TW201416074A - Analgesic (sebacoyl dinalbuphine ester) PLGA controlled release dosage form - Google Patents

Abstract

The present invention features a long term controlled release formulation of the nalbuphine pro-soft drug, Sebacoyl dinalbuphine ester, in combination with commonly used pharmaceutical excipient biodegradable polymer PLGA. Said formulaion was selected from the following groups of pharmaceutical formulations including such as: tablets, capsules, soft capsules, granules, suspensions, microspheres, oral implants, implantable injections and others. Said long term controlled release formulation significantly improved the dosage and frequency for administering nalbuphine to once per half month or few months, compared to four to six times per day in the traditional way, which is one of the major features and effects of the present invention. The major improvement of this inventioncan be achieved by confirmation of the pharmacokinetic profiles and the duration time of efficacy level of drug through in vivo experiments, subsequently improved the dosage and frequency of the traditionally used nalbuphine injections.

Description

長效止痛藥癸二醯雙那布扶林酯-PLGA控釋劑型 Long-acting analgesic drug 癸二醯双那布福林酯-PLGA controlled release dosage form

本發明係關於用以治療哺乳動物急性或慢性疼痛之化合物、前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)、與藥學常用生物可分解高分子PLGA賦型劑之組成物，並將該組合物製備成可進入人體之長效控釋劑型。藉由高分子的而達到長效控制藥物釋放之目的，藉之維持那布扶林(nalbuphine)的有效血中濃度大幅改善傳統習用注射劑型之使用量。 The present invention relates to a compound for treating acute or chronic pain in a mammal, a composition of a precursor soft drug, Sebacoyl dinalbuphine ester, and a pharmaceutical biodegradable polymer PLGA excipient. And the composition is prepared into a long-acting controlled release dosage form that can enter the human body. The use of macromolecules to achieve long-acting control of drug release, thereby maintaining the effective blood concentration of nalbuphine significantly improves the use of traditional conventional injectable dosage forms.

疼痛為一種感覺，由在身體末稍的痛覺受器(nociceptors)接受外來對動物體的機械、溫度及化學刺激，或身體內在的化學或電刺激，而引發痛覺神經衝動。這些衝動經由化學或神經痛覺路徑(nociceptive pathways)刺激大腦皮質或中樞神經，繼而造成疼痛的感覺。所有的脊椎動物甚至某些非脊椎動物如烏賊都有感知疼痛的能力，並從而產生對應的退縮及規避反應。疼痛若未加以控制或解除，則會對動物產生相當程度的緊迫(stress)，造成諸如ACTH，cortisol，ADH，catecholamine，glucose血中濃度的上升，以及insulin與testosterone的下降，進而影響心血管、呼吸、代謝，甚至組織癒合等生理功能的表現。疼痛反應是種保護性的反射系統，以警告個體正處於惡劣情況及組織受傷的狀態下。當病人有長期的疼痛時，長效止痛劑是有需求的，一長效作用鎮痛效果是特別為蒙受急性或慢性疼痛的病患所需要的。這些疼痛可能持續數天至數個月之久。舉例來說，急性疼痛，諸如手術後疼痛、外傷疼痛與燒傷疼痛，可能會持續4至6天；慢性疼痛，例如非惡性疼痛與癌症疼痛，可能持續數星期至數個月。類抗痛史，抗痛藥物的一大突破始於十九世紀末，科學家純化出阿斯匹靈與嗎啡，它們都是文獻記載中有效的鎮痛藥物。但是阿斯匹靈鎮痛效力不強，又有傷胃、抑制血小板凝集、胃出血之危險；而嗎啡止痛效果良好，可是副作用的餘慮造成使用上的諸多限制，因此兩者都需要於嚴密監控下謹慎使用。二十世 紀研發出的新藥，基本上仍然走阿斯匹靈、非類固醇抗發炎藥物(針對消炎止痛)與嗎啡(針對中樞神經系統中的嗎啡受體)的路線。 Pain is a sensation that involves the mechanical, thermal, and chemical stimuli of the animal's body, or the chemical or electrical stimuli inherent in the body, by the nociceptors at the end of the body, causing painful nerve impulses. These impulses stimulate the cerebral cortex or central nervous system via chemical or nociceptive pathways, which in turn causes a feeling of pain. All vertebrates and even certain invertebrates such as squid have the ability to perceive pain and thereby produce corresponding withdrawal and evasive responses. If the pain is not controlled or relieved, it will cause a considerable degree of stress on the animal, causing an increase in blood concentrations such as ACTH, cortisol, ADH, catecholamine, glucose, and a decrease in insulin and testosterone, which in turn affects the cardiovascular system. Performance of physiological functions such as breathing, metabolism, and even tissue healing. The pain response is a protective reflex system to warn individuals that they are in a bad condition and tissue injury. Long-acting analgesics are needed when the patient has long-term pain, and a long-acting analgesic effect is especially needed for patients suffering from acute or chronic pain. These pains can last from a few days to a few months. For example, acute pain, such as post-operative pain, traumatic pain, and burn pain, can last for 4 to 6 days; chronic pain, such as non-malignant pain and cancer pain, can last for weeks to months. A history of anti-pain, a major breakthrough in anti-pain drugs began in the late 19th century, when scientists purified aspirin and morphine, which are effective analgesics in the literature. However, aspirin is not effective in analgesic, but also has the risk of stomach injury, inhibition of platelet aggregation, and stomach bleeding. However, morphine has a good analgesic effect, but the side effects of the side effects cause many restrictions on the use, so both need to be closely monitored. Use with caution. Twenty world New drugs developed by Ji Ji are still basically taking the route of aspirin, non-steroidal anti-inflammatory drugs (for anti-inflammatory and analgesic) and morphine (for morphine receptors in the central nervous system).

目前市面上的類嗎啡止痛劑型效期短且副作用強，如蘇芬坦尼(Sufentanil)、瑞芬太尼(Remifentanil)、芬太尼(Fentanyl)、嗎啡(Morphin)注射劑。長期使用一天至少注射兩次到三次，產品使用不方便且藥物過量會產生呼吸抑制與藥物上癮等副作用，如何開發出長效且副作用輕微之類嗎啡止痛劑型是本發明探討的重點。那布扶林(Nalbuphine)為一種強力止痛劑可有效用於治療病患中度至重度疼痛。比較術後疼痛治療，那布扶林與嗎啡具同樣效果且副作用輕微，有較低的呼吸抑制及上癮等副作用，那布扶林藥理學分類屬於類嗎啡受體(Opioid receptor)kappa-活化劑且屬於Mu-receptor部分拮抗劑(Schmidt W.K. et al.,1985)，因此機轉會有界限效應(ceiling effect)如增加劑量大於30毫克不會造成進一步的呼吸抑制。因要跟上全世界製藥十大先進國家之腳步本國行政院院臺衛字第0960004264號公告那布扶林減列為非管制藥品。截至目前為止nalbuphine止痛效力之有效期間為三小時(血中濃度約10 ng/ml)到六小時(血中濃度約2.5 ng/ml)用此藥物治療(；病患必須住院且忍受多次注射，造成醫療資源浪費對病患更是痛苦。前驅軟藥(prosoft-drug)之設計能改善藥物的半衰期目前已廣泛應用於臨床且優良可行的。那布扶林為較親水性藥物(與前驅軟藥相比)與當利用長鏈碳酸進行酯化可增加了原始藥物脂溶性，可利用油性製劑或生物可降解材料包埋；而行肌肉注射時，在組織釋出率減緩而達到長效作用。而酯解酵素存在於各個組織器官中如血液、腦、肝、心、肺、腎、肌肉等，此前驅軟藥物之藥理作用及安全性則被報告和原藥完全相同。 Currently, morphine-like analgesics on the market have short duration and strong side effects, such as Sufentanil, Remifentanil, Fentanyl, and Morphin injections. Long-term use at least two to three times a day, the product is inconvenient to use and excessive drug production will produce side effects such as respiratory depression and drug addiction. How to develop long-acting and mild side effects such as morphine analgesic dosage form is the focus of the present invention. Nalbuphine is a powerful analgesic that is effective in treating moderate to severe pain in patients. Comparing postoperative pain treatment, Nabufolin has the same effect as morphine with mild side effects and low side effects such as respiratory depression and addiction. The pharmacological classification of Bufulin belongs to the class of morphine receptor kappa-activator. It belongs to the Mu-receptor partial antagonist ( Schmidt WK et al., 1985 ), so the machine has a ceiling effect such as increasing the dose greater than 30 mg without causing further respiratory depression. In order to keep up with the pace of the world's top ten pharmaceutical companies in the world, the National Executive Yuan, No. 0960004264, announced that it was reduced to non-controlled drugs. The effective period of nalbuphine analgesic efficacy has been treated with this drug for three hours (blood concentration of about 10 ng/ml) to six hours (blood concentration of about 2.5 ng/ml) (patients must be hospitalized and endured multiple injections) The waste of medical resources is even more painful for patients. The design of prosoft-drug can improve the half-life of drugs. It has been widely used in clinical practice and is excellent and feasible. Nabufolin is a more hydrophilic drug (with predecessors). Compared with the use of long-chain carbonic acid, the esterification of the original drug can increase the fat solubility of the original drug, and it can be embedded by oily preparation or biodegradable material. When the intramuscular injection is performed, the release rate of the tissue is slowed down to achieve long-term effect. The esterase is present in various tissues and organs such as blood, brain, liver, heart, lung, kidney, muscle, etc. The pharmacological action and safety of the previously driven soft drugs are reported to be identical to the original drug.

癸二醯雙那布扶林酯(SDE)是那布扶林(Nalbuphine)的一種前驅軟藥，能有效治療急性、慢性及手術後疼痛。本發明是以那布扶林前驅軟藥(Nalbuphine prosoft-drug)癸二醯雙那布扶林酯(SDE)及具生體相容性且可分解性之聚合物製備成長效控釋劑型。藥物製成長效控釋劑型之目的，是為 了使治療效果能比服用單劑量者維持更長的時間。其優點為可減少投藥次數，同時降低了病人忘記服藥的可能率，且由於投藥後可保持血中濃度之穩定而改善療效。需長期使用止痛藥劑之病人，希冀能夠脫離醫療機構有機會重回家庭或人群。本案發明人承襲本實驗室先前專利案-中華民國申請案號第089109293(新穎含那布扶林酯前驅藥(nalbuphine ester prodrug)口服醫藥組成物)之發明，鑑於傳統上無法大幅度將那布扶林(nalbuphin)效期延長，乃亟思加以改良創新且在中華民國申請案號第085106156(那布扶林多量酯)揭示前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)合成方法與化學結構，並經多年苦心孤詣潛心研究後，終於成功研發完成本件前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)、與藥學常用生物可分解高分子PLGA賦型劑之組成物，將該組合物製備成可進入體內之長效控釋劑型達到半個月或多月投藥一次之目標。 Shuangdibufolinide (SDE) is a precursor softener of Nalbuphine that is effective in treating acute, chronic and post-operative pain. The invention provides a growth-effective controlled release dosage form of Nalbuphine prosoft-drug, SDA and a biocompatible and decomposable polymer. The purpose of making a long-acting controlled release dosage form for a drug is The therapeutic effect can be maintained for a longer period of time than when a single dose is administered. The advantage is that the number of administrations can be reduced, and the possibility that the patient forgets to take the medicine is reduced, and the effect can be improved by maintaining the stability of the blood concentration after administration. Patients who need long-term use of analgesics can hope to get out of the medical institution and have the opportunity to return to their families or people. The inventor of the present invention inherited the previous patent case of the laboratory - the invention of the Republic of China application number 089109293 (the novel oral medicinal composition containing nalbuphine ester prodrug), in view of the traditional inability to substantially The effect of nalbuphin was prolonged, and it was improved and innovated by 亟思. In the Republic of China application No. 085106156 (Nabufulin polyester), the precursor softener, Sebacoyl dinalbuphine ester, was revealed. Synthetic method and chemical structure, and after many years of painstaking research, finally successfully developed this precursor soft drug - Sebacoyl dinalbuphine ester, and the commonly used biodegradable polymer PLGA The composition of the composition, which is prepared as a long-acting controlled release dosage form which can be administered into the body for a period of one or more months.

另外，在本案發明人雖已在知名國際期刊中揭示了前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)其它長效型控釋劑型-經皮給藥電動輔助方法(Jeng-Fen Huang et al.,2005)、注射用植物油針劑(Pao L.H.et al.,2005)，及其他那布扶林前驅軟藥如：單那布扶林癸酸酯(nalbuphine decanoate)、單那布扶林庚酸酯(nalbuphine enanthate)、單那布扶林戊酸酯(nalbuphine pivalate)、單那布扶林丙酸酯(nalbuphine propionate)，與本案使用藥學常用生物可分解高分子PLGA賦型劑之組成物製成植入劑(Sung K.C.et al.,1998)、控釋微球劑型(Fang-I.Liu et al.,2003)。但沒如預期達到半個月或多月投藥一次之目標，例如先前揭示，皮下注射內含那布扶林(50 mg/rabbit)單那布扶林丙酸酯(nalbuphine propionate)、單那布扶林戊酸酯(nalbuphine pivalate)、單那布扶林癸酸酯(nalbuphine decanoate)之PLGA控釋微球劑型(每組皆n=3)結果表明維持有效血中濃度2.5 ng/ml只能維持四天和目標有很大落差(Fang-I.Liu et al.,2003)推估是此類前驅軟藥的脂溶性不足，但後續發明人之研究【表一】，使用更高脂溶性前驅軟藥-癸二醯雙那 布扶林酯(Sebacoyl dinalbuphine ester)-PLGA長效控釋劑型，注射(150 mg/kg of SDE)在大鼠體內(n=7)，約注射那布扶林40 mg/rat。結果表明可運用PLGA高分子之特性(PLA/PGA組成比例、平均分子量等…)調控血中藥物濃度，可維持有效血中濃度2.5 ng/ml半個月或多月。因此基於實驗結果，本發明著重於用以治療哺乳動物急性或慢性疼痛之化合物、前驅軟藥癸二醯雙那扶林酯(Sebacoyl dinalbuphine ester)、與藥學常用生物可分解高分子PLGA賦型劑之組成物製成長效控釋劑型，可為新穎及進步性發現。 In addition, the inventors of this case have revealed in the well-known international journals that the long-acting controlled release dosage form of the precursor soft drug, Sebacoyl dinalbuphine ester, is a subcutaneous administration of electric assisted method ( Jeng -Fen Huang et al., 2005 ), injection vegetable oil injection ( Pao LHet al., 2005 ), and other naproxil precursor drugs such as: nalbuphine decanoate, monabu Nalbuphine enanthate, nalbuphine pivalate, nalbuphine propionate, and pharmaceutical biodegradable polymer PLGA excipient The composition was made into an implant ( Sung KC et al., 1998 ) and a controlled release microsphere formulation ( Fang-I. Liu et al., 2003 ). However, it was not expected to achieve the goal of one-half or more months of drug administration. For example, it was previously revealed that subcutaneous injection of nalbufolin (50 mg/rabbit) of nalbuphine propionate (sinnabu) The nalbuphine pivalate and nalbuphine decanoate PLGA controlled release microspheres (n=3 in each group) showed that the effective blood concentration of 2.5 ng/ml can only be maintained. Maintaining a four-day and target has a large gap ( Fang-I. Liu et al., 2003 ) estimates that this is a fat deficiency of the precursor soft drugs, but the subsequent inventors' research [Table 1], using higher fat solubility Precursor soft drug - Sebacoyl dinalbuphine ester - PLGA long-acting controlled release dosage form, injection (150 mg / kg of SDE) in rats (n = 7), about the injection of Nabufu Lin 40 mg/rat. The results show that the characteristics of the PLGA polymer (PLA/PGA composition ratio, average molecular weight, etc.) can be used to regulate the blood drug concentration, and the effective blood concentration can be maintained at 2.5 ng/ml for half a month or more. Therefore, based on the experimental results, the present invention focuses on a compound for treating acute or chronic pain in a mammal, a precursor soft drug, Sebacoyl dinalbuphine ester, and a commonly used biodegradable polymer PLGA excipient. The composition is formulated as a long-acting controlled release dosage form for novel and progressive discovery.

本發明之目的即在提供一種那布扶林長效控釋劑型，用以解決傳統上無法大幅度將那布扶林(nalbuphin)效期延長的問題，使治療效果能比服用單劑量者維持更長的時間。其優點為可減少投藥次數，同時降低了病人忘記服藥的可能率，且由於投藥後可保持血中濃度之穩定而改善療效。 The object of the present invention is to provide a long-acting controlled-release dosage form of nabufolium for solving the problem that the nalbuphin effect period cannot be extended greatly, so that the therapeutic effect can be maintained longer than that of taking a single dose. time. The advantage is that the number of administrations can be reduced, and the possibility that the patient forgets to take the medicine is reduced, and the effect can be improved by maintaining the stability of the blood concentration after administration.

為達成前述發明目的，本發明提供一種發展適當前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)長效控釋劑型。所使用藥學常用生物可分解高分子PLGA賦型劑之組成物，具有改善nalbuphine傳統習用注射劑型之使用量。本發明之另一目的係在於提供一種發展適當前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)長效控釋劑型、至少一藥學常用生物可分解PLGA賦型劑組合，其中該PLGA賦型劑包含至少以下之一：聚乳酸(PLA)、聚甘醇酸(PGA)、聚乳酸(PLA)或聚甘醇酸(PGA)的相關衍生物及其組合。其中聚乳酸(PLA)或聚甘醇酸(PGA)的相關衍生物可為聚琥珀酸丁二脂(PBS)、聚羥基脂肪酸酯(PHA)、聚己酸內酯(PCL)、聚羥基酯(PHB)、聚羥戊酯(PHV)、PHB和PHV的共聚物(PHBV)、聚乳酸(PLA)-聚乙二醇(PEG)共聚物(PLEG)。此PLGA賦型劑組合可運用高分子的特性調控藥物釋放快慢(PLA/PGA組成比例、平均分子量等)可使用於人體做為止痛使用，對於改善nalbuphine傳統習用注射劑型之使用量：試驗結果發現， 給予癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)SDE-PLGA(150 mg/kg of SDE)長效控釋劑型於SD大鼠，由不同的PLGA賦型劑組合調控血中藥物濃度且維持有效濃度2.5 ng/ml半個月或多月之目標，例如，使用PLGA(50：50 5k)(n=5)可維持有效濃度2.5 ng/ml約21天、而用PLGA(75：25 10k)(n=7)可維持有效濃度2.5 ng/ml約30天，脂溶性藥物於PLGA長效控釋劑基質內分子量大、PLA含量高藥物釋放的速度慢(PLA/PGA組成比例與分子量大小為：PLA 50^~100%/PGA 0^~50%，組成之分子量大小範圍：5 k^~20 K)。本發明經以體內實驗結果證實，當前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)合併使用藥學常用生物可分解高分子PLGA賦型劑之組成物可顯著改善nalbuphine傳統習用注射劑型之使用量。因此基於實驗結果可為新穎及進步性發現。本發明之目的即在於可達成上述發明目的之長效控釋劑型。本發明所提供之發展Sebacoyl dinalbuphine ester新劑型，亦可加入一藥學上可接受之賦形劑至該配方中，該配方可為稀釋劑、填充劑、結合劑、崩解劑、潤滑劑等至長效控釋劑型中，並且其長效控釋劑型可包含錠劑、膠囊、軟膠囊、丸劑、懸浮液、微球、口腔植入片、乳化針劑、植入劑及藥學上可行之長效控釋劑型。 In order to achieve the foregoing objects, the present invention provides a long-acting controlled release dosage form for developing a suitable precursor drug, Sebacoyl dinalbuphine ester. The composition of the commonly used biodegradable polymer PLGA excipient used in the pharmacy has the use of improving the conventional nalbuphine injection dosage form. Another object of the present invention is to provide a long-acting controlled release dosage form of a suitable precursor soft drug, Sebacoyl dinalbuphine ester, and at least one pharmaceutically-acceptable biodegradable PLGA excipient combination, wherein The PLGA excipient comprises at least one of the following: polylactic acid (PLA), polyglycolic acid (PGA), polylactic acid (PLA) or related derivatives of polyglycolic acid (PGA), and combinations thereof. The related derivatives of polylactic acid (PLA) or polyglycolic acid (PGA) may be polybutylene succinate (PBS), polyhydroxyalkanoate (PHA), polycaprolactone (PCL), polyhydroxyl Ester (PHB), polyhydroxyamyl ester (PHV), copolymer of PHB and PHV (PHBV), polylactic acid (PLA)-polyethylene glycol (PEG) copolymer (PLEG). The PLGA excipient combination can use the characteristics of the polymer to regulate the release rate of the drug (PLA/PGA composition ratio, average molecular weight, etc.), and can be used for the painless use of the human body, and to improve the use of the traditional nalbuphine injection dosage form: the test results are found , Sebacoyl dinalbuphine ester SDE-PLGA (150 mg/kg of SDE) long-acting controlled release dosage form was administered to SD rats, and the concentration of blood drug was adjusted by different PLGA excipient combinations. And maintain the effective concentration of 2.5 ng / ml for half a month or more, for example, using PLGA (50:50 5k) (n = 5) can maintain an effective concentration of 2.5 ng / ml for about 21 days, while using PLGA (75: 25 10k) (n=7) can maintain an effective concentration of 2.5 ng / ml for about 30 days, fat-soluble drugs in the PLGA long-acting controlled release matrix matrix, the molecular weight is large, PLA high drug release rate is slow (PLA / PGA composition ratio and The molecular weight is: PLA 50 ^~ 100% / PGA 0 ^~ 50%, the molecular weight of the composition range: 5 k ^~ 20 K). The invention has confirmed by in vivo experimental results that the current softening drug Sebacoyl dinalbuphine ester combined with the composition of the commonly used biodegradable polymer PLGA excipient can significantly improve the traditional use of nalbuphine. The amount of the injection dosage form used. Therefore, based on experimental results, it can be a novel and progressive discovery. The object of the present invention is to provide a long-acting controlled release dosage form which achieves the above object of the invention. The development of the new Sebacoyl dinalbuphine ester dosage form provided by the present invention may also be added to the formulation by a pharmaceutically acceptable excipient, which may be a diluent, a filler, a binder, a disintegrant, a lubricant, etc. Long-acting controlled release dosage form, and its long-acting controlled release dosage form may contain tablets, capsules, soft capsules, pills, suspensions, microspheres, oral implants, emulsified injections, implants, and pharmaceutically feasible long-acting effects. Controlled release dosage form.

本發明將就下列實施例作進一步說明，然該等實施例僅為例示說明之用，而不應被解釋為實施本發明之限制。 The invention is further illustrated by the following examples, which are intended to be illustrative only and not to be construed as limiting.

實施例一 SDE-PLGA長效控釋劑型配製Example 1 Preparation of SDE-PLGA long-acting controlled release dosage form

實驗設計及內容：Experimental design and content:

1. SDE-PLGA長效控釋劑型配製-以微球為例：(圖1)1. Preparation of SDE-PLGA long-acting controlled release dosage form - taking microspheres as an example: (Figure 1)

活性物質Active substance

癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester,SDE)是那布扶林(Nalbuphine)的一種前驅軟藥，前驅軟藥之設計能改善藥物的半衰期目前已廣泛應用於臨床且優良可行的。 Sebacoyl dinalbuphine ester (SDE) is a precursor softener of Nalbuphine. The design of the precursor softener can improve the half-life of the drug. It has been widely used in clinical practice and is excellent and feasible. .

賦形劑excipient

聚乳酸(polylactide,PLA)與聚甘醇酸(polyglycolide,PGA)依不同比例聚合而成，具良好之生物降解性及生物相容性。 Polylactide (PLA) and polyglycolide (PGA) are polymerized in different proportions, and have good biodegradability and biocompatibility.

i.取一個乾淨樣品瓶加入PLGA高分子900 mg、活性成分SDE粉末1,200 mg，加入有機溶劑二氯甲烷9 ml(放入攪拌磁石協助溶解)此為油相。 i. Take a clean sample vial and add 900 mg of PLGA polymer, 1,200 mg of active ingredient SDE powder, and add 9 ml of organic solvent dichloromethane (dissolved in a stirring magnet to assist dissolution). This is the oil phase.

ii.水相為0.5% PVA 900 ml。先將水相注入恆溫夾套反應槽中(維持環境溫度為8-10℃，溫度過高藥物會降解且易有孔洞的產生)，以均質機攪拌轉速為1,800 RPM。 Ii. The aqueous phase is 0.5% PVA 900 ml. The water phase is first injected into the thermostated jacket reaction tank (maintaining an ambient temperature of 8-10 ° C, the temperature is too high, the drug will degrade and easy to have pores), and the homogenizer stirring speed is 1,800 RPM.

iii.使用氣密針吸取油相以等速注入水相中且利用剪切應力使微球成型(均質時間為4 min)之後於20℃攪拌(轉速300 RPM)2小時使微球固化後在30℃攪拌(轉速300 RPM)2小時使有機溶劑揮發去除。 Iii. Aspirate the oil phase with a gas-tight needle and inject it into the water phase at a constant rate and shape the microspheres by shear stress (homogenization time is 4 min), then stir at 20 ° C (300 RPM) for 2 hours to cure the microspheres. The organic solvent was volatilized and removed by stirring at 30 ° C (300 RPM) for 2 hours.

iv.過篩(上篩150 μm；下篩35 μm)去除水相並凍乾(凍乾條件：-40.0℃ 1.0x10^-1 psi)可得SDE-PLGA微球。 Iv. Sifting (150 μm on the sieve; 35 μm on the lower sieve) The aqueous phase was removed and lyophilized (freeze-dried conditions: -40.0 ° C 1.0 x 10 ^-1 psi) to obtain SDE-PLGA microspheres.

調配物實例1 Formulation example 1

調配物實例2(PLGA分子量5kDa) Formulation Example 2 (PLGA molecular weight 5 kDa)

調配物實例3(PLGA分子量10kDa) Formulation Example 3 (PLGA molecular weight 10 kDa)

調配物實例4(PLGA分子量15kDa) Formulation Example 4 (PLGA molecular weight 15 kDa)

調配物實例5(PLGA分子量18kDa) Formulation Example 5 (PLGA molecular weight 18 kDa)

調配物實例6(PLGA分子量20kDa) Formulation Example 6 (PLGA molecular weight 20 kDa)

調配物實例7(PLGA分子量10kDa) Formulation Example 7 (PLGA molecular weight 10 kDa)

2. SDE-PLGA藥物包埋率(Drug loading %)與包埋成功率(Encapsulation %)測定-以微球為例：2. SDE-PLGA drug embedding rate (Drug loading %) and embedding success rate (Encapsulation %) determination - taking microspheres as an example:

i.秤取SDE-PLGA微球(>10 mg)加入乙氰溶解為毫克微球/毫升乙氰之溶液(例如：秤取SDE-PLGA微球11.2 mg，需加入11.2 ml乙氰)N=3。 i. Weigh SDE-PLGA microspheres (>10 mg) and add acetonitrile to dissolve the solution into milligrams of microspheres/ml of acetonitrile (for example: weigh 11.2 mg of SDE-PLGA microspheres, add 11.2 ml of acetonitrile) N= 3.

ii.將上述溶解液以乙氰稀釋100倍後取100 μl注射入HPLC中，其結果用檢量線定量。 Ii. The above solution was diluted 100 times with acetonitrile, and 100 μl was injected into HPLC, and the results were quantified by a calibration curve.

iii.藥物包埋率(Drug loading%)=(API/(API+PLGA))^＊100% Iii. Drug embedding rate (Drug loading%) = (API / (API + PLGA)) ^* 100%

iv.包埋成功率(Encapsulation %)=(藥物包埋率/理論藥物包埋率)^＊100% Iv. Embedding success rate (Encapsulation %) = (drug embedding rate / theoretical drug embedding rate) ^* 100%

3.雷射粒徑分析：3. Laser particle size analysis:

i.取適量的微球加入水中再緩慢滴入樣品槽內，用樣品槽內攪拌器加以攪拌使微球均勻分散於水中， i. Take an appropriate amount of microspheres into water and slowly drip into the sample tank, and stir them with a stirrer in the sample tank to uniformly disperse the microspheres in water.

ii.將此樣品槽以He-Ne鐳射光源(633 nm)之紅外光散射分析儀中，以測定微球的粒徑分佈。 Ii. The sample cell was analyzed by an infrared light scattering analyzer of a He-Ne laser source (633 nm) to determine the particle size distribution of the microspheres.

iii.重複實驗三次將所得數據求其平均值與標準差 Iii. Repeat the experiment three times to obtain the average and standard deviation of the obtained data.

實驗結果：Experimental results:

使用(50：50 5 k、75：25 10 k、75：25 15 k、75：25 18 k、75：25 20 k、100：0 10 k)上述六種生物可分解高分子PLGA賦型劑在相同均質轉速1800 RPM與相同API與高分子比例400 mg/300 mg，研究藥物的包埋率，包埋成功率與平均粒徑改變量為何，結果表明在相同轉速與相同API與高分子比例(400 mg/300 mg)，在不同高分子材料情況下藥物包埋率(45-60%)與平均粒徑大小(60-70 μm)R及包埋成功率(81-100%)沒有顯著的改變【表二】。 Use (50:50 5 k, 75:25 10 k, 75:25 15 k, 75:25 18 k, 75:25 20 k, 100:0 10 k) the above six biodegradable polymer PLGA excipients At the same homogenization speed of 1800 RPM and the same API to polymer ratio of 400 mg / 300 mg, the embedding rate of the study drug, the embedding success rate and the average particle size change, the results show that the same speed and the same API and polymer ratio (400 mg/300 mg), the drug embedding rate (45-60%) and average particle size (60-70 μm) R and embedding success rate (81-100%) were not significant in the case of different polymer materials. Change [Table 2].

實施例二 SDE-PLGA長效控釋劑型體外溶離試驗Example 2 In vitro dissolution test of SDE-PLGA long-acting controlled release dosage form

1.體外溶離試驗-以微球為例：1. In vitro dissolution test - taking microspheres as an example:

i.將16X125mm螺旋帽試管清洗乾淨並滅菌，且將體外溶離緩衝液0.025 M PBS+0.5%Tween20 400 ml配置於500 ml血清瓶內並滅菌且內含攪拌磁石。 i. The 16X125mm screw cap test tube was cleaned and sterilized, and the in vitro dissolution buffer 0.025 M PBS + 0.5% Tween 20 400 ml was placed in a 500 ml serum bottle and sterilized and contained a stirring magnet.

ii.在無菌操作臺將微球粉末(不能滅菌會使微球性質改變-約5 mg)倒入400 ml溶離緩衝液中，放置在磁石攪拌器800 RPM高速攪拌使微球均勻懸浮於溶離緩衝液中。 Ii. Pour the microsphere powder (not sterilized to change the properties of the microspheres - about 5 mg) into 400 ml of dissolving buffer at the aseptic table, place it in a magnet stirrer at 800 RPM and stir at high speed to suspend the microspheres in the dissolution buffer. In the liquid.

iii.吸取5 ml懸浮液注入16X125mm螺旋帽試管中，分別標上時間(N=3)。將分裝好的試管螺旋帽封口貼上透明膠帶(防止水浴槽內水濺入污染且防止長時間搖晃螺旋帽脫落)。 Iii. Pipette 5 ml of the suspension into a 16X125mm screw cap test tube and mark the time (N=3). Attach the sealed test tube screw cap to the transparent tape (prevent the water from splashing into the water bath and prevent the screw cap from falling off for a long time).

iv.之後放置於水浴槽中37℃，100 RPM反應時間到取出。 Iv. After placing in a water bath at 37 ° C, 100 RPM reaction time to take out.

v.本方法主要研究微球內部藥物釋出量，將標示不同天試管吸取上清液4.5 ml，加入乙氰4.5 ml破壞微球，震搖10 min。微球完全溶解後，樣品過0.45μm薄膜(PVDF)後吸取200 μl上清液直接注入HPLC系統當中且結果利用公式藥物釋出量(%)=100(%)-藥物剩餘量(%)。 v. This method mainly studies the amount of drug released inside the microsphere. It will mark 4.5 ml of the supernatant from the test tube in different days, and add 4.5 ml of acetonitrile to destroy the microspheres and shake for 10 min. After the microspheres were completely dissolved, the sample was passed through a 0.45 μm film (PVDF), and 200 μl of the supernatant was aspirated directly into the HPLC system and the results were released using the formula drug (%) = 100 (%) - drug remaining amount (%).

2.高效能液相層析儀分析條件：2. High performance liquid chromatography analysis conditions:

用於體外實驗之分析如SDE-PLGA藥物包埋率(Drug loading%)與包埋成功率(Encapsulation %)測定、與體外溶離試驗，準確定量藥物濃度。 For in vitro experiments such as SDE-PLGA drug implantation rate (Drug loading%) and embedding success rate (Encapsulation %) determination, and in vitro dissolution test, accurate quantitative drug concentration.

移動相(mobile phase)是由25%醋酸鈉緩衝液(5毫莫耳/公升)及75%乙氰(ACN)加上0.07%(體積百分比V/V)與0.1%三乙胺(Triethylamine)組成，層析柱(column)為Thermo Hypersil Betasil Silica 150x4.6毫米，3微米，溫度為攝氏30度，紫外光偵測器所使用的波長為210 nm，使用的流速為每分鐘1.3毫升，分析樣品時間為每7分鐘一個樣品。 The mobile phase is composed of 25% sodium acetate buffer (5 mmol/L) and 75% acetonitrile (ACN) plus 0.07% (v/v) and 0.1% triethylamine. Composition, column is Thermo Hypersil Betasil Silica 150x4.6 mm, 3 microns, temperature is 30 degrees Celsius, UV detector uses a wavelength of 210 nm, using a flow rate of 1.3 ml per minute, analysis The sample time is one sample every 7 minutes.

3.檢量線之配製：3. Preparation of the calibration line:

將前驅軟藥-癸二醯雙那布扶林酯與原始藥物-那布扶林混合配製成250、500、1000、1500、2000、3500、5000、7500、10000(ng/ml)的濃度以標準液配製在乙氰(ACN)中。 The precursor soft drug, bismuth shuangfufufufen, was mixed with the original drug, nabufuran, to a concentration of 250, 500, 1000, 1500, 2000, 3500, 5000, 7500, 10000 (ng/ml). Formulated in acetonitrile (ACN) as a standard solution.

經高效能液相層析儀分析後，以層析圖譜中癸二醯雙那布扶林酯與那布扶林之各別波峰面積值與其相對濃度作圖，可得到兩校正曲線，並以標準偏差(Standard deviation，SD)、變異係數(% CV)及誤差(% error)來驗證精確性與準確性。 After analysis by high performance liquid chromatography, the respective peak area values of the bismuth shuangbufulin and nabufulin in the chromatogram are plotted, and two calibration curves can be obtained. Standard deviation (SD), coefficient of variation (% CV), and error (% error) to verify accuracy and accuracy.

實驗結果：Experimental results:

比較相同PLA/PGA比例、不同分子量(75/25 10k)、(75/25 15k)、(75/25 18k)、(75/25 20k)SDE-PLGA微球此時比較溶離結果，發現分子量越低藥物釋放越快。三十天藥物釋放為75/25 10k(87.71%±13.81%)、75/25 15k(56.64%±6.40%)、75/25 18k(57.30%±14.33%)、75/25 20k(42.78%±5.42%)數據為(mean±SD，n=3)【圖2】。比較不同PLA/PGA比例、相同分子量(75/25 10k)、(100/0 10k)SDE-PLGA微球此時比較溶離結果、發現PLA含量越低時低藥物釋放越快三十天藥物釋放為75/25 10k(87.71%±13.81%)、100/0 10k(28.16%±6.31%)數據為(mean±SD，n=3)【圖3】。比較不同PLA/PGA比例、不同分子量(50/50 6k)、(75/25 10k)SDE-PLGA微球此時比較溶離結果、發現結果與上述相同，發現分子量越低且PLA含量越低時藥物釋放越快三十天藥物釋放為75/25 10k 100/0 10k(88.54%±6.47%)、(87.71%±13.81%)數據為(mean±SD，n=3)【圖4】。 Comparing the same PLA/PGA ratio, different molecular weight (75/25 10k), (75/25 15k), (75/25 18k), (75/25 20k) SDE-PLGA microspheres at this time, the dissolution results were compared, and the molecular weight was found to be The faster the drug is released. Drug release for 30 days was 75/25 10k (87.71% ± 13.81%), 75/25 15k (56.64% ± 6.40%), 75/25 18k (57.30% ± 14.33%), 75/25 20k (42.78% ± 5.42%) The data is (mean±SD, n=3) [Fig. 2]. Compare different PLA/PGA ratios, the same molecular weight (75/25 10k), (100/0 10k) SDE-PLGA microspheres at this time to compare the dissolution results, and find that the lower the PLA content, the faster the drug release is 30 days. 75/25 10k (87.71%±13.81%), 100/0 10k (28.16%±6.31%) data is (mean±SD, n=3) [Fig. 3]. Comparing different PLA/PGA ratios, different molecular weights (50/50 6k), (75/25 10k) SDE-PLGA microspheres at this time, the results of the dissolution were the same as above, and the lower the molecular weight and the lower the PLA content, the drugs were found. The faster the release, the 30-day drug release was 75/25 10k 100/0 10k (88.54% ± 6.47%), and (87.71% ± 13.81%) data was (mean ± SD, n = 3) [Fig. 4].

實施例三 SDE-PLGA長效控釋劑型體內藥物動力學試驗Example 3 In vivo pharmacokinetic test of SDE-PLGA long-acting controlled release dosage form

實驗設計及內容：Experimental design and content:

研究的主要目的在測定及比較大白鼠在給予單一劑量之不同處方、劑型，藥物在大白鼠體內之吸收、分佈及排除情形。經由分析單一劑量 SDE-PLGA長效控釋劑肌肉注射後大白鼠之血漿中之Nalbuphine及SDE，即可得知不同劑型在大白鼠體內之吸收、代謝及排除情形，並可證明所篩選配方達到一個月有效之目的。 The main purpose of the study was to determine and compare the absorption, distribution and elimination of different prescriptions, dosage forms and drugs in rats in a single dose. By analyzing a single dose Nalbuphine and SDE in the plasma of rats with SDE-PLGA long-acting controlled release agent after intramuscular injection can be used to know the absorption, metabolism and elimination of different dosage forms in rats, and it can be proved that the selected formula is effective for one month. purpose.

1.小型動物大白鼠肌肉注射SDE-PLGA長效控釋劑型之藥動學研究1. Pharmacokinetic study of intramuscular injection of SDE-PLGA long-acting controlled release dosage form in small animal rats

實驗動物：Experimental animals:

i.種類：大白鼠(Rat)。 i. Type: Rat (Rat).

ii.品系：Sprage-Dawley。 Ii. Line: Sprage-Dawley.

iii.來源：國科會實驗動物中心。 Iii. Source: National Laboratory Animal Center.

iv.開始實驗鼠齡：年輕成鼠，6-9週大。 Iv. Start experimentation Rat age: young adult rats, 6-9 weeks old.

v.開始實驗體重：大白鼠之體重範圍越小越好，最好之體重範圍為200-300克。 v. Start experiment weight: The smaller the weight range of the rats, the better. The best weight range is 200-300 grams.

vi.標示方式：在尾部標記。 Vi. Marking method: mark at the tail.

vii.實驗數量：每組6-7隻雌鼠。 Vii. Number of experiments: 6-7 female mice per group.

實驗期間：大白鼠給藥後將被觀察並收集各種試樣直至nalbuphine偵測不到為止(約一個小月)。 During the experiment: The rats will be observed and collected after administration until the nalbuphine is not detected (about one small month).

給藥途徑與方法：主要以肌肉(右大腿)注射SD-PLGA長效控釋劑將根據體重調整劑量，以單一劑量方式給藥。 Routes and methods of administration: The SD-PLGA long-acting controlled release agent is mainly injected into the muscle (right thigh) to adjust the dose according to the body weight, and is administered in a single dose.

藥物劑型之分析：製備好的劑型必須分析其化學強度，也就是製劑必須要有相當好的均一性及穩定性。 Analysis of pharmaceutical dosage forms: The prepared dosage form must be analyzed for its chemical strength, that is, the formulation must have a fairly good homogeneity and stability.

方法設計：Method design:

i.動物之適應與選擇：大約25隻雌鼠在抵達實驗室後，必須經過至少一週之適應期與觀察期，在此期間動物將被觀察其一般的健康狀況及有無任何疾病徵狀。動物均由供應者給予適當的疫苗。在實驗開始前動物均由獸醫師做一完全的身體檢查。 i. Adaptation and selection of animals: Approximately 25 females must undergo at least one week of acclimatization and observation after arrival in the laboratory, during which time the animals will be observed for their general health and with or without any signs of disease. Animals are given appropriate vaccines by the supplier. Animals were thoroughly examined by a veterinarian before the start of the experiment.

ii.動物之選擇：在經過觀察期後，若有任何疾病徵狀或有任何重要之不正常生理現象的大白鼠均將淘汱。 Ii. Selection of animals: After the observation period, any rats with any disease symptoms or any important abnormal physiological phenomena will be scoured.

iii.動物飼養環境：大白鼠將被飼養在溫度控制在攝氏20-30度、濕度控制在百分之30至70的環境中，另外，並提供每天12小時的照光。 Iii. Animal feeding environment: The rats will be kept in an environment where the temperature is controlled at 20-30 degrees Celsius and the humidity is controlled at 30 to 70 percent. In addition, 12 hours of illumination per day is provided.

iv.動物飼料：飼料為Purina Mills公司出品品質保証之Rodent Chow^® #5002，飼料為無限制供應，且在實驗前不需禁食。 Iv. Animal feed: The feed is Rodent Chow ^® #5002, which is guaranteed by Purina Mills. The feed is unrestricted and does not require fasting before the experiment.

v.飲水：飲水為無限制供應。 v. Drinking water: Drinking water is an unlimited supply.

vi.隨機試驗方式：動物之分組以體重為分類標準採用Xybion隨機程式隨機分組，使各實驗組的大白鼠體重分佈均勻。動物分為兩組，每組至少有6隻雌鼠。 Vi. Randomized trials: Animals were grouped by weight using the Xybion randomization program, and the rats in each experimental group were evenly distributed. Animals were divided into two groups with at least 6 female rats in each group.

vii.給藥：給含藥為150 mg/kg SDE SDE-PLGA微球(懸浮於1.25%CMC中)。給藥方式是右大腿肌肉注射。 Vii. Administration: 150 mg/kg SDE SDE-PLGA microspheres (suspended in 1.25% CMC) were administered. The mode of administration is intramuscular injection of the right thigh.

viii.試樣收集：血漿 Viii. Sample collection: plasma

本研究將以尾靜脈取血方式自大白鼠取得血樣，將大白鼠單獨放置在一限制裝置中以減低給藥及取樣時對大白鼠所造成的壓力，之後，給予150 mg/kg SDE-PLGA微球。給藥後0.5,1,2,6,24,30,48,54,72,96,102,120,168,240小時與12,14,16,18,20,24,26,28天自大白鼠尾靜脈取血0.5 ml並加入適量之肝素(1/10體積)。取全血後離心取上清液血漿，所有血漿樣均儲存在-80℃之冰箱中。在實驗中任何時間，若發現有任何動物病危，則基於人道理由，將以二氧化碳使其安樂死。 In this study, blood samples were taken from rats by tail vein blood sampling, and the rats were placed in a restriction device alone to reduce the pressure on the rats during administration and sampling. After that, 150 mg/kg SDE-PLGA was administered. Microspheres. 0.5,1,2,6,24,30,48,54,72,96,102,120,168,240 hours and 12,14,16,18,20,24,26,28 days after administration, 0.5 ml of blood was taken from the tail vein of rats. Add an appropriate amount of heparin (1/10 volume). After taking whole blood, the supernatant plasma was centrifuged, and all plasma samples were stored in a -80 ° C refrigerator. At any time during the experiment, if any animal is found to be in critical condition, it will be euthanized with carbon dioxide for humanitarian reasons.

試樣分析：各種試樣將以已開發之超效能液相層析法質譜分析(UPLC/Ms/Ms)其中之Nalbuphine及SDE。 Sample analysis: Various samples will be analyzed by mass spectrometry mass spectrometry (UPLC/Ms/Ms) developed by Nalbuphine and SDE.

資料分析：資料圖形及表格將包含所給藥物在血漿內之變化情形，另外，將以軟體計算Nalbuphine及SDE在大白鼠體內之各種藥動學參數，包括相對生體可用率。 Data analysis: The data graph and table will contain changes in the plasma of the given drug. In addition, various pharmacokinetic parameters of Nalbuphine and SDE in the rat, including relative bioavailability, will be calculated in software.

2. UPLC/Ms/Ms分析條件：2. UPLC/Ms/Ms analysis conditions:

採用超高效能液相層析串聯質譜儀Ultra Performance Liquid Chromatography(UPLC)：/M ass/Mass UPLC/MS/MS,(API 3000 Applied biosystens，美國)三節四極質(triple-quadrupole)譜儀配備有離子噴射(ionspray)LC/MS interface)進行分析：層析柱(Column)為ACQUITY UPLC/BEH HILIC/2.1 X 100毫米/1.73微米/40度C；移動相(Mobile phase)為2 mM乙酸銨(Ammonium acetate)及0.1%甲酸(formic acid)於水(13%)和乙晴(Acetonitrile)(87%)；流速(Flow)為每分鐘0.25毫升；注射體積(Injection volume)為5微升。分析癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)選用的母離子是441.5(m/z)、子離子是423.5(m/z)；分析那不扶林(Nalbuphine)選用的母離子均為358.3(m/z)、子離子均為340.3(m/z)。分析內標物：乙基嗎啡(Ethylmorphine)選用的母離子均為314.2(m/z)、子離子均為183.1(m/z)；烯丙羥嗎啡酮(Naloxone)選用的母離子均為328.3(m/z)、子離子均為310.3(m/z)。 Ultra-performance liquid chromatography tandem mass spectrometer Ultra Performance Liquid Chromatography (UPLC): /M ass/Mass UPLC/MS/MS, (API 3000 Applied biosystens, USA) three-triple-quadrupole spectrometer equipped with Ion-jet (ionspray) LC/MS interface) analysis: Column (AC) is ACQUITY UPLC/BEH HILIC/2.1 X 100 mm / 1.73 μm / 40 ° C; Mobile phase (Mobile phase) is 2 mM ammonium acetate ( Ammonium acetate) and 0.1% formic acid in water (13%) and Acetonitrile (87%); flow rate (Flow) was 0.25 ml per minute; injection volume was 5 microliters. The parent ion selected from Sebacoyl dinalbuphine ester was 441.5 (m/z) and the daughter ion was 423.5 (m/z). The parent ions selected for Nalbuphine were analyzed. It was 358.3 (m/z) and the product ions were 340.3 (m/z). The internal standard of the analysis: Ethylmorphine used 314.2 (m / z), the daughter ion was 183.1 (m / z); the parent ion selected by naloxone (Naloxone) was 328.3 (m/z) and the daughter ions were both 310.3 (m/z).

3.血漿樣品處理：3. Plasma sample processing:

i.於16x125mm試管底部加入內部標準品(Ethyl-morphine 2 μg/ml & Naloxone 200 ng/ml 50 μl)。 i. Add an internal standard (Ethyl-morphine 2 μg/ml & Naloxone 200 ng/ml 50 μl) to the bottom of a 16 x 125 mm tube.

ii.加入0.5 N Na₂CO₃ pH=10.050 μl(鹼性藥物由離子態還原成分子態)。 Ii. Add 0.5 N Na ₂ CO ₃ pH=10.050 μl (the basic drug is reduced by the ionic state).

iii.加入萃取液(Ether：DCM=7：3)4 ml且放置冷房4℃並放置冰浴上(防止前驅軟藥SDE於血漿中降解)。 Iii. Add 4 ml of extract (Ether: DCM = 7:3) and place in a cold room at 4 ° C and place on an ice bath (to prevent degradation of the precursor soft drug SDE in plasma).

iv.加入血漿100 μl，且震盪10 min之後離心3,000 RPM 10 min。 Iv. Add 100 μl of plasma and centrifuge for 3,000 RPM for 10 min after shaking for 10 min.

v.於-80℃冰箱將下水層凝固，並將上清液倒入新試管內13x100mm。 v. The lower aqueous layer was solidified in a refrigerator at -80 ° C, and the supernatant was poured into a new test tube 13 x 100 mm.

vi.氮氣吹乾，壓力10 psi、40℃、20 min(時間過長前驅軟藥SDE可能受熱降解)。 Vi. Blow dry with nitrogen at a pressure of 10 psi, 40 ° C, 20 min (the long-term pro-soft drug SDE may be thermally degraded).

vii.將200μl稀釋液(ACN：Q水=85：15)加入試管內回溶並注射5μl至UPLC/Ms/Ms中。 Vii. 200 μl of the dilution (ACN: Q water = 85:15) was added to the test tube for reconstitution and 5 μl was injected into UPLC/Ms/Ms.

4.檢量線之配製：4. Preparation of the calibration line:

將前驅軟藥-癸二醯雙那布扶林酯與原始藥物-那布扶林混合配製成10、25、50、100、250、500、1000、2500、5000(ng/ml)的濃度以標準液配製在乙氰(ACN)中且取10 μl加入90 μl空白血漿(稀釋10倍)中後樣品處理。 The precursor soft drug, quinone diterpene furfural, was mixed with the original drug, nabufulin, to a concentration of 10, 25, 50, 100, 250, 500, 1000, 2500, 5000 (ng/ml). The sample was treated with standard solution in acetonitrile (ACN) and 10 μl was added to 90 μl of blank plasma (diluted 10 times).

經UPLC/Ms/Ms分析後，以層析圖譜中癸二醯雙那布扶林酯與那布扶林之各別波峰面積值與其相對濃度作圖，可得到兩校正曲線，並以標準偏差(Standard deviation，SD)、變異係數(% CV)及誤差(% error)來驗證精確性與準確性。 After UPLC/Ms/Ms analysis, the respective peak area values of the bismuth shuangbufulin and nabufulin in the chromatogram were plotted, and two calibration curves were obtained, and the standard deviation was obtained. (Standard deviation, SD), coefficient of variation (% CV), and error (% error) to verify accuracy and accuracy.

實驗結果：Experimental results:

結果表明可由調控高分子性質(PLA/PGA比例、平均分子量)來調控血中藥物濃度且結果與理論吻合，藥物釋放速度(50：50 5k)>(75：25 10k)>(75：25 18k)，可調控血中藥物濃度且維持有效濃度2.5 ng/ml半個月或多月之目標【圖5】；藥物釋放量分別為(50：50 5k)>(75：25 10k)>(75：25 18k)，可知由PLA/PGA比例、平均分子量來調控血中藥物濃度且結果與理論吻合，各項參數如表三。 The results showed that the concentration of the drug in the blood could be regulated by the properties of the regulated polymer (PLA/PGA ratio, average molecular weight) and the results were in agreement with the theory. The drug release rate (50:50 5k)>(75:25 10k)>(75:25 18k) ), can regulate the concentration of blood drug and maintain the effective concentration of 2.5 ng / ml for half a month or more [Figure 5]; drug release is (50:50 5k)> (75:25 10k)> (75 :25 18k), it can be seen that the concentration of PLA/PGA and the average molecular weight regulate the concentration of blood drugs and the results are in agreement with the theory. The parameters are shown in Table 3.

綜上所述，本案前驅軟藥-癸二醯雙那布扶林酯(Sebacoyl dinalbuphine ester)、與藥學常用生物可分解高分子PLGA賦型劑之組成物，並將該組合物製備成可進入人體之長效控釋劑型。藉由高分子的而達到長效控制藥物釋放之目的，藉之維持那布扶林(nalbuphine)的有效血中濃度大幅改善傳統習用注射劑型之使用量。應已充分符合專利新穎性及進步性之法定發明專利要件，爰依法提出申請，懇請 貴局核准本件發明專利申請案，以勵發明，至感德便。 In summary, the composition of the precursor soft drug, Sebacoyl dinalbuphine ester, and the commonly used biodegradable polymer PLGA excipient, and the composition is prepared to enter Long-acting controlled release dosage form of the human body. The use of macromolecules to achieve long-acting control of drug release, thereby maintaining the effective blood concentration of nalbuphine significantly improves the use of traditional conventional injectable dosage forms. It should have fully complied with the statutory invention patent requirements for patent novelty and progress, and file an application according to law. You are requested to approve the application for this invention patent to encourage invention.

圖1、範例：微球-長效控釋劑型 Figure 1. Example: Microspheres - Long-acting controlled release dosage form

圖2、實驗組(75/25 10k)、(75/25 15k)、(75/25 18k)、(75/25 20k)長效控釋劑型之溶離情況，比較不同分子量藥物釋出情形(mean±SD，n=3)。 Figure 2. Dissolution of long-acting controlled release dosage forms of experimental groups (75/25 10k), (75/25 15k), (75/25 18k), (75/25 20k), comparing the release of different molecular weight drugs (mean ±SD, n=3).

圖3、實驗組(75/25 10k)、(100/0 10k)長效控釋劑型之溶離情況(mean±SD，n=3)，比較不同PLA含量藥物釋出情形。 Figure 3. The dissolution of the experimental group (75/25 10k), (100/0 10k) long-acting controlled release dosage form (mean±SD, n=3), comparing the drug release of different PLA content.

圖4、實驗組(50/50 5k)、(75/25 10k)長效控釋劑型之溶離情況(mean±SD，n=3)比較不同分子量且不同PLA含量藥物釋出情形。 Figure 4. The dissolution of the experimental group (50/50 5k), (75/25 10k) long-acting controlled release dosage form (mean±SD, n=3) compared the drug release of different molecular weights and different PLA content.

圖5、體內中那布扶林(Nalbuphine)藥物濃度與時間做圖，給予實驗動物SD-rat肌肉注射長效控釋劑型50：50 5k(150 mg/kg SDE)、75：25 10k(150 mg/kg SDE)、75：25 18k(150 mg/kg SDE)。(A)笛卡兒座標圖(B)半對數座標圖。 Figure 5. Nabuffin (Nalbuphine) drug concentration and time in the body, given experimental animals SD-rat intramuscular injection of long-acting controlled release dosage form 50:50 5k (150 mg / kg SDE), 75:25 10k (150 Mg/kg SDE), 75:25 18k (150 mg/kg SDE). (A) Cartesian coordinate map (B) semi-logarithmic coordinate map.

Claims (6)

一種用以改善那布扶林(nalbuphine)注射劑型之使用量之一長效控釋劑型醫藥組合物，該組合物包含：(a)一前驅軟藥-癸二醯雙那布扶林酯化合物；及(b)至少一藥學常用生物可分解高分子PLGA賦型劑之組成物。 A long-acting controlled-release dosage form pharmaceutical composition for improving the use amount of nalbuphine injection preparation, the composition comprising: (a) a precursor soft drug - bismuth shuangbufulin ester compound And (b) a composition of at least one pharmaceutically acceptable biodegradable polymeric PLGA excipient. 如申請專利範圍第1項所述之醫藥組合物，進一步包含該化合物在藥學上可接受之鹽類、溶劑合物或在醫藥上具有藥學功能之相關衍生物。 The pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable salt, solvate or a pharmaceutically acceptable derivative of the compound. 如申請專利範圍第1項所述之醫藥組合物，其中該PLGA賦型劑包含至少以下之一：聚乳酸(PLA)、聚甘醇酸(PGA)、聚乳酸(PLA)或聚甘醇酸(PGA)的衍生物及其組合。 The pharmaceutical composition according to claim 1, wherein the PLGA excipient comprises at least one of polylactic acid (PLA), polyglycolic acid (PGA), polylactic acid (PLA) or polyglycolic acid. (PGA) derivatives and combinations thereof. 如申請專利範圍第3項所述之醫藥組合物，其中該PLA/PGA組成比例與分子量大小為：PLA 50^~100%/PGA 0^~50%，組成之分子量大小範圍：5 k^~20 K。 The pharmaceutical composition according to claim 3, wherein the PLA/PGA composition ratio and molecular weight are: PLA 50 ^~ 100% / PGA 0 ^~ 50%, and the molecular weight range of the composition ranges from 5 k ^to 20 K. 如申請專利範圍第3項所述之醫藥組合物，其中該聚乳酸(PLA)或聚甘醇酸(PGA)的衍生物可為聚琥珀酸丁二脂(PBS)、聚羥基脂肪酸酯(PHA)、聚己酸內酯(PCL)、聚羥基酯(PHB)、聚羥戊酯(PHV)、PHB和PHV的共聚物(PHBV)、聚乳酸(PLA)-聚乙二醇(PEG)共聚物(PLEG)。 The pharmaceutical composition according to claim 3, wherein the polylactic acid (PLA) or polyglycolic acid (PGA) derivative is polybutylene succinate (PBS) or polyhydroxyalkanoate ( PHA), polycaprolactone (PCL), polyhydroxyester (PHB), polyhydroxyamyl ester (PHV), copolymer of PHB and PHV (PHBV), polylactic acid (PLA)-polyethylene glycol (PEG) Copolymer (PLEG). 如申請專利範圍第1項所述之醫藥組合物，進一步包含一長效控釋劑型，其中該劑型為以下之一：錠劑、膠囊、軟膠囊、丸劑、懸浮液、微球、口腔植入片、乳化針劑、植入劑及藥學上可行之長效控釋劑型。 The pharmaceutical composition according to claim 1, further comprising a long-acting controlled release dosage form, wherein the dosage form is one of the following: a tablet, a capsule, a soft capsule, a pill, a suspension, a microsphere, an oral implant Tablets, emulsified injections, implants and pharmaceutically viable long-acting controlled release dosage forms.