍生物可以通過天然大麻素 的衍生作用獲得並且象天然大麻素一樣是不穩定的。 在本發明的構架中，措辭糖包括多糖而措辭糖醇包 括多糖醇。本發明中優選的糖是非還原糖。非還原糖是 沒有或不能形狀反應性醛或酮基團的糖。非還原糖的例 15 子疋海澡糖和果聚糖如菊粉。 用於本發明的優選的非還原糖是果聚糖或果聚糖混 合物。果聚糖理解爲表示包含多個脫水果聚糖單元的低 聚糖或多糖。果聚糖可具有多分散鏈長度分佈，並可含 有直鏈或支鏈。優選果聚糖主要包含卜^鍵，如在菊粉 2〇中那樣但匕們也可包含β_2,6鍵，如在左聚糖中那樣。 合適的果聚糖可直接源自天然來源，但也可經歷修飾。 修飾的例子是導致鏈長度伸長或縮短的自身已知的反 應。除天然多糖之外，4製備的多糖如鏈縮短的水解 產物和鏈長度經㈣的分級産物在本發明中也是合適 200403078 的。用於獲得鏈長度降低的果聚糖的水解反應可以通過 酶促方式（例如採用内菊粉酶），化學方式（例如採用酸的 水溶液），物理方式（例如用熱的方法）或通過使用多相催 化（例如採用酸離子交換劑）進行。果聚糖如菊粉的分級尤 5 其可以通過在低溫下結晶、採用柱色譜法分離、膜濾法 和採用醇進行選擇沈殿而實現。其他果聚糖，如長鏈果 聚糖可以例如通過從其中已經除去單糖和二糖的果聚糖 結晶而獲得。鏈長度經酶促伸長的果聚糖也可作爲本發 明的果聚糖。此外，可以使用還原的果聚糖，它們是其 10 還原性端基（通常是果糖基團）已經被還原的果聚糖，例如 採用氫硼化鈉，或用氫氣在過渡金屬催化劑存在下還 原。也可以使用經化學修飾的果聚糖，如交聯的果聚糖 和羥烧基化的果聚糖。在所有這些果聚糖中平均鏈長度 表示爲數均聚合度(DP)。縮寫DP定義爲在低聚物或聚合 15 物中糖單元的平均數目。 本發明中甚至更優選的還原糖是菊粉或菊粉混合 物。菊粉是由β-1，2結合的果糖單元與在分子還原端的 a-D-n比喃型葡萄糖單元組成的低聚糖或多糖，並可以具 有不同的聚合度(DP)。優選的菊粉是DP大於6的菊粉或其 20 中每種菊粉的DP大於6的菊粉混合物。甚至更優選DP爲 10-30的菊粉或菊粉混合物。最優選DP爲15-25的菊粉或 菊粉混合物。菊粉特別存在於百合科和菊科植物的根和 塊莖中。生産菊粉的最重要來源是洋薑，大麗花和菊苣 根。工業生産主要以菊苣根爲原料。源自不同天然來源 9 200403078 =粉之間的主要區別在於聚合度(Dp)，它可以不同， 攸在洋薑♦約6到在菊苣根中1〇_ .. Lv e 不在大麗化中尚於 。莉k低聚糖或多糖，處於無定形狀態時具有有利 _理化學性能，可用作藥物製劑中的輔助物質。這此 勿理化學性能是：（可調節的）高玻璃化轉變溫度，沒有還 原路基和通常低的結晶率。此外菊粉是無毒的並且不貴。 天然大麻素化合物對糖或糖醇的重量 W，更優壯他㈣，最優選心⑶。 ίο 15 20 本發明的藥學組成物可以進一步加工成片劑如普通 口服片劑、舌下片劑、含片或口服崩解或溶解的片劑、 膠囊、錠劑、灌腸劑、栓劑、用於透皮給藥的産品、用 於肺給藥的粉末、或用於皮下或肌内給藥的杆狀體或縣 洋液。這些給藥形式在本領域是已知的，並且所屬技術 領域的專業人員能夠將本發明的組合物加卫成所需的給 樂形式。優選的製劑是用於口服給藥或肺給藥的製劑。 用於製備本發明的糖玻璃的適宜技術是冷綠燥。 也可以制其他賴技術如讀錢m燥和超臨 界乾燥。通料些技術製備引人天鼓麻素化合物的糖 玻璃的第一步是製備其中溶解這兩種物質的溶液。铁 而’由於糖的親水性本質和天然大麻素化合物的親脂性 本質，這些化合物難以溶於相同的溶劑。現在已經發現 此問題可輯謎料_合物轉決。水是糖和糖醇 的良好溶劑’而各種有機溶劑如醇是天然域素化合物 的良好溶劑。由於水和醇很好地混合，可能在特定水/醇 10 200403078 之比下兩種物質均會實現一定程度的溶解。 因此本發明還涉及一種藥學組成物的製備方法，該 藥學組成物包括天然大麻素化合物和糖或糖混合物的玻 璃，其中天然大麻素化合物作爲單分子包封物引入糖玻 5 璃中而不形成主賓絡合物，其特徵在於 a) 將該天然大麻素化合物溶於可溶於水的有機溶劑 而將該糖或糖混合物溶於水； b) 將溶解的大麻素化合物和溶解的糖或糖混合物相 混合以獲得足夠穩定的混合物； 10 c)將該混合物冷凍乾燥，喷霧乾燥，真空乾燥或超 臨界乾燥。 適於與糖、水和天然大麻素化合物形成穩定混合物 的有機溶劑是可與水溶混的溶劑如二甲亞砜（DMSO)、 N，N-二曱基甲醯胺（DMF)、乙腈、乙酸乙酯和低級醇。由 15 於溶劑必須通過喷霧乾燥或冷凍乾燥除去，溶劑應當還 優選在乾燥溫度下具有合理的蒸氣壓。因此優選低級 醇，定義爲心-匕醇，其中烷基鏈可以是支鏈或直鏈。更 優選的醇是c2-c4醇如乙醇、正丙醇和叔丁醇。最優選的 溶劑是叔丁醇。 20 應當選擇大麻素化合物、溶劑、水和糖或糖混合物 之間的比例以獲得足夠穩定的溶液。可以選擇性地加入 表面活性劑以提高穩定性。如果在加工時間内，如在120 分鐘，60分鐘，30分鐘或10分鐘内在溶液中不出現渾濁， 則溶液判斷爲足夠穩定。對於喷霧乾燥工藝，典型的加 11 200403078 工時間是30分鐘。對於冷凍乾燥工藝，溶液應當是澄清 的直至凍結。在此加工的典型時間是10分鐘。 在乾燥過程之後水含量優選低於3%。溶劑含量優選 低於3%。本領域技術人員很清楚乾燥所需時間可從諸如 5 樣品厚度，樣品溫度，壓力，和冷凝器溫度等參數得出。 儘管使用喷霧乾燥法製備大麻素化合物的糖玻璃使 得化合物穩定性顯著改善，但採用冷凍乾燥工藝獲得最 好的結果。因此最本發明中最優選的乾燥方法是冷凍乾 燥。 10 在冷凍乾燥工藝的第一階段中，將溶液冷凍。此第 一階段應當優選快速進行並應當降低樣品溫度至低於 Tg’，丁8'是冷康濃縮級分的溫度（參見D.L· Teagarden，Eur. J. Pharm. Sci·，15, 115-133, 2002)。低於Tg’的冷凍乾燥産 生多孔餅狀物，而高於Tg’則獲得崩塌的餅狀物。由於多 15 孔餅狀物可以更容易地加工成如用於壓片的粉末或肺輸 送的製劑，所以優選多孔餅狀物。此外，在Tg’以上進行 冷凍乾燥可導致糖的結晶。這會阻止藥物被引入玻璃， 結果使穩定作用降低。 圖式簡單說明 20 第1圖係在不同濕度下菊粉的玻璃化轉變溫度。 第2圖係凍干菊粉的吸水等溫線。 第3圖係從水-TBA溶液冷凍乾燥的，包含4.00%THC的 產物的穩定。.〇 D 200403078 Soluble THC in the delivery system. The solubilization of cyclodextrin is caused by the formation of so-called inclusion complexes or guest-guest complexes. The purpose of the subject 099321〇7 is to solubilize THC to promote absorption from the nasal cavity. In this application, there is also anything disclosed regarding the stability of the formulated THC. It is inferred in advance that the stabilizing effect of forming the main and negative complexes is due to the fact that it is known to those skilled in the art that these complexes sometimes have stabilizing effects but otherwise cause degradation of the active compounds due to catalysis. In addition, when applied as a nasal or pulmonary preparation, cyclodextrins have the disadvantage of causing mucosal irritation. In particular, cyclodextrin derivatives having surfactant properties are irritants for mucosal 10 tissues. W09736577 describes the use of dry solid lipid compositions which can be used for oral delivery of lipophilic compounds such as natural cannabinoids, which solid lipid compositions include body fat and transdermal lipids in addition to the active substance. The purpose of this composition is to increase oral bioavailability rather than enhance the stability of the active substance. W00078817 discloses the stabilization of alkaline phosphatase by drying proteins from a pure aqueous solution in the presence of an oligosaccharide, inulin. During the drying period, the 'egg' is self-encapsulated by a matrix early molecule consisting of a glassy amorphous inulin. Among them, the protein is stabilized by vitrifying and isolating it from the environment by 20%. However, zymolytic enzymes are hydrophilic compounds that are extremely easy to bind to water and can be formulated directly from aqueous solutions. In addition, stabilization is particularly concerned with the preservation of tertiary and quaternary structures of proteins, which are important for enzyme activity. ^ 〇1181 (91) describes non-reducing sugar molecules, especially monosaccharide buds such as maltitol, lactitol and palatinit for preserving enzymes such as restriction enzymes in the restriction nucleic acid 20040430 Pst I and the stability of antibodies Sexual use, they are hydrophilic compounds. According to this patent application, a stable enzyme can be prepared by mixing σk of the enzyme with sugar and a proprietary buffer and air drying. Since these compounds are not soluble in polar systems, this method cannot be used for lipophilic compounds. 0 The injection temperature of the tooth-extracting sugars and lactitol is dry. The glass transition temperature is 44C (Y. R〇〇s). Carbohydrate Research 1993, 238, 3M8) and 33t. [Summary of the Invention] It has now been unexpectedly discovered that highly lipophilic compounds such as natural cannabinoid 10 compounds can also be stabilized against oxidation and isomerization by being introduced into sugar glass or sugar alcohol glass by the above mechanism. In addition, it has been found that sugar glass technology also leads to increased bioavailability. Since natural cannabinoid compounds are introduced by a single molecule, the dissolution rate of these compounds will be determined by the dissolution rate of the sugar glass. Since the dissolution rate of sugar glass is much higher than the dissolution rate of natural cannabinoid compounds, the drug will be presented to the absorption film more quickly. In a first / embodiment, the present invention relates to a pharmaceutical composition comprising a natural cannabinoid compound and bran or furfuryl alcohol or a sugar mixture or a sugar-alcohol mixed bismuth glass, which is characterized in that the tian # hemp compound is encapsulated as a single molecule The guest bow does not form a guest-guest complex in the sugar. When a single-molecule clathrate of substantially every cannabinoid molecule is present in the sugar matrix, the compound is introduced into the glass. 0This < It is considered that the transportation system formed by Gen Caikangshu this embodiment is a single-phase transportation system. The natural A-lin molecules are randomly oriented in sugar glass. With the host-guest complexes, such as containing cyclodextrin, u, once dissolved, there is no longer any interaction between the cannabinoid compound and the dissolved sugar molecule. The introduction of a cannabinoid compound into a sugar glass causes a decrease in the glass transition temperature (Tg) of the sugar glass, the disappearance of the cannabinoid compound Tg, and an increase in the dissolution rate of the cannabinoid compound. In addition, scanning electron microscopy can indicate whether the compound was introduced. The most preferred natural cannabinoid compound is thc. In order to obtain the highest stability, it is preferred that the sugar glass has a glass transition temperature (Tg) of more than 50 ° F under normal environmental conditions and has a low crystal tilt. Normal environmental conditions are defined as 20-25 ° C and up to 40% relative humidity. In the framework of the invention, the phrase "natural cannabinoid compounds, including unnatural derivatives of 10 cannabinoids, which derivatives can be obtained by derivatization of natural cannabinoids and are as unstable as natural cannabinoids. In the framework of the invention, the wording sugars include polysaccharides and the wording alcohols include polysaccharide alcohols. Preferred sugars in the present invention are non-reducing sugars. Non-reducing sugars are sugars that do not or cannot form reactive aldehyde or ketone groups. Examples of non-reducing sugars 15 Fructus jelly and sugars such as inulin. Preferred non-reducing sugars for use in the present invention are fructans or fructan mixtures. Fructans are understood to mean low Glycans or polysaccharides. Fructans may have a polydisperse chain length distribution, and may contain straight or branched chains. Preferably, fructans mainly contain bonds, as in inulin 2 but daggers may also contain β_2 6-bonds, as in levans. Suitable fructans can be derived directly from natural sources but can also undergo modifications. Examples of modifications are known reactions that lead to lengthening or shortening of the chain length. Except natural polysaccharides Of Polysaccharides such as chain-shortened hydrolysates and chain-length-fractionated grading products are also suitable in the present invention. The hydrolysis reaction for obtaining fructans with reduced chain length can be performed enzymatically (for example, using internal Inulinase), chemically (for example, using an acid in water), physical (for example, using heat) or by using heterogeneous catalysis (for example, using an acid ion exchanger). Fractionation of fructans such as inulin 5 It can be achieved by crystallization at low temperature, separation by column chromatography, membrane filtration and selection of alcohol with alcohol. Other fructans, such as long-chain fructans, can, for example, be obtained by removing monosaccharides and disaccharides from them Fructans are obtained by crystallization. Fructans with an enzymatically elongated chain length can also be used as the fructans of the present invention. In addition, reduced fructans can be used, which are their 10 reducing end groups (usually fructosyl Group) has been reduced, such as using sodium borohydride or hydrogen in the presence of a transition metal catalyst. Chemically modified fructans can also be used, Such as cross-linked fructans and hydroxyalkylated fructans. In all these fructans the average chain length is expressed as the number average degree of polymerization (DP). The abbreviation DP is defined as being in the oligomer or polymer 15 The average number of sugar units. The even more preferred reducing sugar in the present invention is inulin or an inulin mixture. Inulin is composed of β-1,2 bound fructose units and aDn at the reducing end of the molecule. Glycans or polysaccharides, and can have different degrees of polymerization (DP). The preferred inulin is an inulin with a DP greater than 6 or an inulin mixture with a DP greater than 6 for each of the inulins. Even more preferably the DP is 10 -30 inulin or inulin mixture. Most preferably inulin or inulin mixture with DP of 15-25. Inulin is particularly found in the roots and tubers of Liliaceae and Asteraceae. The most important source of inulin production is Jerusalem artichoke, dahlia and chicory root. Industrial production mainly uses chicory root as raw material. Derived from different natural sources 9 200403078 = The main difference between the powders is the degree of polymerization (Dp), which can be different. It is about 6 in the ginger and 1 in the chicory root.. Lv e is not in Dahlia to. Lik oligosaccharide or polysaccharide has favorable physicochemical properties when in the amorphous state, and can be used as an auxiliary substance in pharmaceutical preparations. These irrespective chemical properties are: (adjustable) high glass transition temperature, no reduction roadbed and usually low crystallinity. In addition, inulin is non-toxic and inexpensive. The weight of natural cannabinoid compounds on sugars or sugar alcohols W is more superior, and most preferred is Xin. ίο 15 20 The pharmaceutical composition of the present invention can be further processed into tablets such as ordinary oral tablets, sublingual tablets, lozenges, or orally disintegrated or dissolved tablets, capsules, lozenges, enemas, suppositories, for use in Products for transdermal administration, powders for pulmonary administration, or rods or syrups for subcutaneous or intramuscular administration. These forms of administration are known in the art and a person skilled in the art will be able to defend the composition of the invention into the desired form of administration. Preferred formulations are those for oral or pulmonary administration. A suitable technique for preparing the sugar glass of the present invention is cold green. Other technologies such as reading money and supercritical drying can also be made. The first step in the preparation of sugar glass that attracts the tianmudin compound is to prepare a solution in which these two substances are dissolved. Because of the hydrophilic nature of sugars and the lipophilic nature of natural cannabinoid compounds, these compounds are difficult to dissolve in the same solvent. It has now been discovered that this problem can be solved with mystery materials. Water is a good solvent for sugar and sugar alcohol 'and various organic solvents such as alcohols are good solvents for natural domain compounds. Due to the good mixing of water and alcohol, it is possible that both substances will achieve a certain degree of dissolution at a specific water / alcohol 10 200403078 ratio. Therefore, the present invention also relates to a method for preparing a pharmaceutical composition. The pharmaceutical composition includes a glass of a natural cannabinoid compound and a sugar or a sugar mixture, wherein the natural cannabinoid compound is introduced into the sugar glass 5 as a single molecular encapsulant without forming. A guest-guest complex characterized by a) dissolving the natural cannabinoid compound in a water-soluble organic solvent and the sugar or sugar mixture in water; b) dissolving the dissolved cannabinoid compound and the dissolved sugar or sugar The mixture is mixed to obtain a sufficiently stable mixture; 10 c) The mixture is freeze-dried, spray-dried, vacuum-dried or supercritically dried. Organic solvents suitable for forming a stable mixture with sugar, water and natural cannabinoid compounds are water-miscible solvents such as dimethylsulfoxide (DMSO), N, N-dimethylformamide (DMF), acetonitrile, acetic acid Ethyl esters and lower alcohols. Since the solvent must be removed by spray drying or freeze drying, the solvent should preferably also have a reasonable vapor pressure at the drying temperature. Lower alcohols are therefore preferred, defined as cardio-ditol, where the alkyl chain can be branched or straight. More preferred alcohols are c2-c4 alcohols such as ethanol, n-propanol and tert-butanol. The most preferred solvent is tert-butanol. 20 The ratio between the cannabinoid compound, solvent, water and sugar or sugar mixture should be chosen to obtain a sufficiently stable solution. A surfactant may be optionally added to improve stability. If no turbidity appears in the solution within the processing time, such as 120 minutes, 60 minutes, 30 minutes, or 10 minutes, the solution is judged to be sufficiently stable. For the spray drying process, the typical processing time is 30 minutes. For freeze-drying processes, the solution should be clear until frozen. The typical processing time here is 10 minutes. The water content after the drying process is preferably below 3%. The solvent content is preferably less than 3%. It is clear to those skilled in the art that the time required for drying can be derived from parameters such as sample thickness, sample temperature, pressure, and condenser temperature. Although the sugar stability of the cannabinoid compound was significantly improved by spray drying, the best results were obtained using a freeze-drying process. Therefore, the most preferred drying method in the present invention is freeze drying. 10 In the first stage of the freeze-drying process, the solution is frozen. This first stage should preferably be carried out quickly and the sample temperature should be lowered below Tg ', Ding 8' is the temperature of the Lengkang concentrated fraction (see DL Teagarden, Eur. J. Pharm. Sci., 15, 115-133 , 2002). Freeze drying below Tg 'produces porous cakes, while above Tg' results in collapsed cakes. Since a 15-hole porous cake can be more easily processed into a formulation such as a powder for tableting or pulmonary delivery, a porous cake is preferred. In addition, freeze-drying above Tg 'can cause sugar to crystallize. This prevents the drug from being introduced into the glass, with the result that the stabilizing effect is reduced. Brief description of the figure 20 Figure 1 shows the glass transition temperature of inulin under different humidity. Figure 2 is the water absorption isotherm of lyophilized inulin. Figure 3 shows the stabilization of a product freeze-dried from a water-TBA solution containing 4.00% THC.
第4圖係從水-1-丙醇溶液喷霧乾燥的，包含3.34%THC 12 200403078 的產物的穩定。 第圖係從水丙醇溶液噴霧乾燥的，包含7.77%THC 的產物的穩定。 第6圖係彳欠水-乙醇溶液喷霧乾燥的，包含4.00%THC的 5產物的穩定。 第7圖係物理混合物的降解。 第8圖係純THC的降解。 以下貝砭例僅旨在進一步更詳細地說明本發明，因 此此κ靶例不被視爲以任何方式限制本發明的範圍。 實施例1· Δ、四氫大麻酚菊粉玻璃的製備和性能· 材料 網乾類型 TEX!803 ’ 由荷蘭 Sensus，Roosendaal提 、、、屯化的Δ -四氫大麻紛（thc)是Unimed贈送的禮物。 所有其他化學品均是試劑級或分析級的，購自供應商。 方法 對菊粉進行物理化學表徵 菊粉聚合度的測定 菊粉的平均聚合度(DP)測定如下：通過加入3N HC1將 2〇菊粉溶液酸化至pH爲1.45。隨後，將溫度升高到80°C，通 過此操作菊粉降解成果糖和葡萄糖。在冷卻到室溫之後， 通過加入1.5 M NaOH將pH調節到6-8。通過HPLC確定果 糖/葡萄糖比例。使用AminexHPX-87C柱。以〇.6mL/min的 流速採用8(TC的MilliQ-水洗脫樣品。使用IR探測器測量果 13 糖和葡萄糖的量。DP是果糖含量和葡萄糖含量加一之比。 還原基團數目的測定 通過Sumner-測定根據以下步驟測定還原基團的數 目。製備20g酒石酸_NaK四水合物，lg二硝基水揚酸，lg NaOH和200mg苯酚在lOOmL水中的溶液。向l.5ml此溶液 中’加入l.OmL含有待分析糖的水溶液。隨後，向該混合 物中加入100从新鮮製備的〇.24MNa2S〇3水溶液。將獲得 的混合物渦旋混勻然後放入95°C的水浴中。15min之後， 將樣品從水浴取出，冷卻到室溫。在62〇ηιη下測量樣品的 /肖光。使用葡萄糖濃度爲0.10-1 .OOrng/mL的水溶液製備校 正曲線。測量進行一式三份。 差示掃描量熱法(DSC) 使用調製DSC(DSC 2920差示掃描量熱計，TA instruments，Gent，比利時）測量在 0%，45%和 60%RH下平 衡的凍幹菊粉的玻璃化轉變溫度（Tg)。採用每60秒士 0.318°C的調節幅度和2°C/min的加熱速率。在測量期間， 採用氮氣以35 mL/min的流速吹掃樣品池。取反向熱流對 溫度曲線中偏轉的中點作爲Tg。測量Tg—式兩份。 使用常規DSC測量60/40 v/v水/叔丁醇混合物中的 9.6%w/v菊粉溶液的冷凍濃縮級分的玻璃化轉變溫度 (Tg’）。以10〇C/min的冷卻速率將溶液冷卻到_7〇〇C 〇隨後， 採用2 °C /min的速率將樣品加熱到40 °C。在這些測量期 間，採用氦氣以35 mL/min的流速吹掃樣品池。取熱流對 溫度曲線中偏轉的中點作爲Tgf。測量Tg，一式兩份。 200403078 無定形菊粉的物理穩定性 爲評價無定形菊粉的物理穩定性，通過轉移到分別在 45%或60% RH下調節的氣候室，將通過冷凍乾燥獲得的無 定形菊粉的多孔餅狀物在20。(：下濕化。在平衡之後，目測 5判斷樣品，確定它們是否保持不變或出現崩塌。 動態蒸氣吸著 在環境壓力和25 °C下使用重量吸附分析儀(DVS_丨000 吸水设備 ’ Surface Measurement Systems Limited，倫敦， υκ)測罝凍幹菊粉的吸水等溫線。以1〇% RH爲階從〇%到 10 90% RH測量菊粉對水的吸收。最初的樣品重量爲約 10mg。在十分鐘時間内重量變化小於〇 9吨時，假定達到 平衡。 對THC進行物理化學表徵 在水中的溶解度 15 將純水加入過量1110中。使用磁力攪拌器在20°C下攪 拌所得的分散體。3天之後，對分散體進行離心並在21〇誰 波長下通過分光光度分析測定上清液中THC的濃度。採用 醇稀釋樣^口。使用已知濃度（丨·244-12·44 pg/mL)的THC：乙 醇溶液建立校正曲線。 20 動態蒸氣吸著 根據以上對於菊粉描述的程式測定THC的吸水率。將 TOC溶於f醇，然後放人㈣-讓伽巾。在初始曝露於 幹氮氣流過程中，甲醇蒸發。一旦約9〇%的溶劑蒸發，就 再將領外的THC溶液加入樣品杯中。重復此過程直到樣品 15 200403078 杯中存在15mg純THC。在最後的曱醇蒸發之後，以1〇%爲 階將相對濕度從0%增加到9〇〇/Q。 差示掃描量熱法(DSC) 用mDSC測定THC的熱行爲。使用每6〇秒土〇 318它的 5調節幅度和2°c/min的加熱速率。在測量期間，採用氮氣 以35 mL/min的流速吹掃樣品池。將—滴純THC放入樣品 杯中。在最初的冷卻之後對樣品進行第—次掃描直到5〇 °C。以此方式液滴能夠鋪展到樣品盤的整個底部，從而增 加在第二次掃描期間可用於傳熱的表面。然後將樣品冷卻 10 直到-40°C並加熱到350。(：。 包含THC的樣品的生産 製傷縣進行«軸或冷綠燥的溶液 15 20 製備三種不同的製劑用於喷霧乾燥以及一種製劑用 於冷康乾燥（表2)。分㈣過在水巾轉㈣和在 醇中溶解THC而製備製劑5,6,9和12。 田的 採用不同比例的水/妒、S、ηϊ m … ㈣過測試1G%_⑽溶液的 -性而研綱的合適體積比。將菊粉溶於不同量^ ㈣料後加人不同量的醇直_ 里^ THC遵循相同的程式，但將水加入th 對於 在加工時間内不出現萃濁，則溶液判斷爲果 喷霧乾燥，製備要求至多半小時嘴霧的批次物::此:於 至少在該時_ «是如的。對於冷 =此錢 應當是澄清的直到凍結，在這 "溶液 此外，研究了菊粉水溶下十分鐘就足夠了。 夜疋否可从緩慢加入或應當 16 200403078 合0 表2:用於^慶燥和冷凍气―燥的製劑 製劑 乾燥方法 溶劑 [菊粉] (mg/mL) THC/菊粉 (m%) 9 噴霧乾燥 H2O/EtOH=50/50(v/v) 47.73 4.00% 5 喷霧乾燥 H2O/l-PrOH=60/40(v/v) 49.00 3.34% 6 喷霧乾燥 H20/1 -PrOH=60/40(v/v) 46.17 7.77% 12 冷;東乾燥 H2O/t-BuOH=60/40(v/v 厂 96.00 4.00% 喷霧乾燥 使用michi 19〇微型喷霧乾燥器（Mchi，Flawil， 5 Switzerland)進行喷霧乾燥。典型的操作條件是根據如下設 疋值·鼠氣入口溫度· 148 C，得到出口溫度爲87，乾 燥氣流525 L/h ’吸氣器流量設定：2〇，和泵控制設定： 6。在喷務乾燥之後，將形成的粉末收集到⑽㈤^瓶子中， 採用氮氣吹洗約15分鐘。將産物在下貯存。 10 冷凍乾燥 使用 Christ model Alpha 2-4 凍幹機（Salm en Kipp， Breukelen，荷蘭）進行冷凍乾燥。在典型的實驗中，向 20mL小玻璃瓶中加入2_5mL溶液。將溶液在液氮中冷束， 隨後在-30 C的保存溫度、-53 °C的冷凝器溫度和〇.22〇 15 mBar的壓力下進行凍幹卜3天。隨後，在6小時内將保存溫 度逐漸升高到20°C，將壓力逐漸減低到0 〇5mBar。將樣品 置於真空乾燥器中貯存至少一天。 含THC的樣品的穩定性研究 在表3所示的五種不同條件下貯存樣品。以不同的時 20間間隔取出樣品，通過HPLC測定未降解的THC的量。將 17 200403078 純THC和THC與菊粉的物理混合物用作對照。純THC樣品 製備如下。將720.5mg的THC溶於20.00mL甲醇。將70μί此 溶液轉移到直徑爲24mm的玻璃小瓶中。隨後使溶劑在幹氮 氣流中蒸發，在小瓶中留下2.52mg的純THC。通過稱量約 5 192mg的菊粉放入直徑24mm的小瓶中而製備物理混合物。 隨後，加入200 pL 36.025 mg/mL的THC甲醇溶液，得到包 含按質量計4.0% THC的混合物。 表3:含THC的樣品的貯存條件 溫度ΓΟ 相對濕度（％) 氣氛 20 0 低[〇2] 20 45 空氣 20 60 空氣 47 0 低[〇2] 47 「5 空氣 10 THC·分析 通過HPLC分析樣品。製備如下：將曱醇加入樣品 中。十分鐘的超聲處理使産物分散在甲醇中。用手搖動如 此獲得的懸浮液。在提取兩天之後，取出樣品。將樣品離 心並將上清液用曱醇稀釋。在對照實驗中，顯示超聲處理 15未誘導THC降解。在兩天提取期間，沒有測量到THc的顯 著降解。使用裝配有光二極體陣列紫外—可見光檢測器 (Shimadzu SPD-M6A型號)和 Chr〇mpack Ν— 1〇〇 ⑽ 柱㈣心麵）的ISCO 235〇型號系統。採用κ〇_η InStmmentS HPLC細自動取樣器注射樣品（2〇吣），採用 18 200403078 甲醇/水=86/14 (v/v)的混合物洗脫。流速是1.5 mL/min。在 214nm下測量吸光度。使用SPD-MXA軟體分析收集到的資 料。在未經處理的THC的色譜中，在7.5min保留時間觀察 到一個大峰。在有意地部分降解的THC的色譜中，在 5 7.5min保留時間的峰變小而在更短的保留時間出現新峰。 在7.5 min保留時間的峰歸於A9-THC。其他峰歸於降解産 物。從在7.5min洗脫時間的峰下的面積計算加工樣品中（未 降解的）THC的含量。使用已知濃度（0-122 pg/mL)的THC 甲醇溶液建立校正曲線。在每一次HPLC-運行中，包括一 10 些校正點。4°c下在2周時間内用於此目的的溶液未顯示出 顯著的降解。測量至少進行一式兩份。 結果 對菊粉進行物理化學表徵 所使用的菊粉的物理化學特性見表4。 15 表4:菊粉玻璃的物理化學表徵 平均聚合度 23 包含還原基圑的糖單元％ 5.9±0_1 Tg 155_4 士 0.1 〇C Tg， -24〇C 在2(TC下的物理穩定性 在RHS45%時穩定；在RH260%時崩塌 吸濕性 質量變化=0.22*RH(%)+0.61 發現菊粉的DP爲23。出於幾種原因此數值應當被認 爲是指示值。菊粉由末端爲(1-0-(1—2户比喃型葡萄糖環的 直鏈P-D-(2—l)鍵合的果糖低聚物組成。因此，可以從在 20 此呈現的葡萄糖/果糖比例計算DP。然而，市售菊粉可能 19 200403078 包含葡萄糖端基被裂解的菊粉種類。這些種類的存在會 =起對DP估叶過高。另一方面市售菊粉也可能包含少量 葡萄糖。这些種類的存在會引起對DP的低估。 由於在單糖環之間的特定鍵合，菊粉應當不包含還 5，基團。然而，S_er測定顯示用於此研究的菊粉的糖 皁7〇_5.9±〇.1%包含還原基團。還原基團的存在可能主 要歸於其葡萄糖端基被裂解的菊粉種類，儘管單糖的存 在也可能在其中起作用。這些單糖可以是葡萄糖和果糖。 果糖是非還原糖。然而，在Su_r測定期間，糖經受高 1〇溫，由此果糖可容易地變爲葡萄糖(Lobry de Bruyn van Ekenstein重排）。f際上在對照實驗中，人們發現果糖在 測定十顯示每個分子一個還原基團（資料未顯示）。因此， 還原基團的實測量可能被高估了。 發現菊粉的玻璃化轉變溫度（Tg). 155 4±〇 rc。此 15數值大大高於如下物質的Tg :海藻糖（120。〇和蔗糖（76 t)，這兩種糖通常用於使不穩定的藥物變得穩定。由於 在高於Tg的溫度下物質變成橡膠態，因此高丁§是重要 的。與玻螭態相比，在橡膠態中分子流動性強烈增加， 結果使得包封入的藥物物質的降解速率強烈增加。除了 20該情況以外，在橡膠態中還可發生結晶。在結晶過程中， 引入的藥物物質從穩定基質中排出而完全喪失保護。 可以似乎非常高。然而，在曝露於濕潤空氣時糖破璃吸 水（見下文）。水起到的作用是作爲糖玻璃的增塑劑並強別 地降低Tg。因此，在Tg降低到室溫之前，菊粉玻璃可比 20 200403078 海藻糖或蔗糖玻璃吸收多得多的水。 發現菊粉的Tg’爲-24°C。同樣此數值高於如下物質的 Tg’ ：海藻糖(-36t)和蔗糖(-39°C)。當選擇冷凍乾燥作爲 乾燥方法時，優選Tg’相對較高，這是因爲樣品溫度應當 5 保持低於Tg’。當樣品溫度高於Tg’時，冷凍濃縮級分處於 橡膠態，如上所述分子流動性相對較高。由於藥物物質 在冷凍濃縮級分中的濃度非常高，當與起始溶液相比較 時降解速率可增加。而且在此情況下可能還易於發生糖 的結晶並伴隨對藥物物質的劣化效應。此外在低於Tg’進 10 行冷凍乾燥産生多孔餅狀物，而高於Tg’則獲得崩塌的餅 狀物。由於可以更容易地加工成如用於壓片的粉末或肺 輸送製劑，因此優選多孔餅狀物。 通過將玻璃曝露於各種相對濕度的空氣而評價菊粉 玻璃在20°c的物理穩定性。我們發現由冷凍乾燥製備的 15 菊粉多孔餅狀物至多到45%的RH仍保持不受影響。然 而，在60%的RH下，多孔餅狀物發生崩塌。這意味著在 45%-60%的RH下，樣品吸收水的程度使得超越了 Tg。短 時間曝露於60%RH可以應用於凍幹餅以使它部分崩塌。 此部分崩塌的物質可形成具有足夠強度的合適速溶片 20 劑。在0，45%和60%RH中平衡之後凍幹菊粉的Tg見第1 圖。 使用重量吸著分析儀測量在25°C下曝露於相對濕度爲 0-90%的空氣的凍幹菊粉的水分吸收。在整個範圍的相對 濕度内，發現在水分吸收與樣品曝露的RH之間存在線性 21 200403078 關係（表5 ;第2圖）。如以上所發現的那樣，在45%_6〇%的 RH下跨越了 Tg。線性關係指示在實驗的時限(小時)期間， 菊粉沒有發生結晶。當發生結晶並形成無水晶體時，樣品 的含水量會下降到接近於零。另一方面當形成包入水分^ 5的晶體時，樣品的含水量隨RH增加而保持或多或少基本 相同。對非晶形糖如海藻糖、蔗糖和乳糖進行吸水試驗時 觀察到這些現象。因此，結果指示無定形菊粉不如非晶形 海藻糖、蔗糖和乳糖那樣容易結晶。 對THC進行物理化學表徵 G 溶解度 發現THC的溶解度小於ipg/mL (大約〇·5㈣眺）。 動態蒸氣吸著 在曝露於90%RH之後發現純THC僅吸收〇·3%水。此 吸水程度可能歸因於水分被吸附到T H c上而不是吸收 15 THC 内。 差示掃描量熱法 在THC的差示熱分析圖中發現Tg爲1〇。〇。此外發現一 個在200°C開始的吸熱峰。從熱力學的觀點來看，可以預 20 功剛向於Tg就出現結晶。然而，衆所周知THC並不容易社 晶。因而’在環境溫度下，THC處於橡膠態或液態。 峰歸於蒸發。 …、 包含THC的樣品的生産 用於喷霧乾燥或冷凍乾燥的水-鏈燒醇溶液 將二種相關的醇加入菊粉水溶液中。測定所獲得的溶Figure 4 shows the stabilization of a product spray-dried from a water-1-propanol solution containing 3.34% THC 12 200403078. The figure shows the stabilization of a product containing 7.77% THC spray-dried from a water propanol solution. Figure 6 shows the stabilization of 5 products containing 4.00% THC, spray-dried with an aqueous-ethanol solution. Figure 7 is the degradation of the physical mixture. Figure 8 is the degradation of pure THC. The following examples are only intended to further illustrate the present invention in more detail, and thus the kappa target is not to be considered as limiting the scope of the present invention in any way. Example 1. Preparation and properties of Δ and tetrahydrocannabinol inulin glass. Material mesh dry type TEX! 803 ′ Δ-tetrahydrocannabinoid (thc) from Sensus, Roosendaal in the Netherlands was presented by Unimed. one's gift. All other chemicals are reagent grade or analytical grade and were purchased from suppliers. Methods Physical and chemical characterization of inulin was determined. The degree of polymerization of inulin was determined as follows: The 20 degree inulin solution was acidified to pH 1.45 by adding 3N HC1. Subsequently, the temperature was raised to 80 ° C, and inulin was used to degrade fructose and glucose. After cooling to room temperature, the pH was adjusted to 6-8 by adding 1.5 M NaOH. The fructose / glucose ratio was determined by HPLC. Aminex HPX-87C column was used. Samples were eluted with 8 ° C MilliQ-water at a flow rate of 0.6 mL / min. The amount of fructose and glucose was measured using an IR detector. DP is the ratio of fructose content to glucose content plus one. Number of reducing groups Determination The number of reducing groups was determined by Sumner-assay according to the following procedure. A solution of 20 g of tartaric acid-NaK tetrahydrate, lg dinitrosalicylic acid, lg NaOH and 200 mg of phenol in 100 mL of water was prepared. 'Add 1.0 mL of an aqueous solution containing the sugar to be analyzed. Subsequently, add 100 freshly prepared 0.24 M Na2SO3 aqueous solution to the mixture. The obtained mixture is vortexed and then placed in a 95 ° C water bath. 15 min After that, the sample was taken out of the water bath and cooled to room temperature. The sample's / Xiao Guang was measured at 6200 nm. A calibration curve was prepared using an aqueous solution with a glucose concentration of 0.10-1.000 rng / mL. The measurement was performed in triplicate. Difference Scanning calorimetry (DSC) The glass transition temperature of lyophilized inulin balanced at 0%, 45%, and 60% RH was measured using a modulated DSC (DSC 2920 differential scanning calorimeter, TA instruments, Gent, Belgium). (Tg). Use every 60 The adjustment range of ± 0.318 ° C and the heating rate of 2 ° C / min. During the measurement, the sample cell was purged with nitrogen at a flow rate of 35 mL / min. The midpoint of the deflection in the reverse heat flow versus temperature curve was taken as the Tg. Measure Tg in duplicate. Measure glass transition temperature (Tg ') of freeze-concentrated fractions of 9.6% w / v inulin solution in 60/40 v / v water / tert-butanol mixture using conventional DSC. Take 10 The cooling rate of 〇C / min cooled the solution to -700C. 〇 Subsequently, the sample was heated to 40 ° C at a rate of 2 ° C / min. During these measurements, helium was used at a flow rate of 35 mL / min. Purge the sample cell. Take the midpoint of the deflection in the heat flow versus temperature curve as Tgf. Measure Tg in duplicate. 200403078 Physical stability of amorphous inulin To evaluate the physical stability of amorphous inulin, transfer to In a climate chamber adjusted at 45% or 60% RH, the porous cakes of amorphous inulin obtained by freeze-drying are wetted at 20 ° (: wet down. After equilibration, visually inspect the samples to determine whether they remain Change or collapse. Dynamic vapor sorption at ambient pressure and 25 ° C A gravimetric adsorption analyzer (DVS_ 丨 000 water absorption device 'Surface Measurement Systems Limited, London, υκ) was used to measure the water absorption isotherm of lyophilized inulin. From 10% to 10 90% RH in steps of 10% RH. The absorption of water by inulin was measured. The initial sample weight was about 10 mg. When the weight change is less than 0.9 tons in ten minutes, equilibrium is assumed to be reached. Physical and chemical characterization of THC Solubility in water 15 Add pure water to an excess of 1110. The resulting dispersion was stirred using a magnetic stirrer at 20 ° C. After 3 days, the dispersion was centrifuged and the concentration of THC in the supernatant was determined by spectrophotometric analysis at a wavelength of 210 nm. Samples were diluted with alcohol. A calibration curve was established using a known concentration (丨 · 244-12 · 44 pg / mL) of THC: ethanol. 20 Dynamic Vapor Sorption The water absorption of THC was determined according to the procedure described above for inulin. TOC was dissolved in f-alcohol, and then put in a cymbal-gray towel. During the initial exposure to a stream of dry nitrogen, methanol evaporated. Once about 90% of the solvent has evaporated, add extra THC solution to the sample cup. This process was repeated until 15 mg of pure THC was present in the sample 15 200403078 cup. After the final methanol was evaporated, the relative humidity was increased from 0% to 900 / Q in 10% steps. Differential Scanning Calorimetry (DSC) mDSC was used to determine the thermal behavior of THC. It was adjusted to 5 times every 60 seconds and adjusted at a temperature of 2 ° C / min. During the measurement, the sample cell was purged with nitrogen at a flow rate of 35 mL / min. Put a drop of pure THC into the sample cup. Scan the sample for the first time after initial cooling until 50 ° C. In this way the droplets can spread over the entire bottom of the sample pan, increasing the surface available for heat transfer during the second scan. The sample was then cooled for 10 to -40 ° C and heated to 350. (:. The production of samples containing THC was performed in the injured county «shaft or cold green dry solution 15 20 Three different preparations were prepared for spray drying and one preparation was used for cold drying (Table 2). Preparations 5, 6, 9 and 12 were prepared by converting water towels and dissolving THC in alcohol. The field was tested with different ratios of water / jealousy, S, ηϊ m… Appropriate volume ratio. Dissolve inulin in different amounts and add different amounts of alcohol. THC follows the same procedure, but add water to th. For turbidity that does not occur during processing time, the solution is judged as If spray-dried, prepare batches that require up to half an hour of mouth spray :: this: at least at that time _ «as it is. For cold = this money should be clarified until frozen, in this " solution In addition, It has been studied that inulin dissolves in water for ten minutes is sufficient. Is it possible to add slowly at night or should be 16 200403078 0 0 Table 2: Preparations for ^ Qingzao and frozen air-drying preparations Formulation drying method Solvent [inulin] (mg / mL) THC / inulin (m%) 9 spray-dried H2O / EtOH = 50/50 (v / v) 47.73 4.00% 5 spray-dried H 2O / l-PrOH = 60/40 (v / v) 49.00 3.34% 6 Spray drying H20 / 1 -PrOH = 60/40 (v / v) 46.17 7.77% 12 Cold; East drying H2O / t-BuOH = 60 / 40 (v / v factory 96.00 4.00% spray drying) Use the michi 19〇 micro spray dryer (Mchi, Flavil, 5 Switzerland) for spray drying. Typical operating conditions are set according to the following settings: • Rat gas inlet temperature · 148 C, get an outlet temperature of 87, a dry air flow of 525 L / h 'aspirator flow setting: 20, and pump control setting: 6. After spray drying, collect the formed powder into a ⑽㈤ ^ bottle, Purge with nitrogen for about 15 minutes. Store the product underneath. 10 Freeze-drying Freeze-drying using a Christ model Alpha 2-4 lyophilizer (Salm en Kipp, Breukelen, The Netherlands). In a typical experiment, a 20 mL glass vial is used. 2-5 mL of solution was added to the solution. The solution was cooled in liquid nitrogen, and then lyophilized for 3 days at a storage temperature of -30 C, a condenser temperature of -53 ° C, and a pressure of 0.220 15 mBar. Subsequently, Gradually increase the storage temperature to 20 ° C and gradually reduce the pressure to 0.05 mBar within 6 hours. Storing at least one day in a desiccator. Stability of THC containing samples are stored under five different conditions shown in Table 3 samples. At different time intervals samples were taken 20, the amount of undegraded THC was determined by HPLC. 17 200403078 Pure THC and a physical mixture of THC and inulin were used as controls. Pure THC samples were prepared as follows. 720.5 mg of THC was dissolved in 20.00 mL of methanol. 70 μL of this solution was transferred to a glass vial with a diameter of 24 mm. The solvent was then evaporated under a stream of dry nitrogen, leaving 2.52 mg of pure THC in the vial. A physical mixture was prepared by weighing about 5 192 mg of inulin into a 24 mm diameter vial. Subsequently, 200 pL of 36.025 mg / mL of a THC methanol solution was added to obtain a mixture containing 4.0% THC by mass. Table 3: Storage conditions for THC-containing samples Temperature ΓΟ Relative humidity (%) Atmosphere 20 0 Low [〇2] 20 45 Air 20 60 Air 47 0 Low [〇2] 47 「5 Air 10 THC · Analysis Sample by HPLC Prepared as follows: methanol is added to the sample. Ten minutes of sonication disperses the product in methanol. The suspension so obtained is shaken by hand. After two days of extraction, the sample is removed. The sample is centrifuged and the supernatant is used. Alcohol dilution. In a control experiment, it was shown that sonication 15 did not induce THC degradation. During two days of extraction, no significant degradation of THc was measured. A UV-visible light detector (Shimadzu SPD-M6A model equipped with a photodiode array was used) ) And Chrompack N—100 (column core)) ISCO 235 model system. Samples were injected using a κ〇_η InStmmentS HPLC fine autosampler (20), using 18 200403078 methanol / water = 86 / 14 (v / v) mixture was eluted. The flow rate was 1.5 mL / min. The absorbance was measured at 214 nm. The collected data was analyzed using SPD-MXA software. In untreated THC chromatography, retained at 7.5 min View of time A large peak was observed. In the chromatogram of THC that was intentionally partially degraded, the peak at a retention time of 5 7.5 min became smaller and a new peak appeared at a shorter retention time. The peak at a retention time of 7.5 min was attributed to A9-THC. Other The peak is attributed to degradation products. The content of (undegraded) THC in the processed sample was calculated from the area under the peak at 7.5 min elution time. A calibration curve was established using a THC methanol solution of known concentration (0-122 pg / mL). In each HPLC-run, a few 10 calibration points were included. The solution used for this purpose at 4 ° C over 2 weeks did not show significant degradation. Measurements were performed in at least duplicates. Results Physical properties of inulin The physical and chemical characteristics of inulin used for chemical characterization are shown in Table 4. 15 Table 4: Physical and chemical characterization of inulin glass Average polymerization degree 23 Sugar unit containing reduced hydrazone% 5.9 ± 0_1 Tg 155_4 ± 0.1 〇C Tg,- The physical stability at 24 ° C at 2 ° C is stable at 45% RHS; the collapse hygroscopic mass change at RH260% = 0.22 * RH (%) + 0.61 DP of inulin was found to be 23. For several reasons This value should be considered as an indicator. It is a composition of (1-0- (1-2) pyranose-type glucose ring with a linear PD- (2-1) bonded fructose oligomer. Therefore, DP can be calculated from the glucose / fructose ratio presented at 20. However, commercially available inulin may 19 200403078 contain inulin species whose glucose end groups are cleaved. The existence of these species will result in an overestimate of DP. On the other hand, commercially available inulin may also contain a small amount of glucose. The presence of these species can cause an underestimation of DP. Due to the specific bonding between the monosaccharide rings, inulin should not contain any further 5, groups. However, the S_er assay showed that 70% to 5.9 ± 0.1% of the inulin sugar soap used in this study contained reducing groups. The presence of reducing groups may be mainly due to the inulin species whose glucose end groups are cleaved, although the presence of monosaccharides may also play a role here. These monosaccharides can be glucose and fructose. Fructose is a non-reducing sugar. However, during the Su_r assay, sugar was subjected to a high temperature of 10, whereby fructose could easily be changed to glucose (Lobry de Bruyn van Ekenstein rearrangement). In the control experiments, it was found that fructose showed ten reducing groups per molecule (data not shown). Therefore, the actual measurement of the reducing group may be overestimated. The glass transition temperature (Tg) of inulin was found to be 155 4 ± 〇 rc. This 15 value is much higher than the Tg of trehalose (120.0) and sucrose (76 t). These two sugars are commonly used to stabilize unstable drugs. Because the substance becomes at a temperature above Tg The rubbery state, therefore, Gao Ding § is important. Compared with the glassy state, the molecular mobility is strongly increased in the rubbery state, resulting in a strong increase in the degradation rate of the encapsulated drug substance. Except for 20 cases, in the rubbery state Crystallization can also occur in the process. During the crystallization process, the introduced drug substance is discharged from the stable matrix and completely loses its protection. It can seem very high. However, sugar breaks glass and absorbs water when exposed to humid air (see below). The role is to act as a plasticizer for sugar glass and strongly reduce Tg. Therefore, inulin glass can absorb much more water than trehalose or sucrose glass before 20 200403078. Tg of inulin is found 'It is -24 ° C. This value is also higher than the Tg of the following: trehalose (-36t) and sucrose (-39 ° C). When freeze drying is selected as the drying method, Tg' is preferably relatively high, which Because of The product temperature should be kept below Tg '. When the sample temperature is higher than Tg', the freeze-concentrated fraction is in a rubbery state, as described above, the molecular mobility is relatively high. Because the concentration of the drug substance in the freeze-concentrated fraction is very high The degradation rate can be increased when compared with the starting solution. In this case, the crystallization of sugar may be prone to occur along with the degradation effect on the drug substance. In addition, freeze-drying at 10 below Tg 'produces a porous cake. Above Tg ', a collapsed cake is obtained. Porous cakes are preferred because they can be more easily processed into powders or pulmonary delivery formulations such as those used for tabletting. Porous cakes are preferred by exposing glass to various relative humidity The physical stability of inulin glass at 20 ° C was evaluated by air. We found that up to 45% RH of 15 inulin porous cakes prepared by freeze-drying remained unaffected. However, at 60% RH, The porous cake collapsed. This means that at 45% -60% RH, the sample absorbed water so that it exceeded Tg. Exposure to 60% RH for a short time can be applied to freeze-dried cake to partially collapse it. This partially collapsed material can form a suitable instant tablet with sufficient strength. 20 Tg of lyophilized inulin after equilibration in 0, 45% and 60% RH is shown in Figure 1. Measured at 25 ° using a gravimetric analyzer Moisture absorption of lyophilized inulin exposed to air with a relative humidity of 0-90% at C. A linear 21 200403078 relationship was found between the moisture absorption and the RH of the sample exposure over the entire range of relative humidity (Table 5; (Figure 2). As found above, Tg was crossed at 45% to 60% RH. A linear relationship indicates that inulin did not crystallize during the experimental time period (hours). When crystallisation occurred and anhydrous crystals formed At this time, the water content of the sample will drop to near zero. On the other hand, when crystals containing water ^ 5 are formed, the water content of the sample remains more or less the same as RH increases. These phenomena were observed in water absorption tests on amorphous sugars such as trehalose, sucrose and lactose. Therefore, the results indicate that amorphous inulin is not as easy to crystallize as amorphous trehalose, sucrose, and lactose. Physical and chemical characterization of THC G Solubility The solubility of THC was found to be less than ipg / mL (approximately 0.5 °). Dynamic vapor sorption After exposure to 90% RH, it was found that pure THC absorbed only 0.3% water. This degree of water absorption may be due to water being adsorbed onto T H c instead of absorbing within 15 THC. Differential Scanning Calorimetry Tg was found to be 10 in the differential thermal analysis chart of THC. 〇. In addition, an endothermic peak was started at 200 ° C. From a thermodynamic point of view, it can be predicted that crystals appear as soon as the work is directed to Tg. However, it is well known that THC is not easily crystallized. Thus, 'THC is rubbery or liquid at ambient temperature. The peak is attributed to evaporation. …, Production of samples containing THC Water-chain alcohol solution for spray drying or freeze drying Two related alcohols are added to the aqueous inulin solution. Determine the obtained solvent
22 200403078 液^呆持澄清多長時間。在4mL水中_g㈣之後，加 和/或醇至總體積1〇mL，得到增液。這樣獲得 醉的最大濃度。將THC溶於感興趣的醇中。隨後，加入醇 ^水以得到G.4%W/V溶液。在表5中給出獲得穩定溶液 (在材料和方法中定義）所需的組成。22 200403078 How long does the liquid clarify. After 4 g of gadolinium in 4 mL of water, add and / or alcohol to a total volume of 10 mL to obtain a liquid enhancement. This gives the maximum concentration of drunk. THC is dissolved in the alcohol of interest. Subsequently, alcohol and water were added to obtain a G.4% W / V solution. The composition required to obtain a stable solution (defined in Materials and Methods) is given in Table 5.
ίοίο
通過將菊粉水溶液加人THC溶液中製備躲喷霧乾燥 的溶液。證明這必須相當快速地進行，叫止菊粉使混合 物渾濁。溶液在«溶液所㈣時間㈣持澄清。通過在 20mL TBA中溶解69〇mg THC製備要冷滚乾燥的thc溶 液。在每個20mL小玻璃管中注入〇.2SmLTHC溶液。隨 後，將溶液用0.57mL純TBA稀釋。然後，加入12mL菊粉 水溶液((160mg/mL)，用手搖動小瓶隨後立即冷凍。A spray-dried solution was prepared by adding an aqueous inulin solution to the THC solution. It turned out that this had to be done fairly quickly, and called inulin to make the mixture cloudy. The solution was kept clear for «solution time. A thc solution to be cold-roll dried was prepared by dissolving 690 mg of THC in 20 mL of TBA. 0.2SmL THC solution was injected into each 20 mL small glass tube. Subsequently, the solution was diluted with 0.57 mL of pure TBA. Then, 12 mL of an inulin aqueous solution ((160 mg / mL)) was added, and the vial was shaken by hand and immediately frozen.
在乾燥之後回收THC 在生産之後即刻在噴霧乾燥樣品中的THC量低於期待 值。發現初始回收率爲約5〇%。在將霧化氣流和來自加熱 為的氣流爲氮氣之後，回收率增加到75%。在冷凌乾燥 的情況下，發現在乾燥工藝之後的樣品中THC量爲1〇〇%預 期值。 23 200403078 對包含™c的樣品進行表徵 掃描電鏡 喷霧餘產物的掃描電鏡(SEM)照片顯示存在小顆粒 附聚物。這些顆粒直徑爲1-5 μηι，是中空的。喷霧乾燥顆 5粒的小尺寸和密度降低使得它們非常適於加工成吸入用乾 粉製劑。參考産物（無THC的菊粉，在相同條件下乾燥並 採用相同的溶劑）的SEM照片顯示沒有差異。在包含THC 的樣品的顆粒表面上沒有注意到THC斑點，指示THC被引 入菊粉基質中。 10 含THC的樣品的穩定性 分別在20°C和47t;下，將樣品曝露於含有〇2或低仏條 件（圖中指示爲氮氣）。此外，在2〇。〇下將它們曝露於兩種不 同的濕度，如之前所概括的那樣。在從喷霧乾燥器收集之 後，噴霧乾燥的産物顯示出輕微的顏色變化。 第3-6圖顯不批次物12，5，6和9的結果。測定THC的 夏。在圖令，將在幾次曝露時間之後樣品中存在的Δ9-ΤΗ(：： 分數對五種不同的氣候作圖。 凍幹樣品（批次物12)見第3圖。在前述五種氣候之 後，將此批次物的部分樣品曝露於60°C 〇%RH。第4圖顯 下攸Ι-PrOH和水的溶液喷霧乾燥的包含3·34% THC的批 人物的穩定性貧料，第5圖顯示具有更高THC含量7.77% 仁也疋從水-1-丙醇溶液喷霧乾燥的批次物。第6圖顯示從 乙醇和水的溶液噴霧乾燥的包含4.00% THC的批次物的 穩定性資料。 .1.4 24 200403078 從喷霧乾燥批次物的結果顯示，製劑改善了 THC的 穩定性。溫度對降解速率影響最大。水分和氧氣重要性 較低。然而，應當注意到在氮氣下貯存的樣品可能受到 某種程度的氧氣污染。 5 不同的圖清楚地顯示，當與物理混合物和純THC相 比時，冷凍乾燥産物的穩定性優異（參見第7和8圖）。顯然 製備糖玻璃的方法強烈地影響産物的穩定性。 如可以從第5圖中看出的那樣，對於所有測試條件， 除了 60% RH，冷凍乾燥産物中的降解最少。然而在此發 10 現的略爲低的濃度也可能是由如下事實引起的··在此條件 下物質崩塌，使提取過程有效性減低。 參考批次物 爲測試菊粉的穩定能力，應當將以上顯示的資料與 具有相同化學和物理結構但不含菊粉的批次物相比較。 15 這意味著參考批次物由單獨的菊粉分子，事實上THC蒸 氣組成。由於這是不切實際的，製備了兩種其他參考批 次物：包含約4% THC和96%未加工菊粉的物理混合物和 純THC。結果分另J見第7和8圖。 必須提及的是在物理混合物的製備期間，THC的曱醇 20 溶液將菊粉粉末軟化至一定程度。在甲醇蒸發之後，或多 或少的菊粉和THC的固體膜出現在小瓶底部。該低孔隙率 薄膜産生對此參考物質的額外保護。除此之外，可能THC 甲醇溶液與糖的混合已經導致一部分THC被包入。 應當強調在純THC樣品中自保護也是有關的，因爲它 25 200403078 們也形成防護膜。 【圖式簡單說明3 第1圖係在不同濕度下菊粉的玻璃化轉變溫度。 第2圖係凍干菊粉的吸水等溫線。 5 第3圖係從水-TBA溶液冷凍乾燥的，包含4.00%THC的 產物的穩定。 第4圖係從水-1-丙醇溶液喷霧乾燥的，包含3.34%THC 的產物的穩定。 第5圖係從水-1-丙醇溶液喷霧乾燥的，包含7.77%THC 10 的產物的穩定。 第6圖係從水-乙醇溶液喷霧乾燥的，包含4.00%THC的 產物的穩定。 第7圖係物理混合物的降解。 第8圖係純THC的降解。 15 【圖式之主要元件代表符號表】 無 26THC recovered after drying The amount of THC in the spray-dried sample immediately after production was lower than expected. The initial recovery was found to be about 50%. After the atomizing gas stream and the heating gas stream were nitrogen, the recovery was increased to 75%. In the case of chilling and drying, it was found that the amount of THC in the sample after the drying process was 100% of the expected value. 23 200403078 Characterization of samples containing ™ c. Scanning electron microscopy. Scanning electron microscopy (SEM) photographs of the spray residue show small particle agglomerates. These particles are 1-5 μηι in diameter and are hollow. The small size and reduced density of spray-dried granules makes them very suitable for processing into dry powder formulations for inhalation. SEM photos of the reference product (THC-free inulin, dried under the same conditions and using the same solvent) showed no difference. No THC spots were noticed on the particle surface of the THC-containing sample, indicating that THC was introduced into the inulin matrix. 10 Stability of THC-containing samples at 20 ° C and 47t, respectively. Expose the samples to conditions containing 0 2 or low radon (indicated as nitrogen in the figure). In addition, at 20. They were exposed to two different humidity levels, as outlined earlier. After collecting from the spray dryer, the spray-dried product showed a slight color change. Figures 3-6 show the results for batches 12, 5, 6, and 9. The summer of THC was measured. In the drawing, the Δ9-ΤΗ (:: fractions present in the sample after several exposure times are plotted against five different climates. The lyophilized sample (batch 12) is shown in Figure 3. In the aforementioned five climates Afterwards, a portion of this batch of samples was exposed to 60 ° C 0% RH. Figure 4 shows the stability of the batches containing 3.34% THC in a spray-dried solution of I-PrOH and water. Figure 5 shows a batch with a higher THC content of 7.77%. Renye also spray-dried a batch from a water-1-propanol solution. Figure 6 shows a spray-dried batch containing 4.00% THC from a solution of ethanol and water. Material stability data. 1.4 24 200403078 Results from spray-dried batches show that the formulation improves THC stability. Temperature has the greatest effect on the rate of degradation. Moisture and oxygen are less important. However, it should be noted that Samples stored under nitrogen may be contaminated to some degree by oxygen. 5 The different plots clearly show that the stability of the freeze-dried product is excellent when compared to physical mixtures and pure THC (see Figures 7 and 8). Obviously The method of preparing sugar glass strongly affects the stability of the product Qualitative. As can be seen in Figure 5, for all test conditions, except for 60% RH, the degradation in the freeze-dried product is minimal. However, the slightly lower concentration found here may also be due to the following facts Caused by the material collapse under this condition, reducing the effectiveness of the extraction process. The reference batch is to test the stability of inulin. The data shown above should be compared with the batch with the same chemical and physical structure but without inulin. This compares with the second batch. 15 This means that the reference batch consists of separate inulin molecules, in fact THC vapor. Since this is impractical, two other reference batches were prepared: containing about 4% THC and The physical mixture of 96% raw inulin and pure THC. The results are shown separately in Figures 7 and 8. It must be mentioned that during the preparation of the physical mixture, the THC 20 alcohol solution softens the inulin powder to a certain degree A solid film of more or less inulin and THC appears at the bottom of the vial after the methanol has evaporated. This low porosity film creates additional protection for this reference substance. In addition, THC methanol solutions and sugars may be Mixing has already caused a part of THC to be encased. It should be emphasized that self-protection is also relevant in pure THC samples, as it also forms a protective film. [Schematic description 3 Figure 1 is the inulin in different humidity Glass transition temperature. Figure 2 is the water absorption isotherm of lyophilized inulin. 5 Figure 3 is the stability of the product freeze-dried from water-TBA solution and contains 4.00% THC. Figure 4 is from water-1 -Stability of spray-dried propanol solution containing 3.34% THC. Figure 5 shows the stabilization of product spray-dried from water-1-propanol solution containing 7.77% THC 10. Figure 6 is the stabilization of a product spray-dried from a water-ethanol solution containing 4.00% THC. Figure 7 is the degradation of the physical mixture. Figure 8 is the degradation of pure THC. 15 [Representation of the main components of the diagram] None 26