TWI554272B - Pharmaceutical composition comprising acarbose and its use for immunomodulatory - Google Patents

Description

包含阿卡波糖之醫藥組合物及其用於免疫調節之用途 Pharmaceutical composition containing acarbose and its use for immunomodulation

本發明關於一包含阿卡波糖的新用途，特別是阿卡波糖用於調節免疫或治療免疫發炎病症的新用途。 The present invention relates to a novel use comprising acarbose, in particular for the use of acarbose for the modulation of immunity or for the treatment of immunoinflammatory conditions.

免疫發炎之特徵為身體的免疫反應被不當地活化。免疫系統不對感染外來物加以攻擊，而對身體本身的組織或移植組織加以攻擊與損害。被免疫系統攻擊之組織因疾病有所不同。例如，於多發性硬化症，免疫反應係對抗神經組織，而克隆氏症，則對抗消化道。免疫炎症疾病侵擾數百萬人，且包括的病症例如有氣喘、過敏性眼內炎症、關節炎、異位性皮膚炎、異位性濕疹、糖尿病、溶血性貧血、發炎性皮膚炎、發炎性腸道或胃腸病症(例如，克隆氏症及潰瘍性大腸炎)、多發性硬化症、重症肌無力、搔癢症/發炎、牛皮癬、類風濕性關節炎、肝硬化及全身性紅斑狼瘡。 Immune inflammation is characterized by the body's immune response being improperly activated. The immune system does not attack infected foreign objects, but attacks and damages the body's own tissues or transplanted tissues. Tissues attacked by the immune system vary from disease to disease. For example, in multiple sclerosis, the immune response is against nerve tissue, while Crohn's disease is against the digestive tract. Immune inflammatory diseases inflict millions of people, including diseases such as asthma, allergic intraocular inflammation, arthritis, atopic dermatitis, atopic eczema, diabetes, hemolytic anemia, inflammatory dermatitis, inflammation Sexual intestinal or gastrointestinal disorders (eg, Crohn's disease and ulcerative colitis), multiple sclerosis, myasthenia gravis, pruritus/inflammation, psoriasis, rheumatoid arthritis, cirrhosis, and systemic lupus erythematosus.

目前對免疫發炎疾病已有多種藥物開發，但該等藥劑之效果不一且常會伴隨有不佳的副作用。故，需要有對於免疫炎症疾病治療更好的治療藥劑及方法。 A variety of drugs have been developed for immunoinflammatory diseases, but the effects of these agents are different and often accompanied by poor side effects. Therefore, there is a need for better therapeutic agents and methods for the treatment of immune inflammatory diseases.

鑒於上述問題，本案發明人意外的發現，已知用於治療第二 型糖尿病的藥物，阿卡波糖，其係一種葡萄糖苷酶抑制劑類藥物，與小腸絨毛上的α-葡萄糖苷酶作可逆性的競爭抑制，以減緩醣類在小腸內的分解，因此，可延遲醣類的吸收，以避免飯後高血糖值的現象，而達到較平穩的飯後血糖控制；經本發明之實驗後發現具有調節免疫的功能，而能夠用於治療免疫發炎病症。 In view of the above problems, the inventors of the present invention unexpectedly discovered that it is known for treating the second Type 2 diabetes drug, acarbose, which is a glucosidase inhibitor, reversibly competitively inhibits α-glucosidase on the intestinal villi to slow the breakdown of sugars in the small intestine, therefore, The absorption of sugar can be delayed to avoid the phenomenon of post-prandial high blood sugar level, and a smoother postprandial blood glucose control can be achieved; after the experiment of the present invention, it is found to have the function of regulating immunity, and can be used for treating immune inflammatory diseases.

因此，本發明一方面係提供一種用於治療免疫發炎病症之醫藥組合物，其包含阿卡波糖(acarbose)。 Accordingly, in one aspect, the invention provides a pharmaceutical composition for treating an immunoinflammatory condition comprising acarbose.

在本發明之一實施例中，該免疫發炎病症為：類風濕性關節炎、乾癬、第一型糖尿病、成人性史底耳氏症、視神經脊髓炎、克隆氏症、潰瘍性大腸炎、氣喘、慢性阻塞性肺病、風濕性多肌痛症、巨細胞動脈炎、全身性紅斑狼瘡、異位性皮膚炎、多發性硬化症、重症肌無力、牛皮癬、僵直性脊椎炎、肝硬化或乾癬性關節炎。 In one embodiment of the present invention, the immune inflammatory condition is: rheumatoid arthritis, cognac, type 1 diabetes, adult history of vertebral disease, optic neuromyelitis, Crohn's disease, ulcerative colitis, asthma , chronic obstructive pulmonary disease, rheumatic polymyalgia, giant cell arteritis, systemic lupus erythematosus, atopic dermatitis, multiple sclerosis, myasthenia gravis, psoriasis, ankylosing spondylitis, cirrhosis or dryness arthritis.

於本發明之一較佳實施例中，本發明之醫藥組合物可進一步包含一第二藥物；其中該第二藥物較佳係選自由非類固醇抗發炎藥(NSAID)、COX-2抑制劑、生物分子、免疫調節劑、小分子免疫調節劑、疾病修飾抗風濕藥(DMARD)、黃嘌呤、抗膽鹼化合物、β受體協同劑、支氣管擴張劑、非類固醇免疫親和蛋白依存免疫抑制劑、維生素D類似物、補骨脂素、維生素A酸及5-胺基水楊酸所構成之群。於一更佳實施例中，該疾病修飾抗風濕藥(DMARD)為托法替尼(tofactinib)。 In a preferred embodiment of the present invention, the pharmaceutical composition of the present invention may further comprise a second drug; wherein the second drug is preferably selected from the group consisting of a non-steroidal anti-inflammatory drug (NSAID), a COX-2 inhibitor, Biomolecules, immunomodulators, small molecule immunomodulators, disease modifying antirheumatic drugs (DMARD), jaundice, anticholinergic compounds, beta receptor synergists, bronchodilators, nonsteroidal immunoaffinity protein-dependent immunosuppressants, A group consisting of a vitamin D analog, psoralen, vitamin A acid, and 5-aminosalicylic acid. In a more preferred embodiment, the disease modifying antirheumatic drug (DMARD) is tofactinib.

於本發明之實施例中，該醫藥組合物可為任何劑型，較佳係呈錠劑或膠囊之型式。於另一較佳實施例中，該醫藥組合物配方為局部投予用藥或全身投予用藥。 In an embodiment of the invention, the pharmaceutical composition may be in any dosage form, preferably in the form of a lozenge or capsule. In another preferred embodiment, the pharmaceutical composition is formulated for topical administration or systemic administration.

本發明之另一方面係一種阿卡波糖供製備免疫調節藥物之用途。 Another aspect of the invention is the use of acarbose for the preparation of an immunomodulatory drug.

非限制且非詳盡之實施例將結合附圖說明如下。必須了解的是下列圖式僅依據本說明書公開之內容描繪數個實施例，因此，勿因而限制其範圍，本發明所揭示之內容將特別詳細的藉由這些圖式作為說明，其中：圖1顯示CIA小鼠實驗組與對照組中，A：CIA發生率比較圖；B：患病腳掌數量比較圖；C：腳掌照片圖；D：關節炎評分比較圖；E：踝關節的組織學(H&E)照片；F：對發炎細胞的浸潤、滑膜增生以及腳踝關節的骨侵蝕之組織學比較圖。 Non-limiting and non-exhaustive embodiments are described below in conjunction with the drawings. It is to be understood that the following drawings depict only a few embodiments in accordance with the disclosure of the present disclosure, and therefore, the scope of the present invention is not to be construed as limited. A comparison of the incidence of A:CIA in the CIA mice experimental group and the control group; B: comparison of the number of diseased feet; C: photo of the sole of the foot; D: comparison of the arthritis score; E: histology of the ankle joint ( H&E) photograph; F: histological comparison of infiltration of inflamed cells, synovial hyperplasia, and bone erosion of the ankle joint.

圖2顯示CIA小鼠實驗組與對照組中，以ELISA測定A：腳掌組織；及B：血清中選擇性細胞激素的含量。 Figure 2 shows the content of selective cytokines in A: foot tissue; and B: serum in CIA mice experimental group and control group.

圖3顯示CIA小鼠實驗組與對照組中，A：抗CII IgG比較圖；B：CII抗原刺激增生比較圖；C：淋巴球細胞上清液的不同細胞激素含量比較圖。 Figure 3 shows a comparison of A: anti-CII IgG in the CIA mouse experimental group and the control group; B: comparison of CII antigen-stimulated hyperplasia; C: comparison of different cytokine contents in the lymphocyte supernatant.

圖4顯示不同劑量及不同藥劑對CIA小鼠之功效評估比較圖；A：托法替尼1mg/kg/day、5mg/kg/day、25mg/kg/day劑量；B：托法替尼1mg/kg/day、阿卡波糖500mg/kg/day、阿卡波糖500mg/kg/day合併托法替尼1mg/kg/day；C：托法替尼5mg/kg/day、阿卡波糖500mg/kg/day、阿卡波糖500mg/kg/day合併托法替尼5mg/kg/day。 Figure 4 shows a comparison of the efficacy of different doses and different agents in CIA mice; A: tofartinib 1mg/kg/day, 5mg/kg/day, 25mg/kg/day dose; B: tofacitinib 1mg /kg/day, acarbose 500mg/kg/day, acarbose 500mg/kg/day combined with tofartinib 1mg/kg/day; C: tofartinib 5mg/kg/day, Akapo Sugar 500mg/kg/day, acarbose 500mg/kg/day combined with tofartinib 5mg/kg/day.

圖5顯示以咪喹莫特62.5mg/kg/day誘發小鼠乾癬佐以阿卡波 糖500mg/kg/day治療的實驗組與對照組之A：皮膚照片圖；B：臨床表徵評分比較圖。 Figure 5 shows that imiquimod 62.5 mg / kg / day induced mouse cognac with Akapo A: skin photo of the experimental group and the control group treated with sugar 500 mg/kg/day; B: comparison chart of clinical characterization scores.

圖6顯示乾癬小鼠皮膚組織學(H&E)照片，A：控制組乳膏； B：咪喹莫特誘發乾癬；C：以咪喹莫特誘發乾癬佐以阿卡波糖50mg/kg/day治療；D：以咪喹莫特誘發乾癬阿卡波糖50mg/kg/day治療。 Figure 6 shows the skin histology (H&E) of dried mice, A: control group cream; B: imiquimod induced dryness; C: imiquimod-induced dryness with acarbose 50 mg/kg/day; D: imiquimod-induced dry acarbose 50 mg/kg/day .

於下文中，將對本發明所揭示之示例性實施例及範例進行詳細的描述，並且附有圖式，使得本發明之概念得以經由本發明技術領域人員輕易實行。 In the following, the exemplary embodiments and examples disclosed herein are described in detail, and the drawings are attached to the drawings, and the concept of the present invention can be easily implemented by those skilled in the art.

然而，必須注意的是本發明所揭示之內容並不應被限制於發明實施例，且可以不同方式實現。於圖式中，部份與本發明不直接相關之內容省略以提高圖式之清晰度，標號於文件當中標示。 However, it must be noted that the disclosure of the present invention should not be limited to the embodiments of the invention and may be implemented in different ways. In the drawings, parts that are not directly related to the present invention are omitted to improve the clarity of the drawings, and the labels are indicated in the documents.

整篇說明書當中，所使用之詞彙「包含(comprises)」或「包括(includes)」意謂著除了描述的組成、步驟、操作指令及/或元素以外，不排除一或多個其他組成、步驟、操作指令及/或存在或附加元素。所使用之詞彙「大約(about)或約(approximately)」意指具有接近或可允許的誤差範圍，用於避免本發明所揭示之準確或絕對的數值受未知的第三方非法或非正當使用的。詞彙「一」意指該冠詞之語法對象為一或一個以上(例如：至少為一)。 The word "comprises" or "includes" as used throughout the specification does not exclude one or more other components or steps other than the described components, steps, operation instructions and/or elements. , operating instructions and/or existing or additional elements. The word "about" or "approximately" is used to mean that there is a near or permissible range of error for avoiding the use of the precise or absolute value disclosed herein by an unknown third party. . The term "a" means that the grammatical object of the article is one or more (eg, at least one).

本發明係提供一種用於治療免疫發炎病症之醫藥組合物，其包含阿卡波糖(acarbose)，以及一種阿卡波糖供製備免疫調節藥物之用途。 The present invention provides a pharmaceutical composition for treating an immunoinflammatory condition comprising acarbose and an acarbose for use in the preparation of an immunomodulatory drug.

本文中術語「治療」意指投予或處方一組成物至一患者以治 療或預防免疫發炎病症。本文中「患者」意指任何動物(例如，人類)。其他可用本發明之方法、組成物及套組治療之動物包括馬、狗、貓、豬、山羊、兔、倉鼠、猴、天竺鼠、大鼠、小鼠、蜥蜴、蛇、綿羊、牛、魚及鳥。 The term "treatment" as used herein means administering or prescribing a composition to a patient. Treat or prevent immune inflammatory conditions. By "patient" is meant herein any animal (eg, human). Other animals which can be treated by the methods, compositions and kits of the invention include horses, dogs, cats, pigs, goats, rabbits, hamsters, monkeys, guinea pigs, rats, mice, lizards, snakes, sheep, cattle, fish and bird.

本文中術語「免疫發炎病症」包括許多情況，包括但不限於 自體免疫疾病、增生性皮膚病及發炎性濕疹。免疫發炎病症會導致身體的健康組織被發炎過程破壞、使免疫系統失去控制及造成不欲之細胞增殖。 免疫發炎病症之例如尋常痤瘡；急性呼吸困難症候群；艾迪生氏症(Addison’s病症)；過敏性鼻炎；過敏性眼內發炎病症、ANCA-相關性小-血管血管炎；僵直性脊椎炎；關節炎、氣喘；動脈硬化症；異位性皮膚炎；自體免疫溶血性貧血；自體免疫肝炎；貝西氏(Behcet’s)病；貝爾氏痲痹(Bell’s palsy)；大水皰性類天皰瘡(bullous pemphigoid)；腦缺血；慢性阻塞性肺病；肝硬化；柯剛氏(Cogan’s)症候群；接觸性皮膚病；COPD；克隆氏病；庫欣氏(Cushing’s)症候群；皮膚肌炎；糖尿病；圓型紅斑狼瘡；嗜伊紅性肌膜炎；紅斑性結節；剝落性皮膚炎；纖維肌痛；腎絲球硬化症；巨細胞動脈炎；痛風；痛風性關節炎；移殖物抗寄主疾病；手部濕疹；過敏性紫斑症(Henoch-Schonlein purpura)；妊娠皰疹性(herpes gestationis)；多毛症；特發性角-鞏膜炎；特發性肺纖維化；特發性血小板減少性紫斑；發炎性腸或胃腸疾病、發炎皮膚病；扁平苔蘚；狼瘡性腎炎；淋巴瘤性氣管支氣管炎；黃斑部水腫；多發性硬化症；重症肌無力；肌炎；骨性關節炎；胰臟炎；天瘡樣疹症；尋常天瘡；多發性結節性動脈炎；風濕性多肌痛；陰囊搔癢症；搔癢症/發炎；牛皮癬；乾癬性關節炎；類風濕性關節炎；再發性的多發性軟骨炎；類肉瘤病引起之玫瑰斑；硬皮病引起之玫瑰斑；史維特氏 (Sweet’s)症候群造成之玫瑰斑；全身性紅斑狼瘡造成之玫瑰斑；蕁麻疹引起之玫瑰斑；帶疹相關疼痛引起之玫瑰斑；類肉瘤病；硬皮病；局部腎絲球硬化症；敗血性休克症候群；肩肌腱炎或滑囊炎；修格連氏(Sjogren’s)症候群；史底耳氏(Still’s)症；中風誘發性腦細胞死亡；史維特氏病；全身性紅斑狼瘡；全身性硬化症；高安氏(Takayasu’s)動脈炎；顳動脈炎；毒性皮膚壞死；肺結核；第I型糖尿病；潰瘍性大腸炎；葡萄膜炎；血管炎；及維格內氏(Wegener’s)囊腫。於本案之較佳實施例中，該免疫發炎病症包含類風濕性關節炎、乾癬、第一型糖尿病、成人性史底耳氏症、視神經脊髓炎、克隆氏症、潰瘍性大腸炎、氣喘、慢性阻塞性肺病、風濕性多肌痛症、巨細胞動脈炎、全身性紅斑狼瘡、異位性皮膚炎、多發性硬化症、重症肌無力、牛皮癬、僵直性脊椎炎、肝硬化或乾癬性關節炎或其他與細胞激素IL-17失調的相關免疫發炎病症。 The term "immune inflammatory condition" as used herein includes many situations, including but not limited to Autoimmune diseases, proliferative skin diseases and inflammatory eczema. Immune inflammatory conditions can cause the body's healthy tissues to be destroyed by the inflammatory process, the immune system to lose control, and the unwanted cells to proliferate. Immunoinflammatory conditions such as acne vulgaris; acute dyspnea syndrome; Addison's disease; allergic rhinitis; allergic intraocular inflammation, ANCA-related small-vascular vasculitis; ankylosing spondylitis; arthritis , asthma; atherosclerosis; atopic dermatitis; autoimmune hemolytic anemia; autoimmune hepatitis; Behcet's disease; Bell's palsy; large vesicular pemphigus (bullous) Pemphigoid); cerebral ischemia; chronic obstructive pulmonary disease; cirrhosis; Cogan's syndrome; contact dermatosis; COPD; Crohn's disease; Cushing's syndrome; dermatomyositis; Lupus erythematosus; eosinophilic fasciitis; erythematous nodules; exfoliative dermatitis; fibromyalgia; glomerular sclerosis; giant cell arteritis; gout; gouty arthritis; Eczema; Henoch-Schonlein purpura; Herpes gestationis; hirsutism; idiopathic horn-scleraitis; idiopathic pulmonary fibrosis; idiopathic thrombocytopenia Purpura; inflammatory bowel or gastrointestinal disease, inflammatory skin disease; lichen planus; lupus nephritis; lymphoma tracheitis; macular edema; multiple sclerosis; myasthenia gravis; myositis; osteoarthritis; Dampitis; acne-like rash; common sore; multiple nodular arteritis; rheumatic polymyalgia; scrotal pruritus; pruritus/inflammation; psoriasis; dry arthritis; rheumatoid arthritis; Sexual polychondritis; rose plaque caused by sarcoma-like disease; rose plaque caused by scleroderma; (Sweet's) syndrome caused by rose spots; rose spots caused by systemic lupus erythematosus; rose plaque caused by urticaria; rose plaque caused by rash-related pain; sarcoma-like disease; scleroderma; local glomerulosclerosis; Hemorrhagic shock syndrome; scapular tendonitis or bursitis; Sjogren's syndrome; Still's disease; stroke-induced brain cell death; Schwitter's disease; systemic lupus erythematosus; systemic sclerosis Disease; Takayasu's arteritis; temporal arteritis; toxic skin necrosis; tuberculosis; type I diabetes; ulcerative colitis; uveitis; vasculitis; and Wegener's cyst. In a preferred embodiment of the present invention, the immunoinflammatory condition comprises rheumatoid arthritis, cognac, type 1 diabetes, adult sexual history, optic neuromyelitis, Crohn's disease, ulcerative colitis, asthma, Chronic obstructive pulmonary disease, rheumatic polymyalgia, giant cell arteritis, systemic lupus erythematosus, atopic dermatitis, multiple sclerosis, myasthenia gravis, psoriasis, ankylosing spondylitis, cirrhosis or dry joints Inflammation or other immunoinflammatory condition associated with dysregulation of the cytokine IL-17.

本文中術語「非皮膚發炎病症」包括，例如類風濕性關節炎、 發炎腸病、氣喘及慢性阻塞性肺病。 The term "non-skin inflammatory condition" as used herein includes, for example, rheumatoid arthritis, Inflammatory bowel disease, asthma and chronic obstructive pulmonary disease.

本文中術語「皮膚發炎病症」或「發炎性皮膚炎」包括，例 如牛皮癬、急性熱嗜中性皮膚病、濕疹(例如缺脂性濕疹、汗疹、小囊掌跖處濕疹)、漿細胞性局限性陰莖頭炎、陰莖頭***炎、貝西氏病(Behcet’s病症)、離心性環形紅斑、持久性色素障礙性紅斑、多形紅斑、環狀肉芽腫、光澤苔蘚、扁平苔蘚、硬化萎縮性苔癬、慢性單純苔癬、小棘苔蘚、錢串狀皮膚炎、壞疽性膿皮病、類肉瘤病、角膜下膿皰病、蕁麻疹及過渡性棘皮溼疹。「增生性皮膚病」意指良性或惡性病其特徵為上皮或表皮之細胞***加速。增生性皮膚病之例子有牛皮癬、異位性皮膚炎、非特異性皮膚炎、 初級刺激物接觸皮膚炎、過敏性接觸皮膚炎、基底及鱗狀細胞皮膚癌、層狀鱗癬、表皮鬆懈角化過度症、前惡性角化病、粉刺及脂溢性皮膚炎。熟悉該領域技術者應可了解，某特定疾病、病症或情形可同時定性為增生性皮膚病及發炎濕疹。該疾病之例為牛皮癬。 The term "skin inflammatory condition" or "inflammatory dermatitis" as used herein includes, for example, Such as psoriasis, acute heat neutrophil skin disease, eczema (such as fat-free eczema, sweat rash, eczema at the small cystic sac), plasma cell-limited penile head inflammation, penile head dermatitis, Bezi's disease ( Behcet's condition), eccentric ring erythema, persistent pigmented erythema, polymorphous erythema, ring granuloma, lustrous moss, lichen planus, hardened atrophic moss, chronic simple moss, small spine moss, money-like skin Inflammation, gangrenous pyoderma, sarcoma, subcorneal impetigo, urticaria and transitional echinodermitis. "Proliferative skin disease" means a benign or malignant disease characterized by accelerated cell division of the epithelium or epidermis. Examples of proliferative skin diseases are psoriasis, atopic dermatitis, non-specific dermatitis, Primary irritants are exposed to dermatitis, allergic contact with dermatitis, basal and squamous cell skin cancer, lamellar lemma, epidermal hyperkeratosis, anterior malignant keratosis, acne and seborrheic dermatitis. Those skilled in the art will appreciate that a particular disease, condition or condition can be characterized as both proliferative skin disease and inflammatory eczema. An example of this disease is psoriasis.

本發明之醫藥組合物可進一步包含一第二藥物。其中，該第二藥物較佳係選自由非類固醇抗發炎藥(NSAID)、COX-2抑制劑、生物分子、免疫調節劑、小分子免疫調節劑、疾病修飾抗風濕藥(DMARD)、黃嘌呤、抗膽鹼化合物、β受體協同劑、支氣管擴張劑、非類固醇免疫親和蛋白依存免疫抑制劑、維生素D類似物、補骨脂素、維生素A酸及5-胺基水楊酸所構成之群。 The pharmaceutical composition of the present invention may further comprise a second drug. Wherein, the second drug is preferably selected from the group consisting of a non-steroidal anti-inflammatory drug (NSAID), a COX-2 inhibitor, a biomolecule, an immunomodulator, a small molecule immunomodulator, a disease modifying antirheumatic drug (DMARD), and jaundice. , an anticholinergic compound, a beta receptor synergist, a bronchodilator, a non-steroidal immunoaffinity protein-dependent immunosuppressive agent, a vitamin D analog, a psoralen, a vitamin A acid, and a 5-aminosalicylic acid group.

本文中術語「非類固醇抗發炎藥」包含但不限於奈普生鈉、雙氯芬酸鈉、雙氯芬酸鉀、阿司匹靈、舒林酸、雙氟尼酸、吡羅昔康、引朵美辛、布洛芬、萘丁美酮、膽鹼三水楊酸鎂、水楊酸鈉、水楊酸基水楊酸(三柳膽鎂，salsalate)、苯氧布洛芬、氟比洛芬、酮洛芬、甲氯芬那酸鈉、美洛昔康、奧沙普嗪、舒林酸及托美汀。 The term "non-steroidal anti-inflammatory drugs" as used herein includes, but is not limited to, sodium naproxen, diclofenac sodium, diclofenac potassium, aspirin, sulindac, diflunisal, piroxicam, indomethacin, cloth Lofen, nabumetone, choline magnesium salicylate, sodium salicylate, salicylic acid salicylic acid (sallium), phenoxyprofen, flurbiprofen, ketoprofen , meclofenac sodium, meloxicam, oxaprozin, sulindac and tolmetine.

本文中術語「疾病修飾抗風濕藥」包含但不限於滅殺除癌(Methotrexate)、硫唑嘌呤(Azathioprine)、來氟米特(Leflunomide)、柳氮磺胺吡啶(Sulfasalazine)、羥氯奎(Hydroxychloroquine)、托法替尼(Tofactinib)。 The term "disease-modifying antirheumatic drug" as used herein includes, but is not limited to, Methotrexate, Azathioprine, Leflunomide, Sulfasalazine, Hydroxychloroquine. ), Tofatinib (Tofactinib).

本文中術語「免疫調節劑」包含但不限於環孢靈素(Cyclosporin)、咪喹莫特(imiquimod)。 The term "immunomodulator" as used herein includes, but is not limited to, cyclosporin, imiquimod.

本文中術語「抗組織胺」意指一化合物其阻斷組織胺之作用。抗組織胺類型包括，但不限定於，乙醇胺、乙二胺、吩噻嗪、烷基胺、 哌嗪及哌啶。 The term "antihistamine" as used herein means a compound which blocks the action of histamine. Antihistamine types include, but are not limited to, ethanolamine, ethylenediamine, phenothiazine, alkylamine, Piperazine and piperidine.

本文中術語「非類固醇免疫親和蛋白依存性免疫抑制劑」或「NsIDI」意指任何非類固醇藥劑其可減低原發炎細胞激素生產或分泌、結合免疫親和蛋白或造成原發炎反應之向下調控。NsIDI包括鈣調神經磷解酶(calcineurin)抑制劑，例如環孢素、肝磷脂(tacrolimus)、子囊黴素、美莫司(pimecrolimus)及其他可抑制鈣調神經磷解酶之磷解酶活性的藥劑(肽、肽片段、化學性修飾肽或肽擬似物擬似物)。NsIDI亦包括雷帕黴素(rapamycin)(西羅莫司，sirolimus)及依維莫司(everolimus)，其會結合於FK506-結合蛋白質、FKBP-12，並阻斷白血球之抗原誘發性增生以及細胞激素分泌。 The term "non-steroidal immunoaffinity protein-dependent immunosuppressive agent" or "NsIDI" as used herein means any non-steroidal agent which reduces the production or secretion of primary inflammatory cytokines, binds to immunoaffinity proteins or causes down-regulation of the primary inflammatory response. NsIDI includes calcium calcineurin inhibitors such as cyclosporine, heparin (licrolimus), ascomycin, pimecrolimus and other phospholyase activities that inhibit calcineurin Agent (peptide, peptide fragment, chemically modified peptide or peptide mimetic). NsIDI also includes rapamycin (sirolimus) and everolimus, which bind to FK506-binding protein, FKBP-12, and block antigen-induced proliferation of leukocytes and Cytokine secretion.

本文中術語「小分子免疫調節劑」意指非類固醇、非-NsIDI化合物其可減少原發炎細胞激素生產或分泌，導致原發炎反應之向下調控，或者以免疫親和蛋白-非依存的方式調節免疫系統。小分子免疫調節劑之例子有p38 MAP激酶抑制劑，例如VX 702(Vertex Pharmaceuticals)、SCIO 469(Scios)、多拉馬皮莫(doramapimod)(Boehringer Ingelheim)、RO 30201195(Roche)及SCIO 323(Scios)，TACE抑制劑例如有DPC 333(Bristol Myers Squibb)、ICE抑制劑例如有普納卡桑(pranalcasan)(Vertex Pharmaceuticals)，IMPDH抑制劑例如黴酚酸酯(mycophenolate)(Roche)及梅利媒珀地補(merimepodib)(Vertex Pharamceuticals)。 The term "small molecule immunomodulator" as used herein means a non-steroidal, non-NsIDI compound that reduces the production or secretion of primary inflammatory cytokines, leading to downregulation of the primary inflammatory response, or by immunoaffinity protein-independent manner. immune system. Examples of small molecule immunomodulators are p38 MAP kinase inhibitors such as VX 702 (Vertex Pharmaceuticals), SCIO 469 (Scios), doramapimod (Boehringer Ingelheim), RO 30201195 (Roche) and SCIO 323 ( Scios), TACE inhibitors such as DPC 333 (Bristol Myers Squibb), ICE inhibitors such as pranalcasan (Vertex Pharmaceuticals), IMPDH inhibitors such as mycophenolate (Roche) and Meli Merimepodib (Vertex Pharamceuticals).

本發明之醫藥組合物之劑量可依照技藝人士針對患者的狀況做調整，提供足量的活性成份，或可選用低劑量、中劑量或高劑量。投予的劑量效果可由熟練的醫師使用任何適於既定徵兆之標準方法判斷。 The dosage of the pharmaceutical composition of the present invention can be adjusted to the condition of the patient according to the skilled person, providing a sufficient amount of the active ingredient, or alternatively a low dose, a medium dose or a high dose. The dose effect administered can be judged by a skilled physician using any standard method suitable for the intended indication.

本文中術語「低劑量」意指較特定化合物(以既定路徑投予 以治療任何人類病症)之最低標準建議劑量低至少5%(例如，至少10%、20%、50%、80%、90%或甚至95%)例如，以吸入投予之皮質類固醇其低劑量與以口服投予之皮質類固醇其低劑量不同。 The term "low dose" as used herein refers to a specific compound (administered in a defined path) The minimum standard for treating any human condition is recommended to be at least 5% lower (eg, at least 10%, 20%, 50%, 80%, 90%, or even 95%), eg, a low dose of a corticosteroid administered by inhalation. It is different from the low dose of corticosteroid administered orally.

本文中術語「高劑量」意指較特定化合物(用以治療任何人 類病症)之最高標準建議劑量高至少5%(例如，至少10%、20%、50%、100%、200%或甚至300%)。「中劑量」意指劑量介於低劑量與高劑量之間。 The term "high dose" as used herein means a specific compound (for treating any person) The highest standard for a class of conditions) suggests a dose of at least 5% (eg, at least 10%, 20%, 50%, 100%, 200%, or even 300%). "Medium dose" means that the dose is between a low dose and a high dose.

本文中術語「足量」意指本發明中化合物之組合量其係以臨 床上相關方式治療或預防免疫炎症時所須之量。用於實施本發明以治療由於免疫炎症疾病之活性化合物其足夠量依投予方式、年齡、體重及患者的一般健康情形有所不同。 The term "sufficient amount" as used herein means the combination of the compounds of the present invention. The amount of bed needed to treat or prevent immune inflammation. The amount of active compound used to practice the present invention to treat an immune inflammatory disease varies depending on the mode of administration, age, weight, and general health of the patient.

本發明之醫藥組合物具體例之投予路徑包括，但不限定於局 部投予、經皮及全身性投予(例如，靜脈內、肌肉內、吸入、直腸內、領、***、腹膜內、關節內、眼或口服投予)。任何前述投予路徑可與傳統對治療免疫發炎病症之藥物一同投予。 The administration route of the specific example of the pharmaceutical composition of the present invention includes, but is not limited to, a bureau Partial, percutaneous, and systemic (eg, intravenous, intramuscular, inhalation, rectal, collar, vaginal, intraperitoneal, intra-articular, ocular, or oral). Any of the aforementioned routes of administration can be administered in conjunction with conventional drugs for treating immunoinflammatory conditions.

本發明之醫藥組合物可以為任何種型式，例如錠劑、膠囊、 藥丸、粉末、顆粒、懸浮劑、乳化劑、溶液、凝膠(包括水凝膠)、糊劑、油膏、乳霜、膏藥、浸劑、滲透運送工具、栓劑、灌腸劑、注射劑、植入劑、噴霧或氣溶膠。該等組成物可依傳統製藥規範配製，其中較佳為口服劑型，如錠片、膠囊、或糖漿；或為直腸投予，如栓劑；或為其他種劑型，如持續釋放或緩釋劑型等。 The pharmaceutical composition of the present invention may be in any type, such as tablets, capsules, Pills, powders, granules, suspensions, emulsifiers, solutions, gels (including hydrogels), pastes, ointments, creams, plasters, infusions, osmotic delivery vehicles, suppositories, enemas, injections, implants Agent, spray or aerosol. Such compositions may be formulated according to conventional pharmaceutical specifications, preferably oral dosage forms such as tablets, capsules, or syrups; or rectal administration, such as suppositories; or other dosage forms, such as sustained release or sustained release dosage forms, and the like. .

本發明之醫藥組合物中，如含有第二藥物，則各化合物可用 該技術領域已知的方法配方。例如，第一及第二藥劑可以同時或分別的配方。較佳為，第一及第二藥劑同時一起投予或接近同時投予；若同時投予，則可一起配方於同一藥丸、膠囊、液劑等。藉由對不同藥劑使用不同的配方策略，可以適當配合各藥劑的藥物動力學曲線。個別或分別配方的藥劑可包在一起成為一個套組。無限定的套組實施例，例如包含2個藥丸、一個藥丸及粉末、栓劑及一藥瓶內液劑、2種外用乳霜等形式。該套組可包含選擇性的配件，其會幫助對患者投予單位劑量，成分包括例如用於泡製粉末型之藥瓶、注射針筒、客製IV運送系統、吸入劑等。此外，該單位劑量套組可包括製備及投予該組成物的說明。該套組可製造為一位患者使用的單一劑量、對特定患者之多次使用劑量(可為相同劑量或者個別化合物隨治療進展其效力不同)；或該套組可包括適用於對多位患者投予之多次劑量(“大包裝”)。該套組可組合在紙盒、泡形罩包裝、瓶子、管子等。或者，可調配成控制型及/或持續釋放，該「持續釋放」或「控制性釋放」意指治療上活性成分係從配方中以受控制的速度釋出，以使該成分可長時間(例如，約12~24小時)維持治療上有效的血液水平(低於有毒性之水平)，故可製成12小時或24小時之劑型。 In the pharmaceutical composition of the present invention, if a second drug is contained, each compound can be used. Method formulations known in the art. For example, the first and second medicaments can be formulated simultaneously or separately. Preferably, the first and second medicaments are administered simultaneously or nearly simultaneously; if administered simultaneously, they may be formulated together in the same pill, capsule, liquid, and the like. By using different formulation strategies for different agents, the pharmacokinetic profile of each agent can be appropriately matched. Individual or separately formulated medicaments can be packaged together into a single kit. An unrestricted kit embodiment includes, for example, two pills, one pill and powder, a suppository and a vial solution, and two external creams. The kit may include optional accessories that will aid in the administration of a unit dose to a patient, including, for example, vials for infusion of powders, syringes, custom IV delivery systems, inhalants, and the like. Additionally, the unit dose kit can include instructions for preparing and administering the composition. The kit can be manufactured as a single dose for one patient, multiple doses for a particular patient (which can be the same dose or the effectiveness of individual compounds as the treatment progresses); or the kit can include multiple patients Multiple doses administered ("big package"). The kit can be combined in a carton, blister pack, bottle, tube, and the like. Alternatively, it may be formulated to be controlled and/or sustained release, and the "sustained release" or "controlled release" means that the therapeutically active ingredient is released from the formulation at a controlled rate such that the ingredient is prolonged ( For example, about 12 to 24 hours) maintains a therapeutically effective blood level (below a toxic level) so that it can be made into a 12 hour or 24 hour dosage form.

[實施例][Examples]

在下文中，將利用範例及圖式特別描寫本發明所揭示之內容。然而，本發明所揭示之內容不限制於該範例及圖式。 In the following, the disclosure of the present invention will be specifically described using examples and drawings. However, the disclosure of the present invention is not limited to this example and the drawings.

實施例1-類風濕性關節炎與阿卡波糖關係的統計學調查Example 1 - Statistical investigation of the relationship between rheumatoid arthritis and acarbose

[以族群作為基礎案例對照研究] [Based on ethnic group as a case-control study]

數據源為使用來自台灣的國民健康保險研究數據庫 (NHIRD)1999-2011以人群做為基礎，匹配的病例對照研究。NHIRD為超過98%的台灣人口全面的加密醫療數據，而為研究目的發布。該NHIRD於2000年隨機選取百萬參保者，形成具有代表性的數據庫，該數據庫中包含潛在致殘性自身免疫性疾病，包括類風濕性關節炎(RA)。RA的診斷再經由美國風濕病醫學會RA分類標準(1987)進行驗證。 The data source is the National Health Insurance Research Database from Taiwan. (NHIRD) 1999-2011 Population-based, matched case-control study. NHIRD is a comprehensive encrypted medical data for more than 98% of the Taiwanese population, and is released for research purposes. The NHIRD randomly selected millions of participants in 2000 to form a representative database containing potentially disabling autoimmune diseases, including rheumatoid arthritis (RA). The diagnosis of RA was further verified by the American College of Rheumatology RA Classification Standard (1987).

[研究樣本] [research sample]

糖尿病(DM)案例 Diabetes (DM) case

病患初步診斷為糖尿病日[國際分類疾病，第九次修訂，臨 床修改(ICD9-CM)代碼250.x]在2000年1月1日之後，並同時處方任何抗糖尿病藥物28天者被列為DM病患。DM診斷日期用於測量DM病程時間的係定義為初始抗糖尿病藥物處方的時間。 The patient was initially diagnosed with Diabetes Day [International Classification Disease, Ninth Revision, Clinical Revision (ICD9-CM) Code 250.x] after January 1, 2000, and prescribes any antidiabetic drug at the same time. 28 days were classified as DM patients. The DM diagnostic date is used to measure the duration of the DM course as the time to prescribe the initial antidiabetic drug.

類風濕性關節炎(RA)案例 Rheumatoid arthritis (RA) case

確定在2001至2010年間有24,429位具有重大傷病證(RA診斷 時年齡16)新診斷為RA的病患，為了最大限度地減少反因果關係的可能性，排除了在開始抗糖尿病治療一年內患有RA的患者。在這些RA患者中，確定了723位病患為第一次診斷(指標日期)為RA之日前具有至少一年的糖尿病史。 It is determined that there are 24,429 major injuries and illnesses between 2001 and 2010 (the age at diagnosis of RA) 16) Patients with a new diagnosis of RA, in order to minimize the possibility of anti-causal relationships, exclude patients with RA within one year of starting anti-diabetic treatment. Among these RA patients, 723 patients were identified as having a history of diabetes for at least one year prior to the date of the first diagnosis (index date) of RA.

非類風濕性關節炎(non-RA)控制組 Non-rheumatoid arthritis (non-RA) control group

使用KHID20000，匹配RA案例的RA診斷年齡(16-25、 26-35、36-45、46-55、56-65或>65歲)、性別以及指標日年份(指標年份)，並在1999-2011年份中隨機選取7,230位之DM患者，其患有DM病史超過一年且從未診斷出RA作為非RA控制組。選用第一次門診時間作為控制組的指標 日期。 Use KHID20000 to match the RA diagnosis age of RA cases (16-25, 26-35, 36-45, 46-55, 56-65 or >65 years old), gender and indicator date year (indicator year), and randomly selected 7,230 DM patients with DM in 1999-2011 year The medical history was more than one year and RA was never diagnosed as a non-RA control group. Use the first outpatient time as an indicator of the control group date.

[使用抗糖尿病藥物] [Use anti-diabetes drugs]

基於NHIRD的處方紀錄，在指標日之前，一年內有28天或 以上使用阿卡波糖、二甲雙胍、噻唑烷二酮(TZD)、胰島素或磺酰脲/格列奈者，分別分類為阿卡波糖使用者、二甲雙胍使用者、TZD使用者、胰島素使用者或磺酰脲/格列奈使用者。根據中位年度累積劑量再將阿卡波糖使用者分類為高劑量使用者以及低劑量使用者。 Based on the NHIRD prescription record, there are 28 days in a year before the indicator date or The above uses acarbose, metformin, thiazolidinedione (TZD), insulin or sulfonylurea/glinide, respectively classified as acarbose users, metformin users, TZD users, insulin users or Sulfonylurea/Glinide user. Acarbose users are classified as high-dose users and low-dose users based on the median annual cumulative dose.

[潛在的干擾因素] [potential interference factors]

地理區域、DM持續時間以及DM終末段器官疾病(ICD9-CM 代碼250.1-9)列為潛在干擾因素。選擇地理區域是因為在以前的研究中發現其語RA風險相關。另外，DM持續時間以及DM終末段器官疾病則代表DM的嚴重度。DM持續時間定義為初始抗糖尿病藥之日至指標日期的期間。其他的干擾因素包含Deyo等人改寫之察爾森共病症指標(Charlson comorbidity index,CCI)(Deyo RA,Cherkin DC,Ciol MA.Adapting a clinical comorbidity index for use with ICD-9-CM administrative databases.J Clin Epidemiol.1992；45(6)：613-619)、如已往研究中定義的牙周炎(Chen HH,HuangN,Chen YM,et al.Association between a history of periodontitis and the risk of rheumatoid arthritis：a nationwide,populatior-based,case-control study.Ann Rheum Dis.2013；72(7)：1206-1211)，以及在指標日一年前使用史他汀(stain)藥物28天。 Geographical areas, DM duration, and DM terminal organ disease (ICD9-CM code 250.1-9) are listed as potential interference factors. The geographical area was chosen because it was found in the previous study that the RA risk was related. In addition, DM duration and DM terminal organ disease represent the severity of DM. The DM duration is defined as the period from the date of the initial antidiabetic drug to the date of the indicator. Other interfering factors include the Charlson comorbidity index (CCI) rewritten by Deyo et al. (Deyo RA, Cherkin DC, Ciol MA. Adapting a clinical comorbidity index for use with ICD-9-CM administrative databases.J Clin Epidemiol. 1992; 45(6): 613-619), periodontitis as defined in previous studies (Chen HH, Huang N, Chen YM, et al. Association between a history of periodontitis and the risk of rheumatoid arthritis:a Nationwide, populatior-based, case-control study. Ann Rheum Dis. 2013; 72(7): 1206-1211), and the use of statin drugs one year before the index day 28 days.

[結果] [result]

由以上的統計結果，共有723位RA病患(年齡16歲)作為RA 案例，以及7,230位匹配的分RA病患作為非-RA控制組(參下表1)。女性與男性的比例為2.5：1，案例之平均年齡(±SD)為61.7(±10.8)歲，控制組則為61.8(±10.5)歲。下表2則比較了RA案例與非-RA控制組的臨床特點。使用二甲雙胍、噻唑烷二酮(TZD)、胰島素及磺酰脲/格列奈的案例的比例高於控制組的比例。 From the above statistics, there are 723 RA patients (age 16 years old) as a RA case, and 7,230 matched RA patients as non-RA control group (see Table 1 below). The ratio of female to male was 2.5:1, the average age of the case (±SD) was 61.7 (±10.8) years, and the control group was 61.8 (±10.5) years old. Table 2 below compares the clinical features of the RA case with the non-RA control group. The proportion of cases using metformin, thiazolidinedione (TZD), insulin and sulfonylurea/glinide was higher than that of the control group.

在下表3中，調整了潛在的干擾因素後，每年投予超過16,950 mg的阿卡波糖使用者相較於非使用者，其RA風險較低(OR,0.60；95% CI,0.41-0.89)；胰島素的使用則為RA風險的另一個保護因子(OR,0.62；95% CI,0.45-0.87)；RA發展的顯著風險因子包含使用二甲雙胍(OR,1.26；95% CI,1.06-1.49)、較長的DM持續時間(每增加一年，OR,1.22；95% CI,1.18-1.27)、DM終末段器官疾病的發生(OR,2.67；95% CI,2.23-3.19)以及CCI(OR,1.281；95% CI,1.26-1.31)。 In Table 3 below, after adjusting for potential interference factors, more than 16,950 is administered annually. The acarbose user of mg had a lower risk of RA compared with non-users (OR, 0.60; 95% CI, 0.41-0.89); insulin use was another protective factor for RA risk (OR, 0.62) 95% CI, 0.45-0.87); significant risk factors for RA development include the use of metformin (OR, 1.26; 95% CI, 1.06-1.49), longer DM duration (every additional year, OR, 1.22; 95) % CI, 1.18-1.27), DM terminal organ disease (OR, 2.67; 95% CI, 2.23-3.19) and CCI (OR, 1.281; 95% CI, 1.26-1.31).

由以上可以發現，DM病患若每年投予16,950mg以上的阿卡 波糖，將可降低罹患RA的風險。 From the above, it can be found that DM patients who administer more than 16,950 mg of Aka per year Wave sugar will reduce the risk of RA.

實施例2-小鼠實驗確認阿卡波糖治療類風濕性關節炎Example 2 - Mouse experiment confirmed acarbose treatment of rheumatoid arthritis

[膠原蛋白誘發小鼠關節炎並以阿卡波糖治療] [Collagen induced arthritis in mice and treated with acarbose]

依據習知方法(Inglis JJ,Simelyte E,McCann FE,Criado G, Williams RO.Protocol for the induction of arthritis in C57BL/6 mice.Nat Protoc.2008；3(4)：612-618)對DBA/1(6至8週)小鼠進行膠原蛋白誘發關節炎(collagen-induced arthritis CIA)。將阿卡波糖(DXM，Sigma-Aldrich公司)溶解並以水稀釋；小鼠隨機分成三組，分別給予500mg/kg/day劑量的阿卡波糖、100mg/kg/day劑量的阿卡波糖，以及水作為對照組，在第一次免疫後的7至38天每天投予兩次；小鼠腳掌關節炎嚴重程度的臨床表徵以習知的評分系統(Zhou R,Tang W,Ren YX,et al.(5R)-5-hydroxytriptolide attenuated collagen-induced arthritis in DBA/1 mice via suppressing interferon-gamma production and its related signaling.J Pharm Exp Ther.2006；318(1)：35-44)，以雙盲方式進行評價。 According to the conventional method (Inglis JJ, Simelyte E, McCann FE, Criado G, Williams RO. Protocol for the induction of arthritis in C57BL/6 mice. Nat Protoc. 2008; 3 (4): 612-618) on DBA/1 (6 to 8 weeks) mice were subjected to collagen-induced arthritis (CIA). Acarbose (DXM, Sigma-Aldrich) was dissolved and diluted with water; mice were randomly divided into three groups, each given a dose of 500 mg/kg/day of acarbose and a dose of 100 mg/kg/day of Akapo. Sugar, as well as water as a control group, were administered twice daily for 7 to 38 days after the first immunization; the clinical characterization of the severity of mouse foot arthritis was performed using a conventional scoring system (Zhou R, Tang W, Ren YX). , et al. (5R)-5-hydroxytriptolide attenuated collagen-induced arthritis in DBA/1 mice via suppressing interferon-gamma production and its related signaling. J Pharm Exp Ther. 2006; 318 (1): 35-44), Evaluation in a double-blind manner.

[組織病理學分析] [histopathological analysis]

從每組中各一共三隻小鼠在第38天取得代表性的踝關節，於 石蠟中進行固定、脫鈣及包埋。各關節切片(5μM)後以蘇木紫及伊紅染色(H&E)，接著跟據習知的方法(Wooley PH.Collagen-induced arthritis in the mouse.Methods Enzymol.1988；162：361-373.)以顯微鏡觀察。滑膜增生、細胞浸潤、軟骨破壞以及骨侵蝕的組織病理變化，由病理學家進行盲目分級，並根據以下標準給予0~3分，0：沒有變化；1：輕度變化；2：中度變化；3：重度變化。 A total of three mice from each group obtained representative ankle joints on day 38 and fixed, decalcified and embedded in paraffin. Each joint section (5 μM) was stained with hematoxylin and eosin (H&E) followed by a conventional method (Wooley PH. Collagen-induced arthritis in the mouse. Methods Enzymol. 1988; 162:361-373.) Observed by a microscope. Synovial hyperplasia, cell infiltration, cartilage destruction, and histopathological changes of bone erosion were blindly graded by pathologists and given 0 to 3 points according to the following criteria, 0: no change; 1: mild change; 2: moderate Change; 3: Severe change.

[測定細胞激素含量] [Measurement of cytokine content]

各組在第38天收集小鼠腳掌組織，並在液態氮中冷凍，揭著 樣品以含有蛋白酶抑制劑中的PBS，以組織整塑器均質化。將均漿離心30分鐘，收集上清液並稀釋至每毫升1mg/ml組織蛋白做為原液。TNF-a、IL-1b、IL-6、IFN-g、IL-17、IL-10及TGF-b(R&D Systems)的含量以ELISA套組(Peprotech)測定。 Each group collected mouse paw tissue on day 38 and frozen in liquid nitrogen, revealing The sample was homogenized in a tissue homogenizer with PBS in a protease inhibitor. The homogenate was centrifuged for 30 minutes, and the supernatant was collected and diluted to 1 mg/ml tissue protein per ml as a stock solution. The contents of TNF-a, IL-1b, IL-6, IFN-g, IL-17, IL-10 and TGF-b (R&D Systems) were determined in an ELISA kit (Peprotech).

[抗膠原第II型IgG抗體產生的分析] [Analysis of anti-collagen type II IgG antibody production]

小鼠在第CII免疫接種後的第38天抽血，其血清利用抗CII IgG抗體以ELISA滴定分析。簡言之，將ELISA微孔盤(Thermo Fisher Scientific,NY,USA)的各孔塗覆10μg/200ml之第II型雞膠原並於4℃下培育過夜。為避免非特異性的結合，將抗體清洗並以3%牛血清蛋白(BSA)(BD Biosciences)的Tris緩衝液阻斷；受測血清進行系列稀釋，並加入孔盤中4℃下培育過夜。經過5至7次的清洗後，結合的IgG透過與HRP-共軛的綿羊抗小鼠IgG以1：5000稀釋做為二抗來偵測；清洗後，孔盤以ABTS(Roche Diagnostic Systems,IN,USA)做為基質反應，並以H₂SO₄停止反應，以ELISA讀數器在 450nm處量測吸光值。 Mice were bled on day 38 after CII immunization and their serum was analyzed by ELISA titration using anti-CII IgG antibodies. Briefly, each well of an ELISA microplate (Thermo Fisher Scientific, NY, USA) was coated with 10 μg/200 ml of type II chicken collagen and incubated overnight at 4 °C. To avoid non-specific binding, the antibodies were washed and blocked with 3% bovine serum albumin (BSA) (BD Biosciences) in Tris buffer; the test sera were serially diluted and incubated overnight at 4 °C in a well plate. After 5 to 7 washes, the bound IgG was detected by diluting 1:5000 with HRP-conjugated sheep anti-mouse IgG as a secondary antibody; after washing, the well plate was ABTS (Roche Diagnostic Systems, IN) , USA) As a matrix reaction, the reaction was stopped with H ₂ SO ₄ and the absorbance was measured at 450 nm using an ELISA reader.

[細胞增生] [cell hyperplasia]

免疫後第38天，取CIA小鼠的腹股溝淋巴結，以40μg/ml全 變性基CII培養3天。在終止培養的18個小時前，將1μCi之[³H]]胸苷脈衝至各培養盤中，透過Matrix 96直接電離貝他計數器(Packard Instrument,Meridian,CT)，以每分鐘計次(counts per minute,cpm)測定細胞增生。 On the 38th day after immunization, the inguinal lymph nodes of CIA mice were taken and cultured for 3 days at 40 μg/ml of fully denatured CII. 1 μCi of [ ³ H] thymidine was pulsed into each plate 18 hours before the termination of culture, and counted per minute by Matrix 96 direct ionization beta counter (Packard Instrument, Meridian, CT). Per minute, cpm) Cell proliferation was measured.

[結果] [result]

CIA已廣泛地應用於引起展現出人類RA病理特徵的關節 炎。首先調查是否阿卡波糖可阻止CIA於DBA/1小鼠模型中的進展。如圖1所示，給予水的CIA小鼠(對照組)在第24天有嚴重的CIA。然而，相較於對照組，以500mg/kg/day阿卡波糖治療的小鼠則在CIA發生率(圖1A)及患病腳掌數量(圖1B)有顯著地減少，且目視確認下有較不嚴重的腫脹、紅斑以及後腳掌的關節僵硬(圖1C)和關節炎評分(圖1D)；以阿卡波糖100mg/kg/day口服治療則在CIA的發展及嚴重程度沒有顯著的效果。 CIA has been widely used to cause joints that exhibit the pathological features of human RA. inflammation. First investigate whether acarbose can prevent progression of CIA in the DBA/1 mouse model. As shown in Figure 1, water-administered CIA mice (control group) had severe CIA on day 24. However, compared with the control group, mice treated with 500 mg/kg/day acarbose had a significant reduction in the incidence of CIA (Fig. 1A) and the number of affected feet (Fig. 1B), and visually confirmed Less severe swelling, erythema, and joint stiffness in the hind paw (Figure 1C) and arthritis score (Figure 1D); oral treatment with acarbose 100 mg/kg/day had no significant effect on the development and severity of CIA .

為了進一步評估關節炎，小鼠踝關節的組織學(H&E)研究在 實驗結束時進行。與觀察到的減緩的腳掌腫脹及臨床評分一致，投予500mg/kg/day阿卡波糖的小鼠在以下有顯著的降低：發炎細胞的浸潤、滑膜增生以及腳踝關節的骨侵蝕(圖1E-F)。接著以ELISA測定腳掌組織及血清中選擇性細胞激素的含量，如圖2A及圖2B所示，阿卡波糖治療的小鼠與對照CIA小鼠相比，其腳掌組織以及血清中的發炎細胞激素如TNF-alpha、IL-6及IL-17明顯地減少。這些結果顯示，以口服阿卡波糖可有效地預防CIA的發展以及抑制關節的局部炎症。 To further assess arthritis, histological (H&E) studies of the ankle joint in mice At the end of the experiment. Consistent with the observed slowed paw swelling and clinical score, mice given 500 mg/kg/day acarbose had a significant reduction in infiltration of inflamed cells, synovial hyperplasia, and bone erosion of the ankle joint (figure 1E-F). Next, the content of selective cytokines in the paw tissue and serum was measured by ELISA. As shown in FIG. 2A and FIG. 2B, the acarbose-treated mice had ampules in the soles of the feet and serum in comparison with the control CIA mice. Hormones such as TNF-alpha, IL-6 and IL-17 are significantly reduced. These results show that oral acarbose can effectively prevent the development of CIA and inhibit local inflammation of joints.

在CIA模型中，抗體以及T細胞反應在以第II型膠原免疫後的 疾病病因有所相關。因此，進一步量測血清中的C-II特異性抗體以及淋巴結中的CII特異性T淋巴球擴展。如圖3A所示，使用500mg/kg/day阿卡波糖治療導致抗CII IgG產生的顯著下降，再者，與對照組CIA小鼠獲得的細胞相比，由阿卡波糖治療的小鼠中取得的淋巴結細胞顯示CII抗原刺激增生的顯著降低(圖3B)，且膠原刺激的淋巴結細胞之IL-17以及IFN-γ產生降低。相反地，IL-10則在阿卡波糖治療的CIA小鼠中有增加的表現量(圖3C)。這些數據顯示無論是體液或細胞的免疫反應均會受到阿卡波糖治療的抑制。 In the CIA model, antibodies and T cell responses are immunized with type II collagen. The cause of the disease is related. Therefore, C-II specific antibodies in serum and CII-specific T lymphocyte expansion in lymph nodes were further measured. As shown in Figure 3A, treatment with 500 mg/kg/day acarbose resulted in a significant decrease in anti-CII IgG production, again, mice treated with acarbose compared to cells obtained from control CIA mice. Lymph node cells obtained in the test showed a significant decrease in CII antigen-stimulated proliferation (Fig. 3B), and IL-17 and IFN-γ production in collagen-stimulated lymph node cells was decreased. In contrast, IL-10 showed an increased amount of expression in acarbose-treated CIA mice (Fig. 3C). These data show that both body fluids and cellular immune responses are inhibited by acarbose treatment.

實施例3-阿卡波糖與托法替尼個別及合併投予之CIA小鼠實驗比較Example 3 - Comparison of Acarbose and Tofacitini in Individual and Combined CIA Mice

[不同劑量托法替尼對CIA小鼠之功效評估] [Evaluation of the efficacy of different doses of tofartinib in CIA mice]

使用如實施例2之CIA小鼠，小鼠隨機分成四組，分別給予1mg/kg/day劑量、5mg/kg/day劑量、25mg/kg/day劑量的托法替尼以及水作為對照組，在第一次免疫後的7至38天每天投予兩次；小鼠腳掌關節炎嚴重程度的臨床表徵以習知的評分系統。 Using the CIA mice as in Example 2, the mice were randomly divided into four groups, and a dose of 1 mg/kg/day, a dose of 5 mg/kg/day, a dose of 25 mg/kg/day of tofacitinib, and water were used as a control group, respectively. Two doses were administered daily from 7 to 38 days after the first immunization; the clinical characterization of the severity of mouse paw arthritis was performed using a conventional scoring system.

[結果] [result]

如圖4A所示，使用習知治療類風濕性關節炎之托法替尼，劑量越高，其對CIA小鼠臨床的表徵治療效果越好。 As shown in Figure 4A, the use of conventional tofapine in the treatment of rheumatoid arthritis, the higher the dose, the better the clinical characterization of CIA mice.

[阿卡波糖500mg/kg/day合併托法替尼1mg/kg/day對CIA小鼠之功效評估] [Akapoose 500mg/kg/day combined with tofartinib 1mg/kg/day evaluation of the efficacy of CIA mice]

使用如實施例2之CIA小鼠，小鼠隨機分成四組，分別給予1mg/kg/day劑量的托法替尼、500mg/kg/day劑量的阿卡波糖、1mg/kg/day劑 量的托法替尼合併500mg/kg/day劑量的阿卡波糖，以及水作為對照組，在第一次免疫後的7至38天每天投予兩次；小鼠腳掌關節炎嚴重程度的臨床表徵以習知的評分系統。 Using CIA mice as in Example 2, mice were randomized into four groups, each receiving a dose of 1 mg/kg/day of tofacitinib, a dose of 500 mg/kg/day of acarbose, and a dose of 1 mg/kg/day. A dose of tofacitinib combined with a 500 mg/kg/day dose of acarbose, and water as a control group, administered twice daily for 7 to 38 days after the first immunization; Clinical characterization is based on a conventional scoring system.

[結果] [result]

如圖4B所示，以一般阿卡波糖用於治療第二型糖尿病劑 量，相較於一般用於治療類風濕性關節炎之托法替尼1mg/kg/day的劑量，有更佳的功效；且兩者合併使用後，效果也較單一投予為佳。 As shown in Figure 4B, general acarbose is used to treat type 2 diabetes agents. The amount is better than the dose of 1 mg/kg/day of tofartinib which is generally used for the treatment of rheumatoid arthritis; and the combined effect of the two is better than single administration.

[阿卡波糖500mg/kg/day合併托法替尼5mg/kg/day對CIA小鼠之功效評估] [Akapoose 500mg/kg/day combined with tofacitinib 5mg/kg/day evaluation of the efficacy of CIA mice]

[結果] [result]

如圖4C所示，阿卡波糖500mg/kg/day與托法替尼5mg/kg/day均能用於治療類風濕性關節炎；且兩者合併使用後，效果更為顯著。 As shown in Fig. 4C, acarbose 500 mg/kg/day and tofacitinib 5 mg/kg/day can be used for the treatment of rheumatoid arthritis; and when the two are combined, the effect is more remarkable.

實施例4-阿卡波糖投予以咪喹莫特誘發乾癬之小鼠實驗Example 4 - Acarbose administration of imiquimod-induced dryness in mice

[誘發乾癬小鼠及投藥方式] [Inducing dry mice and administration methods]

利用習知技術獲得乾癬小鼠，簡單描述，將小鼠背部的毛髮剔除後，每天在小鼠背部連續每天塗抹62.5mg市售的咪喹莫特(IMQ)軟膏((Aldara；3M Pharmaceuticals)共塗抹6天，第7天，將小鼠拍照並犧牲後，取出背部皮膚，進行切片分析。阿卡波糖是在開始塗抹咪喹莫特(IMQ)軟膏前一個禮拜開始每天口服餵食直到實驗結束(第6天)，劑量分別為500mg/kg或100mg/kg。臨床評分標準是參考clinical Psoriasis Area and Severity Index(PASI)，主要是根據其紅斑(erythema)、脫屑(scaling)分成5個等級評分。0代表沒有變化，1至4分中，分數越高代表紅斑及脫屑現象越嚴重。 The dried mice were obtained by conventional techniques. Briefly, after removing the hair on the back of the mouse, 62.5 mg of commercially available imiquimod (IMQ) ointment ((Aldara; 3M Pharmaceuticals) was applied daily on the back of the mouse. After smearing for 6 days, on the 7th day, the mice were photographed and sacrificed, and the back skin was taken out for section analysis. Acarbose was orally administered daily until the end of the experiment one week before the application of imiquimod (IMQ) ointment. (Day 6), the dose is 500mg/kg or 100mg/kg respectively. The clinical scoring standard refers to the clinical Psoriasis Area and Severity Index (PASI), which is mainly divided into 5 grades according to its erythema and scaling. Score. 0 means no change, 1 to 4 points, the higher the score, the more serious the erythema and desquamation.

[結果] [result]

由圖5A及圖5B中可見，以咪喹莫特(IMQ)62.5mg/kg/day誘發 乾癬之小鼠在投予阿卡波糖500mg/kg/day後，治療效果顯著。 As can be seen from Fig. 5A and Fig. 5B, induced by imiquimod (IMQ) 62.5 mg/kg/day The dried mice were treated with acarbose 500 mg/kg/day, and the treatment effect was remarkable.

[組織病理學分析] [histopathological analysis]

從每組中各一隻小鼠在第7天取得之背部皮膚組織，於石蠟 中進行固定、脫鈣及包埋。切片(5μM)後以蘇木紫及伊紅染色(H&E)以顯微鏡觀察。如圖6所示。根據切片的結果，可以發現阿卡波糖500mg/kg可顯著減緩咪喹莫特(IMQ)誘導之小鼠，而造成之表皮肥厚(棘細胞増殖(acanthosis))、角質層增厚，並有殘存的細胞核，及有角化不良(parakeratosis)現象。 The skin tissue of the back obtained from the day 7 of each mouse in each group, in paraffin Fixation, decalcification and embedding. After sectioning (5 μM), staining with hematoxylin and eosin (H&E) was observed under a microscope. As shown in Figure 6. According to the results of the sectioning, it can be found that acarbose 500mg/kg can significantly slow down the imiquimod (IMQ)-induced mice, resulting in epidermal hypertrophy (acanthosis), thickening of the stratum corneum, and Remaining nuclei and parakeratosis.

綜合上開實驗可知，阿卡波糖確可用於治療類風濕性關節 炎、乾癬等免疫發炎病症，且證實其可調結IL-17的功效，因此，可更廣泛的用於與IL-17相關的免疫發炎病症。 Comprehensive open experiment shows that acarbose can be used to treat rheumatoid joints. Immune inflammatory conditions such as inflammation and dryness, and confirmed that it can regulate the efficacy of IL-17, and therefore, can be more widely used for immunoinflammatory diseases associated with IL-17.

Claims (7)

一種阿卡波糖於製備治療與IL-17相關之自體免疫發炎病症之藥物之用途。 Use of acarbose for the preparation of a medicament for treating an autoimmune inflammatory disorder associated with IL-17. 如請求項1之用途，其中該免疫發炎病症為：類風濕性關節炎、乾癬、第一型糖尿病、成人性史底耳氏症、視神經脊髓炎、克隆氏症、潰瘍性大腸炎、氣喘、慢性阻塞性肺病、風濕性多肌痛症、巨細胞動脈炎、全身性紅斑狼瘡、異位性皮膚炎、多發性硬化症、重症肌無力、牛皮癬、僵直性脊椎炎、肝硬化或乾癬性關節炎。 The use of claim 1, wherein the immune inflammatory condition is: rheumatoid arthritis, cognac, type 1 diabetes, adult sexual history, optic neuromyelitis, Crohn's disease, ulcerative colitis, asthma, Chronic obstructive pulmonary disease, rheumatic polymyalgia, giant cell arteritis, systemic lupus erythematosus, atopic dermatitis, multiple sclerosis, myasthenia gravis, psoriasis, ankylosing spondylitis, cirrhosis or dry joints inflammation. 如請求項1之用途，其進一步包含一第二藥物。 The use of claim 1 further comprising a second drug. 如請求項3之用途，其中該第二藥物選自由非類固醇抗發炎藥(NSAID)、COX-2抑制劑、生物分子、免疫調節劑、小分子免疫調節劑、疾病修飾抗風濕藥(DMARD)、黃嘌呤、抗膽鹼化合物、β受體協同劑、支氣管擴張劑、非類固醇免疫親和蛋白依存免疫抑制劑、維生素D類似物、補骨脂素、維生素A酸及5-胺基水楊酸所構成之群。 The use of claim 3, wherein the second drug is selected from the group consisting of a non-steroidal anti-inflammatory drug (NSAID), a COX-2 inhibitor, a biomolecule, an immunomodulator, a small molecule immunomodulator, and a disease modifying antirheumatic drug (DMARD). , Astragalus, anticholinergic compounds, beta-receptor synergists, bronchodilators, non-steroidal immunoaffinity protein-dependent immunosuppressants, vitamin D analogues, psoralen, vitamin A and 5-aminosalicylic acid The group formed. 如請求項4之用途，其中該疾病修飾抗風濕藥(DMARD)為托法替尼(tofactinib)。 The use of claim 4, wherein the disease modifying antirheumatic drug (DMARD) is tofactinib. 如請求項1至5項任一項之用途，其中該藥物係呈錠劑或膠囊之型式。 The use of any one of claims 1 to 5, wherein the medicament is in the form of a lozenge or capsule. 如請求項6之用途，其中該藥物為局部投予用藥或全身投予用藥。 The use of claim 6, wherein the drug is administered topically or systemically.